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.

摘要 对常见的一线抗生素，如甲氧西林、万古霉素、头孢菌素， 和碳青霉烯类抗生素，已经为数百万患者带来了全球公共卫生挑战。因此，至关重要的是要发现和 商业化新的抗菌剂，可以成功地中和多药耐药细菌（MDRB）， 毒性本提案的目的是开发独特的一流的两亲性环状抗菌肽（AMP） 对临床相关病原体如屎肠球菌、金黄色葡萄球菌、克雷伯菌 肺炎杆菌、鲍曼不动杆菌、铜绿假单胞菌和肠杆菌属（ESKAPE病原体）。我们 建议开发含有天然和/或非天然疏水性和带正电荷残基的AMP，因为它们的宽- 使用体外和体内测定，对特定MDR病原体的谱活性和功效。我们发现 包含色氨酸（W）和精氨酸（R）氨基酸的环状两亲肽[R4 W 4]有效对抗 多种细菌病原体，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）、肺炎克雷伯菌（MIC = 16.0 µg/mL）和大肠埃希菌（MIC = 16.0µg/mL），并与四环素对MRSA显示出协同活性，与异烟肼和吡嗪酰胺对MRSA显示出协同活性。 结核分枝杆菌。基于此模板，我们产生了一个新的肽库（>200）， 抗菌活性。例如，IFX-027、IFX-135、IFX-145、IFX-146、IFX-154和IFX-301显示MIC = 1.5-25 μg/mL，对革兰氏阳性菌和革兰氏阴性菌。几种先导化合物显示出与以下化合物的协同活性： 其他几种抗生素的部分抑菌浓度（FIC）指数范围为0.3-0.5。我们的先导肽（IFX- 031、IFX-031-1和IFX-111）也减少MRSA和铜绿假单胞菌的生物膜形成。IFX-301被发现是 在小鼠中在50 mg/kg的剂量水平下是无毒的，并且所有肽对人红细胞（hRBC）都是无毒的。 (HC50>500 μg/mL）。在目标1中，我们将基于六个前导肽建立结构-活性关系（SAR）， 深入了解负责分子对细菌病原体的选择性的结构决定因素。的 大多数有效的化合物将进一步评估其稳定性、细胞毒性和随着时间的推移对抗菌剂的发展。 阻力目标1的拟议里程碑是确定MIC ≤5 µg/mL和MIC ≤10的5种先导肽类似物 对革兰氏阳性菌和革兰氏阴性菌分别为μg/mL，20倍浓度hRBC溶血≤5% MIC值。在目标2中，我们将评估体内功效和毒性、初步药代动力学（例如，Cmax，tmax， t1/2），以及目标1中鉴定的2-3种前导抗微生物肽类似物对鼠感染模型抗四种 致病菌在第一阶段成功完成时，具有大治疗指数的最有效化合物将 进入II期研究，并成为IND申请的重点。

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