Some novel N-nicotinonitrile derivatives 3-14 have been synthesized starting with compound 1. The key step of this work is the coupling between compound 1 and activated sugars to afford the corresponding cyclic nucleosides 3-6. Moreover, the cytotoxicity and in vitro anticancer evaluation of the prepared compounds have also been assessed against breast MCF-7 cancer, liver HepG2 cancer and lung A549 carcinoma cell lines with investigation the effect of the synthesized compounds on the expression of urokinase plasminogen activator (uPA). The results revealed that, although all the compounds showed no anticancer activity against A549 cells without showing any effect on the expression of uPA, the tested compounds exhibited remarkable cytotoxicity activity against MCF-7 and HepG2 cell lines. Among the tested compounds, compounds 11 and 12 revealed promising anticancer activity compared to the activity of the commonly used anticancer drug, doxorubicin with inhibiting the expression of uPA.