Rational Design of Adenylation Enzyme Inhibitors

说明（由申请人提供）：腺苷酸化酶抑制剂的合理设计腺苷酸化酶在跨越所有三个生命领域的多种生物途径中发挥中心作用，介导蛋白质翻译、脂肪酸和氨基酸代谢、天然产物生物合成和泛素结合等过程。这些酶催化两步反应，涉及羧酸底物的初始ATP依赖性腺苷酸化以形成紧密结合的酰基-AMP（酰基腺苷酸）中间体，随后亲核试剂攻击该混合酸酐以形成酯、硫酯或酰胺产物。引人注目的是，虽然所有的腺苷酸化酶催化相同的一般反应，至少有六个不同的蛋白质折叠已被确定在这个机械超家族。腺苷酸化酶的抑制剂在感染性疾病、癌症、心血管疾病、代谢疾病、免疫病理学和神经退行性疾病中具有重要的潜在生物医学应用。我们在此建议继续我们成功的，长期的计划，合理设计腺苷酸化酶抑制剂。我们正在推进一个抑制剂设计平台，该平台利用机制和结构信息，并适用于所有类别的腺苷酸化酶。我们以前的努力已经产生了针对细菌铁载体，糖脂和甲基萘醌生物合成的新型抗菌药物。我们还开发了泛素/泛素样修饰剂E1激活酶的半合成蛋白质抑制剂，为这些酶的功能提供了深刻的机理见解。我们下一个项目期的目标是使用大环设计开发非核糖体肽生物合成的选择性抑制剂，开发优化的甲基萘醌生物合成抑制剂作为新的抗菌药物，并开发基于蛋白质和小分子的E1活化酶抑制剂，以探索其在癌症中的分子机制和生物学功能。还将研究理化性质与细菌摄取的系统相关性。这项工作将对基于结构的设计、抗菌药物化学和酶学产生广泛影响，并将通过已建立的多学科合作进行，包括有机合成、药物化学、药理学、生物化学、微生物学和结构生物学方面的综合专业知识。

DESCRIPTION (provided by applicant): Rational Design of Adenylation Enzyme Inhibitors Adenylation enzymes play central roles in diverse biological pathways across all three domains of life, mediating processes such as protein translation, fatty acid and amino acid metabolism, natural product biosynthesis, and ubiquitin conjugation. These enzymes catalyze a two-step reaction involving initial ATP- dependent adenylation of a carboxylic acid substrate to form a tightly-bound acyl-AMP (acyl adenylate) intermediate, followed by attack of a nucleophile on this mixed anhydride to form an ester, thioester, or amide product. Strikingly, while all adenylation enzymes catalyze this same general reaction, at least six distinct protein folds have been identified within this mechanistic superfamily. Inhibitors of adenylation enzymes have important potential biomedical applications in infectious diseases, cancer, cardiovascular disease, metabolic disease, immunopathologies, and neurodegenerative disorders. We propose herein to continue our successful, long-term program on the rational design of adenylation enzyme inhibitors. We are advancing an inhibitor design platform that leverages mechanistic and structural information and is general for all classes of adenylation enzymes. Our previous efforts have yielded novel antibacterials targeting bacterial siderophore, glycolipid, and menaquinone biosynthesis. We have also developed semisynthetic protein inhibitors of ubiquitin/ubiquitin-like modifier E1 activating enzymes that have provided profound mechanistic insights into the functions of these enzymes. Our goals for the next project period are to develop selective inhibitors of non-ribosomal peptide biosynthesis using a macrocyclic design, to develop optimized menaquinone biosynthesis inhibitors as new antibacterials, and to develop protein-based and small-molecule inhibitors of E1 activating enzymes to probe their molecular mechanisms and biological functions in cancer. Systematic correlation of physicochemical properties with bacterial uptake will also be studied. This work will have broad impacts in structure-based design, antibacterial medicinal chemistry, and enzymology, and will be carried out through established multidisciplinary collaborations comprising combined expertise in organic synthesis, medicinal chemistry, pharmacology, biochemistry, microbiology, and structural biology.