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目的:观察7种黄酮类化合物对重组人蛋白激酶CK2全酶活性的影响并进行结构效应关系的分析。方法:将利用基因工程技术获得的重组人CK2α′及β亚基在体外等摩尔混合构成CK2全酶。通过测定药物作用后转移到CK2底物上的[γ-32P]ATP的32P的放射性活度,探讨黄酮类化合物对重组人CK2全酶活性的抑制作用,并采用半数效量概率单位法计算其IC50值。结果:杨梅黄酮、槲皮素、桑色素、毛地黄黄酮、莰非醇、芹菜苷配基和白杨黄素能明显抑制重组人CK2全酶活性,其IC50值分别是1.18,0.51,16.16,0.86,1.88,1.72和13.63μmol/L;其中杨梅黄酮、槲皮素、毛地黄黄酮、莰非醇和芹菜苷配基的作用效果均强于目前已知的CK2抑制剂DRB和A3;而桑色素和白杨黄素的作用效果接近于DRB。结构效应关系分析表明,C6,C3′和C4′上的-OH可能是黄酮类CK2抑制剂发挥抑制作用的药效基团;而在C3上去除1个-OH对其抑制效果基本没有影响;此外,C2′和C5′上的-OH可减弱黄酮类化合物对CK2的抑制作用。结论:黄酮类化合物对体外蛋白激酶CK2的抑制效果可能取决于其羟基的位置。
Objective: To observe the effect of seven flavonoids on the activity of recombinant human protein kinase CK2 holoenzyme and to analyze the relationship between the structural effects. Methods: Recombinant human CK2α ’and β subunits obtained by genetic engineering were equilibrated in vitro to form CK2 holoenzyme. The inhibitory effect of flavonoids on the activity of recombinant human CK2 holoenzyme was investigated by measuring the radioactivity of 32P of [γ-32P] ATP transferred to the substrate of CK2 after drug action. The half effective dose unit method was used to calculate its activity IC50 value. Results: Myricetin, quercetin, morin, lutein, non-alcohol, apigenin and chrysin could significantly inhibit the activity of recombinant human CK2 with IC50 values of 1.18, 0.51, 16.16, 0.86, 1.88 , 1.72 and 13.63μmol / L, respectively. The effect of myricetin, quercetin, foxglen, nonyl alcohol and apigenin were stronger than the currently known CK2 inhibitors DRB and A3; The effect of prime is close to DRB. The structural effect relationship analysis indicated that -OH on C6, C3 ’and C4’ may be the pharmacophore which could inhibit the action of flavone CK2 inhibitor. However, the removal of 1 -OH on C3 had no effect on the inhibitory effect. In addition, -OH on C2 ’and C5’ attenuated the inhibitory effect of flavonoids on CK2. Conclusion: The inhibitory effect of flavonoids on protein kinase CK2 in vitro may depend on the position of hydroxyl group.