Singh L, Donald A E, Foster A C, Hutson P H, Iversen L L, Iversen S D, Kemp J A, Leeson P D, Marshall G R, Oles R J
Merck Sharp & Dohme Research Laboratories, Neuroscience Research Centre, Harlow, Essex, United Kingdom.
Proc Natl Acad Sci U S A. 1990 Jan;87(1):347-51. doi: 10.1073/pnas.87.1.347.
The antagonist effect of (+/-)-3-amino-1-hydroxypyrrolid-2-one (HA-966) at the N-methyl-D-aspartate (NMDA) receptor occurs through a selective interaction with the glycine modulatory site within the receptor complex. When the enantiomers of (+/-)-HA-966 were resolved, the (R)-(+)-enantiomer was found to be a selective glycine/NMDA receptor antagonist, a property that accounts for its anticonvulsant activity in vivo. In contrast, the (S)-(-)-enantiomer was only weakly active as an NMDA-receptor antagonist, but nevertheless it possessed a marked sedative and muscle relaxant action in vivo. In radioligand binding experiments, (+)-HA-966 inhibited strychnine-insensitive [3H]glycine binding to rat cerebral cortex synaptic membranes with an IC50 of 12.5 microM, whereas (-)-HA-966 had an IC50 value of 339 microM. In electrophysiological experiments, (+)-HA-966 selectively antagonized NMDA receptor responses in rat cortical slices, whereas the (-)-enantiomer was much weaker. On cultured cortical neurones (+)-HA-966 inhibited glycine-potentiated NMDA responses with an IC50 = 13 microM compared with (-)-HA-966, which has an IC50 = 708 microM. In agreement with findings with racemic HA-966, even high concentrations of (+)-HA-966 did not completely inhibit NMDA responses, suggesting that (+)-HA-966 is a low-efficacy partial agonist. (+)-HA-966 produced parallel shifts to the right of the glycine concentration curve for potentiation of NMDA responses, resulting in an estimated pKb = 5.6. In mice, (+)-HA-966 antagonized sound and N-methyl-DL-aspartic acid (NMDLA)-induced seizures with ED50 values of 52.6 mg/kg of body weight (i.p.) and 900 mg/kg (i.v.), respectively. The coadministration of D-serine dose-dependently (10-100 micrograms into the cerebral ventricles per mouse) antagonized the anticonvulsant effect of a submaximal dose of (+)-HA-966 (100 micrograms administered directly into the cerebral ventricles) against NMDLA-induced seizures. The sedative/ataxic effect of racemic HA-966 was mainly attributable to the (-)-enantiomer, which was greater than 25-fold more potent than the (+)-enantiomer. It is suggested that, as in the case of the sedative gamma-butyrolactone, disruption of striatal dopaminergic mechanisms may be responsible for this action.
(±)-3-氨基-1-羟基吡咯烷-2-酮(HA-966)对N-甲基-D-天冬氨酸(NMDA)受体的拮抗作用是通过与受体复合物内的甘氨酸调节位点选择性相互作用而发生的。当拆分(±)-HA-966的对映体时,发现(R)-(+)-对映体是一种选择性甘氨酸/NMDA受体拮抗剂,这一特性解释了其在体内的抗惊厥活性。相比之下,(S)-(-)-对映体作为NMDA受体拮抗剂的活性较弱,但在体内仍具有显著的镇静和肌肉松弛作用。在放射性配体结合实验中,(+)-HA-966抑制士的宁不敏感的[³H]甘氨酸与大鼠大脑皮质突触膜的结合,IC50为12.5微摩尔,而(-)-HA-966的IC50值为339微摩尔。在电生理实验中,(+)-HA-966选择性拮抗大鼠皮质切片中的NMDA受体反应,而(-)-对映体的作用则弱得多。在培养的皮质神经元上,(+)-HA-966抑制甘氨酸增强的NMDA反应,IC50 = 13微摩尔,而(-)-HA-966的IC50 = 708微摩尔。与消旋HA-966的研究结果一致,即使高浓度的(+)-HA-966也不能完全抑制NMDA反应,表明(+)-HA-966是一种低效部分激动剂。(+)-HA-966使甘氨酸浓度曲线平行右移以增强NMDA反应,估计pKb = 5.6。在小鼠中,(+)-HA-966拮抗声音和N-甲基-DL-天冬氨酸(NMDLA)诱导的惊厥,ED50值分别为52.6毫克/千克体重(腹腔注射)和900毫克/千克(静脉注射)。共同给予D-丝氨酸剂量依赖性地(每只小鼠向脑室注射10 - 100微克)拮抗亚最大剂量的(+)-HA-966(直接向脑室注射100微克)对NMDLA诱导惊厥的抗惊厥作用。消旋HA-966的镇静/共济失调作用主要归因于(-)-对映体,其效力比(+)-对映体大25倍以上。有人认为,与镇静性γ-丁内酯的情况一样,纹状体多巴胺能机制的破坏可能是这种作用的原因。