Development of WecA-targeting immune potentiators to treat carbapenem-resistant Enterobacteriaceae (CRE) infections

开发 WecA 靶向免疫增强剂来治疗碳青霉烯类耐药肠杆菌 (CRE) 感染

基本信息

批准号：
10415522
负责人：
Terry Roemer
金额：
$ 100万
依托单位：
PROKARYOTICS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10415522
关键词：
Address Anabolism Antibiotics Bacteremia Bacteria Bacterial Infections Bacterial Pneumonia Binding Cancerous Carbapenems Cell Survival Cells Centers for Disease Control and Prevention (U.S.)Chemicals Clinical Colistin Communities Complement Data Development Dose Enterobacter Enterobacteriaceae Enterobacteriaceae Infections Epidemic Escherichia coli Essential Genes Exposure to Formulation Fosfomycin Genes Genetic Goals Gram-Negative Bacteria Growth HepG2 Hospitals Imipenem Immune Immune Targeting Immune system Immunooncology In Vitro Infection Innate Immune System Ion Channel Klebsiella pneumoniae Lead Libraries Life Maps Mediating Meropenem Modeling Multi-Drug Resistance Mutation New Agents O Antigens Oral Organism Orthologous Gene Pathogenesis Pathway interactions Pattern Permeability Pharmaceutical Preparations Pharmacology Phase Plasmids Polymyxins Production Property Pseudomonas aeruginosa Rattus Reporter Reporting Resistance Risk Rodent Rodent Model Route Safety Septicemia Serum Siderophores Source Superbug Taro Vegetable Testing analog antimicrobial bactericide base beta-Lactam Resistance beta-Lactamase beta-Lactams carbapenem resistance carbapenem-resistant Enterobacteriaceae commensal bacteria doripenem drug repurposing extracellular gene product gut microbiota in vivo inhibitor/antagonist innovation member mortality mutant new therapeutic target novel novel therapeutics overexpression pathogen periplasm programmed cell death protein 1 public health relevance resistance mechanism scale up screening success tigecycline

项目摘要

Abstract In 2013, the CDC designated carbapenem-resistant Enterobacteriaceae (CRE) an Urgent Threat, and in 2017, the WHO designated it a Priority 1 “critical superbug”. As few therapies remain to treat CRE, the risk of “pan- resistant” CRE, untreatable by any currently available antibiotic, increases. Entirely new agents with novel mechanisms of action (MOA) not cross-resistant to SOC agents languish. Our proposal aims to develop an O- antigen (O-a) biosynthetic inhibitory agent that potentiates serum-mediated killing (SMK) and is efficacious in a CRE rodent model of infection. We have shown that the O-a biosynthesis gene wecA, a nonessential gene under standard growth conditions, is essential for growth and pathogenesis in the presence of mammalian serum. Our proposal outlines a plan to develop synthetic inhibitors of WecA that we previously discovered and optimized to inhibit the Gram-positive WecA ortholog, TarO. Our Aims are: Aim 1 (Phase 1; Ph1). Screening, MOA studies, and Proof-of-Concept in vivo Studies. (1) Complete lux reporter screening of the tarocin focused library for SMK against WT and ΔtolC E. coli (Ec), (2) directly confirm tarocins inhibit EcWecA as their MOA for eliciting SMK, (3) demonstrate that tarocin-induced SMK extends to Kp, and (4) demonstrate proof-of-concept in vivo efficacy. Milestone 1. Screen ~600 additional tarocin analogs for SMK activity and identify up to 4 chemically distinct tarocin subseries demonstrating i) > 4-fold EC50 shift in SMK by EcWecA overexpression, ii) dose-dependent depletion of O-a in a whole-cell context, iii) causal drugR mutations mapping to EcwecA, iv) SMK activity against Kp DtolC, v) >90% HepG2 cell viability at 25X MIC, and vi) favorable 50% protective dose for survival in a rat septicemia model using an efflux-deficient Ec strain. Aim 2 (Ph2). Lead ID/Opt. Identify tarocins with potent WT Ec and Kp SMK by (1) empirically testing analogs for SMK against panel of Ec/Kp permeability/efflux deficient mutants, (2) employing recent physicochemical rules of GN entry, and (3) exploring siderophore conjugation to drive SAR efforts. We will also optimize PK and drug- like properties. Milestone 2. Identify up to 3 analogs demonstrating i) Ec/Kp WT activity (MIC in serum [MICs] <1 ug/ml), ii) PK exposure to cover MICs for 4 hrs, iii) Ec/Kp target/pathway engagement selectivity including tarocinR wecA mutation in Kp, v) Ec/Kp FOR <1x10-9 at 8X MICs, vi) MICs90 <2 ug/ml (100 isolates), and vii) > 90% HepG2/HEK293 viability at 50X MICs, clean vs CYP/ion channels (>10 uM), and PanLabs IC50 >10 uM. Aim 3 (Ph2). In vivo efficacy demonstration. (1) Optimize synthetic routes to efficiently prepare 3 analogs for formulation and dose-ranging rat PK studies, (2) identify formulation vehicles for the 3 analogs to enable oral and IV PK dosing in in vivo studies, and (3) demonstrate in vivo efficacy for our top compound in a rodent septicemia model using Ec and Kp strains. Milestone 3. Synthesize 500 mg (>95% purity) of up to 3 optimal analogs that best satisfy Aim 2 milestones, identify formulation in a safety approved vehicle that achieves 10X MICS90 target exposure, and demonstrate > 3 log reduction of Ec and Kp bacterial burden after 24h treatment.

抽象的 2013年，疾病预防控制中心（CDC）指定了耐碳青霉烯（CRE）的紧急威胁。由于仍然很少有治疗CRE的疗法，因此任何当前可用的抗生素都无法治疗的“抗泛抗” CRE的风险会增加。具有新颖的作用机制（MOA）的全新代理商（MOA）对SOC代理商的耐药性不稳定。我们的建议旨在开发WECA的O-抗原（O-A）生物合成抑制剂，我们先前发现并优化以抑制O-A生物合成基因WECA，这是一种非必需的基因WECA，这是一种非必需的标准基因，这是哺乳动物血清的生长和发病率的必不可少的。我们的提案概述了开发WECA合成抑制剂的计划，我们以前发现并优化了以抑制革兰氏阳性的WECA直系同源物，Taro。我们的目标是：目标1（阶段1; PH1）。筛查，MOA研究和体内研究证明。（1）完整的Lux记者筛选了针对WT和ΔTOLCE. coli（EC）的Tarocin集中库，（2）直接确认Tarocins抑制ECWECA作为其引起SMK的MOA，（3）证明了Tarocin诱导的SMK延伸至KP，并证明了（4）证明了Vivo in Vivo invivo invivo inconcon in vivo inconcon in vivo inconcon in vivo inconcon in vivo expeipy。里程碑1。筛查〜600个用于SMK活性的额外的塔罗蛋白类似物，并确定多达4个不同的tarocin tarocin subsies i）通过ECWECA通过ECWECA过表达来证明i）> 4倍EC50的EC50偏移，ii）O-A的剂量依赖性依赖性依赖于全细胞的剂量依赖性依赖于全细胞的依赖性，iii II I III中的Causal药物反对ECWECA的活动，IV）Smkap Apptivation kp Aptive kp dt> kp dt）kp dt）在25倍MIC处的HEPG2细胞活力，VI）使用排出不足的EC菌株在大鼠败血症模型中可保护50％的剂量。 AIM 2（PH2）。线索ID/OPT。通过（1）通过（1）对SMK进行EC/KP渗透性/外排的缺陷突变体的SMK进行验证，（1）通过（1）通过（1）鉴定塔罗蛋白，（2）采用最近的GN进入物理规则，以及（3）探索siderophore驱动SAR努力的siderophore。我们还将优化PK和类似药物的特性。里程碑2。确定多达3个类似物，证明I）EC/KP WT活性（麦克风[MICS] <1 ug/ml），ii）PK暴露于4小时的MIC，III）EC/KP目标/途径参与性选择性，包括<1x10-MIC 9 ug 1x10-mIC，包括<1x10-9，包括<1x10-9 （100个分离株）和vii）> 90％HEPG2/HEK293在50x MICS，清洁与CYP/离子通道（> 10 UM）和Panlabs IC50> 10 um。目标3（ph2）。体内效率示范。（1）优化合成途径，以有效准备3个类似物，以进行配方奶粉和剂量范围的大鼠PK研究，（2）确定3种类似物的配方奶粉工具以在体内研究中启用口服和IV PK剂量，（3）使用Rodent Spescound使用EC和Kp strains在体内效率。里程碑3。合成500毫克（> 95％纯度）的最多3个最佳类似物，最能满足AIM 2里程碑，在安全批准的车辆中识别出可实现10倍MICS90目标暴露的车辆中的公式，并在24H治疗后证明了> 3对EC和KP细菌燃烧的原木减少。