AbstractBased on the wide range of pharmacological aspects related to organophosphates, a new type of chalcone‐phosphonate containing derivatives were synthesized by the reaction of dialkyl phosphite and substituted chalcones using anhydrous Mg(ClO4)2 at 80 °C under solvent‐free conditions. The resulting compounds were evaluated for anti‐proliferative activity in vitro against a panel of three human cancer cell lines: MCF7, HeLa, and A549 cell lines. Compound 3 l, 3 m, and 3 p exhibited anti‐proliferative activity against MCF7 and HeLa, with IC50 values of 1.12 μM, 1.63 μM, 1.09 μM and 1.29 μM, 1.44 μM, 2.40 μM respectively. The molecular docking study showed that the synthesized derivatives have good binding ability in the active site of the Vaccinia H1‐related (VHR) phosphatase (PDB: 3F81), PI3‐ kinase (PDB: 3R7Q), androgen receptor (PDB: 3V49) and VEGFR2 kinase (PDB: 3VHE). Finally, in silico pharmacokinetic (ADME) and toxicity studies revealed that compounds have the drug‐like feature for the favourable bioavailability. In conclusion, this work confirmed the potency of chalcone‐phosphonates derivatives for the further optimization of potential anti‐ proliferative agents.