rhMFG-E8 as an Effective Adjuvant Therapy for Hemorrhagic Shock

项目描述：该SBIR II期项目建议进一步开发重组人 乳脂肪球表皮生长因子-8（rhMFG-E8）作为一种新的有效的辅助治疗， 每年有6万美国人死于失血性休克出血性休克导致细胞死亡， 死亡细胞释放损伤相关分子模式（DAMP）。DAMP促进炎症， 复合器官损伤导致多器官衰竭，这是发病的重要原因， 出血后死亡率MFG-E8是一种分泌型糖蛋白，可促进死亡细胞的清除， 从而消除DAMP的释放。在我们的初步研究中，我们已经表明， MFG-E8在衰老小鼠中减少，并且用His标记的rhMFG-E8辅助治疗 减少炎症并提高失血性休克存活率。由于His标记的生物制剂不是 为了适用于人类患者，我们使用人类细胞表达系统来产生纯的无标签的 rhMFG-E8具有良好的上级生物活性。在大鼠失血性休克模型中， 无标签的rhMFG-E8显著减少了肝和肺中凋亡细胞的数量， 促炎细胞因子水平和活化中性粒细胞的肺浸润。无标签rhMFG-E8 还减轻了肾和肝损伤，并将失血性休克存活率从50%提高到80%。 此外，我们还测定了rhMFG-E8的分布和消除半衰期以及其非致癌性。 因此，我们假设无标签rhMFG-E8可以进一步开发为一种新的有效的 出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 rhMFG-E8对器官损伤和血流动力学参数的剂量依赖性有益作用，以及其 改善大鼠出血后存活率的治疗窗。我们还将检查无标签rhMFG- E8在大鼠中的安全性特征，并验证其对器官损伤、血流动力学参数和 在猪失血性休克模型中存活。这些研究将提供关键剂量、 进一步开发无标签rhMFG-E8作为辅助治疗的给药、疗效和安全性信息 出血性休克我们未来的步骤将包括扩大MFG-E8的生产规模， 疗效、ADME和毒代动力学临床前研究。然后我们将提交研究性新药（IND） 向FDA申请启动临床试验。我们的最终目标是获得标签的商业利用- 游离rhMFG-E8是一种安全有效的失血性休克复苏辅助剂。

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.