Preclinical development of CFTR stabilizers targeting the CAL PDZ domain

针对 CAL PDZ 结构域的 CFTR 稳定剂的临床前开发

• 批准号: 9271954
• 负责人: DEAN R MADDEN
• 金额: $ 49.95万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9271954
• 关键词: Address; Affect; Affinity; Alleles; Alpha Cell; Antibiotics; Apical; Area; Binding; Bioavailable; Biochemical; Biological; Biological Assay; Biological Availability; Biopsy; Breathing; Cell Membrane Permeability; Cells; Chemicals; Chemistry; Chloride Channels; Chronic; Clinical; Clinical Trials; Collaborations; Combined Modality Therapy; Computer Simulation; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Data; Defect; Degradation Pathway; Development; Disease; Disease model; Drug Design; Drug resistance; Engineering; Epithelial; Epithelial Cells; Equilibrium; FDA approved; Failure; Generations; Half-Life; Hereditary Disease; Human; Impairment; In Vitro; Infection; Inflammation; Intestines; Ion Channel; Ion Transport; Ions; Lead; Lethal Dose 50; Life Expectancy; Ligand Binding; Ligands; Liquid substance; Longevity; Lung; Measurement; Mediator of activation protein; Microbial Biofilms; Modeling; Modification; Molecular; Molecular Chaperones; Molecular Target; Morbidity - disease rate; Mucociliary Clearance; Mutate; Mutation; Patients; Peptides; Permeability; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacology; Phase II Clinical Trials; Physiological; Positioning Attribute; Predictive Value; Production; Proteins; Pulmonary Function Test/Forced Expiratory Volume 1; Respiratory physiology; Side; Structure; Temperature; Testing; Therapeutic; Tissues; VX-809; Validation; Work; X-Ray Crystallography; apical membrane; bacterial resistance; base; cystic fibrosis patients; cytotoxicity; cytotoxicity test; design; improved; inhibitor/antagonist; insight; loss of function; mortality; new therapeutic target; novel; peptidomimetics; pre-clinical; preclinical development; programs; public health relevance; rectal; response; restoration; scaffold; small molecule; small molecule inhibitor; structural biology; success; symptom treatment; therapeutic target; translational approach; uptake

DESCRIPTION (provided by applicant): The cystic fibrosis transmembrane conductance regulator (CFTR) is an ion channel that is mutated in patients with cystic fibrosis (CF), disruptin fluid and ion balance in multiple epithelial tissues. In the lung, failure of mucociliary clearance facilitates the establishment of drug-resistant bacterial biofilms, despite advanced antibiotic and pulmonary clearance strategies. As a result, chronic lung infection and inflammation are currently the major causes of CF morbidity and mortality, limiting lifespan to <40 years. ~90% of CF patients carry one or two copies of the �F508 allele, which encodes a protein that is inefficiently folded, shows limited channel activity, and is rapidly degraded. Compounds have been identified that address the folding and channel defects. Neither provides significant benefit as a monotherapy, but in combination they produce significant improvement in lung function (�FEV1 >10%) in 25% of �F508 homozygous patients. To reach more patients and increase the functional response, we propose the early-stage pharmacological validation of a novel translational strategy to address the remaining defect - the degradation of rescued �F508-CFTR. No clinical trials include compounds specifically designed to increase CFTR stability at the apical membrane. Having identified the CFTR-Associated Ligand (CAL) as a key mediator of CFTR degradation, we have localized a critical binding interface, designed peptides that block it, and shown that they act as first-in-class 'stabilizers' of functional �F508-CFTR in polarized CF bronchial epithelial cells. Preclinical advancement of our inhibitor-of- CAL (iCAL) approach is currently limited by lead affinity, delivery, and limited data on the extent of additioal rescue compared to combination therapies currently in clinical trials. Here, we propose to leverage preliminary advances in all three areas. Our new data confirm substantial additivity for a cell-permeable iCAL in concert with VX-809. With a validated target, multiple lead chemistries, a strong suite of functional assays, and structural and biochemical expertise, we propose an integrated structure-activity approach to optimize CAL inhibitors and confirm their therapeutic potential. In Aim 1, using existing peptide inhibitors with state-of-the-art ex vivo CF patient intestinal current measurement and in vitro mucociliary transport assays, we will establish a functional pipeline to evaluate combinations of CAL inhibitors with potentiator and corrector molecules approved or in late-stage clinical trials. In Aim 2, we propose to validate and optimize the bioactivity of iCAL peptides for systemic or inhaled therapies. In Aim 3, building on a suite o biochemical assays, we describe structure-based strategies to improve the affinity and selectivity of small-molecule inhibitors. Together these studies will determine threshold parameters governing CFTR rescue. Having joined forces to produce proof of concept for CFTR stabilizers, our interdisciplinary and tightly coordinated collaboration is well positioned to obtan second-generation CAL inhibitors with demonstrated efficacy and biological tolerability, while developing a compelling portfolio for the further pharmacological development of this novel therapeutic target.

描述（由申请人提供）：囊性纤维化跨膜电导调节器（CFTR）是一种离子通道，在囊性纤维化（CF）患者中发生突变，破坏肺部多个上皮组织中的液体和离子平衡，导致粘液纤毛清除失败。 尽管有先进的抗生素和药物，但仍促进耐药细菌生物膜的建立 因此，慢性肺部感染和炎症是目前 CF 发病和死亡的主要原因，约 90% 的 CF 患者携带一两个拷贝的 F508 等位基因。折叠效率低、通道活性有限且快速降解的蛋白质已被鉴定为解决折叠和通道缺陷的化合物，两者作为单一疗法都没有提供显着的益处，但组合起来可以产生显着的改善。 25% 的 F508 纯合子患者的肺功能（FEV1 >10%）为了覆盖更多患者并提高功能反应，我们建议对一种新的转化策略进行早期药理学验证，以解决剩余的缺陷 - 肺功能的退化。拯救了 �F508-CFTR。没有临床试验包括专门设计用于提高顶膜 CFTR 稳定性的化合物。已确定 CFTR 相关配体 (CAL) 是关键。作为 CFTR 降解的介体，我们定位了一个关键的结合界面，设计了阻断它的肽，并证明它们可以作为极化 CF 支气管上皮细胞中功能性“F508-CFTR”的一流“稳定剂”。与目前临床试验中的联合疗法相比，CAL 抑制剂 (iCAL) 方法目前受到先导亲和力、递送以及额外救援程度数据有限的限制。在这里，我们建议利用所有三个领域的初步进展，证实细胞渗透性 iCAL 与 VX-809 具有显着的相加性，具有经过验证的靶标、多种先导化学物质、一套强大的功能测定和结构。结合生化专业知识，我们提出了一种综合的结构-活性方法来优化 CAL 抑制剂并确认其治疗潜力，在目标 1 中，使用现有的肽抑制剂和最先进的离体 CF 患者肠道电流测量和。在体外粘膜纤毛转运测定中，我们将建立一个功能管道来评估 CAL 抑制剂与已批准或处于后期临床试验的增强剂和校正分子的组合。在目标 2 中，我们建议验证和优化 iCAL 肽的全身或生物活性。在目标 3 中，我们以一系列生化测定为基础，描述了基于结构的策略，以提高小分子抑制剂的亲和力和选择性。这些研究将共同​​确定控制阈值参数。 CFTR 救援。在联手为 CFTR 稳定剂提供概念验证后，我们的跨学科和紧密协调的合作已做好充分准备，以获得具有已证实的功效和生物耐受性的第二代 CAL 抑制剂，同时为该药物的进一步药理学开发开发引人注目的产品组合。新的治疗靶点。

项目成果

How to evaluate the cellular uptake of CPPs with fluorescence techniques: Dissecting methodological pitfalls associated to tryptophan-rich peptides.

如何使用荧光技术评估 CPP 的细胞摄取：剖析与富含色氨酸的肽相关的方法学陷阱。

• 发表时间: 2019
• 作者: Seisel, Quentin;Pelletier, François;Deshayes, Sébastien;Boisguerin, Prisca
• 通讯作者: Boisguerin, Prisca

Highway to Cell: Selection of the Best Cell-Penetrating Peptide to Internalize the CFTR-Stabilizing iCAL36 Peptide.

通往细胞的高速公路：选择最佳细胞穿透肽来内化 CFTR 稳定 iCAL36 肽。

• 发表时间: 2022-04-07
• 影响因子: 5.4
• 作者: Seisel, Quentin;Lakumpa, Israpong;Josse, Emilie;Vivès, Eric;Varilh, Jessica;Taulan;Boisguérin, Prisca
• 通讯作者: Boisguérin, Prisca