Targeting Biotin Metabolism in Mycobacterium Tuberculosis

靶向结核分枝杆菌中的生物素代谢

基本信息

批准号：
10322125
负责人：
Courtney C Aldrich
金额：
$ 77.99万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-01-15 至 2023-12-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10322125
关键词：
AIDS-Related Opportunistic Infections Acute Anabolism Antitubercular Agents Atypical Mycobacteria Bacillus Biochemical Biological Assay Biological Availability Biotin Biotin Metabolism Pathway Cells Cellular Structures Chronic Communicable Diseases Data Drug Interactions Drug Kinetics Drug Targeting Drug resistance Ensure Enzyme Inhibition Enzymes Etiology Evaluation Extreme drug resistant tuberculosis Fluorine Generations Genetic Goals Knowledge Lead Ligase Ligation Measures Metabolism Microbiology Minnesota Molecular Conformation Multi-Drug Resistance Multiple drug resistant Mycobacteria Tuberculosis Mus Mycobacterium tuberculosis Natural Products Oral Pathway interactions Patients Pharmaceutical Chemistry Pharmaceutical Preparations Property Proteins Regimen Research Resistance Safety Structure Tuberculosis Universities Validation Work absorption analog base cofactor combat design drug discovery drug disposition efficacy study extensive drug resistance genetic approach genome sequencing global health improved in vivo inhibitor medical schools mortality mouse model mutant mycobacterial nanomolar novel therapeutics overexpression pathogen pre-clinical preclinical development programs resistance frequency resistance mechanism resistant strain small molecular inhibitor synergism treatment duration tuberculosis drugs whole genome

项目摘要

SUMMARY Mycobacterium tuberculosis (Mtb), the principal etiological agent of tuberculosis (TB), infects over one-third of humanity and is now the leading cause of infectious disease mortality by a single pathogen. Mtb requires biotin for survival and synthesizes this essential cofactor de novo. In preliminary studies using a genetic approach, we have shown biotin biosynthesis and ligation are essential for Mtb infection in mice. We have synthesized a selective nanomolar inhibitor of biotin protein ligase termed Bio-AMS that targets the enzyme biotin protein ligase (BPL,) responsible for the ligation of biotin onto biotin-dependent enzymes. We have also identified the natural product acidomycin, which targets the final step of biotin biosynthesis catalyze by BioB. However, Bio- AMS and acidomycin have liabilities in their drug disposition properties leading to rapid clearance, poor volume of distribution, and limited oral bioavailability. There are also gaps in our knowledge regarding their mechanism of resistance and activity when combined with other TB drugs. The objectives of this application are: 1) to develop our lead compounds Bio-AMS and acidomycin through the optimization of their ADME (absorption, distribution, metabolism and elimination) properties and pharmacokinetic parameters into viable preclinical candidates, 2) to more deeply illuminate the mechanism of action and resistance in Mtb, 3) to determine the safety profile and potential drug-drug interactions, and 4) to identify interactive effects with other TB drugs (i.e. synergy). We will accomplish the overall objectives of this application by pursuing three specific aims. In aim 1, we will carry out an iterative structure-based medicinal chemistry program of Bio-AMS and acidomycin to concurrently optimize pharmacokinetic (PK) parameters and whole-cell activity using a combination of approaches including fluorination, structural simplification, and introduction of conformation constraints. In aim 2, we will perform biochemical and cellular studies to evaluate enzyme inhibition, target engagement, cellular accumulation, and whole-cell activity against Mtb as well as drug-resistant strains. Generation of resistant strains followed by whole-genome sequencing will be used to characterize potential resistance mechanisms and determine the resistance frequency. Finally, combination studies with various first and second-line TB drugs will be undertaken to assess potential for synergy. In aim 3, the Bio-AMS and acidomycin analogues will be assessed in vivo to determine their complete pharmacokinetic parameters with a goal to improve on the volume of distribution (Vd), intrinsic clearance (CL), and bioavailability (F). We will evaluate compounds against a panel of assays (hERG, CYP inhibition, Ames mutagenicity) to ensure safety and selectivity. In vivo efficacy studies will be done using murine models of acute and chronic TB infection

概括结核分枝杆菌（MTB），结核病的主要病因（TB），感染了超过三分之一的三分之一人类，现在是单个病原体传染病死亡率的主要原因。 MTB需要生物素为了生存并综合了这个必不可少的辅助因子。在使用遗传学方法的初步研究中，我们已经表明，生物素的生物合成和结扎对于小鼠的MTB感染至关重要。我们已经合成了生物素蛋白连接酶的选择性纳摩尔抑制剂称为生物AM，靶向酶生物素蛋白连接酶（BPL，）负责生物素连接到生物素依赖性酶。我们还确定了天然产物酸霉素，它针对生物素生物合成的最后一步催化了Biob。但是，生物 - AMS和酸霉素在其药物处置特性中具有责任，导致清除率较差，体积较差分销和有限的口服生物利用度。我们关于其机制的知识中也存在差距与其他结核病药物结合使用时的耐药性和活性。此应用程序的目标是：1）开发我们的铅化合物通过优化其ADME（吸收，，分布，代谢和消除）特性和药代动力学参数到可行的临床前候选人，2）更深入地阐明MTB中的作用和抵抗机理，3）确定安全性和潜在的药物相互作用，以及4）确定与其他结核病药物的互动效应（即协同作用）。我们将通过追求三个特定目标来实现此应用程序的总体目标。在AIM 1中，我们将执行基于迭代结构的生物AM和酸霉素的药物化学计划同时优化药代动力学（PK）参数和全细胞活性，结合包括荧光，结构简化和构象约束的引入的方法。目标 2，我们将进行生化和细胞研究以评估酶抑制，靶标，细胞积累和针对MTB的全细胞活性以及抗药性菌株。产生抗性菌株随后进行全基因组测序将用于表征潜在的电阻机制并确定电阻频率。最后，与各种一线和二线结核的组合研究将进行药物以评估协同作用的潜力。在AIM 3中，生物AM和酸霉素类似物将在体内评估以确定其完整的药代动力学参数，以改进分布量（VD），内在清除率（CL）和生物利用度（F）。我们将评估化合物针对一系列测定面板（HERG，CYP抑制，AMES诱变），以确保安全性和选择性。体内疗效研究将使用急性和慢性结核病感染的鼠模型进行