Repurposing eugenol and cinnamaldehyde as potent antimicrobial agents: A comprehensive in-vitro and in-silico study.

Multi-drug-resistant (MDR) pathogens represent a critical global health threat, necessitating the development of novel antimicrobial agents with broad-spectrum activity and minimal toxicity. This study investigates the antimicrobial and anti-biofilm properties of 4-Allyl-2-methoxyphenol (eugenol, EU) and (E)-3-Phenylprop-2-enal (cinnamaldehyde, CN) against 19 clinically significant pathogens through a combination of in-vitro assays and in-silico analyses. EU displayed remarkable activity, particularly against Aspergillus niger (20.5 ± 0.5 mm), and strong binding affinities with key protein targets, including peptide deformylase and β-carbonic anhydrase, with binding free energies (ΔG) ranging from -12.75 to -0.60 kcal/mol. CN exhibited exceptional activity against Staphylococcus epidermidis (29.6 ± 0.4 mm) and Candida albicans (36.6 ± 0.4 mm), supported by a significant binding affinity with β-carbonic anhydrase (ΔG: -5.23 kcal/mol). Dissociation constants (Kd) derived from MM-GBSA analyses indicated EU's strong inhibitory potential with nano- to picomolar Kd values, directly correlating with low IC50 values. CN demonstrated moderate inhibitory activity with Kd in the micromolar range. Molecular dynamics (MD) simulations confirmed the stability of these protein-ligand complexes, revealing critical hydrophobic interactions, such as those involving PHE122, that contributed to binding stabilization. ADMET profiling further underscored the favorable pharmacokinetics and safety of both compounds. These findings establish EU and CN as promising candidates for antimicrobial therapy, with potential applications in combating MDR pathogens and biofilm-associated infections. The complementary strengths of EU and CN warrant further structural optimization and combination studies, offering new avenues in the development of next-generation antimicrobial agents.