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（由申请人提供）：甲状腺癌是内分泌组织最常见的恶性肿瘤，大多数患者需要终身监测诊断性甲状腺刺激激素（TSH）刺激的放射性碘摄取以及用于放射性疗法的残留或转移性消融。 PI既是野生型重组人（RH）TSH的共同发明者又是共同开发者，目前已批准进行诊断，现在提议为10-20％的不良反应性患者开发更有效，更有效的RH TSH的第二代类似物。我们先前的结构功能研究导致发现了TSH和其他糖蛋白激素的第一个类似物，其受体结合亲和力和生物活性的大大增加（“超级活性类似物”）。在当前的建议中，我们计划生产，净化和表征大量此类新型TSH类似物用于商业开发。由于TSH在体内生物测定需要比瞬态转染实验获得的激素量更高，因此我们在AIM 1中提出，以开发稳定的细胞系，产生高水平的候选超活性类似物，并产生足以初始动物研究的数量。这个目标将包括具有放大标记和突变的亚基cDNA，扩增，生产和浓度未纯化的类似物的二氧化载体载体的构建。 AIM 2将包括一种适用于商业规模的新型高容量模拟纯化方法的开发。 AIM 3将包括通过多种物理化学方法（包括碳水化合物分析）对所选纯化类似物的严格定量和表征。在AIM 4中，将在现在由PI首次开发的现有经典的小鼠RH TSH中进行研究，并具有全剂量响应曲线，以评估TSH类似物的效力和最大功效，以便为进一步的商业开发选择主要候选者。