腺苷酸环化酶抑制剂体外筛选模型的建立及应用

腺苷酸环化酶(adenylate cyclase,AC)通过调节环磷酸腺苷(cyclic adenosine cyclophosphate,cAMP)的合成从而调控细胞相关功能.在卵巢中,cAMP调控卵母细胞的发育及卵子的成熟,cAMP含量变化影响下游芳香化酶的表达,进而影响雌激素的生物合成.为开发靶向AC的候选药物,...

Establishment and application of a screening model of adenylate cyclase inhibitor in vitro

Adenylate cyclase(AC) affects cellular functions by regulating the synthesis of cyclic adenosine monophosphate(cAMP), which regulates oocyte development and follicle maturation in the ovary. Changes in cAMP content modulates the expression of aromatase and the biosynthesis of estrogen. Therefore, inhibition of AC by small molecules may provide new therapeutic means for the treatment of estrogen-related diseases. In order to develop candidate drugs targeting AC, an Escherichia coli system was used to clone and express soluble ⅠC1-ⅡC2 recombinant fusion proteins based on the two active regions of AC1 and AC2. The catalytic activity of ⅠC1-ⅡC2 was evaluated using an ATP detection kit. A chemical library consisting of 2 861 compounds was screened for inhibitors of ⅠC1-ⅡC2. The results showed that the compound neratinib significantly inhibited ⅠC1-ⅡC2 activity in a concentration-dependent manner, with an IC50 value of 10.2 ± 0.1522 μmol/L. Furthermore, neratinib significantly inhibited the expression of aromatase and the biosynthesis of estrogen in human ovarian granulose-like KGN cells. This study established a high-throughput screening model of ⅠC1-ⅡC2 protein to find a highly active AC inhibitor in vitro, providing a good experimental foundation for the study of infertility treatment.