Phospholipid antimetabolite lipid ether amines for topical treatment of chronic wounds and associated biofilms.

磷脂抗代谢物脂醚胺，用于局部治疗慢性伤口和相关生物膜。

基本信息

批准号：
10384660
负责人：
DANIEL J GIBSON
金额：
$ 29.97万
依托单位：
INTEGUMED, LLC
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-10 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10384660
关键词：
Acinetobacter baumannii Acute Affect Affinity Alabama Amines Amputation Antibiotic Resistance Antibiotic Therapy Antimetabolites Antimicrobial Effect Area Bacteria Bacterial Adhesins Bacterial Antibiotic Resistance Bacterial Infections Bilateral Binding Bioinformatics Biological Assay Blood Glucose Cell membrane Cells Clinical Cluster Analysis Colony-forming units Confocal Microscopy Cutaneous Data Dermis Development Diabetes Mellitus Diabetic Foot Ulcer Diabetic mouse Diffuse Docking Dose Drug resistance ESKAPE pathogens Endotoxins Enterobacter Enterococcus faecium Enzyme-Linked Immunosorbent Assay Ethers Exotoxins Extracellular Matrix Family suidae Fluorescence Formulation Functional disorder Goals Growth Health Healthcare Healthcare Systems High Pressure Liquid Chromatography Histology Human Image Impaired wound healing In Situ In Vitro Inflammation Inflammatory Insulin Keratin Kidney Diseases Klebsiella pneumoniae Lead Ligands Lipase Lipids Lower Extremity Machine Learning Mammals Mass Spectrum Analysis Membrane Metabolic dysfunction Microbial Biofilms Modeling Modification Mus Mutation Obesity Outcome Measure Pathogenicity Pathway interactions Patients Phase Phospholipase Phospholipids Plasma Plasmids Polysaccharides Property Proteins Pseudomonas aeruginosa Pseudomonas aeruginosa infection Punch Biopsy Resistance Resolution Safety Sampling Signal Transduction Skin Skin wound healing Stains Staphylococcus aureus Sterile coverings Supervision Systems Biology Technology Testing Therapeutic Therapeutic Effect Thick Topical application Toxic effect Universities Vascular Diseases Virulence Virulence Factors Wound models amphiphilicity analog antimicrobial bacterial metabolism bactericide candidate selection chronic infection chronic wound clinically relevant comorbidity conditioning culture plates cytokine cytotoxic density diabetic ulcer high risk in vivo in vivo Model in vivo imaging involucrin knock-down lead candidate lipid metabolism lipid nanoparticle lipidome lipidomics metabolomics microbial microbicide migration mouse model novel pathogen primary outcome receptor resistance mechanism secondary outcome skin regeneration standard of care treatment effect wound wound biofilm wound closure wound dressing wound healing wound treatment

Acinetobacter baumannii Acute Affect Affinity Alabama Amines Amputation Antibiotic Resistance Antibiotic Therapy Antimetabolites Antimicrobial Effect Area Bacteria Bacterial Adhesins Bacterial Antibiotic Resistance Bacterial Infections Bilateral Binding Bioinformatics Biological Assay Blood Glucose Cell membrane Cells Clinical Cluster Analysis Colony-forming units Confocal Microscopy Cutaneous Data Dermis Development Diabetes Mellitus Diabetic Foot Ulcer Diabetic mouse Diffuse Docking Dose Drug resistance ESKAPE pathogens Endotoxins Enterobacter Enterococcus faecium Enzyme-Linked Immunosorbent Assay Ethers Exotoxins Extracellular Matrix Family suidae Fluorescence Formulation Functional disorder Goals Growth Health Healthcare Healthcare Systems High Pressure Liquid Chromatography Histology Human Image Impaired wound healing In Situ In Vitro Inflammation Inflammatory Insulin Keratin Kidney Diseases Klebsiella pneumoniae Lead Ligands Lipase Lipids Lower Extremity Machine Learning Mammals Mass Spectrum Analysis Membrane Metabolic dysfunction Microbial Biofilms Modeling Modification Mus Mutation Obesity Outcome Measure Pathogenicity Pathway interactions Patients Phase Phospholipase Phospholipids Plasma Plasmids Polysaccharides Property Proteins Pseudomonas aeruginosa Pseudomonas aeruginosa infection Punch Biopsy Resistance Resolution Safety Sampling Signal Transduction Skin Skin wound healing Stains Staphylococcus aureus Sterile coverings Supervision Systems Biology Technology Testing Therapeutic Therapeutic Effect Thick Topical application Toxic effect Universities Vascular Diseases Virulence Virulence Factors Wound models amphiphilicity analog antimicrobial bacterial metabolism bactericide candidate selection chronic infection chronic wound clinically relevant comorbidity conditioning culture plates cytokine cytotoxic density diabetic ulcer high risk in vivo in vivo Model in vivo imaging involucrin knock-down lead candidate lipid metabolism lipid nanoparticle lipidome lipidomics metabolomics microbial microbicide migration mouse model novel pathogen primary outcome receptor resistance mechanism secondary outcome skin regeneration standard of care treatment effect wound wound biofilm wound closure wound dressing wound healing wound treatment

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项目摘要

ABSTRACT Chronic wounds affect over 2.5 million patients in the US alone, lasting on average 12-13 months and recurring in 60-70% of patients. These occur primarily in lower extremities and vascular disease, diabetes, nephropathy, and metabolic dysfunction are prevalent co-morbidities. These wounds frequently harbor bacterial infections as mixed Gram-negative (e.g. P. aeruginosa, PA) and Gram-positive biofilms (e.g. S. aureus, SA), conferring resistance to antibiotic therapy and inhibiting resolution. The heterogenous biofilm barrier, composed of lipids, proteins, and polysaccharides, presents a robust physiochemical barrier and substrate for persistent infection. Transition of planktonic bacteria to the distinct biofilm state requires a suitable substrate for specific adhesin receptors, such as those found in host membranes and extracellular matrix (ECM), or nonspecific physiochemical binding, enabling aggregation and biofilm ECM organization. This includes changes in microbial lipid composition, secretion of lipid quorum signals and lipase virulence factors, and shift to oxidative lipid metabolism to enable persistent colonization and high-density proliferation. Recent advances in ultra-high- performance liquid chromatography, high resolution mass spectrometry, and bioinformatics technologies have enabled detailed and accurate definition of the bacterial lipidome, aiding in identification of lipid pathways and targets for antimicrobial activity using a systems biology approach. Wound healing is further complicated by pro-inflammatory endogenous phospholipase activities. We propose that lipid ether amines (LEA) represent a platform for novel topical wound treatments with phospholipid anti- metabolite activity, counteracting both host and pathogen pathogenic mechanisms. As preliminary data, we demonstrate >3-log (>1000x) colony forming unit reductions in multiple in vitro cultured, antibiotic resistant biofilms and optimization of anti-metabolite activity. Phase I will synthesize additional candidates, screen for biofilm reduction and activity in host cells, selecting a platform lead to determine in vivo wound healing effects using biofilm infected db/db diabetic mouse model. LC-MS/MS analysis will be used to characterize bacterial or host lipidomes and treatment effect in each Aim. In Aim I, LEA candidate compounds will be screened for antimicrobial effect using clinically relevant, drug resistant ESKAPE (E. faecium, S. aureus, K. pneumoniae, A. baumannii, P. aeruginosa, Enterobacter spp.) pathogens in cultured biofilms, including a translatable wounded ex vivo pig skin explant model. In Aim II, LEA effects on migration and proliferation will be assessed using in vitro full thickness skin construct. In Aim III, the lead compound will be applied in a diabetic mouse wound healing model infected with PA or SA biofilms, to assess in vivo microbicidal and wound closure effects. In Phase II, we will escalate in vivo model complexity by inoculating with mixed bacterial biofilms, refine mechanism using in vitro binding and knock-down/out, evaluate potential adaptive resistance mechanisms, conduct comprehensive ADME-T study, and begin in vivo large mammal Proof of Concept for IND submission.

抽象的慢性伤口仅在美国就会影响超过250万患者，平均持续12-13个月并反复出现在60-70％的患者中。这些主要发生在下肢和血管疾病，糖尿病，肾病，代谢功能障碍是普遍的合并症。这些伤口经常怀有细菌感染混合革兰氏阴性（例如P.铜绿，宾夕法尼亚州）和革兰氏阳性生物膜（例如S. Aureus，sa），授予抵抗抗生素疗法和抑制分辨率。由脂质组成的异质生物膜屏障蛋白质和多糖呈现出强大的生理屏障和底物，以持续感染。浮游细菌向独特的生物膜状态的过渡需要适当的底物来特异性粘附素受体，例如在宿主膜中发现的受体和细胞外基质（ECM）或非特异性的受体生理化学结合，使聚集和生物膜ECM组织。这包括更改微生物脂质组成，脂质群信号和脂肪酶毒力因子的分泌，并转移到氧化脂质代谢可以持续定殖和高密度增殖。超高的最新进展性能液相色谱法，高分辨率质谱和生物信息学技术具有启用了细菌脂肪组的详细和准确的定义，协助识别脂质途径和使用系统生物学方法的抗菌活性的靶标。促炎性内源性磷脂酶活性使伤口愈合更加复杂。我们建议脂质乙醚胺（LEA）代表了新型局部伤口治疗的平台代谢物活性，抵消宿主和病原体致病机制。作为初步数据，我们演示> 3-log（> 1000x）菌落形成单位减少，多种体外培养的抗生素耐药性生物膜和抗代谢物活性的优化。第一阶段将综合其他候选人，屏幕宿主细胞中的生物膜还原和活性，选择平台导致体内伤口愈合效应使用感染生物膜的DB/DB糖尿病小鼠模型。 LC-MS/MS分析将用于表征细菌或每个目标的宿主脂质组和治疗效果。在AIM I中，将筛选LEA候选化合物抗菌作用使用临床相关的耐药性Eskape（E. Faecium，S。Aureus，K。Pneumoniae，A。 Baumannii，铜绿假单胞菌，肠杆菌属。）培养的生物膜中的病原体，包括可翻译的受伤离体猪皮肤外植体模型。在AIM II中，将使用IN评估对迁移和扩散的影响体外厚度全厚的皮肤结构。在AIM III中，铅化合物将应用于糖尿病小鼠伤口感染了PA或SA生物膜的治疗模型，以评估体内微生物和伤口闭合作用。在第二阶段，我们将通过与混合细菌生物膜接种，精炼来升级体内模型的复杂性使用体外结合和敲除/淘汰的机制，评估潜在的自适应抗性机制，进行全面的ADME-T研究，并开始体内大型哺乳动物的概念证明以进行IND提交。