The presented work is dedicated to the synthesis of cyclohexane fragment containing aminomethoxy derivatives based on the three-component Mannich reaction involving cyclohexanole, formaldehyde, and secondary aliphatic amines (diethyl, dipropyl, dibutyl, dihexyl), as well as the study of their properties. The reaction is carried out by mixing the starting materials in equimolar amounts in a benzene medium at 78-80°C for 4–5 hours. During the reaction, CuCl was employed as a catalyst, leading to a high yield of the synthesized products. The yields of the compounds 83–91%. The synthesized new aminomethoxy derivatives’ physicochemical properties were determined. These substances are yellowish liquids with a distinctive odor. They are insoluble in water but dissolve in organic solvents (such as ethanol, acetone, benzene, chloroform, CCl4 and others). The composition and structures of N,N-diethylaminomethoxycyclohexane, N,N-dipropylaminomethoxycyclohexane, N,N-dibutylaminomethoxycyclohexane and N,N-dihexylaminomethoxy cyclohexane were confirmed by elemental analysis, IR and NMR spectroscopy methods. As a result of the synthesis of new aminomethoxy derivatives containing a cyclohexane fragment, compounds with high biological activity have been obtained. The antimicrobial activity of the synthesized compounds was studied using the serial dilution method against various microorganisms. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the synthesized aminomethoxy derivatives of cyclohexanol were determined against bacteria and fungi (S.aureus, E.coli, C.albicans).