Chemoenzymatic studies of aminoglycoside-resistance enzymes towards new drugs

新药氨基糖苷类耐药酶的化学酶学研究

• 批准号: 8185756
• 负责人: Sylvie Garneau-Tsodikova
• 金额: $ 45.77万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8185756
• 关键词: Acetylation; Acetyltransferase; Acquired Immunodeficiency Syndrome; Acute; Acyl Coenzyme A; Affinity; Amines; Aminoglycoside Antibiotics; Aminoglycoside resistance; Aminoglycosides; Anti-Bacterial Agents; Antibiotics; Bacillus anthracis; Bacteria; Bacterial Infections; Biochemical; Biological; Chemicals; Chemistry; Clinic; Clinical; Coenzyme A; Collaborations; Complement; Complex; Cystic Fibrosis; Development; Disease; Drug Delivery Systems; Drug resistance; Drug resistance in tuberculosis; Enterococcus faecalis; Enzymes; Extreme drug resistant tuberculosis; Family; Health; Homologous Gene; Human; In Vitro; Kanamycin A; Kinetics; Klebsiella pneumonia bacterium; Lead; Libraries; Malignant Neoplasms; Methodology; Methods; Michigan; Modification; Multi-Drug Resistance; Mycobacterium tuberculosis; N acylation; One-Step dentin bonding system; Parents; Pathway interactions; Pharmaceutical Preparations; Positioning Attribute; Proteins; Public Health; Research; Resistance; Resistance development; Ribosomes; Series; Solutions; Structure; Testing; Therapeutic; Time; Toxic effect; Tuberculosis; Universities; Work; aminoglycoside acetyltransferase; analog; antimicrobial drug; bacterial resistance; chemical synthesis; combat; cost; drug resistant bacteria; fascinate; improved; inhibitor/antagonist; innovation; insight; novel; novel therapeutics; pathogen; pathogenic bacteria; prevent; resistance mechanism; resistant strain; therapeutic protein

DESCRIPTION (provided by applicant): Aminoglycosides are broad-spectrum antibiotics used for treatment of serious bacterial infections, including the deadly tuberculosis and those accompanying AIDS, cystic fibrosis, and cancer. The emergence of pathogens resistant to these drugs represents a major threat to public health and underscores the need for new antimicrobial agents. In Aim 1, we propose to utilize aminoglycoside-modifying enzymes of the aminoglycoside acetyltransferase (AAC) family in conjunction with a newly developed 6'-N-acylation protecting group-free chemical methodology (i) to generate in vitro libraries of new and more potent N-acylated aminoglycoside antibiotics, and (ii) to develop aminoglycoside probes that will serve as baits for identification of novel therapeutic protein targets/pathways for these antibiotics. Our chemoenzymatic and chemical strategies offer an effective solution to the following problems: (i) there are no existing general synthetic methodologies for the creation of N-acylated aminoglycosides, and (ii) there are no efficient methods to chemically modify specific amine groups on an aminoglycoside that contains a series of chemically identical amines. A few existing examples that use solely chemical syntheses are too demanding on research time and cost and are limited to very specific cases. In Aim 2, we propose biochemical and structural studies of the mechanism of action and inhibition of a major determinant of aminoglycoside resistance in extensively drug-resistant strains of M. tuberculosis (XDR-TB). We expect this work to (i) advance the basic understanding of AG resistance of a variety of pathogenic bacteria, including M. tuberculosis, and (ii) provide a potential solution to overcome the aminoglycoside resistance problem in majority of XDR- TB. Relevance to public health: We expect that this work will contribute to the development of novel antibiotics with a potential to combat existing and newly emerging drug-resistant bacteria. PUBLIC HEALTH RELEVANCE: The proposed research is relevant to public health as an improved understanding of aminoglycoside- resistance enzymes is expected to lead to the discovery and development of new drugs and also in identifying drug targets. This research will enable us to better handle the problem of resistance of infectious bacteria to aminoglycoside antibiotics. The proposed research will have a major impact in two ways: (1) our novel chemical and chemoenzymatic methodologies for the rapid and facile regio- selective N-acylation of aminoglycosides will greatly reduce the cost and effort generally associated with such modifications, and (2) our study of the mechanism of action and inhibition of the major determinant of aminoglycosdie resistance in extensively drug-resistant M. tuberculosis will advance our understanding of drug resistance and also set a new paradigm of aminoglycoside acetyltransferase in the antibiotics field.

描述（由申请人提供）：氨基糖苷类是广谱抗生素，用于治疗严重的细菌感染，包括致命的结核病以及伴随艾滋病、囊性纤维化和癌症的感染。对这些药物产生耐药性的病原体的出现对公众健康构成了重大威胁，并强调了对新抗菌药物的需求。在目标 1 中，我们建议利用氨基糖苷乙酰转移酶 (AAC) 家族的氨基糖苷修饰酶与新开发的 6'-N-酰化无保护基化学方法相结合 (i) 生成新的、更有效的 N-酰化氨基糖苷类抗生素的体外文库，以及 (ii) 开发 氨基糖苷类探针将作为诱饵来鉴定这些抗生素的新型治疗蛋白靶点/途径。我们的化学酶和化学策略为以下问题提供了有效的解决方案：(i) 没有现有的通用合成方法来创建 N-酰化氨基糖苷，以及 (ii) 没有有效的方法对包含一系列化学相同胺的氨基糖苷上的特定胺基进行化学修饰。一些仅使用化学合成的现有例子对研究时间和成本要求过高，并且仅限于非常具体的情况。在目标 2 中，我们建议对广泛耐药结核分枝杆菌 (XDR-TB) 菌株中氨基糖苷类耐药的主要决定因素的作用和抑制机制进行生化和结构研究。我们期望这项工作能够（i）增进对包括结核分枝杆菌在内的多种病原菌的 AG 耐药性的基本了解，以及（ii）为克服大多数广泛耐药结核病的氨基糖苷类耐药性问题提供潜在的解决方案。与公共卫生的相关性：我们期望这项工作将有助于新型抗生素的开发，这些抗生素有可能对抗现有和新出现的耐药细菌。 公共健康相关性：拟议的研究与公共健康相关，因为对氨基糖苷类耐药酶的进一步了解预计将导致新药的发现和开发以及药物靶点的确定。这项研究将使我们能够更好地处理感染性细菌对氨基糖苷类抗生素的耐药性问题。拟议的研究将在两个方面产生重大影响：(1)我们用于氨基糖苷类快速、简便的区域选择性N-酰化的新型化学和化学酶方法将大大减少通常与此类修饰相关的成本和工作量，(2)我们对广泛耐药结核分枝杆菌中氨基糖苷耐药性的主要决定因素的作用和抑制机制的研究将 增进了我们对耐药性的理解，并在抗生素领域树立了氨基糖苷乙酰转移酶的新范例。