Polynitroxylated Pegylated Hemoglobin for Traumatic Brian Injury Resuscitation

描述（由申请人提供）：创伤性脑损伤（TBI）是发病率和死亡率的主要原因，尤其是在并发二次损伤（如低血压）时。创伤性脑损伤患者易发生低血压已得到充分认识，并确定了新方法的关键需求。本提案的目的是开发一种神经保护性、高胶体、氧治疗性、多硝基聚乙二醇化血红蛋白（PNPH），用于TBI合并低血压的院前复苏。在美国陆军资助的Safar复苏研究中心和SynZyme Technologies之间为期4年的合作中，先前基于使用具有共价连接的抗氧化剂氮氧化物部分的蛋白质的策略筛选了几种解决方案，产生了初步数据，表明这种药剂PNPH具有令人兴奋的潜力。PNPH具有三种组分，有助于其独特的治疗活性：1）血红蛋白作为蛋白质中心提供氧输送能力; 2）PNPH的聚乙二醇化部分提供对稳定低血压期间的血流动力学很重要的超胶体性质; 3）PNPH的氮氧部分不仅改善无细胞血红蛋白的安全性，而且还提供抗氧化/抗炎和神经保护活性。几十年来，人们的注意力一直集中在血红蛋白携带氧气的能力上。在TBI的情况下，治疗不仅要提供氧气，还要保护神经元。多硝酰化将促氧化血红蛋白转化为神经元保护氧载体，这是所提出的治疗的理想候选物。PNPH的初步研究，制备的羧基（CO）牛聚乙二醇化血红蛋白的多硝酰化。体外研究表明，PNPH是一种独特的血红蛋白，具有神经保护作用而非神经毒性。类似地，体内研究表明，这种PNPH用作小体积复苏溶液，其在小鼠中TBI加HS后最大化血液动力学稳定性、存活率和脑组织氧水平，同时最小化神经元死亡，并且优于民用和军用院前TBI复苏的当前护理标准。在该转化项目的I期，将对质量受控的PNPH进行评价，以确定产品质量标准、功能结局疗效和初步毒理学。这将为IND前会议奠定基础，以获得FDA的指导。在第二阶段，将扩大生产规模，并进行必要的CMC和FDA要求的GLP毒理学和安全性试验，为在授权期结束时提交IND做准备。 公共卫生相关性：创伤性脑损伤（TBI）是民用和军事环境中发病率和死亡率的主要原因。二次损伤，如多发性创伤伴出血性休克（HS）是常见的，特别是在严重TBI的情况下，并对结局产生破坏性影响。在当前的护理标准下，脑损伤患者对低血压和脑水肿的脆弱性是公认的，并确定了对新方法的关键需求。本申请提出将治疗性多硝基化聚乙二醇化血红蛋白（PNPH）纳入IND，PNPH已被证明是一种独特的神经保护性血红蛋白，其用作小体积复苏溶液，在小鼠TBI加HS后最大化血液动力学稳定性、存活率和脑组织氧水平，同时最小化神经元死亡，并且优于民用和军用院前TBI复苏的当前护理标准。

DESCRIPTION (provided by applicant): Traumatic brain injury (TBI) is a leading cause of morbidity and mortality especially when complicated by secondary insults such as hypotension. Vulnerability of TBI patients to hypotension is well recognized and identifies a key need for new approaches. The goal of this proposal is to develop to IND a neuroprotective, hyper-colloid, oxygen therapeutic, polynitroxylated pegylated hemoglobin (PNPH), for pre-hospital resuscitation in the setting of TBI complicated by hypotension. In a 4-year collaboration between Safar Center for Resuscitation Research and SynZyme Technologies funded by the US Army, previous screening of several solutions based on the strategy of using proteins with covalently linked antioxidant nitroxide moieties produced preliminary data suggesting exciting potential for one such agent, PNPH. PNPH has three components contributing to its unique therapeutic activities: 1) hemoglobin as the protein center provides oxygen delivery capabilities 2) the pegylation moieties of PNPH provide hyper-colloid properties important to stabilizing hemodynamics during hypotension, and 3) the nitroxide moieties of PNPH not only improve the safety of cell-free hemoglobin but also provide anti-oxidant/anti-inflammatory and neuroprotective activities. For decades, attention has focused solely on hemoglobin's ability to carry oxygen. In the case of TBI, it is essential that the treatment not only delivers oxygen but also protects neurons. Polynitroxylation converts pro-oxidant hemoglobin into a neuron-protective oxygen carrier, which is an ideal candidate for the proposed therapy. Preliminary studies of PNPH, prepared by polynitroxylation of carboxy (CO) bovine pegylated hemoglobin are presented. In vitro studies suggest that PNPH is a unique hemoglobin with neuroprotective rather than neurotoxic properties. Similarly, in vivo studies show that this PNPH functioned as a small volume resuscitation solution that maximized hemodynamic stability, survival and brain tissue oxygen levels while minimizing neuronal death, after TBI plus HS in mice and outperformed the current standard of care for civilian and military pre-hospital TBI resuscitation. In the phase I of this translational program, quality controlled PNPH will be evaluated to determine product specifications, efficacy in functional outcome and preliminary toxicology. This will set the stage for a pre-IND meeting to obtain FDA guidance. In phase II, production will be scaled up and requisite CMC and FDA required GLP toxicology and safety testing will be performed in preparation for IND submission at the end of the grant period. PUBLIC HEALTH RELEVANCE: Traumatic brain injury (TBI) is a leading cause of morbidity and mortality in both civilian and military settings. Secondary insults, such as polytrauma with hemorrhagic shock (HS) are common, especially in cases of severe TBI, and have a devastating impact on outcome. Vulnerability of brain-injured patients to hypotension and brain edema with the current standard of care is well recognized and identifies a key need for new approaches. This application proposes to bring to IND a therapeutic, polynitroxylated pegylated hemoglobin (PNPH), which has been shown to be a unique neuroprotective hemoglobin that functioned as a small volume resuscitation solution that maximized hemodynamic stability, survival and brain tissue oxygen levels while minimizing neuronal death, after TBI plus HS in mice and outperformed the current standard of care for civilian and military pre-hospital TBI resuscitation.