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Potential Mechanism of Curcumin and Resveratrol against SARS-CoV-2

Wu et al., Research Square, doi:10.21203/rs.3.rs-2780614/v1

Apr 2023

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Curcumin for COVID-19

15th treatment shown to reduce risk in February 2021

^*, now with p = 0.0000000096 from 27 studies.

Lower risk for mortality, ventilation, hospitalization, progression, recovery, and viral clearance.

No treatment is 100% effective. Protocols combine treatments. ^* >10% efficacy, ≥3 studies.

4,500+ studies for 81 treatments. c19early.org

In Vitro study showing that curcumin and resveratrol inhibit SARS-CoV-2 infection through multiple mechanisms. Curcumin and resveratrol inhibit SARS-CoV-2 pseudovirus cell entry in HEK293-ACE2 cells with IC₅₀ values of 18.02 μM and 8.76 μM, respectively. Combined treatment further reduces pseudovirus entry. Both compounds also inhibit activity of the SARS-CoV-2 3CL protease with IC₅₀ values around 10-15 μM. Spike protein-induced cytokine storm is alleviated by curcumin or resveratrol through NFKB pathway inhibition in HEK293-ACE2 cells. Similarly, spike protein-mediated oxidative stress is reduced by the compounds via enhanced antioxidant system activity and ROS scavenging. Authors conclude that curcumin and resveratrol may help prevent and treat COVID-19 by inhibiting viral entry, replication, cytokine storm, and oxidative stress.

In Silico study showing potential benefits of curcumin in preventing severe COVID-19 manifestations by protecting mitochondria. Authors identified five mitochondrial dysfunction biomarkers (RECQL4, PYCR1, PIF1, POLQ, GLDC) associated with metabolic and immune dysregulation in severe COVID-19. Curcumin exhibited regulatory effects on these biomarkers and protected cells against SARS-CoV-2 spike protein-induced mitochondrial damage and oxidative stress. The study provides evidence for curcumin's ability to safeguard mitochondrial function, which may help prevent progression to severe disease.

45 preclinical studies support the efficacy of curcumin for COVID-19:

24 In Silico studies1-24

20 In Vitro studies1,5,7,11,17,21,25-38

1 In Vivo animal study5

In Silico studies predict inhibition of SARS-CoV-2 with curcumin or metabolites via binding to the spikeA,5,10,12,18,21 (and specifically the receptor binding domainB,8,11,14), M^proC,5,7,9-11,13,14,16,19,21,22,24,35, RNA-dependent RNA polymeraseD,11,20, ACE2E,12,13,15, nucleocapsidF,6,23, nsp10G,23, and helicaseH,25 proteins. In Vitro studies demonstrate inhibition of the spikeA,30 (and specifically the receptor binding domainB,38), M^proC,17,30,35,37, ACE2E,38, and TMPRSS2I,38 proteins. In Vitro studies demonstrate efficacy in Calu-3J,36, A549K,30, 293TL,1, HEK293-hACE2M,17,28, 293T/hACE2/TMPRSS2N,29, Vero E6O,7,11,21,28,30-32,34,36, and SH-SY5YP,27 cells. Curcumin is predicted to inhibit the interaction between the SARS-CoV-2 spike protein receptor binding domain and the human ACE2 receptor for the delta and omicron variants8, decreases pro-inflammatory cytokines induced by SARS-CoV-2 in peripheral blood mononuclear cells34, and alleviates SARS-CoV-2 spike protein-induced mitochondrial membrane damage and oxidative stress1.

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1. Zhang et al., Computational Discovery of Mitochondrial Dysfunction Biomarkers in Severe SARS-CoV-2 Infection: Facilitating Pytomedicine Screening, Phytomedicine, doi:10.1016/j.phymed.2024.155784.sciencedirect.com, doi.org.

2. Öztürkkan et al., In Silico investigation of the effects of curcuminoids on the spike protein of the omicron variant of SARS-CoV-2, Baku State University Journal of Chemistry and Material Sciences, 1:2, bsuj.bsu.edu.az/uploads/pdf/ec4204d62f7802de54e6092bf7860029.pdf.edu.az.

3. Yunze et al., Therapeutic effect and potential mechanism of curcumin, an active ingredient in Tongnao Decoction, on COVID-19 combined with stroke: a network pharmacology study and GEO database mining, Research Square, doi:10.21203/rs.3.rs-4329762/v1.researchsquare.com, doi.org.

4. Agamah et al., Network-based multi-omics-disease-drug associations reveal drug repurposing candidates for COVID-19 disease phases, ScienceOpen, doi:10.58647/DRUGARXIV.PR000010.v1.scienceopen.com, doi.org.

5. Boseila et al., Throat spray formulated with virucidal Pharmaceutical excipients as an effective early prophylactic or treatment strategy against pharyngitis post-exposure to SARS CoV-2, European Journal of Pharmaceutics and Biopharmaceutics, doi:10.1016/j.ejpb.2024.114279.sciencedirect.com, doi.org.

6. Hidayah et al., Bioinformatics study of curcumin, demethoxycurcumin, bisdemethoxycurcumin and cyclocurcumin compounds in Curcuma longa as an antiviral agent via nucleocapsid on SARS-CoV-2 inhibition, International Conference on Organic and Applied Chemistry, doi:10.1063/5.0197724.aip.org, doi.org.

7. Singh et al., Unlocking the potential of phytochemicals in inhibiting SARS-CoV-2 M Pro protein - An in-silico and cell-based approach, Research Square, doi:10.21203/rs.3.rs-3888947/v1.researchsquare.com, doi.org.

8. Kant et al., Structure-based drug discovery to identify SARS-CoV2 spike protein–ACE2 interaction inhibitors, Journal of Biomolecular Structure and Dynamics, doi:10.1080/07391102.2023.2300060.tandfonline.com, doi.org.

9. Naderi Beni et al., In silico studies of anti-oxidative and hot temperament-based phytochemicals as natural inhibitors of SARS-CoV-2 Mpro, PLOS ONE, doi:10.1371/journal.pone.0295014.plos.org, doi.org.

10. Moschovou et al., Exploring the Binding Effects of Natural Products and Antihypertensive Drugs on SARS-CoV-2: An In Silico Investigation of Main Protease and Spike Protein, International Journal of Molecular Sciences, doi:10.3390/ijms242115894.mdpi.com, doi.org.

11. Eleraky et al., Curcumin Transferosome-Loaded Thermosensitive Intranasal in situ Gel as Prospective Antiviral Therapy for SARS-Cov-2, International Journal of Nanomedicine, doi:10.2147/IJN.S423251.dovepress.com, doi.org.

12. Singh (B) et al., Computational studies to analyze effect of curcumin inhibition on coronavirus D614G mutated spike protein, The Seybold Report, doi:10.17605/OSF.IO/TKEXJ.ac.in, doi.org.

13. Thapa et al., In-silico Approach for Predicting the Inhibitory Effect of Home Remedies on Severe Acute Respiratory Syndrome Coronavirus-2, Makara Journal of Science, doi:10.7454/mss.v27i3.1609.ac.id, doi.org.

14. Srivastava et al., Paradigm of Well-Orchestrated Pharmacokinetic Properties of Curcuminoids Relative to Conventional Drugs for the Inactivation of SARS-CoV-2 Receptors: An In Silico Approach, Stresses, doi:10.3390/stresses3030043.mdpi.com, doi.org.

15. Alkafaas et al., A study on the effect of natural products against the transmission of B.1.1.529 Omicron, Virology Journal, doi:10.1186/s12985-023-02160-6.biomedcentral.com, doi.org.

16. Winih Kinasih et al., Analisis in silico interaksi senyawa kurkuminoid terhadap enzim main protease 6LU7 dari SARS-CoV-2, Duta Pharma Journal, doi:10.47701/djp.v3i1.2904.ac.id, doi.org.

17. Wu et al., Potential Mechanism of Curcumin and Resveratrol against SARS-CoV-2, Research Square, doi:10.21203/rs.3.rs-2780614/v1.researchsquare.com, doi.org.

18. Nag et al., Curcumin inhibits spike protein of new SARS-CoV-2 variant of concern (VOC) Omicron, an in silico study, Computers in Biology and Medicine, doi:10.1016/j.compbiomed.2022.105552.sciencedirect.com, doi.org.

19. Rampogu et al., Molecular Docking and Molecular Dynamics Simulations Discover Curcumin Analogue as a Plausible Dual Inhibitor for SARS-CoV-2, International Journal of Molecular Sciences, doi:10.3390/ijms23031771.mdpi.com, doi.org.

20. Singh (C) et al., Potential of turmeric-derived compounds against RNA-dependent RNA polymerase of SARS-CoV-2: An in-silico approach, Computers in Biology and Medicine, doi:10.1016/j.compbiomed.2021.104965.sciencedirect.com, doi.org.

21. Kandeil et al., Bioactive Polyphenolic Compounds Showing Strong Antiviral Activities against Severe Acute Respiratory Syndrome Coronavirus 2, Pathogens, doi:10.3390/pathogens10060758.mdpi.com, doi.org.

22. Rajagopal et al., Activity of phytochemical constituents of Curcuma longa (turmeric) and Andrographis paniculata against coronavirus (COVID-19): an in silico approach, Future Journal of Pharmaceutical Sciences, doi:10.1186/s43094-020-00126-x.springeropen.com, doi.org.

23. Suravajhala et al., Comparative Docking Studies on Curcumin with COVID-19 Proteins, MDPI AG, doi:10.20944/preprints202005.0439.v3.preprints.org, doi.org.

24. Sekiou et al., In-Silico Identification of Potent Inhibitors of COVID-19 Main Protease (Mpro) and Angiotensin Converting Enzyme 2 (ACE2) from Natural Products: Quercetin, Hispidulin, and Cirsimaritin Exhibited Better Potential Inhibition than Hydroxy-Chloroquine Against COVID-19 Main Protease Active Site and ACE2, ChemRxiv, doi:10.26434/chemrxiv.12181404.v1.chemrxiv.org, doi.org.

25. Li et al., Thermal shift assay (TSA)-based drug screening strategy for rapid discovery of inhibitors against the Nsp13 helicase of SARS-CoV-2, Animals and Zoonoses, doi:10.1016/j.azn.2024.06.001.sciencedirect.com, doi.org.

26. Kamble et al., Nanoparticulate curcumin spray imparts prophylactic and therapeutic properties against SARS-CoV-2, Emergent Materials, doi:10.1007/s42247-024-00754-6.springer.com, doi.org.

27. Nicoliche et al., Antiviral, anti-inflammatory and antioxidant effects of curcumin and curcuminoids in SH-SY5Y cells infected by SARS-CoV-2, Scientific Reports, doi:10.1038/s41598-024-61662-7.nature.com, doi.org.

28. Nittayananta et al., A novel film spray containing curcumin inhibits SARS-CoV-2 and influenza virus infection and enhances mucosal immunity, Virology Journal, doi:10.1186/s12985-023-02282-x.biomedcentral.com, doi.org.

29. Septisetyani et al., Curcumin and turmeric extract inhibited SARS-CoV-2 pseudovirus cell entry and Spike mediated cell fusion, bioRxiv, doi:10.1101/2023.09.28.560070.biorxiv.org, doi.org.

30. Mohd Abd Razak et al., In Vitro Anti-SARS-CoV-2 Activities of Curcumin and Selected Phenolic Compounds, Natural Product Communications, doi:10.1177/1934578X231188861.sagepub.com, doi.org.

31. Teshima et al., Antiviral activity of curcumin and its analogs selected by an artificial intelligence-supported activity prediction system in SARS-CoV-2-infected VeroE6 cells, Natural Product Research, doi:10.1080/14786419.2023.2194647.tandfonline.com, doi.org.

32. Leka et al., In vitro antiviral activity against SARS-CoV-2 of common herbal medicinal extracts and their bioactive compounds, Phytotherapy Research, doi:10.1002/ptr.7463.wiley.com, doi.org.

33. Goc et al., Inhibitory effects of specific combination of natural compounds against SARS-CoV-2 and its Alpha, Beta, Gamma, Delta, Kappa, and Mu variants, European Journal of Microbiology and Immunology, doi:10.1556/1886.2021.00022.akjournals.com, doi.org.

34. Marín-Palma et al., Curcumin Inhibits In Vitro SARS-CoV-2 Infection In Vero E6 Cells through Multiple Antiviral Mechanisms, Molecules, doi:10.3390/molecules26226900.mdpi.com, doi.org.

35. Bahun et al., Inhibition of the SARS-CoV-2 3CLpro main protease by plant polyphenols, Food Chemistry, doi:10.1016/j.foodchem.2021.131594.sciencedirect.com, doi.org.

36. Bormann et al., Turmeric Root and Its Bioactive Ingredient Curcumin Effectively Neutralize SARS-CoV-2 In Vitro, Viruses, doi:10.3390/v13101914.mdpi.com, doi.org.

37. Guijarro-Real et al., Potential In Vitro Inhibition of Selected Plant Extracts against SARS-CoV-2 Chymotripsin-Like Protease (3CLPro) Activity, Foods, doi:10.3390/foods10071503.mdpi.com, doi.org.

38. Goc (B) et al., Phenolic compounds disrupt spike-mediated receptor-binding and entry of SARS-CoV-2 pseudo-virions, PLOS ONE, doi:10.1371/journal.pone.0253489.plos.org, doi.org.

a. The trimeric spike (S) protein is a glycoprotein that mediates viral entry by binding to the host ACE2 receptor, is critical for SARS-CoV-2's ability to infect host cells, and is a target of neutralizing antibodies. Inhibition of the spike protein prevents viral attachment, halting infection at the earliest stage.

b. The receptor binding domain is a specific region of the spike protein that binds ACE2 and is a major target of neutralizing antibodies. Focusing on the precise binding site allows highly specific disruption of viral attachment with reduced potential for off-target effects.

c. The main protease or M^pro, also known as 3CL^pro or nsp5, is a cysteine protease that cleaves viral polyproteins into functional units needed for replication. Inhibiting M^pro disrupts the SARS-CoV-2 lifecycle within the host cell, preventing the creation of new copies.

d. RNA-dependent RNA polymerase (RdRp), also called nsp12, is the core enzyme of the viral replicase-transcriptase complex that copies the positive-sense viral RNA genome into negative-sense templates for progeny RNA synthesis. Inhibiting RdRp blocks viral genome replication and transcription.

e. The angiotensin converting enzyme 2 (ACE2) protein is a host cell transmembrane protein that serves as the cellular receptor for the SARS-CoV-2 spike protein. ACE2 is expressed on many cell types, including epithelial cells in the lungs, and allows the virus to enter and infect host cells. Inhibition may affect ACE2's physiological function in blood pressure control.

f. The nucleocapsid (N) protein binds and encapsulates the viral genome by coating the viral RNA. N enables formation and release of infectious virions and plays additional roles in viral replication and pathogenesis. N is also an immunodominant antigen used in diagnostic assays.

g. Non-structural protein 10 (nsp10) serves as an RNA chaperone and stabilizes conformations of nsp12 and nsp14 in the replicase-transcriptase complex, which synthesizes new viral RNAs. Nsp10 disruption may destabilize replicase-transcriptase complex activity.

h. The helicase, or nsp13, protein unwinds the double-stranded viral RNA, a crucial step in replication and transcription. Inhibition may prevent viral genome replication and the creation of new virus components.

i. Transmembrane protease serine 2 (TMPRSS2) is a host cell protease that primes the spike protein, facilitating cellular entry. TMPRSS2 activity helps enable cleavage of the spike protein required for membrane fusion and virus entry. Inhibition may especially protect respiratory epithelial cells, buy may have physiological effects.

j. Calu-3 is a human lung adenocarcinoma cell line with moderate ACE2 and TMPRSS2 expression and SARS-CoV-2 susceptibility. It provides a model of the human respiratory epithelium, but many not be ideal for modeling early stages of infection due to the moderate expression levels of ACE2 and TMPRSS2.

k. A549 is a human lung carcinoma cell line with low ACE2 expression and SARS-CoV-2 susceptibility. Viral entry/replication can be studied but the cells may not replicate all aspects of lung infection.

l. 293T is a human embryonic kidney cell line that can be engineered for high ACE2 expression and SARS-CoV-2 susceptibility. 293T cells are easily transfected and support high protein expression.

m. HEK293-hACE2 is a human embryonic kidney cell line with high ACE2 expression and SARS-CoV-2 susceptibility. Cells have been transfected with a plasmid to express the human ACE2 (hACE2) protein.

n. 293T/hACE2/TMPRSS2 is a human embryonic kidney cell line engineered for high ACE2 and TMPRSS2 expression, which mimics key aspects of human infection. 293T/hACE2/TMPRSS2 cells are very susceptible to SARS-CoV-2 infection.

o. Vero E6 is an African green monkey kidney cell line with low/no ACE2 expression and high SARS-CoV-2 susceptibility. The cell line is easy to maintain and supports robust viral replication, however the monkey origin may not accurately represent human responses.

p. SH-SY5Y is a human neuroblastoma cell line that exhibits neuronal phenotypes. It is commonly used as an in vitro model for studying neurotoxicity, neurodegenerative diseases, and neuronal differentiation.

Wu et al., 13 Apr 2023, preprint, 6 authors.

In Vitro studies are an important part of preclinical research, however results may be very different in vivo.

This Paper Curcumin All

Potential Mechanism of Curcumin and Resveratrol against SARS-CoV-2

Wei Wu, Junxi Wu, Xuxu Ji, Ji Liu, Fuchang Geng

doi:10.21203/rs.3.rs-2780614/v1

Recently, World Health Organization predicted a near end of COVID-19 pandemic. However, the prediction should be interpreted cautiously. Due to SARS-CoV-2 continuous mutation-evolve, limited durability of infection-acquired protection in individuals with hybrid immunity, and the effects of long COVID-19 or Post-COVID-19 syndrome, COVID-19 may continue to be a worldwide threat. Alternative therapeutics are incorporated into some countries' health guidelines for COVID-19. Qiannan herbal, an ancient medical book of Yi Nationality in China, recorded that grapes and turmeric were often used to treat respiratory diseases. Curcumin and resveratrol are the primary bioactive compounds in turmeric and grapes, respectively. The clinical trials con rmed that curcumin or resveratrol supplementation could cause moderate or marked improvements in COVID-19 patients. Exploring the potential mechanisms is of great signi cance. This study found that curcumin and resveratrol could effectively inhibit SARS-CoV-23CLpro activity and spike protein-mediated cell entry. Curcumin and resveratrol could signi cantly alleviate spike protein-mediated cytokine storm via inhibiting over-activation of NFKB, and effectively ameliorate spike protein-mediated oxidative stress through scavenging ROS and enhancing function of antioxidation system. The combined treatment showed a better effect than alone treatment. Therefore, curcumin and resveratrol could inhibit SARS-CoV-23C-like proteinase activity and Spike protein-mediated cell entry, cytokine storm, and oxidative stress.

In conclusion, curcumin and resveratrol could inhibit SARS-CoV-2 3CLpro activity and spike proteinmediated cell entry, cytokine storm, and oxidative stress. The above conclusions are shown in Supplementary Fig. 5 . Our study provides a reference of nutrient supplementation for preventing and treating COVID-19. Methods Venn analysis and Enrichment analysis Firstly, COVID-19, curcumin, and resveratrol were separately input into Gene or Pubchem database at National Center for Biotechnology Information (NCBI) and ltered with Homo sapiens, and the gene sets were Cell culture and reagents Vero cells and HEK293T-hACE2 cells were obtained from the Laboratory of Biochemistry and Molecular Biology, Sichuan University. Cells were maintained in Dulbecco's Modi ed Eagle's Medium (DMEM) supplemented with 10% FBS and incubated at 37°C with 5% CO2. Fetal bovine serum (FBS), DMEM, and phosphate-buffered saline (PBS) were bought from Gibco (Grand Island, NY, USA). Pyrrolidinedithiocarbamate ammonium (PDTC), curcumin, and resveratrol were obtained from Solarbio. Ebselen was purchased from Beyotime. Cell viability assay Vero E6 cells were..

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{ 'institution': [{'name': 'Research Square'}], 'indexed': {'date-parts': [[2023, 6, 29]], 'date-time': '2023-06-29T08:13:11Z', 'timestamp': 1688026391612}, 'posted': {'date-parts': [[2023, 4, 14]]}, 'group-title': 'In Review', 'reference-count': 62, 'publisher': 'Research Square Platform LLC', 'license': [ { 'start': { 'date-parts': [[2023, 4, 14]], 'date-time': '2023-04-14T00:00:00Z', 'timestamp': 1681430400000}, 'content-version': 'unspecified', 'delay-in-days': 0, 'URL': 'https://creativecommons.org/licenses/by/4.0/'}], 'content-domain': {'domain': [], 'crossmark-restriction': False}, 'accepted': {'date-parts': [[2023, 4, 5]]}, 'abstract': 'Abstract\n' ' Recently, World Health Organization predicted a near end of COVID-19 ' 'pandemic. However, the prediction should be interpreted cautiously. Due to SARS-CoV-2 ' 'continuous mutation-evolve, limited durability of infection-acquired protection in ' 'individuals with hybrid immunity, and the effects of long COVID-19 or Post-COVID-19 syndrome, ' 'COVID-19 may continue to be a worldwide threat. Alternative therapeutics are incorporated ' 'into some countries’ health guidelines for COVID-19. Qiannan herbal, an ancient medical book ' 'of Yi Nationality in China, recorded that grapes and turmeric were often used to treat ' 'respiratory diseases. Curcumin and resveratrol are the primary bioactive compounds in ' 'turmeric and grapes, respectively. The clinical trials confirmed that curcumin or resveratrol ' 'supplementation could cause moderate or marked improvements in COVID-19 patients. Exploring ' 'the potential mechanisms is of great significance. This study found that curcumin and ' 'resveratrol could effectively inhibit SARS-CoV-23CLpro activity and spike protein-mediated ' 'cell entry. Curcumin and resveratrol could significantly alleviate spike protein-mediated ' 'cytokine storm via inhibiting over-activation of NFKB, and effectively ameliorate spike ' 'protein-mediated oxidative stress through scavenging ROS and enhancing function of ' 'antioxidation system. The combined treatment showed a better effect than alone treatment. ' 'Therefore, curcumin and resveratrol could inhibit SARS-CoV-23C-like proteinase activity and ' 'Spike protein-mediated cell entry, cytokine storm, and oxidative stress.', 'DOI': '10.21203/rs.3.rs-2780614/v1', 'type': 'posted-content', 'created': {'date-parts': [[2023, 4, 14]], 'date-time': '2023-04-14T13:37:06Z', 'timestamp': 1681479426000}, 'source': 'Crossref', 'is-referenced-by-count': 0, 'title': 'Potential Mechanism of Curcumin and Resveratrol against SARS-CoV-2', 'prefix': '10.21203', 'author': [ { 'given': 'Wei', 'family': 'Wu', 'sequence': 'first', 'affiliation': [ { 'name': 'Liangshan Prefecture Integrated Traditional and Western Medicine ' 'Hospital'}]}, { 'given': 'Junxi', 'family': 'Wu', 'sequence': 'additional', 'affiliation': [{'name': 'Chengdu University of TCM'}]}, { 'given': 'Xuxu', 'family': 'Ji', 'sequence': 'additional', 'affiliation': [{'name': 'Sichuan University'}]}, { 'given': 'Ji', 'family': 'Liu', 'sequence': 'additional', 'affiliation': [{'name': 'Sichuan University'}]}, { 'given': 'Bin', 'family': 'Liu', 'sequence': 'additional', 'affiliation': [{'name': 'Good Doctor Pharmaceutical Group Co.Ltd'}]}, { 'given': 'Fuchang', 'family': 'Geng', 'sequence': 'additional', 'affiliation': [{'name': 'Good Doctor Pharmaceutical Group Co.Ltd'}]}], 'member': '8761', 'reference': [ { 'key': 'ref1', 'unstructured': 'https://www.who.int/emergencies/diseases/novel-coronavirus-2019.'}, { 'issue': '2', 'key': 'ref2', 'doi-asserted-by': 'publisher', 'first-page': '114', 'DOI': '10.37201/req/122.2022', 'article-title': 'Insights for COVID-19 in 2023', 'volume': '36', 'author': 'Martín Sánchez FJ', 'year': '2023', 'unstructured': 'Martín Sánchez FJ, et al. 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