Disrupting Dogma: Investigating LPS Biosynthesis Inhibition as an Alternative Mechanism of Action of Aminoglycoside Antibiotics

颠覆教条：研究 LPS 生物合成抑制作为氨基糖苷类抗生素的替代作用机制

基本信息

批准号：
10653587
负责人：
Erika A Taylor
金额：
$ 47.47万
依托单位：
WESLEYAN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10653587
关键词：
Active Sites Affinity Amines Aminoglycoside Antibiotics Aminoglycosides Anti-Bacterial Agents Antibiotic Resistance Antibiotics Aptitude Bacterial Infections Big Data Big Data Methods Binding Binding Sites Biochemical Biochemistry Biological Assay Biophysics Calcium Channel Cations Cell Fraction Cell membrane Cells Charge Chemistry Collaborations Computing Methodologies Data Development Docking Drug Design Ensure Enzymes Escherichia coli Escherichia coli Proteins Evaluation Event Fluorescence Future Gel Chromatography Goals Gram-Negative Bacteria Human In Vitro Investigation Kinetics Knowledge Libraries Ligand Binding Lipopolysaccharide Biosynthesis Pathway Mass Spectrum Analysis Methods Microbial Drug Resistance Modification Morbidity - disease rate Mutagenesis Permeability Pharmaceutical Preparations Phenotype Phosphorylases Protein Analysis Protein Biosynthesis Proteins RNA chemical synthesis Research Ribosomes Science Spectrum Analysis Statistical Data Interpretation Statistical Methods Structure-Activity Relationship Student recruitment Students Testing Toxic effect Training Universities Work X-Ray Crystallography amidase bactericide cellular targeting chemical group cohort comparative computer studies design drug discovery effective therapy experimental study guanidinium in silico inhibitor intermolecular interaction molecular dynamics molecular recognition mortality nanomolar nephrotoxicity new therapeutic target novel ototoxicity priority pathogen protein expression protein function protein structure response side effect structural biology student training success synergism undergraduate student

项目摘要

Project Summary With numerous Gram-negative bacterial species demonstrating antimicrobial drug resistance, the identification of new inhibitors and the optimization of existing inhibitors is necessary to enable an effective treatment of illnesses. Recent research efforts in our lab and others have demonstrated that aminoglycosides have minimal impact on protein synthesis and in fact they potently bind to heptosytransferase I (HepI) in Escherichia coli. This is an important finding, because it may allow for this class of antibiotics to be dramatically redesigned to be better drugs with fewer side effects, because the HepI and ribosome binding sites have very different sizes and they have dramatic differences in charges (HepI is positively charged, while the ribosome is negatively charged). This proposal will advance efforts to redesign aminoglycoside antibiotics to enhance HepI binding and to reduce binding to other cellular targets that can lead to side effects like oto- and nephrotoxicity. Our investigation will address three hypotheses: (1) that aminoglycosides bind to other cellular targets beyond the ribosome including heptosyltransferase enzymes, (2) understanding the HepI-aminoglycoside interactions will enable structural modification and optimization of bactericidal activity, and (3) that computational methods can enhance aminoglycoside redesign. To date, efforts to redesign aminoglycosides for more potent binding to the ribosome has failed to lead to more potent drugs, and this is likely because the mechanism of action involves other enzymes like HepI. This work promises to enhance drug discovery efforts while also providing training for students in my lab and in two upper-level biochemistry courses at Wesleyan in 21st century drug discovery methods.

项目概要由于许多革兰氏阴性细菌表现出抗菌药物耐药性，因此鉴定新抑制剂的开发和现有抑制剂的优化对于有效治疗疾病。我们实验室和其他实验室最近的研究工作表明，氨基糖苷类药物具有最小的对蛋白质合成的影响，事实上它们可以有效地与大肠杆菌中的庚基转移酶 I (HepI) 结合。这这是一个重要的发现，因为它可能允许对此类抗生素进行大幅重新设计，使其变得更好副作用较少的药物，因为 HepI 和核糖体结合位点的大小非常不同，并且它们电荷有显着差异（HepI 带正电，而核糖体带负电）。这该提案将推进重新设计氨基糖苷类抗生素的努力，以增强 HepI 结合并减少与其他细胞靶标结合可能导致耳毒性和肾毒性等副作用。我们的研究将解决三个假设：（1）氨基糖苷类药物除了与其他细胞靶标结合外，还与其他细胞靶标结合。核糖体，包括庚糖基转移酶，(2) 了解 HepI-氨基糖苷相互作用将能够实现结构修饰和杀菌活性优化，并且（3）计算方法可以增强氨基糖苷类药物的重新设计。迄今为止，为了更有效地结合氨基糖苷类药物，人们正在努力重新设计氨基糖苷类药物。核糖体未能产生更有效的药物，这可能是因为作用机制涉及其他酶如 HepI。这项工作有望加强药物发现工作，同时还提供培训我实验室的学生以及卫斯理学院两门关于 21 世纪药物发现的高级生物化学课程的学生方法。