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


CONCLUSIONS
We conclude that the carbamate analogues of amsacrine recognize both topo IIalpha and beta in cells.
Turnbull
R M
RM
School of Biochemistry and Genetics, The Medical School, The University of Newcastle upon Tyne, Newcastle upon Tyne NE2 4HH, UK.
Meczes
E L
EL
Perenna Rogers
M
M
Lock
R B
RB
Sullivan
D M
DM
Finlay
G J
GJ
Baguley
B C
BC
Austin
C A
CA
D000166:Acridines / Q000494:pharmacology; D000677:Amsacrine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D000951:Antigens, Neoplasm; D000970:Antineoplastic Agents / Q000494:pharmacology*; D016466:CHO Cells; D018827:Carcinoma, Lewis Lung / Q000188:drug therapy / Q000201:enzymology; D002453:Cell Cycle / Q000187:drug effects; D006224:Cricetinae; D004250:DNA Topoisomerases, Type II / Q000302:isolation & purification; D004268:DNA-Binding Proteins; D005047:Etoposide / Q000494:pharmacology; D006801:Humans; D007527:Isoenzymes / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D051379:Mice; D000075223:Poly-ADP-Ribose Binding Proteins; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000201:enzymology; D059005:Topoisomerase II Inhibitors; D014407:Tumor Cells, Cultured
D000166:Acridines; D000951:Antigens, Neoplasm; D000970:Antineoplastic Agents; C117791:Cl-DACA; D004268:DNA-Binding Proteins; D007527:Isoenzymes; D000075223:Poly-ADP-Ribose Binding Proteins; D059005:Topoisomerase II Inhibitors; C105581:methyl-N-(4-(9-acridinylamino)-2-methoxyphenyl)carbamate; D000677:Amsacrine; D005047:Etoposide; C058332:N-(2'-(dimethylamino)ethyl)acridine-4-carboxamide; D004250:DNA Topoisomerases, Type II; C000618811:TOP2A protein, human; C000618812:Top2a protein, mouse