{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,3,30]],"date-time":"2025-03-30T21:40:04Z","timestamp":1743370804262,"version":"3.40.3"},"reference-count":82,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,3,27]],"date-time":"2025-03-27T00:00:00Z","timestamp":1743033600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0"},{"start":{"date-parts":[[2025,3,27]],"date-time":"2025-03-27T00:00:00Z","timestamp":1743033600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0"}],"funder":[{"DOI":"10.13039\/501100004901","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de Minas Gerais","doi-asserted-by":"publisher","award":["APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18"],"award-info":[{"award-number":["APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18","APQ-00567-18"]}],"id":[{"id":"10.13039\/501100004901","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"DOI":"10.1038\/s41598-025-94283-9","type":"journal-article","created":{"date-parts":[[2025,3,27]],"date-time":"2025-03-27T23:16:08Z","timestamp":1743117368000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["In vitro enzymatic and cell culture assays for SARS-CoV-2 main protease interaction with ambenonium"],"prefix":"10.1038","volume":"15","author":[{"given":"Juliana \u00c2ngelo","family":"de Souza","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Isabela","family":"de Souza Gomes","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luciana","family":"de Souza Fernandes","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luis Adan Flores","family":"Andrade","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luciana \u00c2ngelo","family":"de Souza","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vin\u00edcius","family":"de Almeida Paiva","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sheila Cruz","family":"Araujo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Leonardo Henrique Franca","family":"de Lima","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Roberto Sousa","family":"Dias","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Raquel Cardoso","family":"de Melo-Minardi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fl\u00e1vio Guimar\u00e3es","family":"da Fonseca","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S\u00e9rgio Oliveira","family":"de Paula","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sabrina","family":"de Azevedo Silveira","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2025,3,27]]},"reference":[{"key":"94283_CR1","doi-asserted-by":"publisher","first-page":"264","DOI":"10.1016\/j.ijid.2020.01.009","volume":"91","author":"DS Hui","year":"2020","unstructured":"Hui, D. S. et al. The continuing 2019-nCoV epidemic threat of novel coronaviruses to global health \u2014 The latest 2019 novel coronavirus outbreak in Wuhan. China. Int. J. Infect. Dis. 91, 264\u2013266 (2020).","journal-title":"China. Int. J. Infect. Dis."},{"key":"94283_CR2","doi-asserted-by":"publisher","first-page":"141","DOI":"10.1038\/s41579-020-00459-7","volume":"19","author":"B Hu","year":"2021","unstructured":"Hu, B. et al. Characteristics of SARS-CoV-2 and COVID-19. Nat. Rev. Microbiol. 19, 141\u2013154 (2021).","journal-title":"Nat. Rev. Microbiol."},{"key":"94283_CR3","unstructured":"World Health Organization. Coronavirus Disease (COVID-19). https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/coronavirus-disease-%28covid-19%29 (2024)."},{"key":"94283_CR4","doi-asserted-by":"publisher","DOI":"10.1136\/bmjopen-2020-040729","volume":"10","author":"C Fumagalli","year":"2020","unstructured":"Fumagalli, C. et al. Clinical risk score to predict in-hospital mortality in COVID-19 patients: A retrospective cohort study. BMJ Open 10, e040729 (2020).","journal-title":"BMJ Open"},{"key":"94283_CR5","doi-asserted-by":"publisher","first-page":"181","DOI":"10.1038\/s41579-018-0118-9","volume":"17","author":"J Cui","year":"2019","unstructured":"Cui, J., Li, F. & Shi, Z. L. Origin and evolution of pathogenic coronaviruses. Nat. Rev. Microbiol. 17, 181\u2013192 (2019).","journal-title":"Nat. Rev. Microbiol."},{"key":"94283_CR6","doi-asserted-by":"publisher","first-page":"685","DOI":"10.1038\/s41579-021-00630-8","volume":"19","author":"H Yang","year":"2021","unstructured":"Yang, H. & Rao, Z. Structural biology of SARS-CoV-2 and implications for therapeutic development. Nat. Rev. Microbiol. 19, 685\u2013700 (2021).","journal-title":"Nat. Rev. Microbiol."},{"key":"94283_CR7","doi-asserted-by":"publisher","first-page":"199","DOI":"10.1007\/s15010-020-01516-2","volume":"49","author":"D Yesudhas","year":"2021","unstructured":"Yesudhas, D., Srivastava, A. & Gromiha, M. M. COVID-19 outbreak: History, mechanism, transmission, structural studies and therapeutics. Infection 49, 199\u2013213 (2021).","journal-title":"Infection"},{"key":"94283_CR8","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmcl.2020.127377","volume":"30","author":"S Ullrich","year":"2020","unstructured":"Ullrich, S. & Nitsche, C. The SARS-CoV-2 main protease as drug target. Bioorg. Med. Chem. Lett. 30, 127377 (2020).","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"94283_CR9","doi-asserted-by":"publisher","first-page":"481","DOI":"10.1055\/a-2095-0826","volume":"73","author":"S Kumar","year":"2023","unstructured":"Kumar, S. & Roy, V. Repurposing drugs: An empowering approach to drug discovery and development. Drug Res. 73, 481\u2013490 (2023).","journal-title":"Drug Res."},{"key":"94283_CR10","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3389\/fbinf.2023.1121591","volume":"3","author":"JP Santa-Maria","year":"2023","unstructured":"Santa-Maria, J. P., Wang, Y. & Camargo, L. M. Perspective on the challenges and opportunities of accelerating drug discovery with artificial intelligence. Front. Bioinform. 3, 1\u20135 (2023).","journal-title":"Front. Bioinform."},{"key":"94283_CR11","doi-asserted-by":"publisher","first-page":"967","DOI":"10.1016\/j.drudis.2021.11.023","volume":"27","author":"RSK Vijayan","year":"2022","unstructured":"Vijayan, R. S. K. et al. Enhancing preclinical drug discovery with artificial intelligence. Drug Discov. Today 27, 967\u2013984 (2022).","journal-title":"Drug Discov. Today"},{"key":"94283_CR12","doi-asserted-by":"publisher","first-page":"2076","DOI":"10.1016\/j.drudis.2019.06.014","volume":"24","author":"V Parvathaneni","year":"2019","unstructured":"Parvathaneni, V. et al. Drug repurposing: A promising tool to accelerate the drug discovery process. Drug Discov. Today 24, 2076\u20132085 (2019).","journal-title":"Drug Discov. Today"},{"key":"94283_CR13","doi-asserted-by":"publisher","first-page":"210","DOI":"10.1007\/s11684-020-0757-x","volume":"14","author":"L Ni","year":"2020","unstructured":"Ni, L. et al. Combination of western medicine and Chinese traditional patent medicine in treating a family case of COVID-19. Front. Med. 14, 210\u2013214 (2020).","journal-title":"Front. Med."},{"key":"94283_CR14","doi-asserted-by":"publisher","first-page":"241","DOI":"10.1016\/j.dsx.2020.03.011","volume":"14","author":"AK Singh","year":"2020","unstructured":"Singh, A. K. et al. Chloroquine and hydroxychloroquine in the treatment of COVID-19 with or without diabetes: A systematic search and a narrative review with a special reference to India and other developing countries. Diabet. Metab. Syndr. 14, 241\u2013246 (2020).","journal-title":"Diabet. Metab. Syndr."},{"key":"94283_CR15","doi-asserted-by":"publisher","first-page":"726","DOI":"10.1093\/bib\/bbaa288","volume":"22","author":"S Dotolo","year":"2021","unstructured":"Dotolo, S. et al. A review on drug repurposing applicable to COVID-19. Brief. Bioinform. 22, 726\u2013741 (2021).","journal-title":"Brief. Bioinform."},{"key":"94283_CR16","first-page":"1","volume":"9","author":"HM Menjist","year":"2021","unstructured":"Menjist, H. M., Dilnessa, T. & Jin, T. Structural basis of potential inhibitors targeting SARS-CoV-2 main protease. Front. Chem. 9, 1\u201311 (2021).","journal-title":"Front. Chem."},{"key":"94283_CR17","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3390\/pharmaceutics17010001","volume":"17","author":"SK Niazi","year":"2024","unstructured":"Niazi, S. K. & Mariam, Z. Computer-aided drug design and drug discovery: A prospective analysis. Pharmaceuticals 17, 1\u201322 (2024).","journal-title":"Pharmaceuticals"},{"key":"94283_CR18","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejps.2022.106324","volume":"181","author":"D Vemula","year":"2023","unstructured":"Vemula, D. et al. CADD, AI and ML in drug discovery: A comprehensive review. Eur. J. Pharm. Sci. 181, 106324 (2023).","journal-title":"Eur. J. Pharm. Sci."},{"key":"94283_CR19","volume":"17","author":"I de Souza Gomes","year":"2022","unstructured":"de Souza Gomes, I. et al. Computational prediction of potential inhibitors for SARS-CoV-2 main protease based on machine learning, docking, MM-PBSA calculations, and metadynamics. PLoS One 17, e0266729 (2022).","journal-title":"PLoS One"},{"key":"94283_CR20","doi-asserted-by":"publisher","first-page":"1349","DOI":"10.1007\/s40265-023-01937-x","volume":"83","author":"HA Blair","year":"2023","unstructured":"Blair, H. A. Correction to: Remdesivir: A review in COVID-19. Drugs 83, 1349 (2023).","journal-title":"Drugs"},{"key":"94283_CR21","doi-asserted-by":"publisher","first-page":"683","DOI":"10.7326\/M22-0281","volume":"175","author":"C Marzolini","year":"2022","unstructured":"Marzolini, C. et al. Prescribing nirmatrelvir-ritonavir: How to recognize and manage drug-drug interactions. Ann. Intern. Med. 175, 683\u2013690 (2022).","journal-title":"Ann. Intern. Med."},{"key":"94283_CR22","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.28732","volume":"95","author":"F Tian","year":"2023","unstructured":"Tian, F. et al. Nirmatrelvir\u2013ritonavir compared with other antiviral drugs for the treatment of COVID-19 patients: A systematic review and meta-analysis. J. Med. Virol. 95, e28454 (2023).","journal-title":"J. Med. Virol."},{"key":"94283_CR23","doi-asserted-by":"publisher","first-page":"1272565","DOI":"10.3389\/fcvm.2023.1256512","volume":"10","author":"KC Marques","year":"2023","unstructured":"Marques, K. C., Quaresma, J. A. S. & Falc\u00e3o, L. F. M. Cardiovascular autonomic dysfunction in \u201cLong COVID\u201d: pathophysiology, heart rate variability, and inflammatory markers. Front. Cardiovasc. Med. 10, 1272565 (2023).","journal-title":"Front. Cardiovasc. Med."},{"key":"94283_CR24","doi-asserted-by":"publisher","first-page":"39","DOI":"10.1186\/s40249-023-01092-1","volume":"12","author":"A Rezapour","year":"2023","unstructured":"Rezapour, A. Cost-effectiveness of remdesivir for the treatment of hospitalized patients with COVID-19: A systematic review. Infect. Dis. Poverty 12, 39 (2023).","journal-title":"Infect. Dis. Poverty"},{"key":"94283_CR25","doi-asserted-by":"publisher","DOI":"10.1016\/j.nmni.2020.100776","volume":"38","author":"C Prodromos","year":"2020","unstructured":"Prodromos, C. & Rumschlag, T. Hydroxychloroquine is effective, and consistently so when provided early, for COVID-19: A systematic review. New Microb. New Infect. 38, 100766 (2020).","journal-title":"New Microb. New Infect."},{"key":"94283_CR26","doi-asserted-by":"crossref","first-page":"1276898","DOI":"10.3389\/fpubh.2023.1276898","volume":"11","author":"O Martinuka","year":"2024","unstructured":"Martinuka, O. Methodological biases in observational hospital studies of COVID-19 treatment effectiveness: Pitfalls and potential. Front. Pub. Health 11, 1276898 (2024).","journal-title":"Front. Pub. Health"},{"key":"94283_CR27","doi-asserted-by":"publisher","first-page":"365","DOI":"10.1097\/EDE.0000000000001598","volume":"34","author":"A Breskin","year":"2023","unstructured":"Breskin, A. et al. Effectiveness of remdesivir treatment protocols among patients hospitalized with COVID-19: A target trial emulation. Epidemiology 34, 365\u2013375 (2023).","journal-title":"Epidemiology"},{"key":"94283_CR28","doi-asserted-by":"publisher","first-page":"19763","DOI":"10.1038\/s41598-021-86471-0","volume":"11","author":"YL Weng","year":"2021","unstructured":"Weng, Y. L. et al. Molecular dynamics and in silico mutagenesis on the reversible inhibitor-bound SARS-CoV-2 main protease complexes reveal the role of lateral pocket in enhancing the ligand affinity. Sci. Rep. 11, 19763 (2021).","journal-title":"Sci. Rep."},{"key":"94283_CR29","doi-asserted-by":"publisher","first-page":"156","DOI":"10.17161\/rrnmf.v4i3.19548","volume":"4","author":"L Remijn-Nelissen","year":"2023","unstructured":"Remijn-Nelissen, L. et al. MGFA International Conference Proceedings Symptomatic pharmacological treatment of myasthenia gravis. RRNMF Neuromuscul. J. 4, 156\u2013166 (2023).","journal-title":"RRNMF Neuromuscul. J."},{"key":"94283_CR30","doi-asserted-by":"publisher","first-page":"5421","DOI":"10.1046\/j.1432-1327.2000.01606.x","volume":"267","author":"J O\u2019Brien","year":"2000","unstructured":"O\u2019Brien, J. et al. Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur. J. Biochem. 267, 5421\u20135426 (2000).","journal-title":"Eur. J. Biochem."},{"key":"94283_CR31","doi-asserted-by":"publisher","DOI":"10.1590\/0074-02760200342","volume":"115","author":"DB Ara\u00fajo","year":"2020","unstructured":"Ara\u00fajo, D. B. et al. SARS-CoV-2 isolation from the first reported patients in Brazil and establishment of a coordinated task network. Mem. Inst. Oswaldo Cruz 115, e200199 (2020).","journal-title":"Mem. Inst. Oswaldo Cruz"},{"key":"94283_CR32","doi-asserted-by":"publisher","first-page":"2087","DOI":"10.1002\/jmv.25985","volume":"92","author":"F Musaraat","year":"2020","unstructured":"Musaraat, F. et al. The anti-HIV drug nelfinavir mesylate (Viracept) is a potent inhibitor of cell fusion caused by the SARS-CoV-2 spike (S) glycoprotein warranting further evaluation as an antiviral against COVID-19 infections. J. Med. Virol. 92, 2087\u20132095 (2020).","journal-title":"J. Med. Virol."},{"key":"94283_CR33","doi-asserted-by":"crossref","unstructured":"Trott, O. & Olson, A. J. AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J. Comput. Chem. 31, 455\u2013461 (2010).","DOI":"10.1002\/jcc.21334"},{"key":"94283_CR34","first-page":"251","volume":"10","author":"MT Nelson","year":"1996","unstructured":"Nelson, M. T. et al. NAMD: A parallel, object-oriented molecular dynamics program. Int. J. Supercomput. Appl. High. Perform. Comput. 10, 251\u2013268 (1996).","journal-title":"Int. J. Supercomput. Appl. High. Perform. Comput."},{"issue":"25","key":"94283_CR35","doi-asserted-by":"publisher","first-page":"2135","DOI":"10.1002\/jcc.23354","volume":"34","author":"J Huang","year":"2013","unstructured":"Huang, J. & MacKerell, A. D. Jr. \"CHARMM36 all-atom additive protein force field: Validation based on comparison to NMR data. J. comput. chem. 34(25), 2135\u20132145 (2013).","journal-title":"J. comput. chem."},{"issue":"12","key":"94283_CR36","doi-asserted-by":"publisher","first-page":"3155","DOI":"10.1021\/ci3003649","volume":"52","author":"V Kenno","year":"2012","unstructured":"Kenno, V. et al. Automation of the CHARMM general force field (CGenFF) II: Assignment of bonded parameters and partial atomic charges. J. Chem. Inform. model. 52(12), 3155\u20133168 (2012).","journal-title":"J. Chem. Inform. model."},{"key":"94283_CR37","doi-asserted-by":"publisher","first-page":"1093","DOI":"10.1007\/s00894-009-0454-9","volume":"15","author":"E Durham","year":"2009","unstructured":"Durham, E. et al. Solvent accessible surface area approximations for rapid and accurate protein structure prediction. J. mol. model. 15, 1093\u20131108 (2009).","journal-title":"J. mol. model."},{"key":"94283_CR38","doi-asserted-by":"publisher","first-page":"449","DOI":"10.1517\/17460441.2015.1032936","volume":"10","author":"S Genheden","year":"2015","unstructured":"Genheden, S. & Ryde, U. The MM\/PBSA and MM\/GBSA methods to estimate ligand-binding affinities. Exp. Op. Drug Discov. 10, 449\u2013461 (2015).","journal-title":"Exp. Op. Drug Discov."},{"key":"94283_CR39","doi-asserted-by":"publisher","first-page":"278","DOI":"10.1021\/cb200394t","volume":"7","author":"AC Anderson","year":"2013","unstructured":"Anderson, A. C. Winning the arms race by improving drug discovery against mutating targets. ACS Chem. Biol. 7, 278\u2013288 (2013).","journal-title":"ACS Chem. Biol."},{"key":"94283_CR40","first-page":"705","volume":"17","author":"GA Holdgate","year":"2018","unstructured":"Holdgate, G. A., Meek, T. D. & Grimley, R. L. Mechanistic enzymology in drug discovery: A fresh perspective. Nat. Rev. 17, 705\u2013718 (2018).","journal-title":"Nat. Rev."},{"key":"94283_CR41","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1038\/s41467-021-27889-y","volume":"13","author":"J Lee","year":"2022","unstructured":"Lee, J. et al. X-ray crystallographic characterization of the SARS-CoV-2 main protease polyprotein cleavage sites essential for viral processing and maturation. Nat. Commun. 13, 208 (2022).","journal-title":"Nat. Commun."},{"key":"94283_CR42","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2022.167706","volume":"434","author":"J Li","year":"2022","unstructured":"Li, J. et al. Structural basis of main proteases of coronavirus bound to drug candidate PF-07304814. J. Mol. Biol. 434, 102230 (2022).","journal-title":"J. Mol. Biol."},{"key":"94283_CR43","doi-asserted-by":"publisher","first-page":"409","DOI":"10.1126\/science.abb3405","volume":"358","author":"L Zhang","year":"2020","unstructured":"Zhang, L. et al. Crystal structure of SARS-CoV-2 main protease provides a basis for design of improved \u03b1-ketoamide inhibitors. Science 358, 409\u2013412 (2020).","journal-title":"Science"},{"key":"94283_CR44","unstructured":"PyMol. The PyMOL Molecular Graphics System, Version 3.0. Schr\u00f6dinger, LLC."},{"key":"94283_CR45","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41598-020-79357-0","volume":"10","author":"JC Ferreira","year":"2020","unstructured":"Ferreira, J. C. & Rabeh, W. M. Biochemical and biophysical characterization of the main protease, 3-chymotrypsin-like protease (3CLpro) from the novel coronavirus SARS-CoV-2. Sci. Rep. 10, 1\u201311 (2020).","journal-title":"Sci. Rep."},{"key":"94283_CR46","doi-asserted-by":"publisher","first-page":"12437","DOI":"10.1038\/s41598-020-69337-9","volume":"10","author":"R Yoshino","year":"2020","unstructured":"Yoshino, R., Yasuo, N. & Sekijima, M. Identification of key interactions between SARS-CoV-2 main protease and inhibitor drug candidates. Sci. Rep. 10, 12437 (2020).","journal-title":"Sci. Rep."},{"key":"94283_CR47","doi-asserted-by":"publisher","DOI":"10.1038\/s42003-022-03084-7","author":"NT Nashed","year":"2022","unstructured":"Nashed, N. T. et al. Modulation of the monomer-dimer equilibrium and catalytic activity of SARS-CoV-2 main protease by a transition-state analog inhibitor. Commun. Biol. https:\/\/doi.org\/10.1038\/s42003-022-03084-7 (2022).","journal-title":"Commun. Biol."},{"key":"94283_CR48","doi-asserted-by":"publisher","DOI":"10.1111\/ene.16229","author":"NE Gilhus","year":"2024","unstructured":"Gilhus, N. E. et al. Generalized myasthenia gravis with acetylcholine receptor antibodies: A guide for treatment. Eur. J. Neurol. https:\/\/doi.org\/10.1111\/ene.16229 (2024).","journal-title":"Eur. J. Neurol."},{"key":"94283_CR49","doi-asserted-by":"publisher","first-page":"1833","DOI":"10.1016\/j.ijbiomac.2018.07.030","volume":"118","author":"A Arsalan","year":"2015","unstructured":"Arsalan, A. & Younus, H. Enzymes and nanoparticles: Modulation of enzymatic activity via nanoparticles. Int. J. Biol. Macromol. 118, 1833\u20131847 (2015).","journal-title":"Int. J. Biol. Macromol."},{"key":"94283_CR50","doi-asserted-by":"publisher","first-page":"6086","DOI":"10.1111\/febs.16124","volume":"289","author":"B Srinivasan","year":"2021","unstructured":"Srinivasan, B. A guide to Michaelis-Menten equation: Steady state and beyond. FEBS J. 289, 6086\u20136098 (2021).","journal-title":"FEBS J."},{"key":"94283_CR51","doi-asserted-by":"publisher","first-page":"5682","DOI":"10.20964\/2020.06.09","volume":"15","author":"K Nirmala","year":"2020","unstructured":"Nirmala, K., Manimegalai, B. & Rajendran, L. Steady-state substrate and product concentrations for non-Michaelis-Menten kinetics in an amperometric biosensor- hyperbolic function and Pad\u00e9 approximants method. Int. J. Electrochem. Sci. 15, 5682\u20135697 (2020).","journal-title":"Int. J. Electrochem. Sci."},{"key":"94283_CR52","first-page":"102","volume":"28","author":"EF Pettersen","year":"2023","unstructured":"Pettersen, E. F. et al. High-throughput kinetic studies in drug discovery. Drug Discov. Today 28, 102\u2013109 (2023).","journal-title":"Drug Discov. Today"},{"key":"94283_CR53","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1016\/B978-0-12-152819-5.50020-8","volume":"19","author":"A Sols","year":"1981","unstructured":"Sols, A. Multimodulation of enzyme activity. Curr. Top. Cell Regul. 19, 77\u2013101 (1981).","journal-title":"Curr. Top. Cell Regul."},{"key":"94283_CR54","doi-asserted-by":"publisher","first-page":"1490","DOI":"10.1038\/s41564-022-01191-z","volume":"7","author":"M Giovanetti","year":"2022","unstructured":"Giovanetti, M. et al. Genomic epidemiology of the SARS-CoV-2 epidemic in Brazil. Nat. Microbiol. 7, 1490\u20131500 (2022).","journal-title":"Nat. Microbiol."},{"key":"94283_CR55","doi-asserted-by":"publisher","first-page":"446","DOI":"10.1002\/bmb.21235","volume":"47","author":"TP Silverstein","year":"2019","unstructured":"Silverstein, T. P. When both Km and Vmax are altered, is the enzyme inhibited or activated?. Biochem. Mol. Biol. Educ. 47, 446\u2013449 (2019).","journal-title":"Biochem. Mol. Biol. Educ."},{"key":"94283_CR56","doi-asserted-by":"publisher","first-page":"114247","DOI":"10.1016\/j.foodres.2024.114247","volume":"184","author":"W Chen","year":"2024","unstructured":"Chen, W. et al. The effect of konjac glucomannan on enzyme kinetics and fluorescence spectrometry of digestive enzymes: An in vitro research from the perspective of macromolecule crowding. Food Res. Int. 184, 114247 (2024).","journal-title":"Food Res. Int."},{"key":"94283_CR57","doi-asserted-by":"publisher","first-page":"2476","DOI":"10.1021\/acs.jchemed.9b00399","volume":"96","author":"TP Silverstein","year":"2019","unstructured":"Silverstein, T. P. & Slade, K. Effects of macromolecular crowding on biological systems. J. Chem. Educ. 96, 2476\u20132487 (2019).","journal-title":"J. Chem. Educ."},{"key":"94283_CR58","doi-asserted-by":"publisher","first-page":"e1011054","DOI":"10.1371\/journal.pcbi.1011054","volume":"19","author":"N Ostrowska","year":"2023","unstructured":"Ostrowska, N., Feig, M. & Trylska, J. Varying molecular interactions explain aspects of crowder-dependent enzyme function of a viral protease. PLoS Comput. Biol. 19, e1011054 (2023).","journal-title":"PLoS Comput. Biol."},{"key":"94283_CR59","first-page":"100956","volume":"26","author":"XE Wilcox","year":"2021","unstructured":"Wilcox, X. E., Chung, C. & Slade, K. M. Macromolecular crowding effects on the kinetics of opposing reactions catalyzed by alcohol dehydrogenase. Biochem. Biophys. Rep. 26, 100956 (2021).","journal-title":"Biochem. Biophys. Rep."},{"key":"94283_CR60","doi-asserted-by":"publisher","first-page":"196","DOI":"10.3390\/bios12040196","volume":"12","author":"D Lavogina","year":"2022","unstructured":"Lavogina, D. et al. Revisiting the resazurin-based sensing of cellular viability: Widening the application horizon. Biosensors 12, 196 (2022).","journal-title":"Biosensors"},{"key":"94283_CR61","doi-asserted-by":"publisher","first-page":"101981","DOI":"10.1016\/j.molmet.2024.101981","volume":"87","author":"AH Abir","year":"2024","unstructured":"Abir, A. H. et al. Metabolic profiling of single cells by exploiting NADH and FAD fluorescence via flow cytometry. Mol. Metab. 87, 101981 (2024).","journal-title":"Mol. Metab."},{"key":"94283_CR62","doi-asserted-by":"publisher","first-page":"62","DOI":"10.1007\/s11274-023-03871-2","volume":"40","author":"N Chandel","year":"2024","unstructured":"Chandel, N. et al. Indigo production goes green: A review on opportunities and challenges of fermentative production. World J. Microbiol. Biotechnol. 40, 62 (2024).","journal-title":"World J. Microbiol. Biotechnol."},{"key":"94283_CR63","doi-asserted-by":"publisher","first-page":"1610","DOI":"10.1364\/OSAC.423082","volume":"4","author":"TM Cannon","year":"2021","unstructured":"Cannon, T. M. et al. Characterization of NADH fluorescence properties under one-photon excitation with respect to temperature, pH, and binding to lactate dehydrogenase. OSA Contin. 4, 1610\u20131625 (2021).","journal-title":"OSA Contin."},{"key":"94283_CR64","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-024-53126-9","author":"TE Pylaev","year":"2024","unstructured":"Pylaev, T. E. et al. High-throughput cell optoporation system based on Au nanoparticle layers mediated by resonant irradiation for precise and controllable gene delivery. Sci. Rep. https:\/\/doi.org\/10.1038\/s41598-024-53126-9 (2024).","journal-title":"Sci. Rep."},{"key":"94283_CR65","unstructured":"Riss, T. et al. Cytotoxicity assays: In vitro methods to measure dead cells. In Assay Guidance Manual (2019)."},{"key":"94283_CR66","doi-asserted-by":"publisher","first-page":"156","DOI":"10.3390\/bios14040156","volume":"14","author":"J Petiti","year":"2024","unstructured":"Petiti, J., Revel, L. & Divieto, C. Standard operating procedure to optimize resazurin-based viability assays. Biosensors 14, 156 (2024).","journal-title":"Biosensors"},{"key":"94283_CR67","doi-asserted-by":"publisher","first-page":"12500","DOI":"10.1021\/acs.jmedchem.2c01005","volume":"65","author":"G La Monica","year":"2022","unstructured":"La Monica, G. et al. Targeting SARS-CoV-2 main protease for treatment of COVID-19: Covalent inhibitors structure\u2013activity relationship insights and evolution perspectives. J. Med. Chem. 65, 12500\u201312534 (2022).","journal-title":"J. Med. Chem."},{"key":"94283_CR68","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.59026","author":"D Lemoine","year":"2020","unstructured":"Lemoine, D. et al. Probing the ionotropic activity of glutamate GluD2 receptor in HEK cells with genetically-engineered photopharmacology. eLife https:\/\/doi.org\/10.7554\/eLife.59026 (2020).","journal-title":"eLife"},{"key":"94283_CR69","doi-asserted-by":"publisher","first-page":"348","DOI":"10.1002\/jca.21868","volume":"36","author":"HN Bennani","year":"2020","unstructured":"Bennani, H. N. Treatment of refractory myasthenia gravis by double-filtration plasmapheresis and rituximab: A case series of nine patients and literature review. J. Clin. Apher. 36, 348\u2013363 (2020).","journal-title":"J. Clin. Apher."},{"key":"94283_CR70","doi-asserted-by":"publisher","first-page":"928","DOI":"10.1128\/JVI.00695-14","volume":"88","author":"Z Hu","year":"2014","unstructured":"Hu, Z. & Kuritzkes, D. R. Altered viral fitness and drug susceptibility in HIV-1 carrying mutations that confer resistance to nonnucleoside reverse and integrase strand transfer inhibitors. J. Virol. 88, 928\u2013976 (2014).","journal-title":"J. Virol."},{"key":"94283_CR71","doi-asserted-by":"publisher","DOI":"10.3389\/fchem.2021.819165","author":"Z Lv","year":"2021","unstructured":"Lv, Z. et al. Targeting SARS-CoV-2 proteases for COVID-19 antiviral development. Front Chem https:\/\/doi.org\/10.3389\/fchem.2021.819165 (2021).","journal-title":"Front Chem"},{"key":"94283_CR72","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1093\/pcmedi\/pbab001","volume":"4","author":"M Hosseini","year":"2021","unstructured":"Hosseini, M. et al. Computational molecular docking and virtual screening revealed promising SARS-CoV-2 drugs. Precis. Clin. Med. 4, 1\u201316 (2021).","journal-title":"Precis. Clin. Med."},{"key":"94283_CR73","doi-asserted-by":"publisher","DOI":"10.1186\/s13578-021-00564-x","author":"Z Chen","year":"2021","unstructured":"Chen, Z. et al. Ginkgolic acid and anacardic acid are specific covalent inhibitors of SARS-CoV-2 cysteine proteases. Cell Biosci. https:\/\/doi.org\/10.1186\/s13578-021-00564-x (2021).","journal-title":"Cell Biosci."},{"key":"94283_CR74","doi-asserted-by":"publisher","first-page":"678","DOI":"10.1038\/s41422-020-0356-z","volume":"30","author":"C Ma","year":"2020","unstructured":"Ma, C. et al. Boceprevir, GC-376, and calpain inhibitors II, XII inhibit SARS-CoV-2 viral replication by targeting the viral main protease. Cell Res. 30, 678\u2013692 (2020).","journal-title":"Cell Res."},{"key":"94283_CR75","doi-asserted-by":"publisher","first-page":"7568","DOI":"10.1021\/jacs.2c00853","volume":"144","author":"J Zhou","year":"2022","unstructured":"Zhou, J. et al. Fast and effective prediction of the absolute binding free energies of covalent inhibitors of SARS-CoV-2 main protease and 20S proteasome. J. Am. Chem. Soc. 144, 7568\u20137572 (2022).","journal-title":"J. Am. Chem. Soc."},{"issue":"21","key":"94283_CR76","doi-asserted-by":"publisher","first-page":"14267","DOI":"10.1021\/acs.jmedchem.2c01147","volume":"65","author":"PW Kenny","year":"2022","unstructured":"Kenny, P. W. Hydrogen-bond donors in drug design. J. Med. Chem. 65(21), 14267\u201314275 (2022).","journal-title":"J. Med. Chem."},{"key":"94283_CR77","doi-asserted-by":"publisher","first-page":"e1477","DOI":"10.1002\/wcms.1477","volume":"10","author":"LJ Karas","year":"2020","unstructured":"Karas, L. J. et al. Hydrogen bond design principles. WIREs Comput. Mol. Sci. 10, e1477 (2020).","journal-title":"WIREs Comput. Mol. Sci."},{"key":"94283_CR78","unstructured":"PubChem. Compound summary: Ambenonium. https:\/\/pubchem.ncbi.nlm.nih.gov\/compound\/Ambenonium. Accessed (2024)."},{"key":"94283_CR79","doi-asserted-by":"publisher","first-page":"370","DOI":"10.1039\/D0MD00367K","volume":"12","author":"R Oerlemans","year":"2021","unstructured":"Oerlemans, R. et al. Repurposing the HCV NS3\u20134A protease drug boceprevir as COVID-19 therapeutics. R. Soc. Med. Chem. 12, 370\u2013379 (2021).","journal-title":"R. Soc. Med. Chem."},{"key":"94283_CR80","doi-asserted-by":"publisher","first-page":"167118","DOI":"10.1016\/j.jmb.2021.167118","volume":"433","author":"GD Noske","year":"2021","unstructured":"Noske, G. D. et al. A Crystallographic Snapshot of SARS-CoV-2 main protease maturation process. J. Mol. Biol. 433, 167118 (2021).","journal-title":"J. Mol. Biol."},{"key":"94283_CR81","doi-asserted-by":"publisher","first-page":"1435","DOI":"10.1038\/s42003-024-07138-w","volume":"7","author":"E Fornasier","year":"2024","unstructured":"Fornasier, E. et al. Allostery in homodimeric SARS-CoV-2 main protease. Commun. Biol. 7, 1435 (2024).","journal-title":"Commun. Biol."},{"issue":"10","key":"94283_CR82","doi-asserted-by":"publisher","first-page":"1233","DOI":"10.1080\/17460441.2021.1942836","volume":"16","author":"T Tuccinard","year":"2021","unstructured":"Tuccinard, T. What is the current value of MM\/PBSA and MM\/GBSA methods in drug discovery?. Expert Opin. Drug Discov. 16(10), 1233\u20131237 (2021).","journal-title":"Expert Opin. Drug Discov."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-94283-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-94283-9","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-94283-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,3,30]],"date-time":"2025-03-30T21:22:54Z","timestamp":1743369774000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-94283-9"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,27]]},"references-count":82,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["94283"],"URL":"https:\/\/doi.org\/10.1038\/s41598-025-94283-9","relation":{},"ISSN":["2045-2322"],"issn-type":[{"type":"electronic","value":"2045-2322"}],"subject":[],"published":{"date-parts":[[2025,3,27]]},"assertion":[{"value":"5 August 2024","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"12 March 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"27 March 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"10606"}}