Optimizing Multi-drug Mycobacterium tuberculosis Therapy for Rapid Sterilization and Resistance Suppression

优化结核分枝杆菌多药治疗以实现快速灭菌和耐药性抑制

• 批准号: 10567327
• 负责人: George Louis Drusano
• 金额: $ 131.43万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-25 至 2027-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10567327
• 关键词: Address; Algorithms; Amino Acyl-tRNA Synthetases; Animals; Antimicrobial Effect; Bacteria; Binding; Biological Assay; C3HeB/FeJ Mouse; Carbapenems; Catabolism; Cell Wall; Cells; Cholesterol; Code; Combination Drug Therapy; Combined Modality Therapy; Complex; Data; Drug Combinations; Drug Exposure; Equation; Evaluation; Excision; Experimental Designs; Fiber; Foundations; Human; Image; In Vitro; Inbred BALB C Mice; Infection; Kinetics; Lesion; Link; Measures; Metabolic; Modeling; Monobactams; Mus; Mycobacterium tuberculosis; New Agents; Oral; Organism; Pathologic; Pathology; Pathway interactions; Patients; Penetration; Pharmaceutical Preparations; Phase; Poison; Population; Predisposition; Primary Infection; Property; Publishing; Regimen; Resistance; Rest; Site; Spatial Distribution; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Speed; Sterilization; Structure of parenchyma of lung; Testing; Time; Tissues; Tuberculosis; Validation; Work; Writing; base; beta-Lactams; cell killing; cohort; data modeling; dosage; drug distribution; efficacy study; experimental study; flasks; high dimensionality; in vivo; inhibitor; insight; laser capture microdissection; leucine-tRNA; liquid chromatography mass spectroscopy; man; mass spectrometric imaging; mathematical methods; mathematical model; mouse model; nonhuman primate; novel; prospective; receptor binding; response; success; supercomputer; therapy duration; tuberculosis treatment

Project Summary/Abstract In P01 AI123036, we were able to generate an algorithm that ranked single agents for Mycobacterium tuberculosis (MTB), identified promising 2-drug combinations and, with a completely novel mathematical approach, identified 3-drug regimens predicted to be significantly better than 2-drug regimens. These predictions were prospectively validated in a BALB/c model (H37Rv) and in a Non-Human Primate model of MTB (Erdman strain). In this proposal, we will extend our previous work. There is a large number of new MTB agents, many with novel mechanisms of action. We have 4 Specific Aims (SA) that, when complete, will allow us to identify multi-drug combinations that will optimize rate of kill for organisms in 3 different metabolic states and will suppress resistance emergence. In the Hollow Fiber Infection Model [HFIM] (SA#1), we will be able to rank new agents on the bases of potency and physicochemical properties. The HFIM provides insight into the drug’s exposure-response for kill and resistance suppression. We identified a near optimal 3-drug regimen (PMD/MFX/BDQ). With new single agents, we can examine substituting a new agent for an older agent AND we can expand the regimens to identify a near- optimal 4-drug regimen. This will be particularly important for patients with high bacterial burdens. In SA #2, we will test regimens from SA#1 in two murine models (BALB/c & C3HeB/FeJ mice). These will give somewhat different information. Both give information regarding kill and resistance suppression. Kramnik mice have pathology more closely resembling that in humans. We will use Matrix-Assisted Laser Desorption Ionization-MS Imaging and Laser Capture Microdissection LCMS. This allows identification of spatial distribution and quantification of drugs. A question regarding cure is how long to wait to sacrifice animals to document eradication. Some agents (BDQ) have long tissue half-lives. We will document rates of ingress/egress of drugs into the infection site, allowing determination when animal cohorts may be sacrificed to document eradication. In SA #3, we will document mechanisms of antimicrobial effect quantitatively. We have generated a first-of-a- kind dynamic model for PBP-binding in MTB, and will link this to rates of cell kill. We have also developed AMP/ADP/ATP intracellular assays. These will be employed for agents like diarylquinolines (e.g. BDQ) and PMD that act as energy poisons (for PMD, this occurs under anaerobic/non-replicative conditions. We will measure intracellular (MTB) drug concentrations, linking them to effect alone and in combination therapy experiments. Proposal success rests on modeling of the data. In SA #4, we have written code to extend earlier analyses, going from 3- to 4-drug regimens. For these high dimensional models, we developed several approaches to speed up analysis making them computationally tractable. At proposal end, we shall develop a 4-drug algorithm allowing rapid identification of near optimal regimens that work for both susceptible and less-susceptible organisms. The algorithm will be general. It will work well for today’s agents but also for agents as discovered.

项目摘要/摘要 在P01 AI123036中，我们能够生成一种对分枝杆菌进行单剂的算法 结核病（MTB），确定了有希望的2毒素组合，并具有完全新颖的数学 预测的3级药物疗法明显优于2级药物方案。这些预测 可能在BALB/C模型（H37RV）和MTB的非人类灵长类动物模型中进行了验证（Erdman 拉紧）。在此提案中，我们将扩展以前的工作。 有大量新的MTB代理，许多代理具有新颖的作用机理。我们有4个具体目标 （SA）完成后，将使我们能够识别多药组合，以优化杀人率 3个不同代谢状态的生物体将抑制抗药性出现。 在空心纤维感染模型[HFIM]（SA＃1）中，我们将能够在效力基础上对新代理进行排名 和物理特性。 HFIM提供了有关该药物的杀戮和杀戮暴露反应的见解 阻力抑制。我们确定了一种接近最佳的3毒剂治疗方案（PMD/MFX/BDQ）。有了新的单个代理商， 我们可以检查替代新代理商的新代理，我们可以扩展该方案以识别几乎 - 最佳的4级药物方案。这对于伯良细菌较高的患者尤为重要。 在SA＃2中，我们将测试两种鼠模型中的SA＃1（BALB/C＆C3HEB/FEJ小鼠）的方案。这些会给予 有些不同的信息。两者都提供有关杀戮和抑制抑制的信息。 Kramnik小鼠 在人类中，使病理更相似。我们将使用矩阵辅助激光解吸 电离-MS成像和激光捕获显微解剖LCM。这允许识别空间分布 和药物的定量。关于治愈的问题是要等待多长时间牺牲动物来记录 根除。一些试剂（BDQ）具有长组织半衰期。我们将记录药物的入学率/出口率 进入感染部位，允许何时牺牲动物队列以记录消除。 在SA＃3中，我们将定量地记录抗微生物有效的机制。我们已经产生了一个 MTB中PBP结合的类型动态模型，并将其与细胞杀伤率联系起来。我们也发展了 AMP/ADP/ATP细胞内测定。这些将被用于牙龈二元（例如BDQ）和PMD等代理商 该充当能量毒物（对于PMD，这是在厌氧/非复制条件下发生的。我们将测量 细胞内（MTB）药物浓度，将它们与单独作用和联合治疗实验联系起来。 提案成功取决于数据的建模。在SA＃4中，我们已经书面代码来扩展早期分析， 从3-到4级药物方案。对于这些高维模型，我们开发了几种方法 加快分析，使它们在计算上可以处理。在提案结束时，我们将开发4级药物算法 允许快速识别易于易受感染和易感性的近乎最佳方案 有机体。该算法将是一般。它适用于当今的代理商，也适用于发现的代理商。