Experimental Animal Models of TB: Chemotherapeutics and Imaging

结核病实验动物模型：化疗和影像学

• 批准号: 10927759
• 负责人: Clifton Barry
• 金额: $ 164.26万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10927759
• 关键词: Adenosine Triphosphate; Advanced Development; Animal Model; Animals; Antibiotics; Bacteria; Benchmarking; Callithrix; Cell Wall; Clinic; Clinical Trials; Data; Development; Disease; Disease Progression; Dose; Drug Combinations; Drug Kinetics; Enzymes; Eukaryotic Cell; Exhibits; Experimental Animal Model; Foundations; Functional Imaging; Granuloma; Human; Image; Imaging Techniques; Individual; Infection; Infection Control; Inflammation; Kinetics; Knowledge; Label; Lesion; Lung; Measures; Metabolism; Modeling; Modernization; Monitor; Mus; Mycobacterium tuberculosis; Mycobacterium tuberculosis antigens; Nitroimidazoles; Organ; Organ Culture Techniques; Oryctolagus cuniculus; Outcome; Pathology; Patients; Penetration; Pharmaceutical Preparations; Positron-Emission Tomography; Pre-Clinical Model; Production; Pyrazinamide; Regimen; Ribosomes; Spatial Distribution; Techniques; Technology; Testing; Therapeutic; Time; Translation Initiation; Translational Repression; Treatment Protocols; Trehalose; Tuberculosis; Work; X-Ray Computed Tomography; bacterial metabolism; bactericide; black hole; chemotherapy; drug candidate; drug distribution; drug efficacy; drug testing; experience; fluorodeoxyglucose; human data; improved; in vivo; inflammatory marker; inhibitor; mutant; neutrophil; nonhuman primate; novel therapeutics; predictive modeling; quinoline; radiotracer; rational design; response; scaffold; secondary metabolite; small molecule; structural imaging; treatment effect; treatment response; tuberculosis chemotherapy; tuberculosis drugs; tuberculosis treatment

This project encompasses approaches to understand how current anti-tubercular chemotherapy works using the most modern technologies and to develop new and improved therapies and therapeutic approaches. Individual projects within this framework are (1) developing structural and functional imaging techniques using PET/CT for use in live, M. tuberculosis (Mtb) infected animals, (2) development of advanced animal models for predicting drug efficacy under conditions that exactly mimic those experienced by TB patients, (3) understanding the activity of various drugs in animal models of tuberculosis therapy, (4) correlating responses seen in animal models with the pathology and response to therapy observed in human TB, and (5) developing techniques for assessing drug distribution, penetration, and pharmacokinetics in vivo. In 2022 and continuing into 2023 we used mice to examine MTB lesions and the function of their neutrophils in relationship to the activity of the anti-TB drug pyrazinamide. In addition, we continued to study the effect of disabling the production of certain secondary metabolites in MTB mutants and found possible effects in host infection control and survival. Most of our PET/CT studies have used 18F-2-fluoro-2-deoxyglucose (FDG) to image the metabolism of the eukaryotic cells in TB lesions in our animal models of tuberculosis. Yet, we would prefer a small molecule that could be used to label Mtb in vivo endogenously as a PET radiotracer. For several years we have focused employing Mtb antigen 85 enzymes that are expressed on the exterior of Mtb cell walls to incorporate exogenous-provided 18F 2-deoxy trehalose (FDT) as either the mono- or dimycolate in or near the cell wall. Use of FDT in the imaging of Mtb in non-human primates, allows the specific imaging of TB-associated lesions and to monitor the effects of treatment in marmosets. In 2023, we and partners began producing the enzymes necessary to finalize FDT characterization including the pharmacokinetics of the probe in NHPs needed to move it toward the clinic. In the past we explored if the marmoset model accurately reflects the response to treatment by providing standard TB treatment HRZE (RIF, INH, PZA, and EMB) to infected symptomatic marmosets and demonstrated that marmosets show similar treatment results as humans. As with the HRZE regimen, during 2022 and finishing up in 2023 we have investigated the combination DBO of Delamanid (D), BDQ (B), and an DprE2 inhibitor OPC-167832 (O) in marmosets in single, double and the triple drug combination. Each individual agent was highly effective promoting survival, reducing pathology, inflammation, and bacterial burdens in the lung and extrapulmonary organs. The 3-drug combination DBO is significantly better than the individual agents and two drug combinations in reducing bacterial burden and organ culture positivity rates. These data have been provided to the stakeholders to share with the FDA. The analysis of the PET/CT derived pathology measures are ongoing, at present, no difference in these measures distinguish DBO from HRZE. We continued to use the mouse, marmoset and rabbit infection models to evaluate classes of antibiotics from our partners in the Gates Foundation's TB Drug Accelerator and other academic partners including diarylquinolines, quinolines, imidazopyridines, nitroimidazoles, among others. These classes of antibiotics are being explored as composing new regimens for treatment of MTB and understanding the specific contribution of each one to activity including consideration of spatial distribution and the kinetics of accumulation in lesions to avoid temporal and spatial black holes of monotherapy. With each new drug candidate, we test for suitable pharmacokinetics, tolerability with longer term dosing (2 weeks to 2 months), in vivo efficacy, and also the candidates penetration into granulomas and cavities to correlate the information with any observed efficacy. We found that the myxobacterial antibiotic myxovalargin was not tolerable in MTB-infected mice although the compound was highly active against MTB, suggesting that while inhibiting translation initiation by occluding the ribosome exit was a viable target, the compound scaffold needed improvement (PMID 36603206). Also in 2023 we specifically examined Bedaquiline (BDQ) and 2 newer diarylquinolines TBAJ876 and TBAJ587, that target the MTB adenosine triphosphate (ATP) synthase enzyme in MTB infected rabbits and found that the new drug candidates had more rapid penetration and faster wash-out periods than BDQ and that one achieved high enough penetration into the center of larger (3-5 cm) lesions to exceed the concentration necessary to kill at least 90% of resident MTB. Both exhibited superior bactericidal activity against caseum resident bacteria than BDQ and were more active in regimens as substitutes for BDQ (PMID 37017524).

该项目包括了解当前抗结核化疗如何使用最现代的技术发挥作用以及开发新的和改进的疗法和治疗方法的方法。该框架内的各个项目包括 (1) 使用 PET/CT 开发结构和功能成像技术，用于活体结核分枝杆菌 (Mtb) 感染动物，(2) 开发先进的动物模型，用于在完全模拟的条件下预测药物疗效结核病患者所经历的那些，（3）了解各种药物在结核病治疗动物模型中的活性，（4）将动物模型中观察到的反应与人类结核病中观察到的病理和治疗反应相关联，以及（5）开发用于结核病治疗的技术评估体内药物分布、渗透和药代动力学。 从 2022 年到 2023 年，我们使用小鼠来检查 MTB 病变及其中性粒细胞的功能与抗结核药物吡嗪酰胺的活性之间的关系。此外，我们继续研究了 MTB 突变体中某些次级代谢产物的丧失产生的影响，并发现了对宿主感染控制和生存的可能影响。 我们的大多数 PET/CT 研究均使用 18F-2-氟-2-脱氧葡萄糖 (FDG) 对结核病动物模型中结核病灶中的真核细胞的代谢进行成像。然而，我们更喜欢一种可以作为 PET 放射性示踪剂在体内内源性标记 Mtb 的小分子。多年来，我们一直致力于使用在 Mtb 细胞壁外部表达的 Mtb 抗原 85 酶，将外源提供的 18F 2-脱氧海藻糖 (FDT) 作为单霉菌酸盐或二霉菌酸盐掺入细胞壁内或细胞壁附近。在非人类灵长类动物中使用 FDT 进行结核分枝杆菌成像，可以对结核病相关病变进行特异性成像，并监测狨猴的治疗效果。 2023 年，我们和合作伙伴开始生产最终确定 FDT 表征所需的酶，包括将其推向临床所需的 NHP 中探针的药代动力学。 过去，我们通过向受感染的症状狨猴提供标准结核病治疗 HRZE（RIF、INH、PZA 和 EMB）来探索狨猴模型是否准确反映了对治疗的反应，并证明狨猴表现出与人类相似的治疗结果。与 HRZE 方案一样，从 2022 年到 2023 年，我们研究了 Delamanid (D)、BDQ (B) 和 DprE2 抑制剂 OPC-167832 (O) 的组合 DBO 在狨猴中的单药、双药和三药组合组合。每种药物都非常有效地促进生存，减少肺部和肺外器官的病理、炎症和细菌负担。 3种药物组合DBO在降低细菌负荷和器官培养阳性率方面明显优于单独药物和两种药物组合。这些数据已提供给利益相关者，以便与 FDA 共享。对 PET/CT 衍生病理学测量的分析正在进行中，目前，这些测量中 DBO 与 HRZE 没有区别。 我们继续使用小鼠、狨猴和兔子感染模型来评估盖茨基金会结核病药物加速器合作伙伴和其他学术合作伙伴提供的抗生素类别，包括二芳基喹啉、喹啉、咪唑并吡啶、硝基咪唑等。正在探索这些类别的抗生素，以制定治疗 MTB 的新方案，并了解每种抗生素对活性的具体贡献，包括考虑空间分布和病灶中积累的动力学，以避免单一疗法的时空黑洞。对于每种新候选药物，我们都会测试其合适的药代动力学、长期给药（2周至2个月）的耐受性、体内功效以及候选药物对肉芽肿和空腔的渗透性，以将信息与任何观察到的功效相关联。我们发现，粘细菌抗生素 myxovalargin 在 MTB 感染的小鼠中无法耐受，尽管该化合物对 MTB 具有高度活性，这表明虽然通过封闭核糖体出口来抑制翻译起始是一个可行的目标，但化合物支架需要改进 (PMID 36603206)。 同样在 2023 年，我们专门检查了贝达喹啉 (BDQ) 和 2 种更新的二芳基喹啉 TBAJ876 和 TBAJ587，它们针对 MTB 感染兔子中的 MTB 腺苷三磷酸 (ATP) 合酶，发现新候选药物具有更快的渗透速度和更快的清除期比 BDQ 更高，并且对较大（3-5 厘米）病灶中心的穿透力足以超过杀死至少所需的浓度90% 的居民 MTB。与 BDQ 相比，两者都对酪乳常驻细菌表现出更优异的杀菌活性，并且在作为 BDQ 替代品的治疗方案中更有效 (PMID 37017524)。

项目成果

Pharmacokinetic evaluation of the penetration of antituberculosis agents in rabbit pulmonary lesions.

抗结核药物在兔肺部病变中渗透的药代动力学评价。

• 发表时间: 2012-01
• 影响因子: 4.9
• 作者: Kjellsson, Maria C;Via, Laura E;Goh, Anne;Weiner, Danielle;Low, Kang Min;Kern, Steven;Pillai, Goonaseelan;Barry 3rd, Clifton E;Dartois, Veronique
• 通讯作者: Dartois, Veronique

Computed Tomography-Based Biomarker for Longitudinal Assessment of Disease Burden in Pulmonary Tuberculosis.

基于计算机断层扫描的生物标志物，用于肺结核疾病负担的纵向评估。

• 发表时间: 2019
• 影响因子: 3.1
• 作者: Gordaliza, P M;Muñoz;Via, L E;Sharpe, S;Desco, M;Vaquero, J J
• 通讯作者: Vaquero, J J

High-sensitivity MALDI-MRM-MS imaging of moxifloxacin distribution in tuberculosis-infected rabbit lungs and granulomatous lesions.

高灵敏度 MALDI-MRM-MS 成像显示结核感染兔肺和肉芽肿病变中莫西沙星的分布。

• 发表时间: 2011-03-15
• 影响因子: 7.4
• 作者: Prideaux, Brendan;Dartois, Véronique;Staab, Dieter;Weiner, Danielle M;Goh, Anne;Via, Laura E;Barry 3rd, Clifton E;Stoeckli, Markus
• 通讯作者: Stoeckli, Markus

A sterilizing tuberculosis treatment regimen is associated with faster clearance of bacteria in cavitary lesions in marmosets.

结核病灭菌治疗方案与狨猴空洞病变中细菌的更快清除有关。

• 发表时间: 2015-07
• 影响因子: 4.9
• 作者: Via, Laura E;England, Kathleen;Weiner, Danielle M;Schimel, Daniel;Zimmerman, Matthew D;Dayao, Emmanuel;Chen, Ray Y;Dodd, Lori E;Richardson, Mike;Robbins, Katherine K;Cai, Ying;Hammoud, Dima;Herscovitch, Peter;Dartois, Véronique;Flynn, JoAnne
• 通讯作者: Flynn, JoAnne

Meropenem-clavulanic acid shows activity against Mycobacterium tuberculosis in vivo.

美罗培南克拉维酸在体内表现出抗结核分枝杆菌的活性。

• 发表时间: 2012-06
• 影响因子: 4.9
• 作者: England, Kathleen;Boshoff, Helena I M;Arora, Kriti;Weiner, Danielle;Dayao, Emmanuel;Schimel, Daniel;Via, Laura E;Barry 3rd, Clifton E
• 通讯作者: Barry 3rd, Clifton E