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: 8195728
• 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/8195728
• 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; Screening procedure; 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; 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. PUBLIC HEALTH RELEVANCE: 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 shown in previous phase one and phase two SBIR grant studies that our novel FSH superagonist, TR4401, engineered to increase binding affinity to the FSH receptor, shows greatly increased potency and efficacy compared to currently available FSH products, in multiple quantitative and qualitative oocyte endpoints in rodents, monkeys, and bovines, with no apparent side effects. We now propose to commercialize this novel FSH analog by following all the specific FDA guidelines given to us in a pre-IND meeting in order to submit an IND to that agency to initiate clinical trials.

描述（由申请人提供）：不孕不育症影响着大约 10% 的美国夫妇，该领域的治疗市场非常大且增长迅速，特别是负责卵巢卵母细胞发育的主要激素，人卵泡刺激素 (FSH)。尽管现有的尿和重组 hFSH 产品已相当成功，但目前对于所有不孕女性，特别是老年患者和对当前疗法相对无反应的患者，对改进的 FSH 类似物的治疗需求尚未得到满足。我们之前已经描述了第一个超活性糖蛋白激素类似物，可显着增加受体结合亲和力以及体外和体内生物效力和最大功效。在我们成功完成的 2 期 SBIR 研究中，我们已经实现或超越了所有目标，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、囊胚和新生幼崽的反应。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力和功效方面均大大优于目前所有可用的重组 FSH 制剂。此外，我们使用模拟人类辅助生殖技术的方法，在代表两种人类不孕模型（两种牛不孕模型和恒河猴）的 FSH 受体表达降低的人类细胞系中显示出 TR4401 优于标准 FSH 的功效。我们还表明，在奶牛中注射一次 TR4401 可以产生与猪 FSH 标准 8 次注射方案相当的超数排卵，并且重复给药后反应不会减弱。这些非人灵长类动物和牛研究显示，没有证据表明TR4401在可能的最大剂量下会过度刺激卵巢，也没有任何免疫原性的证据，而免疫原性是FDA对这种经过最低限度修改、因此可能是安全的FSH类似物所关注的唯一两个副作用。使用经 HPLC 验证的免疫测定，我们发现 TR4401 在啮齿动物和猴子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。我们还开发了稳定的中国仓鼠卵巢 (CHO) 细胞系，可产生高水平的最终 TR4401 类似物；大规模生物反应器生产方法的优化；开发适合商业规模扩大的新型高容量纯化方法；通过多种物理化学方法（包括碳水化合物分析）对纯化类似物进行严格的定量和表征。在当前的申请中，根据 FDA 在我们与他们的非常成功的预 IND 会议上获得的具体指令，我们提出了 FDA 进一步商业开发这种新型 FSH 类似物所需的所有步骤，包括详细甘特图上指示的具体时间表。其中包括主和工作 CHO 细胞库的建立和表征；额外生产两批大批量 TR4401（每批 >200 毫克）：第一批在 GLP 条件下，第二批在 GMP 合规条件下；执行 FDA 要求的所有功效、特异性、稳定性、代谢、药代动力学、药效学和分析研究；执行 FDA 要求的所有毒理学研究，包括大鼠和兔子的两代生殖毒理学评估；并提交 IND 以启动临床试验。我们已将 TR4401 授权给全球领先的兽医超排卵公司 Bioniche, Inc.，用于兽医用途，也可以选择共同开发人类使用的类似物。 公共健康相关性：不孕不育症影响着大约 10% 的美国夫妇，该领域的治疗市场非常大且增长迅速，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)。尽管现有的尿和重组 hFSH 产品已相当成功，但目前对于所有不孕女性，特别是老年患者和对当前疗法相对无反应的患者，对改进的 FSH 类似物的治疗需求尚未得到满足。我们在之前的第一阶段和第二阶段 SBIR 资助研究中表明，我们的新型 FSH 超级激动剂 TR4401 经过精心设计，旨在增加与 FSH 受体的结合亲和力，与目前可用的 FSH 产品相比，在啮齿动物、猴子和牛的多个定量和定性卵母细胞终点中显示出大大提高的效力和功效，且没有明显的副作用。我们现在建议遵循 FDA 在 IND 前会议上向我们提供的所有具体指南，将这种新型 FSH 类似物商业化，以便向该机构提交 IND 来启动临床试验。