Modulation in kinetics of lactone ring hydrolysis of camptothecins upon interaction with topoisomerase I cleavage sites on DNA.
The kinetics of hydrolysis of the alpha-hydroxylactone ring of anticancer agents belonging to the camptothecin (CPT) series has been followed using their fluorescence emission. Data obtained for CPT, CPT-11, and SN-38, either in their free form or in the presence of DNA and/or topoisomerase I (top1), have been compared. DNA was modeled using three types of double-strand oligonucleotides corresponding to top1 cleavage site enhanced in the presence of the drug (olg1), top1 site independent of CPT (olg2), and nonspecific synthetic oligonucleotide containing only AT and no GC base pairs (olg3). Cleavage assays indicated the absence of top1-mediated cleavage on olg3, both in the presence and in the absence of CPT. The kinetics data also showed ratio-dependent stabilization of the lactone forms of CPTs when in the presence of an excess of olg1 or olg2, but not of olg3. These observations correlate with the previously reported preferential binding of CPTs to guanines. Although lactone hydrolysis was not perturbed by top1 alone, this enzyme hindered lactone stabilization by specific oligonucleotides. After addition of top1 to CPT-olg1 or CPT-olg2 complexes, the lactone ring of the drug was destabilized. No lactone stabilization was observed when olg1 was added to CPT-top1 complexes or when olg1-top1 complexes were added to CPT.
Chourpa
I
I
Laboratoire de Spectroscopie Biomoléculaire, UFR de Pharmacie, Université de Reims Champagne-Ardenne, France.
Riou
J F
JF
Millot
J M
JM
Pommier
Y
Y
Manfait
M
M
D002166:Camptothecin / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D004264:DNA Topoisomerases, Type I / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D006868:Hydrolysis / Q000187:drug effects; D000077146:Irinotecan; D007700:Kinetics; D007783:Lactones / Q000737:chemistry / Q000378:metabolism*; D009841:Oligonucleotides / Q000494:pharmacology; D013539:Simian virus 40 / Q000235:genetics; D013050:Spectrometry, Fluorescence; D059004:Topoisomerase I Inhibitors
D004279:DNA, Viral; D007783:Lactones; D009841:Oligonucleotides; D059004:Topoisomerase I Inhibitors; D000077146:Irinotecan; D004264:DNA Topoisomerases, Type I; D002166:Camptothecin