Development of Broad-Spectrum Cyclic Amphiphilic Peptides against Multidrug-Resistant Bacteria

抗多重耐药细菌的广谱环状两亲肽的开发

• 批准号: 10685928
• 负责人: Assad Kazeminy
• 金额: $ 29.66万
• 依托单位: AJK BIOPHARMACEUTICAL LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-18 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10685928
• 关键词: Acinetobacter baumannii; Address; Adult; Amino Acids; Anti-Bacterial Agents; Antibiotics; Antimicrobial Resistance; Arginine; Bacteria; Bacterial Antibiotic Resistance; Bacterial Drug Resistance; Biological Assay; Body Weight; C57BL/6 Mouse; Carbapenems; Cell membrane; Centers for Disease Control and Prevention (U.S.); Cephalosporins; Charge; Cyclic Peptides; Daptomycin; Data; Development; Dose; Drug Kinetics; Drug resistance; ESKAPE pathogens; Enterobacter; Enterococcus faecium; Erythrocytes; Escherichia coli; Eukaryotic Cell; Exhibits; Feasibility Studies; Government; Half-Life; Health Personnel; Hemolysis; Histology; Human; Hydrophobicity; In Vitro; Inbred BALB C Mice; Infection; Intravenous; Klebsiella pneumoniae; Lead; Legal patent; Levaquin; Libraries; Lipids; Maximum Tolerated Dose; Medical; Membrane; Methicillin; Microbial Biofilms; Modeling; Monitor; Multi-Drug Resistance; Multiple Bacterial Drug Resistance; Mus; Mycobacterium tuberculosis; Patients; Peptide Library; Peptides; Periodicity; Persons; Phase; Polymyxin B; Pseudomonas aeruginosa; Public Health; Pyrazinamide; Reporting; Resistance; Resistance development; Safety; Staphylococcus aureus; Staphylococcus aureus infection; Structure-Activity Relationship; Tetracyclines; Therapeutic Index; Time; Toxic effect; Tryptophan; United Kingdom; Vancomycin; Work; amphiphilicity; antimicrobial; antimicrobial drug; antimicrobial peptide; antimicrobial resistant infection; assay development; biological systems; clinically relevant; cohort; commercialization; cytotoxicity; design; dosage; economic impact; efficacy study; experimental study; improved; in vivo; indexing; insight; isoniazid; lead optimization; methicillin resistant Staphylococcus aureus; mortality; mouse model; multi-drug resistant pathogen; novel; pathogen; pathogenic bacteria; peptide amphiphiles; peptide analog; phase 2 study; resistant strain; structural determinants

ABSTRACT The emergence of antibacterial resistance to common frontline antibiotics, such as methicillin, vancomycin, cephalosporins, and carbapenem, have created a global public health challenge for millions of patients. It is therefore critical to discover and commercialize new antimicrobial agents that can successfully neutralize multidrug-resistant bacteria (MDRB) with minimal toxicity. The objective of this proposal is to develop unique first-in-class amphiphilic cyclic antimicrobial peptides (AMPs) that are active against clinically relevant pathogens like Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species (ESKAPE pathogens). We propose to develop AMPs containing natural and/or unnatural hydrophobic and positively charged residues for their broad- spectrum activity and efficacy against specific MDR pathogens, using in vitro and in vivo assays. We have discovered that a cyclic amphipathic peptide [R4W4], which comprises tryptophan (W) and arginine (R) amino acids was effective against diverse bacterial pathogens, such as methicillin-resistant Staphylococcus aureus (MRSA) (MIC = 2.7 µg/mL), Pseudomonas aeruginosa (MIC = 42.8 µg/mL), Klebsiella pneumoniae (MIC = 16.0 µg/mL), and Escherichia coli (MIC = 16.0 µg/mL) and showed synergistic activity with tetracycline against MRSA, and isoniazid and pyrazinamide against Mycobacterium tuberculosis. Based on this template, we generated a new library of peptides (>200) with enhanced antimicrobial activities. For example, IFX-027, IFX-135, IFX-145, IFX-146, IFX-154, and IFX-301 showed MIC = 1.5-25 µg/mL against Gram+ve and Gram-ve bacteria. Several of the lead compounds demonstrated synergistic activity with several other antibiotics with fractional inhibitory concentration (FIC) indices ranging from 0.3-0.5. Our lead peptides (IFX- 031, IFX-031-1, and IFX-111) also reduced biofilm formation by MRSA and P. aeruginosa. IFX-301 was found to be nontoxic at a dose level of 50 mg/kg in mice, and all peptides were not toxic against human red blood cells (hRBC) (HC50>500 μg/mL). In Aim 1, we will establish a structure-activity relationship (SAR) based on the six lead peptides to obtain insights into the structural determinants responsible for the molecules’ selectivity towards bacterial pathogens. The most potent compounds will be further evaluated for their stability, cytotoxicity, and development over time to antimicrobial resistance. The proposed milestones for Aim 1 are to identify five lead peptide analogs with MIC ≤5 µg/mL and MIC ≤10 µg/mL respectively against Gram+ve and Gram-ve bacteria, and hRBC hemolysis of ≤5% at a concentration of 20 times the MIC value. In Aim 2, we will evaluate the in vivo efficacy and toxicity, preliminary pharmacokinetics (e.g., Cmax, tmax, t1/2), and efficacy of the 2-3 lead antimicrobial peptide analogs identified in Aim 1 on a murine infection model against four pathogenic bacteria. At the successful completion of Phase I, the most potent compound with a large therapeutic index will be advanced to Phase II studies and be the focus for an IND application.

抽象的 对普通前线抗生素的抗菌耐药性的出现，例如甲氧西林，万古霉素，头孢菌素， 和Carbapenem为数百万患者带来了全球公共卫生挑战。因此，发现和 商业化可以成功中和抗多药细菌（MDRB）的新抗菌剂 毒性。该提案的目的是开发独特的一流的两亲性循环抗微生物胡椒（AMPS） 对临床相关的病原体（如肠球菌，金黄色葡萄球菌，克雷伯菌）的活性活跃 肺炎，baumannii，铜绿假单胞菌和肠杆菌（Eskape病原体）。我们 提议开发含有天然和/或不自然疏水的AMP，并为其广泛的残留物带来积极充电的残留物 使用体外和体内测定法对特定MDR病原体的频谱活性和效率。我们发现 循环两亲肽[R4W4]，包括色氨酸（W）和精氨酸（R）氨基酸是有效的 潜水细菌病原体，例如耐甲氧西林金黄色葡萄球菌（MRSA）（MIC = 2.7 µg/ml）， 铜绿假单胞菌（MIC = 42.8 µg/ml），克雷伯菌肺炎（MIC = 16.0 µg/ml）和大肠杆菌（MIC = MIC = 16.0 µg/ml），与针对MRSA的四环素表现出协同活性，以及​​异oni氮和吡嗪酰胺针对 结核分枝杆菌。基于此模板，我们生成了一个新的Petides库（> 200），并增强了 抗菌活性。例如，IFX-027，IFX-135，IFX-145，IFX-146，IFX-154和IFX-301显示了MIC = 1.5-25 µg/ml针对革兰氏+VE和革兰氏阴性细菌。几种铅化合物表现出与 其他几种具有分数抑制浓度（FIC）指数的抗生素范围为0.3-0.5。我们的领导者petides（ifx- 031，IFX-031-1和IFX-111）也通过MRSA和铜绿假单胞菌降低了生物膜的形成。发现IFX-301是 小鼠50 mg/kg的剂量水平无毒，所有肽对人红细胞没有毒性（HRBC） （HC50>500μg/ml）。在AIM 1中，我们将基于六个铅宠物建立结构性关系（SAR） 获取对负责分子对细菌病原体的选择性的结构决定者的见解。 随着时间的流逝，大多数潜在化合物将进一步评估其稳定性，细胞毒性和发育 反抗。 AIM 1的提议的里程碑是识别五个具有MIC≤5µg/mL的铅肽类似物和MIC≤10 µg/mL分别针对革兰氏+VE和革兰氏阴性细菌，HRBC溶血≤5％，浓度为20倍 麦克风值。在AIM 2中，我们将评估初步药代动力学的体内效率和毒性（例如CMAX，TMAX， T1/2），以及2-3铅抗菌肽类似物的效率 致病细菌。成功完成第一阶段时，具有大型治疗指数的最潜在化合物将 将其提高到II期研究，并成为IND应用的重点。

项目成果

Membrane-Active Cyclic Amphiphilic Peptides: Broad-Spectrum Antibacterial Activity Alone and in Combination with Antibiotics.

• DOI: 10.1021/acs.jmedchem.2c01469
• 发表时间: 2022-12-08
• 期刊: Journal of medicinal chemistry
• 影响因子: 7.3
• 作者: Mohammed EHM;Lohan S;Ghaffari T;Gupta S;Tiwari RK;Parang K
• 通讯作者: Parang K

Structure-Based Rational Design of Small α-Helical Peptides with Broad-Spectrum Activity against Multidrug-Resistant Pathogens.

• DOI: 10.1021/acs.jmedchem.2c01708
• 发表时间: 2023-01-12
• 期刊: JOURNAL OF MEDICINAL CHEMISTRY
• 影响因子: 7.3
• 作者: Lohan, Sandeep;Konshina, Anastasia G.;Efremov, Roman G.;Maslennikov, Innokentiy;Parang, Keykavous
• 通讯作者: Parang, Keykavous