Carbonic anhydrase-II (CA-II) catalyzes reversible hydration of carbon dioxide leading to the formation of bicarbonate and a proton. CA-II inhibitors act as biomarkers and therapeutic targets for various diseases such as epilepsy, glaucoma, leukemia, and cystic fibrosis. In the present work, a number of thiourea and benzylidene derivatives were evaluated as CA-II inhibitors. For this, detection of CA-II inhibition was tested through high throughput biochemical mechanism-based assay. Results indicated that thiourea derivatives exhibit remarkable inhibition activity with IC₅₀ values ranging from 1.90 ± 1.30 to 25.90 ± 2.05 µM, when compared with the standard inhibitor acetazolamide IC₅₀ = 0.13 ± 0.05. On the other hand, bezylidene derivatives showed weak inhibition potential with IC₅₀ values in the range of 52.68 ± 0.47 to 348.57 ± 3.32 µM when compared with the thiourea derivatives; the Ez-Fit enzyme kinetics software was used for IC₅₀ calculations from the percentage inhibition. In addition, 3T3 normal cell line was employed to determine the toxicity profile of identified inhibitors of CA-II. In silico modeling was performed to decipher the molecular nature of thiourea and benzylidene derivatives with key residues of carbonic anhydrase; results were in agreement with experiments related to inhibition of carbonic anhydrase.

碳酸酐酶-II（CA-II）催化二氧化碳的可逆水合作用，导致形成碳酸氢盐和质子。CA-II抑制剂可作为癫痫，青光眼，白血病和囊性纤维化等各种疾病的生物标志物和治疗靶标。在目前的工作中，许多硫脲和亚苄基衍生物被评估为CA-II抑制剂。为此，通过基于高通量生化机制的分析测试了对CA-II抑制的检测。结果表明，与标准抑制剂乙酰唑胺IC ₅₀  = 0.13±0.05相比，硫脲衍生物表现出显着的抑制活性，IC ₅₀值为1.90±1.30至25.90±2.05 µM 。另一方面，苄叉衍生物对IC的抑制能力较弱与硫脲衍生物相比，_50个值在52.68±0.47至348.57±3.32 µM范围内；Ez-Fit酶动力学软件用于根据抑制百分比计算IC ₅₀。另外，使用3T3正常细胞系来确定已鉴定的CA-II抑制剂的毒性谱。进行了计算机模拟以破译带有碳酸酐酶关键残基的硫脲和亚苄基衍生物的分子性质。结果与有关抑制碳酸酐酶的实验一致。