Rashid Md Mamunur, Oh Hyun-A, Lee Hyunbeom, Jung Byung Hwa
Molecular Recognition Research Center, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea; Department of Biological Chemistry, Korea University of Science and Technology (UST), Daejeon 34113, Republic of Korea.
Molecular Recognition Research Center, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea; Department of Biological Sciences, Konkuk University, Seoul 143-701, Republic of Korea.
J Pharm Biomed Anal. 2017 Oct 25;145:473-481. doi: 10.1016/j.jpba.2017.06.059. Epub 2017 Jul 8.
AZD8055 is an ATP-competitive specific dual mTOR inhibitor and exhibited potent antitumor activity on several types of solid tumors. However, the metabolism of AZD8055 in the body still remains unknown. In this study, metabolite identification of AZD8055 was performed using ultra high-performance liquid chromatography-ion trap mass spectrometry (UHPLC-IT-MS) through both in vitro and in vivo approaches using rat liver microsomes (RLMs) and rat plasma, urine and feces, respectively. A total of eight putative metabolites (five phase I and three phase II) were identified, and a tentative metabolic pathway was suggested for the first time. Considering the accurate mass and mass fragmentations of the detected metabolites, their plausible structures were suggested. Demethylation, hydroxylation, oxidation and morpholine ring opening were the major biotransformation processes for the phase-I metabolism, while phase-II metabolites were merely generated by the glucuronide conjugation reaction. The cumulative excretion of AZD8055 in urine and feces was 0.13% and 1.11% of the dose, respectively. When the semi-quantitative analysis of the metabolites was performed using UHPLC-MS/MS (ultra-performance liquid chromatography tandem mass spectrometry) to evaluate the overall trend of metabolites formation and excretion, AZD8055 was excreted more in the form of the metabolites than itself and their formation was very fast. Therefore it was presumed that biotransformation was playing a crucial role in its elimination. Ultimately, this study provides novel insights regarding the in vitro and in vivo biotransformations of AZD8055. Further investigations of metabolites of this potent anti-cancer compound could be beneficial for the antitumor drug design and development process.
AZD8055是一种ATP竞争性特异性双mTOR抑制剂，对多种实体瘤显示出强大的抗肿瘤活性。然而，AZD8055在体内的代谢情况仍不清楚。在本研究中，分别使用大鼠肝微粒体（RLM）以及大鼠血浆、尿液和粪便，通过体外和体内方法，采用超高效液相色谱-离子阱质谱（UHPLC-IT-MS）对AZD8055进行代谢物鉴定。共鉴定出8种推定代谢物（5种I相代谢物和3种II相代谢物），并首次提出了初步的代谢途径。根据检测到的代谢物的精确质量和质谱碎裂情况，推测了它们可能的结构。去甲基化、羟基化、氧化和吗啉环开环是I相代谢的主要生物转化过程，而II相代谢物仅通过葡萄糖醛酸结合反应生成。AZD8055在尿液和粪便中的累积排泄量分别为给药剂量的0.13%和1.11%。当使用UHPLC-MS/MS（超高效液相色谱串联质谱）对代谢物进行半定量分析以评估代谢物形成和排泄的总体趋势时，AZD8055以代谢物形式排泄的量比其本身更多，且它们的形成非常迅速。因此推测生物转化在其消除过程中起关键作用。最终，本研究为AZD8055的体外和体内生物转化提供了新的见解。对这种强效抗癌化合物的代谢物进行进一步研究可能有助于抗肿瘤药物的设计和开发过程。
