Repurposing an FDA approved Drug, B-Raf Kinase Inhibitor Dabrafenib for Protection from Cisplatin- and Noise- induced Hearing Loss

重新利用 FDA 批准的药物 B-Raf 激酶抑制剂 Dabrafenib 来预防顺铂和噪音引起的听力损失

• 批准号: 10322750
• 负责人: Tal Teitz
• 金额: $ 41.45万
• 依托单位: CREIGHTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10322750
• 关键词: Affect; Aging; Anti-Inflammatory Agents; Antioxidants; Auditory; BRAF gene; Benchmarking; Biological; Biological Assay; Biological Markers; Cancer Patient; Cell Death; Cell Line; Cell physiology; Cells; Cessation of life; Cisplatin; Clinic; Clinical; Clinical Research; Clinical Trials; Cochlea; Data; Dexamethasone; Dose; FDA approved; Goals; Hair Cells; Human; Inner Supporting Cell; Knockout Mice; Labyrinth; Lead; Lethal Dose 50; MAP2K1 gene; MAPK3 gene; Malignant Neoplasms; Malignant neoplasm of lung; Medical; Medical Genetics; Methionine; Modeling; Molecular; Molecular Target; Morphology; Mus; Neuroblastoma; Noise; Noise-Induced Hearing Loss; Non-Small-Cell Lung Carcinoma; Oral; Outer Hair Cells; Pathway interactions; Patients; Performance; Persons; Pharmaceutical Preparations; Phosphorylation; Prevention; Property; Protein Kinase; Proto-Oncogene Proteins B-raf; Protocols documentation; Publishing; Raf Kinase Inhibitor; Regimen; Research; Resistance; Role; Route; Signal Transduction; Stress; Supporting Cell; Testing; Therapeutic; Therapeutic Index; Time; Toxic effect; Translations; Tumor Cell Line; Up-Regulation; Work; Zebrafish; antineoplastic antibiotics; cancer therapy; chemotherapy; cisplatin induced hearing loss; conditional knockout; drug repurposing; ebselen; effectiveness testing; experimental study; hearing impairment; high throughput screening; in vivo; inhibitor; kinase inhibitor; lateral line; lung small cell carcinoma; melanoma; mouse model; neuromast; noise exposure; otoprotectant; ototoxicity; pharmacokinetics and pharmacodynamics; postnatal; pre-clinical; prevent; prevent hearing loss; small molecule; small molecule inhibitor; sodium thiosulfate; standard care; standard of care; therapeutic target; tumor

Abstract Hearing loss caused by noise, aging and chemotherapy affects seven hundred million people worldwide, but there are no FDA-approved drugs to prevent it. This research will test the potential to repurpose a small molecule BRAF inhibitor, dabrafenib (TAFINLAR), an FDA-approved drug for several cancers, for new use in preventing cisplatin-induced hearing loss. Dabrafenib was a top hit in our unbiased high-throughput screens of 4,385 bioactive compounds and 187 specific kinase inhibitors for cisplatin-induced cell-death protection in an inner ear cell line. We found that dabrafenib fully protected the outer hair cells against cisplatin toxicity in mouse cochlear explants with IC50 of 30 nM and an excellent therapeutic index (LD50/IC50) of >2000. Mechanistically, we identified dabrafenib and three additional BRAF inhibitors, two MEK1/2 inhibitors, and an ERK1/2 inhibitor immediately downstream of BRAF in the cellular pathway, supporting the role of BRAF in cisplatin-induced hair cell death. Cisplatin treatment of the inner ear cell line caused upregulation of phospho- BRAF, phospho-MEK1/2 and phospho-ERK1/2 that was inhibited by co-treatment with dabrafenib. Moreover, cisplatin treatment of cochlear explants or noise exposure in vivo caused up-regulation of phospho-ERK1/2 short time after damage in supporting cells (inner phalangeal and Deiters’ cells) that was mitigated by dabrafenib treatment. Furthermore, at 100 nM dabrafenib protected zebrafish lateral line neuromasts from cisplatin-induced death in vivo and, importantly, significant protection was achieved with oral delivery of dabrafenib for three consecutive days in mouse models against cisplatin-induced hearing loss. The daily dose of dabrafenib administered to the mice was in the range approved for long-term human treatment. In this proposal, we will test the protection provided by dabrafenib for cisplatin-induced hearing loss in a multiple low dose cisplatin regimen that mimics closely the cisplatin treatment of cancer patients in the clinic. Functional auditory performance and inner ear morphology will be assessed. Two doses of dabrafenib will be tested to evaluate the therapeutic window of the drug in vivo. We will also determine dabrafenib’s interference with cisplatin tumor killing efficacy in tumor cell lines and mouse tumor models in which cisplatin is the standard treatment, neuroblastoma and lung cancer. We will confirm BRAF kinase is the molecular target of dabrafenib in cisplatin-induced hearing loss by generating a supporting cell specific conditional knockout mouse in which BRAF is specifically deleted in the supporting cells of the inner ear starting at postnatal day 28 and testing its resistance to cisplatin. Supporting cells’ ERK phosphorylation can serve as an in vivo biomarker for cisplatin damage and evaluating treatment with BRAF inhibitors and can be utilized to determine dabrafenib’s PK/PD properties. Our study will reveal a new cellular pathway and molecular target BRAF kinase for otoprotection and will provide the crucial data needed for advancing dabrafenib to clinical trials in humans for prevention of cisplatin-induced hearing loss.

抽象的 噪音，衰老和化学疗法造成的听力损失会影响全球7亿人，但 没有FDA批准的药物可以防止它。这项研究将测试重现小的潜力 分子BRAF抑制剂Dabrafenib（Tafinlar），一种用于多种癌症的FDA批准药物，用于新用途 防止顺铂引起的听力损失。 dabrafenib在我们公正的高通量屏幕中受到了最大的打击 4,385种生物活性化合物和187种特异性激酶抑制剂，用于顺铂诱导的细胞死亡保护 内耳细胞系。我们发现dabrafenib完全保护外毛细胞免受顺铂的毒性 小鼠人工耳蜗外植体，IC50为30 nm，出色的治疗指数（LD50/IC50）为> 2000。 从机械上讲，我们确定了dabrafenib和另外三个BRAF抑制剂，两个MEK1/2抑制剂，一个 ERK1/2抑制剂立即在BRAF的蜂窝途径中下游，支持BRAF在 顺铂诱导的毛细胞死亡。内耳细胞系的顺铂治疗导致磷酸的上调 Braf，Phosho-Mek1/2和磷酸化eRK1/2通过与dabrafenib共治疗抑制。而且， 在体内对耳蜗外植体或噪声暴露的顺铂治疗导致磷酸-ERK1/2的上调 损坏后不久，支撑细胞（内向司司兰格和脱发细胞）受到了缓解 dabrafenib治疗。此外，在100 nm dabrafenib处保护斑马鱼侧线神经瘤 顺铂诱导的体内死亡，重要的是，通过口服递送实现了明显的保护 在小鼠模型中连续三天反对顺铂引起的听力损失。每日剂量 给小鼠施用的dabrafenib的长期治疗范围内。在这个 提案，我们将测试Dabrafenib为顺铂引起的听力损失的保护 剂量的顺铂方案是临床中癌症患者的顺铂治疗。功能 听觉性能和内耳形态将被评估。两剂dabrafenib将被测试到 评估体内药物的治疗窗口。我们还将确定dabrafenib的干扰 顺铂肿瘤在肿瘤细胞系和小鼠肿瘤模型中杀死效率，其中顺铂是标准 治疗，神经母细胞瘤和肺癌。我们将确认BRAF激酶是dabrafenib的分子靶标 在顺铂诱导的听力损失中，通过产生支撑细胞的特定条件敲除小鼠 BRAF在出生后第28天开始在内耳的支撑细胞中专门删除，并测试其 对顺铂的抗性。支持细胞的ERK磷酸化可以作为顺铂的体内生物标志物 用BRAF抑制剂损坏和评估治疗，可以用于确定Dabrafenib的PK/PD 特性。我们的研究将揭示一种新的细胞途径和分子靶标BRAF激酶用于局部保护 并将提供将dabrafenib推进到人类临床试验所需的关键数据，以预防 顺铂引起的听力损失。