【病毒外文文獻(xiàn)】2006 Discovering Severe Acute Respiratory Syndrome Coronavirus 3CL Protease Inhibitors_ Virtual Screening, Surface Plasm

《【病毒外文文獻(xiàn)】2006 Discovering Severe Acute Respiratory Syndrome Coronavirus 3CL Protease Inhibitors_ Virtual Screening, Surface Plasm》由會(huì)員分享，可在線閱讀，更多相關(guān)《【病毒外文文獻(xiàn)】2006 Discovering Severe Acute Respiratory Syndrome Coronavirus 3CL Protease Inhibitors_ Virtual Screening, Surface Plasm（7頁(yè)珍藏版）》請(qǐng)?jiān)谘b配圖網(wǎng)上搜索。

Discovering Severe Acute Respiratory Syndrome Coronavirus 3CL Protease Inhibitors Virtual Screening Surface Plasmon Resonance and Fluorescence Resonance Energy Transfer Assays LILI CHEN 1 SHUAI CHEN 1 CHUNSHAN GUI 1 JIANHUA SHEN 1 XU SHEN 1 2 and HUALIANG JIANG 1 2 An integrated system has been developed for discovering potent inhibitors of severe acute respiratory syndrome coronavirus 3C like protease SARS CoV 3CL pro by virtual screening correlating with surface plasmon resonance SPR and fluores cence resonance energy transfer FRET technologies based assays The authors screened 81 287 small molecular com pounds against SPECS database by virtual screening 256 compounds were subsequently selected for biological evaluation Through SPR technology based assay 52 from these 256 compounds were discovered to show binding to SARS CoV 3CL pro The enzymatic inhibition activities of these 52 SARS CoV 3CL pro binders were further applied to FRET based assay and IC 50 values were determined Based on this integrated assay platform 8 new SARS CoV 3CL pro inhibitors were discovered The fact that the obtained IC 50 values for the inhibitors are in good accordance with the discovered dissociation equilibrium constants K D s assayed by SPR implied the reliability of this platform Our current work is hoped to supply a powerful approach in the discovery of potent SARS CoV 3CL pro inhibitors and the determined inhibitors could be used as possible lead compounds for further research Journal of Biomolecular Screening 2006 915 921 Key words SARS SARS CoV 3CL pro inhibitor virtual screening SPR FRET 2006 Society for Biomolecular Sciences www sbsonline org 915 T HE LAST OUTBREAK OF THE SEVERE ACUTE RESPIRATORY SYNDROME SARS epidemic in 2003 has led to thousands of lethally infected patients and hundreds of deaths 1 and the seri ous aftereffect has caused major medical and economic con cerns It has been reported that SARS coronavirus SARS CoV is responsible for SARS infection 2 3 and SARS CoV 3C like proteinase SARS CoV 3CL pro is an attractive target for the dis covery of anti SARS agents for its functional importance in the viral life cycle To date varied kinds of SARS CoV 3CL pro inhibitors were discovered 4 6 although no effective SARS CoV 3CL pro inhibitor has yet been reported to treat SARS Therefore it is still a great challenge to explore new chemical classes of SARS CoV 3CL pro inhibitors that can be possibly used in anti SARS research As has been used in the current drug discovery process by using the high performance computation technique to search the large chemical compound databases for the identification of pos sible drug candidates the virtual screening approach is a tech nology that is based on the 3D structure of the target protein It involves the rapid fitting of the chemical library members into the active sites of 3D protein structures and is critical to distin guish active from inactive substances at the primary screening stage To date there have been successful cases of virtual screen ing used in the discovery of lead compound candidates For example we recently reported that by using virtual screening technology with other methods several new small molecular specific cyclophilin A inhibitors were discovered 7 through vir tual screening with surface plasmon resonance SPR technology against a database containing structural information of more than 8000 available drugs Cinanserin SQ 10 643 a well characterized serotonin antagonist that had undergone preliminary clinical test ing in humans in the 1960s was discovered to be the SARS CoV 3CL pro inhibitor 5 Recently SPR technology has been recognized as a powerful tool in monitoring receptor ligand interactions with advantages of no labeling real time noninvasive measurements and low sample 1 Drug Discovery and Design Center State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Chinese Academy of Sciences Shanghai China 2 School of Pharmacy East China University of Science and Technology Shanghai China Received Mar 28 2006 and in revised form Jul 7 2006 Accepted for publica tion Jul 18 2006 Journal of Biomolecular Screening 11 8 2006 DOI 10 1177 1087057106293295 at TEXAS A 2006 at TEXAS A 2006 www sbsonline org 917 FIG 1 Structures of the 8 compounds screened in this study at TEXAS A 2006 Table 1 Summary of the Properties of the 8 SARS CoV 3CL pro Inhibitors No No No Rule of Molecular Hydrogen Hydrogen 5 Pass Compound Weight Bond Donors Bond Donors CLogP or Fail DC060015 558 4 2 7 6 5484 Fail DC060087 487 5 1 9 2 9486 Pass DC060159 519 4 4 7 5 6938 Fail DC060170 541 3 2 9 4 551 Pass DC060180 522 1 2 7 4 507 Pass DC060245 536 6 1 7 7 056 Fail DC060251 470 5 2 7 5 7126 Pass DC060256 420 3 1 5 3 2525 Pass FIG 2 The binding model of the 8 inhibitors against SARS CoV 3CL pro in the active site DC060251 is rendered in sticks whereas others are shown in lines FIG 3 Schematic representation of SARS CoV 3CL pro interaction with DC060251 DC060251 and protein residues are rendered in ball and stick and sticks respectively at TEXAS A 2006 www sbsonline org 919 FIG 4 Structures of 3 compounds A B C from MDL Drug Data Report MDDR Comprehensive Medicinal Chemistry CMC and China Natural Products Database CNPD databases DC060251 shows 90 structural similarity to them Table 2 Summary of Concentration of Inhibitor Producing 50 Inhibition IC 50 Values and Equilibrium Dissociation Constants K D s for SARS CoV 3CL pro Inhibitors Molecular Compound Weight Da IC 50 M K D M DC060015 558 4 49 96 4 62 18 60 1 18 DC060087 487 5 80 46 2 07 42 50 2 10 DC060159 519 4 42 19 3 02 19 14 2 03 DC060170 541 3 6 86 0 91 4 23 0 81 DC060180 522 1 9 19 2 34 6 50 0 23 DC060245 536 6 21 49 2 97 23 91 5 63 DC060251 470 5 9 56 1 75 4 78 0 29 DC060256 420 3 56 24 3 48 49 91 3 98 at TEXAS A 113 701 702 2 Drosten C Preiser W Gunther S Schmitz H Doerr HW Severe acute res piratory syndrome identification of the etiological agent Trends Mol Med 2003 9 325 327 3 Fouchier RA Kuiken T Schutten M van Amerongen G van Doornum GJ van den Hoogen BG et al Koch s postulates fulfilled for SARS virus Nature 2003 423 240 4 Bacha U Barrila J Velazquez Campoy A Leavitt SA Freire E Identification of novel inhibitors of the SARS coronavirus main protease 3CLpro Biochemistry 2004 43 4906 4912 5 Chen L Gui C Luo X Yang Q Gunther S Scandella E et al Cinanserin is an inhibitor of the 3C like proteinase of severe acute respiratory syndrome coronavirus and strongly reduces virus replication in vitro J Virol 2005 79 7095 7103 Chen et al 920 www sbsonline org Journal of Biomolecular Screening 11 8 2006 FIG 6 Correlation between the concentration of inhibitor produc ing 50 inhibition IC 50 and equilibrium dissociation constants K D for SARS CoV 3CL pro inhibitors IC 50 values were obtained from the fluorescence resonance energy transfer FRET based assay and K D values from biosensor measurements The symbol z represents the compounds with correlation FIG 5 Dose response inhibition and kinetic assay curves for the 3 compounds from the screening procedure IC 50 values were extracted from the curves using nonlinear regression analysis and dissociation equilibrium constant K D values were analyzed using a global data analysis program of Biacore 3000 A B C represent the dose response inhibition curves Inset the responding binding curve of the compounds DC060170 DC060180 DC060251 respectively at TEXAS A 11 1445 1453 7 Li J Chen J Gui C Zhang L Qin Y Xu Q et al Discovering novel chem ical inhibitors of human cyclophilin A virtual screening synthesis and bioassay Bioorg Med Chem 2006 14 2209 2224 8 Cooper MA Optical biosensors in drug discovery Nat Rev Drug Discov 2002 1 515 528 9 Markgren PO Hamalainen M Danielson UH Screening of compounds interacting with HIV 1 proteinase using optical biosensor technology Anal Biochem 1998 265 340 350 10 Du L Zhang Z Luo X Chen K Shen X Jiang H Binding investigation of human 5 lipoxygenase with its inhibitors by SPR technology correlating with molecular docking simulation J Biochem Tokyo 2006 139 715 723 11 Ye F Zhang ZS Luo HB Shen JH Chen KX Shen X et al The dipeptide H Trp Glu OH shows highly antagonistic activity against PPARgamma bioassay with molecular modeling simulation Chembiochem 2006 7 74 82 12 Chen S Chen LL Luo HB Sun T Chen J Ye F et al Enzymatic activity characterization of SARS coronavirus 3C like protease by fluorescence res onance energy transfer technique Acta Pharmacol Sin 2005 26 99 106 13 Chen S Chen L Tan J Chen J Du L Sun T et al Severe acute respira tory syndrome coronavirus 3C like proteinase N terminus is indispens able for proteolytic activity but not for enzyme dimerization Biochemical and thermodynamic investigation in conjunction with molecular dynam ics simulations J Biol Chem 2005 280 164 173 14 Sun H Luo H Yu C Sun T Chen J Peng S et al Molecular cloning expression purification and mass spectrometric characterization of 3C like protease of SARS coronavirus Protein Expr Purif 2003 32 302 308 15 Doman TN McGovern SL Witherbee BJ Kasten TP Kurumbail R Stallings WC et al Molecular docking and high throughput screening for novel inhibitors of protein tyrosine phosphatase 1B J Med Chem 2002 45 2213 2221 16 Ewing TJ Makino S Skillman AG Kuntz ID DOCK 4 0 search strate gies for automated molecular docking of flexible molecule databases J Comput Aided Mol Des 2001 15 411 428 17 Cornell WD Cieplak D Bayly CI Gould IR Merz KM Ferguson DM et al A Second generation force field for the simulation of proteins nucleic acids and organic molecules J Am Chem SOC 1995 117 5179 5197 18 Gasteiger J Marsili M Iterative partial equalization of orbital elec tronegativity A rapid access to atomic charges Tetrahedron 1980 36 3219 3228 19 Purcell WP Singer JA Brief review and table of semiempirical parameters used in the H ckel molecular orbital method J Chem Eng Data 1967 12 235 246 20 Clark R Strizhev A Leonard J Blake J Matthew J Consensus scoring for ligand protein interactions J Mol Graph Model 2002 20 281 295 21 Zheng S Luo X Chen G Zhu W Shen J Chen K et al A new rapid and effective chemistry space filter in recognizing a druglike database J Chem Inf Model 2005 45 856 862 22 Johnsson B Lofas S Lindquist G Immobilization of proteins to a carboxymethyldextran modified gold surface for biospecific interaction analysis in surface plasmon resonance sensors Anal Biochem 1991 198 268 277 23 Xiong B Gui CS Xu XY Luo C Chen J Luo HB et al A 3D model of SARS CoV 3CL proteinase and its inhibitors design by virtual screening Acta Pharmacol Sin 2003 24 497 504 24 Wang S Milne GW Yan X Posey IJ Nicklaus MC Graham L et al Discovery of novel non peptide HIV 1 protease inhibitors by pharma cophore searching J Med Chem 1996 39 2047 2054 25 Stahmann MA Wolff I Studies on 4 hydroxycoumarins I The synthesis of 4 hydroxycoumarins J Am Chem Soc 1943 65 2285 2287 26 Sengupta P Sen M Karuri P Structure of gerberinol novel dimethyl dicoumarol from gerbera lanuginosa benth J Indian Chem Soc 1985 62 916 919 27 Luo H Wu D Shen C Chen K Shen X Jiang H Severe acute respira tory syndrome coronavirus membrane protein interacts with nucleocapsid protein mostly through their carboxyl termini by electrostatic attraction Int J Biochem Cell Biol 2006 38 589 599 28 Luo H Chen Q Chen J Chen K Shen X Jiang H The nucleocapsid protein of SARS coronavirus has a high binding affinity to the human cellular heterogeneous nuclear ribonucleoprotein A1 FEBS Lett 2005 579 2623 2628 29 Kao RY To AP Ng LW Tsui WH Lee TS Tsoi HW et al Characterization of SARS CoV main protease and identification of biologically active small molecule inhibitors using a continuous fluorescence based assay FEBS Lett 2004 576 325 330 30 Gao Z Luo H Chen L Shen J Chen K Jiang H et al Determining PPAR ligand binding affinity using fluorescent assay with cis parinaric acid as a probe Science in China Ser B Chemistry 2005 48 1 10 Address reprint requests to Xu Shen Drug Discovery and Design Center State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Chinese Academy of Sciences Shanghai 201203 E mail xshen or hljiang SARS Coronavirus 3CL Protease Inhibitors Journal of Biomolecular Screening 11 8 2006 www sbsonline org 921 at TEXAS A M INTL UNIV on March 16 Downloaded from