A series of pyrazolo[3,4-d]pyrimidin-4-one scaffold were designed and synthesized as novel CDK2 inhibitors. By analyzing the common motifs of various known inhibitors, the designed compounds 1 were virtually screen for their inhibitory activity by docking into the active pocket of CDK2. The influence of different substitutes on the docking results was investigated. A total of 15 pyrazolo[3,4-d]pyrimidin-4-ones 1 were synthesized by Paal-Knorr reaction, pyrimidine ring closure, bromination, Suzuki coupling reaction, amide formation and Knoevenagel condensation. The Cell Counting Kit-8 (CCK-8) was used to evaluate the inhibitory activity of pyrazolo[3,4-d]pyrimidin-4-ones 1 in the breast cancer cell line MCF-7 in vitro using Etoposide as a reference control substance. The screening results demonstrated that the designed compounds have significant antiproliferative activity, and compounds 1e and 1j were the most active compounds with IC₅₀ values of 10.79 μM and 10.88 μM, respectively, being better than that of Etoposide (IC₅₀ = 18.75 μM). The enzyme inhibition assay was carried out against CDK2, the results indicated that the compounds 1e and 1j significantly inhibited CDK2 with IC₅₀ values of 1.71 μM and 1.60 μM.

一系列吡唑并[3,4- d ]pyrimidin-4-one 支架被设计并合成为新型CDK2 抑制剂。通过分析各种已知抑制剂的共同基序，设计的化合物1通过与 CDK2 的活性口袋对接来虚拟筛选其抑制活性。考察了不同替代品对对接结果的影响。通过Paal-Knorr反应、嘧啶闭环、溴化、Suzuki偶联反应、酰胺形成和Knoevenagel缩合反应共合成了15个吡唑并[3,4- d ]pyrimidin-4-ones 1 。Cell Counting Kit-8 (CCK-8) 用于评估 pyrazolo[3,4- d ]pyrimidin-4-ones 1的抑制活性在乳腺癌细胞系 MCF-7中使用依托泊苷作为参考对照物质。筛选结果表明，所设计的化合物具有显着的抗增殖活性，化合物1e和1j是活性最强的化合物，IC ₅₀值分别为10.79 μM和10.88 μM，优于依托泊苷（IC ₅₀  = 18.75 μM）。对CDK2进行酶抑制试验，结果表明化合物1e和1j显着抑制CDK2，IC ₅₀值分别为1.71 μM和1.60 μM。