Polynitroxylated Pegylated Hemoglobin for Traumatic Brian Injury Resuscitation

描述(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，特别是当合并二次损伤时，如低血压。脑外伤患者对低血压的易感性得到了很好的认识，并确定了新方法的关键需求。这项建议的目标是开发一种神经保护性、高胶体、氧疗、多氮氧基化聚乙二醇化血红蛋白(PNPH)，用于颅脑损伤合并低血压的院前复苏。在由美国陆军资助的萨法尔复苏研究中心和SynZyme Technologies之间为期4年的合作中，之前基于使用带有共价连接的抗氧化剂氮氧化物部分的蛋白质的策略对几种解决方案进行的筛选产生了初步数据，表明这种制剂PNPH具有令人兴奋的潜力。PNPH具有三种独特的治疗活性：1)作为蛋白质中心的血红蛋白提供氧气输送能力；2)PNPH的聚乙二醇化部分提供对低血压期间稳定血流动力学至关重要的高胶体性质；3)PNPH的氮氧化物部分不仅提高了无细胞血红蛋白的安全性，还具有抗氧化/抗炎和神经保护活性。几十年来，人们的注意力只集中在血红蛋白携带氧气的能力上。在脑外伤的情况下，至关重要的是治疗不仅要输送氧气，还要保护神经元。多氮氧基化将促氧化血红蛋白转化为神经元保护的氧载体，这是建议的治疗方案的理想候选者。本文报道了由羧基(CO)聚乙二醇化牛血红蛋白多氮氧基化制备PNPH的初步研究。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，而不是神经毒性。同样，体内研究表明，这种PNPH可以作为小容量复苏液发挥作用，在小鼠脑创伤加HS后最大限度地提高血流动力学稳定性、存活率和脑组织含氧量，同时最大限度地减少神经元死亡，并且优于目前民用和军用脑创伤院前复苏的护理标准。在该翻译计划的第一阶段，将对PNPH进行质量控制评估，以确定产品规格、功能结果的疗效和初步毒理学。这将为IND前会议获得FDA的指导奠定基础。在第二阶段，生产规模将扩大，必要的CMC和FDA要求的GLP毒理和安全测试将进行，为在授权期结束时提交IND做准备。 公共卫生相关性：在平民和军事环境中，创伤性脑损伤(TBI)都是导致发病率和死亡率的主要原因。继发性损伤，如多发伤合并失血性休克(HS)是很常见的，尤其是在严重的颅脑损伤病例中，对预后有毁灭性的影响。在目前的护理标准下，脑损伤患者对低血压和脑水肿的脆弱性得到了很好的认识，并确定了新方法的关键需要。这项申请旨在为IND带来一种治疗性的多氮氧基化聚乙二醇化血红蛋白(PNPH)，它已被证明是一种独特的神经保护性血红蛋白，可作为小容量复苏液发挥作用，在小鼠脑外伤加HS后最大化血流动力学稳定性、存活率和脑组织氧水平，同时将神经元死亡降至最低，并且优于目前民用和军用脑外伤院前复苏的护理标准。

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.