新型ADEPT前药头孢美法仑的合成及其体外抗肿瘤活性评价

发布时间：2018-05-10 06:06

本文选题：ADEPT + 前药　；参考：《北京协和医学院》2014年硕士论文

【摘要】：抗体导向的酶前体药物疗法(ADEPT)利用抗体-抗原、酶-前药特异性来介导细胞毒性药物对肿瘤部位的选择性杀伤作用。其中,酶具有低免疫原性及高效性,前药低毒并可被相应的酶高效转化,对肿瘤细胞的杀伤作用呈浓度依赖型。因此,本实验根据低免疫原性的突变性β-内酰胺酶的底物专一性,将β-内酰胺酶的良好底物头孢菌素类与氮芥类细胞毒药物美法仑拼合,设计合成目标化合物头孢美法仑,并对其体外抗肿瘤活性进行初步评价。我们选择GCLE作为原料,将其经碘代反应制成活性较高的GIE,而后与已酯化的美法仑进行偶联,再将偶联物进行氧化和水解从而合成头孢美法仑,终产物采用1H NMR及质谱确证其结构。体外抗肿瘤活性实验采用MTT毒性法,在设定浓度范围内测定前药和原药的抑制率(%)和IC50(μmol/l),结果表明：单前药与空白对照组的结果不存在显著性差异,在体外基本无毒；酶解前药和阳性对照药均对该肿瘤细胞具有显著性杀伤作用,IC50分别为101.97±1.70μ mol/l和66.69±0.37μmol/l;酶解前药对肿瘤细胞的杀伤作用呈浓度依赖性。由此可见,本实验设计合成的头孢美法仑前药与原药美法仑相比,细胞毒作用相差不多,但由于ADEPT系统的靶向性,具有体外无毒及靶向杀伤等优点,是一颇具前景的肿瘤治疗候选药物。
[Abstract]:Antibody-directed enzyme precursor drug therapy (ADEPTT) mediates the selective killing effect of cytotoxic drugs on tumor sites by using antibody-antigen and enzyme-prodrug specificity. Among them, the enzyme has low immunogenicity and high efficiency. Prodrug has low toxicity and can be transformed by the corresponding enzyme. The killing effect of prodrug on tumor cells is concentration-dependent. Therefore, according to the substrate specificity of low immunogenicity mutant 尾 -lactamases, cephalosporins, a good substrate of 尾 -lactamases, were combined with melfarin, a cytotoxic drug of nitrogen mustard, to design and synthesize the target compound cefmefacalam. The anti-tumor activity in vitro was evaluated. We selected GCLE as the raw material, and then synthesized cefmefacalen by oxidation and hydrolysis of the coupling material, which was prepared by iodine substitution reaction, and then was coupled with the esterified mefacalen. The final product was characterized by 1H NMR and mass spectrometry. In vitro antitumor activity test was carried out by MTT toxicity method. The inhibitory rates of prodrug and prodrug were determined within a given concentration range) and IC50 (渭 mol / L). The results showed that there was no significant difference between the single prodrug and the blank control group, and the results were basically non-toxic in vitro. The IC50 of pre-enzymatic drugs and positive control drugs were 101.97 卤1.70 渭 mol/l and 66.69 卤0.37 渭 mol / L, respectively. It can be seen that the cytotoxicity of cefmefacalen prodrug designed and synthesized in this experiment is not much different from that of the original drug, but because of the targeting of ADEPT system, it has the advantages of non-toxic and targeted killing in vitro, and so on. It is a promising candidate for cancer therapy.
【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R914.5;R96

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