Structure-guided discovery of phenyl-carbamates as novel dual TRPV1 antagonists/FAAH inhibitors.

The pathophysiology of pain involves multiple signaling pathways, limiting the efficacy and safety of single-target analgesics. Dual-target agents that simultaneously inhibit fatty acid amide hydrolase (FAAH) and antagonize transient receptor potential vanilloid 1 (TRPV1) offer a promising strategy to enhance analgesic efficacy while minimizing side effects. Through structural hybridization of pharmacophoric elements from FAAH inhibitors with either classical or newly discovered TRPV1 antagonists, we designed and synthesized multiple series of dual-active ligands. Among these, compound 31 emerged as the lead candidate, demonstrating potent dual activity with an IC50 of 0.57 μM against FAAH and 18.81 nM against TRPV1 in vitro. In vivo, compound 31 demonstrated significant analgesic effects in the formalin test and pronounced anti-inflammatory activity in a carrageenan-induced rat paw edema model. Importantly, it did not induce TRPV1-mediated hyperthermia at therapeutic dose. Target engagement studies confirmed in vivo modulation of both FAAH and TRPV1, and molecular docking and molecular dynamics study revealed stable binding conformations with each target. These findings establish compound 31 as a novel dual-target scaffold with therapeutic potential for pain management.