目的 制备葫芦素B磷脂复合物(CuB-PLC),对其进行理化性质评价和体外抗肿瘤活性测定.方法 采用溶剂挥发法制备CuB-PLC,用Box-Behnken设计-效应面法优化其制备工艺,考察其表观油水分配系数、粒径及形态;用X射线衍射光谱(XRD)、红外光谱(IR)综合分析CuB-PLC的形成机制,利用MTT法测定其体外抗肿瘤活性.结果 优化的CuB-PLC制备工艺为以四氢呋喃作为反应溶剂,磷脂与葫芦素B的投药物质的量比为1:1,反应质量浓度为1.5 mg/mL,反应温度和反应时间分别为60 ℃和3 h,复合率可达97.15%.XRD和IR分析表明葫芦素B与磷脂未形成新的化合物或混合物,而是磷脂复合物;CuB-PLC在水中的溶解度提高到原料药的3.9倍;在水中复合物粒径为(521.30±10.50)nm,多分散指数(PDI)为0.133 2±0.024 0;MTT实验表明,葫芦素B与CuB-PLC对HepG2细胞增殖的半数抑制浓度(IC₅₀)值分别为42.55、27.61 μmol/L.结论 成功制备了CuB-PLC,工艺简单可行,复合率高,水溶性有所改善,且对HepG2细胞增殖的抑制作用显著增强,为葫芦素B的制剂学研究提供了参考.

Objective To prepare cucurbitacin B phospholipids complex (CuB-PLC) and evaluate its physicochemical properties and in vitro antitumor activity. Methods CuB-PLC was prepared using solvent evaporation method and optimized by Box-Behnken design. The oil-water partition coefficient, particle size, and morphology of CuB-PLC were investigated; X-ray diffraction (XRD) spectroscopy and infrared (IR) spectroscopy were used to analyze the formation machenism of CuB-PLC. MTT method was used to determine the in vitro antitumor activity of CuB-PLC. Results The optimal formulation protocol for CuB-PLC was as follows: Tetrahydrofuran was taken as the reaction medium, phospholipids-cucurbitacin B molar ratio, reaction concentration of cucurbitacin B, reaction temperature and time were 1:1, 1.5 mg/mL, 60 ℃, and 3 h, respectively. The complex rate and particle size for the optimized CuB-PLC was 97.15% and (521.30 ± 10.50) nm, and the polydispersity index (PDI) was 0.133 2 ± 0.024 0. MTT experiments showed that the half of the HepG-2 cell proliferation inhibition concentration (IC₅₀) values of CuB and CuB-PLC were 42.55 and 27.61 μmol/L. Conclusion CuB-PLC is successfully developed under the optimized protocol, possessing high complex rate, and enhanced solubility in water, and the inhibition on HepG-2 cell proliferation is significantly enhanced, which provides the reference for the further research of CuB.

北京市科委重点项目(Z131100002513005);国家教育部留学归国人员科研启动基金(20101561)