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

Look-Up旨在开发一种新近发现的碳肼-嘧啶促性腺激素上调因子，即OX01914，以产生一种治疗Duchenne肌营养不良症(DMD)的最佳小分子药物。Duchenne肌营养不良症是一种罕见的遗传性肌肉萎缩致命疾病。为此，我和罗素教授依靠药物化学、体外高通量(HT)ADME-T图谱、化学蛋白质组学和一流的分子生物学技术，设计了一种多学科和互补的体外方法。通过这些手段，我将能够执行：1)OX01914的先导优化，以确定安全、稳定和有效的临床前研究的先导候选者；2)进行基于化学蛋白质组的靶参与研究，以确认其对线粒体ATP合成酶外周茎亚单位b(ATP5F1)蛋白的调节，该蛋白以前被确定为OX01914的潜在靶点；3)研究导致促营养素调节的分子机制。到目前为止，DMD仍然没有治愈的方法。上调utroin是一种内源性安全的dystrophin类似物，是产生一种可改变疾病和可获得的治疗DMD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.利用多组分反应快速生成OX01914合成类似物的扩展化学库。工作包2.通过对新生成的文库进行体外活性评估和ADME-T图谱分析，确定安全和代谢稳定的候选先导化合物。用化学蛋白质组学方法确定DMD文库的ATP5F1靶结合。工作包4.通过RNA测序和蛋白质组图谱，以及利用分子生物学技术研究特定的途径，提供关于如何通过接近所涉及的生物因素来调节促营养素的新知识。

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.