Validation of Trypanosoma cruzi dihydroorotate dehydrogenase as a drug target for Chagas´disease.

验证克氏锥虫二氢乳清酸脱氢酶作为恰加斯病的药物靶点。

• 批准号: 10216399
• 负责人: Flavio da Silva Emery
• 金额: $ 13.49万
• 依托单位: UNIVERSITY OF SAO PAULO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-21 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10216399
• 关键词: Acrylamides; Active Sites; Acute; Address; Affect; Anabolism; Animal Disease Models; Antiparasitic Agents; Binding; Biological Assay; Blood; Brazil; Cell Culture Techniques; Cells; Chagas Disease; Chemical Structure; Chemicals; Chronic; Chronic Phase; Collaborations; Communities; Complex; Computer Assisted; Computer Models; Country; Crystallization; Cysteine; DHODH gene; Data; Developing Countries; Development; Dihydroorotate dehydrogenase; Disease; Dose; Drug Kinetics; Drug Targeting; Enzymatic Biochemistry; Enzyme Inhibition; Enzymes; Equilibrium; Etiology; Flavin Mononucleotide; Fumarates; Funding; Generations; Genetic study; Goals; Hot Spot; Infection; Institution; Kinetics; Knowledge; Latin America; Lead; Libraries; Ligands; Mammalian Cell; Maps; Measures; Metabolic Activation; Metabolism; Methods; Nitriles; Outcome; Oxidation-Reduction; Parasitic Diseases; Parasitology; Pathogenesis; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Play; Poverty; Predisposition; Property; Public Health; Pyrimidine; Pyrimidine Nucleotides; Reaction; Regimen; Research; Research Personnel; Role; Route; Running; Societies; Solvents; Structure; Succinate Dehydrogenase; Succinates; Techniques; Testing; Time; Toxic effect; Treatment Protocols; Trypanosoma cruzi; United States; Validation; X-Ray Crystallography; base; computational chemistry; cytotoxicity; data portal; design; dihydroorotate; divinyl sulfone; drug development; drug discovery; drug metabolism; effective therapy; efficacy evaluation; fighting; flexibility; in silico; in vitro activity; in vivo; inhibitor/antagonist; lead optimization; mouse model; multidisciplinary; neglect; new therapeutic target; novel; orotate; oxidation; public-private partnership; resistant strain; screening; side effect; structural biology; tool

ABTRACT Trypanosoma cruzi is the etiological agent of Chagas disease, a pathogenesis that affects 6 to 7 million people worldwide, mainly in Latin America. There is no effective treatment for chronic Chagas disease and resistant strains to the current frontline treatment have already emerged. There is a growing need for new pharmacological alternatives to treat this disease. This proposal exploits a target-based approach to search for lead compounds against Chagas disease. Trypanosoma cruzi dihydroorotate dehydrogenase (TcDHODH) is a flavin mononucleotide containing enzyme, which catalyzes the oxidation of L-dihydroorotate to orotate, the fourth step and only redox reaction in the de novo biosynthesis of pyrimidine nucleotides. TcDHODH was also described as a soluble fumarate reductase playing a role in connecting succinate/fumarate metabolism to de novo pyrimidine biosynthesis. DHODH has already been extensively exploited as a drug target for proliferative and parasitic diseases. Genetic studies have shown that DHODH is essential for T. cruzi survival providing evidence that this enzyme is an attractive target for the development of antichagasic drugs. The goal of this project is to chemically validate TcDHODH as a new drug target for Chagas disease and to provide leads for drug development. Our approach combines multiple techniques including computational chemistry, enzymology, structural biology, parasitology, and medicinal chemistry to develop selective and covalent inhibitors of TcDHODH. Initially, we will seek to gain a greater understanding of the DHODH hot spots by using solvent mapping to characterize the binding pockets with the flexible active-site loop open and closed. This information will help strengthen the computational modeling, and subsequent compound design. Covalent inhibition of enzymes based on the identification of a set of cysteine targeting covalent warheads including acrylamides, vinyl sulfones and nitriles, has been shown to be a successful approach in drug discovery pipelines. In our proposal, an initial set of putative inhibitors that incorporate an acrylamide warhead have already been rationally designed to target the TcDHODH active-site cysteine. Predicted inhibitors will be evaluated for inhibitory potency and mechanism of inhibition against TcDHODH, cytotoxicity and its anti-parasitic effects in cell culture. X-ray crystallography, fragment screening and medicinal chemistry will be combined to provide the chemical basis for the synthesis of a new generation of potent, selective, and drug-like inhibitors. The top 3 inhibitors that meet lead criteria (including PK and tolerability) will have their efficacy evaluated by DNDi via an in-kind contribution to the project. This proposal represents an unprecedent initiative to significantly contribute to building capacity in the field of drug discovery in Brazil, a field of research still very incipient in developing countries.

添加 克鲁齐锥虫是查加斯病的病因学药，这种发病机理影响了6至700万 全球人，主要在拉丁美洲。没有有效的慢性查加斯病和 对当前一线治疗的抗性菌株已经出现。对新需求越来越多 治疗这种疾病的药理替代方法。 该提案利用了一种基于目标的方法来寻找针对Chagas的铅化合物 疾病。锥虫二氢苯二酸锥虫脱氢酶（TCDHODH）是黄素单核苷酸 含有酶，酶催化l-二氢酯的氧化为orotate，第四步，仅 嘧啶核苷酸的从头生物合成中的氧化还原反应。 tcdhodh也被描述为 可溶性富马酸还原酶在将琥珀酸/富马酸盐代谢连接到从嘧啶的连接中发挥作用 生物合成。 Dhodh已经被广泛利用为增生和寄生虫的药物靶标 疾病。遗传学研究表明，Dhodh对于克鲁兹的生存至关重要，提供证据表明 该酶是抗挑剔药物开发的有吸引力的靶标。 该项目的目的是化学验证TCDHODH作为Chagas病的新药物 并为药物开发提供潜在客户。我们的方法结合了多种技术，包括 计算化学，酶学，结构生物学，寄生虫学和药物化学的发展 TCDHODH的选择性和共价抑制剂。最初，我们将寻求对 Dhodh热点通过使用溶剂映射来表征具有柔性活性位点的结合口袋 循环打开和关闭。这些信息将有助于加强计算建模，然后进行 复合设计。 基于鉴定一组半胱氨酸靶向共价的酶的共价抑制 弹头在内 药物发现管道。在我们的提案中，一组掺入丙烯酰胺的推定抑制剂 弹头已经经过合理设计，旨在针对TCDHODH活动位点半胱氨酸。预测 将评估抑制剂的抑制效力和抑制作用机制，对TCDHODH，细胞毒性 及其在细胞培养中的抗寄生作用。 X射线晶体学，片段筛选和药物化学 将合并以提供化学基础，以综合新一代有效，选择性和 药物样抑制剂。符合铅标准的前3个抑制剂（包括PK和耐受性）将具有他们的 DNDI通过对项目的实物贡献评估的功效。 该提案代表了一项空前的倡议，可显着促进建筑能力 在巴西的药物发现领域，在发展中国家的研究领域仍然非常初期。