DNA interaction of the tyrosine protein kinase inhibitor PD153035 and its N-methyl analogue.
The brominated anilinoquinazoline derivative PD153035 exhibits a very high affinity and selectivity for the epidermal growth factor receptor tyrosine kinase (EGF-R TK) and shows a remarkable cytotoxicity against several types of tumor cell lines. In contrast, its N-methyl derivative, designated EBE-A22, has no effect on EGF-R TK but maintains a high cytotoxic profile. The present study was performed to explore the possibility that PD153035 and its N-methyl analogue might interact with double-stranded DNA, which is a primary target for many conventional antitumor agents. We studied the strength and mode of binding to DNA of PD153035 and EBE-A22 by means of absorption, fluorescence, and circular and linear dichroism as well as by a relaxation assay using human DNA topoisomerases. The results of various optical and gel electrophoresis techniques converge to show that both drugs bind to DNA and behave as typical intercalating agents. In particular, EBE-A22 unwinds supercoiled plasmid, stabilizes duplex DNA against heat denaturation, and produces negative CD and ELD signals, as expected for an intercalating agent. Extensive DNase I footprinting experiments performed with a large range of DNA substrates show that EBE-A22, but not PD153035, interacts preferentially with GC-rich sequences and discriminates against homooligomeric runs of A and T which are often cut more readily by the enzyme in the presence of the drug compared to the control. Altogether, the results provide the first experimental evidence that DNA is a target of anilinoquinazoline derivatives and suggest that this N-methylated ring system is a valid candidate for the development of DNA-targeted cytotoxic compounds. The possible relevance of selective DNA binding to activity is considered. The unexpected GC-selective binding properties of EBE-A22 entreat further exploration into the use of N-methylanilinoquinazoline derivatives as tools for designing sequence-specific DNA binding ligands.
Goossens
J F
JF
Laboratoire de Chimie Analytique, Faculté des Sciences Pharmaceutiques et Biologiques, Université de Lille 2, 59006 Lille, France.
Bouey-Bencteux
E
E
Houssin
R
R
Hénichart
J P
JP
Colson
P
P
Houssier
C
C
Laine
W
W
Baldeyrou
B
B
Bailly
C
C
D000970:Antineoplastic Agents / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D002942:Circular Dichroism; D004247:DNA / Q000378:metabolism*; D018983:DNA Footprinting; D004278:DNA, Superhelical / Q000378:metabolism; D003720:Densitometry; D004587:Electrophoresis, Agar Gel; D004791:Enzyme Inhibitors / Q000378:metabolism*; D066246:ErbB Receptors / Q000037:antagonists & inhibitors*; D006801:Humans; D007364:Intercalating Agents / Q000378:metabolism*; D008969:Molecular Sequence Data; D011489:Protein Denaturation; D011799:Quinazolines / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
D000970:Antineoplastic Agents; D004278:DNA, Superhelical; D004791:Enzyme Inhibitors; D007364:Intercalating Agents; D011799:Quinazolines; D004247:DNA; D066246:ErbB Receptors; C088860:4-((3-bromophenyl)amino)-6,7-dimethoxyquinazoline