Bisphosphorylation of hydroquinone and 4,4′-(propane-2,2-diyl)diphenol was performed for the first time in ethyl acetate. The bisphosphorylated products were used in molecular assembly of unsymmetrical phosphorus(III)-containing macrocycles consisting of hydroquinone and 2,2-bis( p-hydroxyphenyl)propane or resorcinol fragments. The macrocyclic compound based on hydroquinone and 2,2-bis( p-hydroxyphenyl)-propane was subjected to oxidation and sulfurization to obtain the corresponding phosphorus(V) derivatives. Its biological activity was also studied.