The papain-like protease (PLpro) can process the viral polyprotein in a coordinated manner and strip ubiquitin and ISG15 from host-cell proteins to aid coronaviruses in their evasion of the host innate immune responses. Therefore, PLpro is an attractive antiviral drug target for its essential role for coronaviral replication[2]. PLpro inhibitor is a potent inhibitor against the PLpro from the coronavirus that causes severe acute respiratory syndrome (SARS-CoV). PLpro inhibitor was found to have an IC50 value of 2.6 ± 0.1 μM. PLpro inhibitor displayed significant antiviral activity with an EC50 value of 13.1 ± 0.7 uM, without toxicity up to the highest concentration tested. Notably, the increasing antiviral potency correlated with the in vitro inhibition of PLpro, suggesting that the compound works directly on the enzyme in cellshttp://www.google.com/patents/WO2010022355A1cl=en.
PLpro inhibitor 细胞实验
Cell Line
Concentration
Treated Time
Description
References
Vero cells
11 µM and 33 µM
72 hours
Assess the antiviral activity of PLpro inhibitors
EMBO J. 2020 Sep 15;39(18):e106275.
Vero E6 cells
68.2 µM
Evaluate the antiviral effect of GRL0617 (1) in Vero E6 cells
Sci Adv. 2024 Aug 30;10(35):eado4288.
Vero cells
10 µM
24 hours
Evaluate the inhibitory effect of F0213 on SARS-CoV-2 NP antigen, results showed significant inhibition of viral replication.
Protein Cell. 2022 Dec;13(12):940-953.
HEK293T cells
20-80 µM
24 hours
To assess whether GRL0617 can inhibit the in cyto deubiquitinating and deISGylating activity of SARS-CoV-2 PLpro. Results showed that GRL0617 partially recovered poly-ubiquitin-conjugates and ISG15-conjugates.
Nat Commun. 2021 Jan 20;12(1):488.
Vero E6 cells
0.17 to 3.18 µM
24 hours
Evaluate antiviral activity, all four compounds showed strong antiviral effects
Protein Cell. 2021 Nov;12(11):877-888.
Vero E6 cells
25, 75, 100, 200, 300, 400 µM
24 hours
To evaluate the antiviral potential of ATA, IC50 was 50 μM
Int J Biol Macromol. 2023 Mar 1;230:123347.
HEK293T cells
10 µM
24 hours
Evaluate the inhibitory effect of PLpro inhibitors on nsp1-nsp2 protein cleavage
Cell Chem Biol. 2021 Jun 17;28(6):855-865.e9.
Differentiated normal human bronchial epithelial (dNHBE) cells
13.9 nM
3 days
Evaluate the antiviral effect of PF-07957472 (4) in dNHBE cells
Sci Adv. 2024 Aug 30;10(35):eado4288.
Vero
0.41 µM
4 days
Determine antiviral EC50 of WEHI-P4 by plaque assay, showing EC50 of 410 nM
Nat Commun. 2025 Apr 3;16(1):2900.
Vero E6 cells
100 µM
48 hours
To evaluate the antiviral activity of GRL0617 against SARS-CoV-2. Results showed that 100 μM GRL0617 inhibited over 50% of viral replication.
Nat Commun. 2021 Jan 20;12(1):488.
Vero E6 cells
1.1 µM
48 hours
Assessed the protective effect of compound 7 against SARS-CoV-2-induced cell death, with an EC50 of 1.1 μM, comparable to remdesivir
Nat Commun. 2023 Mar 28;14(1):1733.
A549-hACE2 cells
2.5 µM (EC50)
48 hours
Evaluate the antiviral activity of PLpro inhibitors in A549-hACE2 cells, results showed compound 10 exhibited broad-spectrum antiviral effects against SARS-CoV-2 original strain and variants
J Med Chem. 2024 Aug 22;67(16):13681-13702.
HACE2-HeLa cells
5 µM
48 hours
Evaluate the inhibitory effect of PLpro inhibitors on SARS-CoV-2 replication
Cell Chem Biol. 2021 Jun 17;28(6):855-865.e9.
VeroE6-TMPRSS2 cells
2.2–4.8 µM
72 hours
Evaluate the inhibitory effect of F0213 on SARS-CoV-2 variants, results showed dose-dependent inhibition against all variants.
Protein Cell. 2022 Dec;13(12):940-953.
LT-A549
8.352 µM
72 hours
Evaluation of cytotoxicity of compound 5 on LT-A549 cells, showing CC50 of 45.775 μM.
Eur J Med Chem. 2023 Apr 5;252:115272.
Wi-38
10.356 µM
72 hours
Evaluation of cytotoxicity of compound 5 on Wi-38 cells, showing CC50 of 51.781 μM.
Eur J Med Chem. 2023 Apr 5;252:115272.
Caco-2 cells
0.26 µM
Evaluated the antiviral activity of compound 7 in human cells, with an EC90 of 0.26 μM against the USA-WA1/2020 strain
Nat Commun. 2023 Mar 28;14(1):1733.
PLpro inhibitor 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
C57BL/6 mice
SARS-CoV-2 MA10 infection model
Intraperitoneal injection
10 mg/kg
Twice daily for 2 days
Evaluate the antiviral efficacy of PLpro inhibitors in Mice model, results showed compound 10 significantly reduced lung viral load
J Med Chem. 2024 Aug 22;67(16):13681-13702.
C57BL/6 mice
SARS-CoV-2 P21 infection severe disease model
Oral
100 mg/kg and 150 mg/kg
Administered at 6, 24, and 48 hours post-infection
Evaluate efficacy of WEHI-P8 in severe SARS-CoV-2 infection model, results showed both 100 mg/kg and 150 mg/kg doses significantly reduced viral load and alleviated lung inflammation
Nat Commun. 2025 Apr 3;16(1):2900.
Syrian hamsters
SARS-CoV-2 infection model
Oral
15, 30, 45 mg/kg
Once daily for 4 days
To evaluate the in vivo antiviral potential of ATA, results showed reduction in viral load in throat swabs but no significant changes in lung tissues
Int J Biol Macromol. 2023 Mar 1;230:123347.
Mice
BALB/c mice
Oral
20, 50, 150 mg/kg
Twice daily for 4 days
Evaluate the antiviral effect of PF-07957472 in a Mice infection model
Sci Adv. 2024 Aug 30;10(35):eado4288.
K18-hACE2 transgenic mice
SARS-CoV-2 XBB.1 infection model
Oral
25 mg/kg, 50 mg/kg, 100 mg/kg
Treatment started 2 hours post-infection and lasted for 4 days
Evaluate the antiviral efficacy of GZNL-P36 in SARS-CoV-2 infected Mice model, showing significantly improved survival and notable reductions in lung viral loads and lesions.
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