A Novel Thioredoxin Mimetic Prodrug for Prevention of Bronchopulmonary Dysplasia

一种用于预防支气管肺发育不良的新型硫氧还蛋白模拟前药

• 批准号: 8586510
• 负责人: PRAKASH G JAGTAP
• 金额: $ 37.8万
• 依托单位: RADIKAL THERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-05 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8586510
• 关键词: 3-nitrotyrosine; Acetylcysteine; Acute; Acute Lung Injury; Alveolar; Alveolus; Animal Model; Anti-Inflammatory Agents; Anti-inflammatory; Antioxidants; Applications Grants; Attenuated; Autopsy; Biological Models; Blood; Blood Vessels; Breathing; Bronchopulmonary Dysplasia; Cells; Chronic; Chronic lung disease; Clinical; Collagen; Collection; Colorado; Cultured Cells; Cytoplasmic Protein; Development; Dose; Drug Controls; Drug Kinetics; Drug Targeting; Equilibrium; Euthanasia; Evaluation; Evolution; Exhibits; Fetal Lung; Fibrosis; Functional disorder; Gases; Gene Expression; Gestational Age; Glutathione; Glutathione Disulfide; Growth; Heart; Histologic; Hospitalization; Hyperoxia; Impairment; In Vitro; Infant; Infiltration; Inflammatory; Injury; Interleukin-6; Label; Laboratories; Life; Lipid Peroxidation; Low Birth Weight Infant; Lung; Lung diseases; Macrophage Inflammatory Protein-1; Malondialdehyde; Medical; Modeling; Mus; Neonatal; Neutrophil Infiltration; Newborn Infant; Newborn Respiratory Distress Syndrome; Nuclear Translocation; Ovalbumin; Oxidation-Reduction; Patients; Peptides; Peroxonitrite; Pharmacologic Substance; Phase; Placebo Control; Plasma; Play; Pneumonia; Poly Adenosine Diphosphate Ribose; Powder dose form; Predisposition; Premature Birth; Premature Infant; Prevention; Prodrugs; Production; Proteins; Pulmonary Fibrosis; Pulmonary Valve Insufficiency; Randomized; Rattus; Reduced Glutathione; Reducing Agents; Relative (related person); Resuscitation; Right Ventricular Hypertrophy; Role; Route; Severities; Small Business Innovation Research Grant; Smooth Muscle; Solutions; Stress; Structure; Structure of parenchyma of lung; Sulfhydryl Compounds; Superoxide Dismutase; Superoxides; TNF gene; Testing; Therapeutic; Thioredoxin; Time; Tissues; Trichrome stain; United States National Institutes of Health; Universities; Ventricular; Water; base; clinically relevant; cysteinylproline; cytokine; density; esterase; in vivo; innovation; interstitial; intraperitoneal; liquid formulation; lung injury; mimetics; mimicry; mortality; muscle hypertrophy; neutrophil; novel; oxidant stress; oxidation; premature; premature lungs; prolyl-serine; prospective; prototype; public health relevance; pulmonary arterial hypertension; pup; residence; respiratory; respiratory distress syndrome; seryl-proline; small molecule; thioester

DESCRIPTION (provided by applicant): Radikal Therapeutics (RTX) is developing a novel pharmaceutical therapy for prematurity that targets a developmental deficiency in thioredoxin (Trx), a protein that serves as a central regulator of cellular reductant status. Trx is expressed t levels in the fetal lung too low to adequately counter the redox stress of extrauterine life, and i thus relatively deficient in extreme prematurity. The low level of Trx in alveoli is thought to pla a major role in the susceptibility of very low birthweight (VLBW) premature infants to neonatal respiratory distress syndrome (RDS). Our current understanding of RDS and its evolution into bronchopulmonary dysplasia (BPD) is consistent with a mechanism of injury produced by an excess of superoxide within the lung parenchyma. Compounding this increase in superoxide production is the developmental deficiency of anti-oxidant proteins, such as superoxide dismutase and Trx, that are present in insufficient quantity at an early gestational age. Our innovation is based upon the administration of a novel agent, R-908, a prodrug of a Trx mimetic (R-901) that substitutes for the deficient Trx protein and restores intracellular redox status. R-901 is a thiol-rich tripeptide closely analogous to the native conserved Trx motif and exhibits extraordinary potency: R- 901 is 450-fold more potent than N-acetyl cysteine in the protection of cultured cells from oxidant stress. In a murine ovalbumin model of pulmonary inflammation model, R-901 reduced histologic injury, diminished neutrophil infiltration, attenuated tissue oxidation, blocked pro-inflammatory cytokine expression and nuclear translocation of NF-?B, diminished the degradation of the anti-inflammatory cytoplasmic protein I?B¿, and restored the balance of reduced and oxidized forms of glutathione. In a murine LD60 model of redox stress, induced by acute Cl2 inhalation, post-insult administration of R-901 eliminated all mortality. To overcome shelf instability of the free thiol groups of R-901, we have invented a stable dithioester prodrug (R-908) that releases R-901 in vivo. R-908 will now undergo evaluation in a clinically-relevant rat pup model of BPD. Aim #1: Define the pharmacokinetics (PK) of R-908 in newborn rats. We will define the plasma and tissue PK profile of R-908, and its metabolite R-901, in 2-day old rat pups. Aim #2: Establish that R-908 attenuates changes of pulmonary vascular and alveolar structure in a hyperoxic model of BPD in neonatal rats; 2-day old rat pups will be subjected to hyperoxia for 10 days. R-908 will be administered over a broad dose range for 3 weeks, a period characterized in this model system by progressive lung fibrosis, pulmonary arterial hypertension (PAH), and hypoalveolarization. Lung tissue taken at necropsy will be analyzed for pulmonary vascular structure and growth, alveolarization, lipid peroxidation (malondialdehyde), reduced and oxidized glutathione (GSH, GSSG), peroxynitrite formation (3-nitrotyrosine), poly(ADP-ribose) formation, fibrosis (Mason Trichrome staining for collagen), R-908 and R-901 levels, and pro-inflammatory gene expression. The heart will undergo morphometric analysis for evidence of PAH (as shown by right ventricular (RV) hypertrophy).

描述（由适用提供）：Radikal Therapeutics（RTX）正在开发一种新型的药物治疗，以靶向硫氧还蛋白（TRX）的发育缺乏，这是一种蛋白质，该蛋白是细胞还原状态的中心调节剂。 TRX在胎儿肺中表达t水平过低，无法充分抵消介绍性生活的氧化还原应力，因此我在极端早产中相对不足。肺泡中TRX的低水平被认为在非常低的出生体重（VLBW）早产儿对新生儿呼吸窘迫综合征（RDS）的敏感性中起主要作用。我们目前对RD的理解及其对支气管肺发育不良（BPD）的进化与肺实质中过量的超氧化物产生的损伤机制一致。抗氧化蛋白的发育缺乏，例如超氧化物歧化酶和TRX，在早期妊娠时代的数量不足。我们的创新基于新型药物R-908的给药，R-908是TRX Mimetic（R-901）的前药，该前药代替了缺陷TRX蛋白并恢复细胞内氧化还原状态。 R-901是一种与天然保守的TRX基序相似的富含硫醇的三肽，并且表现出非凡的效力：R-901在保护培养细胞免受氧化物胁迫的保护中的潜力比N-乙酰基半胱氨酸高450倍。在肺部炎症模型的鼠卵巢蛋白模型中，R-901减少了组织学损伤，中性粒细胞浸润减少，减毒组织氧化，阻塞促炎细胞因子的表达和NF-的核易位，NF-？B？谷胱甘肽。在由急性CL2吸入引起的鼠LD60氧化还原胁迫模型中，R-901的灭火后给药消除了所有死亡率。为了克服R-901的自由硫醇组的架子不稳定，我们发明了稳定 Prodrug（R-908）在体内释放R-901。现在，R-908将在BPD的临床上与大鼠幼犬模型中进行评估。 AIM＃1：定义新生大鼠R-908的药代动力学（PK）。我们将在2天老的大鼠幼犬中定义R-908的血浆和组织PK谱及其代谢物R-901。 AIM＃2：确定R-908减弱了新生大鼠BPD高氧模型中肺血管和肺泡结构的变化； 2天老鼠的幼崽将经历10天的高氧。 R-908将在宽剂量范围内施用3周，在该模型系统中以进行性肺纤维化，肺动脉高压（PAH）和低肺泡化为特征。将分析在尸检时进行的肺组织，分析肺血管结构和生长，肺泡化，脂质过氧化（丙二醛），还原和氧化的谷胱甘肽（GSH，GSSG）（GSH，GSSG），过氧亚硝酸盐形成（3-核霉菌），adp-rostrosine fictrice ribrosore）形成式形式，形成式形式，形成式形式，形成式形式，形成式形式，形成式形式，形成式形式，形成式形式，构成，胶原蛋白），R-908和R-901水平以及促炎基因表达。心脏将进行形态计量分析以证明PAH的证据（如右心（RV）肥大所示）。