Integrated In Vitro and In Silico Evaluation of the Antimicrobial and Cytotoxic Potential of Calotropis procera Leaf Ethanolic Extract: From GC-MS Profiling to Molecular Docking and Dynamics

TL;DR

This study explores the antimicrobial and safety properties of a plant extract from Calotropis procera, showing strong activity against bacteria and potential for drug development.

Contribution

The first report on Colombian C. procera demonstrating anti-Staphylococcus activity and MurG-targeted cardenolides.

Findings

EE-CP showed 93% inhibition of Staphylococcus aureus and 52% inhibition of Escherichia coli compared to amoxicillin.

Molecular docking and dynamics simulations confirmed strong binding of strophanthidin and NCGC00384918 to MurG.

EE-CP exhibited acceptable erythrocyte compatibility with <10% hemolysis at 1.5 mg mL−1.

Abstract

Calotropis procera, a drought-tolerant shrub widely used in folk medicine, was evaluated for its antimicrobial potential and safety using an integrative in vitro/in silico workflow. Ethanolic leaf extract (EE-CP) displayed a dose-dependent inhibition of Staphylococcus aureus ATCC 2913 and Escherichia coli ATCC 35218, reaching 93% and 52% of the amoxicillin control, respectively (MIC 207 µg mL−1 and 149 µg mL−1). GC-MS and LC-HRMS profiling revealed cardenolides (strophanthidin, gitoxigenin) and indole derivatives as major constituents. Pharmacophore mapping highlighted the essential glycosyltransferase MurG as a likely bacterial target; molecular docking showed that strophanthidin and NCGC00384918 bind MurG more strongly than the native substrate UDP-GlcNAc (ΔG ≤ −9.4 kcal mol−1), a result corroborated by 100 ns molecular dynamics simulations and MM-PBSA binding energies (−96.4 and −49.3 kcal mol−1). EE-CP caused <10% hemolysis up to 1.5 mg mL−1 and exhibited LC50 values of 302 µg mL−1 (human lymphocytes) and 247 µg mL−1 (BHK-21 cells), indicating a narrow but exploitable therapeutic window. Collectively, these findings constitute the first report on Colombian C. procera demonstrating potent anti-Staphylococcus activity, MurG-targeted cardenolides, and acceptable erythrocyte compatibility. This study supports EE-CP as a promising source of lead molecules and antibiotic adjuvants, warranting guided fractionation and in vivo validation to optimize efficacy and mitigate cytotoxicity.

Figures