P(HPMA-APMA)-ATRA的合成及其对HL-60细胞诱导分化能力的评估

以N-(2-羟丙基)甲基丙烯酰胺(HPMA),N-(3-氨基丙基)甲基丙烯酰胺(APMA)和全反式维甲酸(ATRA)为原料,采用自由基溶液聚合法设计合成P(HPMA-APMA)-ATRA,并用核磁共振氢谱对该化合物进行结构表征.相比于单体ATRA,聚合物的水溶性显著增加,同时可通过胞吞作用进入细胞.3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法评估聚合物和单体ATRA对人早幼粒白血病细胞HL-60生长的抑制作用,流式细胞术检测两者对HL-60细胞周期分布及细胞表面抗原CD11b表达的影响,进一步结合氯化硝基四氮唑蓝(NBT)还原法评估聚合物诱导HL-60细胞分化的能力.结果显示,聚合物比单体ATRA具有更强的细胞生长抑制活性,其IC50值分别为1.03和4.09μmol/L;聚合物还具有更高的G0/G1期细胞阻滞效应,1.2μmol/L时,聚合物比单体ATRA的G0/G1期细胞率高出17.7%;同样,0.4μmol/L聚合物与2.4μmol/L单体ATRA诱导HL-60的NBT还原能力相当,0.8μmol/L聚合物与2.4μmol/L单体ATRA诱导HL-60细胞表面抗原CD11b表达相当,表明聚合物比单体ATRA具有更强的诱导HL-60细胞向粒细胞分化的能力,其药效增强3~4倍.

Preparation of P（HPMA）-APMA-ATRA and Assessment of Its Induction of HL-60 Cell Differentiation

P（HPMA-APMA）-ATRA was synthesized by free-radical solution polymerization with N-（2-hydroxypropyl methacrylamide） （HPMA）, N-（3-aminopropyl） methacrylamide （APMA） and all-trans retinoic acid （ATRA） as raw materials. The structure of the polymer was confirmed by 1H NMR. Compared with the monomeric ATRA, the water-solubility of the polymer increased significantly and could enter the cells through endocytosis. The inhibition of polymerized and monomeric ATRA on HL-60 cell growth was assessed by MTT（3-（4,5-dimethylthiazol- 2-yl）-2,5-diphenyl-tetrazolium bromide）. Then flow cytometry was used to detect the effects of the two compounds on cell cycle distribution and expression of cell surface antigen CDllb in HL-60 cell. Finally, a further experiment with NBT （nitroblue tetrazolium） was conducted to analyze the cell differentiation of HL-60 induced by the two compounds. The P-ATRA showed a stronger suppressive ability on cell growth than monomeric ATRA, the IC50 was 1.03 and 4.09 μmol L-1, respectively. The P-ATRA had a higher G0/G1 phase cell block effect. At the concentration of 1.2 μmol L-1, G0/G1 phase cell rate induced by P-ATRA was 17.7% higher than that of monomeric ATRA. The NBT reduction ability of HL-60 induced by 0.4 μmol L-1 P-ATRA was similar with that of 2.4 μmol L-： ATRA, the expression of cell surface antigen CD1 lb in HL-60 cell induced by 0.8 μmol L-1 P-ATRA was similar with that of 2.4 μmol L-1 ATRA, and the results indicated that the differentiation of HL-60 cell induced by P-ATRA increased 3-4 folds compared to that of ATRA.