Hering S, Hughes A D, Timin E N, Bolton T B
Department of Pharmacology and Clinical Pharmacology, St. George's Hospital Medical School, London, United Kingdom.
J Gen Physiol. 1993 Mar;101(3):393-410. doi: 10.1085/jgp.101.3.393.
The actions of the optical enantiomers of BAY K 8644 and Sandoz 202,791 were studied on barium inward currents recorded using the whole-cell configuration of the patch clamp technique from enzymatically isolated smooth muscle cells from the rabbit ear artery. The enantiomers were applied by bath perfusion or rapidly by a concentration jump technique, which enabled the study of drug action under equilibrium and nonequilibrium conditions. A larger effect of agonists was seen on peak inward current in 110 mM Ba when small rather than large depolarizations were applied. The midpoint voltage of the steady-state inactivation curve of IBa was -12.8 +/- 1.9 mV (n = 4) in the absence of drug, -16.4 +/- 2.5 mV (n = 4) in 1 microM (+)202,791, and -31.4 +/- 0.4 mV (n = 4) in 1 microM (-)202,791. The rate of onset of action of the agonist and antagonist enantiomers of BAY K 8644 and Sandoz 202,791 was studied by rapid application during 20-ms depolarizing steps from different holding potentials to +30 mV at 1 or 0.2 Hz. The drugs were applied as concentration jumps between two single pulses of a pulse train. The rates of onset of drug action on peak IBa during a 1-Hz pulse train were concentration dependent over the range of 100 nM-3 microM for both (+) and (-)202,791. The rate of onset of inhibition of peak current by antagonist enantiomers was not significantly influenced by the test pulse frequency. At a holding potential of -60 mV, the onset rate of the increase in peak IBa on application of 1 microM of agonist enantiomers (+)202,791 or (-)BAY K 8644 during a train of pulses occurred with mean time constants of 2.1 +/- 0.7 s (n = 7) and 2.3 +/- 0.2 s (n = 4), respectively. The onset of current increase on application of 1 microM (+)202,791 during a single voltage clamp step to 20 mV was faster, with a mean time constant of 380 +/- 80 ms (n = 3).
使用膜片钳技术的全细胞模式,从兔耳动脉酶解分离的平滑肌细胞记录钡内向电流,研究了BAY K 8644和山德士202,791光学对映体的作用。对映体通过浴槽灌注或采用浓度阶跃技术快速施加,这使得能够在平衡和非平衡条件下研究药物作用。当施加小的而非大的去极化时,激动剂对110 mM钡中的内向电流峰值有更大影响。在无药物时,IBa稳态失活曲线的中点电压为-12.8±1.9 mV(n = 4),在1 μM(+)202,791中为-16.4±2.5 mV(n = 4),在1 μM(-)202,791中为-31.4±0.4 mV(n = 4)。通过在1或0.2 Hz下从不同的钳制电位进行20 ms去极化步骤快速施加,研究了BAY K 8644和山德士202,791激动剂和拮抗剂对映体的作用起效速率。药物作为脉冲序列两个单脉冲之间的浓度阶跃施加。在1 Hz脉冲序列期间,(+)和(-)202,791对IBa峰值的药物作用起效速率在100 nM - 3 μM范围内均呈浓度依赖性。拮抗剂对映体对峰值电流抑制的起效速率不受测试脉冲频率的显著影响。在-60 mV的钳制电位下,在脉冲序列期间施加1 μM激动剂对映体(+)202,791或(-)BAY K 8644时,IBa峰值增加的起效速率的平均时间常数分别为2.1±±0.7 s(n = 7)和2.3±0.2 s(n = 4)。在单次电压钳制步骤至20 mV期间施加1 μM(+)202,791时电流增加的起效更快,平均时间常数为380±80 ms(n = 3)。