{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T17:42:35Z","timestamp":1792086155461,"version":"4.4.0"},"reference-count":69,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"44","content-domain":{"domain":["www.science.org"],"crossmark-restriction":true},"short-container-title":["Sci. Adv."],"published-print":{"date-parts":[[2025,10,31]]},"abstract":"<jats:p>Covalent inhibitors that are approved and marketed drugs exploit a wide array of warheads and reactions with amino acid side chain\u2013based nucleophiles. Thiostrepton (TS) inhibits the peroxidase activity of the mitochondrial antioxidant protein peroxiredoxin 3 by forming a covalent crosslink between the two active site cysteine residues. Peroxiredoxin 3 inactivation increases reactive oxygen species levels, induces cancer cell death in preclinical models, and shows promise in an ongoing clinical trial for malignant mesothelioma using direct pleural infusion. We report the identification of the minimal fragment of TS that contains tandem dehydro-alanine (DHA) moieties and maintains anticancer activity while losing interactions with three alternative targets of intact TS. Biochemical, kinetic, cellular, and structural studies demonstrate that this fragment is a mechanism-based peroxiredoxin inhibitor. These findings represent a promising start toward a pro-oxidant approach for cancer therapy. Moreover, the data support that the DHA moiety should be added to the covalent warhead arsenal.<\/jats:p>","DOI":"10.1126\/sciadv.ady4492","type":"journal-article","created":{"date-parts":[[2025,10,31]],"date-time":"2025-10-31T17:58:57Z","timestamp":1761933537000},"update-policy":"https:\/\/doi.org\/10.34133\/aaas_crossmark","source":"Crossref","is-referenced-by-count":7,"title":["Mechanism-based peroxiredoxin 3 inhibitors exploit a covalent warhead for cancer therapy"],"prefix":"10.1126","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2464-7696","authenticated-orcid":true,"given":"Kimberly J.","family":"Nelson","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Center for Structural Biology, Wake Forest University School of Medicine, Medical Center Blvd., Winston-Salem, NC 27157, USA."},{"name":"Atrium Health Wake Forest Baptist Comprehensive Cancer Center, Medical Center Blvd., Winston-Salem, NC 27157, USA."},{"name":"Center for Redox Biology and Medicine, Wake Forest University School of Medicine, Medical Center Blvd., Winston-Salem, NC 27157, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5809-5864","authenticated-orcid":true,"suffix":"Jr.","given":"Terrence L.","family":"Smalley","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Center for Structural Biology, Wake Forest University School of Medicine, Medical Center Blvd., Winston-Salem, NC 27157, USA."},{"name":"Atrium Health Wake Forest Baptist Comprehensive Cancer Center, Medical Center Blvd., Winston-Salem, NC 27157, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4381-5694","authenticated-orcid":true,"given":"Terri","family":"Messier","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, University of Vermont Cancer Center, Larner College of Medicine, Burlington, VT 05405, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rajesh","family":"Gumpena","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Center for Structural Biology, Wake Forest University School of Medicine, Medical Center Blvd., Winston-Salem, NC 27157, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Uma","family":"Gandhi","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Center for Structural Biology, Wake Forest University School of Medicine, Medical Center Blvd., Winston-Salem, NC 27157, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0933-9065","authenticated-orcid":true,"given":"Stephanie","family":"Milczarek","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, University of Vermont Cancer Center, Larner College of Medicine, Burlington, VT 05405, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aida","family":"Habibovic","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, University of Vermont Cancer Center, Larner College of Medicine, Burlington, VT 05405, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hattie","family":"Hoffman","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Center for Structural Biology, Wake Forest University School of Medicine, Medical Center Blvd., Winston-Salem, NC 27157, USA."},{"name":"Wake Forest University, Biochemistry and Molecular Biology Program, Winston-Salem, NC 27109, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victoria","family":"Gibson","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, University of Vermont Cancer Center, Larner College of Medicine, Burlington, VT 05405, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1466-2852","authenticated-orcid":true,"given":"Robert J.","family":"Hondal","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Vermont, Burlington, VT, 05405, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0246-1295","authenticated-orcid":true,"given":"W. Todd","family":"Lowther","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Center for Structural Biology, Wake Forest University School of Medicine, Medical Center Blvd., Winston-Salem, NC 27157, USA."},{"name":"Atrium Health Wake Forest Baptist Comprehensive Cancer Center, Medical Center Blvd., Winston-Salem, NC 27157, USA."},{"name":"Center for Redox Biology and Medicine, Wake Forest University School of Medicine, Medical Center Blvd., Winston-Salem, NC 27157, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6035-2134","authenticated-orcid":true,"given":"Brian","family":"Cunniff","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, University of Vermont Cancer Center, Larner College of Medicine, Burlington, VT 05405, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1039\/D0MD00154F"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41573-022-00542-z"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1080\/13543776.2024.2400175"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jmedchem.3c01825"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1042\/BJ20091541"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkz430"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2007.02.022"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.3390\/antiox9020182"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0127310"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.3892\/mmr.2013.1370"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.3390\/antiox10020150"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/s43018-019-0018-6"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1200\/JCO.2024.42.16_suppl.TPS8124"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1200\/JCO.2024.42.16_suppl.3019"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1038\/nchem.1114"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0006593"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2005.09.103"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2008.01.009"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1002\/anie.200906988"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.7b08775"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1039\/C4QO00288A"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.RA117.001690"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M113.473470"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja0552803"},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja052934z"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature06415"},{"key":"e_1_3_2_28_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M109.036400"},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.9b02822"},{"key":"e_1_3_2_30_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi0517887"},{"key":"e_1_3_2_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2013.08.013"},{"key":"e_1_3_2_32_2","doi-asserted-by":"publisher","DOI":"10.1111\/jcmm.12602"},{"key":"e_1_3_2_33_2","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms6165"},{"key":"e_1_3_2_34_2","unstructured":"USP Antibiotics-Microbial Assays <81> in USP37-NF32 (USP 2014) p. 77\u201392."},{"key":"e_1_3_2_35_2","doi-asserted-by":"publisher","DOI":"10.3390\/antiox8010011"},{"key":"e_1_3_2_36_2","doi-asserted-by":"publisher","DOI":"10.3390\/antiox10020302"},{"key":"e_1_3_2_37_2","doi-asserted-by":"publisher","DOI":"10.3390\/biology12050666"},{"key":"e_1_3_2_38_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.redox.2014.11.003"},{"key":"e_1_3_2_39_2","doi-asserted-by":"publisher","DOI":"10.1021\/acschembio.0c00364"},{"key":"e_1_3_2_40_2","doi-asserted-by":"publisher","DOI":"10.1002\/chem.201803144"},{"key":"e_1_3_2_41_2","doi-asserted-by":"publisher","DOI":"10.1002\/chem.201902907"},{"key":"e_1_3_2_42_2","doi-asserted-by":"publisher","DOI":"10.1039\/D3SC02818F"},{"key":"e_1_3_2_43_2","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201911886"},{"key":"e_1_3_2_44_2","doi-asserted-by":"publisher","DOI":"10.1021\/tx100413v"},{"key":"e_1_3_2_45_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi801718d"},{"key":"e_1_3_2_46_2","doi-asserted-by":"publisher","DOI":"10.3390\/antiox9121220"},{"key":"e_1_3_2_47_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2013.06.011"},{"key":"e_1_3_2_48_2","doi-asserted-by":"publisher","DOI":"10.1039\/b901735f"},{"key":"e_1_3_2_49_2","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.935"},{"key":"e_1_3_2_50_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2019.04.003"},{"key":"e_1_3_2_51_2","doi-asserted-by":"publisher","DOI":"10.18632\/oncotarget.6435"},{"key":"e_1_3_2_52_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2018.11.013"},{"key":"e_1_3_2_53_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41392-022-01231-4"},{"key":"e_1_3_2_54_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2023.02.003"},{"key":"e_1_3_2_55_2","doi-asserted-by":"publisher","DOI":"10.1089\/ars.2010.3624"},{"key":"e_1_3_2_56_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jpha.2022.06.001"},{"key":"e_1_3_2_57_2","doi-asserted-by":"publisher","DOI":"10.3390\/molecules20034928"},{"key":"e_1_3_2_58_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jmedchem.9b00598"},{"key":"e_1_3_2_59_2","doi-asserted-by":"publisher","DOI":"10.18632\/oncotarget.6763"},{"key":"e_1_3_2_60_2","doi-asserted-by":"publisher","DOI":"10.1089\/ars.2016.6703"},{"key":"e_1_3_2_61_2","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1700796"},{"key":"e_1_3_2_62_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi200935d"},{"key":"e_1_3_2_63_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0703774104"},{"key":"e_1_3_2_64_2","doi-asserted-by":"publisher","DOI":"10.1089\/ars.2016.6922"},{"key":"e_1_3_2_65_2","doi-asserted-by":"publisher","DOI":"10.3390\/antiox10060946"},{"key":"e_1_3_2_66_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444996012255"},{"key":"e_1_3_2_67_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444904019158"},{"key":"e_1_3_2_68_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0021889892009944"},{"key":"e_1_3_2_69_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444909042073"},{"key":"e_1_3_2_70_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0039404"}],"container-title":["Science Advances"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/sciadv.ady4492","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,31]],"date-time":"2025-10-31T17:59:37Z","timestamp":1761933577000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/sciadv.ady4492"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,31]]},"references-count":69,"journal-issue":{"issue":"44","published-print":{"date-parts":[[2025,10,31]]}},"alternative-id":["10.1126\/sciadv.ady4492"],"URL":"https:\/\/doi.org\/10.1126\/sciadv.ady4492","relation":{},"ISSN":["2375-2548"],"issn-type":[{"value":"2375-2548","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,31]]},"assertion":[{"value":"2025-04-22","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-10-03","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-10-31","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"eady4492"}}