LIPID BASED NANOFORMULATION APPROACH TO TARGET BRAIN FOR THE MANAGEMENT OF TUBERCULOSIS THROUGH INTRANASAL DELIVERY: FORMULATION DEVELOPMENT AND EVALUATION Dr. R. Suresh Kumar Department of Pharmaceutics, JSS College of pharmacy Ooty, Tamilnadu (JSS Academy of Higher Education And Research Mysuru) Abstract BACKGROUND: Tuberculosis is caused by Mycobacterium tuberculosis. It usually affects the lungs but can enter the oxygenated regions and as brain is one among them it enters the brain and leads to tuberculous meningitis. There are any drugs available for the treatment but the main problem is that the drugs cannot enter the brain by crossing the BBB METHODOLOGY: Spontaneous emulsification method was used in preparing the nanoemulsion. The compatibility studies i.e., DSCnd FTIR were carried out for isoniazid, rifampicin, resveratrol and physical mixture Then the pure drugs and the formulations were given for cell-line studies to know the cytotoxicity. The formulations were given for histopathological studies. Then these formulations were administered to rats intranasally and the brain of the rats was collected. Then the brain tissue was minced and was given for LCMS analysis to know the amount of drug reaching the brain. RESULT: The DSC and FTIR graphs state that the drugs were compatible with each other. The formulations showed low nasal irritancy and no malignancy. From the LCMS analysis it was found that 82.6% of Isoniazid= Resveratrol nanoemulsion and 78.48% of Rifampicin + Resveratrol nanoemulsion has reached the targeted area the brain. CONCLUSION: From the studies it can be concluded that when the formulations were administered intranasally maximum amount of the drugs had reached the targeted area the brain. So, when the drugs are given intranasally there could be decrease in the dose of the drug, increased bioavailability at the site of action and increased therapeutic efficacy.
Introduction: Tuberculosis is caused by the bacteria Mycobacterium Tuberculosis. It usually affects the lungs which is termed as pulmonary tuberculosis. This bacteria has the ability to enter the oxygenated regions of the body. As, brain is one among the oxygenated regions the bacteria enter the brain leading to tuberculous meningitis. There are many drugs available for the treatment of tuberculous meningitis but very less amount of drug reaches the targeted area the brain. Most of the drugs wont reach the targeted area due to the presence of the Blood Brain Barrier. The time period of the treatment is also very high. So, due to this reason we are formulating the drugs into a lipid based nanoemulsion . The prepared formulation was administered intranasally. It was administered through intranasal route because of the presence of the olfactory nerves in the brain which end in the olfactory bulb in the nasal cavity.
RESULTS
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15 10
12 5
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OLEIC ACID
ISO PROPYL MYRISTATE
OLIVE OIL
SOYA BEAN OIL
LABRAFAC PG
CORN OIL
CAPMUL MCM
5 CAPRYOL 90
LABRAFIL M
Objectives: Development of nano-emulsion based drug delivery as combined drug loaded form for the possible management of meningitis. Intranasal delivery of nano-emulsion. Study the efficacy and effect of the same in in vitro and in vivo models. To assess the effect of polyphenols on the recommended dosage.
4
5
2.5
CAPRYOL 90 OLEIC ACID LABRAFIL M LAUROGLYCOL ISO PROPYL MYRISTATE TRANSCUTOL LABRASOL
SOLUILITY OF RESVERATROL
14
12
5
CASTOR OIL CAPMUL MCM CORN OIL
SOLUBILITY OF RIFAMPICIN Formulati J8
G9
A9
A7
131.1
96.50
103.9
92.48
PDI
0,357
0.163
0.187
0.224
Zeta
-2.90
-2.86
-2.60
-2.86
on Particle size
MATERIALS AND METHODS
potential
PTPD of selected formulation /1.084
/0.960
1:Isonicotinic acid 137.90>121.10(+) CE: -21.0
2:Resveratrol 228.90>107.00(+) CE: -26.0
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/0.960
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Nasal Toxicity Studies: The nasal toxicity studies was carried out using goat mucus membrane.
Invivo studies: Animal studies were carried out using female wistar rats. The animals were administered with the formulation intranasally and the brains were minced and were subjected to LCMS
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LCMS OF RIFAMPICIN +RESVERATROL
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2500
Cell-line studies: C-6 cell-line of rat glioma was used for finding the toxicity of the drug when targeted to the brain.
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5000
Thermodynamic Stability Studies: Heating Cooling Cycle, Centrifugation and Freeze Thaw Cycle.
1.0
12500
12500
Preparation of drug loaded nanoemulsion: the nanoemulsion was prepared using spontaneous emulsification method. The formulations which had less particle size and zeta potential were selected from the results obtained from pseudoternary phase diagrams.
0.5
2:Resveratrol 228.90>107.00(+) CE: -26.0
2:Resveratrol 228.90>107.00(+) CE: -26.0
CONSTRUCTION OF PSEUDOTERNARY PHASE DIAGRAM: Pseudoternary phase diagrams of various ratios of oil: Smix were constructed. It was done by using CHEMIX software.
LCMS GRAPH OF ISONIAZID + RESVERATROL
700000
/0.960
PREFORMULATION STUDIES: • Construction of Calibration curve: Calibration Curve of isoniazid, Rifampicin and resveratrol were constructed and the linearity was observed. • Compatibility Studies: FTIR and DSC of the compounds was carried out to find out the purity and presence of the functional groups. • Screening of oils: Rifampicin and resveratrol were dissolved in small quantities part by part in various oils. • Screening of surfactants and co-surfactants: The surfactant and co-surfactant which were compatible with the oil were selected.
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Conclusion The lipid based nanoemulsions were prepared by spontaneous emulsification technique. The stability of the formulations was found out by conducting thermodynamic stability studies as well as characterization of the nanoemulsion. From the results obtained that surfactants and co-surfactant played an important role on the physical property of the nanoemulsions. The formulations J8, G9 , A9 and A7 which had a fixed quantity of oil mixed had produced thermodynamically stable product. From these formulations A7 was selected as the optimised formulation as it has the least particle size. And the PDI produced has a narrow particle size distribution and the zeta potential was also within the range. Then these formulations were subjected for cell=line studies and the IC-50 values of the drugs and the formulations were found out. Then, the nasal irritancy test was carried out for both the formulations using the goat mucus membrane. The report states that there was an occurrence of slight inflammation. Then these formulations were administered to animals intranasally. Then, the animals were euthanised and the amount of drug entering the brain was found by using LC-MS/MS. Hence, it was found that 82.69% of Isoniazid+ Resveratrol nanoemulsion and 78.48% of rifampicin + Resveratrol nanoemulsion had crossed the BBB through the olfactory region to the brain. Hence, it can be concluded that maximum amount of drug had entered the brain when administered intranasally.
References Ravindra K Garg, Amita Jain, Hardeep S Malhotra, Avinash Agrawal and Rajiv Garg: 'Drug-resistant tuberculous meningitis', Expert Review Anti Infective Therapy 2013; 11(6), pp. 605-621. Mohammad Nasiruddin, Md. Kausar Neyaz, and Shilpi Das: 'Nanotechnology-Based Approach in Tuberculosis Treatment', Tuberculosis research and treatment 2017; 5(12),