ISOQUINOLINESULFONAMIDES, NOVEL AND POTENT INHIBITORS OF CYCLIC-NUCLEOTIDE DEPENDENT PROTEIN-KINASE AND PROTEIN KINASE-C
ISOQUINOLINESULFONAMIDES, NOVEL AND POTENT INHIBITORS OF CYCLIC-NUCLEOTIDE DEPENDENT PROTEIN-KINASE AND PROTEIN KINASE-C
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DOI：
10.1021/bi00316a032
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发表时间：
1984-01-01
期刊：
影响因子：
2.9
通讯作者：
SASAKI, Y
中科院分区：
文献类型：
--
作者：
HIDAKA, H;INAGAKI, M;SASAKI, Y
萘磺酰胺如 N-(6-氨基己基)-5-氯-1-萘磺酰胺 (W-7) 是有效的钙调蛋白 (CaM) 拮抗剂，在较高浓度下可作用于多种蛋白激酶。当萘环被异喹啉取代后，衍生物不再是CaM拮抗剂，而是保留了抑制蛋白激酶的能力，并且部分衍生物对某种蛋白激酶表现出选择性抑制作用。添加 10-6 M N-[2-(甲基氨基)乙基]-5-异喹啉磺酰胺 (H-8) 和 1-(5-异喹啉基磺酰基)-2-甲基哌嗪 (H-7) 可显着抑制 cAMP 依赖性、cGMP 依赖性和 Ca2+-磷脂依赖性（蛋白激酶 C）蛋白激酶。 H-8 是该系列抑制剂中活性最强的，与其他激酶相比，对环核苷酸依赖性蛋白激酶的抑制作用更显着，而具有磺酰哌嗪残基的衍生物 (H-7) 对蛋白激酶 C 的抑制作用最强。对于 cGMP 依赖性和 cAMP 依赖性蛋白激酶，H-8 的表观 Ki 值分别为 0.48 和 1.2 μM，而 H-7 对于蛋白激酶 C 的表观 Ki 值为6μM。全酶和无需酶激活剂就有活性的催化亚基（或片段）都对这些化合物敏感，具有相似的浓度依赖性，从而表明抑制作用归因于化合物与酶活性中心的直接相互作用，而不是与酶激活剂的直接相互作用。该抑制是可自由可逆的，并且是针对 ATP 的竞争型抑制和针对磷酸受体的非竞争型抑制。对 cAMP 依赖性蛋白激酶催化亚基的动力学研究表明，结构上与 ATP 无关的异喹啉磺酰胺与 ATP 竞争游离酶，但不与磷酸受体（即酶-ATP 复合物）相同的酶形式相互作用。这些衍生物的抑制效力似乎取决于异喹啉磺酰胺分子中末端氮所贡献的正电荷的位置和强度。本报告涉及异喹啉磺酰胺衍生物的制备和抑制机制，作为阐明蛋白激酶的体外和体内功能的可能工具。
Naphthalenesulfonamides such as N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7) are potent calmodulin (CaM) antagonists and act upon several protein kinases at higher concentration. When the naphthalene ring was replaced by isoquinoline, the derivatives were no longer CaM antagonists but retained the ability to inhibit protein kinases, and some of the derivatives exhibited selective inhibition toward a certain protein kinase. cAMP-dependent, cGMP-dependent and Ca2+-phospholipid-dependent (protein kinase C) protein kinases were inhibited significantly by addition of 10-6 M N-[2-(methylamino)ethyl]-5-isoquinolinesulfonamide (H-8) and 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7). H-8 was the most active of the inhibitors in this series and inhibited more markedly cyclic nucleotide dependent protein kinases, than other kinases, while the derivative with the sulfonylpiperazine residue (H-7) was the most potent in inhibiting protein kinase C. Apparent Ki values of H-8 were 0.48 and 1.2 .mu.M for cGMP-dependent and cAMP-dependent protein kinases, respectively, and the Ki value of H-7 for protein kinase C was 6 .mu.M. Both the holoenzyme and the catalytic subunit (or fragment), which is active without an enzyme activator, are susceptible to these compounds with a similar concentration dependency, thereby indicating that the inhibitory effect is attributed to the direct interaction of the compound with the active center of the enzyme but not with the enzyme activator. The inhibitions were freely reversible and of the competitive type with respect to ATP and of the noncompetitive type with respect to the phosphate acceptor. Kinetic studies with the catalytic subunit of cAMP-dependent protein kinase indicated that isoquinolinesulfonamides, structurally unrelated to ATP, compete with ATP for free enzyme but do not interact with the same enzyme form as does the phosphate acceptor (i.e., enzyme-ATP complex). The inhibitory potency of these derivatives seems to be dependent on the position and the strength of the plus charge contributed by the terminal nitrogen in the isoquinolinesulfonamide molecule. This report is concerned with the preparation and inhibitory mechanisms of isoquinolinesulfonamide derivatives, as possible tools for clarifying the in vitro and in vivo functions of protein kinases.