Optimized lactoferrin for treatment of intracerebral hemorrhage

优化乳铁蛋白治疗脑出血

基本信息

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
项目类别：
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

项目摘要

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）：脑内出血（ICH）是一个主要的公共卫生问题，所有中风子类型和长期残疾的死亡率最高。由于没有可用的FDA批准疗法用于ICH，因此为这种疾病建立有效的治疗非常重要。在ICH之后，沉积的血液最初是通过压缩脑组织（质量效应）而损害的，然后是通过有害的化学作用脑组织上的血肿成分。后一个过程涉及溶血产物（例如铁），氧化应激，促炎反应，蛋白水解酶介导的细胞外基质修饰，血液脑屏障破坏和致命的脑性水肿的毒性。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，部分通过其有效的自由铁的隔离。 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） - 单独的HLTF更有效。该项目的总体目标是开始使用PRC14作为ICH治疗的优化过程。我们的假设是，这里提出的研究将通过开始剂量优化过程和PRC14 T1/2的分析来启动PRC14的临床前发展。目标1。生产PRC14并使用成年雄性小鼠在ICH中确定最佳剂量和治疗时间窗口。目标1a。在中国仓鼠卵巢细胞（CHO）中优化和生产PRC14。重点是扩大PRC14的生产。接受的标准：我们将在此I阶段研究（100mg）中产生和纯化（99％纯度）足够数量的PRC14。目标1B。评估PRC14在小鼠中的功效。我们将使用3H，12H，24H和48H的治疗窗口在0.1mg-10mg/kg之间测试PRC14的剂量范围。接受的标准：与车辆相比，治疗时间窗口的神经功能的提高20％（p <0.05）。所有研究将遵循NIH指南，严格的随机方法，所有分析将由对治疗分配的研究者进行19-21的盲目进行。目的2。评估小鼠中PRC14的T1/2和生物利用度。该目标的目的是在循环中建立PRC14的水平（可生物利用池）在8小时以上的大脑范围内，相当于10 x半衰期的天然LTF时间。我们将使用山羊抗人LF抗体测量ELISA的血清和脑实质中的PRC14水平。接受的标准：PRC14至少在天然LF上具有2倍的延长半年，并且至少与天然LTF一样有效地到达脑组织。