Regulation of hepatic uptake transporters by tyrosine kinases

酪氨酸激酶对肝摄取转运蛋白的调节

• 批准号: 10366185
• 负责人: Jason A Sprowl
• 金额: $ 47.06万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 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/10366185
• 关键词: Adverse event; Affect; American; Angiotensin-Converting Enzyme Inhibitors; Binding; Biological; 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; Structure; 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/antagonist; 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

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 但是 介导这些转运蛋白功能的翻译后修饰的表征仍然存在 大大研究了。因此，与该提议相关的长期目标是定义范围和 翻译后介导的药理学上重要药物转运蛋白调节的重要性，例如 OATP1B1。该提议的总体目的是描述酪氨酸激酶抑制剂的后果 - （TKI）介导OATP1B1的抑制作用。中心假设是特定的酪氨酸激酶是必不可少的 OATPB1的调节剂，并且对此过程的干扰将减少肝积累并增加 OATP1B1底物的血浆浓度，导致药代动力学（PK）和药效学（PD） 药物相互作用（DDIS）。该项目的理由是阐明监管机制 影响OATP1B1功能将有助于改进迄今为止的治疗策略 产生了各种意外的DDI介导的威胁生命的毒性，其中包括他汀类药物诱导的 横纹肌溶解。中心假设将通过追求两个具体目标来检验：1）识别和阐明 涉及OATP1B1功能的调节激酶的作用； 2）研究调节的体内效应 OATP1B1活性的激酶。在第一个目标下，将使用基于LC/MS/MS/MS的蛋白质组学来测量 激酶抑制剂能够在体外和体内调节OATP1B1磷酸化，而遗传操纵 将利用全面的激酶抑制或结合测定法确定调节激酶 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 - 缺乏体内小鼠模型将用于评估OATP1B1底物的PK/PD变化 或缺乏TKI。这将提供有关酪氨酸激酶在调节OATP1B1--的作用的机械洞察力。 介导的肝脏处理底物。 AIM 2的集体发现也将用于开发 基于生理的PK/PD模型，以预测TKI AOTP1B1底物相互作用的结果。研究 在本申请中提出的是创新的，因为它代表了与现状的实质性不同 探索酪氨酸激酶活性作为OATP1B1功能的调节剂，并作为全身水平的介体 内源性生物标志物和异种生物。拟议的研究很重要，因为它有望 对OATP1B1（分子）功能必不可少的调节机制提供重要的新见解 激酶途径调节该转运蛋白及其在许多相关的肝脏处理中的作用 内源性和异种生物底物。最终，这种知识有望为基金会做出贡献 防止患者威胁生命的DDI。