Mukherjee Avinaba, Adhikari Nilanjan, Jha Tarun
Natural Science Laboratory, Division of Medicinal and Pharmaceutical Chemistry, Department of Pharmaceutical Technology, P.O. Box 17020, Jadavpur University, Kolkata 700032, India.
Natural Science Laboratory, Division of Medicinal and Pharmaceutical Chemistry, Department of Pharmaceutical Technology, P.O. Box 17020, Jadavpur University, Kolkata 700032, India.
Eur J Med Chem. 2017 Dec 1;141:37-50. doi: 10.1016/j.ejmech.2017.09.052. Epub 2017 Sep 27.
Depending on our previous observations, some compounds of pentanoic acid were designed and synthesized. Characterization of the synthesized compounds was done by mass, NMR and IR spectroscopy as well as elemental analysis. Among the synthesized molecules, (2S)-5-oxo-2-[(nitrobenzene-4-yl sulfonyl) amino]-5-(pentylamino) pentanoic acid (Cpd 11) was found as a lead and potent inhibitor of matrix metalloproteinase-2 (MMP-2). Molecular modeling and enzyme inhibition studies were done to confirm the interaction or inhibitory potential of this compound. Thereafter, the biological screening was done through cytotoxicity, anti-invasion and apoptosis-related assays. Docking analysis revealed that Cpd 11 interacted with the target molecule MMP-2 and with MMP-9. However, enzyme inhibition assay showed 3-fold MMP-2 inhibition compared to MMP-9. Cytotoxicity assay showed the inhibitory potential of Cpd 11 against K562 cell line having IC value of 17.9 ± 0.01 μM after 48 h of incubation. The cell death was apoptotic in nature as revealed from the annexin V and sub-G1 cell cycle arrest assay. Besides this, Cpd 11 also exhibited dose dependent anti-invasive activity into K562 cell line. On the other hand, flow cytometry and western blot data revealed Cpd 11 induced downregulation of MMP-2 in K562 cell line after 48 h of incubation that might be linked with the anti-invasive and apoptotic activity furthermore. Therefore, the overall results validated each method and make this molecule as a potent MMP-2 inhibitor that blocked the invasion and could bring apoptosis at later stages in K562 cells sparing the normal ones.
根据我们之前的观察结果，设计并合成了一些戊酸化合物。通过质谱、核磁共振和红外光谱以及元素分析对合成的化合物进行了表征。在合成的分子中，发现(2S)-5-氧代-2-[(硝基苯-4-基磺酰基)氨基]-5-(戊基氨基)戊酸（化合物11）是基质金属蛋白酶-2（MMP-2）的先导且强效抑制剂。进行了分子建模和酶抑制研究以确认该化合物的相互作用或抑制潜力。此后，通过细胞毒性、抗侵袭和凋亡相关试验进行了生物学筛选。对接分析表明化合物11与靶分子MMP-2和MMP-9相互作用。然而，酶抑制试验显示与MMP-9相比，MMP-2的抑制作用提高了3倍。细胞毒性试验显示化合物11对K562细胞系具有抑制潜力，孵育48小时后IC值为17.9±0.01μM。膜联蛋白V和亚G1期细胞周期阻滞试验表明细胞死亡本质上是凋亡性的。除此之外，化合物11对K562细胞系也表现出剂量依赖性的抗侵袭活性。另一方面，流式细胞术和蛋白质印迹数据显示，孵育48小时后，化合物11在K562细胞系中诱导MMP-2下调，这可能进一步与抗侵袭和凋亡活性相关。因此，总体结果验证了每种方法，并使该分子成为一种强效的MMP-2抑制剂，可阻断K562细胞的侵袭，并在后期引发凋亡，同时使正常细胞不受影响。
