目的:荆防颗粒是新型冠状病毒肺炎预防及轻症治疗的推荐用药,本文结合化学成分分析及活性检测,阐明其潜在的活性成分。方法:采用酶学方法测定荆防颗粒提取物对新型冠状病毒3-chymotrypsin-like protease(3CL^(pro))、papain like protea...目的:荆防颗粒是新型冠状病毒肺炎预防及轻症治疗的推荐用药,本文结合化学成分分析及活性检测,阐明其潜在的活性成分。方法:采用酶学方法测定荆防颗粒提取物对新型冠状病毒3-chymotrypsin-like protease(3CL^(pro))、papain like protease(PL^(pro))、spike protein receptor-binding domain(S-RBD)及人cyclooxygenase-2(COX-2)的抑制活性,利用氨水引咳小鼠模型测试其止咳作用;基于liquid chromatography-mass spectrometry(LC/MS)技术对荆防颗粒进行化学成分定性、定量分析,阐明其化学组成;采用酶学实验、分子对接、定点突变等方法测定荆防颗粒中抑制3CL^(pro)、PL^(pro)的主要活性成分并阐明可能的作用机制。结果:荆防颗粒提取物对新型冠状病毒3CL^(pro)、PL^(pro)蛋白酶具有一定的抑制作用,且具有COX-2抑制活性及止咳药效。进一步在荆防颗粒鉴定了56个成分,其中16个成分经与对照品比对,准确鉴定其化学结构,并测定了其含量,总量为2498.8μg/g。16个成分中的主要成分升麻素苷对3CL^(pro)和PL^(pro)均具有显著的抑制活性,8μmol/L的抑制率分别为76.8%和78.2%;新橙皮苷、柚皮苷对PL^(pro)有抑制活性,8μmol/L的抑制率分别为53.5%和46.1%。分子对接结果表明,升麻素苷可与3CL^(pro)、PL^(pro)活性口袋的氨基酸残基形成氢键,结合能分别为-7.7和-7.3 kcal/mol。定点突变结果表明,氨基酸残基K157是升麻素苷与PL^(pro)相互作用的重要活性位点。结论:荆防颗粒的主要成分升麻素苷、新橙皮苷、柚皮苷等具有抑制新型冠状病毒3CL^(pro)及PL^(pro)的活性,为荆防颗粒的临床合理使用提供了依据。展开更多
Purpose of the Study: COVID-19 is caused by the SARS-CoV-2 virus that had a global pandemic spread in the last two years. Symptoms of the disease include respiratory distress and, in severe cases may consequently lead...Purpose of the Study: COVID-19 is caused by the SARS-CoV-2 virus that had a global pandemic spread in the last two years. Symptoms of the disease include respiratory distress and, in severe cases may consequently lead to death. Blocking the viral proteins can aid in treating this disease and alleviating its symptoms. Two target proteins of the coronavirus that are hot spots in drug discovery are the papain-like protease PL-pro and the main protease M-pro. PL-pro is an enzyme that is required for processing viral polyproteins to generate a functional replicase complex and enable viral spread. M-pro is the major protease of SARS-CoV-2, which is a keystone in viral replication and transcription. Methods: In this study, we shed the light on the route of targeting viral proteins for disease alleviation, by targeting the two aforementioned enzymes, PL-pro and M-pro using in silico studies. Docking experiments, using AutoDock algorithms, were performed to predict the inhibitory effect of recently produced synthetic derivatives of curcumin on the viral proteins. Results: Most of the curcumin derivatives have shown variable levels of inhibition, e.g., S1 - S6, mainly on the papain-like protease, and to a lesser extent on the main protease. Conclusion: The results indicated that curcumin derivatives can be potent anti-viral drug of SARS-CoV-2, namely targeting the papain-like protease.展开更多
To the Editor:The papain-like protease(PL^(pro)),as one of the most important proteases of SARS-CoV-2,has emerged as a highly promising target protein,its inhibitor probably holds dual potentials,namely blocking the c...To the Editor:The papain-like protease(PL^(pro)),as one of the most important proteases of SARS-CoV-2,has emerged as a highly promising target protein,its inhibitor probably holds dual potentials,namely blocking the cleavage of viral polyprotein and intercepting the deubiquitination and deISGylation functions to restore antiviral immunity1.展开更多
文摘目的:荆防颗粒是新型冠状病毒肺炎预防及轻症治疗的推荐用药,本文结合化学成分分析及活性检测,阐明其潜在的活性成分。方法:采用酶学方法测定荆防颗粒提取物对新型冠状病毒3-chymotrypsin-like protease(3CL^(pro))、papain like protease(PL^(pro))、spike protein receptor-binding domain(S-RBD)及人cyclooxygenase-2(COX-2)的抑制活性,利用氨水引咳小鼠模型测试其止咳作用;基于liquid chromatography-mass spectrometry(LC/MS)技术对荆防颗粒进行化学成分定性、定量分析,阐明其化学组成;采用酶学实验、分子对接、定点突变等方法测定荆防颗粒中抑制3CL^(pro)、PL^(pro)的主要活性成分并阐明可能的作用机制。结果:荆防颗粒提取物对新型冠状病毒3CL^(pro)、PL^(pro)蛋白酶具有一定的抑制作用,且具有COX-2抑制活性及止咳药效。进一步在荆防颗粒鉴定了56个成分,其中16个成分经与对照品比对,准确鉴定其化学结构,并测定了其含量,总量为2498.8μg/g。16个成分中的主要成分升麻素苷对3CL^(pro)和PL^(pro)均具有显著的抑制活性,8μmol/L的抑制率分别为76.8%和78.2%;新橙皮苷、柚皮苷对PL^(pro)有抑制活性,8μmol/L的抑制率分别为53.5%和46.1%。分子对接结果表明,升麻素苷可与3CL^(pro)、PL^(pro)活性口袋的氨基酸残基形成氢键,结合能分别为-7.7和-7.3 kcal/mol。定点突变结果表明,氨基酸残基K157是升麻素苷与PL^(pro)相互作用的重要活性位点。结论:荆防颗粒的主要成分升麻素苷、新橙皮苷、柚皮苷等具有抑制新型冠状病毒3CL^(pro)及PL^(pro)的活性,为荆防颗粒的临床合理使用提供了依据。
文摘Purpose of the Study: COVID-19 is caused by the SARS-CoV-2 virus that had a global pandemic spread in the last two years. Symptoms of the disease include respiratory distress and, in severe cases may consequently lead to death. Blocking the viral proteins can aid in treating this disease and alleviating its symptoms. Two target proteins of the coronavirus that are hot spots in drug discovery are the papain-like protease PL-pro and the main protease M-pro. PL-pro is an enzyme that is required for processing viral polyproteins to generate a functional replicase complex and enable viral spread. M-pro is the major protease of SARS-CoV-2, which is a keystone in viral replication and transcription. Methods: In this study, we shed the light on the route of targeting viral proteins for disease alleviation, by targeting the two aforementioned enzymes, PL-pro and M-pro using in silico studies. Docking experiments, using AutoDock algorithms, were performed to predict the inhibitory effect of recently produced synthetic derivatives of curcumin on the viral proteins. Results: Most of the curcumin derivatives have shown variable levels of inhibition, e.g., S1 - S6, mainly on the papain-like protease, and to a lesser extent on the main protease. Conclusion: The results indicated that curcumin derivatives can be potent anti-viral drug of SARS-CoV-2, namely targeting the papain-like protease.
基金National Natural Science Foundation of China(82151219,82130105)the Strategic Priority Research Program of Chinese Academy of Sciences(SIMM010109,SIMM010111)Shanghai Institute of Materia Medica of Chinese Academy of Sciences(SIMM0120231003)for the financial support.
文摘To the Editor:The papain-like protease(PL^(pro)),as one of the most important proteases of SARS-CoV-2,has emerged as a highly promising target protein,its inhibitor probably holds dual potentials,namely blocking the cleavage of viral polyprotein and intercepting the deubiquitination and deISGylation functions to restore antiviral immunity1.
