An In Vitro Robotic Assay for Anti-Estrogenic Activity

描述(申请人提供)：最近的科学研究表明，塑料、药品、杀虫剂、化妆品、食品添加剂等中使用的许多化学品都是内分泌干扰物(EDs)。EDS以不同的方式干扰雌激素等激素，对许多行为和生理过程产生显著的不利影响。ED效应(例如，雌激素或抗雌激素)有时在很低的浓度(皮摩尔到纳摩尔)下发生，特别是在胎儿或发育中的哺乳动物(包括人类)。EDS在我们环境中的流行和作用要求开发有效的检测方法。因此，各种政府机构(例如，EPA、FDA和ICCVAM)和积极主动的公司都明确表示希望对ED进行体外机器人检测，例如抗雌激素活性(anti-EA)。然而，目前还没有商业化的抗EA检测方法。为了开始满足这些政府、科学和商业需求，认证化学(CCI)拥有的初步数据表明，使用MCF-7细胞开发一种有效的(即可靠、准确、通用、快速和经济有效的)抗EA机器人检测是可行的。在这个第一阶段的应用中，CCI现在建议通过首先确定使用MCF-7细胞检测抗EA的最佳机器人方案来确认开发用于抗EA的机器人检测是可行的。CCI然后建议通过在12周内重复(5次)检测ICCVAM发布的用于验证抗EA检测(特定目标1)的有限参考化学物质(10)来评估该检测的可靠性(重复性)。CCI还建议(具体目标2)通过将CCI获得的这10种参考化学品的数据与ICCVAM公布的数据进行比较来评估这种分析的准确性。我们强烈期待在第一阶段获得的数据将证实我们的初步数据，即这种抗EA试验值得在第二阶段研究中进行更广泛的测试和开发，为其商业化做准备。开发一种用于抗EA的机器人筛查方法具有重要的商业、科学和社会意义，因为现在有大量的化学物质(&gt；10,000)--更不用说化学混合物了--现在应该由营利、非营利或政府实体对抗EA进行筛查。作为NIEHS、EPA和ICCVAM任务的一部分，希望开发一种机器人抗EA检测方法。

DESCRIPTION (provided by applicant): Recent scientific investigations have shown that many chemicals used in plastics, Pharmaceuticals, pesticides, cosmetics, food additives, etc., are endocrine disrupters (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 anti-EA assay is commercially available. To begin to meet these governmental, scientific and commercial needs, CertiChem (CCi) has preliminary data suggesting 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 I application, CCi now proposes to confirm that it is feasible to develop a robotic assay for anti-EA by first determining an optimum robotic protocol for detecting anti-EA using MCF-7 cells. CCi then proposes to estimate the reliability (reproducibility) of this assay by repeatedly (5 times) assaying over a 12 week period a limited set of reference chemicals (10) published by ICCVAM to use for validation of anti-EA assays (Specific Aim #1). CCi also proposes (Specific Aim #2) to estimate the accuracy of this assay by comparing data CCi obtains on these 10 reference chemicals with data published by ICCVAM. We strongly expect data obtained on this Phase I grant to confirm our preliminary data that this anti-EA assay is worthy of more extensive testing and development in a Phase II study to prepare it for commercialization. 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.