Abstract：
OBJECTIVE: Exploring the efficacy of β-carboline-based molecular inhibitors in targeting microtubules for the development of novel anticancer therapeutics.
METHODS: We synthesized a series of 1-Aryl-N-substituted-β-carboline-3-carboxamide compounds and evaluated their cytotoxicity against human lung carcinoma (A549) cells using the MTT assay. Normal lung fibroblast cells (WI-38) were used to assess compound selectivity. The mechanism of action of MJ-211 was elucidated through Western blot analysis of key pro-apoptotic and cell cycle regulatory proteins. Additionally, the inhibitory effect of MJ-211 on multicellular 3D spheroid growth of A549 cells was evaluated.
RESULTS: Lead compound MJ-211 exhibited remarkable cytotoxicity against A549 cells with an IC50 of 4.075 μM at 24 h treatment and IC50 of 1.7 nM after 72 h of treatment, while demonstrating selectivity towards normal WI-38 cells. MJ-211 activated pro-apoptotic factors Bim and p53, and suppressed Cyclin B1, Phospho HSP 27, BubR1, Mad 2, ERK1/2, and NF-κB, indicating its potent antimitotic and pro-apoptotic effects. MJ-211 significantly suppressed the migration of cells and inhibited the growth of A549 cell-derived multicellular 3D spheroids, highlighting its efficacy in a more physiologically relevant model.
CONCLUSIONS: Cytotoxic effect of MJ-211 against cancer cells, selectivity towards normal cells, and ability to modulate key regulatory proteins involved in apoptosis and cell cycle progression underscore its potential as a promising template for further anticancer lead optimization. Moreover, the inhibitory effect of MJ-211 on multicellular spheroid growth suggests its efficacy in combating tumor heterogeneity and resistance mechanisms, thereby offering a promising avenue for future anticancer drug development.
METHODS: We synthesized a series of 1-Aryl-N-substituted-β-carboline-3-carboxamide compounds and evaluated their cytotoxicity against human lung carcinoma (A549) cells using the MTT assay. Normal lung fibroblast cells (WI-38) were used to assess compound selectivity. The mechanism of action of MJ-211 was elucidated through Western blot analysis of key pro-apoptotic and cell cycle regulatory proteins. Additionally, the inhibitory effect of MJ-211 on multicellular 3D spheroid growth of A549 cells was evaluated.
RESULTS: Lead compound MJ-211 exhibited remarkable cytotoxicity against A549 cells with an IC50 of 4.075 μM at 24 h treatment and IC50 of 1.7 nM after 72 h of treatment, while demonstrating selectivity towards normal WI-38 cells. MJ-211 activated pro-apoptotic factors Bim and p53, and suppressed Cyclin B1, Phospho HSP 27, BubR1, Mad 2, ERK1/2, and NF-κB, indicating its potent antimitotic and pro-apoptotic effects. MJ-211 significantly suppressed the migration of cells and inhibited the growth of A549 cell-derived multicellular 3D spheroids, highlighting its efficacy in a more physiologically relevant model.
CONCLUSIONS: Cytotoxic effect of MJ-211 against cancer cells, selectivity towards normal cells, and ability to modulate key regulatory proteins involved in apoptosis and cell cycle progression underscore its potential as a promising template for further anticancer lead optimization. Moreover, the inhibitory effect of MJ-211 on multicellular spheroid growth suggests its efficacy in combating tumor heterogeneity and resistance mechanisms, thereby offering a promising avenue for future anticancer drug development.
摘要：
目的：探索基于β-咔啉的分子抑制剂靶向微管的功效，以开发新的抗癌疗法。
方法：我们合成了一系列1-芳基-N-取代-β-咔啉-3-甲酰胺化合物，并使用MTT法评估了它们对人肺癌（A549）细胞的细胞毒性。正常肺成纤维细胞(WI-38)用于评估化合物选择性。通过蛋白质印迹分析关键的促凋亡和细胞周期调节蛋白，阐明了MJ-211的作用机制。此外,评价MJ-211对A549细胞多细胞三维球状体生长的抑制作用。
结果：先导化合物MJ-211对A549细胞具有显著的细胞毒性，24小时处理后IC50为4.075μM，72小时处理后IC50为1.7nM，同时显示对正常WI-38细胞的选择性。MJ-211激活促凋亡因子Bim和p53，并抑制细胞周期蛋白B1，磷酸化HSP27，BubR1，Mad2，ERK1/2和NF-κB，表明其有效的抗有丝分裂和促凋亡作用。MJ-211显著抑制细胞迁移,抑制A549细胞来源的多细胞3D球体生长,在更生理相关的模型中强调其功效。
结论：MJ-211对癌细胞的细胞毒性作用，对正常细胞的选择性，和调节参与细胞凋亡和细胞周期进程的关键调节蛋白的能力强调了其作为进一步抗癌线索优化的有希望的模板的潜力。此外，MJ-211对多细胞球体生长的抑制作用表明其在对抗肿瘤异质性和耐药机制方面的功效，从而为未来的抗癌药物开发提供了有希望的途径。
方法：我们合成了一系列1-芳基-N-取代-β-咔啉-3-甲酰胺化合物，并使用MTT法评估了它们对人肺癌（A549）细胞的细胞毒性。正常肺成纤维细胞(WI-38)用于评估化合物选择性。通过蛋白质印迹分析关键的促凋亡和细胞周期调节蛋白，阐明了MJ-211的作用机制。此外,评价MJ-211对A549细胞多细胞三维球状体生长的抑制作用。
结果：先导化合物MJ-211对A549细胞具有显著的细胞毒性，24小时处理后IC50为4.075μM，72小时处理后IC50为1.7nM，同时显示对正常WI-38细胞的选择性。MJ-211激活促凋亡因子Bim和p53，并抑制细胞周期蛋白B1，磷酸化HSP27，BubR1，Mad2，ERK1/2和NF-κB，表明其有效的抗有丝分裂和促凋亡作用。MJ-211显著抑制细胞迁移,抑制A549细胞来源的多细胞3D球体生长,在更生理相关的模型中强调其功效。
结论：MJ-211对癌细胞的细胞毒性作用，对正常细胞的选择性，和调节参与细胞凋亡和细胞周期进程的关键调节蛋白的能力强调了其作为进一步抗癌线索优化的有希望的模板的潜力。此外，MJ-211对多细胞球体生长的抑制作用表明其在对抗肿瘤异质性和耐药机制方面的功效，从而为未来的抗癌药物开发提供了有希望的途径。
