熊果酸前药的设计合成及抗多发性硬化症活性评价

Design, synthesis and Anti-multiple sclerosis activity evaluation of ursolic acid prodrugs

关键词:  熊果酸  前药  琥珀酸酯  多发性硬化症  结构改造

Key words:  ursolic acid  prodrug  succinate  multiple sclerosis  structural modification

摘要:

目的 设计、合成熊果酸前药，以改善熊果酸的水溶性，并对其抗多发性硬化症(multiple sclerosis，MS)活性进行评价。方法 根据前药策略，选取了琥珀酸、二肽氨基酸和艾莎康唑侧链片段，通过缩合、酯化反应等与熊果酸的3位羟基进行结合，采用紫外-可见分光光度法测定前药的水溶性，并建立小鼠实验性自身免疫性脑脊髓炎模型来进行抗MS活性评价。结果 共合成了3个前药，目标化合物的结构经¹H-NMR、¹³C-NMR和LCMS(ESI)进行确证；前药1和2水溶性较熊果酸提升110倍以上，前药3水溶性提升200倍以上；体内抗MS活性结果显示，所得前药化合物的抗MS活性均有一定提高，其中前药1的抗MS活性显著高于熊果酸组，其机制可能是通过抑制外周炎症细胞浸润中枢以及抗脱髓鞘从而发挥抗MS活性；前药1体内代谢结果显示，前药1以原型形式在体内的暴露量为熊果酸的6倍，表明前药1可能主要是以钠盐原型发挥抗MS活性。结论 前药1具有较大的抗MS潜力，值得深入研究。该研究为开发具有抗MS活性的熊果酸前药提供一定依据及有益指导。

Abstract:

OBJECTIVE Designing and synthesizing ursolic acid prodrugs to improve the water solubility and anti-multiple sclerosis(MS) activity of ursolic acid. METHODS succinic acid, dipeptide amino acid and the amino acids segments of side chains of isavuconazole wers selected according to the prodrug strategy, combined with 3-position hydroxyl of ursolic acid via condensation and esterification reaction. Using ultraviolet-visible spectrophotometry to determine the water solubility of prodrugs, and establish the experimental allergic encephalomyelitis mice model for its anti-MS efficacy evaluation. RESULTS Three prodrugs were synthesized and their structures were confirmed by ¹H-NMR, ¹³C-NMR and LCMS(ESI). The water solubility of prodrugs 1 and 2 was over 110 times higher than ursolic acid, and that of prodrug 3 was over 200 times higher than ursolic acid. The in vivo anti-MS activity results showed that the three prodrugs showed significant anti-MS activity, among which prodrug 1 showed significantly better anti-MS activity than ursolic acid group. The mechanism might be through inhibition of peripheral inflammatory cell infiltration into the center and anti-demyelination, so as to exert the anti-MS activity. The metabolism of prodrug 1 in vivo showed that the exposure of prodrug 1 in vivo was 6 times that of ursolic acid, indicating that prodrug 1 might exert anti-MS activity mainly as sodium prototype. CONCLUSION Prodrug 1 has great anti-MS potential, which is worthy of further study. This study provides some basis and useful guidance for the development of ursolic acid prodrugs with anti-MS activity.