Identifying Small Molecules that Regulate Uterine Contractions

识别调节子宫收缩的小分子

• 批准号: 9444468
• 负责人: Jennifer L. Herington
• 金额: $ 15.8万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9444468
• 关键词: Adult; Adverse effects; Affinity; Agonist; Aorta; Biological Assay; Blood Vessels; Cells; Cesarean section; Clinical; Collection; Detection; Development; Dimethyl Sulfoxide; Discipline of obstetrics; Dose; Ductus Arteriosus; Effectiveness; Evaluation; FDA approved; Future; Goals; Human; Induced Labor; Isometric Exercise; Lead; Libraries; Maternal Mortality; Methods; Modeling; Molecular; Mus; Myometrial; Organ; Outcome; Oxytocin; Pathway interactions; Performance; Pharmaceutical Preparations; Postpartum Hemorrhage; Pregnancy; Premature Birth; Premature Labor; PubChem; Publishing; Regulation; Reproducibility; Research; Research Personnel; Role; Safety; Series; Signal Pathway; Smooth Muscle Myocytes; Techniques; Testing; Therapeutic; Therapeutic Effect; Time; Tissues; Tocolytic Agents; United States National Institutes of Health; Uterine Contraction; Uterine Inertia; Woman; base; comparative; drug development; drug discovery; fetal; high throughput screening; improved; infant morbidity/mortality; inhibitor/antagonist; myometrium; novel; preclinical development; pregnant; public health relevance; response; screening; small molecule; success; therapeutic target; uterine contractility

PROJECT SUMMARY The uterine myometrium is a primary therapeutic target for: 1) mitigation of preterm labor (PTL), 2) labor induction, and 3) control of postpartum hemorrhage (PPH). Current tocolytics used to inhibit uterine contractions, and uterotonics used for PPH, are limited by their undesirable off-target effects and short duration of benefit. Moreover, there is almost a complete lack of drug development for PTL, PPH and other obstetric indications. Thus, a substantial need exists for novel, safe tocolytic and uterotonic agents with improved efficacy and selectivity. High-throughput screening (HTS) provides a drug-discovery platform for researchers to identify and then optimize small-molecules with increased affinity, selectivity and efficacy/potency. Based on the central role of Ca2+-mobilization in uterine contractions, and potent effects of oxytocin (OT) on uterine intracellular Ca2+- release, exploitation of intracellular Ca2+-release from uterine myometrial (UT-myo) cells in the absence and/or presence of OT provides an excellent strategy to discover new uterotonics and/or tocolytics, respectively. Thus, we developed a dual-addition Ca2+-mobilization assay, using a fluorescent Ca2+-sensitive probe and primary mouse UT-Myo cells, to allow dual-detection of agonists (uterotonics) of Ca2+-mobilization and antagonists (tocolytics) of OT-induced Ca2+-mobilization in a single screen. This assay is robust, reproducible (Z´ =0.73), and DMSO tolerant. A pilot screen of 2,727 compounds from the Spectrum Collection, NIH Clinical I and II Collections demonstrated: 1) excellent assay performance, 2) feasibility of using primary mouse UT-myo cells for HTS and 3) identified compounds for immediate testing for ability to regulate ex vivo uterine contractions. The goal of this application is to employ our HTS-ready assay to identify small molecules that regulate uterine myometrial Ca2+-mobilization with high affinity and selectivity from a large-compound library. In Aim 1 we will: 1) implement a large-scale HTS-campaign against 100,978 compounds in the SelleckChem FDA- approved Drug Library and Vanderbilt Discovery Collection, to identify hit-compounds that stimulate Ca2+- mobilization or inhibit OT-induced Ca2+-mobilization in UT-myo cells; and 2) narrow hits to lead-compounds after confirmation and evaluation of "hits" undergoing a comparative screen and dose-response relationship analysis. In Aim 2 we will validate lead small-molecules by using an ex vivo functional isometric contractile assay to record the therapeutic-effects of lead-compounds on human uterine myometrial contractile activity. The successful completion of our studies will identify a series of new lead-compounds for pre-clinical development of selective regulators of uterine myometrial contractility.

项目摘要 子宫肌层是主要的治疗目标：1）减轻早产（PTL），2）分娩 3）控制产后出血（PPH）。目前用于抑制子宫收缩的宫缩抑制剂 PPH中使用的宫缩和子宫收缩剂受到其不良脱靶效应和持续时间短的限制 利益。此外，几乎完全缺乏用于PTL、PPH和其他产科疾病的药物开发。 迹象。因此，存在对具有改善的促排卵活性的新型、安全的宫缩抑制剂和子宫收缩剂的实质性需求。 有效性和选择性。 高通量筛选（HTS）为研究人员提供了一个药物发现平台， 优化小分子，提高亲和力、选择性和功效/效力。基于核心作用， 子宫收缩中的Ca ~（2+）动员和催产素（OT）对子宫细胞内Ca ~（2+）的有效作用 释放，利用子宫肌层（UT-myo）细胞在不存在和/或 OT的存在分别为发现新的子宫收缩剂和/或宫缩抑制剂提供了极好的策略。 因此，我们开发了一种双添加Ca 2+动员试验，使用荧光Ca 2+敏感探针， 原代小鼠UT-Myo细胞，以允许双重检测Ca 2+动员的激动剂（子宫收缩剂）和 OT诱导的Ca 2+动员的拮抗剂（宫缩抑制剂）。该试验具有耐用性、重现性 （Z ′ =0.73），耐DMSO。对来自光谱收集，NIH临床I的2，727种化合物进行了初步筛选 和II收集证明：1）优异的测定性能，2）使用原代小鼠UT-myo 细胞进行HTS和3）鉴定化合物用于立即测试调节离体子宫内膜的能力， 宫缩 本申请的目标是采用我们的HTS-就绪测定来鉴定调节细胞凋亡的小分子。 子宫肌层Ca 2+动员具有高亲和力和选择性，来自大型化合物库。目标1 我们将：1）对SelleckChem FDA中的100，978种化合物实施大规模HTS活动- 批准的药物库和范德比尔特发现收藏，以确定刺激钙离子的命中化合物， UT-myo细胞中OT诱导的Ca 2+动员或抑制OT诱导的Ca 2+动员;和2）缩小对铅化合物的命中 在确认和评价“命中”后，进行比较筛选和剂量-反应关系 分析.在目标2中，我们将通过使用离体功能性等长收缩剂来验证先导小分子 记录铅化合物对人子宫肌层收缩活性的治疗效果的测定。 我们的研究的成功完成将确定一系列新的先导化合物用于临床前研究。 子宫肌层收缩力的选择性调节剂的开发。

项目成果

Effects of Solvents, Emulsions, Cosolvents, and Complexions on Ex Vivo Mouse Myometrial Contractility.

• DOI: 10.1007/s43032-021-00576-5
• 发表时间: 2022-03
• 期刊: Reproductive sciences (Thousand Oaks, Calif.)
• 作者: Hansen CJ;Siricilla S;Boatwright N;Rogers JH;Kumi ME;Herington J
• 通讯作者: Herington J

Drug discovery strategies for the identification of novel regulators of uterine contractility.

用于识别新型子宫收缩调节剂的药物发现策略。

• DOI: 10.1016/j.cophys.2019.10.012
• 发表时间: 2020
• 期刊: Current opinion in physiology
• 影响因子: 2.5
• 作者: Siricilla,Shajila;Iwueke,ChisomC;Herington,JenniferL
• 通讯作者: Herington,JenniferL