A Novel Target for New Anti-Anthrax Drugs

新型抗炭疽药物的新靶点

• 批准号: 7169645
• 负责人: WAYNE J BROUILLETTE
• 金额: $ 34.37万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7169645
• 关键词: Anabolism; Anthrax disease; Anti-Bacterial Agents; Antibiotic Resistance; Azides; Bacillus (bacterium); Bacillus anthracis; Bacillus cereus; Bacillus subtilis; Binding Sites; Biological Assay; Biological Warfare; Cell Membrane Permeability; Class; Crystallography; Differential Scanning Calorimetry; Disruption; Dissociation; Drug Delivery Systems; Energy Metabolism; Enzyme Inhibition; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzyme Kinetics; Enzymes; Equilibrium; Erythrocytes; Evaluation; Exhibits; Gel Chromatography; Generations; Goals; Gram-Negative Bacteria; Growth; Human; In Vitro; Infection; Inhibitory Concentration 50; Lead; Libraries; Ligase; Measures; Molecular; Multienzyme Complexes; Nicotinamide adenine dinucleotide; Numbers; Other Biochemical Pathway; Pathway interactions; Pharmaceutical Preparations; Phase; Photoaffinity Labels; Process; Property; Proteins; Range; Research Personnel; Score; Screening procedure; Security; Solutions; Structure; Structure-Activity Relationship; Synthesis Chemistry; Terrorism; Therapeutic Agents; chemical synthesis; cofactor; design; high throughput screening; improved; in vitro Assay; inhibitor/antagonist; monomer; novel; prevent; programs; protein protein interaction; research study; small molecule; transport inhibitor; virtual

DESCRIPTION (provided by applicant): The use of Bacillus anthracis in acts of terrorism and/or biological warfare is a demonstrated threat to U.S. security. The long-term objective of this proposal is to develop inhibitors against a new drug target, the anthracis enzyme nicotinamide adenine dinucleotide (NAD) synthetase, that are effective therapeutic agents for preventing and/or treating infections caused by B. anthracis. NAD synthetase catalyzes the last step in both the de novo and salvage pathways for the biosynthesis of NAD, an essential cofactor in energy metabolism. Since exogenous NAD cannot support bacterial growth due to insufficient cell membrane permeability/transport, inhibitors of NAD biosynthesis should be bacteriostatic and/or bacteriocidal. We have identified the first low micromolar inhibitors of Bacillus NAD synthetase, and these effectively inhibit the vegetative growth of Bacillus anthracis, and are bacteriocidal, at concentrations around 1 (mu/g/mL. We will now perform reiterative design, chemical synthesis, and in vitro analysis to develop mature lead compounds. The Specific Aims are: (1) Using parallel, solution phase synthetic chemistry, we will optimize inhibitory activities for existing classes of lead structures until low nanomolar inhibitors are obtained. (2) Determine the molecular mechanism by which existing lead synthetic compounds inhibit NAD synthetase. In addition to protein crystallography, we will: (a) Measure the Ki and evaluate the type of inhibition for synthetic inhibitors of NAD synthetase. (b) Characterize the enzyme homodimer/monomer equilibrium and modulation by inhibitors. (c) Perform photoaffinity labeling experiments using inhibitors containing a photoreactive alkylating group (azide). (3) Alternative structural templates will be developed as new NAD synthetase inhibitors. All synthetic compounds will be evaluated in high throughput screens as enzyme inhibitors (IC50 and/or Ki) and antibacterials (MIC). Selectivity for the inhibition of Bacillus NAD synthetase over the human (erythrocyte) enzyme will also be evaluated.

描述（由申请人提供）：在恐怖主义和/或生物战行为中使用炭疽杆菌已证明对美国安全构成威胁。该提案的长期目标是开发针对新药物靶标炭疽酶烟酰胺腺嘌呤二核苷酸（NAD）合成酶的抑制剂，这是预防和/或治疗炭疽芽孢杆菌引起的感染的有效治疗剂。 NAD 合成酶催化 NAD 生物合成的从头途径和补救途径的最后一步，NAD 是能量代谢中的重要辅助因子。由于细胞膜渗透性/运输不足，外源 NAD 不能支持细菌生长，因此 NAD 生物合成抑制剂应该具有抑菌和/或杀菌作用。 我们已经鉴定出第一个低微摩尔的芽孢杆菌NAD合成酶抑制剂，这些抑制剂可以有效抑制炭疽杆菌的营养生长，并且具有杀菌作用，浓度约为1（μ/g/mL）。现在我们将进行反复设计、化学合成和体外分析开发成熟的先导化合物的具体目标是：(1) 使用平行的溶液相合成化学，我们将优化现有类别先导结构的抑制活性，直至降低。 (2) 确定现有先导合成化合物抑制 NAD 合成酶的分子机制 除了蛋白质晶体学之外，我们还将： (a) 测量 Ki 并评估 NAD 合成酶合成抑制剂的抑制类型。 (b) 表征酶同二聚体/单体平衡和抑制剂的调节。 (c) 使用含有光反应性烷基化基团（叠氮化物）的抑制剂进行光亲和标记实验。 (3) 将开发替代结构模板。作为新型NAD合成酶抑制剂。 所有合成化合物都将在高通量筛选中作为酶抑制剂（IC50 和/或 Ki）和抗菌剂（MIC）进行评估。还将评估芽孢杆菌 NAD 合成酶相对于人类（红细胞）酶的抑制选择性。

项目成果

Tethered dimer inhibitors of NAD synthetase: parallel synthesis of an aryl-substituted SAR library.

NAD 合成酶的束缚二聚体抑制剂：芳基取代的 SAR 文库的平行合成。

• DOI: 10.1021/cc050063j
• 发表时间: 2005
• 期刊: Journal of combinatorial chemistry.
• 作者: Velu,SadanandanE;Luan,Chi-Hao;Delucas,LawrenceJ;Brouillette,ChristieG;Brouillette,WayneJ
• 通讯作者: Brouillette,WayneJ

SAR studies for a new class of antibacterial NAD biosynthesis inhibitors.

• DOI: 10.1021/cc9000357
• 发表时间: 2009-07
• 期刊: Journal of combinatorial chemistry
• 作者: Moro WB;Yang Z;Kane TA;Zhou Q;Harville S;Brouillette CG;Brouillette WJ
• 通讯作者: Brouillette WJ