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，包括致命的结核病以及随附的艾滋病，囊性纤维化和癌症。病原体对这些药物具有抗性的出现代表了对公共卫生的主要威胁，并强调了对新的抗菌剂的需求。在AIM 1中，我们建议利用氨基糖苷修饰酶的酶乙酰基转移酶（AAC）家族以及新开发的6'-N-囊泡保护群体无群的化学方法（I），以生成新的和更有效的n- acty to an-acy to an-acy an-acy to an-acy to an-acy to an-acy to an-acy to an-acy to andyminogibibibibibib，氨基糖苷探针将用作鉴定这些抗生素的新型治疗蛋白靶/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）没有现有的一般合成方法来创建N-酰基化的氨基糖苷，并且（ii）没有有效修饰特定胺基的氨基糖苷上的特定胺的方法，其中包含一系列化学上相同的胺。一些仅使用化学合成的现有示例对研究时间和成本的要求太高，并且仅限于非常具体的情况。在AIM 2中，我们提出了对作用机理的生化和结构研究，并抑制了结核分枝杆菌（XDR-TB）的广泛药物抗药性菌株中氨基糖苷耐药性的主要决定因素。我们希望这项工作（i）提高对各种致病细菌（包括结核分枝杆菌）的Ag耐药性的基本理解，并且（ii）提供了一种潜在的解决方案来克服大多数XDR-TB的氨基糖苷耐药性问题。与公共卫生有关：我们希望这项工作将有助于开发新型抗生素，并有可能抵抗现有和新兴的耐药菌细菌。 公共卫生相关性：拟议的研究与公共卫生有关，因为人们对氨基糖苷 - 抗性酶有了深入的了解，预计将导致发现和开发新药以及识别药物靶标。这项研究将使我们能够更好地解决感染性细菌对氨基糖苷抗生素的耐药性问题。拟议的研究将以两种方式产生重大影响：（1）我们的新型化学和化学酶方法学，用于快速且便捷的氨基糖苷的快速和便捷的选择性n-酰化，从而大大降低与这种修改相关的成本和努力，以及（2）我们对Aminogycsdie Incorsistriant tuber interive Incoristion to and Incoristion to andie contimistion to andie consistion tubor intercristion的研究，促进我们对耐药性的理解，还为抗生素领域中的氨基糖苷乙酰转移酶树立了新的范式。