摘要
目的考察合成的含靶肽(P-FU-PEPTIDE)和不含靶肽(P-FU)的水溶性HPMA聚合物-5-FU的体外释药规律及抗肿瘤活性。方法以不同pH缓冲液为介质,考察两种化合物的稳定性;以小鼠血浆为介质,考察两种化合物中5-FU的释放规律;以人头颈鳞癌和乳腺癌细胞株为模型,考察化合物对不同肿瘤细胞的生长抑制作用;通过TUNEL试验,初步研究原药和化合物的体外抗肿瘤机制。结果化合物在偏酸性环境中均较稳定,稳定性随pH增大而降低,在偏碱性环境中易释药。通过拟合多种释药数据模型,P-FU、P-FU-PEPTIDE在血浆中的释药过程均符合Higuchi方程。在高浓度点时,化合物对4种细胞株均有较好的生长抑制效果。除CNE1外,P-FU、P-FU-PEPTIDE的IC50均较5-FU小(P<0.05),尤其对于SCC9肿瘤细胞,P-FU-PEPTIDE具有较强的选择抑制作用,IC50是P-FU的1/1.37,并具时间依赖性和浓度依赖性。药物对SCC9肿瘤细胞作用24 h后,P-FU-PEPTIDE、P-FU、5-FU组的细胞凋亡率分别为43.10%±2.41%、15.40%±1.13%、2.00%±1.58%。结论 HPMA聚合物-5-FU能增强5-FU对SCC9、Hep2和MCF-7的抑制作用,P-FU-PEPTIDE对SCC9肿瘤细胞比P-FU具更强的选择抑制作用,可能引起该类肿瘤细胞发生凋亡。
OBJECTIVE To investigate in vitro release behaviors, in vitro cytotoxicity and therapeutic efficacy of HPMA copolymer （peptide） - 5 - fluorouracil conjugates （ P - FU - PEPTIDE） and HPMA copolymer - 5 - FU conjugates without targeting moiety （ P - FU）. METHODS The medium of different phosphate buffers were used to determine the in vitro stability of P - FU and P - FU - PEPTIDE. The in vitro release behavior was evaluated by determining the amount of 5 - FU released from P - FU and P - FU - PEP- TIDE in mice plasma at 37℃. Cytotoxicity was evaluated by using a serial cell line of human head and neck squamous cell carcinomas and human breast cancer. The apoptosis of P - FU and P - FU - PEPTIDE was investigated by TUNEL method. RESULTS HPMA copolymer - 5 - FU conjugates were more stable in acidic phosphate buffers. While in slightly basic environment, most of the drugs were released. The release data of P - FU and P - FU - PEPTIDE in plasma could be fitted to Higuchi equation. At high concentration of the in vitro cytotoxicity, P - FU and P - FU - PEPTIDE could increase the cytotoxicity of 5 - FU against all the cells. Except of CNE1, the ratios of IC50 of P- FU -PEPTIDE and P- FU were smaller than the 5 -FU＇s （P 〈 0.05 ）. Sepecially for SCC9, P- FU -PEPTIDE could selectively inhibit apoptosis,the IC50 value is 1/1.37 of P - FU, and the results demonstrated that the cytotoxcity of P - FU - PEPTIDE was concentration - dependent and time - dependent. The treatment with drug in SCC9 for 24 h, P - FU - PEPTIDE displayed significantly stronger inhibition of the cell growth（43.10%±2.41% ） than that of 5 - FU（2.00% ± 1.58% ） group and P - FU group （ 15.40%± 1.13% ）. CONCLUSION It is possible to deliver anticancer drug to the tumor tissue by HPMA copolymer as carrier. Meanwhile, the mechanism of selective inhibition of SCC9 of P - FU - PEPTIDE may due to induced apoptosis.
出处
《华西药学杂志》
CAS
CSCD
北大核心
2014年第3期229-233,共5页
West China Journal of Pharmaceutical Sciences
基金
国家自然科学基金资助项目(批准号:81072600)
