Tofacitinib Analogs as Oxacillin Potentiators against MRSA

托法替尼类似物作为抗 MRSA 的苯唑西林增效剂

基本信息

批准号：
10607433
负责人：
Gregory Phelps
金额：
$ 4.37万
依托单位：
ST. JUDE CHILDREN'S RESEARCH HOSPITAL GRADUATE SCHOOL OF BIOMEDICAL SCIENCES, LLC
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10607433
关键词：
ADME Study Address Amines Animal Testing Anti-Bacterial Agents Anti-Infective Agents Antibiotic Resistance Antibiotics Antimicrobial Resistance Area Binding Biochemical Biological Biological Assay Biological Availability Biophysics Categories Cell physiology Chemicals Chemistry Chemosensitization Clinical Communities Complement Dangerousness Daptomycin Development Dose Drug Kinetics Drug resistance Genetic Transcription Gentian Violet Genus staphylococcus Healthcare House mice Human In Vitro Intravenous Janus kinase Lead Leadership Libraries Linezolid Luciferases Measures Mentors Methicillin Microbial Biofilms Microbiology Mind Modernization Molecular Target Oral Oxacillin Pathway interactions Pharmaceutical Preparations Phosphorylation Phosphotransferases Prevalence Prevention Prokaryotic Cells Property Proteomics Public Health Research Resistance Resources Saint Jude Children&apos s Research Hospital Sepsis Series Signal Transduction Signal Transduction Pathway Staphylococcus aureus Staphylococcus aureus infection Stomach System Systems Development Techniques Therapeutic Training Validation Vancomycin Virulence Work World Health Organization analog antimicrobial beta-Lactams biophysical analysis care burden clinical development drug discovery drug resistant pathogen efficacy study experimental study gene complementation high throughput screening in vivo in vivo evaluation interest kinase inhibitor knockout gene methicillin resistant Staphylococcus aureus microbial mortality mouse model novel pharmacologic response skills small molecule small molecule libraries transcriptomics

项目摘要

Project Summary Methicillin (oxacillin)-resistant Staphylococcus aureus (MRSA) infections are among the most frequently occurring and dangerous antibiotic-resistant public health threat. Clinical resistance to all front-line MRSA antibiotics (i.e. vancomycin, linezolid, daptomycin, and ceftaroline) has been observed, further exacerbating this threat and highlighting the urgent need for new strategies to overcome drug resistance in S. aureus. In recent years, there has been increased interest in targeting signal transduction systems for development as antimicrobials and anti-virulence agents. Targeting bacterial signal transduction is attractive because these systems are essential for many cellular processes in prokaryotes, and signal transduction pathways have been extensively studied in eukaryotic systems, as evidenced by the availability of chemical libraries containing small molecule probes with kinase inhibitor-like properties. With this strategy in mind, we screened a focused set of kinase inhibitors and identified tofacitinib, an antirheumatic Janus kinase inhibitor, as a potential chemistry starting point for the discovery of beta-lactam potentiators against MRSA. Several pyrrolopyridin-4- amine (P4A) analogs of tofacitinib were synthesized, which led to the discovery of potent analogs with the ability to re-sensitize oxacillin (OXA; a beta-lactam antibiotic) against MRSA. Preliminary pull-down proteomic experiments have identified PurM, SrrB, and PknB as putative targets, studies that have been complemented by gene knockout experiments. Therefore, the main scientific objective of this proposal is to confirm P4A engagement of putative targets using biophysical and biochemical techniques and to establish the therapeutic potential of tofacitinib analogs as anti-virulence agents and OXA potentiators against MRSA. In Aim 1, I will evaluate the binding of lead P4A compounds to putative targets, demonstrate inhibition of S. aureus kinase activity and determine the extent of potential off-target effects on mammalian kinases. Biophysical assays will be developed against the putative targets through a novel target engagement assay, which will be complemented with biochemical phosphorylation and ATP/ADP conversion assays. In Aim 2, I will evaluate the clinical potential of the P4A compounds through anti-biofilm studies, pharmacological ADME profiling, and in vivo efficacy against MRSA in a mouse model. The research plan will enhance my training in areas of target identification and validation, biophysical analysis, and anti-infective microbiology, benefiting from the expertise of my co-mentors, Drs. Richard Lee and Jason Rosch, and the state-of-the-art resources at St. Jude.

项目概要耐甲氧西林（苯唑西林）金黄色葡萄球菌 (MRSA) 感染是最常见的感染之一正在发生的危险的抗生素耐药性公共卫生威胁。对所有一线 MRSA 的临床耐药性已观察到抗生素（即万古霉素、利奈唑胺、达托霉素和头孢洛林），进一步加剧了这一威胁并强调迫切需要新的策略来克服金黄色葡萄球菌的耐药性。在近年来，人们对以信号转导系统为目标的开发越来越感兴趣抗菌剂和抗毒剂。靶向细菌信号转导很有吸引力，因为这些系统对于原核生物的许多细胞过程至关重要，并且信号转导途径已被在真核系统中进行了广泛的研究，化学文库的可用性证明了这一点具有激酶抑制剂样特性的小分子探针。考虑到这一策略，我们筛选了一个重点一组激酶抑制剂，并确定托法替尼（一种抗风湿 Janus 激酶抑制剂）作为潜在的发现抗 MRSA 的 β-内酰胺增效剂的化学起点。几种吡咯并吡啶-4- 合成了托法替布的胺 (P4A) 类似物，从而发现了有效的类似物使苯唑西林（OXA；一种 β-内酰胺抗生素）对 MRSA 重新敏感的能力。初步下拉蛋白质组学实验已确定 PurM、SrrB 和 PknB 为假定靶标，这些研究已得到补充通过基因敲除实验。因此，本提案的主要科学目标是确认P4A 使用生物物理和生化技术参与假定目标并建立托法替布类似物作为抗毒剂和 OXA 增强剂的治疗潜力耐甲氧西林金黄色葡萄球菌。在目标 1 中，我将评估先导 P4A 化合物与假定靶标的结合，证明抑制作用金黄色葡萄球菌激酶活性并确定对哺乳动物激酶的潜在脱靶影响的程度。将通过一种新颖的靶标参与测定法针对假定的靶标开发生物物理测定法，生化磷酸化和 ATP/ADP 转化检测将对其进行补充。在目标 2 中，我将通过抗生物膜研究、药理学 ADME 评估 P4A 化合物的临床潜力小鼠模型中针对 MRSA 的分析和体内功效。该研究计划将加强我的培训目标识别和验证、生物物理分析和抗感染微生物学领域，受益来自我的共同导师 Drs 的专业知识。 Richard Lee 和 Jason Rosch，以及最先进的资源圣裘德。