Bromelain Inhibits SARS-CoV-2 Infection in VeroE6 Cells

Sagar, Satish; Rathinavel, Ashok Kumar; Lutz, William E.; Struble, Lucas R.; Khurana, Surender; Schnaubelt, Andy; Mishra, Nitish K.; Guda, Chittibabu; Broadhurst, M. Jana; Reid, St Patrick; Bayles, Kenneth W.; Borgstahl, Gloria E. O.; Radhakrishnan, Prakash

doi:10.1101/2020.09.16.297366

Cited by 18 publications

(18 citation statements)

References 32 publications

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“…It is a glycoprotein with an oligosaccharide in its molecular structure. It was shown recently that bromelain inhibits SARS-CoV-2 infection in VeroE6 Cells [66]. It seems that our findings are consistent with these results.…”

Section: Nhcsupporting

confidence: 94%

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Water-SARS-CoV-2 Interaction-Based Mechanism Inhibiting Virus Attachment to Host Cells

Shalatonin

2020

Preprint

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Many studies showed that the enveloped viruses, including coronaviruses, HIV-1, and influenza inactivate significantly faster in water than the non-enveloped viruses. It looks that there is a certain mechanism controlling this phenomenon. However, the epidemic spread of SARS-CoV-2 indicates that this virus – water interaction mechanism is not effective enough to fully inhibit coronavirus reproduction. We hypothesized that a spatially extended layer of the ordered water molecules, formed around CoV due to the spike’s glycans – surrounding water interaction acts as a buffer inhibiting CoV motion and its attachment to the host cell receptor. There is experimental evidence that water molecules while interacting with glucans experience the long-range ordering and repulsive forces. Our findings revealed new features that can promote its interaction. It was shown that the glycans and water molecules have the same far ultraviolet (UV) absorbance peak at ~185 nm. This peak is a manifestation of the still little-known physical properties possessed by hydroxyl (OH) groups, including those contained in glycans and water molecules. Many studies show that the carbohydrate hydroxyl groups are a key element in the long-range antifreeze glycoproteins activity which is closely correlated with our issue. To further increase ice inhibition, a sugar-based (usually trehalose) water solution, further slowing down the water dynamics is commonly used. Our experiments with sugar-based compounds dissolved in water showed that in such solutions the UV absorbance at ~185 nm (activity of the OH groups) can be essentially increased with respect to the bulk water. The spike’s glycans – water long-range interaction, activated due to the dissolved sugar-based compound, creates the glass-like stabilizing hydration layer (like in case of the trehalose) effectively inhibiting the virus – host cell binding. It was shown that the chemical structures of the known compounds with proven inhibition of SARS-CoV-2 entry into host cells agree with our findings. The described approach can be effective against human immunodeficiency viruses, influenza viruses, and possibly other enveloped viruses. <br>

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Section: Nhcsupporting

confidence: 94%

“…To check how our findings correlate with the known compounds for COVID-19 treatment, several of them have been analyzed briefly. We directed attention to Chloroquine, Emodin, β-D-N4-Hydroxycytidine (NHC), bromelain, luteolin [61][62][63][64][65][66], and tetra-O-galloyl-β-d-glucose (TGG) [59].…”

Section: Chemical Structures Of the Antiviral Compoundsmentioning

confidence: 99%

Water-SARS-CoV-2 Interaction-Based Mechanism Inhibiting Virus Attachment to Host Cells

Shalatonin

2020

Preprint

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“…In our study, we showed for the first time a biochemical link between LL-37 and the SARS-CoV-2 S protein and provide a rationale for the use of vitamin D in infection prophylaxis, and for therapeutic administration in the treatment of COVID-19 patients. Peptide based therapeutics, especially antimicrobial peptides, are excellent drug candidates as they have little adverse effects and show anti-coronavirus activity [22][23][24] .…”

Section: Discussionmentioning

confidence: 99%

LL-37 fights SARS-CoV-2: The Vitamin D-Inducible Peptide LL-37 Inhibits Binding of SARS-CoV-2 Spike Protein to its Cellular Receptor Angiotensin Converting Enzyme 2In Vitro

Roth

Luetke

Meinberger

et al. 2020

Preprint

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Objective: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the pathogen accountable for the coronavirus disease 2019 (COVID-19) pandemic. Viral entry via binding of the receptor binding domain (RBD) located within the S1 subunit of the SARS-CoV-2 Spike (S) protein to its target receptor angiotensin converting enzyme (ACE) 2 is a key step in cell infection. The efficient transition of the virus is linked to a unique protein called open reading frame (ORF) 8. As SARS-CoV-2 infections can develop into life threatening lower respiratory syndromes, effective therapy options are urgently needed. Several publications propose vitamin D treatment, although its mode of action against COVID-19 is not fully elucidated. It is speculated that vitamin D's beneficial effects are mediated by up regulating LL-37, a well known antimicrobial peptide with antiviral effects. Methods: Recombinantly expressed SARS-CoV-2 S protein, the extended S1 subunit (S1e), the S2 subunit (S2), the receptor binding domain (RBD), and ORF8 were used for surface plasmon resonance (SPR) studies to investigate LL-37's ability to bind to SARS-CoV-2 proteins and to localize its binding site within the S protein. Binding competition studies were conducted to confirm an inhibitory action of LL-37 on the attachment of SARS-CoV-2 S protein to its entry receptor ACE2. Results: We could show that LL-37 binds to SARS-CoV-2 S protein (LL-37/S-Strep KD = 407 nM, LL-37/S-His KD = 414 nM) with the same affinity, as SARS-CoV-2 binds to hACE2 (hACE2/S-Strep KD = 374 nM, hACE2/S-His KD = 368 nM). The binding is not restricted to the RBD of the S protein, but rather distributed along the entire length of the protein. Interaction between LL-37 and ORF8 was detected with a KD of 294 nM. Further, inhibition of the binding of S-Strep (IC50 = 735 nM), S1e (IC50 = 168 nM), and RBD (IC50 = 126 nM) to hACE2 by LL-37 was demonstrated. Conclusions: We have revealed a biochemical link between vitamin D, LL-37, and COVID-19 severity. SPR analysis demonstrated that LL-37 binds to SARS-CoV-2 S protein and inhibits binding to its receptor hACE2, and most likely viral entry into the cell. This study supports the prophylactic use of vitamin D to induce LL-37 that protects from SARS-CoV-2 infection, and the therapeutic administration of vitamin D for the treatment of COVID-19 patients. Further, our results provide evidence that the direct use of LL-37 by inhalation and systemic application may reduce the severity of COVID-19.

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“…Apart from its nutritional value, bromelain’s active ingredients have been shown to have therapeutic properties, such as antiviral (e.g. anti-SARS-CoV-2), anti-inflammatory, anticancer, antimicrobial, and immunomodulatory function ( Dopheide and Ward, 1981 ; Chandler and Mynott, 1998 ; Rathnavelu et al, 2016 ; Sagar et al, 2020 ; Chakraborty et al, 2021 ). Bromelain has a half-life of 6–9 h ( Castell et al, 1997 ) and a plasma concentration of 2.5–4 ng/ml after an oral dose of 8.6 g per day ( Kumakura et al, 1988 ).…”

Section: Introductionmentioning

confidence: 99%

An Analysis Based on Molecular Docking and Molecular Dynamics Simulation Study of Bromelain as Anti-SARS-CoV-2 Variants

et al. 2021

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The rapid spread of a novel coronavirus known as SARS-CoV-2 has compelled the entire world to seek ways to weaken this virus, prevent its spread and also eliminate it. However, no drug has been approved to treat COVID-19. Furthermore, the receptor-binding domain (RBD) on this viral spike protein, as well as several other important parts of this virus, have recently undergone mutations, resulting in new virus variants. While no treatment is currently available, a naturally derived molecule with known antiviral properties could be used as a potential treatment. Bromelain is an enzyme found in the fruit and stem of pineapples. This substance has been shown to have a broad antiviral activity. In this article, we analyse the ability of bromelain to counteract various variants of the SARS-CoV-2 by targeting bromelain binding on the side of this viral interaction with human angiotensin-converting enzyme 2 (hACE2) using molecular docking and molecular dynamics simulation approaches. We have succeeded in making three-dimensional configurations of various RBD variants using protein modelling. Bromelain exhibited good binding affinity toward various variants of RBDs and binds right at the binding site between RBDs and hACE2. This result is also presented in the modelling between Bromelain, RBD, and hACE2. The molecular dynamics (MD) simulations study revealed significant stability of the bromelain and RBD proteins separately up to 100 ns with an RMSD value of 2 Å. Furthermore, despite increases in RMSD and changes in Rog values of complexes, which are likely due to some destabilized interactions between bromelain and RBD proteins, two proteins in each complex remained bonded, and the site where the two proteins bind remained unchanged. This finding indicated that bromelain could have an inhibitory effect on different SARS-CoV-2 variants, paving the way for a new SARS-CoV-2 inhibitor drug. However, more in vitro and in vivo research on this potential mechanism of action is required.

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Bromelain Inhibits SARS-CoV-2 Infection in VeroE6 Cells

Cited by 18 publications

References 32 publications

Water-SARS-CoV-2 Interaction-Based Mechanism Inhibiting Virus Attachment to Host Cells

Water-SARS-CoV-2 Interaction-Based Mechanism Inhibiting Virus Attachment to Host Cells

LL-37 fights SARS-CoV-2: The Vitamin D-Inducible Peptide LL-37 Inhibits Binding of SARS-CoV-2 Spike Protein to its Cellular Receptor Angiotensin Converting Enzyme 2In Vitro

An Analysis Based on Molecular Docking and Molecular Dynamics Simulation Study of Bromelain as Anti-SARS-CoV-2 Variants

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