Journal: Cell Reports

Article Title: SARS-CoV-2 variant B.1.617 is resistant to bamlanivimab and evades antibodies induced by infection and vaccination

doi: 10.1016/j.celrep.2021.109415

Figure Lengend Snippet: Entry driven by the S protein of SARS-CoV-2 variant B.1.617 can be blocked with soluble ACE2 and camostat mesylate (A) S protein-bearing particles were incubated with different concentrations of soluble ACE2 (sol-ACE2) for 30 min at 37°C before the mixtures were inoculated onto Caco-2 cells. (B) Caco-2 target cells were preincubated with different concentrations of the serine protease inhibitor camostat mesylate for 1 h at 37°C before S protein-bearing particles were added. Transduction efficiency was quantified by measuring virus-encoded luciferase activity in cell lysates 16–18 h after transduction. For normalization, SARS-CoV-2 S protein-driven entry in the absence of sol-ACE2 or camostat was set as 0% inhibition. Presented are the average (mean) data from three biological replicates (each performed with technical quadruplicates. Error bars indicate the SEM. Statistical significance of differences between the WT and the variant S proteins or VSV-G was analyzed by two-way ANOVA with Dunnett’s posttest (p > 0.05, ns [not significant, not indicated in the graphs]; ∗∗∗ p ≤ 0.001).

Article Snippet: In order to study the antiviral activity of Camostat mesylate, Caco-2 target cells, which express endogenous TMPRSS2, were pre-incubated for 1 h with medium containing different concentrations (100, 10, 1, 0.1 or 0.01 μM) of Camostat mesylate (prepared from a 100 mM stock; Tocris) or the solvent dimethyl sulfoxide (DMSO, 1:1,000; Sigma-Aldrich) as control before being inoculated with VSV bearing S protein, VSV-G or no glycoprotein.

Techniques: Variant Assay, Incubation, Protease Inhibitor, Transduction, Virus, Luciferase, Activity Assay, Inhibition

Journal: Journal of Clinical Investigation

Article Title: Trypsin activity governs increased susceptibility to pancreatitis in mice expressing human PRSS1R122H

doi: 10.1172/jci130172

Figure Lengend Snippet: Figure 7. Effective experimental therapeutics with an FDA- approved anticoagulation agent in PRSS1R122H mice. (A) Schema of pancreatitis induction and treatment in PRSS1R122H mice. Pancreatitis was induced by cerulein, and therapeutic drugs were administered 5 hours after the first cerulein injection. (B) After 7 days of treatment, pancreata in the untreated control group became smaller. In con- trast, pancreata in dabigatran-treated groups were mostly normal. Camostat only exhibited intermediate effects at higher does (300 mg/kg). Both drugs were given twice daily for 7 days. Mean ± SEM (n = 10 per group). ***P < 0.001; 1-way ANOVA with Tukey’s multiple comparisons test. (C) Histology score evaluation showed that dabig- atran significantly improved CP. Mean ± SEM (n = 10 per group). ***P < 0.001; 1-way ANOVA with Tukey’s multiple comparisons test. (D) Representative images of H&E staining after drug treatments over 7 days (n = 10 per group). Scale bars: 200 μm.

Article Snippet: For therapeutic studies, mice were randomly assigned into groups and treated by oral administration with vehicle (25% Captisol in normal saline; catalog RC-0C7-K01; Ligand Pharmaceuticals Inc.) or camostat mesylate (catalog sc-203867; Santa Cruz), dabigatran etexilate mesylate (catalog D100150; Toronto Research Chemicals), apixaban (catalog QC-7598-001; Combi Blocks), Rivaroxaban (catalog QA-2934-001; Combi Blocks), or by i.p. injection with Argatroban (catalog A769000; Toronto Research Chemicals).

Techniques: Injection, Control, Staining

Journal: Journal of Clinical Investigation

Article Title: Trypsin activity governs increased susceptibility to pancreatitis in mice expressing human PRSS1R122H

doi: 10.1172/jci130172

Figure Lengend Snippet: Figure 8. Anticoagulation and trypsin inhibition synergistically improved pancreatitis in PRSS1R122H mice. (A) Immunohistochemical analysis showed intrapancreatic fibrin deposition, an indicator of increased coagulation, in the pancreata of PRSS1R122H mice 24 hours after cerulein induction (n = 5). Scale bars: 200 μm. (B) Western blot showed that active thrombin significantly increased in the pancreata of PRSS1R122H mice compared with those from C57BL/6J mice. Representative blots from 3 independent experiments are shown. (C) Schema of pancreatitis induction and drug treatment with the anticoagulation specific agent (apixaban), trypsin specific inhibitor (camostat), or in combination. Starting 5 hours after pancreatitis induction, drugs were given twice daily by oral gavage over 7 days. (D) Combination of anticoagulation therapy with factor Xa inhibitor apixaban (100 mg/kg) and trypsin inhibitor camostat (200 mg/kg) greatly protected the pancreas, as manifested by preservation of pancreas mass. Mean ± SEM (n = 15 per group). ***P < 0.001; 1-way ANOVA with Tukey’s multiple comparisons test. (E) Representative histologic images of H&E staining from untreated, apixaban alone, camostat alone, and combination therapy–treated mice (n = 15 per group). Scale bars: 200 μm. (F) Overall histology score evaluation of the mice. Mean ± SEM. Representative results from 6 mice per group are shown. ***P < 0.001; 1-way ANOVA with Tukey’s multiple comparisons test.

Article Snippet: For therapeutic studies, mice were randomly assigned into groups and treated by oral administration with vehicle (25% Captisol in normal saline; catalog RC-0C7-K01; Ligand Pharmaceuticals Inc.) or camostat mesylate (catalog sc-203867; Santa Cruz), dabigatran etexilate mesylate (catalog D100150; Toronto Research Chemicals), apixaban (catalog QC-7598-001; Combi Blocks), Rivaroxaban (catalog QA-2934-001; Combi Blocks), or by i.p. injection with Argatroban (catalog A769000; Toronto Research Chemicals).

Techniques: Inhibition, Immunohistochemical staining, Coagulation, Western Blot, Preserving, Staining

Journal: AAPS PharmSciTech

Article Title: Formulation and In-Vitro Testing of Nebulized Camostat Mesylate Loaded Nanoliposomes for the Treatment of SARS-CoV- 2 Infection.

doi: 10.1208/s12249-025-03099-3

Figure Lengend Snippet: Fig. 6 A plot of calculated concentrations as a function of known concentrations of Camostat Mesylate using the HPLC method

Article Snippet: Camostat Mesylate (Lot# B20BS04231) was obtained from BOC Sciences in Shirley, New York, USA.

Techniques:

Journal: bioRxiv

Article Title: SARS-CoV-2 variants with mutations at the S1/S2 cleavage site are generated in vitro during propagation in TMPRSS2-deficient cells

doi: 10.1101/2020.08.28.271163

Figure Lengend Snippet: (a) Vero-TMPRSS2 cells were infected with SARS-CoV-2 WT or del2 mutant in the presence of varying concentrations of the TMPRSS2 inhibitor, camostat, or the cathepsin B/L inhibitor, E-64d, for 1h. At 6 h post-inoculation, the relative levels of viral N protein RNA were evaluated quantitatively by qRT-PCR. (b) Vero-TMPRSS2 and Vero cells were infected with SARS-CoV-2 WT or S gene mutants in the presence of 50 μM camostat and/or 25 μM E-64d for 1 h. At 6 h post-inoculation, the relative levels of viral N protein RNA were quantified by qRT-PCR. Cellular β-actin mRNA levels were used as reference controls. The values shown are mean ± SD of triplicate samples. One-way analysis of variance with Dunnett’s test was used to determine the statistical significance between the responses to treatment with inhibitors and the no-treatment controls; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Cells were infected with either WT or S mutants of SARS-CoV-2 at an MOI of 0.1 in the presence of 50 μM camostat mesylate (FUJIFILM Wako Pure chemical) and/or 25 μM E-64d (Abcam) for 1 h. Cells were then washed twice with PBS and cultured in fresh culture medium with 2% FBS.

Techniques: Infection, Mutagenesis, Quantitative RT-PCR

Journal: bioRxiv

Article Title: SARS-CoV-2 Spike evolution influences GBP and IFITM sensitivity

doi: 10.1101/2022.03.07.481785

Figure Lengend Snippet: Inhibition of spike-mediated PV infection of Caco2 cells by A) Camostat and B) E64d inhibitors. Shown is a representative titration. C) Inhibition of spike PV infection Caco2 cells in the presence of 25 μM Camostat or 25 μM E64d normalised to DMSO control. Infection was measured by luciferase assay. D) Inhibition of SARS-CoV-2 live virus infection of Caco2 cells in presence of 25 μM Camostat or 25 μM E64d. Infection was measured by flow cytometry staining for nucleoprotein and shown as the percentage N-positive cells (% N+). Left panel shows percentage N+ cells at 24 hpi, and the right panel shows the percentage infection for each virus normalised to the corresponding DMSO control. E) Immunoblot analysis of spike cleavage in SARS-CoV-2 virions. Indicated virus isolates were purified from culture supernatants and immunoblotted for spike and nucleoprotein. Band quantification shows relative proportion of cleaved spike (S2/S) and cleaved spike incorporation into virions (S2/N). F-H) Effect of GBPs on SARS-CoV-2 spike-mediated entry pathways. Wuhan-Hu-1 spike PV made in presence of 80 ng of indicated GBP plasmid or EV control was used to infect Caco2 cells in the presence of increasing concentrations of F) Camostat or G) E64d. Data show percentage infection compared to DMSO EV control. H) Inhibition of Wuhan-Hu-1 PV +/-GBPs and in the presence of 25 μM Camostat or 25 μM E64d. Data are normalised to DMSO control separately for each GBP treatment and grouped by inhibitor treatment. I-J) Omicron spike PV was made in presence of GBP5 or EV control and infection of Caco2 cells in the presence of increasing doses of (I) Camostat or (J) E64d. Data show the mean ±SEM from three independent experiments. Groups were compared using a two-way ANOVA with Dunnett’s multiple comparisons test. Ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Article Snippet: For inhibitor studies cells were pre-treated before infections for 2 h with Camostat mesylate (Apexbio, 0.2-100 μM) or E64d (Focus Biomolecules, 0.2-25 μM).

Techniques: Inhibition, Infection, Titration, Luciferase, Flow Cytometry, Staining, Western Blot, Purification, Plasmid Preparation

Journal: bioRxiv

Article Title: SARS-CoV-2 Spike evolution influences GBP and IFITM sensitivity

doi: 10.1101/2022.03.07.481785

Figure Lengend Snippet: Inhibition of spike-mediated PV infection of HeLa-ACE2 cells by A) Camostat and B) E64d inhibitors. Shown is a representative titration. C) Inhibition of spike PV infection of HeLa-ACE2 cells in the presence of 25 μM Camostat or 25 μM E64d normalised to DMSO control. Infection was measured by luciferase assay. D) Inhibition of SARS-CoV-2 live virus infection of HeLa-ACE2 cells in presence of 25 μM Camostat or 25 μM E64d. Infection was measured by flow cytometry staining for nucleoprotein and shown as the percentage N-positive cells (% N+). Left panel shows percentage N+ cells at 24 hpi, and the right panel shows the percentage infection for each virus normalised to the corresponding DMSO control. Shown is mean ±SEM from three independent experiments. Groups were compared using a two-way ANOVA with Dunnett’s multiple comparisons test. Ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Article Snippet: For inhibitor studies cells were pre-treated before infections for 2 h with Camostat mesylate (Apexbio, 0.2-100 μM) or E64d (Focus Biomolecules, 0.2-25 μM).

Techniques: Inhibition, Infection, Titration, Luciferase, Flow Cytometry, Staining

Journal: bioRxiv

Article Title: SARS-CoV-2 Spike evolution influences GBP and IFITM sensitivity

doi: 10.1101/2022.03.07.481785

Figure Lengend Snippet: A) Expression of HA-tagged IFITMs in Caco2 cells stably expressing IFITM1-3 as measured by flow cytometry. Left panel shows a representative histogram. B) Immunofluorescence images of stable Caco2 cells expressing IFITM1-3. HA-tag IFITM shown in green, CD63 shown in red, and nuclei shown in blue. Scale bar is 50 μm. C-G) Caco2 cells expressing IFITMs were infected with live SARS-CoV-2 virus. Shown is percent N+ cells after 24 hpi of C) VIC, D) IC19, E) Alpha, F) Delta, and G) Omicron isolates. H-L) PV infection of IFITM transduced Caco2 cells with H) Wuhan-Hu-1, I) Wuhan-Hu-1 D614G, J) Alpha, K) Delta, and L) Omicron spike PV. Data are shown as percent infectivity normalised to WT Caco2 cells (no IFITM over-expression). M) Caco2 cells expressing IFITMs were incubated with Camostat inhibitor (5 μM) or DMSO control and infected with Wuhan-Hu-1 PV. Percent infectivity was determined by normalising to WT Caco2 cells DMSO control. N-R) IFITM expressing Caco2 cells were infected with spike PVs made in presence of GBP5 or EV control. Data show the infectivity normalised to EV control on WT cells for PV made using N) Wuhan-Hu-1, O) Wuhan-Hu-1 D614G, P) Alpha, Q) Delta, and R) Omicron spikes. Data show the mean ±SEM from three independent experiments. Groups were compared A-L) using a one-way ANOVA or M-R) using a two-way ANOVA with Dunnett’s multiple comparisons test. Ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Article Snippet: For inhibitor studies cells were pre-treated before infections for 2 h with Camostat mesylate (Apexbio, 0.2-100 μM) or E64d (Focus Biomolecules, 0.2-25 μM).

Techniques: Expressing, Stable Transfection, Flow Cytometry, Immunofluorescence, Infection, Over Expression, Incubation

Journal: mBio

Article Title: Drug Combinations as a First Line of Defense against Coronaviruses and Other Emerging Viruses

doi: 10.1128/mbio.03347-21

Figure Lengend Snippet: Drugs reported to synergize with remdesivir or molnupiravir to inhibit SARS-CoV-2 infection of Calu3 lung cells ^a

Article Snippet: , Camostat , Preclin. , Ph3 , Yes , TMPRSS2 , Fusion , .

Techniques: Infection

Journal: NPJ Digital Medicine

Article Title: Executable network of SARS-CoV-2-host interaction predicts drug combination treatments

doi: 10.1038/s41746-022-00561-5

Figure Lengend Snippet: List of approved drugs used for in silico screening and their targets in the network model.

Article Snippet: Caco-2 cells were pre-treated with Camostat Mesilate (ApexBio, B2082), Apilimod (Selleckchem, S6414), Miltefosine (Cayman Chemicals, 63280), Dexamethasone (EMD Millipore, 265005) or DMSO (Sigma) vehicle control at 2x the final concentration for 2 h prior to infection in 50 μl culture medium.

Techniques: In Silico, Glycoproteomics, Recombinant

Journal: NPJ Digital Medicine

Article Title: Executable network of SARS-CoV-2-host interaction predicts drug combination treatments

doi: 10.1038/s41746-022-00561-5

Figure Lengend Snippet: a Percentage cells expressing nucleocapsid protein relative to control for Apilimod and Camostat combinations. b Percentage cells expressing nucleocapsid protein for control, Apilimod (10 µM), Camostat (100 µM) or a combination of Apilimod (10 µM) and Camostat (100 µM). Data show the mean and standard error, from three independent experiments. *** indicates p < 0.001 by two-way ANOVA.

Article Snippet: Caco-2 cells were pre-treated with Camostat Mesilate (ApexBio, B2082), Apilimod (Selleckchem, S6414), Miltefosine (Cayman Chemicals, 63280), Dexamethasone (EMD Millipore, 265005) or DMSO (Sigma) vehicle control at 2x the final concentration for 2 h prior to infection in 50 μl culture medium.

Techniques: Expressing, Control

Journal: NPJ Digital Medicine

Article Title: Executable network of SARS-CoV-2-host interaction predicts drug combination treatments

doi: 10.1038/s41746-022-00561-5

Figure Lengend Snippet: Summary of predicted effective treatments for early and late severe COVID-19.

Article Snippet: Caco-2 cells were pre-treated with Camostat Mesilate (ApexBio, B2082), Apilimod (Selleckchem, S6414), Miltefosine (Cayman Chemicals, 63280), Dexamethasone (EMD Millipore, 265005) or DMSO (Sigma) vehicle control at 2x the final concentration for 2 h prior to infection in 50 μl culture medium.

Techniques: Recombinant