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，特别是当并发低血压等继发性损伤时。TBI患者对低血压的易感性是公认的，并确定了新方法的关键需求。本提案的目标是开发一种神经保护、超胶体、氧治疗、多硝基化聚乙二醇化血红蛋白（PNPH），用于TBI合并低血压的院前复苏。在Safar复苏研究中心和SynZyme Technologies（由美国陆军资助）为期4年的合作中，先前基于使用具有共价连接抗氧化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PNPH这种药物具有令人兴奋的潜力。PNPH有三个组成部分有助于其独特的治疗活性：1)作为蛋白质中心的血红蛋白提供氧气输送能力；2)PNPH的聚羟基化部分提供超胶体特性，对低血压期间稳定血流动力学至关重要；3)PNPH的氮氧化物部分不仅提高无细胞血红蛋白的安全性，还提供抗氧化/抗炎和神经保护活性。几十年来，人们的注意力仅仅集中在血红蛋白携带氧气的能力上。在创伤性脑损伤的情况下，治疗不仅要提供氧气，还要保护神经元。多硝基化将促氧化血红蛋白转化为神经元保护氧载体，这是提出的治疗的理想候选者。本文介绍了用羧基（CO）牛聚乙二醇化血红蛋白多硝基化制备PNPH的初步研究。体外研究表明，PNPH是一种独特的血红蛋白，具有神经保护而非神经毒性。同样，体内研究表明，该PNPH作为小体积复苏溶液，在小鼠TBI + HS后，可最大限度地提高血流动力学稳定性、存活率和脑组织氧水平，同时最大限度地减少神经元死亡，优于目前民用和军用院前TBI复苏的护理标准。在这个转化项目的第一阶段，将对质量控制的PNPH进行评估，以确定产品规格、功能结果的有效性和初步毒理学。这将为ind前会议获得FDA指导奠定基础。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毒理学和安全性测试，为在拨款期结束时提交IND做准备。

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.