Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital, 32 Fruit Street, Boston, MA 02114, USA.
J Med Chem. 2012 Jul 26;55(14):6554-65. doi: 10.1021/jm300631e. Epub 2012 Jul 17.
We synthesized 5-allyl-1-methyl-5-(m-trifluoromethyl-diazirynylphenyl)barbituric acid (14), a trifluoromethyldiazirine-containing derivative of general anesthetic mephobarbital, separated the racemic mixture into enantiomers by chiral chromatography, and determined the configuration of the (+)-enantiomer as S by X-ray crystallography. Additionally, we obtained the (3)H-labeled ligand with high specific radioactivity. R-(-)-14 is an order of magnitude more potent than the most potent clinically used barbiturate, thiopental, and its general anesthetic EC(50) approaches those for propofol and etomidate, whereas S-(+)-14 is 10-fold less potent. Furthermore, at concentrations close to its anesthetic potency, R-(-)-14 both potentiated GABA-induced currents and increased the affinity for the agonist muscimol in human α1β2/3γ2L GABA(A) receptors. Finally, R-(-)-14 was found to be an exceptionally efficient photolabeling reagent, incorporating into both α1 and β3 subunits of human α1β3 GABA(A) receptors. These results indicate R-(-)-14 is a functional general anesthetic that is well-suited for identifying barbiturate binding sites on Cys-loop receptors.
我们合成了 5-烯丙基-1-甲基-5-(m-三氟甲基重氮基苯基)巴比妥酸(14),这是一种含三氟甲基重氮的一般麻醉药美芬妥因的衍生物,通过手性色谱将外消旋混合物分离成对映异构体,并通过 X 射线晶体学确定(+)-对映异构体的构型为 S。此外,我们还获得了具有高比放射性的(3)H 标记配体。R-(-)-14 的效力比最有效临床使用的巴比妥类药物硫喷妥钠强一个数量级,其全身麻醉 EC(50)接近丙泊酚和依托咪酯,而 S-(+)-14 的效力则低 10 倍。此外,在接近其麻醉效力的浓度下,R-(-)-14 既增强了 GABA 诱导的电流,又增加了人α1β2/3γ2L GABA(A)受体对激动剂 muscimol 的亲和力。最后,发现 R-(-)-14 是一种非常有效的光标记试剂,可掺入人α1β3 GABA(A)受体的α1 和β3 亚基中。这些结果表明,R-(-)-14 是一种功能性的全身麻醉药,非常适合鉴定 Cys 环受体上的巴比妥类药物结合位点。