摘要
目的:制备靶向肿瘤细胞促性腺激素释放激素(GnRH)受体的多肽-药物偶联物。方法:将GnRH的6位甘氨酸(Gly~6)用半胱氨酸(Cys)替代,10位的甘氨酰胺(Gly~(10)-NH_2)分别用乙胺和氨基脲取代,合成了2个GnRH类似物。以硫醚键将上述GnRH类似物与阿霉素(Dox)偶联,得到了2个多肽-药物偶联物[Cys~6-desGly~(10)-Pro~9-NHEt]-GnRH-Dox(偶联物1)和[Cys~6-α-azaGly~(10)-NH_2]-GnRH-Dox(偶联物2)。MTT实验考察2个偶联物对GnRH受体过表达的MCF-7人乳腺癌细胞的体外增殖抑制活性,GnRH受体抑制实验评价偶联物2的靶向性。结果:2个偶联物对MCF-7细胞的增殖抑制活性均低于阿霉素,偶联物2对MCF-7细胞的增殖抑制活性高于偶联物1。结论:偶联物2的抗肿瘤活性可能是由[Cys~6-α-azaGly~(10)-NH_2]-GnRH与GnRH受体的靶向结合而介导的细胞内吞摄取作用而实现。
Objective:To prepare peptide-drug conjugates targeting gonadotropin-releasing hormone(GnRH)receptor on tumor cells.Methods:Two new GnRH analogs were synthesized by replacing the Gly~6 with Cys and substitution of C-terminal Gly~(10)-NH_2 by ethylamide andα-azaGly~(10)-NH_2,respectively.The GnRH analogs were conjugated with doxorubicin(Dox)through a linker containing thioether bond to afford two GnRH-Dox conjugates,named[Cys~6-desGly~(10)-Pro~9-NHEt]-GnRH-Dox(conjugate 1)and[Cys~6-α-azaGly~(10)-NH_2]-GnRH-Dox(conjugate2),respectively.The direct in vitro growth inhibitory effect of these two conjugates on MCF-7 human breast cancer cells which overexpress GnRH recepter was examined by MTT assay,and the targeting effect of conjugate 2 was evaluated by GnRH receptor inhibition test.Results:Both of the two conjugates exhibited lower antiproliferative activity against MCF-7 cell line in comparison with Dox,and the antiproliferative activity of conjugate 2 was higher than that of conjugate 1.Conclusion:The antiproliferative effect of conjugate 2 on MCF-7 cells might be due to the cellular uptake mediated by the target binding of[Cys~6-α-azaGly~(10)-NH_2]-GnRH to GnRH receptors.
引文
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