Abohassan Mohammad, Shahrani Mesfer Mohammad Al, Alouda Sarah Khaled, Rajagopalan Prasanna
Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Khalid University, Abha, 61413, Saudi Arabia.
Oncol Res. 2025 Feb 28;33(3):675-685. doi: 10.32604/or.2024.056256. eCollection 2025.
The JAK2 mutation plays a crucial part in the pathogenesis of myeloproliferative neoplasms (MPN), which includes polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF) leading to aberrant proliferation and survival of hematopoietic cells. Alongside the challenges of drug resistance and side effects, identifying novel compounds that selectively target JAK2 could provide more effective and safer therapeutic options for patients with MPNs.
We employed computational approaches like high-throughput virtual screening, molecular dynamics simulations (MDS), and binding free energy calculations to identify inhibitors targeting wild and mutant JAK2 kinases. JAK2 positive HEL, wild type JAK2 positive TF-1, and non-cancerous Vero cells were used for validations.
SBLJ23 emerged as a top candidate inhibitor with specificity for JAK2. Protein-ligand interaction studies and MDS revealed stable interactions and binding of SBLJ23 over the simulation period, with Root Mean Square Deviation (RMSD) indicating consistent binding after 1t15ns. SBLJ23 displayed a half maximal inhibitory concentration (IC) value of 522.4 nM against the JAK2 enzyme. The compound exhibited inhibition of cell proliferation in HEL and TF-1 cells, with half maximal cell growth inhibitory concentration (GI) values of 2.51 and 15.87 µM, respectively. Moreover, SBLJ23 induced G/M cell cycle arrest in HEL cells to facilitate apoptosis in these cell lines. The compound significantly reduced the percentage of phospho JAK2 and phospho STAT3 in HEL cells.
High binding affinity, stable interaction profile, favorable binding free energy, and validations claim SBLJ23 as a potential lead compound against JAK2 and suggest further development and optimization towards clinical application in managing myeloproliferative neoplasms.
JAK2突变在骨髓增殖性肿瘤（MPN）的发病机制中起关键作用，MPN包括真性红细胞增多症（PV）、原发性血小板增多症（ET）和原发性骨髓纤维化（PMF），可导致造血细胞异常增殖和存活。除了耐药性和副作用的挑战外，鉴定选择性靶向JAK2的新型化合物可为MPN患者提供更有效、更安全的治疗选择。
我们采用高通量虚拟筛选、分子动力学模拟（MDS）和结合自由能计算等计算方法来鉴定靶向野生型和突变型JAK2激酶的抑制剂。使用JAK2阳性的HEL细胞、野生型JAK2阳性的TF-1细胞和非癌性Vero细胞进行验证。
SBLJ23成为对JAK2具有特异性的顶级候选抑制剂。蛋白质-配体相互作用研究和MDS显示，在模拟期间SBLJ23具有稳定的相互作用和结合，均方根偏差（RMSD）表明在15 ns后结合一致。SBLJ23对JAK2酶的半数最大抑制浓度（IC）值为522.4 nM。该化合物在HEL和TF-1细胞中均表现出对细胞增殖的抑制作用，其半数最大细胞生长抑制浓度（GI）值分别为2.51和15.87 μM。此外，SBLJ23诱导HEL细胞发生G/M期细胞周期阻滞，以促进这些细胞系的凋亡。该化合物显著降低了HEL细胞中磷酸化JAK2和磷酸化STAT3的百分比。
高结合亲和力、稳定的相互作用谱、良好的结合自由能以及验证结果表明SBLJ23是一种针对JAK2的潜在先导化合物，并建议进一步开发和优化以用于骨髓增殖性肿瘤管理的临床应用。
