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（由申请人提供）：脑出血（ICH）是所有中风亚型中死亡率最高且长期致残的重大公共卫生问题。由于目前尚无 FDA 批准的 ICH 治疗方法，因此建立针对这种疾病的有效治疗方法至关重要。 ICH 后，沉积的血液首先通过脑组织受压（质量效应）造成损害，然后通过有毒化学作用造成损害。 脑组织上的血肿成分。后一个过程涉及溶血产物（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。使用 ICH 的体外和体内模型，我们的新发现证明：(1) LTF 具有多效性作用机制，可以有效对抗 ICH 发病机制的多因素，(2) 它在 ICH 实验模型中提供强大的保护作用，(3) 一种新型优化的 LTF - 人 LTF (hLTF) 与新生儿 IgG Fc 受体的融合 (PRC14) - 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 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 PRC14 t1/2 分析来启动 PRC14 的临床前开发。目标 1. 使用成年雄性小鼠生产 PRC14 并确定 ICH 的最佳剂量和治疗时间窗。目标 1a。在中国仓鼠卵巢细胞 (CHO) 中优化并生产 PRC14。重点将是扩大 PRC14 的生产规模。验收标准：我们将生产并纯化（纯度为 99%）足够数量的 PRC14，用于本 I 期研究（100 毫克）。目标 1b。评估 PRC14 在小鼠中的功效。我们将测试 PRC14 的剂量范围为 0.1mg-10mg/kg，治疗窗为 3 小时、12 小时、24 小时和 48 小时。接受标准：与载体相比，治疗时间窗为 3 小时，神经功能改善 20%（p<0.05）。所有研究都将遵循 NIH 指南、RIGOR 随机化方法，并且所有分析都将由对治疗分配不知情的研究人员进行19-21。目标 2. 评估 PRC14 在小鼠体内的 t1/2 和生物利用度。该目标的目标是确定 8 小时范围内大脑循环中 PRC14 的水平（生物可利用库），相当于天然 LTF 半衰期的 10 倍。我们将使用山羊抗人 LF 抗体通过 ELISA 测量血清和脑实质中 PRC14 的水平。接受标准：PRC14 的半衰期至少比天然 LF 长 2 倍，并且至少与天然 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