Aim. Synthesis of a combinatorial library of substituted 3-sulfonyl-2-imino-1,2-dihydro-5H-dipyrido[1,2-a:2’,3’-d]pyrimidin-5-ones and evaluation of in vitro anticancer activity against a panel of 60 different human tumor cell lines derived from nine cancer types. Methods. Organic synthesis; biological tests; spectral methods; statistic methods. Results. An efficient protocol for the synthesis of new 3-sulfonyl-2-imino-1,2-dihydro-5H-dipyrido[1,2-a:2’,3’-d]pyrimidin-5-ones has been developed. In vitro screening of the anticancer activity showed that the obtaining compounds can effectively inhibit the growth of certain cancer cell lines. Conclusions. The synthesis of a library of 3‑sulfonyl-2-imino-1,2-dihydro-5H-dipyrido[1,2-a:2’,3’-d]pyrimidin-5-ones is described. The set of compounds (25 examples) inhibits the growth of certain renal cancer lines.