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

• 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
• 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' 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; 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' 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

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.

项目摘要 甲氧西林（奥沙西林） - 金黄色葡萄球菌（MRSA）感染是最常见的 发生和危险的抗生素公共卫生威胁。对所有前线MRSA的临床抵抗力 已经观察到抗生素（即万古霉素，linezolid，daptomycin和ceftaroline），进一步加剧了 这种威胁，并强调了迫切需要采取新策略来克服金黄色葡萄球菌的耐药性。在 近年来，针对以开发信号转导系统为目标的兴趣增加了 抗菌和抗病毒剂。靶向细菌信号转导很有吸引力，因为这些 系统对于原核生物中的许多细胞过程至关重要，信号转导途径一直是 在真核系统中进行了广泛的研究，这可以证明 具有激酶抑制剂样性能的小分子探针。考虑到这种策略，我们筛选了一个集中的 一组激酶抑制剂并鉴定出Tofacitinib（一种抗疾病的Janus激酶抑制剂）作为电势 针对MRSA发现β-内酰胺电位剂的化学起点。几个吡咯吡啶-4-- 合成了tofacitinib的胺（P4A）类似物，从而发现了有效的类似物 能够对MRSA重新敏感性奥沙西林（OXA；一种β-内酰胺抗生素）。初步下拉蛋白质组学 实验已将Purm，SRRB和PKNB识别为推定的目标，该研究已得到补充 通过基因敲除实验。因此，该提议的主要科学目标是确认P4A 使用生物物理和生化技术参与推定的靶标 Tofacitinib类似物的治疗潜力作为抗病毒剂和OXA增强剂的治疗潜力 MRSA。在AIM 1中，我将评估铅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的活性，并确定潜在的脱靶对哺乳动物激酶的影响的程度。 通过新的目标参与测定法，将针对推定的目标开发生物物理测定法， 它将与生化磷酸化和ATP/ADP转换测定法相辅相成。在AIM 2中，我 将通过抗生物膜研究评估P4A化合物的临床潜力 在小鼠模型中对MRSA进行分析和体内功效。研究计划将增强我的培训 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 从我的联合给予者的专业知识中。理查德·李（Richard Lee）和杰森·罗奇（Jason Rosch），以及最先进的资源 圣裘德。