Development of Superagonist Analogs of Recombinant Human TSH for Imaging and Ther

用于成像和治疗的重组人 TSH 超级激动剂类似物的开发

• 批准号: 7673335
• 负责人: Bruce Dale Weintraub
• 金额: $ 88.02万
• 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 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7673335
• 关键词: Ablation; Affinity; Beds; Bioreactors; Cancer Patient; Data; Detection; Development; Diagnostic; Diagnostic Imaging; Dose; Early Diagnosis; Endocrine; Generations; Glycoproteins; Grant; High Pressure Liquid Chromatography; Hormones; Human; I125 isotope; Image; Immunoassay; In Vitro; Intramuscular; Intravenous; Iodine; Lead; Malignant Neoplasms; Malignant neoplasm of thyroid; Measurement; Metabolic Clearance Rate; Methods; Modeling; Mus; Organ; Papillary thyroid carcinoma; Patients; Phase; Plasma; Positron-Emission Tomography; Production; Protein Kinase Inhibitors; Radio; Recombinants; Recurrent tumor; Reporting; Research Personnel; Rodent; Small Business Innovation Research Grant; Staging; Structure; Technology; Therapeutic; Therapeutic Effect; Thyroglobulin; Thyroid Gland; Thyrotropin; Time; Tissues; Transfection; Transgenic Mice; Transgenic Organisms; analog; cancer imaging; cancer therapy; clinically relevant; commercialization; comparative efficacy; expression vector; fluorodeoxyglucose; hormone analog; improved; in vitro Assay; in vivo; in vivo Bioassay; intraperitoneal; method development; mouse model; novel; protein kinase inhibitor; public health relevance; receptor binding; response; scale up; stable cell line; subcutaneous; thyroid neoplasm; tumor; uptake

DESCRIPTION (provided by applicant): Thyroid cancer is increasingly prevalent and 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 thyroid remnant or tumor ablation with radioiodine therapy. The PI, who was both co-inventor and co-developer with Genzyme of wild type recombinant human (rh) TSH (Thyrogen) currently approved for stimulation of diagnostic radioiodine uptake and thyroglobulin secretion, now proposes to develop a more potent and more efficacious second generation superactive analog of rh TSH for the 15-20% of poorly responsive thyroid cancer patients in diagnostic imaging as well as for the majority of such patients for radioiodine therapy. 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. During our highly scored and successfully completed phase 1 SBIR study we have achieved all the aims including developing stable cell lines producing high levels of the final two candidate superactive analogs; optimization of large scale bioreactor production methods; development of a novel, high capacity purification methods suitable for commercial scale-up; rigorous quantification and characterization of purified analogs by multiple physicochemical methods, and proof of increased potency and efficacy in the now classic in vivo mouse rh TSH in vivo bioassay first developed by the PI. In the current phase 2 proposal, for which we have provided extensive preliminary data well beyond the aims of the phase 1 proposal, we plan to produce and purify additional large amounts (>200mg) of the final two TSH analog candidates TR1401 and 1402 in the fibrous bed bioreactor sufficient for all the phase two in vivo studies using the methods of production and purification optimized in the phase one study. We also plan to develop and validate a novel immunoassay for the detection of TSH analogs in unpurified rodent plasma that quantifies accurately and identically to rigorous HPLC methods which will then be used to determine the IV metabolic clearance rate and the IP, SC and IM phamacokinetics in rodents. Most importantly, in this phase two proposal we will examine for the first time multiple clinically relevant diagnostic and therapeutic endpoints of TSH action comparing TSH analogs TR1401 and 1402 to wild type (Genzyme) recombinant TSH in normal mice and in those with Ret/PTC1 and other transgenic mouse models of thyroid cancer. Specifically we will examine detailed dose response curves for both wild type and analog TSH in thyroidal 125I uptake by direct organ counting, diagnostic thyroidal PET imaging with 124I and 18 Fluorodeoxyglucose as well as therapeutic 131I -induced tumor regression. Finally, we will determine the possible complementation of the diagnostic and therapeutic effects of TR1401 and 1402 with those of selected newly developed protein kinase inhibitors in vitro and in vivo. PUBLIC HEALTH RELEVANCE: Thyroid cancer is the most common malignancy of endocrine tissues and most patients require lifelong surveillance with diagnostic thyroid-stimulating hormone (TSH)-stimulated radio-iodine imaging to detect recurrent tumor. The PI and Co- Investigator have developed a much improved form of TSH and propose a completely new method of thyroid cancer imaging that should greatly improve early detection and lead to improved cancer treatment and survival.

描述（由申请人提供）：甲状腺癌越来越普遍，是内分泌组织最常见的恶性肿瘤，大多数患者需要通过诊断性促甲状腺激素（TSH）刺激的放射性碘摄取进行终生监测，以及通过放射性碘治疗进行甲状腺残余或肿瘤消融。该 PI 是 Genzyme 野生型重组人 (rh) TSH (Thyrogen) 的共同发明者和共同开发者，目前已批准用于刺激诊断性放射性碘摄取和甲状腺球蛋白分泌，现在提议开发一种更有效、更有效的第二代 rh TSH 超活性类似物，用于诊断成像中 15-20% 反应不佳的甲状腺癌患者： 以及对大多数此类患者进行放射性碘治疗。我们之前的结构功能研究发现了 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显着增加。在我们高分并成功完成的 1 期 SBIR 研究中，我们实现了所有目标，包括开发稳定的细胞系，生产高水平的最后两种候选超活性类似物；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对纯化类似物进行严格的定量和表征，并证明 PI 首次开发的现在经典的体内小鼠 rh TSH 体内生物测定法的效力和功效有所增加。在当前的第 2 阶段提案中，我们提供了远远超出第 1 阶段提案目标的广泛初步数据，我们计划使用在第 1 阶段研究中优化的生产和纯化方法，在纤维床生物反应器中生产和纯化额外大量（>200mg）的最后两种 TSH 类似物候选物 TR1401 和 1402，足以用于所有第 2 阶段体内研究。我们还计划开发和验证一种新型免疫测定法，用于检测未纯化的啮齿动物血浆中的 TSH 类似物，其定量准确且与严格的 HPLC 方法相同，然后用于确定啮齿动物的 IV 代谢清除率以及 IP、SC 和 IM 药代动力学。最重要的是，在这个第二阶段提案中，我们将首次检查 TSH 作用的多个临床相关诊断和治疗终点，将 TSH 类似物 TR1401 和 1402 与正常小鼠、Ret/PTC1 和其他甲状腺癌转基因小鼠模型中的野生型 (Genzyme) 重组 TSH 进行比较。具体来说，我们将通过直接器官计数、用 124I 和 18 氟脱氧葡萄糖进行诊断性甲状腺 PET 成像以及治疗性 131I 诱导的肿瘤消退来检查甲状腺 125I 摄取中野生型和模拟 TSH 的详细剂量反应曲线。最后，我们将确定 TR1401 和 1402 与选定的新开发的蛋白激酶抑制剂在体外和体内的诊断和治疗效果的可能互补。公共健康相关性：甲状腺癌是内分泌组织最常见的恶性肿瘤，大多数患者需要通过诊断性促甲状腺激素 (TSH) 刺激的放射性碘成像进行终生监测，以检测复发性肿瘤。 PI 和合作研究者开发了一种大大改进的 TSH 形式，并提出了一种全新的甲状腺癌成像方法，该方法将大大改善早期检测并改善癌症治疗和生存。