Identifying Small Molecules that Regulate Uterine Contractions

识别调节子宫收缩的小分子

基本信息

批准号：
9317253
负责人：
Jennifer L. Herington
金额：
$ 27.65万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-04-01 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9317253
关键词：
Adult Adverse effects Affinity Agonist Aorta Biological Assay Blood Vessels Cells Cesarean section Cessation of life Clinical Collection Detection Development Dimethyl Sulfoxide Discipline of obstetrics Dose Ductus Arteriosus Effectiveness Evaluation FDA approved Future Goals Human Infant Mortality Isometric Exercise Lead Libraries 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 inhibitor/antagonist myometrium novel preclinical development pregnant public health relevance response screening small molecule success therapeutic target uterine contractility

项目摘要

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）劳动指示和3）控制产后出血（PPH）。宫缩和用于PPH的子宫术受到其不良脱靶的限制和短期持续时间此外，有益于PTL，PPH的药物开发几乎缺乏迹象。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，选择性和效能的小分子。子宫收缩中的Ca2+ - 动物化和催产素（OT）tracelular Ca2+ - 的有效作用释放，在不存在的情况下从子宫肌层（UT-MYO）细胞中剥削细胞内Ca2+ - 释放和或或或或或或或或或或或或或或或或或或或或od或或或或或或或或或或或或od或或或或或或或或或或或obs OT的存在提供了一个出色的策略，可以分别发现新的子宫毒剂和/或毒液。因此，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）。（z´= 0.73）和DMSO耐受性。和II集合证明：1）出色的测定性能，2）使用鼠标UT-MYO的可行性 HTS的细胞3）鉴定出用于调节子宫内子宫能力的IMEDIATE测试的化合物收缩。该应用的目的是采用我们的HTS准备识别以识别以识别调节的小分子子宫肌层Ca2+型 - 具有高亲和力和大型化合物的选择性我们将：1）在Selleckchem FDA-中扣押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Ca2+液体化合物在UT-MYO细胞中；经过比较屏幕和响应关系的“命中”确认和评估后 AIM 2中的分析我们将使用离体功能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。我们的研究的成功结合将确定一系列临床前的新铅化合物子宫肌层收缩的选择性调节剂的发展。