骨靶向药物表阿霉素-氧化葡聚糖-阿仑膦酸钠聚合物的制备及对骨肉瘤细胞的影响  

Preparation and effect of bone targeting drug of the epirubicin-oxidized dextran-alendronate compound for osteosarcoma cells

作　　者：田大为[1] 何宏生[1] 张鹏[1] 黄晓华[1] 赵辉[1] 李刚[1] 熊敏[1] 

机构地区：[1]湖北医药学院附属东风医院骨科,骨科研究所,湖北省十堰442000

出　　处：《中国医师杂志》2014年第11期1498-1502,1507,共6页Journal of Chinese Physician

基　　金：武汉大学研究生资助科研项目（201130302020006）,受中央高校基本科研业务费专项基金资助;湖北省教育厅科学技术研究资助项目（B20112102）

摘　　要：目的 探讨合成骨靶向药物表阿霉素-氧化葡聚糖-阿仑膦酸钠聚合物对骨肉瘤细胞的影响.方法 （1）根据席弗碱原理合成聚合物.（2）红外、紫外和核磁对聚合物进行表征,测量氧化葡聚糖的分子量,醛基含量,阿仑膦酸钠-氧化葡聚糖聚合物中阿仑膦酸的含量,表阿霉素在聚合体中的载药量,体外羟基磷灰石结合实验验证亲骨性.（3）四甲基偶氮唑蓝（MTT）实验验证聚合物对骨肉瘤细胞的细胞毒性,流式细胞仪探讨聚合物对凋亡作用的影响.结果 物质各有特殊的物理表征.氧化葡聚糖重均分子量（MW）为4 251 ±68,数均分子量（MN）为2 551 ±42,分子量分布宽度为1.78.氧化葡聚糖的醛基含量为（10.41±0.2）mmol/g.合成药物中表阿霉素（EPI）的载药量为（5.28±0.23）％.醛基与阿仑膦酸钠（ALN）反应的最佳物质量比（投料比）应在5：2～3之间.ALN-Dex-EPI和EPI的羟基磷灰石（HA）吸附率分别为86.13％,13.91％.MTT实验中,ALN-Dex-EPI、EPI、ALN的IC50（单位：μg/ml）在24、48、72 h分别为0.142,0.505,0.219;0.251,0.739,0.518;45.21,27.97,18.96.在流式细胞实验中,EPI、ALN-Dex-EPI、ALN、Dex凋亡及坏死百分率（％）为80.13±4.05,97.01±2.58,31.53±6.9,14.01±2.51,组间比较差异有统计学意义.结论 成功合成的骨靶向药物具有良好的体外亲骨性外,对骨肉瘤细胞有较强的细胞毒性和促进凋亡作用.Objective To prepare a new bone-targeted drug of epirubicin-oxidized dextran-alendronate compound,and explore the effect of compound on osteosarcoma cells.Methods （1） Based on Schiff＇s base theory,we synthesized the compound.（2） We investigated physicochemical character with ultraviolet （UV）,infrared （IR）,and 1H-nuclear magnetic resonance （1H-NMR）,molecular weight and aldehyde group content in oxidized dextran,alendronate content in oxidized dextran-alendronate compound,epirubicin drug loading capacity in the compound,and rate of charge for three matters,and affinity to bone in vitro though Hydroxyapatite （HA） absorbing test.（3） We investigated the compound＆#39;s cytotoxicity effect through methyl thiazolyl tetrazolium （MTT） test,apoptosis influence through flow cytometry （FCM）.Results （1） There were typical physicochemical characters.（2） Oxidized dextran molecular weight （MW） was 4 251 ± 68,number of average molecular weight （MN） was 2 551 ± 42,and molecular weight distribution width was 1.78.Aldehyde group content of oxidized dextran was （10.41 ± 0.2）mmol/mg,epirubicin drug loading capacity was （5.28 ± 0.23） %,rate of charge was 5,and 2 ～ 3 between aldehyde group content in oxidized dextran and alendronate （ALN）.（3） In the MTT test,epirubicin （EPI）-Dex-ALN,EPI,ALN obviously restrained osteosarcoma cell proliferation,the minimum concentration（μg/ml） was 1,1,and 40; IC50 （μg/ml） of EPI-Dex-ALN,EPI,ALN was 0.142,0.505,0.219; 0.251,0.739,and 0.518; 45.21,27.97,and 18.96 after 24 h,48 h,72 h; in FCM （flow cytometry）,apoptosis and necrosis rate of EPI-Dex-ALN,ALN,and Dex was 80.13 ±4.05,97.01 ±2.58,31.53 ±6.9,and 14.01 ±2.51.Conclusions We successfully synthesized a new bone-targeted drug delivery system,which showed better bone affinity in vivo,and kept biological activity with powerful targeted function.

关 键 词：表柔比星/药理学 葡聚糖类/药理学 二膦酸盐类/药理学 聚合物 骨肉瘤/病理学 

分 类 号：R738.1[医药卫生—肿瘤]