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，在所有卒中亚型中死亡率最高。ICH是导致长期残疾的主要原因之一。由于没有FDA批准的ICH治疗方法，因此寻找治疗这种毁灭性疾病的新方法非常重要。 乳铁蛋白（LTF）是一种具有抗微生物和免疫调节功能的内源性糖蛋白，部分通过其有效螯合游离铁。在我们正在进行的研究中，使用不同的体外细胞培养系统（以模拟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ICH的治疗候选物非常有吸引力。由于LTF从循环中迅速清除（T1/2 ~45 min），我们开发了一种基于重组人乳铁蛋白（rhLTF）和新生儿Fc受体（PRC 14）IgG Fc结构域的新型融合蛋白。这种新分子确实具有比天然LTF显著延长的半衰期（5.8倍）.我们通过定义PRC 14的最佳治疗剂量并在ICH小鼠模型中显示出非凡的治疗窗口（24小时），成功地实现了I期的目标。该II期研究的主要目标是：（i）根据GMP标准（目标1）验证PRC 14作为在稳定CHO（DG 44）细胞中表达的潜在治疗药物的安全性，以及（ii）进一步表征PRC 14在治疗ICH中的治疗效用，使用模拟临床风险因素和ICH后不良结果的已知决定因素的动物模型（目标2）。 PRC 14的进一步商业化将需要一个稳健的、可扩展的程序，其中最终材料保留活性并符合动物临床前安全性和毒理学的合规性要求。我们将根据GMP方案在稳定的CHO细胞中生产PRC 14，并根据GLP方案在大鼠中进行以下测试来实现这一目标：（i）药代动力学;（ii）单次给药安全性研究（毒性）;和（iii）重复给药，7天安全性研究（毒性）。PRC 14从 将使用放大生产（25 g）来评估早期水肿和长期神经功能缺损：（i）在老年雌性和雄性小鼠中，（ii）在自发性高血压大鼠（SHR）中，和（iii）在猪中 模型允许更大的血肿尺寸。该设计为未来的临床研究提供了坚实的基础。

 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.