Expression System and Process Development for MIS Scale Up

描述（由申请人提供）：卵巢癌是一种高度致命的妇科癌症，在美国每年估计有21860例新病例和大约13850例死亡。绝大多数患者，癌症在初始治疗后复发，对传统和新型化疗药物表现出耐药性，这是一个巨大的临床挑战，可以通过生产新的靶向治疗来缓解。最近,Drs。来自马萨诸塞州总医院（MGH）的Donahoe， MacLaughlin和Teixeira证明重组人M？雷氏杆菌抑制物质（rhMIS）专门针对对目前使用的化疗药物反应不佳的卵巢癌细胞群，可能成为一种创新的靶向药物，用于治疗复发性多药耐药卵巢癌。在体内，当MIS作为单一药物进行测试时，在持续80至100天的实验中发现它可以抑制肿瘤生长。此外，由于MIS受体在内源性组织中表达较差，因此rhMIS的系统毒性预计较低。阻碍卵巢癌rhMIS治疗进展的一个重要未满足的需求是合理经济地大规模生产蛋白质的发展。本R43提案描述了在荧光假单胞菌中创建bbb1000菌株和质粒组合以成功表达rhMIS的平行分子生物学处理方法，以及在荧光假单胞菌中大规模生产rhMIS所需的研究工艺开发程序。来自Nemucore Medical Innovations （NMI）、Pfenex和MGH的团队提议作为R43申请的主题，以充分开发和表征使用一次性生物制造设备生产rhMIS所需的方法和工艺。NMI正在建立一个符合FDA药品质量体系指南的模块化生物生产设施，以生产用于临床研究的新型生物治疗药物，如rhMIS。

DESCRIPTION (provided by applicant): Ovarian cancer is a highly lethal gynecologic cancer with an estimated 21,860 new cases and approximately 13,850 deaths in the USA per year. A large majority of the patients, cancer reoccurs after initial treatment and manifest resistance to conventional and novel chemotherapeutic agents representing a formidable clinical challenge, which could be mitigated by the production of novel, targeted therapies. Recently, Drs. Donahoe, MacLaughlin and Teixeira from Massachusetts General Hospital (MGH) showed that recombinant human M?llerian Inhibiting Substance (rhMIS) specifically targets ovarian cancer cell populations that respond poorly to the currently used chemotherapeutic agents and could be an innovative, targeted medicine to address recurrent multidrug resistant ovarian cancer. In vivo when MIS was tested as a single agent it was found to inhibit tumor growth in experiments lasting 80 to 100 days. Additionally since the MIS receptor is poorly expressed in endogenous tissue systemic toxicity of rhMIS is expected to be low. A significant unmet need hindering the progress of an rhMIS treatment for ovarian cancer is the development of scaled production of the protein with reasonable economics. This R43 proposal describes the parallel molecular biology processing methods to create >1000 strain and plasmid combination to successfully express rhMIS in Pseudomonas fluorescens and the investigative process development procedures required to scale production of rhMIS in P. fluorescens. The team from Nemucore Medical Innovations (NMI), Pfenex, and MGH propose as the subject of this R43 application to fully develop and characterize the methods and processes required to produce rhMIS using single use disposable biomanufacturing equipment. NMI is establishing a modular biomanufacturing facility compliant with the FDA's pharmaceutical quality system guidance to enable the production of novel biotherapeutics for clinical investigation, such as rhMIS.