Development of Superactive Analogs of FSH for Human Infertility

开发治疗人类不孕症的 FSH 超级活性类似物

• 批准号: 8511366
• 负责人: Bruce Dale Weintraub
• 金额: $ 97.21万
• 依托单位: TROPHOGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-13 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8511366
• 关键词: Adverse effects; Affect; Affinity; American; Assisted Reproductive Technology; Beds; Binding; Biological Assay; Bioreactors; Carbohydrates; Cattle; Cell Line; Chinese Hamster Ovary Cell; Clinical Trials; Couples; Development; Doctor of Philosophy; Dose; Drug Kinetics; Embryo; Embryo Transfer; Engineering; Family suidae; Follicle Stimulating Hormone; Follicle Stimulating Hormone Receptor; Funding; Generations; Genetic; Glycoproteins; Grant; Guidelines; High Pressure Liquid Chromatography; Hormones; Human; Human Cell Line; Human Development; Human Follicle Stimulating Hormone; Immunoassay; In Vitro; Infertility; Injection of therapeutic agent; Instruction; Investigational New Drug Application; Licensing; Macaca mulatta; Marketing; Metabolic; Methods; Modeling; Monkeys; National Institute of Child Health and Human Development; Newborn Infant; Oocytes; Oryctolagus cuniculus; Ovarian; Performance; Pharmaceutical Preparations; Pharmacodynamics; Phase; Preparation; Production; Program Development; Property; Rattus; Recombinant Follicle Stimulating Hormone; Recombinants; Regimen; Research; Research Personnel; Rodent; Rodent Model; Small Business Innovation Research Grant; Specificity; Superovulation; Technology; Technology Transfer; Therapeutic; TimeLine; Toxicology; United States National Institutes of Health; Weight; Woman; Work; absorption; analog; attenuation; base; blastocyst; cell bank; commercialization; comparative efficacy; good laboratory practice; hormone analog; immunogenicity; improved; in vivo; manufacturing facility; meetings; nonhuman primate; novel; older patient; pre-clinical; preimplantation; programs; pup; receptor binding; receptor expression; reproductive; response; scale up; screening; urinary

DESCRIPTION (provided by applicant): Infertility affects about 10% of American couples, and there is a very large and rapidly growing market for therapeutics in this field, particularly the primary hormone responsible for ovarian oocyte development, human (h) follicle-stimulating hormone (FSH). Although available urinary and recombinant hFSH products have been quite successful, there is currently an unmet therapeutic need for improved FSH analogs for all infertile women and particularly for older patients and those relatively unresponsive to current therapies. We have previously described the first superactive analogs of glycoprotein hormones that considerably increase receptor binding affinity as well as both in vitro and in vivo biopotency and maximal efficacy. During our successfully completed phase 2 SBIR study we have achieved or exceeded all the aims including screening additional FSH analog candidates and identifying the optimal FSH analog TR 4401 for clinical trials, using multiple in vitro and in vivo rodent bioassay models including the classic ovarian weight response as well as those of oocytes, blastocysts and newborn pups resulting from embryo transfer. In all these rodent models 4401 greatly outperformed all currently available recombinant FSH preparations both in potency and efficacy related to both quantitative endpoints as well as qualitative endpoints related to oocyte or embryo quality. In addition we have shown such superior efficacy of TR4401 to standard FSH in human cell lines with reduced FSH receptor expression representing two models of human infertility, in two bovine models of infertility and in Rhesus monkeys using methods emulating human assisted reproductive technology. We have also shown that one injection of TR4401 in cows could produce comparable superovulation to the standard 8-injection regimen of porcine FSH with no attenuation of response after repeated administration. These nonhuman primate and bovine studies showed no evidence of ovarian hyperstimulation by TR4401 at neither a presumably maximal dose, nor any evidence of immunogenicity, the only two side effects of concern to FDA for this minimally modified, and thus presumably safe, FSH analog. Using an HPLC-validated immunoassay we have discovered superior pharmacokinetic properties of TR4401 in comparison to standard FSH both in rodents and monkeys, apparently the result of delayed absorption. We have also achieved development of a stable Chinese Hamster Ovary (CHO) cell line producing high levels of the final TR4401 analog; optimization of large scale bioreactor production methods; development of novel, high capacity purification methods suitable for commercial scale-up; rigorous quantification and characterization of purified analogs by multiple physicochemical methods including carbohydrate analysis. In the current application, following specific directives from FDA obtained in our highly successful Pre-IND Meeting with them, we propose all steps, including specific timelines indicated on a detailed Gantt chart, required by FDA for further commercial development of this novel FSH analog. These include establishment and characterization of a master and working CHO cell bank; manufacturing of two additional large batches of TR4401 (>200 mg each): the first under GLP and the second under GMP compliant conditions; performance all FDA-required efficacy, specificity, stability, metabolic, pharmacokinetic, pharmacodynamic and analytic studies; performance of all FDA-required toxicology studies including two-generation reproductive toxicology assessment in rats and rabbits; and submission of IND to initiate clinical trials. We have licensed TR4401 to the worldwide leading veterinary superovulation company, Bioniche, Inc. for veterinary use, with a possible option to also co-develop the analog for human use.

不孕症影响约10%的美国夫妇，并且该领域的治疗剂市场非常大且快速增长，特别是负责卵巢卵母细胞发育的主要激素，人（h）卵泡刺激素（FSH）。虽然可用的尿和重组hFSH产品已经相当成功，但目前对于所有不孕妇女，特别是老年患者和对当前疗法相对无反应的患者，对改良FSH类似物的治疗需求尚未得到满足。我们先前已经描述了糖蛋白激素的第一个超活性类似物，其显著增加受体结合亲和力以及体外和体内生物效价和最大功效。在我们成功完成的2期SBIR研究期间，我们已经实现或超过了所有目标，包括筛选其他FSH类似物候选物，并确定用于临床试验的最佳FSH类似物TR 4401，使用多种体外和体内啮齿动物生物测定模型，包括经典的卵巢重量反应以及胚胎移植产生的卵母细胞，囊胚和新生幼崽。在所有这些啮齿动物模型中，4401在与定量终点以及与卵母细胞或胚胎质量相关的定性终点相关的效价和有效性方面均大大优于所有目前可用的重组FSH制剂。此外，我们使用模拟人类辅助生殖技术的方法，在代表两种人类不育模型的FSH受体表达降低的人类细胞系中，在两种不育的牛模型中和在恒河猴中，显示了TR 4401相对于标准FSH的这种上级功效。我们还表明，在奶牛中注射一次TR 4401可以产生与猪FSH的标准8次注射方案相当的超数排卵，重复给药后反应没有减弱。这些非人灵长类动物和牛研究显示，在假定的最大剂量下，TR 4401没有卵巢过度刺激的证据，也没有任何免疫原性的证据，这是FDA关注的这种最低限度修饰的FSH类似物仅有的两种副作用，因此假定是安全的。使用HPLC验证的免疫测定，我们发现TR 4401在啮齿动物和猴中的药代动力学特性优于标准FSH，这显然是延迟吸收的结果。我们还开发了一种稳定的中国人卵巢（CHO）细胞系，可生产高水平的最终TR 4401类似物;优化了大规模生物反应器生产方法;开发了适用于商业规模的新型高容量纯化方法;通过多种理化方法（包括碳水化合物分析）对纯化的类似物进行了严格的定量和表征。在本申请中，根据我们与FDA非常成功的IND前会议中获得的FDA的具体指令，我们提出了FDA要求的所有步骤，包括详细甘特中指出的具体时间表，以进一步商业开发这种新型FSH类似物。这些包括主CHO细胞库和工作CHO细胞库的建立和表征;另外两个大批次TR 4401的生产（每种>200 mg）：第一次在GLP下，第二次在GMP符合条件下;进行所有FDA要求的功效、特异性、稳定性、代谢、药代动力学、药效学和分析研究;执行所有FDA要求的毒理学研究，包括大鼠和家兔的两代生殖毒理学评估;以及提交IND以启动临床试验。我们已将TR 4401授权给全球领先的兽医超数排卵公司Bioniche，Inc。用于兽医用途，也可能选择共同开发用于人类用途的类似物。