Chiba T, Nabeshima S, Takei Y, Onozaki K
Department of Hygienic Chemistry, Faculty of Pharmaceutical Sciences, Nagoya City University, Tanabe, Mizuho, Japan.
Glycoconj J. 1998 Jan;15(1):63-7. doi: 10.1023/a:1006991416735.
In order to develop glycosylated cytokine, recombinant human IL-1alpha was chemically modified with galactose monosaccharide. Galactose with C9 spacer, 8-(hydrazinocarbonyl)octyl beta-D-galactopyranoside (3), was synthesized by glycosylation of C9 spacer, methyl 9-hydroxynonanoate, with acetobromogalactose, followed by deacetylation and hydrazidation. Total yield of 3 was 43.6% in three steps. Compound 3 was coupled to IL-1alpha by the acyl azide method. The glycosylated IL-1 was purified by anion-exchange chromatography, and galactose coupled to IL-1 was confirmed by R. communis lectin blotting. Based on the molecular weight, the average number of carbohydrate molecules introduced per molecule of IL-1alpha was estimated to be 9.1.
为了开发糖基化细胞因子，重组人白细胞介素-1α（IL-1α）用半乳糖单糖进行了化学修饰。带有C9间隔基的半乳糖，8-(肼羰基)辛基β-D-吡喃半乳糖苷(3)，通过C9间隔基9-羟基壬酸甲酯与乙酰溴半乳糖进行糖基化反应，随后进行脱乙酰化和肼化反应合成。三步反应中3的总产率为43.6%。化合物3通过酰基叠氮法与IL-1α偶联。糖基化的IL-1通过阴离子交换色谱法纯化，并且通过蓖麻凝集素印迹法确认了与IL-1偶联的半乳糖。根据分子量，估计每分子IL-1α引入的碳水化合物分子平均数量为9.1。
