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| Author | Year | Method | Detail of method | Name of compound/drug | Target | Efficacy | Comments |
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| Agarwal et al. [284] | 2020 | In vitro | IL-1β assay using THP-1 cells, in vivo pharmacokinetic investigation in C57BL/6 mice | Alkenyl sulfonylurea derivatives | NLRP3 inhibition and reduction in the release of interleukin-1β | IL-1β inhibition IC50 of 35 nM, good oral absorption showing Cmax of 8.49 μg/mL with an AUC of 48.9 μg·h/mL and terminal half-life of 2.86 h, after oral route of administration at 3 mg/kg dose | Novel thiazolo-alkenyl sulfonylurea derivative 7 as potent, selective, and orally bioavailable NLRP3 inflammasome inhibitor |
| Akaberi et al. [285] | 2020 | In vitro | Vero E6 cells, RT-qPCR for the quantification of viral RNA, expression and purification of SARS-CoV-2 3CL protease in vitro enzymatic assay | NO-Donor S-nitroso-N-acetylpenicillamine (SNAP): nitric oxide | Main protease | Although the viral replication was not completely abolished (at 200 μM and 400 μM), SNAP delayed or completely prevented the development of viral cytopathic effect in treated cells (IC50 = 440.95 μM ± 36.15 SE) | A dose-dependent inhibition of the 3CL protease (400 μM) |
| Bernstein and Zhang [286] | 2020 | In vitro | Vero E6 cells: virus was then added at a multiplicity of infection (MOI) of 0.01, with 1 h allowed for infection. Cytotoxicity: Cell Counting Kit-8 (CCK-8) colorimetric assay | Gallium maltolate (GaM) | Inhibition of viral replication | EC50 (concentration producing 50% inhibition of viral replication) of about 14 μM (CI 95%: 8.9–22.8 μM) | No cytotoxicity was observed at concentrations up to at least 200 μM |
| Bocci et al. [287] | 2020 | In vitro | Ligand-based virtual screening/Vero E6 cells. Uninfected cells and chloroquine as control inhibition of the SARS-CoV-2-mediated CPE following infection in Vero E6 cells tissue culture infectious dose (TCID50) assay | 4000 approved drugs/glafenine, amodiaquine vorinostat, zuclopenthixol, isoxsuprine, nebivolol, ambroxol, panobinostat, and pracinostat | Inhibition of viral replication | In silico: glafenine, amodiaquine (amodiaquine), vorinostat, zuclopenthixol, isoxsuprine, nebivolol, ambroxol, panobinostat, and pracinostat. In vitro: zuclopenthixol and amodiaquine show anti-SARS-CoV-2 activity comparable with chloroquine. EC50 values were estimated as 0.13 μM for AQ, 1.35 μM for ZPX, and 2.72 μM for nebivolol | Zuclopenthixol and nebivolol exhibit in vitro antiviral activities and potencies that are comparable or better than chloroquine and hydroxychloroquine |
| Cao et al. [288] | 2020 | In vitro | African green monkey kidney Vero E6 cells were infected with virus at an MOI of 0.01 for 1 h. Immunofluorescence assay physiologically based pharmacokinetic modeling and simulations | 9 artemisinin-related compounds | Inhibition of viral replication | Arteannuin B showed the highest anti-SARS-CoV-2 potential with an EC (50) of 10.28 ± 1.12 μM. Artesunate and dihydroartemisinin showed similar EC (50) values of 12.98 ± 5.30 μM and 13.31 ± 1.24 μM, respectively | Lumefantrine could inhibit SARS-CoV-2 in vitro with an EC50 of 23.17 ± 3.22 μM. Impair viral infection by modulating host cell metabolic pathways |
| Chen et al. [289] | 2021 | In vitro | Fluorescence-based high-throughput screen, docking/pretreated with various concentrations of test compound for 1 h, followed by infection with SARS-CoV-2 (MOI of 0.0001) in the presence of test compounds, expression and purification of SARS-CoV-2 PLpro, 3CL protease from Escherichia coli enzymatic assay of SARS-CoV-2 PLpro cytotoxicity assay | 1920 natural products | Main protease, papain-like protease | Ginkgolic acid and anacardic acid were also identified as inhibitors of 3CLpro, with IC50 values of 1.79 ± 0.58 and 2.07 ± 0.35 μM, respectively, both ginkgolic acid and anacardic acid dose-dependently inhibited PLpro activity, with IC50 values of 16.30 ± 0.64 and 17.08 ± 1.30 μM, respectively | Ginkgolic acid act as an irreversible inhibitor against both PLpro and 3CLpro, suggesting it is a covalent inhibitor |
| Gendrot et al. [290] | 2020 | In vitro | Vero E6 cells, replication was estimated by RT-PCR/chloroquine as control determination of the inhibition stage | Doxycycline | Inhibition of viral replication | Median effective concentration (EC50) and 90% effective concentration (EC90) for doxycycline were 4.5 ± 2.9 μM and 23.5 ± 16.5 μM, respectively, EC50 = 4.5 μM | Interacted at both entry and postentry stages of the SARS-CoV-2 infection doxycycline has anti-inflammatory effects |
| Gupta et al. [291] | 2021 | In vitro | High-throughput virtual screening, molecular dynamic simulation, prime MM-GBSA/Vero E6 cells infected with the SARS-CoV-2 fluorescence-based biochemical assay for inhibitors of 3CLpro | FDA-approved drugs | Main protease, PLpro, RNA-directed RNA polymerase, helicase, NSP-15, NSP14, and 2′-O-methyltransferase | Troxerutin (main protease and PLpro) and bisindolylmaleimide derivatives (main protease and exon) had possible dual targets. Ivermectin has nonselective toxicity to the ATCC E6 Vero cells at ≤50 μM and 16.67 μM based on the number of nuclei counted | BIM IX specifically blocked 3CLpro in vitro enzymatic assay inhibition was observed with an IC50 value of 113.7 ± 5.2 μM a known inhibitor of protein kinase C isoforms, bisindolylmaleimide IX (BIM IX), was found to be a potent inhibitor of SARS-CoV-2 |
| Hahn et al. [292] | 2020 | In vitro | Vero B4 and 76 cells و Vero E6 cells/remdesivir RT-qPCR for the detection of extracellular SARS-CoV-2 in cell immunostaining for the detection of Intracellular SARS-CoV-2 Western blot analysis viral plaque and yield reduction assay neutral red assay (NRA) | (IMU-838) inhibitor of human dihydroorotate dehydrogenase (DHODH) | Inhibition of viral replication/immunomodulator | A virus-specific EC90 of 6.2 ± 1.9 μM was measured, with no cytotoxicity observed with drug concentrations up to 100 μM at 3 days p.i. | Active moiety of IMU-838, vidofludimus, possesses broad-spectrum antiviral activity. IMU-838 reduces T lymphocyte proliferation, cytokine production, and organ infiltration by leukocytes in various in vivo and in vitro models for autoimmunity |
| Hattori et al. [293] | 2021 | In vitro | VeroE6 cell-based assays with RNA-qPCR, cytopathic assays | Two small-molecule-compounds, named GRL-1720 and 5 h, containing an indoline and indole moiety | Main protease | EC(50) values of 15 ± 4 and 4.2 ± 0.7 μM for GRL-1720 and 5 h | Combination of 5 h and remdesivir exhibits synergism against SARS-CoV-2.5 h might serve as a lead M (pro) inhibitor |
| Jang et al. [294] | 2021 | In vitro | Vero CCL-81 cells/Calu-3 human lung epithelial cells | Gemcitabine and its analog 2′-fluoro-2′-deoxycytidine (2FdC) | Inhibition of viral replication | 50% effective concentration (EC(50)) of 1.2 μM compare to remdesivir 2FdC was marginally active (EC(50) = 175.2 μM) | Gemcitabine has a synergistic effect when combined with remdesivir |
| Jeon et al. [295] | 2020 | In vitro | Drug repositioning/chloroquine, lopinavir, and remdesivir were used as reference drugs/VeroE6 cell/immunofluorescence analysis | 48 FDA-approved drugs | Inhibition of viral replication | Niclosamide (IC50, 0.28 μM), ciclesonide (IC50, 4.33 μM) | 24 potential antiviral drug candidates against SARS-CoV-2 infection |
| Liu et al. [296] | 2020 | In vitro | Virtual screening/spiked pseudotyped lentivirus for imitating SARS-CoV-2 cell entry/ACE2-GFP expressing HEK293T 2 cells. HeLa Cells, 293T (human, kidney) cells, and Vero-E6 (African green monkey, kidney) cells | FDA-approved drugs | Angiotensin I-converting enzyme 2 (ACE2) and a spike protein | Nine potential candidates were selected and submitted to experimental studies. Three (romidepsin, saquinavir, and nelfinavir) of nine drugs possess the ability to suppress SARS-2-S pseudotyped particles to enter the ACE2 expressing cells in a concentration-dependent manner | Five clinical HDAC inhibitors including romidepsin, panobinostat, givinostat hydrochloride monohydrate, CAY10603, and sirtinol could inhibit noticeably the spike/ACE2-mediated cell entry of SARS-CoV-2 |
| Ma et al. [297] | 2020 | In vitro | Vero E6 cells using cytopathic effect and plaque reduction assay. HCoV-229E infection in Huh-7 cells. Western blot assay | Phillyrin (KD-1) ingredient of Forsythia suspensa | Inhibition of viral replication | Inhibit SARS-CoV-2. Markedly reduce the production of pro-inflammatory cytokines (TNF-α, IL-6, IL-1β, MCP-1, and IP-10) at the mRNA levels. p-NF-κB p65, NF-κB p65, and p-IκBα | KD-1 could significantly inhibit virus proliferation in vitro, the upregulated expression of pro-inflammatory cytokines induced by SARS-CoV-2 |
| Masih et al. [298] | 2020 | In vitro | LPS-stimulated RAW267.4 cells by enzyme immunoassay Western blot assay | Pyrazole derivatives | Anti-inflammatory levels of interleukin-1β, tumor necrosis factor-α, and interleukin-6 | Compound 6c as the most potent analog among the tested series. Inhibition of inhibitor kappa B-α and NF-κB | |
| Omotuyi et al. [299] | 2020 | In vitro | Kinetic assay | Aframomum melegueta. 100 secondary metabolites | Main protease, 2′-O-ribose methyltransferase (NSP16), and surface glycoprotein/ACE2 receptor interface | Diarylheptanoid (letestuianin A), phenylpropanoid (4-cinnamoyl-3-hydroxy-spiro[furan-5,2′-(1′H)-indene]-1′,2,3′(2′H,5H)-trione), flavonoids (quercetin, apigenin, and tectochrysin) have been identified as high binding compounds to SARS-CoV-2 targets in a polypharmacology manner | Di-ethyl-ether (IC50 = 0.03 mg/L), acetone (IC50 = 1.564 mg/L), ethyl-acetate (IC50 = 0.382 mg/L), and methanol (IC50 = 0.438 mg/L) fractions demonstrated the best inhibition in kinetic assay |
| Outlaw et al. [300] | 2020 | In vitro | Vero E6 cell β-Gal complementation-based fusion assay. Viral titration and plaque reduction neutralization assay quantitative RT-PCR. | A lipopeptide that is derived from the C-terminal heptad repeat (HRC) domain of SARS-CoV-2 S | Inhibition cell-cell fusion | Inhibits cell-cell fusion mediated by SARS-CoV-2 S and blocks infection by live SARS-CoV-2 in Vero E6 cell monolayers (IC50) of ∼10 nM and 90% inhibitory concentration (IC90) of ∼100 nM | |
| Pitsillou et al. [301] | 2021 | In vitro | Molecular docking, molecular dynamic simulation/papain-like protease (SARS-CoV-2) assay kit GRL-0617 as a control enzymatic inhibition assay | 300 small compounds derived predominantly from our OliveNet™ library (222 phenolics) and supplemented with synthetic and dietary compounds | Papain-like protease | Selection of 30 compounds. Hypericin possessed inhibition activity, and both rutin and cyanidin-3-O-glucoside resulted in a concentration-dependent inhibition of the PLpro | The IC50 values were not clarified in experiment |
| Pizzorno et al. [302] | 2020 | In vitro | Vero E6 cells/remdesivir, lopinavir, chloroquine as comparator | Favipiravir, ribavirin, umifenovir (arbidol), berberine, cyclosporine A, diltiazem | Inhibition of viral replication | Umifenovir, berberine, and cyclosporine A with estimated 50% inhibitory concentrations of 0.99, 5.2, 1.38, 3.5, 10.6, and 3 μM, respectively | A strong antagonism between remdesivir and berberine, in contrast with remdesivir/diltiazem, for which we describe high levels of synergy |
| Plaze et al. [303] | 2021 | In vitro | Monkey VeroE6 cells and human alveolar basal epithelial A549-ACE2 cell RT-qPCR for the presence of SARS-CoV-2 RNA | Chlorpromazine | Inhibition of viral replication | (IC50) of 8.2 μM in monkey VeroE6 cells (CC50) of 13.5 μM, and selectivity index (SI) of 1.65 IC50 of 11.3 μM, CC50 of 23.1 μM, and SI of 2.04 in human A549-ACE2 cells | |
| Reznikov et al. [304] | 2021 | In vitro | Repurposing study molecular docking/human lung A549 cells that were transfected with hACE2 (ACE2-A549) cells, Vero E6 cells | Electronic health records of over 219,000 patients with antihistamines | Inhibition of viral replication/spike protein | IC50 15.3 μg/ml for hydroxyzine, 17.4 μg/ml for diphenhydramine, and 2.24 μg/ml for azelastine | Hydroxyzine and possibly azelastine bind angiotensin-converting enzyme-2 (ACE2) and the sigma-1 receptor as off-target disease prevention |
| Sakurai et al. [305] | 2021 | In vitro | VeroE6 cells/human colon-derived Caco-2 cells/infection assay with immunofluorescence | 5-Amino levulinic acid (5-ALA) | Inhibition of viral replication | IC50 of 570 μM/IC50 of 63 μM in human Caco-2 cells. No significant cytotoxicity | 5-ALA is synthesized in most animals and plants and we are continuously consuming it in our food. Show anti-inflammation effects in humans |
| Stone et al. [306] | 2021 | In vitro | Vero E6 monkey kidney and Calu-3 human lung adenocarcinoma cells/RT-qPCR of RNA | Stenoparib poly(ADP ribose) polymerase (PARP) inhibitor | Inhibition of viral replication | Dose-dependent activity against SARS-CoV-2 at concentrations up to 30 μM with negligible cytotoxicity. Synergistic effect with remdesivir | Stenoparib impedes entry and postentry processes |
| Su et al. [307] | 2020 | In vitro | Vero E6 cells, quantitative real-time RT-PCR | Shuanghuanglian preparation, a Chinese traditional patent medicine (chlorogenic acid, phillyrin, baicalin, and baicalein) | Main protease | Dose-dependent inhibition of SARS-CoV-2 3CLpro, and the resulting IC50 values were 0.090, 0.064, and 0.076 μL/mL for three Shuanghuanglian oral liquids produced by three different pharmaceutical companies | Baicalin and baicalein, which have good drug-like properties |
| Touret et al. [308] | 2020 | In vitro | VeroE6 cells/human colon-derived Caco-2 cell real-time RT-PCR/remdesivir as control | FDA-approved chemical library | Inhibition of viral replication | 23 drugs were selected to cover the 12 different groups. 11 compounds such as macrolide antibiotics, proton pump inhibitors, antiarrhythmic agents, or CNS drugs emerged showing antiviral potency with 2 < EC50 ≤ 20 μM | Two of the highest antiviral activity were obtained for azithromycin (EC50 = 2.12 μM) and hydroxychloroquine (EC50 = 4.17 μM). Vonoprazan and sulfadoxine show a moderate activity with EC50 above 30 μM |
| Tree et al. [309] | 2021 | In vitro | Plaque inhibition assay with Vero E6 cells/differential scanning fluorimetry/ELISA assays | Unfractionated heparin (UFH), low MW heparins | Inhibition of viral replication/spike protein RBD | All the UFH preparations had potent antiviral effects, with IC50 values ranging between 25 and 41 μg·ml−1, whereas LMWHs were less inhibitory by ∼150-fold (IC50 range 3.4–7.8 mg·ml−1) | Heparin directly inhibits the binding of RBD to the human ACE2 protein receptor |
| Vatansever et al. [310] | 2021 | In vitro | Docking, screening assay/Vero E6, and A549 cell culture | 30 FDA/EMA-approved drugs | Main protease | Six FDA/EMA-approved drugs can potently inhibit Mpro with an IC50 value lower than 100 μM. Bepridil exhibited strong inhibition of SARS-CoV-2 from entry and replication inside Vero E6 and A549 cells (IC50 = 72 μM) | Bepridil indicated that it had low micromolar EC50 values |
| Wang et al. [311] | 2020 | In vitro | FRET-based enzyme activity assay of Mpro measurement of human TMPRSS2 activity by a FRET-based enzymatic assay surface plasmon resonance (SPR) analysis molecular docking African green monkey kidney cell Vero E6 | Catechin, kaempferol, quercetin, proanthocyanidins, resveratrol, and tannic acid | TMPRSS2 (transmembrane protease serine 2) main protease | Tannic acid inhibited the activities of the two proteases with an IC50 of 13.4 mM for Mpro and 2.31 mM for TMPRSS2 | A potent dual inhibitor |
| Watashi [312] | 2021 | In vitro | Virtual drug screening, docking/VeroE6 cells detecting the viral RNA by real-time RT-PCR or viral proteins by immunofluorescence | FDA-approved drugs | Main protease, PLpro, RdRp, helicase, spike, ACE2, and TMPRSS2 | Abiraterone, amodiaquine, anidulafungin, arbidol, astemizole, atazanavir, auranofin, azithromycin bazedoxifene, bexarotene, camostat IC50 values <5 μM | Cepharanthine, cetilistat, chloroquine, ciclesonide, cyclosporine A, digoxin, ivermectin, lopinavir, mefloquine, nelfinavir, niclosamide |
| Yu et al. [313] | 2021 | In vitro | Computer-aided drug design and biological verification surface plasmon resonance (SPR) assays and NanoBiT assays | Traditional Chinese medicine | Spike proteins | Glycyrrhizic acid, the most efficient and nontoxic broad-spectrum anticoronavirus molecule IC50, was 22 μM | |
| Zandi et al. [314] | 2021 | In vitro | African green monkey kidney cells (Vero CCL-81 cells) and reverse transcription-quantitative PCR (qRT-PCR). Remdesivir and β-D-N4-hydroxycytidine as positive drug controls | Nucleoside analogs/anti-HCV agents | Inhibition of viral replication | Sofosbuvir and favipiravir demonstrated no antiviral effect against COVID-2 | |
| Zhang et al. [315] | 2020 | In vitro | Hybrid virtual screening, docking, force field-based simulation/qRT-PCR assay, indirect immunofluorescence assay (IFA) and CCK-8 assay, surface plasmon resonance (SPR) assay/Vero cells | 1906 approved drugs from TargetMol-approved drug library | RNA-dependent RNA polymerase (RdRp) | Pralatrexate and azithromycin (EC50) values of 0.008 and 9.453 μM | A new therapeutic agent pralatrexate against COVID-19 by targeting RdRp |
| Zhang et al. [316] | 2020 | In vitro | African green monkey kidney Vero E6 cell line, quantitative real-time PCR (RT-PCR) analysis, immunofluorescence microscopy, Western blot analysis | Lopinavir/ritonavir, rupintrivir, and AG7404 | Inhibition of viral replication | (IC50) and half cytotoxic concentration (CC50) values of lopinavir were ∼12.01 μmol/L and 80.82 μmol/L; for ritonavir, the IC50 and CC50 values were 19.88 μmol/L and 94.71 μmol/L (SI = 4.76) | Rupintrivir inhibited SARS-CoV-2 infection only at high drug concentrations (Vero E6: IC50 = 34.08 μmol/L, AG7404: Vero E6: IC50∼195.8 μmol/L). Lopinavir/ritonavir should be stopped for clinical use due to the huge gap between in vitro IC50 and free plasma concentration |
| Zhu et al. [317] | 2021 | In vitro | Human fibroblast lung cells (MRC-5), cytopathic effect (CPE) assay, primary human airway air-liquid interface (ALI) cultures, TEER assay, CCK-8 assays | Stimulator of interferon genes (STING): dimeric amidobenzimidazole (diABZI) | Inhibition of viral replication | Potent anticoronavirus activity against both the common cold human coronavirus 229E (HCoV-229E) and SARS-CoV-2 in cell culture systems. EC50 of 120 μM | |
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