Background: Cuachalalate (Amphipterygium adstringens) is one of Mexico’s most commercialised medicinal plants, and its natural populations are mainly found in this country. The high demand for stem bark (SB) has risked its survival, making it necessary to develop sustainable strategies.Aim: This study aimed to identify bioactive compounds in branch bark (BB) and evaluate their antipathogenic and antivirulence capacities.Setting: The study was conducted in the low deciduous forest (LDF) of the State of Puebla, Mexico.Methods: Bioactive metabolites in BB were profiled by high-performance thin-layer chromatography (HPTLC). The chemical composition of aqueous bark extracts was scrutinised by high-performance liquid chromatography-mass spectrometry (HPLC-EM-SQ-TOF system) and molecular network analysis. The folk method’s antipathogenic capacity was determined in a burn model in mice infected with Pseudomonas aeruginosa. Finally, the antivirulence properties of bark organic extracts were determined.Results: The BB contained masticadienoic, 3α-hydroxymasticadienoic and anacardic acids, also present in SB. The aqueous extracts showed the presence of flavonoids, catechins and saponins in both barks. The folk method using BB extracts significantly reduced mouse mortality and prevented sepsis development. This might be related to the capability of extracts to block the production of bacteria virulence factors.Conclusion: The similarity in bioactive metabolites and biological activity between SB and BB of cuachalalate suggests that using BB as a medicinal agent could be a practical and sustainable strategy. This approach could potentially prevent the overexploitation of cuachalalate’s SB, contributing to its conservation.Contribution: This study proposes a sustainable strategy for using cuachalalate as a medicinal agent using BB, a renewable source.