Determination and measurement of unusual metabolites of bile acid multi-conjugates excreted in a patient with Nimann-Pick disease type C1.

本文描述了3β，7β-二羟基-5-胆碱-24-酸的化学合成，硫酸在C-3， n-乙酰氨基葡萄糖（GlcNAc）在C-7和甘氨酸（或牛磺酸）在C-24，这是罕见的，主要代谢物排泄的尼曼-匹克病C1型患者的尿液。还制备了3β-羟基-7-氧-5-胆碱-24-酸的类似双共轭物。涉及的主要反应有：1)在煮沸的甲苯中，在CdCO_3存在下，3 - β-叔丁基二甲基硅氧烷(TBDMSi)-7 - β-羟基-5-胆碱-24-oate在C-7处与2-乙酰氨基-1α-氯-1,2-二脱氧-3,4,6-三-o -乙酰-d -葡萄糖吡喃糖在C-3处与三氧化硫-三甲胺配合物在吡啶中磺化生成3 - β-TBDMSi-7 - β- glcnac。3)在DMF中使用4-（4,6-二甲氧基-1,3,5-三嗪-2-基）-4-甲基酰氯作为偶联剂，在3β-磺基-7β- glcnac偶联物的C-24位点与甘氨酸甲酯盐酸盐（或牛磺酸）直接氨基酰化。采用液相色谱-质谱联用电喷雾探针对多共轭胆汁酸在正、负电离模式下的结构进行了表征。

The chemical synthesis of 3β,7β-dihydroxy-5-cholen-24-oic acid triply conjugated by sulfuric acid at C-3,N-acetylglucosamine (GlcNAc) at C-7 and glycine (or taurine) at C-24, which are unusual, major metabolites excreted in urine of a patient with Niemann-Pick diseases type C1,is described. Analogous double-conjugates of 3β-hydroxy-7-oxo-5-cholen-24-oic acid were also prepared. The principal reactions involved were : 1)β-D-N-acetylglueosaminidation at C-7 of methyl 3β-tert-butyldimethylsilyloxy (TBDMSi)-7β-hydroxy-5-cholen-24-oate with 2-acetamido-1α-chloro-1,2-dideoxy-3,4,6-tri-O-acetyl-D-glucopyranose in the presence of CdCO_3 in boiling toluene, 2)sulfation at C-3 of the resulting 3β-TBDMSi-7β-GlcNAc with sulfur trioxide-trimethylamine complex in pyridine, and 3)direct aminoacyl amidation at C-24 of the 3β-sulfo-7β-GlcNAc conjugate with glycine methyl ester hydrochloride (or taurine) using 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride as a coupling agent in DMF. The structures of the multi-conjugated bile acids were characterized by liquid chromatography-mass spectrometry with electrospray ionization probe under the positive and the negative ionization modes.