Dibenzodiazepinone-type muscarinic receptor antagonists conjugated to basic peptides: Impact of the linker moiety and unnatural amino acids on M2R selectivity

Abstract

The family of human muscarinic acetylcholine receptors (MRs) is characterized by a high sequence homology among the five subtypes (M₁R-M₅R), being the reason for a lack of subtype selective MR ligands. In continuation of our work on dualsteric dibenzodiazepinone-type M₂R antagonists, a series of M₂R ligands containing a dibenzodiazepinone pharmacophore linked to small basic peptides was synthesized (64 compounds). The linker moiety was varied with respect to length, number of basic nitrogens (0-2) and flexibility. Besides proteinogenic basic amino acids (Lys, Arg), shorter homologues of Lys and Arg, containing three and two methylene groups, respectively, as well as D-configured amino acids were incorporated. The type of linker had a marked impact on M₂R affinity and also effected M₂R selectivity. In contrast, the structure of the basic peptide rather determined M₂R selectivity than M₂R affinity. For example, the most M₂R selective compound (UR-CG188, 89) with picomolar M₂R affinity (pK_i 9.60), exhibited a higher M₂R selectivity (ratio of K_i M₁R/M₂R/M₃R/M₄R/M₅R: 110:1:5200:55:2300) compared to the vast majority of reported M₂R preferring MR ligands. For selected ligands, M₂R antagonism was confirmed in a M₂R miniG protein recruitment assay.

Keywords: Basic amino acid; Dibenzodiazepinone derivative; Induced-fit docking; M2R antagonist; M2R selectivity; MR subtype selectivity; Muscarinic acetylcholine receptors; Peptide.