This study investigates the synthesis, antimicrobial activity, molecular docking, molecular dynamics simulation, and ADMET properties of mannopyranoside derivatives as antimicrobial agents. The research aimed to explore the potential of these compounds against various pathogens, including bacteria and fungi. The study involved the synthesis of several Mannopyranoside (MDMP) derivatives through direct regioselective acylation of methyl α-D-mannopyranoside (MDMP) derivatives. The synthesized compounds were characterized using various spectroscopic techniques, including FTIR, 1H-NMR, 13C-NMR, and elemental analysis. The antimicrobial activity of the compounds was evaluated against human and plant pathogenic bacteria and fungi, with compound 3 showing the most significant inhibitory effects on both gram-positive and gram-negative bacteria. The cytotoxicity of the compounds was assessed using the brine shrimp lethality test, with compound 2 exhibiting the lowest toxicity. Structure-activity relationship (SAR) analysis revealed that lauroyl and myristoyl acyl chains combined with mannopyranose were particularly effective against bacteria. Molecular docking studies were conducted to understand the binding interactions between the compounds and the active sites of Escherichia coli (4X08) and Aspergillus flavus (1R51), with compound 2 showing the strongest binding affinity for both proteins. Molecular dynamics simulations confirmed the stability of the compounds at the active sites of the proteins. ADMET predictions indicated that the compounds had good absorption, distribution, metabolism, excretion, and toxicity (ADMET) profiles. Overall, the results suggest that the MDMP derivatives have potential as antimicrobial agents and warrant further investigation for their therapeutic applications.This study investigates the synthesis, antimicrobial activity, molecular docking, molecular dynamics simulation, and ADMET properties of mannopyranoside derivatives as antimicrobial agents. The research aimed to explore the potential of these compounds against various pathogens, including bacteria and fungi. The study involved the synthesis of several Mannopyranoside (MDMP) derivatives through direct regioselective acylation of methyl α-D-mannopyranoside (MDMP) derivatives. The synthesized compounds were characterized using various spectroscopic techniques, including FTIR, 1H-NMR, 13C-NMR, and elemental analysis. The antimicrobial activity of the compounds was evaluated against human and plant pathogenic bacteria and fungi, with compound 3 showing the most significant inhibitory effects on both gram-positive and gram-negative bacteria. The cytotoxicity of the compounds was assessed using the brine shrimp lethality test, with compound 2 exhibiting the lowest toxicity. Structure-activity relationship (SAR) analysis revealed that lauroyl and myristoyl acyl chains combined with mannopyranose were particularly effective against bacteria. Molecular docking studies were conducted to understand the binding interactions between the compounds and the active sites of Escherichia coli (4X08) and Aspergillus flavus (1R51), with compound 2 showing the strongest binding affinity for both proteins. Molecular dynamics simulations confirmed the stability of the compounds at the active sites of the proteins. ADMET predictions indicated that the compounds had good absorption, distribution, metabolism, excretion, and toxicity (ADMET) profiles. Overall, the results suggest that the MDMP derivatives have potential as antimicrobial agents and warrant further investigation for their therapeutic applications.