Development of Superagonist Analogs of Recombinant Human TSH

重组人 TSH 超级激动剂类似物的开发

• 批准号: 7108446
• 负责人: Bruce Dale Weintraub
• 金额: $ 10.15万
• 依托单位: TROPHOGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-15 至 2006-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7108446
• 关键词: CHO cells; analytical ultracentrifugation; bioassay; biotechnology; dosage; drug design /synthesis /production; hormone analog; laboratory mouse; neoplasm /cancer chemotherapy; protein purification; recombinant proteins; stimulant /agonist; thyroid neoplasm; thyrotropin; transfection /expression vector

DESCRIPTION (provided by applicant): Thyroid cancer is the most common malignancy of endocrine tissues and most patients require lifelong surveillance with diagnostic thyroid-stimulating hormone (TSH)-stimulated radioiodine uptake as well as remnant or metastatic ablation with radioidine therapy. The PI, who was both co- inventor and co-developer with Genzyme of wild-type recombinant human (rh) TSH currently approved for diagnosis, now proposes to develop more potent and more efficacious second generation analogs of rh TSH for the 10-20% of poorly responsive patients. Our previous structure- function studies resulted in the discovery of the first analogs of TSH and other glycoprotein hormones with major increases in receptor binding affinity and bioactivity ("superactive analogs"). In the current proposal we plan to produce, purify and characterize large amounts of such novel TSH analogs for commercial development. Since TSH in vivo bioassays require higher quantities of hormones than that obtainable from transient transfection experiments, we propose in Aim 1 to develop stable cell lines producing high levels of candidate superactive analogs and produce quantities sufficient for initial animal studies. This Aim will consist of construction of dicistronic vectors with amplifiable markers and mutated subunit cDNAs, amplification, production and concentration of unpurified analogs. Aim 2 will consist of development of a novel, high capacity analog purification method suitable for commercial scale-up. Aim 3 will consist of rigorous quantification and characterization of selected purified analogs by multiple physicochemical methods, including carbohydrate analysis. In Aim 4 analogs will be examined in the now classic mouse rh TSH in vivo bioassay first developed by the PI, with full dose response curves to assess increased potency and maximal efficacy of TSH analogs in order to select a lead candidate for further commercial development.

描述（由申请方提供）：甲状腺癌是最常见的内分泌组织恶性肿瘤，大多数患者需要终身监测，诊断性促甲状腺激素（TSH）刺激的放射性碘摄取以及放射性碘治疗的残留或转移消融。PI是Genzyme目前批准用于诊断的野生型重组人（rh）TSH的共同发明者和共同开发者，现在提议开发更强效和更有效的第二代rh TSH类似物，用于10-20%的反应不良患者。我们先前的结构-功能研究导致了TSH和其他糖蛋白激素的第一个类似物的发现，这些类似物在受体结合亲和力和生物活性方面具有显著增加（“超活性类似物”）。在目前的提案中，我们计划生产，纯化和表征大量的这种新型TSH类似物用于商业开发。由于促甲状腺激素在体内的生物测定需要更高数量的激素比从瞬时转染实验中获得的，我们建议在目标1开发稳定的细胞系产生高水平的候选超活性类似物，并产生足够的数量为初始的动物研究。该目标将包括构建具有可扩增标记和突变的亚基cDNA的双顺反子载体，扩增、产生和浓缩未纯化的类似物。目标2将包括开发一种新的，高容量的类似物纯化方法，适用于商业规模。目标3将包括通过多种物理化学方法（包括碳水化合物分析）对选定的纯化类似物进行严格的定量和表征。在目的4中，将在PI首次开发的现在经典的小鼠rh TSH体内生物测定中检查类似物，使用完整的剂量反应曲线来评估TSH类似物的增加的效力和最大功效，以选择用于进一步商业开发的主要候选物。