The opportunistic pathogen Pseudomonas aeruginosa is one of the “critical priority patho-gens” due to its multidrug resistance to a wide range of antibiotics. Its ability to invade and damage host tissues is due to the use of quorum sensing (QS) to collectively produce a plethora of virulence factors. Inhibition of QS is an attractive strategy for new antimi-crobial agents because it disrupts the initial events of infection without killing the path-ogen. Highly diverse microorganisms as endophytes represent an under-explored source of bioactive natural products, offering opportunities for the discovery of novel QS inhib-itors (QSI). In the present work, the objective was to explore selective QSIs within a unique collection of fungal endophytes isolated from the tropical palm Astrocaryum sci-ophilum. The fungi were cultured, extracted, and screened for their antibacterial and spe-cific anti-QS activities against P. aeruginosa. The endophytic strain Lasiodiplodia venezue-lensis was prioritized for scaled-up fractionation for its selective activity, leading to the isolation of eight compounds in a single step. Among them, two pyran-derivatives were found to be responsible for the QSI activity, with an effect on some QS-regulated viru-lence factors. Additional non-targeted metabolomic studies on P. aeruginosa documented their effects on the production of various virulence-related metabolites.