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

描述（由申请人提供）：细胞色素P450 2E1 （CYP2E1）是一种含血红素的单加氧酶，可催化许多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。CYP2E1在人肝脏中被酒精高度诱导。酒精对CYP2E1的上调增强了肝毒性，增加了酗酒者患癌症的风险。研究认为，CYP2E1介导的氧化应激和脂质过氧化在酒精性肝损伤的发病机制中起重要作用。抑制CYP2E1活性已被证明可以将酒精的毒性降至最低。然而，现有的CYP2E1抑制剂由于其混杂性和毒性而不能在体内使用。有必要开发特异性和无毒的CYP2E1抑制剂，既可用于基础研究，以调查CYP2E1在酒精相关疾病中的具体作用，又可用于临床治疗和预防酒精中毒。我假设，通过虚拟高通量筛选（vHTS），可以从小配体文库中鉴定出有效的、特异性的CYP2E1机制抑制剂。在本提案中，我计划以两个具体目标来验证这一假设：1)使用双重选择标准在小配体文库中筛选大约55,000个乙炔化合物；2)通过生化分析vHTS命中的效力和特异性。该提案的结果将产生大约12种CYP2E1先导抑制剂，可用于加速治疗剂的开发。

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.