白蛋白美登素偶联物的制备及其体外抗胰腺癌活性研究  

Preparation and in vitro anti-pancreatic cancer activity study of albumin-maytansine conjugate

作　　者：胡尚玖 王莹[1] 张国宁[1] 朱梅[1] 王明华[1] 王菊仙[1] HU Shang-jiu;WANG Ying;ZHANG Guo-ning;ZHU Mei;WANG Ming-hua;WANG Ju-xian(ic Synthesis Chamber,Institute of Medicinal Biotechnology,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100050,China)

机构地区：[1]中国医学科学院北京协和医学院医药生物技,术研究所有机合成室,北京100050

出　　处：《中国医药生物技术》2022年第5期393-404,共12页Chinese Medicinal Biotechnology

基　　金：中国医学科学院医学与健康科技创新工程(2021-I2M-1-026)。

摘　　要：目的制备人血清白蛋白(HSA)与美登素衍生物mertansine(DM1)的偶联物,评价其体外抗胰腺癌细胞活性。方法以安丝菌素为原料经还原、酯化等反应制得美登素衍生物DM1,以N-羟基琥珀酰亚胺与马来酸酐为原料反应制得连接子SMCC,通过连接子SMCC将DM1与人血清白蛋白偶联制得偶联物HSA-DM1,采用HPLC、MALDI-TOF对其进行表征并计算平均偶联比。采用CCK-8法测定偶联物对MIA PaCa-2、PANC-1和BxPC-3三种胰腺癌细胞系的增殖抑制作用;利用流式细胞术和Westernblot法测定其对MIAPaCa-2和PANC-1细胞的促凋亡能力。结果成功制备偶联物HSA-DM1,其平均偶联比为3.32,对MIAPaCa-2、PANC-1和Bx PC-3三种细胞系的IC50值分别为(48.2±6.5)、(55.0±4.7)、(84.0±11.8)nmol/L,同时其对MIA PaCa-2和PANC-1细胞的促凋亡作用呈剂量依赖。结论偶联物HSA-DM1具有显著抑制胰腺癌细胞增殖的活性,具有潜在的临床应用前景。Objective To prepare human serum albumin(HSA)-mertansine derivative(DM1)conjugate and evaluate its anti-pancreatic cancer cells activity in vitro.Methods Maytansine derivative DM1 was synthesized by reduction and esterification reaction from ansamitocin.The linker SMCC was synthesized by reaction of N-hydroxysuccinimide and maleic anhydride.HSA-DM1 was prepared by coupling DM1 with human serum albumin through the linker SMCC.HPLC and MALDI-TOF were used to characterize and calculate the average coupling ratio.CCK-8 method was used to detect the inhibitory effect of the conjugate on the proliferation of three pancreatic cancer cell lines:MIA PaCa-2,PANC-1 and BxPC-3.Flow cytometry and Western blot were used to detect its proapoptotic effect on MIA PaCa-2 and PANC-1 cells.Results Successful preparation of HSA-DM1 conjugate and it’s average coupling ratio was 3.32.The IC50 values of HSA-DM1were(48.2±6.5),(55.0±4.7)and(84.0±11.8)nmol/L for MIA PaCa-2,PANC-1 and BxPC-3 cell lines,respectively.And the proapoptotic effect on MIA PaCa-2 and PANC-1 cells was dose-dependent.Conclusion The conjugated compound HSA-DM1 has significant inhibitory activity on the proliferation of pancreatic cancer cells and has potential clinical application prospect.

关 键 词：人血清白蛋白 美登素衍生物 偶联物 胰腺癌 

分 类 号：R943[医药卫生—药剂学]