Abstract
Arylamine N-acetyltransferase 1 (NAT1) plays a pivotal role in the metabolism of carcinogens and is a drug target for cancer prevention and/or treatment. A protein–ligand virtual screening of 2 million chemicals was ranked for predicted binding affinity towards the inhibition of human NAT1. Sixty of the five hundred top-ranked compounds were tested experimentally for inhibition of recombinant human NAT1 and N-acetyltransferase 2 (NAT2). The most promising compound 9,10-dihydro-9,10-dioxo-1,2-anthracenediyl diethyl ester (compound 10) was found to be a potent and selective NAT1 inhibitor with an in vitro IC50 of 0.75 µM. Two structural analogs of this compound were selective but less potent for inhibition of NAT1 whereas a third structural analog 1,2-dihydroxyanthraquinone (a compound 10 hydrolysis product also known as Alizarin) showed comparable potency and efficacy for human NAT1 inhibition. Compound 10 inhibited N-acetylation of the arylamine carcinogen 4-aminobiphenyl (ABP) both in vitro and in DNA repair-deficient Chinese hamster ovary (CHO) cells in situ stably expressing human NAT1 and CYP1A1. Compound 10 and Alizarin effectively inhibited NAT1 in cryopreserved human hepatocytes whereas inhibition of NAT2 was not observed. Compound 10 caused concentration-dependent reductions in DNA adduct formation and DNA double-strand breaks following metabolism of aromatic amine carcinogens beta-naphthylamine and/or ABP in CHO cells. Compound 10 inhibited proliferation and invasion in human breast cancer cells and showed selectivity towards tumorigenic versus non-tumorigenic cells. In conclusion, our study identifies potent, selective, and efficacious inhibitors of human NAT1. Alizarin’s ability to inhibit NAT1 could reduce breast cancer metastasis particularly to bone.
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The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- NAT1:
-
Arylamine N-acetyltransferase 1
- NAT2:
-
Arylamine N-acetyltransferase 2
- PABA:
-
p-Aminobenzoic acid
- SMZ:
-
Sulfamethazine
- BNA:
-
Beta-naphthylamine
- ABP:
-
4-Aminobiphenyl
- CHO:
-
Chinese hamster ovary
- AcCoA:
-
Acetyl coenzyme A
- HPLC:
-
High performance liquid chromatography
- dG:
-
Deoxyguanosine
- N-OH-ABP:
-
N-Hydroxy-4-aminobiphenyl
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Acknowledgements
Portions of this work constitute partial fulfillment for the PhD in pharmacology and toxicology at the University of Louisville to Carmine S. Leggett. We thank Jason Walraven, a prior PhD graduate student in our laboratory, for his contributions towards screening the ZINC library.
Funding
This work was funded in part by NIH grants T32-ES011564, P20-RR18733, and P30-ES030283.
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CSL: conceptualization, methodology, validation, formal analysis, investigation, writing—original draft, writing—review & editing, visualization. MAD: formal analysis, investigation, writing—review & editing. RAS-G: formal analysis, investigation, writing—review & editing. MH: formal analysis, investigation, writing—review & editing. JOT: conceptualization, methodology, software, validation, formal analysis, resources, writing—review & editing. DWH: conceptualization, methodology, validation, formal analysis, resources, writing—review & editing, visualization, supervision.
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Leggett, C.S., Doll, M.A., Salazar-González, R.A. et al. Identification and characterization of potent, selective, and efficacious inhibitors of human arylamine N-acetyltransferase 1. Arch Toxicol 96, 511–524 (2022). https://doi.org/10.1007/s00204-021-03194-x
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DOI: https://doi.org/10.1007/s00204-021-03194-x