IRIC - Institute for Research in Immunology and Cancer

Histamine acts through four different receptors (H₁R-H₄R), the H₃R and H₄R being the most explored in the last years as drug targets. The H₃R is a potential target to treat narcolepsy, Parkinson’s disease, epilepsy, schizophrenia and several other CNS-related conditions, while H₄R blockade leads to anti-inflammatory and immunomodulatory effects. Our group has been exploring the dihydrobenzofuranyl-piperazines (LINS01 series) as human H₃R/H₄R ligands as potential drug candidates. In the present study, a set of 12 compounds were synthesized from adequate (dihydro)benzofuran synthons through simple reactions with corresponding piperazines, giving moderate to high yields. Four compounds (1b, 1f, 1g and 1h) showed high hH₃R affinity (pK_i > 7), compound 1h being the most potent (pK_i 8.4), and compound 1f showed the best efficiency (pKi 8.2, LE 0.53, LLE 5.85). BRET-based assays monitoring Gα_i activity indicated that the compounds are potent antagonists. Only one compound (2c, pK_i 7.1) presented high affinity for hH₄R. In contrast to what was observed for hH₃R, it showed partial agonist activity. Docking experiments indicated that bulky substituents occupy a hydrophobic pocket in hH₃R, while the N-allyl group forms favorable interactions with hydrophobic residues in the TM2, 3 and 7, increasing the selectivity towards hH₃R. Additionally, the importance of the indole NH in the interaction with Glu5.46 from hH₄R was confirmed by the modeling results, explaining the affinity and agonistic activity of compound 2c. The data reported in this work represent important findings for the rational design of future compounds for hH₃R and hH₄R.