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 目前正在发展为红细胞的血红蛋白（HB）氧气（O2）载体（HBOC） （RBC）用于输血医学的替代品。尽管商业开发很大，但最近很晚 聚合血红蛋白（PolyHB）溶液的阶段临床结果（即血液蛋白酶（OPK Biotech， 马萨诸塞州剑桥），一种聚合的谷氨酸牛HB；和Polyheme（Northfield Laboratories Inc.， Evanston，伊利诺伊州，一种戊二醛聚合的吡ido氧化毒素HB）阻碍进一步发展。两个 这些商业产品在微循环水平上引起血管收缩，并导致 全身性高血压和氧化组织损伤。这些副作用被认为是由 一氧化氮（NO）清除或氧气（O2）过度供应机制，并且都被Polyhb加剧了 渗入组织空间。鉴于这两种潜在机制，很明显Polyhb的大小将 对血管收缩，全身性高血压和氧化组织毒性的程度产生深远影响。 但是，商业polyhb产品是复杂的混合物，其尺寸分布仅由 其制造中使用的超滤机制的尺寸截止。此外，这些混合物是 已知最多包含1％的单个四聚体HB分子，较低的比例明显更高 分子量（MW）HB低聚物（80％，MW <500 kDa）。因此，在 临床/临床前试验归因于低MW HB聚合物的混合物，大小不同 化学修饰，而不是任何一个单个polyHb分子。这排除了精确表征 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，重要的进步将是产生分子均匀，单分散和 高MW PolyHB纳米结构。在此应用中，我们假设分子直径和 重组PolyHB（RPOLYHB）的拓扑结构将调节组织的血管活性和氧化性损伤。到 检验我们的假设我们提出以下特定目的： 特定目标1：使用正交拆分整数生成定义明确的单分散，高MW RPOLYHB纳米结构。 特定目的2a：分析内皮功能在血管活性和氧化发展中的作用 rpolyhbs的组织损伤不同。 特定目标2B：评估普通豚鼠和HFSD几内亚的Rpolyhbs的药代动力学 猪。 特定目的3：评估RpolyHbs恢复组织氧合并优化生存的能力 严重失血。 拟议的工作既重要又创新，因为它试图发展安全有效 用于输血医学的rpolyhbs。此外，最先进的生物物理技术和两种独特 动物模型将用于了解RPOLYHB的身体反应并确定临床潜力 这些新型材料。