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 子宫子宫骨术是：1）缓解早产（PTL）的主要治疗靶点，2）劳动 诱导和3）控制产后出血（PPH）。用于抑制子宫的电流溶剂剂 宫缩和用于PPH的子宫学，受其不受欢迎的脱靶效果和短持续时间的限制 利益。此外，PTL，PPH和其他产科几乎完全缺乏药物开发 适应症。这是对新颖，安全的溶溶和子宫毒剂的巨大需求，并有所改善 功效和选择性。 高通量筛查（HTS）为研究人员提供了一个药物发现平台，以识别然后 优化具有增加亲和力，选择性和效率/效力的小分子。基于中心角色 子宫收缩中的Ca2+ - 动物化，氧气（OT）对子宫内细胞内Ca2+ - 的潜在影响 在不存在和/或 OT的存在提供了一个出色的策略，可以分别发现新的子宫剂和/或糖尿病。 这，我们使用荧光Ca2+敏感探针和 初级小鼠UT-myo细胞，以允许Ca2+固定化的激动剂（子宫核）双重检测 OT诱导的Ca2+ - 动物在单个筛选中的拮抗剂（淋巴结剂）。该测定很强，可再现 （z´= 0.73）和DMSO耐受性。来自Spectrum Collection的2,727种化合物的试验屏幕，NIH临床I 和II集合所证明：1）出色的测定性能，2）使用主要鼠标UT-MYO的可行性 HTS的细胞和3）鉴定的化合物，用于立即测试调节子宫肾的能力 收缩。 该应用的目的是使用我们的HTS就绪测定法来识别调节的小分子 子宫肌层Ca2+摩擦型具有高亲和力和大型化合物库的选择性。在目标1中 我们将：1）在SelleckChem FDA-中实施大规模的HTS-Campaign，可针对100,978种化合物 批准的药品库和范德比尔特发现收集，以识别刺激Ca2+ - 动员或抑制OT诱导的UT-MYO细胞中的Ca2+润滑；和2）狭窄的铅化合物 经过比较屏幕和剂量反应关系的“命中”确认和评估后 分析。在AIM 2中，我们将通过使用离体功能等距收缩来验证铅小分子 测定以记录人子宫肌层收缩活性的铅化合物的治疗效应。 我们的研究的成功完成将确定一系列新的铅化合物，用于临床前 开发子宫肌层收缩性的选择性调节剂。

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