Al Fayi Majed Saad, Alshyarba Mishari
Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Khalid University, Abha 31982, Saudi Arabia.
Department of Surgery, College of Medicine, King Khalid University, Abha 31982, Saudi Arabia.
Curr Issues Mol Biol. 2025 Jun 17;47(6):463. doi: 10.3390/cimb47060463.
Tankyrase (TNKS1) regulates the WNT/β-catenin pathway, while CDK8 is a transcriptional regulator overexpressed in renal cell carcinoma (RCC). This study aims to identify novel dual inhibitors of tankyrase and Cyclin-dependent kinase 8 (CDK8), utilizing bioinformatics and in vitro methods and to assess their efficiency in renal cancer cells.
To identify leads, the ChemBridge library was screening using high-throughput virtual screening (HTVS), which was followed by protein-ligand interaction analysis, Molecular Dynamics (MD) simulation, and Gibbs binding free energy estimation. A-498, Caki-1, and HK-2 cells were employed to validate in vitro efficacy.
TCS9725 was discovered by HTVS with binding affinities of -8.1 kcal/mol and -8.2 kcal/mol for TNKS1 and CDK8, respectively. TCS9725 had robust binding interactions with root mean square deviation values of 0.00 nm. The ΔG binding estimate was -27.45 for TNKS1 and -27.88 for CDK8, respectively. ADME predictions favored specific small-molecule inhibition profiles. TCS9725 reduced TNKS1 and CDK8 activities with IC50s of 243 nM and 403.6 nM, respectively. The compound efficiently inhibited the growth of A-498 and Caki-1 cells with GI50 values of 385.9 nM and 243.6 nM, respectively, with high selectivity compared to the non-cancerous kidney cells. TCS9725 decreased STAT1 and β-catenin positivity in A-498 and Caki-1 cells. The compound induced apoptosis and reduced TGFβ-stimulated trans-endothelial migration and p-smad2/3 signaling in both RCC cells.
This work provides valuable insights into the therapeutic potential of TCS9725, a dual inhibitor of TNKS1 and CDK8. Further developments of this molecule could lead to new and effective treatments for this devastating disease.
端锚聚合酶（TNKS1）调节WNT/β-连环蛋白信号通路，而细胞周期蛋白依赖性激酶8（CDK8）是一种在肾细胞癌（RCC）中过表达的转录调节因子。本研究旨在利用生物信息学和体外方法鉴定端锚聚合酶和细胞周期蛋白依赖性激酶8（CDK8）的新型双重抑制剂，并评估它们在肾癌细胞中的效能。
为了鉴定先导化合物，使用高通量虚拟筛选（HTVS）对ChemBridge文库进行筛选，随后进行蛋白质-配体相互作用分析、分子动力学（MD）模拟和吉布斯结合自由能估计。采用A-498、Caki-1和HK-2细胞验证体外效能。
通过高通量虚拟筛选发现了TCS9725，其对TNKS1和CDK8的结合亲和力分别为-8.1 kcal/mol和-8.2 kcal/mol。TCS9725具有强大的结合相互作用，均方根偏差值为0.00 nm。TNKS1和CDK8的结合自由能估计值分别为-27.45和-27.88。药物代谢动力学预测支持特定的小分子抑制谱。TCS9725降低TNKS1和CDK8的活性，IC50分别为243 nM和403.6 nM。该化合物有效抑制A-498和Caki-1细胞的生长，GI50值分别为385.9 nM和243.6 nM，与非癌性肾细胞相比具有高选择性。TCS9725降低A-498和Caki-1细胞中STAT1和β-连环蛋白的阳性表达。该化合物诱导细胞凋亡，并减少两种肾癌细胞中TGFβ刺激的跨内皮迁移和p-smad2/3信号传导。
本研究为TNKS1和CDK8的双重抑制剂TCS9725的治疗潜力提供了有价值的见解。该分子的进一步开发可能会为这种毁灭性疾病带来新的有效治疗方法。
