Inhibitors of CoA biosynthesis as novel antitubercular and antistaphylococcal agents

作为新型抗结核和抗葡萄球菌药物的 CoA 生物合成抑制剂

基本信息

批准号：
10113512
负责人：
Erick Strauss
金额：
$ 13.1万
依托单位：
STELLENBOSCH UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-03-05 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10113512
关键词：
Address Adherence Aerobic Anabolism Anaerobic Bacteria Anti-Bacterial Agents Antibiotics Antitubercular Agents Back Bacteria Binding Biochemical Biological Assay Bypass Cells Cellular Assay Clinical Coenzyme A Communicable Diseases Communities Cytosine Data Developing Countries Development Doctor of Philosophy Drug Design Drug Targeting Drug resistance Drug resistance in tuberculosis Energy Metabolism Energy-Generating Resources Enzymes Evaluation Fermentation Foundations Genetic study Goals Growth HIV Health Hospitals Human In Situ In Vitro Infection Infectious Agent International Life Measures Metabolic Activation Metabolism Modeling Mus Mycobacterium tuberculosis Mycobacterium tuberculosis H37Rv Natural Products Nosocomial Infections Organism Pathway interactions Physiological Prevalence Process Prodrugs Proteins Reaction Resistance Respiration Ribose Series South Africa Specificity Staphylococcus aureus Structure Structure-Activity Relationship System Testing Therapeutic Agents Tuberculosis Universities Virulence analog bactericide base cell envelope chronic infection co-infection cofactor cytotoxicity design enzyme activity enzyme pathway experimental study fatty acid biosynthesis genetic resistance improved in vivo inhibitor/antagonist interest macrophage mutant novel pathogen phosphonate prevent priority pathogen small molecule small molecule inhibitor small molecule therapeutics targeted agent tuberculosis treatment

项目摘要

PROJECT SUMMARY/ABSTRACT Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), continues to be one of the world’s deadliest infectious diseases because of several factors, including a) poor adherence to the long-term multidrug therapy, b) latent (or persistent) infection, c) co-infection with HIV and d) drug-resistant Mtb strains that are unaffected by current TB therapies. Similarly, Staphylococcus aureus was recently listed by the WHO as a “high priority” pathogen in regards to new antibiotic development, due to its increasing prevalence as a major infectious agent both in hospitals and in the community, and to increased virulence and rising resistance against current antistaphylococcal agents. The biosynthesis of coenzyme A (CoA), an essential cofactor relevant to several metabolic processes, contains an emerging set of targets for antibacterial drug design. Small molecule inhibitors of this pathway have shown activity in a cell-based model of latent TB, increasing interest in this set of targets as a potential means attacking the latent pathogen. Similarly, CoA biosynthesis in S. aureus has recently been associated with maintaining the organism under anaerobic conditions, a model for persistent infections. The broad, long-term objective of this project is to develop inhibitors of CoA biosynthesis, which act on the PPCS enzyme activity of Mtb and S. aureus, that show whole cell activity under a range of conditions and potential for further development as clinically useful agents that could be used in combination treatments. This goal has strong foundations in results from in vitro genetic studies demonstrating the vulnerability of the target, and in vivo experiments showing that depletion of PPCS activity is bactericidal and prevents a robust Mtb infection in mice. In S. aureus, a natural product that targets PPCS activity shows highly selective whole cell activity. These results suggest that the PPCS activity is a highly promising target that is tractable to inhibition by small molecules. To develop new whole cell active compounds, we are proposing inhibitor structures based on a well-established strategy that has successfully been used to prepare inhibitors of mechanistically similar enzymes, giving rise to compounds that also showed excellent whole cell activity against Mtb and S. aureus. In addition, we are proposing complementary metabolic activation and prodrug approaches to address the known challenge of preparing compounds that are able to cross the Mtb cell envelope. We will pursue this objective through three Specific Aims: 1) Design and synthesis of Pan-CMP mimics as PPCS inhibitors; 2) Biochemical evaluation of metabolic activation and PPCS inhibition of Aim 1 compounds; and 3) Evaluation of PPCS inhibitors in models of actively growing and persistent Mtb and S. aureus. These aims are formulated to systematically optimize the structure of the inhibitors and to follow their on-target activity and selectivity through enzymatic and whole cell assays, the latter including tests against wild-type Mtb and S. aureus and mutant Mtb strains (to show target specificity), models of latent/persistent growth for both Mtb and S. aureus, and an ex vivo macrophage model (in the case of Mtb).

项目概要/摘要结核病 (TB) 由结核分枝杆菌 (Mtb) 引起，仍然是世界上最严重的疾病之一由于多种因素，这些疾病成为最致命的传染病，包括：a) 长期坚持不力多种药物治疗，b) 潜伏（或持续）感染，c) HIV 合并感染，d) 耐药 Mtb 菌株同样，金黄色葡萄球菌最近被世界卫生组织列入名单。作为新抗生素开发的“高度优先”病原体，因为它作为一种细菌的流行率不断增加医院和社区中的主要传染源，并导致毒力增加和抵抗力增强对抗当前的抗葡萄球菌药物。辅酶 A (CoA)（一种重要的辅助因子）的生物合成。与多种代谢过程相关，包含一组新兴的抗菌药物设计目标。该途径的小分子抑制剂已在基于细胞的潜伏性结核病模型中显示出活性，增加了对这组目标的兴趣作为攻击潜在病原体的潜在手段，同样，CoA 生物合成。最近，金黄色葡萄球菌与在厌氧条件下维持生物体有关，这是一种模型该项目的广泛、长期目标是开发 CoA 生物合成抑制剂，作用于 Mtb 和 S. aureus 的 PPCS 酶活性，在一定范围内显示出全细胞活性条件和进一步开发作为可联合使用的临床有用药物的潜力这一目标以体外遗传学研究的结果为基础，证明了这一点。靶标的脆弱性，体内实验表明，PPCS 活性的耗尽具有杀菌作用在金黄色葡萄球菌中，一种针对 PPCS 活性的天然产物显示出高度的预防 Mtb 感染的能力。这些结果表明 PPCS 活性是一个非常有前途的目标。为了开发新的全细胞活性化合物，我们建议。基于已成功用于制备抑制剂的成熟策略的抑制剂结构机制相似的酶，产生的化合物也表现出优异的全细胞活性此外，我们还建议补充代谢激活和前药。解决制备能够穿过 Mtb 细胞的化合物的已知挑战的方法我们将通过三个具体目标来实现这一目标：1）Pan-CMP 的设计和合成。模拟作为 PPCS 抑制剂；2) 目标 1 的代谢激活和 PPCS 抑制的生化评估 3) 在活跃生长和持久性 Mtb 和 S 模型中评估 PPCS 抑制剂。制定这些目标是为了系统地优化抑制剂的结构并遵循它们的要求。通过酶促和全细胞测定来确定目标活性和选择性，后者包括针对野生型 Mtb 和金黄色葡萄球菌以及突变型 Mtb 菌株（以显示靶标特异性）、潜伏/持续模型 Mtb 和金黄色葡萄球菌的生长，以及离体巨噬细胞模型（以 Mtb 为例）。