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
SARS CoV 3CL^pro is known to be a promising target for development of therapeutic agents against the severe acute respiratory syndrome （SARS）. A quinolinone compound 1 was selected via virtual screening, and it was ...SARS CoV 3CL^pro is known to be a promising target for development of therapeutic agents against the severe acute respiratory syndrome （SARS）. A quinolinone compound 1 was selected via virtual screening, and it was syn- thetized and tested for enzymatic inhibition in vitro. Compound 1 showed potent inhibitory activity （ICs0= 0.44 μmol/L） toward SARS CoV 3CL^pro. Further work on a series of quinolinone derivatives resulted in the discovery of the most potent compound 23, inhibiting SARS CoV 3CL^pro with an IC50 of 36.86 μmol/L. The structure-activity relationships were also discussed.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SARS CoV 3CL^pro is known to be a promising target for development of therapeutic agents against the severe acute respiratory syndrome （SARS）. A quinolinone compound 1 was selected via virtual screening, and it was syn- thetized and tested for enzymatic inhibition in vitro. Compound 1 showed potent inhibitory activity （ICs0= 0.44 μmol/L） toward SARS CoV 3CL^pro. Further work on a series of quinolinone derivatives resulted in the discovery of the most potent compound 23, inhibiting SARS CoV 3CL^pro with an IC50 of 36.86 μmol/L. The structure-activity relationships were also discussed.
