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Advances in Nucleic Acid Therapeutics
Drug Discovery Series
Editor-in-chief
David Thurston, King's College, UK
Series editors:
David Fox, Vulpine Science and Learning, UK
Ana Martinez, Centro de Investigaciones Biologicas-CSIC, Spain
David Rotella, Montclair State University, USA
Hong Shen, Roche Innovation Center Shanghai, China
Editorial advisor:
Ian Storer, AstraZeneca, UK
Titles in the Series:
1: Metabolism, Pharmacokinetics and Toxicity of Functional Groups
2: Emerging Drugs and Targets for Alzheimer's Disease; Volume 1
3: Emerging Drugs and Targets for Alzheimer's Disease; Volume 2
4: Accounts in Drug Discovery
5: New Frontiers in Chemical Biology
6: Animal Models for Neurodegenerative Disease
7: Neurodegeneration
8: G Protein-coupled Receptors
9: Pharmaceutical Process Development
10: Extracellular and Intracellular Signaling
11: New Synthetic Technologies in Medicinal Chemistry
12: New Horizons in Predictive Toxicology
13: Drug Design Strategies: Quantitative Approaches
14: Neglected Diseases and Drug Discovery
15: Biomedical Imaging
16: Pharmaceutical Salts and Cocrystals
17: Polyamine Drug Discovery
18: Proteinases as Drug Targets
19: Kinase Drug Discovery
20: Drug Design Strategies: Computational Techniques and Applications
21: Designing Multi-target Drugs
22: Nanostructured Biomaterials for Overcoming Biological Barriers
23: Physico-chemical and Computational Approaches to Drug Discovery
24: Biomarkers for Traumatic Brain Injury
25: Drug Discovery from Natural Products
26: Anti-inflammatory Drug Discovery
27: New Therapeutic Strategies for Type 2 Diabetes: Small Molecules
28: Drug Discovery for Psychiatric Disorders
29: Organic Chemistry of Drug Degradation
30: Computational Approaches to Nuclear Receptors
31: Traditional Chinese Medicine
32: Successful Strategies for the Discovery of Antiviral Drugs
33: Comprehensive Biomarker Discovery and Validation for Clinical Application
34: Emerging Drugs and Targets for Parkinson's Disease
35: Pain Therapeutics; Current and Future Treatment Paradigms
36: Biotherapeutics: Recent Developments using Chemical and Molecular Biology
37: Inhibitors of Molecular Chaperones as Therapeutic Agents
38: Orphan Drugs and Rare Diseases
39: Ion Channel Drug Discovery
40: Macrocycles in Drug Discovery
41: Human-based Systems for Translational Research
42: Venoms to Drugs: Venom as a Source for the Development of Human Therapeutics
43: Carbohydrates in Drug Design and Discovery
44: Drug Discovery for Schizophrenia
45: Cardiovascular and Metabolic Disease: Scientific Discoveries and New Therapies
46: Green Chemistry Strategies for Drug Discovery
47: Fragment-based Drug Discovery
48: Epigenetics for Drug Discovery
49: New Horizons in Predictive Drug Metabolism and Pharmacokinetics
50: Privileged Scaffolds in Medicinal Chemistry: Design, Synthesis, Evaluation
51: Nanomedicines: Design, Delivery and Detection
52: Synthetic Methods in Drug Discovery: Volume 1
53: Synthetic Methods in Drug Discovery: Volume 2
54: Drug Transporters: Role and Importance in ADME and Drug Development
55: Drug Transporters: Recent Advances and Emerging Technologies
56: Allosterism in Drug Discovery
57: Anti-aging Drugs: From Basic Research to Clinical Practice
58: Antibiotic Drug Discovery: New Targets and Molecular Entities
59: Peptide-based Drug Discovery: Challenges and New Therapeutics
60: Drug Discovery for Leishmaniasis
61: Biophysical Techniques in Drug Discovery
62: Acute Brain Impairment Through Stroke: Drug Discovery and Translational Research
63: Theranostics and Image Guided Drug Delivery
64: Pharmaceutical Formulation: The Science and Technology of Dosage Forms
65: Small-molecule Transcription Factor Inhibitors in Oncology
66: Therapies for Retinal Degeneration: Targeting Common Processes
67: Kinase Drug Discovery: Modern Approaches
68: Advances in Nucleic Acid Therapeutics
How to obtain future titles on publication: A standing order plan is available for this series. A standing order will bring delivery of each new volume immediately on publication.
For further information please contact: Book Sales Department, Royal Society of Chemistry, Thomas Graham House, Science Park, Milton Road, Cambridge, CB4 0WF, UK
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Foreword
The initial foundation for using either DNA or RNA (oligonucleotides) as therapeutic drugs was formulated by Zamecnik and Stephenson in a classic paper published several years ago (P. C. Zamecnik and M. L. Stephenson, Proc. Natl. Acad. Sci. U. S. A., 1978, 75, 280). Many of us considered that this concept would prove to be a new, refreshing approach for providing revolutionary drugs useful in the treatment of nondruggable diseases. Over time these expectations have been realized as several oligonucleotide drugs (Macugen, Fomivirsen, Mipomersen, Eteplirsen, Nusinersen, Inotersen, and Patisiran) are currently available for treating a diverse group of diseases. There are also a large number of additional oligonucleotides in various stages of drug development.
The path towards identifying drugs in the nucleic acids therapeutic arena has involved many unexpected revelations. Initially the focus was strictly on using antisense oligonucleotides. Over time, new research has opened possibilities for oligonucleotide drugs in such diverse fields as interfering RNA, microRNA, noncoding RNA, splicing modulation of RNA transcripts, targeting toxic repeats in RNA and DNA, investigating RNA and DNA aptamers and ribozymes for treating various disease states, and formulating synthetic agonists of Toll-like receptors. In this book the editors, through various chapters, provide a broad and complete perspective on the history of these fields. As each chapter unfolds, the reader discovers the logic behind why various DNA and RNA analogues were developed, how they were applied in clinical studies, and the limitations and advantages of these analogues. Also clearly presented are details on the unexpected side effects with some of these being very serious, such as the Toll receptor problem, the retention and targeting of oligonucleotides in tissues, and the variation of clinical studies with animal models.
Published by the Royal Society of Chemistry, www.rsc.org
Drug development in the nucleic acids field continues at an increasing rate. Moreover so do the challenges that are important for successfully identifying an oligonucleotide therapeutic drug. This book provides an excellent road map for navigating all that has come before and also outlines for the reader a multitude of possible directions on how to proceed with further research. The editors have assembled an excellent set of authors who are experts in the science as presented in various chapters. This would be expected as both Michael Gait and Sudhir Agrawal are among the most highly respected and experienced research scientists in this field. Thus it is not at all surprising that the science is both current and focused on important concepts. I can enthusiastically recommend this book as both a reference and also as a guide for further research in the nucleic acids therapeutics arena.
Marv Caruthers
University of Colorado
Preface
Nucleic acids therapeutics are now recognized as the third major drug discovery platform, in addition to small molecules and proteins. In the past three decades, tremendous progress has been made towards the realization of the potential of nucleic acids therapeutics for the treatment of a broad range of diseases. In addition, multiple mechanisms of actions have been elucidated. Recently, several nucleic acid drugs have been approved for clinical use.
Chemical modifications of the three components of nucleic acids – heterocyclic bases, five-membered sugars, and internucleotide linkages – as well as the nucleotide sequences themselves are the key drivers for the creation of nucleic acid drugs. Rational combinations of these have provided drug-like properties. Further advances in the chemistry of nucleic acids and additional insights into their mechanisms of action have expanded their applications to include antisense targeting of mRNA, microRNA, non-coding RNA and splicing modulation, ribozymes, RNA interference (RNAi) and short interfering RNA (siRNA), gene editing, aptamers, and the modulation of immune responses. During the past ten years, since the excellent publication of Jens Kurreck's book (Therapeutic Oligonucleotides), progress in this field has been so rapid and broad that we felt it was appropriate now to document the key developments in the field in the form of a new book.
In Chapter 1 we provide a brief history of the development of nucleotide analogues, early experience in the use of modified antisense oligonucleotides (ONs) from preclinical studies to human trials, as well as the importance of nucleotide sequence and its implications in interaction with innate immune receptors. The next three chapters provide updates on applications of antisense technology. In Chapter 2, David Corey and Zhongtian Liu discuss various mechanisms of action of antisense ONs. In Chapter 3, Eric Swayze and Punit Seth describe the medicinal chemistry of RNase H-activating antisense
Published by the Royal Society of Chemistry, www.rsc.org
ONs and in Chapter 4, Richard Geary, Brenda Baker and Brett Monia provide an update on and experience of the application of antisense ONs in clinical development.
During the development of antisense technology, it was realized that subcutaneous delivery of antisense ONs led to activation of host immune responses. Initially, this was thought to be a side effect but soon the discovery of the family of Toll-like receptors (TLRs) led to an understanding of immune activation triggered by receptor-mediated engagement. Tremendous progress has been made in translating these observations into a novel therapeutic platform. In Chapter 13, Shin-Ichiroh Saitoh and Kensuke Miyake provide a detailed background on immune receptors that are known to recognize nucleic acid sequence, patterns, and motifs. In Chapters 5 and 14, one of us (S.A.) and Ekambar Kandimalla have discussed the chemistry of novel nucleic acid compounds and how they modulate receptor-mediated immune responses, along with their therapeutic applications, including clinical proof of concept trials.
A more recent therapeutic application of antisense involves splicing modulation to affect the translation of the targeted pre-mRNA. In Chapter 6, Elena Daoutsali and Annemieke Aartsma-Rus provide an up to date survey on this subject through a variety of examples. Similarly, applications of antisense have been expanded to targeting toxic RNA repeats (Chapter 7 by Derick Wansink and colleagues), microRNA (Chapter 8 by Anna Malinowksi and Jonathan Hall) and long non-coding RNA (Chapter 9 by Claes Wahlestedt and colleagues).
In parallel, significant advances have also been made in RNAi technology for therapeutics. In Chapter 10, Anastasia Khvorova and colleagues discuss in detail the challenges of delivery of RNAi-based therapeutics and how these obstacles have been addressed. In Chapter 11, Muthiah Manoharan and Kallanthottathil Rajeev describe the clinical development of siRNA candidates targeted to liver. In Chapter 12 Jiehua Zhou and John Rossi describe the application of RNAi for treatment of HIV infection. Therapeutic application of ribozyme technology had shown early promise, but has now been found to have significant limitations. In Chapter 18, Darko Balke and Sabine Müller describe novel ribozyme constructs in the search for potential therapeutic applications.
In the past few years, we have also seen explosive growth in the development of gene editing using nucleic acids towards therapeutics. In Chapter 17, Carine Giovannangeli and colleagues provide details on this subject. Synthetic nucleic acids have been studied as aptamers to target proteins and other cellular targets and their clinical evaluation is reviewed in Chapter 15 by Paloma Giangrande and colleagues and in Chapter 16 by Gerald Zon. Through understanding the various mechanisms of actions of nucleic acids, extensive experience has been gained on their safety and pharmacokinetics, both in preclinical and in clinical use. In Chapter 20, Cathaline den Besten and Patrik Andersson discuss this topic in detail and provide their analysis.
To maintain successes in the field, significant advances have also been made in manufacturing and quality control, discussed in Chapter 19 by Yogesh Sanghvi.
We are immensely grateful to all these co-authors for their outstanding contributions that provide a detailed story of their respective subjects along with a current bibliography. We are also grateful to the editorial team at the Royal Society of Chemistry (Katie Morrey and Drew Gwilliams, Rowan Frame and Robin Driscoll) for their timely publishing and encouragement and members of the Royal Society of Chemistry staff for their assistance, as well as the many members of the Oligonucleotide Therapeutic Society, the premier professional society in the field, who have contributed to this book, which we hope will become a manual for the state of the art.
In this relatively young field of nucleic acid therapeutics, the use of their sequences to target drugs very precisely in cells and in vivo and the development of nucleic acids chemistry have been paramount and resulted in a substantial broadening of their applications. Rapidly developed and newly approved drugs are now available for the treatment of some rare diseases and other more prevalent diseases are surely following. Despite some setbacks, the list of RNA targets and approved drugs is expanding quickly. We are excited at the future prospects for this field.
Sudhir Agrawal and Michael J. Gait
3.6.4
Elena Daoutsali and Annemieke Aartsma-Rus
6.1 Introduction
6.1.1
6.1.2 Constitutive
6.2 Therapeutic Exon
6.2.1 Antisense Oligonucleotides (ASOs) and Chemical Modifications
6.2.2 Restoring
6.3 Future Perspectives, Towards Additional Approved
6.3.2
7.4.3 Antisense
7.4.4 CRISPR/Cas9, TALEN, ZFN and Other Protein Effectors
7.5 Challenges for Multisystemic Repeat Diseases
Chapter 8 Research and Development of Oligonucleotides Targeting MicroRNAs (miRNAs)
A. L. Malinowska and J. Hall
8.1 Introduction: MicroRNA Biogenesis and Functions
8.2 miRNAs as
The Development of AntimiR
8.3.2 New Insights into Mechanisms of Oligonucleotide-based miRNA Targeting
8.3.3 New Chemistries and Alternative Approaches for Targeting miRNAs
8.4 AntimiRs in Clinical Trials
8.4.1 AntimiRs Targeting miR-122 for Treatment of HCV
8.4.2 AntimiR-targeting of Other miRNAs in
Chapter 9 Oligonucleotide Targeting of Long Non-coding RNAs
I. Blokhin, O. Khorkova, J. Hsiao and C. Wahlestedt
9.1 Introduction
9.2 History of lncRNAs
9.3 Biology and Functions of lncRNA
9.3.1 lncRNAs as Regulators of Transcription
9.3.2 lncRNA as Regulators of Post-transcriptional Processing
9.3.3 lncRNA as Regulators of Translation
9.4 Classification of lncRNA
9.4.1 Functional Classification
9.4.2 Genomic Classification
9.5 Targeting of Long Non-coding RNA by Oligonucleotides 192
9.5.1 Antisense Oligonucleotides 192
9.5.2 siRNAs
9.5.3 CRISPR and Other Approaches 193
9.6 Therapeutic Applications 194
9.6.1 Neurology and Psychiatry
9.6.2 Oncology
9.6.3 Cardiology
9.6.4 Gastroenterology
9.7
Chapter 10 Conjugate-mediated Delivery of RNAi-based Therapeutics: Enhancing Pharmacokinetics–Pharmacodynamics Relationships of Medicinal Oligonucleotides 206 Bruno M. D. C. Godinho, Andrew H. Coles and Anastasia Khvorova
10.1 Introduction 206
10.2 Chemical Stabilization as a Prerequisite for Conjugate-mediated Delivery of siRNAs: Effects on Clearance, Distribution and Sustained Gene Silencing 207
10.3 Modulating Biodistribution of Therapeutic Oligonucleotides Using Conjugated Modalities: Targeted versus Broad Functional Delivery 213 10.3.1 Broad Functional Delivery of Conjugated siRNAs 213
10.3.2 Targeted Delivery of Conjugated siRNAs 216
10.4 Productive Delivery of Therapeutic Oligonucleotides: Overcoming the Endosomal Barrier 218
10.5 The Effects of the Route of Administration: Local versus Systemic Delivery 219
10.5.1 Local Delivery of Conjugated siRNAs 219
10.5.2 Systemic Delivery of Conjugated siRNAs 221
10.6 Enhancing PK Properties of Conjugated siRNAs: Reducing Clearance Kinetics and Accelerating Target Tissue Uptake 223
10.7 Conjugation Chemistry for RNAi-based Therapeutics: Future Perspectives 224
Kallanthottathil G. Rajeev and Muthiah Manoharan
The Role of Chemistry
Liver-specific Delivery of siRNA
Ionizable Lipid Nanoparticles (iLNPs)
11.3.2 Lipid-conjugated siRNA provided Proof of Concept for RNAi Therapeutics
Discovery of GalNAc Conjugates
ONPATTRO™ (Patisiran)
Inclisiran
Givosiran
Fitusiran
Zhou and J. J. Rossi
Chapter 13 Nucleic Acid Innate Immune Receptors
Shin-Ichiroh Saitoh and Kensuke Miyake
13.1 Introduction
13.2 Toll-like Receptors
13.2.1 TLR3 Recognizes dsRNA
13.2.2 TLR7 and TLR8 Recognize ssRNA and Guanosine or Uridine
13.2.3 TLR9 Recognizes CpG-DNA
13.2.4 Chaperones Regulate the Maturation of NA-sensing TLRs
13.2.5 Unc93B1 Regulates the Balance of TLR7 and TLR9 Responses
13.2.6 Proteolytic Cleavage of NA-sensing TLRs is Essential for Their Function
13.2.7 Trafficking of TLR7 and TLR9 is Essential for Type I Interferon Production in pDCs
13.3 Nucleic Acids Sensing in the Cytoplasm
13.3.1 Cytosolic DNA Sensors Recognize dsDNA
13.3.2 RIG-I and MDA5 Recognize dsRNA and Activate MAVS to Induce
Shambhavi Shubham, Li-Hsien Lin, Ofonime Udofot, Sven Krupse and Paloma H. Giangrande
16
G. Zon
Chapter 17 CRISPR-based Technologies for Genome Engineering:
Sylvain Geny, Elaheh Sadat Hosseini, Jean-Paul Concordet and Carine Giovannangeli
Sequence-specific CRISPR Nucleases and Improved Variants
Cas 9 from Streptococcus pyogenes, Orthologues and Variants
Expanding Targeted Functions with “CRISPR Fusions”
Chapter 19
1
History and Development of Nucleotide Analogues in Nucleic Acids Drugs
SudhIR AgRAWAl*a ANd MIChAel J. gAIT*b
aArnay Sciences llC, Shrewsbury, MA 01545, uSA; bMedical Research Council, l aboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0Qh, uK
Forty years ago, Zamecnik and Stephenson proposed the therapeutic use of antisense oligonucleotides on the basis of their finding that Rous sarcoma virus (RSV) replication could be inhibited by a synthetic oligonucleotide complimentary to the RSV genome.1 This concept opened up a new approach to drug discovery, namely an oligonucleotide binding sequence-specifically via Watson–Crick base-pairing to a complementary target RNA.
Since then, continuous progress has been made towards realizing the potential of this novel scientific approach and this has led recently to the approval of five antisense drugs. While the underlying concept of antisense is very simple, a rigorous understanding of the chemistry of nucleic acids had to be developed for its use in humans. In this chapter we discuss the history of this chemistry of oligonucleotides in antisense and the lessons
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ChApTeR
learned from preclinical studies and clinical trials that have guided the development in conferring drug-like properties.
In parallel to the development of antisense (see Chapters 2–4), the application of synthetic oligonucleotides as therapeutic agents has evolved into broad applications involving multiple modalities. These applications include ribozymes (see Chapter 18), small interfering RNA (siRNA, see Chapters 10, 11 and 12), microRNA (see Chapter 8), aptamers (see Chapters 15 and 16), non-coding RNA (see Chapter 9), splicing modulation (Chapter 6), targeting toxic repeats (Chapter 7), gene editing (Chapter 17), and immune modulations (see Chapters 5, 13 and 14).
The common feature of these applications is that drug candidates are composed of natural nucleosides or nucleoside analogues linked via phosphodiester or modified linkages.
1.2 The Antisense Concept
In 1976, RSV was the only purified virus for which a sufficient quantity was available for potential sequencing. Maxam and gilbert sequenced this RNA virus and noted that both ends of the linear viral genome bore the same primary sequence and were in the same polarity. It occurred to Zamecnik that the new piece of dNA synthesized by reverse transcription at the 5ʹend of this retrovirus might circularize and hybridize with the 3ʹ-end. Thus he considered the possibility of inhibiting viral replication by adding a piece of synthetic dNA to the replication system to block the circularization step by hybridizing specifically with the 3ʹ-end of the viral RNA in a competitive way.
This experiment led to startling observations, including the inhibition of new virus particles and the prevention of transformation of chick fibroblasts into sarcoma cells. In a cell-free system, translation of the Rous sarcoma viral message was also dramatically impaired. These observations were the first to show proof of the antisense concept.1,2
Not much further progress was made in the field up to 1985, primarily for three reasons. First, there was still widespread disbelief that oligonucleotides could enter eukaryotic cells. Second, there was very little dNA (or RNA) genomic sequence available for targeting by antisense, and third, efficient automated methodologies to synthesize oligonucleotides in sufficient quantities were only just beginning to become established.
1.3 Developments in Oligonucleotide Synthesis
Although the principle of solid-phase oligonucleotide synthesis was first introduced by l etsinger and Mahadevan in 1965,3 development of more efficient methods of oligodeoxynucleotide (OdN) synthesis on solid support took place from 1975 in the g ait laboratory by the phosphodiester chemistry4 and from 1979 by the phosphotriester method in the Itakura laboratory5 and the g ait laboratory.6,7 These methods were superseded by
History and Development of Nucleotide Analogues in Nucleic Acids Drugs
the outstanding phosphoramidite chemistry of Caruthers and colleagues,8 which was automated by Applied Biosystems and other companies. This transformed the ability of non-chemists to obtain OdNs for biological purposes.
1.4 Choices for Antisense Oligodeoxynucleotide
Modifications
In the mid-1980s the discovery of human immunodeficiency virus 1 (hIV-1) and the availability of its RNA sequence led Zamecnik to employ an antisense approach in attempts to inhibit hIV replication.2 Antisense OdNs with phosphodiester linkages (pO-ASO, Figure 1.1) complementary to various regions of hIV-1 mRNA were synthesized using automated synthesizers. In hIV-1-infected cells, these pO-ASOs inhibited hIV-1 replication and suppressed expression of hIV-1 related markers.9 Such experiments were carried out using primary human cells and cellular uptake of OdNs was not a limiting factor. In these studies a control pO-ASO showed minimal inhibition of hIV-1 replication, providing evidence of sequence-specific antisense activity. These studies re-established the potential application of antisense as a therapeutic approach.
1.4.1 Backbone Modifications
It was realized that the use of pO-ASOs would have limited therapeutic application, since these ASOs would be degraded rapidly in biological fluids. Soon, the focus of research shifted into identifying novel analogues of oligonucleotides that would have increased stability against nucleases and maintain the sequence-specific hybridization for use in antisense studies. The mechanisms of ASO activities (RNase h and steric blocking) are reviewed in Chapter 2. The characteristics required for a good ASO oligonucleotide are summarized in Figure 1.2.
Figure 1.1 Chemical structures of dNA and dNA analogues (A) phosphodiester oligodeoxynucleotide (pO-OdN), (B) phosphorothioate (pS-OdN), (C) Rp pS-OdN, (d) Sp pS-OdN.
Figure 1.2 Nucleic acid-based therapeutics include the use of synthetic oligonucleotides as drug candidates. These candidates have various characteristics including dNA or RNA of varying sequence composition, single- or double-stranded, formulated, conjugated or complexed with lipid carriers etc. While the intended target of these candidates is largely RNA there are innate immune receptors including TlR 3, 7, 8 and 9, RIg-1, STINg and inflammasomes. These receptors are known to recognize patterns of nucleic acids and activate appropriate immune responses. Modulation of these receptors has shown broad therapeutic applications. In design of nucleic acid-based therapeutics and their intended mechanism of action due consideration is needed to avoid overlapping mechanisms.
1.4.1.1 Phosphorothioates
The ASO therapeutics field took a major step forward in the mid 1980s through the chemical synthesis of phosphorothioate (pS) OdNs,10 based on the pioneering work of eckstein. In these analogues a simple sulfur atom replaces an oxygen atom (Figure 1.1). pS-linked OdNs are much more resistant to nuclease degradation than phosphodiesters and thus cellular activities were found to be much higher. however, mixed diastereomeric pS-OdNs, accessible readily by automated synthesis, have lower binding affinity to target RNA as compared with pO-OdN. Synthetic methodologies were optimized to synthesize milligram quantities of pS-OdNs for use in experiments as ASOs.
Studies with pS-ASO targeted to various regions of hIV-1 mRNA as well as non-complementary analogs including homo-oligomers, were conducted in hIV-1-infected cell-based assays and showed potent dose-dependent inhibition of viral replication and antiviral activity.11,12 The antiviral activity was
History
and Development of
Nucleotide Analogues in Nucleic Acids
Drugs related to the base composition of the analogues, and longer pS oligonucleotides were more effective than shorter ones. These studies also established that in primary human cells, cellular uptake of pS-ASO was efficient and that no carrier was required. Furthermore, pS-ASOs also showed very potent and durable inhibition of hIV-1 replication in chronically hIV-1-infected cells.13–15
On the basis of the promise of these results, antisense technology gained the attention of the broader scientific community and pS-ASOs became the choice as first-generation ASOs. pS-OdNs thus quickly became the primary choice for therapeutics development by newly formed biotechnology companies, such as gilead Sciences, Isis pharmaceuticals (now Ionis), hybridon (now Idera pharmaceuticals) and several others. Over the next few years, hundreds of reports appeared in the literature on the use of pS-ASOs targeting various viruses,16–18 oncogenes,19,20 kinases19,21 and other targets.22 Soon it was realized that cellular uptake in transformed cells in culture was poor and lipid-based formulations were needed for efficient uptake and antisense activity. Also it was noted that the duplex of a pS-ASO with a target RNA elicited RNase h activity,14,15 thereby allowing pS-ASOs to cleave RNA strands and thus to inhibit translation. This was more efficient than by using a steric blocking mechanism.23,24 however the efficiency of RNase h cleavage of RNA by a pS-ASO was poor as compared with a p O-ASO.14,15
The first pharmacokinetic and tissue disposition study of a systemically delivered p S-ASO occurred in mice and showed that the plasma halflife was very short and that there was a broad tissue distribution. 25 The highest concentrations of the administered p S-ASO were observed in the liver and kidneys, with the lowest concentrations in the brain. d elivered p S-ASO remained stable in tissues for days and was excreted primarily in urine in degraded form. d egradation was primarily from the 3 ʹ -end, and modifications of the 3 ʹ -end increased the stability further. 26,27 The binding of p S-ASO to serum proteins played a major role in plasma halflife and tissue disposition and was affected by the presence of secondary structures. 28,29
Studies with multiple p S-ASOs of varying sequence composition and length were conducted in animal models of viral diseases and cancer.30,31 In these studies p S-ASOs exerted very potent activity. h owever it soon become clear that in some cases the control p S-ASOs employed also had some activity, leading to questions on what a good control would be for p S-ASO and/or if a p S-ASO had off-target activity. 32 In one study a p S-ASO targeted to human papillomavirus inhibited papillomavirus-induced growth of implanted human foreskin in a mouse xenograft model. h owever, it also showed activity in a lethal mouse cytomegalovirus (CMV) model, in which the p S-ASO was not expected to have antisense activity. 33 d etailed studies in immune-compromised mice led to an understanding that the antiviral activity of p S-ASO was largely due to immune activation of the host.
The impact of immune-stimulatory properties of pS-ASOs also became evident during the non-clinical safety evaluations of multiple candidates in support of investigational supporting studies. In mice and rats, repeated systemic administration of pS-ASO candidates caused general inflammation, splenomegaly, thrombocytopenia, elevation of transaminases and histological changes in multiple organs.34,35 In addition, in primates, bolus administration of the first pS-ASO (geM91) led to severe hemodynamic changes, which subsequently were found to be due to activation of the alternative complement pathway.36 Complement activation thus became recognized as a second off-target effect of pS-ASOs. The alternative pathway complement activation cascade was attributed to the poly-anionic nature of pS-ASOs and largely found to be a plasma concentration threshold effect. As such, it could be mitigated by subcutaneous administration or by slow intravenous infusion to keep a low plasma concentration. Based on this observation, the uS Food and drug Administration (FdA) implemented guidelines on dosing of pS-ASOs and recommended use of non-human primates as the non-rodent species for non-clinical safety studies.37 Meanwhile, early development programs with pS-ASOs continued to proliferate.
Over 25 pS-ASO drug candidates targeted to viral RNA, oncogenes and cellular targets had advanced to human trials.38 Routes of administration included intravitreal, intravenous infusion or subcutaneous.39–41 In humans the plasma half-life and excretion of pS-ASOs was similar to that which was observed in pre-clinical models.42,43 Clinical development of most of the pS-ASO drug candidates were discontinued, either due to lack of activity or a poor therapeutic index.44 experience with a specific pS-ASO drug candidate, geM91, in humans was very informative. Subcutaneous administration of geM91 caused flu-like symptoms, swelling of the draining lymph nodes, prolongation of activated partial thromboplastin time (apTT) and more importantly, rather than suppressing hIV-1, it increased hIV-1 RNA levels in blood.45 Importantly, administration of the same dose by the intravenous route had minimal effects on these parameters. This was puzzling at that time, but much later it became clear that pS-ASO containing the unmethylated Cpg motif were activating the immune responses by binding to Toll-like receptor 9 (TlR9), an innate immune receptor present in immune cells that recognizes dNA containing Cpg dinucleotide motifs (see Chapter 14). Treatment with many other pS-ASO drug candidates also had shown flu-like symptoms and injection-site reactions. It is important to note that most of the pS-ASO drug candidates that had been advanced to human studies contained a Cpg motif.46 For example, the pS-ASO Fomivirsen, targeted to CMV and delivered intravitreally, had been approved but is no longer marketed. The mechanism of action of Fomivirsen has been questioned.
Collectively, the development of pS-ASOs from preclinical to clinical studies has provided very important insights into the properties of pS-ASOs.47–49 These could be classified as a class effect, including an affinity to target RNA, stability towards nucleases, serum protein binding and poly-anionic-related
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their own strength; that women are strong by sentiment; that the same mental height which their husbands attain by toil, they attain by sympathy with their husbands. Man is the will, and Woman the sentiment. In this ship of humanity, Will is the rudder, and Sentiment the sail: when Woman affects to steer, the rudder is only a masked sail. When women engage in any art or trade, it is usually as a resource, not as a primary object. The life of the affections is primary to them, so that there is usually no employment or career which they will not with their own applause and that of society quit for a suitable marriage. And they give entirely to their affections, set their whole fortune on the die, lose themselves eagerly in the glory of their husbands and children. Man stands astonished at a magnanimity he cannot pretend to. Mrs. Lucy Hutchinson, one of the heroines of the English Commonwealth, who wrote the life of her husband, the Governor of Nottingham, says, “If he esteemed her at a higher rate than she in herself could have deserved, he was the author of that virtue he doted on, while she only reflected his own glories upon him. All that she was, was him, while he was hers, and all that she is now, at best, but his pale shade.” As for Plato’s opinion, it is true that, up to recent times, in no art or science, nor in painting, poetry or music, have they produced a masterpiece. Till the new education and larger opportunities of very modern times, this position, with the fewest possible exceptions, has always been true. Sappho, to be sure, in the Olympic Games, gained the crown over Pindar. But, in general, no mastery in either of the fine arts—which should, one would say, be the arts of women—has yet been obtained by them, equal to the mastery of men in the same. The part they play in education, in the care of the young and the tuition of older children, is their organic office in the world. So much sympathy as they have makes them inestimable as the mediators between those who have knowledge and those who want it: besides, their fine organization, their taste and love of details, makes the knowledge they give better in their hands.
But there is an art which is better than painting, poetry, music, or architecture,—better than botany, geology, or any science; namely, Conversation. Wise, cultivated, genial conversation is the last flower of civilization and the best result which life has to offer us,—a cup for
gods, which has no repentance. Conversation is our account of ourselves. All we have, all we can, all we know, is brought into play, and as the reproduction, in finer form, of all our havings.
Women are, by this and their social influence, the civilizers of mankind. What is civilization? I answer, the power of good women. It was Burns’s remark when he first came to Edinburgh that between the men of rustic life and the polite world he observed little difference; that in the former, though unpolished by fashion and unenlightened by science, he had found much observation and much intelligence; but a refined and accomplished woman was a being almost new to him, and of which he had formed a very inadequate idea. “I like women,” said a clear-headed man of the world; “they are so finished.” They finish society, manners, language. Form and ceremony are their realm. They embellish trifles. All these ceremonies that hedge our life around are not to be despised, and when we have become habituated to them, cannot be dispensed with. No woman can despise them with impunity. Their genius delights in ceremonies, in forms, in decorating life with manners, with properties, order and grace. They are, in their nature, more relative; the circumstance must always be fit; out of place they lose half their weight, out of place they are disfranchised. Position, Wren said, is essential to the perfecting of beauty;—a fine building is lost in a dark lane; a statue should stand in the air; much more true is it of woman.
We commonly say that easy circumstances seem somehow necessary to the finish of the female character: but then it is to be remembered that they create these with all their might. They are always making that civilization which they require; that state of art, of decoration, that ornamental life in which they best appear
The spiritual force of man is as much shown in taste, in his fancy and imagination,—attaching deep meanings to things and to arbitrary inventions of no real value,—as in his perception of truth. He is as much raised above the beast by this creative faculty as by any other. The horse and ox use no delays; they run to the river when thirsty, to the corn when hungry, and say no thanks, but fight down whatever opposes their appetite. But man invents and adorns all he does with delays and degrees, paints it all over with forms, to
please himself better; he invented majesty and the etiquette of courts and drawing-rooms; architecture, curtains, dress, all luxuries and adornments, and the elegance of privacy, to increase the joys of society. He invented marriage; and surrounded by religion, by comeliness, by all manner of dignities and renunciations, the union of the sexes.
And how should we better measure the gulf between the best intercourse of men in old Athens, in London, or in our American capitals,—between this and the hedgehog existence of diggers of worms, and the eaters of clay and offal,—than by signalizing just this department of taste or comeliness? Herein woman is the prime genius and ordainer. There is no grace that is taught by the dancingmaster, no style adopted into the etiquette of courts, but was first the whim and the mere action of some brilliant woman, who charmed beholders by this new expression, and made it remembered and copied. And I think they should magnify their ritual of manners.[202] Society, conversation, decorum, flowers, dances, colors, forms, are their homes and attendants. They should be found in fit surroundings —with fair approaches, with agreeable architecture, and with all advantages which the means of man collect:
“The far-fetched diamond finds its home Flashing and smouldering in her hair. For her the seas their pearls reveal, Art and strange lands her pomp supply With purple, chrome and cochineal, Ochre and lapis lazuli. The worm its golden woof presents. Whatever runs, flies, dives or delves All doff for her their ornaments, Which suit her better than themselves.”[203]
There is no gift of Nature without some drawback. So, to women, this exquisite structure could not exist without its own penalty. More vulnerable, more infirm, more mortal than men, they could not be such excellent artists in this element of fancy if they did not lend and give themselves to it. They are poets who believe their own poetry.
They emit from their pores a colored atmosphere, one would say, wave upon wave of rosy light, in which they walk evermore, and see all objects through this warm-tinted mist that envelops them.
But the starry crown of woman is in the power of her affection and sentiment, and the infinite enlargements to which they lead. Beautiful is the passion of love, painter and adorner of youth and early life: but who suspects, in its blushes and tremors, what tragedies, heroisms and immortalities are beyond it? The passion, with all its grace and poetry, is profane to that which follows it. All these affections are only introductory to that which is beyond, and to that which is sublime.
We men have no right to say it, but the omnipotence of Eve is in humility. The instincts of mankind have drawn the Virgin Mother—
“Created beings all in lowliness Surpassing, as in height above them all.”[204]
This is the Divine Person whom Dante and Milton saw in vision. This is the victory of Griselda, her supreme humility. And it is when love has reached this height that all our pretty rhetoric begins to have meaning. When we see that, it adds to the soul a new soul, it is honey in the mouth, music in the ear and balsam in the heart.
“Far have I clambered in my mind, But nought so great as Love I find. What is thy tent, where dost thou dwell?
‘My mansion is humility, Heaven’s vastest capability.’
The further it doth downward tend, The higher up it doth ascend.”[205]
The first thing men think of, when they love, is to exhibit their usefulness and advantages to the object of their affection. Women make light of these, asking only love. They wish it to be an exchange of nobleness.
There is much in their nature, much in their social position which gives them a certain power of divination. And women know, at first
sight, the characters of those with whom they converse. There is much that tends to give them a religious height which men do not attain. Their sequestration from affairs and from the injury to the moral sense which affairs often inflict, aids this. And in every remarkable religious development in the world, women have taken a leading part. It is very curious that in the East, where Woman occupies, nationally, a lower sphere, where the laws resist the education and emancipation of women,—in the Mohammedan faith, Woman yet occupies the same leading position, as a prophetess, that she has among the ancient Greeks, or among the Hebrews, or among the Saxons. This power, this religious character, is everywhere to be remarked in them.[206]
The action of society is progressive. In barbarous society the position of women is always low—in the Eastern nations lower than in the West. “When a daughter is born,” says the Shiking, the old Sacred Book of China, “she sleeps on the ground, she is clothed with a wrapper, she plays with a tile; she is incapable of evil or of good.” And something like that position, in all low society, is the position of woman; because, as before remarked, she is herself its civilizer. With the advancements of society, the position and influence of woman bring her strength or her faults into light. In modern times, three or four conspicuous instrumentalities may be marked. After the deification of Woman in the Catholic Church, in the sixteenth or seventeenth century,—when her religious nature gave her, of course, new importance,—the Quakers have the honor of having first established, in their discipline, the equality in the sexes. It is even more perfect in the later sect of the Shakers, where no business is broached or counselled without the intervention of one elder and one elderess.
A second epoch for Woman was in France,—entirely civil; the change of sentiment from a rude to a polite character, in the age of Louis XIV.,—commonly dated from the building of the Hôtel de Rambouillet.[207] I think another important step was made by the doctrine of Swedenborg, a sublime genius who gave a scientific exposition of the part played severally by man and woman in the world, and showed the difference of sex to run through nature and
through thought. Of all Christian sects this is at this moment the most vital and aggressive.
Another step was the effect of the action of the age in the antagonism to Slavery. It was easy to enlist Woman in this; it was impossible not to enlist her. But that Cause turned out to be a great scholar. He was a terrible metaphysician. He was a jurist, a poet, a divine. Was never a University of Oxford or Göttingen that made such students. It took a man from the plough and made him acute, eloquent, and wise, to the silencing of the doctors. There was nothing it did not pry into, no right it did not explore, no wrong it did not expose. And it has, among its other effects, given Woman a feeling of public duty and an added self-respect.
One truth leads in another by the hand; one right is an accession of strength to take more. And the times are marked by the new attitude of Woman; urging, by argument and by association, her rights of all kinds,—in short, to one half of the world;—as the right to education, to avenues of employment, to equal rights of property, to equal rights in marriage, to the exercise of the professions and of suffrage.
Of course, this conspicuousness had its inconveniences. But it is cheap wit that has been spent on this subject; from Aristophanes, in whose comedies I confess my dulness to find good joke, to Rabelais, in whom it is monstrous exaggeration of temperament, and not borne out by anything in nature,—down to English Comedy, and, in our day, to Tennyson,[208] and the American newspapers. In all, the body of the joke is one, namely, to charge women with temperament; to describe them as victims of temperament; and is identical with Mahomet’s opinion that women have not a sufficient moral or intellectual force to control the perturbations of their physical structure. These were all drawings of morbid anatomy, and such satire as might be written on the tenants of a hospital or on an asylum for idiots. Of course it would be easy for women to retaliate in kind, by painting men from the dogs and gorillas that have worn our shape. That they have not, is an eulogy on their taste and selfrespect. The good easy world took the joke which it liked. There is always the want of thought; there is always credulity. There are
plenty of people who believe women to be incapable of anything but to cook, incapable of interest in affairs. There are plenty of people who believe that the world is governed by men of dark complexions, that affairs are only directed by such, and do not see the use of contemplative men, or how ignoble would be the world that wanted them. And so without the affection of women.
But for the general charge: no doubt it is well founded. They are victims of the finer temperament. They have tears, and gayeties, and faintings, and glooms and devotion to trifles. Nature’s end, of maternity for twenty years, was of so supreme importance that it was to be secured at all events, even to the sacrifice of the highest beauty. They are more personal. Men taunt them that, whatever they do, say, read or write, they are thinking of themselves and their set. Men are not to the same degree temperamented, for there are multitudes of men who live to objects quite out of them, as to politics, to trade, to letters or an art, unhindered by any influence of constitution.
The answer that lies, silent or spoken, in the minds of wellmeaning persons, to the new claims, is this: that though their mathematical justice is not to be denied, yet the best women do not wish these things; they are asked for by people who intellectually seek them, but who have not the support or sympathy of the truest women; and that, if the laws and customs were modified in the manner proposed, it would embarrass and pain gentle and lovely persons with duties which they would find irksome and distasteful. Very likely. Providence is always surprising us with new and unlikely instruments. But perhaps it is because these people have been deprived of education, fine companions, opportunities, such as they wished,—because they feel the same rudeness and disadvantage which offends you,—that they have been stung to say, ‘It is too late for us to be polished and fashioned into beauty, but, at least, we will see that the whole race of women shall not suffer as we have suffered.’
They have an unquestionable right to their own property. And if a woman demand votes, offices and political equality with men, as among the Shakers an Elder and Elderess are of equal power,—and
among the Quakers,—it must not be refused. It is very cheap wit that finds it so droll that a woman should vote. Educate and refine society to the highest point,—bring together a cultivated society of both sexes, in a drawing-room, and consult and decide by voices on a question of taste or on a question of right, and is there any absurdity or any practical difficulty in obtaining their authentic opinions? If not, then there need be none in a hundred companies, if you educate them and accustom them to judge. And, for the effect of it, I can say, for one, that all my points would sooner be carried in the State if women voted. On the questions that are important,—whether the government shall be in one person, or whether representative, or whether democratic; whether men shall be holden in bondage, or shall be roasted alive and eaten, as in Typee, or shall be hunted with bloodhounds, as in this country; whether men shall be hanged for stealing, or hanged at all; whether the unlimited sale of cheap liquors shall be allowed;—they would give, I suppose, as intelligent a vote as the voters of Boston or New York.
We may ask, to be sure,—Why need you vote? If new power is here, of a character which solves old tough questions, which puts me and all the rest in the wrong, tries and condemns our religion, customs, laws, and opens new careers to our young receptive men and women, you can well leave voting to the old dead people. Those whom you teach, and those whom you half teach, will fast enough make themselves considered and strong with their new insight, and votes will follow from all the dull.
The objection to their voting is the same as is urged, in the lobbies of legislatures, against clergymen who take an active part in politics; —that if they are good clergymen they are unacquainted with the expediencies of politics, and if they become good politicians they are worse clergymen. So of women, that they cannot enter this arena without being contaminated and unsexed.
Here are two or three objections: first, a want of practical wisdom; second, a too purely ideal view; and, third, danger of contamination. For their want of intimate knowledge of affairs, I do not think this ought to disqualify them from voting at any town-meeting which I ever attended. I could heartily wish the objection were sound. But if
any man will take the trouble to see how our people vote,—how many gentlemen are willing to take on themselves the trouble of thinking and determining for you, and, standing at the door of the polls, give every innocent citizen his ticket as he comes in, informing him that this is the vote of his party; and how the innocent citizen, without further demur, goes and drops it in the ballot-box,—I cannot but think he will agree that most women might vote as wisely.
For the other point, of their not knowing the world, and aiming at abstract right without allowance for circumstances,—that is not a disqualification, but a qualification. Human society is made up of partialities. Each citizen has an interest and a view of his own, which, if followed out to the extreme, would leave no room for any other citizen. One man is timid and another rash; one would change nothing, and the other is pleased with nothing; one wishes schools, another armies, one gunboats, another public gardens. Bring all these biases together and something is done in favor of them all.
Every one is a half vote, but the next elector behind him brings the other or corresponding half in his hand: a reasonable result is had. Now there is no lack, I am sure, of the expediency, or of the interests of trade or of imperative class interests being neglected. There is no lack of votes representing the physical wants; and if in your city the uneducated emigrant vote numbers thousands, representing a brutal ignorance and mere animal wants, it is to be corrected by an educated and religious vote, representing the wants and desires of honest and refined persons. If the wants, the passions, the vices, are allowed a full vote through the hands of a half-brutal intemperate population, I think it but fair that the virtues, the aspirations should be allowed a full vote, as an offset, through the purest part of the people.
As for the unsexing and contamination,—that only accuses our existing politics, shows how barbarous we are,—that our policies are so crooked, made up of things not to be spoken, to be understood only by wink and nudge; this man to be coaxed, that man to be bought, and that other to be duped. It is easy to see that there is contamination enough, but it rots the men now, and fills the air with stench. Come out of that: it is like a dance-cellar. The fairest names
in this country in literature, in law, have gone into Congress and come out dishonored. And when I read the list of men of intellect, of refined pursuits, giants in law, or eminent scholars, or of social distinction, leading men of wealth and enterprise in the commercial community, and see what they have voted for and suffered to be voted for, I think no community was ever so politely and elegantly betrayed.
I do not think it yet appears that women wish this equal share in public affairs. But it is they and not we that are to determine it. Let the laws be purged of every barbarous remainder, every barbarous impediment to women. Let the public donations for education be equally shared by them, let them enter a school as freely as a church, let them have and hold and give their property as men do theirs;—and in a few years it will easily appear whether they wish a voice in making the laws that are to govern them. If you do refuse them a vote, you will also refuse to tax them,—according to our Teutonic principle, No representation, no tax.
All events of history are to be regarded as growths and offshoots of the expanding mind of the race, and this appearance of new opinions, their currency and force in many minds, is itself the wonderful fact. For whatever is popular is important, shows the spontaneous sense of the hour. The aspiration of this century will be the code of the next. It holds of high and distant causes, of the same influences that make the sun and moon. When new opinions appear, they will be entertained and respected, by every fair mind, according to their reasonableness, and not according to their convenience, or their fitness to shock our customs. But let us deal with them greatly; let them make their way by the upper road, and not by the way of manufacturing public opinion, which lapses continually into expediency, and makes charlatans. All that is spontaneous is irresistible, and forever it is individual force that interests. I need not repeat to you—your own solitude will suggest it—that a masculine woman is not strong, but a lady is. The loneliest thought, the purest prayer, is rushing to be the history of a thousand years.
Let us have the true woman, the adorner, the hospitable, the religious heart, and no lawyer need be called in to write stipulations,
the cunning clauses of provision, the strong investitures;—for woman moulds the lawgiver and writes the law. But I ought to say, I think it impossible to separate the interests and education of the sexes. Improve and refine the men, and you do the same by the women, whether you will or no. Every woman being the wife or the daughter of a man,—wife, daughter, sister, mother, of a man, she can never be very far from his ear, never not of his counsel, if she has really something to urge that is good in itself and agreeable to nature. Slavery it is that makes slavery; freedom, freedom. The slavery of women happened when the men were slaves of kings. The melioration of manners brought their melioration of course. It could not be otherwise, and hence the new desire of better laws. For there are always a certain number of passionately loving fathers, brothers, husbands and sons who put their might into the endeavor to make a daughter, a wife, or a mother happy in the way that suits best. Woman should find in man her guardian. Silently she looks for that, and when she finds that he is not, as she instantly does, she betakes her to her own defences, and does the best she can. But when he is her guardian, fulfilled with all nobleness, knows and accepts his duties as her brother, all goes well for both.
The new movement is only a tide shared by the spirits of man and woman; and you may proceed in the faith that whatever the woman’s heart is prompted to desire, the man’s mind is simultaneously prompted to accomplish.[209]
XXI ADDRESS
TO THE INHABITANTS OF CONCORD AT THE CONSECRATION OF SLEEPY HOLLOW
SEPTEMBER 29, 1855
SLEEPY HOLLOW
“No abbey’s gloom, nor dark cathedral stoops, No winding torches paint the midnight air; Here the green pines delight, the aspen droops Along the modest pathways, and those fair Pale asters of the season spread their plumes Around this field, fit garden for our tombs.
And shalt thou pause to hear some funeral-bell Slow stealing o’er the heart in this calm place, Not with a throb of pain, a feverish knell, But in its kind and supplicating grace, It says, Go, pilgrim, on thy march, be more Friend to the friendless than thou wast before;
Learn from the loved one’s rest serenity; To-morrow that soft bell for thee shall sound, And thou repose beneath the whispering tree, One tribute more to this submissive ground;— Prison thy soul from malice, bar out pride, Nor these pale flowers nor this still field deride:
Rather to those ascents of being turn
Where a ne’er-setting sun illumes the year Eternal, and the incessant watch-fires burn Of unspent holiness and goodness clear,— Forget man’s littleness, deserve the best, God’s mercy in thy thought and life contest.”
W E C .
ADDRESS
TO THE INHABITANTS OF CONCORD AT THE CONSECRATION OF SLEEPY
HOLLOW SEPTEMBER 29, 1855
C : The committee to whom was confided the charge of carrying out the wishes of the town in opening the cemetery, having proceeded so far as to enclose the ground, and cut the necessary roads, and having laid off as many lots as are likely to be wanted at present, have thought it fit to call the inhabitants together, to show you the ground, now that the new avenues make its advantages appear; and to put it at your disposition.
They have thought that the taking possession of this field ought to be marked by a public meeting and religious rites: and they have requested me to say a few words which the serious and tender occasion inspires.
And this concourse of friendly company assures me that they have rightly interpreted your wishes. [Here followed, in the address, about three pages of matter which Mr. Emerson used later in his essay on Immortality, which may be found in the volume Letters and Social Aims, beginning on page 324, “The credence of men,” etc., and ending on pages 326-27 with the sentence, “Meantime the true disciples saw, through the letters, the doctrine of eternity which dissolved the poor corpse and nature also, and gave grandeur to the passing hour.”]
In these times we see the defects of our old theology; its inferiority to our habit of thoughts. Men go up and down; Science is popularized; the irresistible democracy—shall I call it?—of chemistry, of vegetation, which recomposes for new life every decomposing particle,—the race never dying, the individual never spared,—have impressed on the mind of the age the futility of these old arts of preserving. We give our earth to earth. We will not jealously guard a few atoms under immense marbles, selfishly and impossibly sequestering it from the vast circulations of Nature, but, at the same time, fully admitting the divine hope and love which belong to our nature, wishing to make one spot tender to our children, who shall come hither in the next century to read the dates of these lives.
Our people accepting this lesson from science, yet touched by the tenderness which Christianity breathes, have found a mean in the consecration of gardens. A simultaneous movement has, in a hundred cities and towns in this country, selected some convenient piece of undulating ground with pleasant woods and waters; every family chooses its own clump of trees; and we lay the corpse in these leafy colonnades.
A grove of trees,—what benefit or ornament is so fair and great? they make the landscape; they keep the earth habitable; their roots run down, like cattle, to the water-courses; their heads expand to feed the atmosphere. The life of a tree is a hundred and a thousand years; its decays ornamental; its repairs self-made: they grow when we sleep, they grew when we were unborn. Man is a moth among these longevities. He plants for the next millennium. Shadows haunt them; all that ever lived about them cling to them. You can almost see behind these pines the Indian with bow and arrow lurking yet exploring the traces of the old trail.
Modern taste has shown that there is no ornament, no architecture alone, so sumptuous as well disposed woods and waters, where art has been employed only to remove superfluities, and bring out the natural advantages. In cultivated grounds one sees the picturesque and opulent effect of the familiar shrubs, barberry, lilac, privet and thorns, when they are disposed in masses, and in large spaces. What work of man will compare with the plantation of a park? It
dignifies life. It is a seat for friendship, counsel, taste and religion. I do not wonder that they are the chosen badge and point of pride of European nobility. But how much more are they needed by us, anxious, overdriven Americans, to stanch and appease that fury of temperament which our climate bestows!
This tract fortunately lies adjoining to the Agricultural Society’s ground, to the New Burial Ground, to the Court House and the Town House, making together a large block of public ground, permanent property of the town and county,—all the ornaments of either adding so much value to all.
I suppose all of us will readily admit the value of parks and cultivated grounds to the pleasure and education of the people, but I have heard it said here that we would gladly spend for a park for the living, but not for a cemetery; a garden for the living, a home of thought and friendship. Certainly the living need it more than the dead; indeed, to speak precisely, it is given to the dead for the reaction of benefit on the living. But if the direct regard to the living be thought expedient, that is also in your power. This ground is happily so divided by Nature as to admit of this relation between the Past and the Present. In the valley where we stand will be the Monuments. On the other side of the ridge, towards the town, a portion of the land is in full view of the cheer of the village and is out of sight of the Monuments; it admits of being reserved for secular purposes; for games,—not such as the Greeks honored the dead with, but for games of education; the distribution of school prizes; the meeting of teachers; patriotic eloquence, the utterance of the principles of national liberty to private, social, literary or religious fraternities. Here we may establish that most agreeable of all museums, and agreeable to the temper of our times,—an Arboretum,—wherein may be planted, by the taste of every citizen, one tree, with its name recorded in a book; every tree that is native to Massachusetts, or will grow in it; so that every child may be shown growing, side by side, the eleven oaks of Massachusetts; and the twenty willows; the beech, which we have allowed to die out of the eastern counties; and here the vast firs of California and Oregon.
This spot for twenty years has borne the name of Sleepy Hollow Its seclusion from the village in its immediate neighborhood had made it to all the inhabitants an easy retreat on a Sabbath day, or a summer twilight, and it was inevitably chosen by them when the design of a new cemetery was broached, if it did not suggest the design, as the fit place for their final repose. In all the multitudes of woodlands and hillsides, which within a few years have been laid out with a similar design, I have not known one so fitly named. Sleepy Hollow. In this quiet valley, as in the palm of Nature’s hand, we shall sleep well when we have finished our day What is the Earth itself but a surface scooped into nooks and caves of slumber—according to the Eastern fable, a bridge full of holes, into one or other of which all the passengers sink to silence? Nay, when I think of the mystery of life, its round of illusions, our ignorance of its beginning or its end, the speed of the changes of that glittering dream we call existence, —I think sometimes that the vault of the sky arching there upward, under which our busy being is whirled, is only a Sleepy Hollow, with path of Suns, instead of foot-paths; and Milky Ways, for truck-roads.
The ground has the peaceful character that belongs to this town;— no lofty crags, no glittering cataracts;—but I hold that every part of Nature is handsome when not deformed by bad Art. Bleak sea-rocks and sea-downs and blasted heaths have their own beauty; and though we make much ado in our praises of Italy or Andes, Nature makes not so much difference. The morning, the moonlight, the spring day, are magical painters, and can glorify a meadow or a rock.
But we must look forward also, and make ourselves a thousand years old; and when these acorns, that are falling at our feet, are oaks overshadowing our children in a remote century, this mute green bank will be full of history: the good, the wise and great will have left their names and virtues on the trees; heroes, poets, beauties, sanctities, benefactors, will have made the air timeable and articulate.
And hither shall repair, to this modest spot of God’s earth, every sweet and friendly influence; the beautiful night and beautiful day will come in turn to sit upon the grass. Our use will not displace the old tenants. The well-beloved birds will not sing one song the less, the
high-holding woodpecker, the meadow-lark, the oriole, robin, purple finch, bluebird, thrush and red-eyed warbler, the heron, the bittern will find out the hospitality and protection from the gun of this asylum, and will seek the waters of the meadow; and in the grass, and by the pond, the locust, the cricket and the hyla, shall shrilly play.
We shall bring hither the body of the dead, but how shall we catch the escaped soul? Here will burn for us, as the oath of God, the sublime belief. I have heard that death takes us away from ill things, not from good. I have heard that when we pronounce the name of man, we pronounce the belief of immortality. All great natures delight in stability; all great men find eternity affirmed in the promise of their faculties. Why is the fable of the Wandering Jew agreeable to men, but because they want more time and land to execute their thoughts in? Life is not long enough for art, nor long enough for friendship. The evidence from intellect is as valid as the evidence from love. The being that can share a thought and feeling so sublime as confidence in truth is no mushroom. Our dissatisfaction with any other solution is the blazing evidence of immortality.
XXII
ROBERT BURNS
SPEECH DELIVERED AT THE CELEBRATION OF THE BURNS CENTENARY, BOSTON JANUARY
25, 1859
“His was the music to whose tone The common pulse of man keeps time In cot or castle’s mirth or moan, In cold or sunny clime.
Praise to the bard! his words are driven, Like flower-seeds by the far winds sown, Where’er, beneath the sky of heaven, The birds of fame have flown.”
H .
ROBERT BURNS
MR. PRESIDENT, G : I do not know by what untoward accident it has chanced, and I forbear to inquire, that, in this accomplished circle, it should fall to me, the worst Scotsman of all, to receive your commands, and at the latest hour too, to respond to the sentiment just offered, and which indeed makes the occasion. But I am told there is no appeal, and I must trust to the inspirations of the theme to make a fitness which does not otherwise exist. Yet, Sir,
I heartily feel the singular claims of the occasion. At the first announcement, from I know not whence, that the 25th of January was the hundredth anniversary of the birth of Robert Burns, a sudden consent warmed the great English race, in all its kingdoms, colonies and states, all over the world, to keep the festival. We are here to hold our parliament with love and poesy, as men were wont to do in the Middle Ages. Those famous parliaments might or might not have had more stateliness and better singers than we,—though that is yet to be known,—but they could not have better reason. I can only explain this singular unanimity in a race which rarely acts together, but rather after their watch-word, Each for himself,—by the fact that Robert Burns, the poet of the middle class, represents in the mind of men to-day that great uprising of the middle class against the armed and privileged minorities, that uprising which worked politically in the American and French Revolutions, and which, not in governments so much as in education and social order, has changed the face of the world.
In order for this destiny, his birth, breeding and fortunes were low. His organic sentiment was absolute independence, and resting as it should on a life of labor. No man existed who could look down on him. They that looked into his eyes saw that they might look down the sky as easily.[210] His muse and teaching was common sense, joyful, aggressive, irresistible. Not Latimer, nor Luther struck more telling blows against false theology than did this brave singer. The Confession of Augsburg, the Declaration of Independence, the French Rights of Man, and the Marseillaise, are not more weighty documents in the history of freedom than the songs of Burns. His satire has lost none of its edge. His musical arrows yet sing through the air. He is so substantially a reformer that I find his grand plain sense in close chain with the greatest masters,—Rabelais, Shakspeare in comedy, Cervantes, Butler, and Burns. If I should add another name, I find it only in a living countryman of Burns.[211]
He is an exceptional genius. The people who care nothing for literature and poetry care for Burns. It was indifferent—they thought who saw him—whether he wrote verse or not: he could have done anything else as well. Yet how true a poet is he! And the poet, too, of
poor men, of gray hodden and the guernsey coat and the blouse. He has given voice to all the experiences of common life; he has endeared the farmhouse and cottage, patches and poverty, beans and barley; ale, the poor man’s wine; hardship; the fear of debt; the dear society of weans and wife, of brothers and sisters, proud of each other, knowing so few and finding amends for want and obscurity in books and thoughts.[212] What a love of Nature, and, shall I say it? of middle-class Nature. Not like Goethe, in the stars, or like Byron, in the ocean, or Moore, in the luxurious East, but in the homely landscape which the poor see around them,—bleak leagues of pasture and stubble, ice and sleet and rain and snow-choked brooks; birds, hares, field-mice, thistles and heather, which he daily knew. How many “Bonny Doons” and “John Anderson my jo’s” and “Auld lang synes” all around the earth have his verses been applied to! And his love-songs still woo and melt the youths and maids; the farm-work, the country holiday, the fishing-cobble are still his debtors to-day.
And as he was thus the poet of the poor, anxious, cheerful, working humanity, so had he the language of low life. He grew up in a rural district, speaking a patois unintelligible to all but natives, and he has made the Lowland Scotch a Doric dialect of fame. It is the only example in history of a language made classic by the genius of a single man. But more than this. He had that secret of genius to draw from the bottom of society the strength of its speech, and astonish the ears of the polite with these artless words, better than art, and filtered of all offence through his beauty. It seemed odious to Luther that the devil should have all the best tunes; he would bring them into the churches; and Burns knew how to take from fairs and gypsies, blacksmiths and drovers, the speech of the market and street, and clothe it with melody. But I am detaining you too long. The memory of Burns,—I am afraid heaven and earth have taken too good care of it to leave us anything to say. The west winds are murmuring it. Open the windows behind you, and hearken for the incoming tide, what the waves say of it. The doves perching always on the eaves of the Stone Chapel opposite, may know something about it. Every name in broad Scotland keeps his fame bright. The
