Formulation and Evaluation of Pulsatile s1

FORMULATION AND EVALUATON OF VITAMIN B12 BUCCAL PATCHES

Synopsis for M.Pharm Dissertation submitted to the Rajiv Gandhi University of Health Sciences, Bengaluru, Karnataka.

Ms. KUSHALI JAIN D. M.Pharm. Part - I

Under the guidance of

Dr. GANESH N.S., M.Pharm., Ph.D., ASSOCIATE PROFESSOR DEPARTMENT OF PHARMACEUTICS.

SARADA VILAS COLLEGE OF PHARMACY KRISHNAMURTHYPURAM MYSORE- 570004 2013 - 2014 RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BENGALURU, KARNATAKA. ANNEXURE II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. NAME OF THE CANDIDATE KUSHALI JAIN D. AND ADDRESS (IN BLOCK M.PHARM., PART–I, LETTERS) DEPARTMENT OF PHARMACEUTICS, SARADA VILAS COLLEGE OF PHARMACY, K.M.PURAM, MYSORE - 04 2. NAME OF THE INSTITUTION SARADA VILAS COLLEGE OF PHARMACY, MYSORE - 04 3. COURSE OF STUDY AND SUBJECT MASTER OF PHARMACY IN PHARMACEUTICS 4. DATE OF ADMISSION OF COURSE 14-10-2013 5. TITLE OF TOPIC FORMULATION AND EVALUATION OF VITAMIN B12 BUCCAL PATCHES

6. BRIEF RESUME OF THE INTENDED WORK 6.1 Need for the study ENCLOSURE - I 6.2 Review of the literature ENCLOSURE - II 6.3 Objectives of the study ENCLOSURE – III

7. MATERIALS AND METHODS

7.1 Source of data ENCLOSURE – IV

7.2 Method of collection of data ENCLOSURE - V

7.3Does studies require any ENCLOSURE – VI investigations or interventions to be conducted on patients or other human or animal? If so, please describe briefly.

7.4 Has ethical clearance been obtained ENCLOSURE – VI from your institution in case of 7.3

8. LIST OF REFERENCES ENCLOSURE – VII 9. SIGNATURE OF CANDIDATE

10. REMARKS OF GUIDE RECOMMENDED

11. NAME AND DESIGNATION OF

11.1 Guide Dr.GANESH N.S., M. Pharm., Ph.D., ASSOCIATE PROFESSOR , DEPARTMENT OF PHARMACEUTICS SARADA VILAS COLLEGE OF PHARMACY K.M.PURAM, MYSORE – 04, KARNATAKA.

11.2 Signature

11.3 Co guide (if any) Not applicable

11.4 Signature Not applicable

11.5 Head of department Dr. C. JAYANTHI, M.Pharm., Ph.D., PROFESSOR AND HEAD, DEPARTMENT OF PHARMACEUTICS, SARADA VILAS COLLEGE OF PHARMACY, K.M.PURAM, MYSORE - 04 KARNATAKA.

11.6 Signature

12.1 Remarks of the Principal RECOMMENDED & FORWARDED

12.2 Name and designation of principal Dr. K. HANUMANTHACHAR JOSHI M.Pharm., Ph.D., PRINCIPAL, SARADA VILAS COLLEGE OF PHARMACY, K.M.PURAM, MYSORE - 04.

12.3 Signature 6.0 BRIEF RESUME OF THE INTENDED WORK

ENCLOSURE – I

6.1 Need for the study

Vitamin B-12 deficiency and depletion are common in wealthier countries, particularly among the elderly, and are most prevalent in poorer populations around the world1. Deficiencies of folate or of vitamin B-12 are widespread and constitute major global burden of morbidity that affect all age groups. Vitamin B12 is used for treating and preventing vitamin B12 deficiency, a condition in which vitamin B12 levels in the blood are too low. It is also used to treat pernicious anemia, a serious type of anemia that is due to vitamin B12 deficiency and is found mostly in older people. For this purpose, people use either a supplement that is taken by mouth or a gel that is applied inside the nose. Vitamin B- 12 deficiency alone causes elevation of plasma methylmalonic acid .Other conditions can cause elevations of either homocysteine alone (hypothyroidism, vitamin B-6 deficiency), methylmalonic acid alone (intestinal microbial overgrowth), or both (renal failure). Absent other confounding causes, elevation of plasma homocysteine or methylmalonic acid provides a sensitive indication of folate or vitamin B-12 deficiencies2.

Vitamin B12 is also used for memory loss; Alzheimer’s disease; boosting mood, energy, concentration and the immune system; and slowing aging. It is also used for heart disease, lowering high homocysteine levels (which may contribute to heart disease), male infertility, diabetes, sleep disorders, depression, mental disorders, weak bones (osteoporosis), swollen tendons, AIDS, inflammatory bowel disease, asthma, allergies, a skin disease called vitiligo etc. people use vitamin B12 for amyotrophic lateral sclerosis (Lou Gehrig’s disease), multiple sclerosis, preventing the eye disease, age-related macular degeneration (AMD), Lyme disease and gum disease. It is also used for ringing in the ears, bleeding, liver and kidney disease, and for protection against the poisons and allergens in tobacco smoke3.

B12 is also unusual in that it is dependent upon a second substance, called intrinsic factor, to make its way from the “GI” tract (gastrointestinal tract—the stomach and intestines) into the rest of the body. Without intrinsic factor, which is a unique protein made in the stomach; vitamin B12 cannot gain access to the rest of the body where it is needed. Because of the poor oral bioavailability of the vitamin, its intramuscular (i.m.) injection form is used frequently in the therapeutic practice. The daily dose of the vitamin B12 is only 0.4-2.8 μg and a healthy human liver can accumulate sufficient quantity for two entire years4.

The cyanocobalamin has a large molecular weight (M=1355.7 g/mol), but based on clinical studies it can get through the mucous membranes by passive diffusion, which could enable the possible noninvasive applications of the Vitamin B12. The application of buccal dosage forms has many advantages, as buccal route can be used for systemic delivery because the mucosa has a rich blood Supply and it is relatively permeable. The main objective of this study is to enhance the bio availability of vitamin B12, and this route of drug delivery is of special advantages, including the bypass of first pass effect and the avoidance of presystemic elimination within the GIT. By the use bioadhesive polymeric systems the proper adhesivity to the mucosa can be assured, thus the absorption of the drug through the membrane will be supported5.

ENCLOSURE – II

6.2 Review of the literature

1. David Nowotnik et al., formulated cobalamine nano particles for targeted delivery of nano particles and concluded that vitamin B12 attached to polymer and nano particles provides structures for intracellular drug delivery, using the natural receptor mediated uptake mechanism and Trojan horse strategy. This approach has been successful in the oral drug delivery of peptides which otherwise have little or no oral bioavailability. Vitamin B12 constructs with cytotoxic drugs improve tumor growth inhibition compared with free drug and constructs without vitamin B126.

2. Dorte L. Lildballe et al., studied on maximum load of the vitamin B12 transport system; study on mice treated for four weeks with high dose 0f vitamin B 12 or cobinamide, and study shows that mice are a suitable model for studies of the transport and effect of B12 conjugates. Furthermore, our study emphasizes the importance of monitoring B12 metabolism during treatment with B12 conjugates or analogues7.

3. Carol L Zeuschner et al., studied on vitamin B12 in vegetarian diets and concluded that Vitamin B12 deficiency is a potential concern for anyone with insufficient dietary intake of vitamin B12, including those adhering to a vegan or vegetarian diet or significantly restricting animal-based foods. Studies have found that vegetarians, particularly vegans, have lower serum vitamin B12 levels, and it is likely that anyone avoiding animal-base foods will eventually become deficient if their diet is not supplemented4.

4. Sarath chandran C et al., formulated Buccal patches of Bisoprolol Fumarate using various polymer like 2% chitosan, HPMC, and sodium CMC ,as a polymer and concluded that 2% chitosan may be the best polymer to develop a stable mucoadhesive patches to deliver bisoprolol fumarate. Design and development of such buccal patches may be highly beneficial which can deliver a drug up to 12hrs8.

5. Surya N et al., Prepared and evaluated Buccal Patches for Delivery of Atenolol using sodium alginate with various polymer like carbapol 934 p, sodium carboxy methyl cellulose and hydroxyl propyl methyl cellulose in various proportion and combination were fabricated using solvent casting technique and showed satisfactory physicochemical and mucoadhesive characteristics9.

6. Chinna Reddy P et al., studied on bioadhesive drug delivery system current status of formulation and evaluation methods and concluded that buccal adhesive system offer innumerable advantages in terms of accessibility administration and withdrawal, retentivity, low enzymatic activity, economy and high patient compliance. Adhesions of these drug delivery devices to mucosal membranes lead to an increased drug concentration gradient at the absorption site and therefore improve bioavailability of systemically delivered drugs10.

7. Mona Semalty et al., had formulated and evaluated mucoadhesive buccal films of Glipizide using mucoadhesive polymers HPMC-E15, CP-934P, Eudragit RL-100 and sodium CMC. Propylene glycol was used as the plasticizer as well as penetration enhancer. And study indicated a good potential of erodible mucoadhesive buccal films containing glipizide for systemic delivery with an added advantage of circumventing the hepatic first pass metabolism. The results of the study show that therapeutic levels of glipizide can be delivered buccally11. 8. Y Indira Muzib et al., formulated mucoadhesive buccal films of Glibenclamide usind HPMC3000 From the results it can be concluded that HPMC3000 at low concentrations can be useful for buccal delivery of glibenclamide in a controlled manner. Buccal drug delivery is a promising area for continued research with the aim of systemic delivery of orally inefficient drugs as well as a feasible attractive alternative for noninvasive delivery of potent peptide and protein drug molecules. However, the need for safe and effective buccal permeation/absorption enhancers is a crucial component for a prospective future in the area of buccal drug delivery12.

9. Santhosh Kumar Mishra et al., studied on development and evaluation Mucoadhesive buccal patches of Flurbiprofen and study suggested that It was found that formulation containing HPMC with β-CD had better release profile than that of other formulations. Higher value of R and lowest value of RSS was seen in the Peppas model. Hence the best fit model was that of Peppas. HPMC, PEG-600 and the drug-β-CD binary system containing patches have good flexibility, convenient thickness, prolonged in vitro residence time, good swelling properties and higher value of percentage drug release for a prolonged period of time. Hence, it was found to be an effective system for enhancing bioavailability and avoid GIT side effects and bypass liver enzymes13.

ENCLOSURE – III

6.3 Objectives of the study The objectives of present study are as follows: 1. To carry out pre-formulation studies for possible drug /polymer/excipients interactions using FTIR. 2. To design and develop mucoadhesive buccal patches of vitamin B12 using various polymer. 3. To characterize the patches. 4. To carry out various evaluaion studies as mentioned below. 5. In vitro diffusion / release studies. 6. In vitro release kinetics. MATERIALS AND METHODS:

DRUG : Vitamin B12 POLYMERS: Using various suitable polymers like carbapol 934 p, sodium carboxy methyl cellulose and hydroxyl propyl methyl cellulose, chitosan, sodium alginate, guar gum, casein etc. SOLVENTS : Ethanol, acetic acid, phosphate buffer (saline-pH 6.8) etc.

METHODS

1. Preparation  Preformulation Studies  Formulation of buccal patches of vitamin B12  Solvent casting technique (method)  FTIR and DSC studies

2. Evaluation  Film thickness  Drug content  Folding endurance  Measurement of surface pH  Swelling behavior  Residence time (Ex vivo mucoadhesion time)  Drug release studies  Release kinetics ENCLOSURE – IV 7.0

7.1. Source of data

a. Library: Sarada Vilas College of Pharmacy.

b. E-library: Sarada Vilas College of Pharmacy.

ENCLOSURE – V

7.2. Method of collection of data

1. Compatibility studies between the drug, polymers and penetration enhancers will be carried out using IR and DSC instruments. 2. Patches will be designed using different polymers like carbapol 934 p, sodium carboxy methyl cellulose and hydroxyl propyl methyl cellulose, 2% chitosan etc. and plasticizers such as propylene glycol, glycerin etc adopting suitable technique. 3. Physical characterization like appearance, thickness, tensile strength, absorption etc. will be carried out adopting suitable method. 4. Drug release studies will be carried out in simulated physiological fluids using Keshary- chein or franz diffusion cell.

ENCLOSURE – VI

7.3. Does the study require any investigation or intervention to be conducted on patients or other humans or animals? If so, please mention briefly. - No - 7.4. Has ethical clearance been obtained from your institution in case of 7.3? - NA- 8.0 ENCLOSURE – VII

LIST OF REFERENCES:

1. Lindsay H Allen. How common is vitamin B-12 deficiency. Am J Clin Nutr. 2009;89:693–96. 2. Ralph green.Indicators of assessing Folate and VitaminB12 status and monitoring the efficacy of intervention strategies. Am J Clin Nutr. 2011;94:666-72. 3. Mc Elnay JC, Swarbick J, Boylan JC. Encyclopedia of pharmaceutical technology. 2nd ed. New York: Marcel Dekkar; 1990;189. 4. Carol L Zeuschner, Bevan D Hokin, Kate A Marsh, Angela V Saunders, Michelle A Reid Melinda R Ramsay, Vitamin B12 and vegetarian diets.MJA. 2012;27-32. 5. Barnabas Szaboa.b, Nikolett Kallaic, Gergo Tothd, Gergely Hetenyia, Romana Zelkoa. Drug release profiles and microstructural characterization of cast and freeze dried vitamin B12 buccal films. Journal of Pharmaceutical and Biomedical Analysis 2013;89:83-87. 6. David Nowotnik. Cobalamine nanoparticles for targeted delivery of RNAi therapeutics. Access pharm. 2003;5:1-18. 7. Dorte L. Lildballe, Elena Mutti, Henrik Birn, Ebba Nexo. Maximal Load of the Vitamin B12 Transport System: A Study on Mice Treated for Four Weeks with High-Dose Vitamin B12 or Cobinamide, PLOS ONE. 2012;7:10. 8. Sarath chandran C, KV. Shijith, KV . Vipin and Ann Rose Augusthy. Formulation Buccal patches of Bisoprolol Fumarate using various polymers by solvent casting technique. IJPCBS 2013;3(3):854-60. 9. Surya N. Ratha Adhikari, Bhabani S. Nayak, Amit K. Nayak, and Biswaranjan Mohanty. Formulation and evaluation of buccal patches for delivery of atenolol. AAPS PharmSciTech.2010;11(3). 10. Chinna Reddy P, Chaitanya K.S.C., Madhusudan Rao Y. By "Tehran University of Medical Sciences , studied on bioadhesive drug delivery system current status of formulation and evaluation methods. DARU Journal of Pharmaceutical Sciences 2011;19(6):385-403. 11. Mona semalty, A.semalty and G.kumar. Mucoadhesive buccal films of glipizide. Indian J Pharm Sci.2008;70(1):43–48. 12. Muzib Y I, Kumari K S. Mucoadhesive buccal films of glibenclamide: Development and evaluation. Int J Pharma Investig 2011;1:42-7. 13. Santhosh kumar mishra, Navneet garud and Ranjit singh. Development and Evaluation of mucoadhesive buccal patches of Flurbiprofen. Acta Poloniae Pharmaceutica n Drug Research. 2011; 68(6):955-64.