Expression System and Process Development for MIS Scale Up

描述(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例新病例和大约13,850例死亡。在大多数患者中，癌症在最初治疗后复发，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，来自马萨诸塞州总医院(MGH)的Donahoe、MacLaughlin和Teixeira博士表明，重组人M？llerian抑制物质(RhMIS)针对对当前使用的化疗药物反应差的卵巢癌细胞群体，可能成为一种创新的靶向药物，以治疗复发的多药耐药卵巢癌。在体内，当作为单一药物测试时，发现它在持续80至100天的实验中抑制了肿瘤的生长。此外，由于MIS受体在内源性组织中的表达很低，预计重组人MIS的全身毒性较低。阻碍重组人巨噬细胞集落刺激因子治疗卵巢癌进展的一个重要的未得到满足的需求是以合理经济的方式发展规模化生产蛋白质。该R43方案描述了在荧光假单胞菌中成功表达重组人巨噬细胞集落刺激因子的平行分子生物学处理方法，以及在荧光假单胞菌中规模化生产重组人巨噬细胞集落刺激因子所需的研究过程。来自Nemucore医疗创新公司(NMI)、Pfenex和MGH的团队建议作为R43申请的主题，全面开发和表征使用一次性生物制造设备生产rhMI所需的方法和流程。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，以支持生产用于临床研究的新型生物疗法，如重组人免疫缺陷病毒。

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.