A dual-tail approach was applied to the design of a novel series of 2-thiopyrimidine–benzenesulfonamides as carbonic anhydrase (CA) inhibitors. The design strategy is based on the hybridization between a benzenesulfonamide moiety as Zn²⁺ binding group and 2,4-disubstituted thiopyridimidine as a tail. Among the synthesized compounds, 14h displayed the highest potency (K_i = 1.72 nM) and selectivity for CA II over the isoforms CA IX and CA XII with selectivity indexes of 50 and 5.26, respectively. Meanwhile, compounds 14a and 14l displayed a potent inhibitory activity against CA IX (K_i = 7.4 and 7.0 nM, respectively) compared with the reference drug acetazolamide (AAZ) (K_i = 25 nM), and compound 14l showed higher potency (K_i = 4.67 nM) than AAZ (K_i = 5.7 nM) against the tumor-associated isoform CA XII. Evaluation of the antiproliferative activity in NCI single-dose testing of selected hybrids revealed a pronounced potency of the selective CA II inhibitor 14h against most of the tested NCI cancer cell lines. Moreover, compound 14h demonstrated an IC₅₀ values ranging from 2.40 to 4.50 μM against MCF-7, T-47D, MDA-MB-231, HCT-116, HT29 and SW-620. These results demonstrate that CA II inhibition can be an alternative therapeutic target for cancer treatment. A cell cycle analysis of MCF-7 and MDA-MB-231 showed that treatment with 14h arrested both cell lines at the G2/M phase with significant accumulation of cells in the pre-G1 phase. Moreover, compound 14h showed a noticeable induction of late apoptosis and necrotic cell death of both cell lines compared with untreated cells as a control. A molecular docking study suggested that the sulfonamide moiety accommodates deeply in the CA active site and interacts with the Zn²⁺ ion while the dual-tail extension interacts with the surrounding amino acids via several hydrophilic and hydrophobic interactions, which affects the potency and selectivity of the hybrids.

采用双尾方法设计了一系列新型 2-硫代嘧啶-苯磺酰胺作为碳酸酐酶 (CA) 抑制剂。设计策略基于苯磺酰胺部分作为 Zn ²⁺结合基团和 2,4-二取代硫代嘧啶作为尾部之间的杂交。在合成的化合物中，14h对 CA II的效力（K _i  = 1.72 nM）和选择性高于同工型 CA IX 和 CA XII，选择性指数分别为 50 和 5.26。同时，化合物14a和14l对 CA IX ( K _i = 7.4 和 7.0 nM，分别与参考药物乙酰唑胺 ( AAZ ) ( K _i  = 25 nM) 相比，化合物14l显示出比AAZ ( K _i  = 5.7 nM)更高的抗肿瘤效力 ( K _i  = 4.67 nM) -相关的异构体 CA XII。对选定杂种的 NCI 单剂量测试中抗增殖活性的评估揭示了选择性 CA II 抑制剂 14 小时对大多数测试的 NCI 癌细胞系的显着效力。此外，化合物14h显示出 IC ₅₀对 MCF-7、T-47D、MDA-MB-231、HCT-116、HT29 和 SW-620 的值范围为 2.40 至 4.50 μM。这些结果表明，CA II 抑制可以成为癌症治疗的替代治疗靶点。MCF-7 和 MDA-MB-231 的细胞周期分析表明，14 小时的处理使两种细胞系都处于 G2/M 期，细胞在 G1 前的显着积累。此外，与作为对照的未处理细胞相比，化合物14h显着诱导了两种细胞系的晚期凋亡和坏死细胞死亡。分子对接研究表明，磺胺部分深深地容纳在 CA 活性位点并与 Zn ^{2+相互作用}离子，而双尾延伸通过几种亲水和疏水相互作用与周围的氨基酸相互作用，从而影响杂合体的效力和选择性。