LEAD OPTIMIZATION AND TARGET VALIDATION OF NEXT GENERATION PYRIMIDINE-BASED UTROPHIN UPREGULATORS FOR DUCHENNE MUSCULAR DYSTROPHY

LOOk-UP旨在开发一种最近发现的卡巴肼-嘧啶utrophin上调剂，即OX 01914，以产生一种用于杜氏肌营养不良症（DMD）的最佳口服小分子药物，DMD是一种罕见的遗传性肌肉消耗性致命疾病。为此，罗素教授和我设计了一个多学科和互补的体外方法，依靠药物化学，体外高通量（HT）ADME-T分析，化学蛋白质组学和一流的分子生物学技术。通过这些方法，我将能够进行：1）对OX 01914进行先导优化，以确定一个安全、稳定、有效的先导候选药物用于临床前研究; 2）进行基于化学蛋白质组学的靶向修饰研究，以确认其对线粒体ATP合成酶外周柄亚基B（ATP 5 F1）蛋白的调节作用，该蛋白先前被确定为OX 01914的潜在靶点; 3）研究utrophin调控的分子机制。到目前为止，DMD仍然没有治愈的方法。上调utrophin是一种内源性安全的肌营养不良蛋白，代表了产生DMD疾病修饰和可获得治疗的最有价值的策略之一，适用于所有患者，无论基因突变如何。在这些前提下，该MSCA项目有以下目标：工作包1。使用多组分反应快速生成OX 01914合成类似物的扩展化学文库。工作包2.通过体外活性评估和新生成文库的ADME-T谱分析，鉴定安全且代谢稳定的先导候选药物。工作包3.使用化学蛋白质组学方法确认DMD文库的ATP 5 F1靶点接合。工作包4.通过RNA测序和蛋白质组分析，并利用分子生物学技术研究utrophin的特异性调控途径，为utrophin的调控机制提供新的认识。

LOOk-UP aims at progressing a recently identified carbahydrazide-pyrimidine utrophin upregulator, namely OX01914, to generate a best-in-classorally administrable small molecule drug for the Duchenne Muscular Dystrophy (DMD), a rare genetic muscle-wasting fatal disease. To this end,Prof Russell and I designed a multidisciplinary and complementary in vitro approach leaning on medicinal chemistry, in vitro High-Throughput (HT)ADME-T profiling, chemoproteomics, and best-in-class molecular biology techniques. By these means I will be able to perform: 1) lead optimizationof OX01914 to identify a safe, stable, and potent lead candidate for preclinical studies; 2) perform chemoproteomic-based target engagementstudies to confirm its modulation of mitochondrial ATP synthase peripheral-stalk subunit b (ATP5F1) protein, previously identified as OX01914potential target; 3) investigate the molecular mechanisms leading to utrophin regulation. To date, DMD is still without a cure. Upregulating utrophin,an endogenous safe dystrophin paralogue, represents one of the most valuable strategies to generate a disease-modifying and accessible therapyfor DMD, applicable to all patients regardless of genetic mutations. On these premises, this MSCA project has the following objectives: WorkPackage 1. To rapidly generate an extended chemical library of OX01914 synthetic analogues using multicomponent reactions. Work Package 2.To identify a safe and metabolically stable lead candidate by in vitro activity assessment and ADME-T profiling of the new generated library. WorkPackage 3. To confirm ATP5F1 target engagement of DMD library using chemoproteomic approaches. Work Package 4. To provide newknowledge on how utrophin is modulated by nearing down biological factors involved, through RNA sequencing and proteome profiling, andinvestigating specific pathways using molecular biology techniques.