Validation of preclinical assays with clinical data to evaluate the safety of estrogenic botanical dietary supplements

使用临床数据验证临床前测定，以评估雌激素植物膳食补充剂的安全性

• 批准号: 10207475
• 负责人: Alan M Wong
• 金额: $ 1.86万
• 依托单位: OREGON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-09 至 2022-01-08
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10207475
• 关键词: Address; Adult; American; Anabolism; Animal Model; Biological; Biological Assay; Biological Availability; Biological Markers; Botanical dietary supplements; Botanicals; Cell Line; Cell model; Cells; Cellular Assay; Chemicals; Cimicifuga racemosa; Clinical; Clinical Data; Clinical Research; Clinical Trials; Complement; Complex; Consumption; Data; Detection; Disease; Drops; Estrogen Metabolism; Estrogens; Goals; Health; Hepatocyte; Hormones; Hot flushes; Human; Humulus; Impaired cognition; In Vitro; Laboratories; Laboratory Research; Liquid Chromatography; Malignant Neoplasms; Manufacturer Name; Marketing; Medicine; Menopausal Symptom; Menopause; Metabolism; Methodology; Methods; Modeling; Natural Products; Non-Prescription Drugs; Outcome; Perimenopause; Phase I Clinical Trials; Phase II Clinical Trials; Phytoestrogens; Postmenopause; Preparation; Proteomics; Publishing; Regulation; Reporting; Research; Research Personnel; Risk; Safety; Sales; Sampling; Serum; Specimen; Steroid biosynthesis; Time; Translating; Trifolium pratense; United States; United States Food and Drug Administration; United States National Institutes of Health; Urine; Validation; Weight Gain; Woman; Women' s Health; Xenobiotics; age group; analytical method; cardiovascular disorder risk; clinical center; cognitive function; cost; design; dietary supplements; estrogenic; hormone therapy; improved; in vitro Assay; in vivo; innovation; instrumentation; metabolomics; pre-clinical; predictive modeling; predictive test; reagent standard; safety study; standard of care; symptom management; tandem mass spectrometry

ABSTRACT More than half of the top selling botanicals dietary supplements (BDS) are marketed to peri- and post- menopausal women. Approximately 70% of women in this age group reported having consumed BDS to manage symptoms or treat disease. The focus of this project is to address the safety of these popular BDS. We hypothesize that most BDS used by women to manage menopausal symptoms are safe, but some botanicals might pose risk through modulation of estrogen biosynthesis and metabolism. By carrying out post-analyses of urine and serum from our completed Phase I and Phase II clinical trials of black cohosh (Actaea racemosa), hops (Humulus lupulus) and red clover (Trifolium pratense), we will confirm possible modulation to hormone levels. Two in vitro cell assays will be validated to model estrogen biosynthesis and estrogen metabolism. Our three Specific Aims will include the following: 1) Develop fast liquid chromatography-tandem mass spectrometry analytical methods with improved sensitivity and selectivity for the quantitation of estrogens and their metabolites. The goal is to minimize sample preparation and analysis time and utilize only commercially available derivatization reagents and standard instrumentation. The methods will be validated for the analysis of serum, urine, and cell media samples. 2) Establish in vitro assays for identifying botanical modulations to estrogen biosynthesis and estrogen metabolism. Two cell lines widely used in research laboratories for regulatory purposes will be evaluated for their biological relevance and availability. 3) Quantify estrogens and their metabolites in clinical samples of women taking BDS. This will enable us to determine significant modulations to hormone levels and be used to validate the in vitro assays from Aim 2. Together, the clinical data will answer our hypothesis regarding the importance of estrogen modulation in BDS product safety and mechanism of action, and the validated in vitro assays will provide a fast and accurate preclinical mechanism to assess BDS safety at modest cost.

摘要 超过一半的最畅销的植物性膳食补充剂（BDS）是销售给老年人和老年人的。 更年期妇女在这个年龄组中，大约70%的妇女报告说， 症状或治疗疾病。该项目的重点是解决这些流行的BDS的安全性。我们 假设大多数女性用于管理更年期症状的BDS是安全的，但一些植物性药物 可能通过调节雌激素的生物合成和代谢造成风险。通过对以下方面进行事后分析， 来自我们完成的黑升麻（Actaea racemosa）的I期和II期临床试验的尿液和血清， 啤酒花（Humulus lupulus）和红三叶草（Trifolium pratense），我们将确认可能的调节激素 程度.将验证两种体外细胞试验以模拟雌激素生物合成和雌激素代谢。我们 具体目标包括：1）发展快速液相色谱-串联质谱 具有改进的灵敏度和选择性的用于定量雌激素及其 代谢物。目标是最大限度地减少样品制备和分析时间， 衍生试剂和标准仪器。将对用于血清分析的方法进行验证， 尿液和细胞培养基样本。2)建立鉴定植物对雌激素调节作用的体外试验 生物合成和雌激素代谢。两种细胞系广泛用于研究实验室的调节 将评估其生物相关性和可用性。3)量化雌激素及其 服用BDS的女性临床样本中的代谢物。这将使我们能够确定重要的调制， 激素水平，并用于验证Aim 2的体外试验。临床数据将共同回答 我们关于雌激素调节在BDS产品安全性和作用机制中的重要性的假设， 经验证的体外试验将提供一种快速准确的临床前机制，以评估 价格适中。