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）：氨基糖苷类抗生素是用于治疗严重细菌感染的广谱抗生素，包括致命的结核病和伴随的艾滋病、囊性纤维化和癌症。对这些药物产生耐药性的病原体的出现是对公共卫生的重大威胁，并强调了对新的抗菌剂的需求。在目的1中，我们提出利用氨基糖苷乙酰转移酶（AAC）家族的氨基糖苷修饰酶结合新开发的无6 ′-N-酰化保护基的化学方法（i）来产生新的和更有效的N-酰化氨基糖苷抗生素的体外文库，和（ii）开发氨基糖苷类探针，其将用作鉴定这些抗生素的新的治疗性蛋白质靶点/途径的诱饵。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）目前还没有用于产生N-酰化氨基糖苷的通用合成方法，以及（ii）没有有效的方法来化学修饰含有一系列化学相同胺的氨基糖苷上的特定胺基。一些现有的例子，只使用化学合成太苛刻的研究时间和成本，并仅限于非常具体的情况。在目标2中，我们提出了广泛耐药的M.广泛耐药结核病。我们希望这项工作能够（i）推进对包括M.结核病，和（ii）提供了一个潜在的解决方案，以克服大多数广泛耐药结核病的氨基糖苷类耐药问题。与公共卫生的相关性：我们希望这项工作将有助于开发新的抗生素，具有对抗现有和新出现的耐药细菌的潜力。 公共卫生相关性：拟议的研究与公共卫生有关，因为对氨基糖苷类耐药酶的更好理解有望导致新药的发现和开发，并有助于确定药物靶点。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。该研究将在两个方面产生重大影响：（1）我们用于氨基糖苷类药物的快速和简便的区域选择性N-酰化的新型化学和化学酶方法将大大降低通常与此类修饰相关的成本和工作量，以及（2）我们对广泛耐药M中氨基糖苷类药物抗性主要决定因素的作用机制和抑制的研究。结核病的发现将促进我们对耐药性的理解，并在抗生素领域建立氨基糖苷乙酰转移酶的新范式。