Development of a mechanistically novel Gram-negative antibiotic targeting MsbA-mediated Lipopolysaccharide Biogenesis

开发一种机制新颖的革兰氏阴性抗生素，靶向 MsbA 介导的脂多糖生物发生

• 批准号: 10584170
• 负责人: Terry Roemer
• 金额: $ 67.93万
• 依托单位: PROKARYOTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-11-04 至 2027-10-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10584170
• 关键词: ATP-Binding Cassette Transporters; Address; American; Anti-Bacterial Agents; Antibiotic susceptibility; Antibiotics; Bacterial Infections; Binding Proteins; Biochemical; Biogenesis; Cell Survival; Cells; Chemicals; Clinical; Coculture Techniques; Communities; Complex; Cytoplasm; Development; Dose; Drug Design; ESKAPE pathogens; Economic Burden; Enterobacteriaceae; Enterobacteriaceae Infections; Enzymes; Equation; Escherichia coli; Escherichia coli Infections; European; Extended-spectrum β-lactamase; Formulation; Fright; Goals; HepG2; Hepatocyte; Hospitalization; Hospitals; In Vitro; Incidence; Infection; Ion Channel; Kidney; Klebsiella pneumoniae; Lactamase; Lactams; Lead; Left; Life; Lipid A; Lipopolysaccharides; Marketing; Measures; Mediating; Membrane; Membrane Proteins; Microsomes; Modeling; Monitor; Multi-Drug Resistance; Mus; New Agents; Nosocomial Infections; O Antigens; Outpatients; Pathway interactions; Patients; Peritonitis; Pharmaceutical Preparations; Phase; Plasma; Plasma Proteins; Property; Resistance; Resolution; Roentgen Rays; Series; Structure; Structure-Activity Relationship; Superbug; Testing; Therapeutic; Time; Urinary tract infection; acute pyelonephritis; analog; bactericide; carbapenem resistance; carbapenem-resistant Enterobacteriaceae; clinical development; community setting; cost; cytotoxicity; fluoroquinolone resistance; improved; in vitro testing; in vivo; indexing; inhibitor; innovation; lead optimization; lead series; meetings; membrane biogenesis; minimal inhibitory concentration; novel; overexpression; pathogen; periplasm; pharmacokinetics and pharmacodynamics; pre-clinical; programs; resistance frequency; safety study; scale up; small molecule; small molecule inhibitor; subcutaneous; synergism; trend

Recently, WHO and CDC designated Carbapenem-resistant Enterobacteriaceae (CRE) a Priority 1 ‘critical superbug’ and an ‘Urgent Threat’, and warned that new treatments for superbugs, which kill nearly 50,000 Americans and Europeans a year, are unlikely to be developed in time if left to market forces alone. Few therapeutic options are left to treat CRE, and the fear of ‘pan-resistant’ CRE has emerged. Currently, most CRE infections occur in a hospital setting, but the potential spread of CRE in the community also exists. Entirely new agents with novel mechanisms of action languish; completely novel antibacterials with a new mechanism of action and lacking clinical cross-resistance to existing drug classes are urgently needed. Our proposal aims to develop a mechanistically novel, IV and PO administered agent to treat uncomplicated and complicated UTI cases caused by antibiotic susceptible and multidrug-resistant Enterobacteriaceae, including extended spectrum beta-lactamase producers and CRE. Using an innovative overexpression-based co-culture screen in Escherichia coli (Ec), we identified four structurally distinct series of small molecule inhibitors targeting MsbA, an essential and broadly conserved Gram-negative (GN) ABC transporter responsible for lipopolysaccharide (LPS) biogenesis and construction of the Gram-negative outer membrane (OM). A prioritization of the four series through our current R21AI146541-supported program has enabled selection of a series to enter the Lead Identification (ID) phase of development. We propose the following aims to develop this series: Aim 1 (2 years) - Lead ID to select a single lead series with WT activity and efficacy for Aim 2 Lead Optimization (Op): 1.1) exploratory med chem to improve WT activity, 1.2) obtain high resolution MsbAi-MsbA X-ray co-crystal structure to guide structure-based drug design (SBBD) using the Schrodinger Discover Suite, 1.3) monitor whole cell (MIC) activity, 1.4) track in vitro IC50 potency, 1.5) test ppb, mammalian cytotoxicity, and P-gp inhibition/stimulation, 1.6) FOR, time kill curves, and LpxC inhibitor synergy studies, 1.7) measure PK, 1.8) determine Enterobacteriaceae MIC90, 1.9) synthetic scale up, 1.10) dose-ranging mouse PK/formulation studies, and 1.11) demonstrate in vivo efficacy in a mouse peritonitis model of WT Ec infection. Aim 2 (3 years) - Lead Op to prepare a Pre-Clinical Candidate (PCC) for subsequent safety studies: 2.1) med chem optimization of drug-like properties, 2.2) track MIC, IC50, and cytotoxicity, 2.3) conduct in vitro ADME, 2.4) single mouse (IV, IP, PO) PK, 2.5) validate MOA and FOR, 2.6) test in vitro mammalian off target-activity, 2.7) expanded Enterobacteriaceae MIC90, 2.8) dose-ranging mouse PK, 2.9) synthetic scale up, 2.10) efficacy in murine UTI model, 2.11) identify PK/PD index, 2.12) expanded off target-activity as in 2.6.

最近，世界卫生组织和疾病预防控制中心将耐碳青霉烯类肠杆菌科(CRE)列为优先1‘关键 并警告说，针对导致近5万人死亡的超级细菌的新疗法 美国人和欧洲人一年来，如果单靠市场力量，不太可能及时发展起来。一些 治疗CRE的选择剩下了，人们已经出现了对“泛耐药”CRE的恐惧。目前，大多数CRE 感染发生在医院环境中，但社区中也存在CRE的潜在传播。全新的 具有新作用机制的药物萎靡不振；具有新作用机制的全新抗菌药 对现有药物类别缺乏临床交叉抗药性是迫切需要的。我们的建议旨在 开发一种治疗简单和复杂尿路感染的机械新型静脉注射和口服给药药物 由抗生素敏感和多重耐药肠杆菌科引起的病例，包括超广谱 产β-内酰胺酶和CRE。在大肠杆菌中使用创新的基于过度表达的共培养筛选 Coli(EC)，我们确定了四个结构上不同的小分子抑制剂系列，靶向MSBA，一个必不可少的 和广泛保守的负责内毒素的革兰氏阴性(GN)ABC转运体 革兰氏阴性杆菌外膜的生物发生和构建四个系列的优先顺序 通过我们当前的R21AI146541支持的计划，我们能够选择一个系列进入领先地位 开发的标识(ID)阶段。我们提出以下目标来开发本系列： 目标1(2年)-为目标2选择具有WT活性和有效性的单个导联ID 优化(Op)：1.1)探索改善WT活性的方法，1.2)获得高分辨率的MsbAi-MSBA 使用薛定谔发现套件的X射线共晶体结构来指导基于结构的药物设计(SBBD)， 1.3)监测全细胞(MIC)活性，1.4)跟踪体外IC50效力，1.5)测试ppb，哺乳动物细胞毒性，以及 P-gp抑制/刺激，1.6)时间杀伤曲线，以及LpxC抑制剂协同研究，1.7)测量pk，1.8) 确定肠杆菌科MIC90，1.9)合成放大，1.10)剂量范围小鼠PK/配方研究， 和1.11)在WT EC感染的小鼠腹膜炎模型中展示了体内疗效。 目标2(3年)-领导运营人员为后续安全性研究准备临床前候选人(PCC)：2.1) 类药物性质的MED化学优化，2.2)跟踪MIC、IC50和细胞毒性，2.3)进行体外ADME， 2.4)单个小鼠(IV，IP，PO)PK，2.5)验证MOA和FOR，2.6)体外哺乳动物脱靶活性测试， 2.7)扩大的肠杆菌科MIC90，2.8)剂量范围的小鼠PK，2.9)合成放大，2.10)疗效 在小鼠尿路感染模型中，2.11)确定PK/PD指数，2.12)扩大目标活动，如2.6。