Identification and characterization of human apurinic/apyrimidinic endonuclease-1 inhibitors.
The repair of abasic sites that arise in DNA from hydrolytic depurination/depyrimidination of the nitrogenous bases from the sugar-phosphate backbone and the action of DNA glycosylases on deaminated, oxidized, and alkylated bases are critical to cell survival. Apurinic/apyrimidinic endonuclease-1/redox effector factor-1 (APE-1; aka APE1/ref-1) is responsible for the initial removal of abasic lesions as part of the base excision repair pathway. Deletion of APE-1 activity is embryonic lethal in animals and is lethal in cells. Potential inhibitors of the repair function of APE-1 were identified based upon molecular modeling of the crystal structure of the APE-1 protein. We describe the characterization of several unique nanomolar inhibitors using two complementary biochemical screens. The most active molecules all contain a 2-methyl-4-amino-6,7-dioxolo-quinoline structure that is predicted from the modeling to anchor the compounds in the endonuclease site of the protein. The mechanism of action of the selected compounds was probed by fluorescence and competition studies, which indicate, in a specific case, direct interaction between the inhibitor and the active site of the protein. It is demonstrated that the inhibitors induce time-dependent increases in the accumulation of abasic sites in cells at levels that correlate with their potency to inhibit APE-1 endonuclease excision. The inhibitor molecules also potentiate by 5-fold the toxicity of a DNA methylating agent that creates abasic sites. The molecules represent a new class of APE-1 inhibitors that can be used to probe the biology of this critical enzyme and to sensitize resistant tumor cells to the cytotoxicity of clinically used DNA damaging anticancer drugs.
Department of Pharmaceutical Sciences, University of Pittsburgh, Pittsburgh, PA 15261, USA.
Ajay
A
Srinivasan
Lirong
L
Wang
Cari J
CJ
Cline
Zhaojun
Z
Xie
Robert W
RW
Sobol
Xiang-Qun
XQ
Xie
Barry
B
Gold
D001483:Base Sequence; D020134:Catalytic Domain; D045744:Cell Line, Tumor; D004247:DNA / Q000235:genetics / Q000378:metabolism; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000378:metabolism / Q000494:pharmacology* / Q000633:toxicity; D006801:Humans; D062105:Molecular Docking Simulation; D010084:Oxidation-Reduction / Q000187:drug effects; D054852:Small Molecule Libraries / Q000378:metabolism / Q000494:pharmacology / Q000633:toxicity
D004791:Enzyme Inhibitors; D054852:Small Molecule Libraries; D004247:DNA; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase