Human Cytochrome P450 4F Enzymes and Drug Interactions

人类细胞色素 P450 4F 酶和药物相互作用

• 批准号: 8304951
• 负责人: Zhuo Michael Wang
• 金额: $ 30.7万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-06 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8304951
• 关键词: Address; Adverse event; Anticoagulants; Anticoagulation; Arachidonic Acids; Biological Assay; Catabolism; Cells; Cholesterol; Clinical; Clinical Research; Cytochrome P450; Development; Dose; Double-Blind Method; Drug Interactions; Drug Kinetics; Enzymes; Exhibits; Foundations; Future; Genetic Crossing Over; Goals; Hemorrhage; Hepatocyte; High Pressure Liquid Chromatography; Human; Hydroxylation; In Vitro; Inflammatory; Laboratories; Leukotriene B4; Lovastatin; Mass Spectrum Analysis; Measures; Mediating; Metabolic; Metabolism; Methodology; Methods; Molecular; Nutrient; Pathway interactions; Pharmaceutical Preparations; Pharmacodynamics; Pharmacotherapy; Physiological; Placebo Control; Play; Prevalence; Principal Investigator; Property; Proteins; Proteomics; Public Health; Randomized; Recombinants; Reporting; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Testing; Time; Translational Research; Vitamin E; Vitamin K; Vitamin K 1; Warfarin; Work; base; carboxylation; clinically relevant; clinically significant; cofactor; dosage; gamma-glutamyl carboxylase; healthy volunteer; high risk; improved; innovation; novel; programs; protein expression; public health relevance; tandem mass spectrometry; therapeutic target

DESCRIPTION (provided by applicant): Drug-drug interactions play an important role in clinical adverse events due to the wide prevalence of multi-drug therapies. The human cytochrome P450 4F (CYP4F) subfamily of enzymes has recently been demonstrated to play a significant role in the metabolism of both endogenous compounds, including arachidonic acid and leukotrienes B4 (LTB4), and drugs/nutrients, including pafuramidine, fingolimod, and vitamin E. However, the role of CYP4F enzymes in mediating potential drug interactions remains uncharacterized. Numerous clinically significant adverse interactions, including major and fatal bleeding episodes, have been reported between the widely-prescribed cholesterol-lowering agents, the "statins", and the anticoagulant warfarin. Co-administration of these drugs is expected to be on the rise due in part to the purported pleiotropic effects of statins. An improved understanding of the molecular mechanisms underlying the statin-warfarin interaction will provide necessary scientific basis and methodologies to develop an interaction-free statin-warfarin combination, and allow enactment of pertinent regulatory measures limiting certain high-risk statin-warfarin combinations to be prescribed to hundreds of thousands of people each year, which has great public health importance. The long-term goal of the PI's research program is to further our understanding of the pharmacologic and physiologic roles of the human CYP4F enzymes. The specific goal of the proposed research is to develop a mechanistic understanding of the role of CYP4F2 in the interaction between statins and warfarin. An innovative and translational experimental approach will be employed to test the central hypothesis that certain statins induce CYP4F2-mediated vitamin K1 metabolism and potentiate the anticoagulant effect of warfarin in humans. Novel methodologies, including a mass spectrometry (MS)-based quantitative proteomic approach for protein quantification, a marker substrate activity assay for CYP4F2, and a double-blind, randomized, placebo-controlled, 2-period cross-over clinical study will be utilized to address key issues regarding 1) the deactivation of vitamin K1 upon ?-hydroxylation by CYP4F2; 2) the induction of CYP4F2 protein expression and enzymatic activity by statin treatment; and 3) the clinical relevance of the effect of statins on warfarin anticoagulation efficacy. Results obtained from the proposed translational research will serve as a foundation for understanding the pharmacologic and physiologic role of CYP4F enzymes, and will provide important information useful in the development of safer statins, and a mechanism by which approved statins can be rank-ordered according to their CYP4F2-inducing potential to guide warfarin dosage adjustment when administered concomitantly. The proposed work has the following specific aims: 1) identify and characterize the vitamin K1 metabolite (K1-?-OH) generated by CYP4F2; 2) determine the induction profiles of statins towards CYP4F2 in HepG2 cells and primary human hepatocytes; and 3) assess the influence of statin treatment on the anticoagulant effect of warfarin in healthy volunteers. PUBLIC HEALTH RELEVANCE: Drug-drug interactions play an important role in clinical adverse events due to the wide prevalence of multi-drug therapy. In spite of numerous reports of clinically significant adverse interactions, the co-medication of warfarin and statins has drastically expanded in the US over the last decades. This proposal seeks to demonstrate a novel molecular mechanism for the statin-warfarin interaction and provide a pathway to develop an interaction-free statin-warfarin combination.

描述（由申请人提供）：由于多药治疗的广泛流行，药物-药物相互作用在临床不良事件中起着重要作用。人类细胞色素P450 4F （CYP4F）亚家族酶最近被证明在内源性化合物（包括花生四烯酸和白三烯B4 (LTB4)）和药物/营养物质（包括帕夫拉脒、fingolimod和维生素e）的代谢中发挥重要作用。然而，CYP4F酶在介导潜在药物相互作用中的作用尚未确定。在广泛使用的降胆固醇药物“他汀类药物”和抗凝华法林之间，已经报道了许多临床显著的不良反应，包括严重和致命的出血事件。由于他汀类药物据称具有多效性，这些药物的联合用药预计将呈上升趋势。进一步了解他汀-华法林相互作用的分子机制将为开发无相互作用的他汀-华法林组合提供必要的科学基础和方法，并允许制定相关的监管措施，限制每年数十万人使用某些高风险的他汀-华法林组合，这对公共卫生具有重要意义。PI研究计划的长期目标是进一步了解人类CYP4F酶的药理学和生理学作用。该研究的具体目标是对CYP4F2在他汀类药物和华法林相互作用中的作用进行机制理解。我们将采用一种创新的转化实验方法来验证某些他汀类药物诱导cyp4f2介导的维生素K1代谢并增强华法林在人体中的抗凝作用的中心假设。新的方法，包括基于质谱（MS）的蛋白质定量蛋白质组学方法，CYP4F2的标记底物活性测定，以及双盲，随机，安慰剂对照，2期交叉临床研究，将用于解决以下关键问题：1)维生素K1的失活？CYP4F2 -羟基化；2)他汀类药物对CYP4F2蛋白表达和酶活性的诱导作用；3)他汀类药物对华法林抗凝疗效影响的临床意义。该转化研究的结果将为理解CYP4F酶的药理学和生理学作用奠定基础，并将为开发更安全的他汀类药物提供重要信息，以及根据其cyp4f2诱导潜能对已批准的他汀类药物进行排序的机制，以指导华法林同时给药时的剂量调整。提出的工作有以下具体目的：1)鉴定和表征维生素K1代谢物(K1-？-OH)由CYP4F2产生；2)确定他汀类药物在HepG2细胞和人原代肝细胞中对CYP4F2的诱导谱；3)评估他汀类药物治疗对健康志愿者华法林抗凝作用的影响。