对非循环细胞有活性的安吖啶氨基甲酸酯类似物：对拓扑异构酶IIα和β的相对活性

Carbamate analogues of amsacrine active against non-cycling cells: relative activity against topoisomerases IIalpha and beta.

作者信息

Turnbull R M, Meczes E L, Perenna Rogers M, Lock R B, Sullivan D M, Finlay G J, Baguley B C, Austin C A

机构信息

School of Biochemistry and Genetics, The Medical School, The University of Newcastle upon Tyne, Newcastle upon Tyne NE2 4HH, UK.

出版信息

Cancer Chemother Pharmacol. 1999;44(4):275-82. doi: 10.1007/s002800050978.

DOI:10.1007/s002800050978

PMID:10447574

Abstract

PURPOSE

Methyl N-(4'-(9-acridinylamino)-phenyl)carbamate hydrochloride (AMCA) and methyl N-(4'-(9-acridinylamino)-2-methoxyphenyl)carbamate hydrochloride (mAMCA) are analogues of the topoisomerase II (topo II) poison amsacrine, and are distinguished from amsacrine by their high cytotoxicity towards non-cycling cells. Since mammalian cells contain two forms (alpha and beta) of topo II and the alpha isoform is down-regulated in non-cycling cells, we have considered whether these carbamate analogues target topo IIbeta selectively.

METHODS

A drug permeable yeast strain (JN394 top2-4) was transformed using a shuttle vector containing either human top2alpha, human top2alpha or yeast top2 under the control of a GAL1 promoter. The strain was analysed at a non-permissive temperature, where only the plasmid-borne topo II was active.

RESULTS

AMCA and mAMCA produced comparable levels of cell killing with human DNA topo IIalpha, human DNA topo IIbeta and yeast DNA topo II. Two other acridine derivatives N-[2-(dimethylamino)ethyl]acridine-4-carboxamide (DACA) and its 7-chloro derivative, which like AMCA and mAMCA are able to overcome multidrug resistance mechanisms, were much more active against human DNA topo IIalpha than against human DNA topo IIbeta and yeast DNA topo II. A series of mutant Chinese hamster and human lines with defined topo lesions, including the HL60/MX2 line that lacks topo IIbeta expression, was also used to compare resistance to amsacrine, AMCA and etoposide. Loss of topo IIbeta activity had a greater effect on amsacrine and AMCA than on etoposide. Resistance of murine Lewis lung cultures in exponential and plateau phase was also measured. Loss of topo IIalpha activity, as measured in both mutant cells expressing lower amounts of enzyme and in cells in plateau phase, resulted in concomitant acquisition of resistance that was greatest for etoposide and least for AMCA.

CONCLUSION

We conclude that the carbamate analogues of amsacrine recognize both topo IIalpha and beta in cells.

摘要

目的

盐酸N-（4'-（9-吖啶基氨基）-苯基）氨基甲酸甲酯（AMCA）和盐酸N-（4'-（9-吖啶基氨基）-2-甲氧基苯基）氨基甲酸甲酯（mAMCA）是拓扑异构酶II（topo II）毒药安吖啶的类似物，它们与安吖啶的区别在于对非循环细胞具有高细胞毒性。由于哺乳动物细胞含有两种形式（α和β）的topo II，且α同工型在非循环细胞中下调，我们考虑了这些氨基甲酸酯类似物是否选择性靶向topo IIβ。

方法

使用在GAL1启动子控制下含有人类top2α、人类top2α或酵母top2的穿梭载体转化一种药物可渗透的酵母菌株（JN394 top2-4）。在非允许温度下分析该菌株，此时只有质粒携带的topo II具有活性。

结果

AMCA和mAMCA对人类DNA topo IIα、人类DNA topo IIβ和酵母DNA topo II产生的细胞杀伤水平相当。另外两种吖啶衍生物N-[2-（二甲基氨基）乙基]吖啶-4-甲酰胺（DACA）及其7-氯衍生物，与AMCA和mAMCA一样能够克服多药耐药机制，它们对人类DNA topo IIα的活性比对人类DNA topo IIβ和酵母DNA topo II的活性高得多。还使用了一系列具有明确拓扑异构酶损伤的突变中国仓鼠和人类细胞系，包括缺乏topo IIβ表达的HL60/MX2细胞系，来比较对安吖啶、AMCA和依托泊苷的耐药性。topo IIβ活性丧失对安吖啶和AMCA的影响比对依托泊苷的影响更大。还测量了处于指数期和平原期的小鼠Lewis肺癌培养物的耐药性。在表达较低量酶的突变细胞和平原期细胞中测量到的topo IIα活性丧失导致同时获得耐药性，对依托泊苷的耐药性最大，对AMCA的耐药性最小。