P128 An Approach to Control Purity of the Drug Substance for Non-Clinical Study T. HASHIMOTO1, Y. KOBAYASHI1, Y. OKUBO*1 1 Res. Lab., Toyama Chemical Co., Ltd., Toyama, JAPAN Synthetic Route of API (Compound 12) *Corresponding author: Yusuke Okubo, Mailing address: Research Laboratories, TOYAMA CHEMICAL CO., LTD. 2-4-1 Shimookui Toyama 930-8508, JAPAN, E-mail: YUSUKE_OKUBO@toyama-chemical.co.jp Tel: +81 (0)76-431-8267 Fax: +81 (0)76-431-8208 Background: Purity of drug substance (API) used for clinical study must be the same or higher than that of API used for non-clinical study (NCAPI). It is important to allow flexibility in developing manufacturing process to design and control the purity of NCAPI. There are various ways to optimize the purity on purpose, such as addition of individual impurities on use of harsh chemical reactions. In the present study, we established a method to ensure the desired purity of NCAPI by adding a separate set of API containing various species of impurities (IMAPI). Methods: Firstly, the target purity of NCAPI was determined by the purity of API used for a two-week repeated dose toxicity study in the dog. Secondly, IMAPI was prepared in lab, and lab experiments were conducted to determine how much IMAPI needed to achieve the target purity of NCAPI at large-scale manufacturing. Thirdly, we conducted large- scale manufacturing by addition of IMAPI and confirmed whether the purity of NCAPI was within the desired one. Results: The purity of API used in the dog toxicity study was 97.8%, so we set the target purity at 98%. IMAPI was synthesized with the purity of 82.2%, and added to the reaction mixture in large-scale production. Crystallization and filtration gave 37.4 kg of NCAPI with the purity of 99%, which is slightly higher than the target purity. Conclusions: These results indicate that this method is useful for purity design of NCAPI. Abstract Introduction Methods 1. Target purity and impurity profile determination API for a two-week repeated dose toxicity study in the dog was synthesized. Target purity and impurity profile of NCAPI were determined. 2. Preparation of IMAPI IMAPI was prepared by adding the residue of the filtrates, process intermediates, and degradate, which were prepared separately. 3. IMAPI spike tests Reaction mixture in the final manufacturing step was spiked IMAPI to determine where the appropriate process to add and how much IMAPI needed to optimize the purity and the target impurity profile of NCAPI at large-scale manufacturing. 4. Manufacturing NCAPI NCAPI was manufactured by adding IMAPI to the reaction process of the final manufacturing step. Result 2: Preparation of IMAPI Result 1: Synthesis of the API for a two-week repeated dose toxicity study in the dog Summary of results 1. The purity of API used in the dog toxicity study was 97.8%. Therefore, the target purity of NCAPI was set at 98%. Major impurities in this API were identified to determine the target impurity profile. 2. IMAPI was synthesized with the purity of 82.2% and major impurities were included. 3. Spike tests showed that adding 1.0 wt% IMAPI to the reaction process produced API with the purity of 98.0%, achieving the target purity profile. Adding IMAPI to the crystallization process did not affect the purity of the API. 4. NCAPI was manufactured by IMAPI. Adding IMAPI to the reaction process afforded 37.4 kg of NCAPI with the purity of 99.0%, which is slightly higher than the target purity. Impurities presented in NCAPI were as we anticipated. Conclusion Alkylation Reaction 75% yield Pd-Coupling Reaction 90% yield Ring-Closing Reaction Reaction with Hydroxylamine 80% yield 95% yield Coupling Reaction 90% yield APIs intermediates filtrates byproducts degradates + = IMAPI Result 3: Spike tests of IMAPI Result 4: Manufacturing NCAPI Ring-Closing Reaction Work-up Crystallization Filtration Compd. 11 Crude Compd. 12 Filtrate Dissolution Crystallization Filtration and Wash Filtrate Compd. 12 450 g (two lots)