LONG ACTING GONADOTROPIN

长效促性腺激素

• 批准号: 2749575
• 负责人: WILLIAM R MOYLE
• 金额: $ 21.35万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-15 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2749575
• 关键词: CHO cells; SDS polyacrylamide gel electrophoresis; chemical substitution; chorionic gonadotropin; clearance rate; disulfide bond; drug design /synthesis /production; drug screening /evaluation; electrofocusing; follicle stimulating hormone; gonadotropins; hormone analog; hormone binding protein; hormone metabolism; hormone receptor; laboratory rat; luteinizing hormone; ovulation; pharmacokinetics; radiotracer; receptor binding; site directed mutagenesis; spermatogenesis

Recent progress in our understanding of the structure and function of gonadotropins has led to the development of hormone analogs that can be tailored to have desirable levels of LH and/or FSH activities. It is now possible to design analogs of human chorionic gonadotropin (hCG) that retain virtually the entire structure of hCG but that function more like FSH. Indeed, some are even more potent than FSH in vivo. Other analogs of hCG have been developed that bind to both LH and FSH receptors. The abilities of these analogs to mimic LH or FSH in vivo are unknown. Studies proposed in Aim 1 are designed to develop a long-acting bifunctional gonadotropin agonist. This will be accomplished in part by attaching the C-terminus of the hCG Beta-subunit to an analog known to have dual LH and FSH activity. Studies in Aim 1 will also determine if creating additional glycosylation signals in the alpha and Beta-subunits will prolong the half-life of this hCG analog. The studies in Aims 2 and 3 are a prelude to the development of a potent bi-functional LH/FSH antagonist that will have a long plasma half-life. In Aim 2 we will determine the effects of removing the N- and O-linked oligosaccharides from hCG on its clearance. While it is well known that chemical or enzymatic deglycosylation reduces the half-life of gonadotropins, the effects of genetic deglycosylation are unknown. In Aim 3 we will determine if specific amino acids in the Beta-subunits of hCG, FSH, and bovine LH are responsible for their different turnover rates. In Aim 3 we will also study the effects of inserting an inter-subunit disulfide bond on hormone activity and clearance. The hypothesis to be tested is that gonadotropin clearance is partly a consequence of subunit dissociation. The presence of an inter-subunit disulfide may slow subunit dissociation and substantially reduce clearance. In Aim 4 we will prepare and test a deglycosylated analog of hCG that binds to both LH and FSH receptors to learn if it blocks the actions of both LH and FSH. The final design of this analog will depend on what is learned about prolonging the half-lives of deglycosylated gonadotropins in Aims 2 and 3.

我们对结构和功能理解的最新进展 促性腺激素导致了激素类似物的开发 旨在获得理想的 LH 和/或 FSH 活性水平。现在是 可以设计人绒毛膜促性腺激素（hCG）的类似物 实际上保留了 hCG 的整个结构，但其功能更像 卵泡刺激素。事实上，有些在体内甚至比 FSH 更有效。其他类似物 已开发出与 LH 和 FSH 受体结合的 hCG。这 这些类似物在体内模拟 LH 或 FSH 的能力尚不清楚。 目标 1 中提出的研究旨在开发一种长效 双功能促性腺激素激动剂。这将部分完成 将 hCG β 亚基的 C 末端连接到已知的类似物上 具有双重 LH 和 FSH 活性。目标 1 的研究还将确定是否 在 α 和 Beta 亚基中产生额外的糖基化信号 将延长这种 hCG 类似物的半衰期。目标 2 和 3 是开发强效双功能 LH/FSH 的前奏 具有长血浆半衰期的拮抗剂。在目标 2 中，我们将 确定去除 N- 和 O- 连接寡糖的效果 来自 hCG 的清除。虽然众所周知，化学或 酶促去糖基化会缩短促性腺激素的半衰期， 遗传去糖基化的影响尚不清楚。在目标 3 中，我们将 确定 hCG、FSH 和 β 亚基中的特定氨基酸是否 牛 LH 的周转率不同。目标 3 我们还将研究插入亚基间二硫键的影响 与激素活性和清除率有关。要检验的假设是 促性腺激素清除部分是亚基的结果 解离。亚基间二硫键的存在可能会减慢亚基的速度 解离并显着降低清除率。在目标 4 中，我们将准备 并测试与 LH 和 FSH 结合的 hCG 去糖基类似物 受体以了解它是否阻断 LH 和 FSH 的作用。决赛 这个模拟的设计将取决于关于延长时间的知识 目标 2 和 3 中去糖基化促性腺激素的半衰期。