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目的:评价聚乙二醇化重组人肿瘤坏死因子-α(PEG-rHuTNF-α)的价值。方法:用三种不同分子量的聚乙二醇(PEG)修饰重组人肿瘤坏死因子-α(rHuTNF-α);用凝胶过滤色谱法分离PEG-rHuTNF-α;按L929靶细胞法测定rHuTNF-α和PEG-rHuTNF-α的体外活性;通过S-180实体瘤的坏死研究其体内活性。结果:rHuTNF-α能在温和的条件下用PEG修饰,用分子量为5000的PEG修饰重组人肿瘤坏死因子的主要产物含有四个PEG分子,分子量为12000和20000的PEG修饰的主要产物均含二个PEG分子。用同一种分子量的PEG修饰所得PEG-rHuTNF-α的活性与所含PEG分子的个数有关,所含PEG分子数愈少,则活性愈高;当PEG-rHuTNF-α的分子量相同时,所含单个PEG分子的分子量愈大,活性愈高。当与胰蛋白水解酶混合存放时,rHuTNF-α的活性随时间的延长而降低,但所有PEG-rHuTNF-α对蛋白水解酶有明显的阻抑作用。体内活性测定结果表明:PEG_(5000)-rHuTNF-α(1500 IU/鼠)与rHuTNF-α(3000 IU/鼠)有相近的抗肿瘤活性;PEG_(12000)-rHuTNF-α(1500 IU/鼠)与rHuTNF-α(3000 IU/鼠)比较有较高的诱导肿瘤坏死的效果;PEG_(20000)-rHuTNF-α(1500 IU/鼠)与rHuT-NF-α(6000 IU/鼠)比较有更高的抗肿瘤活性。结论:聚乙二醇化重组人肿瘤坏死因子可能更适合于治疗使用。 (责?
Objective: To evaluate the value of pegylated recombinant human tumor necrosis factor-α (PEG-rHuTNF-α). Methods: Recombinant human tumor necrosis factor-α (rHuTNF-α) was modified with three different molecular weight polyethylene glycol (PEG); PEG-rHuTNF-α was separated by gel filtration chromatography; alpha and PEG-rHuTNF-alpha; their in vivo activity was investigated by necrosis of S-180 solid tumors. Results: rHuTNF-α could be modified with PEG under mild conditions. The main product of recombinant human TNF with PEG of molecular weight 5000 contained four PEG molecules. The main products modified by PEG with molecular weights of 12000 and 20000 contained two PEG molecules. The activity of PEG-rHuTNF-α modified by PEG with the same molecular weight was related to the number of PEG molecules contained. The smaller the number of PEG molecules, the higher the activity. When the molecular weight of PEG-rHuTNF-α was the same, The larger the molecular weight of a single PEG molecule, the higher the activity. When mixed with trypsin, the activity of rHuTNF-α decreased with time, but all PEG-rHuTNF-α had a significant inhibitory effect on proteolytic enzymes. The results of in vivo activity assay showed that PEG_ (5000) -rHuTNF-α (1500 IU / mouse) had similar antitumor activity with rHuTNF-α (3000 IU / mouse) ) Had a higher effect of inducing tumor necrosis compared with rHuTNF-α (3000 IU / mouse); PEG 20000-rHuTNF-α 1500 IU / mouse vs rHuT-NF-α 6000 IU / Higher anti-tumor activity. Conclusion: Pegylated recombinant human tumor necrosis factor may be more suitable for therapeutic use. (responsibility?