Structure Elucidation of the Metabolites of 2', 3', 5'-Tri-O-Acetyl-N6-(3-Hydroxyphenyl) Adenosine in Rat Urine by HPLC-DAD, ESI-MS and Off-Line Microprobe NMR.
Structure Elucidation of the Metabolites of 2', 3', 5'-Tri-O-Acetyl-N6-(3-Hydroxyphenyl) Adenosine in Rat Urine by HPLC-DAD, ESI-MS and Off-Line Microprobe NMR.
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通过 HPLC-DAD、ESI-MS 和离线微探针 NMR 解析大鼠尿液中 2', 3', 5'-三-O-乙酰基-N6-(3-羟基苯基) 腺苷代谢物的结构。
DOI：
10.1371/journal.pone.0127583
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Wang Y
中科院分区：
文献类型：
--
作者：
Guo W;Jin M;Miao Z;Qu K;Liu X;Zhang P;Qin H;Zhu H;Wang Y
2‘，3’，5‘-三-O-乙酰基-N6-(3-羟基苯基)腺苷(又称WS070117)是一种新型的腺苷类似物，在体内和体外实验中均显示出一定的降血脂活性。由于其新的结构，人们对WS070117的代谢知之甚少。因此，我们研究了WS070117口服给药后大鼠尿液中的体内代谢产物。采用高效液相色谱-二极管阵列检测器(DAD)、离子俘获电喷雾电离-质谱仪(ESI-MS)和离线微探针核磁共振(NMR)联用技术对代谢产物进行鉴定。从亚毫克到毫克水平获得了七种纯化合物代谢物。结构分析结果表明，I相代谢产物N6-(3-羟基苯基)腺苷(M8)是WS070117在3个酯基上的水解物。N6-(3-羟基苯基)腺嘌呤(M7)也是I相代谢产物之一，是M8失去核呋喃糖后的衍生物。WS070117在大鼠尿液中除两种I相代谢产物外，还存在5种II相代谢产物。8-羟基-N6-(3-羟基苯基)腺苷(M6)是M7在第8位的水解产物，其余4个分别为N6-(3-O-β-D-葡萄糖醛酸苯基)腺嘌呤(M2)、N8-羟基-N6-(3-O-磺基苯基)腺嘌呤(M3)、N6-(3-O-β-D-葡萄糖醛酸苯基)腺苷(M4)和N6-(3-O-磺基苯基)腺苷(M5)。第二阶段的代谢途径被证明包括羟基化、葡萄糖醛酸化和硫酸盐化。这项研究为WS070117的代谢提供了新的有价值的信息，也证明了高效液相/质谱学/离线微探针核磁共振方法是一种快速鉴定代谢物的可靠手段。
2', 3', 5'-tri-O-acetyl-N6-(3-hydroxyphenyl) adenosine (also known as WS070117) is a new adenosine analog that displays anti-hyperlipidemic activity both in vitro and in vivo experiments as shown in many preliminary studies. Due to its new structure, little is known about the metabolism of WS070117. Hence, the in vivo metabolites of WS070117 in rat urine following oral administration were investigated. Identification of the metabolites was conducted using the combination of high-performance liquid chromatography (HPLC) coupled with diode array detector (DAD), ion trap electrospray ionization-mass spectrometry (ESI-MS), and off-line microprobe nuclear magnetic resonance (NMR) measurements. Seven metabolites were obtained as pure compounds at the sub-milligram to milligram levels. Results of structure elucidation unambiguously revealed that the phase I metabolite, N6-(3-hydroxyphenyl) adenosine (M8), was a hydrolysate of WS070117 by hydrolysis on the three ester groups. N6-(3-hydr-oxyphenyl) adenine (M7), also one of the phase I metabolites, was the derivative of M8 by the loss of ribofuranose. In addition to two phase I metabolites, there were five phase II metabolites of WS070117 found in rat urine. 8-hydroxy-N6-(3-hydroxy-phenyl) adenosine (M6) was the product of M7 by hydrolysis at position 8. The other four were elucidated to be N6-(3-O-β-D-glucuronyphenyl) adenine (M2), N8-hydroxy-N6-(3-O-sulfophenyl) adenine (M3), N6-(3-O-β-D-glucuronyphenyl) adenosine (M4), and N6-(3-O- sulfophenyl) adenosine (M5). Phase II metabolic pathways were proven to consist of hydroxylation, glucuronidation and sulfation. This study provides new and valuable information on the metabolism of WS070117, and also demonstrates the HPLC/MS/off-line microprobe NMR approach as a robust means for rapid identification of metabolites.
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影响因子：
1.8
作者：
Liu, Yin;He, Jiuming;Zhu, Haibo
通讯作者：
Zhu, Haibo
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2.8
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Simons, LA
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Simons, LA
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Griffin, JL;Nicholls, AW;Kuehn, T
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Schmidt, Bonnie;Jaroszewski, Jerzy W.;Staerk, Dan
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1.8
作者：
Nicholson, JK;Lindon, JC;Holmes, E
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Holmes, E