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Volume 4 Issue 3

International Pharmaceutical Industry

Supporting the industry through communication

Peer reviewed

Lessons Learned In Developing Cardiovascular and Metabolic Drugs Health Made in Germany Bag-on-Valve Optimal solution for pharma products Answering pharma’s need for outsourcing process ‘intensification’ technology For the future of biopharmaceutical production Lessons

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PUBLISHER: Mark A. Barker EDITORIAL MANAGER Jaypreet Dhillon BOOK MANAGER: Anthony Stewart BUSINESS DEVELOPMENT: Nigel Schneider DESIGN DIRECTOR: Ricky Elizabeth CIRCULATION MANAGER: Cecilia Stroe FINANCE DEPARTMENT: Martin Wright RESEARCH & CIRCULATION: Jean Baptiste Marty COVER IMAGE: iStockphoto © PRINTED BY: SW TWO UK PUBLISHED BY: Pharma Publications Unit J413, The Biscuit Factory Tower Bridge Business Complex 100 Clements Road, London SE16 4DG Tel: +44 (0)20 7237 2036 Fax: +44 (0)01 480 247 5316 Email: All rights reserved. No part of this publication may be reproduced, duplicated, stored in any retrieval system or transmitted in any form by any means without prior written permission of the Publishers. The next issue of IPI will be published in November 2012. ISSN No. International Pharmaceutical Industry ISSN 1755-4578. The opinions and views expressed by the authors in this magazine are not necessarily those of the Editor or the Publisher. Please note that although care is taken in preparation of this publication, the Editor and the Publisher are not responsible for opinions, views and inaccuracies in the articles. Great care is taken with regards to artwork supplied, the Publisher cannot be held responsible for any loss or damage incurred. This publication is protected by copyright. 2012 PHARMA PUBLICATIONS Volume 4 issue 3 Summer 2012


DIRECTORS: Martin Wright Mark A. Barker

Contents 06 Publisher’s letter Special Feature

International Pharmaceutical Industry

Supporting the industry through communication


Supporting the industry through communication


International Pharmaceutical Industry



08 Product Liability: Navigating a Restrictive Market Insurers are facing a growing number of product liability claims in what is becoming an increasingly litigious environment in the UK and Europe. Stricter regulation, a high-pressured economic environment and complex supply chains are only some of the factors driving this trend. As a result, it is often difficult to maintain access to broad and affordable product liability cover. Andrew Catton of Miller Insurance Services LLP explains why product liability insurance is becoming more restrictive for many pharmaceutical, biotech and medical equipment companies, and what organisations can do to maintain affordable cover. 10 Lessons Learned in Developing Cardiovascular and Metabolic Drugs: Introduction to the Series Paul Strumph, Rick Turner and Associates at Quintiles introduce a new Commentary Series entitled Lessons Learned in Developing Cardiovascular and Metabolic Drugs, that will appear in each issue of this journal. The goal of these papers is to provide readers with pragmatic information and insights that will facilitate well-informed decisions at critical “forks in the road” during preapproval clinical development programmes for cardiovascular and metabolic drugs. The commentaries will focus on specific challenges in clinical development, prioritised scenarios developed to succeed, outcomes, and lessons learned. Regulatory & Marketplace 16 Austrian Production Technologies - Better, faster, cheaper Austria has a wide variety of companies providing enabling technologies across the whole value chain of production processes. These range from good manufacturing practice knowhow for setting up pharmaceutical plants, to companies producing plasma, special enzymes and other biological substrates, and even stem cell products for the repair of bones, cartilage or muscular tissue. Sonja Polan at Life Science Austria discusses how service providers such as good clinical practice specialists help pharma or biotech companies and clinical trial units in Austria. 24 Extending Your Patent on a New Application for an Old Drug The period that is necessary to obtain a marketing authorisation to bring a new drug onto the market in the European Union significantly reduces the effective patent protection of that drug and, hence, the possibility to recover the high research and development costs. Stijn Lagaert at LC Patents provides an overview of how to compensate for that loss; the European Union member states provide supplementary protection certificates, which may extend the patent life of certain human and veterinary pharmaceuticals for up to five years. 30 Health – Made in Germany The market for healthcare is gaining steadily in global importance and offers great promise for innovation, growth and employment. Demographic developments, increasing health awareness, and even new products and technologies themselves, are driving the dynamics of further growth. Marion Luekemann at Germany Trade & Invest discusses how the worldwide market for healthcare holds great potential for further international co-operation, including with Germany’s healthcare industry.


Drug Discovery/Development & Delivery 38 Novel Factories for Novel Vaccines Vaccination is considered one of the most successful strategies for fighting infectious diseases, an area where there still remain significant unmet medical needs. It has been in the past difficult for business to find opportunities in this area, due to the long development times and to the risks involved in testing. Sancha Salgueiro, Charlotte Dyring and Wian de Jongh at ExpreS2ion Biotechnologies look into how a relief in this status has been observed recently. 44 Reduce Costs and Gain Expertise by Outsourcing your Cell Culture and Cryostorage Cell culture has evolved over more than a century into a core technique that is central to a diverse range of research and clinical applications. These range from the study of basic biological processes through to stem cell therapy and tissue engineering. As well as contributing to ethical progress by helping to reduce the use of animals, cell culture provides a number of key scientific benefits. Patrick Jackson at Vindon Scientific and Dr Carol Barker at XCellR8 provide an insight into how in pharmaceutical research and development, cell culture has an increasing number of cutting-edge roles, including a contribution to reduced attrition rates by allowing sophisticated early-stage safety and efficacy screening. 50 Straightforward or Challenging? Why It Matters Where You Place DM&PK Studies Medical research has been a major underlying factor leading to the dramatic reduction in overall morbidity and mortality in the 20th century. In his book, The Devil Under the Microscope, Thomas Hagar describes the discovery of sulfa drugs and the first observations of the effects of Penicillium fungi on bacteria. Jim Vrbanac at MPI Research discusses the science of drugs, pharmacology, and how this is divided into two distinct, separate but interactive domains: dynamics and kinetics. Labs & Logistics 56 Near-Infrared Fluorescent Methods for Protein Analysis Fluorescent dyes are important labelling tools for life science research. In recent years, a variety of classic protein analysis techniques have been updated with near-infrared fluorescence. Amy Schutz-Geschwender at LI-COR® Biosciences looks into how detection in the NIR spectrum (~700-900 nm) offers enhanced sensitivity and signal-to-noise ratios and why nearinfrared fluorescence is a powerful tool for protein analysis. Clinical & Medical Research 60 Paediatric Clinical Trials in Europe and the US: Challenges and Solutions Pharmaceutical and biotech companies have a regulatory obligation to prepare plans for and conduct studies in paediatric patient populations in both EU countries and the US. Colin Hayward at Premier Research highlights differences and similarities in the paediatric regulatory compliance strategies and priorities of companies in those regions. 64 Allergy and Biopharmaceutical Therapy, Part 1: Background and Current Treatment Regimens A fairly good knowledge base concerning various steps in allergic reactions now exists, but despite this knowledge the prevalence of allergic diseases is still increasing: in some areas of the industrialised world up to 50% of the population is affected. Juan Gispert at Quintiles provides the first paper in a two part series, describing the pathological role of immunoglobulin E (IgE) in allergic reactions, and how identification of this role has allowed physicians to treat reactions by addressing the underlying immunological mechanisms. The second paper in the series will discuss unique challenges for clinical trials in this therapeutic area. 2 INTERNATIONAL PHARMACEUTICAL INDUSTRY

68 Centralised ECGs Help Ensure Cardiac Safety of Obesity Drugs Obesity rates have dramatically increased over the past 20 years, making it one of the most prevalent health problems in the world. Despite this, there are only a very small number of adequately safe and effective obesity drugs available on the market to treat the condition. Robert Kleiman at ERT discusses how centralised ECG trials have emerged as a viable method for helping drug companies provide reliable cardiovascular data in their attempts to achieve regulatory compliance, while also benefiting from significantly improved data accuracy and reliability, time and cost savings, and access to breakthrough technologies. 72 Clinically-relevant in vitro and in vivo Oncology Models for Drug Discovery The success rate in predicting clinical efficacy of anti-cancer modalities using current xenograft models has been reported to be only 30-40%. Standard xenograft models use cell lines that are maintained in plastic and have adapted to grow independently of the tumour microenvironment, resulting in models with genetic and phenotypic characteristics distinct from that seen in the clinic. Rajendra Kumari and Martin Page at PRECOS look at why, in the attempt to reduce drug attrition and improve clinical predictivity, patient-derived xenograft tumours are being used to improve and refine preclinical modelling. 78 Action Duchenne – Skip Duchenne Campaign Nick Catlin at Charity Action Duchenne provides an overview of the research into a cure and effective medicines for Duchenne/ Becker Muscular Dystrophy. The charity, which is led by Duchenne families, aims to promote awareness of the condition, to improve care services, and provide access to a range of educational and support/development programmes for people living with Duchenne at every stage of the condition. This is achieved by working in partnership with government agencies, NHS and care organisations, other charities, academic, scientific and research groups, and biotech/pharmaceutical/drug discovery companies worldwide. Labs & Logistics 82 Reverse Osmosis for Laboratory Use Poor specification and maintenance can significantly reduce the quality of output from reverse osmosis water purification systems. Mark Bosley at Purite describes, in contrast, how a well-specified, well-managed RO water purification system from a trusted and experienced supplier can deliver to the laboratory an efficient, economical and reliable supply to an exceptionally high level of purity. Manufacturing 88 Answering Pharma’s Need for Outsourcing to Process ‘Intensification’ Technology for the Future of Biopharmaceutical Production. The chemical and pharmaceutical industry will face challenges in the future related to manufacturing costs and lowering environmental impact. Chemical and biopharmaceutical manufacturing need to be modernised. This will bring lower raw material usage, higher yields with fewer resources, more focused manufacturing processes, and reduced manufacturing cost as final results. Fritjof Linz at DSM Biologics discusses how application of technologies brings down its manufacturing costs through process intensification to offer shorter and more efficient scale-up of (bio)pharmaceuticals, speeding up development, better managing manufacturing volumes and driving down total costs.

Summer 2012 Volume 4 Issue 3

92 The Healthy Printer The printing of prescription, identification and barcode labels takes place every day in every pharmacy throughout the country. However, busy pharmacists probably never give a second thought to the printer they use, until it goes wrong, or unless it regularly causes problems such as jammed media and software incompatibility. Jörk Schüßler at Citizen Systems Europe explains why the increasingly powerful levels of functionality and added resilience offered by the latest label, barcode, portable and point-of-sale printers can offer long, trouble-free life to serve busy pharmacies.

126 Interview Feature Planning and Performing Clinical Trials in Children and Adolescents Jaypreet Dhillon, Editorial Manager of Pharma Publications, Interviews Dr. Hermann Schulz, Chief Executive Officer INTERLAB Central Lab Services – Worldwide GmbH, Munich, Germany, on why paediatric trials are coming into focus now.

98 A Fluid Process The control of fluid flow – both gas and liquid – forms an essential part of many different types of medical equipment, from boiler control on steam sterilisers and reagent dispensing in in vitro diagnostic equipment, to bulk delivery of gases or precise gas delivery in products such as ventilators. A wave of new and modified flow technologies has helped to create effective solutions for manufacturers of medical devices. Sherea Lizaso at Gems Sensors & Controls discusses why the maximum benefit of these advances can only be gained by working alongside an expert provider of fluidic technology from the early design stages. Packaging 102 Simplifying the Lives of Patients Drug delivery devices currently range among the major pharmaceutical trends. Their optimisation is largely influenced by the pharmaceutical industry, physicians and patients. Dr Jérôme Freissmuth at Robert Bosch GmbH discusses how present developments of new devices are increasingly focused on patients’ needs. They give top priority to simplified handling, reduced pain, continuous documentation and facilitated adherence to medication intervals. 106 Due Diligence for Leachables and Extractables in a Modern GMP Environment. Nearly two decades have passed since extractables and leachables became a primary consideration for qualifying packaging intended for pharmaceutical use. The complexity of medicines and associated delivery systems continues to evolve, challenging approaches for identifying and quantifying leachables. Regulatory guidance for extractables and leachables exists, but only provides general recommendations for ensuring patient safety. Diane Paskiet, Laura Stubbs and Heike Kofler at West Pharmaceutical Services provide an overview of extractable and leachable science, and discuss approaches, applications and risk-based strategies. 114 Labelling and its Role in Pharmaceutical Packaging The ever-increasing legislative demands for patient information on pharmaceutical labelling often poses a packaging challenge to manufacturers and brand-owners. Patient compliance is also a major issue for the industry. Poorly worded, unclear or confusingly presented user-instructions can lead to ineffective patient compliance, adverse reactions, side-effects or at worst fatality. Steve Moore at Fix-a-Form International Ltd discusses why pharmaceutical labelling goes far beyond functional requirements and fulfilling legal obligations. 120 Bag-on-Valve – Optimal Solution for Pharma Products With tax-based healthcare service burdened by increasingly higher costs for pharmaceuticals and the ever-more stringent regulation governing the introduction of a new drug onto the market, there is a growing trend moving from prescription (Rx) to non-prescription, over-the-counter (OTC) products. Magnus Hedman at Aurena discusses why it has turned out that the Bag-on-Valve (BoV) may in several cases be a suitable technology for products to make this switch, as marketers are looking for modern, innovative packaging that can attract consumers.


Summer 2012 Volume 4 Issue 3

Publisher’s letter Welcome to the latest edition of IPI. The summer holidays are now over, and we are looking forward to the third quarter of 2012. We are all busy preparing for the multitude of exhibitions and conferences ahead. One of the biggest contract services events – ICSE/CPHI – this year is being held at Feria de Madrid, Spain. Be sure to mark your calendar for 9-11 October 2012. IPI also welcomes you to ScanBalt Forum 2012! The annual ScanBalt Forum enters its second decade on 20-23 November in Tampere, Finland. This year, the life science community on top of Europe will gather in Tampere Hall, Scandinavia’s largest congress and concert centre, to exchange ideas and discuss the current state and future prospects of ScanBalt BioRegion. Industry partnering in the form of inlicensing has come to the fore over the past decade. According to data from Campbell Alliance, in 1995 drugs based on deals accounted for just over 20% of revenue, but by 2010 that figure spiked to 50% of big pharma’s revenue coming from in-licensed deals. This is why events like BIO-Europe in Hamburg, Germany, November 12-14, 2012 at the CCH Congress Center Hamburg are vital to the industry. BIOEurope provides one of the largest and most dynamic platforms where high-level industry executives and dealmakers can identify and meet with numerous potential partners. In this issue of IPI we bring you again a large variety of highly pertinent topics which will guide you efficiently through

the ever-changing landscape of the modern life sciences industry. We begin on page 8, with the fantastic article by Andrew Catton of Miller Insurance, on Product Liability: Navigating a Restrictive Market. Paul Strumph et al. of Quintiles look (on page 10) into Lessons Learned in Developing Cardiovascular and Metabolic Drugs: Introduction to the Series. The Regulatory & Marketplace section in this issue concentrates on Austrian Production Technologies Better, Faster, and Cheaper, an article by Sonja Polan, the International Marketing Manager of LISA – Life Science Austria. Stijn Lagaert of LC Patents opens the door for our readers (on page 24) with an insight into Extending your Patent on a New Application for an Old Drug and the implications of the Neurim ruling. Germany was among the top five countries in the world in terms of market size; Marion Luekemann of GTAI looks at the health industry “Made in Germany” on page 30. The Drug Discovery, Development & Delivery section carries articles from ExpreS2ion Biotechnologies (on page 38), concentrating on novel factories for novel vaccines. On page 44, we move to reducing costs and gaining expertise by Outsourcing with regard to Cell Culture and Cryostorage, with an article by Patrick Jackson of Vindon and Dr Carol Barker of XCellR8. Jim Vrbanac of MPI Research (on page 50) answers the question: Straightforward or Challenging? Why It Matters Where You Place DM&PK Studies. Near-Infrared Fluorescent Methods for Protein Analysis

are examined by Amy Geschwender of Licor (on page 56). Technology for the Future of Biopharmaceutical Production by Dr Fritjof Linz of DSM. The Healthy Printer article (on page 92) is by Jörk Schüßler, the European Marketing Manager for Citizen Systems Europe. Schüßler explains why the increasingly powerful levels of functionality and added resilience offered by the latest label, barcode, portable and point-of-sale printers can offer long, trouble-free life to serve busy pharmacies. Sherea Lisazo of Gems-Sensors provide an insight (on page 98) into the control of fluid flow – both gas and liquid – as they form an essential part of many different types of medical equipment. The Packaging section (on page 102), starts with Simplifying the Lives of Patients by Dr Jérôme Freissmuth of Bosch, and continues (on page 106) with Due Diligence for Leachables and Extractables in a Modern GMP Environment, by Diane Paskiet, Laura Stubbs and Heike Kofler of West Pharma. Labelling and its Role in Pharmaceutical Packaging is an issue covered by Steve Moore of fix-a-form (on page 114). And we round up this issue of IPI with Bagon-Valve – Optimal Solution for Pharma Products, by Magnus Hedman of Aurena Laboratories AB. I hope you all enjoy reading this latest edition of IPI, and we look forward to meeting you all at ICSE/CPHI Madrid at our stand #10B01 in Hall 10.

Jeffrey Litwin, M.D., F.A.C.C. Executive Vice President and Chief Medical Officer of ERT

Rick Turner, Senior Scientific Director, Quintiles Cardiac Safety Services & Affiliate Clinical Associate Professor, University of Florida College of Pharmacy

Mark A. Barker Publisher

Editorial Advisory Board Bakhyt Sarymsakova, Head of Department of International Cooperation, National Research Center of MCH, Astana, Kazakhstan Catherine Lund, Vice Chairman, OnQ Consulting

Jeffrey W. Sherman, Chief Medical Officer and Senior Vice President, IDM Pharma

Deborah A. Komlos, Senior Medical & Regulatory Writer, Thomson Reuters

Jim James DeSantihas, Chief Executive Officer, PharmaVigilant

Robert Reekie, Snr. Executive Vice President Operations, Europe, Asia-Pacific at PharmaNet Development Group

Diana L. Anderson, Ph.D president and CEO of D. Anderson & Company

Mark Goldberg, Chief Operating Officer, PAREXEL International Corporation

Sanjiv Kanwar, Managing Director, Polaris BioPharma Consulting

Franz Buchholzer, Director Regulatory Operations worldwide, PharmaNet development Group

Maha Al-Farhan, Vice President, ClinArt International, Chair of the GCC Chapter of the ACRP

Stanley Tam, General Manager, Eurofins MEDINET (Singapore, Shanghai)

Francis Crawley. Executive Director of the Good Clinical Practice Alliance – Europe (GCPA) and a World Health Organization (WHO) Expert in ethics Georg Mathis Founder and Managing Director, Appletree AG Heinrich Klech, Professor of Medicine, CEO and Executive Vice President, Vienna School of Clinical Research 6 INTERNATIONAL PHARMACEUTICAL INDUSTRY

Nermeen Varawalla, President & CEO, ECCRO – The Pan Emerging Country Contract Research Organisation Patrice Hugo, Chief Scientific Officer, Clearstone Central Laboratories

Stefan Astrom, Founder and CEO of Astrom Research International HB Steve Heath, Head of EMEA Medidata Solutions, Inc T S Jaishankar, Managing Director, QUEST Life Sciences

Summer 2012 Volume 4 Issue 3


Product Liability: Navigating a Restrictive Market Insurers are facing a growing number of product liability claims in what is becoming an increasingly litigious environment in the UK and Europe. Stricter regulation, a high-pressured economic environment and complex supply chains are only some of the factors driving this trend. As a result, it is often difficult to maintain access to broad and affordable product liability cover. Miller’s life sciences expert, Andrew Catton, explains why product liability insurance is becoming more restrictive for many pharmaceutical, biotech and medical equipment companies, and what organisations can do to maintain affordable cover. Claims Inflation Within the healthcare sector, there has been a noticeable trend of claims inflation in recent years. One of the driving factors is the presence of new and tighter regulations, including the US Consumer Product Safety Improvement Act of 2008, the impact of which is now being felt, and the incoming EU Consumer Rights Directive & EU Regulation on Common Sales Law. These new laws have increased scrutiny in what was already a highly-regulated industry. While the US market has always been highly litigious, there are signs that the UK and Europe are catching up. Recent reform of the consumer code to include class actions, or collective redress, has seen the frequency of product liability claims rise sharply in these markets. At the same time, the economic environment has brought more pressure to bear on life sciences organisations. In today’s globalised world, connected by ever-more complex supply chains, suppliers are often located in low-cost jurisdictions where the same level of quality control or oversight may not be in place. This trend has seen an increasing number of product recalls in which goods supplied from Asia to US or European 8 INTERNATIONAL PHARMACEUTICAL INDUSTRY

manufacturers fail to stand up to national regulations. Nevertheless, organisations are forced to balance their ongoing cost pressures against their risk and compliance responsibilities. Contract negotiations can help ensure organisations are not overly exposed to risks in the supply chain. More pertinent to the healthcare sector are the growing claims related to implantable medical devices. Here recent high-profile cases, including likely claims from the PIP breast implant saga, have dented insurers’ profitability and in turn led to a tightening of cover for product recall across the sector as a whole. Advice to Insurance Buyers With the rising cost of claims, product liability is inevitably becoming more restrictive. This is true across the whole spectrum of life sciences organisations, but particularly for those that produce, import, or supply implantable devices. Many insurers are imposing more onerous conditions and exclusions, wanting to see their clients take high retentions and requiring more detail on risk management procedures and controls that are in place. To avoid being unfairly penalised, life sciences organisations need to work with the right insurance and broking partners to ensure they have full access to the market and are suitably rewarded for maintaining a better-than-average risk profile. Communication is essential and the insureds are encouraged to demonstrate all the steps that have been taken to improve and maintain the safety of their products and devices. Bringing new products to market is particularly fraught with difficulty in the current climate. Pharmaceutical, biotech and medical equipment companies are encouraged to detail every stage with their underwriters, so that they have a clear understanding

of the extent of clinical trials required before a new product is released. Even sensitive information should be shared with insurance and broking partners, and here non-disclosure agreements can be of help. By gaining an insight into the development procedures, study results and the quality and extent of clinical trials that have taken place, insurers will be more comfortable taking on the liability risk of new products coming to market. By partnering with the right brokers and insurers, building up a relationship and keeping communication channels open, life sciences organisations can ensure that they continue to receive cost-effective insurance, tailored to their risk profile. In the current environment, maintaining an open dialogue and taking steps to improve your risk profile can make a crucial difference. Andrew Catton has worked in the insurance industry since 1971, joining Miller, a leading independent insurance broker, in 1996. Specialising in pharmaceutical, medical and life science product liability, clinical trials medical malpractice, professional indemnity, biomedical errors and omissions, product recall and intellectual property insurance, Andrew has spoken at numerous biotechnology and clinical trials conferences and contributed to multiple publications. Email:

Summer 2012 Volume 4 Issue 3


Lessons Learned in Developing Cardiovascular and Metabolic Drugs: Introduction to the Series Introduction This is the inaugural paper in a new Commentary Series entitled “Lessons Learned in Developing Cardiovascular and Metabolic Drugs” that will appear in each issue of this journal. This particular commentary provides an introduction to the cardiovascular and metabolic diseases that will be addressed in subsequent commentaries. This strategy means that future commentaries can focus specifically on lessons learned during the development of new drugs for these diseases without having to take the space to discuss them. The paramount importance of developing new drugs for these diseases is captured in the statistics provided in this commentary concerning their prevalence worldwide. The goal of these papers is to provide readers with pragmatic information and insights that will facilitate well-informed decisions at critical “forks in the road” during preapproval clinical development programmes for cardiovascular and metabolic drugs. The commentaries will focus on specific challenges in clinical development, prioritised scenarios developed to succeed, outcomes, and lessons learned. It is intended that insights will be provided from many perspectives, drawing on the cumulative expertise and experience of thought leaders within contract research organisations (CROs), academic research organisations (AROs), biopharmaceutical companies, regulatory agencies, and key opinion leaders (KOLs). Type 2 Diabetes Mellitus Consider first diabetes. The International Diabetes Federation (IDF) recently observed that, in 2011, there were 366 million people with diabetes, a figure expected to rise to 552 million by 2030. 1 While these numbers are already extremely disconcerting, the authors also observed that “Most people with diabetes live in low- and middle10 INTERNATIONAL PHARMACEUTICAL INDUSTRY

income countries, and these countries will also see the greatest increase over the next 19 years.” As one country-specific example, data from the United States 2011 National Diabetes Fact Sheet 2 state that 25.8 million children and adults in the United States—8.3% of the population—have diabetes, with 18.8 million being diagnosed and 7.0 million people being undiagnosed. Another 79 million people have prediabetes. The importance of developing new drugs for type 2 diabetes mellitus (T2DM) lies in the fact that, even though there are successful drugs currently on the market, since insulin resistance increases as patients age it is likely that new drugs will need to be added to their regimens. As the European Medicines Agency (EMA) noted, glucose control in type 2 diabetes deteriorates progressively over time, and, after failure of diet and exercise alone, patients require on average a new intervention with glucose-lowering agents every three to four years to obtain/retain good control. 3 Glycohaemoglobin (HbA1c) is the most widely accepted measure of overall, long-term blood glucose control in patients with diabetes. It reflects the mean glucose concentration over the past 2-3 months and thus the immediate clinical consequences of diabetes (hyperglycaemia and its associated symptoms). When the results of an HbA1c laboratory test are used to diagnose diabetes the following classifications are defined: 4 • Normal: Less than 5.7% • Pre-diabetes: 5.7% to 6.4% • D iabetes: 6.5% or higher Traditionally, HbA1c has been used as a biomarker to evaluate longterm glycaemic control. Compelling data from the UK Prospective Diabetes Study (UKPDS) Group have demonstrated that lowering HbA1c in patients with T2DM reduces the risk of microvascular disease, 5 although robust evidence for a reduction in macrovascular disease

has not been demonstrated to date. The evidence that lowering HbA1c reduces microvascular disease argues for the continued use of this biomarker in clinical trials, and both the EMA and the US Food and Drug Administration (FDA) regard it as an appropriate primary endpoint to support a marketing claim based on glycaemic control. 3,6 However, EMA and FDA regulatory landscapes now also address macrovascular factors explicitly in the form of demonstrating that a drug does not unacceptably increase the risk of such disease, quantified by evaluation of the relative occurrence of major adverse cardiovascular events (MACE) endpoints (nonfatal myocardial infarction, nonfatal stroke, cardiovascular death, and possibly others such as urgent revascularisation and acute coronary syndrome) in the drug and control treatment groups (see Turner and Strumph 7 for a detailed review). The ramifications of these increased regulatory burdens will be discussed in a future commentary, along with suggestions for meeting the regulatory requirements as efficiently as possible. Obesity Consider next obesity, defined by the World Health Organization (WHO) as excessive fat accumulation that presents a risk to health. 8 Like diabetes, obesity is a major international public health concern. Recent evidence also supports obesity as an independent risk factor for cardiovascular disease, in particular for fatal coronary heart disease events. 9 While the total prevalence of obesity varies considerably by age, socio-economic status, culture, sex, race and other characteristics in each country, the WHO estimated that in 2005 at least 400 million people globally were obese, and projected that figure to increase to over 700 million people by 2015. 10 Just as HbA1c is used to define disease Summer 2012 Volume 4 Issue 3

SPECIAL FEATURE status in diabetes, the body mass index (BMI) classification system adopted by the US National Institutes of Health (NIH) 11 and WHO 12 defines the following:

the recommendation in this case. A future commentary will discuss this Committee Meeting and FDA’s decision in detail.

• Underweight — BMI <18.5 kg/m2 • Normal weight — BMI ≥18.5 to 24.9 kg/m2 • Overweight — BMI ≥ 25.0 to 29.9 kg/m 2 • Obesity — BMI ≥ 30 kg/m 2 Obesity Class I — BMI of 30.0 to 34.9 kg/m2 Obesity Class II — BMI of 35.0 to 39.9 kg/m2 Obesity Class III — BMI ≥40 kg/m2. This type of obesity is also referred to as severe, extreme, or morbid obesity.

Cardiovascular Disease The need for cardiovascular drugs is reflected by the fact that cardiovascular disease is the leading cause of death in many countries, including the United States, where it is responsible for 17% of US health expenditures. However, these costs are expected to rise substantially in the next two decades. By 2030, 40.5% of the population is projected to have some form of CVD. 18 Using methodology to project future costs of care for CVD that avoided ‘double counting’ of costs for patients with multiple cardiovascular conditions, the authors of a recent authoritative paper projected that between 2010 and 2030 the total direct real medical costs (calculated in 2008 US$) would triple from $273 billion to $818 billion: Real indirect costs due to lost productivity for all CVD were estimated to increase from $172 billion to $276 billion, an increase of 61%. 18 The development of some classes of cardiovascular drugs with very favourable benefit-risk profiles has been efficient, e.g., the threehydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, more commonly known as statins. These drugs are widely available, inexpensive, and a potent therapy for treating elevated cholesterol in both primary and secondary prevention. 19-21 Clinical development programmes for other drugs have been less successful, with benefit-risk considerations being a primary issue. Consider the example of torcetrapib. This agent was demonstrated to inhibit the development of atherosclerosis in non-clinical studies (a rabbit model), and, in early-phase clinical studies, to increase high-density lipoprotein cholesterol (HDLc) by 60% to 100% while at the same time lowering low-density lipoprotein cholesterol (LDLc, the “bad cholesterol”) by up to 20%. 22 This evidence suggested a cardioprotective effect of torcetrapib. The Investigation of Lipid Level Management to Understand its

Currently, about 34% of adults twenty years of age or older in the US and about 20% of European adults, on aggregate, are obese. 10,13 Because excess weight contributes to the development and aggravation of a large number of chronic diseases, including diabetes, hypertension, dyslipidemia, cardiovascular disease, and cancer of various organs, the economic burden (both direct and indirect costs) from the increasing prevalence of obesity is enormous. Various antiobesity drugs have been plagued with safety concerns or patient non-adherence because of unpleasant side-effects (see Caveney et al. 14 for a review). Therefore, the need for additional antiobesity drugs that are both safe and effective is considerable. The regulatory landscape for safety assessments in the US appears likely to become more stringent, however, following a meeting of the FDA’s Endocrinologic and Metabolic Drugs Advisory Committee held on March 28 th and 29 th 2012. 15,16 Committee members voted 17-6 that sponsors be required to prospectively exclude unacceptable cardiovascular risk even for new obesity drugs without a theoretical risk or signal for cardiovascular harm by employing a cardiovascular outcomes trial or an appropriately sized meta-analysis of MACE data from Phase II and Phase III trials. The FDA is not required to follow the recommendations of its advisory committees, but, given that such guidance would essentially follow their guidance for the prospective exclusion of unacceptable risk for drugs for T2DM, 17 it is reasonable to think that the FDA may follow 12 INTERNATIONAL PHARMACEUTICAL INDUSTRY

Impact in Atherosclerotic Events (ILLUMINATE) trial tested the proposition that torcetrapib would reduce the risk of clinical cardiovascular events. However, torcetrapib was associated with an increased risk of major cardiovascular events, and also increased mortality (from both cardiovascular and noncardiovascular causes). The drug’s sponsor terminated ILLUMINATE prematurely at the recommendation of the trial’s independent steering committee, based on advice from the Independent Data and Safety Monitoring Board. 23 Fortunately, development of other drugs in this class, i.e., cholesteryl ester transfer protein (CETP) inhibitors, is progressing more successfully to date. 24 Cardiodiabesity While the term cardiodiabesity has not yet become as common in the medical lexicon as other relatively new relevant terms, it meaningfully encapsulates all three diseases states discussed in this paper: Cardiovascular disease, diabetes, and obesity (see additional references 25,26 for further discussion). As Kovacic and Fuster 27 noted, “Obesity, insufficient physical exercise, diabetes, and advancing age are major risk factors for developing cardiovascular disease that are currently increasing in prevalence.” While these disease states and their treatment can be usefully discussed separately, ultimate discussions must take account of the comorbidities and the complex natures of pharmacotherapy for this triumvirate of conditions of clinical concern. Future commentaries in this series will do so. References 1. W hiting DR, Guariguata L, Weil C, Shaw J, IDF Diabetes Atlas: Global estimates of the prevalence of diabetes for 2011 and 2030. Diabetes Research and Clinical Practice, 94:311-321. 2. A merican Diabetes Association. http:// w w w. d i a b e t e s . o r g / d i a b e t e s - b a s i c s / diabetes-statistics (Accessed 27 March 2012) 3. E MA. Guideline on clinical investigation of medicinal products in the treatment Summer 2012 Volume 4 Issue 3

Regulatory & Market Place







8. 9.







of diabetes mellitus, draft. September 2011. en_GB/document_library/Scientific_ guideline/2011/10/WC500115945.pdf. M  edlinePlus Medical Encyclopedia. ency/article/003640.htm U  K Prospective Diabetes Study (UKPDS) Group. Intensive blood-glucose control with sulphonylureas or insulin compared with conventional treatment and risk of complications in patients with type 2 diabetes (UK¬PDS 33). Lancet. 1998:352:837-853. F  DA. Guidance for Industry. Diabetes mellitus: developing drugs and therapeutic biologics for treatment and prevention. February 2008. www. ComplianceRegulatoryInformation /Guidances/UCM071624.pdf. T  urner JR, Strumph P. FDA and EMA actions regarding the cardiovascular safety of drugs for type 2 diabetes mellitus, 2007-2012: An overview of respective regulatory landscapes. Journal for Clinical Studies, 2012;(3):2224. W  orld Health Organization. topics/obesity/en L ogue J, Murray HM, Welsh P, et al. Obesity is associated with fatal coronary heart disease independently of traditional risk factors and deprivation. Heart. 2011;97(7):564-568. F legal KM, Carroll MD, Ogden CL, Curtin LR. Prevalence and trends in obesity among US adults, 1999–2008. Journal of the American Medical Association. 2010; 303: 235–241. C linical guidelines on the identification, evaluation, and treatment of overweight and obesity in adults--The Evidence Report. National Institutes of Health. Obesity Research. 1998; (6 Suppl 2):51S-209S. W orld Health Organization. Obesity: preventing and managing the global epidemic. Report of a WHO convention, Geneva, 1999. WHO Technical Report Series 894, Geneva 2000. W orld Health Organization Europe. 10 things you need to know about obesity. November 2006. C aveney E, Caveney BJ, Somaratne R, Turner JR, Gourgiotis L, 2011, Pharmaceutical interventions for obesity: A public health perspective. Diabetes, Obesity, and Metabolism, 13:490-497. A genda for Advisory Committee Meeting.


w w w. f d a . g o v / d o w n l o a d s / A d v i s o r y Committees/CommitteesMeeting Materials/Drugs/Endocrinologic andMetabolicDrugsAdvisory Committee/UCM299125.pdf 16. Voting Questions for Advisory Committee Meeting. / downloads/AdvisoryCommittees/ CommitteesMeetingMaterials/ Drugs/Endocrinologicand M e t a b o l i c D r u g s A d v i s o r y Committee/UCM299126.pdf 17. FDA. Guidance for Industry. Diabetes Mellitus—Evaluating cardiovascular risk in new antidiabetic therapies to treat type 2 diabetes. December 2008. h t t p : / / w w w. f d a . g o v / d o w n l o a d s / Drugs/GuidanceCompliance RegulatoryInformation/Guidances/ ucm071627.pdf 18. H eidenreich PA, Trogdon JG, Khavjou OA, et al. Forecasting the future of cardiovascular disease in the United States: A policy statement from the American Heart Association. Circulation. 2011;123:933-944. 19. M inder CM, Blaha MJ, Horne A, et al. Evidence-based use of statins for primary prevention of cardiovascular disease. American Journal of Medicine. 2012 Mar 1. [Epub ahead of print] 20. Tonelli M, Lloyd A, Clement F, et al: Alberta Kidney Disease Network. Efficacy of statins for primary prevention in people at low cardiovascular risk: a metaanalysis. Canadian Medical Association Journal. 2011;183(16):E1189-1202. 21. L ewis SJ. Lipid-lowering therapy: who can benefit? Vascular Health and Risk Management. 2011;7:525-534. 22. R ader DJ. Editorial. Illuminating HDL — Is it still a viable therapeutic target? New England Journal of Medicine. 2007;357:2180-2183. 23. B arter PJ, Caulfield M, Eriksson M, et al. Effects of torcetrapib in patients at high risk for coronary events. New England Journal of Medicine. 2007;357:21092122. 24. G hosh RK, Ghosh SM. Current status of CETP inhibitors in the treatment of hyperlipidemia: An update. Current Clinical Pharmacology. March 20, 2012 [Epub ahead of print] 25. Turner JR, Strumph P, 2012, The moral imperative of improving patient adherence to pharmacotherapy for cardiodiabesity, Part I: A focus on type 2 diabetes mellitus. Journal for Patient

Compliance, 2(1):32-36. 26. R  ichards AL, Turner JR, 2012, The moral imperative of improving patient adherence to pharmacotherapy for cardiodiabesity, Part II: A focus on Cardiovascular Disease. Journal for Patient Compliance. 2012;2(2):36-41., in press. 27. K ovacic JC, Fuster V. From treating complex coronary artery disease to promoting cardiovascular health: Therapeutic transitions and challenges, 2010-2020. Clinical Pharmacology and Therapeutics. 2011;90(4):509-518.

Paul Strumph, MD, is Vice President, Therapeutic Strategy Lead, Cardiovascular and Metabolic Therapeutic Area, and Chief Medical Officer, North America Region, Quintiles. He is a diplomate of the American Boards of Internal Medicine, Pediatrics, Adult Endocrinology Diabetes and Metabolism, and Pediatric Endocrinology, and an author of more than a dozen articles in scientific journals and magazines, including Nature, Clinical Pediatrics, and the Journal of Clinical Endocrinology and Metabolism. Email: Erica Caveney, MD, Loukas Gourgiotis, MD, Adrienne L. Richards, MD, Hans van Dijk, and J. Rick Turner, PhD, is Senior Director, Clinical Communications, Quintiles. He is a clinical triallist, Editor-in-Chief of the Drug Information Journal, a Senior Fellow at the Center for Medicine in the Public Interest, and a Fellow of the Society for Behavioral Medicine. He is an author of more than 100 peer-reviewed papers and articles in professional journals, and has authored and edited 14 books. Email: All authors are members of Quintiles’ Cardiovascular and Metabolic Therapeutic Area Writing Committee.

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Austrian Production Technologies â&#x20AC;&#x201C; Better, Faster, Cheaper Austria has a wide variety of companies providing enabling technologies across the whole value chain of production processes. These range from good manufacturing practice (GMP) knowhow for setting up pharmaceutical plants to companies producing plasma, special enzymes and other biological substrates and even stem cell products for the repair of bones, cartilage or muscular tissue. Service providers such as good clinical practice (GCP) specialists help pharma or biotech companies and clinical trial units at hospitals to manage local or multicentre clinical trials to the highest regulatory standards. Close cooperation between universities, research centres and industry as well as highly trained and experienced local scientists, experts and technicians ensure the highest quality of products and services. Serving the Pharmaceutical Value Chain With an established and highly integrated industry, offering products and services underpinning production processes, Austria is in a strong position to compete in the world marketplace for globalised life sciences. To reinforce this already strong position, Austria recently founded two world-class research centres as an engine for enterprise: the Austrian Centre of Industrial Biotechnology (ACIB) and the Research Centre Pharmaceutical Engineering (RCPE). The ACIB, with locations in Vienna, Graz and Innsbruck, focuses on coordinating the applied biotechnological research of seven Austrian universities and around 30 international companies. The Research Centre Pharmaceutical Engineering GmbH (RCPE) is an interdisciplinary research institute in the area of pharmaceutical process and product development located 16 INTERNATIONAL PHARMACEUTICAL INDUSTRY

in Graz. The RCPE focuses on the development and production of pharmaceuticals using efficient, science-based methods, as well as their manufacturing processes. The success of the RCPE has already resulted in its first spinoff, Pharmaceutical and Regulatory Services, which is a full service provider for pharmaceutical regulatory affairs. These research centres reflect the pioneering spirit of companies across Austria, active in pharmaceutical production. The many innovative technologies make processes more efficient and less costly, and increase functionality, giving Austria the edge on the global stage. With established drug developers comes a need for outside support from suppliers and service providers. Austria has met the challenge with

a plethora of businesses such as suppliers of clean rooms, preclinical research facilities, reagents, prion testing, essential basic materials including glassware, right up to GMP plants and GCP quality assurance. There is enough diversity in the Austrian industry to build supplier networks that are never more than one flight-hour apart and can truly be labelled â&#x20AC;&#x153;Made in Austriaâ&#x20AC;?. The following are just some of the really dynamic enabling technology companies supporting the whole value chain of pharmaceutical production in Austria. Microinnova launched the Flow Miniplant as a tool for development and small-scale production in pharmaceutical and fine chemical companies - resulting in significant time and cost savings for clients. The engineers at Microinnova Summer 2012 Volume 4 Issue 3

Regulatory & Market Place have constructed manufacturing equipment in continuous flow mode for companies like Novartis, Roche, SigmaAldrich and Sandoz. A current collaboration including EvonikDegussa and the IMM Mainz are aiming to design the chemical plant of the future. For those companies looking to outsource production, Austria has its own home-grown contract manufacturing organisations (CMOs) as well. EUCODIS Bioscience, for example, is an application-driven, enzyme-engineering company with customers in the pharmaceutical, biotechnology, diagnostics and other industries. The company manufactures and markets over 50 well-characterised enzymes worldwide with a renowned customer portfolio, including Sandoz. piCHEM is a top producer of high-performance chemicals used in medical, biochemical, and pharmaceutical R&D. Montavit is another GMPcertified manufacturer focusing on sensitive plant extracts as well as developing and producing their own line of medicines. At the very end of the pharmaceutical value chain, when products are ready to be positioned for the market, are companies such as Stoelzle Oberglass. The companyâ&#x20AC;&#x2122;s goal is to work with customers to produce innovative packaging solutions for current market trends that meet customer demands, whether it be for lightweight glass bottles, or demanding shapes, as well as specific sealing solutions. In close cooperation with Stoelzle, Tubex is one of the leading companies in the packaging industry and specialises in the production of aluminiumaerosol cans, aluminium tubes and plastic tubes. The Austrian site is one of the most modern aluminium tube plants in the world. Over 140 highlymotivated members of staff bring in all their know-how to manufacture aluminium tubes in different sizes and makes for the pharmaceutical market. Advancing Life Science at the Heart of Europe The life science industry in Austria is fully diversified with companies 18 INTERNATIONAL PHARMACEUTICAL INDUSTRY

large and small as well as a number of multinational companies headquartered in or with facilities here - though it is small to mediumsized companies that predominate across a whole range of disciplines. The truly remarkable factor in the Austrian life science scene is the degree of cooperation between producers, networks of suppliers and service providers all within a short distance, meaning that the results are truly â&#x20AC;&#x153;Made in Austria!â&#x20AC;? Austria has over 210 life science companies employing approximately 11,500 people. Over 90% of these are small and medium-sized companies.

Additionally, there are around 600 companies acting as suppliers to the industry for components and services, meaning that in many cases the value chain is entirely Austrian. In addition to well-known large multinationals (including Sandoz, Roche, Greiner Bio-One, Med-EL, Baxter and Otto Bock, Boehringer-Ingelheim), many of the other companies are themselves market leaders and are well known to global industry experts. For a small country with a population of just over 8 million, that marks a very significant contribution to life science in Europe. It is also a key component of the Austrian economy and one that is Summer 2012 Volume 4 Issue 3

Regulatory & Market Place valued and supported by the national and regional governments. This thriving environment of successful and innovative companies has put Austria firmly on the European life science map. The combination of global players with research facilities in Austria and young, dynamic start-ups in close cooperation with excellent universities creates an ideal environment for the development, growth and prosperity of the Austrian life science industry. The average Austrian biotechnology company is just seven years old, yet the industry has reached a critical mass within a very short time period and has grown into a major economic force. Austria – the Place to do Business In recent years, the strength of the growing life science sector has been reflected in the increasing interest shown by international business in doing deals with companies in Austria. And it is not just the proliferation of corporate deals that shows Austria in a good light; the country is also proving to be an attractive location for operations and significant inward investment for a number of multinational companies. Since 1982, the American group Baxter has been running its biggest subsidiary outside the US and its most important research site worldwide in Austria, with over 4000 employees. More than 900 scientists from home and abroad are currently working on the discovery of new drugs as well as the enhancement of medications that are already on the market. Baxter’s biosciences division in Austria focuses on developing and manufacturing biotechnological and biopharmaceutical therapeutics, of which 90 per cent are being exported. The German prosthetics manufacturer Otto Bock has also located a key facility in Austria, improving and restoring the lives of patients all over the world. These investments come on top of a whole series of international venture capital investments in Austrian life science companies. In 2010, funding of the Austrian life science sector totalled around €79 million from venture capitalists, private investors, grants, loans and other contributions. 20 INTERNATIONAL PHARMACEUTICAL INDUSTRY

A further €37 million was invested in Austrian medical technology companies. So why does Austria have such a great reputation for life science investment? The New York-based Reputation Institute has conducted a study that shows Austria is ranked ninth out of 50 countries in terms of trust, reputation, admiration and positive image. Also, the Mercer Study 2011 ranked Vienna as the city with the highest standard of living in the world. Austria’s extensive infrastructure and its enviable location at the geographic heart of Europe are also significant reasons. Add in additional factors such as the excellence of the Austrian workforce, the renowned quality of its education system and the top-tier status of its research institutions, and it is easy to see how many companies find Austria a compelling location for doing business. Infrastructure Great infrastructure is a key driver of the life science industry in Austria. Across Austria, a succession of science parks, incubators and tech transfer initiatives has ensured the growth of the indigenous industry as well as attracting a plethora of established multinational concerns. This is an ongoing process with a constant stream of new initiatives and investments being added to the existing ones. Austria’s World-Class Research Base Austrian medical universities focus on state-of-the-art R&D for new therapeutic substances with more than 55,000 people involved in medical research. Besides the medical universities, there are also renowned academic bodies that significantly contribute to life sciences research in Austria and that have generated many successful life science spinoffs. BOKU, for instance, is the main university in the country for natural resources and life sciences, while the Graz and Vienna Universities of Technology play an important role in life science research and in the training of skilled engineers. Alongside the universities, the

Austrian Academy of Sciences (AAS) is the leading organisation promoting non-university-based academic research institutions in Austria. The AAS’s life science research portfolio is conducted in institutions including the Research Centre for Molecular Medicine (CeMM), the Gregor Mendel Institute of Molecular Plant Biology (GMI), the Institute for Biomedical Aging Research (IBA) and the Institute of Molecular Biotechnology (IMBA). It is also worth noting the Institute of Science and Technology Austria (IST Austria), a PhD-granting institution located in the Vienna Woods and one of the principle locations of research in the natural sciences including bioscience and the Austrian Institute of Technology (AIT), Austria’s largest non-university research institute with a research focus on grand societal challenges. Research at the Institute of Molecular Pathology (IMP) spans a wide range of topics. The common goal in all of these areas is to elucidate the mechanisms and principles that underlie complex biological processes. From Bench to Boardroom Arguably, much of the recent investment success would not have been possible without a government that backs innovation and supports outstanding academic research. The resulting cutting-edge research at Austrian universities is the source of technology transfer to the many start-up companies proliferating in the Austrian life science scene. The Austrian government is committed to this burgeoning sector and is helping to foster a business environment that allows these young spin-off companies to thrive. With R&D investment in 2011 at an all-time high of 2.79% of GDP, Austria already invests far more into R&D than the EU average. The government’s new strategy for research, development and innovation aims even higher: to make Austria an innovation leader within the European Union and to raise the share of R&D investment to 3.76% of GDP by 2020. There is also a very supportive and attractive tax regime, an R&D cash premium of 10% and a maximum corporate income tax of 25%. Summer 2012 Volume 4 Issue 3

Regulatory & Market Place Sources of Finance The most innovative ideas in life sciences only make a difference once they’ve survived the journey from the laboratory bench to commercial launch. However, that journey requires careful nurturing in a supportive funding environment. Austria has developed a wide range of national and regional funding schemes to back new companies embarking on this journey towards business ´ success. Focusing on start-up ideas in biotechnology and medical devices, austria wirtschaftsservice, the Austrian government promotional bank, provides financial support through two custom-designed funding programmes: “PreSeed LISA” and “LISA Seedfinancing”. PreSeed LISA provides funding for the critical phase before a life science company is actually set up. Costs relating to the implementation of the science and the development of a business plan for a project can be funded with non-refundable awards of up to €200,000. Setting up an innovative, internationally competitive high-tech company needs significant knowhow, courage and capital. LISA Seedfinancing supports this start-up phase by providing up to €1 million, combined with business advice. This seed financing is then refunded to austria wirtschaftsservice at the point a company is making a profit or is sold. Importantly, in contrast to bank or other more traditional loans, there is no requirement for customary securities. However to qualify for this funding a company must be partly and suitably funded through private capital. Other austria wirtschaftsservice initiatives to support life sciences include guarantees and loans. In addition there is also a wide range of regional funding available. A Country of Many Parts Region by region, the life science industry is spread across Austria from the capital Vienna to the powerhouses of Styria, Tyrol and Upper and Lower Austria. Each region has its own special culture and particular strengths that are reflected in the industries located there. Organised 22 INTERNATIONAL PHARMACEUTICAL INDUSTRY

through regional life science clusters, LISA represents companies in the therapeutic, medical technology and diagnostic sectors as well as providers of enabling technologies and related service companies located in the following regions: Committed to the Prosperity of the Austrian Life Sciences The national body Life Science Austria (LISA) promotes the Austrian life science sector on the international stage and is the first point of call for enquiries relating to it. Organised through the regional life science clusters, the organisation represents companies in the therapeutic, medical technology and diagnostic sectors as well as providers of enabling technologies and related service companies located in the following regions: • Lower Austria (ecoplus), • Styria (, • Tyrol (Cluster Life Sciences Tyrol), • U pper Austria (Health Technology Cluster), • Vienna (LISAvienna) LISA is run by austria wirtschaftsservice on behalf of the Federal Ministry of Economy, Family and Youth. The organisation is committed to the development, growth and prosperity of the Austrian life science industries as a leading component of the Austrian economy. On the international front, they work towards Austria becoming known worldwide for the excellence of its life science sector. In the home market, LISA is a resource for all life science companies within, and all companies wishing to relocate to, Austria. They support companies in making connections locally and further afield. They also play a big part in helping new companies get started through their Preseed and Seed financing funding programmes. Every two years, LISA runs a life science business plan competition BOB Best of Biotech as our way of fostering a culture of excellence. For more information, please visit:

Sonja Polan is International Marketing Manager of LISA - Life Science Austria. Previously she worked as Account Manager at the European Commission, mainly in charge of marketing telecommunications policies to the European public. The most important campaigns were for the newly established roaming regulation and telecom reform. Before the European Commission, she worked for a major Brussels-based European public affairs consultancy, and also had short stints with the European Parliament and with trade federations. She speaks German, English and French. Email:

Summer 2012 Volume 4 Issue 3

Regulatory & Market Place Pittsburgh, PA.


Regulatory & Market Place

Extending your Patent on a New Application for an Old Drug Summary The period that is necessary to obtain a marketing authorisation (MA) to bring a new drug onto the market in the European Union significantly reduces the effective patent protection of that drug and, hence, the possibility to recover the high research and development costs. To compensate for that loss, the European Union member states provide supplementary protection certificates (SPCs), which may extend the patent life of certain human and veterinary pharmaceuticals for up to five years. A further six-month extension is possible for medicinal products for paediatric use. Since the introduction of SPCs in 1993, this type of patent life extension has become key in the pharmaceutical industry. Until recently, it was generally accepted that an SPC could only be awarded for a patent on a new active ingredient, and that patents on new applications for “old” active ingredients (i.e. further medical use patents) were excluded from an extension. However, on 19 July 2012, the Court of Justice of the European Union provided a ruling that suddenly allowed SPC extensions for further medical use patents. In this article we will outline what SPCs are, how this recent ruling in the so-called Neurim case fits with earlier case law, and which types of further medical use patents may receive an extension. SPC Regulation Regulation (EC) No 469/2009 of 6 May 2009 governs the granting of supplementary protection certificates for medicinal products, wherein Article 3 details the requirements for obtaining an SPC: A certificate shall be granted if, in the Member State in which the application (…) is submitted and at the date of that application: a) the product is protected by a basic patent in force; b) a  valid authorisation to place 24 INTERNATIONAL PHARMACEUTICAL INDUSTRY

the product on the market as a medicinal product has been granted in accordance with Directive 2001/83/ EC or Directive 2001/82/EC, as appropriate; c) the product has not already been the subject of a certificate; d) the authorisation referred to in point (b) is the first authorisation to place the product on the market as a medicinal product. In short, without going into too much detail, ‘medicinal product’ refers to any substance or combination of substances which may be administered (thus including excipients etc.), ‘product’ refers to the active ingredient(s) and ‘basic patent’ means a patent that claims a product, a process to obtain a product or the application of a product, and that forms the basis of the SPC application. To calculate the duration of an SPC, the period between the date of filing the basic application and the date of the first authorisation to place the product on the market in the Community is reduced by five years, i.e. (MA - patent filing) - 5y. The maximum duration of an SPC is five years, but an additional six-month paediatric extension may be obtained, which brings the maximum SPC term to 5.5 years for such pharmaceuticals. Earlier Case Law Before the Neurim ruling, the SPC regulation was considered to exclude extensions for further medical use patents, not least due to a number of rulings from the Court of Justice of the European Union (CJEU). Herein below, we briefly summarise the CJEU judgements that concern SPC applications on new applications of a known active ingredient, all of which were ruled to the disadvantage of the SPC applicant. In case C-31/03, Pharmacia Italia wanted to obtain a German SPC on

its patent for the active ingredient cabergoline, for which an authorisation as a medicinal product for human use was issued in the Netherlands in 1992. However, an earlier Italian authorisation had been issued in 1987 for the veterinary product Galastop, which contains cabergoline. The German patent office rejected the application on the basis that the first MA was the veterinary MA that was obtained before 1988. According to Article 19 of the original SPC regulation (which is a transitional provision that is not present any more in the current 469/2009 regulation), the first MA should be obtained after 1988 in Germany. Pharmacia appealed the decision, arguing that the first MA must be the first MA for a medicinal product for human use, and the German court referred the issue to the CJEU. In its argumentation the CJEU states that “the decisive factor for the grant of the certificate is not the intended use of the medicinal product and (…) the purpose of the protection conferred by the certificate relates to any use of the product as a medicinal product without any distinction between use of the product as a medicinal product for human use and as a veterinary medicinal product”. Therefore, the veterinary MA dating from before 1988 precluded Pharmacia from getting a German SPC for the human use. In case C-202/05, Yissum Research and Development Company of the Hebrew University of Jerusalem applied for an SPC extension on its patent for a composition comprising the active ingredient calcitriol for use in the treatment of skin disorders. The UK patent office refused the application because, contrary to Article 3(d), earlier MAs had been given for medicinal products containing the active ingredient calcitriol. The cited MAs concerned other therapeutic applications, namely management of hypocalcaemia and treatment of chronic renal failure Summer 2012 Volume 4 Issue 3

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Regulatory & Market Place or post-menopausal osteoporosis. Yissum brought an action against that decision, arguing that “product” in their case was “calcitriol for the topical treatment of psoriasis”, which led to a referral to the CJEU for a preliminary ruling on, amongst other things, “does the application of the therapeutic agent play any part in the definition of “product” for the purpose of the Regulation?”. In its order, the CJEU stated that “in a case where a basic patent protects a second medical use of an active ingredient, that use does not form an integral part of the definition of product”. C-195/09 (Synthon) and C-427/09 (Generics) are two cases that are very similar to each other. In Synthon, the active ingredient memantine received a marketing authorisation for the treatment of Parkinson’s disease, which, under an old German legislation, didn’t require testing for safety or efficacy. Merz obtained a European patent for memantine hydrochloride based on the further medical use of memantine to treat Alzheimer’s disease. Merz later obtained an SPC on the patent, referring to the MA for Alzheimer’s disease as the first MA. In Generics, the active ingredient galantamine was on the market for the treatment of poliomyelitis; also under old legislations that didn’t require safety and efficacy testing. Synaptech obtained a European patent for the use of galantamine in the treatment of Alzheimer’s disease. Subsequently, it was granted an SPC on the patent, listing the MA for galantamine in the treatment of Alzheimer’s disease as the first MA to place the product on the market. In both instances, a generic producer (Synthon BV and Generics UK Ltd.) brought action before the UK court, which referred questions to the CJEU. In its response, the CJEU provided the same ruling in the two cases: “a product, such as at issue in the main proceedings, which was placed on the market in the European Community as a medicinal product for human use before obtaining a marketing authorisation in accordance with Council Directive 65/65/EEC (…), in particular, without undergoing safety and efficacy testing, is not within the 26 INTERNATIONAL PHARMACEUTICAL INDUSTRY

scope of Regulation No 1768/92, as amended, and may not, therefore, be the subject of a supplementary protection certificate”. The Neurim Case Melatonin is a natural hormone and its use in sheep to regulate seasonal breeding received an MA in 2001. Neurim Pharmaceuticals discovered that appropriate formulations of melatonin could be used as a medicine for insomnia. It received a European patent for its formulation and an MA to market the medicinal product for

human use. Based thereon, Neurim applied for an SPC. The application was refused by the UK patent office because of the earlier veterinary MA for seasonal breeding, i.e. contrary to the requirement of Article 3(d) that the MA is the first MA to place the product (in this case melatonin) on the market. Neurim defended its case up to the Court of Appeal by arguing that the relevant MA is the one that concerns the product for which the SPC application is made, and that no SPC application was filed in relation to the veterinary MA. Summer 2012 Volume 4 Issue 3

Drug Discovery/ Development & Delivery


Regulatory & Market Place The Court of Appeal sympathised with Neurim’s situation since “commercially, medically and legally there is a vast expanse of clear blue water between the parties’ products and legal rights. None of the work done by Hoechst helped Neurim at all – it may indeed have hindered them because the regulator would naturally have been concerned about possible side effects.” and “if Neurim are wrong, then the Regulation will not have achieved its key objects for large areas of pharmaceutical research: it will not be fit for purpose. Whether that is so or not is clearly a matter for the EU’s highest court”. Largely unexpectedly, the CJEU agreed with Neurim and provided a new teleological interpretation of the SPC regulation. In its grounds, the CJEU noted that “if a patent protects a therapeutic application of a known active ingredient which has already been marketed as a medicinal product, for veterinary or human use, for other therapeutic indications (…), the placement on the market of a new medicinal product commercially exploiting the new therapeutic application of the same ingredient, as protected by the new patent, may enable its proprietor to obtain an SPC”. In other words, a human or veterinary MA for a first medical use application does not preclude one from getting an SPC for a second medical use application. To determine the duration of the SPC, the first MA “refers to the MA of a product which comes within the limits of the protection conferred by the basic patent relied upon for the purposes of the application of the SPC”. Thus, in general, the duration will be determined by the date of filing the further medical use patent and the “further medical use MA”, as the “first medical use MA” is normally not covered by a further medical use patent. Conclusions Since the grounds of Neurim as well as Pharmacia make clear that there is no difference if an MA is for a veterinary or human use, the Neurim ruling certainly opens the door for further medical use patent extensions, regardless of the species to which the 28 INTERNATIONAL PHARMACEUTICAL INDUSTRY

first or further medical use application relates. Nonetheless, a number of questions remain. In essence, Yissum also applied for an SPC application for a further medical use patent, but they did not get a favourable ruling. Complementarity of the two rulings can only be explained by interpreting the Yissum ruling in its narrowest sense, i.e. only as an interpretation of the definition of product. Thus, one might wonder if Yissum would have got their SPC if they had argued that the “first MA” related to the second medical use, instead of the “product”. The Synthon and Generics rulings may also be explained in light of Neurim by such a narrow interpretation, namely that further medical use patents are only excluded from an SPC if the active ingredient was already on the market before receiving an MA according to the specified Directive. Nonetheless, that interpretation creates a seemingly unfair criterion for determining if a patent is eligible for an SPC or not. After all, how can one make sense of the fact that putting the active ingredient for the first medical use application on the market before or after obtaining an MA according to the specified Directive would make such a big difference for an applicant wanting to exploit a further medical use application? The fact that the applicant for the further medical use SPC already benefited from the first medical use marketing in both Synthon and Generics does not provide for an answer. Indeed, the Neurim ruling says “the answers (…) would not be different if (…) the product covered by the first MA of the corresponding medicinal product is within the scope of protection of a different patent which belongs to a different registered proprietor from the SPC applicant”. All in all, it remains to be seen if and how the Synthon and Generics rulings will be applied after the more favourable Neurim ruling. One further question that is still open for discussion is what if an earlier medical use patent already obtained an SPC? This was not the case in Neurim, and, accordingly, no issues were raised in that instance. But Article 3(c) clearly states that one of the requirements is that “the

product has not already been the subject of a certificate”. Since the product definition in that article does not include the medical application (as ruled in Yissum), this appears to exclude the possibility to be granted an SPC for a further medical use application if an earlier medical use patent for the same active ingredient already obtained an SPC. Again, it appears to be unfair to a further medical use innovator to make his SPC application dependent on what the first medical use application did or did not get. To conclude, the door for extending the patent life of further medical use patents has been opened by the Neurim ruling, although it will likely take further case law from the CJEU to determine the full scope of the SPC legislation. Furthermore, further amendments from European Union legislators may be needed to arrive at a legislation that allows for the grant of patent extensions for pharmaceutical innovators on a fair and clear basis. Nonetheless, given the importance of SPCs in today’s pharmaceutical industry, we expect that the current Neurim decision will lead to a much larger number of SPC applications for further medical use patents.

Stijn Lagaert graduated in 2006 as a master in Bioscience Engineering at the University of Leuven (KU Leuven). He subsequently followed a PhD program at the Division of Gene Technology (KU Leuven), where he acquired a detailed knowledge on biotechnology and its interface with food science. In 2011, he joined LC Patents, where he is involved in drafting and prosecuting patents, as well as analyzing freedom-to-operate and patentability. His focus areas in intellectual property are Biotechnology and Medicinal Chemistry. In addition thereto, he is training for the European Qualifying Examinations in order to become a qualified European Patent Attorney. Email:

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Regulatory & Market Place

Health – Made in Germany The market for healthcare is gaining steadily in global importance and offers great promise for innovation, growth and employment. Demographic developments, increasing health awareness and even new products and technologies themselves, are driving the dynamics of further growth. The worldwide market for healthcare holds great potential for further international co-operation, including with Germany’s healthcare industry – a top international player. Through the initiative “Health – Made in Germany”, the Federal Ministry of Economics and Technology (BMWi) is supporting German healthcare companies and their global partners as they exploit opportunities to co-operate and do business. Why Germany? Germany was among the top five countries in the world in terms of market size in 2010, with a generated turnover of USD 35 billion1. Meanwhile, government statistics indicate that around 900 companies – most of them small or medium-sized firms or SMEs – are active in pharmaceuticalsrelated fields2. While most of them are trading on foreign markets, they also strengthen the country’s domestic economy. Germany’s fermentation capacity is second only to the United States. That has made the country Europe’s biggest producer of biopharmaceuticals, with sales of EUR 5,2 billion in 20103. Germany’s current status at the top of the ranks in the pharmaceuticals and biotechnology sectors of healthcare is no coincidence. A long tradition in producing drugs, of academic excellence, an innovative business climate, public support for research and development and geographic advantages have helped secure its leading position. Striving for Innovation Germany’s industrial and institutional 30 INTERNATIONAL PHARMACEUTICAL INDUSTRY

innovative drive is reflected in an impressive array of registered patents. In 2010, Germany led Europe with close to 12,000 patents granted in the field of pharmaceuticals and medical biotechnology (10,384 pharmaceutical, 1228 biotechnological) 4. Bearing in mind the fact that the German pharmaceutical industry spent in 2009 about 13.7% percent of their sales on

research and development, the high number of patents does not come as a surprise5. What is more, the German government has identified life sciences as a decisive factor in economies of the 21 st century, and has made available a variety of public subsidies for R&D in this field. Between 2012 and 2015 the German government will Summer 2012 Volume 4 Issue 3

Regulatory & Market Place invest EUR 8.3 billion in what is known as the “High-Tech Strategy”6. Another part of the “High-Tech Strategy” is a competition aimed at identifying “leading-edge clusters”. The 15 winners of the three-round competition are each provided with EUR 40 million in funding by Germany’s Federal Ministry of Education and Research (BMBF) for a period of five years to implement their strategies. In addition to R&D grants, the clusters will also be eligible for reduced-interest loans and special partnership programmes 7. Beyond that, the federal states have their own R&D grant programmes. Most are designed for SMEs and focus on specific industry clusters. In line with improving R&D infrastructure, the government is promoting clinical trials because these are seen as drivers of innovation and a major part of quality assurance in medicine. The government is also trying to improve conditions for clinical trials in Germany through funding and by setting up various interdisciplinary centres for clinical research. The aim is to speed the transfer of research findings into practice. The strategy has proven successful, with Germany recently leading Europe in the number of sites in clinical trials in Europe 8. One key to Germany’s success in this area is cost. While data quality is on a par with the US, costs are up to 50 per cent lower in Germany9. That gives Germany a competitive advantage in terms of lower costs and enhanced levels of expertise and quality. The recently published “Guide to Contract Research in Germany” can assist you in locating the right partners conducting clinical trials in Germany ( Institutional and Academic Research Thriving Germany has held a leading position in drug development for more than a century. Some of the world’s most sought-after medications were invented in the country. Also home to some of the most successful traditional pharmaceuticals companies, universities and research organisations, Germany’s outstanding success in the field of 32 INTERNATIONAL PHARMACEUTICAL INDUSTRY

healthcare can in part be attributed to well-established and traditional cooperative ties between companies, research organisations and the academic community. The pharmaceuticals industry in Germany also benefits from internationally renowned scientists who come to the country to carry out world-class research at institutes and universities or the companies themselves. The number of people employed in decided biotechnology research companies is a lot higher than in any other industry – with around 45 per cent of the German workforce holding a university degree10. The University of Freiburg and the University of Frankfurt-am-Main are globally known for the work they do in the pharmaceuticals field. Like their other German counterparts, they cooperate closely with a variety of

research institutes in order to advance basic and applied research as well as get new products onto the market. Germany’s biggest research organisation, the Helmholtz Association, has an annual research budget of EUR 2.7 billion, and employs 26,500 staff. Another important organisation is the Max Planck Society, which has more than 80 institutes dedicated to the full range of basic research in, among other subjects, biology, technology and medicine. The Leibnitz Association is an inter-disciplinary scientific community, which has 86 research institutes across the country, of which 25 specialise in life sciences. Last but not least, Germany’s Fraunhofer Gesellschaft is Europe’s largest applied research institution. It employs a staff of 18,000 in 80 research facilities 11. Summer 2012 Volume 4 Issue 3

Regulatory & Market Place Europe’s Fermentation Centre Germany became involved in biotechnology early on and has evolved into one of the world’s leading biotech hubs. With a fermentation capacity of 675,000 litres, Germany is Europe’s largest producer of biopharmaceuticals, and second only to the US in the world 12. A number of clusters relevant to the biotechnology industry have fuelled this development. Among the largest are located near Munich, Berlin and in the Rhine-Neckar triangle and the Ruhr area. A major biotechnological hotspot is the BioRegion Ulm, which is home to one of Europe’s largest mammalian cell culture facilities in Europe13. Most genetically manufactured drugs are produced here. This cluster is made up of 60 pharmaceutical, biotechnological and medical technology companies. In addition to the healthcare industry’s quantitative strengths, high standards for quality and reliability set German pharmaceutical and biotechnological products apart from their competitors. Maintenance of the highest technical standards as well as the corresponding regulatory requirements in production and quality control are a matter of course. Optimal product quality is additionally guaranteed by the knowhow and skill of a highly-qualified labour force. Dynamic Growth on a Global Scale Demographic developments, increased health awareness and fast-paced growth in countries like China, Brazil, Russia and India is spurring the call for products and services. What is more, an increasing population of elderly people with chronic problems such as diabetes or cardiovascular disease is driving the call for healthcare goods and services in the industrialised world in particular. Given those factors, demand is diversified as well, leaving consumers searching for providers who can deliver the latest technologies and products to meet a wide range of healthcare needs14. Whether it is pharmaceuticals, medical technology, medical biotechnology or health-related services, German industry responds to this challenge with innovation 34 INTERNATIONAL PHARMACEUTICAL INDUSTRY

and product development. New methods for diagnosis and treatment are facilitating the therapy of as yet incurable diseases, which in turn enhances the life expectancy, and thus extends the utilisation of the healthcare system15. As a global export leader, Germany’s pharmaceutical sector has already reaped advantages from these market developments. A study by a major international consulting firm Prognos AG indicates the sector’s export volume quadrupled between the years 2000 and 2008 to EUR 45.2 billion. The consultants say that they expect the pharmaceuticals market’s exports to grow further at an aboveaverage rate of 6.3 per cent until 2015 16. Currently, the European market is still the main destination for German exports, followed by the US, which accounts for more than half of the exports going to destinations outside the EU 17. Analysts who are experts in innovation say that densely populated countries like India, China and Russia, as well as those in Latin America, such as Brazil and Argentina, will have greater influence on Germany’s exports in future. Fostering Global Cooperation BMWi has established the initiative “Health – Made in Germany” in order to support the healthcare industry in its efforts to expand activity in markets around the globe. The main aim of the programme is to foster bilateral trade in the healthcare market and support German industry in entering emerging markets. Strong partners like federal ministries, professional associations and Germany’s federal states co-operate through it to achieve a common goal. Health – Made in Germany is implemented by Germany Trade & Invest, the German government agency for foreign trade and inward investment. The initiative has been organised into four main healthcare industry sector working groups: pharmaceuticals, biomedical, medical technology and telemedicine and health-related services. Among the members of the different groups are 14 different branch associations. This structure has been chosen to establish

a channel of direct communication with partners within each sector in order to allow industry a constant dialogue to co-ordinate and direct the goals and activities of the initiative. The inception of Health – Made in Germany centrally bundled all activities related to foreign trade in the healthcare industry. Its internet portal is the first point of contact for foreign partners and German companies who are interested in international business. The site offers information on current initiative activities including projects, tenders, grants and market data. Furthermore, the strengths of the German healthcare economy are presented in a way that makes it easy for foreign partners and decision-makers to access and enjoy the advantages of cooperating with German companies. The initiative has already drawn global attention. Until now, it has been active on four continents and in more than 16 countries around the globe. At the same time, the pharmaceuticals and biotechnology working groups have started focus projects in Russian and India respectively. For the pharmaceuticals and biotechnology market, the respective working groups have started two focus projects in Russia and India. At the moment, these countries are currently engaging in political efforts to create a more suitable business climate for German companies. The Russian pharmaceuticals market holds vast potential and enormous opportunity for German drug-makers. In the past years though, they have surprised the Russian administration with mostly short-term regulatory acts, which could not be met by the German companies in the given time-frame. Health – Made in Germany has attempted to resolve this problem by setting up what is known as a pharmaceuticals “earlywarning system” in co-operation with a Russian market research company. It includes a regularly published newsletter in Russian and German which details new regulations on the Russian pharmaceuticals’ retail market, including items on licensing and reimbursement that are of interest to German firms doing business with Summer 2012 Volume 4 Issue 3

Regulatory & Market Place Russia. The publication also clarifies other implications Russian regulatory decisions could have for foreign drug firms interested in commerce there. Another major issue is the strengthening of cooperation between India and Germany in the field of life sciences. In particular, the pharmaceutical and the biotechnology sector are in the focus of attention. The “Indo-German Healthcare Dialogue”; a platform that was initiated last year by the initiative Health – Made in Germany will help to achieve this target. With the aid of the platform and in cooperation with the diplomatic mission, Indian and German entrepreneurs get the opportunity to gather on a regular basis. During their meeting, questions of research cooperation and innovation represent key issues for companies joining the Indo-German Healthcare Dialogue. Together they try to lobby for an adjuvant, supporting environment which is indispensable to foster partnerships and bilateral collaboration in this sector. Locating Partners Health – Made in Germany has, in addition to its website, published a series of directories that give an overview of part of Germany’s pharmaceutical and biotechnology sectors and their capacities. Created to simplify partner access to the wealth of expertise and experience Germany has to offer, the Guide to Contract Research in Germany, and the German Biomanufacturing Guide can be retrieved, like many other reports and documents, free-of-charge from the initiative’s website. The Guide to Contract Research includes a listing of more than 120 companies, mainly SMEs. It also provides company profiles and graphic tables that describe the types of research done by each company. The German Biomanufacturing Guide serves as an orientation aid for the complex, rich and very diverse German biomanufacturing sector. Nearly half of all German biomanufacturing activities are currently orientated towards medicine and healthcare. Most medical biotech companies employ less 36 INTERNATIONAL PHARMACEUTICAL INDUSTRY

than 50 people, yet frequently cooperate and interact with much larger stakeholders in industry, academia and with researchers at globallyknown private and public institutes and organisations. Compiled in early 2011 from a survey of the country’s biomanufacturing companies, the guide contains profiles of more than 30 firms that vary in terms of their focus, staff size and fermentation capacity. While the German pharmaceuticals and biotechnology sectors have already realised a considerable amount of their potential in the domestic and foreign markets, there is still plenty of room for new growth. If you are interested in further information on the German healthcare market, ongoing projects, or current healthcare news, a visit to com is highly recommended. References 1. Verband forschender Arzneimittelhersteller e.V. (vfa). Statistics 2011. Die Arzneimittelindustrie in Deutschland, p. 45 (2011). 2. B  undesverband der Pharmazeutischen Industrie e.V. (BPI). Pharma-Daten 2011, p. 8 (2011). 3. V  erband forschender Arzneimittelhersteller e.V. (vfa). Statistics 2011. Die Arzneimittelindustrie in Deutschland, p. 52 (2011). 4. B  undesverband der Pharmazeutischen Industrie e.V. (BPI). Pharma-Daten 2011, p. 1617 (2011). 5. B  usiness Monitor International. Germany Pharmaceuticals and Healthcare Report Q2 2012, p. 30 (2012). 6. w, visited on 09 August 2012. 7. F  ederal Ministry of Education and Research (BMBF). Germany’s Leading Edge-Cluster (2012). 8. F  ederal Ministry of Education and Research (BMBF). Rahmenprogramm Gesundheitsforschung der Bundesregierung, p. 22 (2010). 9. D  eutscher Bundestag. Innovationsreport Biomedizinische Innovationen und klinische









Forschung Wettbewerbs- und Regulierungsfragen, p. 23 (2009). w w w. b i o t e c h n o l o g i e . d e / B I O / Navigation/DE/%20Hintergrund/ studien-statistiken,did=61968. html, visited on 06 August 2012. w w w. r e s e a r c h - i n - g e r m a n y. d e / main/research-landscape/41834/ rpo.html, visited on 10 August 2012. H  ealth - Made in Germany. German Biomanufacturing Guide, p. 5 (2011). w w w . b i o - p r o . d e / m a g a z i n / wirtschaft/archiv_2007/index. html?lang=en&artikelid=/ artikel/01615/index.html, visited on 07 August 2012. K  artee, J., Neumann, K. Weltweite Gesundheitswirtschaft – Chancen für Deutschland, p. 4-11 (2011). M  eidenbauer, T. Das Wachstum der Gesundheitsausgaben – Determinanten und theoretische Ansätze. Bayreuth: Institut für Volkswirtschaftslehre, p. 11 (2005). B  oehmer, M., Limbers, J., Zweers, U. Wie stark ist der Exportmotor Pharma? Kurzstudie zum Pharmastandort Deutschland, p. 9 (2009). B  undesverband der Pharmazeutischen Industrie e.V. (BPI). Pharma-Daten 2011, p. 13 (2011).

Marion Luekemann is working as a Senior Manager for the initiative “Health - Made in Germany”. She supports foreign enterprises seeking partnerships with German healthcare institutions and companies as well as German healthcare firms that are looking for international expansion. Prior to joining Germany Trade & Invest, Marion worked in the pharmaceutical industry for six years. She holds a degree in international cultural and business studies as well as an MBA in international management. Marion gathered several years of work experience in Russia and Poland. Email:

Summer 2012 Volume 4 Issue 3

Drug Discovery/Development & Delivery

Novel Factories for Novel Vaccines

Summary Vaccination is considered one of the most successful strategies for fighting infectious diseases, an area where there still remain significant unmet medical needs. In the past, it has been difficult for business to find opportunities in this area due to the long development times and to the risks involved in testing, for what could be relatively low returns, as many of the development opportunities were in low income countries. This makes it hard to build the right business proposition – right product price – that could bring a return on the high-risk investment. A relief in this status has been observed recently: the establishment of clear needs in terms of vaccines at global level has given the right degree of predictability to commercial entities. This predictability removes some of the risk. Better business models that can be applied to these indications, better understanding of the disease mechanisms, along with application of novel technologies have enabled new vaccine projects that address diseases with great need. Globalisation has also meant that diseases are not “local”, and their ability to spread fast globally has been seen in recent years. The traditional methods for preparing vaccines are not suitable for the fast response which is needed to face global epidemics. This opens the opportunity to develop more efficient and better adapted platforms. Manufacturing platforms and technology have been aspects that have proven very relevant both in translating the science into a product, and in responding to the challenging need to control manufacturing costs. Faster, more flexible, less capitalintensive platforms are needed, that can fulfill the potential of rationally designed vaccines. The importance of understanding and including the right adjuvant has been increasingly recognised, triggering considerable 38 INTERNATIONAL PHARMACEUTICAL INDUSTRY

research in this area. The focus of this short review will be on recent alternative manufacturing platforms for novel vaccine production. Examples from the development pipeline of vaccines to Dengue fever, West Nile and Japanese Encephalitis virus will be used to illustrate the issues. Introduction Although vaccination is demonstrably one of the most efficient ways of fighting infectious diseases, there are still significant neglected needs for a number of diseases, despite recent important advances. For example, following the introduction of pneumococcal conjugate vaccine, an estimated 211,000 serious pneumococcal infections and 13,000 deaths were prevented during 20002008. Routine rotavirus vaccination, implemented in 2006, now prevents an estimated 40,000-60,000 rotavirus hospitalisations each year1. Launched in 1974, the Expanded Programme on Immunization (EPI) was first designed to deliver vaccines against diphtheria, tetanus, pertussis, polio, measles and tuberculosis (BCG). Together, these vaccines prevent close to 2.5 million deaths every year2. Diseases most prevalent in poorer parts of the world have lagged behind as targets for vaccine development, certainly because they are challenging to tackle, but also for lack of the right business model that provides an acceptable level of risk for the investment commitments that are required. On the other hand, recent years have provided interesting returns to vaccine manufacturers, with sales of vaccines reaching US$25.3 bn in 2010. This figure is predicted to increase at a compound annual rate of 9.3% during 2010–2015, to reach US$39.5 bn in 2015 as new product introductions continue and usage of current products expands further3.

Promising new areas for development include RSV and Dengue, but also S. aureus, and other nosocomial infections, and immunotherapies for neurological and metabolic diseases, as well as cancer. The changing environment, pressure from payers to cut costs, and the implications of certain decisions - for example, as already indicated by the insurance companies, the possible exclusion of nosocomial diseases from insurance repayment - have a real impact on business opportunities, and illustrate clearly how societal factors condition the opportunities beyond the science and investment. In order to represent a real business opportunity, the development of vaccines needs to have the right science, but politics and financing also need to come together. Politics have a role in the effectiveness of implementation and extent of vaccine campaigns, for example, in allowing access to vaccination centres and for follow-up. Making a Vaccine There are three main “science-based” components in a vaccine — antigens, adjuvants and delivery system/ administration schedule. Production platform and supply chain efficiency are two strategically enabling and differentiating factors. A recent analysis by Frost & Sullivan Figure 1: Matching manufacturing innovation to novel vaccine design technologies

Summer 2012 Volume 4 Issue 3

on the vaccine market4 highlights that the most significant technical challenges in the field of vaccine development at present are: selection of appropriate expression systems for recombinant or subunit vaccines, cold chain storage requirements, and production challenges with egg-based vaccines. This latter is triggering interest in developing new types of platforms, for example, the use of plants to produce virus-like particles (VLPs). Choice of manufacturing platform is strategically important in vaccine design, and will condition cost and ability to respond to need. Figure 1 depicts the interconnection between modernisation in both technology for antigen development and manufacturing units of operation. At one end are the traditional life attenuated vaccines, produced in the most traditional platforms, such as chickens’ eggs; at the other end of the spectrum are innovations in facility design and in unit operations that have enabled and are adapted to innovative recombinant technology platforms. Traditional steel-based technology represents high investment and low flexibility, whilst the more modern platforms, based on recombinant technologies and facilities that include single-use technologies, allow for greater manufacturing flexibility. Development of new antigens has also benefited from increased platform versatility, which have made possible rational vaccine design. An examination of the current vaccine R&D portfolio activity reveals that it is still highest in antigenic component development followed by delivery systems. Adjuvant research is highly fragmented, but is now being recognised by vaccine developers as essential for making vaccines with high efficacy. Adjuvants are now considered an area of opportunity for development, in particular for the newer indications in non-infectious contexts such as cancer, neurology and metabolic diseases, because of their interaction with the immune response. Adjuvant development is challenging, not only because the exact mechanisms by which stimulation of the immune system occurs, and the exact type of response sought, are still

Table 1: Adjuvants in use and in development

poorly understood, but also (and consequently) because the path to approval is long and challenging. Thus, often the better known adjuvants continue to be used, although some of the newer technologies such as use of oligonucleotides are finding good grounds to advance. The potential of adjuvants in permitting dose reduction or overcoming nonresponse clearly marks the importance of continuing to understand the role of these important vaccine components. Table 1 summarises the current adjuvants in use and in development 5. Amongst the most recent advances, two are highlighted for their originality: use of Lactococcus “shells”, or BLPs (bacterium-like proteins) being developed by the Dutch company Mucosis and the very early stage and intriguing use of the protozoan Tetrahymena thermophila and its ability to produce naturally occurring protein nanoparticles by Tetragenetics which the company markets as their G-SOME™ vaccine technology6. The Mucosis BLPs, derived from food-grade bacteria, are adapted to mucosal delivery of vaccines. According to Mucosis, there are two modes of application of the BLPs. The first is an add & mix approach in which the immune-stimulating properties of the BLPs are used to improve existing vaccines and to enable a mucosal route of administration. The second is a bound approach in which the BLPs not only act as an immunostimulant but also as a carrier for the subunit antigen(s). The binding of the antigens is achieved by a linker domain called Protan that binds to the surface of the BLPs. Mucosis is developing its adjuvant technology in a vaccine pipeline that includes FluGEM, an influenza vaccine using the BLP technology, which is in Phase I clinical trials 7.

Tetragenetics recently announced preclinical study results showing that an investigational nanoparticle-based influenza vaccine using the G-SOME platform protects against a highly pathogenic H5N1 avian influenza strain. The study was conducted by scientists at the Institute for Antiviral Research of Utah State University in Logan, Utah and Tetragenetics, under a contract sponsored by the National Institute of Allergy and Infectious Diseases (NIAID), part of the National Institutes of Health. The recombinant nanoparticle vaccine against a highly pathogenic avian H5N1 (A/Vietnam/1203/2004) strain at lethal doses led to 90% survival following immunisation and lethal challenge. The H5N1 vaccine candidate was comprised of the viral hemagglutinin linked to a scaffold protein from Tetrahymena thermophila that promotes assembly into remarkably homogenous nanoparticles in the 40-60nm size range (G-SOME™ particles6). New Strategies for Antigen Design – Focus on Recombinant Subunits and VLPs Rational design of vaccines has been one of the new areas of development that has given the vaccine field a new interest, with tools that allow the design and manufacturing of what is expected to be more efficient and safer vaccines. New vaccines such as Prevnar (recombinant subunits), Cervarix and Gardasil (VLP) have demonstrated the market potential of new technologies. New approaches that rely on recombinant technologies and that deliver the vaccine either in the shape of recombinant DNA, or the resulting recombinant protein, which in specific cases can assemble in a particle, include particles that mimic viruses: virus like particles, or VLPs. Amongst the new types of vaccines being developed, we will focus here on recombinant subunit vaccines and VLPs. In both cases, the products in development make use of industrial platforms for recombinant protein production. The NIAID gives the following definition for recombinant subunit vaccines: Instead of the entire microbe, subunit vaccines include INTERNATIONAL PHARMACEUTICAL INDUSTRY 39

Drug Discovery/Development & Delivery only the antigens that best stimulate the immune system. In some cases, these vaccines use epitopes—the very specific parts of the antigen that antibodies or T cells recognize and bind to. Because subunit vaccines contain only the essential antigens and not all the other molecules that make up the microbe, the chances of adverse reactions to the vaccine are lower. Subunit vaccines can contain anywhere from 1 to 20 or more antigens, (..) [manufactured] using recombinant DNA technology.8 VLPs mimic the external protein structure of a virus without including the genetic material (DNA or RNA) necessary for viral replication. Without genetic material, VLP vaccines are incapable of causing infections themselves while at the same time presenting viral antigens in the most authentic configuration possible. The objective is that the immune system responds to a VLP vaccine as if to the live virus, without the risk associated with virus replication, hopefully making the technology an efficient and safer advance. A variety of platforms have been used for production of VLPs, such as plants, mammalian and insect cell lines. In all cases, successful production of VLPs has been demonstrated, and its efficacy demonstrated through clinical trials. Recent advances using the VLP technology showed strong immune response for vaccines created from VLPs vs. traditional whole viron vaccines, as well as protein antigenbased vaccine. Further possibilities of VLPs were demonstrated by the recent RTS.S hybrid malaria-hepatitis B VLP, which showed 50% protection of children in Africa in Phase III clincal trials 1. Developments in Manufacturing Process – Single-use Units of Operation Manufacturing used to be – and still is to some extent – one of the most important entry barriers to the area of vaccines, with high initial capital investment required for building the production facilities. The introduction and development of single-use components in all unit operations for manufacturing, and the good match between these and the new platforms 40 INTERNATIONAL PHARMACEUTICAL INDUSTRY

used in the production of the new vaccines, has reduced the size of this barrier. Widespread application of single-use technology is being welcomed by the industry and more widely adopted: in a recent industry survey9, a majority of respondents indicated that biologics currently in the pipeline will likely be made in single-use bioreactors. Flexibility in manufacturing is seen as one of the most important trends in innovation in this area, as illustrated by the recent award by the American Pharmaceutical Manufacturing Magazine to Sartorius Stedim Biotech (SSB) as a 2012 Innovations Honoree for its new FlexMoSys production platform design. The system combines SSB’s broad, single-use and reusable product portfolio and G-Con’s novel cleanroom pods, and thus offers a totally integrated production process platform. It enables GMP-compliant cleanroom space to be created that is faster, more flexible and significantly less expensive to set up and operate than a conventional, large purposebuilt facility. Standard pods of the FlexMoSys cleanroom system can be conveniently tailored to a customer’s specific needs, as the customer can choose from an entire suite of fully integrated bioprocessing equipment like SSB’s FlexAct solutions. The FlexMoSys pods can be easily moved due to installed air bearings underneath each pod. This is important when modular unit operations must be placed in a different sequence or a pod has to be decontaminated10. The trend also lowers considerably another traditional barrier holding off competition: the complexity of the supply chain. More flexible manufacturing, engendering lower capital investment for building factories, lowers the barriers for expanding manufacturing to areas in the world where it is difficult to make a case for capital-intensive investments in inflexible factories, such as the more traditional vaccine manufacturing facilities would require. The advantages of single-use are being well documented, with thorough modeling indicating significant economic advantages, including the need for reduced capital investment, with improved returns, increased

speed and flexibility due to reduced facility complexity, smaller footprint and portability, and improved process control. Single-use allows for greater flexibility in matching bioreactor size to actual need, since there is no need to decide and fix on a steel tank volume. Extensive application of singleuse is particularly well adapted to multiproduct facilities, since there is significant reduction in validation protocols, reduced downtime, faster turnaround, and significant reduction in cleaning and sterilisation between products/strains. Through elimination of need for cleaning in place and steaming in place, it was estimated that up to 50% of the energy costs and 8-20 times less water are needed when single-use is widespread11. Comparability of cell growth and productivity for a variety of platforms has also been demonstrated. Bavarian Nordic uses a proprietary modified Vaccinia Ankara virus as the active ingredient of its products. For their smallpox vaccine, IMVAMUNE, the company collaborated with the US NIH for the development of a safe MVA-based smallpox vaccine. The success in developing this vaccine triggered an increased demand, and the company decided to invest in its own manufacturing facility, to complement its existing agreements in other manufacturing sites. The challenge was to build a facility capable of supplying 40 million doses annually in a relatively short period of time, at the same time as process industrialisation and optimisation. Single-use technology has been widely used during process development, and a decision was taken to continue to use it. This allowed building activities to be decoupled from process industrialisation, one of the requirements in the challenging timeline. The disposable work involved collaboration between the Bavarian Nordic staff, Sartoriu and Stedim (now Sartorius-Stedim), Wave Biotech, certain media suppliers, and the engineering company NNE. MVA-virus are too big to be filtered, so the whole process need to be aseptic from start to vial. Flexibility in the facility was also a requirement, and there was a decision to minimise Summer 2012 Volume 4 Issue 3

open handling. The implementation of single-use ensured minimised risk of cross-contamination and maximum flexibility. Regulatory facilities and auditors were consulted regarding the design of the facility, and they expressed interest in such extensive use of single-use technologies. Whenever a single-use solution was available, it was incorporated in the design. Bavarian Nordic used suppliersâ&#x20AC;&#x2122; validation packages and evaluated and selected single-use products based on a risk analysis approach that takes into account several factors, including how critical the process step is. The experience of Bavarian Nordic can be useful for other companies wanting to build new facilities that extensively integrate single-use. For this project, singleuse allowed Bavarian Nordic to reach its objectives: short project timeline, highly flexible facility, adaptable to future manufacturing needs, and minimised contamination risk12. With the continuing success of single-use systems, there has been some consolidation in the industry, with large corporations acquiring smaller, innovative ones, where new single-use technology is developed. A relatively wide choice of singleuse technologies and bioreactors is available and is being developed. Companies, such as Cellution Biotech 13, are developing bioreactor alternatives that are well adapted to enable flexibility in scaling-up and

Table 3: Summary of status for DenV, JEV and WNV (13, 14, 15, 16, 17, 18, 19)

compatibility with cGMP. Table 2 summarises the current manufacturing landscape in terms of manufacturing platform and inclusion of single-use technology, using as examples VLP or recombinant subunit vaccine production, both strategies being amongst the most recent - and considered the most promising technologies for vaccines. Both VLPs and subunit vaccines in development have been produced in a broad range of platforms, and all of the manufacturing platforms are well

Table 2: Platforms for VLP and recombinant vaccine production

adapted to single-use. This example shows the good fit between the new manufacturing technologies and novel vaccine development. The following cases will show how the choice of the appropriate manufacturing platform enables the development of vaccines against a set of infectious agents, for which there is a strong unmet need, against Dengue Virus (DVE, Japanese Encephalitis Virus and West Nile Virus (WNV). Case Analysis: Three Examples of Developments in Areas of Unmet Need This section will use three viral infectious diseases as examples of how the new technologies are enabling strategies that make use of the most modern technologies to respond to current serious unmet medical needs. The three viruses belong to the same family and they are all insect-borne. Table 3 summarises the main disease characteristics and the specific challenges in developing a vaccine for these indications. Of the three viral infections discussed, the only one for which a human vaccine is available is Japanese Encephalitis Virus. The vaccines marketed are all lifeattenuated viruses. There are no marketed vaccines for humans INTERNATIONAL PHARMACEUTICAL INDUSTRY 41

Drug Discovery/ Development & Delivery Table 4: Examples of production platforms for experimental vaccines for DenV, JEV, WNV

against Dengue fever or West Nile Virus. A vaccine for veterinary use to protect against WNV is approved. There has been a long history of failed attempts to develop a vaccine for Dengue Virus, and the current trials with tetravalent vaccines are drawing much excitement. The chronology of vaccine strategies used for JEV illustrates well the impact of new technologies in allowing development of safer and potentially more efficacious vaccines: from an early approval of a vaccine extracted from animal tissue, to use of recombinant technology and safer manufacturing platforms, such as cultured cells, in the most recently approved vaccines, IXIARO™ and IMOJEV™. Production Platforms In view of the large unmet medical need, there has been a continued effort to find an appropriate vaccine strategy, and we can find in the pipeline for these three indications all of the most recent approaches to rational vaccine design, including DNA-based, recombinant virus, recombinant subunit and VLP vaccines, to cite some. Amongst the platforms for recombinant protein production used in the development pipeline for these vaccines, we find that 42 INTERNATIONAL PHARMACEUTICAL INDUSTRY

insect-based platforms feature in all three cases. Experimental subunit vaccines against JEV produced with the Balulouvirus Expression Vector system BEVS are in research stage, and recombinant subunit vaccines against DV and WNV produced in S2 cells have been tested in Phase I. Over the past 10 years, insectbased platforms (BEVS and S2 cells) have moved from the research labs to larger-scale production. There are now insect-cell-made vaccine components in the market and in various stages of clinical development in a variety of indications. For a recent review of the use of BEVS, see18, and for use of S2 cells, see for example19. The strengths of the S2 cell-based expression platforms can be summarised as: speed of production, allowing fast access to the required proteins, excellent expression capability, scalability, applicability to a variety of process choices, and regulatory friendliness. S2 cells have been used for expression and analysis of a broad range of protein classes. As the system is based on generation of stable production cell lines, it allows for a broader variety of upstream process options, compared to the obligatory batch process of the BEVS. Proteins have been produced in fedbatch and in perfusion using ExpreS 2, the platform developed by ExpreS 2ion

Biotechnologies. Both methods led to high production yields, with perfusion allowing for smaller footprint installations, a factor that may allow for more flexibility in early stages of production19. In looking for more flexible, low capital cost investment alternatives to manufacturing, S2 cells again are an interesting alternative: the ExpreS 2 platform has been tested in Cellution’s single-use bioreactors with success in a malaria vaccine in pre-clinical development19. S2 cells have also been tested for production of monoclonal antibodies in WAVE systems. The results showed that production in the order of 1g/L could be obtained and that the antibodies were functional20. The variety of manufacturing platforms being used in these three indications illustrates well the importance of developing a broad range of protein expression platforms to increase the potential for matching the needs for each protein. In conclusion: rational design of vaccines with improved efficacy and safety profiles makes extensive use of recombinant technology. Amongst the new types of vaccines are recombinant subunit vaccines and VLPs. These technologies have opened new possibilities for developing vaccines for diseases where there are no Summer 2012 Volume 4 Issue 3

Drug Discovery/ Development & Delivery current products licensed, such as Dengue and Malaria. The availability of a broad range of manufacturing platforms has also contributed to the advancement of the field, allowing vaccine developers more options to express challenging proteins. It is very important that the production platform method has good industrialisation potential and compatibility with cGMP. Stable insect cell systems, such as S2-based, are well adapted to these needs. The recognition of the important role adjuvants play has contributed to increased research in this area, but both the science and the regulatory path are current challenges to development. There are nevertheless several experimental vaccines in development making use of new adjuvant platforms. Matching the flexibility of the manufacturing technology with flexibility in the actual factory has been a recent development, with the broader introduction of single-use systems. References 1. w 2. 3. C  arlson, B. Genetic Engineering News, Nov 1, 2011 (Vol. 31, No. 19) Vaccine Market Revenue Growing 4. F  rost and Sullivan: 2011 Oct Industry Insights on Vaccines (Technical Insights) 5. ImVacs Conference, Coffman, Sher, Seder, Immunity 2010, cited in “Paying the Toll: Getting modern adjuvants to make T-cell responses, Kedl, University of Colorado, Denver., August 2012. 6. 7. 8. h ttp:// vaccines/understanding/pages/ typesvaccines.aspx 9. L anger, E. Gen Engineering News Aug 2012 32 10. h t t p : / / w w w . c l e a n r o o m t e c h n o l o g y. c o . u k / n e w s / article_page/Sartorius_Stedim_ named_as_2012_Innovations_ Honoree/79880 11.World Vaccine Congress, Lyon 2011. Levine, H-Vaccine manufacturing capacity for the coming decade,

12. W  assard, K, and Monge, M. BioProcess Int. 5:S32-S38. 2007. Rapid Implementation of a Smallpox Vaccine Facility: A Case Study in Single-Use Technology.. 13. 14. Guy, B., Barrerea, B., Malinowskib, C., Saville, M., Teyssouc, R., Langa, J. Vaccine 29 (2011) 7229– 7241 From research to phase III: Preclinical, industrial and clinical development of the Sanofi Pasteur tetravalent dengue vaccine. 15. Clemens, D. et al. Vaccine 28 (2010) 2705–2715. Development of a recombinant tetravalent dengue virus vaccine: Immunogenicity and efficacy studies in mice and monkeys 16. w ww.exanebnpparibas-equities. com: September 2009, Equity Research: Crucell/Intercell 17. De Filette et al., Veterinary Research 2012, 43:16 Recent progress in West Nile virus diagnosis and vaccination. 18. Lim, S.M., Koraka, P., Osterhaus, A.D.M.E. and Martina, B.E.E. Viruses. 2011 June; 3(6): 811– 828. West Nile Virus: Immunity and Pathogenesis 19. Cox, M.M.J. Vaccine 30 (2012)

Dr Sancha Salgueiro, MSc, PhD, MBA VP Business Development at ExpreS2ion Biotechnologies. Dr Sancha Salgueiro has over 15 years of executive experience in biotechnology, in R&D and global business development. Before joining Expres2ion, she held leading business development positions at companies in Denmark, France and in the US, focusing on executing and driving partnerships with biotech and pharmaceutical companies with the purpose of leveraging the companies’ technical platforms for enabling new drugs and providing alternatives for protein production. She successfully negotiated deals with a number of biotech and pharma companies worldwide. Email:

1759– 1766, Recombinant protein vaccines produced in insect cells 20. D  yring et al. IPI-Volume4-Issue2-2, A Robust, Scalable Platform for Recombinant Protein Expression 21. W  ang, L., Hu, H., Yang, J., Wang, F., Kaisermayer, C. and Zhou, P. Molecular Biotechnology 2011, DOI: 10.1007/s12033-011-9484-5 High Yield of Human Monoclonal Antibody Produced by Stably Transfected Drosophila Schneider 2 Cells in Perfusion Culture Using Wave Bioreactor

Dr Charlotte Dyring, MSc, PhD Co-Founder, CEO at ExpreS2ion Biotechnologies. Dr Dyring has an extensive track record in protein expression in eukaryotic systems, mastering a wide array of expression systems, tools and techniques, and having substantial practical experience with upstream process development according to industry standards, including process transfer to cGMP. Dr Dyring is a recognised world-leading expert on the Drosophila S2 expression technology, and her expertise was critical in the development of the Expres2 technology to the current level of sophistication and robustness. Email:

Dr Wian de Jongh, MSc, PhD CSO and co-Founder at ExpreS2ion Biotechnologies. Dr de Jongh has a BSc and MSc in Chemical Engineering from the University of Stellenbosch in South Africa, and a PhD in Biotechnology from the Technical University in Denmark. Dr de Jongh has been instrumental in developing a Expres2 protein expression system, and has more than six years of experience in applying the system to process development and clinical material manufacture. Email: INTERNATIONAL PHARMACEUTICAL INDUSTRY 43

Drug Discovery/ Development & Delivery

Reduce Costs and Gain Expertise by Outsourcing your Cell Culture and Cryostorage Cell culture has evolved over more than a century into a core technique that is central to a diverse range of research and clinical applications. These range from the study of basic biological processes through to stem cell therapy and tissue engineering. As well as contributing to ethical progress by helping to reduce the use of animals, cell culture provides a number of key scientific benefits. In pharmaceutical research and development, cell culture has an increasing number of cutting-edge roles, including a contribution to reduced attrition rates by allowing sophisticated early-stage safety and efficacy screening. Scientists are able to rapidly assess the potential efficacy of large numbers of compounds in high throughput screening (HTS) assays, as well as using the technique for target validation and to investigate issues arising in toxicology and ADME. In addition to these established applications in an R&D setting, cells now play an increasingly dominant role in the biomanufacture of therapeutics â&#x20AC;&#x201C; the fastest growing area of the cell culture industry. In parallel with these

developments, establishing and maintaining optimal systems for the culture, cryopreservation and storage of a wide variety of cell types has now become a priority. The Challenges of Cryostorage Cryopreservation is essential to maintain a long-term, consistent supply of precious cultures. In the absence of good cell banking practices and cryopreservation procedures, the continuous culture of cell lines leads to an array of problems including genetic drift, loss of key characteristics and an increased probability of microbial contamination. Such a significant role in biopharmaceutical R&D creates a need for specialist expertise in a wide variety of cell culture technologies. Screening applications utilise both normal and diseased cell lines from humans as well as key toxicology species. Increasingly, there is a need for primary cells in order to address specific disease mechanisms. Primary cells are those that have not undergone an immortalisation process. They therefore generally reflect the in vivo

situation more effectively than an immortalised cell line, but as they only have a finite lifespan, expanding the culture to provide sufficient cells for an HTS assay often requires specialist expertise. Every cell type requires an optimised system of culture medium and reagents, and needs some validation prior to use. In many cases, safety assessment departments face constantly changing requirements in the cell types that need to be cultured, as in vivo toxicology studies can raise issues and questions that need to be examined more closely in vitro, using a specific cell type in one or more species. Some studies require the use of specialised 3D human tissue models, which require significant operator training. This constantly changing landscape inevitably brings corresponding challenges in terms of maintaining in-house expertise within budgetary restrictions. As the pharmaceutical industry undergoes major structural changes, with mergers, acquisitions and site consolidations now regular features, the demands of keeping up with constantly changing requirements can become increasingly complex. Key team members may be lost or face significant downtime during relocation and/or the merging of company cultures. Maintaining key operational cell culture activities and the secure storage of cell banks is a critical requirement during this time. Advantages of Outsourcing Support Services Outsourcing selected activities can significantly ease the pressure and a good outsourcing laboratory will work alongside you as a partner, supporting both current and upcoming needs, allowing you to dip in and out of additional support as and when required. You may choose to use an outsourced cell culture laboratory occasionally to enhance existing inhouse expertise when an unusual short-term requirement arises. Or you might go for a more substantial


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Drug Discovery/ Development & Delivery outsourcing option. Key services include the expansion of cells into master and working cell banks, cell line characterisation, genetic profiling, mycoplasma and sterility testing. When required, the cells can be thawed and grown in the outsourced lab and provided to your team in any format as assay-ready cells, or assays can be carried out for you. If you only have a one-off or short-term need for a cell type, rather than optimising the culture methods in-house, consider asking your outsourced lab to expand the cells for you and provide them in a no-fuss, assay-ready format. There is a good chance they will have prior experience with the cell type in question – and utilising existing expertise could lead to beneficial savings in both time and cost. Choosing the Right Partner The flexible array of outsourcing choices opens up a whole new approach to cell culture, changing and growing alongside your evolving business. However, the right choice of partner is critical. A good outsourced laboratory should be willing to invest the time to listen to and support your needs to at least the same standard and thoroughness that you would expect from your own in-house scientists. Trust, approachability and flexibility are all preferable to a rigid structure that does not lend itself to reacting quickly to changing requirements. The presence of the right equipment, volume capacity, turnaround time, quality systems and data security are all crucial factors when selecting your potential partner. You may choose to work with multiple laboratories to gain the best access to specialist expertise. XCellR8 ( is a company that aims to maximise the productivity of cell culture-based research by providing easy access to specialist services. They adopt a unique approach by combining an outsourcing facility for cell banking services and in vitro contract testing with training events and technical support. In effect, the UK-based company in Manchester will either take on your cell culture work for you, or help to empower your own team through training and technical support. XCellR8’s philosophy is to 46 INTERNATIONAL PHARMACEUTICAL INDUSTRY

share its expertise and maximise the benefits, so support is available at whatever occasional level is required even if your day-to-day policy is not to outsource. The company believes that in doing so it is fulfilling a previously unmet need for the biopharmaceutical industry. In today’s climate, flexibility is key, and you need to be able to choose whatever level of external support is best for you and your team. Outsourcing Cryogenic Storage Successful outsourcing of controlled environment storage provides access to state-of-the-art facilities while enabling significant cost reduction due to economy of scale at the outsourced facility. In addition, it can provide peace of mind, whether the goal is long-term storage or an interim solution. During site consolidations following a merger or acquisition for instance, outsourced storage could be the ideal solution. However, if handled badly, any type of business process outsourcing can unsettle your organisation, resulting in lower quality of services. The right outsourced storage organisation will seek to build effective partnerships and respond quickly to specific market requirements, giving guidance on critical aspects including laboratory processing and liaison with regulatory bodies. Choosing a partner that understands your market sector, and the pressures that are exerted on it, will ensure a more efficient process to help motivate your own plans for the future.

In drug development, cost control is a critical component of maintaining a competitive advantage. If you want to free up aspects of your own operations that might not be running efficiently, it may be worth considering the merits of outsourcing some of your more routine services. Cryogenic storage will allow your team to focus on core competencies while enabling instant access to state-of-the-art, secure facilities without the need for capital outlay, additional space, equipment maintenance or additional training and labour costs. When choosing your preferred outsourcing partner (POP) for cryogenic storage, you should question whether the selected company has the following attributes: • A proven track record. • State-of-the-art facilities. • Computer-controlled guarantees for temperature integrity, with constant monitoring. • Full emergency backup, including spare equipment. • Dedicated service teams on call 24/7/365. • High-security environment, including access control systems, motion detectors, smoke detectors and continual CCTV surveillance. • Reliable sample tracking. • Simple but secure access to your sample information and a straightforward process for sample retrieval when required. • A fully documented disaster recovery plan. • Affordability. Summer 2012 Volume 4 Issue 3

Drug Discovery/ Development & Delivery •T  he financial capability to match the continued investment needed for your requirements and assurance that long-term requirements will be met. If the answers to all of these questions are affirmative, you’ve found your perfect POP. The chosen company must also have a comprehensive quality assurance programme with best practice standard operating procedures, as well as systems in place to demonstrate the use of regular internal audits for compliance with appropriate regulatory bodies. The company should be able to provide security, absolute visibility of process, and reliable access to the materials using current compliant procedures. A comprehensive information management system is at the heart of all storage solutions. The inventory management solution should enable the user to record all data associated with the samples, including location and temperature, as well as recording all user-defined information. Every sample handled must be given a unique barcode label and then recorded on the compliant database system. This significantly simplifies handling, tracking and processing. Supplementary information relating to individual samples in terms of movement within the facility, time and duration of storage vessel openings and who accessed the vessel must also be available as part of a comprehensive audit trail. All staff should be fully accountable and provide a service that complies with regulatory requirements. Cells destined for biomanufacturing and therapeutic applications must be handled at all times according to cGMP guidelines. Clearly the cGMP service provided by your POP should always meet legislative obligations. Effective Disaster Backup Commercial organisations are increasingly looking at their approach to disaster recovery. Backup secondary storage is considered to be an effective insurance policy against the various mishaps that are beyond your control and is an alternative to 48 INTERNATIONAL PHARMACEUTICAL INDUSTRY

total outsourcing. With this model, in parallel with the primary in-house storage facility, a set of duplicate samples is retained by a third party, in a secure second geographical location. The purpose of any disaster recovery plan would include the following benefits: • Minimise potential economic loss. • Insure against the loss of products in storage. • Counteract the disruption to stability storage activities. • Reduce critical loss to your company. • Provide an orderly recovery of product. This approach is well worth considering on the basis that it virtually eliminates the risk of a disaster scenario destroying a company’s valuable stability storage samples and causing commercial damage to the organisation. The use of outsourcing for the cryogenic storage of cells is becoming increasingly popular in the biopharmaceutical industry. Vindon Scientific (www.vindon., based in the UK near Rochdale, has significant experience in controlled temperature storage for the pharmaceutical industry, and its state-of-the-art CryoBank provides companies with access to premium facilities and specialist expertise, while helping to reduce their costs. Some organisations still prefer total control of their own storage. However, depending on the company strategy, it can make more sense to outsource cryogenic storage to an experienced entity based elsewhere. Quite clearly, this kind of move needs careful thought in terms of price, quality of service and flexibility of approach. Conclusion As the pharmaceutical industry undergoes major changes, this could be the ideal time to reassess your current facilities and to carry out a cost-benefit analysis of outsourcing cell culture services and / or cryogenic cell storage for your organisation. Outsourcing isn’t right for everyone but it is well worth consideration, bearing in mind that you can pick and choose exactly which activities make

sense for you to outsource. There may be opportunities to reduce costs and avoid significant project delays through rapid access to specialist expertise. Outsourcing can be strategic, costeffective, efficient and productive – all of which are tempting incentives. When handled competently, the results are well worth the initial effort and can add a valuable new dimension to your biopharmaceutical R&D.

Patrick Jackson Patrick Jackson is Business Development Director, Vindon Scientific Limited, he is a qualified engineer, with over 35 years’ experience in senior management and business development and marketing. Patrick has held a number of Board appointments as; Sales Director, Commercial Director, Operations Director and Business Development Director for a diverse range of companies, he has a successful record of developing business systems to sustain growth in highly competitive markets focussing on formulating working relationships with clients and major sub-contractors and negotiating exclusive trading agreements by developing a client and supplier partnership philosophy. Email:

Dr Carol Barker is the co-founder of XCellR8 Ltd with 15 years of cell culture experience. Carol has a BSc (Hons) in Physiology and Pharmacology and a PhD from FRAME Alternatives Laboratory, University of Nottingham (FRAME = Fund for the Replacement of Animals in Medical Research). She has 5 years experience in drug development (Glaxo, Roche), and 8 years business development experience as Manager of European Operations of the cell culture company, Cascade Biologics. Email:

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Straightforward or Challenging? Why It Matters Where You Place DM&PK Studies Medical research has been a major underlying factor leading to the dramatic reduction in overall morbidity and mortality in the 20th century. In his book, The Devil Under the Microscope,1 Thomas Hagar describes the discovery of sulfa drugs and the first observations of the effects of Penicillium fungi on bacteria. These discoveries had an immediate and dramatic impact on alleviating human suffering and are clear examples of what Carl Sagan meant by his often quoted remark, “Science delivers the goods.”2 It’s why we are passionate about what we do. The synthesis of the sulfa drugs, followed closely by the discovery of penicillin, changed everything, and one can say these marked the beginning of modern drug discovery research. The science of drugs, pharmacology, is divided into two distinct, separate but interactive domains: dynamics and kinetics. Pharmacodynamics (toxicodynamics) or PD (TD) is the study of the effects of xenobiotics (drugs; foreign substances) on the body; pharmacokinetics (toxicokinetics) or PK (TK) is the study of the effects of the body on the xenobiotic, or the study of the sojourn of drug molecules through and out of the body. In Goodman & Gilman’s The Pharmacological Basis of Therapeutics, Leslie Benet defines pharmacokinetics as including concentration-time kinetic relationships, chemical reaction kinetics and biotransformation, and the formation of drug metabolites. 3 PK and PD are key components of the scientific discipline of preclinical drug discovery and development, which can be described as a risk assessment process whereby data obtained in preclinical studies are 50 INTERNATIONAL PHARMACEUTICAL INDUSTRY

used to estimate the usefulness of some agent in preventing, curing, or slowing the progression of human diseases. Regulatory agencies require that a preclinical phase of research be conducted before studies in humans may proceed. A preclinical drug registration plan is formulated, and although it will vary according to differences in programs and therapeutic areas, it will always have drug safety and proof-of-efficacy components. Safety data (the study of the relationship of exposure to toxicity) allow for calculating a safety margin in preclinical studies and ultimately, the early estimation of a Therapeutic Index in humans. Simultaneous to safety evaluation, the study of absorption, distribution, metabolism, and excretion (ADME properties) is central to finding new, safe, and effective drugs. The role of a competent preclinical PK programme in modern drug registration is clear: Clinical failure as a result of PK issues in man can be dramatically reduced. The critical issues to assess can be divided into two conceptual areas: 1. Does the compound exhibit druglike pharmacokinetic properties (does it “look like a drug”)? 2. D oes the compound have properties that will cause safety concerns in man? For example, clearance and bioavailability are important considerations in the first instance while toxicity related to ADME, such as drug-drug interactions (DDIs) and metabolism-related idiosyncratic drug reactions are the important considerations in the second instance.4-8 Four fundamental PK questions must be considered:

1. W  hat are the physico-chemical properties of the drug molecule? 2. W  hat are the kinetics of movement of the drug (metabolite) through tissues and fluids (concentrationtime data)? 3. W  hat are the dynamics of interaction of the drug with proteins, nucleic acids, etc., that influence PK? 4. H  ow has the body changed the drug; what is the metabolism of the drug qualitatively and quantitatively? Most preclinical PK studies are straightforward, and include physiochemical measurements, the determination of pharmacokinetic parameters following intravenous and oral administration, and DDI estimation involving cytochrome P450 (CYPs). The commercial availability of human-derived cells, subcellular fractions, and enzymes with greater knowledge of optimum conditions has streamlined the determination of drug interaction with CYPs, transporters, and other proteins. The combination of modern liquid chromatographymass spectrometry (LC-MS) interfaces with tandem quadrupole analysers has greatly simplified the quantitative analysis of drugs in plasma for PK studies. One area that still presents unique challenges is determining the metabolic profile of drug candidates. Related issues include determining the role of drug metabolism and enteroheptic circulation in drug pharmacokinetics, and the appropriateness of preclinical species in assessing the safety of drug metabolites observed in man. Other examples in preclinical and clinical PK research that require greater expertise are “problem” molecules where physio-chemical properties (e.g., low water solubility) complicate the determination of Ki and IC 50 values Summer 2012 Volume 4 Issue 3

Drug Discovery/ Development & Delivery for CYP interactions and protein binding. These situations require an experienced DM&PK scientific staff. One area of DM&PK service that is by definition “non-routine” and not offered by most CROs is full structural characterisation of drug metabolites. For most situations, drug metabolite structure characterisation up to the new drug application (NDA) is partial characterisation. Our scientific staff is experienced with full (definitive) structural characterisation of drug metabolites that is required for significant drug metabolites at the NDA filing. As an example of variability in the “degree of difficulty” from the metabolism perspective, consider tirilazad and diclofenac (Figure 1): • Tirilazad has a complex structure with two distinct domains and exhibits highly complex biotransformation with over 50 metabolites observed in preclinical species and man. • Diclofenac has a simple structure two chlorines that greatly facilitate identification of metabolites by mass spectrometry and exhibits relatively simple biotransformation with aryl-hydroxylation and Phase I conjugation constituting the majority of the biotransformations in preclinical species and man. Characterisation of the major metabolites for these two molecules represents two vastly different degrees of difficulty. Figure 1. The structures of diclofenac

and tirilazad. Another area of non-routine assessment of drug metabolism is the use of radiolabelled compounds to study formation of reactive drug metabolites and the formation of 52 INTERNATIONAL PHARMACEUTICAL INDUSTRY

covalent bonds of reactive metabolites with macromolecules in vivo and in vitro. Assessing the exposure to drug metabolism in preclinical species relative to exposure in man is crucial and requires expertise to accomplish. The FDA has mandated that regulatory documents must clearly demonstrate preclinical safety data for significant drug metabolites to satisfy the Metabolites in Safety Testing (MIST) Guidelines. Drug metabolites that represent 10% or greater exposure relative to the parent drug in man must also demonstrate a similar exposure level in rodent and non-rodent preclinical toxicology species. In other words, if the exposure in preclinical species is not similar to what was observed in man, then separate, additional toxicology studies must be performed for these compounds (drug metabolites), resulting in considerable delay and expense. Thus, MIST guidelines are extremely important. MPI Research DM&PK scientists are highly experienced in the area of biotransformation and the supporting technology of mass spectrometry, including • Techniques to evaluate the risk of exposure to metabolites

quantitatively in accordance with the MIST guidelines •Techniques described in the literature •T  he development of novel approaches One complicating issue in assessing exposure to drug metabolites is that standards usually do not exist. This is further complicated by the fact that in the absence of authentic standards for drug metabolites, quantitative statements cannot be made concerning exposure (the relative response factor for each metabolite is unknown). One way to circumvent this situation is to directly compare the responses observed for parent drug and metabolites observed in the plasma of preclinical species with the responses observed for the parent drug and drug metabolites in the plasma of man. For example, a pooling and samplecombining technique was recently described by the DM&PK group at Merck Research Laboratories.9 In this clever approach that compensates for analytical differences between species, plasma samples are pooled and analysed separately and as a combination. Comparison of plasma samples Summer 2012 Volume 4 Issue 3

Clinical Research


Drug Discovery/ Development & Delivery obtained from preclinical toxicology studies with human plasma samples will assess if exposure to metabolites was similar and, thus, whether the safety of drug metabolites was adequately tested in the preclinical species. MPI Research high-resolution mass spectrometry instrumentation is ideally suited for comparing the exposure of drug metabolites in preclinical species to the exposure in man to get an estimate of risk, so that MIST risk is quickly assessed. Various approaches have been taken to assess exposure to drug metabolites in preclinical species and in man. One complex and expensive approach is to dose humans with trace doses of [14C]-labelled drug in combination with either microdoses or normal doses of non-labelled drug. Historically the analysis is then accomplished using LC and fraction collection combined with accelerator mass spectrometry (AMS) to determine the exposure to drug metabolites. The drawback to this approach is that the analyte is carbon-14 and the method does not provide information about the nature of the metabolites. We have been exploring “better” ways to accurately determine exposure, and recently described a generic high resolution isotope dilution (HRID) MS method employing in vitro metabolism of isotope labelled compounds. We applied this approach to the quantitative analysis of drug metabolites in plasma, using diclofenac and testosterone as test compounds. We observed that HRID with metabolism of an isotope-labelled compound reduces the number of analytical variables considerably, representing a simpler alternative quantitative method compared with traditional approaches. This method may be a useful approach for addressing issues in the MIST guidelines.10 High resolution LC-MS has changed what is possible in qualitative and quantitative analysis for drug metabolic profiles. Our primary instrumentation for addressing metabolism issues is a modern Thermo LTQ Orbitrap Discovery High Resolution Mass Spectrometer. This instrumentation enables us to determine the elemental composition 54 INTERNATIONAL PHARMACEUTICAL INDUSTRY

of drug metabolites, applying the most modern techniques to establish the drug metabolic profile. MPI Research enjoys an excellent reputation in the industry for supporting preclinical safety assessment efforts for drug registration and has contributed to the successful registration of numerous drugs, biologics, and medical devices. The points described herein make it very clear that it does matter where you place your DM&PK studies. MPI Research has the equipment, the facilities, and the experienced, talented staff to address the most difficult of DM&PK issues. References 1. H  agar, T. (2006). The Demon Under the Microscope: From Battlefield Hospitals to Nazi Labs, One Doctor’s Heroic Search for the World’s First Miracle Drug. Random House, NY. 2. D  emon Haunted World. Sagan (1996) Random House, NY. Science may be hard to understand. It may challenge cherished beliefs…….. But one thing you have to say about it: It delivers the goods. 3. B  enet, L. (1996). Goodman & Gilman’s The Pharmacological Basis of Therapeutics. McGrawHill, NY. 4. P  earson, P.G., and Wienkers, L.C. (2009). “Handbook of Drug Metabolism,” Drugs and the Pharmaceutical Sciences 186, 2nd ed. Informa Healthcare, New York. 5. R  enwick, A.G. (2001). Toxicokinetics: Pharmacokinetics in Toxicology. In: Hayes AW, ed. Principles and Methods of Toxicology. 4th ed. Raven Press, New York, 137-192. 6. R  ogge, M.C., and Taft, D.R. (2010). “Preclinical Drug Development,” Drugs and the Pharmaceutical Sciences 187, 2nd ed. Informa Healthcare, New York. 7. B  eck, B.D., Slayton, T.M., Calabrese, E.J., Baldwin, L., and Rudel R. (2001). Metabolism: A Determinant of Toxicity. In: Hayes AW, ed. Principles and Methods of Toxicology. 4th ed. Raven Press, New York, 23-76. 8. R  odrigues, D.A. (2007). Drug-

Drug Interactions. Drugs and the Pharmaceutical Sciences 179, 2nd ed. Informa Healthcare, New York. 9. Ma, S., and Chowdhury, S.K. (2011) Analytical strategies for assessment of human metabolites in preclinical safety testing. Anal. Chem. 83, 5028-5036. 10. V  rbanac, J., Hilgers, A., Dubnicka, T., Shilliday, F.B., Sams, J., Humphries, D., Slauter, R., Hayes, R.N. (2012). Stable-isotope dilution (SID) high-resolution mass spectrometry (HRMS) quantitative analysis of drug metabolites (DMs) in human plasma without DM standards. 60th Annual Conference of American Society for Mass Spectrometry, Vancouver. WP24, 495.

John James “Jim” Vrbanac, Jr., PhD, is Director of ADME and Senior Study Director at MPI Research. He is responsible for overseeing the ADME department by providing leadership for business development, scientific advancements, and high-quality performance of the ADME team, as well as managing the overall planning and conduct of nonclinical investigations. Prior to joining MPI Research, Dr. Vrbanac has held various scientific leadership positions at PharmOptima, Pfizer, Pharmacia, and The Upjohn Company. Dr. Vrbanac was also Assistant Professor, Pharmacology, Medical University of South Carolina and prior to that was a mass spectrometry specialist at the MSU/NIH Mass Spectrometry Facility. He co-founded PharmOptima and Biopolymer Innovations. Dr. Vrbanac holds a PhD in pharmacology and a BS in biochemistry from Michigan State University. He has 30+ years of experience in mass spectrometry, GC-MS, and LC-MS, and was past Chairperson on the Quantitative Mass Spectrometry Interest Group for the American Society for Mass Spectrometry. Email:

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Near-Infrared Fluorescent Methods for Protein Analysis Fluorescent dyes are important labelling tools for life science research. In recent years, a variety of classic protein analysis techniques have been updated with near-infrared (NIR) fluorescence. Detection in the NIR spectrum (~700-900 nm) offers enhanced sensitivity and signal-to-noise ratios, because the intrinsic autofluorescence of blotting membranes, plasticware, animal tissue and chemical compounds is very low in this spectral region. The performance of visible fluorescent dyes (~400-650 nm) is limited by high autofluorescent background in some applications, including Western blotting. Near-infrared fluorophores, such as IRDye ® infrared dyes, enable quantitative multiplex detection for Western blotting, cell-based assays (In-Cell Westerns™), electrophoretic mobility shift assays (EMSA), and other assay formats. Western Blotting For decades, researchers have relied on Western blotting to confirm the presence or absence of target proteins in complex samples. After electrophoretic separation, proteins are transferred to a membrane support and interrogated with antibodies. Traditional Western blot detection uses a primary antibody directed against a target protein and a secondary antibody conjugated with an enzyme reporter (horseradish peroxidase or alkaline phosphatase). Chemiluminescent or colorimetric detection of the enzyme conjugate confirms the presence of the target protein. Some studies also require quantitative information about how much protein is present (Fig. 1; 1). Fluorescent Western blot detection was developed to meet this need2,3. Secondary antibodies are labelled with NIR fluorescent dyes, such as 56 INTERNATIONAL PHARMACEUTICAL INDUSTRY

Figure 1. Upregulation of ß-catenin, c-Myc, and CtBP1 in colorectal carcinomas. A) Quantitative Western blotting shows higher levels of ß-catenin, c-Myc, and CtBP1 in colon cancer cell lines of increasing tumorigenicity (Caco-2 < DLD-1 < HT-29 < HCT116 < SW480), relative to a normal colon cell line (CCD-18Co). Odyssey CL Imager was used. B) Quantification of Western blot data in panel A. All values were normalised to ß-actin levels. Gujral, T.S. and MacBeath, G. PLoS ONE 5(4), e10024 (2010).

IRDye® 800CW or IRDye 680RD, for direct, non-enzymatic detection. Blots are then documented with an NIR laser imager (Odyssey® CLx, Odyssey Fc, or Odyssey Sa Imagers from LICOR ® Biosciences). With this ratiometric approach, fluorescent signal is directly proportional to the amount of target protein present3,4,5. Near-infrared fluorescent detection dramatically improves quantitative accuracy and reproducibility, with excellent sensitivity for detection of endogenous protein levels 3,5. Fluorescent signals are stable indefinitely, so blots can be stored and re-imaged later. Multiplexing of NIR dyes (Fig. 2) allows two targets to be detected simultaneously, using secondary antibodies with spectrally distinct IRDye labels. Multiplex detection makes normalisation against an internal control easier and more accurate, because the controls are detected on the same blot and in the same lanes as the target protein. Quantification and Dynamic Range Quantitative analysis can be very important in life science research, and IRDye fluorescence outperforms other Western blot detection methods. Studies comparing chemiluminescent and IRDye detection methods

Figure 2. Multiplexing of spectrally distinct IRDye fluorophores. A) Two different protein targets can be detected. B) Multiplex phospho-analysis is performed by combining a phospho-specific antibody with a panantibody that recognises the target regardless of its phosphorylation state.

demonstrate that IRDye detection is quantitative over a much broader linear dynamic range than ECL™ detection3,4,5,6. Fluorescent conjugates avoid the enzyme kinetics and substrate availability limitations of chemiluminescence. This produces more consistent, straightforward, and accurate quantification3,4). Biological samples can be challenging for quantification. Because endogenous protein levels span an extremely wide dynamic range (an estimated 4-10 orders of magnitude;7), the ability to image both low-abundance and high-abundance targets is critical. IRDye detection Summer 2012 Volume 4 Issue 3

with the Odyssey family of imagers provides the widest linear dynamic range reported for any Western blot method. Detection of Hsp70 was reported to be linear over 4.3 orders of magnitude (up to 20,000 fold), from 5 pg to 100 ng5. Wide linear range allows all necessary information to be extracted from a single blot or image, without multiple exposures. In carefully controlled experiments, absolute quantification can be performed. Wang et al. 4 examined the stoichiometric relationships between p53 and its negative regulators. Cell numbers in each sample were carefully counted before lysis. Hdm2, HdmX, and p53 were then detected on NIR fluorescent Western blots. Results were normalised against an internal control protein. Cell extracts were compared against dilution curves of purified target proteins of known concentration, allowing the authors to estimate actual numbers of protein molecules per cell. Their results suggest that the stoichiometric balance between Hdm2, HdmX, and p53 is important for regulation of p53 activity. Multiplex Protein Analysis Two-colour fluorescent Westerns are frequently used for quantitative analysis of cell signalling. The second fluorescent channel is used for normalisation, to correct for loading variation and make quantification more accurate. One approach is to multiplex a phospho-specific antibody with an antibody that recognises the unmodified target protein (Fig. 2;8,9,10,11). Phospho-signal is then normalised against the total level of that protein, using the target protein as its own internal control (Fig. 3). This method corrects for changes in the overall level of the target protein and provides maximum accuracy. Alternatively, the second colour can be used to detect a different protein of interest or to normalise against a “housekeeping” protein such as actin or tubulin. Multiplex phosphoanalysis requires high sensitivity in both detection channels. Western blotting is also essential for validation of other assays. In mass spectrometry proteomics studies, Western blots provide confirmation

Figure 3. Stimulation of sensory neuron cultures with the ErbB4 receptor tyrosine kinase. Cells were stimulated with solubilised ErbB4 kinase (B4; extracellular domain). Phosphorylation of AKT and ERK in response to stimulation was measured by multiplex Western blotting of whole cell lysates. a) Ratio of pAKT:AKT was significantly increased (*p<0.05) by ErbB4 treatment. This increase was blocked by pre-incubation with wortmannin (WM), an inhibitor of phosphatidylinositol-3-kinase activity. b) Signalling is pathway-specific and did not activate the MAPK pathway (ratio of pERK:ERK is unchanged; p=0.98). Odyssey CL Imager was used. Canetta, S.E. et al. PLoS ONE 6(9), e25108 (2011).

of the observed changes in protein abundance12. Validation is particularly important for RNAi studies, because mRNA levels do not always accurately reflect protein levels. Figure 4 shows siRNA silencing of Akt1. qRT-PCR analysis indicated near-complete silencing of Akt1 (Fig. 4A). However, Figure 4. Validation of siRNA silencing by qRT-PCR and Western blot. Cells were transfected with Akt1 siRNA or scrambled siRNA. Quantitative real-time PCR (qRTPCR) and quantitative Western analysis were performed.A) After Akt1 siRNA transfection, the Akt1 mRNA transcript was undetectable for >40 cycles of qRT-PCR, indicating nearcomplete silencing. B) In contrast, quantitative Western blotting of those same samples demonstrated incomplete (66%) reduction of Akt1 protein (13). Actin was used as an internal control. Odyssey CL Imager was used.

quantitative Western blotting of the same samples showed incomplete (66%) reduction of Akt1 protein (Fig. 4B;13). Quantitative Western analysis considers the impact of protein stability, yielding a more complete picture of RNA silencing. In-Cell Western Immunofluorescent Assay IRDye fluorescence is also used for cell-based assays, to efficiently quantitate protein levels and signalling events in many samples. The In-Cell Western™ Assay (ICW) is a microplate assay that uses immunofluorescent staining for fast, accurate measurement of protein levels in fixed cultured cells 6,8,10. Fluorescent signals are linear across a wide range of cell densities. InCell Western assays are also called quantitative immunofluorescence assays, cytoblots, cell-based ELISA, In-Cell ELISA, and cLISA. The moderate-throughput ICW assay is based on standard immunofluorescent techniques. Cells are grown in 96- or 384-well microplates, exposed to the desired treatments or conditions, then fixed and permeabilised for immunostaining (Fig. 5). Total fluorescent signal for each well is imaged and ratiometric analysis is performed. Two fluorescent channels can be used to simultaneously quantify two different protein targets, normalise against a housekeeping protein, or normalise for cell number using a DNA stain10,14,15. Normalisation increases accuracy by correcting for unavoidable well-to-well variation in cell number. The ICW assay provides a “snapshot” of the protein expression or signal transduction status of the cell population in each well. It is a convenient alternative to Figure 5. In-Cell Western assay method. Cells are cultured and treated in microplates. After fixation and permeabilisation, multiplex immunofluorescent staining is used to detect and quantify protein targets. Total fluorescent signal from each well is imaged.


Drug Discovery/ Development & Delivery flow cytometry for analysis of adherent cells and is much simpler than high content screening15. In-Cell Western assays are a useful alternative to Western blots. Timeconsuming, error-prone steps such as cell lysis, electrophoresis, and membrane transfer are eliminated. The entire procedure is performed in a microplate, and many samples can easily be processed in parallel. Because sources of variability are reduced, standard deviations are much lower for ICW assays than for Western blots 6,16. Enhanced reproducibility enables more accurate analysis of changes in protein levels. This is especially useful for detailed signal transduction studies that require precise timing and sampling over an extended period (Fig. 6;14), or evaluation of many different treatment Figure 6. Kinetics of cellular ß-catenin accumulation upon stimulation with Wnt3a. ICW assays were used to measure ß-catenin levels in 384-well microplates. A) Time- and dose-dependent accumulation of cellular ß-catenin. L-cells were incubated with Wnt3a, then stained for ß-catenin (yellow in merged image) and DNA content (red). Plates were imaged with Odyssey CL Imager. B) Quantification of ß-catenin accumulation. Levels were upregulated within 30 min of Wnt3a stimulation, exhibited increased intensity between 6–8 h, and started to plateau after 10 h. Graph shows two independent experiments, each done in quadruplicate. Hannoush, R.N. PLoS ONE. 3(10), e3498 (2008).

conditions (Fig. 7). The ICW assay is well-suited for analysis of protein phosphorylation (Fig. 7) and the effects of drug compounds on signalling pathways, including IC50 determination6,8,10,14. The assay has been used for a variety of applications, including monitoring of GPCR functional activity8, screening for inhibitors of Tau protein 17, viral quantification18, caspase-3 activation assays 19, and RNAi library screening15. Modified ICW assays are used to analyse and quantify cell surface proteins, receptor trafficking, and receptor internalisation20,21. Figure 7. Phosphorylation of myosin regulatory light chain (PMLC20). Uterine myocytes were treated with oxytocin (OT; 10-14 to 10-6 M) for 20 sec in the presence of pharmacological inhibitors. PMLC20 was measured by ICW. Response to OT was diminished by inhibitors of MLCK (ML7), CaM (W7), phosphatidyl inositol-PLC (Edelfosine), and L-type Ca2+ channels (Nifedipine). Relative PMLC20 levels (normalised to cell content) are expressed as a percentage of vehicle control. Aguilar, H.N. et al. PLoS ONE 5(4), e9965 (2010).

Protein/Nucleic Acid Binding: Electrophoretic Mobility Shift Assays (EMSA) The EMSA (electrophoretic mobility shift assay) method is used to study protein:DNA complexes and interactions. When electrophoresed on a non-denaturing gel, protein:DNA complexes migrate more slowly than unbound linear DNA (Fig. 8). Because DNA migration is “shifted” when bound to protein, this method is also called a gel shift assay. Short linear DNA fragments that contain consensus binding sequences are typically used as probes. Radiolabelling of probes with 32P is commonly performed. Existing mobility shift assay protocols can be easily converted to NIR fluorescent assays by replacing the 58 INTERNATIONAL PHARMACEUTICAL INDUSTRY

Figure 8. EMSA/gel shift assay. Protein:DNA complexes migrate more slowly than unbound probe, causing an electrophoretic mobility shift. When a large molar excess of unlabelled competitor DNA is added, this mobility shift is greatly reduced. The competitor DNA typically contains exactly the same consensus sequence as the labelled probe. Competitors with mutated binding sites can also be used.

radiolabelled DNA oligonucleotides with IRDye® oligonucleotides that are end-labelled. Binding and electrophoresis conditions are very similar to other EMSA detection methods. After electrophoresis, the protein:DNA complexes are detected. If a radioactive probe is used, the gel is dried and exposed to X-ray film to document the binding results. If an IRDye fluorescent probe is used, the wet gel is imaged and binding results are available immediately22,23,24. An example of NIR fluorescent EMSA analysis is shown in Fig. 9. Figure 9. NF-KB DNA binding activity in tumour tissues. EMSA was used to evaluate NF- KB activation in tumour remnants derived from a xenograft mouse model of pancreatic cancer. Treatment with Difluorinated-Curcumin (CDF) or curcumin (CUR) triggered downregulation of NF- KB activation. Gemcitabine (GEM) activated NF- KB binding. Combined treatment with CDF and GEM decreased NF-KB activity, compared to combination of CUR and GEM. Anti-tumour activity of CDF may involve inactivation of NF- KB. Odyssey CL Imager and NF- KB IRDye 700 consensus oligos were used. Bao, B. et al. PLoS ONE 6(3), e17850 (2011).

Summer 2012 Volume 4 Issue 3

Summary Near-infrared fluorescence is a powerful tool for protein analysis. Autofluorescent background is extremely low in the NIR spectral region. IRDye fluorophores exploit this property, enhancing detection sensitivity and signal-to-noise ratios. Near-infrared fluorescent detection improves the accuracy and performance of Western blotting and other in vitro assays. In addition to the methods discussed here, NIR dyes are used for applications such as biochemical assays, microscopy, tissue section imaging, and in vivo molecular imaging of small animals 25. References 1. Gujral, T.S. and MacBeath, G. PLoS ONE 5(4), e10024 (2010). 2. Schutz-Geschwender, A., Zhang, Y., Holt, T., McDermitt, D., and Olive, D.M. Application Note, LICOR Biosciences, (2004). 3. Gerk, P.M. J Pharmacol Toxicol Methods. 63, 279-82 (2011). 4. Wang, Y.V., Wade, M., Wong, E.T., Li, Y.C., Rodewald, L.W., and Wahl, G.M. Proc Natl Acad Sci USA 104, 12365–12370 (2007). 5. Bromage, E., Carpenter, L., Kaattari, S., and Patterson, M. Mar Ecol Prog Ser. 376, 123–132 (2009). 6. Aguilar, H.N., Zielnik, B., Tracey, C.N., Mitchell, B.F. PLoS ONE 5(4), e9965 (2010). 7. Mitchell, P. Nature Biotechnol. 28, 665-670 (2010). 8. Wong, S.K.F. Anal Biochem. 333, 265 (2004). 9. P unn, A., Levine, M.A., and Grammatopoulos, D.K. Mol Endocrinol. 20, 3179-95 (2006). 10. Chen, H., Kovar, J., Sissons, S., Cox, K., Matter, M., Chadwell, F., Luan, P., Vlahos, C.J., SchutzGeschwender, A. and Olive, D.M. Anal Biochem. 338, 136-42 (2005). 11. Canetta, S.E., Luca, E., Pertot, E., Role, L.W., Talmage, D.A. PLoS ONE 6(9), e25108 (2011). 12. Jenkins, L.M., Mazur, S.J., Rossi, M., Gaidarenko, O., Xu, Y., and Appella, E. Mol Cell Prot. 7, 716727 (2008). 13. Weldon, S., Kovar, J.L., Urlacher, T.,

and Hoffman, G. Poster: Biomolecular Screening Symposium: Advanced Applications Across Academia, Government & Industry (2010). 14. Hannoush, R.N. PLoS One. 3(10), e3498 (2008). 15. Hoffman, G.R., Moerke, N.J., Hsia, M., Shamu, C.E., and Blenis, J. ASSAY and Drug Development Technologies 8(2), 186-199 (2010). 16. Coevoets, R., Arican, S., Hoogeveen-Westerveld, M., Simons, E., van den Ouweland, A., Halley, D., and Nellist, M. Eur J Hum Genet. 17(3), 301–310 (2009). 17. Dickey, C.A., Eriksen, J., Kamal, A., Burrows, F., Kasibhatla, S., Eckman, C.B., Hutton, M., and Petrucelli, L. Curr. Alzheimer Res. 2, 231 (2005). 18. Counihan, N. A., Daniel, L.M., Chojnacki, J., and Anderson, D.A. J. Virol. Methods 133, 62 (2006). 19. Vergote, D., Butler, G.S., Ooms, M., Cox, J.H., Silva, C., Hollenberg, M.D., Jhamandas, J.H., Overall, C.M., and Power, C. Proc. Natl. Acad. Sci. USA 103, 19182 (2006). 20. Hirsch, A., Meimaridou, E., Fernandez-Cancio, M., Pandey, A.V., Clemente, M., Audi, L., Clark, A.J.L., and Flück, C.E. J. Clin. Endocrinol. Metab. 96, E65-72 (2011). 21. Chan, L.F., Webb, T.R., Chung, T.T., Meimaridou, E., Cooray, S.N.,





Guasti, L., Chapple, J.P., Egertova, M., Elphick, M.R., Cheetmas, M.E., Metherell, L.A., and Clark, A.J.L. Proc Natl Acad Sci USA 106, 6146 – 6151 (2009). B  ao, B., Ali, S., Kong, D., Sarkar, S.H., Wang, Z., Banerjee, S., Aboukameel, A., Padhye, S., Philip, P.A., and Sarkar, F.H. PLoS ONE 6(3), e17850 (2011). G  lanzer, J.G., Liu, S., and Oakley, G.G. Bioorg Med Chem. 19(8), 2589-95 (2011). Z  hang, S., Metelev, V., Tabatadze, D., Zamecnik, P., and Bogdanov Jr, A. Proc Natl Acad Sci USA. 105(11), 4156-61 (2008). K  ovar, J.L., Simpson, M.A., SchutzGeschwender, A., and Olive, D.M. Anal Biochem. 367(1), 1-12 (2007).

Amy SchutzGeschwender is a Principal Scientist at LI-COR® Biosciences. She received her PhD in molecular and cellular biology from the University of Colorado at Boulder, and joined LICOR in 1998. After a number of years in R&D, she now focuses on marketing communications and science writing. Email:


Clinical & Medical Research

Paediatric Clinical Trials in Europe and the US: Challenges and Solutions Abstract Pharmaceutical and biotech companies have a regulatory obligation to prepare plans for and conduct studies in paediatric patient populations in both EU countries and the US. This article highlights differences and similarities in the paediatric regulatory compliance strategies and priorities of companies in those regions. Introduction Ensuring that paediatric populations are included in the development plans for all new drugs is the objective of European Medicinal Products for Paediatric Use (EC 1901/2006) in the EU and, in the US, PREA (Pediatric Research Equity Act, which was adopted in 2003 with “sunset” provision, but recently enacted by the US Congress as permanent regulations to be administered by the FDA. Ideally, biopharma companies will conduct this paediactric research in tandem with their adult clinical trials for new pharmaceuticals. At least that is the intent. The reality, according to a study by a team of paediatric investigators, is that there continues to be a disparity between the ratio of research conducted among adults vs. that among children. The authors of that study looked at clinical trials including those which were conducted among patients with the following conditions: asthma, migraine headaches, schizophrenia, depression, diarrhoeal illness, lower respiratory infection, malaria, bipolar disorder, and HIV/AIDS. Sixty per cent of the disease burden of these conditions affects children, and yet only 12% of clinical studies were performed among a paediatric population. To what can we attribute this? Part of the explanation may lie with the geographical spread of disease, along with funding from governments or not-for-profit organisations. Our research indicates that compliance 60 INTERNATIONAL PHARMACEUTICAL INDUSTRY

with paediatric regulations is still a problem for many biopharma companies. Survey of Pharma and Biotech Companies Our team surveyed pharmaceutical and biotech companies to learn what the barriers are to full compliance with paediatric regulations, and to what extent their attitudes may play a part. After identifying the correct decisionmakers in research and development, we asked them to complete an online questionnaire. Respondents shared the following: – Their grasp of regulatory requirements in the US and the EU; – Their thoughts on how those regulations might be improved; – The effects that the regulations have had on their company, especially on their R&D resources; – Their attitudes toward outsourcing paediatric activities; and – The major obstacles they faced in implementing paediatric research. Results We fielded the questionnaire to 55 participants at different companies. Pre-questioning qualified the participants as R&D decision-makers. A majority (61%) of respondents were based in North America. Table 1 and Figure 1 each show the geographical distribution of countries involved in the research, as well as the size of the organisations in the survey. Table 1: Geographical distribution of respondents

Grasp of Paediatric Regulations In Europe, most survey respondents had full or partial understanding of European regulations. Still,

21% indicated they lacked any understanding of the regulations. A further 6% had never heard of PIP. A similar pattern emerged among US respondents, with most having a full or partial understanding of PREA regulations. In fact, 17% indicated they had no understanding of PREA. A further 10% had never heard of the regulations. Impact on R&D Resources Generally speaking, R&D budgets have increased because of the need to prepare for and execute paediatric studies. In fact, 50% of respondents said their budgets had increased to accommodate PREA/PIP commitments. What bears further investigation are the 10% of respondents who felt that their company’s investment in new products has decreased as a result of the regulations. Delivery of Paediatric Activities Identifying an adequate number of patients was overwhelmingly considered to be the biggest challenge. More than 50% said that if all companies submitted PIP/PREA studies the eligible population would be tapped dry. Outsourcing of Paediatric Activities Survey participants were asked how likely they would be to outsource their paediatric clinical trials. Figure 2 below displays those responses; the majority of companies indicated they were either very or highly likely to outsource their activities in either Europe or the US. Companies expressed a desire for a wide range of outsourced support services. Given the previous responses, support in doing patient recruitment was the service in highest demand. Figure 3 below shows the range of requested expertise that would be sought of external partners.

Summer 2012 Volume 4 Issue 3

Clinical & Medical Research Revisions of Paediatric Legislation Contemplated When asked to rate which changes could be made to PREA and PIP legislation, extension of the patient exclusivity period ranked highest. While this was not relevant to PIP studies, the second highest area of importance for PREA and the most important area for PIP regulation was the desire that the rules be simplified — the goal being to improve understanding. Discussion Clearly, the introduction of legislation in Europe and the US mandating paediatric research has resulted in an increase in paediatric research funding. Since 2007, more than 165,000 paediatric patients have been included in completed PREA and BPCA (Best Pharmaceuticals for Children Act) studies. Types of studies are displayed in Table 2 below. Table 2: FDAAA completed paediatric studies between September 2007 and March 2012

Although the regulations have been on the books for many years, it is evident from these results that an unacceptable number of companies are not acting promptly to comply with the rules. While best practices dictate that the compliance process be initiated by companies at the earliest stage of drug development planning, procrastination instead seems to be policy among too many R&D officers. Worse yet, a considerable minority of companies are not yet aware these regulations are in force. The fact that paediatric R&D budgets have increased certainly is good news for the outsourcing industry. But at the same time, companies’ in-house expertise has improved too. Still, there remains a considerable need at biopharma companies for external partners, particularly for partners wellversed in negotiations with regulatory bodies. Companies which have planned their PREA/PIPs compliance still have to deal with the bottleneck of insufficient patients for the studies. In Europe, there are >700 62 INTERNATIONAL PHARMACEUTICAL INDUSTRY

submitted PIPs or waiver applications. The therapeutic areas with the greatest number of PIP submissions are respiratory, endocrine, and CV disease. While some therapy areas (e.g., asthma) will have a high burden of disease in children, other areas will experience recruitment difficulties. For example, the investment in adult oncology research over the past decade has translated to an overemphasis of studies in a paediatric population vs. the incidence of disease. Feedback from our paediatric networks demonstrates that diabetes and renal disease are two research areas that have numerous research programmes nearing saturation. Given such a competitive environment for patients and resources, how can patient recruitment be improved? To start with, as indicated before, best practices dictate early engagement with industry experts as well as investigators. Certainly, the requirements of US and EU regulators differ in a number of ways. What remains constant, however, is that an aligned development programme will reduce the need for study duplication. Working with regulatory and study design experts will weed out simple design flaws that can seriously impair the prospects for recruiting enough participants. Performing a feasibility assessment — even at the PIP/PREA planning stage — will arm companies with the facts about designing a realistic programme before they enter into negotiations with the regulators. While there is yet to be a precedent for what would happen to a company’s market authorisation if it failed to deliver on its obligations, which company would care to be the test case? Increasingly, cooperative nations and groups have joined hands to encourage and facilitate paediatric research. In the UK one such group is Medicines for Children Research Network (MCRN), a government-funded organisation that deals with both industry- and nonindustry-sponsored studies. Working within target timelines, MCRN is wellorganised to support studies at all stages of planning, implementation and research. Paediatric populations are challenging to work with. It would be an understatement to say that children may object to the rigours of clinical trials. Such rigours include recurring medical

visits, physical examinations, invasive tests, and blood draws that may not be necessary in a real-world situation. We have found that tactics which help include scheduling visits after school hours and during school holidays or organising follow-up visits that minimise disruptions to family routine and employment. We cannot overemphasise the importance of educating the family about the clinical trial process once a study is initiated. Actually, participation rates improve when families understand that a clinical trial can have a curative effect on their own child in the study, not to speak of the millions of other children worldwide who are beneficiaries. Summary New regulations concerning drug development have stimulated the design and conduct of paediatric research programmes. While there is still a long way to go before equity is reached with adult research, progress has been substantial. Paediatric programmes that include expert review — especially those which include a negotiation discussion phase with regulatory agencies — can aid in overcoming obstacles in the efficient execution of research studies.

Dr Colin R.W. Hayward, FFPM Head of Medical Affairs, Premier Research. Dr. Hayward is a Fellow of the Faculty of Pharmaceutical Medicine with 13 years international experience in pharmacovigilance, design and implementation of clinical programs and medical affairs. He started his career in industry joining Roche pharmacovigilance and quickly progressing through a series of positions with increasing responsibility to International Medical Leader before joining Prism Ideas as Executive VP in 2007. Colin joined Premier Research in January 2012 and in his current position is responsible for leading the company’s global medical affairs and pharmacovigilance teams. He is a keen advocate of Higher Medical Training for physicians specializing in Pharmaceutical Medicine.

Summer 2012 Volume 4 Issue 3

Clinical & Medical Research

Allergy and Biopharmaceutical Therapy, Part 1: Background and Current Treatment Regimens Introduction A fairly good knowledge base concerning various steps in allergic reactions now exists, but despite this knowledge the prevalence of allergic diseases is still increasing: in some areas of the industrialised world up to 50% of the population is affected. More research is needed to further understand allergic sensitisation and how it can be prevented and treated, and novel biopharmaceutical drugs are needed to increase the treatments available to prescribing physicians and their patients. This paper is the first in a two-part series. It describes the pathological role of immunoglobulin E (IgE) in allergic reactions, and how identification of this role has allowed physicians to treat reactions by addressing the underlying immunological mechanisms. The second paper in the series will discuss unique challenges for clinical trials in this therapeutic area. Prevalence of Allergies An allergy is the body’s immune system response to specific elements in the environment. 1,2 Allergy is characterised by an overreaction of the immune system to a foreign protein, an allergen that somehow finds its way into the body, e.g., being eaten, breathed into the lungs, injected, or touched. This overreaction can result in a wide range of symptoms, e.g., coughing, sneezing, itchy eyes, runny nose, and scratchy throat. In severe cases it can also result in rashes, hives, lower blood pressure, difficulty breathing, asthma attacks, and even death.3 The following information and statistics, presented on the Allergy and Asthma Foundation of America’s website 3 makes the importance of prevention and treatment very clear, since there are no known cures for allergies: • An estimated 60 million Americans 64 INTERNATIONAL PHARMACEUTICAL INDUSTRY

(20%) suffer from some types of allergies: this is approximately twice as many as suffer from diabetes, and many times more than suffer from other high-profile diseases such as Alzheimer’s disease. • Allergy is the fifth leading chronic disease in the US among all ages, and the third most common chronic disease among children under 18 years old. • Approximately 40 million Americans have indoor/outdoor allergies as their primary allergy. Approximately 10 million people are allergic to cat dander, the most common pet allergy. The most common indoor/ outdoor allergy triggers are: tree,

for more than 7 million outpatient visits each year. •F  ood allergies account for 30,000 visits to the emergency room each year. •E  ach year, nearly 400 Americans die due to drug allergies from penicillin, around 200 die due to food allergies, and nearly 100 die due to insect allergies. •A  llergies have a genetic component. If both parents have allergies it is much more likely that their children will have allergies than if only one parent has allergies. Table 1 provides various examples of causes of allergenic physiopathology.

Table 1: Examples of Causes of Allergenic Physiopathology Category Inhalants

Food Occupational sources Pollutants

Examples Mites: Der. Pteronissiums, farinae, … Pollens: Grass (ryegrass, timothy-grass), weeds (ragweed, plantago, nettle, artemisia vulgaris, chenopodium album, sorrel), trees (birch, alder, hazel, hornbeam, aesculus, willow, poplar, platanus, tilia, olea, Ashe juniper), animal danders (cat, dog, rabbit, horse), fungal (moulds, yeasts), insect waste (bee, wasp). Milk, peanuts , tree nuts, fish, crustacea, eggs, soyabeans, sesame, celery and some fruits (apples, peaches…)

Isocianates, flour, grain, resins, metals. Both outdoor (carbon monoxide, nitrogen monoxide or nitric oxide, metals) and indoor (indoor gas pollutants)

Table 2: Examples of Allergy Skin Tests grass and weed pollen; mould Immunoglobulin E Allergens are antigens which induce spores; dust mite and cockroach Category of test Description Prick introducing a small amount of allergen the skin by making a and react with into specific immunoglobulin allergen; and, Involves cat, dog and rodent small puncture through a drop of the allergen extract. Test results are (Ig) antibodies. Antibodies are proteins dander. available within 15 minutes of testing, and the wound heals within 30 • Approximately minutes. 6% of allergy that are used by the immune system Prick-prick have These tests with fresh fruits introduced to reduce poor to were identify, attack, and theneutralise sufferers food/drug allergies of allergy food extracts commerciallyi.e., available. are not in xenobiotics, foreignThey substances as their primarystandardisation allergy. Food standardised and should be restricted to foods for which no recombinant is more common among children the body. These include bacteria and allergen is available. viruses, and neutralisation of these than adults (as are many allergies). Intradermal Involves injecting a small amount of allergen under the skin with a syringe. form reactions of testing is are more sensitive than the prick test method, and substances is a skin desirable outcome. 90% of all foodThis allergy may be used if the prick skin tests are negative. They also attack allergens, but if caused by eight foods: milk, soy, Patch Involves epicutaneous patch tests with allergens known to elicit IgEthis immune system response is too eggs, wheat, mediated peanuts, tree nuts, reactions. Commercial reagents are available for a few allergens. TheyFor have been allergies, standardised regarding use of vehicle dose–response strong,theallergies andand hypersensitivities fish and shellfish. drug relationships. penicillin is the most common can occur. IgE antibodies are found in Scratch Involves abrading the skin and then dropping the allergen on the abraded lungs, allergy trigger. site. (Note: This test shouldtheno human longer be used skin, becauseand of mucous poor reproducibility and possible reactions.) membranes. • Each year, allergies account for systemic Immunoglobulin E (IgE) is one more than 17 million outpatient office visits, which show seasonal subclass of antibodies. Given that influences. Skin allergies account allergic responses typically affect the Summer 2012 Volume 4 Issue 3

skin, gut, and respiratory tract, the major sites of parasitic invasion, it is thought that IgE evolved in humans as a defense against parasitic infestation.3 The IgE test measures the blood level of IgE, one of five subclasses of antibodies. It is often performed as part of an initial screen for allergies. Symptoms of allergies may include hives, itchy eyes or nose, sneezing, nasal congestion, tight throat, and trouble breathing. Some patients suffer from mild symptoms, but others may suffer life-threatening allergic reactions. In general, the quality of life of allergic patients is impaired, although some symptoms may be seasonal (those due to pollen or moulds) or year-long (e.g., mites allergies, animal or food allergies). It is quite unlikely that allergic patients only present one symptom through their life: rather, the “atopic march” is common, as shown in Figure 1. Skin Tests for Identifying Allergen Sensitivity There are several kinds of tests for identifying allergen sensitivity. The rationale is to induce the reaction with the studied allergen. The patient’s medical history will guide which test must be performed. They can be performed in skin, eyes, bronchial regions, and also via food challenges. As an example, Table 2 summarises the different types of skin test. Consider one kind, the skin prick tests. These tests provide an inexpensive, rapid, and accurate method of identifying allergen sensitivity, and they are the most commonly performed tests. The mechanism of action behind their employment is that mast-cell activation causes a characteristic ‘weal and flare.’ There is the possibility of both false positive results (‘identifying’ an allergic response that in truth is not present) and false negative results (failing to identify one when in truth one is present). Therefore, positive and negative controls are always incorporated into the testing procedure. There is a long-standing tradition of extrapolating data from the skin to the airways in clinical practice, largely based on correlations

between skin and upper respiratory effects of antihistamines. But skin tests can be affected by a multitude of factors. These include the quality of the extract, the age of the patient (responses in the elderly are generally less than for other age groups), seasonal variation (skin sensitivity increases after the pollen season and then declines until the next season), and pharmaceutical medicines. Additionally, while a positive skin prick test response identifies sensitisations to a particular allergen, it does not determine the clinical relevance of this sensitisation. On the other hand, a negative skin prick test response has an excellent negative predictive value for excluding IgE reaction. As one last consideration here, the clinical relevance of inhalant allergen sensitisations may differ significantly depending on the allergen.

Correlations between Different Tests Serum-specific IgE, skin prick tests, and allergen challenge do not have the same biological and clinical relevance and are not interchangeable. The weakest correlations have been obtained with mould, food extracts, and non-standardised extracts. There are significant correlations between a strongly positive response to a skin test and the detection of serumspecific IgE, and also between a negative response to a prick test and the lack of detection of serumspecific IgE. However, small weals induced by prick tests and positive results of intradermal tests with concentrated extracts are less frequently associated with the detection of serum-specific IgE. Moreover, low levels of serum-specific IgE are less

Table 1: Examples of Causes of Allergenic Physiopathology Category Inhalants

Food Occupational sources Pollutants

Examples Mites: Der. Pteronissiums, farinae, … Pollens: Grass (ryegrass, timothy-grass), weeds (ragweed, plantago, nettle, artemisia vulgaris, chenopodium album, sorrel), trees (birch, alder, hazel, hornbeam, aesculus, willow, poplar, platanus, tilia, olea, Ashe juniper), animal danders (cat, dog, rabbit, horse), fungal (moulds, yeasts), insect waste (bee, wasp). Milk, peanuts , tree nuts, fish, crustacea, eggs, soyabeans, sesame, celery and some fruits (apples, peaches…)

Isocianates, flour, grain, resins, metals. Both outdoor (carbon monoxide, nitrogen monoxide or nitric oxide, metals) and indoor (indoor gas pollutants)

Table 2: Examples of Allergy Skin Tests Category of test Prick


Intradermal Patch


Description Involves introducing a small amount of allergen into the skin by making a small puncture through a drop of the allergen extract. Test results are available within 15 minutes of testing, and the wound heals within 30 minutes. These tests with fresh fruits were introduced to reduce the poor standardisation of food extracts commercially available. They are not standardised and should be restricted to foods for which no recombinant allergen is available. Involves injecting a small amount of allergen under the skin with a syringe. This form of testing is more sensitive than the prick skin test method, and may be used if the prick skin tests are negative. Involves epicutaneous patch tests with allergens known to elicit IgEmediated reactions. Commercial reagents are available for a few allergens. They have been standardised regarding the use of vehicle and dose–response relationships. Involves abrading the skin and then dropping the allergen on the abraded site. (Note: This test should no longer be used because of poor reproducibility and possible systemic reactions.) INTERNATIONAL PHARMACEUTICAL INDUSTRY 65

Clinical & Medical Research often associated with symptoms than higher levels, but they do not exclude allergic symptoms. Correlations between responses to skin tests or serum-specific IgE and nasal challenges are less consistent because of the non-specific hyperreactivity. Currently Available Therapies Antihistamines have been used for years as a mainstay for the treatment of allergies, including allergic rhinitis.4 There are many different oral antihistamines, which are classified as first generation or second generation depending upon their pharmacologic properties and side-effect profiles. Antihistamines are available as oral, topical and nasal spray. Allergic rhinitis and asthma share a common airway, common mediators, cytokines, and chemokines from mast cells and basophils that are of central importance to the inflammatory response and the series of events that lead to it. 5 As Bachert et al. noted, histamine is the “salient mediator released after immunologic challenge, initiating multiple pathologic processes of the allergic reaction that result in bronchial smooth muscle contraction, vasodilation, mucus hypersecretion, and edema.”5 These authors reviewed 14 clinical trials of second-generation non-sedating antihistamines, and concluded that H1-antihistamines attenuate the symptoms associated with early- and late-phase allergic reactions. Taken together with inhaled anti-inflammatory compounds and bronchodilators, clinical evidence indicates that H1-antihistamines may have a beneficial effect on asthma symptoms and improve quality of life (QoL). Zazzali et al.6 reviewed the cost, utilisation, and treatment regimens associated with chronic idiopathic urticaria. Urticaria is characterised by hives or weals. The term idiopathic indicates that the precise underlying mechanisms of a patient’s chronic condition are not known. They found that antihistamines were the most common treatment, although oral corticosteroids were also commonly prescribed. Therapeutic monoclonal antibodies (mAbs) are increasingly receiving marketing approval. Reichert7 66 INTERNATIONAL PHARMACEUTICAL INDUSTRY

presented data for 34 mAbs that were approved in either Europe or the United States as of March 2012. One of them was omalizumab, a humanized monoclonal antibody that binds circulating IgE antibody. Omalizumab reduces IgE-mediated airway inflammation, and as such is a treatment option for patients with moderate to severe allergic asthma whose asthma is poorly controlled with inhaled corticosteroids and inhaled long-acting ß2 agonist bronchodilators.8 New approvals by the FDA (May 2012) include the combination of antihistamine + corticosteroid as a fixed combination nasal spray. This approval opens a new combination therapy that was not available yet as a single compound, although was commonly used as individual elements together. Allergen-specific immunotherapy is the only approach which may alter the natural course of allergic diseases. Subcutaneous immunotherapy appears to be effective several years after its cessation. During the last decades, allergen-specific immunotherapy was traditionally administered by the subcutaneous route. Now, more convenient oral routes have been developed (oral drops, sublingual administrations). Further Drug Development for Allergy Given the very high prevalence of allergic reactions, as noted previously, there is still a large unmet medical need in this therapeutic area. Further drug development is needed. However, there are unique challenges in the clinical trials required to bring such drugs to market. Part 2 of this mini-series will address these. References 1. World Allergy Organization website. Available at: http://www.worldallergy. org/professional/allergic_diseases_ center/ige/ (Accessed 1st August, 2012) 2. Asthma and Allergy Foundation of America website. Available at: cfm?id=9&sub=19&cont=253 (Accessed 1st August, 2012) 3. Asthma and Allergy Foundation






of America website. Available at: cfm?id=9&sub=30 (Accessed 1st August, 2012) H  oyte FC, Katial RK. Antihistamine therapy in allergic rhinitis. Immunology and Allergy Clinics of North America. 2011;31(3):509-43. B  achert C, Maspero J. Efficacy of second-generation antihistamines in patients with allergic rhinitis and comorbid asthma. Journal of Asthma. 2011;48(9):965-73. Z  azzali JL, Broder MS, Chang E, Chiu MW, Hogan DJ. Cost, utilization, and patterns of medication use associated with chronic idiopathic urticaria. Annals of Allergy, Asthma, and Immunology. 2012;108(2):98102. R  eichert JM. Marketed therapeutic antibodies compendium. MAbs. 2012 May 1;4(3). [Epub ahead of print] T  homson NC, Chaudhuri R. Omalizumab: Clinical use for the management of asthma. Clinical Medical Insights: Circulatory, Respiratory, and Pulmonary Medicine. 2012;6:27-40.

Juan Gispert, MD, is Therapeutic Strategy Lead, Medical Director, and Chair of the Allergy & Respiratory Center of Excellence, Quintiles. He received his MD degree, and then followed a PhD programme in Public Health and Epidemiology, graduating in Design and Statistics and in Pharmaceutical Medicine. He joined pharma in 1992, leading medical and development teams in allergy and respiratory compounds. He has been at Quintiles since 2005. Email: Acknowledgement The author thanks J. Rick Turner, PhD, Quintiles, for medical writing and editing assistance.

Summer 2012 Volume 4 Issue 3

Clinical & Medical Research

Centralised ECGs Help Ensure Cardiac Safety of Obesity Drugs Abstract Obesity rates have dramatically increased over the past 20 years, making it one of the most prevalent health problems in the world. Despite this, there are only a very small number of adequately safe and effective obesity drugs available on the market to treat the condition. Many potential weight-loss drugs have either failed to progress through drug development processes or have been denied approval by regulatory agencies. In addition, many have been withdrawn from the market, having been associated with dangerous cardiovascular events. Due to this, the FDA Endocrinology and Metabolic Drugs Advisory Committee has recently introduced recommendations for drug manufacturers, requiring the submission of Phase II or III clinical trial data demonstrating the absence of cardiovascular risks of new obesity drugs. Centralised ECG trials have emerged as a viable method for helping drug companies provide reliable cardiovascular data in their attempts to achieve regulatory compliance, while also benefiting from significantly improved data accuracy and reliability, time and cost savings, and access to breakthrough technologies. The Obesity Epidemic The latest statistics from the NHS Information Centre for Health and Social Care show that over a quarter of UK adults are classified as obese (26% of both men and women aged 16 and over). 1 In America this figure is even higher, with one-third of US adults classified as obese. 2 In the NHS study, when waist circumference and BMI were used to assess risk of health problems, 22% of men were estimated to be at increased risk; 12% at high risk and 23% at very high risk in 2010. Obesity is a dangerous condition, which substantially increases the risk of morbidity from hypertension, dyslipidemia, type 2 diabetes, 68 INTERNATIONAL PHARMACEUTICAL INDUSTRY

coronary heart disease, stroke, gallbladder disease, osteoarthritis, sleep apnea and respiratory problems, as well as endometrial, breast, prostate and colon cancers. 3 In addition to the high health burden, obesity is also associated with increased medical costs. In a speech given by David Cameron in 2011, it was reported that obesity costs the NHS ÂŁ4 billion a year, and within four years this number is expected to rise to ÂŁ6.3 billion. 4 Safety Issues with Weight-Loss Drugs Until this year, there have been no satisfactorily safe and effective obesity drugs commercially available on the market. Many potential weightloss drugs have either failed or been abandoned by drug development companies and there have been several obesity drugs withdrawn from the market due to safety concerns and adverse side-effects. In 1997, two popular anti-obesity drugs, Fen Phen and Redux, were pulled from the market by the US Food and Drug Administration (FDA) because of potentially life-threatening heart valve damage. 5 Additionally, in 2010, the obesity drug Meridia was recalled for causing an increased risk of stroke and heart attack in populations already prone to these dangerous cardiovascular events. 6 The past few months have seen the approval of two new obesity drugs, the first approvals for drugs of this nature in 13 years. Lorcaserin and Qsymia (formally known as Qnexa) have both been approved by the US FDA recently; however, stringent measures are being put in place to ensure their safety, with them both requiring to undergo a number of post-marketing studies, including a long-term cardiovascular outcomes trial. These therapies are now seeking regulatory approval in additional territories. Another obesity drug, Contrave, is currently seeking regulatory approval again,

after being previously rejected by US regulators over safety concerns. A huge opportunity is presented for pharmaceutical sponsors to make a significant impact on the lives of obese people with long-term drug therapy. However, there have been several challenges, mounting concerns and an increasing number of hurdles required to continue development programmes and ultimately gain regulatory approval for any new weight-loss drugs. In general, weight loss has been tied to positive effects on the heart, but the few trials conducted on the long-term heart safety of obesity drugs have shown that these drugs either increase heart risks, or have little to no effect on heart health. As a result, the FDA will no longer approve a drug for the masses based on a small treatment benefit and without evidence of future improvement on patient health. In clinical trials, Contrave raised pulse rates and blood pressure slightly, a warning sign that the drug might increase the risk of heart attacks, strokes or other cardiovascular problems. As a consequence, in order to gain regulatory approval, the FDA has informed the drug maker that a cardiac safety study must be first conducted to prove that Contrave does not increase the risk of adverse cardiovascular events. 7 Regulatory Requirements On March 29, 2012, after gaining information from experts in obesity, diabetes, cardiology and statistics, the FDA Endocrinology and Metabolic Drugs Advisory Committee recommended that drug companies be required to submit Phase II or III clinical trial data to prove absence of cardiovascular risks for new obesity drugs, even if clinical trials do not initially show evidence of precipitating cardiovascular events. According to the recommendations, obesity trials should randomise approximately Summer 2012 Volume 4 Issue 3

3000 subjects to the active drug and at least 1500 subjects to a placebo for one year of treatment. The FDA requires that patients in Phase III obesity drug trials have a body-mass index (BMI) of at least 30 (or 27 plus comorbidities). The drug should demonstrate either mean efficacy (a >5% difference in weight loss between treatment and placebo) or categorical efficacy (at least 35% of treated patients lose >5% of their baseline body weight, approximately double the proportion in the placebo group). 8 In addition to these already established requirements to prove efficacy, almost all committee members agreed that the major adverse cardiac event (MACE) data should come from either cardiovascular-outcome trials or meta-analyses. Additionally, 17 to 6 voted in favour of the new suggested requirements. 9 Centralised ECG trials can help drug manufacturers comply with the new stringent regulatory requirements, while also enjoying other significant benefits, including dramatically improved data quality and accuracy, time and cost savings, access to cutting-edge technologies and improved overall efficiency of processes. Cost and Time Savings To date, many drug companies have been reluctant to use centralised ECGs due to the common misconception that such an approach is more expensive than a localised or internal model of data collection, primarily as a consequence of the hardware distribution involved. When adopting a decentralised model, the majority of collection, transcription, cleaning and interpretation of ECG data is conducted by the pharmaceutical company and the particular monitoring site. As a consequence, there is a belief that a core laboratory that forms part of a centralised system is a non-essential additional expense. However, a centralised approach is often proven to be more costefficient. A decentralised model requires companies to pay a substantial ECG acquisition fee,

which includes charges for technician time and the use of ECG machines at each investigator site. When using a centralised approach, this fee is reduced since all work is performed in a singe core laboratory. In addition, a decentralised method involves manual transcription, monitoring and quality control of data from multiple sites, increasing labour costs, and data entry errors. Additional cost savings are achieved since a centralised approach dramatically reduces the reconciliation of discrepancies and queries due to inconsistent interpretation during the statistical analysis and medical review phase. In addition, through

implementation of a centralised model, pharmaceutical companies can significantly accelerate analysis time since the collection, transcription, cleaning and interpretation of ECG data are performed using standardised procedures in a singe core laboratory. Here, there is a higher probability to detect cardiac risks earlier in the drug development process, as a result, minimising wasted time and cost expenditure planning for a new drug compound that is not viable. Improved Data Quality and Accuracy When a decentralised model is employed, ECG studies are INTERNATIONAL PHARMACEUTICAL INDUSTRY 69

Clinical & Medical Research conducted across many different investigator sites, using local ECG instrumentation. However, different types of instruments use varying algorithms for calculations, while local investigators or their contracted specialists employ various interpretation guidelines and methods. As a consequence, inconsistent results often occur, presenting a considerable challenge for drug manufacturers, who require accurate and reliable data in order to confidently assess the cardiac safety of compounds in clinical trials. On the contrary, a centralised approach eliminates inconsistencies by facilitating digital collection of high quality data in a standardised format with the use of consistent and validated systems. Qualified individuals assess all interval duration measurements (IDMs), and qualified cardiologists evaluate all ECG data. These cardiologists are trained to follow standardised procedures which are continually validated through a quality control programme. As a result of these standardised processes, companies benefit from much cleaner and more accurate data. Access to Breakthrough Technology In order to gain and maintain a competitive advantage, pharmaceutical companies must continually invest in new, groundbreaking technologies. This is enabled through the adoption of a centralised method, which involves the use of innovative technological solutions, such as lightweight and compact ECG instrumentation. New ECG machines have a significantly smaller footprint and are lighter than their predecessors, thereby removing the challenges raised by the deployment of traditional heavy and expensive instrumentation. Innovative new ECG devices are much easier to manoeuvre and are less costly to ship and store. Due to their compact size, the instruments also offer an improved service on a technical level, enabling more consistency and improved accuracy while effortlessly integrating with computer systems through a simple web application. 70 INTERNATIONAL PHARMACEUTICAL INDUSTRY

Conclusion According to statistics from the NHS Information Centre for Health and Social Care, obesity has more than doubled over the past 20 years in Australia and New Zealand, and increased by half in the United Kingdom and United States. Obesity rates in many Western European countries have also increased substantially over the past decade, making this a global concern. This is an extremely alarming situation since obesity has long been associated with life-threatening health conditions, including cardiovascular diseases. On top of that, many potential weightloss drugs have been found to also cause dangerous cardiovascular events. To address this problem, the FDA Endocrinology and Metabolic Drugs Advisory Committee recommends that pharmaceutical manufacturers should submit Phase II or III clinical trial data demonstrating the cardiac safety of new obesity drugs. Centralised ECG data collection, monitoring and analysis performed in a single core laboratory facilitates regulatory compliance, while also allowing for improved data accuracy and reliability, time and cost savings, and access to innovative technologies. References 1. S tatistics on obesity, physical activity and diet: England 2012. The Health and Social Care Information Centre. webfiles/publications/003_Health_ Lifestyles/OPAD12/Statistics_on_ Obesity_Physical_Activity_and_ Diet_England_2012.pdf 2. P revalence of Obesity in the United States, 2009–2010, Cynthia L. Ogden, Ph.D.; Margaret D. Carroll, M.S.P.H.; Brian K. Kit, M.D., M.P.H.; and Katherine M. Flegal, Ph.D. h t t p : / / w w w. c d c . g o v / n c h s / d a t a / databriefs/db82.pdf 3. N HLBI Obesity Education Initiative, Clinical Guidelines on the Identification, Evaluation, and Treatment of Overweight and Obesity in Adults, http://www. ob_gdlns.pdf 4. H ow much does obesity cost the NHS? May 2001, David Balls:





9. NHS_reforms_David_Cameron_ speech_obesity_costs_foresight_ Department_of_Health-2732 U S Food and Drug Administration, Questions and Answers about Withdrawal of Fenfluramine (Pondimin) and Dexfenfluramine ( R e d u x ) , w w w. f d a . g o v / D r u g s / DrugSafety/Postmarket D r u g S a f e t y InformationforPatients andProviders/ucm180078.htm U S Food and Drug Administration, Meridia (sibutramine): Market Withdrawal Due to Risk of Serious Cardiovascular Events, http:// safetyinformation/safetyalerts forhumanmedicalproducts/ ucm228830.htm N ew York Times, FDA Declines to Approve Diet Drug, http:// w w w. n y t i m e s . c o m / 2 0 1 1 / 0 2 / 0 2 / business/02drug.html F DA committee members grapple with cardiovascular risks in obesity drugs, article/ F DA advisors: “Safe” obesity drugs need to prove absence of cardiovascular risk, http://www.

Robert B Kleiman is a board certified cardiologist and cardiac electrophysiologist who has performed research in both basic cellular electrophysiology as well as clinical electrophysiology. Dr Kleiman completed his training at the University of Pennsylvania, and was a member of a busy cardiology practice for 12 years, before joining ERT full time in 2003. Dr. Kleiman is currently ERT’s Chief Medical Officer and Vice President, Global Cardiology. His responsibilities include oversight of ERT’s cardiology services as well as consulting with external clients. Email:

Summer 2012 Volume 4 Issue 3


Abbes Kacimi, Cold Chain Expert Engineer, SOFRIGAM. Abbes Kacimi is an Engineer in Refrigeration and Air-Conditioning. After a role as the head of Sofrigam Engineering Department (2001), responsible for the development and qualification of cooling containers for pharmaceutical products, Abbes is now the Sofrigam Cold chain Expert Director (2012). He is also member of cold chain commissions (as lIR Commission D2), and Co-author of â&#x20AC;&#x153;Practical Guidelines, Cold Chain for Medicinesâ&#x20AC;?. He contributed to several projects in Europe and Emirates Arabic, and has published a variety of articles on the cold chain. Email:


Clinical & Medical Research

Clinically-relevant in vitro and in vivo Oncology Models for Drug Discovery Introduction The success rate in predicting clinical efficacy of anti-cancer modalities using current xenograft models has been reported to be only 30-40% 1,2. Standard xenograft models use cell lines that are maintained in plastic and have adapted to grow independently of the tumour microenvironment, resulting in models with genetic and phenotypic characteristics distinct from those seen in the clinic 3. In an attempt to reduce drug attrition and improve clinical predictivity, patientderived xenograft tumours (PDX) are being used to improve and refine preclinical modelling. These models provide a more relevant heterogeneous system, in which human tumour and stromal cells are in close co-operation within a unique microenvironment, thereby testing a candidate’s therapeutic agent in the most relevant manner. Maintaining the human microenvironment in models also sustains molecular, genetic and histological heterogeneity of the original tumours, which are important features to ensure authentic responses to current targeted agents or chemotherapeutics. This article will describe how patient-derived preclinical in vitro and in vivo models can be established, characterised and used to enhance the drug discovery process. PDX Models in Drug Discovery Within oncology drug discovery today there are major challenges in identifying novel efficacious and tolerable targeted small molecule and antibody agents suitable for clinical evaluation. To address this challenge, pharmaceutical and biotechnology companies are seeking more relevant and accurate preclinical models to underpin their drug discovery work. The benefits 72 INTERNATIONAL PHARMACEUTICAL INDUSTRY

offered by patient-derived tumour models reside predominantly in their ability to replicate a more authentic tumour microenvironment; this has significant appeal to companies at the forefront of cutting edge drug discovery and oncology research. Such models allow researchers to obtain deeper and more accurate molecular, genetic, SNP, epigenetic and proteomic knowledge relevant to the clinic in order to develop a greater understanding of the disease state. By providing a more clinically relevant heterogeneous microenvironment, it is anticipated that these PDX models will be superior for the discovery and characterisation of novel therapeutics, biomarkers and translational research readouts such as responder/non-responder profiles which can be tested further in the clinic. The key to these models being relevant and useful as a preclinical tool is that a PDX panel for a chosen cancer type should encompass all of the relevant histological and genetic features which make up very specific subsets. It is becoming increasingly clear that the response in the clinic of many targeted agents and some chemotherapeutics is critically dependent upon using these only in the correct tumour subset. For example, lung cancer, which is an area of high unmet need and claims approximately 160,000 deaths/year in the USA, and has a poor five-year survival rate, is composed of several defined histological (especially adenocarcinoma and squamous cell carcinoma) and mutational subsets 4. Over the past decade, it has also become evident that subsets of non small cell lung cancer (NSCLC) can be defined at the molecular level by recurrent ‘driver’ mutations/fusions that occur in multiple oncogenes,

including EGFR, KRAS, ALK, FGFR, BRAF, HER2, AKT1, MEK1, MET, NRAS, PIK3CA, KIF5B-RET and ROS1. For many of these, there are targeted agents, either already registered or currently undergoing clinical evaluation. In some clinical settings, for example the mutant EGFR and EML4Alk subsets of adenocarcinoma (ADC) NSCLC, significant benefits in terms of progression-free and overall survival have been seen in patients treated with targeted agents. This is a fast-growing area with new targets being identified e.g. recently, researchers using wholetranscriptome sequencing identified in-frame fusion transcripts of KIF5B (the kinesin family 5B gene) and the RET oncogene, which were present in 1–2% of lung adenocarcinomas from people in Japan and the US 5. At present, there are several therapeutic agents that include potent inhibition of RET kinase in their profile and warrant clinical evaluation. In addition, lung squamous cell carcinoma (SCC), just like adenocarcinoma, is now starting to split into defined driver subsets, and there is currently major interest in deriving and evaluating targeted agents against the FGFR (mutations and amplifications) and DDR2 (mutations) subsets. A new era in lung cancer is emerging, R&D where defined genetic and histological patient subsets will be selected and evaluated with new targeted agents. It is therefore crucial that representative preclinical in vitro and in vivo models for these defined lung cancer subsets are used to discover, optimise and evaluate new targeted agents and biomarkers suitable for clinical testing. Patientderived lung tumour models are used to establish unique histological and mutational models representing Summer 2012 Volume 4 Issue 3

Clinical & Medical Research the key subsets of interest for drug discovery. For example, adenocarcinoma/mutant EGFR, adenocarcinoma/mutant KRAS, and squamous/mutant/amplified FGFR isolates, are all models currently running within ongoing in vitro and in vivo studies. Establishment of PDX Models In addition to lung models, patientderived xenografts from numerous different cancer types, including colorectal, pancreas, oesophageal, gastric and liver metastases, have been established, which are used for therapeutic evaluation; further expansion of this PDX portfolio will include examples of prostate, breast, haematopoietic and brain tumour models. All samples are collected from the clinic with informed consent following ethical review and are then stored and used for research purposes under the Human Tissue Authority (HTA) license. Associated anonymised patient information, treatment history and histopathological reports are also available. Tracking the patient postsurgery, especially when treatment is pursued, is also critical for validating models as well as reporting back to the clinic on potential secondline treatment options for a specific patient. Tumour samples are implanted subcutaneously in immunocompromised mice, which allows monitoring of tumour establishment and growth by calliper measurement. Typically a lag time precedes exponential growth and this lag time is generally longer in earlier passages, which is likely due to replacement of human stroma by murine stroma. There are also differences in successful establishment across tumour types, and in some cases subsets, as well as dependency on the strain of mouse used. For example, prostate samples are more fastidious than colorectal samples, SCC NSCLC are slightly more amenable to transplantation than ADC NSCLC. Other sites of implantation which improve establishment include orthotopic sites relevant to the cancer type or niche environments such as 74 INTERNATIONAL PHARMACEUTICAL INDUSTRY

the kidney capsule or testes, which are rich in vasculature. In addition, supplementation with matrigel and human stromal cells, such as mesenchymal stem cells (MSCs) or cancer associated fibroblasts (CAFs), which supports retention of the human stroma, may improve tumour take rate. For some models where the paracrine signalling is paramount, stromal components are selected to optimally model the targeted pathway; for example the c-Met/HGF (receptor/ligand) axis is supported through supplementation with MRC5 stromal cells 6. Serial propagation of transplanted tumour material in immunocompromised mice allows sufficient material to be available for therapeutic efficacy evaluation at subsequent passages. Depending on the rate of implant of the tumour, this approach could generate rapid and more clinically relevant data for different drugs and different combinations in a broad set of tumour types and subsets which represent different clinical settings. For this there must be a high correlation between the efficacy readouts of these preclinical models and the clinic to allow accurate predictions of novel therapeutic strategies. Increasing evidence to support the observations that patient-derived tumours in mice can predict the clinical response have been demonstrated in a number of different laboratories 7,8. Establishment of tumour growth rate, stromal retention, doubling rate and generation of donor material between passages are all important factors to catalogue. In addition histopathological characterisation, genetic profiling and response to standards of

care are all key parameters that should be ascertained as soon as tumour establishment has been confirmed. Subset panels can then be clearly identified and used in the most informative way to select for appropriate targeted therapies or strategies for combination. For NSCLC we have identified a panel of SCC and ADC examples and identified responder and nonresponder subsets to a panel of standard of care treatments such as paclitaxel/carboplatin & Iressa. Within each of these subsets there are specific mutations or amplifications that are valuable for therapeutic assessment and understanding resistant mechanisms. A mutant EGFR adenocarcinoma PDX model (Figure 1) harbouring the EGFR mutation L858R shows exquisite sensitivity to EGFR inhibitors such as Tarceva and Iressa, whereas an FGFR amplified SCC model has shown sensitivity to FGFR inhibitors. Other mutations identified include K-Ras, p53 and LKB1. Other areas where patientderived tumour models have added value include: (a) the derivation of chemoresistant isolates to accurately mirror the clinical situation in which new agents are evaluated, (b) to fast-track resistance to new targeted agents and analyse the mechanisms involved and (c) rationally design and evaluate in a high-throughput manner combination studies to delay/ overcome the resistance issues and generate evidence-based hypotheses suitable for testing in clinical trials. Implantation of PDX models at orthotopic sites e.g. intra-thoracic lung injection of PDX cells for NSCLC models (Figure 2), implantation of breast tissue into the mammary fat

Figure 1: ADC NSCLC PDX model with activating EGFR mutation showing exquisite sensitivity to the EGFR inhibitor Erlotinib (Tarceva)

Summer 2012 Volume 4 Issue 3

Clinical & Medical Research pad or direct injection of models into the prostate. These sites provide the necessary microenvironment to promote metastasis, epithelial:mesenchymal transition (EMT) and chemoresistance. Three-dimensional Tumour Growth Assay (3D-TGA) Establishment of a well-characterised panel of tumour types is timeconsuming and costly and together with limited access to original tumour material presents some challenges to deriving PDX models. In vitro establishment of cell lines derived from patient tumour material is very poor, and introducing cells to plastic presents a number of disadvantages to this approach. As an alternative, a 3D-TGA can be established using patient-derived tumour samples which have been disaggregated and admixed with stroma and matrix membrane in 3D to recapitulate the tumour microenvironment as closely as possible in 96- or 384well format. This is in contrast to the evaluation using monolayer single cell cultures, which are standards Figure 2: Intra-thoracic implantation of NSCLC PDX

within the pharma and biotech industries, and whilst predictive to some degree of the biology and mechanism-of-action, are poorly predictive of clinical efficacy. The 3DTGA allows for a rapid non-invasive in vitro measurement of cancer cell expansion in the presence of multiple tumour-associated cell types or soluble factors and facilitates the medium-throughput screening of test agents. The main advantages of the 3D-TGA include (a) the fact that experimental tumour cultures are established in complex mixtures of tumour-derived factors and (b) in comparison to traditional 2D tissue culture systems, both physical and soluble matrix interactions are permitted and spatial limitations are greatly reduced, and therefore in vitro 3D tumour cell cultures are able to more accurately reflect the complex in vivo microenvironment, thus providing a more relevant screening system. Proprietary TGAs can be used to enable detailed pharmacological mapping of multiple agents in isolation, or importantly in

combinations, in clinically-relevant tumour models. Thus, novel compounds in 3D-TGAs, with or without human stroma, are assessed against multiple standards of care, targeted agents or in combination therapies, and can be used to benchmark against competitor agents. The aim is to develop an informative response â&#x20AC;&#x153;heat-mapâ&#x20AC;? of the lead therapeutic agent against and/or in combination with the landscape of existing treatment regimens and develop rational combination strategies in resistant profiles. This allows screening and identification of a more focussed set of agents/combinations suitable for in vivo and ultimately clinical evaluation. This combination approach can also be used to evaluate means to delay or overcome resistance that arises against the targeted agents in specific PDX subsets. Optical Imaging Innovative new approaches are being applied to these PDX in vitro and in vivo models. Through the development of bio-imaging techniques it is possible to monitor in real time the individual tumour and stroma components of the complex microenvironment and how this responds to therapeutic agents. This would allow optimal timing of drug administration to be determined based on the microenvironmental signals measured. This new approach is facilitated by the development of innovative new technology which involves bioluminescent/fluorescent biological reporters. These are engineered to be expressed in human cancer cells so that they emit light or fluorescence in a constitutive manner. Optical imaging also enables monitoring of internal tumours transplanted in mice e.g. orthotopic site such as the lung for NSCLC models or disease progression and metastases. The challenge is to ensure transduction of patient-derived tumour material has minimal impact on the phenotypic and genetic properties of the model. Transduction of the stroma can be achieved through culturing of cells in vitro, however, the patient-derived material may be limited in supply or susceptibility to transduction. As INTERNATIONAL PHARMACEUTICAL INDUSTRY 75

Clinical & Medical Research an alternative, fluorescent-tagged probes can be used to assess internal tumours and also measure a number of different biological responses. For example a caspase-cleavable probe can be used to measure apoptosis in an EGFR mutant NSCLC in vivo model following treatment with Erlotinib and the fluorescent signal can be captured using the Spectrum/ CT imaging system. Concluding Remarks Pre-clinical models that closely recapitulate the human tumour heterogeneity are imperative in the optimisation of novel cancer therapeutics, strategies and patient selection. A comprehensive collection of well-characterised and subsetspecific clinically-derived tumour models provides a powerful screening platform to support the rapid pace of novel target identification/validation, biomarker and drug evaluation studies. In addition these models can be used to better understand the tumour microenvironment, EMT, resistance and metastases and through innovative approaches, such as supplementation with human stroma, optical imaging and 3D modelling, allow these models to be more efficient for oncology drug discovery. References 1. J ohnson JI, Decker S, Zaharevitz D, Rubinstein LV, Venditti JM, Schepartz S, Kalyandrug S, Christian M, Arbuck S, Hollingshead M, and Sausville EA. Relationships between drug activity in NCI preclinical in vitro and in vivo models and early clinical trials. Br J Cancer. 2001 May; 84(10): 1424–1431. 2. F ricker J. Time for reform in the drug-development process. The Lancet Oncology Volume 9, Issue 12, December 2008, Pages 1125– 1126. 3. G  iovanella BC, Stehlin JS, Wall ME, Wani MC, Nicholas AW, Liu LF, Silber R, Potmesil M. DNA topoisomerase I-targeted chemotherapy of human colon cancer in xenografts. Science 1989, Vol. 246 no. 4933 pp. 10461048. 4. C ancer Facts & Figures 2012, 76 INTERNATIONAL PHARMACEUTICAL INDUSTRY

American Cancer Society. 5. K  ohno T, Ichikawa H, Totoki Y, Yasuda K, Hiramoto M, Nammo T, Sakamoto H, Tsuta K, Furuta K, Shimada Y, Iwakawa R, Ogiwara H, Oike T, Enari M, Schetter AJ, Okayama H, Haugen A, Skaug V, Chiku S, Yamanaka I, Arai Y, Watanabe S, Sekine I, Ogawa S, Harris CC, Tsuda H, Yoshida T, Yokota J & Shibata T. KIF5B-RET fusions in lung adenocarcinoma. Nature Medicine 18, 375–377 2012. 6. K umari R, Argent R, Onion D, Clarke PA, King P, Page M and Watson SA. Humanisation of in vivo models to optimally assess the c-met:HGF paracrine axis. EORTC-NCI-AACR Molecular targets and cancer therapeutics meeting, Berlin 2010, poster 272. 7. H idalgo M, Bruckheimer E, Rajeshkumar NV, Garrido-Laguna I, De Oliveira E, Rubio-Viqueira B,

Strawn S, Wick MJ, Martell J and Sidransky D. A Pilot Clinical Study of Treatment Guided by Personalized Tumorgrafts in Patients with Advanced Cancer. Mol Cancer Ther August 2011 10:1311-1316. 8. B ertotti A, Migliardi G, Galimi F, Sassi F, Torti D, Isella C, Corà D, Di Nicolantonio F, Buscarino M, Petti C, Ribero D, Russolillo N, Muratore A, Massucco P, Pisacane A, Molinaro L, Valtorta E, SartoreBianchi A, Risio M, Capussotti L, Gambacorta M, Siena S, Medico

Martin Page As a Scientific Consultant to PRECOS, Martin brings 28 years of experience in Oncology R&D leadership gained from the pharmaceutical and biotechnology sectors across UK, Europe and USA. Martin has led multiple teams discovering novel targeted small molecule and MAb anticancer agents which have entered clinical trials. Prior to his new role, Martin was Vice President, Head of Global Oncology Research for Johnson & Johnson Pharmaceuticals, responsible for laboratories in Belgium and USA. Previously Martin also held senior positions as Oncology Therapeutic Area Head/Europe for Glaxo-Wellcome and Senior Director positions in the biotechnology companies Oxford GlycoSciences and Oncogene Sciences Inc. Martin provides invaluable support to PRECOS in Oncology model development studies for the evaluation of targeted therapeutic agents, biomarkers, translational research and cancer biology.

Dr. Rajendra Kumari, Chief Operating Officer & Founder, PRECOS Ltd. Rajendra received her Ph.D. in Molecular Pharmacology from the University of Leicester in 2002. After a postdoctoral fellowship in the Division of Pre-clinical Oncology, University of Nottingham (UoN), Rajendra took the role of Project & Business Manager of the PRECOS Business Unit where she built up and managed the portfolio of preclinical projects, team and clients, as well as marketing and developing the commercial processes of PRECOS. Rajendra’s expertise in cancer cell biology & model development helped to build PRECOS scientific strengths as well as an academic career as a lecturer at UoN. She is also co-founder of the Ex Vivo Pharmacology Centre of Excellence (UoN), which recapitulates the tumour microenvironment through use of clinically-derived tissue. Rajendra was pivotal in the growth of the business unit, commercialisation of research and the spinout of PRECOS in 2010. Email:

E, Sapino A, Marsoni S, Comoglio PM, Bardelli A and Trusolino L. A molecularly annotated platform of patient-derived xenografts (‘xenopatients’) identifies HER2 as an effective therapeutic target in cetuximab-resistant colorectal cancer. Cancer Discovery 2011, 1(6); 508-523.

Summer 2012 Volume 4 Issue 3

Clinical & Medical Research

Action Duchenne – skipDuchenne Campaign Nick Catlin, Founder and Head of Research at charity Action Duchenne, gives an introduction to the charity, the condition, how they aim to improve life for those living with Duchenne, and the important role that biotech, pharmaceutical and drug discovery companies have to play. Established in 2001, Action Duchenne aims to support and promote innovative research into a cure and effective medicines for Duchenne/ Becker Muscular Dystrophy. The charity, which is led by Duchenne families, aims to promote awareness of the condition, to improve care services, and to provide access to a range of educational and support/ development programmes for people living with Duchenne at every stage of the condition. This is achieved by working in partnership with government agencies, NHS and care organisations, other charities, academic, scientific and research groups, and biotech/pharmaceutical/ drug discovery companies worldwide. Duchenne Muscular Dystrophy (DMD) is an incurable muscle-wasting disease affecting 1 in every 3500 male births in the UK. Duchenne is the most common and severe form of muscular dystrophy, and is the most common genetic childhood killer disease. It is caused by a genetic variation in the dystrophin gene. In every town and every city in the UK there will be at least one boy or young man living with Duchenne; get care and research right for Duchenne and you’ve got it right for thousands of others with related conditions. Since 2003 Action Duchenne has provided £4m for research projects and partnerships. The charity has worked with the MDEX consortium, Department of Health, and the Medical Research Council to deliver new clinical trials for Duchenne drugs. In addition, it has been instrumental in developing projects with biotech 78 INTERNATIONAL PHARMACEUTICAL INDUSTRY

companies both in the UK and US, including key projects with Sarepta (previously known as AVI Biopharma) and Summit. skipDuchenne There are high hopes for the upcoming market approval of small drugs that are able to provide a genetic treatment for DMD by introducing a functional dystrophin protein into muscle cells. Leading the field in terms of advanced clinical trials are antisense oligomers developed by GSK, Prosensa and Sarepta and with a novel drug Ataluren developed by PTC Therapeutics. These molecules aim to trick the RNA splicing mechanism to produce a functional dystrophin protein. Action Duchenne’s skipDuchenne campaign (www.actionduchenne. org/skipduchenne) and funding programme will aim to ensure that the 83% of patients in the UK with DMD that can benefit will have immediate access to exon skipping drugs within the next three to five years. For those young people who are unable to benefit from antisense oligomers (AO) >20% skipDuchenne proposes an alternative strategy using gene replacement delivered via vectors. DMD is caused by mutations in the dystrophin gene that leads to a failure to produce a functional muscle protein called dystrophin. Dystrophin acts as a “coat hanger” for a number of proteins in a complex that provides stability and healthy cells. Cell instability and death will occur when this complex of proteins is misplaced or missing.

Exon Skipping There have been many laboratory studies in DMD animal models over the last 10 years that have shown that by using small molecules called antisense oligomers (AOs) that target specific regions of the faulty gene, a shortened dystrophin protein can be expressed in muscle cells. In the related condition Becker Muscular Dystrophy such a shortened protein can restore the dystrophin complex and muscle function to near normal levels in many cases.

Human studies are now underway targeting the exon 51 region of the gene, and have been shown to be safe and also to successfully express a shorter dystrophin protein.

GlaxoSmithKline and Prosensa are currently conducting extensive Phase IIb and Phase III trials for exon 51 in ambulant and non-ambulant patients and the full data set could be available for 2014. Prosensa has been developing further studies in other exons: Exon 44 Phase I study, Exon 45 and 53 to start in the next wave, 52 and 55 in another wave. Sarepta is continuing with exon 51 human trials in the USA with an alternative AO chemistry that will build upon encouraging data Summer 2012 Volume 4 Issue 3

Clinical & Medical Research already published by the UK MDEX consortium Sarepta also has under consideration other exons 53, 45, 44 and 50. Further studies are planned with the new international exon skipping consortium It is therefore conceivable that if all continues to go well, within the next two years there will be sufficient data available to bring the first genetic treatment for Duchenne Muscular Dystrophy using exon skipping technology to the market. Antisense oligomers can potentially restore the reading frame of 83% of Duchenne patients (Treat NMD, 2009). Professor Steve Wilton has gone on to show that it is possible to sequence every single exon skip required to treat 83% of patients with a relevant AO (Wilton et al., 2007). 75% of patients have a deletion that occurs in the hotspot region between exon 43 and 55. Skipping the top 10 exons - 51, 45, 53, 44, 46, 52, 50, 43, 6 & 7 - would restore dystrophic expression for >40% of all patients (Rus et al., 2009). Parents and patient organisations have so far backed the strategy for completing trials that will give compelling evidence for the use of one exon (exon 51). PTC Therapeutics has also announced recently that it is undertaking a Phase III study in a number of US and European centres to further trial its drug Ataluren. This drug targets a small sub-population of Duchenne patients that have single base pair gene variations in the gene. Even a single point variation can result in an out of frame mutation resulting in no dystrophin. It is proposed that Ataluren acts by skipping over this tiny mutation to produce functional dystrophin. Other exon skipping programmes are also underway using other chemistries, and while not yet at the human clinical trial stage, offer real promise for near-future application. These include conjugated AOâ&#x20AC;&#x2122;s developed at Oxford and Cambridge. The Oxford and Cambridge teams have won grants from Wellcome and MRC to continue the work first established with Action Duchenne funding. PNA chemistries are under

investigation and Action Duchenne has funded projects in the UK and China. Gene Replacement For those young people who are unable to benefit from antisense oligomers >20% skipDuchenne proposes an alternative strategy using gene replacement delivered via vectors. There is animal data to show that this method of delivery using AAV vectors could replace the faulty gene with a gene that could either promote exon skipping (AAV U7), replace the gene with a mini gene, or use a combination of vectors to restore the full gene at the membrane. Action Duchenne is already funding research in this promising area of gene delivery with Royal Holloway University of London. The skipDuchenne research project will build upon this and other research and aim to bring a genetic treatment to those patients unable to benefit directly from AO exon skipping in the next five years. Utrophin Upregulation Professor Kay Davies at University of Oxford has pioneered an approach to develop an effective therapy for DMD through increasing the amount of the dystrophin-related protein utrophin in muscle. Utrophin is a protein that is present in muscle cells at the junction where the nerve meets the muscle cell.

They have already shown that utrophin can functionally replace dystrophin in the mouse and dog models of the disease. Upregulation of utrophin has the advantage of being applicable to all patients as it is not mutation-dependent. Furthermore, a small-molecule, orally

administered drug can be more easily delivered to all affected muscle types including heart and diaphragm, and would not require the use of an immunosuppressant. N=1 Clinical Trials The current clinical trial programme for assessing the efficacy and safety of exon skipping antisense oligonucleotides follows the well established pathway of development. This involves the familiar phases of trial development through pre clinical, Phases I, II, III and IV, to drug marketisation. However, Action Duchenne, along with many others involved with the development of genetic medicines, question whether this framework is now fit for purpose for a rare disease like Duchenne. Duchenne patients do not present as a homogenous group. Every child will have as a starting point a different dystrophin gene variation that has an impact on the clinical symptoms of the disease. Every Duchenne patient will have polymorphisms in all manner of other genes that may impact on the progression of the condition. Crucially, every child will be exposed to a complex array of environmental, class and cultural factors including diet, housing, education, parental expectations and known medical interventions and support. Therefore, in clinical trial experiments, it is hugely difficult to achieve any reliable controls within the Duchenne population. It is highly likely that in a clinical trial, patients, for all the above reasons, will respond differently to any one drug. The current drive towards personalised medicines by drug companies is testament to the fact they have been unable for decades to find drugs that will treat a patient population in a uniform way. Consistently they have had to stop expensive trials because they have not been able to present uniform and consistent clinical data for a drug. The slow and expensive clinical trial programmes for drug development for Duchenne are not just frustrating families, they are not giving us the data we need. There is also an elephant in the room. Many families who see little hope in this INTERNATIONAL PHARMACEUTICAL INDUSTRY 79

Clinical & Medical Research process for their children are turning in increasing numbers to the internet for known drugs, supplements or devices that might help, and they would certainly buy AOs if they could. These are not crazy folk, but many who have seriously researched the subject and have the cash to buy drugs and devices online. So far the regulatory authorities have been unable to stop this trade, and indeed it is growing faster. Clinicians, researchers and GPs have turned a blind eye or simply do not know that their patients are taking online-sourced supplements or drugs. Apart from the obvious dangers of such experiments with drugs of unknown quality and guesstimate doses, it means that it is making it even more difficult to test a drug on a group of Duchenne patients. If patients taking these drugs or supplements were excluded from trials then we would certainly not have enough patients, as they are already thin on the ground. Combine this with the knowledge that some Duchenne patients are taking bisphosphonates, betablockers, ace inhibitors and steroids by agreed prescription, and the picture becomes even more complex.

The embryos of such databases do exist in the UK. The DMD Registry and North Star database have been great examples of beginning to collect this information for individual patients. Frustratingly, in the UK we do not have a database in the NHS that could do this for every intervention for every patient, despite massive expenditure on NHS IT infrastructure. However, this is an idea supported by clinicians such as Sir Gordon Duff in his Lancet article. Clinical experiments therefore become integral to a patient’s treatment regime. Also, designing trials that start at Phase III/IV or IV in this way also offers the potential for biotech companies to gain conditional market approval for a medicine while under extensive trial. This could be a real incentive for smaller biotech companies to recoup R&D revenue and support further development. Such trials should be driven by risk management, not a slavish adherence to existing phased protocols. Clinical trials could be seen as part of the clinical management process of every Duchenne patient, and this would be a marked improvement over the current system.

How can the Pharmaceutical Industry Help? We would suggest that rather than rejecting N=1 experiments, the answer lies in using as a starting point existing N=1 knowledge and patient data of every intervention (legal or not) and clinical assessment for the life of a single patient. New medicines can then be tested for safety and long-term efficacy for an individual patient, irrespective of their genome or current treatment regime. The research team’s job would then be to data mine all patients taking that new drug over an extended period, and begin to tease out patterns of medical interventions or genomic suitability that would seem to be working most effectively across the whole Duchenne community. We need to radically rethink clinical trial protocols for Duchenne, and we need a database that holds the medical life history of a patient.

skipDuchenne – Aims and Objectives Aims 1) T  o provide £5m of funding over the next five years to support the skipDuchenne research programme, that will aim to ensure that all patients living with Duchenne have access to a first genetic treatment in the next three to five years 2) T  o ensure that all those patients in the UK that could benefit from exon skipping using antisense oligomers (AO’s) - 83%, have access to AO drugs within the next –three to five years 3) T  o support the development of an alternative gene replacement technology using vectors that can deliver a gene therapy for those unable to benefit from AO exon skipping


Objectives 1) To work with biotech and iDESC partners to ensure that by 2014 all potential human exon skipping sequences have been optimised 2) To fund further research and work with biotech partners to collect experimental data to show the potential for using two AO drugs to promote double skipping (eg using 51 and 45) 3) To bring together biotech companies, research groups, UK government and the UK and European regulatory authorities to agree funding and a strategy to roll out all exon skipping drugs that could benefit 83% of Duchenne patients directly following the publication of compelling data for the market approval of exon 51 by 2014. 4) To support efforts for regulatory approval for clinical trials using AAV vector technology to deliver gene replacements for patients in the UK. 5) To fund further pre-clinical research and trials for the development of AAV vector gene delivery for patients over the next five years 6) To continue to support alternative ways of delivering exon skipping drugs and enhancing delivery

Nick Catlin is founder and Head of Research of charity Action Duchenne. The charity was set up by Nick and his partner Janet Hoskin and other Duchenne families in 2001 to support and promote innovative research into a cure and effective medicines for Duchenne Muscular Dystrophy. Nick and Janet have a son Saul, aged 11, who has Duchenne. Email:

Summer 2012 Volume 4 Issue 3


Reverse Osmosis for Laboratory Use Poor specification and maintenance can significantly reduce the quality of output from reverse osmosis (RO) water purification systems. In contrast, a well-specified, well-managed RO water purification system from a trusted and experienced supplier can deliver to the laboratory an efficient, economical and reliable supply to an exceptionally high level of purity. Water is typically the most commonly used element in the laboratory, and in many applications the quality of supply can be critical. As technology has advanced and instrumentation has become ever more sensitive and complex, water purification systems have likewise advanced to reduce contaminates present in the laboratory water to lower levels than ever before. In recent times, reverse osmosis has become established as an excellent option for water purification in the laboratory. Choosing, operating and maintaining an RO system to provide a high purity of supply should be straightforward, but systems are often incorrectly specified for the application or subject to improper use. However, a well-specified water purification system from a trusted and experienced supplier will provide an exceptionally high level of purity and, coupled with ongoing technical support and advice, will ensure that your laboratory receives an efficient, economical and reliable supply of purified water. To get the best results, laboratory technicians need to understand how to choose between standalone and centralised systems and appreciate the key issues that must be considered when selecting an appropriate system. Using that knowledge, technicians can take full advantage of the latest technology, which can significantly help to improve water quality and laboratory productivity, provided that basic process and operating guidelines are followed. The first stage in the specification process is to ensure that everyone 82 INTERNATIONAL PHARMACEUTICAL INDUSTRY

involved fully understands the grades of water purity that have been standardised, the available methods of delivering them and the requirements of the laboratory itself. Reverse osmosis is capable of producing purified water, in large volumes, from a supply fed under pressure into a module containing a semi-permeable membrane, which removes up to 98% of inorganic ions, plus virtually all colloids, microorganisms, endotoxins and organic macromolecules. Depending on the quality of the feedwater, RO systems are able to provide typically Grade 3 purity purified water. There are three grades of water purity, expressed in terms of conductivity, defined in the current standard, BS EN ISO 3696:1995 ‘Water for analytical laboratory use’. Grade 1 represents the highest level of purity, 0.01mS/m (0.1µs/cm), with Grade 2 being

0.1mS/cm (1µs/cm) and Grade 3 at a level of 0.5mS/m (5µs/cm). However, it is also possible to achieve higher levels of purity by employing a combination of reverse osmosis and deionisation. In addition, pre-treatment of the feed stream will protect and extend the life of reverse osmosis membranes, especially in areas where the feedwater contains high levels of organic contamination, hardness and free chlorine. As an alternative to resin-based deionisation systems, larger integrated laboratory systems can incorporate an electro-deionisation system (EDi), for secondary purification when fed with permeate from the RO system, producing water with a quality of greater than 10 MΩ.cm. Essentially EDi is a purification technology that uses a combination of ion-exchange membranes/resin and electricity to deionise water. Summer 2012 Volume 4 Issue 3


LABS & LOGISTICS For stand-alone systems, there are further measures that can be taken to enhance the quality of supply. For example, where Grade 1 water with enhanced microbial quality is required, the RO/deionised purified water is further processed using UV irradiation at 254nm and sub-micron filtration between 0.2 and 0.05 microns to remove bacteria and fine particulate matter. However, although the selection of the deionisation cartridges is relatively straightforward, the specification of RO systems that can achieve these enhanced levels of performance can be complex if optimum levels of performance, energy efficiency and operating costs are to be achieved. Even though there is potential to raise the quality of purified water supplies delivered by stand-alone units, there are situations where it is more efficient and economical to install a centralised system. For example, as most purification systems will have to comply with BS EN ISO 3696 and provide water on demand wherever they are needed throughout the lab, the nature of the work may dictate that one or more self-contained units need to be positioned at different locations. Space being a valuable commodity in most laboratories, the chosen water purification solution needs to be as unobtrusive as possible whilst still delivering the required quantity and quality of water. In these situations, a centralised system feeding a ringmain may be more appropriate. Once the benefits of stand-alone or centralised water purification systems have been considered, it is time to choose and specify a system that will achieve the right level of water purity and the most effective balance between cost and efficiency. A common cause of inefficiency in water purification systems is oversizing, so it is important to specify equipment that can deliver only the volume of purified water that you need. Oversized systems require extra space, greater expense and, because RO is potentially less efficient when the laboratory is only operational for short periods, can suffer degraded performance. Likewise, a realistic estimate must 84 INTERNATIONAL PHARMACEUTICAL INDUSTRY

be made of the number of take-off points that will be in use at any one time; if it is simply assumed at the installation stage that all points will be in use at once, the result can be a dramatically oversized and expensive system. To maximise efficiency, it is essential to consider whether a given water quality is required throughout the laboratory, or only at a limited number of work areas. Similarly, the volume of water needed should be analysed based on the patterns of daily use to highlight peaks and troughs in water requirements over extended periods. To achieve the best specification

it is advisable to work with a supplier who is willing to assist you on-site and help you to specify the best solution for your needs, and one that also has a capacity not only to supply but to plan, specify and install a system, rather than delegate to outside contractors. With the right help and advice, an efficient, economical and reliable supply of purified water is available for your laboratory that will remain costeffective throughout its lifespan. It is also important to consider the fact that all water purification systems require routine cleaning and maintenance to ensure consistent levels of performance and reliability. Summer 2012 Volume 4 Issue 3

LABS & LOGISTICS The key is to choose a unit that is quick and easy to maintain with easy-tochange consumable parts. Similarly, the cost of consumables should be taken into account, as systems that use high volumes of resins, chemicals and cleaning solutions can quickly become uneconomical. It should be remembered that, although an RO system may be able to deliver the total volume of water required over time, it may not be able to meet sudden surges in demand, so sufficient purified water storage facilities must also be considered. Regular quarterly cleaning of RO membranes is advised. This is relatively straightforward and is typically carried out using specialised cleaning solutions. Acid-based cleaners are used to remove scale, and alkaline-based solutions to remove organic matter; if required, special chemicals can also be used for disinfecting, but should only be added once all scale and organic matter has been removed. Solutions are simply circulated through the RO system and then flushed to drain. Regular maintenance will ultimately reduce operating costs by preventing unexpected downtime or component failures. Costs can be reduced still further by recovering the concentrate stream for use in low level duties such as plant wash-down, vehicle cleaning or toilet flushing. Whether you choose a standalone or a centralised system, there is one aspect of reverse osmosis water purification technology that is continually being enhanced to the benefit of all users and that is the continuing refinement of the semipermeable membrane elements. Advances in membrane technology have allowed pump speeds, and correspondingly pump pressures, to be significantly lowered, thus reducing energy levels. In centralised systems, it may be possible to make further gains in efficiency if pumps are linked to variable speed drives, enabling the speed of each pump to be matched exactly to the output demands of the process and water treatment system. Improvements in the efficiency of membrane elements include the development of extremely thin membrane materials, 86 INTERNATIONAL PHARMACEUTICAL INDUSTRY

some 120micron in thickness, that are based on polyamide thin film composites with non-woven polyester support webs. Elements are manufactured using advanced adhesive techniques, allowing both the spiral-wound mesh space between layers and the layer-to-layer bonding areas to be reduced. This is achieved without compromising the thickness of the feed spacer or affecting the mechanical or chemical properties of the membrane. As a result, the feed pressure, compared with a traditional high rejection RO element, has been significantly reduced, with lower fouling potential and less pressure-drop, while flow rates have increased dramatically. This also means that the operating life of many applications can be significantly extended, while the creation of a far higher active membrane surface area has given designers the option to reduce the total number of modules in larger reverse osmosis systems and reduce the need for repairs and maintenance. In summary, laboratories can enjoy high levels of water purity using today’s reverse osmosis systems, and further increase efficiency and purity over bottled distilled water or on-site distillation units by fitting disposable deionisation or ion exchange cartridges directly to a mains water supply. Stand-alone RO units can produce water of Grade 1 purity when permeate is circulated through a deioniser, while larger integrated systems can incorporate an electro-deionisation system (EDi) for secondary purification when fed with permeate from the RO system, producing water with a quality of 10 to 15MΩ.cm, depending on flow rates

and the quality of the feedwater. Perhaps the most important fact of all to bear in mind is that although there are many companies with the ability to supply, install and commission RO systems, there are far fewer with the resources and expertise to help end users maximise the return on their investment once the installation and commissioning engineers’ work is complete. To help you make the right decisions regarding specification and maintenance factors from the outset, choose a supplier who is willing to work with you on-site and help you to specify the best solution for your needs. A well-specified system, coupled with ongoing technical support from an experienced supplier, will ensure that your laboratory receives an efficient, economical and reliable supply of purified water over a long system lifespan.

Mark Bosley has worked for Purite for over 15 years, and is currently employed as their Business Support Divisional Manager, heading up the company’s Technical and Industrial/Laboratory Systems contracting. In total, Mark has over 30 years’ experience in the water treatment industry. This has included work in the development of both standalone laboratory water purification units and the design of specialised purified water systems for a variety of market sectors, including brewing and distilling, food processing, healthcare and pharmaceutical. Email

Summer 2012 Volume 4 Issue 3




Answering Pharma’s Need for Outsourcing – Process ‘Intensification’ Technology for the Future of Biopharmaceutical Production The chemical and pharmaceutical industry will face challenges in the future related to manufacturing costs and lowering environmental impact. Chemical and biopharmaceutical manufacturing need to be modernised, with lower raw material usage, higher yields with fewer resources, more focused manufacturing processes, and reduced manufacturing cost as final results. DSM Pharmaceutical Products, the outsourcing manufacturing partner within Royal DSM N.V. (#1 on the Dow Jones Sustainability Index 2011), a global sciencebased company active in health, nutrition and materials, is a custom manufacturing organisation offering proprietary (bio)manufacturing technologies, innovations in sustainability, and green chemistry, in R&D and commercial manufacturing services. Application of technologies brings down its manufacturing costs through process intensification to offer shorter and more efficient scaleup of (bio)pharmaceuticals, speeding up development, better managing manufacturing volumes and driving down total costs. In this context DSM challenges conventional process technologies with innovations in process intensification for next generation manufacturing solutions. These innovations range from boosting cell culture titers and utilising alternative starting materials in synthesis routes to increase drug substance yields and continuous flow API production. Biotechnology and (bio)chemistry are applied when working with microorganisms, enzymes, mammalian cells and organic starting materials to produce novel ingredients and to improve existing ingredients, (bio) pharmaceuticals, and chemicals. With the market facing downsizing in production volumes of certain APIs, smaller batches of high-powered speciality drugs and a need to reduce costs and environmental impact, DSM has invested in multiple synthesis, scale-up and production technologies continuously over time. Especially in the pharma 88 INTERNATIONAL PHARMACEUTICAL INDUSTRY

outsourcing market a global portfolio of resources is needed to continuously serve changing customer needs and bring real value as the pharma industry shifts business models. In today’s market where manufacturing efficiency has become a strategic driver for pharmaceutical companies, contract manufacturing organisations like DSM are responding to provide sustainable solutions with their manufacturing expertise, advanced technologies and regulatory support. Biomanufacturing (Mammalian and Micro-organism-based) The three major economic drivers for the development of protein therapeutics manufactured using mammalian cell culture are the cost of goods, time to market approval, and reduction of financial risks associated with capital investments for dedicated production facilities. In mammalian manufacturing, the trend for process intensification has been developing for many years. The productivity of cell lines has been improved by using better production vectors, targeted integration into the hot spots and better use of cell metabolism. In addition, modern media supports higher cell densities in standard fedbatches, increasing yield output. One way to address these drivers is to improve the efficiency and productivity of the manufacturing processes. This is relevant for new biological entities but even more relevant for biosimilars and biobetters manufacturing. DSM has developed a strong technology portfolio (Figure 1) that enables Figure 1. Flow chart: reduced number of unit operations, intensified processing, and increased output from decreased footprint are all advantages of DSM’s XD® and RHOBUST® technologies.

shorter processing times and a lower cost of goods to achieve this goal. In addition, the manufacturing facility footprint is significantly reduced with the use of these technologies, thereby reducing capital investment and enabling easier investment decisionmaking. Today 5g/l for antibody production is standard. However, DSM has taken yields to higher levels with a manufacturing process to achieve 5- to 15-fold increases. For example, XD® Technology leads to very high cell densities in a bioreactor, giving significant titer improvement compared to standard fed-batch processes. The XD® process technology combines the benefits of fed-batch and perfusion processes. This technology works in a continuous media feeding mode, with a filtration unit to retain both the cells and the recombinant protein in the bioreactor, while metabolic by-products are continuously washed out. It is demonstrated to be virtually cell line and product independent, as it has been successfully applied to multiple cell lines (CHO, hybridoma, myeloma, PER.C6® cell line) and to multiple product types (monoclonal antibodies, Fc-fusion proteins, recombinant proteins). The XD® process is robust and scalable, while still maintaining consistent product quality. The scaleup of XD ® cell culture is possible after minimal process development, which significantly reduces the timelines and costs of the development. RHOBUST® is a direct protein capture downstream technology which combines clarification and product capture into one single step. Centrifugation, depth-filtration and packed bed chromatography can be replaced by one unit operation, resulting in less preparation and process time. RHOBUST ® uses cross-linked agarose beads with tungsten carbide to increase the particle density. Cell removal through centrifugation and/or filtration is made obsolete due to the fact that the crude harvest can be loaded directly onto the RHOBUST® column. Cells flow Summer 2012 Volume 4 Issue 3

MANUFACTURING through the column, while the product is being captured on the beads. As an additional effort to achieve further streamlined purification and polishing of proteins as a single unit operation, DSM has developed the Kremer Method™. The Kremer MethodTM is an inline flow-through protein polishing step. This seriallylinked flow-through purification for IgG combines 2 chromatography steps in one unit operation (see Figure 2).

is on the market. Using large vessels to produce pharmaceuticals has a number of disadvantages. Most importantly, large-scale chemical reactions in these large vessels deliver large quantities of unwanted – sometimes dangerous – byproducts. These by-products have to be disposed of, resulting in a negative environmental impact. The traditional production method – which also requires heavy use of energy

Figure 2. Kremer Method™ - Serially-linked flow-through purification for IgGs after product capture.

For example good removal of HCPs (12,000 ng/mg to less than 31 ng/ mg using a Mustang Q membrane absorber) and aggregates (3.6% to 0% using Poros HS CEX column) were observed using a ProteinA purified IgG from mammalian source as start material. Advanced technologies in microbial fermentation-based processes are continuously developed and improved for in-house production processes at DSM. Innovations range from production strain construction and improvement, to fermentation process concepts, and product isolation technologies. These innovations fit classical fermentation products as well as modern biotechnologybased products such as recombinant proteins, advanced scale compatible expression platforms for protein production, various modes of fermentation process execution, and select high-performance product isolation and purification operations. Synthesis Optimisation and Micro-reactors Adopting a micro-reactor production platform requires that an outsource partner can demonstrate the knowledge and skill in such manufacturing to take the process from the small scale to the scale required when a customer’s product 90 INTERNATIONAL PHARMACEUTICAL INDUSTRY

resources and can generate a lot of heat that needs to be cooled – is not very efficient; more solvents, catalysts and other starting materials are needed than is desirable. A micro-reactor is a continuous tube reactor with a small channel diameter. To reach the desired throughput of such a reactor, it contains thousands of small channels in parallel, each of them surrounded by coolant. In this way, the average micro-reactor can be as small as a small cupboard and can handle 1000-2000 kilograms of product per hour, with yields more than 20% higher than in the traditional large vessels, with a much lower CO2 footprint and at lower costs. Measured per cubic meter (kg/m3h) the productivity increases by a factor of 103-104. DSM has been working with microreactors for more than a decade, and in early 2009 completed full-scale commercial production of an active pharmaceutical ingredient. We practice 98% of prevailing chemistries and create many active ingredients and intermediates. In our unique convergence of biocatalysis and chemocatalysis, or “innovative synthesis”, we are finding faster routes to new drug compounds with more renewable starter materials. DSM route scouting services meet outsource needs for the development

of robust low-cost manufacturing routes. Route scouting capabilities have proven to lead to significant cost savings and a better environmental footprint by the reduction of synthesis steps or redesign of synthesis routes, where true innovation is achieved by integration of different cutting-edge technologies developed by DSM and partners. DSM Pharmaceutical Products business group serves the global pharmaceutical and biopharmaceutical markets, offering manufacturing services, R&D/ formulation and technologies for biologics, both cell culture-based and microbial; active pharmaceutical ingredients and intermediates; as well as finished dosage manufacturing. The business group works closely with all DSM pharma interests including DSM Sinochem Pharmaceuticals, Biomedical, and Nutritional Products. Dr. Fritjof Linz Senior Director New Business Development & Marketing, DSM Biologics Dr Linz started his business career in the downstream processing industry holding different roles in sales, product management and development at Sartorius. Thereafter, he became Head of Marketing & Sales for a microbial scale-up CMO – Devoferm which is a member of the Aventis Research & Development group. From here, Dr Linz moved to a facility & site management organization called InfraServ, which serves the Pharma & Biotech industry. Dr Linz joined DSM eight years ago having responsibilities for large Pharma companies’ chemical and microbial CMO business, serving countries like Germany, Japan and others. For the last four years he has been a member of the DSM Biologics sales team with responsibilities for custom manufacturing and technology licensing around the world – mainly Europe. In addition he is the Head of Marketing for DSM Biologics. Dr Linz has a PhD. in Chemistry at the University in Hannover with Biotechnology as the major subject. Email:

Summer 2012 Volume 4 Issue 3


The Healthy Printer: the increasingly powerful levels of functionality and added resilience offered by the latest label, barcode, portable and point-ofsale printers can offer long, troublefree life to serve busy pharmacies.

The printing of prescription, identification and barcode labels takes place every day in every pharmacy throughout the country. However, busy pharmacists probably never give a second thought to the printer they use, until it goes wrong, or unless it regularly causes problems such as jammed media and software incompatibility. Problems occur for a number of reasons, including incorrect hardware or label specification, so it pays to understand and appreciate current developments in printer technology and thus consider which type of printer is best for a given application. Armed with this knowledge, pharmacists can then select machines based on the right criteria to ensure long life and trouble-free operation. Today, barcode and label printers come in all shapes and sizes, enabling businesses large or small to benefit from the ability to print fast, highquality labels. Indeed, these machines are now such an integral part of many businesses that employees and operators often take them for granted. This is a testament to the success and functionality of barcode and label printers, but the downside of this success is that the units are not always treated with enough care or specified to provide the most suitable service for the application concerned. With this in mind, it is important to consider the enhanced levels of functionality and defence against failure offered by the latest units, particularly given the 92 INTERNATIONAL PHARMACEUTICAL INDUSTRY

cost of faulty or inefficient printers to a business that uses them day in day out. In the pharmaceutical sector it is also crucial to be aware of current regulations. From the Cosmetic Products (Safety) Regulations to guidelines released by the Medicines and Healthcare products Regulatory Agency (MHRA) and the Food and Drug Administration (FDA), we have witnessed the introduction of a growing volume of complex legislation that impacts on the packaging and process operations of cosmetic, healthcare and pharmaceutical products. As a result, labels have become an increasingly critical element in the process, acting as a mechanism for delivering a wide range of variable data for efficient product identification and of adding key information for end users. As the majority of todayâ&#x20AC;&#x2122;s cosmetics contain complex mixtures of industrially produced, synthetic chemicals and naturally occurring compounds, it is vital that labels on all such products contain important information so that consumers with allergies, for example, can identify which products to avoid. Equally, pharmaceutical products must carry labels that state expiry dates, precautions or warnings and dosage instructions to ensure maximum consumer protection. Happily, todayâ&#x20AC;&#x2122;s barcode and labelling machinery is increasingly more resilient and easier to use, helping users to address many of the issues

described above by offering more reliable production and higher levels of performance to meet regulations. Many printers are designed and built specifically to meet the demands of the sector, with strong in-built safety and reliability features that require minimal maintenance. This has resulted in a reduction in POS delays and machinery repair costs, both of which can dent profits. Designers and engineers have carefully considered the kind of problems that develop when a printer is dropped or becomes dirty, such as faulty connections and failures in printer communications. They have also considered the problems that are caused when untrained operators use incorrect tools or adopt a heavyhanded approach when dealing with simple paper jams, especially when under pressure or at busy times. These considerations have provided some strong solutions. Taking a broader view of the technology as a whole, one recent innovation that has proved to be most powerful is the GS1 DataBar code, which is fast becoming the global standard in the sector thanks to its impressive functionality. For example, these codes can hold considerably more information than standard barcodes, while also supporting Global Trade Identification Numbers (GTIN) for variable data such as batch, expiration date, serial number, price, monetary value, size and weight. Summer 2012 Volume 4 Issue 3

MANUFACTURING The introduction of these latest innovations looks set to have a big impact on the efficiency, traceability and safety in the pharmaceutical, healthcare and cosmetics sectors, however, it is important to recognise that these improved levels of performance can only be achieved when using the right printer technology. Although there is a wide range of printers already available to print the latest barcodes, not all have proved capable of operating reliably for extended periods of time in busy environments. However, market leaders have succeeded in providing features that can satisfy this need. For example, the latest portable models use a fail-safe, easyto-use, drop-in paper loading system that dispenses with the usual fiddling involved with reloading; the media is simply dropped into the paper holder and the lid closed on the leading edge of the paper in a matter of seconds. With regard to printing GS1 DataBar codes, the latest printer technology is

able to accommodate different types and sizes of media. As well as being suitable for extremely small labels under 2.5cm in width, the new barcodes can be printed on specialised media, including antimicrobial polyester labels, plastic substrates and metal tags, as well as on conventional paper, making them ideal for small or irregularly shaped products. The compact GS1 DataBar barcode is deceptively simple in appearance and reveals little of the additional data-carrying capability it offers. Nevertheless, DataBar not only encodes the pack price and product type but also the weight of items, and it is this information that, when used in stock management systems, can help indicate when replenishment orders are needed to ensure correct stocks levels are maintained. Another feature of GS1 DataBar is the encoding of a sell-by-date limit, so that perishables are sold within a specific time period and are actually prevented from being

sold when that period has lapsed. GS1 DataBar brings benefits to the cashier, too. The high-density encryption techniques used in DataBar enable the production of a compact barcode that ensures ease of scanning with packaging when placed in front of flatbed scanners. Item recognition is possible when orientated in any direction, which streamlines counter service in pharmacies to the benefit of both the chemist and the customer. These barcodes are also compliant with the latest barcode and label printers and can be printed with both existing EAN and GS1 DataBar formats. Indeed, the modern barcode and label printer includes a host of features to complement the GS1 DataBar, offering ease of integration with a wide range of existing applications or networks, as well as a cross-emulation function for different programmes. The machines are now more robust, faster, and offer the versatility of printing in both direct thermal and thermal transfer



modes that can print the compact GS1 barcodes labels up to 300 dpi. To eliminate the risk of error, some of the latest printers also incorporate inline validation systems that automatically check and validate all printed labels for accuracy and possible degradation. Therefore, quality control and compliance with standards can be monitored quickly and reliably as each label is produced. If print quality drops, for example, as a result of contamination entering the printerâ&#x20AC;&#x2122;s casing and obstructing the print-head, an alarm is activated to alert staff, enabling them to remedy the problem as soon as possible. Likewise, if the print-head needs replacing, staff are made aware sooner rather than later, allowing the printer to remain operational. The issues for larger desktop printers can, of course, differ from those faced by small POS units, but the advances made regarding these devices have also made a positive impact on efficiency. Common, simple errors that, nevertheless, consume valuable time can be addressed more swiftly than before. Convenient, tool94 INTERNATIONAL PHARMACEUTICAL INDUSTRY

free maintenance systems offered by modern units have been a great benefit to the smooth running of operations; for example, self-clearing cutters keep POS systems running without delay in the case of a cutter jam. Similarly, new options to painlessly replace auto-cutter and printer head components via vertical flip-top opening mechanisms, or load ribbons safe in the knowledge that automatic adjustment systems will ensure the correct tensioning, have been warmly welcomed. As a further insurance against downtime and repair costs, the leading printer manufacturers offer extended warranties of three or even five years, and have comprehensive dealer networks to provide technical support, ensuring that retail businesses run smoothly and efficiently. Such issues as those described above should dissuade readers from buying cheap barcode and label printers, a decision that can swiftly prove to be a false economy. This may not be a new argument but it remains an important one, especially when traders in all industries are fighting hard to manage costs and maximise

profitability in a difficult economic climate. The difference between lowcost printers and those from leading brands is small, but that small saving can cost many hundreds of pounds in lost revenue and damage to brand integrity when printers fail. It is also worth bearing in mind that if barcode labels are being produced then it is common for rigorous standards to be applied regarding factors such as the thickness of bars, bar spacing, the ratio of narrow to wide elements and the levels of contrast between dark and light elements. If these standards are not met, for example due to a printing problem, then there is a significant risk of rejected labels or packaged items. Companies invest considerable time and resources in developing machines that will withstand the toughest conditions, ensuring that printers are not built to fail. Tests range from dropping printers on the floor, through to extended lifetime and environmental testing of print mechanisms and electronics. Some companies and their distributors and resellers also understand and advise Summer 2012 Volume 4 Issue 3

MANUFACTURING on the specification and management of printers because, in practice, most printer failures generally arise from incorrect specification, improper use and poor standards of maintenance, all of which can result in a reduction in print quality, problems with print-heads and cutters, and overheating of control boards and power supplies. Correct specification depends on a number of factors. Although some may be specific to a particular application, they will generally include: the volume of labels required and the frequency of operation; the operating environment, in terms of temperature, humidity and levels of dust and other contamination; the type of labels being produced and the label stock; connectivity to other equipment; and lifetime cost of ownership, based on energy consumption, media costs and any service requirements. Perhaps the most common cause of printer damage is exacted by users who, under pressure of a heavy workload, adopt a heavyhanded approach to releasing paper jams, as described above. Internal mechanisms that have been designed to produce fine quality printing are necessarily sensitive and not equipped to withstand brute force. Of course, although too much handling can cause complications, neglect can be just as bad. If maintenance and servicing is not performed at the prescribed intervals, then weak printing, wrinkled ribbons and, inevitably, printer failure will soon follow. Without appropriate specification, installation, handling and maintenance, even the finest machinery can fail early. However, with the correct printer specification and the right attention paid to operational and maintenance requirements, today’s printers can prevent those unwanted interruptions in your service that disappoint customers, protecting profits by offering long, reliable service. 96 INTERNATIONAL PHARMACEUTICAL INDUSTRY

The options available to operators of barcode and labelling machinery are many, and a little time spent selecting the most appropriate printer specification can save a lot of time and money. Modern barcode and labelling machinery can cut maintenance costs and deliver such a powerful performance that the investment is more than returned by protected revenue and the smooth running of the business. With the pressures of new labelling regulations, and competitive global market pressures, the use of functionality of labelling equipment is progressively evolving. The recent developments in labelling technology are enabling pharmaceutical, healthcare and cosmetic packaging companies to improve both productivity and profits, as well as enabling them to be fully compliant with the latest regulations to ensure ultimate protection for consumers.

Jörk Schüßler Educated in Germany, Jörk Schüßler is European Marketing Manager for Citizen Systems Europe, responsible for their activities across Europe, the Middle East and Africa, and has 20 years of experience within a sales environment including sales and marketing in hardware, software, e-learning and TV. In this time, Jörk has held several senior management roles, including ten years at Citizen, and is now set to develop Citizen’s support infrastructure for its range of distributors, value-added resellers and end users in many different sectors, and is part of a wider longterm company strategy. Email:joerk.schuessler@

Summer 2012 Volume 4 Issue 3


A Fluid Process A wave of new and modified flow technologies has helped to create effective solutions for manufacturers of medical devices. However, the maximum benefit of these advances can only be gained by working alongside an expert provider of fluidic technology from the early design stages, says Sherea Lizaso, Medical Marketing Manager for Gems Sensors & Controls. The control of fluid flow â&#x20AC;&#x201C; both gas and liquid â&#x20AC;&#x201C; forms an essential part of many different types of medical equipment, from boiler control on steam sterilisers and reagent dispensing in in vitro diagnostic equipment, to bulk delivery of gases or precise gas delivery in products such as ventilators. Increasingly, the drive is towards reduction in the size and weight of equipment, while at the same time increasing performance and reliability. For manufacturers, however, these demands create a dichotomy; for example, reducing size can have a significant effect on flow rate and dynamics, while component integration can lead to an increase in complexity and potential failure rates. However, a wave of new and modified flow technologies, in conjunction with a fresh approach to equipment design, has helped to create effective solutions. Specific examples of product development illustrate this; for example a recent redesign of flow valves and manifolds on a transport ventilator has helped to reduce space requirements by 40%. A major challenge for the providers of flow solutions for medical devices is to supply laboratories, hospitals and physicians with these increasingly complex yet compact systems while managing cost both internally and for customers. This has driven manufacturers to develop new design and production methods that deliver on all fronts to the benefit of both manufacturer and end user. For example, a smaller unit footprint represents a market opportunity for a manufacturer as it will be a 98 INTERNATIONAL PHARMACEUTICAL INDUSTRY

popular choice for many commercial laboratories; these businesses are often measured by revenue per square foot, and technicians will therefore be inclined to choose more compact instruments when specifying equipment. Small, portable devices will also be preferred purchases for emergency medical services where equipment size is a key factor. Size can be effectively reduced by consolidating components that are typically separate constituent parts into a single functional unit. This was recently proved in collaboration with a producer of respiratory equipment that wanted to offer a compact transport ventilator. When working on the gas mixing system for the ventilator, which controls the concentration of oxygen and room air that is delivered to the patient, the challenge was to reduce the physical size and power consumption of the blending system while still maintaining requisite flow parameters to comfortably manage oxygen delivery for patients ranging from neonates to adults. The necessary size reduction was achieved by challenging the design of the existing gas blender. For instance, the existing system relied on needle valves that had to be manually adjusted to set the gas flow to a precise range; in contrast, the redesign of the system employs precision orifices that are pressed into the manifold and are controlled externally. The need to access valves was thus eliminated and the amount of space required to house the manifold was significantly reduced. This solution has the added benefits of decreasing the total number of parts and eliminating labour-intensive calibration. To reduce the size further, further functions were taken that had been located externally and relocated inside the manifold. Specifically, the nebuliser block (used to moisturise the gas mixture) and temperature sensor were placed within the body of the manifold block. With access constraints eliminated, the overall footprint of the device was reduced by 40%.

The success of this project was the result of carefully applied expertise and, though the solution may appear to be simple, this impression is deceptive. Reducing the physical size of equipment while still maintaining requisite performance is a difficult task and can introduce complications of its own, which is why it is so important for specialist technicians to collaborate in the early design stages. For example, where some equipment manufacturers have opted to produce smaller renal dialysis units without entering into an appropriate period of consultation with expert technologists, the benefit of gaining a small degree of added floor space around the unit has been offset by the complications this has introduced during maintenance. The optimisation of medical instruments and equipment always needs to strike the right balance between issues such as size and ease of maintenance, and it is the capability of component and equipment manufacturers to meet challenges such as this that ultimately wins the confidence of healthcare technicians and provides secure, reliable service for patients. A business benefit of working with a specialist provider of fluidic solutions is that it frees up valuable time for staff who can then focus solely on maximising the performance of proprietary components or systems, rather than diluting their priorities with non-core technology that is not their forte. In small companies, where cash flow is especially critical, money can be saved by working with an external fluidics partner because although this will add a cost to the project it is nevertheless more cost-effective than employing permanent staff. A further benefit is that the cutting-edge expertise of a specialist partner is more likely to deliver an innovative solution to product development challenges and to do so with greater speed. Faster development will, in turn, mean a more efficient, focussed production process that achieves the best results and satisfies the specific needs of the marketplace. Design of the fluidics Summer 2012 Volume 4 Issue 3



MANUFACTURING component can proceed in parallel with other system functions, enabling designers and engineers to focus on the development of their specialist area while sharing feedback with other teams. The process can result in the fluidics specialist team delivering a single, fully-constructed subassembly that is tested and ready, and after the product has been built the fluidics team can provide an expert source of support if performance issues arise. Clearly, the key to a successful design and engineering project in this field is to involve a fluidics expert early in the development process. This not only speeds development but prevents wastage of time and resources at the end of a less collaborative design process when awkward and expensive changes can be required. By understanding the limitations and potential of components at the earliest stages, the most effective design can be achieved and any compulsory regulatory review, testing or validation that may necessitate modification can be considered and prepared for. Without an early consultation, project costs can significantly escalate; it is not uncommon, for example, that an OEM will approach a contract fluidics firm with a predefined specification and find that incompatibilities between the fluidics requirements and the overall product specification require an expensive and time-consuming exploratory pre-development phase. In summary, involving a fluidics design partner early allows design requirements to be discussed as the piece of equipment is being created, enabling many potential problems to be seen and sidestepped early on in the design process. This of course requires the free sharing of information, and naturally some parties are reluctant to provide such information to an external source, even in the course of progress. The best way to minimise concern here is to enlist the services of a partner that is equally transparent and regularly engages in consultation and knowledge-sharing with manufacturers. When all parties do share information and work closely together, the results are typically impressive. The technological expertise offered by the designers and 100 INTERNATIONAL PHARMACEUTICAL INDUSTRY

developers of todayâ&#x20AC;&#x2122;s sensors offers great advantages to OEMs in the construction of custom, engineered fluid systems and, while personal attention and technical support is available to produce bespoke systems to customersâ&#x20AC;&#x2122; individual requirements, it is also true to say that, in many cases, sensor manufacturers have now considered so many problems that they may already have a product solution available off the shelf, saving further time and money. For example, the performance of dialysis machines, which provide a critical service to patients who have lost kidney function by purifying the blood and preventing overhydration, has been streamlined via the use of advanced flow sensors. Owing to the extremely high temperatures required during sterilisation cycles, the slow reaction times of some sensors within these machines has triggered equipment shutdown, an often unnecessary safety measure that has brought inconvenience, delay and disappointment to waiting patients. This problem has now been addressed by the designers and engineers of flow sensors, resulting in the manufacture of ultra-compact flow switches that are not adversely affected by high temperatures. These sensors are equipped with a magnetic piston that is displaced by liquid flow to magnetically actuate a hermeticallysealed reed switch isolated within the unit. As we have seen, fluids play a role in many medical devices. From

precision instruments such as in vitro diagnostic systems, medical lasers, drug-infusion pumps or ventilators, to equipment such as steam sterilisers or systems for bulk delivery of gases that require large-scale fluid transfer, fluid management is critical wherever a liquid or gas needs to be measured, monitored or controlled. Among these devices there is a wide range of fluidmanagement needs, each with its own set of functional requirements. The concerted, consistent, dedicated effort of market-leading sensor designers to continually consult with clients to enhance and improve processes is bringing clear and measurable results to medical science, results that reduce patient discomfort and stress and enable more efficient and successful healthcare. Sherea Lisazo is Global Medical Marketing Manager of Gems Medical Sciences. Before joining Gems, Sherea occupied the position of Global Business Manager at TEMPIL, having progressed through the ranks of the company from her initial role of Marketing and Sales Development Specialist. Her business career has been conduced in tandem with a series of successful academic achievements, including a qualification as Executive Master of Business Administration in 2009 and a Masters degree in Science, Professional and Technical Communication during 2006. Email:

Summer 2012 Volume 4 Issue 3




Simplifying the Lives of Patients

Drug delivery devices currently feature among the major pharmaceutical trends. Their optimisation is largely influenced by the pharmaceutical industry, physicians and patients. Present developments of new devices are increasingly focused on patients’ needs. They give top priority to simplified handling, reduced pain, continuous documentation and facilitated adherence to medication intervals. A precise definition of drug delivery devices (DDD) is almost impossible to find. In general, DDDs support the transportation of active ingredients to the part of the body where they are needed and therefore can be considered as an application facility. Whether they are defined as pharmaceuticals or medical products depends on their purpose. For example, a powder inhaler qualifies as a pharmaceutical, while a catheter dissected with heparin qualifies as a medical product.

Pen Systems: Close to Perfection Due to a significant refinement of most DDDs in recent years, for some appliances currently very little potential for improvement is left. The introduction of pen systems in 1985 considerably enhanced patients’ quality of life. Pens are loaded using cartridges and usually serve as multiple application systems. Several functionalities such as level indication and haptic pen systems have already been optimised. Moreover, today’s pen systems feature electronic components like memory control, which is mainly an application for children. In future, further refinement will focus on easier and safer handling, such as, for instance, improved needle protection to help avoid injuries and unintended application. The optimisation of other parenteral systems also focuses on the patients’ compliance, reflecting the awareness of fears such as needle phobia. Autoinjectors are usually loaded using prefilled syringes and feature a needle cover keeping the needle hidden as long as possible. This often helps

users to overcome fears associated with self-injection. Diseases such as multiple sclerosis, osteoporosis, hepatitis, rheumatism and anemia are typical fields for the application of autoinjectors. Treatment of these requires a singular dose or regular administration during longer intervals and is profitable from the industry’s point of view. Therefore, intensive development of auto-injectors is not only justified to improve patients’ quality of life but also from an economic perspective. Further advancements mainly refer to production conditions. With optimised materials and designs, production costs will decrease. As a result, devices are no longer too expensive to be applied as reusable items. Some experts believe needle-free injection technology will soon prove to be successful. One of the reasons is the vaccine campaign launched by the World Health Organization (WHO). However, the pharmaceutical market remains cautious about these systems because they are believed to cause similar levels of pain as a puncture does. Transdermal Devices: More Convenience Transdermal delivery devices are patch systems containing active agents, for instance within matrices, which are diffused through the skin. Recent developments have led to optimised patch sizes and adherence features of the patches for better skin tolerance. A general trend in the western world goes towards the usage of and affinity for electronic devices, influencing also pharmaceutical applications. Exciting new developments comprise a combination of patch and pump, so-called “patch pumps”. This can be illustrated with the example of the Pancreum BetaWedge insulin pump. The small device with a bluetoothenabled personal diabetes analyser (PDA) adheres to the skin and


Summer 2012 Volume 4 Issue 3



PACKAGING delivers insulin for as long as three days. Electronic devices, such as technologically suitable MP3 players and memory cards, enable easy and convenient measurement of the blood sugar level. With “MyDose,” Roche is currently developing a single-use infusion device for the subcutaneous delivery of a new drug formulation containing monoclonal antibodies such as Herceptin and MabThera. The subcutaneous delivery is based on the patients’ individual bodyweight or surface. It is easier to administer, and application times are significantly reduced. The main functional component of the device is the “Vartridge,” a hybrid container which combines the features of glass vials and cartridges and is controlled via the electronics in the pump. As opposed to intravenous applications, patients no longer need to be treated in hospitals and are more independent in their everyday routine. Pulmonary Systems: Focus on Asthma and COPD Inhalable insulin seemed to hold a lot of promise. But the large size of the inhaler, increased treatment costs and the requirement for much higher insulin doses led to its withdrawal. The market for pulmonary systems has noticeably slowed down in developed countries, while it is still growing in the emerging markets. Yet pharmaceutical engineers continue with the advancement of these systems mainly with respect to asthma and COPD (chronic obstructive pulmonary disease). There are three different types of pulmonary systems. MDI (metered dose inhaler) is the most common technology in the market at low cost level, especially for emergency products. The aerosolbased inhaler delivers the required amount of gaseous medicine to the lungs. New products however are rarely developed in the form of MDIs for environmental and compliance reasons, such as patients’ coordination problems between breathing and actuation. The technology of the future – if no oral medication is available – is the DPI (dry powder inhaler) with its simple application and small size. Medication is delivered to the lungs in form of dry powder which patients inhale without any solvent. The third 104 INTERNATIONAL PHARMACEUTICAL INDUSTRY

pulmonary system is the nebuliser, which administers the inhalation liquid in the form of fine drops. Patients inhale the medication as a mist generated by different technologies such as ultrasound, piezo crystals or vibrating mesh. DDD – Key to Success of a New Drug Many established medical devices, and especially applications for children, have been significantly optimised over the past years. From a current perspective, some of them are approaching their improvement limits, while others still have much potential for further refinement. In general, the equipment tends to be smaller, and handling is safer, easier and more convenient. At the same time, costs are decreasing for those devices that hardly require any further improvement. In many fields, the market will not work without DDDs. The compatibility of a device such as a pen, an auto-injector or a pump with a new drug is an important precondition for its success, especially in developed countries. The so-called “pharmerging markets” currently require more basic and affordable technologies. Once

the average unit cost decreases, the optimised DDDs will also be successfully placed in these markets in large numbers. In the future, the trend towards more convenience with a clear focus on patients’ compliance is expected to continue at a smart pace.

Dr Jérôme Freissmuth studied technical chemistry at the Technical University of Vienna, Austria. He holds a doctorate in technical science and has worked as a management consultant focusing on strategy and organizational projects for the pharmaceutical machinery industry and process industry for several years. In 2010, Jérôme Freissmuth joined Robert Bosch GmbH as Director of Business Development, Product Management and Marketing in the Packaging Technology division, Business Unit Pharma. He can be reached at Email:

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Due Diligence for Leachables and Extractables in a Modern GMP Environment Introduction Nearly two decades have passed since extractables and leachables have become a primary consideration for qualifying packaging intended for pharmaceutical use. The complexity of medicines and associated delivery systems continues to evolve, challenging approaches for identifying and quantifying leachables. Regulatory guidance for extractables and leachables exist, but only provide general recommendations for ensuring patient safety. Step-by-step instructions are impractical as dosage form, materials of construction, configurations and dosing are unique for each product; thus, potential impact to the patient must be reviewed case by case. Modern GMP advocates risk-based approaches starting at the drug product development stage, and determining the potential for leachables should be a priority to ensure patient safety. A comprehensive leachable study is a qualitative and quantitative evaluation of constituents that have the potential to migrate from packaging materials into a particular drug product over the proposed shelf-life. Dilemmas remain, including questions of how much testing needs to be done, when and at what sensitivity, and how to determine the process for qualifications. Methodology for extracting contact materials and assessment of the extracts is employed in food, drug and medical device industries, with the common goal of understanding what can migrate from these materials and potentially cause harm. While it is desirable to have definitive methods with identified sensitivity and specifications to fit all cases, it is not realistic; this article will look at extractable and leachable science and discuss approaches, applications and risk-based strategies.


Background Extractables are compounds that can migrate from components of a container closure system when in the presence of a solvent. These extractables have the potential to become leachables, compounds that can migrate into the dosage form from the container closure system over time in storage. It is critical to evaluate components used in the manufacture, containment and shipping of drug products that have direct or indirect contact with the medicine for potential leachables in all types of dosage forms. Several healthcare industry recall cases dating back to 1990 have cited the migration of 2,4,6-tribromoanisole (TBA) and 2,4,6-trichloroanisol (TCA) in tablets due to diffusion and permeation of these molecules from indirect contact with shipping pallets.1 A number of different leachable cases

involving metal cations, tungsten, bromine, butylated hydroxytoluene and organic solvents originating from primary containers and prefilled syringes have been reported in various biologic protein products.2 In order to ensure confidence when moving into scale-up and commercial phases, pharmaceutical manufacturers should identify potential leachable substances during drug development and assess the potential impact to medicinal product and patient. Investigation of packaging components will provide the knowledge needed to establish quality control points throughout the drug product lifecycle that are specific for the componentsâ&#x20AC;&#x2122; application. Unfortunately, the package configurations, manufacturing processes, dosage formulation and dosing - all of which can have an impact on leachables - are not entirely defined

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PACKAGING early in the development process.3 The degree and type of evaluation for each of these components is dependent on many interrelated factors. Many types of materials are used in the pharmaceutical and medical device industry, including those shown in Table 1: Examples of Dosage Forms, Materials and Associated Risks.4,5 All types of materials can contribute to a leachable profile; metal, glass, rubber and plastic can present a risk to patient safety and drug product quality. Particulates have been reported due to leaching or sloughing of constituents from product contact materials, such as glass or roughing in stainless steel, that impacted drug product purity and safety.6 Polymer materials, especially when subjected to heat and solvents, are at risk of migration of constituents from component materials that directly or indirectly contact medicinal products and should be carefully investigated. Science of Migration The likelihood that extractables will result in leachables can be viewed in terms of migrant interactions related to permeation of volatile components into drug products, or accumulation of a migrant at the package product interface and partitioning into drug product or precipitation. The phenomenon of migration encompasses a range of physical and chemical processes that include extractable rates of diffusion, solubility, permeation and chemical structure. The area of exposure will have an effect on the potential for migrating species. Time, temperature and type of contact media/solvents also may affect the rate of migration. In the end, it is the actual exposure over the shelf life of the product that will serve as proof of impact on patient safety because it relates to accumulation of leachables and dosing per day. Although predictive models maybe employed, there are many contributing factors to multicomponent systems in which models can lead to great uncertainty. There are also conditions in which model extraction studies may not capture all potential leachables, or maximum concentrations are not fully represented. Protection of patients relies on properly designed extraction studies followed by full shelf-life

leachable studies. Ingredients used in the manufacture and forming of the materials such as additives, processing aids, residuals and cleaning agents can all be suspected extractables, as well as associated with their potential contamination or degradation of products. A controlled extraction study will provide potential for leachables as a first step. These studies should include extraction after final processing (e.g. sterilisation and assembly) to reveal the actual potential leachables, because this is the system that will be in contact with the drug product over the shelf-life. Migration of additives in polymer materials and impact on food safety have been widely investigated in both the United States and Europe. Legislation has been enacted to protect consumers based on positive lists for indirect food additives. There are examples of regulations that have threshold limits for safety associated with allowable amounts to be added to manufacture the packaging materials and, in some cases, specific migration limits.7,8 While thresholds are important for leachable assessments, there is limited utility for specifications listed in the food additive regulations with respect to qualifying packaging for pharmaceutical use. Migration of additives or chemical constituents from polymers related to the pharmaceutical and medical device industries consider other routes of exposure and impact assessments, which do not necessarily translate to the same type of alerts. Once leachables can be confirmed through appropriately conducted

studies, the impact on the patient and medicinal product can be evaluated. There are many conceivable substances that have the potential to leach, but it is only those that leach above an acceptable level that are of priority. Leachable thresholds have been recommended by the Product Quality Research Institute for orally inhaled and nasal drug products; other dosage forms are still being considered.9,10 The challenge of identifying and prioritising leachables relies on a thorough understanding of packaging component chemistry; however, extractables data alone cannot predict the impact on patient safety. Understanding the chemistry of components and the mechanics of migration will aid in developing suitable extractables and leachables methodology to provide the rationale for control strategies based in science. Extractables can be classified in terms of molecular weight and other aspects to associate probability of extraction by taking into consideration parameters influencing migration. This information is useful to identify appropriate test methodology. The potential safety impact of a given leachable is greatest for those constituents that have evidence of toxicity, but often these occur at trace levels and are not always readily detectable. Classes of potential leachables can indicate potential for a migrantâ&#x20AC;&#x2122;s diffusivity, and surface interaction leading to appropriate investigations of chemical entities should be considered for developing


PACKAGING analytical methodology.11 Influencing factors for potential leachables are referenced in Table 2: Physical Chemical Indicators for Potential Leachables. Extractables Selection of critical components (individual components of manufacture, containment or other material contacting the drug product) and extraction of the test material are the foundations of an extractable study. Adequate component sampling is necessary to establish a comprehensive profile as variability of trace level constituents is inevitable. In addition, suitable sample preparation, material-to-solvent ratio, type of solvent and extraction conditions are critical for acquisition of proper data. The function of the critical components, along with knowledge of the component composition and drug product formulation, should be used to guide solvent selection. Multiple extraction techniques and analytical techniques should be employed.12 The course of the study should be planned to achieve the outcome that will support the study objective and lead to appropriate leachable studies. Extractables assessments have been used to serve multiple purposes, such as overall chemical characterisation, to understand potential for leachables in severe conditions or model systems to represent intended use to indicate probability for leaching under specified conditions. Model systems are intended to mimic products under accelerated conditions, and are typically employed by the food industry and certain medical devices.13,14 Regardless of the conditions chosen for the extraction study, the outcome of the test should provide results that correlate and confirm the type and amount of leachables for pharmaceutical packaging, which is needed to judge patient safety. An example of data generated for a polypropylene sample for the purpose of characterisation compared to that of an accelerated condition to simulate migration is illustrated in Figure 1: Material Characterisation GC/MS Data Compared to GC/MS Data Simulating Actual Use Conditions.15 Conditions for characterisation will 108 INTERNATIONAL PHARMACEUTICAL INDUSTRY

provide information to understand the chemistry of extractables through maximisation of migrants, while a model extraction study employs simulated conditions intended to indicate or predict actual use. It can be realised from the GC/MS chromatograms that the simulated conditions alone would not provide adequate background to establish a comprehensive extractable profiles or leachable assessment; however, they can be a gauge for migration behaviour. The generation of extracts, and identification and measurement of extractables or migrating substances, are time-consuming and should follow a systematic approach of investigation. Understanding the migration behaviour can provide insight into the selection of study parameters and analytical methodology. Polymer characteristics to consider are:16 • Polymer effects (morphology, molecular structure surface energy, entropy) • Physical loss of chemical constituents and conversion to associated degradation products • Chemical reactions within complex mixtures, formation of by-products or interaction products • Migration of chemicals to the surface • Solubility in contact media, precipitation, blooming effect • Extractive loss and mass transport based on molecular motion (diffusion rate or volatilisation) • Outgassing (evaporation of rest constituents) in headspace or accumulation in solution • Permeability or transport of chemicals through a material

Leachables Leachable studies are often not conducted until later in the pharmaceutical development programme. In these studies a drug product is stored in stability under a variety of controlled environmental conditions and analysed for leachables, both qualitatively and quantitatively, at multiple time points over the anticipated shelf-life of the drug product.17 Extractables serve as an indicator of potential leachables, but the probability for migration of chemical entities in a drug product can be unexpected, proceeding slowly and/or under unique, unpredictable conditions. When parameters influencing migration (such as temperature) are accelerated, there are possibilities for anomalies to occur. If conditions or contact media are too mild, it is possible leachables can be missed. Drug product impurity analysis may reveal leachables, but often at high levels, and independent studies are necessary to trace back and correlate to a source. An example of a leachable detected in an impurity assay was demonstrated for an ophthalmic product. Changkang described the methodology used to trace the 0.19 percent impurity back to a lacquer coating.18 To mitigate the risk of chasing abnormalities or missing a critical leachable, targeted studies should be conducted throughout stability while early assessment can warn of potential hazards. Drug stability is a key property to be monitored during development and throughout the drug product lifecycle, but impurity assays cannot always be optimised for suitable concentrations of potential leachables. Drug degradation products, impurities

Figure 1: Material Characterisation GC/MS Data Compared to GC/MS Data Simulating Actual Use Conditions.

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PACKAGING and potential leachables require appropriate methodology for discovery and measurement. A trace level constituent can arise from multiple and/or obscure sources. Leachable methods should be robust enough to capture the expected, the unexpected and potential unknowns. Because methods can be very sensitive, it can be challenging to ascertain the identity of unknowns as well as the source. A substantial amount of time can be invested in isolating and confirming the unknown leachable compound. Comprehensive extractable studies should result in a better understanding of the component chemistry and facilitate the identification of unknown leachable compounds and potential mechanistic pathways. Once the materials in contact with the drug product are understood in terms of their extractables or migrants, potential leachables should be assessed. Decisions are necessary to select which of the observed chemical species may be of concern for patient safety and drug product quality. Leachable studies require several steps before the leachables analysis can begin: compounds should be identified confidently, reference compounds made available for measurements, and appropriate instrumentation and analysis conditions developed, validated and optimised for adequate sensitivity. In addition, the drug product matrix can be anticipated to cause interference with detection techniques and recovery of analytes. Ingredients in drug product formulations often can obscure the response of lowlevel constituents of interest or bind to the matrix, which can also present quandaries. Targeted studies that use optimised/validated methodology for measurement of leachable analytes in the specific matrix offer more accurate information about leachables. The methodology that has been developed for appropriate specificity, recovery and reproducibility of the target analytes within the matrix produces a confident leachables study with correlation to extractables. A â&#x20AC;&#x153;bridgeâ&#x20AC;? between extractables and leachables can be implemented using an approach from a theoretical perspective and/or additional extraction studies under a simulated or stressed 110 INTERNATIONAL PHARMACEUTICAL INDUSTRY

environment. The outcome of this assessment will provide background information to ensure development of appropriate leachable methodology or indicate a potential toxicological concern. These data can have a place in early development, especially when the specific drug product formulation has not been finalised. Solvents that most closely simulate the drug product matrix, placebo or the drug product itself are typically used under accelerated conditions. Often method optimisation is necessary for accelerated studies in complex matrices that may or may not be feasible for leachable development. The full process of material characterisation and understanding extractables through development of specific methodology and monitoring of leachables throughout stability is not a rapid process, but one rooted in Quality by Design (QbD). The goal of QbD is to plan for quality attributes based on a quality target product profile (QTPP). The profile for components in contact with a dosage form should include acceptable levels of leachables. The risk of leachables should be assessed in relation to properties of dosage form, route of administration, patient population, dosing and patient daily exposure. Prior knowledge is useful to guide methodology for discovery and understanding of potential leachables derived from comprehensive extractables evaluations. A planned set of controls should be developed based on leachable studies and a current product and process understanding

to assure product quality with patient safety in mind.19 Conclusion Understanding the science of extractables and correlating to leachables using risk-based strategies will support drug development stages and continue throughout the drug product lifecycle. Application of risk-based strategies will provide rationalisation to influence decisions to assure drug product quality and patient safety related to materials in direct or indirect contact with dosage forms. Final component selection and drug formulation design should include information gathered in material characterisation studies and risk assessments with the aim of decreasing risk later in drug formulation development. Approaches to extractables and leachables studies are varied and dependent on specific products and patient administration. Extracts generated under simulated conditions can be used to bridge the extractables study and subsequent leachables study, but have certain limitations and should not be perceived as a singular leachables study. Selection of target leachables based on risk and development of specific methodology to monitor for leachables over the shelf life of the drug product are consistent with QbD principles. Accurate measurement and assessment of leachables with full method validation is an essential element to protect the quality of the medicine and the safety of patients. Summer 2012 Volume 4 Issue 3



PACKAGING References 1. P  DA Letter, Shipping Pallets: Taint as Clean as They Look, vol. XLVIII, Issue 5, page 30, May 2012: www. 2. M  arkovic, I., Risk Management Strategies for Safety Qualification of Extractables and Leachable Substances in Therapeutic Biologic Protein Products; American Pharmaceutical Review; June 2009 3. B  ak, A. Scientific Risk Assessment Strategies for Managing the Transition from Discovery to Development Drug discovery: AAPS News Magazine, July 2010, http:// newsmagazine 4. C  ontainer Closure Systems for Packaging Human Drugs and Biologics, CMC Documentation CDER/CBER/FDA Guidance for Industry 1999, 5. P  askiet D., “Strategies for Assessment of Leachables in Parenteral Drug Products,” PMP, 2008. 6. M  arkovic, I., CDER/FDA Challenges Associated with Extractable and/ or Leachable s Substances in Therapeutic Biologic Proteins: American Pharmaceutical Review, May, 2007, http://www. americanpharmaceuticalreview. com 7. U  nited Sates Food and Drug Administration, Code of Federal Regulation Title 21 parts 170199, Office of Federal Register National Archives and Records Administration; http://www. 8. E  uropean Communities, Commission Directive 2002/72/EC, Related to Plastic Materials and Articles Intended to be in Contact with Foodstuffs http://ec.europa. eu/food/food/chemicalsafety/ foodcontact/legisl_list 9. P  roduct Quality Research Institute(PQRI) Research Project Proposal: Reporting and Qualification Thresholds for Leachables in Parenteral and Ophthalmic Drug Products: 2007, 10. P  roduct Quality Research Institute (PQRI), Leachables and Extractables Working Group, Safety Thresholds and Best Practices for Extractables 112 INTERNATIONAL PHARMACEUTICAL INDUSTRY

and Leachables in Orally Inhaled and Nasal Drug Products: November, 2006 11. Bart J.C.J., Polymer Additives Analytics, Industrial Practice and Case Studies, Firenze University Press, 2006. 12. Product Quality Research Institute (PQRI), Leachables and Extractables Working Group, Safety Thresholds and Best Practices for and Extractables and Leachables in Orally Inhaled and Nasal Drug Products (OINDP), November 2006, 13. United Sates Food and Drug Administration, Code of Federal Regulation Title 21 parts 170199, Office of Federal Register National Archives and Records Administration; http://www. 14. International Organization for Standardization (ISO) Biological evaluation of medical devices Part 12: Sample preparation and reference materials; and Part 18: Chemical characterization of materials, 15. Hojnicki, P. et al. A Systematic Approach to the Extraction Process, Presented at the PDA Workshop; Extractable Puzzle May 22, 2005. 16. Bart J.C.J., Polymer Additives Analytics, Industrial Practice and Case Studies, Firenze University Press, 2006. 17. Product Quality Research Institute (PQRI), Leachables and Extractables Working Group, Safety Thresholds and best Practices for Extractables and Leachables in Orally Inhaled and Nasal Drug Products, November 2006, 18. Changkang P. et al. Strategy for Identification of Leachables in Packaged Pharmaceutical Liquid Formulations; Journal of Pharmaceutical and Biomedical Analysis, Elsevier B.V, 2007. 19. Prasad Peri, ONDQA/FDA Regulatory Perspectives on Extractables and Leachables, PQRI Workshop on Thresholds and Best Practices for Parenteral and Ophthalmic Drug Products (PODP), Bethesda, MD, Feb 22, 2011 workshops/PODP11/pdfs/PQRI_ Extractables_and_Leachables_ Talk_Prasad_2011.pdf

Diane Paskiet is Director, Scientific Affairs at West Pharmaceutical Services, Inc. Diane has more than 20 years’ experience in polymer analysis relating to product failures, deformulation and migration studies. A sought-after speaker, Diane has authored national and international papers on leachables and extractables and is a PDA Training Institute faculty member. Email:

Laura Stubbs is a senior chemist at West Analytical Services.She authors studyprotocols and analytical reports and serves as the technical contact for clients regarding extractables. Laura 15 years’ experience in the field of extractables and leachables, including managing an organic laboratory group focused on contract extractable and leachable testing of packaging components and drug product, method development and method validation. Email:

Heike Kofler, Manager Technical Customer Service Europe, West Pharmaceutical Services. Dr. Heike Kofler holds a PhD in Life Sciences from the University of Cologne, Germany. She joined West in 2008 as Manager Biotechnology Services in Europe. Since 2011 she is Manager Technical Customer Service Europe and also dedicated to strategic coordination of technical assistance for European start-up companies and universities.

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Labelling and its Role in Pharmaceutical Packaging The ever-increasing legislative demands for patient information on pharmaceutical labelling often poses a packaging challenge to manufacturers and brand-owners. Patient compliance is also a major issue for the industry. Poorly worded, unclear or confusingly presented user instructions can lead to ineffective patient compliance, adverse reactions, side-effects, or at worst fatality. Pharmaceutical labelling goes far beyond functional requirements and fulfilling legal obligations. Millions of dollars are spent each year on developing drugs that are safe for human consumption and that provide effective patient treatment. Yet an estimated 31% of prescriptions that are issued do not get collected, and 40% of those prescriptions will not be taken as directed. The consequences are severe and statistics suggest that the annual cost of non-adherence to the US healthcare system is approximately $US 100 billion, including a staggering 125,000 hospitalisations each year. With these figures in mind, there is an even greater responsibility on manufacturers to ensure that drug usage information is presented to the patient in a manner that is easy to understand and easy to refer to, not just when the drug is first used, but throughout the course. Fortunately, manufacturers are becoming increasingly aware of the importance of labelling on healthcare products in relation to the success rate of a drug. On-pack information speaks directly to the patient, can help to improve userappeal, and most importantly can have a direct effect on patient outcomes. Labelling is an integral part of the patients’ treatment. At a minimum, outside of legislative requirements, for the drug to be effective product labelling must perform the following functions: 114 INTERNATIONAL PHARMACEUTICAL INDUSTRY

•C  ommunication of the brand and type of drug enclosed within the packaging. •C  ommunication of the strength of product, quantity and for whom it is suitable. •U  ser instructions, indications, contraindications and side-effects. However, the self-adhesive label can be used as a powerful tool that goes far beyond the realms of functionality and there is greater scope than ever before for further development. Labelling is a Crucial Method of Communication with the Patient Whilst strict guidelines are in place

to ensure the pharmacist provides the patient with the correct strength and quantity of a prescribed drug, it is extremely difficult under most circumstances to ensure that the patient himself adheres to the prescribed course, without which there is no guarantee of the patient outcome. There can be countless reasons for a patient to stop following the guidelines during a course of medication. Most commonly, a patient will feel better towards the end of a course of medication and therefore no longer think it is necessary to continue. For this reason alone, it is imperative that the guidelines within the labelling Summer 2012 Volume 4 Issue 3



PACKAGING make absolutely clear why a course of medicine must be completed. When deciding upon the design layout of the packaging, consideration must also be given to allow space for the dispensing pharmacistâ&#x20AC;&#x2122;s own label. Often, the pharmacist has no other option than to place his label over existing product labelling and there is a fear that this may obscure important pre-printed data. Once the product leaves the pharmacy, the only form of communication the manufacturer has with the patient is via the packaging. Manufacturers need packaging solutions to deliver cost-effective solutions that satisfy the needs of the industry and consumers: When it comes to designing primary packaging, labelling, informational leaflets, outer packaging and bulk packaging, every element must be taken into account to ensure the packaging line runs efficiently. The fewer packaging elements are involved, the fewer operations there are to go wrong on the packaging line and the less inventory there is to control and reconcile. Combining the primary label and the informational leaflet into a single leaflet-label is one solution. Often there is limited space available to carry all the necessary information. However, compromising the content should not be considered an option. Reducing type size can result in the information being unreadable and may discourage the patient from following guidelines, and this may even contravene legislation. Clearly laid out, easy-to-follow user instruction reduces the need for mid- or posttreatment consultation with a medical professional or pharmacist, and is more likely to result in a positive outcome. In the case of over-the-counter products this may also increase the likelihood of a referral or even repeat purchase, thus having a direct effect on future revenues. Commonly a loose leaflet inserted into a carton is used to fulfil this role, but care must be taken to ensure the user instructions are not instantly discarded by the patient. Alternative options such as multi-page labels are becoming increasingly popular. These may include up to 100 pages 116 INTERNATIONAL PHARMACEUTICAL INDUSTRY

of additional information within the confines of the adhesive label. This has the added advantage of ensuring the patient information is secured firmly to the product or host-container, making it readily available throughout the entire course. Research has shown that multi-page self-adhesive leaflet labels, attached directly to the primary packaging, are generally more likely to be read than a loose leaflet inserted into a carton. More importantly though, they are significantly more likely to be retained by the patient throughout the course of treatment. User-friendly Product Information The way in which the packaging is worded and designed can also have a direct effect on the patient outcome. When providing patient information, the manufacturer must take a number of factors into account to ensure everything is clearly presented within the label and packaging. Literacy and education levels vary around the globe. Children are often dependent upon their parent to administer drugs, but a safe dosage for an adult could be dangerously high for a child. Some patients, particularly elderly ones, may have poor eyesight or find it difficult to remember when and how to take their medication, meaning they have to refer to informational

leaflets repeatedly. Often, patients who are taking several long-term courses of medication at once transcribe basic dosage information onto easy to refer to handwritten reminders. Not only does this suggest that the original information was poorly presented, but it also introduces the possibility for transcription errors and leaves the patient vulnerable should an update to the on-pack information go unnoticed. These are all factors that should be considered when designing labelling. Technical jargon and medical terms that are not commonly understood outside of the industry should be avoided. Whilst the manufacturer cannot be directly responsible for each and every patient, good labelling can improve not just the aesthetic appearance, but also make information easier to follow, thus having a direct effect on patient outcomes. Braille printed labels for the blind and visually impaired are now obligatory within the European Union since October 2010 (Article 56(a) of Council Directive 2001/83/EC). The law states that the name of the medicine along with the appropriate strength must be displayed on all new EU registrations on the label (this includes pharmaceutical products which were launched prior to October 2005). Whilst this is familiar ground for carton manufacturers who Summer 2012 Volume 4 Issue 3



PACKAGING are used to embossing technology, most label printers have taken a little time to catch up. Fortunately, a variety of printing techniques have now been developed to produce high-quality, durable Braille dots directly onto various self-adhesive label substrates, laminates and multi-page leaflet labels. Some of these techniques now surpass traditional embossing methods. Variable Manufacturer On-pack Data Besides the necessary user instruction and guidelines, the label must also include a host of manufacture-specific information. This information is most commonly printed at the time of manufacture either prior to, during or immediately after the label application process. Lot numbers, batch codes and expiry dates must all be included to ensure complete traceability. A variety of self-adhesive label material options are available which allow for high-quality alpha-numeric information to be printed onto the substrate using various printing methods, including thermal transfer, laser oblation and, most commonly, inkjet. Special consideration is needed to ensure such data is durable and cannot be rubbed off, even if products are packed tightly together when shipped. Digital technology means there is now scope to print a range of variable data directly onto labelling and packing to ensure traceability and authentication and to allow for late-stage customisation of product labelling. The demand for this will undoubtedly grow in future and is already being seen on 2D barcodes and variable print code. Such methods of ‘E-pedigree’ printing will allow for even greater control of the whereabouts of products within the supply chain. Clinical Trial Labelling Taking this one step further is the role of the label on clinical trial products. The average cost of a clinical trial is around $800m, so it is crucial that every element of the test runs smoothly and the trial is at no point compromised. For printing companies this is probably one of the most challenging sectors to serve, as the trial must often run within a specific time-frame. 118 INTERNATIONAL PHARMACEUTICAL INDUSTRY

Commonly, trial participants are widely spread geographically and therefore labelling is required in more than one language, increasing the demand for yet more copy space. Also, there must be total confidence that printed information stays with the drug throughout. Again, leaflet-labels are increasingly being used to meet these dual challenges. Quality and packaging consistency must be at an even higher level than normal, making clinical trial printing a specialist discipline. A minor printing defect on the label such as a fleck or hickey can potentially lead the medical practitioner involved to falsely believe that they are administering the placebo or the real drug, and this could unintentionally cause them to skew results. There is a range of labelling features and options that may be used to assist both the medic and the patient during a clinical trial. Unprinted portions for overcoding can allow additional information to be added to the label and late-stage customisation. Detachable portions may be included on the label for preprinted or handwritten information that can be added to patient notes for reference at a later date. Packaging Security and Integrity Brand protection is a growing concern in all industries, and nowhere more so than in the pharmaceutical sector. According to CHP Packer International, counterfeit medicines cost the global economy an estimated $1,000bn annually. They not only undermine trust and integrity in brands, but can cause illness or even fatalities amongst the unsuspecting public. Even the rumour of a counterfeiting issue can devastate the value of a brand overnight. Advancements in label printing technology are playing an increasingly important role in combating the counterfeiters. Holograms and tamper-evident strips provide overt identification on products. This is particularly useful where over-thecounter medicines are concerned. Additionally, a wide range of covert options are now available, especially for high-value drugs. Hidden messages, images and even unique codes may be printed directly onto label substrates for quick identification

within the supply chain. In fact, technology advancements mean that the manufacturer is now able to encrypt artwork with their own integrity marks and commission the printed labels without the printer even being aware of this information. Clearly there is a cost attached to such advanced technology, and this does not suit all product applications. Manufacturers need to be mindful of the public’s growing awareness of counterfeiting issues, and the added value that overt anti-counterfeiting features can bring, but must also differentiate between adding value and simply adding cost. Final Thoughts: Given the huge cost of developing, testing and launching a new drug, it would be a false economy to use cheaper and potentially less effective packaging if doing so risks compromising patient compliance and therefore the effectiveness of the drug. There is undoubtedly more scope for improved labelling and packaging than ever before, even within markets that are not so heavily led by legislation. The growing realisation is that a drug is only as good as the usage information conveyed to the patient. Manufacturers no longer treat packaging as an ‘afterthought’ and recognise the need for clearly-presented, easy-to-follow packaging that is conducive to positive patient outcomes.

Steve Moore is Client Support Executive at Fix-a-Form International Ltd - a UK based leaflet label specialist and licensor to the world’s largest network of leaflet label printing companies. The Fix-a-Form network comprises 20+ independent label printing companies providing local manufacture across the globe. Visit: for more information or Email:

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Bag-on-Valve â&#x20AC;&#x201C; Optimal Solution for Pharma Products With the total separation of product and propellant, a bag with superior barrier properties, and complete oxygen protection, Bagon-Valve is the optimal solution for pharmaceutical and health products. Many products can be made in a sterile presentation. Sterility is guaranteed â&#x20AC;&#x201C; even when the can has been opened and used frequently over a long period of time. With tax-based healthcare services burdened by increasingly higher costs for pharmaceuticals and the evermore stringent regulation governing the introduction of a new drug on the market, there is a growing trend moving from prescription (Rx) to nonprescription, over-the-counter (OTC) products. It has turned out that the Bag-on-Valve (BoV) may in several cases be a suitable technology for products to make this switch, as marketers are looking for modern, innovative packaging that can attract consumers. Products for which the pharmacological claims are not the primary function can often be classified and registered as medical devices utilising the BoV technology. The physical or mechanical function of a medical device spray product is suitable for this modern aerosol system. Being classified as a medical device, rather than pharmaceutical, a product can reach the market much faster, and in the EU, an application for registration can be made once instead of the more complicated and costly drug application using the centralised, decentralised or mutualrecognition procedures. Registering an OTC-product as a medical device can provide a more cost-effective way of entering the European market. There is therefore an increased interest in giving old products new, more convenient applications as well as finding niches for totally new 120 INTERNATIONAL PHARMACEUTICAL INDUSTRY

products using the BoV technology for administration. Originating in the cosmetics industry, BoV technology has entered into the pharma industry where it is expanding, and now covers saline/seawater nasal sprays, wound cleansing/irrigation sprays, eyewash sprays, dermal drug delivery products, disinfectant sprays and veterinary products, among others. Convenient and Environmentallyfriendly The Bag-on-Valve consists of a rolled-up multi-layered flexible pouch attached to an aerosol valve. In the production of a BoV product, compressed air or nitrogen is charged into the can, and an aerosol valve with the bag attached is crimped onto the container. The formulation is forced through the aerosol valve stem to fill

the bag. When the spray button is pressed, the product is squeezed out of the bag by the compressed air/ nitrogen, which creates the dispensing as a spray, cream or gel (see graphic for details). As such, BoV is an ideal barrier pack for liquids, gels, creams and lotions, keeping high product integrity and protection against oxygen. Hence, for many formulations the amount of preservatives needed is very low and may in some cases be excluded altogether. For sterile products it is possible to arrange aseptic filling or use gamma irradiation to achieve sterility. The propellants eliminate the need for pumping out the content, which often is a great advantage compared to, for example, pump sprays or trigger sprays. Instead the content can be sprayed, smoothly and evenly, giving the user perfect control Summer 2012 Volume 4 Issue 3



PACKAGING over the administration of the product. Another major convenience connected with BoV is the fact that it is possible to spray from any angle without losing pace or force in the spray. The degree of emptying is very high, up to nearly 100 per cent for water-based solutions. If desired, a Bag-on-Valve aerosol can produce a smooth and even layer of a film-forming product or a protective barrier cream to the skin – without any need for additional handspreading, thus resulting in advantages in hygiene and patient integrity. This means that the product can be applied without actually touching the skin, avoiding the spread of contamination from the hands, ideal for sensitive product applications. For healthcare professionals and patients alike it is less encumbering having someone to spray rather than rub the content onto the skin. Since a Bag-on-Valve aerosol functions equally well at all angles it is easy to use – even in emergency situations. A Wide Range of Applications BoV can be used to upgrade the packaging of a product, increase the marketing opportunities, and/or to add new convenience to the formulation, for instance turning an emollient cream into an emollient spray. For an innovative pharmaceutical company it may be interesting how they can challenge established brands simply by using an alternative spraying technology. With its many dispensing possibilities, Bag-on-Valve is suitable for direct dispensing even in such sensitive areas as eyes or ears – even in children. One of the most common formulations used in BoV medical devices is different saline solutions. They are used in several different applications, such as nasal sprays, wound cleansers, ear-cleansing sprays and also eyewashes. Market leading nasal spray brands like Simply Saline (US) and Stérimar (EU), both from Church & Dwight Co. Inc. are wellknown examples. Nasal sprays in BoV are widely used based on saline or natural seawater solutions. Wound care on the other hand is rather “virgin soil” when it comes to BoV, even if wound cleanser with saline 122 INTERNATIONAL PHARMACEUTICAL INDUSTRY

Bag-on-Valve Technology 1. The Valve Bag-on-Valve systems are available with both female and male valves. 2. The Bag FDA-approved laminated bag (three or four layers depending on product requirements). Available in a large range of sizes. 3. The Product Suitable for liquid as well as viscous products, such as gels, creams and ointments, in a number of application areas.Optimised for oxygen-sensitive and sterile products. 4. The Propellant Because of the separation between product and propellant, Bag-on-Valve can be used with compressed air or nitrogen. 5. The Actuator A large range of standard actuators depending on product demands. 6. The Can Standard aerosol can, aluminium or tinplate. All sorts of shaped cans can be used. No limitation in marketing solutions. The Cap – Can be used with many kinds of standard overcaps.

STEP 1. Placing of valve and bag inside the can

STEP 2. Gassing (air/ nitrogen) followed by crimping of the valve onto the can

STEP 3. Pressure control

STEP 4. Product filling in the bag followed by weight control

STEP 5. Actuator and cap placement



PACKAGING solutions for hospital and community care have been on the market for more than 10 years in the EU and the US. There is a lot of product development currently in process in this segment. One example of an innovative approach using BoV is the new product Granulox (SastoMed, Germany) for treatment of chronic wounds. The Granulox product uses haemoglobin in order to improve oxygen supply to chronic wounds. The haemoglobin-based formulation sprayed onto the wound area is able to penetrate the exudate and thereby increase the level of oxygen in the wound. The wound situation improves and wound healing is accelerated. Wounds stagnant due to hypoxia can also be activated. Several companies are also recognising the benefits of BoV for the application of silicon-based products. An example of this is the growing market for medical adhesive remover sprays where sophisticated silicone formulations are sprayed to gently and painlessly remove adhesive products such as stoma pouches and wound dressings. Conventional adhesive dressing removal is often difficult for the healthcare professional and unpleasant for the patient. Another upcoming area currently catching interest is throat spray, with the aim of soothing and preventing sore throat symptoms during colds and allergies. Maybe we will see a similar range of throat spray products on the market as is already available for nasal application. In the ENT segment there are also BoV-products on the market that help to prevent build-up of earwax. Occupational safety is another area where BoV products are increasing their market shares. A BoV first aid eyewash, such as is available on the EU market, is a good complement to the water pipe or gravity-fed eyewashes that are usually found at industrial workplaces. Benefits include a lesser amount of liquid that lasts longer, portability and the ability to be either personal or available in a wall station. In products such as first aid eyewashes, the effective flushing time has to be as long as possible. This can be achieved with very fine spray actuators and a controlled discharge flow rate. Thanks to the separation of 124 INTERNATIONAL PHARMACEUTICAL INDUSTRY

Fact box: Advantages of Bag-on-Valve (BoV) The Bag-on-Valve system offers major advantages for a range of aerosol products where safe and contact-free dispensing is required. Bag-on-Valve gives new marketing opportunities through improved consumer conveniences, product protection, and controlled discharge of the product. Safety and environmental benefits •N  o need for flammable propellants • Hygienic and sterilisable •U  sed with eco-friendly air or nitrogen • Less need for preservatives Consumer benefits • Up to 100% product emptying •L  onger shelf-life with less preservatives •E  ven and controlled spraying pattern for optimal result • No pumping motion needed • Can be used at all angles • Reduced spray noise • Less-chilling product discharge

product and propellant, Bag-on-Valve offers a more constant pressure and flow rate until the last drop. Let’s not forget disinfection products used in critical environments like pharmaceutical industry cleanrooms. Double-bagged sterile disinfection sprays for this purpose with isopropanol alcohol or ethanol are now available using the BoV packaging. The greatest convenience of this is that they are easily sprayed on machinery and surfaces in the critical areas. Integrity of Brands BoV products for the pharmaceutical market are often manufactured by contract manufacturing organisations (CMOs). For sterile products, some CMOs of BoV can offer aseptic production. A more common method to realise sterile products is the use of gamma irradiation of the final product. The manufacturers may be focusing on GMP production for products classified as drugs or on production of medical device classified products based on

Production and distribution benefits •L  onger shelf-life for oxygensensitive products • Effective filling process •S  uitable for both liquid and viscous products •C  an be used with standard actuators and aerosol cans.

the ISO 13485:2003 standard. Some CMOs in the EU, such as Aurena Laboratories, offer medical device products, developed in-house, for companies to sell and distribute under their own brands. In these cases the final products are already registered and CE-marked according to the European Medical Device Directive. The brand name and artwork design of the new private label will be added to the technical file (dossier) of the registered product and become a part of the registered product. The procedures will be different depending on the classification of the product. Depending on the type of contractual arrangement between the CMO and the brand owner it can be either the Summer 2012 Volume 4 Issue 3

PACKAGING CMO or the brand owner that is stated as the legal manufacturer. This can ensure total integrity of the brand, as the production source is not disclosed on the packaging. With BoV the cost per ml can often be decreased, particularly regarding nasal sprays with a filling volume of 50ml and upwards, compared to pump sprays that normally contain a lesser filling volume. Available standard BoV packaging covers a broad range from 30 to 500ml. The components for BoV are made by other suppliers, of which Aptar Group, Lindal Group and Coster are among the leading companies. Special aerosol filling machines are needed to fill BoV products.

Leading suppliers of filling machines are Pamasol (Switzerland) and MBC Aerosol (USA). For transportation, the BoV products are classified as aerosols (UN 1950). For the pharmaceutical market the cans are most often made of aluminium, even if tinplate BoV systems are also available. The cans are also light to carry and normally fit into the recycle programmes established in most countries. The cans also offer interesting marketing opportunities, since they represent quite a large display area in the store and can be designed in a highly attractive way, giving the brand name a high degree of exposure.

Magnus Hedman Mr Magnus Hedman is the Co-founder, Partner, Marketing Director, Business Development Manager of Aurena Laboratories AB. He is a seasoned entrepreneur and business development specialist who has been involved in the medical device and pharma industry for over 20 years. International Sales & Marketing Manager for Swedish respiratory care company Aiolos Medical, 1990 to 2000. He co-founded Aurena Laboratories in the year 2000 and has developed a range of medical devices in the Bag-on-Valve packaging for the Pharma industry. He has led the dedicated work to develop Aurena into a leading international company of medical device aerosols. Email:



Planning and Performing Clinical Trials in Children and Adolescents Jaypreet Dhillon: We are receiving an increasing number of queries from our readers asking us to publish more information relating to running paediatric clinical trials. Is this a phenomenon that you as a centralised laboratory are aware of? Hermann Schulz: Indeed, the current regulatory climate and the need to meet FDA and EMA paediatric regulations has increased the burden on pharmaceutical companies wanting to register new medications. The development of paediatric investigation plans (PIP) requires that scientists writing clinical protocols take the needs of children in the various age groups into consideration.1 JD: You refer to the age groups. Can you provide more details? HS: When talking about children we should have in mind that “children” is not just a homogenous population. The various age groups significantly differ in their organ maturity and metabolic patterns, and therefore will respond unequally to drug treatment depending on the organ functions affected by the drug’s pharmacokinetic and dynamic effects. When planning study protocols and study drug production, the galenic formulation of the study medication plays a significant role. In summary, paediatric trials require a more individualised approach to study planning and performance when compared to studies in the adult population (also refer to Table 1).

Unfortunately, we should be aware that children are not just small adults. As children significantly differ from adults in their metabolism patterns they consequently should not be treated with drugs simply by extrapolating dosing information developed for adults. This therapeutic approach followed so far may cause a number of problems, such as a possible lack of efficacy due to underdosing or toxic effects due to overdosing, or just no therapeutic effect at all. With regard to health risks associated to over- or underdosing, off-label use of drugs in the paediatric environment should be avoided. JD: When did regulatory implement these changes?


HS: To improve the health of children, both the US FDA (BPCA Best Pharmaceuticals for Children Act and PREA Paediatric Research Equity Act) and the European Parliament implemented appropriate regulations in 2002 and 2007, respectively. The authorities’ goal was and still is to increase the involvement of children when developing new medications. JD: The administrative logistics behind a clinical trial should not be much different between a study in adults and a study with children, should they?

JD: Why are paediatric trials coming into focus now?

HS: Well, the logistics involved in collecting and handling blood samples in paediatric trials do significantly differ from the standards used in “adult” studies. And this is the reason why central laboratories are playing an increasing role in this new type of trial.

HS: Unfortunately, it has been and still is standard medical practice to treat children with drugs that have been approved for use in adults only. In the past, clinical trials did not include children and children were not considered to be a target population when planning clinical trials.

JD: Is there a way for pharma companies to avoid running paediatric studies and hence avoid this extra workload and costs? HS:This is an important issue. Yes, the requirement to submit a PIP may be waived for specific medicinal products or classes of medicinal products that


are likely to be ineffective or unsafe in one or more paediatric age groups, that are intended for conditions that occur only in the adult population, or that do not present a significant therapeutic benefit to paediatric patients compared to existing therapies. JD: Let me return to the more practical issues. Embarking on paediatric clinical trials also means additional headache in obtaining informed consent. HS: Participation of a child in a clinical trial follows the same rules as adults. These rules are based on an inclusion/ exclusion criteria. Regulation and laws foresee additional safeguards when children are participating in clinical trials to ensure the safe and ethical treatment of this population. The standards for obtaining parental consent for a child to participate in a clinical trial vary geographically. While consent can be given at the age of 16 in one country, this may only be possible in another country if the adolescent is over 21 years of age. Children below this age limit may give their assent. JD: Is there a difference between consent and assent? HS: Granting “assent” expresses a willingness to participate in research by persons who are too young (as defined by law or regulation) to give informed consent, but who are old enough to generally understand the proposed research, the expected risks and possible benefits, and the activities expected of them as study participants. If assent is granted, informed consent must still be obtained from the child’s parents or guardian. JD: I can imagine that collecting blood samples from lactates or small children brings limitation to the study design with respect to the number of control visits and blood collections. HS: Absolutely. The most challenging Summer 2012 Volume 4 Issue 3

INTERVIEW aspect of running clinical trials in children below two or three years is their overall reduced total blood volume. Consequently, when treating children, blood cannot be drawn in the same quantities or using the same type of tubes as used for adults. It is therefore critical to identify appropriate sampling techniques. The critical questions are a) how much blood can be collected, and b) which sampling techniques are available. JD: But what does this mean for practical purposes? How much blood can be withdrawn with minimal risk to children? HS: Institutional review boards (IRBs) or ethical review committees tend to consider a single blood draw equivalent to 1-2% of the child’s total blood volume as minimal risk. But one blood draw is generally not sufficient when running a clinical trial. Therefore, other experts define it as safe if the cumulative blood volume collected over an eight-week period does not exceed ten per cent of the child’s total blood volume. Unfortunately literature about this item is not consistent. JD: Can you give an example? HS: Most publications recommend the total blood volume to be drawn in a 24-hour period to be below 3-5% of the total blood volume. This would mean approximately 10ml blood for a newborn child with an average weight of 3kg and a total blood volume of 270ml, or approximately 3ml in case of a premature baby with less than 1kg body weight and a total blood volume of up to 90ml. JD: I guess the individual medical situation must be taken into account, too?

the samples are easy to obtain from finger, ear lobe or heel-prick. Suitable commercial sampling papers absorb the blood sample and distribute evenly through the paper to leave a spot of blood which is allowed to dry in situ. Using the DBS technology, typical sample size is approx. 15µl. Thus, this method could help to overcome the challenge of collecting multiple samples to perform pharmacokinetic and pharmacodynamic evaluations which are also required in paediatric studies. With its well-characterised advantage of low sample volume and the relatively non-invasive nature of the DBS sampling method could be ideally suited for this type of clinical trials. JD: But with DBS a laboratory must use different analytical equipment. HS: Absolutely. There are a number of techniques that can offer significant reductions in the volumes of biological fluids required for each analysis e.g. multiplexing techniques such as Luminex. This method not only reduces the sample volume required, but also improves the efficiency when compared to single analyte methods. This technique allows performing analysis of a large number of analytes on extremely small volumes of approximately 50µl per sample. Another assay using nanotechnology for quantitative assays of macromolecules is Gyrolab. Up to five assays can be run simultaneously on Gyrolab on the same sample. Sample size for this method is as little as 10µl.

HS: Correct! JD: Is there a way to avoid collecting blood from babies in the ml-range? HS: An alternative sampling method to the conventional blood withdrawal is Dried Blood Spot (DBS; 2). DBS offers significant practical advantages over traditional sampling methods, as

JD: One side of the coin is collecting low blood volumes; the other side is to find a laboratory able to handle such minimal blood volumes. HS: My words. This is of special interest, because most automatic analyzers have a so called “dead volume” which means that the vial

which is inserted into the analyzer has to contain more fluid than actually needed for the determination itself. Therefore before planning a paediatric clinical trial an appropriate laboratory should be found which is able not only to run the methods needed, but also to show sufficient expertise in supporting paediatric studies and in handling microvolumes. JD: Thank you very much, Hermann, for this brilliant insight. References 1. K . Neuer-Etscheidt, H. Schulz, “Clinical trials in children challenge central laboratories”, Journal for Clinical Studies Vol. 3 Issue 1: 1619, January 2011 2. ICH Guidance E11: “Note for Guidance on Clinical Investigation of Medicinal Products in the Paediatric Population” (CPMP/ICH/2711/99) human/ich/271199en.pdf

Before founding INTERLAB in Munich in 1994, Dr Hermann Schulz held senior R&D positions in the pharmaceutical industry (Merck&Co, Astra-Zeneca/ICI and UCB/Schwarz) over 12 years. As a visiting professor, Hermann is head lecturer for applied clinical pharmacology in the postgraduate course “Pharmaceutical Medicine” at the University DuisburgEssen (formerly Witten-Herdecke). He has written more than 35 scientific publications and is invited as speaker to international conferences such as DIA or IIR. Email:



11th ScanBalt Forum Features Biomaterials and Russian Collaboration Welcome to ScanBalt Forum 2012! The annual ScanBalt Forum enters its second decade on 20-23 November in Tampere, Finland. This year, the life science community on top of Europe will gather in Tampere Hall, Scandinavia’s largest congress and concert centre, to exchange ideas and discuss the current state and future prospects of ScanBalt BioRegion. As ever, ScanBalt Forum brings you an extensive range of topical issues in biotechnology and life sciences discussed by key figures in the field representing the 11 countries of ScanBalt BioRegion. ScanBalt Forum 2012 is hosted by the Baltic Institute of Finland, Tampere University of Technology, Department of Biomedical Engineering, and BioMediTech. Innovation and Collaboration ScanBalt is the organisation for the Baltic Sea or Nordic-Baltic Region’s Health and Life science community, named ScanBalt BioRegion – a strong brand supporting regional promotion and the common benefit of Europe’s most innovative macro-region, the Baltic Sea Region. ScanBalt BioRegion comprises over 1300 research institutes, public institutions, universities, and hospitals, as well as over 2600 businesses in Denmark, Estonia, Finland, Iceland, Latvia, Lithuania, Norway, Poland, Sweden, the northern part of Germany, and the north-western part of Russia. The region of Northern Netherland is affiliated to ScanBalt BioRegion. It is the mission of ScanBalt to establish ScanBalt BioRegion as a preeminent, globally competitive macroregion and innovation market for health and life sciences through investments in smart growth, sustainable development, and specialisation. From this underpinning, ScanBalt Forum emerges as a tool for the bottom-up initiation of transnational cooperation, 128 INTERNATIONAL PHARMACEUTICAL INDUSTRY

for the bridging of academic research and industry and, ultimately, the strengthening of regional cohesion and cluster development. For diverse organisations from business and industry to research and public policy, the annual ScanBalt Forum provides an invaluable platform for collaboration and international networking, fostering the innovation and competitiveness of the region. From Biomaterials to the Russian Market Discussion at this year’s ScanBalt Forum is set to encompass, for instance: -T  he coming EU Programming Period 2014–2020 and the EU Strategy for the Baltic Sea Region -S  canBalt Health Region – the Health Flagship in the EUSBSR -B  iomaterials and Tissue Engineering Days, and -B  ioLifeScience cluster collaboration with Russia. Biomaterials and Tissue Engineering Days are organised in connection with this year’s ScanBalt Forum. Constituting a Finnish strong suit, biomaterials and tissue engineering will provide a fine access point into life sciences in Finland as well as the wider ScanBalt BioRegion. The focus of Biomaterials Days is to strengthen the cooperation in the field of biomaterials, tissue engineering, and biomedical technology research and to discuss possible industrial applications for the future. Another central topic in this year’s Forum is cluster collaboration with Russia, where a life science market with tremendous potential for future growth and for new success stories has been emerging for some time, giving rise to a keen interest in cooperation in terms of both business and research. Europe and the Baltic Sea Region are poised to make the most of the new opportunities on offer, and this

year’s ScanBalt Forum addresses that prospect in earnest in Tampere, an active region for Russian collaboration in Finland. BioLifeScience cluster collaboration with Russia has been designated as one of the key topics of this year’s Forum, and, as such, the issue will of course feature prominently in the conference agenda. High-level guests such as representatives of the Ministry for Economic Development of the Russian Federation and of the City of St. Petersburg are expected to give speeches in plenary sessions and in workshops tailored for the further development of the Baltic Sea-to-Russia and, moreover, ScanBalt BioRegionto-Russia collaboration. The occasion could not be more suitable for a lively exchange of ideas, for discussion on the current state and future prospects of ScanBalt BioRegion, and for the promotion and advancement of Russian collaboration. Tampere – A Centre of Life Science The City of Tampere lies at the heart of Finland’s second most significant economic region. The largest inland centre in the Nordic countries, Tampere is the beacon of scientific, cultural, commercial, and industrial progress in Finland. Tampere is the fastest-growing life science centre in Finland. The city has a combination of multidisciplinary technological, biomedical, and medical expertise in education, research, healthcare, and business sectors. Indeed in recent years, the health, wellness, and biotechnology sector in the Tampere region has been the fastest growing in Finland and received the largest number of private investments in business development. The region’s wellness and health technology cluster provides some 30,000 jobs. Further information and registration for ScanBalt Forum at: Summer 2012 Volume 4 Issue 3




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Galleon announces positive second phase I study results of GAL-021 to treat respiratory depression Galleon Pharmaceuticals, a leader in the development of novel medicines to treat breathing control disorders, has reported positive results from a second phase I clinical study of its investigational drug GAL-021 confirmed previous findings that the drug has dose-dependent effects on respiration. The purpose of the new study was to evaluate higher doses of GAL-021 to determine an appropriate dose range for future studies, including an upcoming phase I proof-of-concept study utilizing conditions simulating post-operative patients who have impaired respiration. The new study evaluated a single dose of GAL-021 in 18 healthy volunteers using higher doses of GAL021 than the previous phase I trial. The placebo-controlled, double-blind, crossover study confirmed the drug’s positive effects, including improved minute ventilation (a capacity measure of air supplied to the lungs) and decreased end-tidal carbon dioxide levels (a measure of the effectiveness of alveolar gas exchange), demonstrating a clear dose response on these respiratory parameters. Side effects of the drug were generally mild to moderate at the highest dose in some subjects; the improved lung ventilation was sufficient to increase in minute ventilation by more than 50 per cent and in parallel lower CO2 levels more than 25 per cent. “This study confirmed our previous phase I findings and establishes a clear dose range for future studies,” said James F McLeod, MD, Galleon’s senior vice president, clinical research and development, and chief medical officer. “We identified a maximum tolerated dose of the drug in healthy volunteers which produced hyperventilation and decreased CO2 levels, and can now proceed with plans to evaluate the effects of GAL021 in those with opioid-induced respiratory compromise. Our goal will be to demonstrate that GAL-021 134 INTERNATIONAL PHARMACEUTICAL INDUSTRY

can reduce or prevent respiratory depression associated with opioid use following surgery or other medical procedures.” GAL-021 is a proprietary small molecule delivered by intravenous administration to treat or prevent acute respiratory insufficiency in surgical and critical-care patients following the use of anaesthetic, analgesic and sedative drugs individually or in combination. Galleon is the first pharmaceutical company to focus on the medical treatment of breathing-control disorders associated with surgical and procedural sedation, chronic pain management and sleep apnea. Source: IPI Staff Reporter, Cecilia Stroe FDA Approves Inhalation Aerosol For Asthma. The Boston Globe reports in its “Business Updates,” that Acton Pharmaceuticals announced Thursday that the Food and Drug Administration has “approved its supplemental new drug application for Aerospan Inhalation Aerosol [flunisolide HFA, 80 mcg], an asthma treatment for patients 6 years of age and older.” The Marlborough, Massachusetts-based company “said it is now planning to launch Aerospan in early 2013 into the $8.3 billion US inhaled steroid market.” In the announcement, Acton noted that asthma “accounts for an estimated $11.5 billion” in healthcare costs, annually. The Worcester (MA) Telegram & Gazette (9/21, Eckelbecker) adds that Acton said that the FDA’s approval of Aerospan Inhalation Aerosol “as a preventive treatment” was based on the results of clinical trials comprising “more than 1,700 people.” The Telegram & Gazette notes that Acton “acquired the rights to Aerospan under a license from Forest Laboratories Inc. in 2009.” Source: IPI Staff Reporter, Cecilia Stroe

Cresset Unveils sparkV10 – the Update to FieldStere, a Leading Bioisostere Software Tool Enabling Medicinal and Computational Chemists to Find New Structures in New Chemical Space Cresset, an innovative provider of software and services for molecule designers, unveils sparkV10, the next generation of its bioisostere software tool FieldStere. Building on FieldStere’s industry-leading ability to find structurally novel biologically equivalent replacements for key moieties in molecules, this update provides powerful new capabilities including Cresset’s new XED3 force field, new clustering algorithms and access to ADMET and physicochemical property predictions on bioisosteric replacements. Medicinal and computational chemists use sparkV10 to generate novel and diverse active compounds by replacing sections of existing molecules with bioisosteric fragments. The results are used to create or improve IP positions and optimize leads while minimizing ADME and toxicity issues. sparkV10 is available for deployment on standard desktop computers and can be integrated into workflow technologies. sparkV10 will:Generate highly innovative ideas for lead molecules in new areas of chemical space, overcoming the ‘chemotype trap’. Grow fragments to generate novel, drug-like molecules. Filter results to find the bioisostere with the right mix of physicochemical properties and biological activity. Tailor results by selecting the chemistry allowed for the replacement moieties. Display results in detail side-byside and cluster similar chemical scaffolds. Search for moieties from real, published or unexplored compound databases or create custom databases based on proprietary chemistry. Cresset CEO, Dr Robert Scoffin said, “This update to sparkV10 reflects Summer 2012 Volume 4 Issue 3


the new names of the molecule design tools in our Next Generation Chemistry Software portfolio. sparkV10 builds on the excellent results of FieldStere to give companies improved IP positions by patent-busting or patentexpansion as well as benefiting from lower-risk and lower-cost strategies for the discovery of new chemical entities. Additionally, by exploring bioisosteric replacements and using field-similar rather than structure-similar molecules, sparkV10 generates new leads, even in crowded therapeutic areas.” Scientists interested in Cresset’s Next Generation Chemistry Software can download free demos from the company’s website. Meeting the needs of computational and medicinal chemists, Cresset’s new suite of Next Generation Chemistry Software comprises: forgeV10 a powerful computational suite to understand SAR and design; torchV10 an intuitive design and 3D SAR tool for medicinal chemists; torchV10lite a free 3D molecule viewing, editing and drawing tool; blazeV10 an amazing ligand based virtual screening tool; sparkV10 an exciting and powerful way of generating novel and diverse structures. Source: IPI Staff Reporter, Cecilia Stroe Staying at the forefront of optical metrology Setting a new standard of performance in measuring technology, Olympus has released an updated version of its LEXT OLS4000 confocal laser scanning microscope (CLSM) software. Compatible with Windows 7, 64 bit operation, the LEXT software version 2.2 allows users to capture high resolution images up to 10 times faster than the previous version. In addition, the new 3D multilayer function can measure transparent layers, allowing the accurate measurement and subsequent analysis of multiple layers within a single sample. As a result, users can obtain 3D images to accurately assess surface metrology with ease and accuracy.

When imaging surface roughness on the micrometre scale, high resolution is essential to obtaining accurate and meaningful data for analysis. The CLSM design of the LEXT OLS4000 facilitates non-contact measurement of surface roughness, thus maintaining sample integrity while obtaining the dimensions of micro-geometries, at high resolution. The recently updated software even enables these high resolution measurements to be obtained up to 10 times faster than before. As such, users can complete their measurements in a time-efficient manner, allowing them to focus on the analysis and applications of their findings. The incorporation of a multilayer mode enables users to quickly and easily recognise the peaks of reflected light intensities on multiple layers within a sample – for example users can observe and measure the upper surface of a transparent specimen as well as measuring the thickness of each individual layer. The shape and roughness of a layer sitting below transparent material, such as a metal surface sitting under a resin layer, can therefore be measured with ease. The Olympus LEXT OLS4000 has a large and very fast MEMS scanning mirror along with dual pinholes. The larger mirror enables the system to provide superior optical quality, and the increased scan speed reduces the time it takes to create a 3D image of a sample. The 405 nm laser and dual pinholes operate simultaneously to ensure that the system provides the highest possible resolution and contrast, as well as enabling steep slope detection up to 85º, ensuring that even the most complex of surface topographies can be imaged and analysed. These features, along with the accurate Z-drive and stage, combine seamlessly with the advanced yet intuitive, new software. The measurement capabilities and flexibility required are therefore provided for even the busiest of laboratories Source: IPI Staff Reporter, Cecilia Stroe

TTP Labtech Supports Drug Discovery Researchers at MipTec Experience the mosquito HTS and acumen eX3 systems at booth D29 TTP Labtech will be offering delegates at MipTec 2012 the opportunity to experience their innovative automated solutions for effortless drug discovery first-hand. The flexible mosquito® HTS liquid handling system for highthroughput screening capabilities, and acumen eX3 laser scanner for rapid whole well high content imaging, will be demonstrated at booth number D29. TTP Labtech will also be supporting researchers with two ‘FREE to register’ User Group Meetings at the event, on High Content Screening and Biobanking, 26th and 27th September respectively. Delegates can also learn more about the applications of the systems in a number of poster presentations. The acumen eX3’s unrivalled imaging capabilities for faster than ever cell based screening are discussed in the poster ‘Rapid profiling of multiple toxicity indicators using a laser scanning imaging cytometer’. While, the renowned precision and accuracy of the mosquito portfolio for rapid, cost-effective dispensing are highlighted in the posters ‘Robust nanoliter liquid handling for flexible assay miniaturisation’ and ‘Bridging the gap for automated low volume liquid handling’. Source: TTP Labtech



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Aesica Almac Group Analytical Biochemical Laboratory BV Aurena Laboratories AB BARC Global Central Laboratory Berlinger & Co. AG BENEO-Palatinit BioCon Valley BioM Biotech Cluster Development GmbH Bioneer A/S BioPartnering Future Europe BIO-Europe 2012 Biotech Services International Ltd Biowin Bobst Group SA Catenion GmbH Cellgenix GmbH Centrical Global Ltd. Colder Products Company Colep Bad Schmiedeberg GmbH Dales Pharmaceuticals Dividella AG Envirotainer AB ERT Excard Research GmbH ExpreS2ion Biotechnologies Extedo FeF Chemicals A/S Flexible Medical Packaging Ltd. Forresters Fujifilm Diosynth Biotechnologies Glycotope Biotechnology GmbH Health - Made in Germany Health Protection Agency InGell Labs BV ISCE Expo – 2012 (UBM Plc) CPhI India – 2012 (UBM Plc) LC Patents LI-COR LISA - Austria Wirtschaftsservice Gesellschaft mbH LTS Lohmann Therapie-Systeme AG LYOFAL S.A.S. Miller Insurance Services MPI Research One2One – A service of Hospira Orbsen Consulting Patheon Inc. Phage Consultants Pharma Publications Pöppelmann GmbH & Co. KG Qualogy Ltd Rapid Micro Biosystems Europe GmbH Sanbe Farma Scottish Development International Sofrigam SynCo Bio Partners TOSOH BIOSCIENCE GmbH Vindon Scientific West Pharmaceutical Services Woodley Equipment Company Ltd.

Summer 2012 Volume 4 Issue 3

Profile for Pharma Publications

International Pharmaceutical Industry  

International Pharmaceutical Industry

International Pharmaceutical Industry  

International Pharmaceutical Industry

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