Development of human thioredoxin as a mucolytic therapy for Cystic Fibrosis

开发人硫氧还蛋白作为囊性纤维化的粘液溶解疗法

• 批准号: 8310904
• 负责人: Peter B Heifetz
• 金额: $ 18.31万
• 依托单位: ORPRO THERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2014-03-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8310904
• 关键词: Active Sites; Adrenal Cortex Hormones; Aerosols; Affect; Affinity; Agonist; Amino Acids; Antibiotics; Biological Assay; Breathing; Buffers; Caucasians; Caucasoid Race; Cloning; Codon Nucleotides; Complex; Cystic Fibrosis; DNA Sequence; Deoxyribonuclease I; Deoxyribonucleases; Development; Devices; Disease; Disulfides; Drug Formulations; Endotoxins; Enzymes; Escherichia coli; Excipients; Goals; Human; In Vitro; Individual; Infection; Inherited; Laboratories; Laboratory Study; Leukocyte Elastase; Lung Inflammation; Lung diseases; Mammalian Cell; Measures; Medical; Methods; Mucins; Mucolytics; Mucous body substance; NMR Spectroscopy; Nebulizer; Nucleic Acids; Oxidation-Reduction; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Phase; Population; Powder dose form; Production; Proteins; Recombinant Proteins; Recombinants; Relative (related person); Research; Respiratory physiology; Safety; Saline; Solubility; Solutions; Solvents; Spectroscopy, Fourier Transform Infrared; Sputum; Sulfhydryl Compounds; System; Therapeutic; Thioredoxin; Treatment Protocols; United States; Validation; Variant; Wit; analytical method; aqueous; base; chemical property; crosslink; cystic fibrosis patients; drug development; effective therapy; efficacy testing; fluidity; human TXN protein; improved; milligram; mutant; novel; preclinical study; protein expression; recombinant human DNase; standard of care; synthetic construct; therapy design; vector; viscoelasticity

DESCRIPTION (provided by applicant): Effective management of obstructive pulmonary diseases, especially Cystic Fibrosis (CF), represents a significant unmet medical need. Such diseases are characterized by thickened mucus that contributes to impaired lung function. This mucus barrier is composed primarily of cross-linked chains of proteins (mucins) complexed with nucleic acids released as a consequence of persistent lung inflammation and infection. In the case of CF, an autosomal recessive disease affecting approximately 30,000 patients in the US and 70,000 world-wide, current therapies designed to increase the fluidity of mucus and facilitate clearance involve the use of hypertonic saline inhalation to thin secretions and DNase enzymes (dornase alfa, trademarked as Pulmozyme") to digest mucus nucleic acids. In addition, half of patients with CF take four different medications, while about a quarter take upwards of six drugs including short-acting beta agonists, mucolytics, antibiotics, and inhaled and intranasal corticosteroids. There is consequently a great need to reduce treatment burden by developing formulations and delivery regimes that are compatible with existing therapies and delivery devices and that are effective across a broader percentage of CF patients. OrPro Therapeutics, Inc. is developing thioredoxins, a new class of mucolytic protein drugs with high efficacy and safety which specifically disrupt the thiol linkages between crosslinked mucin proteins. Reduced recombinant human thioredoxin-1 (rhTrx) has been demonstrated to be a highly potent liquefier of human CF sputum in laboratory studies. Trx is a small, stable non-glycosylated redox protein suitable for manufacturing in a range of prokaryotic and eukaryotic expression hosts and does not require mammalian cell expression. Our overall goal is to establish a formulation strategy for rhTrx that will support development of this molecule as a new and more efficacious mucolytic therapy for cystic fibrosis. A key component of such a therapy is a drug formulation that accomplishes the objectives of maintaining the protein in a stable, chemically reduced state (essential for its mucolytic and anti-protease functions) as well as facilitating compatibility wit modern, highly efficient aerosol delivery systems that minimize the treatment burden on CF patients. For this Phase I application, we plan to focus on laboratory-scale production of purified proteins (both native rhTrx control and a single amino-acid mutant variant, r(Cys)hTrx, having the potential to enhance safety and efficacy), physical characterization of these proteins in order to assess their chemical properties and relative activities, and initial preformulation studies to evaluate the suitability of thioredoxin for delivery via a range of nebulization devices. If the Ais of this project are achieved, key validation of the concept of disulfide reduction for mucolytic drug development will be established, enabling a Phase II project with the goal of focused development of specific formulations that will support preclinical studies leading to human trials of potentially safer and more efficacious treatment regimens that utilize improved, patient-friendly delivery devices. PUBLIC HEALTH RELEVANCE: There remains a critical unmet need for safe and effective treatment for Cystic Fibrosis (CF), the most common inherited lethal disease in Caucasian populations affecting more than 30,000 individuals in the United States. Our overall goal is to develop a new and improved mucus-disrupting (mucolytic) therapy for CF that is highly efficacious and which does not impose excessive treatment burdens on CF patients. We will accomplish this through development of optimal formulations of inhaled human thioredoxin, a recombinant protein mucolytic with a novel mechanism of action that has the potential to benefit a broader range of CF patients than current therapies.

描述（由申请人提供）：阻塞性肺病，特别是囊性纤维化（CF）的有效治疗代表了一个重大的未满足的医疗需求。此类疾病的特点是粘液增稠，导致肺功能受损。这种粘液屏障主要由交联的蛋白质链（粘蛋白）与持续肺部炎症和感染释放的核酸复合而成。就 CF 而言，这是一种常染色体隐性遗传病，影响了美国约 30,000 名患者和全世界 70,000 名患者，目前旨在增加粘液流动性和促进清除的治疗方法包括使用高渗盐水吸入来稀释分泌物和 DNase 酶（dornase） alfa，商标为 Pulmozyme”）来消化粘液核酸。此外，一半的 CF 患者服用四种不同的药物，而大约四分之一的患者服用以上药物六种药物，包括短效β受体激动剂、粘液溶解剂、抗生素以及吸入和鼻内药物 皮质类固醇。因此，非常需要通过开发与现有疗法和递送装置兼容并且对更广泛的 CF 患者有效的制剂和递送方案来减轻治疗负担。 OrPro Therapeutics, Inc. 正在开发硫氧还蛋白，这是一类新型粘液溶解蛋白药物，具有高效和安全性，可特异性破坏交联粘蛋白之间的硫醇连接。实验室研究表明，还原型重组人硫氧还蛋白-1 (rhTrx) 是一种高效的人 CF 痰液液化剂。 Trx 是一种小型、稳定的非糖基化氧化还原蛋白，适合在一系列原核和真核表达宿主中生产，不需要哺乳动物细胞表达。我们的总体目标是建立 rhTrx 的配方策略，以支持该分子作为一种新的、更有效的囊性纤维化粘液溶解疗法的开发。这种疗法的一个关键组成部分是药物制剂，该制剂能够实现将蛋白质维持在稳定的化学还原状态（对其粘液溶解和抗蛋白酶功能至关重要）以及促进与现代高效气雾剂输送系统的相容性的目标最大程度地减轻 CF 患者的治疗负担。对于这一阶段的应用，我们计划重点关注纯化的实验室规模生产 蛋白质（天然 rhTrx 对照和单氨基酸突变变体 r(Cys)hTrx，具有增强安全性和功效的潜力），这些蛋白质的物理表征，以便 评估其化学性质和相关活性，并进行初始预制剂研究，以评估硫氧还蛋白通过一系列雾化装置输送的适用性。如果该项目的 AIS 得以实现，则将建立对粘液溶解药物开发中二硫键还原概念的关键验证，从而启动 II 期项目，其目标是重点开发特定制剂，以支持临床前研究，从而进行潜在的人体试验。更安全、更有效的治疗方案，利用改进的、患者友好的输送设备。 公共卫生相关性：囊性纤维化 (CF) 是白种人群体中最常见的遗传性致命疾病，在美国影响超过 30,000 人，对安全有效治疗的需求仍然未得到满足。我们的总体目标是开发一种新的、改进的粘液破坏（粘液溶解）治疗方法，该方法非常有效，并且不会给 CF 患者带来过多的治疗负担。我们将通过开发吸入人硫氧还蛋白的最佳配方来实现这一目标，这是一种重组蛋白粘液溶解剂，具有新颖的作用机制，与现有疗法相比，有可能使更广泛的 CF 患者受益。