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.

耐碳青霉烯类肠杆菌科细菌(CRE)的出现已造成重大而紧迫的公众 威胁健康，因为碳青霉烯类是治疗由日益增长的 多药耐药(MDR)细菌病原体的数量。这个项目的目标是开发一种新的 利用Cre独特的宿主防御调节特性治疗Cre感染 大环角蛋白(θ)--感染动物模型中的防御素。初步研究提供证据表明θ- 防御素和工程类似物可以被开发为治疗产生碳青霉烯酶的克雷伯菌的药物 肺炎(KPC-KP)。θ-防御素的药代动力学研究及细胞因子反应分析 经治疗的脓毒症小鼠表明，这些新的环肽通过调节宿主对细菌的免疫应答而增强宿主对细菌的反应。 延缓炎性介质释放和抑制核因子κB的激活，增强宿主防御 与病原体的抗生素耐药性特征无关的方式。在目标1中，我们将确定一种铅治疗方法 受θ-防御素生物启发的一系列大环肽。评选准则 LTP将包括a)在小鼠KPC-KP脓毒症模型中的疗效，b)多肽的溶解性和稳定性，c)缺乏 急性毒性，d)合成生产的简易性，e)体内细菌负荷减少的程度，f)药理- 附加相关生物标志物的动态(PD)反应，g)二次感染的剂量依赖效应 模型，h)单剂PK。由于先导系列多肽的一半已经在手中，向下选择到 长期合作伙伴关系可以迅速完成，当然也可以在规定的两年期限内完成。在《目标2》中，我们将表演 小鼠和新西兰兔注射LTP的单剂量和多剂量PK，并用~(14)C标记的LTP确定 小鼠体内多肽吸收、分布、代谢和排泄(ADME)。初步毒性研究将是 以确定小鼠的最大耐受量。在目标3中，我们建议将预循环-LTP表示为 利用pET-28a系统构建的pET-28a系统在体外能有效地环化前环产物。 在体内产生θ防御素的环状转换酶。我们还将使用健壮的Re-Re来生成循环LTP Planta中的组合表达系统，与该项目的一个商业合作伙伴合作。这些 研究将利用最近在烟叶中生产完全环化的θ-防御素的成功。总体而言， AIM 3的研究结果将为高效、低成本生产LTP提供一个系统。的重要意义。 该项目承诺开发一种独特的基于多肽的治疗方法，用于治疗慢性粒细胞白血病。 美味佳肴。我们假设，基于多肽的免疫调节疗法将为CRE提供优势 对寻求靶向特定抗药性成分或仅起杀微生物剂作用的药物的治疗， 代表了一种创新的方法。基于已发表的工作，现有的初步数据，已经在 ，并建立了公司伙伴关系，我们相信拟议的研究计划具有巨大的潜力 介绍一种治疗CRE和其他MDR病原体的范式改变方法。