Engineering a novel biomaterial for oxygen transport applications

设计用于氧传输应用的新型生物材料

• 批准号: 10545751
• 负责人: Paul Werner Buehler
• 金额: $ 61.39万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-15 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10545751
• 关键词: Animal Model; Animals; Antioxidants; Arteriosclerosis; Biocompatible Materials; Biotechnology; Blood; Blood Preservation; Blood Vessels; Blood Volume; Cardiac; Cattle; Cavia; Chemicals; Circulation; Clinical; Clinical Trials; Complex; Complex Mixtures; Development; Diameter; Diet; Dose; Drug Kinetics; Electron Spin Resonance Spectroscopy; Emerging Communicable Diseases; Endothelium; Engineering; Enzymes; Erythrocytes; Event; Exhibits; Extravasation; Fatty acid glycerol esters; Generations; Glutaral; Goals; Hemoglobin; Hemorrhage; Heterogeneity; High Fat Diet; Human; Image; Impairment; Individual; Injury; Laboratories; Light; Measures; Membrane; Methods; Modeling; Modification; Molecular; Molecular Weight; Myocardial Infarction; Nanostructures; Nitric Oxide; Oxidative Stress Induction; Oxygen; Oxygen saturation measurement; Phase III Clinical Trials; Physiological; Plasma; Polymers; Population; Proteins; RNA Splicing; Recombinants; Regulation; Reporting; Research; Reticuloendothelial System; Risk; Role; Sampling; Signal Transduction; Structure; Sucrose; Systemic hypertension; Testing; Therapeutic; Tissues; Toxic effect; Trans-Splicing; Transfusion; Translating; Ultrafiltration; Vascular Permeabilities; Vascular blood supply; Whole Blood Exchange Transfusion; Work; biophysical techniques; design; design and construction; dimer; endothelial dysfunction; femoral artery; hemoglobin polymer; hypertensive; innovation; intein; manufacture; molecular size; monomer; mutant; nonalcoholic steatohepatitis; novel; oxidative damage; oxygen transport; physical property; preclinical trial; primary endpoint; repaired; response; screening; side effect; survival outcome; tissue injury; tissue oxygenation; transfusion medicine; vascular injury; vasoconstriction

Abstract Hemoglobin (Hb)-based oxygen (O2) carriers (HBOCs) are currently being developed as red blood cell (RBC) substitutes for use in transfusion medicine. Despite significant commercial development, recent late stage clinical results of polymerized hemoglobin (PolyHb) solutions (i.e. Hemopure (OPK Biotech, Cambridge, MA), a glutaraldehyde polymerized bovine Hb; and PolyHeme (Northfield Laboratories Inc., Evanston, IL), a glutaraldehyde polymerized pyridoxylated human Hb) hamper further development. Both of these commercial products elicit vasoconstriction at the microcirculatory level, and lead to the development of systemic hypertension and oxidative tissue damage. These side-effects are hypothesized to occur either by a nitric oxide (NO) scavenging or oxygen (O2) oversupply mechanism, and are both exacerbated by PolyHb extravasation into the tissue space. In light of these 2 potential mechanisms, it is apparent that PolyHb size will have a profound impact on the extent of vasoconstriction, systemic hypertension and oxidative tissue toxicity. However, commercial PolyHb products are complex mixtures with broad size distributions defined only by the size cutoff of the ultrafiltration membranes used in their manufacture. Furthermore, these mixtures are known to contain up to 1% of individual tetrameric Hb molecules and a significantly higher proportion of lower molecular weight (MW) Hb oligomers (80% with MW < 500 kDa). Hence, the side-effects observed during clinical/pre-clinical trials are attributed to a mixture of low MW Hb polymers with different sizes and points of chemical modification, and not to any one, single PolyHb molecule. This precludes precise characterization of how individual components of these complex PolyHb mixtures interact with the vasculature. An important advance would therefore be the ability to produce molecularly uniform, monodisperse, and high MW PolyHb nanostructures. In this application, we hypothesize that the molecular diameter and topology of recombinant PolyHb (rPolyHb) will regulate vasoactivity and oxidative injury to tissues. To test our hypothesis we propose the following specific aims: Specific Aim 1: Use orthogonal split splicing inteins to produce well-defined, monodisperse, high MW rPolyHb nanostructures. Specific Aim 2a: Analyze the role of endothelial function on the development of vasoactivity and oxidative tissue injury to rPolyHbs of varying size. Specific Aim 2b: Evaluate the pharmacokinetics of rPolyHbs in normal guinea pigs and HFSD guinea pigs. Specific Aim 3: Evaluate the ability of rPolyHbs to restore tissue oxygenation and optimize survival in severe blood loss. The proposed work is both significant and innovative, since it seeks to develop safe and efficacious rPolyHbs for use in transfusion medicine. In addition, state-of-the-art biophysical techniques and two unique animal models will be used to understand rPolyHb physiological responses and determine the clinical potential of these novel materials.

抽象的 基于血红蛋白 (Hb) 的氧 (O2) 载体 (HBOC) 目前正在开发为红细胞 （红细胞）替代品用于输血医学尽管商业发展显着，但最近进展较晚。 聚合血红蛋白 (PolyHb) 溶液（即 Hemopure（OPK Biotech， Cambridge, MA)，一种戊二醛聚合牛 Hb；和 PolyHeme（Northfield Laboratories Inc.， 伊利诺伊州埃文斯顿），一种戊二醛聚合吡啶氧化人血红蛋白）阻碍了两者的进一步发展。 这些商业产品引起微循环水平的血管收缩，并导致开发 全身性高血压和氧化组织损伤可能会因以下原因再次出现。 一氧化氮 (NO) 清除或氧气 (O2) 供应过剩机制，并且都会因 PolyHb 而加剧 鉴于这两种潜在机制，PolyHb 的大小显然会外渗到组织间隙中。 对血管收缩、全身性高血压和氧化组织毒性的程度有深远的影响。 然而，商业 PolyHb 产品是复杂的混合物，具有广泛的尺寸分布，仅由 此外，这些混合物的制造中使用的超滤膜的尺寸截留。 已知含有高达 1% 的单个四聚 Hb 分子，并且较低比例的 Hb 分子比例明显更高 分子量 (MW) Hb 寡聚物（MW < 500 kDa 时为 80%） 因此，在过程中观察到了副作用。 临床/临床前试验归因于具有不同尺寸和点的低MW Hb聚合物的混合物 化学修饰，而不是对任何一个单一的 PolyHb 分子，这妨碍了对 PolyHb 的精确表征。 这些复杂的 PolyHb 混合物的各个成分如何与脉管系统相互作用。 因此，一个重要的进步是能够生产分子均匀、单分散和 高分子量多聚血红蛋白纳米结构在此应用中，我们追求分子直径和 重组多聚血红蛋白 (rPolyHb) 的拓扑结构将调节血管活性和组织氧化损伤。 检验我们的假设，我们提出以下具体目标： 具体目标 1：使用正交分裂剪接内含子产生明确的、单分散的、高分子量的 rPolyHb 纳米结构。 具体目标 2a：分析内皮功能对血管活性和氧化发展的作用 不同大小的 rPolyHbs 造成的组织损伤。 具体目标 2b：评估 rPolyHbs 在正常豚鼠和 HFSD 豚鼠中的药代动力学 猪。 具体目标 3：评估 rPolyHbs 恢复组织氧合和优化生存的能力 严重失血。 拟议的工作既重要又具有创新性，因为它旨在开发安全有效的药物 rPolyHbs 用于输血医学此外，最先进的生物物理技术和两种独特的技术。 动物模型将用于了解 rPolyHb 生理反应并确定临床潜力 这些新颖的材料。