In silico identification and experimental validation of (-)-muqubilin A, a marine norterpene peroxide, as PPARalpha/gamma-RXRalpha agonist and RARalpha positive allosteric modulator

The nuclear receptors (NRs) RARalpha, RXRalpha, PPARalpha, and PPARgamma represent promising pharmacological targets for the treatment of neurodegenerative diseases. In the search for molecules able to simultaneously target all the above-mentioned NRs, we screened an in-house developed molecular database using a ligand-based approach, identifying (-)-Muqubilin (Muq), a cyclic peroxide norterpene from a marine sponge, as a potential hit. The ability of this compound to stably and effectively bind these NRs was assessed by molecular docking and molecular dynamics simulations. Muq recapitulated all the main interactions of a canonical full agonist for RXRalpha and both PPARalpha and PPARgamma, whereas the binding mode toward RAR? showed peculiar features potentially impairing its activity as a full agonist. Luciferase assays confirmed that Muq acts as a full agonist for RXRalpha, PPARalpha, and PPARgamma with an activity in the low- to sub-micromolar range. On the other hand, in the case of RAR, a very weak agonist activity was observed in the micromolar range. Quite surprisingly, we found that Muq is a positive allosteric modulator for RARalpha, as both luciferase assays and in vivo analysis using a zebrafish transgenic retinoic acid (RA) reporter line showed that co-administration of Muq with RA produced a potent synergistic enhancement of RARalpha activation and RA signaling.