Antinociceptive Effects of a Novel α2/α3-Subtype Selective GABA_A Receptor Positive Allosteric Modulator

Pain remains a challenging clinical condition and spinal GABA_A receptors are crucial modulators of pain processing. α2/α3-subtype GABA_A receptors mediate the analgesic actions of benzodiazepines. Positive allosteric modulators (PAMs) at α2/α3-subtype GABA_A receptors may have analgesic potential. Here we report a new selective α2/α3-subtype GABA_A receptor PAM in in vitro and in vivo pain assays. KRM-II-81 demonstrated similar efficacy at α1/α2/α3 GABA_A receptors and negligible efficacy at α4/α5/α6 GABA_A receptors, with α2 and α3-subtypes being 17- and 28-fold more potent than α1 subtypes in HEK-293T cells expressing GABA_A receptors with different α subunits. In contrast, KRM-II-18B showed significant efficacy at α1/α2/α3/ α5 subtypes, with similar potency at α1/α2/α3 subtypes. Both PAMs and morphine dose-dependently decreased 0.6% acetic acid- and 0.32% lactic acid-induced writhing. The effects of both PAMs were reversed by the benzodiazepine receptor antagonist flumazenil, confirming their action at the benzodiazepine binding site of GABA_A receptors. Both PAMS and morphine all dose-dependently reversed 0.32% lactic acid (but not 0.6% acetic acid) induced suppression of nesting behavior. Acetaminophen, but not the PAMs, reversed acid-depressed locomotor activity. Combined, these findings suggest that KRM-II-81 is a selective α2/α3 subtype GABA_A PAM with significant antinociceptive effects in chemical stimulation-induced pain in mice.