Host defense-stimulating macrocyclic peptides for treatment of MDR bacterial infections

用于治疗耐多药细菌感染的宿主防御刺激大环肽

• 批准号: 9145465
• 负责人: Andre J. Ouellette
• 金额: $ 149.57万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9145465
• 关键词: ADME Study; Acute Toxicity Tests; Animal Model; Antibiotic Resistance; Antibiotics; Attenuated; Bacteria; Bacterial Infections; Biochemical; Biological Markers; Carbapenems; Characteristics; Coagulation Process; Collaborations; Complement; Complex; Cyclic GMP; Cyclic Peptides; Data; Defensins; Development; Dose; Drug Kinetics; Engineering; Enterobacteriaceae; Enterobacteriaceae Infections; Epidemic; Escherichia coli; Evaluation; Excretory function; Goals; Hand; Host Defense; Immune; Immune response; Immune system; Immunity; In Vitro; Inbred BALB C Mice; Infection; Injectable; Kilogram; Kinetics; Klebsiella; Klebsiella pneumonia bacterium; Knowledge; Label; Lactamase; Lead; Leukocytes; Maximum Tolerated Dose; Mediating; Mediator of activation protein; Membrane; Metabolism; Methods; Modeling; Multi-Drug Resistance; Mus; NZW Mouse; Oryctolagus cuniculus; Pan Genus; Penicillin-Binding Proteins; Peptides; Pharmaceutical Preparations; Pharmacodynamics; Pharmacologic Substance; Plant Leaves; Plants; Production; Property; Public Health; Publishing; Rattus; Recombinants; Reporting; Research; Resistance; Resistance profile; Resort; Resources; Selection Criteria; Sepsis; Series; Solubility; Solvents; System; Technology; Testing; Therapeutic; Tobacco; Toxic effect; VDAC1 gene; Work; absorption; acute toxicity; analog; antimicrobial; apoptosis in lymphocytes; base; beta-Lactamase; carbapenem resistance; carbapenem-resistant Enterobacteriaceae; carbapenemase; chemotherapy; comparative; cost; cytokine; design; efflux pump; experience; in vivo; innovation; lead series; microbicide; multi-drug resistant pathogen; nonhuman primate; novel; novel therapeutics; pathogen; pathogenic bacteria; peptide drug; pre-clinical; prototype; resistance factors; resistance mechanism; response; retinal rods; septic; success

The emergence of carbapenem-resistant Enterobacteriaceae (CRE) has created a major and urgent public health threat, because carbapenems are drugs of last resort for treatment of infections caused by a growing number of multidrug resistant (MDR) bacterial pathogens. The objective of this project is to develop a novel therapeutic for treatment of CRE infections by capitalizing on the unique host defense-modulating properties of macrocyclic theta (θ)-defensins in animal models of infection. Preliminary studies provide evidence that θ- defensins and engineered analogs can be developed as agents to treat carbapenemase-producing Klebsiella pneumoniae (KPC-Kp). Pharmacokinetic (PK) studies and analyses of cytokine responses in θ-defensin- treated septic mice demonstrate that these novel cyclic peptides augment host responses to bacteria by modu- lating the release of proinflammatory mediators and attenuating NFκB activation, enhancing host defense in a manner independent of the pathogen’s antibiotic resistance profile. In Aim 1, we will identify a lead therapeutic peptide (LTP) from a series of macrocyclic peptides bioinspired by θ-defensins. Criteria for selection of the LTP will include a) efficacy in the mouse KPC-Kp sepsis model, b) peptide solubility and stability, c) lack of acute toxicity, d) ease of synthetic production, e) extent of bacterial burden reduction in vivo, f) pharmacody- namic (PD) response of additional correlative biomarkers, g) dose-dependent efficacy in a second infection model, and h) single-dose PK. Since half of the lead series of peptides is already in hand, down selection to the LTP can be completed rapidly and certainly within the mandated two-year period. In Aim 2, we will perform single and multiple-dose PK of injectable LTP in mice and NZW rabbits, and use 14C-labeled LTP to define peptide absorption, distribution, metabolism, and excretion (ADME) in mice. Preliminary toxicity studies will be performed to determine maximum tolerated dose in mice. In Aim 3, we propose to express a precyclic-LTP in E. coli using the pET-28a system; the precyclic product is efficiently cyclized in vitro with the recently discov- ered cycloconvertase that produces θ-defensins in vivo. We will also produce cyclic LTP using a robust re- combinant expression system in planta, in collaboration with one of the project’s commercial partners. These studies will capitalize on recent success in producing fully cyclized θ-defensin in tobacco leaf. Collectively, the results of Aim 3 studies will provide a system for efficient, low cost production of the LTP. The significance of the project lies in its promise for development of a unique peptide-based therapeutic for treatment of CRE in- fections. We hypothesize that peptide-based immunomodulatory therapeutics will offer advantages for CRE treatment over agents that seek to target a particular resistance component or function solely as microbicides, representing an innovative approach. Based on published work, existing preliminary data, resources already in place, and established corporate partnerships, we believe the proposed research plan has great potential to introduce a paradigm changing approach for treating CRE and other MDR pathogens.

耐碳青霉烯的肠杆菌科（CRE）的出现创造了一个主要而紧急的公众 健康威胁，因为碳青霉烯是因增长而引起的感染治疗的最后手段的药物 多药耐药（MDR）细菌病原体的数量。该项目的目的是开发小说 通过利用独特的宿主防御调节特性来治疗CRE感染的治疗 在感染动物模型中，大环（θ） - 防御素。初步研究提供了θ-的证据 防御素和工程类似物可以作为治疗产生碳青霉酶的klebsiella的代理 肺炎（KPC-KP）。药代动力学（PK）研究和分析θ-防御素的细胞因子反应 治疗的化粪池小鼠表明，这些新型的环状胡椒体增强了宿主对细菌的反应。 将促炎性介质的释放和衰减NFκB激活剥夺，从而增强了宿主防御 与病原体的抗生素抗性概况无关。在AIM 1中，我们将确定铅疗法 来自θ-防御素的一系列大环生物启动的肽（LTP）。选择的标准 LTP将包括a）小鼠KPC-KP败血症模型的效率，b）肽溶解度和稳定性，c）缺乏 急性毒性，d）易于合成产生，e）体内细菌伯嫩减少的程度，f）药物 - 其他相关生物标志物的NAMIC（PD）响应，G）第二种感染的剂量依赖性效率 模型和H）单剂量PK。由于一半的肽系列已经在手中，因此选择 LTP可以迅速完成，肯定可以在规定的两年内完成。在AIM 2中，我们将表演 小鼠和NZW兔子中注射LTP的单剂量和多剂量PK，并使用14C标记的LTP定义 小鼠的肽滥用，分布，代谢和排泄（ADME）。初步毒性研究将是 执行以确定小鼠中最大耐受剂量。在AIM 3中，我们建议在 使用PET-28A系统的大肠杆菌；预先使用的产品在近期二元期有效地在体外周期性地环绕 在体内产生θ-防御素的ERED环旋转道酶。我们还将使用强大的重新产生环状LTP 与该项目的商业合作伙伴之一合作，Planta中的组合表达系统。这些 研究将利用最近在烟叶中产生完全环化θ-防御素的成功。集体， AIM 3研究的结果将为LTP的有效，低成本生产提供系统。的意义 该项目在于其开发基于肽的独特治疗方法的承诺，用于治疗Cre In- 功能。我们假设基于肽的免疫调节疗法将为CRE提供优势 试图靶向特定抗性成分或仅作为微生物的药物的药物的治疗， 代表一种创新的方法。根据已发表的工作，现有的初步数据，资源已经在 地点并建立了公司合作伙伴关系，我们认为拟议的研究计划具有很大的潜力 引入了一种改变CRE和其他MDR病原体的范式变化方法。