Exploring the Potential of Siddha Formulation MilagaiKudineer-Derived Phytotherapeutics Against SARS-CoV-2: An In-Silico Investigation for Antiviral Intervention

Rajamanickam et al., Journal of Pharmacy and Pharmacology Research, doi:10.26502/fjppr.0105, Feb 2025

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Vitamin C for COVID-19

6th treatment shown to reduce risk in September 2020, now with p = 0.00000002 from 75 studies, recognized in 22 countries.

Lower risk for mortality, ICU, hospitalization, and recovery.

No treatment is 100% effective. Protocols combine treatments.

5,700+ studies for 169 treatments. c19early.org

In-Silico computational docking study of phytochemicals from MilagaiKudineer, showing potential inhibitory activity against SARS-CoV-2 targets. Authors evaluated binding affinities for the spike glycoprotein receptor-binding domain, RNA-dependent RNA polymerase, and the main protease (3CL^pro). Compounds including curcumin, quercetin, capsaicin, and dihydrocapsaicin demonstrated significant binding affinities, suggesting potential antiviral mechanisms through disruption of viral entry and replication. Vitamin C showed moderate affinity that may contribute synergistically in combination therapies.

17 preclinical studies support the efficacy of vitamin C for COVID-19:

8 In Silico studies1-8

8 In Vitro studies1,7,9-14

1 In Vivo animal study12

Vitamin C has been identified by the European Food Safety Authority (EFSA) as having sufficient evidence for a causal relationship between intake and optimal immune system function15-17. Vitamin C plays a key role in the immune system, supporting the production and function of leukocytes, or white blood cells, which defend against infection and disease, including the production of lymphocytes, which make antibodies, and enhancing phagocytosis, the process by which immune system cells ingest and destroy viruses and infected cells. Vitamin C is an antioxidant, protecting cells from damage caused by free radicals. Vitamin C inhibits SARS-CoV-2 3CL^pro7,11, inhibits SARS-CoV-2 infection by reducing ACE2 levels in a dose-dependent manner12, and may limit COVID-19 induced cardiac damage by acting as an antioxidant and potentially reducing the reactive oxygen species (ROS) production induced by the spike protein that contributes to the activation of profibrotic pathways9. Vitamin C reduces inflammation, oxidative stress, and NETosis, supporting immune function and vascular protection18. Intracellular levels of vitamin C decline during COVID-19 hospitalization suggesting ongoing utilization and depletion of vitamin C19. Threonic acid, a metabolite of vitamin C, is lower in mild and severe cases, consistent with increased need for and metabolization of vitamin C with moderate infection, but more limited ability to produce threonic acid in severe infection due to depletion or existing lower levels of vitamin C20. Symptomatic COVID-19 is associated with a lower frequency of natural killer (NK) cells, and vitamin C has been shown to improve NK cell numbers and functioning21,22.

Important missing comments or errors? Let us know.

Study covers quercetin, curcumin, and vitamin C.

1. Najimi et al., Phytochemical Inhibitors of SARS‐CoV‐2 Entry: Targeting the ACE2‐RBD Interaction with l‐Tartaric Acid, l‐Ascorbic Acid, and Curcuma longa Extract, ChemistrySelect, doi:10.1002/slct.202406035.wiley.com, doi.org.

2. Rajamanickam et al., Exploring the Potential of Siddha Formulation MilagaiKudineer-Derived Phytotherapeutics Against SARS-CoV-2: An In-Silico Investigation for Antiviral Intervention, Journal of Pharmacy and Pharmacology Research, doi:10.26502/fjppr.0105.fortunejournals.com, doi.org.

3. 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.

4. Morales-Bayuelo et al., New findings on ligand series used as SARS-CoV-2 virus inhibitors within the frameworks of molecular docking, molecular quantum similarity and chemical reactivity indices, F1000Research, doi:10.12688/f1000research.123550.3.f1000research.com, doi.org.

5. 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.

6. Pandya et al., Unravelling Vitamin B12 as a potential inhibitor against SARS-CoV-2: A computational approach, Informatics in Medicine Unlocked, doi:10.1016/j.imu.2022.100951.sciencedirect.com, doi.org.

7. Malla et al., Vitamin C inhibits SARS coronavirus-2 main protease essential for viral replication, bioRxiv, doi:10.1101/2021.05.02.442358.biorxiv.org, doi.org.

8. Kumar et al., In silico virtual screening-based study of nutraceuticals predicts the therapeutic potentials of folic acid and its derivatives against COVID-19, VirusDisease, doi:10.1007/s13337-020-00643-6.springer.com, doi.org.

9. Van Tin et al., Spike Protein of SARS-CoV-2 Activates Cardiac Fibrogenesis through NLRP3 Inflammasomes and NF-κB Signaling, Cells, doi:10.3390/cells13161331.mdpi.com, doi.org.

10. Moatasim et al., Potent Antiviral Activity of Vitamin B12 against Severe Acute Respiratory Syndrome Coronavirus 2, Middle East Respiratory Syndrome Coronavirus, and Human Coronavirus 229E, Microorganisms, doi:10.3390/microorganisms11112777.mdpi.com, doi.org.

11. Đukić et al., Inhibition of SARS-CoV-2 Mpro with Vitamin C, L-Arginine and a Vitamin C/L-Arginine Combination, Frontiers in Bioscience-Landmark, doi:10.31083/j.fbl2801008.imrpress.com, doi.org.

12. Zuo et al., Vitamin C promotes ACE2 degradation and protects against SARS‐CoV‐2 infection, EMBO reports, doi:10.15252/embr.202256374.wiley.com, doi.org.

13. Hajdrik et al., In Vitro Determination of Inhibitory Effects of Humic Substances Complexing Zn and Se on SARS-CoV-2 Virus Replication, Foods, doi:10.3390/foods11050694.mdpi.com, doi.org.

14. 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.

15. Galmés et al., Suboptimal Consumption of Relevant Immune System Micronutrients Is Associated with a Worse Impact of COVID-19 in Spanish Populations, Nutrients, doi:10.3390/nu14112254.mdpi.com, doi.org.

16. Galmés (B) et al., Current State of Evidence: Influence of Nutritional and Nutrigenetic Factors on Immunity in the COVID-19 Pandemic Framework, Nutrients, doi:10.3390/nu12092738.mdpi.com, doi.org.

17. EFSA, Scientific Opinion on the substantiation of health claims related to vitamin C and protection of DNA, proteins and lipids from oxidative damage (ID 129, 138, 143, 148), antioxidant function of lutein (ID 146), maintenance of vision (ID 141, 142), collagen formation (ID 130, 131, 136, 137, 149), function of the nervous system (ID 133), function of the immune system (ID 134), function of the immune system during and after extreme physical exercise (ID 144), non-haem iron absorption (ID 132, 147), energy-yielding metabolism (ID 135), and relief in case of irritation in the upper respiratory tract (ID 1714, 1715) pursuant to Article 13(1) of Regulation (EC) No 1924/2006, EFSA Journal, doi:10.2903/j.efsa.2009.1226.europa.eu, doi.org.

18. Xie et al., The role of reactive oxygen species in severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) infection-induced cell death, Cellular & Molecular Biology Letters, doi:10.1186/s11658-024-00659-6.biomedcentral.com, doi.org.

19. Boerenkamp et al., Low Levels of Serum and Intracellular Vitamin C in Hospitalized COVID-19 Patients, Nutrients, doi:10.3390/nu15163653.mdpi.com, doi.org.

20. Albóniga et al., Differential abundance of lipids and metabolites related to SARS-CoV-2 infection and susceptibility, Scientific Reports, doi:10.1038/s41598-023-40999-5.nature.com, doi.org.

21. Graydon et al., High baseline frequencies of natural killer cells are associated with asymptomatic SARS-CoV-2 infection, Current Research in Immunology, doi:10.1016/j.crimmu.2023.100064.sciencedirect.com, doi.org.

22. Vojdani et al., In vivo effect of ascorbic acid on enhancement of human natural killer cell activity, Nutrition Research, doi:10.1016/S0271-5317(05)80799-7.sciencedirect.com, doi.org.

Rajamanickam et al., 20 Feb 2025, India, peer-reviewed, 7 authors.

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

This Paper Vitamin C All

Exploring the Potential of Siddha Formulation MilagaiKudineer-Derived Phytotherapeutics Against SARS-CoV-2: An In-Silico Investigation for Antiviral Intervention

Lecturer II Baskar Rajamanickam, Ramesh Balasubramanian, Manekshah Yovas Rajammal, Bharathkumar Govindaraju, Selvapriya Sikkal Selvaraaju, Hemadevi Thangarasu, Kanakavalli Kadarkarai

Journal of Pharmacy and Pharmacology Research, doi:10.26502/fjppr.0105

The search for effective therapeutics against COVID-19 remains imperative, and natural compounds have emerged as promising candidates. Our study explores the potential of bioactive phytochemicals from the traditional Siddha formulation MilagaiKudineer as inhibitors against key target proteins of the SARS-CoV-2 virus. Through in-silico docking analyses, the interactions of phytochemicals from Cuminum cyminum, Curcuma longa, and Capsicum annuum with the receptor-binding domain of the SARS-CoV-2 spike glycoprotein (PDB ID: 6VSB), the SARS-CoV2 RNA-dependent RNA polymerase (PDB ID: 6NUR), and the main protease, 3CL pro (PDB ID: 6LU7) were examined. Notable compounds such as Curcumin, Quercetin, Capsaicin, and Ascorbic acid demonstrated significant binding affinities towards these viral targets, suggesting mechanisms by which these phytochemicals may disrupt viral entry and replication. Our findings also highlight the potential of compounds like Carvacrol, Cuminaldehyde, Linalool, and Dihydrocapsaicin in mediating antiviral effects by interfacing with key amino acid residues of the spike glycoprotein. These interactions are indicative of their capacity to hinder the virus-host cellbinding process. Moreover, the interaction of select phytochemicals with the SARS-CoV2 RNA-dependent RNA polymerase and the 3CLpro enzyme suggests a possible inhibitory effect on viral replication. Given the promising interactions observed, these phytochemicals warrant further investigation through in vitro and in vivo studies to validate their antiviral efficacy against COVID-19. This research underscores the importance of exploring traditional medicinal formulations for potential therapeutic agents in the fight against emerging infectious diseases.

Compounds Conclusion MilagaiKudineer exhibits promising antiviral properties attributed to its constituents. Capsicumannuum Linn (Green chilly) contains capsaicin, which possesses antiviral effects. Cuminum cyminum Linn (Cumin) and Curcuma longa Linn (Turmeric) are known for theirimmunomodulatory and anti-inflammatory properties, which can help combat viral infections. Computational analysis revealed the potential of bioactive compounds from MilagaiKudineer to bind effectively to the receptor-binding domain of the SARS-CoV-2 spike glycoprotein, the SARS-CoV-2 RNA-dependent RNA polymerase and the SARS-CoV-2 Main protease. Compounds such as Carvacrol, Cuminaldehyde, Linalool, Curcumin, Quercetin, Cineol, Dihydrocapsaicin, Capsaicin, and Ascorbic acid hold promise as inhibitors of viral entry and replication, highlighting their potential as therapeutic agents against COVID-19. Furtherexperimental studies are warranted to validate these findings and explore their clinical applications.

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