The present study describes an alkaline waterethanol mediated series of combinatorial synthesis of 2-amino-4-phenyl-5-H-indeno[1,2d]pyrimidine-5-one derivatives through sequential multicomponent reaction of 1,3indandione, aromatic aldehydes, and guanidine hydrochloride along with their anticancer evaluation. The effect of sequential addition of the components in the configuration of the desired product has been studied by UVvisible absorption spectroscopy. The synthetic method obeys most of the green chemistry principles in regard to high atom economy and greener, nontoxic, and noncarcinogenic solvent system (waterethanol). The selected synthesized compounds have been screened against the human breast cancer cell line MCF7, human colon cancer cell line HT29, and normal viro monkey cell line, out of which 2-amino-4-(4-methoxyphenyl)-5H-indeno[1,2-d]pyrimidin-5-one showed significant potency toward human breast cancer cell line (MCF7).