DataSheet3_Design, Synthesis, and Apoptosis-Promoting Effect Evaluation of Rhopaladins’ Analog 4-Arylidene-5-Oxopyrrolidine Derivatives.docx

Marine alkaloids have novel structures and antitumor activities. Therefore, we synthesized rhopaladins’ analogs from marine alkaloids rhopaladins A-D and modified their structures to synthesize 4-benzylidene-5-pyrrolidone derivatives. Among the compounds, (2E, 4E)-4-(4-chlorobenzylidene)-2-(4-chlorostyryl)-N-cyclohexyl-1-(4-fluorophenyl)-5-oxopyrrolidine-2-carboxamide (RPDPRH) has high efficiency and less hepatotoxicity, with IC50 values of 4.66, 6.42, 17.66, 15.2, 12.36, 22.4, and 243.2 μM in vitro anti-proliferative activity testing against cervical cancer C-33A, CaSki, SiHa, and HeLa cells, human hepatocarcinoma HepG2 and 7402 cells, and human normal liver LO2 cells, respectively. In particular, RPDPRH has similar activity to cisplatin on human hepatocarcinoma cells, and cisplatin served as a positive control in our study. Next, the apoptosis of HepG2 and 7402 cells induced by RPDPRH at different concentrations was detected by Annexin V/PI flow cytometry. Moreover, the expression of apoptotic proteins was detected by Western blot analysis. Finally, the results showed that RPDPRH could induce apoptosis of hepatocarcinoma cells by regulating Bax and Bcl-2 expressions. In summary, our results indicate that RPDPRH has the potential to serve as an antitumor agent and plays a significant role in future studies.

海洋生物碱具有新颖的结构和抗癌活性。因此，我们从海洋生物碱 rhopaladins A-D 中合成了 rhopaladins 的类似物，并对其结构进行了修饰，以合成 4-苄基甲酰基-5-吡咯烷酮衍生物。在这些化合物中，(2E,4E)-4-(4-氯苄基甲酰基)-2-(4-氯苯乙烯基)-N-环己基-1-(4-氟苯基)-5-氧吡咯烷-2-羧酰胺（RPDPRH）表现出高效率和低肝毒性，其在体外抗增殖活性测试中对宫颈癌 C-33A、CaSki、SiHa 和 HeLa 细胞、人肝细胞癌 HepG2 和 7402 细胞以及人正常肝细胞 LO2 细胞的 IC50 值分别为 4.66、6.42、17.66、15.2、12.36、22.4 和 243.2 μM。特别是，RPDPRH 在人肝细胞癌细胞上的活性与顺铂相似，顺铂在本研究中作为阳性对照。随后，通过 Annexin V/PI 流式细胞术检测了不同浓度的 RPDPRH 诱导 HepG2 和 7402 细胞凋亡的情况。此外，通过蛋白质印迹分析检测了凋亡蛋白的表达。最终结果显示，RPDPRH 通过调节 Bax 和 Bcl-2 的表达，能够诱导肝细胞癌细胞的凋亡。总之，我们的研究结果表明，RPDPRH 具有作为抗癌剂的潜力，并在未来的研究中发挥着重要作用。