a) BRIEF RESUME OF INTENDED WORK:

6.1. Need for the study:

Conventional oral drug dosage forms such as tablets; capsules provide specific drug concentration in systemic circulation without offering any control over drug delivery and also cause great fluctuations in plasma drug levels. Single unit dosage forms have the disadvantage of a release all or nothing emptying process while the multiple unit particulate system pass through the GIT to avoid the vagaries of gastric emptying and thus release the drug more uniformly. The uniform distribution of these multiple unit dosage forms along the GIT could result in more reproducible drug absorption and reduced risk of local irritation; this gave to oral controlled drug delivery1. A well-designed controlled drug delivery system can overcome some of the problems of conventional therapy and enhance the therapeutic efficacy of a given drug. To obtain maximum therapeutic efficacy, it becomes necessary to deliver the agent to the target tissue in the optimal amount in the right period of time there by causing little toxicity and minimal side effects. There are various approaches in delivering a therapeutic substance to the target site in a sustained controlled release fashion. One such approach is using microspheres as carriers for drugs. Microspheres are characteristically free flowing powders consisting of proteins or synthetic polymers which are biodegradable in nature and ideally having a particle size ranging from 1μm -1000 μm in diameter2. Microspheres can encapsulate many types of drugs including small molecules, proteins, and nucleic acids etc. They are generally biocompatible, can provide high bioavailability, and are capable of sustained release for long periods of time3. However; the success of these microspheres is limited owing to their short residence time at the site of absorption. It could, therefore be advantageous to have means for providing an intimate contact of the drug delivery system with the absorbing membranes. This can be achieved by coupling bioadhesion characters to microspheres and developing mucoahesive microspheres. Mucoadhesion is topic of current interest in the design of drug delivery system. Mucoadhesive drug delivery system is delivery system, which utilizes the property of bioadhesion of certain polymers, which become adhesive on hydration. Mucoadhesive microsphere exhibits a prolonged residence time at the site of application and facilitates an intimate contact with the underlying absorption surface and thus contributes to improved or better therapeutic performance of drug. Mucoadhesive drug delivery systems promises several advantages that arise from localization at a given target site, prolonged residence time at the site of drug absorption and an intensified contact with the mucosa increasing the drug concentration gradient. Hence, uptake and consequently bioavailability of the drug is increased and frequency of dosing reduced with the result that patient compliance is improved4. The objective of drug targeting is to achieve desired pharmacological response at a selected site without undesirable interactions. It is well known that most of anti-inflammatory drugs administered in therapeutic doses, increase the sensitivity of the gastric mucosa against the aggressive factor, and the acid can produce gastro- intestinal inflammations or ulcers5. Drug delivery systems that can encapulate such irritating drugs would be advantageous. Nonsteriodal anti-inflammatory drugs (NSAIDs) are amongst the most commonly prescribed medications in the world. As awareness of the GI side effects associated with NSAIDs increases, safety becomes a primary requisite in treatment. A trend in NSAID development has been to improve therapeutic efficacy and reduce the severity of GI side effects through altering dosage forms by modifying release of the formulations to optimize drug delivery6. Aceclofenac is one of the emerging NSAID anti-inflammatory molecules. It is a phenyl acetic acid derivative [2- (2',6’-dichlorophenyl) amino] phenylacetoxyacetic acid], a novel NSAID indicated for the symptomatic treatment of pain and inflammation. Aceclofenac has a very short biological half-life (about 4 hours) which makes aceclofenac an ideal candidate for prolong release. The high concentration with rapid absorption of aceclofenac causes adverse effect to GIT. The improved therapeutic efficacy of aceclofenac and reduce the severity of upper GI tract side effect, can be achieved by modifying release of the drug by formulating to an controlled mucoadhesive microspheres as drug delivery system. So this project is an attempt to formulate and evaluate mucoadhesive microspheres of ant-inflammatory drug, aceclofenac for site-specific oral controlled drug delivery.

6.2. Review of literature:

 Eudragit coated chitosan–Ca–alginate microparticles loaded with budesonide (BDS) was studied as candidate for oral delivery of BDS, which exhibited controlled release properties, could be used as new therapeutic potential carriers for local treatment of inflammatory bowel disease7  The release of acetazolamide after per oral administration from Eudragit RS to Eudragit RL microspheres were studied. Controlled release profiles were obtained without initial peak levels, which can reduce dosing frequency, decrease side effects and improve patient compliance8.  Mucoadhesive microspheres of acyclovir (ACV-ad-ms) were prepared by emulsion solvent evaporation method. Acv-ad-ms could remain in the stomach for an extended time and ACV be better absorbed in the upper part of gastrointestinal tract and also observed increased oral bioavailability due to the prolonged residence time in stomach9.  The loratadine entrapment efficiency and mucoadhesion property was determined from ethyl cellulose in comparison to chitosan in microspheres. Chitosan ensures longer retention of the drug delivery system at the site of deposition, as loratadine was significantly less present at their surface, and consequently had less influence on bioadhesion. Higher chitosan content (EC/CM 1:3) ensured more compact coating of EC cores of microspheres, improving their bioadhesive properties10.  The preparation and characterization of mucoadhesive microspheres with Famotidine as model drug for prolongation of gastric residence time was carried out. The microspheres were prepared by the w/o emulsification solvent evaporation method using mucoadhesive polymers sod. CMC and a release controlling polymer sodium alginate. The prepared microspheres exhibited prolonged drug release11.  Amoxicillin trihydrate mucoadhesive microspheres were prepared with ethyl cellulose as a matrix and carbopol 934P as a mucoadhesive polymer. In conclusion, the prolonged gastrointestinal residence time and controlled release might make contribution to H. Pylori clearance12.  Ethyl cellulose microspheres containing ciprofloxacin were prepared and evaluated for in-vitro performance of ciprofloxacin. It was concluded that ethyl cellulose microspheres showed reproducible results, with good mucoadhesive properties and good surface morphology13.  The microspheres of carprofen prepared by emulsion solvent evaporation method. It was seen that, drug: polymer ratio has an impact on the drug encapsulation efficiency and in vitro release14.  Glipizide mucoadhesive microspheres containing chitosan was prepared by simple emulsification phase separation technique using glutaraldehyde as a cross linking agent. The in vitro Studies showed that mucoadhesive microspheres of glipizide could sustain the release of the drug for more than 12 hours. In vivo Study was demonstrated on Albino Wistar rats and they observed the significant hypoglycemic activity of the mucoadhesive microsphere of glipizide15.  Mucoadhesive microspheres of atenolol and propranolol were prepared by an interpolymer complexation poly (acrylic acid) (PAA) with poly (vinyl pyrrolidone) (PVP) to increase gastric residence time by solvent diffusion method. They observed that release rate of atenolol from the complex microspheres was

16 slower than the poly vinyl pyrrolidone microspheres at pH 2.0 - 6.8 .

6.3. Objectives of the study: The specific objective of the study is to formulate and evaluate mucoadhesive microsphere of aceclofenac for oral drug delivery with an aim to:

 Achieve controlled release of the drug  Site specific drug delivery  Better therapeutic efficacy by local drug delivery  Increase the stability of drug  Avoid gastro intestinal disturbances associated with conventional anti-inflammatory therapy. b) MATERIALS AND METHODS: 7.1. Source of data: Preliminary data required for the experimental study would be obtained from, 1. Krupanidhi College of Pharmacy Library 2. Scientific Journals and Articles - International Journal of Pharmaceutical Sciences - International Journal of Pharmaceutics - Saudi Pharmaceutical journals - Advanced Drug Delivery Reviews 3. WHO Site and FDA Guideline website 4. Science direct and Helinet-RGUHS web sites.

7.2. Method of collection of data (including sampling procedure, if any): The data which is related to formulation and method development would be collected from various standard books, journals, patents, WHO and FDA guidelines and other sources like computer database.

The steps involved in the methodology would be: 1. Selection of polymers with mucoadhesive property. 2. Preformulation studies of the drug and in combination with excipients. 3. Selection of suitable technique such as emulsification-solvent evaporation, ionic gelation etc. 4. Preparation of mucoadhesive microspheres by selected technique. 5. Evaluation of mucoadhesive microspheres for  Drug content  Drug entrapment efficiency  Particle size & size distribution  SEM studies  Swelling index  Ex-vivo mucoadhesion test  In- vitro drug release studies.  Permeation studies  Short term stability studies 7.3. Does these studies require any investigation or intervention to be conducted on patients or other humans or animals? If so, please describe briefly. Yes, ex-vivo bioadhesion studies and permeation studies would be carried out on porcine gastric mucosa as the model membrane. The mucosal membrane would be collected, processed by removing unwanted connective and adipose tissues and equilibrated with phosphate buffer and would be studied for the above-mentioned tests. 7.4. Has ethical clearance been obtained from your institution in case of 7.4? Applied for c) LIST OF REFERENCES: 1. Sinha VR, Singla AK, Wadhawan S, Kaushik R, Kumria R. Chitosan microspheres as a potential carrier for drugs. Int J Pharm. 2004; 274: 1–33. 2. Alagusundaram M, Madhu Sudana Chetty C, Umashankari K Attuluri Venkata Badarinath, Lavanya C and Ramkanth S Microspheres as a novel drug delivery system. Int J Chem Tech Res. 2009:1(3):526-34. 3. Chowdary KPR, Rao YS. Mucoadhesive microspheres for controlled drug delivery. Bio pharm bulletin.2004; 27 (11): 1717-24. 4. Harshad Parmar, Sunil Bakliwal, Nayan Gujarathi, Bhushan Rane, Sunil Pawar. Different methods of formulation and evaluation of mucoadhesive microsphere. Int J App Bio Pharm Tech.2010;1(3):1157-66. 5. Simina Dreve , Iren Kacso , Adriana Popa , Oana Raita , Felicia Dragan , Bende A, Borod Gh, Bratu I. Structural investigation of chitosan-based microspheres with some anti-inflammatory drugs. J Mol Str. 2011; 997 :78–86 6. Tuncay M, Calis S, Kas HS, Ercan MT, Peksoy I, Hincal AA. In vitro and in vivo evaluation of diclofenac sodium loaded albumin microspheres. J Microencapsul., 2000;17 (2): 145-55, 7. Tomoko Oosegi, Hiraku Onishi, Yoshiharu Machida. Novel preparation of enteric-coated chitosan- prednisolone conjugate microspheres and in vitro evaluation of their potential as a colonic delivery

system. Euro J Pharm Biopharm. 2008; 68:260–6. 8. Haznedar.B, Dortun.B. Preparation and in vitro evaluation of Eudragit microspheres containing acetazolamide. Int J Pharm. 2004;269 :131–40. 9. Yunying Tao, Yifan Lu, Yinjing Sun, Bing Gu, Weiyue Lu, Jun Pan. Development of mucoadhesive microspheres of acyclovir with enhanced bioavailability. Int J Pharm.2009; 378;30–6. 10. Martinac.A, Filipovi´c-Grˇcic.J, Voinovich.D, Perissutti.B, Franceschinis.E, Development and bioadhesive properties of chitosan-ethylcellulose microspheres for nasal delivery. Int J Pharm.2005; 291: 69–77. 11. Arya RKK, SinghR, Vijay Juyal. Mucoadhesive microspheres of famotidine: preparation Characterization and in vitro Evaluation. Int J Eng Sci Tech. 2010; 2(6): 1575-80. 12. Yellanki SK, SinghJ, Syed JA, RajkamalBigala, SharadaGoranti, Nerella NK. Design and Characterization of Amoxicillin trihydrate Mucoadhesive Microspheres for Prolonged Gastric retention. Int J Pharm Sci and Drug Res 2010; 2(2): 112-114. 13. Hardenia SS, Jain A, Patel R, AnuKaushal. Formulation and Evaluation of Mucoadhesive Microspheres of Ciprofloxacin. J Adv Pharm Edu Res. 2011; 1(4): 214-24. 14. Sathiya Sundar R,Murugesan A,Venkatesan P,Manavalan R.Formulation development and evaluation of Carprofen Microspheres.2010;2(3):1674-76. 15. Patel JK, Patel RP, Amin AF, Patel MM. Formulation and evaluation of mucoadhesive glipizide microspheres AAPS pharm Sci Tech 2004 Oct 12.avalable from URL: http://www.aapspharmscitech.org. 16. Prajapati SK, tripati P, Ubaidulla U, Anand V. Design and Development of Gliclazide Mucoadhesive Microcapsules: in vitro and in vivo Evaluation. AAPS Pharm Sci Tech.2008 March; 9(1).available from URL:http://www.aapspharmsciech.org