An In Vitro Robotic Assay for Anti-Estrogenic Activity

描述（由申请人提供）：最近的科学调查表明，许多用于塑料、药品、农药、化妆品、食品添加剂等的化学品，内分泌干扰物（EDs）。ED以各种方式干扰激素，如雌激素，对许多行为和生理过程产生显着的不利影响。艾德效应（例如雌激素或抗雌激素）有时在非常低的浓度（皮摩尔至纳摩尔）下发生，尤其是对胎儿或发育中的哺乳动物（包括人类）。ED在我们环境中的流行和作用保证了有效的检测方法的发展。因此，各种政府机构（例如，EPA、FDA和ICCVAM）和积极主动的公司已经明确表达了对ED进行体外机器人测定的愿望，例如抗雌激素活性（抗EA）。然而，没有机器人抗EA测定是商业上可获得的。为了开始满足这些政府、科学和商业需求，CertiChem（CCi）已经完成了第一阶段SBIR授权，表明开发有效的抗EA机器人测定是可行的（即，可靠、准确、通用、快速和成本效益）。在该第二阶段申请中，CCi现在提出通过重复测定一组78种参考测试化学品来开发用于商业化的抗EA的机器人测定，以确认CCi提出的抗EA测定的可靠性和准确性。CCi还建议通过检查食品和塑料中使用的一组抗氧化剂中的抗EA来证明该测定的多功能性。最后，CCi建议对一些参比供试化学品进行HPLC分离测定，以确认其抗EA不是由于污染引起的，并提高该测定法用于商业用途的多功能性。开发用于抗EA的机器人筛选测定在商业上、科学上和社会上是重要的，因为大量的化学品（> 10，000）-更少的化学混合物-现在应该由营利、非营利或政府实体筛选抗EA。作为NIEHS、EPA和ICCVAM的使命的一部分，需要开发用于抗EA的机器人测定。最近的科学调查表明，许多（可能超过10，000）用于塑料，农药，化妆品和食品添加剂等常见产品的化学品具有雌激素/抗雌激素活性，严重干扰正常的雌激素作用，对人类的许多行为，生殖和生理过程产生不利影响。在本II期提案中，CertiChem（CCi）建议开发一种非常灵敏和准确的检测方法，以测量一组参考测试化学品和常见消费品中的抗雌激素活性。CCi建议使用这些数据和CCi关于许多化学品的雌激素活性的数据，以确定哪些化学品具有抗雌激素活性，并与各种公司合作，设计不含释放抗雌激素或雌激素活性的化学品的消费品。

DESCRIPTION (provided by applicant): Recent scientific investigations have shown that many chemicals used in plastics, pharmaceuticals, pesticides, cosmetics, food additives, etc., are endocrine disruptors (EDs). EDs interfere in various ways with hormones such as estrogens to have significant adverse effects on many behavioral and physiological processes. ED effects (e.g. estrogenic or anti-estrogenic) sometimes occur at very low (picomolar to nanomolar) concentrations, especially on fetal or developing mammals (including humans). The prevalence and actions of EDs in our environment warrant the development of valid assay methods. Consequently, various governmental bodies (e.g., EPA, FDA, and ICCVAM) and proactive corporations have explicitly expressed a desire to have in vitro robotic assays for EDs, such as anti-estrogenic activity (anti-EA). However, no robotic anti-EA assay is commercially available. To begin to meet these governmental, scientific and commercial needs, CertiChem (CCi) has completed a Phase I SBIR grant showing that it is feasible to develop an anti- EA robotic assay that would be valid (i.e., reliable, accurate, versatile, rapid, and cost effective) using MCF-7 cells. In this Phase II application, CCi now proposes to develop this robotic assay for anti-EA for commercialization by repeatedly assaying a set of 78 reference test chemicals to confirm the reliability and accuracy of CCi's proposed anti- EA assay. CCi also proposes to demonstrate the versatility of this assay by examining the anti-EA in a set of antioxidants used in foodstuffs and plastics. Finally, CCi proposes to perform HPLC separation assays on some reference test chemicals to confirm that their anti-EA is not due to contamination and to enhance the versatility of this assay for commercial use. Development of a robotic screening assay for anti-EA is commercially, scientifically, and socially important because of the large number of chemicals (>10,000) - much less chemical mixtures - that should now be screened for anti-EA by profit, non-profit, or governmental entities. Development of a robotic assay for anti-EA is desired as part of the mission of NIEHS, EPA and ICCVAM. Recent scientific investigations have shown that many (perhaps over 10,000) chemicals used in common products such as plastics, pesticides, cosmetics, and food additives have estrogenic/anti-estrogenic activities that severely interfere with normal estrogen actions to produce adverse effects on many behavioral, reproductive, and physiological processes in humans. In this Phase II proposal, CertiChem (CCi) proposes to develop a very sensitive and accurate assay to measure anti-estrogenic activity in a set of reference test chemicals and in common consumer products. CCi proposes to use these data and CCi's data on estrogenic activity on many chemicals to identify which chemicals have anti-estrogenic activity and to work with various firms to design consumer products that are free of chemicals that release anti-estrogenic or estrogenic activities.