Department of Molecular Medicine, The Scripps Research Molecular Screening Center, Scripps Research Florida, 130 Scripps Way #1A1, Jupiter, FL, 33458, USA.
Texas Tech University Health Sciences Center, 3601 4th Street, Lubbock, TX, USA.
Sci Rep. 2020 Jun 29;10(1):10579. doi: 10.1038/s41598-020-67473-w.
Molecules that correct the folding of protein mutants, restoring their functional trafficking, are called pharmacoperones. Most are clinically irrelevant and possess intrinsic antagonist or agonist activity. Here, we identify compounds capable of rescuing the activity of mutant gonadotropin-releasing hormone receptor or GnRHR which, is sequestered within the cell and if dysfunctional leads to Hypogonadotropic Hypogonadism. To do this we screened the E90K GnRHR mutant vs. a library of 645,000 compounds using a cell-based calcium detection system. Ultimately, we identified 399 compounds with EC ≤ 5 µM with no effect in counterscreen assays. Medicinal chemistry efforts confirmed activity of 70 pure samples and mode of action studies, including radioligand binding, inositol phosphate, and toxicity assays, proved that we have a series of tractable compounds that can be categorized into structural clusters. These early lead molecules rescue mutant GnRHR function and are neither agonist nor antagonists of the GnRHR cognate receptor, a feature required for potential clinical utility.
能够纠正蛋白质突变体折叠，恢复其功能运输的分子被称为药效协同分子。大多数都与临床无关，具有内在的拮抗剂或激动剂活性。在这里，我们鉴定出能够恢复突变型促性腺激素释放激素受体（GnRHR）活性的化合物，该受体被隔离在细胞内，如果功能失调会导致促性腺激素释放激素缺乏性性腺功能减退症。为此，我们使用基于细胞的钙检测系统筛选了 E90K GnRHR 突变体与 645000 种化合物库。最终，我们鉴定出 399 种 EC≤5µM 的化合物，在对照筛选试验中没有效果。药物化学研究证实了 70 种纯样品的活性和作用机制研究，包括放射性配体结合、三磷酸肌醇和毒性测定，证明我们有一系列可处理的化合物，可以归类为结构簇。这些早期的先导分子可以挽救突变型 GnRHR 的功能，既不是 GnRHR 同源受体的激动剂也不是拮抗剂，这是潜在临床应用所必需的特征。
