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20 pages, 4529 KiB

Open AccessArticle

Plant Metabolites as SARS-CoV-2 Inhibitors Candidates: In Silico and In Vitro Studies

by Alberto Jorge Oliveira Lopes, Gustavo Pereira Calado, Yuri Nascimento Fróes, Sandra Alves de Araújo, Lucas Martins França, Antonio Marcus de Andrade Paes, Sebastião Vieira de Morais, Cláudia Quintino da Rocha and Cleydlenne Costa Vasconcelos

Pharmaceuticals 2022, 15(9), 1045; https://doi.org/10.3390/ph15091045 - 24 Aug 2022

Cited by 15 | Viewed by 2804

Abstract

Since it acquired pandemic status, SARS-CoV-2 has been causing all kinds of damage all over the world. More than 6.3 million people have died, and many cases of sequelae are in survivors. Currently, the only products available to most of the world’s population [...] Read more.

Since it acquired pandemic status, SARS-CoV-2 has been causing all kinds of damage all over the world. More than 6.3 million people have died, and many cases of sequelae are in survivors. Currently, the only products available to most of the world’s population to fight the pandemic are vaccines, which still need improvement since the number of new cases, admissions into intensive care units, and deaths are again reaching worrying rates, which makes it essential to compounds that can be used during infection, reducing the impacts of the disease. Plant metabolites are recognized sources of diverse biological activities and are the safest way to research anti-SARS-CoV-2 compounds. The present study computationally evaluated 55 plant compounds in five SARS-CoV-2 targets such Main Protease (Mpro or 3CL or MainPro), RNA-dependent RNA polymerase (RdRp), Papain-Like Protease (PLpro), NSP15 Endoribonuclease, Spike Protein (Protein S or Spro) and human Angiotensin-converting enzyme 2 (ACE-2) followed by in vitro evaluation of their potential for the inhibition of the interaction of the SARS-CoV-2 Spro with human ACE-2. The in silico results indicated that, in general, amentoflavone, 7-O-galloylquercetin, kaempferitrin, and gallagic acid were the compounds with the strongest electronic interaction parameters with the selected targets. Through the data obtained, we can demonstrate that although the indication of individual interaction of plant metabolites with both Spro and ACE-2, the metabolites evaluated were not able to inhibit the interaction between these two structures in the in vitro test. Despite this, these molecules still must be considered in the research of therapeutic agents for treatment of patients affected by COVID-19 since the activity on other targets and influence on the dynamics of viral infection during the interaction Spro x ACE-2 should be investigated. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Figure 1
Schematic representation of the four plant metabolites that presented the most promising interaction parameters with SARS-CoV-2 drug targets, through molecular docking. Full article ">Figure 2
The surface representation of plant metabolites docking positions on the SARS-CoV-2 Mpro active site is shown, with amentoflavone (green), kaempferitrin (blue), 7-O-galloylquercetin (yellow), and gallagic acid (magenta) (A). Contacts of SARS-CoV-2 Mpro active site residues with amentoflavone (B), 7-O-galloylquercetin (C), kaempferitrin (D), and gallagic acid (E) are depicted in a two-dimensional diagram. Dashed black lines indicate hydrogen bonds; full green lines indicate van der Waals interactions. Full article ">Figure 3
Surface representation of plant metabolites docking positions on the SARS-CoV-2 RdRp active site, with amentoflavone (green), kaempferitrin (blue), 7-O-galloylquercetin (yellow), and gallagic acid (magenta) (A). The two-dimensional diagram from contacts of SARS-CoV-2 RDRP active site residues with amentoflavone (B), 7-O-galloylquercetin (C), kaempferitrin (D) and gallagic acid (E) Dashed black lines indicate hydrogen bonds; full green lines indicate van der Waals interactions. Full article ">Figure 4
Amentoflavone (green), kaempferitrin (blue), and 7-O-galloylquercetin (yellow) docking positions on the SARS-CoV-2 Papain-like protease (PLpro) active site are shown on the surface (A). Contacts of SARS-CoV-2 PLpro active site residues with amentoflavone (B), 7-O-galloylquercetin (C), and kaempferitrin (D) are depicted in a two-dimensional diagram. Dashed black lines represent hydrogen bonds; full green lines represent van der Waals interactions; and dashed green lines represents π-π stacking. Full article ">Figure 5
Surface representation of plant metabolites docking positions on the SARS-CoV-2 NSP15 endoribonuclease active site, with amentoflavone (green), 7-O-galloylquercetin (yellow), and gallagic acid (magenta) (A). Contacts of SARS-CoV-2 NSP15 endoribonuclease active site residues with gallagic acid (B), amentoflavone (C), and 7-O-galloylquercetin (D) are depicted in two dimensions. Dashed black lines represent hydrogen bonds, while full green lines represent van der Waals interactions. dashed green line represent π-π stacking. Full article ">Figure 6
Surface representation of plant metabolites docking positions on the SARS-CoV-2 Spike protein (Spro) receptor-binding domain (RBD) active site, with amentoflavone (green), kaempferitrin (blue), and gallagic acid (magenta) (A). Contacts of SARS-CoV-2 Spike protein active site residues with amentoflavone (B), gallagic acid (C), and kaempferitrin (D) are depicted in a two-dimensional diagram. Dashed black lines represent hydrogen bonds, while full green lines represent van der Waals interactions, dashed green line represent π-π stacking. Full article ">Figure 7
Amentoflavone (green), 7-O-galloylquercetin (yellow), and gallagic acid (magenta) docking positions on human ACE-2 (SARS-CoV-2 RBD spike protein binding site), in surface view (A). Contacts of SARS-CoV-2 Spike protein active site residues with amentoflavone (B), gallagic acid (C), and kaempferitrin (D) are depicted in a two-dimensional diagram. Dashed black lines indicate hydrogen bonds; full green lines indicate van der Waals interactions. Full article ">

21 pages, 4022 KiB

Open AccessArticle

Efficacy of Kan Jang^® in Patients with Mild COVID-19: Interim Analysis of a Randomized, Quadruple-Blind, Placebo-Controlled Trial

by Levan Ratiani, Elene Pachkoria, Nato Mamageishvili, Ramaz Shengelia, Areg Hovhannisyan and Alexander Panossian

Pharmaceuticals 2022, 15(8), 1013; https://doi.org/10.3390/ph15081013 - 17 Aug 2022

Cited by 7 | Viewed by 3726

Abstract

Kan Jang^®, the fixed combination of Andrographis paniculata (Burm. F.) Wall. ex. Nees and Eleutherococcus senticosus (Rupr. & Maxim.) Maxim extracts, is a herbal medicinal product for relieving symptoms of upper respiratory tract infections. This study aimed to assess the efficacy [...] Read more.

Kan Jang^®, the fixed combination of Andrographis paniculata (Burm. F.) Wall. ex. Nees and Eleutherococcus senticosus (Rupr. & Maxim.) Maxim extracts, is a herbal medicinal product for relieving symptoms of upper respiratory tract infections. This study aimed to assess the efficacy of Kan Jang^®/Nergecov® on duration and the relief of inflammatory symptoms in adults with mild COVID-19. 86 patients with laboratory-confirmed COVID-19 and mild symptoms for one to three days received supportive treatment (paracetamol) and six Kan Jang^® (daily dose of andrographolides—90 mg) or placebo capsules a day for 14 consecutive days in this randomized, quadruple-blinded, placebo-controlled, two-parallel-group study. The primary efficacy outcomes were the decrease in the acute-phase duration and the severity of symptoms score (sore throat, runny nose, cough, headache, fatigue, loss of smell, taste, pain in muscles), an increase in cognitive functions, physical performance, quality of life, and decrease in IL-6, c-reactive protein, and D-dimer in blood. Kan Jang^®/Nergecov^® was effective in reducing the risk of progression to severe COVID-19, decreasing the disease progression rate by almost 2.5-fold compared to placebo. Absolute risk reduction by Kan Jang treatment is 14%, the relative risk reduction is 243.9%, and the number Needed to Treat is 7.14. Kan Jang^®/Nergecov^® reduces the duration of disease, virus clearance, and days of hospitalization and accelerates recovery of patients, relief of sore throat, muscle pain, runny nose, and normalization of body temperature. Kan Jang^®/Nergecov^® significantly relieves the severity of inflammatory symptoms such as sore throat, runny nose, and muscle pain, decreases pro-inflammatory cytokine IL-6 level in the blood, and increases patients’ physical performance (workout) compared to placebo. In this study, for the first time we demonstrate that Kan Jang^®/Nergecov^® is effective in treating mild COVID-19. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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16 pages, 6112 KiB

Open AccessArticle

Search for Novel Potent Inhibitors of the SARS-CoV-2 Papain-like Enzyme: A Computational Biochemistry Approach

by Manuel I. Osorio, Osvaldo Yáñez, Mauricio Gallardo, Matías Zuñiga-Bustos, Jorge Mulia-Rodríguez, Roberto López-Rendón, Olimpo García-Beltrán, Fernando González-Nilo and José M. Pérez-Donoso

Pharmaceuticals 2022, 15(8), 986; https://doi.org/10.3390/ph15080986 - 11 Aug 2022

Cited by 4 | Viewed by 2334

Abstract

The rapid emergence and spread of new variants of coronavirus type 2, as well as the emergence of zoonotic viruses, highlights the need for methodologies that contribute to the search for new pharmacological treatments. In the present work, we searched for new SARS-CoV-2 [...] Read more.

The rapid emergence and spread of new variants of coronavirus type 2, as well as the emergence of zoonotic viruses, highlights the need for methodologies that contribute to the search for new pharmacological treatments. In the present work, we searched for new SARS-CoV-2 papain-like protease inhibitors in the PubChem database, which has more than 100 million compounds. Based on the ligand efficacy index obtained by molecular docking, 500 compounds with higher affinity than another experimentally tested inhibitor were selected. Finally, the seven compounds with ADME parameters within the acceptable range for such a drug were selected. Next, molecular dynamics simulation studies at 200 ns, ΔG calculations using molecular mechanics with generalized Born and surface solvation, and quantum mechanical calculations were performed with the selected compounds. Using this in silico protocol, seven papain-like protease inhibitors are proposed: three compounds with similar free energy (D28, D04, and D59) and three compounds with higher binding free energy (D60, D99, and D06) than the experimentally tested inhibitor, plus one compound (D24) that could bind to the ubiquitin-binding region and reduce the effect on the host immune system. The proposed compounds could be used in in vitro assays, and the described protocol could be used for smart drug design. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Full article ">Figure 1
Diagram of the selection and evaluation of PLpro inhibitor compounds. Compound selection and evaluation process (green background). * PLpro inhibitor, in vitro tested, designed by chemical modification of GRL0617 [<a href="#B17-pharmaceuticals-15-00986" class="html-bibr">17</a>]. Full article ">Figure 2
(A–I) Bioavailability radar of the selected compounds. The red area represents the optimal range for each property (lipophilicity: XLOGP3 between −0.7 and +5.0, size: MW between 150 and 500 g/mol, polarity: TPSA between 20 and 130 Å2, solubility: log S not greater than 6, saturation: fraction of carbons in sp3 hybridization, not less than 0.25, and flexibility: no more than 9 rotatable bonds). Under each bioavailability radar, the 2D chemical structure of each compound is observed. Full article ">Figure 3
Conformations of the PLpro/ligand complex obtained by MD simulation. Crystallographic models of PLpro inhibitors GRL0617 and 12C (A and B, respectively) and seven new inhibitor compounds (D24, D28, D04, D59, D60, D99, and D06) obtained by in the in silico search and molecular docking (C–I) were used to perform 200 ns MD simulations (A,B,D–I). For compounds C12 (B), D24 (C), D28 (D), D04 (E), D59 (F), D60 (G), D99 (H), and D06 (I), an increase in the binding region of each ligand is observed. The red sphere corresponds to the Zn2+ ion. The position of the compounds is observed every ns (red 0 ns and blue 200 ns) of the simulation after superimposing the polypeptide chains. For compound D24 (C), 400 ns of simulation are observed (red 0 ns and blue 400 ns). The structural formula of each compound is shown in the <a href="#app1-pharmaceuticals-15-00986" class="html-app">Supplementary Materials (Figure S3)</a>. Full article ">Figure 4
Residues of less than 3 Å (A–I) of the ligands and the perturbation they generate in the protein structure (J) during MD simulation. Simulations of 200 ns of GRL0617, 12C, D28, D04, D59, D60, D99, and D06 (A, B, D, E, F, G, and I, respectively) and 400 ns for D24 (C) were analyzed, showing the percentage of the simulation in which each ligand (blue 20% to red 100%) was found within 3 Å of the ligand. The perturbation of the protein structure (RMSF) was analyzed during the last 150 ns of each simulation and compared to the protein without ligand or Apo (black dots). In the figure, the region of greatest perturbation is marked (rectangle with a dotted line), and the region affected by the perturbation (red) and the ligand binding site (green circle) are highlighted on the protein. Full article ">Figure 5
Fraction of intermolecular hydrogen bonds for SARS-CoV-2 PLpro interacting with controls (A,B) and selected ligands (C–I). The bar graph shows the most common hydrogen bonds formed between the pocket residues and the studied molecules. Values obtained from the CPPTRAJ script in AMBER. Full article ">Figure 6
Non-covalent interactions in the representative conformation of the PLpro/ligand complex obtained by cluster analysis. The amino acids surrounding with controls GRL0617 and 12C (A and B, respectively) and selected ligands D24, D28, D04, D59, D60, D99, and D06 (C, D, E, F, G, H, and I, respectively) in the PLpro binding pocket are highlighted (upper figures), and in the two-dimensional PLpro/ligand interaction map (lower figures) the NCIPLOT isosurface gradient (0.5 au) is highlighted. Dashed lines indicate possible interactions between amino acids and adjacent ligands. Full article ">Figure 7
Free energy of binding (ΔGbind) of controls (GRL0617 and 12C) and selected compounds (D24, D28, D04, D59, D60, D99, and D06) to SARS-CoV-2 PLpro. ΔGbind was calculated by molecular docking (black bar) and MMGBSA (gray bar) for each compound. MMGBSA calculations were performed from selected structures every 10 ns over the last 100 ns of the simulation. For each ligand, its chemical structure is highlighted, and for the controls, the IC50 is reported in the literature. * IC50 obtained from [<a href="#B17-pharmaceuticals-15-00986" class="html-bibr">17</a>]. Full article ">

15 pages, 1898 KiB

Open AccessArticle

Optimization of 2-Aminoquinazolin-4-(3H)-one Derivatives as Potent Inhibitors of SARS-CoV-2: Improved Synthesis and Pharmacokinetic Properties

by Young Sup Shin, Jun Young Lee, Sangeun Jeon, Jung-Eun Cho, Subeen Myung, Min Seong Jang, Seungtaek Kim, Jong Hwan Song, Hyoung Rae Kim, Hyeung-geun Park, Lak Shin Jeong and Chul Min Park

Pharmaceuticals 2022, 15(7), 831; https://doi.org/10.3390/ph15070831 - 4 Jul 2022

Cited by 5 | Viewed by 2756

Abstract

We previously reported the potent antiviral effect of the 2-aminoquinazolin-4-(3H)-one 1, which shows significant activity (IC₅₀ = 0.23 μM) against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) with no cytotoxicity. However, it is necessary to improve the in vivo [...] Read more.

We previously reported the potent antiviral effect of the 2-aminoquinazolin-4-(3H)-one 1, which shows significant activity (IC₅₀ = 0.23 μM) against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) with no cytotoxicity. However, it is necessary to improve the in vivo pharmacokinetics of compound 1 because its area under the curve (AUC) and maximum plasma concentration are low. Here, we designed and synthesized N-substituted quinazolinone derivatives that had good pharmacokinetics and that retained their inhibitory activity against SARS-CoV-2. These compounds were conveniently prepared on a large scale through a one-pot reaction using Dimroth rearrangement as a key step. The synthesized compounds showed potent inhibitory activity, low binding to hERG channels, and good microsomal stability. In vivo pharmacokinetic studies showed that compound 2b had the highest exposure (AUC_24h = 41.57 μg∙h/mL) of the synthesized compounds. An in vivo single-dose toxicity evaluation of compound 2b at 250 and 500 mg/kg in rats resulted in no deaths and an approximate lethal dose greater than 500 mg/kg. This study shows that N-acetyl 2-aminoquinazolin-4-(3H)-one 2b is a promising lead compound for developing anti-SARS-CoV-2 agents. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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17 pages, 31961 KiB

Open AccessArticle

The Potential Complementary Role of Using Chinese Herbal Medicine with Western Medicine in Treating COVID-19 Patients: Pharmacology Network Analysis

by Yi-Chin Lu, Liang-Wei Tseng, Yu-Chieh Huang, Ching-Wei Yang, Yu-Chun Chen and Hsing-Yu Chen

Pharmaceuticals 2022, 15(7), 794; https://doi.org/10.3390/ph15070794 - 26 Jun 2022

Cited by 8 | Viewed by 3715

Abstract

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused a global pandemic in 2019—coronavirus disease (COVID-19). More and more Western medicine (WM) and Chinese herbal medicine (CHM) treatments have been used to treat COVID-19 patients, especially among Asian populations. However, the interactions between [...] Read more.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused a global pandemic in 2019—coronavirus disease (COVID-19). More and more Western medicine (WM) and Chinese herbal medicine (CHM) treatments have been used to treat COVID-19 patients, especially among Asian populations. However, the interactions between WM and CHM have not been studied. This study aims at using the network pharmacology approach to explore the potential complementary effects among commonly used CHM and WM in a clinical setting from a biomolecular perspective. Three well-published and widely used CHM formulas (National Research Institute of Chinese Medicine 101 (NRICM101), Qing-Fei-Pai-Du-Tang (QFPDT), Hua-Shi-Bai-Du-Formula (HSBDF)) and six categories of WM (Dexamethasone, Janus kinase inhibitors (JAKi), Anti-Interleukin-6 (Anti-IL6), anticoagulants, non-vitamin K antagonist oral anticoagulants (NOAC), and Aspirin) were included in the network pharmacology analysis. The target proteins on which these CHM and WM had direct effects were acquired from the STITCH database, and the potential molecular pathways were found in the REACTOME database. The COVID-19-related target proteins were obtained from the TTD database. For the three CHM formulas, QFPDT covered the most proteins (714), and 27 of them were COVID-19-related, while HSBDF and NRICM101 covered 624 (24 COVID-19-related) and 568 (25 COVID-19-related) proteins, respectively. On the other hand, WM covered COVID-19-related proteins more precisely and seemed different from CHM. The network pharmacology showed CHM formulas affected several inflammation-related proteins for COVID-19, including IL-10, TNF-α, IL-6, TLR3, and IL-8, in which Dexamethasone and Aspirin covered only IL-10 and TNF-α. JAK and IL-6 receptors were only inhibited by WM. The molecular pathways covered by CHM and WM also seemed mutually exclusive. WM had advantages in cytokine signaling, while CHM had an add-on effect on innate and adaptive immunity, including neutrophil regulation. WM and CHM could be used together to strengthen the anti-inflammation effects for COVID-19 from different pathways, and the combination of WM and CHM may achieve more promising results. These findings warrant further clinical studies about CHM and WM use for COVID-19 and other diseases. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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12 pages, 2902 KiB

Open AccessArticle

Analgesics Induce Alterations in the Expression of SARS-CoV-2 Entry and Arachidonic-Acid-Metabolizing Genes in the Mouse Lungs

by Fatima Khirfan, Yazun Jarrar, Tariq Al-Qirim, Khang Wen Goh, Qais Jarrar, Chrismawan Ardianto, Mohammad Awad, Hamzeh J. Al-Ameer, Wajdy Al-Awaida, Said Moshawih and Long Chiau Ming

Pharmaceuticals 2022, 15(6), 696; https://doi.org/10.3390/ph15060696 - 1 Jun 2022

Cited by 7 | Viewed by 3236

Abstract

Paracetamol and nonsteroidal anti-inflammatory drugs are widely used in the management of respiratory viral infections. This study aimed to determine the effects of the most commonly used analgesics (paracetamol, ibuprofen, and diclofenac) on the mRNA expression of severe acute respiratory syndrome coronavirus 2 [...] Read more.

Paracetamol and nonsteroidal anti-inflammatory drugs are widely used in the management of respiratory viral infections. This study aimed to determine the effects of the most commonly used analgesics (paracetamol, ibuprofen, and diclofenac) on the mRNA expression of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) entry and arachidonic-acid-metabolizing genes in mouse lungs. A total of twenty eight Balb/c mice were divided into four groups and treated separately with vehicle, paracetamol, ibuprofen, and diclofenac in clinically equivalent doses for 14 days. Then, the expressions of SARS-CoV-2 entry, ACE2, TMPRSS2, and Ctsl genes, in addition to the arachidonic-acid-metabolizing cyp450, cox, and alox genes, were analyzed using real-time PCR. Paracetamol increased the expressions of TMPRSS2 and Ctsl genes by 8.5 and 5.6 folds, respectively, while ibuprofen and diclofenac significantly decreased the expression of the ACE2 gene by more than 2.5 folds. In addition, all tested drugs downregulated (p < 0.05) cox2 gene expression, and paracetamol reduced the mRNA levels of cyp4a12 and 2j5. These molecular alterations in diclofenac and ibuprofen were associated with pathohistological alterations, where both analgesics induced the infiltration of inflammatory cells and airway wall thickening. It is concluded that analgesics such as paracetamol, ibuprofen, and diclofenac alter the expression of SARS-CoV-2 entry and arachidonic-acid-metabolizing genes in mouse lungs. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Figure 1
The changes in total body weight of the experimental mice. There was no significant change (p > 0.05, two-way ANOVA) in the total body weight after 14 days of analgesic treatment. Full article ">Figure 2
Histopathologic lungs analysis of animals after treatment. (A) Control lung section shows the normal structure of the bronchiole and adjacent alveoli. (B) Ibuprofen-treated mice representative lung section showing normal lung histology. (C) Diclofenac-treated mice lung tissue section showing normal bronchus and adjacent alveoli. (D) Paracetamol-treated mice lung tissue showing normal bronchial and alveolar tissues. Thick arrows indicate thickening in the alveolar wall; thin arrows indicate inflammatory cell infiltration. Tissue sections were stained with hematoxylin and eosin (scale bar 100 µm) and photographed at 40× magnification. Full article ">Figure 3
Relative mRNA expression of SARS-CoV-2 entry genes in the mouse lungs. * indicates a statistical alteration (p <0.05, one-way ANOVA test). Full article ">Figure 4
Expressions of SARS-CoV-2 entry genes ACE2 (A), TMPRSS2 (B), and Ctsl (C) in the lungs of NSAID- and paracetamol-treated mice. The mRNA expression of the targeted genes was quantified relative to Actin expression. Fold change indicates the ratio of mean expression of the NSAID- and paracetamol-treated to the control value. Negative values indicate a reduction in fold change. * indicates a statistical difference (p < 0.05, one-way ANOVA test) in comparison with the control group. Full article ">Figure 5
Expression of cox1 (A) and cox2 (B) genes in the lungs of NSAID- and paracetamol-treated mice. The target expression was quantified relative to the expression of Actin gene. Fold change is the ratio of mean expression of the NSAID- and paracetamol-treated to the control value. Negative values indicate a reduction in fold change. * indicates a statistical difference (p < 0.05, one-way ANOVA test) in comparison with the control group, while # indicates a statistical difference in comparison of diclofenac with other analgesics. Full article ">Figure 6
Expression of alox12 (A) and 15 (B) genes in the lungs of NSAID- and paracetamol-treated mice. The target expression was quantified relative to the expression of actin gene. Fold change is the ratio of mean expression of the NSAID- and paracetamol-treated to the control value. Negative values indicate a reduction in fold change; * indicates a statistical difference (p < 0.05, one-way ANOVA test) in comparison with the control group. Full article ">Figure 7
Expressions of cyp4a12 (A), cyp2j5 (B), and cyp2c29 (C) genes in the lungs of NSAID- and paracetamol-treated mice. The target expression was quantified relative to the expression of the actin gene. Fold change is the ratio of mean expression of the NSAID- and paracetamol-treated to the control value; negative values indicate a reduction in fold change; * indicates a statistical difference (p < 0.05, one-way ANOVA test) in comparison with the control group. Full article ">

16 pages, 2126 KiB

Open AccessArticle

Accurate Mass Identification of an Interfering Water Adduct and Strategies in Development and Validation of an LC-MS/MS Method for Quantification of MPI8, a Potent SARS-CoV-2 Main Protease Inhibitor, in Rat Plasma in Pharmacokinetic Studies

by Yang Wang, Huan Xie, Yugendar R. Alugubelli, Yuying Ma, Shiqing Xu, Jing Ma, Wenshe R. Liu and Dong Liang

Pharmaceuticals 2022, 15(6), 676; https://doi.org/10.3390/ph15060676 - 27 May 2022

Cited by 1 | Viewed by 2895

Abstract

MPI8, a peptidyl aldehyde, is a potent antiviral agent against coronavirus. Due to unique tri-peptide bonds and the formyl functional group, the bioassay of MPI8 in plasma was challenged by a strong interference from water MPI8. Using QTOF LC-MS/MS, we identified MPI8•H₂ [...] Read more.

MPI8, a peptidyl aldehyde, is a potent antiviral agent against coronavirus. Due to unique tri-peptide bonds and the formyl functional group, the bioassay of MPI8 in plasma was challenged by a strong interference from water MPI8. Using QTOF LC-MS/MS, we identified MPI8•H₂O as the major interference form that co-existed with MPI8 in aqueous and biological media. To avoid the resolution of MPI8 and MPI8•H₂O observed on reverse phase columns, we found that a Kinetex hydrophilic interaction liquid chromatography (HILIC) column provided co-elution of both MPI8 and MPI8•H₂O with a good single chromatographic peak and column retention of MPI8 which is suitable for quantification. Thus, a sensitive, specific, and reproducible LC-MS/MS method for the quantification of MPI8 in rat plasma was developed and validated using a triple QUAD LC-MS/MS. The chromatographic separation was achieved on a Kinetex HILIC column with a flow rate of 0.4 mL/min under gradient elution. The calibration curves were linear (r² > 0.99) over MPI8 concentrations from 0.5–500 ng/mL. The accuracy and precision are within acceptable guidance levels. The mean matrix effect and recovery were 139% and 73%, respectively. No significant degradation of MPI8 occurred under the experimental conditions. The method was successfully applied to a pharmacokinetic study of MPI8 after administration of MPI8 sulfonate in rats. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Full article ">Figure 1
Chemical structures of MPI8, its prodrug MPI8 sulfonate, and antipyrine (AP, internal standard). Full article ">Figure 2
MRM chromatograms for MPI8 (transitions m/z 601.4→545.3 and 601.4→157.2): Not-fully-separated multiple peaks on a Kinetex F5 column (A) and an Acquity HSS-T3 column (B); distinctly separated double peak chromatograms of MPI8 on a Synergi Fusion-RP column (C); single peak with good peak shape on a Kinetex HILIC column (D). The time program of the gradient: (A,B,D) Phase B was initially kept at 5% for 0.5 min, increased from 5% to 90% in the next 2.5 min, then decreased to 5% in 1.0 min, and kept stably at 5% for 1 min; (C) Phase B was initially at 20% for 0.5 min, increased from 20% to 90% in 4.5 min, kept at 90% for 2 min, decreased to initial concentration (20%) in 1 min and equilibrated for 2 min. Full article ">Figure 3
HR MS and MS/MS spectra of MPI8 and MPI8•H2O using an extracted sample from MPI8 spiked plasma at retention time of 4.5 min after column elution on an X500B QTOF mass spectrometer; (A1,A2) and (B1,B2) in positive mode, (C1,C2) and (D1,D2) in negative mode; the proposed fragmentation of product ions are shown on MS/MS spectra: (A1) protonated MPI8 [MPI8 + H]+ MS (m/z 601.3594), (A2) protonated MPI8 [MPI8 + H]+ MS/MS; (B1) [MPI8•H2O + Na]+ MS (m/z 641.3502) (B2) [MPI8•H2O + Na]+ MS/MS; (C1) deprotonated MPI8 [MPI8 −H]− MS (m/z 599.3433), (C2) deprotonated MPI8 [MPI8 − H]− MS/MS; (D1) [MPI8•H2O + Cl]− MS (m/z 653.3325), (D2) [MPI8•H2O + Cl]− MS/MS. Full article ">Figure 4
HR-MS extracted ion chromatograms of MPI8 and MPI8•H2O eluted on a Synergi Fusion-RP column; the peak at 5.2 min was MPI8; the peak at 4.5 min was MPI8•H2O, and the MPI8 signal was due to the in-source fragmentation: (A) MPI8 ([MPI8 + H]+) m/z 601.3596 in pink and MPI8•H2O ([MPI8•H2O + Na]+) m/z 641.3521 in blue in positive mode (B) MPI8 ([MPI8 −H]−) m/z 599.3450 in pink and MPI8•H2O ([MPI8•H2O + Cl]−) m/z 653.3323 in blue in negative mode. Full article ">Figure 5
The peak height and retention time of MPI8 (100 ng/mL) on a Kinetex HILIC column in isocratic elution at different concentrations of the mobile phase component of acetonitrile, from 5% to 90%. When the acetonitrile composition in the mobile phase was less than 10%, MPI8 was completely retained on the column, and the retention time mark was 5 min, which was the total running time of the test. Full article ">Figure 6
Representative MRM chromatograms: (A1) MPI8 in double blank rat plasma, (A2) IS in double blank plasma; (B1) MPI8 (0.5 ng/mL) spiked in blank rat plasma, (B2) IS (5 ng/mL) spiked in blank rat plasma; (C1) MPI8 in a rat plasma sample at 30 min after IV administration of MPI8 sulfonate at a single dose of 5 mg/kg, (C2) IS (5ng/mL) in a rat plasma sample at 30 min after IV administration of MPI8 sulfonate at a single dose of 5 mg/kg. There was no interference at the retention times of the analyte and IS, and no carryover for both IS (≤5% of average response) and MPI8 (≤20% of LLOQ). Full article ">Figure 7
Mean plasma concentration vs. time profiles of MPI8 in rat plasma after i.v. (n = 3) and oral (n = 3) administration (normal scale plots in normal graph and semi-log plots in inset graph). Twelve time-point plasma samples from rats dosed intravenously and 13 time-point plasma samples from rats dosed orally were collected. Three replicates for each time point were analyzed and reported as mean ± SD for each time-point. Full article ">

13 pages, 4656 KiB

Open AccessArticle

Discovery of Natural Lead Compound from Dendrobium sp. against SARS-CoV-2 Infection

by Jutamas Jiaranaikulwanitch, Wipawadee Yooin, Nopporn Chutiwitoonchai, Worathat Thitikornpong, Boonchoo Sritularak, Pornchai Rojsitthisak and Opa Vajragupta

Pharmaceuticals 2022, 15(5), 620; https://doi.org/10.3390/ph15050620 - 18 May 2022

Cited by 5 | Viewed by 2649

Abstract

Since the pandemic of severe acute respiratory syndrome coronavirus (SARS-CoV-2) in December 2019, the infection cases have quickly increased by more than 511 million people. The long epidemic outbreak over 28 months has affected health and economies worldwide. An alternative medicine appears to [...] Read more.

Since the pandemic of severe acute respiratory syndrome coronavirus (SARS-CoV-2) in December 2019, the infection cases have quickly increased by more than 511 million people. The long epidemic outbreak over 28 months has affected health and economies worldwide. An alternative medicine appears to be one choice to alleviate symptoms and reduce mortality during drug shortages. Dendrobium extract is one of the traditional medicines used for COVID-19 infection. Several compounds in Dendrobium sp. had been reported to exert pharmacological activities to treat common COVID-19-related symptoms. Herein, in silico screening of 83 compounds from Dendrobium sp. by using the SARS-CoV-2 spike protein receptor-binding domain (RBD) as a drug target was performed in searching for a new lead compound against SARS-CoV-2 infection. Four hit compounds showing good binding affinity were evaluated for antiviral infection activity. The new lead compound DB36, 5-methoxy-7-hydroxy-9,10-dihydro-1,4-phenanthrenequinone, was identified with the IC₅₀ value of 6.87 ± 3.07 µM. The binding mode revealed that DB36 bound with the spike protein at the host receptor, angiotensin-converting enzyme 2 (ACE2) binding motif, resulted in antiviral activity. This study substantiated the use of Dendrobium extract for the treatment of SARS-CoV-2 infection and has identified new potential chemical scaffolds for further drug development of SARS-CoV-2 entry inhibitors. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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20 pages, 4569 KiB

Open AccessArticle

Design of D-Amino Acids SARS-CoV-2 Main Protease Inhibitors Using the Cationic Peptide from Rattlesnake Venom as a Scaffold

by Raphael J. Eberle, Ian Gering, Markus Tusche, Philipp N. Ostermann, Lisa Müller, Ortwin Adams, Heiner Schaal, Danilo S. Olivier, Marcos S. Amaral, Raghuvir K. Arni, Dieter Willbold and Mônika A. Coronado

Pharmaceuticals 2022, 15(5), 540; https://doi.org/10.3390/ph15050540 - 27 Apr 2022

Cited by 13 | Viewed by 3532

Abstract

The C30 endopeptidase (3C-like protease; 3CL^pro) is essential for the life cycle of SARS-CoV-2 (severe acute respiratory syndrome-coronavirus-2) since it plays a pivotal role in viral replication and transcription and, hence, is a promising drug target. Molecules isolated from animals, insects, [...] Read more.

The C30 endopeptidase (3C-like protease; 3CL^pro) is essential for the life cycle of SARS-CoV-2 (severe acute respiratory syndrome-coronavirus-2) since it plays a pivotal role in viral replication and transcription and, hence, is a promising drug target. Molecules isolated from animals, insects, plants, or microorganisms can serve as a scaffold for the design of novel biopharmaceutical products. Crotamine, a small cationic peptide from the venom of the rattlesnake Crotalus durissus terrificus, has been the focus of many studies since it exhibits activities such as analgesic, in vitro antibacterial, and hemolytic activities. The crotamine derivative L-peptides (L-CDP) that inhibit the 3CL protease in the low µM range were examined since they are susceptible to proteolytic degradation; we explored the utility of their D-enantiomers form. Comparative uptake inhibition analysis showed D-CDP as a promising prototype for a D-peptide-based drug. We also found that the D-peptides can impair SARS-CoV-2 replication in vivo, probably targeting the viral protease 3CL^pro. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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25 pages, 2081 KiB

Open AccessArticle

Hypericum perforatum and Its Ingredients Hypericin and Pseudohypericin Demonstrate an Antiviral Activity against SARS-CoV-2

by Fakry F. Mohamed, Darisuren Anhlan, Michael Schöfbänker, André Schreiber, Nica Classen, Andreas Hensel, Georg Hempel, Wolfgang Scholz, Joachim Kühn, Eike R. Hrincius and Stephan Ludwig

Pharmaceuticals 2022, 15(5), 530; https://doi.org/10.3390/ph15050530 - 25 Apr 2022

Cited by 30 | Viewed by 8730

Abstract

For almost two years, the COVID-19 pandemic has constituted a major challenge to human health, particularly due to the lack of efficient antivirals to be used against the virus during routine treatment interventions. Multiple treatment options have been investigated for their potential inhibitory [...] Read more.

For almost two years, the COVID-19 pandemic has constituted a major challenge to human health, particularly due to the lack of efficient antivirals to be used against the virus during routine treatment interventions. Multiple treatment options have been investigated for their potential inhibitory effect on SARS-CoV-2. Natural products, such as plant extracts, may be a promising option, as they have shown an antiviral activity against other viruses in the past. Here, a quantified extract of Hypericum perforatum was tested and found to possess a potent antiviral activity against SARS-CoV-2. The antiviral potency of the extract could be attributed to the naphtodianthrones hypericin and pseudohypericin, in contrast to other tested ingredients of the plant material, which did not show any antiviral activity. Hypericum perforatum and its main active ingredient hypericin were also effective against different SARS-CoV-2 variants (Alpha, Beta, Delta, and Omicron). Concerning its mechanism of action, evidence was obtained that Hypericum perforatum and hypericin may hold a direct virus-blocking effect against SARS-CoV-2 virus particles. Taken together, the presented data clearly emphasize the promising antiviral activity of Hypericum perforatum and its active ingredients against SARS-CoV-2 infections. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Graphical abstract

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The antiviral activity of Hypericum perforatum (HP1) against the pseudo-typed VSV virus carrying the SARS-CoV-2 S protein. (A,C) Vero cells were seeded overnight, and on the next day, cells and the VSV-pseudo-typed virus were incubated with Hypericum perforatum (HP1) or solvent control (DMSO) for 1 h prior to infection, at 37 °C or room temperature, respectively. After pre-incubation, infection was performed with a MOI of 0.01 for 1 h, and cells were finally washed and incubated without further treatments. After 16–18 h, GFP signal was visualized under fluorescent microscope. (A) GFP-positive cells as % of control are shown (mean and s.d.), and one-way ANOVA with Dunnett’s multiple comparisons was done by comparing each value with the control. (C) Dose–response curve of the normalized GFP-positive cell values as % of control is depicted (mean and s.d.). (B,D) Vero cells were seeded overnight, and on the next day, incubation with HP1 or solvent control was initiated. 24 h after the start of incubation, the MTT assay-based cytotoxicity was measured. (B) Cell viability as % of control is shown (mean and s.d.), and one-way ANOVA with Dunnett’s multiple comparisons was done by comparing each value with the control. (D) Dose–response curve of the normalized cytotoxicity values as % of control is depicted (mean and s.d.). * for p ≤ 0.05, ** for p ≤ 0.01, and **** for p ≤ 0.0001. Full article ">Figure 2
Hypericin and pseudohypericin are key components in the Hypericum perforatum (HP1) extract that are antivirally effective against the pseudo-typed VSV virus. (A–E) Vero cells were seeded overnight, and the next day, different concentrations of hypericin (HY), pseudohypericin (PS), hyperforin (HF), procyanidin-C1 (PRO), and quercetin-3-O-glucuronid (ingredients of HP1 extract) were applied onto the cells for indicated time points, as solvent-treated cells (DMSO) served as control. In addition, Staurosporine-treated cells served as positive control. After the incubations, the MTT assay-based cytotoxicity was measured, cell viability as % of solvent control is shown (mean and s.d), and two-way ANOVA with Dunnett’s Multiple comparisons was done by comparing each value with the solvent control at each time point. (F–J) Vero cells were seeded overnight, and on the next day, cells and the VSV-pseudo-typed virus were incubated with the indicated substances or solvent control (DMSO) for 1 h prior to infection, at 37 °C or room temperature, respectively. After the pre-incubation, infection was performed with a MOI of 0.01 for 1 h, and cells were finally washed and incubated without further treatments. GFP-positive cells as % of solvent control are shown (mean and s.d.), and Student’s t-test with Welch’s correction was applied (n.d. means non-detected, while n.s. means non-significant statistical difference). * for p ≤ 0.05, ** for p ≤ 0.01, *** for p ≤ 0.001, and **** for p ≤ 0.0001. Full article ">Figure 3
Hypericin and pseudohypericin showed a strong antiviral activity against the pseudo-typed VSV virus. (A,C,E,G) Vero cells were seeded overnight, and the next day, prior to infection (MOI = 0.01), both cells and the VSV pseudo-typed virus were incubated with the indicated substances or solvent control (DMSO) for 1 h, at 37 °C and room temperature, respectively. After the pre-incubation, cells were infected for 1h and finally washed and incubated without further treatments. (A,E) GFP-positive cells as % of solvent control are shown (mean and s.d.), and one-way ANOVA with Dunnett’s multiple comparisons was done by comparing each value with the control. (B,D,F,H) Vero cells were seeded overnight, and the next day, incubation with the indicated substances or solvent control (DMSO) was initiated. After the 24 h incubation, MTT assay-based cytotoxicity was measured. (B,F) Cell viability as % of solvent control is shown (mean and s.d.), and one-way ANOVA with Dunnett’s multiple comparisons was done by comparing each value with the solvent control. (C,G) Dose–response curve of the normalized GFP-positive cell values as % of solvent control is depicted (mean and s.d.). (D,H) Dose–response curve of the normalized cytotoxicity values as % of solvent control is depicted (mean and s.d.). n.d means non-detected, * for p ≤ 0.05, *** for p ≤ 0.001, and **** for p ≤ 0.0001. Full article ">Figure 4
Hypericum perforatum (HP1) acts as an antiviral against ancestral SARS-CoV-2. (A,C) Vero cells were seeded overnight, and on the next day, prior to infection (MOI = 0.05), cells were incubated at 37 °C for 1h with infection-DMEM containing either solvent control (DMSO) or HP1. Concurrently, SARS-CoV-2 was incubated for 1 h at room temperature in infection-PBS that contained either DMSO or HP1. After infection (37 °C/1 h), cells were further incubated in infection-DMEM including either DMSO or HP1. After 24 h, virus supernatants were collected and subjected to plaque assay. (A) Results are expressed as PFU/mL (mean and s.d.), and one-way ANOVA with Dunnett’s multiple comparisons was done by comparing each value with the control. (C) Dose–response curve of the normalized virus titer values as % of solvent control is depicted (mean and s.d.). (B,D–F) Vero cells were seeded overnight, and on the next day, cells were incubated for 24 h with infection-DMEM that contained either solvent control (DMSO) or HP1. After incubation, the MTT assay-based cytotoxicity was measured. (B,E) Cell viability as % of solvent control is shown (mean and s.d.), and one-way ANOVA with Dunnett’s multiple comparisons was done by comparing each value with the control. (D,F) Dose–response curve of the normalized cytotoxicity values as % of solvent control is depicted (mean and s.d.). * for p ≤ 0.05, ** for p ≤ 0.01, and **** for p ≤ 0.0001. Full article ">Figure 5
Hypericum perforatum (HP1) and hypericin (HY) inhibit the growth of different SARS-CoV-2 variants. (A–F) Vero cells were seeded overnight, and the next day, before being infected, cells were incubated in infection-DMEM containing solvent control (DMSO) or either (A–C) HP1 or (D–F) hypericin for 1h, at 37 °C. Meanwhile, SARS-CoV-2 variants were also preincubated (1 h at room temperature) before infection in infection-PBS with either solvent control or (A–C) HP1 or (D–F) hypericin. After pre-incubation, virus infection was performed at a MOI of 0.05 for 1 h. After infection, cells were either incubated with solvent control or (A–C) HP1 or (D–F) hypericin. After 24 h infection, virus supernatants were collected, and virus titration was done by plaque assays. (A–F) Obtained data are shown as PFU/mL (mean and s.d.). Full article ">Figure 6
Hypericum perforatum (HP1) and hypericin (HY) carry direct SARS-CoV-2 virus-blocking activities. (A–C) Vero cells were seeded overnight. The next day, different treatment protocols with HP1 or hypericin were applied. The treatment conditions included (i) only pre-treatment of cells (for 1 h at 37 °C) in infection-DMEM containing solvent control (DMSO) or HP1 or hypericin, (ii) only pre-treatment of SARS-CoV-2 (for 1 h at room temperature) in infection-PBS containing solvent control or HP1 or hypericin, or (iii) only post-treatment of cells after infection in infection-DMEM (at 37 °C) containing solvent control or HP1 or hypericin. As control, the combined treatment protocol of pre-treatment of cells and SARS-CoV-2 and post-treatment of cells was included as well (Full treatment). The SARS-CoV-2 infection was conducted at MOI of 0.05 or 1, as the total length of the infection experiment was (A,B) 24 h or (C) 8 h, respectively. (A–C) After the depicted length of experiments, virus supernatants were harvested, virus titration was done with plaque assay, results are shown as PFU/mL (means and s.d.), and two-way ANOVA with Sidak’s multiple comparisons was done by comparing each value to its respective solvent control. (D) SARS-CoV-2 was incubated for 1 h at room temperature with solvent control (DMSO) or HP1 or hypericin in an infection-PBS mix and directly submitted to plaque assay. Obtained data are expressed as PFU/mL (mean and s.d.), and one-way ANOVA with Dunnett’s multiple comparisons was done by comparing each value to the solvent control. (E) After Vero cells were seeded on cover slips overnight, cells were infected with 1 h pre-treated (either with DMSO or 15 µg/mL HP1 or 100 ng/mL hypericin (HY)) SARS-CoV-2 virus. Mock-infected cells served as control. Then, 2, 4, 6, and 8 h after infection, cold methanol (–20 °c) was used for cell fixation, and indirect immunofluorescence staining of the SARS-CoV-2 nucleoprotein (green) and nuclei (blue) was conducted. Exposure times for each channel where fixed on the 8 h infected and DMSO-treated samples (scale bar represents 50µM). (F) The day after seeding, Vero cells were infected with SARS-CoV-2 (MOI = 0.05) for 1 h. After infection, cells were either incubated with solvent control or hypericin (250 ng/mL) in infection-DMEM. Supernatants were collected 24 h after infection and submitted to plaque assay. Obtained data are shown as PFU/mL, and Student’s t-test with Welch’s corrections was done. n.d means non-detected. * for p ≤ 0.05, ** for p ≤ 0.01, and **** for p ≤ 0.0001. Full article ">Figure 7
Hypericum perforatum (HP1) and its ingredient hypericin (HY) showed an antiviral capacity against the pseudotyped VSV virus carrying SARS-CoV-2 S protein of the Omicron variant. Vero cells were seeded overnight, and the day after, cells were treated for 1 h at 37 °C with fresh DMEM-10% FCS containing the solvent control (DMSO) or the indicated concentration of HP1 or hypericin. In parallel, the pseudo-typed virus carrying (A) the genuine SARS-CoV-2 S protein (Wuhan S protein sequence) or (B) the Omicron variant S protein (Omicron S protein sequence) was incubated with either solvent control or the indicated concentration of each substance at room temperature for 1 h. After the 1 h incubation, the virus solution (MOI = 0.01) was applied on cells for 1 h, at 37 °C for infection, followed by a DMEM-10% FCS wash step, and a final application of fresh DMEM-10 % FCS. On the next day, GFP-positive cells were counted by Celigo Image Cytometer (Nexcelom Bioscience, Lawrence, MA, USA). GFP-positive cells as % of control are shown (mean and s.d.), and one-way ANOVA with Dunnett’s multiple comparisons was done by comparing each value with the control. **** for p ≤ 0.0001. Full article ">

22 pages, 5112 KiB

Open AccessArticle

Use of Early Donated COVID-19 Convalescent Plasma Is Optimal to Preserve the Integrity of Lymphatic Endothelial Cells

by Nada Amri, Rémi Bégin, Nolwenn Tessier, Laurent Vachon, Louis Villeneuve, Philippe Bégin, Renée Bazin, Lionel Loubaki and Catherine Martel

Pharmaceuticals 2022, 15(3), 365; https://doi.org/10.3390/ph15030365 - 17 Mar 2022

Cited by 5 | Viewed by 3516

Abstract

Convalescent plasma therapy (CPT) has gained significant attention since the onset of the coronavirus disease 2019 (COVID-19) pandemic. However, clinical trials designed to study the efficacy of CPT based on antibody concentrations were inconclusive. Lymphatic transport is at the interplay between the immune [...] Read more.

Convalescent plasma therapy (CPT) has gained significant attention since the onset of the coronavirus disease 2019 (COVID-19) pandemic. However, clinical trials designed to study the efficacy of CPT based on antibody concentrations were inconclusive. Lymphatic transport is at the interplay between the immune response and the resolution of inflammation from peripheral tissues, including the artery wall. As vascular complications are a key pathogenic mechanism in COVID-19, leading to inflammation and multiple organ failure, we believe that sustaining lymphatic vessel function should be considered to define optimal CPT. We herein sought to determine what specific COVID-19 convalescent plasma (CCP) characteristics should be considered to limit inflammation-driven lymphatic endothelial cells (LEC) dysfunction. CCP donated 16 to 100 days after the last day of symptoms was characterized and incubated on inflammation-elicited adult human dermal LEC (aHDLEC). Plasma analysis revealed that late donation correlates with higher concentration of circulating pro-inflammatory cytokines. Conversely, extracellular vesicles (EVs) derived from LEC are more abundant in early donated plasma (r = −0.413, p = 0.004). Thus, secretion of LEC-EVs by an impaired endothelium could be an alarm signal that instigate the self-defense of peripheral lymphatic vessels against an excessive inflammation. Indeed, in vitro experiments suggest that CCP obtained rapidly following the onset of symptoms does not damage the aHDLEC junctions as much as late-donated plasma. We identified a particular signature of CCP that would counteract the effects of an excessive inflammation on the lymphatic endothelium. Accordingly, an easy and efficient selection of convalescent plasma based on time of donation would be essential to promote the preservation of the lymphatic and immune system of infected patients. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Graphical abstract

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Elevated antibody concentrations and prolonged symptoms are detrimental for the lymphatic endothelium integrity. (A) Correlation between the duration of symptoms (square-root transformation) and the concentration of plasmatic SARS-CoV-2- receptor-binding-domain antibodies measured by ELISA. (B) Correlation between the duration of symptoms and MitoSOXTM Red-negative cells measured by flow cytometry after incubating aHDLEC with convalescent plasma for 4 h and cytokines cocktail for 20 h. (C) Correlation between the concentration of plasmatic RBD antibodies measured by ELISA and the permeability of the endothelium measured by spectrophotometry (absorbance of OVA-488) after incubating human LEC with convalescent plasma for 4 h and cytokines cocktail for 20 h. Each point represents a treatment. Significance (p < 0.05) was determined by a Pearson correlation. A square-root transformation of the duration of symptoms to reach normal distribution was performed. RBD—receptor-binding domain; O.D—optical density; OVA—ovalbumin; aHDLEC—adult human dermal lymphatic endothelial cells. Full article ">Figure 2
The severity of symptoms correlates with the duration of symptoms, age, receptor-binding-domain antibodies of donors and cell viability of lymphatic endothelial cells. (A) Severity was quantified using a questionnaire answered by donors and graded as follows: asymptomatic, mild, moderate and severe. Severe symptoms were correlated with a higher duration of symptoms (square-root transformation to reach a normal distribution). (B) Correlations between severity of the symptoms and the age of the donors were performed. (C) Severity was correlated with the concentration of antibodies measured by ELISA. (D,E) Treated aHDLEC were labeled with Annexin V and PI and analyzed by flow cytometry. The severity of the symptoms was correlated with cells in late apoptosis/necrosis (D) and in early apoptosis (E) represented by Annexin V- and PI-positive cells and Annexin V-positive and PI-negative cells, respectively. Cells are given as percentages relative to cells treated with control plasma. Significance was determined by a Spearman correlation. p < 0.05 was considered significant. RBD, receptor-binding domain; O.D, optical density; PI, propidium iodide; aHDLEC, adult human dermal lymphatic endothelial cells. Full article ">Figure 3
Early donation of COVID-19 convalescent plasma is a good predictor of preserved endothelial integrity. (A,B) Treated aHDLEC (convalescent plasma for 4 h and cytokines for 20 h) were harvested and the mRNA expression, measured by RT-qPCR, of SELE (A) and ICAM1 (B) was correlated with the time of donation since the onset of symptoms. (C) Following treatment, cells were fixed in paraformaldehyde (PFA) 2% and incubated with anti-VE-Cadherin antibodies. The intensity of VE-Cadherin relative to cells treated with control plasma was correlated to the duration between the onset of symptoms and the donation. (D) Immunofluorescence images of treated aHDLEC incubated with CCP donated at 27 days (upper panels) and 101 days (lower panels) post onset of symptoms. The left panels show the expression of VE-Cadherin and DAPI, whereas the middle panel represents the WGA staining. The right panels show the representation of the merged staining. Scale bar = 50 μm. (E) VE-Cadherin mRNA expression was correlated to the duration between the onset of symptoms and donation (∆). (F) Endothelial permeability was analyzed by the relative concentration of ovalbumin-488 measured following migration through the endothelium compared to control. The concentration of OVA-488 was correlated with the duration between the onset of symptoms and the donation. Significance was determined by Pearson correlation and p < 0.05 was considered significant. ∆, time of donation since onset of symptoms; SELE, gene coding for E-selectin; ICAM1, gene coding for intercellular adhesion molecule 1; WGA, wheat germ agglutinin; CDH5, gene coding for VE-Cadherin; OVA, ovalbumin; aHDLEC, adult human dermal lymphatic endothelial cells; CCP, COVID-19 convalescent plasma. Full article ">Figure 4
Late donation correlates with higher concentration of circulating pro-inflammatory cytokines. (A–I). Concentration of pro-inflammatory cytokines and mediators contained in the convalescent plasma was analyzed using a Multiplex kit. The duration between onset of symptoms and donation (∆) was correlated to interleukin 1 beta (IL-1β) (A), IL-13 (B), IL-12 p70 (C), interferon gamma (IFNγ) (D), IL17A (E), IP-10 (F), MIP1 (G), IFNα (H) and IL-1α (I). Significance was determined by a Pearson correlation and p < 0.05 was considered significant. ∆, time of donation since onset of symptoms. IL, interleukin; IFN, interferon; IP10, interferon gamma-induced protein 10; MIP1, macrophage inflammatory protein 1. Full article ">Figure 5
Characterization of extracellular vesicles and correlation with duration between onset of symptoms and donation. (A–G) EVs gating strategy (side scatter) for CFSE+ EVs (A), MHCI+ EVs (B), CD45+ EVs (C), CD235+ EVs (D), CD45− podoplanin+ EVs (E), CD62e+ EVs (F) and CLEC2+ EVs (G). (H) Concentration of EVs in CCP measured by flow cytometry. Kruskal–Wallis test with a Dunn post hoc test was performed. (I) Concentration of CD45− podoplanin+ EVs in convalescent plasma and correlation with the duration between the onset of symptoms and the donation. Significance was determined by a Pearson correlation after a logarithmic transformation of CD45− podoplanin+ EVs concentration to reach a normal distribution. p < 0.05 was considered significant. **** p < 0.0001 significantly different from podoplanin+ EVs. #### p < 0.0001 significantly different from CD62e+ EVs. EVs, extracellular vesicles; CCP, COVID-19 convalescent plasma; CFSE, carboxyfluorescein succinimidyl ester; MHCI, major histocompatibility complex I; PDPN, podoplanin; CLEC2, C-type lectin-like type II; ∆, time of donation since onset of symptoms. Full article ">Figure 6
Secretion of lymphatic endothelial cells derived extracellular vesicles is a marker of impaired integrity. (A) Treated aHDLEC (convalescent plasma for 4 h and cytokines for 20 h) labeled by the MitoSOXTM Red probe and correlated with the quantity of EVs shed by the lymphatic endothelial cells. (B,C) Cells in sub-G0/G1 (B) and G2/M (C) phases of the cell cycle, determined by cell cycle analysis using PI, were correlated to the quantity of EVs secreted by the lymphatic endothelium. (D,E). Treated aHDLEC were harvested and expression of FLT4 was assessed by RT-qPCR (D) and immunoblot (E) followed by a normalization onto the expression of ACTB or α/ß tubulin, respectively. The measured expression of VEGFR-3 was then correlated with the quantity of EVs secreted by the endothelium. (F) Immunoblot analysis for VEGFR-3 and the loading control α/ß tubulin. The left panel represents aHDLEC secreting low quantities of CD45− podoplanin+ EVs (less than 120,000 EVs) following incubation with CCP, and the right represents aHDLEC secreting high quantities of CD45− podoplanin+ EVs (more than 380,000 EVs). Significance was determined by a Pearson correlation and p < 0.05 was considered significant. EVs, extracellular vesicles; PI, propidium iodide; CCP, COVID-19 convalescent plasma; VEGFR-3, vascular endothelial growth factor receptor 3; FLT4, gene coding for VEGFR-3; aHDLEC, adult human dermal lymphatic endothelial cells; AU, arbitrary unit. Full article ">

18 pages, 7756 KiB

Open AccessArticle

Efficacy of Adaptogens in Patients with Long COVID-19: A Randomized, Quadruple-Blind, Placebo-Controlled Trial

by Irina Karosanidze, Ushangi Kiladze, Nino Kirtadze, Mikhail Giorgadze, Nana Amashukeli, Nino Parulava, Neli Iluridze, Nana Kikabidze, Nana Gudavadze, Lali Gelashvili, Vazha Koberidze, Eka Gigashvili, Natela Jajanidze, Naira Latsabidze, Nato Mamageishvili, Ramaz Shengelia, Areg Hovhannisyan and Alexander Panossian

Pharmaceuticals 2022, 15(3), 345; https://doi.org/10.3390/ph15030345 - 11 Mar 2022

Cited by 19 | Viewed by 8255

Abstract

Currently, no effective treatment of comorbid complications or COVID-19 long-haulers during convalescence is known. This randomized, quadruple-blind, placebo-controlled trial aimed to assess the efficacy of adaptogens on the recovery of patients with Long COVID symptoms. One hundred patients with confirmed positive SARS-CoV-2 test, [...] Read more.

Currently, no effective treatment of comorbid complications or COVID-19 long-haulers during convalescence is known. This randomized, quadruple-blind, placebo-controlled trial aimed to assess the efficacy of adaptogens on the recovery of patients with Long COVID symptoms. One hundred patients with confirmed positive SARS-CoV-2 test, discharged from COVID Hotel isolation, Intensive Care Unit (ICU), or Online Clinics, and who experienced at least three of nine Long COVID symptoms (fatigue, headache, respiratory insufficiency, cognitive performance, mood disorders, loss of smell, taste, and hair, sweatiness, cough, pain in joints, muscles, and chest) in the 30 days before randomization were included in the study of the efficacy of Chisan^®/ADAPT-232 (a fixed combination of adaptogens Rhodiola, Eleutherococcus, and Schisandra) supplementation for two weeks. Chisan^® decreased the duration of fatigue and pain for one and two days, respectively, in 50% of patients. The number of patients with lack of fatigue and pain symptoms was significantly less in the Chisan^® treatment group than in the placebo group on Days 9 (39% vs. 57%, pain relief, p = 0.0019) and 11 (28% vs. 43%, relief of fatigue, * p = 0.0157). Significant relief of severity of all Long COVID symptoms over the time of treatment and the follow-up period was observed in both groups of patients, notably decreasing the level of anxiety and depression from mild and moderate to normal, as well as increasing cognitive performance in patients in the d2 test for attention and increasing their physical activity and workout (daily walk time). However, the significant difference between placebo and Chisan^® treatment was observed only with a workout (daily walk time) and relieving respiratory insufficiency (cough). A clinical assessment of blood markers of the inflammatory response (C-reactive protein) and blood coagulation (D-dimer) did not reveal any significant difference over time between treatment groups except significantly lower IL-6 in the Chisan^® treatment group. Furthermore, a significant difference between the placebo and Chisan^® treatment was observed for creatinine: Chisan^® significantly decreased blood creatinine compared to the placebo, suggesting prevention of renal failure progression in Long COVID. In this study, we, for the first time, demonstrate that adaptogens can increase physical performance in Long COVID and reduce the duration of fatigue and chronic pain. It also suggests that Chisan^®/ADAPT-232 might be useful for preventing the progression of renal failure associated with increasing creatinine. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Figure 1
(a) Workout time (mean ± SEM) of patients in group A (ADAPT-232) and group B (placebo) over the time from Day 1 to Day 21. The changes from the baseline within groups A and B over time were significant (p < 0.0001; calculated by repeated measures ANOVA); two-way ANOVA estimated the significant interaction between treatment groups over time; *—p = 0.0148. (b) Between-groups comparison of the changes of workout from the baseline over time shows significant interaction (p < 0.0001) and very significant difference (p < 0.0001) between groups A and B. The ADAPT-232 treatment significantly increases patients’ workouts compared to placebo. ***—p< 0.001. For details of statistical analysis, see <a href="#app1-pharmaceuticals-15-00345" class="html-app">Supplement 2</a>. Full article ">Figure 2
The changes from the baseline of the levels (mean ± SEM) of (a) creatinine and (b) C-reactive protein in the blood of patients in group A (ADAPT-232) and group B (placebo) over the time from Day 1 to Day 21. Between-groups comparison of the creatinine shows a significant difference (p = 0.0012) between groups A and B. The ADAPT-232 treatment significantly decreased blood creatinine compared to placebo. For details of statistical analysis, see <a href="#app1-pharmaceuticals-15-00345" class="html-app">Supplement 2</a>. Between-groups comparison of the changes of C-reactive protein in blood from the baseline over time shows no interaction (p = 0.7100) and no significant difference (p = 0.1276) between groups A and B. The ADAPT-232 treatment has no statistically significant effect on C-reactive protein level in blood compared to placebo. *—p < 0.05 and **—p< 0.01. For details of statistical analysis, see <a href="#app1-pharmaceuticals-15-00345" class="html-app">Supplement 2</a>. Full article ">Figure 3
The changes from the baseline of the levels (mean ± SEM) of (a) cytokine IL-6 and (b) D-dimer in the blood of patients in group A (ADAPT-232) and group B (placebo) over the time from Day 1 to Day 21. Between-groups comparison of the changes of cytokine IL-6 level in the blood from the baseline over time shows no interaction (p = 0.4369) and no significant difference (p = 0.5879) between groups A and B. The ADAPT-232 treatment has no statistically significant effect on cytokine IL-6 in blood compared to placebo. Between-groups comparison of the changes of blood D-dimer level from the baseline over time shows no interaction (p = 0.8920) and no significant difference (p = 0.5782) between groups A and B. The ADAPT-232 treatment has no statistically significant effect on blood D-dimer compared to placebo. *—p < 0.05. For details of statistical analysis, see <a href="#app1-pharmaceuticals-15-00345" class="html-app">Supplement 2</a>. Full article ">Figure 4
Kaplan–Meier curves show the decrease of the duration of fatigue and pain over the time from randomization (Day 1) to the end of the treatment (Day 14) and followed up for one week (Day 21) and the number of patients who experienced these symptoms of Long COVID: (a) fatigue, (b) pain. *—p < 0.05. Full article ">Figure 5
Schematic diagram of the trial: disposition of patients. Full article ">

16 pages, 2246 KiB

Open AccessArticle

Mineralocorticoid Receptor Antagonist (Potassium Canrenoate) Does Not Influence Outcome in the Treatment of COVID-19-Associated Pneumonia and Fibrosis—A Randomized Placebo Controlled Clinical Trial

by Katarzyna Kotfis, Igor Karolak, Kacper Lechowicz, Małgorzata Zegan-Barańska, Agnieszka Pikulska, Paulina Niedźwiedzka-Rystwej, Miłosz Kawa, Jerzy Sieńko, Aleksandra Szylińska and Magda Wiśniewska

Pharmaceuticals 2022, 15(2), 200; https://doi.org/10.3390/ph15020200 - 5 Feb 2022

Cited by 5 | Viewed by 2582

Abstract

In December 2019 the SARS-CoV-2 virus appeared in the world, mainly presenting as an acute infection of the lower respiratory tract, namely pneumonia. Nearly 10% of all patients show significant pulmonary fibrotic changes after the infection. The aim of this study was to [...] Read more.

In December 2019 the SARS-CoV-2 virus appeared in the world, mainly presenting as an acute infection of the lower respiratory tract, namely pneumonia. Nearly 10% of all patients show significant pulmonary fibrotic changes after the infection. The aim of this study was to evaluate the effectiveness and safety of potassium canrenoate in the treatment of COVID-19-associated pneumonia and pulmonary fibrosis. We performed a randomized clinical trial (RCT) of potassium canrenoate vs placebo. A total of 55 patients were randomized and 49 were included in the final analysis (24 allocated to the intervention group and 25 allocated to the control group). Patients were assessed by physical examination, lung ultrasound, CT imaging and blood samples that underwent biochemical analysis. This RCT has shown that the administration of potassium canrenoate to patients with COVID-19 induced pneumonia was not associated with shorter mechanical ventilation time, shorter passive oxygenation, shorter length of hospitalization or less fibrotic changes on CT imaging. The overall mortality rate was not significantly different between the two groups. Adverse events recorded in this study were not significantly increased by the administration of potassium canrenoate. The negative outcome of the study may be associated with the relatively small number of patients included. Any possible benefits from the use of potassium canrenoate as an antifibrotic drug in COVID-19 patients require further investigation. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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14 pages, 6398 KiB

Open AccessArticle

Novel Small-Molecule Inhibitors of the SARS-CoV-2 Spike Protein Binding to Neuropilin 1

by Anja Kolarič, Marko Jukič and Urban Bren

Pharmaceuticals 2022, 15(2), 165; https://doi.org/10.3390/ph15020165 - 28 Jan 2022

Cited by 22 | Viewed by 5064

Abstract

Furin cleavage of the SARS-CoV-2 spike protein results in a polybasic terminal sequence termed the C-end rule (CendR), which is responsible for the binding to neuropilin 1 (NRP1), enhancing viral infectivity and entry into the cell. Here we report the identification of 20 [...] Read more.

Furin cleavage of the SARS-CoV-2 spike protein results in a polybasic terminal sequence termed the C-end rule (CendR), which is responsible for the binding to neuropilin 1 (NRP1), enhancing viral infectivity and entry into the cell. Here we report the identification of 20 small-molecule inhibitors that emerged from a virtual screening of nearly 950,000 drug-like compounds that bind with high probability to the CendR-binding pocket of NRP1. In a spike NRP1 binding assay, two of these compounds displayed a stronger inhibition of spike protein binding to NRP1 than the known NRP1 antagonist EG00229, for which the inhibition of the CendR peptide binding to NRP1 was also experimentally confirmed. These compounds present a good starting point for the design of small-molecule antagonists against the SARS-CoV-2 viral entry. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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14 pages, 2457 KiB

Open AccessArticle

Atazanavir Is a Competitive Inhibitor of SARS-CoV-2 M^pro, Impairing Variants Replication In Vitro and In Vivo

by Otávio Augusto Chaves, Carolina Q. Sacramento, André C. Ferreira, Mayara Mattos, Natalia Fintelman-Rodrigues, Jairo R. Temerozo, Leonardo Vazquez, Douglas Pereira Pinto, Gabriel P. E. da Silveira, Laís Bastos da Fonseca, Heliana Martins Pereira, Aluana Santana Carlos, Joana C. d’Avila, João P. B. Viola, Robson Q. Monteiro, Patrícia T. Bozza, Hugo Caire Castro-Faria-Neto and Thiago Moreno L. Souza

Pharmaceuticals 2022, 15(1), 21; https://doi.org/10.3390/ph15010021 - 24 Dec 2021

Cited by 27 | Viewed by 4952

Abstract

Atazanavir (ATV) has already been considered as a potential repurposing drug to 2019 coronavirus disease (COVID-19); however, there are controversial reports on its mechanism of action and effectiveness as anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Through the pre-clinical chain of experiments: enzymatic, [...] Read more.

Atazanavir (ATV) has already been considered as a potential repurposing drug to 2019 coronavirus disease (COVID-19); however, there are controversial reports on its mechanism of action and effectiveness as anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Through the pre-clinical chain of experiments: enzymatic, molecular docking, cell-based and in vivo assays, it is demonstrated here that both SARS-CoV-2 B.1 lineage and variant of concern gamma are susceptible to this antiretroviral. Enzymatic assays and molecular docking calculations showed that SARS-CoV-2 main protease (M^pro) was inhibited by ATV, with Morrison’s inhibitory constant (K_i) 1.5-fold higher than GC376 (a positive control) dependent of the catalytic water (H₂O_cat) content. ATV was a competitive inhibitor, increasing the M^pro’s Michaelis–Menten (K_m) more than sixfold. Cell-based assays indicated that different lineages of SARS-CoV-2 is susceptible to ATV. Using oral administration of ATV in mice to reach plasmatic exposure similar to humans, transgenic mice expression in human angiotensin converting enzyme 2 (K18-hACE2) were partially protected against lethal challenge with SARS-CoV-2 gamma. Moreover, less cell death and inflammation were observed in the lung from infected and treated mice. Our studies may contribute to a better comprehension of the M^pro/ATV interaction, which could pave the way to the development of specific inhibitors of this viral protease. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Figure 1
The 2D- and 3D-chemical structure for atazanavir (ATV). Full article ">Figure 2
(A) The ATV and GC376 (positive control) activity on 88.8 nM Mpro velocity at 0–10 μM of inhibitor. (B) Michaelis–Menten plot for 88.8 nM Mpro incubated with substrate concentrations from 0 to 100 μM in the presence and absence of 2.5 μM of ATV. (C) The ATV and GRL0617 (positive control) activity on 100 nM PLpro velocity at 0–10 μM of inhibitor. (D) The 3D representation of the best docking pose for ATV into Mpro catalytic site in the presence of the catalytic water (H2Ocat). For better interpretation the Mpro structure was represented only in the monomeric form with the domains I, II and III in light red, orange, and gray, respectively. Full article ">Figure 3
Pharmacokinetics of atazanavir (ATV) in mice. Swiss–Webster mice at 8–15 weeks of age were orally treated with 60 mg/kg of ATV. (A) At indicated time points, concentration of ATV was measured in the plasma and in the lungs. The insert in panel A represents the pharmacokinetic parameters in the plasma. (B) The area under the curve (AUC) for the anatomical compartments were registered. Full article ">Figure 4
ATV protected K18-hACE2-transgenic mice infected with SARS-CoV-2 from mortality. The survival (A), and weight variation (B) of the different experimental groups: non-infected and non-treated (MOCK) or SARS-CoV-2-infected and non-treated (NIL) SARS-CoV-2-infected and treated with ATV. After 12h of infection the treated group received the first of a daily dose of 60 mg/kg of ATV. The animals were treated for six days. Survival was statistically assessed by Log-rank (Mentel–Cox) test, where * p < 0.05. The viral load (C), LDH levels (D), polymorphonuclear and mononuclear cells counts (E) and immunocytochemical staining (F) were assessed in the BAL six days after infection in the indicated experimental groups. All the analysis were performed with 6 mice/group. Full article ">Figure 5
The proinflammatory content in terms of cytokines IL-6, TNF-α, KC and PF4 in BAL and lung samples. * p < 0.05. Full article ">Figure 6
Microphotographs for histology of lung lobe, bronchiole and alveoli samples from K18-hACE2-transgenic mice non-infected (MOCK) and infected with SARS-CoV-2 gamma strain without (NIL) and treated with ATV upon the six treated days. Full article ">

15 pages, 1281 KiB

Open AccessArticle

Quality Assessment of Investigational Medicinal Products in COVID-19 Clinical Trials: One Year of Activity at the Clinical Trials Office

by Diego Alejandro Dri, Giulia Praticò, Elisa Gaucci, Carlotta Marianecci and Donatella Gramaglia

Pharmaceuticals 2021, 14(12), 1321; https://doi.org/10.3390/ph14121321 - 17 Dec 2021

Cited by 2 | Viewed by 3119

Abstract

One year after the spread of the pandemic, we analyzed the assessment results of the quality documentation submitted to the Clinical Trials Office of the Italian Medicines Agency as part of the request for authorization of clinical trials with a COVID-19 indication. In [...] Read more.

One year after the spread of the pandemic, we analyzed the assessment results of the quality documentation submitted to the Clinical Trials Office of the Italian Medicines Agency as part of the request for authorization of clinical trials with a COVID-19 indication. In this article, we report the classification of the documentation type, an overview of the assessment results, and the related issues focusing on the most frequently detected ones. Relevant data regarding the Investigational Medicinal Products (IMPs) tested in COVID-19 clinical trials and their quality profiles are provided in the perspective of increasing transparency and availability of information. Some criticalities that have been exacerbated by the management of clinical trials during the emergency period are highlighted. Results confirm that IMPs tested in authorized COVID-19 clinical trials are developed in agreement with the same legal requirements for quality, safety, and efficacy as for any other medicinal product in the European Union (EU). The same strong regulatory framework applies, and there is no lowering in the safety profile due to the pandemic; authorized IMPs meet the highest standards of quality. The regulatory network should capitalize on lessons learned from the emergency setting. Some take-home messages are provided that could support the regulatory framework to expand its boundaries by innovating and evolving even though remaining strong and effective. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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28 pages, 4744 KiB

Open AccessArticle

Promising Antiviral Activity of Agrimonia pilosa Phytochemicals against Severe Acute Respiratory Syndrome Coronavirus 2 Supported with In Vivo Mice Study

by Nashwah G. M. Attallah, Aya H. El-Kadem, Walaa A. Negm, Engy Elekhnawy, Thanaa A. El-Masry, Elshaymaa I. Elmongy, Najla Altwaijry, Ashwag S. Alanazi, Gadah Abdulaziz Al-Hamoud and Amany E. Ragab

Pharmaceuticals 2021, 14(12), 1313; https://doi.org/10.3390/ph14121313 - 16 Dec 2021

Cited by 31 | Viewed by 3932

Abstract

The global emergence of the COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has focused the entire world’s attention toward searching for a potential remedy for this disease. Thus, we investigated the antiviral activity of Agrimonia pilosa ethanol extract (APEE) [...] Read more.

The global emergence of the COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has focused the entire world’s attention toward searching for a potential remedy for this disease. Thus, we investigated the antiviral activity of Agrimonia pilosa ethanol extract (APEE) against SARS-CoV-2 and it exhibited a potent antiviral activity with IC₅₀ of 1.1 ± 0.03 µg/mL. Its mechanism of action was elucidated, and it exhibited a virucidal activity and an inhibition of viral adsorption. Moreover, it presented an immunomodulatory activity as it decreased the upregulation of gene expression of COX-2, iNOS, IL-6, TNF-α, and NF-κB in lipopolysaccharide (LPS)-induced peripheral blood mononuclear cells. A comprehensive analysis of the phytochemical fingerprint of APEE was conducted using LC-ESI-MS/MS technique for the first time. We detected 81 compounds and most of them belong to the flavonoid and coumarin classes. Interestingly, isoflavonoids, procyanidins, and anthocyanins were detected for the first time in A. pilosa. Moreover, the antioxidant activity was evidenced in DPPH (IC₅₀ 62.80 µg/mL) and ABTS (201.49 mg Trolox equivalents (TE)/mg) radical scavenging, FRAP (60.84 mg TE/mg), and ORAC (306.54 mg TE/g) assays. Furthermore, the protective effect of APEE was investigated in Lipopolysaccharides (LPS)-induced acute lung injury (ALI) in mice. Lung W/D ratio, serum IL-6, IL-18, IL-1β, HO-1, Caspase-1, caspase-3, TLR-4 expression, TAC, NO, MPO activity, and histopathological examination of lung tissues were assessed. APEE induced a marked downregulation in all inflammation, oxidative stress, apoptosis markers, and TLR-4 expression. In addition, it alleviated all histopathological abnormalities confirming the beneficial effects of APEE in ALI. Therefore, APEE could be a potential source for therapeutic compounds that could be investigated, in future preclinical and clinical trials, in the treatment of patients with COVID-19. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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15 pages, 1252 KiB

Open AccessArticle

In Vitro Effect of Taraxacum officinale Leaf Aqueous Extract on the Interaction between ACE2 Cell Surface Receptor and SARS-CoV-2 Spike Protein D614 and Four Mutants

by Hoai Thi Thu Tran, Michael Gigl, Nguyen Phan Khoi Le, Corinna Dawid and Evelyn Lamy

Pharmaceuticals 2021, 14(10), 1055; https://doi.org/10.3390/ph14101055 - 17 Oct 2021

Cited by 15 | Viewed by 31900

Abstract

To date, there have been rapidly spreading new SARS-CoV-2 “variants of concern”. They all contain multiple mutations in the ACE2 receptor recognition site of the spike protein, compared to the original Wuhan sequence, which is of great concern, because of their potential for [...] Read more.

To date, there have been rapidly spreading new SARS-CoV-2 “variants of concern”. They all contain multiple mutations in the ACE2 receptor recognition site of the spike protein, compared to the original Wuhan sequence, which is of great concern, because of their potential for immune escape. Here we report on the efficacy of common dandelion (Taraxacum officinale) to block protein–protein interaction of SARS-COV-2 spike to the human ACE2 receptor. This could be shown for the wild type and mutant forms (D614G, N501Y, and a mix of K417N, E484K, and N501Y) in human HEK293-hACE2 kidney and A549-hACE2-TMPRSS2 lung cells. High-molecular-weight compounds in the water-based extract account for this effect. Infection of the lung cells using SARS-CoV-2 spike D614 and spike Delta (B.1.617.2) variant pseudotyped lentivirus particles was efficiently prevented by the extract and so was virus-triggered pro-inflammatory interleukin 6 secretion. Modern herbal monographs consider the usage of this medicinal plant as safe. Thus, the in vitro results reported here should encourage further research on the clinical relevance and applicability of the extract as prevention strategy for SARS-CoV-2 infection in terms of a non-invasive, oral post-exposure prophylaxis. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Metabolic analysis of T. officinale (TO) and C. intybus (CI) leaf extract using UPLC-TOF-MS. Measurements were done in high resolution mode with negative electrospray ionization (ESI−) and positive electrospray ionization (ESI+). Full article ">Figure 2
Effect of T. officinale and C. intybus extracts on SARS-CoV-2-Spike–ACE 2 inhibition. (A,B) Concentration-dependent effect of T. officinale (TO) and C. intybus (CI) extract. (C,D). Effect of fractions from TO and CI leaf extract. The extracts were freeze-dried and a molecular weight fractionation was subsequently carried out. The cut-off was set to 5 kDa (HMW > 5 kDa, LMW < 5kDa). H+L: HMW and LMW fractions; 50 mg of dried leaves per mL water was used as reference. HMW and LMW fraction quantities equivalent to dried leaves were used. The binding inhibition was assessed using ELISA technique. Bars are means + SD. Solvent control: distilled water (a.d.); * p < 0.05, ** p < 0.01. Significance of difference was calculated relative to the solvent control by one-way ANOVA. Full article ">Figure 3
Binding inhibition of S1 spike protein to human HEK293-hACE2 cells by extract pre-incubation. Cells were pre-incubated for the indicated times with 10 mg/mL T. officinale (TO), its HMW fraction, equal to 10 mg/mL extract (HMW), and 10 mg/mL C. intybus (CI) or solvent control (a.d.) and subsequently treated with His-tagged S1 spike protein for 1 h without a washing step in between at 4 °C. Binding inhibition was assessed using flow cytometry. N = 3, bars are means + SD. Upper left: cytogram of gated HEK-hACE2 cells. Middle: overlay of representative fluorescence intensity histograms for ACE2 surface expression. Upper right: overlay of representative fluorescence intensity histograms for spike-binding inhibition by the extracts or a.d.; positive control: 20 µg/mL soluble hACE2. Cells were stained with anti-His-tag Alexa Fluor 647 conjugated monoclonal antibody; ** p < 0.01. Significance of difference was calculated relative to the solvent control by one-way ANOVA. Full article ">Figure 4
Binding inhibition of spike D614, and its mutants D614G, N501Y or mix (N501Y, K417N and E484K) to human HEK293-hACE2 and A549-hACE2-TMPRSS2 cells by extract pre- or post-incubation. Overlay of fluorescence intensity histogram for (A) unstained HEK cells, staining control (anti-His-tag Alexa Fluor 647), and cells incubated with His-tag-labelled spike D614, D614G or N501Y for 1 h at 4 °C. (B,C) cells pre-incubated with solvent control (a.d.), 10 mg/mL T. officinale (TO) or 10 mg/mL C. intybus (CI) for 30–60 s, and then treated with His-tag-labelled S1 spike D614, D614G or N501Y protein for 1 h without a washing step in between at 4 °C. (D–G) Effect of extract incubation on HEK or A549 cells either before or after incubation with His-tag-labelled spike D614, D614G, N501Y or mix (N501Y, K417N and E484K) protein at 37 °C. (H) Plant extracts were incubated in saliva from four human donors for 0.5 h at 37 °C. Afterwards, cells were pre-treated with 5 mg/mL extracts for 60 s at 37 °C before incubation with His-tag-labelled spike D614 protein for 0.5 h at 37 °C. Spike-binding inhibition to human cells was assessed using flow cytometric analysis of cells stained with anti-His-tag Alexa Fluor 647 conjugated monoclonal antibody. Bars are means +SD; * p < 0.05, ** p < 0.01. Significance of difference was calculated relative to the respective solvent control by one-way ANOVA. Full article ">Figure 5
Effect of T. officinale extract on ACE2 enzyme activity and protein expression. (A) Viability of A549-hACE2-TMPRSS2 cells was determined using trypan blue cell staining after 84 h exposure to the extract. (B) Cells were incubated with TO extract or 500 ng/mL S1 protein and analyzed for enzyme activity using a fluorescence kit. (C,D) Cells were exposed for 6 h or 24 h to extract without (white bars) or with (black bars) 500 ng/mL S1 protein and analyzed for ACE2 protein expression using a human ACE2 ELISA kit; a.d.: solvent control. Bars are means + SD, N ≥ 3 independent experiments; * p < 0.05, ** p < 0.01. Significance of difference was calculated relative to the respective control by one-way ANOVA. Full article ">Figure 6
Viral transduction inhibition of A549-hACE2-TMPRSS2 cells by T. officinale extract. (A) Cells were pre-treated with T. officinale (TO) or HMW extract for 0.5 h before infection with 7500 TU/mL SARS-CoV-2 spike D614 or Delta (B.1.617.2) variant for 24 h; (B,C) Cells were transduced with 7500 TU/mL SARS-CoV-2 for (B) 3 h before addition of TO for another 21 h or (C) 24 h. After transduction, the medium was exchanged with fresh medium containing TO or HMW extract at the indicated concentrations and post-incubated for 60 h. (D) Cells were pre-treated with 40 mg/mL TO for 3 h before transduction with the indicated virus titer for 24 h. After that, the medium was exchanged with fresh medium and incubated for another 48 h. Luminescence was then detected within 1 h. 0.35 mg/mL HMW extract equals to 20 mg/mL TO extract. Transduction control: (−) negative control: bald lentiviral pseudovirion; (+) positive control: firefly luciferase lentivirus; inhibitor positive control: 100 µg/mL anti-hACE2 antibody. (E) Pro-inflammatory IL-6 cytokine secretion analysis was done either after 24 h virus transduction together with extract (left), after 24 h + 60 h post-infection with extract (middle) or after 60 h post-infection with extract (right) using multiplexing flow cytometric analysis. Solvent control: distilled water (a.d.). N ≥ 3 independent experiments; * p < 0.05, ** p < 0.01. Significance of difference was calculated relative to the solvent control by one-way ANOVA. Full article ">

19 pages, 4584 KiB

Open AccessArticle

SARS-CoV-2 Fears Green: The Chlorophyll Catabolite Pheophorbide A Is a Potent Antiviral

by Guillermo H. Jimenez-Aleman, Victoria Castro, Addis Londaitsbehere, Marta Gutierrez-Rodríguez, Urtzi Garaigorta, Roberto Solano and Pablo Gastaminza

Pharmaceuticals 2021, 14(10), 1048; https://doi.org/10.3390/ph14101048 - 15 Oct 2021

Cited by 16 | Viewed by 6810

Abstract

SARS-CoV-2 pandemic is having devastating consequences worldwide. Although vaccination advances at good pace, effectiveness against emerging variants is unpredictable. The virus has displayed a remarkable resistance to treatments and no drugs have been proved fully effective against COVID-19. Thus, despite the international efforts, [...] Read more.

SARS-CoV-2 pandemic is having devastating consequences worldwide. Although vaccination advances at good pace, effectiveness against emerging variants is unpredictable. The virus has displayed a remarkable resistance to treatments and no drugs have been proved fully effective against COVID-19. Thus, despite the international efforts, there is still an urgent need for new potent and safe antivirals against SARS-CoV-2. Here, we exploited the enormous potential of plant metabolism using the bryophyte Marchantia polymorpha L. and identified a potent SARS-CoV-2 antiviral, following a bioactivity-guided fractionation and mass-spectrometry approach. We found that the chlorophyll derivative Pheophorbide a (PheoA), a porphyrin compound similar to animal Protoporphyrin IX, has an extraordinary antiviral activity against SARS-CoV-2, preventing infection of cultured monkey and human cells, without noticeable cytotoxicity. We also show that PheoA targets the viral particle, interfering with its infectivity in a dose- and time-dependent manner. Besides SARS-CoV-2, PheoA also displayed a broad-spectrum antiviral activity against enveloped RNA viral pathogens such as HCV, West Nile, and other coronaviruses. Our results indicate that PheoA displays a remarkable potency and a satisfactory therapeutic index, which together with its previous use in photoactivable cancer therapy in humans, suggest that it may be considered as a potential candidate for antiviral therapy against SARS-CoV-2. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Graphical abstract
Full article ">Figure 1
M. polymorpha extracts interfere with SARS-CoV-2-induced cytopathic effect and virus propagation. (A,B) Vero E6 cells were inoculated with SARS-CoV-2 (MOI = 0.001) in the presence of serial 2-fold dilutions of crude extracts from (A) two different M. polymorpha ecotypes, ruderalis (Ex1) and polymorpha (Ex2) or (B) WT, Mpcoi1-2 or Mpc1hdz Marchantia plants, incubated for 72 h, time after which they were fixed and stained with a crystal violet solution. Mock-infected cells were used as the control of the integrity of the cell monolayer. Images show a representative experimental plate comparing M. polymorpha extracts with vehicle (DMSO)-treated cells. (C) Vero E6 cells were inoculated with SARS-CoV-2 (MOI = 0.001) in the presence of vehicle (DMSO), RMDV (25 µM) or a 100 µg/mL dilution of crude extracts. Uninfected samples were used as the control (mock). Total RNA was extracted 72 h post-infection and subjected to RT-qPCR to determine viral load. The dotted line indicates the limit of detection of the assay. Normalized viral RNA levels are shown as percentage of the viral RNA found in vehicle-treated cells. Data are shown as mean (± SD) of three biological replicates. Statistical significance was estimated using one-way ANOVA and a Dunnet’s post hoc test (* p < 0.05). Full article ">Figure 2
Extract fractionation and identification of antiviral fraction pools. (A) Vero E6 cells were inoculated with SARS-CoV-2 (MOI = 0.001) in the presence of serial 2-fold dilutions of vehicle (DMSO), a crude Marchantia extract and the fraction pools. Inoculated cultures were incubated for 72 h, time after which they were fixed and stained with a crystal violet solution. Mock-treated cells were used as the control of the integrity of the cell monolayer (non-infected). (B) Vero E6 cells were inoculated (MOI = 0.01) in the presence of serial dilution of the fractions and incubated for 24 h before fixation; processing for immunofluorescence microscopy and cytotoxicity assays was as described in the materials and methods section. Representative images of the fraction pool cell-based evaluation are shown. (C) Quantitation of infection efficiency, cell number and cell viability. These data are shown as average (± SD) of three biological replicates. Statistical significance was estimated using one-way ANOVA (Dunnet´s post hoc test, * p < 0.05). Full article ">Figure 3
Identification of the main antiviral compound in Marchantia extracts. (A) Representative TLC of Marchantia WT and c1hdz extracts. Compounds C, D and E (box) were tested for their antiviral potential. (B) Vero E6 cells were inoculated (MOI = 0.01) in the presence of the indicated compounds and incubated for 24 h before fixation and processing for immunofluorescence microscopy. Data are shown as average (± SD) of three biological replicates. Statistical significance was estimated using one-way ANOVA and a Dunnet´s post hoc test (* p < 0.05). (C) Representative HPLC/MS analysis (shown for WT) of fractions C, D, and E, including exact mass determination of the antiviral candidate 1, and its inferred chemical structure. Full article ">Figure 4
PheoA shows antiviral activity against SARS-CoV-2 in Vero E6 cells and human lung epithelial A549-ACE2 and Calu3 cell lines. (A–C) Commercially available PheoA was serially diluted and mixed (1:1, v/v) with SARS-CoV-2 preparations to achieve the indicated compound concentrations and a final MOI of 0.005 for (A) Vero E6 and (B) Calu3 and 0.01 for (C) A549-ACE2 cells. Cultures were incubated for 48 h, fixed and processed for automated immunofluorescence microscopy analysis. Parallel, uninfected cultures were processed for cytotoxicity evaluation using an MTT assay. Relative infection efficiency data (n = 3 per dose) are shown as individual data and a PROBIT regression curve (green line) using the represented values. Cytotoxicity data (n = 3 per dose) are shown as the individual data and a moving average trend line. (D) A549-ACE2 cells were inoculated at MOI = 0.01 in the presence of increasing concentrations of PheoA or RMDV (5 µM) and incubated for 48 h. Samples of the supernatants were collected, heat-inactivated and directly subjected to RT-qPCR to estimate overall infection efficiency. Data are expressed as relative values compared with the vehicle (DMSO)-treated cells and are shown as the mean (± SD) of three biological replicates (n = 3). Statistical significance was estimated using one-way ANOVA and a Dunnet´s post hoc test (* p < 0.05). Full article ">Figure 5
Antiviral spectrum of PheoA against different RNA viruses. The effectiveness of PheoA was tested against four different recombinant RNA viruses expressing reporter genes. (A) Huh7 cells were infected with HCVtcp in the presence of increasing PheoA doses and luciferase activity was determined 48 h post-inoculation. (B–D) Cells were inoculated in the presence of increasing concentrations of PheoA at MOI 0.01 and incubated to enable virus propagation. At the endpoint, cells were fixed and counter stained with DAPI to control for unexpected cytotoxic effects. Relative infection efficiency was estimated using automated microscopy and is expressed as percentage of the infection efficiency observed in control wells. (B) Huh7 cells were infected with hCoV-229E-GFP and fixed 48 h post-inoculation. (C) Huh7 cells were infected with WNV-GFP and fixed 48 h post-inoculation. (D) A549-ACE2 cells were inoculated with VSV-GFP and fixed 16 h post-inoculation. Individual replicate data are shown as green dots (n = 3) and the PROBIT regression curve used to estimate EC50 values is shown. Full article ">Figure 6
Combination treatment of PheoA with RMDV. Vero E6 cells were inoculated at MOI = 0.005 in the presence of increasing concentrations of PheoA in combination with increasing doses of RMDV. Twenty-four hours post infection, cells were fixed and processed for automated immunofluorescence microscopy. Relative infection efficiency values were estimated as percentage of the values obtained in mock-treated cells. (A) Data are shown as average of two biological replicates. (B) Heatmap describing the areas of synergy within the combination treatments. Full article ">Figure 7
Time-of-addition experiments indicate that PheoA interferes with early aspects of SARS-CoV-2 infection. Vero E6 cells were inoculated at MOI = 5 in the presence (gray) or absence (white) of the indicated doses of PheoA, RMDV or imatinib as described in both the text and the scheme. Cells were incubated for 6 h in the presence (gray) or absence (white) before chemical fixation and processing for immunofluorescence microscopy. (A) Schematic diagram of the times where compound was present in the assay. (B) Infection efficiency is expressed as the percentage of that observed in vehicle DMSO-treated cells and is shown as average and standard deviation of three biological replicates (n = 3). Statistical significance was estimated using one-way ANOVA and a Dunnet´s post hoc test (* p < 0.05). Full article ">Figure 8
PheoA interferes with SARS-CoV-2 virion infectivity in a dose- and time-dependent manner. SARS-CoV-2 virus stocks were diluted to obtain 1 × 105 TCID50/mL and were mixed with increasing concentrations of PheoA or the vehicle (DMSO). (A) Dose-dependent reduction in SARS-CoV-2 infectivity by PheoA. Virus-compound mixtures were incubated at room temperature for 30 min and were serially diluted to determine the remaining infectivity titer using endpoint dilution and determination of virus-induced cytopathic effect by crystal violet staining in Vero-E6 cells. (B) Pre-incubation time-dependent reduction in SARS-CoV-2 infectivity by PheoA. Experiments were carried out using a fixed dose of PheoA (340 nM) and increasing pre-incubation times before serial dilution for TCID50 determination. Values are expressed as LOG TCID50/mL and shown as the average and standard deviation of three independent experiments (n = 3). Statistical significance was estimated using one-way ANOVA and a Dunnet’s post hoc test (* p < 0.05). Full article ">

13 pages, 3311 KiB

Open AccessArticle

A Drug Repurposing Approach for Antimalarials Interfering with SARS-CoV-2 Spike Protein Receptor Binding Domain (RBD) and Human Angiotensin-Converting Enzyme 2 (ACE2)

by Paolo Coghi, Li Jun Yang, Jerome P. L. Ng, Richard K. Haynes, Maurizio Memo, Alessandra Gianoncelli, Vincent Kam Wai Wong and Giovanni Ribaudo

Pharmaceuticals 2021, 14(10), 954; https://doi.org/10.3390/ph14100954 - 23 Sep 2021

Cited by 18 | Viewed by 4957

Abstract

Host cell invasion by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is mediated by the interaction of the viral spike protein (S) with human angiotensin-converting enzyme 2 (ACE2) through the receptor-binding domain (RBD). In this work, computational and experimental techniques were combined to [...] Read more.

Host cell invasion by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is mediated by the interaction of the viral spike protein (S) with human angiotensin-converting enzyme 2 (ACE2) through the receptor-binding domain (RBD). In this work, computational and experimental techniques were combined to screen antimalarial compounds from different chemical classes, with the aim of identifying small molecules interfering with the RBD-ACE2 interaction and, consequently, with cell invasion. Docking studies showed that the compounds interfere with the same region of the RBD, but different interaction patterns were noted for ACE2. Virtual screening indicated pyronaridine as the most promising RBD and ACE2 ligand, and molecular dynamics simulations confirmed the stability of the predicted complex with the RBD. Bio-layer interferometry showed that artemisone and methylene blue have a strong binding affinity for RBD (K_D = 0.363 and 0.226 μM). Pyronaridine also binds RBD and ACE2 in vitro (K_D = 56.8 and 51.3 μM). Overall, these three compounds inhibit the binding of RBD to ACE2 in the μM range, supporting the in silico data. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Full article ">Figure 1
Structures of the antimalarial compounds used in this investigation. Full article ">Figure 2
Calculated binding energies for the interaction of the compounds with RBD (PDB ID: 6VSB) and ACE2 (PDB ID: 6LZG). Full article ">Figure 3
(a) Predicted binding motif for the compounds with RBD; (b) detailed view of the residues interacting with pyronaridine. Full article ">Figure 4
Predicted binding motif for the compounds with ACE2 (a). Detailed view of the residues interacting with pyronaridine (b). Full article ">Figure 5
MD simulations for the complexes of pyronaridine with (a) RBD and (b) ACE2 obtained from docking experiments. Root mean square deviation (RMSD) is depicted in green for the ligand and in dark grey for the protein. Full article ">Figure 6
Binding kinetics and affinity analysis for the interaction of pyronaridine with RBD (a) and ACE2 (b) according to bio-layer interferometry studies. Full article ">Figure 7
Results of ELISA experiments demonstrating that pyronaridine, artemisone and methylene blue inhibit the binding of RBD to ACE2 (* p < 0.05, compared with control group). Full article ">

14 pages, 3419 KiB

Open AccessArticle

Scaffold Hopping of α-Rubromycin Enables Direct Access to FDA-Approved Cromoglicic Acid as a SARS-CoV-2 M^Pro Inhibitor

by Hani A. Alhadrami, Ahmed M. Sayed, Heba Al-Khatabi, Nabil A. Alhakamy and Mostafa E. Rateb

Pharmaceuticals 2021, 14(6), 541; https://doi.org/10.3390/ph14060541 - 5 Jun 2021

Cited by 20 | Viewed by 4791

Abstract

The COVID-19 pandemic is still active around the globe despite the newly introduced vaccines. Hence, finding effective medications or repurposing available ones could offer great help during this serious situation. During our anti-COVID-19 investigation of microbial natural products (MNPs), we came across α-rubromycin, [...] Read more.

The COVID-19 pandemic is still active around the globe despite the newly introduced vaccines. Hence, finding effective medications or repurposing available ones could offer great help during this serious situation. During our anti-COVID-19 investigation of microbial natural products (MNPs), we came across α-rubromycin, an antibiotic derived from Streptomyces collinus ATCC19743, which was able to suppress the catalytic activity (IC₅₀ = 5.4 µM and K_i = 3.22 µM) of one of the viral key enzymes (i.e., M^Pro). However, it showed high cytotoxicity toward normal human fibroblasts (CC₅₀ = 16.7 µM). To reduce the cytotoxicity of this microbial metabolite, we utilized a number of in silico tools (ensemble docking, molecular dynamics simulation, binding free energy calculation) to propose a novel scaffold having the main pharmacophoric features to inhibit M^Pro with better drug-like properties and reduced/minimal toxicity. Nevertheless, reaching this novel scaffold synthetically is a time-consuming process, particularly at this critical time. Instead, this scaffold was used as a template to explore similar molecules among the FDA-approved medications that share its main pharmacophoric features with the aid of pharmacophore-based virtual screening software. As a result, cromoglicic acid (aka cromolyn) was found to be the best hit, which, upon in vitro M^Pro testing, was 4.5 times more potent (IC₅₀ = 1.1 µM and K_i = 0.68 µM) than α-rubromycin, with minimal cytotoxicity toward normal human fibroblasts (CC₅₀ > 100 µM). This report highlights the potential of MNPs in providing unprecedented scaffolds with a wide range of therapeutic efficacy. It also revealed the importance of cheminformatics tools in speeding up the drug discovery process, which is extremely important in such a critical situation. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Figure 1
(A) Monomeric structure of SARS CoV-2 MPro (PDB: 6LU7) showing its three main domains (I, II, and III; blue, green, and orange, respectively). (B) MPro active site showing the catalytic dyad (HIS41–CYS145). (C) The dimeric active form of SARS CoV-2 MPro. Full article ">Figure 2
The general outline of the present investigation illustrating the main steps starting from α-rubromycin and eventually reaching cromoglicic acid. Full article ">Figure 3
Binding orientations of α-rubromycin inside the MPro active site (orientations 1–3, A–C respectively), and their average RMSDs over 50 ns of MDS (D). Full article ">Figure 4
Binding modes of (A) α-rubromycin, (B) ScafA, and (C) cromoglicic acid inside the MPro active site. (D) RMSDs of their docking pose over 200 ns of MDS. Full article ">Figure 5
Protein–ligand contacts inside the MPro active sites over 200 ns of MDS: (A) α-rubromycin, (B) ScafA, and (C) cromoglicic acid. Full article ">Figure 6
Docking pose alignment of α-rubromycin (green color), ScafA (red color), and cromoglicic acid (cyan color). This alignment shows that the three compounds have a good alignment and share the same pharmacophoric features. Full article ">

20 pages, 8798 KiB

Open AccessArticle

Identification of Anti-SARS-CoV-2 Compounds from Food Using QSAR-Based Virtual Screening, Molecular Docking, and Molecular Dynamics Simulation Analysis

by Magdi E. A. Zaki, Sami A. Al-Hussain, Vijay H. Masand, Siddhartha Akasapu, Sumit O. Bajaj, Nahed N. E. El-Sayed, Arabinda Ghosh and Israa Lewaa

Pharmaceuticals 2021, 14(4), 357; https://doi.org/10.3390/ph14040357 - 13 Apr 2021

Cited by 27 | Viewed by 5000

Abstract

Due to the genetic similarity between SARS-CoV-2 and SARS-CoV, the present work endeavored to derive a balanced Quantitative Structure−Activity Relationship (QSAR) model, molecular docking, and molecular dynamics (MD) simulation studies to identify novel molecules having inhibitory potential against the main protease (Mpro) of [...] Read more.

Due to the genetic similarity between SARS-CoV-2 and SARS-CoV, the present work endeavored to derive a balanced Quantitative Structure−Activity Relationship (QSAR) model, molecular docking, and molecular dynamics (MD) simulation studies to identify novel molecules having inhibitory potential against the main protease (Mpro) of SARS-CoV-2. The QSAR analysis developed on multivariate GA–MLR (Genetic Algorithm–Multilinear Regression) model with acceptable statistical performance (R² = 0.898, Q²loo = 0.859, etc.). QSAR analysis attributed the good correlation with different types of atoms like non-ring Carbons and Nitrogens, amide Nitrogen, sp²-hybridized Carbons, etc. Thus, the QSAR model has a good balance of qualitative and quantitative requirements (balanced QSAR model) and satisfies the Organisation for Economic Co-operation and Development (OECD) guidelines. After that, a QSAR-based virtual screening of 26,467 food compounds and 360 heterocyclic variants of molecule 1 (benzotriazole–indole hybrid molecule) helped to identify promising hits. Furthermore, the molecular docking and molecular dynamics (MD) simulations of Mpro with molecule 1 recognized the structural motifs with significant stability. Molecular docking and QSAR provided consensus and complementary results. The validated analyses are capable of optimizing a drug/lead candidate for better inhibitory activity against the main protease of SARS-CoV-2. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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18 pages, 2934 KiB

Open AccessArticle

Host-Directed FDA-Approved Drugs with Antiviral Activity against SARS-CoV-2 Identified by Hierarchical In Silico/In Vitro Screening Methods

by Tiziana Ginex, Urtzi Garaigorta, David Ramírez, Victoria Castro, Vanesa Nozal, Inés Maestro, Javier García-Cárceles, Nuria E. Campillo, Ana Martinez, Pablo Gastaminza and Carmen Gil

Pharmaceuticals 2021, 14(4), 332; https://doi.org/10.3390/ph14040332 - 6 Apr 2021

Cited by 23 | Viewed by 6161

Abstract

The unprecedent situation generated by the COVID-19 global emergency has prompted us to actively work to fight against this pandemic by searching for repurposable agents among FDA approved drugs to shed light into immediate opportunities for the treatment of COVID-19 patients. In the [...] Read more.

The unprecedent situation generated by the COVID-19 global emergency has prompted us to actively work to fight against this pandemic by searching for repurposable agents among FDA approved drugs to shed light into immediate opportunities for the treatment of COVID-19 patients. In the attempt to proceed toward a proper rationalization of the search for new antivirals among approved drugs, we carried out a hierarchical in silico/in vitro protocol which successfully combines virtual and biological screening to speed up the identification of host-directed therapies against COVID-19 in an effective way. To this end a multi-target virtual screening approach focused on host-based targets related to viral entry, followed by the experimental evaluation of the antiviral activity of selected compounds, has been carried out. As a result, five different potentially repurposable drugs interfering with viral entry—cepharantine, clofazimine, metergoline, imatinib and efloxate—have been identified. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Figure 1
Schematic representation of the eight targets selected in this study and their role in virus entry. Representative inhibitors are also cited, when available. Full article ">Figure 2
Schematic representation of the computational protocol applied in this study. For each target, green and blue circles respectively mark the active and the allosteric/secondary binding sites. Full article ">Figure 3
Potency and cytotoxicity indexes of the confirmed primary hits. Potency (EC50; EC90) and cytotoxicity (CC50) were calculated from dose-response experiments in which infection efficiency was determined by viral antigen accumulation and MTT activity respectively (see <a href="#app1-pharmaceuticals-14-00332" class="html-app">Figures S2 and S3 of the Supplementary Materials</a>). Remdesivir was included as control of the assays. Full article ">Figure 4
Antiviral candidates interfere with viral entry of SARS-CoV-2 pseudotypes. (A) Retroviral vectors pseudotyped with SARS-CoV-2 Spike glycoprotein were used to inoculate Vero-E6 cells in the presence of the candidates at the EC90 (see <a href="#pharmaceuticals-14-00332-f003" class="html-fig">Figure 3</a>). Forty-eight hours later, total cell lysates were assayed to determine luciferase activity as a reporter activity for viral entry. (B) Compound selectivity was assayed also using VSVpp and RD114pp for the compounds interfering with Spp. Data are shown as average and SEM of a minimum of four biological replicates (N = 4). Statistical significance was evaluated using a one-way ANOVA and a Dunnet’s post-hoc test (* p < 0.05). Full article ">Figure 5
Heat matrix showing the potential targets for the 13 confirmed positive FDA compounds from <a href="#app1-pharmaceuticals-14-00332" class="html-app">Table S4</a> based on their MM/GBSA scores. The five finally identified as entry inhibitors are highlighted in red. Full article ">Figure 6
Binding modes and relative MMGBSA scores for (A) cepharantine (CET) in complex with TPC2, (B) efloxate (EFX) in complex with AAK1, (C) metergoline (MTG) in complex with cathepsin L and (D) clofazimine (CFZ) in complex with GAK. Full article ">

11 pages, 1131 KiB

Open AccessArticle

Association between Functional Inhibitors of Acid Sphingomyelinase (FIASMAs) and Reduced Risk of Death in COVID-19 Patients: A Retrospective Cohort Study

by Gil Darquennes, Pascal Le Corre, Olivier Le Moine and Gwenolé Loas

Pharmaceuticals 2021, 14(3), 226; https://doi.org/10.3390/ph14030226 - 7 Mar 2021

Cited by 24 | Viewed by 4319

Abstract

Given the current scarcity of curative treatment of COVID-19, the search for an effective treatment modality among all available medications has become a priority. This study aimed at investigating the role of functional inhibitors of acid sphingomyelinase (FIASMAs) on in-hospital COVID-19 mortality. In [...] Read more.

Given the current scarcity of curative treatment of COVID-19, the search for an effective treatment modality among all available medications has become a priority. This study aimed at investigating the role of functional inhibitors of acid sphingomyelinase (FIASMAs) on in-hospital COVID-19 mortality. In this retrospective cohort study, we included adult in-patients with laboratory-confirmed COVID-19 between 1 March 2020 and 31 August 2020 with definite outcomes (discharged hospital or deceased) from Erasme Hospital (Brussels, Belgium). We used univariate and multivariate logistic regression models to explore the risk factors associated with in-hospital mortality. We included 350 patients (205 males, 145 females) with a mean age of 63.24 years (SD = 17.4, range: 21–96 years). Seventy-two patients died in the hospital and 278 were discharged. The four most common comorbidities were hypertension (184, 52.6%), chronic cardiac disease (110, 31.4%), obesity (96, 27.8%) and diabetes (95, 27.1%). Ninety-three participants (26.6%) received a long-term prescription for FIASMAs. Among these, 60 (64.5%) received amlodipine. For FIASMAs status, multivariable regression showed increasing odds ratio (OR) for in-hospital deaths associated with older age (OR 1.05, 95% CI: 1.02–1.07; p = 0.00015), and higher prevalence of malignant neoplasm (OR 2.09, 95% CI: 1.03–4.22; p = 0.039). Nonsignificant decreasing OR (0.53, 95% CI: 0.27–1.04; p = 0.064) was reported for FIASMA status. For amlodipine status, multivariable regression revealed increasing OR of in-hospital deaths associated with older age (OR 1.04, 95% CI: 1.02–1.07; p = 0.0009), higher prevalence of hypertension (OR 2.78, 95% CI: 1.33–5.79; p = 0.0062) and higher prevalence of malignant neoplasm (OR 2.71, 95% CI: 1.23–5.97; p = 0.013), then secondarily decreasing OR of in-hospital death associated with long-term treatment with amlodipine (OR 0.24, 95% CI: 0.09–0.62; p = 0.0031). Chronic treatment with amlodipine could be significantly associated with low mortality of COVID-19 in-patients. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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15 pages, 3533 KiB

Open AccessArticle

Evaluation of the Antiviral Activity of Sitagliptin-Glatiramer Acetate Nano-Conjugates against SARS-CoV-2 Virus

by Nabil A. Alhakamy, Osama A. A. Ahmed, Tarek S. Ibrahim, Hibah M. Aldawsari, Khalid Eljaaly, Usama A. Fahmy, Ahmed L. Alaofi, Filippo Caraci and Giuseppe Caruso

Pharmaceuticals 2021, 14(3), 178; https://doi.org/10.3390/ph14030178 - 24 Feb 2021

Cited by 14 | Viewed by 3744

Abstract

The outbreak of the COVID-19 pandemic in China has become an urgent health and economic challenge. There is a current race for developing strategies to treat and/or prevent COVID-19 worldwide. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the strain of coronavirus that [...] Read more.

The outbreak of the COVID-19 pandemic in China has become an urgent health and economic challenge. There is a current race for developing strategies to treat and/or prevent COVID-19 worldwide. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the strain of coronavirus that causes COVID-19. The aim of the present work was to evaluate the efficacy of the combined complex (nano-conjugates) of two FDA-approved drugs, sitagliptin (SIT) and glatiramer acetate (GA), against a human isolate of the SARS-CoV-2 virus. SIT-GA nano-conjugates were prepared according to a full three-factor bilevel (2³) factorial design. The SIT concentration (mM, X₁), GA concentration (mM, X₂), and pH (X₃) were selected as the factors. The particle size (nm, Y₁) and zeta potential (mV, Y₂) were assessed as responses. Characterization of the optimized formula for the Fourier-transform infrared (FTIR) spectroscopy and transmission electron microscopy (TEM) was carried out. In addition, the half-maximal inhibitory concentration (IC₅₀) in Vero-E6 epithelial cells previously infected with the virus was investigated. The results revealed that the optimized formula of the prepared complex was a 1:1 SIT:GA molar ratio at a pH of 10, which met the required criteria with a desirability value of 0.878 and had a particle size and zeta potential at values of 77.42 nm and 27.67 V, respectively. The SIT-GA nano-complex showed antiviral potential against an isolate of SARS-CoV-2 with IC50 values of 16.14, 14.09, and 8.52 µM for SIT, GA, and SIT-GA nano-conjugates, respectively. Molecular docking has shown that the formula’s components have a high binding affinity to the COVID 3CL protease, essential for coronavirus replication, paralleled by 3CL protease inhibition (IC₅₀ = 2.87 µM). An optimized formulation of SIT-GA could guarantee both enhanced deliveries to target cells and improved cellular uptake. Further clinical studies are being carried out to validate the clinical efficacy of the optimized formulation against SARS-CoV-2. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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17 pages, 1443 KiB

Open AccessArticle

Identification of 37 Heterogeneous Drug Candidates for Treatment of COVID-19 via a Rational Transcriptomics-Based Drug Repurposing Approach

by Andrea Gelemanović, Tinka Vidović, Višnja Stepanić and Katarina Trajković

Pharmaceuticals 2021, 14(2), 87; https://doi.org/10.3390/ph14020087 - 25 Jan 2021

Cited by 5 | Viewed by 4546

Abstract

A year after the initial outbreak, the COVID-19 pandemic caused by SARS-CoV-2 virus remains a serious threat to global health, while current treatment options are insufficient to bring major improvements. The aim of this study is to identify repurposable drug candidates with a [...] Read more.

A year after the initial outbreak, the COVID-19 pandemic caused by SARS-CoV-2 virus remains a serious threat to global health, while current treatment options are insufficient to bring major improvements. The aim of this study is to identify repurposable drug candidates with a potential to reverse transcriptomic alterations in the host cells infected by SARS-CoV-2. We have developed a rational computational pipeline to filter publicly available transcriptomic datasets of SARS-CoV-2-infected biosamples based on their responsiveness to the virus, to generate a list of relevant differentially expressed genes, and to identify drug candidates for repurposing using LINCS connectivity map. Pathway enrichment analysis was performed to place the results into biological context. We identified 37 structurally heterogeneous drug candidates and revealed several biological processes as druggable pathways. These pathways include metabolic and biosynthetic processes, cellular developmental processes, immune response and signaling pathways, with steroid metabolic process being targeted by half of the drug candidates. The pipeline developed in this study integrates biological knowledge with rational study design and can be adapted for future more comprehensive studies. Our findings support further investigations of some drugs currently in clinical trials, such as itraconazole and imatinib, and suggest 31 previously unexplored drugs as treatment options for COVID-19. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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24 pages, 3883 KiB

Open AccessArticle

FDA-Approved Drugs with Potent In Vitro Antiviral Activity against Severe Acute Respiratory Syndrome Coronavirus 2

by Ahmed Mostafa, Ahmed Kandeil, Yaseen A. M. M. Elshaier, Omnia Kutkat, Yassmin Moatasim, Adel A. Rashad, Mahmoud Shehata, Mokhtar R. Gomaa, Noura Mahrous, Sara H. Mahmoud, Mohamed GabAllah, Hisham Abbas, Ahmed El Taweel, Ahmed E. Kayed, Mina Nabil Kamel, Mohamed El Sayes, Dina B. Mahmoud, Rabeh El-Shesheny, Ghazi Kayali and Mohamed A. Ali

Pharmaceuticals 2020, 13(12), 443; https://doi.org/10.3390/ph13120443 - 4 Dec 2020

Cited by 122 | Viewed by 12432

Abstract

(1) Background: Drug repositioning is an unconventional drug discovery approach to explore new therapeutic benefits of existing drugs. Currently, it emerges as a rapid avenue to alleviate the COVID-19 pandemic disease. (2) Methods: Herein, we tested the antiviral activity of anti-microbial and anti-inflammatory [...] Read more.

(1) Background: Drug repositioning is an unconventional drug discovery approach to explore new therapeutic benefits of existing drugs. Currently, it emerges as a rapid avenue to alleviate the COVID-19 pandemic disease. (2) Methods: Herein, we tested the antiviral activity of anti-microbial and anti-inflammatory Food and Drug Administration (FDA)-approved drugs, commonly prescribed to relieve respiratory symptoms, against Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the viral causative agent of the COVID-19 pandemic. (3) Results: Of these FDA-approved antimicrobial drugs, Azithromycin, Niclosamide, and Nitazoxanide showed a promising ability to hinder the replication of a SARS-CoV-2 isolate, with IC₅₀ of 0.32, 0.16, and 1.29 µM, respectively. We provided evidence that several antihistamine and anti-inflammatory drugs could partially reduce SARS-CoV-2 replication in vitro. Furthermore, this study showed that Azithromycin can selectively impair SARS-CoV-2 replication, but not the Middle East Respiratory Syndrome Coronavirus (MERS-CoV). A virtual screening study illustrated that Azithromycin, Niclosamide, and Nitazoxanide bind to the main protease of SARS-CoV-2 (Protein data bank (PDB) ID: 6lu7) in binding mode similar to the reported co-crystalized ligand. Also, Niclosamide displayed hydrogen bond (HB) interaction with the key peptide moiety GLN: 493A of the spike glycoprotein active site. (4) Conclusions: The results suggest that Piroxicam should be prescribed in combination with Azithromycin for COVID-19 patients. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Figure 1
Dose-inhibition curves for anti-microbial and anti-inflammatory FDA-approved drugs with high selectivity indices against NRC-03-nhCoV. (a) Amikacin sulphate, Azithromycin, Ceftazidime, Doxycycline, Levofloxacin, Moxifloxacin, Niclosamide, and Nitazoxanide, (b) Aspirin, Chlorpheniramine maleate, and Piroxicam. Inhibitory concentration 50% (IC50) values were calculated using nonlinear regression analysis of GraphPad Prism software (version 5.01) by plotting log inhibitor versus normalized response (variable slope). Full article ">Figure 2
Chemical structure of ligands α-ketomaide and N3 for SARS-CoV-2 Mpro (PDB ID: 6lu7, 6y2f), and Ligand 1 for SARS-CoV-2 spike glycoprotein (PDB ID: 6vsb). Full article ">Figure 3
Visual representation by volumetric image display and analysis (VIDA) of docking with Mpro (PDB ID: 6lu7). (a) Azithromycin docked with the formation of two HBs (green color). (b) Niclosamide (grey color) and Nitazoxanide (thiazole ring with yellow-blue color) occupied the active site without detection of HB. Full article ">Figure 4
Visual representation by VIDA for docking with spike glycoprotein (PDB ID: 6vsb). (a) Standard ligand docked inside the receptor (HB in green color), (b) ligand inside the inner grid for validation, (c) Azithromycin docked peripherally, (d) Nitazoxanide docked with formation of weak HB (green color), and (e) Niclosamide docked with formation of strong HB (green color). Full article ">Figure 4 Cont.
Visual representation by VIDA for docking with spike glycoprotein (PDB ID: 6vsb). (a) Standard ligand docked inside the receptor (HB in green color), (b) ligand inside the inner grid for validation, (c) Azithromycin docked peripherally, (d) Nitazoxanide docked with formation of weak HB (green color), and (e) Niclosamide docked with formation of strong HB (green color). Full article ">Figure 5
Differential anti-SARS-CoV-2 and Anti-MERS-CoV activities for Azithromycin, Niclosamide, and Nitazoxanide. (a) Anti-SARS-CoV-2 activity for Azithromycin, Niclosamide, and Nitazoxanide, as measured by Plaque reduction assay. (b) Anti-MERS-CoV activity for Azithromycin, Niclosamide, and Nitazoxanide, as measured by Plaque reduction assay. Full article ">Figure 6
Visual representation by VIDA for docking with the main protease of MERS-CoV (PDB ID: 4ylu). (a) Niclosamide (green color) and Nitazoxanide (grey color) overlay each other, (b) ligand (grey color), Niclosamide (green color), and Nitazoxanide (grey color with yellow sulfur atom color) inside the inner grid for validation, and (c) Azithromycin with amino outside the grid. Full article ">Figure 7
Visual representation by VIDA for docking with the main protease of MERS-CoV (PDB ID: 5x4r). (a) Azithromycin forms HB with ASN:125A, and (b) Niclosamide (grey color) and Nitazoxanide (green color) overlay each other. Full article ">

13 pages, 1279 KiB

Open AccessArticle

Repurposing of Plasminogen: An Orphan Medicinal Product Suitable for SARS-CoV-2 Inhalable Therapeutics

by Anna Maria Piras, Ylenia Zambito, Maurizio Lugli, Baldassare Ferro, Paolo Roncucci, Filippo Mori, Alfonso Salvatore, Ester Ascione, Marta Bellini and Roberto Crea

Pharmaceuticals 2020, 13(12), 425; https://doi.org/10.3390/ph13120425 - 27 Nov 2020

Cited by 4 | Viewed by 3227

Abstract

The SARS-CoV-2 infection is associated with pulmonary coagulopathy, which determines the deposition of fibrin in the air spaces and lung parenchyma. The resulting lung lesions compromise patient pulmonary function and increase mortality, or end in permanent lung damage for those who have recovered [...] Read more.

The SARS-CoV-2 infection is associated with pulmonary coagulopathy, which determines the deposition of fibrin in the air spaces and lung parenchyma. The resulting lung lesions compromise patient pulmonary function and increase mortality, or end in permanent lung damage for those who have recovered from the COVID-19 disease. Therefore, local pulmonary fibrinolysis can be efficacious in degrading pre-existing fibrin clots and reducing the conversion of lung lesions into lasting scars. Plasminogen is considered a key player in fibrinolysis processes, and in view of a bench-to-bedside translation, we focused on the aerosolization of an orphan medicinal product (OMP) for ligneous conjunctivitis: human plasminogen (PLG-OMP) eye drops. As such, the sterile and preservative-free solution guarantees the pharmaceutical quality of GMP production and meets the Ph. Eur. requirements of liquid preparations for nebulization. PLG-OMP aerosolization was evaluated both from technological and stability viewpoints, after being submitted to either jet or ultrasonic nebulization. Jet nebulization resulted in a more efficient delivery of an aerosol suitable for pulmonary deposition. The biochemical investigation highlighted substantial protein integrity maintenance with the percentage of native plasminogen band > 90%, in accordance with the quality specifications of PLG-OMP. In a coherent way, the specific activity of plasminogen is maintained within the range 4.8–5.6 IU/mg (PLG-OMP pre-nebulization: 5.0 IU/mg). This is the first study that focuses on the technological and biochemical aspects of aerosolized plasminogen, which could affect both treatment efficacy and clinical dosage delivery. Increasing evidence for the need of local fibrinolytic therapy could merge with the availability of PLG-OMP as an easy handling solution, readily aerosolizable for a fast translation into an extended clinical efficacy assessment in COVID-19 patients. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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11 pages, 1861 KiB

Open AccessFeature PaperArticle

Effect of Tocilizumab in Hospitalized Patients with Severe COVID-19 Pneumonia: A Case-Control Cohort Study

by Benjamin Rossi, Lee S. Nguyen, Philippe Zimmermann, Faiza Boucenna, Louis Dubret, Louise Baucher, Helene Guillot, Marie-Anne Bouldouyre, Yves Allenbach, Joe-Elie Salem, Paul Barsoum, Arezki Oufella and Helene Gros

Pharmaceuticals 2020, 13(10), 317; https://doi.org/10.3390/ph13100317 - 17 Oct 2020

Cited by 32 | Viewed by 4780

Abstract

Tocilizumab, an anti-interleukin-6 receptor, administrated during the right timeframe may be beneficial against coronavirus-disease-2019 (COVID-19) pneumonia. All patients admitted for severe COVID-19 pneumonia (SpO₂ ≤ 96% despite O₂-support ≥ 6 L/min) without invasive mechanical ventilation were included in a retrospective [...] Read more.

Tocilizumab, an anti-interleukin-6 receptor, administrated during the right timeframe may be beneficial against coronavirus-disease-2019 (COVID-19) pneumonia. All patients admitted for severe COVID-19 pneumonia (SpO₂ ≤ 96% despite O₂-support ≥ 6 L/min) without invasive mechanical ventilation were included in a retrospective cohort study in a primary care hospital. The treatment effect of a single-dose, 400 mg, of tocilizumab was assessed by comparing those who received tocilizumab to those who did not. Selection bias was mitigated using three statistical methods. Primary outcome measure was a composite of mortality and ventilation at day 28. A total of 246 patients were included (106 were treated with tocilizumab). Overall, 105 (42.7%) patients presented the primary outcome, with 71 (28.9%) deaths during the 28-day follow-up. Propensity-score-matched 84 pairs of comparable patients. In the matched cohort (n = 168), tocilizumab was associated with fewer primary outcomes than the control group (hazard ratio (HR) = 0.49 (95% confidence interval (95%CI) = 0.3–0.81), p-value = 0.005). These results were similar in the overall cohort (n = 246), with Cox multivariable analysis yielding a protective association between tocilizumab and primary outcome (adjusted HR = 0.26 (95%CI = 0.135–0.51, p = 0.0001), confirmed by inverse probability score weighting (IPSW) analysis (p < 0.0001). Analyses on mortality only, with 28 days of follow-up, yielded similar results. In this study, tocilizumab 400 mg in a single-dose was associated with improved survival without mechanical ventilation in patients with severe COVID-19. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Figure 1
Survival curves, regarding the primary outcome with a 28-day follow-up, comparing tocilizumab and a control group. In the matched cohort (n = 168), tocilizumab was associated with fewer events (hazard ratio (HR) = 0.49 (95% confidence interval (95%CI) = 0.30–0.81), p = 0.005). In the overall cohort (n = 246), Cox multivariable survival analysis found tocilizumab to be independently associated with a lower incidence of the primary outcome (adjusted HR = 0.34 (95%CI = 0.22–0.52), p < 0.0001). Inverse probability score-weighted analysis yielded similar results (p < 0.0001). Full article ">Figure 2
Survival curves, regarding mortality with a 28-day follow-up, comparing tocilizumab and a control group. In the matched cohort (n = 168), tocilizumab was associated with fewer deaths (hazard ratio = 0.42 (95%CI = 0.22–0.82), p = 0.008). In the overall cohort (n = 246), Cox multivariable analysis yielded an independent protective association between tocilizumab and mortality (adjusted HR = 0.29 (95%CI = 0.17–0.53), p < 0.0001). Inverse probability score-weighted analysis yielded similar results (p < 0.0001). Full article ">Figure 3
Survival curves, regarding the primary outcome with a 28-day follow-up, comparing tocilizumab and a control group, after excluding patients who presented outcomes in the first 48 h after inclusion. Cox multivariable analysis yielded a protective association between tocilizumab and the primary outcome (adj.HR = 0.40 (95%CI = 0.23–0.70), p = 0.001). Full article ">Figure 4
Survival curves, regarding mortality with a 28-day follow-up, comparing tocilizumab and the control group, after excluding patients who presented outcomes in the first 48 h after inclusion. Treatment by tocilizumab was found to be protectively associated with mortality (adj.HR = 0.36 (95%CI = 0.18–0.70) p = 0.003). Full article ">

14 pages, 1460 KiB

Open AccessArticle

Squalene Emulsion Manufacturing Process Scale-Up for Enhanced Global Pandemic Response

by Tony Phan, Christian Devine, Erik D. Laursen, Adrian Simpson, Aaron Kahn, Amit P. Khandhar, Steven Mesite, Brad Besse, Ken J. Mabery, Elizabeth I. Flanagan and Christopher B. Fox

Pharmaceuticals 2020, 13(8), 168; https://doi.org/10.3390/ph13080168 - 28 Jul 2020

Cited by 6 | Viewed by 7833

Abstract

Squalene emulsions are among the most widely employed vaccine adjuvant formulations. Among the demonstrated benefits of squalene emulsions is the ability to enable vaccine antigen dose sparing, an important consideration for pandemic response. In order to increase pandemic response capabilities, it is desirable [...] Read more.

Squalene emulsions are among the most widely employed vaccine adjuvant formulations. Among the demonstrated benefits of squalene emulsions is the ability to enable vaccine antigen dose sparing, an important consideration for pandemic response. In order to increase pandemic response capabilities, it is desirable to scale up adjuvant manufacturing processes. We describe innovative process enhancements that enabled the scale-up of bulk stable squalene emulsion (SE) manufacturing capacity from a 3000- to 5,000,000-dose batch size. Manufacture of concentrated bulk along with the accompanying viscosity change in the continuous phase resulted in a ≥25-fold process efficiency enhancement. Process streamlining and implementation of single-use biocontainers resulted in reduced space requirements, fewer unit operations, and minimization of cleaning requirements. Emulsion physicochemical characteristics were measured by dynamic light scattering, laser diffraction, and HPLC with charged aerosol detection. The newly developed full-scale process was demonstrated by producing two 5,000,000-dose batches of bulk concentrated SE. A scale-up of adjuvant manufacturing capacity through process innovation enables more efficient production capabilities for pandemic response. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Small-scale (≤1 L) process flow diagram for production of SE. Full article ">Figure 2
Emulsion droplet diameter (a) and polydispersity index (b) as a function of dispersed phase volume percent and number of Microfluidizer processing passes. Droplet diameter was measured within one month following manufacture. Error bars represent standard deviation of three measurements from one batch of emulsion produced at each dispersed phase content level. Note that the specific volume of DMPC was not accounted for, thus the concentration of DMPC and other aqueous phase excipients may have varied between ~1–13% from the target concentrations [<a href="#B13-pharmaceuticals-13-00168" class="html-bibr">13</a>]. Full article ">Figure 3
Droplet diameter (a) and polydispersity index (b) as a function of number of passes through the Microfluidizer processor in 30% v/v oil-in-water emulsion batches produced at the pilot scale (10 L) using the one-pot approach or the traditional separate phases approach as indicated. Full article ">Figure 4
Droplet diameter (a) and polydispersity index (b) as a function of number of passes through the indicated Microfluidizer processor model. Error bars represent the standard deviation from three measurements from the same aliquot. The M-7250-30 data are taken from <a href="#pharmaceuticals-13-00168-f003" class="html-fig">Figure 3</a>. Batch size produced for each model was 10 L (M7250-30), 300 mL (M110-EH), 200 mL (M110P), and 100 mL (LM20). Typical representative flow rate for each Microfluidizer model when processing at 30,000 psi is 2.8 L/min (M7250-30), 400 mL/min (M110-EH), 110 mL/min (M110P), and 80 mL/min (LM20). Prior to Microfluidizer processing, high shear mixing conditions and equipment varied somewhat for the different batches although minimal impact is anticipated from these differences. Full article ">Figure 5
Physical stability of the two large-scale batches from <a href="#pharmaceuticals-13-00168-t007" class="html-table">Table 7</a> when stored at 2–8 °C. Droplet diameter (a) and polydispersity index (b) remain stable for a minimum of 12–18 months. Error bars represent the standard deviation of 8–9 measurements. Full article ">Figure 6
Large-scale (200 L) process flow diagram for one-pot production of SE. Full article ">

13 pages, 1520 KiB

Open AccessCommunication

Preliminary Virtual Screening Studies to Identify GRP78 Inhibitors Which May Interfere with SARS-CoV-2 Infection

by Andreia Palmeira, Emília Sousa, Aylin Köseler, Ramazan Sabirli, Tarık Gören, İbrahim Türkçüer, Özgür Kurt, Madalena M. Pinto and M. Helena Vasconcelos

Pharmaceuticals 2020, 13(6), 132; https://doi.org/10.3390/ph13060132 - 25 Jun 2020

Cited by 52 | Viewed by 6797 | Correction

Abstract

SARS-CoV-2 Spike protein was predicted by molecular docking to bind the host cell surface GRP78, which was suggested as a putative good molecular target to inhibit Covid-19. We aimed to confirm that GRP78 gene expression was increased in blood of SARS-CoV-2 (+) versus [...] Read more.

SARS-CoV-2 Spike protein was predicted by molecular docking to bind the host cell surface GRP78, which was suggested as a putative good molecular target to inhibit Covid-19. We aimed to confirm that GRP78 gene expression was increased in blood of SARS-CoV-2 (+) versus SARS-CoV-2 (−) pneumonia patients. In addition, we aimed to identify drugs that could be repurposed to inhibit GRP78, thus with potential anti-SARS-CoV-2 activity. Gene expression studies were performed in 10 SARS-CoV-2 (−) and 24 SARS-CoV-2 (+) pneumonia patients. A structure-based virtual screen was performed with 10,761 small molecules retrieved from DrugBank, using the GRP78 nucleotide binding domain and substrate binding domain as molecular targets. Results indicated that GRP78 mRNA levels were approximately four times higher in the blood of SARS-CoV-2 (+) versus SARS-CoV-2 (−) pneumonia patients, further suggesting that GRP78 might be a good molecular target to treat Covid-19. In addition, a total of 409 compounds were identified with potential as GRP78 inhibitors. In conclusion, we found preliminary evidence that further proposes GRP78 as a possible molecular target to treat Covid-19 and that many clinically approved drugs bind GRP78 as an off-target effect. We suggest that further work should be urgently carried out to confirm if GRP78 is indeed a good molecular target and if some of those drugs have potential to be repurposed for SARS-CoV-2 antiviral activity. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Docking studies on GRP78 nucleotide binding domain (NBD). (A) Three-dimensional (3D) surface structure of GRP78 (pdb code 5e84) showing its nucleotide-binding domains (yellow circle). (B) Crystallographic ATP, and (C) known drug Ponatinib, in 3D representation (left image) and respective two-dimensional (2D) interaction scheme (right image). The polar and non-polar amino-acids are shown in pink and green circles; hydrogen bonding is indicated by dotted arrows; with dotted lines represent arene-hydrogen interactions; proximity contour are dotted lines surrounding the ligand, indicating the shape of the binding site and available space to the more outward-facing parts of the ligand; blue shadows in some amino acids indicate the receptor exposure differences by the size and intensity of the quoits disks. The directions of the shadows indicate the directions of the amino acids toward the ligands. The blue clouds around the ligand atoms indicate the solvent exposure. Full article ">Figure 2
Docking studies on GRP78 SBD. (A) 3D surface structure of GRP78 (pdb code 5e84) (grey surface) bound to SARS-CoV-2 (red transparent surface); the C480–C488 section of the whole SARS-CoV-2 spike (pdb code 6m0j) was aligned with the docked C480–C488 in order to allow an easier visualization of the position of the spike towards the GRP78 target. Known peptide Zilucoplan is shown in sticks for exemplification. (B) SARS-CoV-2 region IV, and (C) known drug Zilucoplan bound to GRP78 SBD in 3D representation (left image, with arrows representing the direction of the peptidic strand from the linear—I, II, III—to the macrocyclic segment—IV) and respective 2D interaction scheme (right image). The polar and non-polar amino-acids are shown in pink and green circles; hydrogen bonding is indicated by dotted arrows; proximity contour are dotted lines surrounding the ligand, indicating the shape of the binding site and available space to the more outward-facing parts of the ligand; blue shadows in some amino acids indicate the receptor exposure differences by the size and intensity of the quoits disks. The directions of the shadows indicate the directions of the amino acids toward the ligands. The blue clouds around the ligand atoms indicate the solvent exposure. Full article ">

10 pages, 2523 KiB

Open AccessArticle

Excess Ascorbate is a Chemical Stress Agent against Proteins and Cells

by Maria Lehene, Eva Fischer-Fodor, Florina Scurtu, Niculina D. Hădade, Emese Gal, Augustin C. Mot, Alina Matei and Radu Silaghi-Dumitrescu

Pharmaceuticals 2020, 13(6), 107; https://doi.org/10.3390/ph13060107 - 27 May 2020

Cited by 7 | Viewed by 5076

Abstract

Excess ascorbate (as expected in intravenous treatment proposed for COVID-19 management, for example) oxidizes and/or degrades hemoglobin and albumin, as evidenced by UV-vis spectroscopy, gel electrophoresis, and mass spectrometry. It also degrades hemoglobin in intact blood or in isolated erythrocytes. The survival rates [...] Read more.

Excess ascorbate (as expected in intravenous treatment proposed for COVID-19 management, for example) oxidizes and/or degrades hemoglobin and albumin, as evidenced by UV-vis spectroscopy, gel electrophoresis, and mass spectrometry. It also degrades hemoglobin in intact blood or in isolated erythrocytes. The survival rates and metabolic activities of several leukocyte subsets implicated in the antiviral cellular immune response are also affected. Excess ascorbate is thus an unselective biological stress agent. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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UV-vis spectra of hemoglobin (a) panel: ferric; (b) panel: ferrous oxy with varying concentrations of ascorbate. Conditions: 5 µM met-Hb and 7 µM oxy-bovine-Hb, room temperature, 50 mM phosphate, pH 7. Full article ">Figure 2
UV-vis spectra of erythrocytes exposed to ascorbate. Conditions: Bovine erythrocytes re-suspended in phosphate buffer saline (pH 7.4, PBS), were incubated at 37 °C. Aliquots of the samples were retrieved at indicated times and diluted with PBS in the UV-vis cuvettes for spectral measurements. Experiments were performed in triplicate. Full article ">Figure 3
UV-vis spectra of whole blood exposed to ascorbate at (a) 4 h and (b) 24 h after mixing. Conditions: Bovine blood was incubated at 37 °C. Aliquots of the samples were retrieved at indicated times and diluted with PBS in the UV-vis cuvettes for spectral measurements. Experiments were performed in triplicate. Full article ">Figure 4
SDS-PAGE (12%) of hemoglobin and albumin exposed to ascorbate. Conditions: 50 µM Hb and 20 µM bovine serum albumin (BSA) were incubated at room temperature for the indicated times with or without 10 mM ascorbate in 50 mM phosphate, pH 7. The samples from left to right are: 1. Hb t0 (no incubation); 2. Hb 4 h; 3.Hb + ascorbate t0; 4. Hb + ascorbate 4 h; 5. Hb + ascorbate 24 h; 6.BSA t0; 7.BSA 4 h; 8.BSA + ascorbate t0; 9.BSA + ascorbate 4 h; 10.BSA + ascorbate 24 h. Full article ">Figure 5
Survival rates for human cell cultures exposed to indicated concentrations of ascorbate for 24 h. Full article ">Figure 6
Peripheral blood mononuclear cells (PBMC) and PBMC subsets, i.e., CD4+, CD8+, CD14+, CD19+, CD45RA+, and CD69+, metabolic activities expressed as reduction rates of resazurin dye. For all cell groups, ANOVA p values < 0.000, except the CD8+ subset, for which p < 0.005. Using the t-test for independent samples, the lowest ascorbic acid concentrations that significantly differed from the control was 10 mM for PMBC (p < 0.01), 25 mM for CD4+ (p < 0.000), 10 mM for CD8+ (p < 0.05), 25 mM for CD14+ (p < 0.001), 10 mM for CD19+ (p < 0.001), 10 mM for CD45RA+ (p < 0.005), and 25 mM for CD69+ (p < 0.001). Full article ">

Review

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18 pages, 1067 KiB

Open AccessReview

Repurposing Potential of the Antiparasitic Agent Ivermectin for the Treatment and/or Prophylaxis of COVID-19

by Hoda Awad, Basmala Hassan, Sara Dweek, Yasmeen Aboelata, Mutasem Rawas-Qalaji and Iman Saad Ahmed

Pharmaceuticals 2022, 15(9), 1068; https://doi.org/10.3390/ph15091068 - 27 Aug 2022

Cited by 3 | Viewed by 7197 | Correction

Abstract

Due to the rapid, vast, and emerging global spread of the Coronavirus Disease 2019 (COVID-19) pandemic, many drugs were quickly repurposed in a desperate attempt to unveil a miracle drug. Ivermectin (IVM), an antiparasitic macrocyclic lactone, was tested and confirmed for its in [...] Read more.

Due to the rapid, vast, and emerging global spread of the Coronavirus Disease 2019 (COVID-19) pandemic, many drugs were quickly repurposed in a desperate attempt to unveil a miracle drug. Ivermectin (IVM), an antiparasitic macrocyclic lactone, was tested and confirmed for its in vitro antiviral activity against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in early 2020. Along with its potential antiviral activity, the affordability and availability of IVM resulted in a wide public interest. Across the world, trials have put IVM to test for both the treatment and prophylaxis of COVID-19, as well as its potential role in combination therapy. Additionally, the targeted delivery of IVM was studied in animals and COVID-19 patients. Through this conducted literature review, the potential value and effectiveness of the repurposed antiparasitic agent in the ongoing global emergency were summarized. The reviewed trials suggested a value of IVM as a treatment in mild COVID-19 cases, though the benefit was not extensive. On the other hand, IVM efficacy as a prophylactic agent was more evident and widely reported. In the most recent trials, novel nasal formulations of IVM were explored with the hope of an improved optimized effect. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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17 pages, 1429 KiB

Open AccessReview

Immunosuppressant Therapies in COVID-19: Is the TNF Axis an Alternative?

by Yadira Palacios and Leslie Chavez-Galan

Pharmaceuticals 2022, 15(5), 616; https://doi.org/10.3390/ph15050616 - 17 May 2022

Cited by 9 | Viewed by 6027

Abstract

The study of cytokine storm in COVID-19 has been having different edges in accordance with the knowledge of the disease. Various cytokines have been the focus, especially to define specific treatments; however, there are no conclusive results that fully support any of the [...] Read more.

The study of cytokine storm in COVID-19 has been having different edges in accordance with the knowledge of the disease. Various cytokines have been the focus, especially to define specific treatments; however, there are no conclusive results that fully support any of the options proposed for emergency treatment. One of the cytokines that requires a more exhaustive review is the tumor necrosis factor (TNF) and its receptors (TNFRs) as increased values of soluble formats for both TNFR1 and TNFR2 have been identified. TNF is a versatile cytokine with different impacts at the cellular level depending on the action form (transmembrane or soluble) and the receptor to which it is associated. In that sense, the triggered mechanisms can be diversified. Furthermore, there is the possibility of the joint action provided by synergism between one or more cytokines with TNF, where the detonation of combined cellular processes has been suggested. This review aims to discuss some roles of TNF and its receptors in the pro-inflammatory stage of COVID-19, understand its ways of action, and let to reposition this cytokine or some of its receptors as therapeutic targets. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Graphical abstract

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Activation of the immune response during the SARS-CoV-2 infection establishment. (A) Alveolar epithelial cell type II express high levels of ACE2 used as the primary cell receptor for SARS-CoV-2 through the S protein activation by the protease TMPRSS2. PAMPs and DAMPs are the first signals to promote innate immune activation. The chemokines CXCL8 and CCL2 promote cell mobilization (CXCL refers to the C-X-C motif chemokine ligand number, where “C” is a cysteine, and “X” represents any amino acid; CCL refers to the C-C motif chemokine ligand number, where “C” is a cysteine). (B) In severe COVID-19, there are hyperinflammatory macrophages that promote the release of cytokines and chemokines like IL-6, IL-8, TNF, IFN-γ, IL-1β, CXCL10, CCL8, CCL20, CXCL2, CXCL3, CCL3, CXCL3, and CCL4. (C) PAMPs and DAMPs also favor inflammasome activation. The NLRP3 formation is the central platform for caspase-1 activation and the proteolytic maturation of IL-1β and IL-18. (D) IL-17 and IL-18 activate neutrophils in the alveolar space evolving into NETosis, which promotes more inflammation. (E) Together, all pro-inflammatory signals induce the T cell recruitment, which produces TNF and IFN-γ, and it impacts endothelial and dendritic cells. (Figure created with BioRender.com, adapted from “Cytokine storm template” by BioRender.com. Obtained with <a href="https://app.biorender.com/biorender-templates" target="_blank">https://app.biorender.com/biorender-templates</a>, accessed on 21 April 2022). Full article ">Figure 2
TNF inhibitors approved for therapeutical use. Currently, there are five monoclonal antibodies used in anti-TNF therapy; each one has a specific structure and origin. However, all of them neutralize both transmembrane (tm) and soluble (s) TNF. (Figure created with <a href="http://BioRender.com" target="_blank">BioRender.com</a>, accessed on 21 April 2022). Full article ">

13 pages, 335 KiB

Open AccessReview

COVID-19 and Seasonal Influenza Vaccination: Cross-Protection, Co-Administration, Combination Vaccines, and Hesitancy

by Alexander Domnich, Andrea Orsi, Carlo-Simone Trombetta, Giulia Guarona, Donatella Panatto and Giancarlo Icardi

Pharmaceuticals 2022, 15(3), 322; https://doi.org/10.3390/ph15030322 - 8 Mar 2022

Cited by 37 | Viewed by 6633

Abstract

SARS-CoV-2 and influenza are the main respiratory viruses for which effective vaccines are currently available. Strategies in which COVID-19 and influenza vaccines are administered simultaneously or combined into a single preparation are advantageous and may increase vaccination uptake. Here, we comprehensively review the [...] Read more.

SARS-CoV-2 and influenza are the main respiratory viruses for which effective vaccines are currently available. Strategies in which COVID-19 and influenza vaccines are administered simultaneously or combined into a single preparation are advantageous and may increase vaccination uptake. Here, we comprehensively review the available evidence on COVID-19/influenza vaccine co-administration and combination vaccine candidates from the standpoints of safety, immunogenicity, efficacy, policy and public acceptance. While several observational studies have shown that the trained immunity induced by influenza vaccines can protect against some COVID-19-related endpoints, it is not yet understood whether co-administration or combination vaccines can exert additive effects on relevant outcomes. In randomized controlled trials, co-administration has proved safe, with a reactogenicity profile similar to that of either vaccine administered alone. From the immunogenicity standpoint, the immune response towards four influenza strains and the SARS-CoV-2 spike protein in co-administration groups is generally non-inferior to that seen in groups receiving either vaccine alone. Several public health authorities have advocated co-administration. Different combination vaccine candidates are in (pre)-clinical development. The hesitancy towards vaccine co-administration or combination vaccines is a multifaceted phenomenon and may be higher than the acceptance of either vaccine administered separately. Public health implications are discussed. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

17 pages, 662 KiB

Open AccessEditor’s ChoiceReview

Monoclonal Antibodies against SARS-CoV-2: Current Scenario and Future Perspectives

by Eugenia Quiros-Roldan, Silvia Amadasi, Isabella Zanella, Melania Degli Antoni, Samuele Storti, Giorgio Tiecco and Francesco Castelli

Pharmaceuticals 2021, 14(12), 1272; https://doi.org/10.3390/ph14121272 - 6 Dec 2021

Cited by 25 | Viewed by 6094

Abstract

Monoclonal antibodies (mAbs) have been known since the 1970s. However, their therapeutic potential in the medical field has recently emerged, with the advancement of manufacturing techniques. Initially exploited mainly in the oncology field, mAbs have become increasingly relevant in Infectious Diseases. Numerous mAbs [...] Read more.

Monoclonal antibodies (mAbs) have been known since the 1970s. However, their therapeutic potential in the medical field has recently emerged, with the advancement of manufacturing techniques. Initially exploited mainly in the oncology field, mAbs have become increasingly relevant in Infectious Diseases. Numerous mAbs have been developed against SARS-CoV 2 and have proven their effectiveness, especially in the management of the mild-to-moderate disease. In this review, we describe the monoclonal antibodies currently authorized for the treatment of the coronavirus disease 19 (COVID-19) and offer an insight into the clinical trials that led to their approval. We discuss the mechanisms of action and methods of administration as well as the prophylactic and therapeutic labelled indications (both in outpatient and hospital settings). Furthermore, we address the critical issues regarding mAbs, focusing on their effectiveness against the variants of concern (VoC) and their role now that a large part of the population has been vaccinated. The purpose is to offer the clinician an up-to-date overview of a therapeutic tool that could prove decisive in treating patients at high risk of progression to severe disease. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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23 pages, 1324 KiB

Open AccessFeature PaperReview

Antirheumatic Drugs against COVID-19 from the Perspective of Rheumatologists

by Mai Kawazoe, Mari Kihara and Toshihiro Nanki

Pharmaceuticals 2021, 14(12), 1256; https://doi.org/10.3390/ph14121256 - 2 Dec 2021

Cited by 12 | Viewed by 5612

Abstract

Coronavirus disease 2019 (COVID-19) remains a global threat to humanity. Its pathogenesis and different phases of disease progression are being elucidated under the pandemic. Active viral replication activates various immune cells and produces large amounts of inflammatory cytokines, which leads to the cytokine [...] Read more.

Coronavirus disease 2019 (COVID-19) remains a global threat to humanity. Its pathogenesis and different phases of disease progression are being elucidated under the pandemic. Active viral replication activates various immune cells and produces large amounts of inflammatory cytokines, which leads to the cytokine storm, a major cause of patient death. Therefore, viral inhibition is expected to be the most effective early in the course of the disease, while immunosuppressive treatment may be useful in the later stages to prevent disease progression. Based on the pathophysiology of rheumatic diseases, various immunomodulatory and immunosuppressive drugs are used for the diseases. Due to their mechanism of action, the antirheumatic drugs, including hydroxychloroquine, chloroquine, colchicine, calcineurin inhibitors (e.g., cyclosporine A and tacrolimus), glucocorticoids, cytokines inhibitors, such as anti-tumor necrosis factor-α (e.g., infliximab), anti-interleukin (IL)-6 (e.g., tocilizumab, sarilumab, and siltuximab), anti-IL-1 (e.g., anakinra and canakinumab) and Janus kinase inhibitors (e.g., baricitinib and tofacitinib), cytotoxic T lymphocyte-associated antigen 4 blockade agents (e.g., abatacept), and phosphodiesterase 4 inhibitors (e.g., apremilast), have been tried as a treatment for COVID-19. In this review, we discuss the mechanisms of action and clinical impact of these agents in the management of COVID-19. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Figure 1
Classification of COVID-19 states and potential therapies. The figure illustrates the three escalating phases of COVID-19 progression with associated symptoms and potential phase-specific therapies. ARDS, acute respiratory distress syndrome; CTLA, cytotoxic T lymphocyte-associated antigen; IL, interleukin; JAK, Janus kinase; MOF, multiple organ failure; PDE, phosphodiesterase. Full article ">Figure 2
Overview of the mechanism of SARS-CoV-2 infection. The spike (S) protein on the surface of SARS-CoV-2 binds to ACE2 expressed on host cells, such as nasal and bronchial epithelial cells and pneumocytes. TMPRSS2 present on the host cell surface subsequently primes the S protein and promotes endocytosis-induced viral entry into the cell. The virus is carried into the endosome and uncoated by fusing with the endosome membrane, which releases its genomic RNA into the cytoplasm. Genomic RNA replication and protein synthesis occur in the ribosome, forming viral particles and releasing them extracellularly. Active viral replication activates various immune cells and produces large amounts of inflammatory cytokines. CQ and HCQ inhibits the glycosylation of ACE2, which may interfere with the binding of SARS-CoV-2 to the cell receptor. CQ and HCQ decrease acidity in endosomes and inhibit the fusion of SARS-CoV-2 to host cell membranes, and also interfere with TLR signaling by changing local pH. Colchicine may affect clathrin-mediated endocytosis. BAR binds AAK1 and GAK, the identified regulators of endocytosis, and reduces viral entry. ACE2, angiotensin-converting enzyme II; AAK1, AP2-associated protein kinase 1; BAR, baricitinib; CQ, Chloroquine; GAK, cyclin G-associated kinase; HCQ, hydroxychloroquine; TLR, Toll-like receptor; TMPRSS2, transmembrane protease serine 2. Full article ">Figure 3
Schematic representation of sites of action of antirheumatic drugs. Janus kinases (JAK1, JAK2, JAK3, and TYK2) are activated by extracellular stimuli, including cytokines, and phosphorylate downstream STAT proteins, which translocate to the nucleus and activate target genes to produce inflammatory cytokines. IFN, interferon; IL, interleukin; JAK, Janus kinase; P, phosphoric acid; STAT, signal transducer and activator of transcription; TNF, tumor necrosis factor; TYK, tyrosine kinase. Full article ">

13 pages, 312 KiB

Open AccessReview

The Impact of SARS-CoV-2 Infection, and Application of Immunosuppressive Agents in Kidney Transplant Recipients Suffering from COVID-19

by Horng-Ta Tseng, Xiang-Chi Wu, Chun-Yao Huang, Chun-Ming Shih, Yi-Wen Lin and Feng-Yen Lin

Pharmaceuticals 2021, 14(10), 1054; https://doi.org/10.3390/ph14101054 - 17 Oct 2021

Cited by 4 | Viewed by 2527

Abstract

In December 2019, the COVID-19 pandemic began to ravage the world quickly, causing unprecedented losses in human life and the economy. A statistical study revealed that the proportion of solid organ transplant (SOT) recipients with severe symptoms and deaths after being infected by [...] Read more.

In December 2019, the COVID-19 pandemic began to ravage the world quickly, causing unprecedented losses in human life and the economy. A statistical study revealed that the proportion of solid organ transplant (SOT) recipients with severe symptoms and deaths after being infected by SARS-CoV-2 is considerably higher than that of non-SOT recipients, and the prognosis is relatively poor. In addition, the clinical manifestation of SOT recipients suffering from COVID-19 is different from that of general COVID-19 patients. Acute kidney injury (AKI) is a common complication in COVID-19 patients, and it is likely more common among SOT recipients infected with SARS-CoV-2. Clinical experts consider that SOT recipients have long-term treatment with immunosuppressants, and the comorbidities are driven by a high rate of severe symptoms and mortality. Orthotopic kidney allograft transplantation is an effective treatment for patients suffering from end-stage kidney disease/kidney failure through which they can easily extend their life. Indeed, kidney transplant recipients have suffered significant damage during this pandemic. To effectively reduce the severity of symptoms and mortality of kidney transplant recipients suffering from COVID-19, precise application of various drugs, particularly immunosuppressants, is necessary. Therefore, herein, we will collate the current clinical experience of treating COVID-19 infection in kidney transplant recipients and discuss the adjustment of patients using immunosuppressive agents in the face of COVID-19. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

16 pages, 750 KiB

Open AccessReview

Pharmacology and Adverse Events of Emergency-Use Authorized Medication in Moderate to Severe COVID-19

by Jen-Yu Hsu, Yan-Chiao Mao, Po-Yu Liu and Kuo-Lung Lai

Pharmaceuticals 2021, 14(10), 955; https://doi.org/10.3390/ph14100955 - 23 Sep 2021

Cited by 15 | Viewed by 3059

Abstract

Some effective drugs have been approved or issued an Emergency Use Authorization for the treatment of COVID-19 in hospitalized patients, but post-market surveillance is warranted to monitor adverse events. We reviewed clinical trials and case reports in patients with moderate-to-severe COVID-19 infection who [...] Read more.

Some effective drugs have been approved or issued an Emergency Use Authorization for the treatment of COVID-19 in hospitalized patients, but post-market surveillance is warranted to monitor adverse events. We reviewed clinical trials and case reports in patients with moderate-to-severe COVID-19 infection who received remdesivir, baricitinib, tocilizumab, or sarilumab. The drug-specific pharmacokinetics, toxicity, and drug interactions are summarized in this study. Remdesivir and baricitinib are small-molecule drugs that are mainly metabolized by the kidneys, while tocilizumab and sarilumab are monoclonal antibody drugs with metabolic pathways that are currently not fully understood. The most common adverse events of these drugs are alterations in liver function, but serious adverse events have rarely been attributed to them. Only a few studies have reported that remdesivir might be cardiotoxic and that baricitinib might cause thromboembolism. Biological agents such as baricitinib, tocilizumab, and sarilumab could inhibit the pathway of inflammatory processes, leading to immune dysregulation, so the risk of secondary infection should be assessed before prescribing. Further recognition of the pathogenic mechanism and risk factors of adverse events is essential for optimizing treatment strategies. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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18 pages, 1700 KiB

Open AccessReview

Neuropsychiatric Disorders and COVID-19: What We Know So Far

by Fernanda Majolo, Guilherme Liberato da Silva, Lucas Vieira, Cetin Anli, Luís Fernando Saraiva Macedo Timmers, Stefan Laufer and Márcia Inês Goettert

Pharmaceuticals 2021, 14(9), 933; https://doi.org/10.3390/ph14090933 - 17 Sep 2021

Cited by 15 | Viewed by 4299

Abstract

SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus-2) affects the central nervous system (CNS), which is shown in a significant number of patients with neurological events. In this study, an updated literature review was carried out regarding neurological disorders in COVID-19. Neurological symptoms are more [...] Read more.

SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus-2) affects the central nervous system (CNS), which is shown in a significant number of patients with neurological events. In this study, an updated literature review was carried out regarding neurological disorders in COVID-19. Neurological symptoms are more common in patients with severe infection according to their respiratory status and divided into three categories: (1) CNS manifestations; (2) cranial and peripheral nervous system manifestations; and (3) skeletal muscle injury manifestations. Patients with pre-existing cerebrovascular disease are at a higher risk of admission to the intensive care unit (ICU) and mortality. The neurological manifestations associated with COVID-19 are of great importance, but when life-threatening abnormal vital signs occur in severely ill COVID-19 patients, neurological problems are usually not considered. It is crucial to search for new treatments for brain damage, as well as for alternative therapies that recover the damaged brain and reduce the inflammatory response and its consequences for other organs. In addition, there is a need to diagnose these manifestations as early as possible to limit long-term consequences. Therefore, much research is needed to explain the involvement of SARS-CoV-2 causing these neurological symptoms because scientists know zero about it. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Pathways through which viruses can infect the peripheral nervous system (PNS) or central nervous system (CNS): (1) nerve endings were accessed in the tissues by the infection using axonal transport machinery to gain access to the CNS; or (2) by the infected cells in a circulatory system, which carries the infection through the blood–brain barrier into the CNS. Adapted from Yachou et al., 2020 [<a href="#B98-pharmaceuticals-14-00933" class="html-bibr">98</a>]. Full article ">Figure 2
SARS-CoV-2 infecting cells in lung tissue through the interaction with angiotensin-converting enzyme 2 (ACE2), promoting the generation of pro-inflammatory cytokines (cytokine storm). This process attracts “defense” cells such as macrophages, monocytes, and T cells to the site of infection, triggering the inflammation process. The result of this inflammation is lung tissue damage. Adapted from Tay et al., 2020 [<a href="#B102-pharmaceuticals-14-00933" class="html-bibr">102</a>]. Full article ">Figure 3
Microglia and astrocytes are activated by ATP expressed during the release of pro-inflammatory cytokines. NMDA receptors expressed by Glutamate activation allow the Ca2+-dependent exocytosis of ATP, thus releasing more Glutamate. A massive release of these neurotransmitters increases cell death and excitotoxicity, since postsynaptic neuron increased the Ca2+-calmodulin (CaM) complex formation and consequent nNOS activation. Neurotoxicity is mediated by NO production, which interacts with the iron–sulfur centers in the mitochondrial electron transport chain, and produces reactive oxygen species (ROS), impairing cellular energy production. Still, a reaction of O2− (superoxide ion) and NO forms peroxynitrite (ONOO−) and peroxynitrous acid (ONOOH). Such formation leads to oxidative stress, which includes DNA damage, lipid peroxidation, tyrosine nitration, and excess S-nitrosylation, causing neuronal impairment and/or death. The activation of caspase-1 is mediated by the NLRP3 inflammasome by the cleavage of pro-IL-1β and pro-IL-18 in IL-1β and IL-18, respectively. The mature forms of cytokines are secreted worsening the neuroinflammatory process established. Then, the neuroinflammatory process established can be worsened by these mature cytokines. Adapted from Ribeiro et al., 2021 [<a href="#B105-pharmaceuticals-14-00933" class="html-bibr">105</a>]. Full article ">

13 pages, 1611 KiB

Open AccessReview

Prevalence and Significance of Hypermetabolic Lymph Nodes Detected by 2-[¹⁸F]FDG PET/CT after COVID-19 Vaccination: A Systematic Review and a Meta-Analysis

by Giorgio Treglia, Marco Cuzzocrea, Luca Giovanella, Luigia Elzi and Barbara Muoio

Pharmaceuticals 2021, 14(8), 762; https://doi.org/10.3390/ph14080762 - 3 Aug 2021

Cited by 28 | Viewed by 6248

Abstract

Recently, several articles reported incidental findings at 2-[¹⁸F]FDG PET/CT in patients who have received COVID-19 vaccinations, including hypermetabolic axillary lymph nodes (HALNs) ipsilateral to the COVID-19 vaccine injection site which may cause diagnostic dilemmas. The aim of our work was to [...] Read more.

Recently, several articles reported incidental findings at 2-[¹⁸F]FDG PET/CT in patients who have received COVID-19 vaccinations, including hypermetabolic axillary lymph nodes (HALNs) ipsilateral to the COVID-19 vaccine injection site which may cause diagnostic dilemmas. The aim of our work was to calculate the prevalence of this finding. A comprehensive computer literature search of PubMed/MEDLINE, Embase, and Cochrane library databases was performed to identify recently published articles that investigated the prevalence of HALNs detected by 2-[¹⁸F]FDG PET/CT after COVID-19 vaccination. Pooled prevalence of this finding was calculated through a meta-analytic approach. Nine recently published articles including 2354 patients undergoing 2-[¹⁸F]FDG PET/CT after recent COVID-19 vaccination have been included in the systematic review. Overall, HALNs ipsilateral to the vaccine injection site were frequent findings mainly due to vaccine-related immune response in most of the cases. The pooled prevalence of HALNs after COVID-19 vaccination was 37% (95% confidence interval: 27–47%) but with significant heterogeneity among the included studies. Physicians must be aware and recognize the significant frequency of HALNs at 2-[¹⁸F]FDG PET/CT related to immune response to vaccine injection. Larger studies are needed to confirm the findings of this systematic review and meta-analysis. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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13 pages, 1481 KiB

Open AccessReview

Current Status of Baricitinib as a Repurposed Therapy for COVID-19

by Maha Saber-Ayad, Sarah Hammoudeh, Eman Abu-Gharbieh, Rifat Hamoudi, Hamadeh Tarazi, Taleb H. Al-Tel and Qutayba Hamid

Pharmaceuticals 2021, 14(7), 680; https://doi.org/10.3390/ph14070680 - 15 Jul 2021

Cited by 22 | Viewed by 6432

Abstract

The emergence of the COVID-19 pandemic has mandated the instant (re)search for potential drug candidates. In response to the unprecedented situation, it was recognized early that repurposing of available drugs in the market could timely save lives, by skipping the lengthy phases of [...] Read more.

The emergence of the COVID-19 pandemic has mandated the instant (re)search for potential drug candidates. In response to the unprecedented situation, it was recognized early that repurposing of available drugs in the market could timely save lives, by skipping the lengthy phases of preclinical and initial safety studies. BenevolentAI’s large knowledge graph repository of structured medical information suggested baricitinib, a Janus-associated kinase inhibitor, as a potential repurposed medicine with a dual mechanism; hindering SARS-CoV2 entry and combatting the cytokine storm; the leading cause of mortality in COVID-19. However, the recently-published Adaptive COVID-19 Treatment Trial-2 (ACTT-2) positioned baricitinib only in combination with remdesivir for treatment of a specific category of COVID-19 patients, whereas the drug is not recommended to be used alone except in clinical trials. The increased pace of data output in all life sciences fields has changed our understanding of data processing and manipulation. For the purpose of drug design, development, or repurposing, the integration of different disciplines of life sciences is highly recommended to achieve the ultimate benefit of using new technologies to mine BIG data, however, the final say remains to be concluded after the drug is used in clinical practice. This review demonstrates different bioinformatics, chemical, pharmacological, and clinical aspects of baricitinib to highlight the repurposing journey of the drug and evaluates its placement in the current guidelines for COVID-19 treatment. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Flow chart of database searching. Full article ">Figure 2
Dual mechanism of action of baracitinib; (A) to inhibit clathrin-mediated endocytosis of the SARS-CoV2, and (B) to inhibit the JAK-mediate release of pro-inflammatory cytokines. Full article ">Figure 3
The effect of baracitinib treatment on ACE2 and TMPRSS2 expression (generated from dataset GSE61552). The dataset compiled the expression profile of systemically barictinib-treated and ruxolitinib-treated C3H/HeJ grafted model of alopecia areata. The analysis revealed a significant reduction of ACE2 and TMPRSS2 expression in the baricitinib-treated samples, compared to untreated control ones. ACE2 = angiotensin-converting enzyme 2, TMPRSS2: transmembrane protease serine 2. ** p-value < 0.01. Full article ">Figure 4
Structures of baricitinib, ruxolitinib, and fedratinib. Full article ">Figure 5
(A) An overlay of ruxolitinib (Purple) on baricitinib (Green); electronic similarity percentage = 82.5%. (B) An overlay of fedratinib (Gray) on ruxolitinib (Purple); electronic similarity percentage = 54.0%. (C) An overlay of fedratinib (Gray) on baricitinib (Green); electronic similarity percentage = 49.1%. Similarity and alignments were measured based on molecular fields descriptors generated by Cresset’s FieldAlign Software (version 1.0.2), [<a href="#B37-pharmaceuticals-14-00680" class="html-bibr">37</a>]. Full article ">

22 pages, 4263 KiB

Open AccessReview

Therapeutic Potential of Glycosyl Flavonoids as Anti-Coronaviral Agents

by Patrícia I. C. Godinho, Raquel G. Soengas and Vera L. M. Silva

Pharmaceuticals 2021, 14(6), 546; https://doi.org/10.3390/ph14060546 - 7 Jun 2021

Cited by 25 | Viewed by 4909

Abstract

The COVID-19 pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has spread all over the world, creating a devastating socio-economic impact. Even though protective vaccines are starting to be administered, an effective antiviral agent for the prevention and treatment of COVID-19 [...] Read more.

The COVID-19 pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has spread all over the world, creating a devastating socio-economic impact. Even though protective vaccines are starting to be administered, an effective antiviral agent for the prevention and treatment of COVID-19 is not available yet. Moreover, since new and deadly CoVs can emerge at any time with the potential of becoming pandemics, the development of therapeutic agents against potentially deadly CoVs is a research area of much current interest. In the search for anti-coronaviral drugs, researchers soon turned their heads towards glycosylated flavonoids. Glycosyl flavonoids, widespread in the plant kingdom, have received a lot of attention due to their widely recognized antioxidant, anti-inflammatory, neuroprotective, anticarcinogenic, antidiabetic, antimicrobial, and antiviral properties together with their capacity to modulate key cellular functions. The wide range of biological activities displayed by glycosyl flavonoids, along with their low toxicity, make them ideal candidates for drug development. In this review, we examine and discuss the up-to-date developments on glycosyl flavonoids as evidence-based natural sources of antivirals against coronaviruses and their potential role in the management of COVID-19. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Full article ">Figure 1
Schematic diagram of the coronavirus virion. Full article ">Figure 2
Overview of lung pathology in patients with severe acute respiratory syndrome coronavirus 2. The black arrows show the pathways of lung lesions after inhalation and entry of SARS-CoV-2 into the respiratory tract. The red arrows show the increase of inflammation markers. Full article ">Figure 3
Cryo-EM structure of the SARS-CoV-2 spike glycoprotein. The closed state (left) and open state (right) of the spike glycoprotein (closed state PDB: 6VXX and open state PDB: 6VYB). Full article ">Figure 4
Anti-inflammatory and antioxidant glycosyl flavonoids. Full article ">Figure 5
Glycosyl flavonoids with α-glucosidase inhibitory activity. Full article ">Figure 6
Antiviral glycosyl flavonoids. Full article ">Figure 7
Anti-coronaviral glycosyl flavonoids. Full article ">

28 pages, 651 KiB

Open AccessFeature PaperEditor’s ChoiceReview

Clinical Management of COVID-19: A Review of Pharmacological Treatment Options

by Ashli M. Heustess, Melissa A. Allard, Dorothea K. Thompson and Pius S. Fasinu

Pharmaceuticals 2021, 14(6), 520; https://doi.org/10.3390/ph14060520 - 28 May 2021

Cited by 23 | Viewed by 8353

Abstract

Since the outbreak and subsequent declaration of COVID-19 as a global pandemic in March 2020, concerted efforts have been applied by the scientific community to curtail the spread of the disease and find a cure. While vaccines constitute a vital part of the [...] Read more.

Since the outbreak and subsequent declaration of COVID-19 as a global pandemic in March 2020, concerted efforts have been applied by the scientific community to curtail the spread of the disease and find a cure. While vaccines constitute a vital part of the public health strategy to reduce the burden of COVID-19, the management of this disease will continue to rely heavily on pharmacotherapy. This study aims to provide an updated review of pharmacological agents that have been developed and/or repurposed for the treatment of COVID-19. To this end, a comprehensive literature search was conducted using the PubMed, Google Scholar, and LitCovid databases. Relevant clinical studies on drugs used in the management of COVID-19 were identified and evaluated in terms of evidence of efficacy and safety. To date, the FDA has approved three therapies for the treatment of COVID-19 Emergency Use Authorization: convalescent plasma, remdesivir, and casirivimab/imdevimab (REGN-COV2). Drugs such as lopinavir/ritonavir, umifenovir, favipiravir, anakinra, chloroquine, hydroxychloroquine, tocilizumab, interferons, tissue plasminogen activator, intravenous immunoglobulins, and nafamosat have been used off-label with mixed therapeutic results. Adjunctive administration of corticosteroids is also very common. The clinical experience with these approved and repurposed drugs is limited, and data on efficacy for the new indication are not strong. Overall, the response of the global scientific community to the COVID-19 pandemic has been impressive, as evident from the volume of scientific literature elucidating the molecular biology and pathophysiology of SARS-CoV-2 and the approval of three new drugs for clinical management. Reviewed studies have shown mixed data on efficacy and safety of the currently utilized drugs. The lack of standard treatment for COVID-19 has made it difficult to interpret results from most of the published studies due to the risk of attribution error. The long-term effects of drugs can only be assessed after several years of clinical experience; therefore, the efficacy and safety of current COVID-19 therapeutics should continue to be rigorously monitored as part of post-marketing studies. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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19 pages, 1069 KiB

Open AccessReview

New Approaches and Repurposed Antiviral Drugs for the Treatment of the SARS-CoV-2 Infection

by Luana Vittoria Bauso, Chiara Imbesi, Gasparo Irene, Gabriella Calì and Alessandra Bitto

Pharmaceuticals 2021, 14(6), 503; https://doi.org/10.3390/ph14060503 - 25 May 2021

Cited by 9 | Viewed by 4420

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the virus that causes coronavirus disease 2019 (COVID-19). The outbreak of this coronavirus was first identified in Wuhan (Hubei, China) in December 2019, and it was declared as pandemic by the World Health Organization (WHO) [...] Read more.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the virus that causes coronavirus disease 2019 (COVID-19). The outbreak of this coronavirus was first identified in Wuhan (Hubei, China) in December 2019, and it was declared as pandemic by the World Health Organization (WHO) in March 2020. Today, several vaccines against SARS-CoV-2 have been approved, and some neutralizing monoclonal antibodies are being tested as therapeutic approaches for COVID-19 but, one of the key questions is whether both vaccines and monoclonal antibodies could be effective against infections by new SARS-CoV-2 variants. Nevertheless, there are currently more than 1000 ongoing clinical trials focusing on the use and effectiveness of antiviral drugs as a possible therapeutic treatment. Among the classes of antiviral drugs are included 3CL protein inhibitors, RNA synthesis inhibitors and other small molecule drugs which target the ability of SARS-COV-2 to interact with host cells. Considering the need to find specific treatment to prevent the emergent outbreak, the aim of this review is to explain how some repurposed antiviral drugs, indicated for the treatment of other viral infections, could be potential candidates for the treatment of COVID-19. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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16 pages, 1098 KiB

Open AccessReview

Tenofovir, Another Inexpensive, Well-Known and Widely Available Old Drug Repurposed for SARS-COV-2 Infection

by Isabella Zanella, Daniela Zizioli, Francesco Castelli and Eugenia Quiros-Roldan

Pharmaceuticals 2021, 14(5), 454; https://doi.org/10.3390/ph14050454 - 11 May 2021

Cited by 34 | Viewed by 5772 | Correction

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is spreading worldwide with different clinical manifestations. Age and comorbidities may explain severity in critical cases and people living with human immunodeficiency virus (HIV) might be at particularly high risk for severe progression. Nonetheless, current [...] Read more.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is spreading worldwide with different clinical manifestations. Age and comorbidities may explain severity in critical cases and people living with human immunodeficiency virus (HIV) might be at particularly high risk for severe progression. Nonetheless, current data, although sometimes contradictory, do not confirm higher morbidity, risk of more severe COVID-19 or higher mortality in HIV-infected people with complete access to antiretroviral therapy (ART). A possible protective role of ART has been hypothesized to explain these observations. Anti-viral drugs used to treat HIV infection have been repurposed for COVID-19 treatment; this is also based on previous studies on severe acute respiratory syndrome virus (SARS-CoV) and Middle East respiratory syndrome virus (MERS-CoV). Among them, lopinavir/ritonavir, an inhibitor of viral protease, was extensively used early in the pandemic but it was soon abandoned due to lack of effectiveness in clinical trials. However, remdesivir, a nucleotide analog that acts as reverse-transcriptase inhibitor, which was tested early during the pandemic because of its wide range of antiviral activity against several RNA viruses and its safety profile, is currently the only antiviral medication approved for COVID-19. Tenofovir, another nucleotide analog used extensively for HIV treatment and pre-exposure prophylaxis (PrEP), has also been hypothesized as effective in COVID-19. No data on tenofovir’s efficacy in coronavirus infections other than COVID-19 are currently available, although information relating to SARS-CoV-2 infection is starting to come out. Here, we review the currently available evidence on tenofovir’s efficacy against SARS-CoV-2. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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(a) SARS-CoV-2 is an enveloped positive-sense single-stranded RNA virus. In mature virions, structural proteins surround and interact with the viral genome (spike (S), envelope (E), membrane (M) and nucleocapsid (N) proteins). Virions enter target cells after specific binding of the receptor-binding domain (RBD) of the viral protein S to the cellular entry receptor, the angiotensin-converting enzyme 2 (ACE2). Viral uptake is promoted by the proteolytic activity of host factors, like the cell-surface transmembrane serine protease 2 (TMPRSS2), whose enzymatic activity is essential to permit the fusion with the cellular membrane and virus entry. Once entered target cells, virions are uncoated and viral RNA is released in the cytosol and immediately translated into two large polyproteins, pp1a and pp1ab, specified by two large open reading frames of the viral RNA, ORF1a and ORF1b. Both polyproteins contain the amino acid sequences of viral non-structural proteins (nsps). pp1a contains nsp1 to nsp11, while pp1ab contains nsp1 to nsp10 and nsp12 to nsp16. These polyproteins are co-translationally and post-translationally processed through proteolytic cleavage by two viral cysteine proteases residing in the nsp3 and nsp5 sequence, the papain-like protease (PLpro) and the main protease or 3C-like protease (Mpro/3CLpro), respectively. Released nsp1 inhibits host mRNA translation, while nsp2 to nsp16 are involved in the intracellular viral replication cycle; in particular, nsp12 is the RNA dependent RNA polymerase (RdRp) that with the cofactors nsp7 and nsp8 performs viral RNA synthesis for translation and synthesis of structural and accessory proteins and for the assembly of new virions. (b) SARS-CoV-2 genome and protein organization. Full article ">Figure 2
Chemical structures of tenofovir, tenofovir disoproxil fumarate (TDF) and tenofovir alafenamide (TAF). Full article ">Figure 3
Flow chart of database searching and screening of studies for the systematic review. Full article ">

21 pages, 1746 KiB

Open AccessEditor’s ChoiceReview

Ultramicronized Palmitoylethanolamide (um-PEA): A New Possible Adjuvant Treatment in COVID-19 patients

by Annalisa Noce, Maria Albanese, Giulia Marrone, Manuela Di Lauro, Anna Pietroboni Zaitseva, Daniela Palazzetti, Cristina Guerriero, Agostino Paolino, Giuseppa Pizzenti, Francesca Di Daniele, Annalisa Romani, Cartesio D’Agostini, Andrea Magrini, Nicola Biagio Mercuri and Nicola Di Daniele

Pharmaceuticals 2021, 14(4), 336; https://doi.org/10.3390/ph14040336 - 6 Apr 2021

Cited by 24 | Viewed by 9816

Abstract

The Coronavirus Disease-19 (COVID-19) pandemic has caused more than 100,000,000 cases of coronavirus infection in the world in just a year, of which there were 2 million deaths. Its clinical picture is characterized by pulmonary involvement that culminates, in the most severe cases, [...] Read more.

The Coronavirus Disease-19 (COVID-19) pandemic has caused more than 100,000,000 cases of coronavirus infection in the world in just a year, of which there were 2 million deaths. Its clinical picture is characterized by pulmonary involvement that culminates, in the most severe cases, in acute respiratory distress syndrome (ARDS). However, COVID-19 affects other organs and systems, including cardiovascular, urinary, gastrointestinal, and nervous systems. Currently, unique-drug therapy is not supported by international guidelines. In this context, it is important to resort to adjuvant therapies in combination with traditional pharmacological treatments. Among natural bioactive compounds, palmitoylethanolamide (PEA) seems to have potentially beneficial effects. In fact, the Food and Drug Administration (FDA) authorized an ongoing clinical trial with ultramicronized (um)-PEA as an add-on therapy in the treatment of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) infection. In support of this hypothesis, in vitro and in vivo studies have highlighted the immunomodulatory, anti-inflammatory, neuroprotective and pain-relieving effects of PEA, especially in its um form. The purpose of this review is to highlight the potential use of um-PEA as an adjuvant treatment in SARS-CoV-2 infection. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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11 pages, 2339 KiB

Open AccessReview

Rapamycin: Drug Repurposing in SARS-CoV-2 Infection

by Jiri Patocka, Kamil Kuca, Patrik Oleksak, Eugenie Nepovimova, Martin Valis, Michal Novotny and Blanka Klimova

Pharmaceuticals 2021, 14(3), 217; https://doi.org/10.3390/ph14030217 - 5 Mar 2021

Cited by 35 | Viewed by 4552

Abstract

Since December 2019, SARS-CoV-2 (COVID-19) has been a worldwide pandemic with enormous consequences for human health and the world economy. Remdesivir is the only drug in the world that has been approved for the treating of COVID-19. This drug, as well as vaccination, [...] Read more.

Since December 2019, SARS-CoV-2 (COVID-19) has been a worldwide pandemic with enormous consequences for human health and the world economy. Remdesivir is the only drug in the world that has been approved for the treating of COVID-19. This drug, as well as vaccination, still has uncertain effectiveness. Drug repurposing could be a promising strategy how to find an appropriate molecule: rapamycin could be one of them. The authors performed a systematic literature review of available studies on the research describing rapamycin in association with COVID-19 infection. Only peer-reviewed English-written articles from the world’s acknowledged databases Web of Science, PubMed, Springer and Scopus were involved. Five articles were eventually included in the final analysis. The findings indicate that rapamycin seems to be a suitable candidate for drug repurposing. In addition, it may represent a better candidate for COVID-19 therapy than commonly tested antivirals. It is also likely that its efficiency will not be reduced by the high rate of viral RNA mutation. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Selection workflow. Full article ">Figure 2
The structure of rapamycin. Full article ">Figure 3
(A): Structure of SARS-CoV-2 with structural proteins. (B): Detail of SARS-CoV-2 spike protein interaction with ACE2 receptor of the host cell. (C): Top: Genome structure of the SARS-CoV-2 involves two genes ORF1a (yellow) and ORF1b (orange) encoding 16 non-structural proteins. Genes encoding four main structural proteins—spike (S), membrane (M), envelope (E) and nucleocapsid (N) are colored green. Bottom left: Major structural domains of SARS-CoV-2 S protein—signal peptide (SP), N-terminal domain (NTD), receptor-binding domain (RBD), fusion peptide (FP), heptad repeat regions 1 (HR1) and 2 (HR2), transmembrane (TM), cytoplasmic tail region (CP). Bottom right: Major structural domains of SARS-CoV-2 N protein—NTD, serine-arginine (SR)-rich domain, C-terminal domain (CTD). Rapamycin´s target domains are highlighted. (D): Simplified lifecycle of SARS-CoV-2: (1) TMPRS2 receptor activates spike glycoprotein of SARS-CoV-2 toward binding to ACE2 receptor on surface of the host cell. (2) Endocytosis of SARS-CoV-2 into the host cell. (3) Release of the viral RNA. (4) Viral RNA is translated into RNA-dependent RNA polymerase (RdRp). (5) RNA-dependent RNA polymerase synthetizes (−)-sense genomic RNA, a template for synthesis of (+)-sense genomic or subgenomic RNA. (6A) Translation of the viral structural protein N is performed in cytoplasm, while proteins S, M and E are translated at endoplasmic reticulum. (6B) Viral RNA and N protein form RNA-N complex. (7) Endoplasmic reticulum-Golgi intermediate complex (ERGIC) with inserted S, M and E proteins and RNA-N complex interact to assembly of the virion. (8) Mature virion is released from the host cell via exocytosis. Assumed rapamycin-mediated inhibitions are highlighted. Figure was created with <a href="http://BioRender.com" target="_blank">BioRender.com</a>. Full article ">Figure 4
The mTOR signaling pathway and its important regulatory functions are shown. Activated mTORC1 results in increased protein synthesis via downstream effectors S6K1 and 4EBP1. AKT-mediated activation of NF-κB increases gene expression followed by cytokine and chemokine production. Rapamycin acts as the inhibitor of mTORC1 (acute inhibition) and mTORC2 (chronic inhibition) that is crucial for affecting of the downstream pathway. Abbreviations: 4EBP1, eukaryotic translation initiation factor 4E-binding protein 1; AKT, protein kinase B; AMPK, 5’-adenosine monophosphate-activated protein kinase; CaMKKβ, Ca2+/calmodulin-dependent protein kinase kinase-β; DEPTOR, DEP domain-containing mTOR-interacting protein; FKBP12, FK506-binding protein of 12 kDa; GF, growth factors; IRS1, insulin receptor substrate 1; LKB1, liver kinase B1; NF-κB, nuclear factor kappa-light-chain-enhancer of activated B cells; mLST8, mammalian lethal with SEC13 protein 8; mSIN1, mammalian SAPK interacting protein 1; PDK1, 3-phosphoinositide-dependent protein kinase 1; PI3K, phosphatidylinositol 3-kinase; PIP2, phosphatidylinositol (4,5)-bisphosphate; PIP3, phosphatidylinositol (3,4,5)-trisphosphate; PKC-α, protein kinase C alpha; PRAS40, proline-rich AKT substrate of 40 kDa; PROTOR, protein observed with rictor; RAPTOR, regulatory associated protein of mTOR; Rheb, ras homolog enriched in brain; RICTOR, rapamycin-insensitive companion of mTOR; RTK, receptor tyrosine kinase; S6K1, ribosomal protein S6 kinase 1; SGK1, serum- and glucocorticoid-induced kinase 1; SREBP, sterol regulatory element binding protein 1; TAK1, TGF-β activated kinase 1; TEL2, telomere maintenance 2; TFEB, transcription factor EB; TSC1 and 2, tuberous sclerosis complex 1 and 2; TTI1, TEL2-interacting protein 1; ULK1, Unc-51 like autophagy activating kinase. Figure was created with <a href="http://BioRender.com" target="_blank">BioRender.com</a>. Full article ">

14 pages, 1933 KiB

Open AccessReview

COVID-19—The Potential Beneficial Therapeutic Effects of Spironolactone during SARS-CoV-2 Infection

by Katarzyna Kotfis, Kacper Lechowicz, Sylwester Drożdżal, Paulina Niedźwiedzka-Rystwej, Tomasz K. Wojdacz, Ewelina Grywalska, Jowita Biernawska, Magda Wiśniewska and Miłosz Parczewski

Pharmaceuticals 2021, 14(1), 71; https://doi.org/10.3390/ph14010071 - 17 Jan 2021

Cited by 31 | Viewed by 11466

Abstract

In March 2020, coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2 was declared a global pandemic by the World Health Organization (WHO). The clinical course of the disease is unpredictable but may lead to severe acute respiratory infection (SARI) and pneumonia leading to acute [...] Read more.

In March 2020, coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2 was declared a global pandemic by the World Health Organization (WHO). The clinical course of the disease is unpredictable but may lead to severe acute respiratory infection (SARI) and pneumonia leading to acute respiratory distress syndrome (ARDS). It has been shown that pulmonary fibrosis may be one of the major long-term complications of COVID-19. In animal models, the use of spironolactone was proven to be an important drug in the prevention of pulmonary fibrosis. Through its dual action as a mineralocorticoid receptor (MR) antagonist and an androgenic inhibitor, spironolactone can provide significant benefits concerning COVID-19 infection. The primary effect of spironolactone in reducing pulmonary edema may also be beneficial in COVID-19 ARDS. Spironolactone is a well-known, widely used and safe anti-hypertensive and antiandrogenic medication. It has potassium-sparing diuretic action by antagonizing mineralocorticoid receptors (MRs). Spironolactone and potassium canrenoate, exerting combined pleiotropic action, may provide a therapeutic benefit to patients with COVID-19 pneumonia through antiandrogen, MR blocking, antifibrotic and anti-hyperinflammatory action. It has been proposed that spironolactone may prevent acute lung injury in COVID-19 infection due to its pleiotropic effects with favorable renin–angiotensin–aldosterone system (RAAS) and ACE2 expression, reduction in transmembrane serine protease 2 (TMPRSS2) activity and antiandrogenic action, and therefore it may prove to act as additional protection for patients at highest risk of severe pneumonia. Future prospective clinical trials are warranted to evaluate its therapeutic potential. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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34 pages, 18804 KiB

Open AccessReview

Inhibition of SARS-CoV-2 Entry into Host Cells Using Small Molecules

by Kenana Al Adem, Aya Shanti, Cesare Stefanini and Sungmun Lee

Pharmaceuticals 2020, 13(12), 447; https://doi.org/10.3390/ph13120447 - 8 Dec 2020

Cited by 26 | Viewed by 6595

Abstract

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), a virus belonging to the Coronavirus family, is now known to cause Coronavirus Disease (Covid-19) which was first recognized in December 2019. Covid-19 leads to respiratory illnesses ranging from mild infections to pneumonia and lung failure. [...] Read more.

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), a virus belonging to the Coronavirus family, is now known to cause Coronavirus Disease (Covid-19) which was first recognized in December 2019. Covid-19 leads to respiratory illnesses ranging from mild infections to pneumonia and lung failure. Strikingly, within a few months of its first report, Covid-19 has spread worldwide at an exceptionally high speed and it has caused enormous human casualties. As yet, there is no specific treatment for Covid-19. Designing inhibitory drugs that can interfere with the viral entry process constitutes one of the main preventative therapies that could combat SARS-CoV-2 infection at an early stage. In this review, we provide a brief introduction of the main features of coronaviruses, discuss the entering mechanism of SARS-CoV-2 into human host cells and review small molecules that inhibit SARS-CoV-2 entry into host cells. Specifically, we focus on small molecules, identified by experimental validation and/or computational prediction, that target the SARS-CoV-2 spike protein, human angiotensin converting enzyme 2 (ACE2) receptor and the different host cell proteases that activate viral fusion. Given the persistent rise in Covid-19 cases to date, efforts should be directed towards validating the therapeutic effectiveness of these identified small molecule inhibitors. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Figure 1
Basic structure of SARS-CoV-2 including its genome RNA and the four main structural proteins. The schematic is not drawn to scale. Enlargement has been employed to better depict the structures. Full article ">Figure 2
Entry mechanism of Sars-CoV-2 into human host cells. SARS-CoV-2 enters host cell by attaching its surface-anchored spike protein to the cell surface receptor ACE2. Such attachment activates various host cell proteases and causes them to cleave the spike protein at a site near the S1/S2 boundary. Cleavage of the spike protein, in turn, facilitates viral fusion and subsequent insertion of the viral genome into the host cell. Sites of inhibition by small molecules, namely the site of receptor binding, the site of host protein cleavage and the site of membrane fusion, are marked at each stage of viral entry. The schematic is not drawn to scale. Enlargement has been employed to better depict the viral entry mechanism. Full article ">Figure 3
SARS-CoV-2 spike protein. (A) The 3D structure of the SARS-CoV-2 trimeric spike protein colored by its monomers. (B) Magnified visualization of the secondary structure of the SARS-CoV-2 spike RBD where Beta sheets are labelled and colored in magenta, alpha helices are colored in cyan and loops are colored in pink. The RBM which directly interacts with ACE2 is colored in gray and its beta sheets are labelled. This figure was generated using PyMOL (Version 2) from the PDB: 6VSB for (A) and PDB: 6M0J for (B). Full article ">Figure 4
SARS CoV-2 spike RBD−ACE2 binding interface. (A) The resolved 3D crystal structure of the SARS CoV-2 spike RBD in contact with human ACE2 receptor (PDB: 6LZG). The ACE2 is colored in cyan and its interacting amino acids are colored in magenta while the RBD is colored in green and its interacting amino acids are colored in red. (B) Zoom-in (or enlarged) view of the ACE2 showing the residues that are involved in hydrogen binding with the RBD. (C) Zoom-in view of the RBD showing the residues that are involved in hydrogen binding with ACE2. The labelled amino acids of ACE2 and RBD in (B,C) refer to all the interacting residues that are reported in the four resolved RBD−ACE2 structures (PDBs 6VW1, 6M0J, 6M17 and 6LZG); PDB 6LZG was used as a representative only. Refer to <a href="#pharmaceuticals-13-00447-t001" class="html-table">Table 1</a> for the detailed description of hydrogen bonds formed by the highlighted residues. This figure was generated using PyMOL (Version 2). Full article ">Figure 5
Pairwise amino acid sequence alignment of the receptor binding domain (RBD) of SARS-CoV-1 (Uniprot ID: P59594) and SARS-CoV-2 (Uniprot ID: P0DTC2) performed using Clustal Omega [<a href="#B44-pharmaceuticals-13-00447" class="html-bibr">44</a>]. The asterisk * denotes positions with a single, fully conserved residue. The colon: denotes positions with conservation of groups of strongly similar properties. The period denotes conservation of groups of weakly similar properties. The region labelled in red denotes the receptor binding motif (RBM). RBD of SARS-CoV-1 and SARS-CoV-2 share a 73% identity while their RBMs share only a 50% identity. The amino acid substitutions in the RBM of SARS-CoV-2 result in a greater number of residues interacting with the host cell surface receptor. Full article ">

11 pages, 966 KiB

Open AccessReview

Role of 2-[¹⁸F]FDG as a Radiopharmaceutical for PET/CT in Patients with COVID-19: A Systematic Review

by Salvatore Annunziata, Roberto C. Delgado Bolton, Christel-Hermann Kamani, John O. Prior, Domenico Albano, Francesco Bertagna and Giorgio Treglia

Pharmaceuticals 2020, 13(11), 377; https://doi.org/10.3390/ph13110377 - 10 Nov 2020

Cited by 24 | Viewed by 3598

Abstract

Some recent studies evaluated the role of fluorine-18 fluorodeoxyglucose (2-[¹⁸F]FDG) as a radiopharmaceutical for positron emission tomography/computed tomography (PET/CT) imaging in patients with Coronavirus Disease (COVID-19). This article aims to perform a systematic review in this setting. A comprehensive computer literature [...] Read more.

Some recent studies evaluated the role of fluorine-18 fluorodeoxyglucose (2-[¹⁸F]FDG) as a radiopharmaceutical for positron emission tomography/computed tomography (PET/CT) imaging in patients with Coronavirus Disease (COVID-19). This article aims to perform a systematic review in this setting. A comprehensive computer literature search in PubMed/MEDLINE and Cochrane library databases regarding the role of 2-[¹⁸F]FDG PET/CT in patients with COVID-19 was carried out. This combination of key words was used: (A) “PET” OR “positron emission tomography” AND (B) “COVID” OR “SARS”. Only pertinent original articles were selected; case reports and very small case series were excluded. We have selected 11 original studies of 2-[¹⁸F]FDG PET/CT in patients with COVID-19. Evidence-based data showed first preliminary applications of this diagnostic tool in this clinical setting, with particular regard to the incidental detection of interstitial pneumonia suspected for COVID-19. To date, according to evidence-based data, 2-[¹⁸F]FDG PET/CT cannot substitute or integrate high-resolution CT to diagnose suspicious COVID-19 or for disease monitoring, but it can only be useful to incidentally detect suspicious COVID-19 lesions in patients performing this imaging method for standard oncological and non-oncological indications. Published data about the possible role of 2-[¹⁸F]FDG PET/CT in patients with COVID-19 are increasing, but larger studies are warranted. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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32 pages, 2554 KiB

Open AccessReview

Emerging Therapeutic Modalities against COVID-19

by Shipra Malik, Anisha Gupta, Xiaobo Zhong, Theodore P. Rasmussen, Jose E. Manautou and Raman Bahal

Pharmaceuticals 2020, 13(8), 188; https://doi.org/10.3390/ph13080188 - 8 Aug 2020

Cited by 20 | Viewed by 11392

Abstract

The novel SARS-CoV-2 virus has quickly spread worldwide, bringing the whole world as well as the economy to a standstill. As the world is struggling to minimize the transmission of this devastating disease, several strategies are being actively deployed to develop therapeutic interventions. [...] Read more.

The novel SARS-CoV-2 virus has quickly spread worldwide, bringing the whole world as well as the economy to a standstill. As the world is struggling to minimize the transmission of this devastating disease, several strategies are being actively deployed to develop therapeutic interventions. Pharmaceutical companies and academic researchers are relentlessly working to investigate experimental, repurposed or FDA-approved drugs on a compassionate basis and novel biologics for SARS-CoV-2 prophylaxis and treatment. Presently, a tremendous surge of COVID-19 clinical trials are advancing through different stages. Among currently registered clinical efforts, ~86% are centered on testing small molecules or antibodies either alone or in combination with immunomodulators. The rest ~14% of clinical efforts are aimed at evaluating vaccines and convalescent plasma-based therapies to mitigate the disease's symptoms. This review provides a comprehensive overview of current therapeutic modalities being evaluated against SARS-CoV-2 virus in clinical trials. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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17 pages, 1608 KiB

Open AccessReview

Immune Pathogenesis of COVID-19 Intoxication: Storm or Silence?

by Mikhail Kiselevskiy, Irina Shubina, Irina Chikileva, Suria Sitdikova, Igor Samoylenko, Natalia Anisimova, Kirill Kirgizov, Amina Suleimanova, Tatyana Gorbunova and Svetlana Varfolomeeva

Pharmaceuticals 2020, 13(8), 166; https://doi.org/10.3390/ph13080166 - 26 Jul 2020

Cited by 17 | Viewed by 5957

Abstract

Dysregulation of the immune system undoubtedly plays an important and, perhaps, determining role in the COVID-19 pathogenesis. While the main treatment of the COVID-19 intoxication is focused on neutralizing the excessive inflammatory response, it is worth considering an equally significant problem of the [...] Read more.

Dysregulation of the immune system undoubtedly plays an important and, perhaps, determining role in the COVID-19 pathogenesis. While the main treatment of the COVID-19 intoxication is focused on neutralizing the excessive inflammatory response, it is worth considering an equally significant problem of the immunosuppressive conditions including immuno-paralysis, which lead to the secondary infection. Therefore, choosing a treatment strategy for the immune-mediated complications of coronavirus infection, one has to pass between Scylla and Charybdis, so that, in the fight against the “cytokine storm,” it is vital not to miss the point of the immune silence that turns into immuno-paralysis. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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29 pages, 2224 KiB

Open AccessReview

Substance Use Disorder in the COVID-19 Pandemic: A Systematic Review of Vulnerabilities and Complications

by Yufeng Wei and Rameen Shah

Pharmaceuticals 2020, 13(7), 155; https://doi.org/10.3390/ph13070155 - 18 Jul 2020

Cited by 87 | Viewed by 14889

Abstract

As the world endures the coronavirus disease 2019 (COVID-19) pandemic, the conditions of 35 million vulnerable individuals struggling with substance use disorders (SUDs) worldwide have not received sufficient attention for their special health and medical needs. Many of these individuals are complicated by [...] Read more.

As the world endures the coronavirus disease 2019 (COVID-19) pandemic, the conditions of 35 million vulnerable individuals struggling with substance use disorders (SUDs) worldwide have not received sufficient attention for their special health and medical needs. Many of these individuals are complicated by underlying health conditions, such as cardiovascular and lung diseases and undermined immune systems. During the pandemic, access to the healthcare systems and support groups is greatly diminished. Current research on COVID-19 has not addressed the unique challenges facing individuals with SUDs, including the heightened vulnerability and susceptibility to the disease. In this systematic review, we will discuss the pathogenesis and pathology of COVID-19, and highlight potential risk factors and complications to these individuals. We will also provide insights and considerations for COVID-19 treatment and prevention in patients with SUDs. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Figure 1
(A) Architecture of SARS-CoV-2 genome. The ORF1ab will be translated into two overlapping polyproteins, PP1a, consisting of NSP1-11, and PP1ab, consisting of NSP1-16, with the exception of NSP11, which is part of NSP12 in PP1ab. The rest of the ORFs encode the four structural proteins, S, E, M, and N, and several accessory proteins with unknown functions. (B) Structure of SARS-CoV-2 virion. The lipid bilayer. embedded with S, E, and M proteins, capsulizes the single-stranded genomic RNA, which is stabilized by the N protein. The S protein is responsible for the recognition of host cell ACE2 receptor to gain cell entry. Full article ">Figure 2
Schematic illustration of vascular endothelial junctional architecture. SARS-CoV-2 infects vascular endothelial cells through the surface-expressed ACE2 receptor. The internalization of the virus can cause endothelial cell death, reactive oxidative species (ROS), and the release of various proinflammatory cytokines. Excessive inflammation, and potentially cytokine storm, induces the loosening of the tight junction complex and cytoskeletal remodeling, leading to vascular leakage and coagulation. Various substances of abuse exert similar effects at the brain endothelial junctions, disrupting the BBB and allowing viral infection in the CNS. Full article ">Figure 3
Bidirectional communication between the brain and the immune system. The HPA axis: upon activation (cytokines, pathogens, etc.), the hypothalamus in the brain produces CRH and AVP, activating anterior pituitary, which secretes ACTH. ACTH circulates with general blood stream to reach adrenal gland, which synthesizes the anti-inflammatory molecule, glucocorticoids. Glucocorticoids suppress the immune system and the expression of proinflammatory cytokines, which concludes the negative feedback and turns off the HPA axis. Glucocorticoids suppress the activities of various immune cells, including macrophages, dendritic cells, and T cells, which are responsible for cytokine release. The immunosuppression also involves inhibition of NK cells, B cells, and T cells for reduced cytotoxicity, antibody production, and T cell-mediated immune responses. Substances of abuse alter the HPA axis. Excessive production of glucocorticoids suppresses immune responses to viral infection, leading to high incidences of infection and severe infection in COVID-19. Arrows indicate stimulation; blunted arrows indicate inhibition. Full article ">Figure 4
Pathological effects of substances of abuse on various tissues and systems and their implied complications in COVID-19. (A) Respiratory system; (B) Cardiovascular system; (C) Vascular endothelium; (D) HPA axis stimulation and immunosuppression; (E) Proinflammation and neuroinflammation. Full article ">

29 pages, 1186 KiB

Open AccessReview

The Rationale for Potential Pharmacotherapy of COVID-19

by Maha Saber-Ayad, Mohamed A. Saleh and Eman Abu-Gharbieh

Pharmaceuticals 2020, 13(5), 96; https://doi.org/10.3390/ph13050096 - 14 May 2020

Cited by 40 | Viewed by 9255

Abstract

On 11 March 2020, the coronavirus disease (COVID-19) was defined by the World Health Organization as a pandemic. Severe acute respiratory syndrome-2 (SARS-CoV-2) is the newly evolving human coronavirus infection that causes COVID-19, and it first appeared in Wuhan, China in December 2019 [...] Read more.

On 11 March 2020, the coronavirus disease (COVID-19) was defined by the World Health Organization as a pandemic. Severe acute respiratory syndrome-2 (SARS-CoV-2) is the newly evolving human coronavirus infection that causes COVID-19, and it first appeared in Wuhan, China in December 2019 and spread rapidly all over the world. COVID-19 is being increasingly investigated through virology, epidemiology, and clinical management strategies. There is currently no established consensus on the standard of care in the pharmacological treatment of COVID-19 patients. However, certain medications suggested for other diseases have been shown to be potentially effective for treating this infection, though there has yet to be clear evidence. Therapies include new agents that are currently tested in several clinical trials, in addition to other medications that have been repurposed as antiviral and immune-modulating therapies. Previous high-morbidity human coronavirus epidemics such as the 2003 SARS-CoV and the 2012 Middle East respiratory syndrome coronavirus (MERS-CoV) prompted the identification of compounds that could theoretically be active against the emerging coronavirus SARS-CoV-2. Moreover, advances in molecular biology techniques and computational analysis have allowed for the better recognition of the virus structure and the quicker screening of chemical libraries to suggest potential therapies. This review aims to summarize rationalized pharmacotherapy considerations in COVID-19 patients in order to serve as a tool for health care professionals at the forefront of clinical care during this pandemic. All the reviewed therapies require either additional drug development or randomized large-scale clinical trials to be justified for clinical use. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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1 pages, 165 KiB

Open AccessCorrection

Correction: Awad et al. Repurposing Potential of the Antiparasitic Agent Ivermectin for the Treatment and/or Prophylaxis of COVID-19. Pharmaceuticals 2022, 15, 1068

by Hoda Awad, Basmala Hassan, Sara Dweek, Yasmeen Aboelata, Mutasem Rawas-Qalaji and Iman Saad Ahmed

Pharmaceuticals 2024, 17(8), 1100; https://doi.org/10.3390/ph17081100 - 22 Aug 2024

Viewed by 485

Abstract

The following paragraph, “Coinciding but insignificant findings with regards to the time to negative PCR were established by Pott-Junior et al [...] Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

1 pages, 174 KiB

Open AccessCorrection

Correction: Palmeira et al. Preliminary Virtual Screening Studies to Identify GRP78 Inhibitors Which May Interfere with SARS-CoV-2 Infection. Pharmaceuticals 2020, 13, 132

by Andreia Palmeira, Emília Sousa, Aylin Köseler, Ramazan Sabirli, Tarık Gören, İbrahim Türkçüer, Özgür Kurt, Madalena M. Pinto and M. Helena Vasconcelos

Pharmaceuticals 2024, 17(4), 411; https://doi.org/10.3390/ph17040411 - 25 Mar 2024

Viewed by 890

Abstract

There was an error in the original publication [...] Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

31 pages, 558 KiB

Open AccessSystematic Review

Biologics in COVID-19 So Far: Systematic Review

by Milton Arias, Henry Oliveros, Sharon Lechtig and Rosa-Helena Bustos

Pharmaceuticals 2022, 15(7), 783; https://doi.org/10.3390/ph15070783 - 23 Jun 2022

Cited by 8 | Viewed by 2658

Abstract

This systematic review aimed to reevaluate the available evidence of the use of biologics as treatment candidates for the treatment of severe and advanced COVID-19 disease; what are the rationale for their use, which are the most studied, and what kind of efficacy [...] Read more.

This systematic review aimed to reevaluate the available evidence of the use of biologics as treatment candidates for the treatment of severe and advanced COVID-19 disease; what are the rationale for their use, which are the most studied, and what kind of efficacy measures are described? A search through Cochrane, Embase, Pubmed, Medline, medrxiv.org, and Google scholar was performed on the use of biologic interventions in COVID-19/SARS-CoV-2 infection, viral pneumonia, and sepsis, until 11 January 2022. Throughout the research, we identified 4821 records, of which 90 were selected for qualitative analysis. Amongst the results, we identified five popular targets of use: IL6 and IL1 inhibitors, interferons, mesenchymal stem cells treatment, and anti-spike antibodies. None of them offered conclusive evidence of their efficacy with consistency and statistical significance except for some studies with anti-spike antibodies; however, Il6 and IL1 inhibitors as well as interferons show encouraging data in terms of increased survival and favorable clinical course that require further studies with better methodology standardization. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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1 pages, 163 KiB

Open AccessCorrection

Correction: Zanella et al. Tenofovir, Another Inexpensive, Well Known and Widely Available Old Drug Repurposed for SARS-COV-2 Infection. Pharmaceuticals 2021, 14, 454

by Isabella Zanella, Daniela Zizioli, Francesco Castelli and Eugenia Quiros-Roldan

Pharmaceuticals 2021, 14(8), 827; https://doi.org/10.3390/ph14080827 - 23 Aug 2021

Cited by 3 | Viewed by 2351

Abstract

Text Correction [...] Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

15 pages, 370 KiB

Open AccessPerspective

Update on Functional Inhibitors of Acid Sphingomyelinase (FIASMAs) in SARS-CoV-2 Infection

by Gwenolé Loas and Pascal Le Corre

Pharmaceuticals 2021, 14(7), 691; https://doi.org/10.3390/ph14070691 - 18 Jul 2021

Cited by 12 | Viewed by 3918

Abstract

The SARS-CoV-2 outbreak is characterized by the need of the search for curative drugs for treatment. In this paper, we present an update of knowledge about the interest of the functional inhibitors of acid sphingomyelinase (FIASMAs) in SARS-CoV-2 infection. Forty-nine FIASMAs have been [...] Read more.

The SARS-CoV-2 outbreak is characterized by the need of the search for curative drugs for treatment. In this paper, we present an update of knowledge about the interest of the functional inhibitors of acid sphingomyelinase (FIASMAs) in SARS-CoV-2 infection. Forty-nine FIASMAs have been suggested in the treatment of SARS-CoV-2 infection using in silico, in vitro or in vivo studies. Further studies using large-sized, randomized and double-blinded controlled clinical trials are needed to evaluate FIASMAs in SARS-CoV-2 infection as off-label therapy. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

21 pages, 2194 KiB

Open AccessOpinion

COVID-19: Failure of the DisCoVeRy Clinical Trial, and Now–New Hopes?

by Jean Jacques Vanden Eynde

Pharmaceuticals 2021, 14(7), 664; https://doi.org/10.3390/ph14070664 - 11 Jul 2021

Cited by 4 | Viewed by 4860

Abstract

The DisCoVeRy clinical trial aimed at the evaluation of four treatments for patients suffering from severe to critical COVID-19: Hydroxychloroquine, eventually associated with azithromycin; the combination lopinavir/ritonavir; the combination with the addition of interferon β-1a; remdesivir. The trial was discontinued due to the [...] Read more.

The DisCoVeRy clinical trial aimed at the evaluation of four treatments for patients suffering from severe to critical COVID-19: Hydroxychloroquine, eventually associated with azithromycin; the combination lopinavir/ritonavir; the combination with the addition of interferon β-1a; remdesivir. The trial was discontinued due to the lack of positive results. Meanwhile, many other potential options have been considered either to target the virus itself, the interactions with the host cells, or the cytokine storm frequently observed during the infection. Several of those options are briefly reviewed. They include vaccines, small molecules, antibodies, and stem cells. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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15 pages, 1003 KiB

Open AccessPerspective

Calming the (Cytokine) Storm: Dimethyl Fumarate as a Therapeutic Candidate for COVID-19

by Cara A. Timpani and Emma Rybalka

Pharmaceuticals 2021, 14(1), 15; https://doi.org/10.3390/ph14010015 - 26 Dec 2020

Cited by 28 | Viewed by 5809

Abstract

COVID-19 has rapidly spread worldwide and incidences of hospitalisation from respiratory distress are significant. While a vaccine is in the pipeline, there is urgency for therapeutic options to address the immune dysregulation, hyperinflammation and oxidative stress that can lead to death. Given the [...] Read more.

COVID-19 has rapidly spread worldwide and incidences of hospitalisation from respiratory distress are significant. While a vaccine is in the pipeline, there is urgency for therapeutic options to address the immune dysregulation, hyperinflammation and oxidative stress that can lead to death. Given the shared pathogenesis of severe cases of COVID-19 with aspects of multiple sclerosis and psoriasis, we propose dimethyl fumarate as a viable treatment option. Currently approved for multiple sclerosis and psoriasis, dimethyl fumarate is an immunomodulatory, anti-inflammatory and anti-oxidative drug that could be rapidly implemented into the clinic to calm the cytokine storm which drives severe COVID-19. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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Full article ">Figure 1
Simplified pathological pathway to lung (and systemic) injury in severe cases of COVID-19. Infection with the SARS-CoV-2 virus can lead to a dysregulated immune response in which pro-inflammatory cells dominate the immune cell population. These pro-inflammatory cells intensify cytokine production and release resulting in hyperinflammation. This hyperinflammatory state promotes lung (and systemic) pathology, which correlates with poorer prognosis. It is well documented that dimethyl fumarate (DMF) can modulate immune cell populations to shift the ratio of anti-inflammatory to pro-inflammatory cytokine production and release, which in turn reduces hyperinflammation and subsequent tissue injury. Full article ">Figure 2
Simplified schematic of pathways activated by dimethyl fumarate (DMF). DMF is known to mediate its anti-inflammatory, anti-oxidative and immunomodulatory effects primarily through three molecular pathways: (1) Activation of nuclear factor erythroid 2-related factor 2 (Nrf2) which binds to the antioxidant response element (ARE) in the nucleus to stimulate transcription of Phase II enzymes including superoxide dismutase (SOD1), NAD(P)H quinone oxidoreductase-1 (NQO1) and heme oxygenase-1 (HO-1). Together, SOD1, NQO1 and HO-1 mediate cytoprotective, anti-oxidative and anti-inflammatory effects. (2) DMF can directly inhibit nuclear factor kappa B (NF-κB), which prevents the translocation of NF-κB into the nucleus, binding to the κB site, release of pro-inflammatory cytokines and subsequent inflammation and damage. DMF can also indirectly inhibit NF-κB through HO-1 expression and immune cell modulation through hydroxycarboxylic acid receptor 2 (HCAR2) activation. (3) DMF’s immunomodulatory effects are predominantly mediated through HCAR2 activation which modulates immune cell populations (pro-inflammatory to anti-inflammatory shift) and inhibits NF-κB. Adapted from [<a href="#B7-pharmaceuticals-14-00015" class="html-bibr">7</a>]. Created with BioRender.com. Full article ">

17 pages, 371 KiB

Open AccessCommentary

Emetine Is Not Ipecac: Considerations for Its Use as Treatment for SARS-CoV2

by Martin D. Bleasel and Gregory M. Peterson

Pharmaceuticals 2020, 13(12), 428; https://doi.org/10.3390/ph13120428 - 27 Nov 2020

Cited by 14 | Viewed by 5525

Abstract

Emetine is a potent antiviral that acts on many viruses in the low-nM range, with several studies in animals and humans demonstrating antiviral activity. Historically, emetine was used to treat patients with Spanish influenza, in the last stages of the pandemic in the [...] Read more.

Emetine is a potent antiviral that acts on many viruses in the low-nM range, with several studies in animals and humans demonstrating antiviral activity. Historically, emetine was used to treat patients with Spanish influenza, in the last stages of the pandemic in the early 1900s. Some of these patients were “black” with cyanosis. Emetine rapidly reversed the cyanosis and other symptoms of this disease in 12–24 h. However, emetine also has been shown to have anti-inflammatory properties and it appears it is these anti-inflammatory properties that were responsible for the effects seen in patients with Spanish influenza. Emetine, in the past, has also been used in 10s to 100s of millions of people at a dose of ~60 mg daily to treat amoebiasis. Based on viral inhibition data we can calculate a likely SARS-CoV2 antiviral dose of ~1/10th the amoebiasis dose, which should dramatically reduce the risk of any side effects. While there are no anti-inflammatory dose response data available, based on the potential mode of action, the anti-inflammatory actions may also occur at low doses. This paper also examines the toxicity of emetine seen in clinical practice and that seen in the laboratory, and discusses the methods of administration aimed at reducing side effects if higher doses were found to be necessary. While emetine is a “pure drug” as it is extracted from ipecac, some of the differences between emetine and ipecac are also discussed. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

8 pages, 980 KiB

Open AccessOpinion

Can Nuclear Imaging of Activated Macrophages with Folic Acid-Based Radiotracers Serve as a Prognostic Means to Identify COVID-19 Patients at Risk?

by Cristina Müller, Roger Schibli and Britta Maurer

Pharmaceuticals 2020, 13(9), 238; https://doi.org/10.3390/ph13090238 - 9 Sep 2020

Cited by 9 | Viewed by 3910

Abstract

Herein, we discuss the potential role of folic acid-based radiopharmaceuticals for macrophage imaging to support clinical decision-making in patients with COVID-19. Activated macrophages play an important role during coronavirus infections. Exuberant host responses, i.e., a cytokine storm with increase of macrophage-related cytokines, such [...] Read more.

Herein, we discuss the potential role of folic acid-based radiopharmaceuticals for macrophage imaging to support clinical decision-making in patients with COVID-19. Activated macrophages play an important role during coronavirus infections. Exuberant host responses, i.e., a cytokine storm with increase of macrophage-related cytokines, such as TNFα, IL-1β, and IL-6 can lead to life-threatening complications, such as acute respiratory distress syndrome (ARDS), which develops in approximately 20% of the patients. Diverse immune modulating therapies are currently being tested in clinical trials. In a preclinical proof-of-concept study in experimental interstitial lung disease, we showed the potential of ¹⁸F-AzaFol, an ¹⁸F-labeled folic acid-based radiotracer, as a specific novel imaging tool for the visualization and monitoring of macrophage-driven lung diseases. ¹⁸F-AzaFol binds to the folate receptor-beta (FRβ) that is expressed on activated macrophages involved in inflammatory conditions. In a recent multicenter cancer trial, ¹⁸F-AzaFol was successfully and safely applied (NCT03242993). It is supposed that the visualization of activated macrophage-related disease processes by folate radiotracer-based nuclear imaging can support clinical decision-making by identifying COVID-19 patients at risk of a severe disease progression with a potentially lethal outcome. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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14 pages, 601 KiB

Open AccessOpinion

Cancer Patients Have a Higher Risk Regarding COVID-19–and Vice Versa?

by Franz Geisslinger, Angelika M. Vollmar and Karin Bartel

Pharmaceuticals 2020, 13(7), 143; https://doi.org/10.3390/ph13070143 - 6 Jul 2020

Cited by 17 | Viewed by 5360

Abstract

The world is currently suffering from a pandemic which has claimed the lives of over 230,000 people to date. The responsible virus is called severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and causes the coronavirus disease 2019 (COVID-19), which is mainly characterized by [...] Read more.

The world is currently suffering from a pandemic which has claimed the lives of over 230,000 people to date. The responsible virus is called severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and causes the coronavirus disease 2019 (COVID-19), which is mainly characterized by fever, cough and shortness of breath. In severe cases, the disease can lead to respiratory distress syndrome and septic shock, which are mostly fatal for the patient. The severity of disease progression was hypothesized to be related to an overshooting immune response and was correlated with age and comorbidities, including cancer. A lot of research has lately been focused on the pathogenesis and acute consequences of COVID-19. However, the possibility of long-term consequences caused by viral infections which has been shown for other viruses are not to be neglected. In this regard, this opinion discusses the interplay of SARS-CoV-2 infection and cancer with special focus on the inflammatory immune response and tissue damage caused by infection. We summarize the available literature on COVID-19 suggesting an increased risk for severe disease progression in cancer patients, and we discuss the possibility that SARS-CoV-2 could contribute to cancer development. We offer lines of thought to provide ideas for urgently needed studies on the potential long-term effects of SARS-CoV-2 infection. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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9 pages, 1321 KiB

Open AccessOpinion

COVID-19: An Update about the Discovery Clinical Trial

by Jean Jacques Vanden Eynde

Pharmaceuticals 2020, 13(5), 98; https://doi.org/10.3390/ph13050098 - 14 May 2020

Cited by 12 | Viewed by 5708

Abstract

Finding efficacious and safe treatments for COVID-19 emerges as a crucial need in order to control the spread of the pandemic. Whereas plasma therapy attracts much interest, the European project Discovery focuses on the potentialities of small molecules like remdesivir, the combination of [...] Read more.

Finding efficacious and safe treatments for COVID-19 emerges as a crucial need in order to control the spread of the pandemic. Whereas plasma therapy attracts much interest, the European project Discovery focuses on the potentialities of small molecules like remdesivir, the combination of lopinavir/ritonavir, hydroxychloroquine, and chloroquine. Results recently published on the clinical evaluation of those drugs are compiled in this brief report, although complete data are still impatiently awaited. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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8 pages, 654 KiB

Open AccessOpinion

COVID-19: A Brief Overview of the Discovery Clinical Trial

by Jean Jacques Vanden Eynde

Pharmaceuticals 2020, 13(4), 65; https://doi.org/10.3390/ph13040065 - 10 Apr 2020

Cited by 30 | Viewed by 10912

Abstract

The outbreak of COVID-19 is leading to a tremendous search for curative treatments. The urgency of the situation favors a repurposing of active drugs but not only antivirals. This short communication focuses on four treatments recommended by WHO and included in the first [...] Read more.

The outbreak of COVID-19 is leading to a tremendous search for curative treatments. The urgency of the situation favors a repurposing of active drugs but not only antivirals. This short communication focuses on four treatments recommended by WHO and included in the first clinical trial of the European Discovery project. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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9 pages, 259 KiB

Open AccessCommentary

Emetine, Ipecac, Ipecac Alkaloids and Analogues as Potential Antiviral Agents for Coronaviruses

by Martin D. Bleasel and Gregory M. Peterson

Pharmaceuticals 2020, 13(3), 51; https://doi.org/10.3390/ph13030051 - 21 Mar 2020

Cited by 62 | Viewed by 11716

Abstract

The COVID-19 coronavirus is currently spreading around the globe with limited treatment options available. This article presents the rationale for potentially using old drugs (emetine, other ipecac alkaloids or analogues) that have been used to treat amoebiasis in the treatment of COVID-19. Emetine [...] Read more.

The COVID-19 coronavirus is currently spreading around the globe with limited treatment options available. This article presents the rationale for potentially using old drugs (emetine, other ipecac alkaloids or analogues) that have been used to treat amoebiasis in the treatment of COVID-19. Emetine had amongst the lowest reported half-maximal effective concentration (EC₅₀) from over 290 agents screened for the Middle East respiratory syndrome (MERS) and severe acute respiratory syndrome (SARS) coronaviruses. While EC₅₀ concentrations of emetine are achievable in the blood, studies show that concentrations of emetine can be almost 300 times higher in the lungs. Furthermore, based on the relative EC₅₀s of emetine towards the coronaviruses compared with Entamoeba histolytica, emetine could be much more effective as an anti-coronavirus agent than it is against amoebiasis. This paper also discusses the known side effects of emetine and related compounds, how those side effects can be managed, and the optimal method of administration for the potential treatment of COVID-19. Given the serious and immediate threat that the COVID-19 coronavirus poses, our long history with emetine and the likely ability of emetine to reach therapeutic concentrations within the lungs, ipecac, emetine, and other analogues should be considered as potential treatment options, especially if in vitro studies confirm viral sensitivity. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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