{"corpus_id":23661802,"paper_sha":"0a6d9914d448025f15bf9d7d6d43b4d892ce2755","doi":"10.1177/1087057108329452","arxiv_id":null,"pmid":19196697,"pmcid":null,"mag_id":2125829020,"dblp_id":null,"acl_id":null,"title":"Comparison of 3 Cytotoxicity Screening Assays and Their Application to the Selection of Novel Antibacterial Hits","year":2009,"publication_date":"2009-02-01","venue":"Journal of biomolecular screening","journal":{"name":"Journal of Biomolecular Screening","pages":"142 - 150","volume":"14"},"journal_issn":null,"journal_title":null,"publication_types":["JournalArticle","Study"],"pubmed_pub_types":["Comparative Study","Evaluation Study","Journal Article"],"s2_fields_of_study":["Biology","Medicine","Chemistry"],"reference_count":30,"citation_count":26,"influential_citation_count":1,"is_open_access":false,"arxiv_categories":null,"arxiv_license":null,"arxiv_journal_ref":null,"mesh_headings":[{"d":"Animals","mj":false,"ui":"D000818"},{"d":"Anti-Bacterial Agents","mj":false,"qs":[{"q":"chemistry","mj":false,"ui":"Q000737"},{"q":"isolation & purification","mj":true,"ui":"Q000302"},{"q":"pharmacology","mj":false,"ui":"Q000494"}],"ui":"D000900"},{"d":"CHO Cells","mj":false,"ui":"D016466"},{"d":"Cell Proliferation","mj":false,"qs":[{"q":"drug effects","mj":false,"ui":"Q000187"}],"ui":"D049109"},{"d":"Cricetinae","mj":false,"ui":"D006224"},{"d":"Cricetulus","mj":false,"ui":"D003412"},{"d":"Cytotoxins","mj":false,"qs":[{"q":"analysis","mj":true,"ui":"Q000032"},{"q":"chemistry","mj":false,"ui":"Q000737"},{"q":"pharmacology","mj":false,"ui":"Q000494"}],"ui":"D003603"},{"d":"Drug Evaluation, Preclinical","mj":false,"qs":[{"q":"methods","mj":true,"ui":"Q000379"}],"ui":"D004353"},{"d":"Humans","mj":false,"ui":"D006801"},{"d":"Jurkat Cells","mj":false,"ui":"D019169"},{"d":"Microbial Sensitivity Tests","mj":false,"qs":[{"q":"methods","mj":false,"ui":"Q000379"}],"ui":"D008826"},{"d":"Microbial Viability","mj":false,"qs":[{"q":"drug effects","mj":false,"ui":"Q000187"}],"ui":"D050296"},{"d":"Models, Biological","mj":false,"ui":"D008954"},{"d":"Structure-Activity Relationship","mj":false,"ui":"D013329"}],"chemicals":[{"n":"Anti-Bacterial Agents","ui":"D000900","reg":"0"},{"n":"Cytotoxins","ui":"D003603","reg":"0"}],"comments_corrections":null,"source_flags":5,"s2_open_access_pdf_url":null,"s2_open_access_landing_url":null,"s2_open_access_license":null,"s2_open_access_status":null,"pmc_open_access_pdf_url":null,"pmc_open_access_landing_url":null,"pmc_open_access_license":null,"pmc_open_access_status":null,"unpaywall_open_access_pdf_url":null,"unpaywall_open_access_landing_url":null,"unpaywall_open_access_license":null,"unpaywall_open_access_status":null,"abstract":"Cytotoxicity screening of new chemical entities in antibacterial drug discovery discerns between cytotoxic and antimicrobial activity, thus providing predictive evidence for selective toxicity. The objective of this study was to evaluate 3 cytotoxicity assays in identifying novel antibacterial hits with desired safety margins. The endpoints in assays comprised adenylate kinase (AK) release rate as an indicator of membrane rupture (Toxilight™), intracellular adenosine triphosphate (CellTiter-Glo™), and reduction of resazurin (CellTiter-Blue™) both as indicators of cell metabolic activity. In the CellTiter-Glo™ and the CellTiter-Blue™ assays, 7 of 8 selected compounds showed cytotoxicity, whereas in the Toxilight™ assay, 3 of 8 compounds significantly reduced cell viability in the ChoK1 and the JurkatE6.1 cell line. The CellTiter-Glo™ assay proved to be the most sensitive among the evaluated assays, and excellent Z′ values were obtained in the 96-well plate (Z′ > 0.83). The CellTiter-Glo™ assay was clearly superior to the CellTiter-Blue™ and the Toxilight™ assay for the initial cytotoxicity screening. Moreover, the application of the CellTiter-Glo™ assay to determine mammalian cell toxicity versus the antibacterial effect ratio contributed to early identification of antibacterial hits with desired safety margins. The chemical structures of these novel antibacterial hits are disclosed herein. (Journal of Biomolecular Screening 2009:142-150)","claims":[{"public_id":"cl_9b3295ee279d4059841905407e480b9d","status":"active","text":"CellTiter-Glo and CellTiter-Blue detected cytotoxicity in 7 of 8 selected compounds, whereas Toxilight significantly reduced cell viability for 3 of 8 compounds in ChoK1 and JurkatE6.1 cells.","confidence":0.95,"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/claims/cl_9b3295ee279d4059841905407e480b9d"},{"public_id":"cl_c7fe1ab21f2d88d364d7973bd02d9cab","status":"active","text":"CellTiter-Glo produced excellent Z′ values in the 96-well plate format, with Z′ greater than 0.83.","confidence":0.94,"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/claims/cl_c7fe1ab21f2d88d364d7973bd02d9cab"},{"public_id":"cl_9a95eaba70c77e4fb67824b032818f6f","status":"active","text":"CellTiter-Glo was the most sensitive of the evaluated cytotoxicity assays and outperformed CellTiter-Blue and Toxilight for initial cytotoxicity screening.","confidence":0.98,"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/claims/cl_9a95eaba70c77e4fb67824b032818f6f"},{"public_id":"cl_a7247e70f04332cd81c08394aa8e304d","status":"active","text":"Using CellTiter-Glo to assess the mammalian cell toxicity to antibacterial effect ratio enabled early identification of antibacterial hits with desired safety margins.","confidence":0.93,"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/claims/cl_a7247e70f04332cd81c08394aa8e304d"}],"concepts":[{"public_id":"co_25a9feb2d745090c037c2835d1075285","status":"active","name":"cytotoxicity screening assays","description":"Assays used to distinguish cytotoxic activity from antimicrobial activity during antibacterial drug discovery.","types":["assay class"],"aliases":["cytotoxicity screening"],"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/concepts/co_25a9feb2d745090c037c2835d1075285"},{"public_id":"co_384cd1c1dbb95e4affd1f070fc813f8d","status":"active","name":"adenylate kinase","description":"An enzyme whose release is used here as a marker of membrane rupture.","types":["biomolecule","marker"],"aliases":["AK"],"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/concepts/co_384cd1c1dbb95e4affd1f070fc813f8d"},{"public_id":"co_4a70c919dda781ac7512200ce4206b8f","status":"active","name":"96-well plate","description":"A multiwell assay plate format used for cytotoxicity screening.","types":["laboratory plate"],"aliases":["96-well plate format"],"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/concepts/co_4a70c919dda781ac7512200ce4206b8f"},{"public_id":"co_63339ab48ab84253833955f9a8049f62","status":"active","name":"JurkatE6.1","description":"A human T-cell leukemia cell line used in the cytotoxicity assays.","types":["cell line"],"aliases":["Jurkat E6.1"],"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/concepts/co_63339ab48ab84253833955f9a8049f62"},{"public_id":"co_71bb5901b4bcf710a5868d30ccb2d55d","status":"active","name":"CellTiter-Blue","description":"An assay that measures resazurin reduction as an indicator of cell metabolic activity.","types":["assay"],"aliases":["CellTiter-Blue™"],"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/concepts/co_71bb5901b4bcf710a5868d30ccb2d55d"},{"public_id":"co_a5e71827d8aacc9f9ebec1fac967947a","status":"active","name":"Z′ value","description":"A statistical metric used to assess assay quality and separation between positive and negative controls.","types":["assay quality metric"],"aliases":["Z prime","Z-prime","Z′"],"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/concepts/co_a5e71827d8aacc9f9ebec1fac967947a"},{"public_id":"co_ab4256c81f5d92daa35f2f9981f78152","status":"active","name":"safety margins","description":"The separation between antibacterial activity and mammalian cell toxicity.","types":["screening criterion"],"aliases":["desired safety margins"],"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/concepts/co_ab4256c81f5d92daa35f2f9981f78152"},{"public_id":"co_ba59c4fb3e58e8816d51cba1efdd5646","status":"active","name":"antibacterial hits","description":"Compounds identified as active against bacteria in the screening campaign.","types":["compound set"],"aliases":["novel antibacterial hits"],"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/concepts/co_ba59c4fb3e58e8816d51cba1efdd5646"},{"public_id":"co_c046f9112705c273d79a6787c7343835","status":"active","name":"Toxilight","description":"An assay that uses adenylate kinase release as an indicator of membrane rupture.","types":["assay"],"aliases":["Toxilight™","adenylate kinase release assay"],"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/concepts/co_c046f9112705c273d79a6787c7343835"},{"public_id":"co_c86b1e0deaf371cb55dfeb1e8fc1d8c7","status":"active","name":"CellTiter-Glo","description":"An assay that measures intracellular adenosine triphosphate as an indicator of cell metabolic activity.","types":["assay"],"aliases":["CellTiter-Glo™"],"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/concepts/co_c86b1e0deaf371cb55dfeb1e8fc1d8c7"},{"public_id":"co_dc60c17e3b3a90cd729bf735f1b420b4","status":"active","name":"resazurin","description":"A redox dye reduced in the assay as a readout of metabolic activity.","types":["chemical","measurement reagent"],"aliases":[],"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/concepts/co_dc60c17e3b3a90cd729bf735f1b420b4"},{"public_id":"co_f33666cd9aea6356a07db03ff5a6ef41","status":"active","name":"intracellular adenosine triphosphate","description":"The intracellular energy molecule measured as a readout of cell metabolic activity.","types":["biomolecule","measurement readout"],"aliases":["ATP"],"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/concepts/co_f33666cd9aea6356a07db03ff5a6ef41"},{"public_id":"co_f5ba1e6429a6a71fecddca45a8a8048c","status":"active","name":"ChoK1","description":"A mammalian cell line used in the cytotoxicity assays.","types":["cell line"],"aliases":[],"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/concepts/co_f5ba1e6429a6a71fecddca45a8a8048c"}],"external_ids":{"DOI":"10.1177/1087057108329452","ArXiv":null,"PubMed":19196697,"PubMedCentral":null,"MAG":2125829020,"DBLP":null,"ACL":null},"open_access":{"is_open_access":false,"pdf_url":null,"landing_url":"https://sah.borca.ai/papers/23661802","source":null,"pdf_url_source":null,"license":null,"reason":"pdf_url_not_indexed"},"reference_availability":{"status":"available","references_indexed":true,"full_text_available":false,"full_text_source":null,"count_basis":"semantic_scholar_metadata","extraction_status":"not_applicable","reason":null},"source":{"provider":"episteme2","base_corpus":"semantic_scholar_dump","freshness_mode":"unknown","basis":["semantic_scholar_metadata","postgres_metadata"],"limits":["paper metadata is based on indexed upstream scholarly datasets","claims and concepts are available only for extracted papers","absence of claims or concepts means no extracted graph data is available in this response"],"status":"available","degraded":false,"degraded_reasons":[],"diagnostics":{"status":"available","degraded":false,"degraded_reasons":[],"metadata_status":"available","graph_status":"available","abstract_status":"available"},"source_flags":5},"paper_id":637148,"paper_uid":"9a2ab655-d52c-4ad1-a689-881cf0e412ac","canonical_identity":{"paper_id":637148,"paper_uid":"9a2ab655-d52c-4ad1-a689-881cf0e412ac","identity_status":"available","lookup_basis":"semantic_scholar_external_id","compatibility_path":"corpus_id"},"url":"https://sah.borca.ai/papers/23661802"}