Novel molecular therapeutics for cystic fibrosis

囊性纤维化的新型分子疗法

基本信息

批准号：
8315005
负责人：
David E Sterner
金额：
$ 28.28万
依托单位：
PROGENRA, INC.
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-15 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8315005
关键词：
Accounting Address American Anti-Inflammatory Agents Antibiotics Area Bacteria Biological Assay Cell membrane Cell model Cell surface Cells Cessation of life Chemicals Chloride Ion Chlorides Chronic Disease Clinical Trials Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Degradation Pathway Development Disease Diversity Library Drug Delivery Systems Drug Industry Effectiveness Endoplasmic Reticulum Enhancers Enzyme Inhibitor Drugs Enzyme Inhibitors Enzyme-Linked Immunosorbent Assay Fingers Gastrointestinal tract structure Goals Growth Hereditary Disease Human In Vitro Individual Infection Laboratories Lead Ligase Lung Membrane Modeling Molecular Mucolytics Mucous Membrane Mucous body substance Mutate Mutation Pathology Pathway interactions Patients Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacologic Substance Phase Phenylalanine Plant Roots Positioning Attribute Proteins Regulation Salts Screening procedure Sepsis Sodium Chloride Sorting - Cell Movement Symptoms Technology Therapeutic Therapeutic Agents Ubiquitin Uncertainty Up-Regulation VX-809 Validation Work base counterscreen cystic fibrosis patients disability drug candidate drug discovery high throughput screening improved in vitro Assay in vivo inhibitor/antagonist interest microorganism mortality multicatalytic endopeptidase complex mutant novel pre-clinical protein degradation salt balance septic small molecule trafficking ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): Approximately 70,000 individuals worldwide suffer from the debilitating disease cystic fibrosis (CF), which typically is fatal owing to sepsis in the abnormally viscous mucus of the lung. Various treatments are employed to address symptoms, but an approved drug that acts on the cause of CF is urgently needed. The cause is the ?F508 mutation of the CF (CFTR), which aberrantly regulates levels of salts inside and outside cells, leading to the disease pathologies. The mutation causes improper folding of the CFTR protein, so that most of it is degraded in the endoplasmic reticulum (ER) in the cell post-synthesis (ERAD) and does not reach the cell membrane. CFTR?F508 that does localize to the membrane is partially functional, and drugs called correctors (e.g., VX-809) are being developed to promote trafficking of mutant CFTR to the cell surface. One of the ERAD ubiquitin E3 ligases that target CFTR is the RING ligase gp78, and knockdown of gp78 leads to dramatically increased levels of CFTR?F508 in cells. Moreover, cells containing catalytically dead gp78 are characterized by reduced ubiquitylation of CFTR?F508. Due to upregulation of CFTR?F508 levels, gp78 inhibition has the potential to increase the amount of CFTR at cell surfaces, especially in combination with correctors such as VX-809. It is thus proposed that gp78 is a novel target for developing small molecule inhibitors for treating CF by increasing levels of functional CFTR, thereby improving the regulation of mucus in the lungs of CF patients. Progenra has developed an assay technology that has identified small molecule inhibitors of various RING E3 ligases; these inhibitors are currently undergoing pre-clinical development. It is proposed here to adapt and validate this assay for screening for inhibitors of human gp78 and conduct a pilot screen for small molecules that can be developed as therapeutic agents for CF. Autoubiquitylation and substrate ubiquitylation assays will be developed, 40,000 compounds from a diversity library will be screened, and suitable inhibitors will be identified based on potency and selectivity as determined by counter-screening of the hits. An appropriate cellular model will be established and cell proof of concept studies initiated on selected inhibitors. In Phase II, more extensive screening will be conducted and selected lead gp78 inhibitors will undergo preclinical development (chemical optimization, DMPK, in vivo efficacy studies) for treatment of CF singly or in combination with correctors such as VX-809. PUBLIC HEALTH RELEVANCE: Although various treatments are employed to address symptoms of the debilitating disease cystic fibrosis (CF), an approved drug that acts on the cause of CF is urgently needed. A protein called CFTR is responsible for maintaining proper salt balance at cell membranes and in CF it is mutated and degraded, leading to imbalance and overly mucous membranes that become septic leading to mortality. Progenra proposes to develop a small molecule inhibitor of the enzyme (gp78) that degrades this mutated protein to restore salt balance and achieve therapeutic benefit.

描述（由申请人提供）：全世界约有70,000个人患有使人衰弱的疾病囊性纤维化（CF），通常由于败血症而致命肺部异常粘性粘液。采用各种治疗方法来解决症状，但迫切需要根据CF原因作用的批准药物。原因是CF（CFTR）的F508突变，该突变异常调节内部和外部细胞外部盐的水平，导致疾病的病理。该突变会导致CFTR蛋白的不当折叠，因此大部分在细胞合成后（ERAD）中的内质网（ER）中降解，并且不到达细胞膜。 cftr？f508定位在膜上是部分功能，并且正在开发称为校正器的药物（例如VX-809），以促进将突变CFTR的运输促进对细胞表面的运输。靶向CFTR的ERAD泛素E3连接酶之一是环连接酶GP78，而GP78的敲低导致细胞中CFTR？F508的水平急剧增加。此外，含有催化死亡GP78的细胞的特征是CFTR？F508的泛素化降低。由于CFTR？F508水平的上调，GP78的抑制可能会增加细胞表面的CFTR量，尤其是与诸如VX-809之类的校正器结合使用。因此，有人提出，GP78是通过增加功能CFTR水平来开发用于治疗CF的小分子抑制剂的新靶标，从而改善了CF患者肺中粘液的调节。后代已经开发了一种测定技术，该技术已经鉴定出各种环E3连接酶的小分子抑制剂。这些抑制剂目前正在进行临床前发展。这里提议适应和验证该测定法以筛选人类GP78的抑制剂，并为小分子进行试验筛选，该分子可以作为CF的治疗剂开发。将开发自动素化和底物泛素化测定法，将筛选40,000种来自多样性库中的化合物，并根据命中率反应确定的效力和选择性来鉴定合适的抑制剂。将建立适当的细胞模型，并在选定的抑制剂上启动概念研究的细胞证明。在II阶段，将进行更广泛的筛查，选定的铅GP78抑制剂将经历临床前开发（化学优化，DMPK，体内疗效研究），以单独处理CF或与VX-809等校正器进行治疗。公共卫生相关性：尽管采用了各种治疗方法来解决使人衰弱的疾病囊性纤维化（CF）的症状，这是一种批准的药物，其作用于CF原因。一种称为CFTR的蛋白质负责在细胞膜处维持适当的盐平衡，在CF中，它被突变和降解，导致失衡和过度粘膜膜，导致死亡率。后代提议开发酶（GP78）的小分子抑制剂，该酶（GP78）降解这种突变的蛋白质以恢复盐平衡并获得治疗益处。