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21，860例新发病例和约13，850例死亡。绝大多数患者在初始治疗后癌症复发，并表现出对传统和新型化疗药物的耐药性，这代表了一个巨大的临床挑战，可以通过生产新型靶向治疗来缓解。最近，来自马萨诸塞州总医院（MGH）的Donahoe，MacLaughlin和特谢拉博士表明，重组人M？勒氏抑制物质（rhMIS）特异性靶向卵巢癌细胞群，这些细胞群对目前使用的化疗药物反应较差，可能是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作为单一药剂进行测试时，发现其在持续80至100天的实验中抑制肿瘤生长。此外，由于MIS受体在内源性组织中表达不足，因此预计rhMIS的全身毒性较低。阻碍卵巢癌rhMIS治疗进展的一个显著未满足的需求是以合理的经济性开发蛋白质的规模化生产。本R43提案描述了创建>1000株菌株和质粒组合以在荧光假单胞菌中成功表达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的团队建议将此R43申请作为主题，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制造设施，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.