{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,27]],"date-time":"2026-03-27T03:23:46Z","timestamp":1774581826168,"version":"3.50.1"},"reference-count":61,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2025,5,13]],"date-time":"2025-05-13T00:00:00Z","timestamp":1747094400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>Rho-associated protein kinase 2 (ROCK2) is a serine\/threonine kinase that is crucial for regulating various physiological processes and is part of the Rho-associated coiled-coil kinase family. The dysregulation of ROCK2 has been associated with a range of diseases, making it a promising target for therapy. In this study, a chemical feature-based pharmacophore model was developed on the co-crystal ligand (5YS) of ROCK2 to conduct the virtual screening of ZINC database, resulting in 4809 hits that were further subjected to molecular docking to find the binding affinities with ROCK2 protein. The binding affinities of the hits were analyzed and compounds in the range of -11.55 to -9.91 kcal\/mol were selected for further analysis. The ADMET analysis identified two promising compounds, whose binding stability with the ROCK2 protein was further evaluated using molecular dynamics (MD) simulations. Simulation results revealed that the selected compounds remained closely bound to protein indicating that they can act as lead compounds to control the biological activity of ROCK2. However, further in vitro investigation is required to test the biological efficacy of the reported compounds.<\/jats:p>","DOI":"10.1371\/journal.pone.0323781","type":"journal-article","created":{"date-parts":[[2025,5,13]],"date-time":"2025-05-13T18:03:21Z","timestamp":1747159401000},"page":"e0323781","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":2,"title":["Discovery of ROCK2 inhibitors through computational screening of ZINC database: Integrating pharmacophore modeling, molecular docking, and MD simulations"],"prefix":"10.1371","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-2565-6573","authenticated-orcid":true,"given":"Abdullah R.","family":"Alanzi","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0456-3196","authenticated-orcid":true,"given":"Abdelaaty A.","family":"Shahat","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-8340-4395","authenticated-orcid":true,"given":"Bayan Abdullah","family":"Alhaidhal","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Raghad Mohammad","family":"Aloatibi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2025,5,13]]},"reference":[{"issue":"1","key":"pone.0323781.ref001","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1186\/s40478-017-0447-3","article-title":"Rho-associated protein kinase 2 (ROCK2): a new target of autoimmunity in paraneoplastic encephalitis","volume":"5","author":"S Popkirov","year":"2017","journal-title":"Acta Neuropathol Commun"},{"key":"pone.0323781.ref002","doi-asserted-by":"crossref","DOI":"10.4161\/sgtp.29846","article-title":"Rho-associated coiled-coil containing kinases (ROCK): structure, regulation, and functions","volume":"5","author":"L Julian","year":"2014","journal-title":"Small GTPases"},{"issue":"2","key":"pone.0323781.ref003","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1247\/csf.12018","article-title":"Distinct distribution and localization of Rho-kinase in mouse epithelial, muscle and neural tissues","volume":"37","author":"M Iizuka","year":"2012","journal-title":"Cell Struct Funct"},{"issue":"4","key":"pone.0323781.ref004","doi-asserted-by":"crossref","first-page":"1074","DOI":"10.1124\/pr.115.010595","article-title":"Rho kinases in health and disease: from basic science to translational research","volume":"67","author":"G Loirand","year":"2015","journal-title":"Pharmacol Rev"},{"key":"pone.0323781.ref005","doi-asserted-by":"crossref","DOI":"10.12688\/f1000research.7909.1","article-title":"Rho, ROCK and actomyosin contractility in metastasis as drug targets","volume":"5","author":"I Rodriguez-Hernandez","year":"2016","journal-title":"F1000Res"},{"issue":"7","key":"pone.0323781.ref006","first-page":"2632","article-title":"Significant association of Rho\/ROCK pathway with invasion and metastasis of bladder cancer","volume":"9","author":"T Kamai","year":"2003","journal-title":"Clin Cancer Res"},{"key":"pone.0323781.ref007","doi-asserted-by":"crossref","first-page":"198","DOI":"10.3389\/fnins.2015.00198","article-title":"Chatting with the neighbors: crosstalk between Rho-kinase (ROCK) and other signaling pathways for treatment of neurological disorders","volume":"9","author":"N Hensel","year":"2015","journal-title":"Front Neurosci"},{"issue":"2","key":"pone.0323781.ref008","doi-asserted-by":"crossref","DOI":"10.1152\/ajpheart.00327.2011","article-title":"Rho-kinase: important new therapeutic target in cardiovascular diseases","volume":"301","author":"K Satoh","year":"2011","journal-title":"Am J Physiol Heart Circ Physiol"},{"issue":"5","key":"pone.0323781.ref009","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1038\/nrd1719","article-title":"Rho kinase, a promising drug target for neurological disorders","volume":"4","author":"BK Mueller","year":"2005","journal-title":"Nat Rev Drug Discov"},{"issue":"4","key":"pone.0323781.ref010","doi-asserted-by":"crossref","first-page":"455","DOI":"10.2174\/1389450117666160401123825","article-title":"ROCK in CNS: different roles of isoforms and therapeutic target for neurodegenerative disorders","volume":"18","author":"C-M Chong","year":"2017","journal-title":"Curr Drug Targets"},{"issue":"3","key":"pone.0323781.ref011","doi-asserted-by":"crossref","first-page":"605","DOI":"10.2147\/TCRM.S2907","article-title":"The therapeutic effects of Rho-ROCK inhibitors on CNS disorders","volume":"4","author":"T Kubo","year":"2008","journal-title":"Ther Clin Risk Manag"},{"issue":"9","key":"pone.0323781.ref012","doi-asserted-by":"crossref","first-page":"697","DOI":"10.3109\/00207454.2013.877014","article-title":"Intracranial injection of PEG-PEI\/ROCK II-siRNA improves cognitive impairment in a mouse model of Alzheimer\u2019s disease","volume":"124","author":"X Wen","year":"2014","journal-title":"Int J Neurosci"},{"key":"pone.0323781.ref013","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1016\/j.nbd.2014.10.018","article-title":"Post-status epilepticus treatment with the cannabinoid agonist WIN 55,212-2 prevents chronic epileptic hippocampal damage in rats","volume":"73","author":"R Di Maio","year":"2015","journal-title":"Neurobiol Dis"},{"issue":"1","key":"pone.0323781.ref014","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1038\/s41467-019-13781-3","article-title":"ROCK inhibitors upregulate the neuroprotective Parkin-mediated mitophagy pathway","volume":"11","author":"N Moskal","year":"2020","journal-title":"Nat Commun"},{"key":"pone.0323781.ref015","first-page":"1","article-title":"Fasudil attenuates aggregation of \u03b1-synuclein in models of Parkinson\u2019s disease","volume":"4","author":"L Tatenhorst","year":"2016","journal-title":"J Neurochem"},{"issue":"6","key":"pone.0323781.ref016","doi-asserted-by":"crossref","first-page":"2269","DOI":"10.1021\/acs.jmedchem.5b00683","article-title":"Rho Kinase (ROCK) inhibitors and their therapeutic potential","volume":"59","author":"Y Feng","year":"2016","journal-title":"J Med Chem"},{"key":"pone.0323781.ref017","doi-asserted-by":"crossref","first-page":"1323","DOI":"10.1016\/j.drudis.2013.09.010","article-title":"Advances in the development of Rho-associated protein kinase (ROCK) inhibitors","volume":"18","author":"P Pan","year":"2013","journal-title":"Drug Discov Today"},{"issue":"10","key":"pone.0323781.ref018","doi-asserted-by":"crossref","first-page":"2383","DOI":"10.1016\/j.bmcl.2016.03.113","article-title":"A review on ROCK-II inhibitors: From molecular modelling to synthesis","volume":"26","author":"S Shah","year":"2016","journal-title":"Bioorg Med Chem Lett"},{"issue":"1","key":"pone.0323781.ref019","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1046\/j.1471-4159.2002.00801.x","article-title":"Inhibition of rho-kinase-induced myristoylated alanine-rich C kinase substrate (MARCKS) phosphorylation in human neuronal cells by H-1152, a novel and specific Rho-kinase inhibitor","volume":"81","author":"M Ikenoya","year":"2002","journal-title":"J Neurochem"},{"key":"pone.0323781.ref020","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1016\/S0163-7258(02)00191-2","article-title":"The novel and specific Rho-kinase inhibitor (S)-(+)-2-methyl-1-[(4-methyl-5-isoquinoline)sulfonyl]-homopiperazine as a probing molecule for Rho-kinase-involved pathway","volume":"93","author":"Y Sasaki","year":"2002","journal-title":"Pharmacol Ther"},{"issue":"6","key":"pone.0323781.ref021","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1080\/1475636021000005659","article-title":"Rho-kinase inhibitors: pharmacomodulations on the lead compound Y-32885","volume":"17","author":"C Log\u00e9","year":"2002","journal-title":"J Enzyme Inhib Med Chem"},{"issue":"3","key":"pone.0323781.ref022","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1093\/jb\/mvj172","article-title":"Structural basis for induced-fit binding of Rho-kinase to the inhibitor Y-27632","volume":"140","author":"H Yamaguchi","year":"2006","journal-title":"J Biochem"},{"issue":"3","key":"pone.0323781.ref023","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1016\/j.jbiotec.2009.12.003","article-title":"Integration of virtual screening with high-throughput screening for the identification of novel Rho-kinase I inhibitors","volume":"145","author":"L-L Gong","year":"2010","journal-title":"J Biotechnol"},{"issue":"6","key":"pone.0323781.ref024","doi-asserted-by":"crossref","first-page":"1511","DOI":"10.1039\/c3mb00016h","article-title":"Discovery of Rho-kinase inhibitors by docking-based virtual screening","volume":"9","author":"M Shen","year":"2013","journal-title":"Mol Biosyst"},{"issue":"20","key":"pone.0323781.ref025","doi-asserted-by":"crossref","first-page":"1755","DOI":"10.2174\/1568026618666181025114157","article-title":"Advancements in docking and molecular dynamics simulations towards ligand-receptor interactions and structure-function relationships","volume":"18","author":"AAT Naqvi","year":"2018","journal-title":"Curr Top Med Chem"},{"key":"pone.0323781.ref026","doi-asserted-by":"crossref","DOI":"10.1093\/nar\/gkw287","article-title":"Pharmit: interactive exploration of chemical space","volume":"44","author":"J Sunseri","year":"2016","journal-title":"Nucleic Acids Res"},{"issue":"6","key":"pone.0323781.ref027","doi-asserted-by":"crossref","first-page":"2663","DOI":"10.1080\/07391102.2020.1841677","article-title":"Multi-conformational frame from molecular dynamics as a structure-based pharmacophore model for mapping, screening and identifying ligands against PPAR-\u03b3: a new protocol to develop promising candidates","volume":"40","author":"P Prabitha","year":"2022","journal-title":"J Biomol Struct Dyn"},{"issue":"4","key":"pone.0323781.ref028","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1016\/j.ddtec.2004.11.007","article-title":"Lead- and drug-like compounds: the rule-of-five revolution","volume":"1","author":"CA Lipinski","year":"2004","journal-title":"Drug Discov Today Technol"},{"key":"pone.0323781.ref029","unstructured":"LigPrep, LigPrep. 2018, Schr\u00f6dinger, LLC."},{"issue":"8","key":"pone.0323781.ref030","doi-asserted-by":"crossref","first-page":"2553","DOI":"10.1021\/ct300203w","article-title":"Improving the prediction of absolute solvation free energies using the next generation OPLS force field","volume":"8","author":"D Shivakumar","year":"2012","journal-title":"J Chem Theory Comput"},{"key":"pone.0323781.ref031","first-page":"2017","volume-title":"Schr\u00f6dinger","author":"LJSS Schr\u00f6dinger","year":"2017"},{"key":"pone.0323781.ref032","doi-asserted-by":"crossref","DOI":"10.1007\/978-1-0716-0892-0_14","volume-title":"Protein structure modeling with MODELLER","author":"B Webb","year":"2021"},{"issue":"3","key":"pone.0323781.ref033","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1007\/s10822-013-9643-9","article-title":"Effects of histidine protonation and rotameric states on virtual screening of M. tuberculosis RmlC","volume":"27","author":"MO Kim","year":"2013","journal-title":"J Comput Aided Mol Des"},{"issue":"7","key":"pone.0323781.ref034","doi-asserted-by":"crossref","first-page":"1739","DOI":"10.1021\/jm0306430","article-title":"Glide: a new approach for rapid, accurate docking and scoring. 1. Method and assessment of docking accuracy","volume":"47","author":"RA Friesner","year":"2004","journal-title":"J Med Chem"},{"issue":"5","key":"pone.0323781.ref035","doi-asserted-by":"crossref","first-page":"1663","DOI":"10.1093\/bib\/bbz103","article-title":"Computational\/in silico methods in drug target and lead prediction","volume":"21","author":"FE Agamah","year":"2020","journal-title":"Brief Bioinform"},{"key":"pone.0323781.ref036","doi-asserted-by":"crossref","first-page":"1177932219865533","DOI":"10.1177\/1177932219865533","article-title":"Homology modelling and molecular docking studies of selected substituted benzo[d]imidazol-1-yl)methyl)benzimidamide scaffolds on Plasmodium falciparum adenylosuccinate lyase receptor","volume":"13","author":"GO Oduselu","year":"2019","journal-title":"Bioinform Biol Insights"},{"issue":"1","key":"pone.0323781.ref037","first-page":"1","article-title":"In silico design and pharmacokinetics investigation of some novel hepatitis C virus NS5B inhibitors: pharmacoinformatics approach","volume":"46","author":"S Ejeh","year":"2022","journal-title":"J Hepatol"},{"key":"pone.0323781.ref038","doi-asserted-by":"crossref","unstructured":"Jorgensen W, Chandrasekhar J, Madura JD, Impey RW, Klein ML. 1983;79:926.","DOI":"10.1063\/1.445869"},{"key":"pone.0323781.ref039","doi-asserted-by":"crossref","unstructured":"Qureshi KA, Al Nasr I, Koko WS, Khan TA, Qaiser Fatmi M, Imtiaz M, et al. In Vitro and In Silico Approaches for the Antileishmanial Activity Evaluations of Actinomycins Isolated from Novel Streptomyces smyrnaeus Strain UKAQ_23. 2021;10(8):887.","DOI":"10.3390\/antibiotics10080887"},{"issue":"12","key":"pone.0323781.ref040","doi-asserted-by":"crossref","first-page":"1463","DOI":"10.1002\/(SICI)1096-987X(199709)18:12<1463::AID-JCC4>3.0.CO;2-H","article-title":"LINCS: A linear constraint solver for molecular simulations","volume":"18","author":"B Hess","year":"1997","journal-title":"J Comput Chem"},{"key":"pone.0323781.ref041","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1080\/08927029108022142","article-title":"Generalized verlet algorithm for efficient molecular dynamics simulations with long-range interactions","volume":"6","author":"H Grubm\u00fcller","year":"1991","journal-title":"Molecular Simulation"},{"issue":"19","key":"pone.0323781.ref042","doi-asserted-by":"crossref","first-page":"8577","DOI":"10.1063\/1.470117","article-title":"A smooth particle mesh Ewald method","volume":"103","author":"U Essmann","year":"1995","journal-title":"J Chem Physics"},{"issue":"1","key":"pone.0323781.ref043","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1002\/pro.3923","article-title":"The Bio3D packages for structural bioinformatics","volume":"30","author":"BJ Grant","year":"2021","journal-title":"Protein Sci"},{"issue":"25","key":"pone.0323781.ref044","doi-asserted-by":"crossref","first-page":"2135","DOI":"10.1002\/jcc.23354","article-title":"CHARMM36 all-atom additive protein force field: validation based on comparison to NMR data","volume":"34","author":"J Huang","year":"2013","journal-title":"J Comput Chem"},{"issue":"15","key":"pone.0323781.ref045","doi-asserted-by":"crossref","first-page":"3993","DOI":"10.1080\/07391102.2017.1404935","article-title":"In-Silico molecular docking and simulation studies on novel chalcone and flavone hybrid derivatives with 1, 2, 3-triazole linkage as vital inhibitors of Plasmodium falciparum dihydroorotate dehydrogenase","volume":"36","author":"M Thillainayagam","year":"2018","journal-title":"J Biomol Struct Dyn"},{"issue":"1","key":"pone.0323781.ref046","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1016\/j.jsps.2015.02.008","article-title":"Synthesis, molecular properties, toxicity and biological evaluation of some new substituted imidazolidine derivatives in search of potent anti-inflammatory agents","volume":"24","author":"A Husain","year":"2016","journal-title":"Saudi Pharm J"},{"issue":"3","key":"pone.0323781.ref047","article-title":"In silico study, synthesis, and evaluation of the antimalarial activity of hybrid dimethoxy pyrazole 1, 3, 5\u2010triazine derivatives","volume":"35","author":"P Gogoi","year":"2021","journal-title":"J Chem"},{"issue":"1","key":"pone.0323781.ref048","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1007\/s11030-018-9867-7","article-title":"Design, synthesis, and in silico studies of novel eugenyloxy propanol azole derivatives having potent antinociceptive activity and evaluation of their \u03b2-adrenoceptor blocking property","volume":"23","author":"S Behrouz","year":"2019","journal-title":"Mol Divers"},{"issue":"4","key":"pone.0323781.ref049","doi-asserted-by":"crossref","first-page":"1518","DOI":"10.1021\/acs.jctc.7b00028","article-title":"How Molecular Size Impacts RMSD Applications in Molecular Dynamics Simulations","volume":"13","author":"K Sargsyan","year":"2017","journal-title":"J Chem Theory Comput"},{"issue":"3","key":"pone.0323781.ref050","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0119264","article-title":"Automatic identification of mobile and rigid substructures in molecular dynamics simulations and fractional structural fluctuation analysis","volume":"10","author":"L Mart\u00ednez","year":"2015","journal-title":"PLoS One"},{"issue":"2","key":"pone.0323781.ref051","doi-asserted-by":"crossref","first-page":"537","DOI":"10.1007\/s11030-015-9650-y","article-title":"High-throughput screening against ~6.1 million structurally diverse, lead-like compounds to discover novel ROCK inhibitors for cerebral injury recovery","volume":"20","author":"H Gong","year":"2016","journal-title":"Mol Divers"},{"issue":"3","key":"pone.0323781.ref052","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1039\/c2mb25408e","article-title":"Discovery and optimization of triazine derivatives as ROCK1 inhibitors: molecular docking, molecular dynamics simulations and free energy calculations","volume":"9","author":"M Shen","year":"2013","journal-title":"Mol Biosyst"},{"issue":"23","key":"pone.0323781.ref053","doi-asserted-by":"crossref","first-page":"12916","DOI":"10.3390\/ijms222312916","article-title":"Rho-Kinase as a target for cancer therapy and its immunotherapeutic potential","volume":"22","author":"S Kim","year":"2021","journal-title":"Int J Mol Sci"},{"issue":"3","key":"pone.0323781.ref054","doi-asserted-by":"crossref","first-page":"749","DOI":"10.18596\/jotcsa.927426","article-title":"Pharmacophore modeling in drug discovery: methodology and current status","volume":"8","author":"MT Muhammed","year":"2021","journal-title":"J Turkish Chem Soc Sec A Chem"},{"key":"pone.0323781.ref055","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/B978-0-12-820546-4.00006-4","article-title":"Molecular docking analysis: basic technique to predict drug-receptor interactions","author":"O Silakari","year":"2021","journal-title":"Concepts Exp Protoc Model Inform Drug Des"},{"issue":"18","key":"pone.0323781.ref056","doi-asserted-by":"crossref","first-page":"4574","DOI":"10.3390\/ijms20184574","article-title":"Key topics in molecular docking for drug design","volume":"20","author":"PHM Torres","year":"2019","journal-title":"Int J Mol Sci"},{"key":"pone.0323781.ref057","first-page":"1","article-title":"Molecular docking in modern drug discovery: principles and recent applications","volume":"2","author":"A Sethi","year":"2019","journal-title":"Drug Discov Dev New Adv"},{"issue":"7","key":"pone.0323781.ref058","doi-asserted-by":"crossref","first-page":"595","DOI":"10.3851\/IMP3044","article-title":"Evaluation of antiviral drug synergy in an infectious HCV system","volume":"21","author":"B Lin","year":"2016","journal-title":"Antivir Ther"},{"issue":"1","key":"pone.0323781.ref059","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1471-2377-9-S1-S1","article-title":"Addme\u2013avoiding drug development mistakes early: central nervous system drug discovery perspective","volume":"9","author":"K Tsaioun","year":"2009","journal-title":"BMC Neurol"},{"issue":"5054","key":"pone.0323781.ref060","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1126\/science.1566067","article-title":"Alzheimer\u2019s disease: the amyloid cascade hypothesis","volume":"256","author":"JA Hardy","year":"1992","journal-title":"Science"},{"issue":"1","key":"pone.0323781.ref061","doi-asserted-by":"crossref","first-page":"71","DOI":"10.3390\/pr9010071","article-title":"Molecular dynamics simulations in drug discovery and pharmaceutical development","volume":"9","author":"OMH Salo-Ahen","year":"2020","journal-title":"Processes"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0323781","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,5,13]],"date-time":"2025-05-13T18:03:40Z","timestamp":1747159420000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0323781"}},"subtitle":[],"editor":[{"given":"Oluwafemi Adeleke","family":"Ojo,","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2025,5,13]]},"references-count":61,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2025,5,13]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0323781","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,5,13]]}}}