Genotoxicity of 5-hydroxymethylfurfural and its dipolymer and metabolite

doi:10.19803/j.1672-8629.20210706

Abstract

Abstract: Objective To evaluate the potential genotoxicity of impurities-5-hydroxymethylfurfural (5-HMF), its polymer 5,5’-oxydimethylenebis(2-furfural) (OMBF), and its metabolite 5-sulfooxymethylfurfural (5-SMF). Methods The genotoxicity of 5-HMF, OMBF and 5-SMF was predicted by means of Derek and Sarah with the quantitative structure-activity relationship (QSAR) technology. The bacterial reverse-mutation (Ames) test and single cell gel electrophoresis assay (SCGE, also called the comet assay) were performed to verify the above results. In Ames tests, the potential mutagenicity was evaluated using six strains of S. typhimurium (TA97、TA98、TA100、TA102、TA1535 and TA1537) treated with five different concentrations of 5-HMF, OMBF and 5-SMF with or without S9 mix from liver microsomes of mice, rats or humans and incubated for 48 h. DMSO 20 μg/well served as the vehicle control. In the absence of S9 mix, AF-2, NaN3 and 9-AA served as the positive control, otherwise 2-AA served as the positive control. When TA1535 and TA1537 had three times as many colonies as the vehicle control and when the number of colonies of TA97, TA98, TA100 and TA102 was twice that of the vehicle control, the test compound was considered mutagenic. Results In silico evaluation by Derek software showed that 5-HMF and 5-SMF were equivocally positive mutagenic and OMBF was inactive, while Sarah software predicted that 5-HMF and 5-SMF were positive mutagenic and OMBF was equivocally positive mutagenic. Ames test results demonstrated that 5-HMF and OMBF at a dose greater than 750 μg/well increased the number of TA100 revertant colonies only with rat-liver S9 mix. 5-SMF was positive mutagenic with mouse-, rat- or human-liver S9 mix on TA97, TA100, TA102 and TA1535 at a dose above 20~40 μg·well^-1, or without S9 mix on TA97, TA98, TA100 and TA102 at a dose over 50 μg·well^-1. Conclusion 5-HMF and OMBF show potential mutagenicity with a rat-liver S9 mix, while 5-SMF shows mutagenicity with/without S9 mix in vitro. Therefore, 5-HMF, OMBF and 5-SMF are class 2 genotoxic impurities, thus the limit of the three impurities should be enforced to ensure the safety of drugs, especially for people who may be chronically exposed to any of them.

Key words: 5-hydroxymethylfurfural, 5,5'-oxydimethylenebis(2-furfural), 5-sulfooxymethylfurfural, genotoxic impurity, (quantitative) structure-activity relationship, mutagenicity, Ames test

CLC Number:

R991
R994.11

LIN Ni, YE Qian, GENG Xingchao, WANG Xue, JIN Hongtao, WEN Hairuo. Genotoxicity of 5-hydroxymethylfurfural and its dipolymer and metabolite[J]. Chinese Journal of Pharmacovigilance, 2023, 20(2): 157-162.

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