Regulation of hepatic uptake transporters by tyrosine kinases

酪氨酸激酶对肝摄取转运蛋白的调节

基本信息

批准号：
10482372
负责人：
Jason A Sprowl
金额：
$ 45.99万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-15 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10482372
关键词：
Adverse event Affect American Angiotensin-Converting Enzyme Inhibitors Binding Biological Assay Biological Markers Biological Process Cell model Clinical Dangerousness Drug Interactions Drug Kinetics Drug Prescriptions Drug usage Enzymes Exposure to Foundations Goals Hepatic Hepatocyte Human In Vitro Investigational Drugs Knowledge LYN gene Lead Life Liver Mass Spectrum Analysis Measures Mediating Mediator of activation protein Metabolism Methotrexate Mission Modeling Molecular Mus Mutation National Institute of General Medical Sciences OATP Transporters Organic Cation Transporter POU2F2 gene Pathway interactions Patient-Focused Outcomes Patients Pharmaceutical Preparations Pharmacodynamics Pharmacologic Substance Pharmacology Phosphorylation Phosphorylation Site Phosphotransferases Physiological Plasma Positioning Attribute Post-Translational Protein Processing Post-Translational Regulation Process Protein Tyrosine Kinase Proteins Proteomics Public Health RNA interference screen Regulation Reporting Research Rhabdomyolysis Role Site Substrate Interaction Testing Therapeutic Toxic effect Transgenic Organisms Tyrosine Tyrosine Kinase Inhibitor Tyrosine Phosphorylation United States Food and Drug Administration Xenobiotics absorption base design experience genetic manipulation genetic variant in vivo inhibitor innovation insight kinase inhibitor knock-down liquid chromatography mass spectrometry mouse model novel therapeutics outcome prediction overexpression patient response pharmacodynamic model pharmacokinetics and pharmacodynamics prevent rosuvastatin src-Family Kinases therapeutic development treatment response treatment strategy uptake

项目摘要

ABSTRACT Hepatic drug uptake transporters are major regulators of systemic clearance and patient outcome, however, characterization of post-translational modifications that mediate the function of these transporters have remained greatly under studied. Therefore, the long-term goal associated with this proposal is to define the extent and significance of post-translational-mediated regulation of pharmacologically important drug-transporters, such as OATP1B1. The overall objective of this proposal is to delineate the consequences of tyrosine kinase inhibitor- (TKI) mediated inhibition of OATP1B1. The central hypothesis is that specific tyrosine kinase(s) are essential regulators of OATPB1, and that interference with this process will decrease hepatic accumulation and increase plasma concentrations of OATP1B1 substrates, resulting in pharmacokinetic (PK) and pharmacodynamic (PD) drug-drug interactions (DDIs). The rationale for this project is that elucidation of the regulatory mechanisms that affect OATP1B1 function will contribute to refinement of therapeutic strategies that have unfortunately to date yielded a variety of unexpected DDI-mediated life-threatening toxicities, which includes statin-induced rhabdomyolysis. The central hypothesis will be tested by pursuing two specific aims: 1) Identify and elucidate the role of regulatory kinases involved in OATP1B1 function; and 2) Investigate the in vivo effects of regulatory kinases on OATP1B1 activity. Under the first aim, LC/MS/MS-based proteomics will be used to measure the ability of kinase inhibitors to modulate OATP1B1 phosphorylation in vitro and ex vivo, while genetic manipulation and comprehensive kinase inhibition or binding assays will be utilized to identify the regulatory kinase of OATP1B1 function. Under the second aim, human transgenic OATP1B1, as well as transporter- and kinase- deficient in vivo mouse models will be used to assess PK/PD changes of OATP1B1 substrates in the presence or absence of TKIs. This will provide mechanistic insight into the role of tyrosine kinases in regulating OATP1B1- mediated hepatic handling of substrates. The collective findings of Aim 2 will also be utilized to develop physiologically-based PK/PD models to predict outcome of TKI-OATP1B1 substrate interactions. The research proposed in this application is innovative because it represents a substantive departure from the status quo by exploring tyrosine kinase activity as a regulator of OATP1B1 function and as a mediator of systemic levels of both endogenous biomarkers and xenobiotics. The proposed research is significant because it is expected to provide important new insights into regulatory mechanisms essential to the function of OATP1B1, the molecular kinase pathways modulating this transporter, and their role in the hepatic handling of the many relevant endogenous and xenobiotic substrates. Ultimately, such knowledge is expected to contribute to the foundation of preventing life-threatening DDIs in patients.

抽象的肝脏药物摄取转运蛋白是全身清除和患者结果的主要调节者，然而，介导这些转运蛋白功能的翻译后修饰的特征仍然存在很大程度上正在被研究。因此，与该提案相关的长期目标是确定范围和翻译后介导的药理学重要药物转运蛋白调节的重要性，例如 OATP1B1。该提案的总体目标是描述酪氨酸激酶抑制剂的后果- (TKI) 介导的 OATP1B1 抑制。中心假设是特定的酪氨酸激酶是必需的 OATPB1 的调节因子，干扰这一过程会减少肝脏蓄积并增加 OATP1B1 底物的血浆浓度，导致药代动力学 (PK) 和药效学 (PD) 药物间相互作用（DDI）。该项目的基本原理是阐明监管机制影响 OATP1B1 功能将有助于完善迄今为止不幸的治疗策略产生了多种意想不到的 DDI 介导的危及生命的毒性，其中包括他汀类药物引起的横纹肌溶解症。将通过追求两个具体目标来检验中心假设：1）识别并阐明参与 OATP1B1 功能的调节激酶的作用； 2) 研究监管的体内影响 OATP1B1 活性的激酶。在第一个目标下，基于 LC/MS/MS 的蛋白质组学将用于测量激酶抑制剂在体外和离体调节 OATP1B1 磷酸化的能力，同时进行基因操作和全面的激酶抑制或结合测定将用于鉴定调节激酶 OATP1B1 功能。在第二个目标下，人类转基因 OATP1B1 以及转运蛋白和激酶缺陷体内小鼠模型将用于评估 OATP1B1 底物存在时的 PK/PD 变化或缺乏 TKI。这将为酪氨酸激酶在调节 OATP1B1- 中的作用提供机制见解。介导的肝脏对底物的处理。目标 2 的集体发现也将用于开发基于生理学的 PK/PD 模型来预测 TKI-OATP1B1 底物相互作用的结果。研究本申请中提出的方案具有创新性，因为它代表了对现状的实质性背离探索酪氨酸激酶活性作为 OATP1B1 功能的调节剂和全身水平的调节剂内源性生物标志物和外源性生物标志物。拟议的研究意义重大，因为预计为 OATP1B1 功能所必需的调节机制提供重要的新见解，OATP1B1 是分子调节该转运蛋白的激酶途径及其在肝脏处理许多相关物质中的作用内源性和外源性底物。最终，这些知识有望为基金会做出贡献预防危及患者生命的 DDI。