Optimized lactoferrin for treatment of intracerebral hemorrhage

 描述（由申请人提供）：脑出血（ICH）是一个主要的公共卫生问题，在所有中风亚型中死亡率最高。 ICH 是长期残疾的主要原因之一。由于目前尚无 FDA 批准的 ICH 治疗方法，因此寻找治疗这种破坏性医疗状况的新方法至关重要。 乳铁蛋白 (LTF) 是一种众所周知的内源性糖蛋白，具有抗微生物和免疫调节功能，部分是通过其有效隔离游离铁实现的。在我们正在进行的研究中，使用不同的体外细胞培养系统（模拟 ICH 发病机制）和临床相关的 ICH 啮齿动物模型，我们发现乳铁蛋白 (LTF) 具有独特的多效性活性。这使得这种内源性蛋白质作为治疗脑出血的候选药物极具吸引力。由于 LTF 会迅速从循环中去除（T1/2 ~45 分钟），我们开发了一种基于重组人乳铁蛋白 (rhLTF) 和新生儿 Fc 受体 IgG Fc 结构域 (PRC14) 的新型融合蛋白。这种新型分子确实比天然 LTF 具有显着延长的半衰期（5.8 倍）。我们通过确定 PRC14 的最佳治疗剂量并在 ICH 小鼠模型中显示出非凡的治疗窗（24 小时），成功实现了 I 期目标。该 II 期的主要目标是：(i) 根据 GMP 标准验证 PRC14 作为在稳定 CHO (DG44) 细胞中表达的潜在治疗剂的安全性（目标 1），以及 (ii) 使用模拟临床危险因素和 ICH 后不良结果的已知决定因素的动物模型，进一步表征 PRC14 在治疗 ICH 中的治疗效用（目标 2）。 PRC14的进一步商业化将需要一个稳健、可扩展的程序，其中最终材料保留活性并满足动物临床前安全性和毒理学的合规要求。我们将通过根据 GMP 协议在稳定的 CHO 细胞中生产 PRC14 来实现这一目标，并根据 GLP 协议进行以下测试：(i) 药代动力学； (ii) 单剂量安全性研究（毒性）； (iii) 在大鼠中进行重复给药、为期 7 天的安全性研究（毒性）。 PRC14 取自 放大生产（25克）将用于评估早期水肿和长期神经功能缺损：（i）老年雌性和雄性小鼠，（ii）自发性高血压大鼠（SHR），以及（iii）猪 模型允许更大的血肿尺寸。该设计应该为未来的临床研究提供坚实的基础。

 DESCRIPTION (provided by applicant): Intracerebral hemorrhage (ICH) is a major public health problem with the highest mortality rate of all stroke subtypes. ICH is one of the leading causes of long-term disability. Since there are no available FDA- approved therapies for ICH, it is of vast importance to search for new approaches in treatment for this devastating medical condition. Lactoferrin (LTF) is a well-known endogenous glycoprotein with anti-microbial and immunoregulatory functions, in part through its effective sequestration of free iron. In our ongoing research, using different in vitro cell-culture systems (to model ICH pathogenesis) and clinically relevant rodent models of ICH, we discovered that lactoferrin (LTF) has unique pleiotropic activities. This makes this endogenous protein highly attractive as a therapeutic candidate for the treatment of ICH. Because LTF is rapidly removed from the circulation (T1/2 ~45 min), we developed a novel fusion protein based on recombinant human lactoferrin (rhLTF) and Fc domain of IgG for neonatal Fc receptor (PRC14). This novel molecule has indeed a significantly extended half- life (5.8 fold) over native LTF. We successfully accomplished the objectives of Phase I by defining the optimal therapeutic dose of PRC14 and showing extraordinary therapeutic window (24h) in a mouse model of ICH. The primary objective of this Phase II is: (i) to validate the safety of PRC14 as a potential therapeutic expressed in stable CHO (DG44) cells according to GMP standards (Aim 1), and (ii) to further characterize the therapeutic utility of PRC14 in treatment for ICH, using animal models that simulate clinical risk factors and known determinants of poor outcome after ICH (Aim 2). Further commercialization of PRC14 will require a robust, scalable procedure where the final material retains activity and meets compliance requirements for pre-clinical safety and toxicology in animals. We will accomplish this goal with PRC14 being produced in stable CHO cells under GMP protocols and tested according to GLP protocols for: (i) pharmacokinetics; (ii) single dose safety studies (toxicity); and (iii) repeat dose, 7-day safety studies (toxicity) in rats. The PRC14 obtained from scale-up production (25g) will be used to assess early edema and long-term neurological deficit: (i) in aged female and male mice, (ii) in spontaneously hypertensive rats (SHR), and (iii) in a pig model allowing for larger hematoma size. This design should provide the strong foundation for future clinical studies.