Wright A W, Nocente M L, Smith M T
School of Pharmacy, The University of Queensland, Brisbane, Australia.
Life Sci. 1998;63(5):401-11. doi: 10.1016/s0024-3205(98)00288-4.
Hydromorphone-3-glucuronide (H3G) was synthesized biochemically using rat liver microsomes, uridine-5'-diphosphoglucuronic acid (UDPGA) and the substrate, hydromorphone. Initially, the crude putative H3G product was purified by ethyl acetate precipitation and washing with acetonitrile. Final purification was achieved using semi-preparative high-performance-liquid-chromatography (HPLC) with ultraviolet (UV) detection. The purity of the final H3G product was shown by HPLC with electrochemical and ultraviolet detection to be > 99.9% and it was produced in a yield of = 60% (on a molar basis). The chemical structure of the putative H3G was confirmed by enzymatic hydrolysis of the glucuronide moiety using beta-glucuronidase, producing a hydrolysis product with the same HPLC retention time as the hydromorphone reference standard. Using HPLC with tandem mass spectrometry (HPLC-MS-MS) in the positive ionization mode, the molecular mass (M+1) was found to be 462 g/mol, in agreement with H3G's expected molecular weight of 461 g/mol. Importantly, proton-NMR indicated that the glucuronide moiety was attached at the 3-phenolic position of hydromorphone. A preliminary evaluation of H3G's intrinsic pharmacological effects revealed that following i.c.v. administration to adult male Sprague-Dawley rats in a dose of 5 microg, H3G evoked a range of excitatory behavioural effects including chewing, rearing, myoclonus, ataxia and tonic-clonic convulsions, in a manner similar to that reported previously for the glucuronide metabolites of morphine, morphine-3-glucuronide and normorphine-3-glucuronide.
使用大鼠肝微粒体、尿苷-5'-二磷酸葡萄糖醛酸（UDPGA）和底物氢吗啡酮通过生化方法合成了氢吗啡酮-3-葡萄糖醛酸苷（H3G）。最初，通过乙酸乙酯沉淀并用乙腈洗涤对粗制的假定H3G产物进行纯化。使用具有紫外（UV）检测的半制备高效液相色谱（HPLC）实现最终纯化。通过具有电化学和紫外检测的HPLC显示，最终H3G产物的纯度>99.9%，并且其产率为60%（基于摩尔）。使用β-葡萄糖醛酸苷酶对葡萄糖醛酸苷部分进行酶促水解，产生一种水解产物，其HPLC保留时间与氢吗啡酮参考标准品相同，从而证实了假定H3G的化学结构。在正离子模式下使用HPLC串联质谱（HPLC-MS-MS），发现分子量（M+1）为462 g/mol，与H3G预期分子量461 g/mol一致。重要的是，质子核磁共振表明葡萄糖醛酸苷部分连接在氢吗啡酮的3-酚羟基位置。对H3G内在药理作用的初步评估显示，以5微克的剂量对成年雄性Sprague-Dawley大鼠进行脑室内给药后，H3G引发了一系列兴奋性行为效应，包括咀嚼、竖毛、肌阵挛、共济失调和强直性阵挛性惊厥，其方式与先前报道的吗啡的葡萄糖醛酸苷代谢物、吗啡-3-葡萄糖醛酸苷和去甲吗啡-3-葡萄糖醛酸苷相似。
