Novel Bivalent 5-HT_2A Receptor Antagonists Exhibit High Affinity and Potency in Vitro and Efficacy in Vivo

The 5-HT_2A receptor (5-HT_2AR) plays an important role in various neuropsychiatric disorders, including substance use disorder and schizophrenia. Homodimerization of this receptor has been suggested, but tools that allow direct assessment of the relevance of the 5-HT_2AR:5-HT_2AR homodimer in these disorders are necessary. We chemically modified the selective 5-HT_2AR antagonist M100907 to synthesize a series of homobivalent ligands connected by ethylene glycol linkers of varying lengths that may be useful tools for probing 5-HT_2AR:5-HT_2AR homodimer function. We tested these molecules for 5-HT_2AR antagonist activity in a cell line stably expressing the functional 5-HT_2AR and quantified a downstream signaling target, activation (phosphorylation) of extracellular regulated kinases 1/2 (ERK_1/2), in comparison to in vivo efficacy of altering spontaneous or cocaine-evoked locomotor activity in rats. All of the synthetic compounds inhibited 5-HT-mediated phosphorylation of ERK_1/2 in the cellular signaling assay; the potency of the bivalent ligands varied as a function of linker length, with the intermediate linker lengths being the most potent. The K_i values for the binding of bivalent ligands to 5-HT_2AR were only slightly lower than the values for the parent (+)-M100907 compound, but significant selectivity for 5-HT_2AR over 5-HT_2BR or 5-HT_2CR binding was retained. In addition, the 11-atom-linked bivalent 5-HT_2AR antagonist (2 mg/kg, intraperitoneally) demonstrated efficacy on par with that of (+)-M100907 in inhibiting cocaine-evoked hyperactivity. As we develop further strategies for ligand-evoked receptor assembly and analyses of diverse signaling and functional roles, these novel homobivalent 5-HT_2AR antagonist ligands will serve as useful in vitro and in vivo probes of 5-HT_2AR structure and function.