Inhibition of RNA synthesis in vitro and cell growth by anthracycline antibiotics.
New derivatives of doxorubicin and daunorubicin with amidine group bonded to daunosamine at C-3' atom and bearing the morpholine ring attached to the amidine group have been recently synthesized. Their cytotoxic activities and effects on RNA synthesis in vitro were assayed. The drug concentrations inhibiting mouse leukaemia L1210 cell growth to 50% were about two- and three fold higher for the derivatives compared to doxorubicin and daunorubicin respectively. Inhibition of phage T7 RNA polymerase by the non-covalently interacting derivatives was also slightly lower than that by the parent compounds. As doxorubicin and daunorubicin, their amidine derivatives in the presence of dithiothreitol and Fe(III) ions are activated and covalently bind to DNA. The adducts formed affect RNA polymerase activity. Several bands corresponding to prematurely terminated RNA chains are observed by means of polyacrylamide gel electrophoresis. The patterns of bands are virtually identical for all the anthracyclines studied here and are similar to the terminations induced by actinomycin D. This observation is consistent with a notion that the adducts are formed at guanine in GpC sequences which are also binding sites of actinomycin D. A substantial difference between daunorubicin and its amidine derivative is shown by means of high performance liquid chromatography. The derivative undergoes rapid rearrangements in the presence of dithiothreitol and Fe(III) ions, while daunorubicin is stable for several hours under these conditions. The results presented here indicate that the amidine derivatives despite bulky morpholine substitution exhibit biological activity in the systems used here.
Studzian
K
K
Department of General Chemistry, Institute of Physiology and Biochemistry, Medical University of Lódź, Poland. magn@csk.am.lodz.pl
Wasowska
M
M
Piestrzeniewicz
M K
MK
Wilmańska
D
D
Szmigiero
L
L
Oszczapowicz
I
I
Gniazdowski
M
M
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D002455:Cell Division; D002851:Chromatography, High Pressure Liquid; D018899:CpG Islands; D018736:DNA Adducts; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D003609:Dactinomycin / Q000494:pharmacology; D003630:Daunorubicin / Q000031:analogs & derivatives*; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000031:analogs & derivatives*; D051379:Mice; D008956:Models, Chemical; D010957:Plasmids / Q000378:metabolism; D012313:RNA / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000378:metabolism; D013696:Temperature; D013997:Time Factors; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D000903:Antibiotics, Antineoplastic; D018736:DNA Adducts; D012333:RNA, Messenger; D003609:Dactinomycin; D012313:RNA; D004317:Doxorubicin; D012321:DNA-Directed RNA Polymerases; D003630:Daunorubicin