Identification of CYP5122A1 inhibitors as chemical probes for Leishmania biology

鉴定 CYP5122A1 抑制剂作为利什曼原虫生物学化学探针

• 批准号: 10061546
• 负责人: Zhuo Michael Wang
• 金额: $ 47.82万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-12-21 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10061546
• 关键词: Affect; Afghanistan; Alleles; Anabolism; Azoles; Binding; Biochemical; Biological Assay; Biology; Bite; Case Study; Chemicals; Communicable Diseases; Complex; Country; Crystallization; Cutaneous Leishmaniasis; Cytochrome P450; Dangerousness; Disease; Drug Targeting; Enzyme Kinetics; Enzymes; Ergosterol; Excision; Exhibits; Fluorescence; Future; Gene Deletion; Gene Mutation; Growth; High Pressure Liquid Chromatography; Human; Hybrids; Hypersensitivity; Image; Impairment; Infection; Iraq; Ketoconazole; Knock-out; Laboratories; Lanosterol; Left; Leishmania; Leishmania donovani; Leishmaniasis; Life; Ligand Binding; Ligands; Malaria; Medical; Metabolic Biotransformation; Military Personnel; Organ; Organism; Outcome; Parasites; Parasitic Diseases; Pathogenicity; Pathway interactions; Pharmaceutical Preparations; Phlebotominae; Prodrugs; Proteins; Public Health; Reaction; Resistance; Roentgen Rays; Role; Route; Skin; Sterols; Structural Models; Structure; Testing; Vaccines; Visceral Leishmaniasis; assay development; base; chemotherapy; cost; cytotoxicity; drug discovery; effective therapy; high throughput screening; inhibitor/antagonist; innovation; mortality; new therapeutic target; novel; posaconazole; screening; side effect

Summary Human leishmaniasis is a devastating infectious disease caused by protozoan parasites belonging to the genus of Leishmania. The disease is found in more than 90 countries and responsible for an estimated 1-2 million new infections each year worldwide. Leishmaniasis is also common in returning U.S. military personnel from Iraq and Afghanistan, with more than two thousand cases reported since 2001. Leishmania parasites cause disfiguring skin sores (cutaneous leishmaniasis or CL) and life-threatening infection of vital internal organs (visceral leishmaniasis or VL). VL (~20% of all leishmaniasis cases) is the most dangerous and fatal form of the disease, with a mortality rate close to 100% if left untreated. It is the second most deadly parasitic disease in the world (after malaria). Despite being a serious public health problem in many endemic regions, there are no vaccines or preventative chemotherapies available for leishmaniasis control. Current antileishmanial drugs have limited efficacy, serious side effects and high cost. Hence, there is a major unmet medical need for safe and effective drugs against leishmaniasis. Leishmanial CYP5122A1 is a novel cytochrome P450 (CYP) enzyme that is essential for the survival of L. donovani, a major causative agent for VL. Independent and our own studies have led us to hypothesize that identification of selective CYP5122A1 inhibitors as chemical probes will allow elucidation of its important role in the ergosterol biosynthesis by Leishmania. To test our hypothesis, three specific aims are proposed to answer three main questions: 1) How do the biochemical roles of CYP5122A1 and CYP51 compare in the ergosterol biosynthesis pathway of Leishmania? 2) Can selective CYP5122A1 inhibitors be identified and validated? and 3) Are arylimidamides (AIAs) and/or AIA-azole hybrids potential chemical probes to help delineate biochemical roles of CYP5122A1 in Leishmania biology? Innovative experimental approaches will be employed in the proposed project, e.g., a new HPLC-MS/MS-based sterol assay, fluorescence-based CYP5122A1 and CYP51 inhibition assays, and cutting-edge high content imaging-based intracellular antileishmanial assays. If successful, our proposed project to identify novel CYP5122A1 inhibitors will elucidate this protein’s role in Leishmania biology and validate CYP5122A1 as a drug target. Such an outcome could have a major positive impact on long-term leishmaniasis control around the globe.

摘要 人类利什曼病是一种由原生动物寄生虫引起的破坏性传染病，属于 利什曼原虫属。这种疾病在90多个国家被发现，据估计有1-2 全球每年新增感染人数为100万人。利什曼病在回国的美国军人中也很常见 来自伊拉克和阿富汗，自2001年以来报告了2000多例病例。利什曼原虫 导致毁容皮肤溃疡(皮肤利什曼病或CL)和危及生命的重要内脏感染 器官(内脏利什曼病或VL)。VL(约占所有利什曼病病例的20%)是最危险和最致命的 这种疾病的形式，如果不治疗，死亡率接近100%。它是第二致命的寄生虫 世界上的疾病(在疟疾之后)。尽管在许多流行地区是一个严重的公共卫生问题， 目前还没有疫苗或预防性化学疗法可用于控制利什曼病。当前 抗利什曼病药物疗效有限，副作用严重，价格昂贵。因此，有一个重大的未满足 医疗需要安全有效的药物来治疗利什曼病。利什曼式细胞色素P5122A1是一种新的 细胞色素P450(CYP)酶，对杜氏乳杆菌的生存至关重要，它是 VL.独立的和我们自己的研究使我们假设选择性CYP5122A1的鉴定 作为化学探针的抑制剂将有助于阐明其在麦角甾醇生物合成中的重要作用 利什马尼亚。为了验证我们的假设，提出了三个具体目标来回答三个主要问题：1)如何 细胞色素P5122A1和细胞色素P51在麦角甾醇生物合成途径中的生化作用比较吗？ 利什曼病？2)选择性的CYP5122A1抑制剂能被识别和验证吗？和3)芳基咪胺类化合物 (AIAS)和/或AIA-唑杂交物有助于描述CYP5122A1的生化作用的潜在化学探针 利什马尼亚生物学吗？拟议的项目将采用创新的试验方法，例如 新的基于HPLC-MS/MS的甾醇检测，基于荧光的CyP5122A1和CyP51抑制检测，以及 尖端高含量成像细胞内抗利什曼试验。如果成功，我们的建议 发现新的CYP5122A1抑制剂的项目将阐明该蛋白在利什曼原虫生物学和 验证CYP5122A1作为药物靶点。这样的结果可能对长期经济产生重大积极影响 在全球范围内控制利什曼病。