Evaluation of the potential of Ashwagandha extracts to produce CYP-mediated drug interactions.

项目摘要 尽管在新药开发过程中会定期评估药物间相互作用，但 很少评价植物药和植物膳食补充剂引起天然产物-药物相互作用。到 为了纠正这一缺陷，我们正在响应PAR-20-228：增加NIH影响的试点项目 植物和其他天然产品推进研究中心（PI 2 CARBON）我们特别 建议评估Ashwagandha（Withania somnifera），美国使用的40种最畅销的植物之一 消费者在2018年，为抑制和诱导最重要的I期药物代谢酶， 细胞色素P450（CYP）。迄今为止，关于以下物质的抑制作用的报道有限： Ashwagandha提取物对CYP的影响，并且没有发表的研究检查Ashwagandha提取物是否具有 诱导表达的潜力。Ashwagandha特别被老年人用来缓解压力， 失眠和认知缺陷;这些人群经常服用常规药物治疗其他疾病， 健康状况。因此，老年人使用Ashwagandha研究了Ashwagandha如何影响大脑活动， 表达了一个重要的关切。Ashwagandha是两种正在研究的植物之一， 位于俄勒冈州的BENFRA植物膳食补充剂研究中心（BDSRC）的健康和 科学大学（U19 AT 010829）。我们的工作将与BENFRA BDSRC的工作协同， 提供Ashwagandha根和叶的经认证和化学表征的提取物， 申请人在奥本大学的实验室。对于我们的项目，我们提出了两个具体目标。在目标1中， 我们将筛选不同类型的Ashwagandha提取物对9种人肝CYP的抑制作用（包括所有 FDA推荐用于药物-药物评价的CYP）。合并人肝脏中CYP的抑制 微粒体将使用UPLC-MS探针底物和我们的超快UHPLC-MS/MS鸡尾酒测定法进行测量。在 目的2，我们将评估Ashwagandha提取物引起的mRNA表达的诱导， 根据FDA体外药物相互作用指南，夹心培养中的人肝细胞 问题研究将提供Ashwagandha提取物-水相互作用的综合数据 通过科学出版物。将证明Ashwagandha对CYP的抑制或诱导 随后是未来的研究，旨在确定造成所观察到的影响的化合物。 在证明与辅酶A酶的相互作用后，将进行以下方面的确证性临床研究： Ashwagandha-药物药代动力学相互作用。

Project Summary Although drug-drug interactions are routinely evaluated during new drug development, the potential for botanicals and botanical dietary supplements to cause natural product-drug interactions is rarely evaluated. To correct this deficiency, we are responding to the PAR-20-228: Pilot Projects Increasing the Impact of the NIH Centers for Advancing Research on Botanicals and Other Natural Products (PI2 CARBON). Specifically, we propose to evaluate Ashwagandha (Withania somnifera), one of the top 40 selling botanicals used by US consumers in 2018, for inhibition and induction of the most important Phase I drug metabolizing enzymes, the cytochromes P450 (CYPs). To date, there have been limited reports regarding the inhibitory effects of Ashwagandha extracts on the CYPs, and no published studies examined whether Ashwagandha extracts have the potential to induce CYP expression. Ashwagandha is used by older adults in particular to ameliorate stress, insomnia, and cognitive deficiencies; and this population is often taking conventional medications for other health conditions. Thus, use by the elderly makes a study of how Ashwagandha affects CYP activity and expression an important concern. Ashwagandha is one of two botanicals being studied for its ability to improve resilience in the BENFRA Botanical Dietary Supplements Research Center (BDSRC) at Oregon Health and Science University (U19 AT010829). Our work will synergize with that of the BENFRA BDSRC, which will provide authenticated and chemically characterized extracts of Ashwagandha roots and leaves for testing in the applicant’s laboratory at the University of Auburn. For our project, we propose two Specific Aims. In Aim 1, we will screen different types of Ashwagandha extracts for inhibition of 9 human hepatic CYPs (including all CYPs recommended for drug-drug evaluation by the FDA). Inhibition of CYPs in pooled human liver microsomes will be measured using CYP probe substrates and our ultrafast UHPLC-MS/MS cocktail assay. In Aim 2, we will assess induction of CYP mRNA expression caused by Ashwagandha extracts using primary human hepatocytes in sandwich culture according to the FDA Guidance on in vitro drug-drug interactions studies. The resulting comprehensive data of Ashwagandha extract-CYP interactions will be made available through scientific publications. Demonstration of inhibition or induction of CYPs by Ashwagandha will be followed by future studies that seek to identify the compounds responsible for the observed effects. Demonstration of interactions with CYP enzymes will be followed by confirmatory clinical studies of Ashwagandha-drug pharmacokinetic interactions.