Optimized lactoferrin for treatment of intracerebral hemorrhage

优化乳铁蛋白治疗脑出血

• 批准号: 9016473
• 负责人: Jaroslaw Aronowski
• 金额: $ 4.76万
• 依托单位: PHARMAREVIEW CORPORATION
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9016473
• 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(申请人提供)：脑出血(ICH)是一个主要的公共卫生问题，在所有中风亚型和长期残疾中死亡率最高。由于目前还没有FDA批准的治疗脑出血的方法，因此为这种疾病建立有效的治疗方法是非常重要的。脑出血后，沉积的血液最初通过压缩脑组织(质量效应)受损，然后通过有毒的化学效应 脑组织上的血肿成分。后者包括溶血产物(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、血脑屏障破坏和致死性脑水肿。乳铁蛋白(LTF)是一种广为人知的内源性糖蛋白，具有抗微生物和免疫调节功能，部分是通过有效地隔离游离铁来实现的。在体外和体内脑出血模型中，我们的新发现表明：(1)LTF具有多效性作用机制，可以有效地对抗脑出血发病机制的多因素；(2)在实验性脑出血模型中，它提供了强大的保护作用；(3)一种新的优化LTF-人LTF(HLTF)与新生儿免疫球蛋白Fc受体(PRC14)的融合-比单独使用hLTF更有效。该项目的总体目标是开始使用PRC14治疗脑出血的优化进程。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对PRC14 T1/2的分析来启动PRC14的临床前开发。目的1.利用成年雄性小鼠生产PRC14并确定最佳剂量和治疗时间窗。目标1a。优化并在中国仓鼠卵巢细胞(CHO)中表达PRC14。重点将是扩大PRC14的生产。验收标准：我们将生产和提纯(99%纯度)足够数量的PRC14，用于I期研究(100 Mg)。目标1b。评估PRC14对小鼠的疗效。我们将测试PRC14的剂量范围为0.1 mg/kg-10 mg/kg，治疗窗口为3小时、12小时、24小时和48小时。接受标准：神经功能改善20%，治疗时间窗为3h，与赋形剂相比(p&lt；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19-21盲目的研究人员进行。目的：研究PRC14在小鼠体内的半衰期和生物利用度。这个目标的目标是建立在大脑循环中8小时范围内的PRC14水平(生物利用池)，这相当于自然LTF的10倍半衰期。我们将使用山羊抗人LF抗体，用ELISA法检测血清和脑实质中PRC14的水平。接受标准：PRC14的半衰期将比天然LF延长至少2倍，并将至少与天然LTF一样有效地到达脑组织。

项目成果

Lactoferrin and hematoma detoxification after intracerebral hemorrhage.

• DOI: 10.1139/bcb-2020-0116
• 发表时间: 2020-09
• 期刊: Biochemistry and cell biology = Biochimie et biologie cellulaire
• 作者: Xiurong Zhao;M. Kruzel;J. Aronowski

Beneficial Role of Neutrophils Through Function of Lactoferrin After Intracerebral Hemorrhage.

• DOI: 10.1161/strokeaha.117.020544
• 发表时间: 2018-05
• 期刊: Stroke
• 影响因子: 8.3
• 作者: Zhao X;Ting SM;Sun G;Roy-O'Reilly M;Mobley AS;Bautista Garrido J;Zheng X;Obertas L;Jung JE;Kruzel M;Aronowski J
• 通讯作者: Aronowski J