Biologic Evaluation of Liposome-Encapsulated Hemoglobin

脂质体包裹的血红蛋白的生物学评价

• 批准号: 8656751
• 负责人: VIBHUDUTTA AWASTHI
• 金额: $ 37.56万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8656751
• 关键词: Acute; Address; Animal Model; Animals; Antigens; Benign; Biochemical; Biodistribution; Biological Assay; Blood; Blood Circulation; Blood Platelets; Blood Pressure; Brain; Cardiac; Cause of Death; Cerebrum; Cessation of life; Clinical; Colloids; Complement; Complement 3a; Complement 5a; Data; Development; Drug Formulations; Drug Kinetics; Emergency Situation; Encapsulated; Energy Metabolism; Ensure; Enzyme-Linked Immunosorbent Assay; Equipment and supply inventories; Erythrocytes; Evaluation; Evolution; Extravasation; Foundations; Functional disorder; Goals; Half-Life; Heart Rate; Hemoglobin; Hemorrhage; Hemorrhagic Shock; Human; Immune; Immune response; Immunohistochemistry; Infection; Inflammation; Infusion procedures; Injection of therapeutic agent; Injury; Intestines; Investigation; Ischemia; Lactated Ringer' s Solution; Life; Liposomes; Liquid substance; Liver; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Mediating; Metabolism; Microscopy; Military Personnel; Modeling; Monitor; Mus; Nitric Oxide; Operative Surgical Procedures; Organ; Oryctolagus cuniculus; Oxidative Stress; Oxygen; Particle Size; Phospholipids; Physiological; Plasma; Platelet Activation; Population; Positron-Emission Tomography; Preparation; Process; Property; Radiolabeled; Rattus; Reaction; Relative (related person); Reperfusion Injury; Reperfusion Therapy; Research; Respiration; Resuscitation; Reticuloendothelial System; Safety; Secondary to; Serum; Side; Site; Spleen; Sulfur; Technology; Temperature; Testing; Time; Tissues; Toxic effect; Toxin; Transfusion; Trauma; Whole Blood; Work; base; combat; complement C4d; efficacy testing; glucose metabolism; hemodynamics; improved; mortality; neutrophil; novel; optimism; prevent; radiotracer; response; restoration; safety testing; scale up; soluble complement C5b-9; uptake

DESCRIPTION (provided by applicant): Severe hemorrhage is the leading cause of death among trauma victims, and is responsible for approximately 3 million deaths worldwide. Much of the mortality may be prevented by the timely availability of a safe and uni- versal resuscitation fluid. While whole blood or red blood cells (RBCs) remain the preferred transfusion fluids, inadequate availability of safe blood remains a problem. Despite the optimism in combating transfusion-related infections and ensuring an educated donor population, the limited shelf-life of blood and the need to cross- match frequently cause blood inventory shortages. These issues have stimulated a search for safe, immune- tolerant, shelf-stable and efficacious oxygen carrier for both civilian emergency and military applications. He- moglobin based oxygen carriers increase both oxygen-carriage and cardiac preload. We are developing liposome-encapsulated hemoglobin or LEH as a substitute oxygen carrier closely mimick- ing the RBCs. Encapsulated hemoglobin is believed to have better control over hemodynamics as compared to other chemically-modified, but free hemoglobin products. Using a novel formulation and advanced processing technologies, we have overcome the barriers of low encapsulation, oxidative instability, expense and scale-up. The product (called NeoLEH) circulates in blood for a prolonged time, and improves cerebral metabolism. Pre- liminary data demonstrate that the smaller size of LEH (275 nm) as compared to the RBCs (7.5 5m) results in more efficient oxygen delivery to the brain, which in turn improves cerebral oxygen and energy metabolism. In the proposed work, we will test our overall hypothesis that NeoLEH improves survival and cerebral metabol- ism, and serves as a long-circulating and non-toxic oxygen carrier in hemorrhagic shock. The specific aims are to assess: 1) Survival and cerebral metabolism in a rat model of 40% hemorrhage, 2) Pharmacokinetics and biodistribution of NeoLEH, and the effect of reinjection, 3) Acute toxic effects of NeoLEH in the animal model of 40% hemorrhage, and 4) NeoLEH resuscitation and ischemia-reperfusion (I/R) injury. We propose a 4-y project for testing efficacy and safety of NeoLEH. We will study 24 h survival of rats receiv- ing NeoLEH resuscitation after 40% blood loss. Ringer's lactate, shed blood and Hextend will be used as con- trol fluids. Improvement in cerebral oxygen, glucose and energy metabolism will be evaluated by positron emission tomography and magnetic resonance imaging. Circulation persistence and biodistribution of NeoLEH will be assessed by monitoring the blood and organ levels of radiolabeled 99mTc-NeoLEH. In order to test the safety of NeoLEH, acute effects of NeoLEH infusion on phagocytic capacity of liver and complement-mediated platelet activation will be studied. Lastly, we will assess if NeoLEH infusion has any salutary effects in ische- mia-reperfusion phenomenon by monitoring intestinal integrity, neutrophil sequestration, etc. The results of the proposed investigations will be critical for the clinical development of NeoLEH. Targeted towards resuscitation in acute blood loss due to surgery or trauma, NeoLEH will serve civilian as well as military needs alike.

描述（由申请人提供）：严重出血是创伤受害者的主要死亡原因，全球约有300万人死亡。及时提供安全、均匀的复苏液可避免大部分死亡。虽然全血或红细胞（RBC）仍然是首选的输血液体，但安全血液的可用性不足仍然是一个问题。尽管在防治输血相关感染和确保受教育的献血者群体方面持乐观态度，但血液的有限保质期和交叉配血的需要经常导致血液库存短缺。这些问题刺激了对用于民用紧急情况和军事应用的安全、免疫耐受、贮存稳定和有效的氧载体的研究。基于血红蛋白的氧载体增加氧运载和心脏前负荷。我们正在开发脂质体包裹的血红蛋白或莱赫作为替代氧载体，与红细胞非常相似。与其他化学修饰但游离的血红蛋白产品相比，包封的血红蛋白被认为对血液动力学具有更好的控制。使用新的配方和先进的加工技术，我们已经克服了低封装，氧化不稳定性，成本和规模化的障碍。该产品（称为NeoLEH）在血液中循环很长一段时间，并改善大脑代谢。初步数据表明，与RBC（7.5 μ m）相比，较小尺寸的莱赫（275 nm）导致更有效的氧气输送到大脑，这反过来改善了大脑氧气和能量代谢。在本研究中，我们将验证我们的总体假设，即NeoLEH可改善存活率和脑代谢，并在失血性休克中作为长循环和无毒的氧载体。具体目的是评估：1）40%出血大鼠模型中的存活率和脑代谢，2）NeoLEH的药代动力学和生物分布以及再注射的影响，3）NeoLEH在40%出血动物模型中的急性毒性作用，以及4）NeoLEH复苏和缺血-再灌注（I/R）损伤。我们提出了一个为期4年的项目来测试NeoLEH的有效性和安全性。我们将研究40%失血后接受NeoLEH复苏的大鼠的24小时存活率。将使用乳酸林格氏液、脱落血和Hextend作为对照液。将通过正电子发射断层扫描和磁共振成像评价脑氧、葡萄糖和能量代谢的改善。将通过监测放射性标记的99 mTc-NeoLEH的血液和器官水平来评估NeoLEH的循环持久性和生物分布。为了测试NeoLEH的安全性，将研究NeoLEH输注对肝脏吞噬能力和补体介导的血小板活化的急性影响。最后，我们将通过监测肠道完整性、中性粒细胞隔离等来评估NeoLEH输注是否对缺血再灌注现象有任何有益的影响。拟议研究的结果对于NeoLEH的临床开发至关重要。NeoLEH针对因手术或创伤导致的急性失血复苏，将满足民用和军用需求。