A sensitive liquid chromatography/electrospray tandem mass spectroscopy method for simultaneous quantification of a disulfide bond doxorubicin conjugation prodrug and activated doxorubicin:application to cellular pharmacokinetic and catabolism studies
In recent years,drug conjugates as a prodrug strategy have been widely studied,especially combined with nanotechnology.Disulfide-linked doxorubicin drug-drug conjugate(DOX-S-S-DOX)nanoparticles,have recently been developed as a doxorubicin prodrug nanoparticles with greater anticancer activity and less toxicity than doxorubicin in vivo,while its intracellular kinetics and metabolism is unclear which may provide us with a deeper understanding of its pharmacological mechanism and antitumor effect.Hence,in this study,a rapid and sensitive ultra high-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method was developed to detect doxorubicin(DOX)activated from DOX-S-S-DOX,as well as the prodrug itself in human breast cancer tumor cells(MCF-7).Sample preparation involved acetonitrile precipitation to extract the analytes simultaneously and bath sonication to remove intercalated DOX from DNA.The calibration range was 3-60 ng/mL for DOX and 20-400 ng/mL for DOX-S-S-DOX with the correlation coefficients(r2)≥ 0.99,using daunorubicin as internal standard(IS).The inter-and intra-assay precision(relative standard deviation,RSD%)of quality control samples was in the acceptable range(<15%)and relative error(RE%)for accuracy was between-5.35 and 9.18%for all analytes.Recovery(59.28-69.53%for DOX-S-S-DOX and 99.13-100.10%for DOX)and matrix effect(99.69-111.19%)was consistent,precise,and reproducible at different quality control levels in accordance with FDA guidance.Stability studies showed that DOX-S-S-DOX was unstable both during the bench-top and long-term storage,while the stability during sample preparation and LC-MS runtime was suitable for all the analytes.Hence,the samples should be prepared as soon as possible at the time point to prevent the catabolism of DOX-S-S-DOX.The assay was successfully used in the cellular metabolism and pharmacokinetics study of DOX-S-S-DOX and it may give a clue to explore analytical methods of other prodrug forms of DOX.
Doxorubicin prodrug LC-MS/MS Protein precipitation combined with sonication MCF-7 cells Pharmacokinetics and metabolism
Nan Zheng Xing Wang Yaoqi Wang Guobing Xu Hua Zhang Wenbing Dai Bing He Qiang Zhang Jiafu Ji Xueqing Wang
Key laboratory of Carcinogenesis and Translational Research(Ministry of Education/Beijing),National Beijing Key Laboratory of Molecular Pharmaceutics and New Drug Delivery Systems,School of Pharmaceut Key laboratory of Carcinogenesis and Translational Research(Ministry of Education/Beijing),National
国内会议
西安
英文
1470-1482
2017-11-04（万方平台首次上网日期，不代表论文的发表时间）