Proteomic Analysis Identifies the Glutathione Synthesizing Enzyme Gclc as an Andrographolide Target and a Protective Factor Against Sars-Cov-2 Infection

Chaopreecha et al., Elsevier BV, doi:10.2139/ssrn.4876852

Jul 2024

Source PDF All Studies Meta AnalysisMeta

In Vitro study showing andrographolide attenuates infection of SARS-CoV-2 wildtype and omicron variants in human lung epithelial cells and monkey kidney cells. Proteomic analysis revealed andrographolide induces expression of the glutathione synthesizing enzyme GCLC and the anti-oxidative stress response in infected cells. Ectopic over-expression of GCLC or treatment with the glutathione precursor N-acetylcysteine decreased SARS-CoV-2 infection, suggesting the glutathione pathway mediates the antiviral effects of andrographolide.

22 preclinical studies support the efficacy of andrographolide for COVID-19:

12 In Silico studies1-12

8 In Vitro studies1,13-19

2 In Vivo animal studies16,19

Important missing comments or errors? Let us know.

Study covers andrographolide and N-acetylcysteine.

1. Zhang et al., Effects and Mechanisms of Andrographolide for COVID-19: A Network Pharmacology-Based and Experimentally Validated Study, Natural Product Communications, doi:10.1177/1934578X241288428.sagepub.com, doi.org.

2. Thomas et al., Cheminformatics approach to identify andrographolide derivatives as dual inhibitors of methyltransferases (nsp14 and nsp16) of SARS-CoV-2, Scientific Reports, doi:10.1038/s41598-024-58532-7.nature.com, doi.org.

3. Bhattarai et al., Investigating the binding affinity of andrographolide against human SARS-CoV-2 spike receptor-binding domain through docking and molecular dynamics simulations, Journal of Biomolecular Structure and Dynamics, doi:10.1080/07391102.2023.2174596.tandfonline.com, doi.org.

4. Nguyen et al., The Potential of Ameliorating COVID-19 and Sequelae From Andrographis paniculata via Bioinformatics, Bioinformatics and Biology Insights, doi:10.1177/11779322221149622.sagepub.com, doi.org.

5. Dassanayake et al., Molecular Docking and In-Silico Analysis of Natural Biomolecules against Dengue, Ebola, Zika, SARS-CoV-2 Variants of Concern and Monkeypox Virus, International Journal of Molecular Sciences, doi:10.3390/ijms231911131.mdpi.com, doi.org.

6. Ningrum et al., Potency Of Andrographolide, L-Mimosine And Asiaticoside Compound As Antiviral For Covid-19 Based On In Silico Method, Proceedings Universitas Muhammadiyah Yogyakarta Undergraduate Conference, doi:10.18196/umygrace.v2i2.418.ac.id, doi.org.

7. Ravichandran et al., Identification of Potential Semisynthetic Andrographolide Derivatives to Combat COVID-19 by Targeting the SARS-COV-2 Spike Protein and Human ACE2 Receptor– An In-silico Approach, Biointerface Research in Applied Chemistry, doi:10.33263/BRIAC132.155.biointerfaceresearch.com, doi.org.

8. Saeheng et al., In Silico Prediction of Andrographolide Dosage Regimens for COVID-19 Treatment, The American Journal of Chinese Medicine, doi:10.1142/S0192415X22500732.worldscientific.com, doi.org.

9. Khanal et al., Combination of system biology to probe the anti-viral activity of andrographolide and its derivative against COVID-19, RSC Advances, doi:10.1039/D0RA10529E.rsc.org, doi.org.

10. Rehan et al., A Computational Approach Identified Andrographolide as a Potential Drug for Suppressing COVID-19-Induced Cytokine Storm, Frontiers in Immunology, doi:10.3389/fimmu.2021.648250.frontiersin.org, doi.org.

11. 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.

12. Dey et al., The role of andrographolide and its derivative in COVID-19 associated proteins and immune system, Research Square, doi:10.21203/rs.3.rs-35800/v1.researchsquare.com, doi.org.

13. Chaopreecha et al., Proteomic Analysis Identifies the Glutathione Synthesizing Enzyme Gclc as an Andrographolide Target and a Protective Factor Against Sars-Cov-2 Infection, Elsevier BV, doi:10.2139/ssrn.4876852.ssrn.com, doi.org.

14. Li et al., Andrographolide suppresses SARS-CoV-2 infection by downregulating ACE2 expression: A mechanistic study, Antiviral Therapy, doi:10.1177/13596535241259952.sagepub.com, doi.org.

15. Low et al., The wide spectrum anti-inflammatory activity of andrographolide in comparison to NSAIDs: a promising therapeutic compound against the cytokine storm, bioRxiv, doi:10.1101/2024.02.21.581396.biorxiv.org, doi.org.

16. Wan et al., Synergistic inhibition effects of andrographolide and baicalin on coronavirus mechanisms by downregulation of ACE2 protein level, Scientific Reports, doi:10.1038/s41598-024-54722-5.nature.com, doi.org.

17. Siridechakorn et al., Inhibitory efficiency of Andrographis paniculata extract on viral multiplication and nitric oxide production, Scientific Reports, doi:10.1038/s41598-023-46249-y.nature.com, doi.org.

18. 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.

19. Pu et al., The effects and mechanisms of the anti-COVID-19 traditional Chinese medicine, Dehydroandrographolide from Andrographis paniculata (Burm.f.) Wall, on acute lung injury by the inhibition of NLRP3-mediated pyroptosis, Phytomedicine, doi:10.1016/j.phymed.2023.154753.sciencedirect.com, doi.org.

Chaopreecha et al., 1 Jul 2024, preprint, 9 authors. Contact: arunee.thi@mahidol.edu (corresponding author), patompon.won@mahidol.ac.th.

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

This Paper Andrographol..All

Proteomic analysis identifies the glutathione synthesizing enzyme GCLC as an andrographolide target and a protective factor against SARS-CoV-2 infection

Jarinya Chaopreecha, Nut Phueakphud, Ampa Suksatu, Sucheewin Krobthong, Suwimon Manopwisedjaroen, Nattawadee Panyain, Suradej Hongeng, Arunee Thitithanyanont, Patompon Wongtrakoongate

Even though advanced progresses have been made for COVID-19 vaccines, rapid and extensive mutations of the SARS-CoV-2 genomes have provided a selective advantage for the virus to escape the adaptive immunity leading to a critical challenge for current treatments and preventions of COVID-19. We report here the antiviral activity of andrographolide against SARS-CoV-2 wildtype and Omicron variants. Proteomic analysis was employed to identify cellular pathways and key proteins controlled by andrographolide in the human lung epithelial cells Calu-3 infected by SARS-CoV-2. Gene ontology analysis indicates that proteins involved in NRF2-regulated pathways are differentially expressed by andrographolide. Notably, andrographolide increases expression and nuclear localization of the transcription factor NRF2. In addition, transcriptional expression of GCLC and GCLM, which are NRF2 target genes, are induced by andrographolide. We further find that infection of SARS-CoV-2 results in a reduction of glutathione level in Calu-3; the effect that is rescued by andrographolide. Moreover, andrographolide also induces expression of the glutathione producing enzyme GCLC in SARS-CoV-2 infected lung epithelial cells. Importantly, an ectopic over-expression of GCLC or treatment of N-acetylcysteine in Calu-3 cells led to a decrease in SARS-CoV-2 infection. Collectively, our findings suggest the interplay between GCLC-mediated glutathione biogenesis induced by andrographolide and the anti-SARS-CoV-2 activity. The glutathione biogenesis and recycling pathways should be further exploited as a targeted therapy against SARS-CoV-2 infection.

Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. This preprint research paper has not been peer reviewed. Electronic copy available at: https://ssrn.com/abstract=4876852 P r e p r i n t n o t p e e r r e v i e w e d This preprint research paper has not been peer reviewed. Electronic copy available at: https://ssrn.com/abstract=4876852 P r e p r i n t n o t p e e r r e v i e w e d Supplementary Table 1 . Pathway analysis of up-regulated proteins upon andrographolide treatment in Calu-3 infected with SARS-CoV-2 versus to SARS-CoV-2 infection as shown in Figure 3D . The analysis was conducted using Reactome analysis. Pathway

References

Bartolini, Stabile, Bastianelli, Giustarini, Pierucci et al., SARS-CoV2 infection impairs the metabolism and redox function of cellular glutathione, Redox Biol, doi:10.1016/j.redox.2021.102041

Burgos, Alarcon, Quiroga, Manosalva, Hancke, Andrographolide, an Anti-Inflammatory Multitarget Drug: All Roads Lead to Cellular Metabolism, Molecules, doi:10.3390/molecules26010005

Che, Zhou, Liu, Xie, Liu, Andrographolide exerts anti-respiratory syncytial virus activity by up-regulating heme oxygenase-1 independent of interferon responses in human airway epithelial cells, Mol Biol Rep, doi:10.1007/s11033-023-08346-z

Dai, Chen, Chai, Zhao, Wang et al., Overview of pharmacological activities of Andrographis paniculata and its major compound andrographolide, Crit Rev Food Sci Nutr, doi:10.1080/10408398.2018.1501657

Das, Das, Swain, Mukherjee, Bhattacharya, Andrographolide induces anti-SARS-CoV-2 response through host-directed mechanism: an in silico study, Future Virol, doi:10.2217/fvl-2021-0171

Ding, Li, Song, Xiao, Huang et al., Andrographolide prevents high-fat diet-induced obesity in C57BL/6 mice by suppressing the sterol regulatory element-binding protein pathway, J Pharmacol Exp Ther, doi:10.1124/jpet.114.217968

Elumalai, Suresh Kumar, Identification of neo-andrographolide compound targeting NS1 Lys14: an important residue in NS1 activity driving dengue pathogenesis, J Biomol Struct Dyn, doi:10.1080/07391102.2022.2068073

Enmozhi, Raja, Sebastine, Joseph, Andrographolide as a potential inhibitor of SARS-CoV-2 main protease: an in silico approach, J Biomol Struct Dyn, doi:10.1080/07391102.2020.1760136

Hati, Bhattacharyya, Impact of Thiol-Disulfide Balance on the Binding of Covid-19 Spike Protein with Angiotensin-Converting Enzyme 2 Receptor, ACS Omega, doi:10.1021/acsomega.0c02125

Hillary, Ceasar, An update on COVID-19: SARS-CoV-2 variants, antiviral drugs, and vaccines, Heliyon, doi:10.1016/j.heliyon.2023.e13952

Hosakote, Liu, Castro, Garofalo, Casola, Respiratory syncytial virus induces oxidative stress by modulating antioxidant enzymes, Am J Respir Cell Mol Biol, doi:10.1165/rcmb.2008-0330OC

Hossain, Urbi, Karuniawati, Mohiuddin, Moh Qrimida et al., Andrographis paniculata (Burm. f.) Wall. ex Nees: An Updated Review of Phytochemistry, Antimicrobial Pharmacology, and Clinical Safety and Efficacy, Life, doi:10.3390/life11040348

Ji, Shen, Jiang, Morahan, Wang, Critical roles of cellular glutathione homeostasis and jnk activation in andrographolide-mediated apoptotic cell death in human hepatoma cells, Mol Carcinog, doi:10.1002/mc.20741

Jiang, Sheng, Zhang, Ma, Gao et al., Andrographis paniculata (Burm.f.) Nees and its major constituent andrographolide as potential antiviral agents, J Ethnopharmacol, doi:10.1016/j.jep.2021.113954

Jiaqi, Siqing, Qin, Di, Bei et al., Andrographolide promoted ferroptosis to repress the development of non-small cell lung cancer through activation of the mitochondrial dysfunction, Phytomedicine, doi:10.1016/j.phymed.2022.154601

Kanjanasirirat, Suksatu, Manopwisedjaroen, Munyoo, Tuchinda et al., High-content screening of Thai medicinal plants reveals Boesenbergia rotunda extract and its component Panduratin A as anti-SARS-CoV-2 agents, Scientific reports

Kongsomros, Boonyarattanasoonthorn, Phongphaew, Kasorndorkbua, Sunyakumthorn et al., In vivo evaluation of Andrographis paniculata and Boesenbergia rotunda extract activity against SARS-CoV-2 Delta variant in Golden Syrian hamsters: Potential herbal alternative for COVID-19 treatment, Journal of Traditional and Complementary Medicine, doi:10.1016/j.jtcme.2024.05.004

Krobthong, Yingchutrakul, Samutrtai, Hitakarun, Siripattanapipong et al., Utilizing Quantitative Proteomics to Identify Species-Specific Protein Therapeutic Targets for the Treatment of Leishmaniasis, ACS Omega, doi:10.1021/acsomega.1c05792

Li, Dong, Liu, Chen, Shi et al., Astaxanthin protects ARPE-19 cells from oxidative stress via upregulation of Nrf2-regulated phase II enzymes through activation of PI3K/Akt, Molecular Vision

Li, Yuan, Wu, Zhen, Sun et al., Andrographolide, a natural antiinflammatory agent: An Update, Front Pharmacol, doi:10.3389/fphar.2022.920435

Liu, Wu, Bing, Qi, Zhu et al., H1N1 influenza virus infection through NRF2-KEAP1-GCLC pathway induces ferroptosis in nasal mucosal epithelial cells, Free Radic Biol Med, doi:10.1016/j.freeradbiomed.2023.05.004

Lu, Yang, Li, Liu, Lii et al., Andrographolide inhibits TNFαinduced ICAM-1 expression via suppression of NADPH oxidase activation and induction of HO-1 and GCLM expression through the PI3K/Akt/Nrf2 and PI3K/Akt/AP-1 pathways in human endothelial cells, Biochemical Pharmacology, doi:10.1016/j.bcp.2014.06.024

Malat, Ekalaksananan, Heawchaiyaphum, Suebsasana, Roytrakul et al., Andrographolide Inhibits Epstein-Barr Virus Lytic Reactivation in EBV-Positive Cancer Cell Lines through the Modulation of Epigenetic-Related Proteins, Molecules, doi:10.3390/molecules27144666

Mir, Kapur, Singh, Sonpavde, Lillard et al., Andrographolide inhibits prostate cancer by targeting cell cycle regulators, CXCR3 and CXCR7 chemokine receptors, Cell Cycle, doi:10.1080/15384101.2016.1148836

Murae, Shimizu, Yamamoto, Kobayashi, Houri et al., The function of SARS-CoV-2 spike protein is impaired by disulfidebond disruption with mutation at cysteine-488 and by thiol-reactive N-acetyl-cysteine and glutathione, Biochem Biophys Res Commun, doi:10.1016/j.bbrc.2022.01.106

Nutho, Wilasluck, Deetanya, Wangkanont, Arsakhant et al., Discovery of C-12 dithiocarbamate andrographolide analogues as inhibitors of SARS-CoV-2 main protease: In vitro and in silico studies, Comput Struct Biotechnol J, doi:10.1016/j.csbj.2022.05.053

Panraksa, Ramphan, Khongwichit, Smith, Activity of andrographolide against dengue virus, Antiviral Res, doi:10.1016/j.antiviral.2016.12.014

Poe, Corn, N-Acetylcysteine: A potential therapeutic agent for SARS-CoV-2, Med Hypotheses, doi:10.1016/j.mehy.2020.109862

Rehan, Ahmed, Howladar, Refai, Baeissa et al., A Computational Approach Identified Andrographolide as a Potential Drug for Suppressing COVID-19-Induced Cytokine Storm, Front Immunol, doi:10.3389/fimmu.2021.648250

Sa-Ngiamsuntorn, Suksatu, Pewkliang, Thongsri, Kanjanasirirat et al., Anti-SARS-CoV-2 Activity of Andrographis paniculata Extract and Its Major Component Andrographolide in Human Lung Epithelial Cells and Cytotoxicity Evaluation in Major Organ Cell Representatives, J Nat Prod, doi:10.1021/acs.jnatprod.0c01324

Saliu, Umar, Ogunsile, Okpara, Yanaka et al., Molecular docking and pharmacokinetic studies of phytocompounds from Nigerian Medicinal Plants as promising inhibitory agents against SARS-CoV-2 methyltransferase (nsp16), J Genet Eng Biotechnol, doi:10.1186/s43141-021-00273-5

Saravolatz, Depcinski, Sharma, Molnupiravir and Nirmatrelvir-Ritonavir: Oral Coronavirus Disease 2019 Antiviral Drugs, Clinical Infectious Diseases, doi:10.1093/cid/ciac180

Schulte, Konig, Escher, Wittenburg, Proj et al., Andrographolide Derivatives Target the KEAP1/NRF2 Axis and Possess Potent Anti-SARS-CoV-2 Activity, ChemMedChem, doi:10.1002/cmdc.202100732

Shrestha, Foster, Rawlinson, Tedla, Bull, Evolution of the SARS-CoV-2 omicron variants BA.1 to BA.5: Implications for immune escape and transmission, Rev Med Virol, doi:10.1002/rmv.2381

Soto, Manzano-Pech, Palacios-Chavarria, Valdez-Vazquez, Guarner-Lans et al., N-Acetyl Cysteine Restores the Diminished Activity of the Antioxidant Enzymatic System Caused by SARS-CoV-2 Infection: Preliminary Findings, Pharmaceuticals, doi:10.3390/ph16040591

Srikanth, Sarma, Andrographolide binds to spike glycoprotein and RNA-dependent RNA polymerase (NSP12) of SARS-CoV-2 by in silico approach: a probable molecule in the development of anti-coronaviral drug, J Genet Eng Biotechnol, doi:10.1186/s43141-021-00201-7

Tavassolifar, Aghdaei, Sadatpour, Maleknia, Fayazzadeh et al., New insights into extracellular and intracellular redox status in COVID-19 patients, Redox Biol, doi:10.1016/j.redox.2022.102563

Tian, Chen, Feng, Nirmatrelvir-ritonavir compared with other antiviral drugs for the treatment of COVID-19 patients: A systematic review and meta-analysis, J Med Virol, doi:10.1002/jmv.28732

Valko, Leibfritz, Moncol, Cronin, Mazur et al., Free radicals and antioxidants in normal physiological functions and human disease, Int J Biochem Cell Biol, doi:10.1016/j.biocel.2006.07.001

Vetvicka, Vannucci, Biological properties of andrographolide, an active ingredient of Andrographis Paniculata: a narrative review, Ann Transl Med, doi:10.21037/atm-20-7830

Vitiello, Sars-Cov-2 and risk of antiviral drug resistance, Ir J Med Sci, doi:10.1007/s11845-021-02820-y

Wang, Li, Chen, Liu, Cui et al., Identification of PDCD2 as a Candidate Target of Andrographolide That Arrests the Tumor Cell Cycle by Human Proteome-Scale Screening, ACS Pharmacol Transl Sci, doi:10.1021/acsptsci.2c00092

Who, World health organization (WHO) coronavirus (COVID-19) dashboard

Wintachai, Kaur, Lee, Ramphan, Kuadkitkan et al., Activity of andrographolide against chikungunya virus infection, Sci Rep, doi:10.1038/srep14179

Wong, Ng, Leung, Boh, Lim et al., Regulation of the NRF2 transcription factor by andrographolide and organic extracts from plant endophytes, PLoS One, doi:10.1371/journal.pone.0204853

Woo, Waye, Tsui, Yeung, Cheng, Andrographolide up-regulates cellular-reduced glutathione level and protects cardiomyocytes against hypoxia/reoxygenation injury, J Pharmacol Exp Ther, doi:10.1124/jpet.107.133918

Wroblewska, Wroblewski, Holynska-Iwan, Modrzejewska, Nuszkiewicz et al., The Role of Glutathione in Selected Viral Diseases, Antioxidants, doi:10.3390/antiox12071325

Yu, Dai, Jiang, Li, Chen et al., Andrographolide as an anti-H1N1 drug and the mechanism related to retinoic acidinducible gene-I-like receptors signaling pathway, Chin J Integr Med, doi:10.1007/s11655-014-1860-0

Zhang, Ding, Cheng, Zhang, Xiang et al., Andrographolide attenuates oxidative stress injury in cigarette smoke extract exposed macrophages through inhibiting SIRT1/ERK signaling, Int Immunopharmacol, doi:10.1016/j.intimp.2020.106230

{ 'indexed': {'date-parts': [[2024, 7, 2]], 'date-time': '2024-07-02T00:23:13Z', 'timestamp': 1719879793871}, 'posted': {'date-parts': [[2024]]}, 'group-title': 'SSRN', 'reference-count': 0, 'publisher': 'Elsevier BV', 'content-domain': {'domain': [], 'crossmark-restriction': False}, 'DOI': '10.2139/ssrn.4876852', 'type': 'posted-content', 'created': {'date-parts': [[2024, 7, 1]], 'date-time': '2024-07-01T10:14:50Z', 'timestamp': 1719828890000}, 'source': 'Crossref', 'is-referenced-by-count': 0, 'title': 'Proteomic Analysis Identifies the Glutathione Synthesizing Enzyme Gclc as an Andrographolide ' 'Target and a Protective Factor Against Sars-Cov-2 Infection', 'prefix': '10.2139', 'author': [ {'given': 'Jarinya', 'family': 'Chaopreecha', 'sequence': 'first', 'affiliation': []}, {'given': 'Nut', 'family': 'Phueakphud', 'sequence': 'additional', 'affiliation': []}, {'given': 'Ampa', 'family': 'Suksatu', 'sequence': 'additional', 'affiliation': []}, {'given': 'Sucheewin', 'family': 'Krobthong', 'sequence': 'additional', 'affiliation': []}, {'given': 'Suwimon', 'family': 'Manopwisedjaroen', 'sequence': 'additional', 'affiliation': []}, {'given': 'Nattawadee', 'family': 'Panyain', 'sequence': 'additional', 'affiliation': []}, {'given': 'Suradej', 'family': 'Hongeng', 'sequence': 'additional', 'affiliation': []}, {'given': 'Arunee', 'family': 'Thitithanyanont', 'sequence': 'additional', 'affiliation': []}, { 'given': 'Patompon', 'family': 'Wongtrakoongate', 'sequence': 'additional', 'affiliation': []}], 'member': '78', 'container-title': [], 'original-title': [], 'deposited': { 'date-parts': [[2024, 7, 1]], 'date-time': '2024-07-01T10:14:50Z', 'timestamp': 1719828890000}, 'score': 1, 'resource': {'primary': {'URL': 'https://www.ssrn.com/abstract=4876852'}}, 'subtitle': [], 'short-title': [], 'issued': {'date-parts': [[2024]]}, 'references-count': 0, 'URL': 'http://dx.doi.org/10.2139/ssrn.4876852', 'relation': {}, 'subject': [], 'published': {'date-parts': [[2024]]}, 'subtype': 'preprint'}

Download Image

Please send us corrections, updates, or comments. c19early involves the extraction of 100,000+ datapoints from thousands of papers. Community updates help ensure high accuracy. Treatments and other interventions are complementary. All practical, effective, and safe means should be used based on risk/benefit analysis. No treatment or intervention is 100% available and effective for all current and future variants. We do not provide medical advice. Before taking any medication, consult a qualified physician who can provide personalized advice and details of risks and benefits based on your medical history and situation. FLCCC and WCH provide treatment protocols.

Submit

Post

@CovidAnalysis

FAQ

Public domain CC0 1.0