rhMFG-E8 as an Effective Adjuvant Therapy for Hemorrhagic Shock

项目简介：本次SBIR二期项目拟进一步开发重组人 乳脂球表皮生长因子8(rhMFG-E8)作为一种新的有效的辅助治疗 失血性休克，每年导致60,000名美国人死亡。失血性休克导致细胞死亡， 垂死的细胞会释放与损伤相关的分子模式(DAMP)。潮湿会引发炎症， 复合器官损伤导致多器官衰竭，这是发病率和 出血后死亡率。MFG-E8是一种分泌型糖蛋白，能促进死亡细胞的清除， 从而消除了湿气的释放。在我们的初步研究中，我们已经表明，循环中的 MFG-E8在失血小鼠中减少，而His标记的rhMFG-E8辅助治疗 减少炎症，提高失血性休克存活率。因为他标记的生物制品不是 适合在人类患者中使用，我们使用了人类细胞表达系统来产生纯的无标签 重组人MFG-E8，具有良好的生物活性。在失血性休克大鼠模型中，辅助性治疗 无标签的重组人MFG-E8显著减少循环中肝、肺、循环中的凋亡细胞数量 促炎细胞因子水平与活化的中性粒细胞对肺的侵袭。无标签的rhMFG-E8 还可减轻肾脏和肝脏损伤，并将失血性休克存活率从50%提高到80%。 此外，我们还测定了重组人MFG-E8‘S的分布和消除半衰期及其非致癌性。 因此，我们假设无标签的重组人MFG-E8可以进一步发展成为一种新的有效的 失血性休克的辅助治疗。为了推进药物开发，我们将确定无标签 重组人MFG-E8‘S对小鼠脏器损伤及血流动力学参数的影响 提高大鼠出血后存活率的治疗窗口。我们还将研究无标签的rhMFG- E8‘S对大鼠的安全性，并验证其对器官损伤、血流动力学参数和 在失血性休克猪模型中存活。这些研究将提供临界剂量、时间 管理、有效性和安全性信息进一步开发无标签的重组人MFG-E8作为辅助疗法 治疗失血性休克。我们未来的步骤将包括扩大MFG-E8的生产并完成 有效性、ADME和毒代动力学临床前研究。然后我们将提交一份调查性新药(IND) 向FDA申请启动临床试验。我们的最终目标是获得TAG的商业用途- 游离重组人MFG-E8作为失血性休克患者安全有效的复苏佐剂。

PROJECT DESCRIPTION: This SBIR Phase II project proposes to further develop recombinant human milk fat globule epidermal growth factor-factor 8 (rhMFG-E8) as a novel and effective adjuvant therapy for hemorrhagic shock, which kills 60,000 Americans every year. Hemorrhagic shock results in cell death, and dying cells release damage-associated molecular patterns (DAMPs). DAMPs promote inflammation, compounding organ injury to cause multiorgan failure, which is an important cause of morbidity and mortality after hemorrhage. MFG-E8 is a secreted glycoprotein that promotes the clearance of dying cells, thus abrogating the release of DAMPs. In our preliminary studies, we have shown that circulating levels of MFG-E8 are reduced in hemorrhaged mice, and that their adjuvant treatment with His-tagged rhMFG-E8 decreases inflammation and improves hemorrhagic shock survival. Since His-tagged biologics are not suitable for the use in human patients, we used a human cell expression system to produce pure tag-free rhMFG-E8, with superior biological activity. In a rat model of hemorrhagic shock, adjuvant treatment with tag-free rhMFG-E8 significantly reduced the number of apoptotic cells in the liver and lungs, circulating levels of proinflammatory cytokines and pulmonary infiltration by activated neutrophils. Tag-free rhMFG-E8 also attenuated renal and hepatic injury and improved hemorrhagic shock survival from 50% to 80%. Additionally, we determined rhMFG-E8’s distribution and elimination half-lifes and its non-carcinogenicity. Therefore, we hypothesize that tag-free rhMFG-E8 can be further developed as a new and effective adjuvant therapy for hemorrhagic shock. To advance the drug development, we will determine tag-free rhMFG-E8’s dose-dependent beneficial effects on organ injury and hemodynamic parameters, as well as its therapeutic window to improve survival after hemorrhage in the rat. We will also examine tag-free rhMFG- E8’s safety profile in the rat and verify its beneficial effects on organ injury, hemodynamic parameters, and survival in a pig model of hemorrhagic shock. These studies will provide critical dosing, time of administration, efficacy, and safety information to further develop tag-free rhMFG-E8 as an adjuvant therapy for hemorrhagic shock. Our future steps will include scaling up production of MFG-E8 and completing efficacy, ADME, and toxicokinetic preclinical studies. We will then file an investigative new drug (IND) application with the FDA to initiate clinical trials. Our ultimate goal is to obtain commercial utilization of tag- free rhMFG-E8 as a safe and effective resuscitation adjuvant for patients with hemorrhagic shock.