Optimized lactoferrin for treatment of intracerebral hemorrhage

优化乳铁蛋白治疗脑出血

• 批准号: 8831091
• 负责人: Jaroslaw Aronowski
• 金额: $ 18.91万
• 依托单位: PHARMAREVIEW CORPORATION
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-30 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8831091
• 关键词: Address; Adult; Affect; Animal Model; Anti-Inflammatory Agents; Anti-inflammatory; Antibodies; Antimicrobial Effect; Bioavailable; Biological; Biological Availability; Blinded; Blood; Blood - brain barrier anatomy; Blood Circulation; Blood coagulation; Brain; Brain Edema; Cell Culture System; Cerebral Edema; Cerebral hemisphere hemorrhage; Chemicals; Chinese Hamster Ovary Cell; Clinical; Coagulation Process; Data; Deposition; Development; Disease; Dose; Drug Kinetics; Early Intervention; Enzyme-Linked Immunosorbent Assay; Event; Excision; Experimental Models; Extracellular Matrix; FDA approved; Foundations; Future; Glycoproteins; Goals; Goat; Guidelines; Half-Life; Hematoma; Heme Iron; Hemoglobin; Hemolysis; Hour; Human; Immunoglobulin G; In Vitro; Inflammatory; Inflammatory Response; Iron; Lactoferrin; Lipid Peroxidation; Measures; Mediating; Medical; Microglia; Modeling; Modification; Mus; Nervous System Physiology; Neurologic; Neurons; Oxidative Stress; Pathogenesis; Pathogenicity; Patients; Peptide Hydrolases; Phase; Phenotype; Play; Procedures; Process; Production; Proteins; Public Health; Randomized; Recovery; Research; Research Personnel; Rodent Model; Role; Safety; Secondary to; Series; Serum; Stroke; Testing; Therapeutic; Time; Toxic effect; Treatment Efficacy; United States National Institutes of Health; Validation; antimicrobial; base; brain circulation; brain tissue; clinically relevant; combat; cytotoxic; design; disability; effective therapy; improved; in vivo Model; innovation; killings; macrophage; male; mortality; mouse model; neonatal Fc receptor; neurological recovery; novel; phase 1 study; pre-clinical; process optimization; protective effect; prototype; public health relevance; relating to nervous system; repaired; scale up; treatment strategy

DESCRIPTION (provided by applicant): Intracerebral hemorrhage (ICH) is a major public health problem with highest mortality rate of all stroke subtypes and long-term disability. Since there are no available FDA-approved therapies for ICH it is of enormous importance to establish effective treatment for this medical condition. Following ICH the deposited blood is damaging initially via compression of the brain tissue (mass effect) and then via noxious chemical effect of hematoma components on brain tissue. The latter process involves toxicity of hemolytic products (e.g. iron), oxidative stress, pro-inflammatory responses, proteolytic enzymes-mediated extracellular matrix modification, blood brain barrier disruption and deadly cerebral edema. Lactoferrin (LTF) is a well-known endogenous glycoprotein with anti-microbial and immunoregulatory functions, in part through its effective sequestration of free iron. Using in vitr and in vivo models of ICH our novel findings demonstrate: (1) that LTF possess pleotropic mechanism of action that could effectively combat multifactorial aspects of ICH pathogenesis, (2) that it provides robust protective effect in experimental models of ICH, and that (3) a novel optimized LTF - fusion of human LTF (hLTF) with neonatal Fc receptor for IgG (PRC14) - is more effective than hLTF alone. The overall goal of this project is to begin the optimization process for using PRC14 as treatment for ICH. Our hypothesis is that the studies proposed here will initiate the preclinical development of PRC14 by starting the dose optimization process and the analysis of PRC14 t1/2. Aim 1. To produce PRC14 and to determine the optimal dosing and therapeutic time window in ICH using adult male mice. Aim 1a. To optimize and produce PRC14 in Chinese Hamster Ovary cells (CHO). Focus will be to scale up production of PRC14. Criteria for acceptance: We will produce and purify (99% purity) sufficient quantity of PRC14 for this Phase I study (100mg). Aim 1b. Assess the efficacy of PRC14 in mice. We will test a dose range of PRC14 between 0.1mg-10mg/kg with therapeutic window of 3h, 12h, 24h and 48h. Criteria for acceptance: Improvement of neurological function by 20% with the therapeutic time window of 3h compared to the vehicle (p<0.05). All studies will follow NIH guidelines, RIGOR randomization approach and all analyses will be performed by the investigators blinded regarding the treatment assignments19-21. Aim 2. To assess t1/2 and bioavailability of PRC14 in mice. The goal of this aim is to establish levels (bioavailable pool) of PRC14 in circulation an brain over 8 hours range, which is an equivalent of 10 x half-life time of natural LTF. We will measure PRC14 levels in serum and in the brain parenchyma by ELISA using goat anti-human LF antibodies. Criteria for acceptance: PRC14 will have at least 2-times extended half-life over the native LF and will reach brain tissue at least as efficiently as natural LTF.

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，并具有长期残疾。由于没有FDA批准的ICH治疗方法，因此为这种疾病建立有效的治疗方法非常重要。 在ICH之后，沉积的血液最初通过脑组织的压缩（质量效应）并且然后通过脑组织的有害化学作用（例如，脑出血）而损害。 脑组织上的血肿成分后一过程涉及溶血产物（例如铁）的毒性、氧化应激、促炎反应、蛋白水解酶介导的细胞外基质修饰、血脑屏障破坏和致命的脑水肿。 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。使用ICH的体外和体内模型，我们的新发现表明：（1）LTF具有多效性作用机制，其可有效地对抗ICH发病机制的多因素方面，（2）其在ICH的实验模型中提供稳健的保护作用，以及（3）新的优化的LTF -人LTF（hLTF）与新生儿IgG Fc受体（PRC 14）的融合物-比单独的hLTF更有效。 本项目的总体目标是开始使用PRC 14治疗ICH的优化过程。我们的假设是，本文提出的研究将通过启动剂量优化过程和PRC 14 t1/2分析来启动PRC 14的临床前开发。目标1.使用成年雄性小鼠生产PRC 14并确定ICH中的最佳剂量和治疗时间窗。目标1a。在中国人卵巢细胞（CHO）中优化并生产PRC 14。重点将是扩大PRC 14的生产规模。验收标准：我们将生产并纯化（99%纯度）足量的PRC 14用于本I期研究（100 mg）。目标1b。评估PRC 14在小鼠中的功效。我们将测试PRC 14的剂量范围为0.1mg-10 mg/kg，治疗窗为3小时、12小时、24小时和48小时。接受标准：与媒介物相比，神经功能改善20%，治疗时间窗为3小时（p<0.05）。所有研究将遵循NIH指南、RIGOR随机化方法，并且所有分析将由对治疗分配不知情的研究者进行19 -21。目标2.评估PRC 14在小鼠中的t1/2和生物利用度。该目的的目标是建立在8小时范围内循环脑中的PRC 14的水平（生物可利用库），其相当于天然LTF的10倍半衰期。我们将使用山羊抗人LF抗体通过ELISA测量血清和脑实质中的PRC 14水平。验收标准：PRC 14将具有超过天然LF至少2倍的延长的半衰期，并且将至少与天然LTF一样有效地到达脑组织。