RESVERATROL BIOAVAILABILITY--PRECLINICAL AND CLINICAL

描述：(申请人描述)流行病学研究，以及 对实验动物的研究表明，水果中存在的多酚， 蔬菜和饮料对多种癌症都有保护作用。一个 迅速增长的生化研究表明，大量可信的 产生这些效应的机制。然而，目前还不清楚在多大程度上以及在何种程度上 形成膳食多酚的物质在人体肠道中被吸收 因此可以获得对建议的蜂窝动作点的访问。作为临床和临床应用 对多酚类物质生物利用度的动物研究一般都是 不成功，我们正在使用一种中间的临床前方法，使用 Caco-2细胞作为人体肠道吸收模型的实验研究 新陈代谢。拟议的研究重点是多酚白藜芦醇，一种 葡萄和红酒中的植物抗毒素。我们的工作假设是Caco-2 细胞模型可以很好地预测药物的吸收和代谢 人体内的白藜芦醇。 在目标1中，我们将第一手确定吸收，即根尖到 白藜芦醇在Caco-2细胞单层的基侧转运 一种分子特异性的高效液相色谱分析方法。这也将产生代谢的命运 白藜芦醇，包括潜在的氧化和结合产物， 用高效液相色谱/质谱仪等光谱方法进行鉴定。体内的剂量依赖关系 运输和新陈代谢将特别令人担忧。 解吸附的相应研究，即根尖运输的基侧， 以确定可能的定向运输 白藜芦醇及其代谢产物由外排泵介导，如 P-糖蛋白和MRPs。此外，我们还将对药物进行鉴定 参与白藜芦醇生物转化的代谢酶。在《目标2》中，我们 将直接决定白藜芦醇的口服生物利用度。这 需要口服(25毫克)和静脉注射(1 Mg)剂量。将给予50microCi的(14C)标记的白藜芦醇 以使确定恢复和完全恢复成为可能 白藜芦醇的代谢命运。这也将使评估成为可能 排泄物，是多酚类物质清除的主要途径。从这些 我们将确定药代动力学参数的数据 白藜芦醇，包括清除(总、肾、代谢、粪便)， 消除分布的半衰期和体积。 AIMS 1和AIMS 2一起将让我们确定Caco-2细胞的用途 作为临床前模型。

DESCRIPTION: (Applicant's Description) Epidemiological studies, as well as studies in laboratory animals, suggest that polyphenols, present in fruits, vegetables and beverages, have protective effects in a variety of cancers. A rapidly growing number of biochemical studies show a vast array of plausible mechanisms for these effects. However, it is not clear to what extent and in which form the dietary polyphenols are absorbed in the human intestine and thus can gain access to proposed cellular sites of action. As clinical and animal studies of the bioavailability of the polyphenols in general have been unsuccessful, we are using an intermediary preclinical approach, using the Caco-2 cell line as model of human intestinal absorption as well as metabolism. The proposed study is focusing on the polyphenol resveratrol, a phytoalexin in grapes and red wine. Our working hypothesis is that the Caco-2 cell model is highly predictive of both absorption and metabolism of resveratrol in humans in vivo. In Aim 1 we will in first hand determine the absorption, i.e. apical to basolateral transport across the Caco-2 cell monolayer, of resveratrol, using a molecularly specific HPLC assay. This will also yield the metabolic fate of resveratrol, including potential oxidation as well as conjugation products, identified by HPLC/MS and other spectroscopic methods. Dose-dependency in transport and metabolism will be of particular concern. Corresponding studies of ex sorption, i.e. basolateral to apical transport, will be done in order to determine the possible directional transport of resveratrol and its metabolites mediated by efflux pumps, such as P-glycoprotein and the MRPs. In addition, we will identify the drug metabolizing enzymes involved in resveratrol biotransformation. In Aim 2 we will in first hand determine the oral bioavailability of resveratrol. This will require the administration of both an oral (25 mg) and an intravenous (1 mg) dose. Fifty microCi of (14C)-labeled resveratrol will be administered with each dose to make it possible to determine recoveries and the complete metabolic fate of resveratrol. This will also make it possible to assess fecal elimination, the major route of elimination of polyphenols. From these data we will determine the pharmacokinetic parameters for resveratrol, including clearances (total, renal, metabolic, fecal), elimination half -life and volume of distribution. Together Aims 1 and 2 will let us determine the usefulness of the Caco-2 cells as a preclinical model.