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胰蛋白酶体外筛选天然化合物抗肿瘤活性

何佳丽, 张金秋, 王思懿, 张铮, 张德兴, 钱俊青^*

浙江工业大学药学院，杭州 310032

摘要

为考察胰蛋白酶体外筛选天然化合物抗肿瘤活性实验的可行性，本研究采用猪胰蛋白酶进行了以酶活性抑制为标志的天然化合物筛选试验。选择大豆苷、染料木苷、苦参碱与苦参素4种化合物为试验对象，以抗肿瘤天然药物喜树碱为对照，采用直接检测胰蛋白酶活力抑制及考察活力抑制的胰蛋白酶水解牛血清白蛋白两种实验方法检验抑制效果。实验结果可较好显示天然化合物对胰蛋白酶的抑制效果，且4种天然化合物的抑制率有明显差别。胰蛋白酶活力抑制的检测结果表明苦参碱的抑制效果优于苦参素，显示了胰蛋白酶有较好的活性筛选分辨率，且抑制率随化合物与胰蛋白酶的比例增大有较明显的提高，体现出筛选方法的构效相关性与量效相关性，表明胰蛋白酶作为标靶的抗肿瘤活性筛选实验方法具有良好的应用价值。猪胰蛋白酶与人胰蛋白酶分子结构及性能相似，用于天然化合物抗肿瘤药效筛选易推广，对天然药效化合物的筛选和利用有实际意义。

关键词： 胰蛋白酶; 药效化合物; 体外筛选; 喜树碱; 抗肿瘤

In vitro screening of natural compounds with antitumor activity by trypsin

HE Jia-Li, ZHANG Jin-Qiu, WANG Si-Yi, ZHANG Zheng, ZHANG De-Xing, QIAN Jun-Qing^*

School of Pharmacy, Zhejiang University of Technology, Hangzhou 310032, China

Abstract

In order to investigate the feasibility of in vitro screening the antitumor activity of natural compounds by trypsin, porcine trypsin was used to for screening test, which is marked by inhibition of enzyme activity. Four compounds, namely daidzin, genistin, matrine and oxymatrine, were selected as test subjects. The natural antitumor drug camptothecin was used as the control. The inhibitory effect was detected by two experimental methods: direct detection of trypsin activity inhibition and hydrolysis of bovine serum albumin by trypsin. The results showed the inhibitory effects of the four natural compounds on trypsin, and the inhibition rates of the four natural compounds were significantly different. The enzyme activity assay showed that the inhibitory effect of matrine was better than that of oxymatrine, indicating that trypsin had a good screening resolution. The inhibitory effect was significantly increased with the increased ratio of sample to trypsin, suggesting the structure-activity correlation and dose-effect correlation of the screening methods. Altogether, the experimental method of screening antitumor activity of natural compounds by trypsin has good application values. Since porcine trypsin is similar to human trypsin in terms of molecular structure and performance, it is more applicable for screening of antitumor efficacy of natural pharmacodynamic compounds.

Key words: Trypsin; pharmacological compounds; in vitro screening; camptothecin; antitumor

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通讯作者：钱俊青　　E-mail: qjq@zjut.edu.cn; 2112007172@zjut.edu.cn

引用本文：

何佳丽, 张金秋, 王思懿, 张铮, 张德兴, 钱俊青. 胰蛋白酶体外筛选天然化合物抗肿瘤活性[J]. 生理学报 2022; 74 (6): 1014-1022.

HE Jia-Li, ZHANG Jin-Qiu, WANG Si-Yi, ZHANG Zheng, ZHANG De-Xing, QIAN Jun-Qing. In vitro screening of natural compounds with antitumor activity by trypsin. Acta Physiol Sin 2022; 74 (6): 1014-1022 (in Chinese with English abstract).