Discovery Of A Small-molecule Inhibitor Of CPS1 And Anti-colorectal Cancer Mechanism

Carbamoyl phosphate synthase 1（CPS1）is involved in the first step of the urea cycle is a key rate-limiting enzyme in the urea cycle.CPS1 maintains ammonia levels within cells by participating in processes such as the urea cycle.Also,it plays a role in ammonia detoxification,as well as pyrimidine and arginase synthesis.Researchers found CPS1 expression to be high in a wide range of tumor types,including colorectal cancer,lung cancer,esophageal cancer,prostate cancer,glioblastoma,bladder cancer,endometrial cancer,and its overexpression correlated with a poor prognosis of tumor cells.Therefore,small molecules targeted to inhibit the function of CPS1 in tumors may provide therapeutic benefits for cancer patients who overexpress CPS1.In this project,in vitro studies on the process of CPS1 found that overexpression of CPS1 can enhance the migration ability of colorectal cancer cells HCT15.In addition,8 small-molecule compounds were screened from the Drug Bank and SPECS libraries as CPS1 candidate small-molecule inhibitors based on a high-throughput virtual screening method.In the colorectal cancer cell line（HCT15,HCT116）,the antiproliferative activity of eight small molecule candidate inhibitors was evaluated.It was found that compound 3 had good antiproliferative activity(HCT15,IC₅₀,8.476μM,HCT116,IC₅₀,13.5μM)of colorectal cancer HCT15 and HCT116 cell lines as potential small molecule inhibitors targeting CPS1.Molecular docking studies have shown that compound 3 interacts with crucial amino acids at the Allomorphic sites of CPS1 by forming hydrogen bonds and hydrophobic interactions.In vitro studies found that compound 3 can inhibit the proliferation of colorectal cancer cell lines and lung cancer cell lines and can significantly weaken the migration ability of colorectal cancer cell lines and lung cancer cells.We also found that compound 3 can block the S-stage progression of colorectal cancer cells and induce apoptosis.It verifies in vitro activity with high-throughput screening and preliminary research on the anti-tumor mechanism of action.It was determined that compound 3 is a potential allosteric inhibitor targeting CPS1,which provides both the experimental basis and theoretical basis for the development of targeted intervention small molecule therapeutic drugs.At the same time,compound 3 can be used to evaluate further the therapeutic potential of targeted inhibition of CPS1 in animal models and further optimize its activity and therapeutic potential with compound 3 as the parent nucleus,providing therapeutic benefits for patients with CPS1-related tumors.