Membrane-active quinoline and quinazoline antibacterials that target Gram positive pathogens

针对革兰氏阳性病原体的膜活性喹啉和喹唑啉抗菌剂

基本信息

批准号：
10117071
负责人：
Ambrose Lin Yau Cheung
金额：
$ 76.46万
依托单位：
DARTMOUTH COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10117071
关键词：
Affinity Chromatography Animal Model Anti-Bacterial Agents Antibiotic Therapy Antibiotics Bacteria Binding Binding Proteins Biological Assay Cells Daptomycin Data Defect Drug Kinetics Dyes Electron Microscopy Endocarditis Ensure Enterococcus Enterococcus faecalis Enterococcus faecium Evaluation Studies Exhibits Frequencies Genetic Genomics Goals Gram-Positive Bacteria Hospitals In Vitro Infection Infection Control Infectious Skin Diseases Infective endocarditis Lead Length of Stay Libraries Linezolid Lipid Bilayers Lipids Mass Spectrum Analysis Measures Membrane Minnesota Modeling Modification Molecular Multi-Drug Resistance Mus Mutation New Agents Nosocomial Infections Organism Oryctolagus cuniculus Pathogenicity Pharmaceutical Preparations Plasma Proteins Positioning Attribute Procedures Property Proteomics Public Health Publishing Quinazolines Quinolones Research Resistance Resort Safety Series Serum Solubility Specificity Staphylococcus aureus infection Staphylococcus epidermidis Streptococcus pneumoniae Structure Structure-Activity Relationship Toxic effect Universities Vancomycin Work activity-based protein profiling analog aqueous bactericide base biochemical tools biophysical tools carbonyl group crosslink cytotoxicity design drug development drug discovery drug disposition efficacy evaluation efficacy study improved in vivo lipophilicity medical schools methicillin resistant Staphylococcus aureus mutant novel pathogen quinoline scaffold screening simulation

项目摘要

Abstract Infections due to resistant Gram+ organisms are on the rise, likely due to a variety of factors including longer hospital stay, increased frequency of invasive procedures and pervasive antibiotic therapy. Compounding the problem is the emergence of multi-drug resistance (MDR) among many Gram+ pathogens (MRSA, S. epidermidis, Enterococcus and S. pneumoniae). Despite antibiotic stewardship and infection control, new agents against these Gram+ pathogens are urgently needed. After screening a ~60,000 preselected compound library, we obtained DNAC-2, a 4-hydroxyquinoline derivative, that exhibited antibacterial activities against MRSA and Enterococcus. We subsequently synthesized 3 series of analogues involving over 50 compounds. Two of these analogues in the 2th series, JRS-3-56 (compound 1) and JRS-4-32 (compound 2), were cidal against MRSA, S. epidermidis, E. faecalis and E. faecium, with MIC ≤0.2 μg/ml. However, both 1 and 2 have poor predicted aqueous solubility with high cLogP (7.7 and 6.0, respectively). Conversion of quinoline to quinazoline for 1 improved the cLogP (from 6.06 to 5.08) but led to a slight increase in MIC (0.25 to 2 µg/ml). In the latest series, we introduced a carbonyl group at C-4 and a C to N substitution at the C-1 position, yielding compounds 3 and 4 with low cLogPs and very low MIC (0.06 µg/ml for USA300), accompanied by a much tighter SAR. Using macromolecular synthesis assays, membrane-specific dye FM4- 64 and electron microscopy studies, we have evidence that 1 and 2 target the Gram+ membrane (3 and 4 also resulted in membrane defect as detected by the FM4-64 dye), but not Gram- or eukaryotic membrane, thus implying some degree of specificity. However, the exact target of these compounds which likely differs from daptomycin, is not known. In this application, we seek to define the mechanism of action of these quinoline/quinolone derivatives and further explore the SAR that governs in vitro and in vivo activities and drug disposition properties. Accordingly, we have the following specific aims: 1) design and synthesize quinoline/quinolone derivates by defining the SAR that governs activity against major Gram+ pathogens and drug disposition properties (MIC, solubility, overt toxicity and serum binding etc.); 2) delineate the mechanism of action of the quinoline/quinolone derivatives with genetic, biochemical and biophysical tools; 3) pharmacokinetic and efficacy studies where candidates compounds will be evaluated for their drug disposition properties to ensure safety and selectivity followed by selection of “lead” compounds for full PK evaluation and efficacy studies with two animal models. The goal of these studies is to identify “druggable membrane-active compounds” with broad Gram+ activity. If successful, we believe these compounds will represent a new class of membrane-active compounds that offer a significant advance in drug development.

抽象的由耐药革兰氏阳性菌引起的感染呈上升趋势，这可能是由于多种因素造成的，包括较长时间住院时间、侵入性手术频率增加和普遍的抗生素治疗。问题是许多革兰氏阳性病原体（MRSA、S. 尽管有抗生素管理和感染控制，但仍存在新的感染。在筛选了约 60,000 种预选后，迫切需要针对这些革兰氏阳性病原体的药物。化合物库中，我们获得了DNAC-2，一种4-羟基喹啉衍生物，具有抗菌活性随后我们合成了 3 个系列的类似物，涉及 50 多种。第二系列中的两个类似物，JRS-3-56（化合物1）和JRS-4-32（化合物2），对 MRSA、表皮葡萄球菌、粪肠球菌和屎肠球菌均具有杀灭作用，MIC ≤0.2 μg/ml 然而，两者均 1。 1 和 2 的预测水溶性较差，且 cLogP 的转化率分别为 7.7 和 6.0。喹啉至喹唑啉 1 改善了 cLogP（从 6.06 至 5.08），但导致 MIC 略有增加（0.25 在最新系列中，我们在 C-4 处引入了羰基，并在 C-1 处引入了 C 至 N 取代。位置，产生具有低 cLogP 和极低 MIC（USA300 为 0.06 µg/ml）的化合物 3 和 4，伴随着更严格的 SAR 使用大分子合成、膜特异性染料 FM 测定4- 64 和电子显微镜研究，我们有证据表明 1 和 2 靶向 Gram+ 膜（3 和 4 也导致了 FM4-64 染料检测到的膜缺陷），但不是革兰氏或真核细胞膜，因此意味着一定程度的特异性。然而，这些化合物的确切目标可能不同。达托霉素的作用尚不清楚，在本申请中，我们试图确定这些药物的作用机制。喹啉/喹诺酮衍生物并进一步探索控制体外和体内活性的SAR 因此，我们有以下具体目标：1）设计和合成。喹啉/喹诺酮衍生物，通过定义控制主要革兰氏+病原体活性的SAR 药物处置特性（MIC、溶解度、明显毒性和血清结合等）；2）描述机制；喹啉/喹诺酮衍生物与遗传、生物化学和生物物理工具的作用；3) 药代动力学和功效研究，其中候选化合物将被评估其药物处置特性，以确保安全性和选择性，然后选择“先导”化合物进行全面 PK 使用两种动物模型进行评估和功效研究，这些研究的目的是确定“可成药”。具有广泛革兰氏+活性的膜活性化合物”如果成功，我们相信这些化合物将会实现。一类新型膜活性化合物，为药物开发带来了重大进展。