Preparation of non infringing strategy: Flow Chart Prior art search

Transcription

1 2. MATERIALS & METHODS 2.1 Preparation of Non-Infringement opinion Selection of a product to work on. Product marketing team, International Export team, experts and Buyers provided the name of Product which was required for export and marketing there according to the demand of the drug product in prevalent disease condition and profitability in that geographical area. Preparation of non infringing strategy: Flow Chart Prior art search Evaluation of patent identified as relevant Ideas against claims, strategies, objectives, and resources Patented and develop product concepts proposed plan for a proper strategy Analysis of profit (Costs, Market, & Competition, etc) Technical and commercial product development Implementation of Proposed product Commercialization Compilation of all the details of oral capsule formulation comprising Tianeptine sodium. Search for innovator s formulation The capsule formulation was not available in public domain search. Designing of the formulation A formulation was designed after thoroughly studying the various factors of compatibility study and various literature studies supported by laboratory Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 18 of 124

2 experimentation work. The strengths of inactive ingredients were calculated on the behalf of usual ranges of inactive ingredients prescribed in the text books and formularies Retrieve detail information related to patents existing in each of these countries for Tianeptine sodium Capsules Formulation of a search strategy A search strategy was formulated after understanding the client s proposed product, process and technology thoroughly. Key search terms were decided for doing searches on available electronic databases like etc Determine Key Search Areas Key search areas were determined viz. product patents, process patents, formulation patents, polymorph patents indication and method of use patents Perform searches Searches were conducted on available electronic databases with each of the predetermined key terms and results of the search were downloaded. The searching for patents available in these countries was performed on electronic databases available for these countries Patent Search Strategy It was done to save time & it allowed us to search information at different places & to find a larger amount of relevant information Basis of the search strategy The search strategy was formulated after understanding the client s proposed product, process and technology thoroughly. Following key conclusions were drawn after carefully examining the proposed project: The client wanted to manufacture and sell a pharmaceutical formulation in the countries viz. US, U.K and Russia. The proposed formulation was Capsule and comprised of Tianeptine sodium. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 19 of 124

3 The proposed process of manufacturing was conventional wet granulation method. The product is indicated for the treatment of depression Available Electronic Databases There are many electronic databases available on internet. Some of the most reliable electronic databases useful for the proposed project were: Orange Book: Approved Drug Products with Therapeutic Equivalence Evaluations United States Patent and Trade Mark Office (USPTO) European Patent Office (EPO): Espacenet Patent search The World Intellectual Property Organization (WIPO) Key search Areas Key search areas were identified to find out relevant patents. Patents on Compositions/formulations comprising of Tianeptine Product, Process and Polymorph patents of API Patent search with Assignee Names for patents on Process/ Technology for making the formulation i.e. Capsules Search terms for patent searching Following search terms were used to perform searching on the selected databases: I) For patents on tablet formulation comprising of Tianeptine II) For patents of Active Pharmaceutical Ingredients (APIs) For product/ process patents Patent number search for any prior art patent cited in the downloaded relevant patents and/or Manual search in Merck Index and/or For polymorph patents Tianeptine AND Polymorph AND Crystal AND Hydrate AND Solvate AND Amorphous For patent search with assignee names (For process/ technology for making the formulation i.e. Capsules) Teva Pharma AND Capsules AND Dosage form Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 20 of 124

4 2.1.5 Prepararation of a detailed Non-infringement opinion for all these countries with respect to patents existing in each of these countries for Tianeptine sodium Capsules Identification of relevant patents Out of the all downloaded patents, the relevant patents were identified Infringement analysis For each of the identified relevant patents infringement analysis was done by following the step procedure viz. Claim Construction and Claim Comparison Conclusion Based on the results of infringement analysis conclusions were drawn Designing of a Formulation Consisting Of Tianeptine Description of the Proposed Formulation Proposed Formulation Following formulation was designed. This formulation is proposed to be sold in US, U.K. and Russia Dosage Form: Capsule Route of Administration: Oral Table 1: Formula of the Proposed Formulation B. Size: 10,000 Capsules Table 2.1 Batch formula of BRUTINE (Tianeptine sodium Capsules 12.5 mg) for Capsules. Ingredients Spec Standard Quantity (Kg) Quantity required (Kg) Quantity per Capsules (mg) Function A: Granules Preparation Tianeptine sodium BP * * Active Microcrystalline cellulose (PH 102) BP Diluent Microcrystalline cellulose Plain BP Diluent Starch BP Diluent Magnesium stearate BP Lubricant Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 21 of 124

5 Ingredients Spec Standard Quantity (Kg) Quantity required (Kg) Quantity per Capsules (mg) Function Colloidal anhydrous silica BP Glidant Purified Talc BP Glidant Croscarmellose sodium BP Disintegrant Sodium starch glycolate BP Disintegrant Crospovidone BP Disintegrant B: Capsules shell: Empty Hard gelatin capsule shells Light green-light green; Size 5 IHS 10200nos 10200nos 1 no Protective shell * Overages of Tianeptine sodium- 5% Average weight of Capsule is mg Note: The quantities of inactive ingredients mentioned in the formula are based on the ranges of inactive ingredients prescribed in text books. These were required to be optimized experimentally to get the final formula. Indications The product is indicated for the treatment of depression Capsule Manufacturing Process Capsule was prepared by the conventional wet granulation for granulation compounding method. Major steps involved in the manufacturing process were depicted here. Dry Mixing of Tianeptine with microcrystalline cellulose Granulation of mixture with water in a high shear granulator Wet mass was screened,dried and milled Milled granules were blended with Crospovidone and Magnesium Stearate to produce the compression mix & add the other Excipients Granules were compressed at size Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 22 of 124

6 Granules were filled in the capsules Note: The above mentioned process is based on typical wet granulation manufacturing processes prescribed in text books. This process was required to be optimized experimentally. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 23 of 124

7 2.2 Preparation of Dossier in EU-CTD format for BRUTINE (Tianeptine Sodium Capsules 12.5 mg) for U.K. The Common Technical Document (CTD) is provides five modules. (Notice to Applicants, Volume 2B, 2006) Requirement for module 1 Requirement for module 2 Requirement for module 3 Requirement for module MODULE 1 Administrative Information and prescribing information Refer to EU CTD MODULE 1 inannexure MODULE 2 Common Technical Document Summaries Refer to 3.RESULT & DISCUSSION & 4.SUMMARY AND CONCLUSION MODULE 3 QUALITY Table of contents of Module 3 Refer to Comprehensive Table of Contents in Annexure DRUG SUBSTANCE Refer to DMF (Open Part) in Annexure DRUG PRODUCT Description and Composition of the Drug Product Nomenclature INN: Tianeptine Sodium USAN: Tianeptine Sodium Molecular formula: C 21 H 24 ClN 2 NaO 4 S CAS: ATC code: N06AX14 Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 24 of 124

8 Chemical Name: 7 - [ ( 3 Chloro 6, 11 dihydro 6 methyldibenzo [c, f] [1, 2] thiazepin 11 yl) - amino] heptanoic acid S, S - dioxide Chemical Structure: Molecular Weight: Tianeptine is a tricyclic antidepressant. Available in market as a racemate & its more active form is l-isomer. The primary metabolites have the same basic structure; There are -2 & 4 -carbons respectively on side chain. Table 2.2 Product Information of Tianeptine sodium Capsules 12.5 mg Product Name BRUTINE (Tianeptine sodium Capsules 12.5 mg) Product Code R/018 Description Light green-light green Hard gelatin Capsules, Size 5. Label Claim Each Capsules contains : Tianeptine Sodium BP 12.5 mg Average Wt mg Dosages form Hard Gelatin Capsules Shelf Life 24 Months Storage Condition Store at temperature not exceeding 25 0 C, Keep in dry place away from light. Standard Batch Size Capsules Batch No. BC B. Size: 1,00,000 Capsules Table 2.3 Batch formula of BRUTINE (Tianeptine sodium Capsules 12.5 mg) for capsules Ingredients Spec Standard Quantity (Kg) Quantity required (Kg) Quantity per Capsules (mg) Function A: Granules Preparation Tianeptine sodium BP * * Active Microcrystalline cellulose (PH 102) BP Diluent Microcrystalline cellulose Plain BP Diluent Starch BP Diluent Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 25 of 124

9 Ingredients Spec Standard Quantity (Kg) Quantity required (Kg) Quantity per Capsules (mg) Function Magnesium stearate BP Lubricant Colloidal anhydrous silica BP Glidant Purified Talc BP Glidant Croscarmellose sodium BP Disintegrant Sodium starch glycolate BP Disintegrant Crospovidone BP Disintegrant B: Capsules shell: Empty Hard gelatin capsule shells Light green-light green; Size 5 IHS nos nos 1 no Protective shell Average weight of Capsule is mg * Overages = 5% Primary Packing Components Printed Aluminium foil PVC Secondary Packing Components: Carton Tertiary Packing Components The cartons are to be packed in a corrugated fiberboard box depending on the type of packing details. All the cartons were poly-laminated in a group of 10 cartons, which protects the cartons from humidity and other damages Pharmaceutical Development Components of the Drug Product a) Drug Substance Specifications of Drug Substance Material Name Tianeptine sodium BP Table 2.4 Specification of Tianeptine sodium BP Test parameters Specification Reference Appearance White to yellowish powder Very hygroscopic powder B.P. Solubility Freely soluble in water and methanol B.P. Identification Reaction positive B.P. Related substance Individual impurity: Not more than 0.1% B.P. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 26 of 124

10 Total impurities: Not more than 0.4% B.P. Methanol Not more than 0.3% B.P. Dichloromethane Not more than 0.06% B.P. Ethyl acetate Not more than 0.5% B.P. Water Not more than 5.0% B.P. Assay (on anhydrous basis) 99%-101.0% B.P. Testing procedure: Tianeptine sodium BP Identification A. Infrared absorption spectrophotometry. Comparison: Comparision European Pharmacopoeial reference spectrum of Tianeptine Sodium B.P sodium. B. It gives reaction (a) of sodium. Assay g dissolved in 50 ml of anhydrous acetic acid R. titrated with 0.1 M perchloric acid, End-point were determined by potentiometric titration. 1 ml of 0.1 M perchloric acid is to mg of C 21 H 24 ClN 2 NaO 4 S. b) Excipients Table 2.5 Function of Excipients Excipients Specification Function Microcrystalline cellulose (PH 102) BP Diluent Microcrystalline cellulose Plain BP Diluent Maize Stach BP Diluent Magnesium stearate BP Lubricant Colloidal anhydrous silica BP Glidant Purified Talc BP Glidant Croscarmellose sodium BP Disintegrant Sodium starch glycolate BP Disintegrant Crospovidone BP Disintegrant Capsules shell: Empty Hard gelatin capsule shells Light green/light green; Size 5 IHS Protective shell Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 27 of 124

11 Drug Product Formulation Development FORMULATION DEVELOPMENT A). Scope: This development report is applicable to development of Tianeptine sodium Capsules 12.5 mg using the facility of Brawn Laboratories Ltd., located at Faridabad, India. B). Systematic Approach: A Systematic approach to Tianeptine sodium Capsules 12.5 mg was designed and product development activities were sub divided into literature search, formulation development and process development. The various studies carried out during these phases were given below. C). Literature Search: Preliminary activities in a drug product development project started with a literature review & prior art search in reference books on the pharmaceutical and analytical parameters. Reference books, the current United State Pharmacopoeia, British pharmacopoeia, European Pharmacopoeia, Physician s Desk Reference, World intellectual property Organization, Martindale: The complete drug reference and Merck Index were reviewed on physical and chemical parameters on the active ingredient and its formulations. An extensive online internet searching relating to drug substance and the drug product was done. Field of the Research: The present research is related to development of a novel pharmaceutical formulation - Tianeptine sodium Capsules 12.5 mg D). Background of the Research i) Drug Information: Tianeptine (Stablon, Coaxil, Tatinol) (Tianeptine, 2011) is a drug product useful for treating major depressive cases like (mild, moderate, or severe). It is structurally similar to tricyclic antidepressants, but having different pharmacological properties. Actions were also found on the NMDA and AMPA receptors. Recently, Tianeptine was approved in France, there it is marketed & is manufactured by Laboratoires Servier SA; & is also marketed by a other EU countries under brand Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 28 of 124

12 name of Coaxil also in Latin America & Asia as "Tatinol" & Stablon" and but it is not available in US or UK. E) Active Sourcing, Evaluation & Purchasing Sourcing for Active Raw Material: We selected International Suppliers US, EU, Asian Potential Suppliers List: Request samples and COA and Specifications. We Selected & evaluated at least two suppliers source. 1. Vivan Life sciences, Thane, Mumbai (India) 2. Shanghai soyoung Biotech Inc. Refer to COA S in Annexure 2 F) Active Testing Tianeptine i) Drug information (Active ingredient Tianeptine Sodium) Physicochemical properties Chemical structure Chemical name: [ 3 chloro 6 - methyl - 5,5 dioxo - 6,11-dihydro - (c, f)- dibenzo - (thiazepine) yl) amino] -7 heptanoic acid, sodium salt. Molecular formula: C 21 H 24 ClN 2 NaO 4 S Molecular weight: Appearance: White or yellowish powder, very hygroscopic. Solubility: Freely soluble in water, methylene chloride & methanol. ii) Testing of Active Substance sample: Chemical testing by the R&D lab as per Pharmacopoeia method (as per Ph Eur & BP) &/ Inhouse. Protocol: BP 2011 was followed for the analysis. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 29 of 124

13 iii) Stability Summary and Conclusions Substance : Tianeptine Sodium B.P Proposed expiry Storage Stability studies : 24 months : Store < 25 C, Protect from light and moisture. :Accelerated stability study at 40 C & Long term stability study at 25 0 C are carried out and stability data are attached. iv) Packing Condition Tianeptine Sodium B.P is packaged in a light resistant cardboard container Package 10 kg/drum, or according to customers. For detail refer to DMF (Open Part) in Annexure 2. G) Excipients Evaluation of formulation with suitable excipients: Excipient compatibility tested using DSC method and stability assessment was done. Table 2.5 Function of Excipients Excipients Specification Function Microcrystalline cellulose (PH 102) BP Diluent Microcrystalline cellulose Plain BP Diluent Maize Stach BP Diluent Magnesium stearate BP Lubricant Colloidal anhydrous silica BP Glidant Purified Talc BP Glidant Croscarmellose sodium BP Disintegrant Sodium starch glycolate BP Disintegrant Crospovidone BP Disintegrant Capsules shell: Empty Hard gelatin capsule shells Light green/light green; Size 5 IHS Protective shell All excipients were compatible with each other & with API H). Container Closure System Primary Packing Components Printed Aluminium foil Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 30 of 124

14 PVC Secondary Packing Components 1. Carton Tertiary Packing Components The cartons were packed in a corrugated fiberboard box depending on the type of packing details. All the cartons are poly-laminated in a group of 10 cartons, by which cartons were protected from humidity and other damages. I) R & D BATCHES: Optimization of Product (Tianeptine sodium Capsule 12.5 mg) Table 2.6 Composition of Tianeptine sodium Capsule 12.5 mg formulations S. No Ingredients F1 F2 F3 F4 F5 F6 F7 F8 1 Tianeptine sodium A : Excipients 2 Starch Magnesium stearate Colloidal anhydrous silica Purified Talc Croscarmellose sodium Sodium starch glycolate Crospovidone B: Capsules shell: Empty Hard gelatin capsule shells Light green-light green; Size 5 1 no Batches F4, F5, & F6 were selected for further study & optimized for the further study because these provide better dissolution profile. Development of the formulation here was mainly based on the type of Excipients in different combinations were used so as to get desire in vitro drug release from Capsule. Tianeptine sodium is water soluble drug, and dose taken is 12.5 mg. So, in the present study attempts were made to get capsule with desire drug release profile of the capsule. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 31 of 124

15 Manufacture of capsule As described in the methodology chapter the, Granules prepared by wet granulation method by mixing the ingredients. J) SCALE-UP BATCHES: In scale-up 3 batches were selected & prepared out of which one Batch was found to provide appropriate & better results, which was selected for the proposed formulation as Pilot Scale Batch. i) Formula of the Proposed Formulation B. Size: 10,000 Capsules Table 2.1 Batch formula of BRUTINE (Tianeptine sodium Capsules 12.5 mg) for 10,000 Capsules Ingredients Spec Standard Quantity (Kg) Quantity required (Kg) Quantity per Capsules (mg) Function A: Granules Preparation Tianeptine sodium BP * * Active Microcrystalline cellulose (PH 102) BP Diluent Microcrystalline cellulose Plain BP Diluent Maize Starch BP Diluent Magnesium stearate BP Lubricant Colloidal anhydrous silica BP Glidant Purified Talc BP Glidant Croscarmellose sodium BP Disintegrant Sodium starch glycolate BP Disintegrant Crospovidone BP Disintegrant B: Capsules shell: Empty Hard gelatin capsule shells Light green-light green; Size 5 IHS 10200nos 10200nos 1 no Protective shell Average weight of a Capsule is mg Overages = 5% Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 32 of 124

16 ii) Manufacturing Process and process Controls PROCESS FLOW DIAGRAM FLOW CHART OF MANUFACTURING Approved Raw Material & Excipients Sifting Blending Granulation Capsule Filling Capsule Polishing/Dedusting Final Packing Transfer to Finished Goods Store Dispatch Manufacturing process a) Weighing and Checking Weights were checked for the raw materials as per the formulation sheet (i.e. page No.3) The AR No. of all the materials was also checked as per the issuance sheet. b) Sieving: Tianeptine Sodium was sieved through #80 and others through # 60 sieve fitted to a Vibrosifter and collected in separate Intermediate Process Containers c) Granulation: Granulation was done with the help of Purified water with the following Raw material in quantities as mentioned herewith. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 33 of 124

17 Ingredients Standard Specification Quantity(Kg) Tianeptine sodium * BP Microcrystalline cellulose (PH 102) BP Microcrystalline cellulose Plain BP Maize starch BP Magnesium stearate BP Colloidal anhydrous silica BP Purified Talc BP Croscarmellose sodium BP Sodium starch glycolate BP Crospovidone BP d) Blending (Lubrication): All the sieved materials were sifted in Double cone Blender and blended for 30 minutes at 10 RPM. Unloaded the granules in double polythene lined drums and closed tightly. Labeled the drums appropriately and recorded all the details including Product Name, Batch No., Batch size, Mfg. Date. Request slip was sent to QA department to draw the sample for bulk analysis done by QC department. Ensured that temperature and humidity in the Capsule filling area is 25 C± 2 C, & RH: NMT 45 %± 5% respectively. Undue storage of granules Avoided. Granules were taken up immediately for filling into Capsule shells. e) In-Process Checking In process parameters at regular intervals were monitored and recorded in the following formats. Q.A. / Q.C. for to do in-process checks as per the norms set by them. Send the sample of blend to QC for Assay. Attached the requisition slip and QC In-process results slip received from QC. After blend was released from QC, bulk was filled into capsules as per specification. Storage Condition :Store below 25 º C, Protect from light TEST LIMIT Description Off white granular Powder Assay : Tianeptine sodium BP 98.0 to 102.0% Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 34 of 124

18 Moisture content NMT 2 % f) Quality Assurance / Quality Control Approval After getting Quality Approval, the Blend was transfered to Capsule Filling dept. and the product was filled as per specifications and instructions g) Filling Ensured that temperature and humidity in the filling area was 25 C± 2 C, NMT 45%± 5% respectively. Undue storage of granules Avoided. Granules were taken up immediately for filling into Capsule shells. The Capsule Filling Machine was set as per respective SOP. In process parameters at regular intervals were monitored and recorded in the following formats. Q.A. / Q.C. for to do in-process checks as per the norms set by them. The filling parameters were adjusted as per the standards given below:- Table 2.7 Capsule Filling parameters as per the standards Parameters Standards Appearance Light green / Light green hard gelatin capsule size 5, containing off white granular powder. Standard Fill Weight of Capsule 93mg + 2 % Weight of Filled Capsule mg + 2 % Weight of 20 Capsules 2.41 g Individual Weight Variation + 10 % Disintegration Time NMT 20 minutes Locking Length of Filled Capsule 10 ± 0.2 mm h) In-Process Checking Frequency In process parameters at regular intervals were monitored and recorded in the following formats. Q.A. / Q.C. for to do in-process checks as per the norms set by them Weight of 20 Capsules Fill Variation Disintegration time Locking Length of Filled Capsule i) Rejection Every 30 minutes Every 120 minutes Every 120 minutes Every 120 minutes Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 35 of 124

19 The Rejection/Recovery of the Blend/Capsules were collected, recorded, labeled appropriately and destroyed as per respective SOP j) Quality Assurance / Quality Control Approval Request slip to Q.A. Department was sent to draw the sample for finished product analysis to be done by QC Dept. After getting Quality Approval, Capsules were transfered the to packing dept. and packed the product as per current packaging specifications and instructions k) Packaging: Batch Issuance Slip from Manufacturing to Packing Table 2.8 Requisition of Packing Material Item code Items UOM Standard quantity PRIMARY PACKING COMPONENTS B.P.F769 Aluminum Blister foil Size 185mm Kg B.P.01 8 PVC Film Size 188 mm clear Kg SECONDARY PACKING COMPONENTS B.P.A A26 Printed cartons Tianeptine sodium Capsules 10 x 10 Nos 1000 B.P.J0 11 Corrugated Box G-50 Nos B.P.T7 6 Literature Tianeptine sodium Capsules Nos 1000 B.P.K 10 Plastic film 500 mm (20 ) Kg l) Batch Details Instructions and Certification Transfer the batch IPCs to the packing cubicle and check if the number of containers received for the batch. Load the IPC in machine hopper in serial order Load the Printed foil on winding shaft of the Blister Packing machine Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 36 of 124

20 Perform the Leak test on the samples drawn during machine setting as per the relevant SOP. Start the machine only after the Blister passes the leak test and record the observation. Set the sealing heater temperature. Monitor it initially and there after every 60 minutes by Production Department and after every 120 minutes by QA Department. Record the observations in the In Process Sheet. iii) Testing Procedure: Average weight: 20 Capsules were randomly selected & then average weight was calculated by following equation: Average Wt. = Wt. of 20 Capsules Average weight of Capsules = mg ±10% Uniformity of weight: 20 Capsules were weighed and then average weight of 20 capsules was calculated. Then 20 Capsules were weighed individually. The Capsules having highest weight and the Capsules having lowest weight in the above 20 Capsules were find out by:. Calculate the maximum & minimum deviation as follows: (WH -A) x 100 Maximum deviation (+ve) = A (A - WL) x 100 Minimum deviation (-ve) = A Where- A: Average weight of Capsules WH: Highest weight of Capsules WL: Lowest weight of Capsules Limits: ±10% Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 37 of 124

21 Moisture content (In process Control Test for Granules) Moisture content of the granules was determinated with infrared dryer (Sartorius MA 100), by evaporation of bound and unbound moisture. Approximately 4 g of granules was tested immediately on the manufacturing floor.mass change was determined and water content was expressed as percentage of the total mass. Sample was heated until C. Measurement was terminated when weigh loss in percent is less than 0.1 % in 50 seconds. Moisture content: (S1 S2) X 100 S1= Initial sample; S2 = Sample after heating Limit: NMT 2% Disintegration time: Placed 1 Capsule each in 6 tubes of the basket with a disc. The apparatus was operated using the medium as specified in Pharmacopoeia, the apparatus was maintained at 37 ± 2 C, as the immersion fluid. At the end of the mentioned time, the basket was lifted and observed the capsules. All of the capsules were disintegrated completely. Limits: NMT 30 minutes. Dissolution: Medium:- 0.1 N Hcl ; 900 ml. Apparatus:- 2: 50 rotation per minute Time:- 45 minutes. Procedure Apparatus II was used. The medium 900 ml of 0.1M hydrochloric acid was used and the paddle was rotated at 50 revolutions per minute. A sample of 10 ml was withdrawn from the medium and filtered. The absorbance of the filtered sample was measured, diluted if necessary with 0.1M Hcl, at the maximum at 220 nm, using 0.1M hydrochloric acid in the reference cell, and compared with that of a standard solution in 0.1M hydrochloric acid having a known concentration of Tianeptine sodium equivalent to about 10 to 20 µg of C 21 H 24 ClN 2 NaO 4 S per ml. The total content of Tianeptine sodium in the medium was calculated. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 38 of 124

22 Analytical Procedure: Method of Analysis Details of Tianeptine on HPLC 1. Column:- C Mobile Phase: - Mobile phase consist of acetonitrile: 50 mm potassium dihydrogen phosphate, ph 3.5 (33:67, v/v/). 3. Flow rate: - It having pump at a flow rate of -1ml/min. 4. λ max: - Operating at -214 nm. Standard stock solutions Standard stock solutions of 100 µg/ml was prepared in mobile phase by dissolving the pure drug in it. Preparation of the calibration curves Aliquots were prepared by serial dilution method sufficient to produce ng/ml. Each solution was injected one by one and chromatogram was recorded at 214 nm. The peak area of drug was noted and calibration curve was plotted as peak area against concentration of drug. Analysis of capsule formulation The contents of twenty capsules were mixed and weighed accurately. Powder equivalent to specification transferred into a 100 ml volumetric flask, dissolved in water and sonicated for 5 min., the volume was made up with water, shaken well for 5 min. and then filtered, further absorbance value was noted in triplicate at 214 nm against reagent blank. Instrumental Conditions: Instrument: UV-VIS Spectrophotometer (Shimadzu make Model-1800) Wavelength: 214 nm Procedure The absorbance of the test solution was measured by using the UV-VIS Spectrophotometer at the maximum at 214 nm and record the spectra was recorded. Calculation: The content was calculated taking 562 as the value of A (1%, 1 cm) at the maximum at 214 nm. Test Abs potency = x x x x x Test wt Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 39 of 124

23 Microbiological Attributes Standard Operating Procedure is applicable for Microbial limit test in microbiology laboratory in Quality Control department at Brawn Laboratories Limited, Faridabad. 1. MICROBIAL CONTAMINATION: TOTAL BACTERIAL COUNTS TOTAL FUNGAL COUNTS OTHER MICROORGANISM: NMT 1000 CFU/G NMT 100 CFU/G ABSENT Refer to SOP- Microbial Limit Test in Annexure 2. Stability study: The objective of the study on BRUTINE (Tianeptine sodium Capsules 12.5 mg) was to find out the stability profile of capsules stored under accelerated stability study conditions. The Capsules were put in an inverted position so that there was a contact with the inner container. The batches kept under stability study were provided in the following table: Table 2.9 Batches used under stability study during F & D Dosage Batch No Manufacturing Scale Total Batch API Batch No. Date Site Size 12.5mg/ Capsule BC /12/2010 Faridabad Pilot scale Capsules BP/BPA_ 001/D/10/ mg/ Capsule BC /12/2010 Faridabad Pilot scale Capsules BP/BPA_ 001/D/10/ mg/ Capsule BC /12/2010 Faridabad Pilot scale Capsules BP/BPA_ 001/D/10/014 Finally the Commercial or Production scale batch was prepared after the better results obtained during course of Testing & Stability study performed. Summary: The report shows the stability data on BRUTINE (Tianeptine sodium Capsules 12.5 mg) Samples were stored for 6 months in the primary container closure system. Any storage-related changes occurring in the finished product were checked by means of control tests specified in stability study. The test was designed on basis of stability profile of Tianeptine sodium BP for the certain specific requirements of Capsules. Chemical stability does not affected by Storage of Capsules under accelerated Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 40 of 124

24 conditions for 6 months. There was not found any significant change in content value of BRUTINE (Tianeptine sodium Capsules 12.5 mg) compared to its initial value. Conclusion: The stability data demonstrated that all lots of the product BRUTINE (Tianeptine sodium Capsules 12.5 mg) remained within specifications at all times during the accelerated stability conditions at 40 C ± 2 C, 75% ± 5 % Relative humidity. Based on the above, we concluded that the product BRUTINE (Tianeptine sodium Capsules 12.5 mg) was stable for 6 months. The method used to conduct the stability studies was taken as ICH guidance document Q1A-(R2). The above study was performed at Brawn Laboratories Limited under the supervision of our analyst manager. iv) Batch formula Production; Batch no. BC Size: 1, 00,000 Capsules Table 2.3 Batch formula of BRUTINE (Tianeptine sodium Capsules 12.5 mg) for capsules Ingredients Spec Standard Quantity (Kg) Quantity required (Kg) Quantity per Capsules (mg) Function A: Granules Preparation Tianeptine sodium BP * * Active Microcrystalline cellulose BP Diluent (PH 102) Microcrystalline cellulose Plain BP Diluent Maize Starch BP Diluent Magnesium stearate BP Lubricant Colloidal anhydrous silica BP Glidant Purified Talc BP Glidant Croscarmellose sodium BP Disintegrant Sodium starch glycolate BP Disintegrant Crospovidone BP Disintegrant B: Capsules shell: Empty Hard gelatin capsule shells Light green-light green; Size 5 IHS nos Average weight of Capsule is mg * Overages = 5% nos 1 no Protective shell Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 41 of 124

25 Overages 5 %, Overages taken due to waste of raw materials that occured during manufacturing of finished products v) Critical parameters: Table 2.10 Critical Parameters & Acceptance criteria STAGE PARAMETER ACCEPTANCE CRITERIA Blending Assay 98.0 to 102.0% Moisture Content NMT 5 % Capsule Filling Average fill 93 mg ± 2% Fill Variation ± 10 % Disintegration time NMT 20 minutes Length 5 ±0.2 mm Chemical Analysis As per Finished Product Specification Microbial Analysis Total bacteria count (NMT 1000 cfu/gram). Total fungal count (NMT100 cfu/gram) Hold Time Study Blend Assay 98.0 to 102.0% Moisture Content NMT 5 % Microbial Analysis Total bacteria count (NMT 1000 cfu/gram). Total fungal count (NMT100 cfu/gram) Capsule Filling Average fill 93 mg ± 2% Fill Variation ± 10 % Disintegration time NMT 20 minutes Length 5 ±0.2 mm Chemical Analysis As per Finished Product Specification Microbial Analysis Total bacteria count (NMT 1000 cfu/gram). Total fungal count (NMT100 cfu/gram) Blister Packing Leak Test None of the strips shall fail in the leak test Acceptance Criteria i) All the tests performed during the process should qualify the specifications. ii) Any deviation from the acceptance criteria of the specific check point shall be Reported and decision should be taken for the acceptance, rejection, change in the Formulation /process Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 42 of 124

26 vi) Batches used for Validation Table 2.11 Batches used for validation during F & D Batch No Batch Size Mfg Date Exp Date BC Capsules 06/12/ /11/2012 Batch No Batch Size Mfg Date Exp Date BC Capsules 10/12/ /11/2012 Batch No Batch Size Mfg Date Exp Date BT Capsules 11/12/ /11/2012 vii) Batch Analyses: Refer to Batch Analyses in 3. RESULT & DISCUSSION viii) Dissolution Profile Comparison by using Model Independent Approach Using a Similarity Factor a) Samples details: Sample: BRUTINE (Tianeptine sodium Capsules 12.5 mg) Manufactured By: BRAWN LABORATORIES LTD Batch No. : Batch no. BC Manufacturing Date: 12/2010 Expiry Date: 11/ 2012 Description: Light green-light green Hard gelatin Capsules, Size 5.Containing off white granular powder. Average Weight: mg b) Samples Details of Innovator s Product: Sample: STABLON (Tianeptine sodium Tablets 12.5 mg) Manufactured By: Les Laboratories Servier, (France) Batch No. : LSC0502 Manufacturing Date: 11/2010 Expiry Date: 10/ 2012 Description: Sugar coated Tablets. Average Weight: 180 mg Chemicals: Hydrochloric Acid Reagent: 0.1 N hydrochloric acid Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 43 of 124

27 c) Dissolution test procedure: Equipments: Tablet Dissolution Test Apparatus - USP 24 (Model No. TDT-06P, Electrolab, India); Instrument: Shimazdzu UV-1601 Medium: 0.1 N Hcl; 900 ml. Apparatus 2: 50 rpm Time: 45 minutes. Procedure Apparatus II was used. The medium 900 ml of 0.1M hydrochloric acid was used and the paddle was rotated at 50 revolutions per minute. A sample of 10 ml was withdrawn from the medium and filtered. The absorbance of the filtered sample was measured, diluted if necessary with 0.1M hydrochloric acid, at the maximum at 220 nm, using 0.1M hydrochloric acid in the reference cell, and compared with that of a standard solution in 0.1M hydrochloric acid having a known concentration of Tianeptine sodium equivalent to about 10 to 20 µg of C 21 H 24 ClN 2 NaO 4 S per ml. The total content of Tianeptine sodium in the medium was calculated. d) Comparitive Dissolution Profile of BRUTINE and STABLON Figure: 2.1 Comparative Dissolution Profiles of BRUTINE and STABLON e) Result Dissolution profile of BRUTINE was compared with STABLON & Factor F 2 was found to be 71%. On this basis BRUTINE was bioequivalent with STABLON. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 44 of 124

28 This also provided recommendations for dissolution tests to help ensured continuous drug product quality in each & every steps and performance after some post approval manufacturing changes Overages Refer to section Formulation Development Physiochemical and Biological Properties Refer to section Formulation Development Manufacturing Process Development Refer to section Formulation Development Container Closure System Refer to section Container Closure System Microbiological Attributes Standard Operating Procedure is applicable for Microbial limit test in microbiology laboratory in Quality Control department at Brawn Laboratories Limited, Faridabad. MICROBIAL CONTAMINATION: TOTAL BACTERIAL COUNTS NMT 1000 CFU/G 1. TOTAL FUNGAL COUNTS NMT 100 CFU/G OTHER MICROORGANISM: ABSENT Refer to SOP- Procedure for Microbial Limit Test in Annexure Compatibility Not applicable Manufacture Manufacturer Brawn Laboratories Ltd. 13, N.I.T. Industrial Area, Faridabad , (Haryana), India Telephone: , , Fax: Website: Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 45 of 124

29 Batch Formula Table 2.2 Product Information of Tianeptine sodium Capsules 12.5 mg Product Name BRUTINE (Tianeptine sodium Capsules 12.5 mg) Product Code R/018 Description Light green-light green Hard gelatin Capsules, Size 5. Label Claim Each Capsules contains : Tianeptine Sodium BP 12.5 mg Average Wt mg Dosages form Hard Gelatin Capsules Shelf Life 24 Months Storage Condition Store at temperature not exceeding 25 0 C, Keep in dry place away from light. Standard Batch Size Capsules Batch No. BC B. Size: 1,00,000 Capsules Table 2.3 Batch formula of BRUTINE (Tianeptine sodium Capsules 12.5 mg) for capsules Ingredients Spec Standard Quantity (Kg) Quantity required (Kg) Quantity per Capsules (mg) Function A: Granules Preparation Tianeptine sodium BP * * Active Microcrystalline cellulose BP Diluent (PH 102) Microcrystalline cellulose BP Diluent Plain Starch BP Diluent Magnesium stearate BP Lubricant Colloidal anhydrous silica BP Glidant Purified Talc BP Glidant Croscarmellose sodium BP Disintegrant Sodium starch glycolate BP Disintegrant Crospovidone BP Disintegrant B: Capsules shell: Empty Hard gelatin capsule shells Light green-light green; Size 5 IHS nos nos 1 no Protective shell Average weight of Capsule is mg * Overages = 5% Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 46 of 124

30 Description of Manufacturing Process and process Controls Process Flow Diagram Flow Chart of Manufacturing Approved Raw Material & Excipients Sifting Blending Granulation Capsule Filling Capsule Polishing/Dedusting Final Packing Transfer to Finished Goods Store Dispatch Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 47 of 124

31 Manufacturing Process A General Instructions / Precautions Ensure area and equipment cleanliness before starting the manufacturing operations. Check and ensure that all manufacturing equipments and other required accessories are clean and ready for use. Check calibration of respective equipment / machine before use Wear Gloves and Nose mask during all manufacturing process. Counter check the weights of all ingredients before using in the batch. Get line clearance from QA for manufacturing. AHU system should be kept ON throughout the manufacturing process. Temperature should be kept at 25 0 C ± 2 0 C and Relative Humidity should be kept at 45% ± 5% RH. Check and maintain record of humidity and temperature at least twice in a shift. Ensure that only QC approved Purified Water is being used for manufacturing purpose. Record AR No. During the preparation of this product, no other product processing should be done in the same area. Whatever sifting or filtration through SS mesh is involved, check the mesh integrity before and after use. All critical aspects during manufacturing like temperature, duration of mixing, weight etc. must be checked and recorded by the supervisor. Supervisor to ensure completion of all in-process records during various stages of manufacturing operations till completion of the batch. Release from QA should be taken for all in-process tests mentioned in batch manufacturing record. No over writing is allowed in the Batch Manufacturing Record. If the initial data is wrongly entered, cancel the data by single stroke across and initial. Record reasons for change as foot- note on the same page. All the details whatever is necessary should be recorded in Batch Manufacturing Record and Batch Packaging Record. Send a Test Request Form to QC after manufacturing is completed. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 48 of 124

32 Check all polyethylene bag before and after material loading for black particles. Manufacturing Equipments: Table 2.12 Manufacturing Equipments used during Capsule manufacturing Sr. no. EQUIPMENT NAME 1. Weighing balance 220 g capacity 2. Weighing balance 20 Kg capacity 3. Double Cone Blender 200 Liters 4. Multi mill 5. Double cone Blender 6. Oscillating Granulator 7. Intermediate process containers / drums (IPC) 8. Capsule Filling Machine Ancher 9. Capsule Polishing Machine 10. Capsule Sorter 11. Strip/Blister Packing Machine 12. Packing Conveyor Belt 13. Air Displacement Unit Manufacturing process: a) Weighing and Checking Weights were checked for the raw materials as per the formulation sheet (i.e. page No.3) The AR No. of all the materials was also checked as per the issuance sheet. b) Sieving: Tianeptine Sodium was sieved through #80 and others through # 60 sieve fitted to a Vibrosifter and collected in separate Intermediate Process Containers c) Granulation: Granulation was done with the help of Purified water with the following Raw material in quantities as mentioned herewith. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 49 of 124

33 Table 2.13 Standard quantities of Raw materials used during Granulation Ingredients Spec Standard Quantity (Kg) Tianeptine sodium BP * Microcrystalline cellulose (PH 102) BP Microcrystalline cellulose Plain BP Maize starch BP Magnesium stearate BP Colloidal anhydrous silica BP Purified Talc BP Croscarmellose sodium BP Sodium starch glycolate BP Crospovidone BP d) Blending (Lubrication): All the sieved materials were sifted in Double cone Blender and blended for 30 minutes at 10 RPM. Unloaded the granules in double polythene lined drums and closed tightly. Labeled the drums appropriately and recorded all the details including Product Name, Batch No., Batch size, Mfg. Date. Request slip was sent to QA department to draw the sample for bulk analysis done by QC department. Ensured that temperature and humidity in the Capsule filling area is 25 C± 2 C, NMT 45 %± 5% respectively. Undue storage of granules Avoided. Granules were taken up immediately for filling into Capsule shells. e) In-Process Checking In process parameters at regular intervals were monitored and recorded in the following formats. Q.A. / Q.C. for to do in-process checks as per the norms set by them. Send the sample of blend to QC for Assay. Attached the requisition slip and QC In-process results slip received from QC. After blend was released from QC, bulk was filled into capsules as per specification. Storage Condition :Store below 25 º C, Protect from light Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 50 of 124

34 TEST LIMIT Description Off white granular Powder Assay : Tianeptine sodium BP 98.0 to 102.0% Moisture content NMT 2 % f) Quality Assurance / Quality Control Approval After getting Quality Approval, the Blend was transfered to Capsule Filling dept. and the product was filled as per specifications and instructions g) Filling Ensured that temperature and humidity in the filling area was 25 C± 2 C, NMT 45%± 5% respectively. Undue storage of granules Avoided. Granules were taken up immediately for filling into Capsule shells. The Capsule Filling Machine was set as per respective SOP. In process parameters at regular intervals were monitored and recorded in the following formats. Q.A. / Q.C. for to do in-process checks as per the norms set by them. The filling parameters were adjusted as per the standards given below:- Table 2.7 Capsule Filling parameters as per the standards Parameters Standards Appearance Light green / Light green hard gelatin capsule size 5, containing off white granular powder. Standard Fill Weight of Capsule 93mg + 2 % Weight of Filled Capsule mg + 2 % Weight of 20 Capsules 2.41 g Individual Weight Variation + 10 % Disintegration Time NMT 20 minutes Locking Length of Filled Capsule 10 ± 0.2 mm h) In-Process Checking Frequency In process parameters at regular intervals were monitored and recorded in the following formats. Q.A. / Q.C. for to do in-process checks as per the norms set by them Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 51 of 124

35 Weight of 20 Capsules Fill Variation Disintegration time Locking Length of Filled Capsule Every 30 minutes Every 120 minutes Every 120 minutes Every 120 minutes i) Rejection The Rejection/Recovery of the Blend/Capsules were collected, recorded, labeled appropriately and destroyed as per respective SOP j) Quality Assurance / Quality Control Approval Request slip to Q.A. Department was sent to draw the sample for finished product analysis to be done by QC Dept. After getting Quality Approval, Capsules were transfered the to packing dept. and packed the product as per current packaging specifications and instructions k) Packaging: Batch Issuance Slip from Manufacturing to Packing Table 2.8 Requisition of Packing Material Item code PRIMARY PACKING COMPONENTS Aluminum Blister foil Size 185mm B.P.F7 69 Items UOM Standard quantity Kg B.P.0 18 PVC Film Size 188 mm clear SECONDARY PACKING COMPONENTS B.P. AA2 6 Kg Printed cartons Tianeptine sodium Capsules 10 x 10 Nos 1000 B.P. J01 1 B.P. T76 Corrugated Box G-50 Nos Literature Tianeptine sodium Capsules Nos 1000 B.P. K10 Plastic film 500 mm (20 ) Kg Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 52 of 124

36 l) Batch Details Instructions and Certification Transfer the batch IPCs to the packing cubicle and check if the number of containers received for the batch. Load the IPC in machine hopper in serial order Load the Printed foil on winding shaft of the Blister Packing machine Perform the Leak test on the samples drawn during machine setting as per the relevant SOP. Start the machine only after the Blister passes the leak test and record the observation. Set the sealing heater temperature. Monitor it initially and there after every 60 minutes by Production Department and after every 120 minutes by QA Department. Record the observations in the In Process Sheet. Line Setting Set the packing line After setting all the parameters check and certify the line setup Checker should check individual Strip/Blister Pack 10 Capsules in each blister Pack Such 10 Blisters in a Carton Insert a Literature in each carton Pack such 100 cartons in corrugated box Wrap the Shipper with plastic film 500 mm Seal the shipper using BOPP tape Label the shipper using sticker label. Number each shipper serially on center of the corrugated box. In case any excess packing material is required to complete the batch, get the same on Additional Material Requisition form. Transfer of finished products to quarantine finished goods store: After completion of terminal inspection and getting Quality Assurance approval on the Finished Goods, the entire quantity of Finished Products was transferred to the quarantine finished goods store by Transfer Note. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 53 of 124

37 Control of critical steps and intermediates Table 2.10 Critical Parameters & Acceptance criteria Stage Parameter Acceptance criteria Blending Assay 98.0 to 102.0% Moisture Content NMT 5 % Capsule Filling Average fill 93 mg ± 2% Fill Variation ± 10 % Disintegration time NMT 20 minutes Length 5 ±0.2 mm Chemical Analysis As per Finished Product Specification Microbial Analysis Total bacteria count (NMT 1000 cfu/gm) Total fungal count (NMT100 cfu/gm) Hold Time Study Blend Assay 98.0 to 102.0% Moisture Content NMT 5 % Microbial Analysis Total bacteria count (NMT 1000 cfu/gm) Total fungal count (NMT100 cfu/gm) Capsule Filling Average fill 93 mg ± 2% Fill Variation ± 10 % Disintegration time NMT 20 minutes Length 5 ±0.2 mm Chemical Analysis As per Finished Product Specification Microbial Analysis Total bacteria count (NMT 1000 cfu/gm) Total fungal count (NMT100 cfu/gm) Blister Packing Leak Test None of the strips shall fail in the leak test Acceptance Criteria i) All the tests performed during the process should qualify the specifications. ii) Any deviation from the acceptance criteria of the specific check point shall be reported and decision should be taken for the acceptance, rejection, change in the formulation /process Process Validation and/or Evaluation a) Objective: To validate the manufacturing process of Tianeptine Sodium Capsule 12.5 mg as per the Batch Manufacturing Record to establish documentary evidence that the process will consistently producing the product meeting its in-process & finished product specifications by examining three consecutive production batches Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 54 of 124

38 b) Scope: Retrospective validation was carried out for 3 consecutive production batches for Tianeptine Sodium Capsule 12.5 mg ; critical steps were monitored and validated. The objective of this process validation report for Tianeptine Sodium Capsule 12.5 mg was to establish documented evidence that the process was capable of consistently producing a product of predetermined specifications and quality attributes. c) Batches Used for Validation Table 2.14 Batches used for validation Batch No Batch Size Mfg Date Exp Date BC Capsules 05/12/ /11/2012 Batch No Batch Size Mfg Date Exp Date BC Capsules 17/12/ /11/2012 Batch No Batch Size Mfg Date Exp Date BT Capsules 22/12/ /11/2012 d) In-Process Checking The sample of blend was sent to QC for Assay. The requisition slip was attached and QC In-process results slip received from QC. After release of blend from QC, the bulk was filled into capsules as per specification. Storage Condition :Store below 25 º C, Protect from light TEST LIMIT Description Off white granular Powder Assay : Tianeptine sodium BP 98.0 to 102.0% Moisture content NMT 2 % e) In-Process Checking Frequency In process parameters at regular intervals were monitored and recorded in the following formats. Q.A. / Q.C. for In-process checks as per the norms set by them. Weight of 20 Capsules Every 30 minutes Fill Variation Every 120 minutes Disintegration time Every 120 minutes Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 55 of 124

39 Locking Length of Filled Capsule Every 120 minutes f) Sampling plan: Sampling Locations for Double Cone Blender After completion of mixing for the defined period of time, the samples were collected from different locations. Top left (T7-1) Top right (T7-2) Middle left (M7-1) Middle right (M7-2) Bottom left (B7-1) Bottom right (B7-2) 20 g (10 g from left + 10 g from right) sample was collected from each sampling point perform the same activity with the time interval of 5, 10, 15 minutes. (T7, T8, T9). g) Validation Sampling Plan Table 2.15 Sampling Plan Validation Unit Sampling point Operation Blending Sample was drawn from 9 locations of the blender interval of 15,25,30 min IPC Sample 40 g composite sample from Top, middle, bottom layers after unloading the material into intermediate process containers. Blend Hold 20 g (2 20) g (for Time Study chemical and microbial limit tests) composite sample Sample quantity 20 g each from each sampling location Sample code L-1 M-2 R-1 L-2 M-2 R- 2 L-3 M-3 R-3 Controlled parameter RPM of the blender Blending time Measured response Assay Identify Moisture content 40 g S1 --- Assay Moisture content 40 gm (20 gms for chemical analysis and,20 gm for microbial limit tests) S2-A, S2-B --- Assay Moisture Content Microbial tests Acceptan ce criteria Assay to 102.0% Water NMT 5 % Assay to 102.0% Water- NMT 5% Assay to 102.0% Water- NMT 5% Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 56 of 124

40 Capsule Filling Filled capsules Hold Time Study Blister packi ng Capsules after every 2 hours, 20 Capsules after every 30 minutes, 80 capsules composite sample for Finished Product Analysis Caps(50 Caps) for chemical analysis + 30 Caps for microbial limit tests) Strips were withdrawn after every 2 hrs intervals for leak testing (no of strips that are taken for leak testing shall represent each cavity of the sealing rollers). --- S Vacuum, Air, Speed, Height of Powder in Hopper Average fill Fill Variation DT S Chemical Analysis Microbial Analysis S Assay Moisture Content DT Microbia l Tests Sealing/ roller temperature Avg fill 93mg±1 0% Fill Variation 10% DT: NMT 20 min Assay to 102.0% Water- NMT 5% Assay to 102.0% Water- NMT 5% DT: NMT 20 min Leak test None of the strips were failed in the leak test h) Tabulation of the Test Results for following batches: Table 2.14 Batches used for validation Batch No Batch Size Mfg Date Exp Date BC Capsules 05/12/ /11/2012 Batch No Batch Size Mfg Date Exp Date BC Capsules 17/12/ /11/2012 Batch No Batch Size Mfg Date Exp Date BT Capsules 22/12/ /11/2012 i) Evaluation of Data, and Where Applicable, including Statistical Process Control Analysis: Refer to Manufacture in Chapter 3.RESULT & DISCUSSION. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 57 of 124

41 j) Batch Analyses Table 2.16 Batch Analyses for BRUTINE (Tianeptine sodium Capsules 12.5 mg) Test BC BC BC Limits Light green- Light green Hard gelatin Light green- Light green Hard gelatin Light green- Light green Hard gelatin Light green- Light green Hard gelatin Capsules, Description Capsules, Size Capsules, Size Capsules, Size Size 5.Containing 5.Containing 5.Containing 5.Containing off white granular powder. off white granular powder. off white granular powder. off white granular powder. Positive Positive Positive Positive for Identification Tianeptine sodium Average fill Weight 93.6 mg 93.8 mg 93.4 mg 93.5 mg ±5.0 % Uniformity Weight Dissolution Assay: Each Capsule contains- Tianeptine sodium BP 12.5 mg of mg± % minutes mg ± % 28-35minutes mg ± % minutes 99.93% 99.21% 99.44% ± % of the average weight NLT % of the labelled amount of Tianeptine sodium dissolved in 45 min. Claim 12.5 mg % k) Results Statement was made for in this report on acceptability of manufacturing process meet the objective of protocol Control of Excipients Specifications The excipients Microcrystalline Cellulose BP, Purified Talc BP, Magnesium Stearate BP, Maize Starch BP, Sodium Starch Glycolate BP, Colloidal Anhydrous Silica BP, Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 58 of 124

42 Croscarmellose Sodium BP, Crospovidone BP and Purified Water BP were procured as official specifications. Protocol (For all compendial Excipients): British Pharmacopoeia 2011 Hard Gelatin was obtained as In-house specifications. PROTOCOL: In-house HARD GELATIN CAPSULES Table 2.17 Specification for Hard Gelatin Capsule Shells Sr. No. Tests Specification 1 Description Light green/ Light green, Empty Hard Gelatin, Size 5 Capsules. 2. Odour Shell should not develop any foreign odour. 3 Identification A precipitate should produce. 4 Average Weight 27.5 mg ± 10% w/w. 5 Dimension Outside Diameter of Cap Outside Diameter of Body Length of Cap Length of Body Joined Length mm mm mm mm mm 6 Disintegration time Not more than 15 minutes. 7 Loss on drying Between 12.5% to 16.0 % determined on 1.0 gm sample 8 Microbial Contamination Total microbial count: Not more than 1000 CFU per gm. 1.0 gm is free from Escherichia coli and Salmonellae. spp Analytical Procedures Hard Gelatin Capsules shell Description: Light green/ Light green, Empty Hard Gelatin, Size 5 Capsules. Odour: Shell was not develop any foreign odour. Identification: One capsule shell was boiled with 20 ml of water, allowed to cool and centrifuged. To 5 ml of the supernatant liquid, 1 ml of picric acid solution was added and to another 5 ml, 1 ml of tannic acid solution was added; a precipitate was produced in each case. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 59 of 124

43 Average weight: 100 capsule shells were weighed and the average weight was determined of a capsule. The average weight was within +/-10% of the target weight. Disintegration time: The capsule shells were complying with the disintegration test for tablets and capsules, using the discs. The capsules were disintegrated within 15 minutes. Loss on drying: Loss on drying was determined by drying of 1.0 g of capsule shells in an oven at 105 o for 4 hours / constant weight. Value lies between 12.5 and 16.0%, Microbial Contamination: Total microbial count: Not more than 1000 CFU per gm. 1.0 gm is free from Escherichia coli and Salmonellae. spp Validation of Analytical Procedures As all the Excipients were present in the official monograph i.e. B.P. and EP, So there was no requirement for the analytical method validation Justification of Specifications As all the Excipients were present in the official monograph i.e. B.P. and EP, so there was no requirement for the Justification of Specifications Excipients of Human or Animal Origin Gelatin was used in Capsule shell manufacturing was BSE free and was conferred by a BSE risk free certificate from the Capsule shell manufacturer. Find attachment in Annexure 1 as Certificates Novel Excipients Not Applicable Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 60 of 124

44 Control of Drug Product Specification Table 2.18 Specification for BRUTINE (Tianeptine sodium Capsules 12.5 mg) Product Name: BRUTINE (Tianeptine sodium Capsules 12.5 mg) Mfg Date: 12/2012 Batch No: BC Exp Date: 11/2014 Quantity: 40 Capsules Date of Analysis: 05/12/2012 Test Specification Result Description Light green-light green Hard Light green-light green gelatin Capsules, Size Hard gelatin Capsules, 5.Containing off white granular Size 5.Containing off powder. white granular powder. Identification Positive with Test A and B Positive Average fill Weight 93.5 mg ±5.0 % 93.6 mg Uniformity of Weight ± % of the average weight mg± % Ethyl acetate(ppm) NMT Individual impurity: Not more than 0.2 % 0.081% Total Impurities NMT 1.0 % 0.24% Dissolution Assay: Each Capsule contains: Tianeptine sodium BP 12.5 mg NLT % of the labelled amount of Tianeptine sodium dissolved in 45 min. Claim 12.5 mg % minutes 99.93% Specification of Tianeptine sodium Capsules 12.5 mg was In-house & the test & methods were validated as per the analytical method validation, stability study & other testing as dissolution. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 61 of 124

45 Analytical Procedures Method of Analysis Details of Tianeptine on HPLC 5. Column:- C Mobile Phase: - Mobile phase consist of acetonitrile: 50 mm potassium dihydrogen phosphate, ph 3.5 (33:67, v/v/). 7. Flow rate: - It having pump at a flow rate of -1ml/min. 8. λ max: - Operating at -214 nm. Standard stock solutions Standard stock solutions of 100 µg/ml was prepared in mobile phase by dissolving the pure drug in it. Preparation of the calibration curves Aliquots were prepared by serial dilution method sufficient to produce ng/ml. Each solution was injected one by one and chromatogram was recorded at 214 nm. The peak area of drug was noted and calibration curve was plotted as peak area against concentration of drug. Analysis of capsule formulation The contents of 20 capsules were mixed and weight accurately. Powder equivalent to specification transferred into a 100 ml volumetric flask, dissolved in water and sonicated for 5 min., the volume was made up with water, shaken well for 5 min. and then filtered, further absorbance value was noted in triplicate at 214 nm against reagent blank. Instrumental Conditions: Instrument: UV-VIS Spectrophotometer (Shimadzu make Model-1800) Wavelength: 214 nm Procedure The absorbance of the test solution was measured by using the UV-VIS Spectrophotometer at the maximum at 214 nm and record the spectra was recorded. Calculation: The content was calculated taking 562 as the value of A (1%, 1 cm) at the maximum at 214 nm. Test Abs potency = x x x x x Test wt Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 62 of 124

46 Validation of Analytical Procedures By: UV-visible double beam spectrophotometer Refer to Annexure 2 Chromatograms & Testing Procedure By: HPLC: (a) Objective: This report is intended to perform the experiments and established the validity of the titled method for its intended use for the determination of assay content of Tianeptine sodium capsules 12.5 mg. The assay method was developed and verified for its intended purpose. (b) Scope: This method validation report is applicable to the determination of the assay of Tianeptine sodium capsules 12.5 mg. (c) Responsibility a. Quality Control : i. To prepare the method validation report ii. To evaluate the results against the acceptance criteria iii. To review the report for its compliance b. Quality Assurance i. To approve the method validation report. (d) Instrument used i. UV-VIS Spectrophotometer ii. Ultra sonic bath iii. Analytical Balance iv. HPLC (e) Reagent Used i. Ethyl Acetate ii. 0.5 M Sulphuric Acid iii. Tianeptine WS Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 63 of 124

47 (f) Method of Analysis Details of Tianeptine on HPLC 9. Column:- C Mobile Phase: - The Mobile phase consist of acetonitrile: 50 mm Potassium dihydrogen phosphate, ph 3.5 (33:67, v/v/). 11. Flow rate: - It having pump at a flow rate of 1ml/minute. 12. λ max: - Operating at -214 nm. Standard stock solutions A standard stock solution of 100 µg/ml was prepared in mobile phase by dissolving the pure drug in it. Preparation of the calibration curves Aliquots were prepared by serial dilution method sufficient to produce ng/ml. Each solution was injected one by one and chromatogram was recorded at 214 nm. The peak area of drug was noted and calibration curve was plotted as peak area against concentration of drug. Analysis of capsule formulation The contents of 20 capsules were mixed and accurately weight the content of 20 Capsules. Powder equivalent to specification transferred into a 100 ml volumetric flask, dissolved in water and sonicated for 5 min., the volume was made up with water, shaken well for 5 min. and then filtered, further absorbance value was noted in triplicate at 214 nm against reagent blank. Instrumental Conditions: Instrument: UV-VIS Spectrophotometer (Shimadzu make Model-1800) Wavelength: 214 nm Procedure The absorbance of the test solution was measured by using the UV-VIS Spectrophotometer at the maximum at 214 nm and the spectra was recorded. Calculation: The content was calculated taking 562 as the value of A (1%, 1 cm) at the maximum at 214 nm. Test Abs potency = x x x x x Test wt Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 64 of 124

48 (g) Specificity Acceptance Criteria: No any interference observes at maximum 214nm from the diluents and placebo maxima. Procedure: Blank solution, placebo and test solution by using Tianeptine sodium capsules were prepared as per method of analysis and taken the absorbance of the test solution and calculated the measured the peak response of the major maxima. (h) System Suitability Acceptance Criteria: a) The tailing factor for Tianeptine Sodium peak is not more than 2.0. b) The column efficiency determined from the Tianeptine Sodium peak is not less than 2500 theoretical plates. c) The % RSD for three replicates absorbance of test solution is not more than 2%. Procedure: Blank solution and test solution (concentration: 1 mg/ml) were prepared as per method of analysis and the maximum was observed at 214 nm and calculated the % RSD for the three replicates absorbance of the test solution. (i) Method precision Acceptance Criteria: % RSD of the assay content from the six sample preparation should be not more than 2.0% Procedure The method passed the test for repeatability as determined by %RSD of the replicates absorbance of the test solution at 100% test concentration. (j) Accuracy Acceptance Criteria: % Recovery should be between 95% and 105 % and % RSD of the 9 determination of the samples should be not more than 2.0% Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 65 of 124

49 Tianeptine API was spiked in our Capsule. The result of accuracy given in tabulated below revealed that the method was found accurate for all above purposes. Table 2.19 % Recovery of Tianeptine at different sample concentration Sr. No. Sample Concentration (in %) of the Target conc. Sample ID 1 Sample Preparation -1 2 Sample 75% Preparation -2 3 Sample Preparation -3 4 Sample Preparation -4 5 Sample 100% Preparation -5 6 Sample Preparation -6 7 Sample Preparation -7 8 Sample 125% Preparation -8 9 Sample Preparation -9 % Recovery of Tianeptine % Recovery Mean : 99.97% RSD : 0.789% (k) Limit of detection & Limit of quantitation: LOD (Limit of detection): LOD is X + (3 x SD). LOD (Limit of detection) value - 10 ng/ml LOQ (Limit of quantitation) LOQ is X + (10 x SD). LOQ (Limit of quantitation) value - 45 ng/ml Ruggedness: Ruggedness of an analytical method was checked by performance the method used for precision on different days by different analyst with different regent. The ruggedness data was found within the acceptance limit NMT 2.0% RSD. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 66 of 124

50 (l) Conclusion The determination of assay content of Tianeptine in Tianeptine sodium capsules; was verified by using the HPLC. On basis of the analytical data and results it was found that method was specific, precise, Accurate and system suitability parameters observed well within the pre-defined acceptance criteria. Hence it was concluded that method for the determination of assay of Tianeptine in Tianeptine sodium capsules was suitable for our intended purpose Batch Analyses Refer to Batch Analyses in Chapter 3. RESULT & DISCUSSION Characterisation of Impurities IMPURITIES A. Br - [CH 2 ] 6 - CO- O -C 2 H 5 : ethyl 7 - bromoheptanoate, B. R = H, R 1 = [CH 2 ] 6 - CO- O- C 2 H 5 : ethyl 7- [[(11RS) -3- chloro- 6-methyl -6,11- dihydrodibenzo [c,f] [1,2] thiazepin-11-yl]amino] heptanoate S, S-dioxide, E. R = R 1 = [CH 2 ] 6 -C O 2 H: 7, [[(11RS) -3- chloro- 6- methyl-6,11- dihydrodibenzo [c,f] [1,2] thiazepin 11 - yl] imino] diheptanoic acid S, S-dioxide, C. X = O: 3 -chloro -6-methyldibenzo [c,f] [1,2] thiazepin- 11 (6H)- one S,S-dioxide, D. X = N - [CH 2 ] 6 -CO 2 H: 7- [[ (11RS)-3- chloro-6-methyldibenzo [c,f] [1,2] thiazepin-11 (6H) -ylidene] amino] heptanoic acid S,S-dioxide. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 67 of 124

51 Justification of Specification(s) Table 2.20 Justification of Specification BRUTINE (Tianeptine sodium Capsules 12.5 mg) Description Test Limits Result BC Light green-light green Hard gelatin Capsules, Size 5.Containing off white granular powder. Light green-light green Hard gelatin Capsules, Size 5.Containing off white granular powder. Positive Specification IHS Identification Positive with Test A IHS and B Average fill 93.5 mg ±5.0 % 93.6 mg IHS Weight Uniformity of ± % of the IHS mg± % Weight average weight Ethyl IHS NMT acetate(ppm) Individual 0.080% IHS Not more than 0.2% impurity: Total Impurities NMT 1.0% 0.24% IHS NLT % of the IHS Dissolution labelled amount of Tianeptine sodium dissolved in 45 min minutes ASSAY: 99.93% IHS Each Capsule Claim 12.5 mg contains % Tianeptine sodium BP 12.5 mg The testing Procedure was justified from data of analytical method validation and the other methods like dissolution and other In-process results were justified from process validation and Stability study upto the shelf life of Product. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 68 of 124

52 Reference Standards or Materials Given below was the detail of working standard used for analyzing the samples of Capsules and API. The IR spectra of the working standard was also attached. Table 2.21 Reference standard/materials Name Batch no. Assay Tianeptine Sodium BP BP/BPA_001/D/10/ % w/w TEST STANDARD RESULT Description White to yellowish powder, very Off-white powder, hygroscopic. hygroscopic Identification Should Be Complies Complies Water NMT 5.0%W/W 0.55% Assay NLT 99.00% and NMT % W/W (O.A.B) 99.65% w/w Refer to Annexure 2 for Certificate of Analysis Container Closure System Primary Packing Components Printed Aluminium foil PVC Secondary Packing Components Carton Tertiary Packing Components The cartons were packed in a corrugated fiberboard box (depending on the type of packing details). All the cartons were poly-laminated in a group of 10 cartons,by which the cartons were protected from humidity and other damages. The corrugated fiberboard boxes were made of export worthy material and were sealed with a BOPP tape. Transportation: Transported with precautions. The cautions like - Not for loose handling Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 69 of 124

53 Protect from water Avoid vigorous transportation Care should be taken. SPECIFICATION OF PACKING MATERIAL PRESENTATION FORM: 10 x 10 blister pack in a unit carton along with pack inserts; such cartons were packed in a corrugated box sealed with cello tape and labeled. PRIMARY PACKAGING MATERIAL: Table 2.22 Specifications for Printed Aluminium foil Sr. No. Tests Limits / Requirements 1. Description i) Print color Printed aluminium blister foil with VMCH coating. As per artwork / standard. 2. Width mm 3. Thickness mm 4. Total GSM 69.34GM/CM 2 5. Aluminium GSM 66.53GM/CM VMCH GSM Ink Lifting Test Suitability Defects Supply Storage 2.81GM/CM 2 Should passes the test 1. Winding of rolls not to be loose / telescopic. 2. Rolls should be free from wrinkles, creasing, / dents and collapsed / damaged core 3. Ink used for printing should be HR grade and withstand temp. up to 180 C, there should not be lifting or fading of ink during blister sealing Sample size of 5% rolls to be inspected at random for corroded or oxidized surface, improper coating, and improper winding and incorrect diameter of roll. Rolls wrapped in polythene film with brawn paper. Rolls should properly label with quantity, item name, and supplied by. Storage: Protect from light and moisture at a temperature < 30 C Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 70 of 124

54 Table 2.23 Specifications for Clear PVC Film Sr. No. Tests Specification 1 Description A clear, smooth Poly Vinyl Chloride film 2 Width 188 ± 1 mm 3 Grammage 340 gm/m 2 ±8 % 4 PVC Thickness mm ± 0.02 % 5 Pin Holes Should be absent 6 Colour As per specification 7 Inner Dia Core 75 ± 1mm Table 2.24 Specifications for Cartons (10x10 s) Sr. No. Tests Specification 1. Description i) Color ii) Shape iii) Material of Construction As per artwork / standard. Rectangular, side opening, reverse tuck Printed / unprinted, duplex board. 2. Dimension: (Outer) i) Length ii) Breadth iii) Width As per approved sample As per approved sample As per approved sample 3. Grammage 300g ± 10% of GSM (For duplex board carton) Printed Matter Suitability Supply Storage Sharp and elegant as per artwork The inks used for printing should be scuffproof and rub resistant. Cartons should pass the test when checked on scuff-proof tester. Banded in bundles of 100 nos. and such 10 bundles wrapped in brown paper clearly marked with item name, quantity and supplied by. Storage: Protect from light and moisture at a temperature < 30 C. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 71 of 124

55 Table 2.25 Specifications for BOPP Tape (Printed Self Adhesive Tape) Sr. No. Tests Specification 1. Description Transparent self-adhesive BOPP tape printed as per approved text and design in blue color. 2. Dimension: i) Width ii) Thickness of BOPP iii) Total Thickness 63 ± 1 mm microns microns 3. Grammage 50 g ± 10% Core Diameter Internal Adhesive Adhesion test Supply Storage 75 ± 1 mm Should be uniform, printed surface should pass tape test. Seal the flaps in length on corrugated fiber box the tape using a hand dispenser or manually. Ensure no wrinkles/air bubbles are formed while pasting and proper pressure should be applied. Leave the box for 10 minutes and try to separate the tape from the end. Tape should separate with fiber tearing. Supplied in rolls of 65 meters packed in a box and clearly marked with item name, quantity and supplied by. Storage: Protect from light and moisture at a temperature < 30 C. Table 2.26 Specifications for Corrugated Box G -56 Sr. No. Tests Specification 1 Description 2 Width Length Width Height A Brown Craft Paper Corrugated box G-56 with 5 ply. 480 mm ± 5mm 348 mm ± 5mm 347 mm ± 5mm 3 Grammage NLT 630 gm/m 2 4 Bursting Strength N.L.T 8 kg/cm 2 5 No of ply 5 6 Type of Fluting B 7 Joint Stapled/Pasted Should comply 8. Moisture content 7% to 10%. 9 Text Matter As per approved art work. 10 Printing and colour scheme As per approved art work. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 72 of 124

56 Stability Stability Summary and Conclusion Refer to Stability in Chapter 4 SUMMARY AND CONCLUSION. Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 73 of 124

57 Shri Jagdish Prasad Jhabarmal Tibrewala University, Jhunjhunu, (Raj) Page 74 of 124