Profiling and identification of metabolites of antisense oligonucleotides AD00510-AS in liver S9 and plasma of mouse,rat,monkey,and human
Objective:To establish an LC-UV-Q-TOF method of the antisense oligonucleotides(ASO)drug AD00510-AS as well as its metabolites in liver S9 and plasma,then propose the in vitro metabolic pathways of AD00510-AS.Methods:After 24 h of incubation,samples of liver S9 and plasma were cleaned up by solid phase extraction.Data was acquired with SCIEX OS software and processed with Molecule ProfilerTM software.Metabolites were identified by comparing extracted ion chromatograms(XICs)between control and analysis samples,then metabolic pathways were proposed.Results:Totally three metabolites(M1,M4 and M5)of AD00510-AS were detected and tentatively identified in mouse,rat,monkey,and human liver S9;the AD00510-AS accounted for 73.9%,87.1%,78.4%and 81.3%of the total drug-related components in mouse,rat,monkey,and human liver S9,respectively.Totally four metabolites(M2,M3,M4 and M5)of AD00510-AS were detected and tentatively identified in mouse,rat,monkey,and human plasma;the AD00510-AS accounted for 69.1%,94.5%,67.8%and 68.1%of the total drug-related components in mouse,rat,monkey,and human plasma,respectively.Based on the MS and MS/MS fragment ions measurement,the identified metabolites were as follows:M1 was a metabolite(MW =4 544)after 9 nucleotides lost from the 3'end of AD00510-AS,M2 was a metabolite(MW =6857)after 2 nucleotides lost from the 3'end of AD00510-AS,M3 was a metabolite(MW =7232)after 1 nucleotide lost from the 3'end of AD00510-AS,M4 was a di-desulfurization metabolite(MW =7 559),and M5 was a desulfurization metabolite(MW = 7 575).Conclusion:AD00510-AS and its five metabolites were detected and identified by validated LC-UV-Q-TOF method from liver S9 and plasma;AD00510-AS is mainly metabolized via degradation(loss of nucleotides)and desulfurization.
antisense oligonucleotidesliver S9plasmametabolite(s)profiling and identificationsolid phase extraction
NETL
NSTL
万方数据
维普
