反义寡核苷酸药物AD00510-AS在小鼠、大鼠、猴、人肝S9和血浆中的代谢产物研究  

Profiling and identification of metabolites of antisense oligonucleotides AD00510-AS in liver S9 and plasma of mouse,rat,monkey,and human

作　　者：柳庆龙 纪海霞 李伦 张云 祁佳琦 刘添浩 梁柳艺 陶桂勇 周绍联 LIU Qing-long;JI Hai-xia;LI Lun;ZHANG Yun;QI Jia-qi;LIU Tian-hao;LIANG Liu-yi;TAO Cui-yong;ZHOU Shao-lan(InnoStar Bio-tech Nantong Co.,Ltd.,Nantong 226133,China;Shanghai InnoStar Bio-tech Co.,Ltd.,Shanghai 201203,China;Shanghai Argo Biopharmaceutical Co.,Ltd.,Shanghai 201203,China)

机构地区：[1]益诺思生物技术南通有限公司,南通226133 [2]上海益诺思生物技术股份有限公司,上海201203 [3]上海舶望制药有限公司,上海201203

出　　处：《中国新药杂志》2024年第2期151-158,共8页Chinese Journal of New Drugs

基　　金：江苏省新药一站式高效非临床评价公共服务平台建设项目(BM2021002)。

摘　　要：目的:建立反义寡核酸药物AD00510-AS的LC-UV-Q-TOF方法,并探索小核酸药物AD00510-AS在体外肝S9和血浆中的代谢产物和代谢途径.方法:基于固相萃取的方法对孵育24h后的肝S9和血浆样品进行处理,采用SCIEX OS进行数据采集,采用Molecule ProfilerTM软件分析空白样品和给药后孵育24h样品的提取离子流图(XIC)的差异,得到可能的代谢产物,并推测代谢途径.结果:AD00510-AS分别与小鼠、大鼠、猴、人肝S9孵育24h后,共检测到3个代谢产物(M1,M4和M5),母药的相对丰度分别为73.9%,87.1%,78.4%和81.3%;AD00510-AS分别与小鼠、大鼠、猴、人血浆孵育24h后,共检测到4个代谢产物(M2,M3,M4和M5),母药的相对丰度分别为69.1%,94.5%,67.8%和68.1%;通过MS及MS/MS的测定,鉴定的代谢产物如下:M1:3'末端丢失9个核苷酸代谢产物(MW=4544);M2:3'末端丢失2个核苷酸代谢产物(MW=6857);M3:3'末端丢失1个核苷酸代谢产物(MW=7232);M4:双脱硫代谢产物(MW=7559);M5:脱硫代谢产物(MW=7575).结论:采用确认的LC-UV-Q-TOF方法可以检测到AD00510-AS及其5个代谢产物,AD00510-AS在体外肝S9和血浆中主要通过降解(丢失核苷酸)和脱硫进行代谢.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.

关 键 词：反义寡核苷酸 肝S9 血浆 代谢产物鉴定 固相萃取 

分 类 号：R969.1[医药卫生—药理学]