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）：这种多学科生物工程研究伙伴关系（BRP）授予研究了新型全合成脂质/肽肺表面活性剂的分子生物工程，合成，表面活性和肺功能。还研究的是使用合成表面活性剂促进外源性DNA向肺损伤的动物递送，以进行未来的基因治疗或多药治疗应用。 BRP的主要目的是与现有动物衍生或合成的临床表面活性药物相比，开发具有最大活性，抑制性，稳定性，纯度和生产经济性的全合成肺表面活性剂，用于治疗新生儿呼吸呼吸疾病综合征（NRDS），急性肺部呼吸损伤（ALI）和急性急性急性（ARD）（急性呼吸道）。一支具有丰富经验的合作BRP团队，在生物工程，物理，化学，生物学和医学方面具有专业知识，在三所大学中组装：罗切斯特大学（小学机构），LA BioMedical Research Institute/Harbour-UCLA医学中心和圭尔夫大学。在BRP中研究的合成肽包括包含人表面活性剂蛋白（SP）-b的关键分子特征的化合物，该蛋白（SP）-B具有至关重要的功能活性。还研究了与人类SP-C/SP-A相关的肽和脂肽，并进行生物工程具有相对于天然顶蛋白具有分子稳定性的优势。 BRP还检查了两种类型的脂质：以天然表面活性剂为模型的合成脂质（L），以及具有增强的吸附和扩散的新型磷脂酶耐药性脂质（RL），以及抵抗炎症性肺损伤（ALI/ARDS）的降解能力。目标1和2研究了基于在初始活性化合物的有希望的初步数据，研究了合成肽/脂蛋白/脂蛋白肽/脂蛋白肽和新型RL化合物的生物工程，蛋白质组学，合成，纯化，分子生物物理学。 AIM 3研究了基于体外表面活性评估（脉动气泡，吸附，Wilhelmy平衡，圈养气泡）和动物中的肺活性研究，研究了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）一种被切除的灌输大鼠肺机械模型，该模型被FDA被接受，用于评估当前的临床表面活性药物，以直接用于NRDS的早产婴儿； （ii）从气管内滴注脂多糖（LPS）的体内ali/ards的小鼠； （3）因严重高氧化暴露而在体内具有ali/ards的兔子； （4）体内灌洗引起的表面活性剂缺陷和ALD的通风兔子。 AIM 4研究了所灌输的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递送到LPS-小鼠的肺部的实用性，用于将来的应用，涉及涉及ALI/ARDS的Gene或Multi-drug疗法。这种多学科的BRP赠款将增强对肽和脂质的分子行为的科学理解，同时使用具有最大活性，抑制耐药性，肺部效率和生产经济性的最大活性，抑制活性，抑制活性，抑制活性的原理，化学，物理学，生物学，生理和医学。公共卫生相关性：这项BRP研究将开发并产生具有最大活性和抑制性的新型全合成肺表面活性剂，以在治疗涉及急性呼吸衰竭的严重和普遍的人类肺部疾病中使用。与当前的合成表面活性剂药物相比，BRP表面活性剂将被生物工程更具活性和抑制作用，并且与现有动物衍生的临床表面活性剂相比，在活动，耐药性，纯度，可重复性，稳定性，稳定性，稳定性和生产经济方面也将具有显着的潜在优势。特定的治疗应用不仅包括早产婴儿中的新生儿呼吸窘迫综合征（NRDS），还包括临床急性肺损伤（ALI）和急性呼吸遇险综合征（ARDS），这些疾病（ARDS）每年都会影响美国和世界各地的所有年龄段（婴儿）的数十万患者。补助金的增加研究与公共卫生有进一步的相关性，不仅是为了改善呼吸衰竭的主要活性，而且还可以促进DNA的肺部递送DNA用于未来的基因治疗或多药治疗方法，以治疗ALI/ARD和其他肺部疾病。