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）：该多学科生物工程研究伙伴关系（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、表面活性和肺疗效。还研究了使用合成表面活性剂促进外源DNA向肺损伤动物的传递，以用于未来的基因治疗或多药治疗。BRP的主要目标是开发与现有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、纯度和生产经济性的全合成肺表面活性剂，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）。一个在生物工程、物理、化学、生物学和医学方面经验丰富的协作式BRP团队聚集在三所大学：罗切斯特大学（主要机构）、洛杉矶生物医学研究所/港湾-加州大学洛杉矶分校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关键分子特征的化合物，SP -B在天然表面活性剂中具有重要的功能活性。与人SP-C/SP-A相关的多肽和脂肽也被研究，并通过生物工程使其相对于天然载脂蛋白具有分子稳定性优势。BRP还检测了两种类型的脂质：模仿天然表面活性剂的合成脂质(L)，以及在炎症性肺损伤（ALI/ARDS）中具有增强吸附和扩散以及抵抗降解能力的新型磷脂酶抗性脂质（RL）。目标1和目标2研究生物工程、蛋白质组学、合成、纯化、分子生物物理学以及基于有希望的初始活性化合物的合成肽/脂肽和新型RL化合物的规模化。目的3研究基于体外表面活性评估（脉动泡、吸附、威廉平衡、捕获泡）和动物肺活性研究的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认可的切除洗肺大鼠肺力学模型，用于评估目前临床表面活性剂药物直接用于NRDS早产儿；（ii）气管内注射脂多糖（LPS）对体内ALI/ARDS小鼠的影响；(3)体内严重高氧暴露致ALI/ARDS家兔；(4)表面活性剂缺乏和体内灌洗引起的ALI/ARDS通气家兔。目的4研究了灌注合成表面活性剂的剪切粘度、肺分布和细胞毒性，以及它们在促进DNA肺递送到lps小鼠中的应用，为未来ALI/ARDS的基因或多药治疗提供帮助。这项多学科BRP资助将加强对多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，以生物工程的方式合成具有最大活性、抗抑制性、肺疗效和生产经济性的肺表面活性剂。公共卫生相关性：这项BRP研究将开发和生产具有最大活性和抑制抗性的新型全合成肺表面活性剂，用于未来治疗包括急性呼吸衰竭在内的严重和流行的人类肺部疾病。BRP表面活性剂将经过生物工程改造，比目前合成的表面活性剂药物具有更强的活性和抗抑制能力，并且与现有的动物源性临床表面活性剂相比，在活性、耐药性、纯度、可重复性、稳定性和生产经济性方面也具有显著的潜在优势。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影响数十万各个年龄段（婴儿到成人）的患者。通过检测合成表面活性剂在改善呼吸衰竭方面的主要活性，以及它们促进肺内DNA传递的能力，这些研究不仅与公共卫生有进一步的关联，还有助于未来基因治疗或多药物治疗方法治疗ALI/ARDS和其他肺部疾病。