A New Antibacterial Drug Target: Analyzing Inhibitor Binding to a Bacterial Metal

新的抗菌药物靶点：分析抑制剂与细菌金属的结合

基本信息

批准号：
7981091
负责人：
Daniel Paul Becker
金额：
$ 40.02万
依托单位：
LOYOLA UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-15 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7981091
关键词：
Active Sites Address Anti-Bacterial Agents Antibiotic Resistance Antibiotics Area Bacteria Binding Chicago Clinic Collaborations Data Diaminopimelic Acid Drug Delivery Systems Electron Spin Resonance Spectroscopy Enzyme Inhibitor Drugs Enzyme Inhibitors Enzymes Escherichia coli Exhibits Foundations Gene Deletion Goals Haemophilus influenzae Health Healthcare Helicobacter pylori Human Indiana Kinetics Laboratories Lead Lysine Mammals Metals Methods Mononuclear Multi-Drug Resistance Mycobacterium smegmatis Organism Pathway interactions Pharmaceutical Chemistry Property Research Resolution Roentgen Rays Social Welfare Societies Spectrum Analysis Structure Testing Toxic effect Translating Universities Wisconsin X-Ray Crystallography analog antimicrobial antimicrobial drug bacterial genetics bacterial resistance base cell growth combat design enthalpy functional group high throughput screening in vivo inhibitor/antagonist insight medical schools mutant novel pathogenic bacteria professor public health relevance resistant strain small molecule

项目摘要

DESCRIPTION (provided by applicant): The importance of this project is underscored by the emergence of several pathogenic bacterial strains that are resistant to all currently available antibiotics. One way to combat antibiotic resistance is to establish new enzymatic targets within resistant bacterial strains and design and synthesize small molecule inhibitors to target these enzymes. Based on bacterial genetic information, the meso-diaminopimelate (mDAP)/lysine biosynthetic pathway offers several potential anti-bacterial targets that have yet to be explored. Since there are no similar pathways in mammals, inhibitors that target one or more of the enzymes in this biosynthetic pathway will likely exhibit selective toxicity against only bacteria. It has been shown that deletion of the gene encoding for one of these enzymes, the dapE-encoded N-succinyl-L,L-diaminopimelic acid desuccinylase (DapE; EC 3.5.1.18), is lethal to Helicobacter pylori and Mycobacterium smegmatis. Even in the presence of lysine supplemented media H. pylori was unable to grow. Therefore, DapE9s are essential for cell growth and proliferation. A major limitation in developing a novel class of antibiotics that target DapE enzymes is the lack of X-ray crystallographic information. Recently, the PI in collaboration with Dr. Boguslaw Nocek at Argonne National Laboratory, solved the 2.0 and 2.3 E resolution structures of the mono and dinuclear Zn(II) DapE enzymes from the pathogenic bacterium Haemophilus influenzae. These structures provide the foundation needed to examine the binding of several new, potent inhibitors of DapE that were recently discovered by the PI, some of which have been shown to possess antimicrobial activity towards Escherichia coli. The specific aims of this proposal are: i) Discover novel DapE inhibitors by high throughput screening, ii) Analyze the determinants of substrate/inhibitor binding to DapE, and iii) Examine active site residues that are involved in substrate/inhibitor binding. Results obtained from these studies will identify new medicinal chemistry leads for optimization as well as mechanistic insight into the determinants of substrate recognition and binding. These data will provide insight into which functional groups are most important to increase inhibitor binding enthalpy to DapE. The most promising compounds discovered will be tested for antimicrobial properties in vivo against several pathogenic bacterial strains providing an avenue to translate newly discovered antimicrobial leads into the clinic. It is anticipated that the successful completion of the studies described in this proposal will provide benefits to healthcare and the general welfare of society given that multidrug-resistant organisms pose a serious and increasing treat to human health. PUBLIC HEALTH RELEVANCE: The importance of this project is underscored by the emergence of several pathogenic bacterial strains that are resistant to all currently available antibiotics. To address this problem, we propose to identify new medicinal chemistry leads for optimization and analyze the determinants of substrate/inhibitor binding to the dapE-encoded N- succinyl-L,L-diaminopimelic acid desuccinylase (DapE) from Haemophilus influenzae, a novel antimicrobial target that is apart of the lysine biosynthetic pathway in bacteria. The most promising compounds will be tested for antimicrobial properties in vivo against several pathogenic bacterial strains providing an avenue to translate newly discovered antimicrobial leads into the clinic.

描述（由申请人提供）：该项目的重要性强调了几种对所有当前可用抗生素具有抗性的致病细菌菌株的出现。打击抗生素耐药性的一种方法是在抗性细菌菌株中建立新的酶促靶标，并设计并合成小分子抑制剂以靶向这些酶。基于细菌遗传信息，中二氨基二亚二二二二酸酯（MDAP）/赖氨酸生物合成途径提供了尚未探索的几种潜在的抗细菌靶标。由于哺乳动物没有类似的途径，因此靶向该生物合成途径中一种或多种酶的抑制剂可能仅对细菌表现出选择性毒性。已经表明，编码这些酶之一的基因的缺失是DAPE代码编码的N-核酸-L-二氨基二酰胺酸脱甲基蛋白酶（Dape; EC 3.5.1.1.18），是杀虫剂幽门螺杆菌和分枝杆菌的杀伤力。即使在存在赖氨酸的存在下，幽门螺杆菌也无法生长。因此，DAPE9对于细胞生长和增殖至关重要。开发靶向DAPE酶的新型抗生素类型的主要局限性是缺乏X射线晶体学信息。最近，PI与Argonne National Laboratory的Boguslaw Nocek博士合作，解决了来自嗜嗜性嗜血杆菌的致病细菌的单核Zn（II）Dape酶的2.0和2.3 E分辨率结构。这些结构为检查了几种新的，有效的DAPE抑制剂的结合所需的基础，这些抑制剂最近被PI发现，其中一些已显示出对大肠杆菌具有抗菌活性。该提案的具体目的是：i）通过高吞吐量筛选发现新型的DAPE抑制剂，ii）分析底物/抑制剂与DAPE的决定因素，以及iii）检查与底物/抑制剂结合有关的活性位点残基。从这些研究中获得的结果将确定新的药物化学领导，以优化底物识别和结合的决定因素。这些数据将提供有关哪些功能组对于增加抑制剂结合焓与DAPE最重要的信息。发现的最有希望的化合物将在体内测试与几种致病细菌菌株的体内抗菌特性，从而提供了将新发现的抗菌铅转化为诊所的途径。预计该提案中描述的研究的成功完成将为医疗保健和社会的总体福利带来好处，因为多种耐药的生物对人类健康造成了严重且日益严重的治疗。公共卫生相关性：该项目的重要性强调了几种对所有当前可用抗生素具有抗性的致病细菌菌株的出现。为了解决这个问题，我们建议确定新的药物化学铅，以优化并分析与DAPE编码的N-琥珀酰-L-琥珀酰-L-二氨基二酰胺酸去甲酸脱糖苷酶（DAPE）的底物/抑制剂的决定因素。最有希望的化合物将在体内测试与几种致病细菌菌株的体内抗菌特性，从而提供了将新发现的抗菌铅转化为诊所的途径。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The dimerization domain in DapE enzymes is required for catalysis.

DOI：
10.1371/journal.pone.0093593
发表时间：
2014
期刊：
PloS one
影响因子：
3.7
作者：
Nocek B;Starus A;Makowska-Grzyska M;Gutierrez B;Sanchez S;Jedrzejczak R;Mack JC;Olsen KW;Joachimiak A;Holz RC
通讯作者：
Holz RC

Mono-N-acyl-2,6-diaminopimelic acid derivatives: analysis by electromigration and spectroscopic methods and examination of enzyme inhibitory activity.

单-N-酰基-2,6-二氨基庚二酸衍生物：通过电迁移和光谱方法进行分析以及酶抑制活性的检查。

DOI：
10.1016/j.ab.2014.08.032
发表时间：
2014
期刊：
Analytical biochemistry
影响因子：
2.9
作者：
Hlaváček,Jan;Vítovcová,Miloslava;Sázelová,Petra;Pícha,Jan;Vaněk,Václav;Buděšínský,Miloš;Jiráček,Jiří;Gillner,DanutaM;Holz,RichardC;Mikšík,Ivan;Kašička,Václav
通讯作者：
Kašička,Václav

Identification of a Histidine Metal Ligand in the argE-Encoded N-Acetyl-L-Ornithine Deacetylase from Escherichia coli.

DOI：
10.1186/2193-1801-2-482
发表时间：
2013
期刊：
SpringerPlus
影响因子：
0
作者：
McGregor WC;Gillner DM;Swierczek SI;Liu D;Holz RC
通讯作者：
Holz RC

Immobilization of the aminopeptidase from Aeromonas proteolytica on Mg2+/Al3+ layered double hydroxide particles.

DOI：
10.1021/am1005095
发表时间：
2010-10
期刊：
ACS APPLIED MATERIALS & INTERFACES
影响因子：
9.5
作者：
Frey, Steven T.;Guilmet, Stephanie L.;Egan, Richard G., III;Bennett, Alyssa;Soltau, Sarah R.;Holz, Richard C.
通讯作者：
Holz, Richard C.