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: 9912709
• 负责人: Andre J. Ouellette
• 金额: $ 89.96万
• 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 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9912709
• 关键词: ADME Study; Acute Toxicity Tests; Animal Model; Antibiotic Resistance; Antibiotics; Attenuated; Bacteria; Bacterial Infections; Binding; Biochemical; Biological Markers; Carbapenems; Characteristics; Collaborations; Complement; Complex; Cyclic GMP; Cyclic Peptides; Development; Dose; Drug Kinetics; Engineering; Enterobacteriaceae; Enterobacteriaceae Infections; Epidemic; Escherichia coli; Evaluation; Excretory function; Goals; Hand; Host Defense; Immune; Immune response; Immune system; Immunity; Immunomodulators; In Vitro; Inbred BALB C Mice; Infection; Injectable; Kilogram; Kinetics; Klebsiella; Klebsiella pneumoniae; Knowledge; Label; Lead; Leukocytes; Lymphocyte Suppression; Maximum Tolerated Dose; Mediating; Mediator of activation protein; Membrane; Metabolism; Methods; Modeling; Multi-Drug Resistance; Multiple Bacterial Drug Resistance; Mus; Oryctolagus cuniculus; Pan Genus; Penicillins; Peptides; Periodicity; Pharmaceutical Preparations; Pharmacodynamics; Pharmacologic Substance; Plant Leaves; Plants; Production; Property; Public Health; Publishing; Rattus; Recombinants; Reporting; Research; Resistance; Resistance profile; Resort; Rod; Selection Criteria; Sepsis; Series; Solubility; Solvents; System; Technology; Testing; Therapeutic; Therapeutic Uses; Tobacco; Toxic effect; VDAC1 gene; Work; absorption; acute toxicity; alpha-Defensins; analog; antimicrobial; apoptosis in lymphocytes; base; beta-Lactamase; carbapenem resistance; carbapenem-resistant Enterobacteriaceae; carbapenemase; chemotherapy; comparative; cost; cytokine; data resource; design; efflux pump; experience; immunoregulation; in vivo; innovation; lead series; microbicide; multi-drug resistant pathogen; nonhuman primate; novel; novel therapeutics; overtreatment; pathogen; pathogenic bacteria; peptide drug; pharmacokinetics and pharmacodynamics; pre-clinical; preclinical development; prototype; resistance mechanism; response; septic; success; theta-defensin

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.

碳青霉烯类耐药肠杆菌科（CRE）的出现，已造成重大和紧迫的公众 健康威胁，因为碳青霉烯类是治疗日益严重的 多药耐药（MDR）细菌病原体的数量。这个项目的目标是开发一个新的 通过利用CRE的独特宿主防御调节特性， 大环θ-防御素在动物感染模型中的作用。初步研究表明，θ- 防御素和工程化类似物可被开发为治疗产生碳青霉烯酶的克雷伯氏菌的药剂 肺炎克雷伯氏菌（KPC-Kp）。药代动力学（PK）研究和θ-防御素中细胞因子应答的分析- 治疗的脓毒症小鼠证明，这些新的环肽通过modu- 抑制促炎介质的释放，减弱NFκB的激活，增强宿主的防御能力， 方式独立于病原体的抗生素耐药性谱。在目标1中，我们将确定一种领先的治疗方法， 肽（LTP）是从一系列受θ-防御素生物启发的大环肽中合成的。甄选标准 LTP将包括a）在小鼠KPC-Kp脓毒症模型中的功效，B）肽溶解度和稳定性，c）缺乏免疫原性。 急性毒性，d）合成生产的容易性，e）体内细菌负荷减少的程度，f）药理学， g）在第二次感染中的剂量依赖性功效 模型，和h）单次给药PK。由于一半的领先系列肽已经在手，向下选择， LTP可以迅速完成，并肯定在规定的两年期限内完成。在目标2中，我们将执行 在小鼠和NZW兔中注射LTP单次和多次给药PK，并使用14 C标记的LTP来定义 肽在小鼠中的吸收、分布、代谢和排泄（ADME）。初步毒性研究将在 以确定小鼠中的最大耐受剂量。在目标3中，我们提出用以下方式表达前循环LTP： E.大肠杆菌中使用pET-28 a系统;预环产物在体外有效地环化与最近发现的， 在体内产生θ-防御素的环状转化酶。我们还将使用一个强大的重新生成循环LTP， 植物中的组合表达系统，与该项目的商业合作伙伴之一合作。这些 研究将利用最近在烟草叶中产生完全环化的θ-防御素的成功。统称 目标3研究的结果将提供一种有效、低成本生产LTP的系统。的意义 该项目有望开发一种独特的基于肽的治疗方法，用于治疗CRE， 感染。我们假设基于肽的免疫调节疗法将为CRE提供优势 针对特定耐药性成分或仅作为杀微生物剂的药物治疗， 代表了一种创新的方法。根据已发表的工作、现有的初步数据、 地方，并建立了企业合作伙伴关系，我们相信拟议的研究计划有很大的潜力， 介绍了一种治疗CRE和其他MDR病原体的范式改变方法。