Inhibition of SARS-COV-2 Receptor-Binding domain/ACE2 by Bazedoxifene

Inhibition of SARS-COV-2 Receptor-Binding domain/ACE2 by Bazedoxifene | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Inhibition of SARS-COV-2 Receptor-Binding domain/ACE2 by Bazedoxifene olaposi omotuyi, Olusina M. Olatunji, Oyekanmi Nash, Babatunji Oyinloye, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-1869175/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Coronavirus spike glycoprotein (GP)-Receptor Binding Domain (RBD) and its interaction with host receptor angiotensin converting enzyme 2 (ACE2) is one of the most structurally understood but therapeutically untapped aspect of COVID-19 pathogenesis. In this study, Fragment-based lead discovery was conducted to identify potential lead anti-SARS-COV-2 drug candidate acting at RBD/ACE2 interface. From the original 12000 chemical building blocks, benzimidazole (BAZ) scaffold was identified. Fingerprint-based similarity search of the FDA-approved drug library for BAZ-containing compounds identified 12 drugs with the benzimidazole-like substructure. When these compounds were re—docked into GP/ACE2 interface, the consensus docking identified bazedoxifene as the hit. In vitro RBD/ACE2 inhibition kinetics showed micromolar IC50 value (1.237 μM) in the presence of bazedoxifene. Molecular dynamics simulation of RBD/ACE2 in the presence BAZ resulted in loss of contact and specific hydrogen-bond interaction required for RBD/ACE2 stability. Taken together, these findings identified benzimidazole scaffold as a building block for developing novel RBD/ACE2 complex inhibitor and provided mechanistic basis for the use of bazedoxifene as a repurposable drug for the treatment of COVID-19 acting at RBD/ACE2 interface. COVID-19 Receptor binding domain ACE2 Bazedoxifene SARS-COV2 Figures Figure 1 Figure 2 Introduction Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease 2019 (COVID-19); a pneumonia-like illness responsible for the COVID-19 pandemic infecting approximately 25 million people and causing more than 835,000 deaths (Johns Hopkins, Coronavirus resource center) and counting. Two general approaches have been adopted for treating COVID-19 cases; vaccine and therapeutics development. Small molecules such as remdesivir, hydroxychloroquine, ribavirin, and ritonavir were initially repurposed as therapeutics for COVID-19 1 but have not performed satisfactorily clinically. New drugs specifically developed for COVID-19 by Merck (molnupiravir), Pfizer (ritonavir in combination with PF-07321332) 2 have been adjudged more efficacious 3 . Research efforts at developing plant-based therapeutic agents is also worthy of mention 4 . Although comparatively, vaccines have been more successful in combating COVID-19 than small molecular weight drugs within clinical settings, this success has been threatened by the fast-rising cases of mutations 5 . Thus, making discovery and development of novel anti-COVDI-19 agents highly desirable in the long run. To this end, 12,000 chemical building blocks were computationally screened to identify possible virus entry inhibitor candidates acting at the spike glycoprotein-angiotensin (RBD) converting enzyme II (ACE2) interface 6 . Whilst most of the previous successful efforts have targeted SARS-COV-2 proteases 7 , the current strategy provides a complementary approach such that a successful candidate will potentially reduce virus active tissue reservoir as the distribution of ACE2 has been confirmed in most cells with the exception of matured erythrocytes 8 . This study identified benzimidazole fragment as particularly interesting, and when compounds bearing benzimidazole scaffold were searched from FDA-approved library, we have identified bazedoxifene as the most likely candidate inhibitor of RBD/ACE2 interaction through molecular docking, atomistic simulation and validation by inhibitory kinetics studies. The previous reports that benzimidazole is a potent entry inhibitor to hepatitis C virus 9 somewhat improves our overall confidence that bazedoxifene may represent a repurposable anti-COVID-19 agents. Materials And Methods 2D Fingerprint search, Ligand Library, and Protein Preparation The database of FDA-approved drugs (open.fda.gov) were retrieved and filtered (Datawarrior Suite) using the molecular weight cutoff 200 ≥ 700 g/mol. The residual compounds were searched (open babel) for benzimidazole substructure using a path-based fingerprint (FP2). Each compound was scored using the Tanimoto coefficient and plotted as a function of the population size using Graphed prism. Unless otherwise stated, all ligands for molecular docking were incorporated into Maestro and prepared using the Schrodinger suite version 2018-1. Employing Ligprep 10 , Epik 11 with OPLS3 force field for protonation, stereoisomerization, tautomers generation, and to achieve biological conformers. Energy minimization was achieved for all tautomeric states at a pH of 7±2. 2019-nCoV RBD/ACE2 Complex (2.9 Å) was retrieved from PDB 12 with the PDB ID 6M17 13 . The complex was prepared using the protein preparation wizard module in maestro 11.5 by default settings. The complex was optimized and then minimized using the OPLS3 force field by converging heavy atoms to RMSD of 0.3Å. The complex interaction interface for docking was generated with the receptor grid generation tool in maestro 11.5. The interaction interface between RBD/ACE2 complex was defined by amino acid residues: K417/Q409/R403/D405/R408/Y505 (RBD) and D30/N33/T324/F356/(ACE2), with van der Waals scaling factor 1.00 and charge cutoff of 0.25 around the complex interaction interface residues. Molecular Docking Molecular docking was conducted using Autodock Vina 14 , and Glide module on maestro 11.5 15 . The library of compounds was docked into the interface of the target using the standard precision (SP) and Extra precision (XP) algorithm 16 , applying a scaling factor of 0.8 and partial charge cutoff of 0.15, the ligand was handled as flexible. Lastly, the binding affinity of the receptor-ligand complex was ranked according to Glide score (SP and XP) while Vina score was utilized for the Autodock vina program. Molecular Dynamics Simulation Biosystems setup for atomistic simulation The input files for MD simulation for each of the biosystems (apo-RBD/ACE2, BAZ-RBD/ACE2; ACE2=aa21-559; RBD=aa336-518) was generated using CHARMM-GUI webserver ( www.charmm-gui.org ). BAZ parametrization was performed using ParamChem service (https://cgenff.paramchem.org) as implemented on CHARMM-GUI webserver. During biosystem build-up, protein and ligand atoms were defined by CHARMM36 all-atom additive force field parameters, solvated in TIP3P explicit water model and neutralized with Na + /CL − at 0.15 M 17 . Details of biosystem setup using CHARMM-GUI have been previously published 18-20 . Molecular dynamics simulation was run on GROMACS ( ver. 5) 21 software. During equilibration, the biosystems were subjected to constant pressure and temperature (NPT; 310K, 1 bar) conditions using Berendsen temperature and pressure coupling algorithms as implemented in GROMACS. Van der Waals interactions were estimated at 10 Å, long-range electrostatic interactions were computed using particle mesh Ewald (PME) summation scheme while equation of atomic motion was integrated using the leap-frog algorithm at 2 fs time step for a total time of 50 ns with positional restraints imposed on the heavy atoms in all directions. Production simulations were performed at 100 ns with the removal of atomic restraints. All calculations were performed on Super-Micro workstations (32-E2600 Intel Xeon CPUs, M6000 GPUs Accelerator PCI-E x16 Card/node) housed at Bio-Computing Research Unit (B-cRU), Mols and Sims , Ado Ekiti, Ekiti State, Nigeria. Post-simulation trajectory analysis and MMPBSA calculation. Unless otherwise stated, 3D atomic representations were drawn using Visual Molecular Dynamics (VMD) software 22 . Interatomic distance was calculated using g_dist tool inbuilt in GROMACS software. Population plots from distance values were plotted using GraphPad-Prism software. Binding free energy was calculated GROMACS Tool for high-throughput MM-PBSA calculations ( g_mmpbsa ) algorithm as described 23 . All line graphs were plotted as mean of 2 independent runs using GraphPad prism (ver 6.0e, 2014). RBD/ACE2 complex inhibition kinetics Bazedoxifene (Cat: HY-A0031/CS-0932, Lot: 58414) was dissolved in water aliquoted at concentrations 0.01, 0.03, 0.1, 0.3, 1.0, 3.0 mM in triplicates. RBD/ACE-2 interaction inhibition kinetics study was performed using COVID-19-Spike-ACE-2 binding assay kit (RayBiotech, Inc, Cat #: CoV-SACE2-1) following the manufacturer’s instructions. Results Virtual screening identifies benzimidazole fragment as high-affinity binder at RBD/ACE2 interface. Fragment-based lead discovery (FBLD) represents an efficient approach to drug discovery, taking advantage of the highly diverse fragment libraries (12,000 compounds) available at the Chem Bridge Corporation ( https://www.chembridge.com ), a virtual screening performed to identify potential hits at the RBD/ACE2 interface. Whilst most of the fragments had docking scores between -1.0 and -6.0 kcal/mol, three compounds namely: 1-(5-Nitro-1H-benzimidazol-2-yl)ethanol, 1-(2-piperidin-1-ylethyl)-1H-pyrazol-4-amine and 5-(3-(aminoethyk)-1,2,4-oxadiazol-5-yl)pyrrolidine-2-one have docking scores of -7.0 kcal/mol (Fig. 1A). Although, -7.0 kcal/mol docking score is weak, instead of subsequent growing and/or combining the lead fragments to produce other leads with a higher affinity and improved physicochemical properties, we rather screened the FDA-library for chemical compounds containing benzimidazole fragment as 1-(5-Nitro-1H-benzimidazol-2-yl)ethanol had the best docking score overall. 2D fingerprint search was conducted into the database of FDA-approved drugs to identify and rank (Tanimoto coefficient) compounds with benzimidazole scaffold which can be rapidly repurposed as entry inhibitors in the current pandemic. The best ranked compounds were twelve (12) including albendazole(1), etonitazene(2), galeterone(3), lansoprazole(4), mavatrep(5), mebendazole(6), omeprazole(7), pantoprazole(8), rabeprazole(9), tenatoprazole(10), triclabendazole(11), and bazedoxifene(12) (Fig. 1B). When these compounds were docked into RBD/ACE2 interface using consensus scoring method 24 , bazedoxifene (BAZ) was identified as exhibiting the lowest binding score (-6.24 kcal/mol ) (Supplementary Table 1.0). An overview of the binding position within the RBD/ACE2 intercalates between a network of aromatic and charged amino acids at the RBD/ACE2 interface; some of the key residues proximal to BAZ binding pocket include: D30/N33/T324/F356/ (ACE2) and K417/Q409/R403/D405/R408/Y505 (RBD) (Fig. 1B, ii ). These amino acids have identified as key to successful initiation of cellular invasion 25 . When the finding was validated inSARS-COV-2 spike-ACE2 binding assay, a mean IC50 value of 1.237 mM was obtained (Fig. 1B, iii ) thus, strongly suggesting that BAZ is a micromolar level inhibitor of RBD/ACE2 interaction and a potential drug in the current pandemic. Molecular Dynamics Simulation identified the atomistic basis for RBD/ACE2 inhibition Molecular dynamics (MD) simulation has become a routine in silico tools for drug discovery due to its key advantage of incorporating structural flexibility and entropic effects into target-drug interaction, which is limited in molecular docking. Thus, MD simulation is more accurate when monitoring atomistic motions and kinetics associated with drug–target recognition and binding 26 . Since our group has previous experience in deploying MD simulation for delineating mechanism of drug-target interaction in complexes such as lysophosphatidate-LPA1 20 , nefiracetam-NMDA receptors 27 , we therefore deployed MD simulation for the current studies in order to provide better insight into the mechanism of inhibition. First, investigation was conducted into the center of mass distance between residues constituting the interface (Fig. 2A, i ) showed that at approximately 40 ns, the distance reduced to 0.4 nm in apo state, indicating contact establishment; the presence of BAZ resulted in loss of contact with mean distance 0.8~1.2 nm starting at 40 ns till the end of the simulation. (Fig. 2A, ii ). Three amino acid pairs were further studied providing insight into loss of contact. Glu23/Tyr473 (Fig. 2A, iii ) and Asp30/Lys471 (Fig. 2A, i v) distance evolution with time strongly suggest loss of contact around 40 ns but Tyr41/T500 had already contact post equilibration in the presence of BAZ (Fig. 2A, v ). Clearly, the presence of BAZ within the RBD/ACE2 interface promotes loss of contact and possibly dissociation as suggested by the in vitro data. Lastly, the details of the amino acid network required for BAZ binding and the energy of binding were worked out using MMPBSA method 23 . Several amino acids at the RBD/ACE2 interface participate in BAZ binding (Fig. 2B, i ) but in terms of contribution, Glu329/Asp355/Asp350/D546 (ACE2) and Asp405/Glu406 and Asp420 (ACE2) have the most contribution using the energy -14.0 kj/mol energy cutoff, while the binding energy was estimated at -409.178±21.114 kj/mol (Table 2) These data strongly suggest high affinity of RBD/ACE2 interface for BAZ. Discussion In addition to the challenges of developing and distributing safe and effective SARS-CoV-2 (COVID-19) vaccines 28 , evidence for the presence of variants of concern (VOC) now mounts 29 . These VOCs in most cases accumulate mutations around the epitopes; causing viral escape or highly diminished vaccine efficacy 30 . It is therefore not unimaginable that cheaper, more effective therapeutic options must be investigated. One of the key successes here is the Pfizer-developed PF-07321332 2 . PF-07321332 is an orally bioavailable SARS-CoV-2 main protease inhibitor with excellent off-target selectivity and in vivo safety profile 3 . In current study, Fragment-based lead discovery-aided fingerprint-based similarity search 31 of the FDA-approved chemical library identified benzimidazole scaffold-containing compounds as potential inhibitor of RBD/ACE2 interaction. Indeed, some previous reports have identified benzimidazole scaffold-containing compounds as potent anti-viral agents 32, 33 . For instance, linear (dialkylamino)alkyl- derivatized 2-[(benzotriazol-1/2-yl)methyl]benzimidazoles exhibited nanomolar EC(50) activity on respiratory syncytial virus (RSV) 34 . The FDA-approved drug with the best docking result in this study is bazedoxifene. Bazedoxifene is a selective estrogen receptor modulator indicated for endometriosis 35 . It has also been indicated for postmenopausal osteoporosis 36 . The anti-SARS-COV-2 activity of bazedoxifene has been mentioned in some prior reports. For instance, in a study, it was demonstrated that bazedoxifene inhibits IL-6 signaling at therapeutic doses, leading to blockage of cytokine storm, ARDS and mortality in severe COVID-19 patients 37 . In another report, anti-SARS-CoV-2 activity with IC50 value of 3.44 μM, and CC50 value of 14.97 had been reported 38 . Here, we provide a mechanistic insight that bazedoxifene inhibits RBD/ACE2 complex. In conclusion, we have provided further evidence in support of bazedoxifene as a repurposable drug for the treatment of COVID-19 acting at RBD/ACE2 interface. These findings are of immediate importance as vaccine production, distribution and efficacy present a challenge in the global flight against COVID-19. Declarations Authors’ contributions Authors OO, and SF and ON conceived and designed the study. Authors OMO and BO performed molecular docking and complied the manuscript. OS and AI performed the kinetic assay studies. All authors prepared the final manuscript, and proofreading Funding This research is supported by funds from the Nigerian Innovative Research Management Structures (NiReMaS) DAAD DIES ProGRANT, and Afe Babalola University Research Foundation. Competing interests There are no competing interests declared by the authors. References 1. Rehman, S. U.; Rehman, S. U.; Yoo, H. H., COVID-19 challenges and its therapeutics. Biomed Pharmacother 2021, 142 , 112015. 2. Owen, D. R.; Allerton, C. M. N.; Anderson, A. S.; Aschenbrenner, L.; Avery, M.; Berritt, S.; Boras, B.; Cardin, R. D.; Carlo, A.; Coffman, K. J.; Dantonio, A.; Di, L.; Eng, H.; Ferre, R.; Gajiwala, K. S.; Gibson, S. A.; Greasley, S. E.; Hurst, B. L.; Kadar, E. P.; Kalgutkar, A. S.; Lee, J. C.; Lee, J.; Liu, W.; Mason, S. W.; Noell, S.; Novak, J. J.; Obach, R. S.; Ogilvie, K.; Patel, N. C.; Pettersson, M.; Rai, D. K.; Reese, M. R.; Sammons, M. F.; Sathish, J. G.; Singh, R. S. P.; Steppan, C. M.; Stewart, A. E.; Tuttle, J. B.; Updyke, L.; Verhoest, P. R.; Wei, L.; Yang, Q.; Zhu, Y., An oral SARS-CoV-2 M(pro) inhibitor clinical candidate for the treatment of COVID-19. Science 2021, 374 (6575), 1586-1593. 3. Heskin, J.; Pallett, S. J. C.; Mughal, N.; Davies, G. W.; Moore, L. S. P.; Rayment, M.; Jones, R., Caution required with use of ritonavir-boosted PF-07321332 in COVID-19 management. Lancet 2022, 399 (10319), 21-22. 4. Omotuyi, I. O.; Nash, O.; Ajiboye, B. O.; Olumekun, V. O.; Oyinloye, B. E.; Osuntokun, O. T.; Olonisakin, A.; Ajayi, A. O.; Olusanya, O.; Akomolafe, F. S.; Adelakun, N., Aframomum melegueta secondary metabolites exhibit polypharmacology against SARS-CoV-2 drug targets: in vitro validation of furin inhibition. Phytother Res 2021, 35 (2), 908-919. 5. Aleem, A.; Akbar Samad, A. B.; Slenker, A. K., Emerging Variants of SARS-CoV-2 And Novel Therapeutics Against Coronavirus (COVID-19). In StatPearls , Treasure Island (FL), 2022. 6. Choudhary, S.; Malik, Y. S.; Tomar, S., Identification of SARS-CoV-2 Cell Entry Inhibitors by Drug Repurposing Using in silico Structure-Based Virtual Screening Approach. Front Immunol 2020, 11 , 1664. 7. Li, J.; Lin, C.; Zhou, X.; Zhong, F.; Zeng, P.; Yang, Y.; Zhang, Y.; Yu, B.; Fan, X.; McCormick, P. J.; Fu, R.; Fu, Y.; Jiang, H.; Zhang, J., Structural Basis of the Main Proteases of Coronavirus Bound to Drug Candidate PF-07321332. J Virol 2022, 96 (8), e0201321. 8. Harmer, D.; Gilbert, M.; Borman, R.; Clark, K. L., Quantitative mRNA expression profiling of ACE 2, a novel homologue of angiotensin converting enzyme. FEBS Lett 2002, 532 (1-2), 107-10. 9. Vausselin, T.; Seron, K.; Lavie, M.; Mesalam, A. A.; Lemasson, M.; Belouzard, S.; Feneant, L.; Danneels, A.; Rouille, Y.; Cocquerel, L.; Foquet, L.; Rosenberg, A. R.; Wychowski, C.; Meuleman, P.; Melnyk, P.; Dubuisson, J., Identification of a New Benzimidazole Derivative as an Antiviral against Hepatitis C Virus. J Virol 2016, 90 (19), 8422-34. 10. Release, S., Maestro, Schrödinger. LLC, New York, NY, 2018-1, 2018 . 11. Brooks, W. H.; Daniel, K. G.; Sung, S. S.; Guida, W. C., Computational validation of the importance of absolute stereochemistry in virtual screening. J Chem Inf Model 2008, 48 (3), 639-45. 12. Berman, H. M.; Westbrook, J.; Feng, Z.; Gilliland, G.; Bhat, T. N.; Weissig, H.; Shindyalov, I. N.; Bourne, P. E., The Protein Data Bank. Nucleic Acids Res 2000, 28 (1), 235-42. 13. Yan, R.; Zhang, Y.; Li, Y.; Xia, L.; Guo, Y.; Zhou, Q., Structural basis for the recognition of SARS-CoV-2 by full-length human ACE2. Science 2020, 367 (6485), 1444-1448. 14. 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 2010, 31 (2), 455-61. 15. Sastry, G. M.; Adzhigirey, M.; Day, T.; Annabhimoju, R.; Sherman, W., Protein and ligand preparation: parameters, protocols, and influence on virtual screening enrichments. J Comput Aided Mol Des 2013, 27 (3), 221-34. 16. Friesner, R. A.; Murphy, R. B.; Repasky, M. P.; Frye, L. L.; Greenwood, J. R.; Halgren, T. A.; Sanschagrin, P. C.; Mainz, D. T., Extra precision glide: docking and scoring incorporating a model of hydrophobic enclosure for protein-ligand complexes. J Med Chem 2006, 49 (21), 6177-96. 17. Huang, J.; MacKerell, A. D., Jr., CHARMM36 all-atom additive protein force field: validation based on comparison to NMR data. J Comput Chem 2013, 34 (25), 2135-45. 18. Omotuyi, I. O., Ebola virus envelope glycoprotein derived peptide in human Furin-bound state: computational studies. J Biomol Struct Dyn 2015, 33 (3), 461-70. 19. Omotuyi, O. I.; Nash, O.; Safronetz, D.; Ojo, A. A.; Ogunwa, T. H.; Adelakun, N. S., T-705-modified ssRNA in complex with Lassa virus nucleoprotein exhibits nucleotide splaying and increased water influx into the RNA-binding pocket. Chem Biol Drug Des 2019, 93 (4), 544-555. 20. Omotuyi, O. I.; Nagai, J.; Ueda, H., Lys39-Lysophosphatidate Carbonyl Oxygen Interaction Locks LPA1 N-terminal Cap to the Orthosteric Site and partners Arg124 During Receptor Activation. Sci Rep 2015, 5 , 13343. 21. Van Der Spoel, D.; Lindahl, E.; Hess, B.; Groenhof, G.; Mark, A. E.; Berendsen, H. J., GROMACS: fast, flexible, and free. J Comput Chem 2005, 26 (16), 1701-18. 22. Hildebrand, P. W.; Rose, A. S.; Tiemann, J. K. S., Bringing Molecular Dynamics Simulation Data into View. Trends Biochem Sci 2019, 44 (11), 902-913. 23. Kumari, R.; Kumar, R.; Open Source Drug Discovery, C.; Lynn, A., g_mmpbsa--a GROMACS tool for high-throughput MM-PBSA calculations. J Chem Inf Model 2014, 54 (7), 1951-62. 24. Ren, X.; Shi, Y. S.; Zhang, Y.; Liu, B.; Zhang, L. H.; Peng, Y. B.; Zeng, R., Novel Consensus Docking Strategy to Improve Ligand Pose Prediction. J Chem Inf Model 2018, 58 (8), 1662-1668. 25. Chen, Y.; Guo, Y.; Pan, Y.; Zhao, Z. J., Structure analysis of the receptor binding of 2019-nCoV. Biochem Biophys Res Commun 2020 . 26. De Vivo, M.; Masetti, M.; Bottegoni, G.; Cavalli, A., Role of Molecular Dynamics and Related Methods in Drug Discovery. J Med Chem 2016, 59 (9), 4035-61. 27. Omotuyi, O. I.; Ueda, H., Molecular dynamics study-based mechanism of nefiracetam-induced NMDA receptor potentiation. Comput Biol Chem 2015, 55 , 14-22. 28. Forman, R.; Shah, S.; Jeurissen, P.; Jit, M.; Mossialos, E., COVID-19 vaccine challenges: What have we learned so far and what remains to be done? Health Policy 2021, 125 (5), 553-567. 29. Omotuyi, O.; Olubiyi, O.; Nash, O.; Afolabi, E.; Oyinloye, B.; Fatumo, S.; Femi-Oyewo, M.; Bogoro, S., SARS-CoV-2 Omicron spike glycoprotein receptor binding domain exhibits super-binder ability with ACE2 but not convalescent monoclonal antibody. Comput Biol Med 2022, 142 , 105226. 30. Paul, D.; Pyne, N.; Paul, S., Mutation profile of SARS-CoV-2 spike protein and identification of potential multiple epitopes within spike protein for vaccine development against SARS-CoV-2. Virusdisease 2021, 32 (4), 703-726. 31. Bajusz, D.; Racz, A.; Heberger, K., Why is Tanimoto index an appropriate choice for fingerprint-based similarity calculations? J Cheminform 2015, 7 , 20. 32. Tonelli, M.; Simone, M.; Tasso, B.; Novelli, F.; Boido, V.; Sparatore, F.; Paglietti, G.; Pricl, S.; Giliberti, G.; Blois, S.; Ibba, C.; Sanna, G.; Loddo, R.; La Colla, P., Antiviral activity of benzimidazole derivatives. II. Antiviral activity of 2-phenylbenzimidazole derivatives. Bioorg Med Chem 2010, 18 (8), 2937-53. 33. Vitale, G.; Corona, P.; Loriga, M.; Carta, A.; Paglietti, G.; Giliberti, G.; Sanna, G.; Farci, P.; Marongiu, M. E.; La Colla, P., 5-acetyl-2-arylbenzimidazoles as antiviral agents. Part 4. Eur J Med Chem 2012, 53 , 83-97. 34. Tonelli, M.; Paglietti, G.; Boido, V.; Sparatore, F.; Marongiu, F.; Marongiu, E.; La Colla, P.; Loddo, R., Antiviral activity of benzimidazole derivatives. I. Antiviral activity of 1-substituted-2-[(benzotriazol-1/2-yl)methyl]benzimidazoles. Chem Biodivers 2008, 5 (11), 2386-401. 35. Kulak, J., Jr.; Fischer, C.; Komm, B.; Taylor, H. S., Treatment with bazedoxifene, a selective estrogen receptor modulator, causes regression of endometriosis in a mouse model. Endocrinology 2011, 152 (8), 3226-32. 36. Palacios, S., Bazedoxifene acetate for the management of postmenopausal osteoporosis. Drugs Today (Barc) 2011, 47 (3), 187-95. 37. Smetana, K., Jr.; Rosel, D.; BrAbek, J., Raloxifene and Bazedoxifene Could Be Promising Candidates for Preventing the COVID-19 Related Cytokine Storm, ARDS and Mortality. In Vivo 2020, 34 (5), 3027-3028. 38. Jeon, S.; Ko, M.; Lee, J.; Choi, I.; Byun, S. Y.; Park, S.; Shum, D.; Kim, S., Identification of Antiviral Drug Candidates against SARS-CoV-2 from FDA-Approved Drugs. Antimicrob Agents Chemother 2020, 64 (7). Tables Table 1.0: Compounds with high Tanimoto coefficient with benzimidazole Compounds Vina Dock Glide XP Glide SP Mean Score Albendazole ( 1 ) -5.20 -2.84 -4.27 -4.10 etonitazene ( 2 ) -6.00 -2.48 -4.78 -4.42 galeterone ( 3 ) -6.80 -2.52 -4.03 -4.45 lansoprazole ( 4 ) -6.30 -5.23 -5.83 -5.79 mavatrep ( 5 ) -8.00 -3.81 -5.55 -5.79 mebendazole ( 6 ) -6.20 -3.41 -3.85 -4.49 omeprazole ( 7 ) -5.40 -5.48 -5.27 -5.38 pantoprazole ( 8 ) -6.50 -3.94 -5.81 -5.42 rabeprazole ( 9 ) -5.70 -5.74 -5.05 -5.50 tenatoprazole ( 10 ) -6.00 -4.03 -3.91 -4.65 triclabendazole ( 11 ) -5.70 -4.18 -4.69 -4.86 Bazedoxifene* ( 12 ) -7.40 -5.13 -6.20 -6.24 Table 2.0: MMPBSA Energy Estimation of Bazedoxifene at RBD/ACE2 interface. Energy Terms (Kj/mol) Compound van der Waal energy Electrostatic energy Polar solvation energy SASA energy Binding energy BAZ -128.110 ± 20.285 -431.479 ± 18.550 167.395 ± 39.644 -16.984 ± 2.664 -409.178 ± 21.114 Supplementary Files graphicalAbstractBAZE.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-1869175","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":121894364,"identity":"b5c53d9e-80e7-4d86-88f8-cee58d880910","order_by":0,"name":"olaposi omotuyi","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABIUlEQVRIiWNgGAWjYDACdjApAeNKyIHZPAwMjA24tDAjtIAUSRgTq4UBpoUhsYGQFn5m5mcPPvyxYOAXO3z8MW+bRXr/7B7DD28YbGQ3HOA9/AKLFslmNnPDmW0SDJKz0xKbedskcmfcOWMsOYchzXjDAb40CyxaDA4zmEnzAh1jcDvHEKyl4UaOgTQPw+HEDQd4zAywamH/Js3zB6ElXf5GjvFvHob/eLTwmEnzsCG0JBjcyAGKMBwAaTF+gNUvPOUgv/CA/DJzzjkJw413jpVZzjFINp4JNA1riLG3bwOGWJ0cv3TygQ9vyurk5W43b77xpsJOtu94j/EH7OHMBiJ4wExGNriDGUBRxiaBRwsU/EGVYsZhyygYBaNgFIwsAADPV18NVaio+gAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0002-5473-745X","institution":"ABUAD: Afe Babalola University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"olaposi","middleName":"","lastName":"omotuyi","suffix":""},{"id":121894365,"identity":"542bc42b-ec14-4bdd-bc5d-b641ed98deb8","order_by":1,"name":"Olusina M. Olatunji","email":"","orcid":"","institution":"ABUAD: Afe Babalola University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Olusina","middleName":"M.","lastName":"Olatunji","suffix":""},{"id":121894366,"identity":"2be6868d-a458-4782-9931-9cae77e87beb","order_by":2,"name":"Oyekanmi Nash","email":"","orcid":"","institution":"National Biotechnology Development Agency","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Oyekanmi","middleName":"","lastName":"Nash","suffix":""},{"id":121894367,"identity":"a8181e8a-df3f-45a6-817e-ce79e71681cd","order_by":3,"name":"Babatunji Oyinloye","email":"","orcid":"","institution":"ABUAD: Afe Babalola University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Babatunji","middleName":"","lastName":"Oyinloye","suffix":""},{"id":121894368,"identity":"52312a82-f9bf-4d4b-aba1-c863d76c3e5f","order_by":4,"name":"Opeyemi Soremekun","email":"","orcid":"","institution":"MRC Laboratories The Gambia: Medical Research Council Unit The Gambia at the London School of Hygiene and Tropical Medicine","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Opeyemi","middleName":"","lastName":"Soremekun","suffix":""},{"id":121894369,"identity":"a822d6f8-72da-4173-9d1e-05df0d9a43e1","order_by":5,"name":"Ayodeji Ijagbuji","email":"","orcid":"","institution":"Adekunle Ajasin University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ayodeji","middleName":"","lastName":"Ijagbuji","suffix":""},{"id":121894370,"identity":"efa1f5cf-678f-404b-8e66-47e5634f7b07","order_by":6,"name":"Segun Fatumo","email":"","orcid":"","institution":"National Biotechnology Development Agency","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Segun","middleName":"","lastName":"Fatumo","suffix":""}],"badges":[],"createdAt":"2022-07-18 09:20:03","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-1869175/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-1869175/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":24006869,"identity":"13800ca1-4163-4541-8f85-fcd232d30ab7","added_by":"auto","created_at":"2022-07-18 21:11:38","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1563463,"visible":true,"origin":"","legend":"\u003cp\u003e(A) Population count of docking score distribution of building block into RBD/ACE-2 interface; the inset show the best 3 ranked compounds 2D structures. Figure 1B: (i) Population count of FDA-approved drugs with benzimidazole scaffold based on Tanimoto coefficient scores (inset shows 12 compounds with benzimidazole scaffold substructure). (ii) A snapshot of the placement of bazedoxifene (12) within the RBD/ACE2 interface and a detailed projection of the amino acids contributing to the binding interaction.\u0026nbsp;Figure 1B. (iii) Dose-response curve fitting of RBD/ACE2 binding in the presence of graded concentration of bazedoxifene (BAZ).\u0026nbsp;\u003c/p\u003e\u003cp\u003e\u003cbr\u003e\u003c/p\u003e","description":"","filename":"Fig1.png","url":"https://assets-eu.researchsquare.com/files/rs-1869175/v1/06830e777b5bd700a5ae7acf.png"},{"id":24006483,"identity":"581cc39d-5196-4db4-b32f-13f2bfe97df5","added_by":"auto","created_at":"2022-07-18 21:06:38","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":966594,"visible":true,"origin":"","legend":"\u003cp\u003e(i) 3D representation of selected hydrogen-bond interaction at RBD (blue) and ACE2 (green) interface proximal to bazedoxifene (yellow stick) binding site. (ii-v)Time-dependent distance plots of RBD/ACE2, and selected residue pairs (see Y-axes). (D). (i) Energy contribution of amino acids (ACE2=green points, RBD=blue points) to\u0026nbsp;bazedoxifene\u0026nbsp;binding based on MMPBSA energy calculations; a 3D projection of the most contributing amino acids based on 14.0 kj/mol energy cutoff.\u003c/p\u003e\u003cp\u003e\u003cbr\u003e\u003c/p\u003e","description":"","filename":"Fig2.png","url":"https://assets-eu.researchsquare.com/files/rs-1869175/v1/425f34db45da942543e28792.png"},{"id":24721318,"identity":"01e6551e-5857-4e35-90ec-b593ec67e89b","added_by":"auto","created_at":"2022-08-03 13:53:59","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1217194,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-1869175/v1/cc5c1536-f93f-4695-8d53-78385d66aaff.pdf"},{"id":24006484,"identity":"dd58f54a-72ec-460d-b35d-46499591c6b6","added_by":"auto","created_at":"2022-07-18 21:06:38","extension":"docx","order_by":8,"title":"","display":"","copyAsset":false,"role":"supplement","size":145022,"visible":true,"origin":"","legend":"","description":"","filename":"graphicalAbstractBAZE.docx","url":"https://assets-eu.researchsquare.com/files/rs-1869175/v1/1305a0fe61e02b703fda8ef0.docx"}],"financialInterests":"","formattedTitle":"Inhibition of SARS-COV-2 Receptor-Binding domain/ACE2 by Bazedoxifene","fulltext":[{"header":"Introduction","content":"\u003cp\u003eSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease 2019 (COVID-19); a pneumonia-like illness responsible for the COVID-19 pandemic infecting approximately 25 million people and causing more than 835,000 deaths (Johns Hopkins, Coronavirus resource center) and counting. Two general approaches have been adopted for treating COVID-19 cases; vaccine and therapeutics development.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSmall molecules such as remdesivir, hydroxychloroquine, ribavirin, and ritonavir were initially repurposed as therapeutics for COVID-19\u0026nbsp;\u003csup\u003e1\u003c/sup\u003e but have not performed satisfactorily clinically. New drugs specifically developed for COVID-19 by Merck (molnupiravir), Pfizer (ritonavir in combination with PF-07321332)\u003csup\u003e2\u003c/sup\u003e have been adjudged more efficacious\u0026nbsp;\u003csup\u003e3\u003c/sup\u003e. Research efforts at developing plant-based therapeutic agents is also worthy of mention\u0026nbsp;\u003csup\u003e4\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAlthough comparatively, vaccines have been more successful in combating COVID-19 than small molecular weight drugs within clinical settings, this success has been threatened by the fast-rising cases of mutations\u003csup\u003e5\u003c/sup\u003e. Thus, making discovery and development of novel anti-COVDI-19 agents highly desirable in the long run. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTo this end, 12,000 chemical building blocks were computationally screened to identify possible virus entry inhibitor candidates acting at the spike glycoprotein-angiotensin (RBD) converting enzyme II (ACE2) interface\u0026nbsp;\u003csup\u003e6\u003c/sup\u003e. \u0026nbsp;Whilst most of the previous successful efforts have targeted SARS-COV-2 proteases\u0026nbsp;\u003csup\u003e7\u003c/sup\u003e, \u0026nbsp;the current strategy provides a complementary approach such that a successful candidate will potentially reduce virus active tissue reservoir as the distribution of \u0026nbsp;ACE2 has been confirmed in most cells with the exception of matured erythrocytes\u0026nbsp;\u003csup\u003e8\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThis study identified benzimidazole fragment as particularly interesting, and when compounds bearing benzimidazole scaffold were searched from FDA-approved library, we have identified bazedoxifene as the most likely candidate inhibitor of RBD/ACE2 interaction through molecular docking, atomistic simulation and validation by inhibitory kinetics studies. The previous reports that benzimidazole is a potent entry inhibitor to hepatitis C virus\u0026nbsp;\u003csup\u003e9\u003c/sup\u003e somewhat improves our overall confidence that bazedoxifene may represent a repurposable anti-COVID-19 agents. \u0026nbsp;\u003c/p\u003e"},{"header":"Materials And Methods","content":"\u003cp\u003e\u003cstrong\u003e2D Fingerprint search, Ligand Library, and Protein Preparation\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe database of FDA-approved drugs (open.fda.gov) were retrieved and filtered (Datawarrior Suite) using the molecular weight cutoff 200 \u0026ge; 700 g/mol. The residual compounds were searched (open babel) for benzimidazole substructure using a path-based fingerprint (FP2). Each compound was scored using the Tanimoto coefficient and plotted as a function of the population size using Graphed prism.\u003c/p\u003e\n\u003cp\u003eUnless otherwise stated, all ligands for molecular docking were incorporated into Maestro and prepared using the Schrodinger suite version 2018-1. Employing Ligprep\u0026nbsp;\u003csup\u003e10\u003c/sup\u003e, Epik\u0026nbsp;\u003csup\u003e11\u003c/sup\u003e with OPLS3 force field for protonation, stereoisomerization, tautomers generation, and to achieve biological conformers. Energy minimization was achieved for all tautomeric states at a pH of 7\u0026plusmn;2.\u003c/p\u003e\n\u003cp\u003e2019-nCoV RBD/ACE2 Complex (2.9 \u0026Aring;) was retrieved from PDB\u0026nbsp;\u003csup\u003e12\u003c/sup\u003e with the PDB ID 6M17\u003csup\u003e13\u003c/sup\u003e. The complex was prepared using the protein preparation wizard module in maestro 11.5 by default settings. The complex was optimized and then minimized using the OPLS3 force field by converging heavy atoms to RMSD of 0.3\u0026Aring;. The complex interaction interface for docking was generated with the receptor grid generation tool in maestro 11.5. The interaction interface between RBD/ACE2 complex was defined by \u0026nbsp;amino acid residues:\u0026nbsp;K417/Q409/R403/D405/R408/Y505 (RBD)\u0026nbsp;and\u0026nbsp;D30/N33/T324/F356/(ACE2), with van der Waals scaling factor 1.00 and charge cutoff of 0.25 around the complex interaction interface residues.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMolecular Docking\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMolecular docking was conducted using Autodock Vina \u003csup\u003e14\u003c/sup\u003e, and Glide module on maestro 11.5 \u003csup\u003e15\u003c/sup\u003e. The library of compounds was docked into the interface of the target using the standard precision (SP) and Extra precision (XP) algorithm \u003csup\u003e16\u003c/sup\u003e, applying a scaling factor of 0.8 and partial charge cutoff of 0.15, the ligand was handled as flexible. Lastly, the binding affinity of the receptor-ligand complex was ranked according to Glide score (SP and XP) while Vina score was utilized for the Autodock vina program.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMolecular Dynamics Simulation\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBiosystems setup for atomistic simulation\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe input files for MD simulation for each of the biosystems (apo-RBD/ACE2, BAZ-RBD/ACE2; ACE2=aa21-559; RBD=aa336-518) was generated using CHARMM-GUI webserver (\u003ca href=\"http://www.charmm-gui.org/\"\u003ewww.charmm-gui.org\u003c/a\u003e). BAZ parametrization was performed using ParamChem service (https://cgenff.paramchem.org) as implemented on CHARMM-GUI webserver. During biosystem build-up, protein and ligand atoms were defined by CHARMM36 all-atom additive force field parameters, solvated in TIP3P explicit water model and neutralized with Na\u003csup\u003e+\u003c/sup\u003e/CL\u003csup\u003e\u0026minus;\u003c/sup\u003e at 0.15 M\u0026nbsp;\u003csup\u003e17\u003c/sup\u003e. Details of biosystem setup using CHARMM-GUI have been previously published\u0026nbsp;\u003csup\u003e18-20\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eMolecular dynamics simulation was run on GROMACS (\u003cem\u003ever.\u003c/em\u003e 5)\u0026nbsp;\u003csup\u003e21\u003c/sup\u003e software. During equilibration, the biosystems were subjected to constant pressure and temperature (NPT; 310K, 1 bar) conditions using Berendsen temperature and pressure coupling algorithms as implemented in GROMACS. Van der Waals interactions were estimated at 10\u0026thinsp;\u0026Aring;, long-range electrostatic interactions were computed using particle mesh Ewald (PME) summation scheme while equation of atomic motion was integrated using the leap-frog algorithm at 2\u0026thinsp;fs time step for a total time of 50\u0026thinsp;ns with positional restraints imposed on the heavy atoms in all directions. Production simulations were performed at 100\u0026thinsp;ns with the removal of atomic restraints. All calculations were performed on Super-Micro workstations (32-E2600 Intel Xeon CPUs, M6000 GPUs Accelerator PCI-E x16 Card/node) housed at Bio-Computing Research Unit (B-cRU), \u003cem\u003eMols and Sims\u003c/em\u003e, Ado Ekiti, Ekiti State, Nigeria.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePost-simulation trajectory analysis and MMPBSA calculation.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eUnless otherwise stated, 3D atomic representations were drawn using Visual Molecular Dynamics (VMD) software \u003csup\u003e22\u003c/sup\u003e. Interatomic distance was calculated using \u003cem\u003eg_dist\u003c/em\u003e tool inbuilt \u0026nbsp;in GROMACS software. Population plots from distance values were plotted using GraphPad-Prism software. Binding free energy was calculated GROMACS Tool for high-throughput MM-PBSA calculations (\u003cem\u003eg_mmpbsa\u003c/em\u003e) algorithm as described \u003csup\u003e23\u003c/sup\u003e. All line graphs were plotted as mean\u0026thinsp;of 2 independent runs using GraphPad prism (ver 6.0e, 2014). \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eRBD/ACE2 complex inhibition kinetics\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBazedoxifene (Cat: HY-A0031/CS-0932, Lot: 58414) was dissolved in water aliquoted at concentrations 0.01, 0.03, 0.1, 0.3, 1.0, 3.0 mM in triplicates. \u0026nbsp;RBD/ACE-2 interaction inhibition kinetics study was performed using COVID-19-Spike-ACE-2 binding assay kit (RayBiotech, Inc, Cat #: CoV-SACE2-1) following the manufacturer\u0026rsquo;s instructions. \u0026nbsp;\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eVirtual screening identifies benzimidazole fragment as high-affinity binder at RBD/ACE2 interface.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFragment-based lead discovery (FBLD) represents an efficient approach to drug discovery, taking advantage of the highly diverse fragment libraries (12,000 compounds) available at the Chem Bridge Corporation (\u003ca href=\"https://www.chembridge.com/\"\u003ehttps://www.chembridge.com\u003c/a\u003e), a virtual screening performed to identify potential hits at the RBD/ACE2 interface. Whilst most of the fragments had docking scores between -1.0 and -6.0 kcal/mol, three compounds namely: 1-(5-Nitro-1H-benzimidazol-2-yl)ethanol, 1-(2-piperidin-1-ylethyl)-1H-pyrazol-4-amine and 5-(3-(aminoethyk)-1,2,4-oxadiazol-5-yl)pyrrolidine-2-one have docking scores of -7.0 kcal/mol (Fig. 1A). Although, -7.0 kcal/mol docking score is weak, instead of subsequent growing and/or combining the lead fragments to produce other leads with a higher affinity and improved physicochemical properties, we rather screened the FDA-library for chemical compounds containing benzimidazole fragment as 1-(5-Nitro-1H-benzimidazol-2-yl)ethanol had the best docking score overall.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e2D fingerprint search was conducted into the database of FDA-approved drugs to identify and rank (Tanimoto coefficient) compounds with benzimidazole scaffold which can be rapidly repurposed as entry inhibitors in the current pandemic. The best ranked compounds were twelve (12) including albendazole(1), etonitazene(2), galeterone(3), lansoprazole(4), mavatrep(5), mebendazole(6), omeprazole(7), pantoprazole(8), rabeprazole(9), tenatoprazole(10), triclabendazole(11), and bazedoxifene(12) (Fig. 1B). When these compounds were docked into RBD/ACE2 interface using consensus scoring method\u003csup\u003e24\u003c/sup\u003e, bazedoxifene (BAZ) was identified as exhibiting the lowest binding score (-6.24 \u003cem\u003ekcal/mol\u003c/em\u003e) (Supplementary\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eTable 1.0). An overview of the binding position within the RBD/ACE2 intercalates between a network of aromatic and charged amino acids at the RBD/ACE2 interface; some of the key residues proximal to BAZ binding pocket include: D30/N33/T324/F356/ (ACE2) and K417/Q409/R403/D405/R408/Y505 (RBD) (Fig. 1B, \u003cem\u003eii\u003c/em\u003e). These amino acids have identified as key to successful initiation of cellular invasion \u003csup\u003e25\u003c/sup\u003e. \u0026nbsp;When the finding was validated inSARS-COV-2 spike-ACE2 binding assay, a mean IC50 value of 1.237 \u0026nbsp;mM was obtained (Fig. 1B, \u003cem\u003eiii\u003c/em\u003e) thus, strongly suggesting that BAZ is a micromolar level inhibitor of RBD/ACE2 interaction and a potential drug \u0026nbsp;in the current pandemic.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMolecular Dynamics Simulation identified the atomistic basis for RBD/ACE2 inhibition\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMolecular dynamics (MD) simulation has become a routine in silico tools for drug discovery due to its key advantage of incorporating structural flexibility and entropic effects into target-drug interaction, which is limited in molecular docking. Thus, MD simulation is more accurate when monitoring atomistic motions and kinetics associated with drug\u0026ndash;target recognition and binding \u003csup\u003e26\u003c/sup\u003e. Since our group has previous experience in deploying MD simulation for delineating mechanism of drug-target interaction in complexes such as lysophosphatidate-LPA1 \u003csup\u003e20\u003c/sup\u003e, nefiracetam-NMDA receptors \u003csup\u003e27\u003c/sup\u003e, we therefore deployed MD simulation for the current studies in order to provide better insight into the mechanism of inhibition.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFirst, investigation was conducted into the center of mass distance between residues constituting the interface (Fig. 2A, \u003cem\u003ei\u003c/em\u003e) showed that at approximately 40 ns, the distance reduced to 0.4 nm in apo state, indicating contact establishment; the presence of BAZ resulted in loss of contact with mean distance 0.8~1.2 nm starting at 40 ns till the end of the simulation. (Fig. 2A, \u003cem\u003eii\u003c/em\u003e). \u0026nbsp;Three amino acid pairs were further studied providing insight into loss of contact. Glu23/Tyr473 (Fig. 2A, \u003cem\u003eiii\u003c/em\u003e) and Asp30/Lys471 (Fig. 2A, \u003cem\u003ei\u003c/em\u003ev) distance evolution with time strongly suggest loss of contact around 40 ns but Tyr41/T500 had already contact post equilibration in the presence of BAZ (Fig. 2A, \u003cem\u003ev\u003c/em\u003e). Clearly, the presence of BAZ within the RBD/ACE2 interface promotes loss of contact and possibly dissociation as suggested by the \u003cem\u003ein vitro\u003c/em\u003e data. \u0026nbsp; Lastly, the details of the amino acid network required for BAZ binding and the energy of binding were worked out using MMPBSA method \u003csup\u003e23\u003c/sup\u003e. \u0026nbsp;Several amino acids at the RBD/ACE2 interface participate in BAZ binding (Fig. 2B, \u003cem\u003ei\u003c/em\u003e) but in terms of contribution, Glu329/Asp355/Asp350/D546 (ACE2) and Asp405/Glu406 and Asp420 (ACE2) have the most contribution using the energy -14.0 \u003cem\u003ekj/mol\u003c/em\u003e energy cutoff, while the binding energy was estimated at -409.178\u0026plusmn;21.114 \u003cem\u003ekj/mol\u003c/em\u003e (Table 2) These data strongly suggest high affinity of RBD/ACE2 interface for BAZ.\u0026nbsp;\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn addition to the challenges of developing and distributing safe and effective SARS-CoV-2 (COVID-19) vaccines\u0026nbsp;\u003csup\u003e28\u003c/sup\u003e, evidence for the presence of variants of concern (VOC) now mounts\u003csup\u003e29\u003c/sup\u003e. These VOCs in most cases accumulate mutations around the epitopes; causing viral escape or highly diminished vaccine efficacy\u003csup\u003e30\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eIt is therefore not unimaginable that cheaper, more effective therapeutic options must be investigated. One of the key successes here is the Pfizer-developed PF-07321332\u0026nbsp;\u003csup\u003e2\u003c/sup\u003e. PF-07321332 is an orally bioavailable SARS-CoV-2 main protease inhibitor with excellent off-target selectivity and \u003cem\u003ein vivo\u003c/em\u003e safety profile\u0026nbsp;\u003csup\u003e3\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn current study, Fragment-based lead discovery-aided fingerprint-based similarity search\u0026nbsp;\u003csup\u003e31\u003c/sup\u003e of the FDA-approved chemical library identified benzimidazole scaffold-containing compounds as potential inhibitor of RBD/ACE2 interaction. Indeed, some previous reports have identified benzimidazole scaffold-containing compounds as potent anti-viral agents\u0026nbsp;\u003csup\u003e32, 33\u003c/sup\u003e. For instance, linear (dialkylamino)alkyl- derivatized 2-[(benzotriazol-1/2-yl)methyl]benzimidazoles exhibited nanomolar EC(50) \u0026nbsp;activity on respiratory syncytial virus (RSV)\u0026nbsp;\u003csup\u003e34\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe FDA-approved drug with the best docking result in this study is bazedoxifene. Bazedoxifene is a selective estrogen receptor modulator indicated for endometriosis\u0026nbsp;\u003csup\u003e35\u003c/sup\u003e. It has also been \u0026nbsp;indicated for postmenopausal osteoporosis\u0026nbsp;\u003csup\u003e36\u003c/sup\u003e. \u0026nbsp;The anti-SARS-COV-2 activity of bazedoxifene has been mentioned in some prior reports. For instance, in a study, it was demonstrated that bazedoxifene inhibits IL-6 signaling at therapeutic doses, leading to blockage of cytokine storm, ARDS and mortality in severe COVID-19 patients\u0026nbsp;\u003csup\u003e37\u003c/sup\u003e. In another report, anti-SARS-CoV-2 activity with IC50 value of 3.44\u0026thinsp;\u0026mu;M, and CC50 value of 14.97 \u0026nbsp;had been reported\u0026nbsp;\u003csup\u003e38\u003c/sup\u003e. \u0026nbsp; Here, we provide a mechanistic insight that bazedoxifene inhibits RBD/ACE2 complex.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn conclusion, we have provided further evidence in support of bazedoxifene as a repurposable drug for the treatment of COVID-19 acting at RBD/ACE2 interface. \u0026nbsp;These findings are of immediate importance as vaccine production, distribution and efficacy present a challenge in the global flight against COVID-19.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAuthors OO, and SF and ON conceived and designed the study. Authors OMO and BO performed molecular docking and complied the manuscript. OS and AI performed the kinetic assay studies. All authors prepared the final manuscript, and proofreading\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research is supported by funds from the Nigerian Innovative Research Management Structures (NiReMaS) DAAD DIES ProGRANT, and Afe Babalola University Research Foundation.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThere are no competing interests declared by the authors.\u003c/p\u003e"},{"header":"References","content":"\u003cp\u003e1.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Rehman, S. U.; \u0026nbsp;Rehman, S. U.; Yoo, H. H., COVID-19 challenges and its therapeutics. \u003cem\u003eBiomed Pharmacother\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2021,\u003c/strong\u003e \u003cem\u003e142\u003c/em\u003e, 112015.\u003c/p\u003e\n\u003cp\u003e2.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Owen, D. R.; \u0026nbsp;Allerton, C. M. N.; \u0026nbsp;Anderson, A. S.; \u0026nbsp;Aschenbrenner, L.; \u0026nbsp;Avery, M.; \u0026nbsp;Berritt, S.; \u0026nbsp;Boras, B.; \u0026nbsp;Cardin, R. D.; \u0026nbsp;Carlo, A.; \u0026nbsp;Coffman, K. J.; \u0026nbsp;Dantonio, A.; \u0026nbsp;Di, L.; \u0026nbsp;Eng, H.; \u0026nbsp;Ferre, R.; \u0026nbsp;Gajiwala, K. S.; \u0026nbsp;Gibson, S. A.; \u0026nbsp;Greasley, S. E.; \u0026nbsp;Hurst, B. L.; \u0026nbsp;Kadar, E. P.; \u0026nbsp;Kalgutkar, A. S.; \u0026nbsp;Lee, J. C.; \u0026nbsp;Lee, J.; \u0026nbsp;Liu, W.; \u0026nbsp;Mason, S. W.; \u0026nbsp;Noell, S.; \u0026nbsp;Novak, J. J.; \u0026nbsp;Obach, R. S.; \u0026nbsp;Ogilvie, K.; \u0026nbsp;Patel, N. C.; \u0026nbsp;Pettersson, M.; \u0026nbsp;Rai, D. K.; \u0026nbsp;Reese, M. R.; \u0026nbsp;Sammons, M. F.; \u0026nbsp;Sathish, J. G.; \u0026nbsp;Singh, R. S. P.; \u0026nbsp;Steppan, C. M.; \u0026nbsp;Stewart, A. E.; \u0026nbsp;Tuttle, J. B.; \u0026nbsp;Updyke, L.; \u0026nbsp;Verhoest, P. R.; \u0026nbsp;Wei, L.; \u0026nbsp;Yang, Q.; Zhu, Y., An oral SARS-CoV-2 M(pro) inhibitor clinical candidate for the treatment of COVID-19. \u003cem\u003eScience\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2021,\u003c/strong\u003e \u003cem\u003e374\u003c/em\u003e (6575), 1586-1593.\u003c/p\u003e\n\u003cp\u003e3.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Heskin, J.; \u0026nbsp;Pallett, S. J. C.; \u0026nbsp;Mughal, N.; \u0026nbsp;Davies, G. W.; \u0026nbsp;Moore, L. S. P.; \u0026nbsp;Rayment, M.; Jones, R., Caution required with use of ritonavir-boosted PF-07321332 in COVID-19 management. \u003cem\u003eLancet\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2022,\u003c/strong\u003e \u003cem\u003e399\u003c/em\u003e (10319), 21-22.\u003c/p\u003e\n\u003cp\u003e4.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Omotuyi, I. O.; \u0026nbsp;Nash, O.; \u0026nbsp;Ajiboye, B. O.; \u0026nbsp;Olumekun, V. O.; \u0026nbsp;Oyinloye, B. E.; \u0026nbsp;Osuntokun, O. T.; \u0026nbsp;Olonisakin, A.; \u0026nbsp;Ajayi, A. O.; \u0026nbsp;Olusanya, O.; \u0026nbsp;Akomolafe, F. S.; Adelakun, N., Aframomum melegueta secondary metabolites exhibit polypharmacology against SARS-CoV-2 drug targets: in vitro validation of furin inhibition. \u003cem\u003ePhytother Res\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2021,\u003c/strong\u003e \u003cem\u003e35\u003c/em\u003e (2), 908-919.\u003c/p\u003e\n\u003cp\u003e5.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Aleem, A.; \u0026nbsp;Akbar Samad, A. B.; Slenker, A. K., Emerging Variants of SARS-CoV-2 And Novel Therapeutics Against Coronavirus (COVID-19). In \u003cem\u003eStatPearls\u003c/em\u003e, Treasure Island (FL), 2022.\u003c/p\u003e\n\u003cp\u003e6.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Choudhary, S.; \u0026nbsp;Malik, Y. S.; Tomar, S., Identification of SARS-CoV-2 Cell Entry Inhibitors by Drug Repurposing Using in silico Structure-Based Virtual Screening Approach. \u003cem\u003eFront Immunol\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2020,\u003c/strong\u003e \u003cem\u003e11\u003c/em\u003e, 1664.\u003c/p\u003e\n\u003cp\u003e7.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Li, J.; \u0026nbsp;Lin, C.; \u0026nbsp;Zhou, X.; \u0026nbsp;Zhong, F.; \u0026nbsp;Zeng, P.; \u0026nbsp;Yang, Y.; \u0026nbsp;Zhang, Y.; \u0026nbsp;Yu, B.; \u0026nbsp;Fan, X.; \u0026nbsp;McCormick, P. J.; \u0026nbsp;Fu, R.; \u0026nbsp;Fu, Y.; \u0026nbsp;Jiang, H.; Zhang, J., Structural Basis of the Main Proteases of Coronavirus Bound to Drug Candidate PF-07321332. \u003cem\u003eJ Virol\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2022,\u003c/strong\u003e \u003cem\u003e96\u003c/em\u003e (8), e0201321.\u003c/p\u003e\n\u003cp\u003e8.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Harmer, D.; \u0026nbsp;Gilbert, M.; \u0026nbsp;Borman, R.; Clark, K. L., Quantitative mRNA expression profiling of ACE 2, a novel homologue of angiotensin converting enzyme. \u003cem\u003eFEBS Lett\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2002,\u003c/strong\u003e \u003cem\u003e532\u003c/em\u003e (1-2), 107-10.\u003c/p\u003e\n\u003cp\u003e9.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Vausselin, T.; \u0026nbsp;Seron, K.; \u0026nbsp;Lavie, M.; \u0026nbsp;Mesalam, A. A.; \u0026nbsp;Lemasson, M.; \u0026nbsp;Belouzard, S.; \u0026nbsp;Feneant, L.; \u0026nbsp;Danneels, A.; \u0026nbsp;Rouille, Y.; \u0026nbsp;Cocquerel, L.; \u0026nbsp;Foquet, L.; \u0026nbsp;Rosenberg, A. R.; \u0026nbsp;Wychowski, C.; \u0026nbsp;Meuleman, P.; \u0026nbsp;Melnyk, P.; Dubuisson, J., Identification of a New Benzimidazole Derivative as an Antiviral against Hepatitis C Virus. \u003cem\u003eJ Virol\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2016,\u003c/strong\u003e \u003cem\u003e90\u003c/em\u003e (19), 8422-34.\u003c/p\u003e\n\u003cp\u003e10.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Release, S., Maestro, Schr\u0026ouml;dinger. \u003cem\u003eLLC, New York, NY,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2018-1,\u003c/strong\u003e \u003cem\u003e2018\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003e11.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Brooks, W. H.; \u0026nbsp;Daniel, K. G.; \u0026nbsp;Sung, S. S.; Guida, W. C., Computational validation of the importance of absolute stereochemistry in virtual screening. \u003cem\u003eJ Chem Inf Model\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2008,\u003c/strong\u003e \u003cem\u003e48\u003c/em\u003e (3), 639-45.\u003c/p\u003e\n\u003cp\u003e12.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Berman, H. M.; \u0026nbsp;Westbrook, J.; \u0026nbsp;Feng, Z.; \u0026nbsp;Gilliland, G.; \u0026nbsp;Bhat, T. N.; \u0026nbsp;Weissig, H.; \u0026nbsp;Shindyalov, I. N.; Bourne, P. E., The Protein Data Bank. \u003cem\u003eNucleic Acids Res\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2000,\u003c/strong\u003e \u003cem\u003e28\u003c/em\u003e (1), 235-42.\u003c/p\u003e\n\u003cp\u003e13.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Yan, R.; \u0026nbsp;Zhang, Y.; \u0026nbsp;Li, Y.; \u0026nbsp;Xia, L.; \u0026nbsp;Guo, Y.; Zhou, Q., Structural basis for the recognition of SARS-CoV-2 by full-length human ACE2. \u003cem\u003eScience\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2020,\u003c/strong\u003e \u003cem\u003e367\u003c/em\u003e (6485), 1444-1448.\u003c/p\u003e\n\u003cp\u003e14.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Trott, O.; Olson, A. J., AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. \u003cem\u003eJ Comput Chem\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2010,\u003c/strong\u003e \u003cem\u003e31\u003c/em\u003e (2), 455-61.\u003c/p\u003e\n\u003cp\u003e15.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Sastry, G. M.; \u0026nbsp;Adzhigirey, M.; \u0026nbsp;Day, T.; \u0026nbsp;Annabhimoju, R.; Sherman, W., Protein and ligand preparation: parameters, protocols, and influence on virtual screening enrichments. \u003cem\u003eJ Comput Aided Mol Des\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2013,\u003c/strong\u003e \u003cem\u003e27\u003c/em\u003e (3), 221-34.\u003c/p\u003e\n\u003cp\u003e16.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Friesner, R. A.; \u0026nbsp;Murphy, R. B.; \u0026nbsp;Repasky, M. P.; \u0026nbsp;Frye, L. L.; \u0026nbsp;Greenwood, J. R.; \u0026nbsp;Halgren, T. A.; \u0026nbsp;Sanschagrin, P. C.; Mainz, D. T., Extra precision glide: docking and scoring incorporating a model of hydrophobic enclosure for protein-ligand complexes. \u003cem\u003eJ Med Chem\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2006,\u003c/strong\u003e \u003cem\u003e49\u003c/em\u003e (21), 6177-96.\u003c/p\u003e\n\u003cp\u003e17.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Huang, J.; MacKerell, A. D., Jr., CHARMM36 all-atom additive protein force field: validation based on comparison to NMR data. \u003cem\u003eJ Comput Chem\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2013,\u003c/strong\u003e \u003cem\u003e34\u003c/em\u003e (25), 2135-45.\u003c/p\u003e\n\u003cp\u003e18.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Omotuyi, I. O., Ebola virus envelope glycoprotein derived peptide in human Furin-bound state: computational studies. \u003cem\u003eJ Biomol Struct Dyn\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2015,\u003c/strong\u003e \u003cem\u003e33\u003c/em\u003e (3), 461-70.\u003c/p\u003e\n\u003cp\u003e19.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Omotuyi, O. I.; \u0026nbsp;Nash, O.; \u0026nbsp;Safronetz, D.; \u0026nbsp;Ojo, A. A.; \u0026nbsp;Ogunwa, T. H.; Adelakun, N. S., T-705-modified ssRNA in complex with Lassa virus nucleoprotein exhibits nucleotide splaying and increased water influx into the RNA-binding pocket. \u003cem\u003eChem Biol Drug Des\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2019,\u003c/strong\u003e \u003cem\u003e93\u003c/em\u003e (4), 544-555.\u003c/p\u003e\n\u003cp\u003e20.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Omotuyi, O. I.; \u0026nbsp;Nagai, J.; Ueda, H., Lys39-Lysophosphatidate Carbonyl Oxygen Interaction Locks LPA1 N-terminal Cap to the Orthosteric Site and partners Arg124 During Receptor Activation. \u003cem\u003eSci Rep\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2015,\u003c/strong\u003e \u003cem\u003e5\u003c/em\u003e, 13343.\u003c/p\u003e\n\u003cp\u003e21.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Van Der Spoel, D.; \u0026nbsp;Lindahl, E.; \u0026nbsp;Hess, B.; \u0026nbsp;Groenhof, G.; \u0026nbsp;Mark, A. E.; Berendsen, H. J., GROMACS: fast, flexible, and free. \u003cem\u003eJ Comput Chem\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2005,\u003c/strong\u003e \u003cem\u003e26\u003c/em\u003e (16), 1701-18.\u003c/p\u003e\n\u003cp\u003e22.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Hildebrand, P. W.; \u0026nbsp;Rose, A. S.; Tiemann, J. K. S., Bringing Molecular Dynamics Simulation Data into View. \u003cem\u003eTrends Biochem Sci\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2019,\u003c/strong\u003e \u003cem\u003e44\u003c/em\u003e (11), 902-913.\u003c/p\u003e\n\u003cp\u003e23.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Kumari, R.; \u0026nbsp;Kumar, R.; \u0026nbsp;Open Source Drug Discovery, C.; Lynn, A., g_mmpbsa--a GROMACS tool for high-throughput MM-PBSA calculations. \u003cem\u003eJ Chem Inf Model\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2014,\u003c/strong\u003e \u003cem\u003e54\u003c/em\u003e (7), 1951-62.\u003c/p\u003e\n\u003cp\u003e24.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Ren, X.; \u0026nbsp;Shi, Y. S.; \u0026nbsp;Zhang, Y.; \u0026nbsp;Liu, B.; \u0026nbsp;Zhang, L. H.; \u0026nbsp;Peng, Y. B.; Zeng, R., Novel Consensus Docking Strategy to Improve Ligand Pose Prediction. \u003cem\u003eJ Chem Inf Model\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2018,\u003c/strong\u003e \u003cem\u003e58\u003c/em\u003e (8), 1662-1668.\u003c/p\u003e\n\u003cp\u003e25.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Chen, Y.; \u0026nbsp;Guo, Y.; \u0026nbsp;Pan, Y.; Zhao, Z. J., Structure analysis of the receptor binding of 2019-nCoV. \u003cem\u003eBiochem Biophys Res Commun\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2020\u003c/strong\u003e.\u003c/p\u003e\n\u003cp\u003e26.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;De Vivo, M.; \u0026nbsp;Masetti, M.; \u0026nbsp;Bottegoni, G.; Cavalli, A., Role of Molecular Dynamics and Related Methods in Drug Discovery. \u003cem\u003eJ Med Chem\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2016,\u003c/strong\u003e \u003cem\u003e59\u003c/em\u003e (9), 4035-61.\u003c/p\u003e\n\u003cp\u003e27.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Omotuyi, O. I.; Ueda, H., Molecular dynamics study-based mechanism of nefiracetam-induced NMDA receptor potentiation. \u003cem\u003eComput Biol Chem\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2015,\u003c/strong\u003e \u003cem\u003e55\u003c/em\u003e, 14-22.\u003c/p\u003e\n\u003cp\u003e28.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Forman, R.; \u0026nbsp;Shah, S.; \u0026nbsp;Jeurissen, P.; \u0026nbsp;Jit, M.; Mossialos, E., COVID-19 vaccine challenges: What have we learned so far and what remains to be done? \u003cem\u003eHealth Policy\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2021,\u003c/strong\u003e \u003cem\u003e125\u003c/em\u003e (5), 553-567.\u003c/p\u003e\n\u003cp\u003e29.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Omotuyi, O.; \u0026nbsp;Olubiyi, O.; \u0026nbsp;Nash, O.; \u0026nbsp;Afolabi, E.; \u0026nbsp;Oyinloye, B.; \u0026nbsp;Fatumo, S.; \u0026nbsp;Femi-Oyewo, M.; Bogoro, S., SARS-CoV-2 Omicron spike glycoprotein receptor binding domain exhibits super-binder ability with ACE2 but not convalescent monoclonal antibody. \u003cem\u003eComput Biol Med\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2022,\u003c/strong\u003e \u003cem\u003e142\u003c/em\u003e, 105226.\u003c/p\u003e\n\u003cp\u003e30.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Paul, D.; \u0026nbsp;Pyne, N.; Paul, S., Mutation profile of SARS-CoV-2 spike protein and identification of potential multiple epitopes within spike protein for vaccine development against SARS-CoV-2. \u003cem\u003eVirusdisease\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2021,\u003c/strong\u003e \u003cem\u003e32\u003c/em\u003e (4), 703-726.\u003c/p\u003e\n\u003cp\u003e31.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Bajusz, D.; \u0026nbsp;Racz, A.; Heberger, K., Why is Tanimoto index an appropriate choice for fingerprint-based similarity calculations? \u003cem\u003eJ Cheminform\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2015,\u003c/strong\u003e \u003cem\u003e7\u003c/em\u003e, 20.\u003c/p\u003e\n\u003cp\u003e32.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Tonelli, M.; \u0026nbsp;Simone, M.; \u0026nbsp;Tasso, B.; \u0026nbsp;Novelli, F.; \u0026nbsp;Boido, V.; \u0026nbsp;Sparatore, F.; \u0026nbsp;Paglietti, G.; \u0026nbsp;Pricl, S.; \u0026nbsp;Giliberti, G.; \u0026nbsp;Blois, S.; \u0026nbsp;Ibba, C.; \u0026nbsp;Sanna, G.; \u0026nbsp;Loddo, R.; La Colla, P., Antiviral activity of benzimidazole derivatives. II. Antiviral activity of 2-phenylbenzimidazole derivatives. \u003cem\u003eBioorg Med Chem\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2010,\u003c/strong\u003e \u003cem\u003e18\u003c/em\u003e (8), 2937-53.\u003c/p\u003e\n\u003cp\u003e33.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Vitale, G.; \u0026nbsp;Corona, P.; \u0026nbsp;Loriga, M.; \u0026nbsp;Carta, A.; \u0026nbsp;Paglietti, G.; \u0026nbsp;Giliberti, G.; \u0026nbsp;Sanna, G.; \u0026nbsp;Farci, P.; \u0026nbsp;Marongiu, M. E.; La Colla, P., 5-acetyl-2-arylbenzimidazoles as antiviral agents. Part 4. \u003cem\u003eEur J Med Chem\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2012,\u003c/strong\u003e \u003cem\u003e53\u003c/em\u003e, 83-97.\u003c/p\u003e\n\u003cp\u003e34.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Tonelli, M.; \u0026nbsp;Paglietti, G.; \u0026nbsp;Boido, V.; \u0026nbsp;Sparatore, F.; \u0026nbsp;Marongiu, F.; \u0026nbsp;Marongiu, E.; \u0026nbsp;La Colla, P.; Loddo, R., Antiviral activity of benzimidazole derivatives. I. Antiviral activity of 1-substituted-2-[(benzotriazol-1/2-yl)methyl]benzimidazoles. \u003cem\u003eChem Biodivers\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2008,\u003c/strong\u003e \u003cem\u003e5\u003c/em\u003e (11), 2386-401.\u003c/p\u003e\n\u003cp\u003e35.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Kulak, J., Jr.; \u0026nbsp;Fischer, C.; \u0026nbsp;Komm, B.; Taylor, H. S., Treatment with bazedoxifene, a selective estrogen receptor modulator, causes regression of endometriosis in a mouse model. \u003cem\u003eEndocrinology\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2011,\u003c/strong\u003e \u003cem\u003e152\u003c/em\u003e (8), 3226-32.\u003c/p\u003e\n\u003cp\u003e36.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Palacios, S., Bazedoxifene acetate for the management of postmenopausal osteoporosis. \u003cem\u003eDrugs Today (Barc)\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2011,\u003c/strong\u003e \u003cem\u003e47\u003c/em\u003e (3), 187-95.\u003c/p\u003e\n\u003cp\u003e37. \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; Smetana, K., Jr.; \u0026nbsp;Rosel, D.; BrAbek, J., Raloxifene and Bazedoxifene Could Be Promising Candidates for Preventing the COVID-19 Related Cytokine Storm, ARDS and Mortality. \u003cem\u003eIn Vivo\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2020,\u003c/strong\u003e \u003cem\u003e34\u003c/em\u003e (5), 3027-3028.\u003c/p\u003e\n\u003cp\u003e38. \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; Jeon, S.; \u0026nbsp;Ko, M.; \u0026nbsp;Lee, J.; \u0026nbsp;Choi, I.; \u0026nbsp;Byun, S. Y.; \u0026nbsp;Park, S.; \u0026nbsp;Shum, D.; Kim, S., Identification of Antiviral Drug Candidates against SARS-CoV-2 from FDA-Approved Drugs. \u003cem\u003eAntimicrob Agents Chemother\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e2020,\u003c/strong\u003e \u003cem\u003e64\u003c/em\u003e (7).\u003c/p\u003e"},{"header":"Tables","content":"\u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:justify;line-height:150%;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003eTable 1.0: Compounds with high Tanimoto coefficient with benzimidazole\u003c/span\u003e\u003c/p\u003e\n\u003ctable style=\"width:282.0pt;border-collapse:collapse;border:none;\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 106pt;border: 1pt solid windowtext;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u003cstrong\u003e\u003cem\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003eCompounds\u003c/span\u003e\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: windowtext windowtext windowtext currentcolor;border-style: solid solid solid none;border-width: 1pt 1pt 1pt medium;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cstrong\u003e\u003cem\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003eVina\u003c/span\u003e\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cstrong\u003e\u003cem\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003eDock\u003c/span\u003e\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: windowtext windowtext windowtext currentcolor;border-style: solid solid solid none;border-width: 1pt 1pt 1pt medium;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cstrong\u003e\u003cem\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003eGlide XP\u003c/span\u003e\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: windowtext windowtext windowtext currentcolor;border-style: solid solid solid none;border-width: 1pt 1pt 1pt medium;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cstrong\u003e\u003cem\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003eGlide SP\u003c/span\u003e\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: windowtext windowtext windowtext currentcolor;border-style: solid solid solid none;border-width: 1pt 1pt 1pt medium;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cstrong\u003e\u003cem\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003eMean\u003c/span\u003e\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cstrong\u003e\u003cem\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003eScore\u003c/span\u003e\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 106pt;border-color: currentcolor windowtext windowtext;border-style: none solid solid;border-width: medium 1pt 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003eAlbendazole (\u003cstrong\u003e1\u003c/strong\u003e)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.20\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-2.84\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-4.27\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-4.10\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 106pt;border-color: currentcolor windowtext windowtext;border-style: none solid solid;border-width: medium 1pt 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003eetonitazene (\u003cstrong\u003e2\u003c/strong\u003e)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-6.00\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-2.48\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-4.78\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-4.42\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 106pt;border-color: currentcolor windowtext windowtext;border-style: none solid solid;border-width: medium 1pt 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003egaleterone (\u003cstrong\u003e3\u003c/strong\u003e)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-6.80\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-2.52\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-4.03\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-4.45\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 106pt;border-color: currentcolor windowtext windowtext;border-style: none solid solid;border-width: medium 1pt 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003elansoprazole (\u003cstrong\u003e4\u003c/strong\u003e)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-6.30\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.23\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.83\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.79\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 106pt;border-color: currentcolor windowtext windowtext;border-style: none solid solid;border-width: medium 1pt 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003emavatrep (\u003cstrong\u003e5\u003c/strong\u003e)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-8.00\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-3.81\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.55\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.79\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 106pt;border-color: currentcolor windowtext windowtext;border-style: none solid solid;border-width: medium 1pt 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003emebendazole (\u003cstrong\u003e6\u003c/strong\u003e)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-6.20\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-3.41\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-3.85\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-4.49\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 106pt;border-color: currentcolor windowtext windowtext;border-style: none solid solid;border-width: medium 1pt 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003eomeprazole (\u003cstrong\u003e7\u003c/strong\u003e)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.40\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.48\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.27\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.38\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 106pt;border-color: currentcolor windowtext windowtext;border-style: none solid solid;border-width: medium 1pt 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003epantoprazole (\u003cstrong\u003e8\u003c/strong\u003e)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-6.50\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-3.94\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.81\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.42\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 106pt;border-color: currentcolor windowtext windowtext;border-style: none solid solid;border-width: medium 1pt 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003erabeprazole (\u003cstrong\u003e9\u003c/strong\u003e)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.70\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.74\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.05\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.50\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 106pt;border-color: currentcolor windowtext windowtext;border-style: none solid solid;border-width: medium 1pt 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003etenatoprazole (\u003cstrong\u003e10\u003c/strong\u003e)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-6.00\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-4.03\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-3.91\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-4.65\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 106pt;border-color: currentcolor windowtext windowtext;border-style: none solid solid;border-width: medium 1pt 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003etriclabendazole (\u003cstrong\u003e11\u003c/strong\u003e)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.70\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-4.18\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-4.69\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-4.86\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 106pt;border-color: currentcolor windowtext windowtext;border-style: none solid solid;border-width: medium 1pt 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003eBazedoxifene* (\u003cstrong\u003e12\u003c/strong\u003e)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-7.40\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-5.13\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-6.20\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 14pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003e-6.24\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 106pt;border-color: currentcolor windowtext windowtext;border-style: none solid solid;border-width: medium 1pt 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 13pt;vertical-align: bottom;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 13pt;vertical-align: bottom;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 13pt;vertical-align: bottom;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 13pt;vertical-align: bottom;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 44pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 13pt;vertical-align: bottom;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u003cspan style='font-family:\"Big Caslon Medium\";'\u003eTable 2.0: MMPBSA Energy Estimation of Bazedoxifene at RBD/ACE2 interface.\u003c/span\u003e\u003c/p\u003e\n\u003ctable style=\"width:403.05pt;border-collapse:collapse;border:none;\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64.1pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 15.5pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"5\" style=\"width: 338.95pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 15.5pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;'\u003eEnergy Terms (Kj/mol)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64.1pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 19.85pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;'\u003eCompound\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 91.3pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 19.85pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;color:black;'\u003evan\u003cem\u003e\u0026nbsp;der\u003c/em\u003e Waal energy\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.6pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 19.85pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;color:black;'\u003eElectrostatic energy\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.6pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 19.85pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;color:black;'\u003ePolar solvation energy\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.2pt;border-color: currentcolor windowtext windowtext currentcolor;border-style: none solid solid none;border-width: medium 1pt 1pt medium;padding: 0in 5.4pt;height: 19.85pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;color:black;'\u003eSASA energy\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 55.2pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 19.85pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cstrong\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;color:black;'\u003eBinding energy\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64.1pt;border-color: currentcolor windowtext currentcolor currentcolor;border-style: none solid none none;border-width: medium 1pt medium medium;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 19.85pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;'\u003eBAZ\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 91.3pt;border-color: currentcolor windowtext currentcolor currentcolor;border-style: none solid none none;border-width: medium 1pt medium medium;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 19.85pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;'\u003e-128.110\u003c/span\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;color:black;'\u003e\u0026nbsp;\u0026plusmn; 20.285\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.6pt;border-color: currentcolor windowtext currentcolor currentcolor;border-style: none solid none none;border-width: medium 1pt medium medium;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 19.85pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;'\u003e-431.479\u0026nbsp;\u003c/span\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;color:black;'\u003e\u0026plusmn;\u003c/span\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;'\u003e\u0026nbsp;18.550\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.6pt;border-color: currentcolor windowtext currentcolor currentcolor;border-style: none solid none none;border-width: medium 1pt medium medium;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 19.85pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;'\u003e167.395\u0026nbsp;\u003c/span\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;color:black;'\u003e\u0026plusmn;\u003c/span\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;'\u003e\u0026nbsp;39.644\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.2pt;border-color: currentcolor windowtext currentcolor currentcolor;border-style: none solid none none;border-width: medium 1pt medium medium;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 19.85pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;'\u003e-16.984\u0026nbsp;\u003c/span\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;color:black;'\u003e\u0026plusmn; 2.664\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 55.2pt;border: medium none;padding: 0in 5.4pt;height: 19.85pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cstrong\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;'\u003e-409.178\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cspan style='font-family:\"Avenir Next Condensed\",sans-serif;color:black;'\u003e\u0026plusmn; \u0026nbsp;21.114\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp style='margin:0in;font-size:16px;font-family:\"Calibri\",sans-serif;'\u003e\u0026nbsp;\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"COVID-19, Receptor binding domain, ACE2, Bazedoxifene, SARS-COV2","lastPublishedDoi":"10.21203/rs.3.rs-1869175/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-1869175/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Coronavirus spike glycoprotein (GP)-Receptor Binding Domain (RBD) and its interaction with host receptor angiotensin converting enzyme 2 (ACE2) is one of the most structurally understood but therapeutically untapped aspect of COVID-19 pathogenesis. In this study, Fragment-based lead discovery was conducted to identify potential lead anti-SARS-COV-2 drug candidate acting at RBD/ACE2 interface. From the original 12000 chemical building blocks, benzimidazole (BAZ) scaffold was identified. Fingerprint-based similarity search of the FDA-approved drug library for BAZ-containing compounds identified 12 drugs with the benzimidazole-like substructure. When these compounds were re—docked into GP/ACE2 interface, the consensus docking identified bazedoxifene as the hit. In vitro RBD/ACE2 inhibition kinetics showed micromolar IC50 value (1.237 μM) in the presence of bazedoxifene. Molecular dynamics simulation of RBD/ACE2 in the presence BAZ resulted in loss of contact and specific hydrogen-bond interaction required for RBD/ACE2 stability. Taken together, these findings identified benzimidazole scaffold as a building block for developing novel RBD/ACE2 complex inhibitor and provided mechanistic basis for the use of bazedoxifene as a repurposable drug for the treatment of COVID-19 acting at RBD/ACE2 interface.","manuscriptTitle":"Inhibition of SARS-COV-2 Receptor-Binding domain/ACE2 by Bazedoxifene","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-07-18 21:06:36","doi":"10.21203/rs.3.rs-1869175/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"17d44a65-2255-4765-afcf-6fc2106d1a99","owner":[],"postedDate":"July 18th, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2022-08-03T13:53:51+00:00","versionOfRecord":[],"versionCreatedAt":"2022-07-18 21:06:36","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-1869175","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-1869175","identity":"rs-1869175","version":["v1"]},"buildId":"cTy_lsJlmDsVRNrSptgXS","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}