Three series of indolinone-based sulfonamides (3a–f, 6a–f and 9a–f) were in vitro evaluated as inhibitors of the tumor-associated carbonic anhydrase (CA, EC 4.2.1.1) isoforms hCA IX and XII, using a stopped-flow CO₂ hydrase assay. All the investigated sulfonamides displayed single- or double-digit nanomolar inhibitory activities towards both hCA IX (K_Is: 6.2–64.8 nM) and XII (K_Is: 7.1–55.6 nM) isoforms. All sulfonamides (3a–f, 6a–f and 9a–f) were in vitro examined for their potential anticancer activity against colorectal cancer HCT-116 and breast cancer MCF-7 cell lines. Sulfonamide 9e was found to be the most potent counterpart against HCT-116 (IC₅₀ = 3.67 ± 0.33 µM). Sulfonamide 9e displayed good selectivity profile for inhibition of the tumor-associated isoforms (CAs IX & XII) over the off-target cytosolic CAs I and II. 9e was screened for cell cycle disturbance and apoptosis induction in HCT-116 cells. It was found to persuade cell cycle arrest at G₂-M stage as well as alter the Sub-G₁ phase. Also, 9e induced the intrinsic apoptotic mitochondrial pathway in HCT-116 cells via down-regulation of the anti-apoptotic protein Bcl-2 level with concurrent boosting the pro-apoptotic Bax, caspase-9, caspase-3, cytochrome C and p53 levels.

体外评估了三系列基于吲哚酮的磺酰胺（3a–f，6a–f和9a–f）作为与肿瘤相关的碳酸酐酶（CA，EC 4.2.1.1）同工型hCA IX和XII的抑制剂，使用了终止剂流CO ₂酶测定。所有研究的磺酰胺类药物均对hCA IX（K _I s：6.2-64.8 nM）和XII（K _I s：7.1-55.6 nM）同种型均显示出一位或两位数纳摩尔抑制活性。所有磺酰胺类药物（3a–f，6a–f和9a–f）均在体外检查了它们对结肠直肠癌HCT-116和乳腺癌MCF-7细胞系的潜在抗癌活性。发现磺酰胺9e是对抗HCT-116的最有效对应物（IC ₅₀  = 3.67±0.33 µM）。磺酰胺9e表现出良好的选择性，可抑制脱靶胞质CAs I和II抑制肿瘤相关同工型（CAs IX和XII）。筛选9e在HCT-116细胞中的细胞周期干扰和凋亡诱导。已发现其能说服细胞周期停滞在G ₂ -M期以及改变Sub-G ₁期。另外，图9e诱导HCT-116细胞中的内在凋亡线粒体途径经由下调抗凋亡蛋白Bcl-2的水平，同时增强促凋亡的Bax，caspase-9，caspase-3，细胞色素C和p53的水平。