目的 合成一种雷公藤红素-氨基葡萄糖酰胺偶联物（celastrol-glucosamine conjugate，Cel-GlcN），并评估其细胞毒性、调脂活性，以缓解Cel不良反应强、水溶性差的问题。方法 采用氨基葡萄糖对Cel第20位的羧酸基团进行酰胺化修饰，分别在二氯甲烷、N,N-二甲基甲酰胺（N,N-dimethylformamide，DMF）体系中使用1-乙基-(3-二甲基氨基丙基)碳酰二亚胺［1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride，EDC］、六氟磷酸苯并三唑-1-基-氧基三吡咯烷基磷（benzotriazol-1-yl-oxytripyrrolidino-phosphonium hexafluorophosphate，PyBOP）作为酰胺偶联反应的活化剂加速Cel-GlcN的合成，使用高效液相色谱-质谱（HPLC-MS/MS）、¹H-NMR、¹³C-NMR和傅里叶变换红外光谱（Fourier transform infrared spectroscopy，FTIR）对其结构进行了表征，基于人肝癌HepG2细胞模型评价其细胞毒性、调脂活性及氧化应激水平。结果 FTIR确定了化合物中酰胺键的存在，HPLC-MS/MS测定了化合物的相对分子质量，¹H-NMR、¹³C-NMR鉴定了化合物的具体化学结构，相比于天然产物Cel，Cel-GlcN的水溶性得到了显著升高（由常温下的1.23 μg/mL提升为226.00 μg/mL），可能以缓控释作用释放Cel从而减轻其对细胞的毒性和机体内的氧化应激水平。结论 成功合成了Cel-GlcN，水溶性得到改善，体外细胞实验证实其调脂活性基本不变而细胞毒性降低，从而为Cel相关药物的研发提供有价值的参考。

Objective To synthesize a celastrol-glucosamine conjugate (Cel-GlcN) and evaluate its cytotoxicity and lipid-lowering activity in an effort to mitigate the strong side effects and poor water solubility of Cel.Methods The carboxylic acid group at the 20th position of Cel was aminated with glucosamine. The synthesis process was expedited utilizing 1-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (EDC) and benzotriazol-1-yl-oxytripyrrolidino-phosphonium hexafluorophosphate (PyBOP) as conjugating agents in dichloromethane and N,N-dimethylformamide (DMF) systems. The structure of the derivative was further characterized through HPLC-MS, ¹H-NMR, ¹³C-NMR, and Fourier transform infrared spectroscopy (FTIR). The cytotoxicity, lipid-lowering effect, and oxidative stress level of the derivative were evaluated using the HepG2 cell model. Results The presence of amide bond(s) in the compound was confirmed through FTIR analysis. Furthermore, the relative molecular weight was determined using HPLC-MS/MS, and the specific chemical structure of the derivative was identified as Cel-GlcN through ¹H NMR and ¹³C NMR spectroscopy. Compared to the natural product Cel, the water solubility of Cel-GlcN significantly improved to 226 μg/mL. This improvement suggests potential mitigation of cytotoxicity and reduction of oxidative stress levels in the body via controlled release of Cel.Conclusion The successful synthesis of Cel-GlcN was accomplished, leading to enhanced water solubility. In vitro cellular experiments verified the retained lipid-lowering activity and a decrease in cytotoxicity. These findings provide significant insights for the development of drugs related to Cel.

国家科技重大专项子课题（2018ZX0921002-008）