Thermodynamics of the interactions of tricyclic drugs with binding sites for [3H]imipramine in mouse cerebral cortex.
Thermodynamics of the interactions of tricyclic drugs with binding sites for [3H]imipramine in mouse cerebral cortex.
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三环类药物与小鼠大脑皮质中[3H]丙咪嗪结合位点相互作用的热力学。
DOI：
10.1016/0006-2952(84)90030-3
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发表时间：
1984
影响因子：
5.8
通讯作者：
Lajtha,A
中科院分区：
文献类型：
--
作者：
Reith,ME;Sershen,H;Lajtha,A
材料和方法以下药物是所示公司的捐赠品：阿米替林，Merck Sharp&Dohme;盐酸氯丙咪嗪，Geigy Pharmaceuticals;盐酸可卡因，Mallinckrodt;盐酸地昔帕明，USV Laboratory;氟西汀，Lilly Research Laboratories;盐酸丙咪嗪，Geigy Pharmaceuticals;和去甲丙美定，Astra。盐酸5-羟色胺购自Sigma，IV-[甲基-3H]丙咪嗪盐酸盐（75 Ci/mmole）购自新英格兰核能公司。制备来自成年雄性BALB/cBy小鼠（20-25 g，杰克逊实验室，Bar Harbor，ME）大脑皮层的膜，并通过Langer等人的方法立即测定[3 H]丙咪嗪的结合。[5]如前所述[13]。孵育混合物含有50 mM Tris-HCl、100 mM NaCl和5 mM KCl，在25 ° C下缓冲至pH 7.4。通过用单歧管MiIIipore过滤装置快速过滤终止结合测定。~ 3 H]丙咪嗪的非特异性结合（见下文）占总结合的2@-32%。化合物抑制[3 H]丙咪嗪结合的ic 50 * 值由在三个单独的组织制备物中一式三份测定的四种或五种浓度的化合物获得的抑制的对数概率单位分析计算。用Cheng和Prusoff校正[14]从nk值计算nk值：Kr= ICKY，/（1+[L] jKl，）。[L]游离（3 H）丙咪嗪的浓度在3”处为1.7 nM，在10”和14”处为2.2 nM，在17”和25”处为3.0 nM。通过Scatchard分析测定的[3 H]丙咪嗪的Ki值在3”处为21 nM，在10”处为41 nM。在17”处91 nM，在25”处145 nM。由抑制实验计算的丙咪嗪的KD值（见表1）是相似的。结合的标准吉布斯自由能变化（AG）从AG”=
Materials and methodsThe following drugs were the donations of the companies indicated: amitriptyline, Merck Sharp & Dohme; chlorimipramine hydrochloride, Geigy Pharmaceuticals; cocaine hydrochloride, Mallinckrodt; desipramine hydrochloride, USV Laboratory; fluoxetine, Lilly Research Laboratories; imipramine hydrochloride, Geigy Pharmaceuticals; and norzimelidine, Astra. Serotonin hydrochloride was purchased from Sigma and IV-[methyl-3H] imipramine hydrochloride (75 Ci/mmole) from New England Nuclear. Membranes from cerebral cortex of adult male BALB/cBy mice (20-25 g, Jackson Laboratories, Bar Harbor, ME) were prepared and assayed immediately for binding of [3H] imipramine by the method of Langer et al.[5] as described previously [13]. The incubation mixtures contained 50 mM Tris-HCI, 100 mM NaCl, and 5 mM KCl, buffered at pH 7.4 at 25”. Binding assays were terminated by rapid filtration with a single-manifold Miilipore filtration apparatus. Nonspecific binding of~ 3H] imipramine (see below) contributed 2@-32% of total binding. The icso* values of compounds in inhibiting the binding of [3H] imipramine were calculated from log-probit analysis of inhibition obtained with four or five concentrations of the compound assayed in triplicate in three separate tissue preparations. iu, values were calculated from the n& values with the Cheng and Prusoff correction [14]: Kr= ICKY,/(1+[L] jKl,).[L], the concentration of free (3H] imipramme, was 1.7 nM at 3”, 2.2 nM at lo” and 14”, and 3.0 nM at 17” and 25”. K, values for [3H] imipramine, determined by Scatchard analysis, were 21 nM at 3”, 41 nM at 10”. 91 nM at 17”, and 145 nM at 25”. KD values for imipramine calculated from inhibition experiments (see Table 1) were similar. The standard Gibbs free energy change (AG) of binding was calculated for 25’from AG”=