METHYLENE BLUE INHIBITS THE INTERACTION BETWEEN HEPARAN SULFATE AND SARS-COV-2 SPIKE PROTEIN; A REVIEW OF EVIDENCE FOR A HYPOTHESIS.
DOI:
https://doi.org/10.31674/mjmr.2021.v05i04.002Abstract
The addition of methylene blue to the standard treatment protocol has been shown to improve respiratory rate and oxygen saturation in COVID-19 patients, reducing morbidity and mortality. Evidence to date suggests that methylene blue inhibits protein-protein interactions between SARS-CoV-2 Spike protein and angiotensin-converting enzyme 2, which in turn inhibits the cell entry of SARS-CoV-2. However, the methylene blue dye-binding characteristics of sulfated glycosaminoglycans suggest additional inhibitory effects of the spike protein-heparan sulfate interaction. We hypothesize that the binding of cationic methylene blue neutralizes polyanionic heparan sulfate molecules on the host cell surface. As a consequence, electrostatic interactions between negatively charged heparan sulfate and the positively charged receptor binding domain of SARS-CoV-2 spike protein will be inhibited. Thus, methylene blue will exhibit a "shielding effect" on the heparan sulfate proteoglycans, inhibiting viral attachment to the cell surface. The proposed mechanism corroborates the possible broad-spectrum antiviral activity of methylene blue against multiple human coronaviruses that exploit the electrostatic interactions with sulfated glycosaminoglycans for virus entry. Methylene blue would exhibit the same anti-adhesive activity at the blood-brain-barrier and olfactory neuroepithelium, corroborating potential benefits in ameliorating post-COVID-19 neurological impairment. However, as cationic dyes can bind to both free glycosaminoglycans in circulation as well as proteoglycans attached to the cell surface, co-administration of intravenous heparin could possibly antagonize the proposed antiviral activity. This critical review focuses on empirical evidence to support the hypothesized heparan sulfate-dependent antiviral activity of MB.
Keywords:
Methylene blue, SARS-CoV-2, COVID-19, Heparan sulfate, Proteoglycans, AntiviralDownloads
References
Buzon, M. J., Seiss, K., Weiss, R., Brass, A. L., Rosenberg, E. S., Pereyra, F., ... & Lichterfeld, M. (2011). Inhibition of HIV-1 integration in ex vivo-infected CD4 T cells from elite controllers. Journal of virology, 85(18), 9646-9650.
Bojadzic, D., Alcazar, O., & Buchwald, P. (2021). Methylene blue inhibits the SARS-CoV-2 spike–ACE2 protein-protein interaction–a mechanism that can contribute to its antiviral activity against COVID-19. Frontiers in pharmacology, 2255.
Bojadzic, D., Alcazar, O., Chen, J., Chuang, S. T., Condor Capcha, J. M., Shehadeh, L. A., & Buchwald, P. (2021). Small-Molecule Inhibitors of the Coronavirus Spike: ACE2 Protein–Protein Interaction as Blockers of Viral Attachment and Entry for SARS-CoV-2. ACS Infectious Diseases.
Buijsers, B., Yanginlar, C., Maciej-Hulme, M. L., de Mast, Q., & van der Vlag, J. (2020). Beneficial non-anticoagulant mechanisms underlying heparin treatment of COVID-19 patients. EBioMedicine, 59, 102969.
Cagno, Valeria, Chiara Medaglia, Andreas Cerny, Thomas Cerny, Caroline Tapparel, and Erich Cerny. "Methylene blue has a potent antiviral activity against SARS-CoV-2 in the absence of UV-activation in vitro." bioRxiv (2020).
Chu, H., Hu, B., Huang, X., Chai, Y., Zhou, D., Wang, Y., ... & Yuen, K. Y. (2021). Host and viral determinants for efficient SARS-CoV-2 infection of the human lung. Nature communications, 12(1), 1-15.
Clausen, T. M., Sandoval, D. R., Spliid, C. B., Pihl, J., Perrett, H. R., Painter, C. D., ... & Esko, J. D. (2020). SARS-CoV-2 Infection Depends on Cellular Heparan Sulfate and ACE2. Cell. https. doi. org/10.1016/j. cell, 33.
Dey, P. (2018). Staining Principle and general procedure of staining of the tissue. In Basic and Advanced Laboratory Techniques in Histopathology and Cytology (pp. 57-67). Springer, Singapore.
Gasbarri, M., V’kovski, P., Torriani, G., Thiel, V., Stellacci, F., Tapparel, C., & Cagno, V. (2020). Sars-cov-2 inhibition by sulfonated compounds. Microorganisms, 8(12), 1894.
Gendrot, M., Andreani, J., Duflot, I., Boxberger, M., Le Bideau, M., Mosnier, J., ... & Pradines, B. (2020). Methylene blue inhibits replication of SARS-CoV-2 in vitro. International Journal of Antimicrobial Agents, 56(6), 106202.
Gendrot, M., Jardot, P., Delandre, O., Boxberger, M., Andreani, J., Duflot, I., ... & Pradines, B. (2021). In Vitro Evaluation of the Antiviral Activity of Methylene Blue Alone or in Combination against SARS-CoV-2. Journal of Clinical Medicine, 10(14), 3007.
Ginimuge, P. R., & Jyothi, S. (2010). Methylene blue: revisited. Journal of anaesthesiology, clinical pharmacology, 26(4), 517.
Hamidi-Alamdari, D., Hafizi-Lotfabadi, S., Bagheri-Moghaddam, A., Safari, H., Mozdourian, M., Javidarabshahi, Z., ... & Koliakos, G. (2021). Methylene blue for treatment of hospitalized COVID-19 patients: A randomized, controlled, open-label clinical trial, phase 2. Revista de investigación clínica, 73(3), 190-198.
Hao, W., Ma, B., Li, Z., Wang, X., Gao, X., Li, Y., ... & Tan, Z. (2021). Binding of the SARS-CoV-2 spike protein to glycans. Science Bulletin, 66(12), 1205-1214.
Hassanzadeh, K., Perez Pena, H., Dragotto, J., Buccarello, L., Iorio, F., Pieraccini, S., ... & Feligioni, M. (2020). Considerations around the SARS-CoV-2 Spike Protein with particular attention to COVID-19 brain infection and neurological symptoms. ACS chemical neuroscience, 11(15), 2361-2369.
Hu, Y., Meng, X., Zhang, F., Xiang, Y., & Wang, J. (2021). The in vitro antiviral activity of lactoferrin against common human coronaviruses and SARS-CoV-2 is mediated by targeting the heparan sulfate co-receptor. Emerging microbes & infections, 10(1), 317-330.
Hudák, A., Letoha, A., Szilák, L., & Letoha, T. (2021). Contribution of syndecans to the cellular entry of SARS-CoV-2. International journal of molecular sciences, 22(10), 5336.
Hudák, A., Letoha, A., Szilák, L., & Letoha, T. (2021). Contribution of syndecans to the cellular entry of SARS-CoV-2. International journal of molecular sciences, 22(10), 5336.
Jiao, Q., & Liu, Q. (1999). Characterization of the interaction between methylene blue and glycosaminoglycans. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 55(7-8), 1667-1673.
Jin, W., Zhang, W., Mitra, D., McCandless, M. G., Sharma, P., Tandon, R., ... & Linhardt, R. J. (2020). The structure-activity relationship of the interactions of SARS-CoV-2 spike glycoproteins with glucuronomannan and sulfated galactofucan from Saccharina japonica. International journal of biological macromolecules, 163, 1649-1658.
Kim, S. Y., Jin, W., Sood, A., Montgomery, D. W., Grant, O. C., Fuster, M. M., ... & Linhardt, R. J. (2020). Characterization of heparin and severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) spike glycoprotein binding interactions. Antiviral research, 181, 104873.
Koganti, R., Memon, A., & Shukla, D. (2021, April). Emerging roles of heparan sulfate proteoglycans in viral pathogenesis. In Seminars in Thrombosis and Hemostasis (Vol. 47, No. 03, pp. 283-294). Thieme Medical Publishers, Inc..
Kostin, A. I., Lundgren, M. N., Bulanov, A. Y., Ladygina, E. A., Chirkova, K. S., Gintsburg, A. L., ... & Petrikov, S. S. (2021). Impact of pathogen reduction methods on immunological properties of the COVID‐19 convalescent plasma. Vox Sanguinis.
Kumar, A., Pareek, V., Prasoon, P., Faiq, M. A., Kumar, P., Kumari, C., & Narayan, R. K. (2020). Possible routes of SARS‐CoV‐2 invasion in brain: in context of neurological symptoms in COVID‐19 patients. Journal of neuroscience research, 98(12), 2376-2383.
Kwon, P. S., Oh, H., Kwon, S. J., Jin, W., Zhang, F., Fraser, K., ... & Dordick, J. S. (2020). Sulfated polysaccharides effectively inhibit SARS-CoV-2 in vitro. Cell discovery, 6(1), 1-4.
Lang, J., Yang, N., Deng, J., Liu, K., Yang, P., Zhang, G., & Jiang, C. (2011). Inhibition of SARS pseudovirus cell entry by lactoferrin binding to heparan sulfate proteoglycans. PloS one, 6(8), e23710.
Lendvay, T. S., Chen, J., Harcourt, B. H., Scholte, F. E., Lin, Y. L., Kilinc-Balci, F. S., ... & Chu, M. C. (2020). Addressing Personal Protective Equipment (PPE) Decontamination: Methylene Blue and Light Inactivates SARS-CoV-2 on N95 Respirators and Medical Masks with Maintenance of Integrity and Fit. Infection Control & Hospital Epidemiology, 1-83.
Liu, L., Chopra, P., Li, X., Bouwman, K. M., Tompkins, S. M., Wolfert, M. A., ... & Boons, G. J. (2021). Heparan sulfate proteoglycans as attachment factor for SARS-CoV-2. ACS Central Science.
Magoon, R., Bansal, N., Singh, A., & Kashav, R. (2021). Methylene blue: Subduing the post COVID-19 blues!. Medical Hypotheses.
Milewska, A., Zarebski, M., Nowak, P., Stozek, K., Potempa, J., & Pyrc, K. (2014). Human coronavirus NL63 utilizes heparan sulfate proteoglycans for attachment to target cells. Journal of virology, 88(22), 13221-13230.
Milho, R., Frederico, B., Efstathiou, S., & Stevenson, P. G. (2012). A heparan-dependent herpesvirus targets the olfactory neuroepithelium for host entry. PLoS pathogens, 8(11), e1002986
Mycroft-West, C. J., Su, D., Pagani, I., Rudd, T. R., Elli, S., Guimond, S. E., ... & Skidmore, M. A. (2020). Heparin inhibits cellular invasion by SARS-CoV-2: structural dependence of the interaction of the surface protein (spike) S1 receptor binding domain with heparin. BioRxiv.
Németh-Csóka, M., Kajtár, J., & Kajtár, M. (1975). Biological significance of helical conformation of acid polysaccharides. Connective tissue research, 3(4), 207-211.
Andreu, G. L. P. (2021). The rationale for methylene blue utility against SARS-CoV-2 infection complications. Journal of Pharmacy & Pharmacognosy Research, 9(3), 379-396.
Reis, C. A., Tauber, R., & Blanchard, V. (2021). Glycosylation is a key in SARS-CoV-2 infection. Journal of Molecular Medicine, 1-9.
Schuurs, Z. P., Hammond, E., Elli, S., Rudd, T. R., Mycroft-West, C. J., Lima, M. A., ... & Gandhi, N. S. (2021). Evidence of a putative glycosaminoglycan binding site on the glycosylated SARS-CoV-2 spike protein N-terminal domain. Computational and Structural Biotechnology Journal, 19, 2806-2818.
Scigliano, G., & Scigliano, G. A. (2021). Methylene blue in covid-19. Medical Hypotheses, 146, 110455.
Soares da Costa, D., Reis, R. L., & Pashkuleva, I. (2017). Sulfation of glycosaminoglycans and its implications in human health and disorders. Annual review of biomedical engineering, 19, 1-26.
Stone, J. E., Akhtar, N., Botchway, S., & Pennock, C. A. (1994). Interaction of 1, 9-dimethylmethylene blue with glycosaminoglycans. Annals of clinical biochemistry, 31(2), 147-152.
Sun, Y., & Ho, M. (2020). Emerging antibody-based therapeutics against SARS-CoV-2 during the global pandemic. Antibody therapeutics, 3(4), 246-256.
Suryawanshi, R. K., Patil, C. D., Koganti, R., Singh, S. K., Ames, J. M., & Shukla, D. (2021). Heparan Sulfate Binding Cationic Peptides Restrict SARS-CoV-2 Entry. Pathogens, 10(7), 803.
Svyatchenko, V. A., Nikonov, S. D., Mayorov, A. P., Gelfond, M. L., & Loktev, V. B. (2021). Antiviral photodynamic therapy: Inactivation and inhibition of SARS-CoV-2 in vitro using methylene blue and Radachlorin. Photodiagnosis and Photodynamic Therapy, 33, 102112.
Tandon, R., Sharp, J. S., Zhang, F., Pomin, V. H., Ashpole, N. M., Mitra, D., ... & Linhardt, R. J. (2020). Effective inhibition of SARS-CoV-2 entry by heparin and enoxaparin derivatives. Journal of virology, 95(3), e01987-20.
Tavassoly, O., Safavi, F., & Tavassoly, I. (2020). Heparin-binding Peptides as Novel Therapies to Stop SARS-CoV-2 Cellular Entry and Infection. Molecular Pharmacology, 98(5), 612-619.
Templeton, D. M. (1988). The basis and applicability of the dimethylmethylene blue binding assay for sulfated glycosaminoglycans. Connective Tissue Research, 17(1), 23-32.
Wells, L., Vierra, C., Hardman, J., Han, Y., Dimas, D., Gwarada‐Phillips, L. N., ... & McReynolds, K. D. (2021). Sulfoglycodendrimer Therapeutics for HIV‐1 and SARS‐CoV‐2. Advanced therapeutics, 4(4), 2000210..
Yan, L., Song, Y., Xia, K., He, P., Zhang, F., Chen, S., ... & Linhardt, R. J. (2021). Heparan sulfates from bat and human lung and their binding to the spike protein of SARS-CoV-2 virus. Carbohydrate Polymers, 260, 117797.
Yu, M., Zhang, T., Zhang, W., Sun, Q., Li, H., & Li, J. P. (2020). Elucidating the interactions between heparin/heparan sulfate and SARS-CoV-2-related proteins—An important strategy for developing novel therapeutics for the COVID-19 pandemic. Frontiers in Molecular Biosciences, 7.
Yue, J., Jin, W., Yang, H., Faulkner, J., Song, X., Qiu, H., ... & Wang, L. (2021). Heparan sulfate facilitates spike protein-mediated SARS-CoV-2 host cell invasion and contributes to increased infection of SARS-CoV-2 G614 mutant and in lung cancer. Frontiers in Molecular Biosciences, 8.
Zhang, L., Li, N., & Zhao, F. (2004). Spectroscopic study on the interaction between methylene blue and chondroitin 4-sulfate and its analytical application. Analytical sciences, 20(3), 445-450.
Zhang, Q., Chen, C. Z., Swaroop, M., Xu, M., Wang, L., Lee, J., . . . Ye, Y. (2020). Heparan sulfate assists SARS-CoV-2 in cell entry and can be targeted by approved drugs in vitro. Cell Discov, 6(1), 80.
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