In vitro screening of a unique botanical extract collection for herb-drug interaction (HDI) potential

体外筛选独特的植物提取物集合的草药-药物相互作用 (HDI) 潜力

• 批准号: 10226906
• 负责人: Bill Gurley
• 金额: $ 21.9万
• 依托单位: UNIVERSITY OF MISSISSIPPI
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10226906
• 关键词: ABCB1 gene; Affect; Architecture; Aromatic Hydrocarbons; Biological Availability; Botanical dietary supplements; Botanicals; CYP1A2 gene; CYP2C19 gene; CYP2C9 gene; CYP3A4 gene; Cell Line; Cellular Assay; Clinical; Collection; Commerce; Communities; Cytochrome P450; Data; Development; Diet; Drug Interactions; Drug Kinetics; Drug Prescriptions; Drug user; Enzymes; Equipment; Excretory function; Exhibits; Formulation; Fractionation; Future; Gene Expression; Gene Proteins; Goals; Goldenseal; Grant; Grapefruit juice; Health Professional; Hepatocyte; Herb-Drug Interactions; Human; Hypericum perforatum; In Vitro; Intestines; Investigation; Liver; Manufacturer Name; Mediating; Medical; Metabolic Biotransformation; Metabolism; Methodology; Methods; Mississippi; Molecular; Natural Product Drug; Natural Products; Nuclear Protein; Nuclear Receptors; Oral; Outcome; Pathway interactions; Pharmaceutical Preparations; Pharmacotherapy; Phase; Phytochemical; Piper nigrum; Plants; Population; Pregnanes; Primary carcinoma of the liver cells; Protein Isoforms; Public Health; Receptor Activation; Research; Risk; Safety; Sampling; Technical Expertise; Techniques; Technology; Testing; Translating; Universities; Xenobiotics; absorption; chemical fingerprinting; clinical risk; clinically relevant; constitutive androstane receptor; design; dietary supplements; drug efficacy; drug metabolism; exposed human population; hyperforin; in vitro activity; in vivo; new technology; novel; prevent; prospective; protein function; receptor; repository; screening; stem

PROJECT SUMMARY Herb-drug interactions (HDI) have been a significant public health concern for over a quarter century.1 Such concerns are bolstered by the fact that nearly 70% of the U.S. population takes some form of dietary supplement, and almost 30% of all prescription drug users take botanical dietary supplements (BDS) concomitantly. Currently tens of thousands of BDS are on the U.S. market, many containing phytochemicals rarely encountered in the normal diet. Because these unique phytochemicals utilize the same absorption, distribution, metabolism and excretion pathways as conventional medications, the chance for pharmacokinetic-mediated HDI (PK-HDI) is high. PK-HDI are those in which single or multiple phytochemicals modulate various human drug metabolizing enzymes and/or transporters (DMET) that, in turn, affect the biotransformation and disposition of concurrently administered medications.2 To date, only a few botanicals appear to pose risks for clinically relevant PK-HDI. Of these, St. John’s wort (SJW) is the most problematic as it renders many medications ineffective by inducing their metabolism and reducing their oral bioavailability. SJW’s tremendous HDI potential is due to hyperforin, a unique phytochemical that potently activates a key protein nuclear receptor (NR) known as the human pregnane xenobiotic receptor (hPXR). Like all NRs, hPXR upregulates the expression and activity of various DMET. Preliminary data from the University of Mississippi’s National Center for Natural Products Research (NCNPR) indicates that other botanicals also harbor phytochemicals capable of activating hPXR along with other important NRs like the aromatic hydrocarbon receptor (AhR) and constitutive androstane receptor (CAR). Thus, there is a critical need for systematic screening of popular BDS to determine their PK-HDI risk via NR activation. Equally important is the need for properly characterized botanical extracts in which to screen. Since 1995, the NCNPR has amassed one of the world’s largest botanical extract repositories. In this grant submission, we propose a 4- phased approach to screen 30 of the top-selling BDS for their ability to activate NRs (i.e., hPXR, AhR, CAR) using well-characterized ethanolic extracts from the NCNPR repository. (1) Initial high throughput NR screenings will utilize standard in vitro techniques employing transfected cell lines. (2) Extracts capable of activating NRs will be assessed for their ability to upregulate both gene expression and protein function for specific DMET (e.g., CYP1A2, CYP2C9, CYP2C19, CYP3A4, ABCB1) using standard human hepatoma and intestinal cell assays. (3) Culpable phytochemicals within candidate extracts will be isolated and identified. (4) Finally, in lieu of primary human hepatocyte cultures, extracts/phytochemicals exhibiting positive results from the first two screening phases will be evaluated for their PK-HDI risk using the novel Emulate™ human “Liver-on-Chip” technology. For extracts exhibiting positive results in all screening stages, a data “dossier” will be provided to the Center of Excellence for Natural Product Interaction Research for confirmatory analyses and possible future clinical PK- HDI studies.

项目总结 四分之一世纪以来，草药相互作用(HDI)一直是一个重大的公共卫生问题。 近70%的美国人服用某种形式的膳食补充剂，这一事实加剧了人们的担忧。 几乎30%的处方药使用者同时服用植物膳食补充剂(BDS)。目前 美国市场上有数以万计的BDS，其中许多含有在 正常饮食。因为这些独特的植物化学物质利用相同的吸收、分布、新陈代谢和 排泄途径与传统药物一样，药代动力学介导的HDI(PK-HDI)的机会是 很高。PK-HDI是指一种或多种植物化学物质对人体各种药物代谢的调节作用 酶和/或转运体(DMET)，进而影响同时存在的生物转化和处置 给药2到目前为止，似乎只有几种植物性药物对临床相关的PK-HDI构成风险。的 这些，圣约翰草(SJW)是最有问题的，因为它使许多药物无效，通过诱导他们的 并降低其口服生物利用度。SJW巨大的HDI潜力归功于金丝桃素，一种独特的 一种植物化学物质，能有效激活一种称为人类孕烷的关键蛋白质核受体(NR) 异种生物受体(HPXR)。与所有NRs一样，hPXR上调各种DMET的表达和活性。 密西西比大学国家天然产物研究中心(NCNPR)的初步数据 表明其他植物也含有能够激活hPXR的植物化学物质以及其他重要的 NRs类似于芳香烃受体(AhR)和构成雄烷受体(CAR)。因此，有一个 迫切需要对流行的BDS进行系统筛查，以确定其通过NR激活而发生PK-HDI的风险。同样 重要的是需要具有适当特征的植物提取物进行筛选。自1995年以来，NCNPR 已经积累了世界上最大的植物提取物储存库之一。在这份拨款申请中，我们建议4- 分阶段对最畅销的BDS中的30个进行筛选，以了解其激活NR的能力(即hPXR、AhR、CAR) 使用来自NCNPR储存库的特征良好的乙醇提取物。(1)初步高通量天然橡胶筛选 将利用标准的体外技术，使用转基因细胞系。(2)能够激活NRS的提取物 将评估它们上调特定DMET的基因表达和蛋白质功能的能力(例如， CYP1A2、CYP2C9、CYP2C19、CYP3A4、ABCB1)使用标准的人肝癌和肠道细胞检测。 (3)将分离和鉴定候选提取物中的有问题的植物化学物质。(4)最后，代替初选 前两次筛查显示阳性结果的人肝细胞培养、提取物/植物化学物质 将使用新的模拟™人类“芯片上的肝脏”技术来评估各阶段的PK-HDI风险。为 在所有筛选阶段显示阳性结果的摘录，将向美国疾病控制与预防中心提供数据档案 用于验证性分析和未来可能的临床PK的天然产物相互作用研究的卓越表现- HDI研究。