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: 10297860
• 负责人: Zhuo Michael Wang
• 金额: $ 47.68万
• 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 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10297860
• 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; Modeling; Organ; Organism; Outcome; Parasites; Parasitic Diseases; Pathogenicity; Pathway interactions; Pharmaceutical Preparations; Phlebotominae; Prodrugs; Proteins; Public Health; Reaction; Resistance; Roentgen Rays; Role; Route; Skin; Sterols; Structure; Testing; Vaccines; Visceral Leishmaniasis; assay development; base; chemotherapy; cost; cytotoxicity; drug discovery; effective therapy; high throughput screening; inhibitor; 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.

概括 人类利什曼病是一种由属于原生动物的寄生虫引起的毁灭性传染病 利什曼尼亚属。该疾病在90多个国家 /地区发现，估计为1-2 每年在全球范围内有百万个新感染。利什曼病在返回美国军事人员方面也很常见 从伊拉克和阿富汗出发，自2001年以来报告了2000多起病例。 导致皮肤疮（皮肤利什曼病或CL）以及威胁生命的内部感染 器官（内脏利什曼病或VL）。 VL（所有利什曼病病例的20％）是最危险和致命的 疾病的形式，如果未经治疗，死亡率接近100％。这是第二大致命的寄生虫 世界上的疾病（疟疾之后）。尽管在许多内在地区是一个严重的公共卫生问题，但 没有可用于控制利什曼病的疫苗或预防化疗。当前的 抗精神病药的有效性有限，严重的副作用和高成本。因此，有一个重大的未满足 医疗需求对利什曼病的安全有效药物。 Leishmanial CYP5122A1是一部小说 细胞色素P450（CYP）酶，这对于Donovani L. donovani的生存至关重要，Donovani是一种主要的病因。 VL。独立和我们自己的研究使我们假设选择性CYP5122A1的识别 抑制剂作为化学问题将允许通过 利什曼尼亚。为了检验我们的假设，提出了三个具体目标来回答三个主要问题：1） 在麦角固醇生物合成途径中比较CYP5122A1和CYP51的生化作用 利什曼尼亚？ 2）可以识别和验证选择性CYP5122A1抑制剂吗？和3）是芳基酰亚胺 （AIAS）和/或AIA-唑杂种潜在的化学问题，以帮助划定CYP5122A1的生化作用 创新的实验方法将在拟议的项目中进行，例如 新的基于HPLC/MS的固醇测定法，基于荧光的CYP5122A1和CYP51抑制测定法，以及 尖端的高内容成像基于细胞内的抗精神病药物评估。如果成功，我们的建议 鉴定新型CYP5122A1抑制剂的项目将阐明该蛋白在利什曼原虫生物学和 将CYP5122A1验证为药物靶标。这样的结果可能会对长期产生重大积极影响 利什曼病在全球范围内控制。

项目成果

Deciphering the Molecular Mechanism and Function of Pore-Forming Toxins using Leishmania major.

• DOI: 10.3791/64341
• 发表时间: 2022-10-28
• 期刊: Journal of visualized experiments : JoVE
• 作者: Haram CS;Moitra S;Keane R;Breslav E;Zhang K;Keyel PA
• 通讯作者: Keyel PA

Synthesis and Antileishmanial Evaluation of Arylimidamide-Azole Hybrids Containing a Phenoxyalkyl Linker.

• DOI: 10.1021/acsinfecdis.0c00855
• 发表时间: 2021-07-09
• 期刊: ACS infectious diseases
• 影响因子: 5.3
• 作者: Abdelhameed A;Feng M;Joice AC;Zywot EM;Jin Y;La Rosa C;Liao X;Meeds HL;Kim Y;Li J;McElroy CA;Wang MZ;Werbovetz KA
• 通讯作者: Werbovetz KA

Sterol profiling of Leishmania parasites using a new HPLC-tandem mass spectrometry-based method and antifungal azoles as chemical probes reveals a key intermediate sterol that supports a branched ergosterol biosynthetic pathway.

• DOI: 10.1016/j.ijpddr.2022.07.003
• 发表时间: 2022-12
• 期刊: INTERNATIONAL JOURNAL FOR PARASITOLOGY-DRUGS AND DRUG RESISTANCE
• 影响因子: 4
• 作者: Feng, Mei;Jin, Yiru;Yang, Sihyung;Joachim, Arline M.;Ning, Yu;Mori-Quiroz, Luis M.;Fromm, Jacob;Perera, Chamani;Zhang, Kai;Werbovetz, Karl A.;Wang, Michael Zhuo
• 通讯作者: Wang, Michael Zhuo

Balancing de novo synthesis and salvage of lipids by Leishmania amastigotes.

• DOI: 10.1016/j.mib.2021.07.004
• 发表时间: 2021-10
• 期刊: Current opinion in microbiology
• 影响因子: 5.4
• 作者: Zhang K

CYP5122A1 encodes an essential sterol C4-methyl oxidase in Leishmania donovani and determines the antileishmanial activity of antifungal azoles.

CYP5122A1 编码杜氏利什曼原虫中必需的甾醇 C4-甲基氧化酶，并决定抗真菌唑类的抗利什曼活性。

• DOI: 10.21203/rs.3.rs-3185204/v1
• 发表时间: 2023
• 期刊: Research square
• 作者: Jin,Yiru;Basu,Somrita;Feng,Mei;Ning,Yu;Munasinghe,Indeewara;Joachim,ArlineM;Li,Junan;Madden,Robert;Burks,Hannah;Gao,Philip;Perera,Chamani;Werbovetz,KarlA;Zhang,Kai;Wang,MichaelZhuo
• 通讯作者: Wang,MichaelZhuo