【作者】 陈鹏；

【导师】 陈河如；

【作者基本信息】 暨南大学 ， 药物化学， 2013， 硕士

【摘要】 目的:以成纤维激活蛋白α（FAP-α）为靶标，化学合成系列靶向FAP-α系列酶激活式抗肿瘤前体寡肽。一方面希望改造后的抗肿瘤寡肽有更强的抗肿瘤活性和一定的靶向性，另一方面希望改造后的抗肿瘤寡肽能克服多肽类药物在体内易酶解的确定缺点，延长其在血清中的半衰期，使得改造后的抗肿瘤多肽更加具有成药性。方法：1.采用Fmoc/t-Bu固相多肽合成法合成YSL、YSV、LDV、YIGSR母体寡肽，通过羧端酰胺化、N端封闭特殊靶向二肽片段Z-GP两种方法对母体寡肽进行结构改造。用高效液相进行分离纯化。2.利用RP-HPLC监控样品在37℃、不同时间点的浓度变化，以此考察母体肽和结构修饰寡肽在PBS、DMEM和血清中的稳定性。3.通过MTT法检测母体肽及改造的抗肿瘤寡肽的肿瘤细胞增殖抑制活性,并考察Z-GP片段是否封闭了母体肽的活性位点。结果：成功合成YSL、YSV、LDV、YIGSR四条抗肿瘤母体肽、四条C端酰胺化改造的抗肿瘤寡肽和四条N端经Z-GP封闭改造的抗肿瘤寡肽。应用现代波谱学方法（1H-NMR、13C-NMR、ESI-MS等）对所合成的寡肽化合物进行结构确认。大鼠血清稳定性实验表明经过改造后的抗肿瘤短肽在血清中的稳定性明显高于母体肽。MTT实验表明经过结构改造的YSL具有比母体肽强的细胞毒性。更多还原

【Abstract】 Objects:To design and synthesize a series of antitumor pro-peptides，which target on fibroblastactivated protein α. On one hand, it is expected that the modified antitumor pro-peptides willshow enhanced targeting ability in the purpose of increasing their therapy index; while on theother hand, the stability of all the designed polypeptides in serum will be improved，resulted inlonger half-life in serum. To make the modified antitumor polypeptides more drug-like.Methods:1. To synthesize YSL、YSV、LDV and YIGSR parent peptides by solid phase peptide synthesis(spps) based on Fmoc/t-Bu strategy. The parent peptides were modified in two ways: one isto change C-terminal carboxyl into amide; the other is to block N-terminal of the parentpeptides by a fragment, namely Z-GP, which is recognized specifically by FAP-. All theprepared peptide were purified by RP-HPLC.2. The stability of both parent peptides and modified antitumor oligopeptides were evaluated bymonitoring their changing concentration at scheduled time points using RP-HPLC method, inwhich each sample was dissolved in PBS、DMEM and serum and bathed at the temperatureof37℃respectively.3. Cytotoxicity of both parent peptides and modified antitumor oligopeptides were determinedby MTT assay respectively to evaluate their antitumor activity, and to evaluate whether ifZ-GP fragment blocks the anti-tumor activity of the parent peptides.Results:Successfully synthesized12oligopeptide. The Structure were Confirm by1H-NMR、13C-NMRand ESI-MS. Stability test show that modified antitumor oligopeptides were more stable thanParent peptides. MTT test indicated that modified YSL had stronger cytotoxicity than parentpeptide.更多还原