In this context, the aim of this work was to predict the bioactive conformation of endogenous oleoyl-substituted LysoPS(18:1) (1, Figure ) toward the receptor GPR34 by comparing the bioactivities and accessible conformational spaces of four LysoPS analogues (Figure ) conformationally restricted at the glycerol moiety. Previously, we have predicted the selectivity of inhibitor complexes with Bcl-2 and Bcl-XL and the mechanism the resistance of influenza cap-dependent endonuclease against Baloxavir marboxil using molecular dynamics (MD) simulations. Comparing to these simulations, the sampling of accessible conformational space is more difficult because it requires to sample entire conformational space of the target ligand. Hence, this study carried out long-duration replica exchange molecular dynamics (REMD) as well as metadynamics (MTD) simulations to sample comprehensive conformational space of LysoPS and its analogues. The analogues studied in this work were selected from those synthesized during our previous experimental structure-activity relationship study, which established that agonistic potency toward GPR34 was dependent on the regio-and stereochemistry of the glycerol moiety (Figure and Table ). We focused here on the effect of key dihedral angles of the glycerol moiety on the agonistic activity (Figure ).