Selective inhibitors of phosphoglycerate mutase and phosphoglycerate kinase as le

磷酸甘油酸变位酶和磷酸甘油酸激酶的选择性抑制剂如

• 批准号: 8069449
• 负责人: Malcolm Douglas Walkinshaw
• 金额: $ 2.7万
• 依托单位: UNIVERSITY OF EDINBURGH
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-12-17 至 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8069449
• 关键词: Africa; Africa South of the Sahara; African Trypanosomiasis; Asia; Binding Sites; Biological Assay; Calorimetry; Cells; Central America; Cessation of life; Chagas Disease; Chemicals; Consumption; Country; Couples; Crystallization; Crystallography; Databases; Detection; Development; Disease; Disease Resistance; Drug Delivery Systems; Economic Burden; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzymes; Equilibrium; Firefly Luciferases; Glycolysis; Goals; Growth; Human; Institutes; Kinetics; Lead; Leishmania; Ligands; Measurement; Metabolic; Methods; Mining; Molecular Bank; Molecular Biology; Molecular Models; NADH; Nature; Parasites; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phosphoglycerate Kinase; Phosphoglycerate Mutase; Phosphotransferases; Population; Poverty; Procedures; Protein Isoforms; Proteins; Public Health; Pyruvate Kinase; RNA Interference; Reaction; Reading; Research; Resistance; Risk; Roentgen Rays; Screening procedure; Series; Site-Directed Mutagenesis; South America; Specificity; Staging; Structure; Structure-Activity Relationship; Surface Plasmon Resonance; Testing; Titrations; Toxic effect; Trypanosoma brucei brucei; Trypanosoma cruzi; Universities; Validation; Visceral Leishmaniasis; analog; base; chemical synthesis; design; enolase; enzyme activity; health economics; high throughput screening; human disease; improved; inhibitor/antagonist; interest; light scattering; molecular modeling; novel; novel strategies; nutrition; pathogen; preclinical study; programs; relational database; repository; response; scaffold; small molecule; structural biology; three dimensional structure; virtual

DESCRIPTION (provided by applicant): The goal of this project is to discover and optimize selective inhibitors of phosphoglycerate mutase (PGAM) and phosphoglycerate kinase (PGK) of trypanosomatid parasites (Trypanosoma brucei, T. cruzi and Leishmania species). These pathogens cause serious, often fatal diseases of humans such as sleeping sickness, Chagas' disease and kala-azar in tropical and subtropical countries primarily in Africa, Central and South America, and Asia where over 30 million people are infected, with approximately 130,000 deaths annually. Tragically, current drugs for their treatment are unsatisfactory because they are toxic and ineffective against some forms of the diseases, and resistance is becoming increasingly common. Glycolysis is essential in the infective stage of T. brucei (Tb) and therefore a promising drug target. Inhibitors of glycolytic enzymes such as PGAM and PGK may thus serve as lead compounds for the development of new drugs. PGAM is a particularly attractive target because the corresponding host enzyme is not homologous and has no features in common with the parasite enzyme. TbPGAM has been validated as a drug target by RNAi, and TbPGK by inhibitor studies. Detailed structural information is already available for both enzymes. The proposed research has as specific aims: (1) To develop and validate novel forward and reverse assays mounted on the NCGC high-throughput screening platform for trypanosomatid PGAM and PGK. The four assays will serve as both primary and orthogonal assays for confirmation of actives. (2) To use the four assays to screen the Molecular Library Small Molecule Repository (MLSMR) containing more than 300,000 small molecules. Unique structural features of trypanosomatid PGAM and PGK (compared with the corresponding human enzymes) will be exploited for the discovery of selective inhibitors of these enzymes initially from T. brucei and L. Mexicana. (3) To compare sets of actives in the forward and reverse directions. Preliminary assays on TbPGK have already shown the possibility to obtain inhibitors and activators of different chemotypes when assayed in the two directions. (4) To confirm the potency of these compounds in a panel of secondary hit validation assays and to test their specificity in kinetic and selectivity assays, and to further improve the potency of the most promising molecules thus obtained by structure-based methods, analogue synthesis and medicinal chemical principles. (5) To determine the mode of action of the most promising molecules by enzyme assays, protein crystallography and biophysical measurements. (6) To test compounds displaying the highest potency for their ability to inhibit growth of cultured trypanosomatid cells representing pathogenic stages of the parasites, as well as for lack of toxicity toward cultured human cells. PUBLIC HEALTH RELEVANCE: Sleeping sickness in sub-Saharan Africa, Chagas' disease in Central and South America, and kala-azar and related diseases in tropical and subtropical regions of the world cause severe public health and economic burdens on populations that are already caught in a tragic cycle of poverty, poor nutrition and disease. Millions of people worldwide are infected by these potentially fatal diseases and hundreds of millions are at risk. Existing treatments have developed little in the past 50 years, and suffer from toxicity, inefficiency and resistance; the goal of this project is to develop lead drugs that will be suitable for entry into pre-clinical trials.

描述（由申请人提供）：本项目的目的是发现和优化锥虫寄生虫（布氏锥虫，T. cruzi和利什曼原虫物种）。这些病原体在热带和亚热带国家（主要是非洲、中美洲和南美洲以及亚洲）引起严重的、往往是致命的人类疾病，例如嗜睡病、恰加斯病和黑热病，这些国家有超过3000万人感染，每年约有130，000人死亡。不幸的是，目前用于治疗它们的药物不能令人满意，因为它们对某些形式的疾病是有毒的和无效的，并且耐药性变得越来越普遍。糖酵解是T.布氏杆菌（Tb），因此是有希望的药物靶点。因此，糖酵解酶抑制剂如PGAM和PGK可作为开发新药的先导化合物。PGAM是一个特别有吸引力的靶标，因为相应的宿主酶不是同源的，并且与寄生虫酶没有共同的特征。TbPGAM已被RNAi验证为药物靶标，TbPGK已被抑制剂研究验证为药物靶标。这两种酶的详细结构信息已经可以获得。本研究的主要目的是：（1）建立和验证新型的锥虫PGAM和PGK的正向和反向检测方法。这四种试验将作为确认活性物质的主要和正交试验。(2)使用四种检测方法筛选包含超过30万个小分子的分子库小分子库（MLSMR）。锥虫PGAM和PGK的独特结构特征（与相应的人类酶相比）将被用于发现这些酶的选择性抑制剂。brucei和L.墨西哥(3)在正向和反向方向上比较活性物组。对TbPGK的初步测定已经显示了当在两个方向上测定时获得不同化学型的抑制剂和激活剂的可能性。(4)在一组二次命中验证试验中确认这些化合物的效价，并在动力学和选择性试验中检测其特异性，并进一步提高通过基于结构的方法、类似物合成和药物化学原理获得的最有前途的分子的效价。(5)通过酶分析、蛋白质晶体学和生物物理测量确定最有希望的分子的作用模式。(6)为了测试化合物显示出抑制代表寄生虫致病阶段的培养锥虫细胞生长的能力以及对培养的人细胞无毒性的最高效力。 公共卫生关系：撒哈拉以南非洲的昏睡病、中美洲和南美洲的恰加斯病以及世界热带和亚热带地区的黑热病和相关疾病给已经陷入贫困、营养不良和疾病悲惨循环的人口造成了严重的公共卫生和经济负担。全世界有数百万人感染了这些可能致命的疾病，数亿人处于危险之中。现有的治疗方法在过去的50年里几乎没有发展，并且存在毒性，效率低下和耐药性;该项目的目标是开发适合进入临床前试验的先导药物。