inhibitory activity against sars cov 2 3cl protease  (AMS Biotechnology)

Journal: Scientific Reports

Article Title: 9-aminominocycline potentiates the efficacy of EIDD-1931 and PF-332 by targeting the papain like protease enzyme of SARS-CoV-2

doi: 10.1038/s41598-025-89717-3

Figure Lengend Snippet: Inhibition of SARS-CoV-2 PLpro enzyme activities by 9-AMN. The enzymatic inhibitory effects of 9-AMN and minocycline (each at 50 µM) on the SARS-CoV-2 PLpro enzyme were evaluated as detailed in the materials and methods. ( A ) Inhibition of proteolytic activity. ( B ) Inhibition of DUB activity. ( C ) Gradual reduction of proteolytic activity with increasing 9-AMN concentration. ( D ) Gradual decline of DUB activity with increasing 9-AMN concentration. ( E ) Inhibition of 3CLpro enzyme activity ( F ) Inhibition of RdRp enzyme activity. The enzymatic activity of control samples treated with DMSO (0.1%) is taken as 100% for calculating relative enzymatic activity. GRL-0617 (50 µM) treated wells were used as a specificity control. The graph is representative of three independent experiments (n = 3). The P value < 0.05 is denoted as "*" indicating statistical significance, while “ns” refers to statistically not significant.

Article Snippet: The recombinant full-length untagged 3CLpro (cat#100823) and His-tagged PLpro (cat#100735), assay buffers (cat#79956 and cat#78039-1 for 3CLpro and PLpro, respectively), GC-367 (cat#78013), and fluorescently labeled substrates (cat#79952 for 3CLpro, cat#79997 for PLpro protease activity, and cat#82200 for DUB activity) were purchased from BPS Biosciences (San Diego, CA, USA).

Techniques: Inhibition, Activity Assay, Concentration Assay, Control

Journal: Viruses

Article Title: Targeting Viral and Cellular Cysteine Proteases for Treatment of New Variants of SARS-CoV-2

doi: 10.3390/v16030338

Figure Lengend Snippet: In vitro inhibition of 3CL pro activity. Purified recombinant SARS-CoV-2 3CL pro was combined with 500 µM of IBuDM, or IBnDM, BTBnDM, and BFBnDM, and the 3CL pro inhibitor GC376 (2 µM) diluted in assay buffer (50 mM HEPES pH 7.2, 10 mM DTT, and 0.1 mM EDTA) and added to 3CL pro substrate solution (40 µM) in an opaque white 96-well plate for 60 min at 37 °C. DMSO was included as a control and used at 0.5%. Then, reactions were terminated by adding 50 µL of Luciferin Detection Reagent, and after 20 min at room temperature, luminescence was recorded on a GloMax ® luminometer. The results were expressed as relative luminescence units (RLU) ( A ) and percentage of enzymatic activity inhibition ( B ) and represent the mean ± SD based on a minimum of three independent experiments. Statistical analysis utilized GraphPad Prism 8 software (GraphPad Software, San Diego, CA, USA) and involved one-way analysis of variance (ANOVA). Asterisks represent significance levels of the p -values. ** and *** indicate values less than 0.01 and 0.001, respectively.

Article Snippet: Purified recombinant SARS CoV-2 3CL pro (9000 µg/mL) (BPS Bioscience, San Diego, CA, USA, cat# 100823) and purified GST-PL pro (1000 nM) (R&D Systems, Minneapolis, MN, USA, Cat# E-611-050) were used.

Techniques: In Vitro, Inhibition, Activity Assay, Purification, Recombinant, Control, Software

Journal: Viruses

Article Title: Targeting Viral and Cellular Cysteine Proteases for Treatment of New Variants of SARS-CoV-2

doi: 10.3390/v16030338

Figure Lengend Snippet: In vitro inhibition of PL pro activity. Purified recombinant SARS-CoV-2 PL pro was combined with 500 µM of IBuDM, IBnDM, BTBnDM, and BFBnDM, and the 3CL pro inhibitor GC376 (2 µM) and Camostat (5 µM), diluted in assay buffer (50 mM HEPES pH 7.2, 10 mM DTT, and 0.1 mM EDTA) and added to 3CL pro substrate solution (40 µM) in an opaque white 96-well plate for 60 min at 37 °C. DMSO was included as a control and used at 0.5%. Then, reactions were terminated by adding 50 µL of Luciferin Detection Reagent, and after 20 min at room temperature, luminescence was recorded on a GloMax ® luminometer. The results were expressed as relative luminescence units (RLU) ( A ) and percentage of enzymatic activity inhibition ( B ) and represent the mean ± SD based on a minimum of three independent experiments. Statistical analysis utilized GraphPad Prism 8 software (GraphPad Software, San Diego, CA, USA) and involved one-way analysis of variance (ANOVA). Asterisks represent significance levels of the p -values. ** indicates a value less than 0.01.

Article Snippet: Purified recombinant SARS CoV-2 3CL pro (9000 µg/mL) (BPS Bioscience, San Diego, CA, USA, cat# 100823) and purified GST-PL pro (1000 nM) (R&D Systems, Minneapolis, MN, USA, Cat# E-611-050) were used.

Techniques: In Vitro, Inhibition, Activity Assay, Purification, Recombinant, Control, Software

Journal: Marine Drugs

Article Title: Polyphenolic Compounds Isolated from Marine Algae Attenuate the Replication of SARS-CoV-2 in the Host Cell through a Multi-Target Approach of 3CL pro and PL pro

doi: 10.3390/md20120786

Figure Lengend Snippet: Inhibitory activity of isolated compounds on the cell-free cleavage of 3CL pro and PL pro .

Article Snippet: The in vitro inhibition assay kits for 3CL pro and PL pro were purchased from the AMSBIO company (Madrid, Spain).

Techniques: Activity Assay, Isolation

Journal: Marine Drugs

Article Title: Polyphenolic Compounds Isolated from Marine Algae Attenuate the Replication of SARS-CoV-2 in the Host Cell through a Multi-Target Approach of 3CL pro and PL pro

doi: 10.3390/md20120786

Figure Lengend Snippet: Cell-based 3CL pro inhibitory activities of ( a ) Ishophloroglucin A (IPA), ( b ) Dieckol, ( c ) Eckmaxol, and ( d ) Diphlorethohydroxycarmalol (DPHC), and cell-based PL pro inhibitory activities of ( e ) Ishophloroglucin A (IPA), ( f ) Dieckol, ( g ) Eckmaxol, and ( h ) Diphlorethohydroxycarmalol (DPHC). Triplicate experiments were used to evaluate the data and the mean value is expressed with ±SD. * p < 0.05, ** p < 0.01, against PM-treated group or ## p < 0.01, against control (ANOVA, Duncan’s multiple range test).

Article Snippet: The in vitro inhibition assay kits for 3CL pro and PL pro were purchased from the AMSBIO company (Madrid, Spain).

Techniques: