Enantiomers of cis-constrained and flexible 2-substituted GABA analogues exert opposite effects at recombinant GABAC receptors

The effects of the enantiomers of a number of flexible and cis-constrained GABA analogues were tested on GABAC receptors expressed in Xenopus laevis oocytes using two-electrode voltage-clamp electrophysiology. (1S,2R)-cis-2-Aminomethylcyclopropane-1-carboxylic acid ((+)-CAMP), a potent and full agonist at the ρ1 (EC50 ≈ 40 μM, Imax ≈ 100%) and ρ 2 (EC50 ≈ 17 μM, Imax ≈ 100%) receptor subtypes, was found to be a potent partial agonist at ρ3 (EC50 ≈ 28 μM, Imax ≈ 70%). (1R,2S)-cis-2-Aminomethylcyclopropane-1-carboxylic acid ((−)-CAMP), a weak antagonist at human ρ1 (IC50 ≈ 890 μM) and ρ2 (IC50 ≈ 400 μM) receptor subtypes, was also found to be a moderately potent antagonist at rat ρ3 (IC50 ≈ 180 μM). Similarly, (1R,4S)-4-aminocyclopent-2-ene-1-carboxylic acid ((+)-ACPECA) was a full agonist at ρ1 (EC50 ≈ 135 μM, Imax ≈ 100%) and ρ2 (EC50 ≈ 60 μM, Imax ≈ 100%), but only a partial agonist at ρ3 (EC50 ≈ 112 μM, Imax ≈ 37 %), while (1S,4R)-4-aminocyclopent-2-ene-1-carboxylic acid ((−)-ACPECA) was a weak antagonist at all three receptor subtypes (IC50 >> 300 μM). 4-Amino-(S)-2-methylbutanoic acid ((S)-2MeGABA) and 4-amino-(R)-2-methylbutanoic acid ((R)-2MeGABA) followed the same trend, with (S)-2MeGABA acting as a full agonist at the ρ1 (EC50 ≈ 65 μM, Imax ≈ 100%), and ρ2 (EC50 ≈ 20 μM, Imax ≈ 100%) receptor subtypes, and a partial agonist at ρ3 (EC50 ≈ 25 μM, Imax ≈ 90%). (R)-2MeGABA, however, was a moderately potent antagonist at all three receptor subtypes (IC50 ≈ 16 μM at ρ1, 125 μM at ρ2 and 35 μM at ρ3). On the basis of these expanded biological activity data and the solution-phase molecular structures obtained at the MP2/6-31+G* level of ab initio theory, a rationale is proposed for the genesis of this stereoselectivity effect.

The effects of the enantiomers of a number of flexible and cis-constrained GABA analogues are tested on GABAC receptors expressed in Xenopus Laevis oocytes using two-electrode voltage-clamp electrophysiology. Different enantiomers are found to have opposite biological activities, with the (S) and (+) enantiomers possessing antagonist activity and the (R) and (−) enantiomers possessing antagonist activity. A novel stereoselective binding mechanism is proposed to explain this effect.Figure optionsDownload as PowerPoint slide