9种蒽醌类衍生物对重组人蛋白激酶CK2活性的影响及构效浅析

目的观察9种蒽醌类衍生物对重组人蛋白激酶CK2全酶活性的影响,并进行构效分析。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,探讨蒽醌类衍生物对重组人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。结果米托蒽醌、2-羧基蒽醌、1,2-二氨蒽醌、1,8-二羟蒽醌和醌茜能明显抑制重组人CK2全酶活性,其IC50值分别是0.66、0.81、8.81、28.76和61.26μmol·L-1;其中米托蒽醌、2-羧基蒽醌和1,2-二氨蒽醌的作用效果均强于目前已知的CK2抑制剂DRB和A3。1-氨基蒽醌和1-氯蒽醌对CK2的作用效果较弱,其IC50值分别为143.38和221.18μmol·L-1;而蒽醌和2-乙基蒽醌对CK2的活性则没有明显的影响。构效分析表明:C2上的-COOH、C8位上的-OH以及C2上的-NH2可能分别是2-羧基蒽醌、1,8-二羟蒽醌和1,2-二氨基蒽醌的药效基团;而C2上的-CH2CH3和C1上的-Cl可能分别是2-乙基蒽醌和1-氯蒽醌的非必需基团。结论米托蒽醌、2-羧基蒽醌和1,2-二氨蒽醌是3种较强的重组人蛋白激酶CK2的抑制剂。

Effect and structure-activity study of nine kinds of anthraquinone derivatives on recombinant human protein kinase CK2 holoenzyme

Aim To observed the effects of nine kinds of anthraquinone derivatives on recombinant human protein kinase CK2 holoenzyme activity and investigated their structure-activity relationships. Methods Recombinant human protein kinase CK2α′ and β subunits were mixed at equal molar ratio to reconstitute CK2 holoenzyme. The CK2 activity was assayed by detecting incorporation of 32P of [γ-32P] ATP into the substrate for the inhibitory effect by anthraquinone derivatives. Calculation of IC50 values was performed according to the law of semi-effect-probit. Results Mitoxantrone, anthraquinone-2-carboxylic acid,1,2-diaminoanthraquinone,1,8-dihydroxyanthraquinone and quinizarin were shown to inhibit obviously recombinant CK2 holoenzyme activity in a concentration-dependent manner with IC50 values of 0.66、0.81、8.81、28.76 and 61.26 μmol·L-1, respectively; among that mitoxantrone, anthraquinone-2-carboxylic acid and 1,2-diaminoanthraquinone were more effective than DRB and A3, which were known as CK2 inhibitors in vitro. However, 1-aminoanthraquinone and 1-chloroanthraquinone undergo a drop in inhibitory efficiency, with IC50 values of 143.38, 221.18 μmol·L-1, respectively. While anthraquinone and 2-ethylanthraquinone did not change the CK2 activity fundamentally. Structure-activity study indicated that the major structural requirements for the potent inhibition of CK2 by anthraquinone-2-carboxylic acid,1,8-dihydroxyanthraquinone and 1,2-diaminoanthraquinone were carboxy group at position 2, hydroxyl group at position 8 and amino group at position 2, respectively. Differ from these requirements, as 2-ethylanthraquinone absence of a ethyl group at position 2 and 1-chloroanthraquinone absence of a chlorine group at position 1 did not modify their inhibitory potency. Conclusion Mitoxantrone, anthraquinone-2-carboxylic acid and 1,2-diaminoanthraquinone were three stronger inhibitors of recombinant human protein kinase CK2 in vitro.