Bioengineering of Novel Synthetic Lipid-Peptide Lung Surfactants

新型合成脂肽肺表面活性剂的生物工程

• 批准号: 8304346
• 负责人: ROBERT H NOTTER
• 金额: $ 59.84万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8304346
• 关键词: Academic Medical Centers; Acute; Acute Lung Injury; Acute respiratory failure; Adsorption; Adult; Adult Respiratory Distress Syndrome; Affect; Age; Amino Acid Substitution; Amino Acids; Animal Model; Animals; Apoproteins; Behavior; Binding; Bioinformatics; Biological Assay; Biology; Biomedical Engineering; Biomedical Research; Biophysics; C-Peptide; Calcium ion; Cell Line; Charge; Chemical Surfactants; Chemistry; Clinical; DNA; DNA delivery; Data; Disulfides; Drug Formulations; Engineering; Equilibrium; Erythrocytes; Esters; Exhibits; FDA approved; Family; Fluorescence; Future; Genes; Glycerophospholipids; Goals; Grant; Guidelines; Health; High Pressure Liquid Chromatography; Human; In Situ; In Vitro; Infant; Infasurf; Inflammatory; Institution; Irrigation; Label; Length; Ligation; Link; Lipids; Lipopolysaccharides; Liquid substance; Lung; Lung diseases; Measures; Mechanics; Medicine; Methods; Modeling; Molecular; Mus; NMR Spectroscopy; National Heart, Lung, and Blood Institute; Neck; Newborn Respiratory Distress Syndrome; Oryctolagus cuniculus; Palmitates; Patients; Peptide Synthesis; Peptides; Pharmaceutical Preparations; Pharmacotherapy; Phosphatidyl glycerol; Phosphatidylglycerols; Phospholipase; Physics; Physiological; Physiology; Premature Infant; Preparation; Production; Proteins; Proteomics; Public Health; Pulmonary Surfactant-Associated Protein A; Pulmonary Surfactant-Associated Protein B; Pulmonary Surfactant-Associated Protein C; Pulmonary Surfactants; Quantitative Microscopy; Rattus; Relative (related person); Reproducibility; Research; Research Institute; Resistance; Respiration; Respiratory Failure; Sonication; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Spectroscopy, Fourier Transform Infrared; Structure; Surface; Survanta; Testing; Therapeutic; Type II Epithelial Receptor Cell; United States; Universities; Viscosity; amyloid formation; analog; arginyllysine; base; clinically significant; cytotoxicity; design; dimer; experience; gene therapy; improved; in vitro activity; in vivo; inhibitor/antagonist; interfacial; lung injury; microwave electromagnetic radiation; multidisciplinary; novel; peptide B; peptide L; research clinical testing; scale up; simulation; solid state; surfactant; surfactant deficiency; synthetic peptide; thioether

DESCRIPTION (provided by applicant): This multidisciplinary bioengineering research partnership (BRP) grant studies the molecular bioengineering, synthesis, surface activity, and pulmonary efficacy of novel fully-synthetic lipid/peptide lung surfactants. Also studied is the use of synthetic surfactants to facilitate the delivery of exogenous DNA to animals with lung injury for future gene therapy or multi-drug therapy applications. The primary goal of the BRP is to develop fully-synthetic lung surfactants with maximal activity, inhibition resistance, stability, purity, and production economy compared to existing animal-derived or synthetic clinical surfactant drugs for treating the neonatal respiratory distress syndrome (NRDS), acute lung injury (ALI), and the acute respiratory distress syndrome (ARDS). A highly-experienced, collaborative BRP team with expertise in bioengineering, physics, chemistry, biology, and medicine is assembled at three universities: the University of Rochester (primary institution), LA Biomedical Research Institute/Harbor-UCLA Medical Center, and the University of Guelph. Synthetic peptides studied in the BRP include compounds incorporating key molecular features of human surfactant protein (SP)-B, which has crucial functional activity in native surfactant. Peptides and lipopeptides related to human SP-C/SP-A are also studied and are bioengineered to have advantages in molecular stability relative to native apoproteins. The BRP also examines two types of lipids: synthetic lipids (L) modeled after those in native surfactant, and novel phospholipase-resistant lipids (RL) having enhanced adsorption and spreading plus the ability to resist degradation in inflammatory lung injury (ALI/ARDS). Aims 1 and 2 study the bioengineering, proteomics, synthesis, purification, molecular biophysics, and scale-up of synthetic peptides/lipopeptides and novel RL compounds based on promising preliminary data on initial active compounds. Aim 3 investigates the optimization of lipid/peptide composition in synthetic surfactants based on surface activity assessments in vitro (pulsating bubble, adsorption, Wilhelmy balance, captive bubble) and pulmonary activity studies in animals. Animal models studied include: (i) an excised lavaged rat lung mechanical model that is FDA-accepted for evaluating current clinical surfactant drugs for direct use in premature infants with NRDS; (ii) mice with ALI/ARDS in vivo from intratracheal instillation of lipopolysaccharide (LPS); (3) rabbits with ALI/ARDS in vivo from severe hyperoxic-exposure; and (4) ventilated rabbits with surfactant-deficiency and ALI/ARDS induced by in vivo lavage. Aim 4 studies the shear viscosity, pulmonary distribution, and cytotoxicity of instilled synthetic surfactants, as well as their utility in facilitating the pulmonary delivery of DNA to LPS-mice for future applications involving gene- or multi-drug therapies for ALI/ARDS. This multidisciplinary BRP grant will enhance scientific understanding about the molecular behavior of peptides and lipids while using principles and methods of engineering, chemistry, physics, biology, physiology and medicine to bioengineer synthetic lung surfactants having maximum activity, inhibition resistance, pulmonary efficacy, and production economy. PUBLIC HEALTH RELEVANCE: This BRP research will develop and produce novel fully-synthetic lung surfactants having maximum activity and inhibition resistance for future use in treating severe and prevalent human pulmonary diseases involving acute respiratory failure. BRP surfactants will be bioengineered to be more active and inhibition-resistant than current synthetic surfactant drugs, and will also have significant potential advantages in activity, resistance, purity, reproducibility, stability, and production economy compared to existing animal-derived clinical surfactants. Specific therapeutic applications include not only the neonatal respiratory distress syndrome (NRDS) in premature infants, but also clinical acute lung injury (ALI) and the acute respiratory distress syndrome (ARDS) that affect hundreds of thousands of patients of all ages (infants to adults) each year in the United States and around the world. Added studies in the grant have further relevance to public health by examining synthetic surfactants not only for their primary activity in improving respiratory failure, but also for their ability to facilitate the pulmonary delivery of DNA for future gene therapy or multi-drug therapy approaches for treating ALI/ARDS and other lung diseases.

描述（由申请人提供）：这项多学科生物工程研究合作伙伴关系（BRP）资助研究新型全合成脂质/肽肺表面活性剂的分子生物工程、合成、表面活性和肺功效。还研究了使用合成表面活性剂来促进将外源 DNA 递送至肺损伤的动物，以用于未来的基因治疗或多药治疗应用。 BRP的主要目标是开发全合成肺表面活性剂，与现有的动物源性或合成临床表面活性剂药物相比，具有最大活性、抗抑制性、稳定性、纯度和生产经济性，用于治疗新生儿呼吸窘迫综合征（NRDS），急性肺损伤（ALI）和急性呼吸窘迫综合征（ARDS）。一支经验丰富的协作 BRP 团队在生物工程、物理、化学、生物学和医学领域拥有专业知识，由三所大学组成：罗切斯特大学（主要机构）、洛杉矶生物医学研究所/港口-加州大学洛杉矶分校医疗中心和大学圭尔夫。 BRP 研究的合成肽包括结合了人表面活性剂蛋白 (SP)-B 关键分子特征的化合物，该蛋白在天然表面活性剂中具有重要的功能活性。还研究了与人SP-C/SP-A相关的肽和脂肽，并对其进行了生物工程改造，使其相对于天然脱辅基蛋白在分子稳定性方面具有优势。 BRP 还检查两种类型的脂质：以天然表面活性剂为模型的合成脂质 (L)，以及具有增强吸附和扩散以及抵抗炎症性肺损伤 (ALI/ARDS) 降解的能力的新型抗磷脂酶脂质 (RL) 。目标 1 和 2 基于初始活性化合物有希望的初步数据，研究生物工程、蛋白质组学、合成、纯化、分子生物物理学以及合成肽/脂肽和新型 RL 化合物的放大。目标 3 基于体外表面活性评估（脉动气泡、吸附、威廉平衡、圈养气泡）和动物肺活性研究，研究合成表面活性剂中脂质/肽组成的优化。研究的动物模型包括：(i) FDA 接受的离体灌洗大鼠肺机械模型，用于评估当前临床表面活性剂药物是否直接用于患有 NRDS 的早产儿； (ii)因气管内滴注脂多糖(LPS)而体内患有ALI/ARDS的小鼠； (3) 因严重高氧暴露而体内患有ALI/ARDS的兔子； (4) 表面活性剂缺乏和体内灌洗诱发 ALI/ARDS 的通气兔。目标 4 研究滴注合成表面活性剂的剪切粘度、肺部分布和细胞毒性，以及它们在促进 DNA 向 LPS 小鼠肺部输送方面的效用，以用于未来涉及 ALI/ARDS 基因或多药治疗的应用。这项多学科 BRP 资助将增强对肽和脂质分子行为的科学理解，同时利用工程、化学、物理、生物学、生理学和医学的原理和方法来生物工程合成肺表面活性剂，使其具有最大的活性、抑制阻力、肺功效和产量经济。公共健康相关性：这项 BRP 研究将开发和生产具有最大活性和抗抑制性的新型全合成肺表面活性剂，用于未来用于治疗涉及急性呼吸衰竭的严重和普遍的人类肺部疾病。 BRP表面活性剂将经过生物工程改造，比现有的合成表面活性剂药物具有更高的活性和抗抑制性，并且与现有的动物源临床表面活性剂相比，在活性、耐药性、纯度、重现性、稳定性和生产经济性方面也将具有显着的潜在优势。具体的治疗应用不仅包括早产儿的新生儿呼吸窘迫综合征 (NRDS)，还包括影响数十万各个年龄段（婴儿到成人）患者的临床急性肺损伤 (ALI) 和急性呼吸窘迫综合征 (ARDS) ）每年在美国和世界各地。拨款中增加的研究与公共健康进一步相关，通过检查合成表面活性剂，不仅检查其改善呼吸衰竭的主要活性，还检查其促进 DNA 肺部输送的能力，以用于未来的基因治疗或多药治疗方法。 ALI/ARDS 和其他肺部疾病。