Biomimetic Peptide Aerosols for Rapid Clearance of Pulmonary MDR Tuberculosis

项目总结 耐多药结核病(MDR-TB)，约占复发结核病病例的20%，被诊断为 每年有40万名患者，这是一个紧迫的全球卫生优先事项，有可能破坏美国的结核病 淘汰策略。耐多药结核病传播的关键是干扰和不遵守标准治疗 疗程长(长达24个月)，每天需要大剂量的抗生素。这个项目的目标是 是开发一种可雾化的窄谱抗菌生物材料，可以与批准的结核病配对 抗生素可迅速清除肺部耐多药结核病，并大幅缩短疗程。基本原理 对于这一策略，是一种新的类蛋白质模拟宿主防御肽，我们已经设计了从头开始经历 在结核分枝杆菌(Mtb)富含霉菌酸的外膜内指示自组装。我们 已经表明，我们的主要候选者MAD1在暴露几分钟内就会引起结核病特异性的细菌溶解，而不是 对保护性呼吸道共生区和宿主肺组织的附带毒性。此外，这些新奇的多肽 协同增强临床抗生素的活性，实现纳米分子抗结核效果。然而，这些 合成肽具有药代动力学责任，包括快速清除和有限的肺 生物利用度，我们对它们结合时的作用机制的了解仍然存在空白 与其他药物有关。本申请的目的是：(I)更深入地调查行动模式(MOA) 以及我们的先导化合物MAD1在Mtb中的药物相互作用(例如，协同作用)，(Ii)改善其ADME(吸收， 分布、代谢和消除)性质和药代动力学参数 优化和配制成新的生物材料气雾剂，以及(Iii)确定 与疾病相关的临床前模型中的铅配方。我们将分三个目标实现这些目标。 在目标1中，人工智能指导的基于结构的序列筛选和重组工程分析将 优化MAD1‘S对结核分枝杆菌和耐药菌株的效力，并通报MOA。全基因组 在这些研究中产生的耐药菌株的测序将表征可能的耐药机制 并确定电阻频率。目标2将开发MAD1和抗生素的可吸入制剂 利用我们专有的气凝胶输送系统，旨在利用结核分枝杆菌的关键代谢脆弱性 和病原体特异性肺部治疗。联合细菌学研究将评估协同作用的潜力 巨噬细胞对耐多药结核和多药耐药细胞的杀伤作用。在目标3中，我们评估了肺部的药代动力学 以优化肺生物利用度为目标的优先气凝胶配方的参数和 治疗载体的滞留/清除动力学。我们将评估治疗制剂的安全性 通过一系列分析(组织学、肺功能、免疫原性)并评估几种 小鼠急性和慢性结核病感染模型。

PROJECT SUMMARY Multidrug-resistant Tuberculosis (MDR-TB), which accounts for ~20% of recurrent TB cases and is diagnosed in 400,000 patients each year, represents an urgent global health priority that threatens to undermine US TB elimination strategies. Key to MDR-TB transmission is disruption and non-compliance with standard therapeutic regimens, which are lengthy (up to 24 months) and require high daily doses of antibiotics. The goal of this project is to develop an aerosolizable, narrow-spectrum antimicrobial biomaterial that can be paired with approved TB antibiotics to rapidly clear pulmonary MDR-TB and dramatically shorten the course of treatment. Fundamental to this strategy is a new class of protein-mimetic host defense peptides we have engineered de novo to undergo instructed self-assembly within the mycolic-acid rich outer membrane of Mycobacterium tuberculosis (Mtb). We have shown that our lead candidate, MAD1, elicits TB-specific bacteriolysis within minutes of exposure, without collateral toxicity towards protective respiratory commensals and host lung tissue. Further, these novel peptides synergistically enhance the activity of clinical antibiotics to achieve nanomolar anti-TB efficacy. However, these synthetic peptides have pharmacokinetic liabilities that include rapid clearance and limited pulmonary bioavailability, and there remain gaps in our knowledge regarding their mechanism of action when combined with other drugs. The objectives of this application are to: (i) more deeply investigate the mode of action (MoA) and drug interactions (e.g. synergy) of our lead compound MAD1 in Mtb, (ii) improve its ADME (absorption, distribution, metabolism and elimination) properties and pharmacokinetic parameters through sequence optimization and formulation into novel biomaterial aerosols, and (iii) determine the safety profile and efficacy of lead formulations in disease-relevant preclinical models. We will accomplish these objectives over three aims. In aim 1, artificial intelligence-guided structure-based sequence screening and recombineering assays will optimize MAD1’s potency against Mtb and drug-resistant strains, as well as inform on MoA. Whole-genome sequencing of resistant strains generated during these studies will characterize possible resistance mechanisms and determine the resistance frequency. Aim 2 will develop inhalable formulations of MAD1 and antibiotics utilizing our proprietary aerogel delivery system designed to exploit a key metabolic vulnerability of Mtb for rapid and pathogen-specific pulmonary therapy. Combination bacteriologic studies will assess potential for synergy towards MDR-TB and persister cells in macrophages. In aim 3, we assess the pulmonary pharmacokinetic parameters of prioritized aerogel formulations with the goal of optimizing the lung bioavailability and residence/clearance kinetics of the therapeutic carrier. We will evaluate the safety of therapeutic formulations via a series of assays (histology, pulmonary function, immunogenicity) and assess in vivo efficacy in several murine models of acute and chronic TB infection.