Virtual High Throughput Screening: Specific Mechanism-based Inhibitors of CYP2E1

说明(申请人提供)：细胞色素P450 2E1(细胞色素P450 2E1)是一种含血红素的单加氧酶，可催化多种肝毒素和致癌物的氧化，包括乙醇、对乙酰氨基酚、亚硝胺和四氯化碳等。酒精对人体肝脏有很强的诱导性。酒精上调了细胞色素P4502的表达，增加了酒精对肝脏的毒性，增加了酒精者患癌症的风险。目前认为，在酒精性肝损伤的发病机制中，细胞色素P450 2 E1I在其介导的氧化应激和脂质过氧化中起重要作用。研究表明，抑制细胞色素P450酶2的活性可以将酒精的毒性降到最低。然而，现有的CYP2E1抑制剂由于混杂和毒性，不能在体内使用。有必要开发特异性的、无毒的CYP2E1抑制剂，既可用于基础研究，以研究CYP2E1在酒精相关疾病中的特定作用，也可用于临床治疗和预防酒精中毒。我推测，通过虚拟高通量筛选(VHTS)，可以从小的配基文库中鉴定出有效的、基于特定机制的CYP2E1抑制剂。在这个提议中，我计划用两个具体的目标来检验这个假设：1)使用双重选择标准在小的配体库中筛选大约55,000个乙炔类化合物；2)通过生化分析来分析vHTS点击的效力和特异性。这项提案的结果将产生大约12种可用于加速治疗药物开发的CYP2E1的先导抑制剂。 与公共健康相关：长期饮酒会导致一系列健康问题，包括大脑损伤、胎儿酒精综合征、癌症风险增加和酒精性肝病。因此，预防和治疗与酒精有关的疾病对公众健康至关重要。在这个项目中，我建议找出新的、特异性的细胞色素P450-2的抑制物。长期目标是开发可用于临床治疗和预防酒精性肝病的CYP2E1特异性抑制剂。该方法也可用于识别其他细胞色素P450靶标的特异性抑制剂。

DESCRIPTION (provided by applicant): Cytochrome P450 2E1 (CYP2E1) is a heme-containing monooxygenase that catalyzes the oxidation of a number of hepatotoxins and procarcinogens including ethanol, acetaminophen, nitrosamines, and carbon tetrachloride, among many others. CYP2E1 is highly inducible in human livers by alcohol. Upregulation of CYP2E1 by alcohol enhances hepatotoxicity and increases the risk of developing cancer in alcoholics. It is thought that CYP2E1 plays an important role in the pathogenesis of alcohol-induced liver injury due to CYP2E1-mediated oxidative stress and lipid peroxidation. Inhibition of CYP2E1 activity has been shown to minimize the toxicity of alcohol. However, existing CYP2E1 inhibitors cannot be used in vivo because of their promiscuity and toxicity. There is a need to develop specific and non-toxic CYP2E1 inhibitors that can be used both for basic research to investigate the specific role of CYP2E1 in alcohol-related diseases and for use in the clinic to treat and prevent alcoholism. I hypothesize that potent and specific mechanism-based inhibitors of CYP2E1 can be identified from small ligand libraries through virtual high throughput screening (vHTS). In this proposal, I plan to test this hypothesis with two specific aims: 1) to screen approximately 55,000 acetylenic compounds in small ligand libraries using dual selection criteria and 2) to analyze the potency and specificity of the vHTS hits by biochemical assays. The outcome of this proposal will yield approximately a dozen lead inhibitors for CYP2E1 that can be used to accelerate the development of therapeutic agents. PUBLIC HEALTH RELEVANCE: Chronic alcohol consumption leads to a host of health issues including brain impairment, fetal alcohol syndrome, increased risk of cancer, and alcohol liver diseases. Therefore, the prevention and treatment of alcohol-related diseases are critically important to public health. In this project, I propose to identify new, specific inhibitors for CYP2E1. The long-term goal is to develop CYP2E1-specific inhibitors that can be used clinically to treat and prevent alcohol liver diseases. This approach may also be applied to identify specific inhibitors for other cytochrome P450 targets.