Rajiv Gandhi University of Health Sciences s174

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

KARNATAKA, BANGALORE

M. PHARM SYNOPSIS

YEAR 15/05/2010

TITLE OF THE SYNOPSIS

FORMULATION AND EVALUATION OF METHOTREXATE DOSAGE FORM FOR COLON SPECIFIC DRUG DELIVERY

ASHIF ANJUKANDAN

M.PHARM, PART- I

DEPARTMENT OF PHARMACEUTICS,

UNDER THE GUIDANCE OF

Mrs. SELVI ARUN KUMAR M. Pharm

Asst. Professor

DEPARTMENT OF PHARMACEUTICS

INSTITUTION

GAUTHAM COLLEGE OF PHARMACY

R. T. NAGAR, BANGALORE-32, KARNATAKA RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BANGALORE, KARNATAKA ANNEXURE – II PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION

1. NAME OF THE CANDIDATE Mr. ASHIF ANJUKANDAN AND ADDRESS DEPARTMENT OF PHARMACEUTICS, GAUTHAM COLLEGE OF PHARMACY, SULTAN PALAYA, R.T NAGAR (PO) BENGALURU – 32, KARNATAKA.

PERMANENT ADRESS S/o ABDUL AZEEZ A.K. ANJUKANDAN HOUSE VALIYORA (P.O), 676304 (Pin) VENGARA, MALAPPURAM (Dist). KERALA. 2. NAME OF THE INSTITUTION DEPARTMENT OF PHARMACEUTICS, GAUTHAM COLLEGE OF PHARMACY, SULTAN PALAYA, R.T NAGAR (PO) BENGALURU – 32, KARNATAKA.

3. COURSE OF STUDY AND MASTER OF PHARMACY IN SUBJECT PHARMACEUTICS

DATE OF ADMISSION TO 4. COURSE 15/05/2010

5. TITLE OF THE TOPIC “FORMULATION AND EVALUATION OF METHOTREXATE DOSAGE FORM FOR COLON SPECIFIC DRUG DELIVERY”

2 6. BRIEF RESUME OF THE INTENDED WORK:

6.1 NEED FOR THE STUDY:

The word new or novel in the relation to drug delivery system is a search for something out of necessity. An appropriately designed sustained or controlled release drug delivery system can be major – advance towards solving the problem associated with the existing drug delivery systems.1

Colon-specific drug delivery system has gained increased importance not just for the delivery of the drugs for treatment of local diseases associated with the colon but also for its potential for the delivery of proteins and therapeutic peptides. The following different categories of drugs which require local delivery are suitable for colon drug delivery:      Drugs to treat irritable bowel syndrome. e.g. Sulfasalazine, steroids like Fludrocortisones. Drugs to treat colonic cancer require local delivery e.g. 5-Fluorouracil, Methotrexate.  Protein and peptide drugs - eliminating drug degradation e.g. Growth hormones, Calcitonin,  To treat infectious diseases (amoebiasis & helminthiasis) - e.g. Metronidazole,     To treat Rheumatoid arthritis (NSAIDS), Nocturnal asthma, Angina require delay in absorption due to Circadian rhythms.  Drugs showing more selective absorption in colon than small intestine due to small extent of paracellular transport e.g. Glibenclamide, Diclofencac, Theophylline, Metoprolol,.

The various strategies for targeting orally administered drugs to the colon include covalent linkage of a drug with a carrier, coating with pH-sensitive polymers, formulation of timed released systems, exploitation of carriers that are degraded specifically by colonic bacteria, bioadhesive systems and osmotic controlled drug delivery systems. 3 Cancer of the colon and rectum is one of the most common internal malignancies. Colorectal cancer is the second-leading cause of cancer deaths in the United States. In 2005, estimated 1, 45290 new cases of colon cancer were diagnosed in the United States. Almost all cases of colorectal cancer begin with the development of benign or noncancerous polyps. When colon cancer cells spread outside the colon or rectum to lymph nodes, they may also spread to other lymph nodes, the liver, or other organs. Surgery is still a mainstay of the treatment of colorectal cancer. 4 Chemotherapy is also used to treat advanced colorectal cancer. But conventional chemotherapy is not as effective in colorectal cancer as it is in other cancers, as the drug does not reach the target site in effective concentrations. Thus, effective treatment demands increased dose size, which may lead to undue consequences. To improve this situation, pharmaceutical technologists have been working on ways to deliver the drug more efficiently to the colon, where it can target the tumor tissues. It is possible that delivery of small quantities of antineoplastic agent to the inner surface of the colon could destroy small tumors that arise spontaneously in this region, reducing the need for surgery.4 Various approaches, namely microcapsules, compressed microcapsules, and modified tablets can be used for the colon specific drug delivery.8 Methotrexate ((2S)-2-[(4-{[(2,4-Diaminopteridin-6-yl)methyl](methyl)amino} phenyl) formamido] pentanedioic acid), is an antimetabolite and antifolate drug used in treatment of cancer, autoimmune diseases and as an abortifacient in the induction of medical abortions. It acts by inhibiting the metabolism of folic acid. It’s used in treatment of Chron’s disease and colorectal cancer. It is also used as Antirheumatic Agents, Immunosuppressive Agents, Dermatologic Agents, Antimetabolite. Methotrexate competitively inhibits dihydrofolate reductase (DHFR), an enzyme that participates in the tetrahydrofolate synthesis.5 Dihydrofolate reductase catalyses the conversion of Dihydrofolate to the active Tetrahydrofolate. Folic acid is needed for the de novo synthesis of the nucleoside thymidine so inhibits the synthesis of DNA, RNA, thymidylates, and proteins. Methotrexate is weak dicarboxylic acid with pKa of 4.8 to 5.5, thus it is mostly ionized at physiological pH and oral absorption is saturated & dose dependent. A mean oral bioavailability of about 33% and mean intra muscular bioavailability is 76%. Methotrexate is metabolized by intestinal bacteria to the inactive metabolite 4-amino-4 deoxy-N- methylpteroic acid (DAMPA) and account for less than 5% loss of oral dose. 6 The aim of this work is to design a Colon Specific Dosage form of Methotrexate. Physicochemical properties and half life of Methotrexate makes its suitable candidate for colonic drug delivery system .The site specific drug delivery of Methotrexate in this formulation reduces the adverse effects and fluctuation in plasma concentration.

6.2 REVIEW OF LITERATURE

Literature review for undertaking the study was done by referring to various national and international journals, published articles in various official standard books and referring to various websites on the internet. M.NAJMUDDIN et al.,1 Prepared Flurbiprofen microcapsules by solvent evaporation method using Eudragit L‐100 and Eudragit S‐100 mixture [1:2, 1:3, 1:4, 1:5] were used as 4 pH‐sensitive polymers and evaluated for various physicochemical parameters such as particle size, percentage yield, incorporation efficiency, drug polymer compatibility (IR study), scanning electron microscopy (SCM) and drug release of microcapsule (pH 6.8) for 12 hrs. Result shows that as the concentration of polymer increases it affects the particle size, percentage yield and drug release of micro capsules. Results of this study suggest that Flurbiprofen microcapsules can be successfully designed to develop sustained drug delivery, which can improve compliance by reducing dosing frequency. R. Saraswathi et al.,7 Developed and evaluated Albendazole microcapsules for colon- specific delivery for better treatment of Helminthiasis, Filariasis, and colorectal cancer by avoiding the side effects. Microcapsules prepared by the use of different concentrations of sodium alginate and guar gum and studied for entrapment efficiency, surface morphology and particle size analysis. In vitro drug release study in presence and absence of cecal content were also studied. The particle size range is maximum for microcapsules with high concentration of sodium alginate and guar gum. The results of in vitro release study indicated that the amount of drug release decreased significantly with an increase in guar gum concentration but it was reverse with respect to sodium alginate concentration. D. Nagasamy Venkatesh et al.,8 Developed Tegaserod maleate colon specific drug delivery systems via the oral route for irritable bowel syndrome, using Eudragit L 100 and S100 mixture (1:1, 1:2, and 1:3) as pH-sensitive polymers. Various approaches, namely microcapsules, compressed microcapsules, and modified tablets were made for this study. The microcapsules were prepared by the emulsion-solvent evaporation method using drug and mixture of polymers and evaluated for various physicochemical parameters such as particle size, surface morphology, drug loading capacity, and in vitro dissolution studies by half-dilution method employing various pH environments (pH 1.2-6.8) for 24 hours. The batch prepared using the 1:2 drug polymer ratio was selected as an ideal batch for compression to get the compressed tablet of the microcapsules. The modified tablets were also prepared using the drug with hydroxylpropylmethylcellulose as the inner material and ethyl cellulose as the outer material employing the double compression technique. Comparisons of microcapsules, compressed microcapsules, and modified tablets containing tegaserod maleate indicated that the drug release profiles from the microcapsules were found to be better than the compressed microcapsules and the modified tablets in the colonic environment. Mohini Chaurasia et al.,9 Prepared and characterized guar gum microspheres containing methotrexate (MTX) for local release of drug in the colon, which is a prerequisite for the effective treatment of colorectal cancer. Guar gum microspheres were prepared by the emulsification method using glutaraldehyde as a cross-linking agent. Microsphere were evaluated particle size, shape, and surface morphology. MTX-loaded microspheres demonstrated high entrapment efficiency. The in vitro drug release was investigated using a US Pharmacopeia paddle type (type II) dissolution rate test apparatus in different media (phosphate-buffered saline [PBS], gastrointestinal fluid of different pH, and rat cecal content release medium), which was found to be affected by a change to the guar gum concentration and glutaraldehyde concentration. Lanjhiyana Sanjay Kumar and Dangi Jawahar Singh.,10 Developed pulsatile release based enteric coated capsules for site specific delivery of Methotrexate to colon. The modified formulation was capable of delaying onset of drug release for a programmed lag time period of 3-5 h in the simulated physiological environment of upper gastrointestinal tracts depends on coating ratios of HPMC (inner coating) and Eudragit-S100 (outer coating). Dissolution studies demonstrates that, polymeric coated formulations of HPMC and pH sensitive Eudragit-S 100 were gastro-resistance for 2 h at pH 1.2 and further for 3 h at pH 6.8, since they released only 3-7 % of drug in physiological environment of stomach and small intestine. The complete disintegration occurred only after a certain lag time of 4 to 5 h. DSC thermogram analysis shows no possibility of interaction between drug and polymers used in the study. The study result demonstrated that the developed system can be a promising device for targeting of methotrexate to colonic region for treatment of colorectal cancer Parul Trivedi et al.,11 Developed formulation where in Aceclofenac is microencapsulated with Eudragit (S 100, RL 100, and RS 100), using an O/W emulsion-solvent evaporation technique and evaluated micromeritic properties including angle of repose, bulk density, tapped density, Carr's Index, Hausner's ratio, and particle size determination and also drug loading, in vitro drug release as well as for scanning electron microscopy. In vitro drug release studies were carried out up to 24 h in three different pH media, i.e., 0.1 N HCl (pH 1.2), phosphate buffer (pH 6.8), and phosphate buffer (pH 7.4). Wu Ping et al.,12 Developed a chitosan-methotrexate covalently conjugated nanoparticles (CS-MTX-TPP NPs) as a potential delivery system for Methotrexate (MTX). MTX was first conjugated to CS by using Glutaraldehyde as cross-linked agent, and followed by the process of ionic gelation between MTX-conjugated CS and Sodium Tripolyphosphate (TPP) to form CS-MTX-TPP NPs at mild reaction conditions. Nanoparticles evaluated for the encapsulation efficiency of MTX. The characterizations by Atomic Force Microscopy and Photon Correlation Spectroscopy showed the CS-MTX-TPP NPs had a spherical shape and good dispersion with diameter of sub-200-nm and zeta potential of 30 mV. Additionally, in vitro release test revealed that the stable covalent bonding of CS and MTX was beneficial for providing slow release for MTX. 6 Mukesh Ukawala et al.,13 Formulated and characterized calcium phosphate nanoparticle containing anticancer drug, Methotrexate (MTX). Calcium Phosphate nanoparticles containing MTX (CaPi-MTX) were prepared by reverse micelles technique and evaluated average size and entrapment efficiency. The IR spectrum of CaPi-MTX showed characteristics of composite formation of hydroxyapatite with MTX. X-RD analysis revealed that, CaPi-MTX nanoparticles were crystalline and in hydroxyapatite form. TEM studies showed that CaPi-MTX nanoparticles were spherical in shape. In vitro release study of CaPi-MTX nanoparticles showed slow release of MTX at physiological pH (pH 7.4) while >90% release was observed within 3-4 hours at endosomal pH (pH 5.5 and pH 6.0). Confocal microscopy was performed using CHO cell lines, showed intracellular localization of FITC-Dextran loaded calcium phosphate nanoparticles. Results indicate that prepared CaPi-MTX nanoparticles could serve the purpose for intracellular drug delivery. M. Chaurasia et al.,14 Prepared calcium pectinate (Ca-pectinate) microspheres to deliver methotrexate in the environment of colon by modified emulsification method using calcium chloride as cross linker. All the formulations were evaluated for various physicochemical parameters like Particle size of the microspheres, Encapsulation efficiency and in vitro drug release studies were performed in simulated gastric fluid for 2 hours and simulated intestinal fluid for 3 hours. In vitro release rate studies were also carried out in simulated colonic fluid in presence of rat caecal contents. Moreover, release rate studies were also carried out after enzyme induction by treating the rats with 1 ml of 1% w/v aqueous dispersion of pectin for 7 days. This study concluded that calcium pectinate microspheres can be used to effectively localize the release of drug in the physiological environment of colon. Ziyaur Rahman et al.,15 Prepared and evaluated the colon-specific microspheres of 5- Fluorouracil for the treatment of colon cancer. Core microspheres of alginate were prepared by the modified emulsification method in liquid paraffin and by cross-linking with calcium chloride and were coated with Eudragit S-100 by the solvent evaporation technique to prevent drug release in the stomach and small intestine. The microspheres were characterized by shape, size, surface morphology, size distribution, incorporation efficiency and In vitro drug release studies. Release was sustained for up to 20 hours in formulations with core microspheres to a Eudragit S-100 coat ratio of 1:7, and there were no changes in the size, shape, drug content, differential scanning calorimetry thermogram, and In vitro drug release after storage at 40°C/75% relative humidity for 6 months. V.S. Mastiholimath et al.,16 Investigated an oral colon specific, pulsatile device to achieve time and/or site specific release of Theophylline, based on chronopharmaceutical consideration. They filled Eudragit microcapsules of Theophylline insoluble hard gelatin capsule body and sealed with a hydrogel plug. The entire device was enteric coated to overcome the variability in gastric emptying time and to achieve a colon-specific release . Microcapsules were prepared in four batches, with Eudragit L-100 and S-100 (1:2) by varying drug to polymer ratio and evaluated for the particle size, drug content and In vitro release profile and from the obtained results; one better formulation was selected for further fabrication of pulsatile capsule. Different hydrogel polymers were used as plugs, to maintain a suitable lag period and it was found that the drug release was controlled by the proportion of polymers used. In vitro release studies of pulsatile device revealed that, increasing the hydrophilic polymer content resulted in delayed release of Theophylline from microcapsules. The gamma scintigraphic study pointed out the capability of the system to release drug in lower parts of GIT after a programmed lag time for nocturnal asthma. Programmable pulsatile, colon-specific release has been achieved from a capsule device over a 2–24 h period, consistent with the demands of chronotherapeutic drug delivery. S. K. Uma Devi et al.,17 Developed novel colon specific drug delivery systems for Aceclofenac using pectin as a microbially degradable polymeric carrier and to coat the optimized batches with a pH dependent polymeric coating solution containing Eudragit L 100 and S 100 (1:4). Pellets containing four proportions of pectin were prepared. The pellets were evaluated for physicochemical properties, drug content, dissolution, water uptake & erosion characteristics, In vitro drug release studies. The dissolution profile and in vitro release kinetics showed that pectin pellets were promising for controlled delivery of the drug. The findings of this study concluded that pectin pellets are promising for colon targeting of Aceclofenac to synchronize the chronobiological symptoms for effective treatment of Rheumatoid arthritis. R.S.S. Shendge et al.,18 Prepared Colon targeted drug delivery system by using Dextrin, polysaccharide, as a carrier for Aceclofenac matrix tablet by wet granulation technique . Different binder like Ethyl cellulose, Sodium CMC and Sucrose were used during preparation of matrix tablets containing dextrin and various excipients. Evaluation was done by different IPQC tests, content uniformity and in vitro drug release study. Drug release profile was evaluated in simulated gastric, intestinal fluid and simulated colonic fluid. The matrix tablet containing binder system of ethyl cellulose and dextrin as a carrier was found to be suitable for targeting the colon as compare to other matrix tablets containing different binders because of fewer amounts of drug release in the simulated gastric and intestinal fluid. P.B. Aswar et al.,19 Developed colon targeted matrix tablet of Diclofenac Sodium by using various proportions of gaur gum as a carrier and Sodium-CMC, Sucrose 70% and Ethyl cellulose as a binder by wet granulation technique. The prepared matrix tablets were evaluated by different In-Process Quality Control tests like content uniformity and drug 8 release study and Drug release profile in simulated gastric fluid, intestinal fluid and simulated colonic fluid. The matrix tablets containing gaur gum as a carrier and ethyl cellulose as a binder was found to be suitable for targeting Diclofenac Sodium for local action in the colon as compared to other matrix tablets containing different binders because of amount of drug release in simulated gastric fluid and intestinal fluid.

6.3 OBJECTIVES OF THE STUDY 1. Development of suitable dosage form based on the Preformulation studies 2. Evaluation of formulated Dosage form for following parameters. i) Percentage yield ii) Incorporation efficiency iii) Particle Size iv) Angle of Repose v) Morphology and Surface Appearance vi) Infrared Spectroscopy vii) In – Vitro drug release.

7. MATERIALS AND METHODS Drug : Methotrexate Polymer : Eudragit L100 or Eudragit S100, Eudragit RS 100, Eudragit RSPO, Cellulose Acetate Phthalate, Shellac and Guar Gum. Method :Solvent evaporation and other innovative techniques.

7.1 SOURCE OF DATA:  Available published literatures  Lab experiment.

7.2 METHOD OF COLLECTION OF DATA: Data on drugs and polymers will be collected by literature survey. Data on Preformulation and post formulation studies will be collected through experimental and analytical methods.

7.3 Does the study require any investigations or invention to be conducted on patients or other human or animals? If so, please mention briefly. NA

7.4 Has ethical clearance been obtained from your institution in case of 7.3?

8. REFERENCES

1. M. Najmuddin, Vishal Patel, Aejaz Ahmed, S. Shelar, T. Khan. “Preparation and

Evaluation of Flurbiprofen Microcapsules for Colonic Drug Delivery System”. Int.

J. Pharmacy Pharm. Sci. 2010; 2(2): 83-87.

2. Sateesh Kumar Vemula and Prabhakar Reddy Veerareddy. “Different approaches to

design and evaluation of colon specific drug delivery systems”. Int.J.Pharmacy and

Tech. 2009; 1 (1): 1-35.

3. M.K. Chourasia, S.K. Jain. “Pharmaceutical approaches to colon targeted drug

delivery systems”. J. Pharm. Pharm. Sci. 2003; 6(1): 33-66.

4. M. Chaurasia, M.K. Chourasia, N.K. Jain, A. Jain, V. Soni, Y.Gupta, S.K. Jain.

“Cross-Linked Guar Gum Microspheres: A Viable Approach for Improved Delivery

of Anticancer Drugs for the Treatment of Colorectal Cancer”. AAPS PharmSciTech.

2006; 7(3): Article 74.

5. P. T. Ravi Rajagopalan, Zhiquan Zhang, Lynn McCourt, Mary Dwyer, Stephen J.

Benkovic, and Gordon G. Hammes . "Interaction of dihydrofolate reductase with

methotrexate: Ensemble and single-molecule kinetics". PNAS. 99(21): 13481–13486.

6. Meyer. L.M., Miller F.R., Rowen M.J., Bock. G., Rutzky. J. (1950). "Treatment of

acute leukemia with amethopterin (4-amino, 10-methyl pteroyl glutamic acid)".

Acta Haematologica. 4 (3): 157–67.

10 7. R. Saraswathi, S.P.Simi, C.Sankar, P.N. Krishnan, C. Dilip, K. Ameena. “Guar gum

based microcapsules for colonic delivery of albendazole: Development and In-vitro

evaluation”. Res. J. Pharm. Biol. & Chem. Sci. 2010; 1(4): 373.

8. D. Nagasamy Venkatesh, Ajay Kumar Reddy, M.K. Samanta, B. Suresh.

“Development and in vitro evaluation of colonic drug delivery systems for

Tegaserod maleate”. Asian J. Pharmaceutics. 2009; 3 (1): 50-53.

9. M. Chaurasia, M.K. Chourasia, N.K. Jain, A. Jain, V. Soni, Y. Gupta, S.K. Jain.

“Cross-Linked Guar Gum Microspheres: A Viable Approach for Improved Delivery

of Anticancer Drugs for the Treatment of Colorectal Cancer”. AAPS PharmSciTech.

2006; 7(3): Article 74.

10. Lanjhiyana Sanjay Kumar and Dangi Jawahar Singh. “Development and In-Vitro

Drug Release Studies of Methotrexate from Modified Pulsatile Release Guar Gum

based Enteric Coated Capsules for Colon Specific Delivery”. Indian J.Pharm. Educ.

Res. 2008; 42(2): 154-160.

11. P. Trivedi, A Verma, N. Garud. “Preparation and characterization of aceclofenac

microspheres”. Asian J. Pharmaceutics. 2008; 2: 110-115.

12. Wu Ping, He Xiaoxiao, Wang Kemin, Tan Weihong, He Chunmei, Zheng Mingbin.

“A Novel Methotrexate Delivery System Based on Chitosan-Methotrexate

Covalently Conjugated Nanoparticles”. J. Biomed. Nanotech. 2009; 5(5): 557-564.

13. Mukesh Ukawala, Kulkarni Vijay, Tushar Rajyaguru, R.S.R. Murthy. “Methotrexate

Loaded Self Stabilized Calcium Phosphate Nanoparticles: A Novel Inorganic Carrier

for Intracellular Drug Delivery”. J. Biomed. Nanotech. 2009; 5(1): 99-105.

14. M. Chaurasia, M. K. Chourasia, Nitin, K. Jain, A. Jain, V. Soni, Y. Gupta, S. K. Jain.

“Methotrexate bearing calcium pectinate microspheres: a platform to achieve colon-

specific drug release”. Curr. Drug Deliv. 2008; 5(3): 215-219. 15. Z. Rahman, K. Kohli, R.K. Khar, M. Ali, N.A. Charoo, A.A. Shamsher.

“Characterization of 5-Fluorouracil Microspheres for Colonic Delivery”. AAPS

PharmSciTech. 2006; 7(2): Article 47.

16. V. S. Mastiholimath, P. M. Dandagi, S. Samata Jain, A. P. Gadada and A. R.

Kulkarni. “Time and pH dependent colon specific, pulsatile delivery of Theophylline

for nocturnal asthma”. Int. J. Pharmaceutics. 2007; 328(1): 49-56.

17. S. K. Uma Devi, R. Thiruganesh and S. Suresh. “Preparation and characterization of

pectin pellets of Aceclofenac for colon targeted drug delivery”. J. Chem. Pharm.

Res. 2010; 2(1): 361-374.

18. Raosaheb S. Shendge, Fatima J. Sayyad, Kishor S. Salunkhe and Rasika D. Bhalke.

“Development of Colon Specific drug delivery of Aceclofenac by using effective

binder system of ethyl cellulose”. Int. J. Pharma & BioSci. 2010; 1(3).

19. P. B. Aswar, S. S. Khadabadi, B. S. Kuchekar, T. P. Wane, N. Matake.

“Development and in-vitro evaluation of colon specific formulation for orally

administered Diclofenac Sodium”. Arch. Pharm. Sci. & Res. 2009; 1(1): 48-53.

12 9. Signature of Candidate

10. Remarks of the guide The above information is true to the best of my knowledge and the work will be done under my guidance.

11. 11.1 Name and Designation of Guide Mrs. SELVI ARUN KUMAR. M.Pharm Asst. Professor, Department of Pharmaceutics, Gautham College of Pharmacy, Sultanpalaya, R.T Nagar (P.O) Bangalore – 32, Karnataka.

11.2 Signature

11.3 Co-Guide (IF ANY)

11.4 Signature

11.5 Head of the Department Dr. SANJAY PRAHALAD UMACHIGI. M.Pharm, PhD. Prof. & Head Department of Pharmaceutics, Gautham College of Pharmacy, Sultanpalaya, R.T Nagar (P.O) Bangalore – 32, Karnataka. 11.6 Signature

12. 12.1 Remarks of the Principal The above mentioned information is correct and I recommend the same for approval. 12.2 Signature

Mrs. ARCHANA. P. SWAMY. M. Pharm,( PhD). Principal Gautham College of Pharmacy, Sultanpalaya, R.T Nagar (P.O) Bangalore – 32, Karnataka.