摘要
目的探索一步合成马钱子碱氮氧化物(BNO)的工艺条件,并观察其体外抑瘤作用。方法以马钱子碱和双氧水(H2O2)为原料,采用氮氧化法一步合成BNO,运用正交试验设计研究原料配比、反应温度和溶剂(无水乙醇)体积对反应产率和目标产物纯度的影响,通过净产率直观分析和方差分析获取优化的工艺条件;通过绘制反应速率曲线和寻找化学平衡点来判定反应时间;并采用体外培养方法考察BNO对人鼻咽癌KB、胃癌BGC-803、肝癌SMMC-7721等肿瘤细胞株生长的抑制作用。结果合成最佳反应条件为:马钱子碱(g)与H2O2(mL)配比0.60∶0.75;反应温度50℃;乙醇体积60mL;反应时间220min。在此优化工艺条件下,BNO的产率达112.9%,HPLC面积归一化纯度为98.3%。孵育48h后,BNO对人鼻咽癌KB和胃癌BGC-803细胞增殖有显著的抑制作用,其半抑制浓度(IC50)分别为31.5μmol/L、22.5μmol/L;经形态学分析尚显示BNO对这二株瘤细胞可产生一定的损伤。结论优化工艺条件后一步合成BNO产率高,产物的纯度好,且具有一定的体外抗肿瘤活性。
OBJECTIVE To investigate the optimal conditions of one-step synthesis of brucine N-oxide(BNO) and observe its anti-tumor effects in vitro.METHODS N-oxidation was used to perform one-step synthesis of BNO by taking brucine and hydrogen peroxide(H2O2) as raw materials.Orthogonal experimental design was applied to study the influences of raw material proportion,reaction temperature and solvent(absolute alcohol) volume on the yield of the reaction and purity of target products.The conditions of optimal process were obtained by means of visual and variance analysis of the net yield.The reaction time was given by drawing the chemical reaction rate curve and finding out the equilibrium point.The inhibitory effects of BNO were also investigated in vitro on proliferation of the human cell lines of nasopharyngeal carcinoma KB,stomach carcinoma BGC-803 and liver carcinoma SMMC-7721.RESULTS The optimal synthetic conditions were as follows: the ratio of brucine(g) to H2O2(mL) was 0.60∶0.75;reaction temperature was 50 ℃;alcohol volume was 60 mL;reaction time was 220 min.Under the optimal conditions,the reaction yield reached up to 112.9% and the purity determined by using HPLC area normalization method was 98.3%.After hatching for 48 hours,the proliferation of KB and BGC-803 cells were significantly inhibited by BNO and its hemi-inhibitory concentrations(IC50) were 31.5 μmol/L and 22.5 μmol/L respectively,BNO also caused damage to the two cells through histomorphological analysis.CONCLUSION It is high-yield to synthesize BNO by one-step reaction under the optimal conditions and the products of high purity are prepared.It also has anti-tumor activity in vitro.
出处
《南京中医药大学学报》
CAS
CSCD
北大核心
2011年第5期463-467,共5页
Journal of Nanjing University of Traditional Chinese Medicine
基金
南京中医药大学科技重点培育项目(09XPY03)
江苏省高校"青蓝工程"项目(2008年)