蒜氨酸微丸-蒜酶肠溶双层片制备工艺优选及体外抗肿瘤作用评价

Optimization of Preparation Process of Enteric-Coated Alliin Pellets-Alliinse Bilayer Tablets and in Vitro Evaluation of Their Anti-Tumor Effect

目的优选蒜氨酸微丸-蒜酶肠溶双层片的制备工艺,并评价其体外抗肿瘤作用。方法以蒜氨酸的累积释放率为评价指标,以崩解剂种类和用量、肠溶衣用量及微晶纤维素(MCC)与蒜氨酸的质量比为考察因素,采用单因素试验优选蒜氨酸微丸的制备工艺;以大蒜辣素产率为评价指标,以蒜酶层中MCC的用量、包衣温度为考察因素,采用单因素试验优选蒜氨酸微丸-蒜酶肠溶双层片的制备工艺;采用噻唑蓝比色法评价不同质量浓度蒜氨酸微丸-蒜酶肠溶双层片对肿瘤细胞L02,Bel-7402,HL-60,HeLa的生长抑制作用,并计算其细胞生长抑制率。结果蒜氨酸微丸的最优制备工艺,以5%羧甲基淀粉钠为崩解剂,以2%十二烷基硫酸钠为润湿剂,MCC与蒜氨酸质量比为1∶1(m/m);蒜氨酸微丸-蒜酶肠溶双层片的最优制备工艺为,MCC的用量为每片72 mg,包衣温度为25℃,包衣技术为挤出滚圆法。21.98~131.89μg/mL(以大蒜辣素计)蒜氨酸微丸-蒜酶肠溶双层片对L02细胞的生长抑制率不超过10%,以L02组为对照,其对Bel-7402,HL-60,HeLa肿瘤细胞的生长抑制率均显著升高(P <0.05),且呈浓度依赖关系。结论优选的蒜氨酸微丸-蒜酶肠溶双层片的制备工艺稳定、可行,制备的产品具有明显体外抗肿瘤作用。

Objective To optimize the preparation process of Enteric-Coated Alliin Pellets-Alliinse Bilayer Tablets,and to evaluate their anti-tumor effect in vitro.Methods The preparation process of alliin pellets was optimized by the single factor test with the cumulative release rate of alliin as the evaluation index,and with the type and dosage of disintegrating agent,the entericcoating dosage and the mass ratio of microcrystalline cellulose(MCC) to alliin as the investigation factors.The preparation process of Enteric-Coated Alliin Pellets-Alliinse Bilayer Tablets was optimized by the single factor test with the yield of allicin as the evaluation index,and with the amount of MCC in alliinase layer and coating temperature as the investigation factors.The inhibitory effect of Enteric-Coated Alliin Pellets-Alliinse Bilayer Tablets with different mass concentrations on the growth of tumor cells L02,Bel-7402,HL-60 and HeLa was evaluated by the methyl thiazolyl tetrazolium(MTT) assay,and the cell growth inhibition rate was calculated.Results The optimal preparation process of alliin pellets was as follows:5% sodium carboxymethyl starch was taken as the disintegrating agent and 2% sodium dodecyl sulfate was taken as the wetting agent,and the mass ratio of MCC to alliin was 1-1(m/m).The optimal preparation process of Enteric-Coated Alliin Pellets-Alliinse Bilayer Tablets was as follows:the dosage of MCC was 72 mg per tablet,the coating temperature was 25℃,and the coating technology was extrusion spheronization method.The growth-inhibitory rate of 21.98-131.89 μg/mL(calculated by allicin) of Enteric-Coated Alliin Pellets-Alliinse Bilayer Tablets on L02 cells was not more than 10%.Compared with those in the L02 group,the growthinhibitory rates on Bel-7402,HL-60 and HeLa tumor cells increased significantly(P <0.05),which showed a concentrationdependent manner.Conclusion The optimal preparation process of Enteric-Coated Alliin Pellets-Alliinse Bilayer Tablets is stable and feasible,and the prepared product has an obvious anti-tumor effect in vitro.