Effects of digestion on composition and bioavailability of compounds from Ashwagandha: an in vitro study

摘要 Withania somnifera，长期以来在阿育吠陀医学中以其药用特性而闻名。近年来， 消费W。在西方文化中，催眠药产品的销售量激增， 作为膳食补充剂通过零售店销售，通用名称为Ashwagandha。NCCIH赞助 BENFRA植物膳食补充剂研究中心（U19 AT 010829）目前正在研究 Ashwagandha，以确定其对年龄相关的神经系统恶化的影响。与制药药物不同， 在公开发行之前有一个广泛的评估过程，很少有人知道如何 Ashwagandha提取物中存在的化学物质的复杂混合物一旦被消耗并通过， 通过消化环境。该建议是创新的，因为它将评估代谢的稳定性， Ashwagandha提取物首次使用体外实验室模型模拟胃肠道 环境该项目还将评估哪些原始或修改后的组件能够通过肠道 通过被动扩散，大多数化合物从肠道进入循环的机制。 这项研究的中心假设是，Ashwagandha衍生代谢物的组成进入 循环与原始提取物的循环明显不同。这一假设将通过 具体目标：1。确定胃肠道环境对代谢物谱的影响 通过将提取物暴露于模拟胃肠液和酶来提取Ashwagandha;和2.识别 来自Ashwagandha原始提取物的代谢物的渗透性和使用合成的 肠膜模型。我们将使用这些体外模型系统的组合， 对照品和足够的重复样本进行稳健的统计分析，以标准化这些初步调查。超- 结合高分辨率质谱的压力液相色谱法将用于分析提取物， 孵育物和扩散室培养基，以评价化学结构和相对丰度的变化 Ashwagandha代谢物在这些复杂的矩阵的第一次。这些研究的结果将提供 深入了解一个更现实的Ashwagandha代谢谱后胃肠道消化，然后可以 使用已建立的临床前测定进一步合作研究。本研究的结果也可能 突出了目前尚未研究生物活性感兴趣的可渗透代谢物。从长远来看，这 该项目将建立一个强大的分析策略，可广泛用于评估其他药物的口服生物利用度 植物补充剂在体外环境中，在时间，劳动和资源密集型体内研究之前。

Abstract Withania somnifera, has long been known in Ayurvedic medicine for its medicinal properties. In recent years, consumption of W. somnifera products has skyrocketed in western cultures where they are now marketed through retail outlets as dietary supplements, under the common name, Ashwagandha. The NCCIH-sponsored BENFRA Botanical Dietary Supplements Research Center (U19 AT010829) is currently researching Ashwagandha, to determine its effects on age-related neurological deterioration. Unlike pharmaceutical drugs which have an extensive evaluation process prior to public distribution, very little is known regarding how the complex mixture of chemicals present in Ashwagandha extracts behaves once it is consumed and passes through the digestive environment. This proposal is innovative because it will evaluate metabolic stability of Ashwagandha extracts for the first time using in vitro laboratory models simulating the gastrointestinal environment. The project will also assess which original or modified components are able to pass the intestinal membrane via passive diffusion, the mechanism by which most compounds enter the circulation from the gut. The central hypothesis of this research is that the composition of Ashwagandha derived metabolites entering circulation is markedly different from that of the original extract. This hypothesis will be tested through the following specific aims: 1. Determine effects of gastrointestinal environments on the metabolite profiles of Ashwagandha extracts by exposing extracts to simulated gastro-intestinal fluids and enzymes; and 2. Identify permeabilities of metabolites from Ashwagandha original extracts and digestive incubations using a synthetic model of the intestinal membrane. We will use a combination of these in vitro model systems, appropriate controls, and sufficient replicates for robust statistical analyses to standardize these initial investigations. Ultra- pressure liquid chromatography coupled with high resolution mass spectrometry will be used to analyze extracts, incubates, and diffusion chamber media, to evaluate changes in chemical structures and relative abundances of Ashwagandha metabolites in these complex matrices for the first time. Results from these studies will provide insight into a more realistic Ashwagandha metabolic profile post-gastrointestinal digestion that can then be further studied collaboratively using established pre-clinical assays. Results of the present study may also highlight permeable metabolites of interest not currently under investigation for bioactivity. In the long term, this project will establish a robust analytical strategy that can be used broadly to assess oral bioavailability of other botanical supplements in an in vitro setting, prior to time, labor, and resource intensive in vivo studies.