{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T10:06:06Z","timestamp":1785405966373,"version":"3.56.0"},"reference-count":32,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T00:00:00Z","timestamp":1785369600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Xinjiang Leading Talent Program: Science and Technology Innovation Platform Talent Attraction Project","award":["XJRC-2025-BTKJ-YJ-CXPT-229"],"award-info":[{"award-number":["XJRC-2025-BTKJ-YJ-CXPT-229"]}]},{"name":"Tarim University President Fund","award":["TDZKBS202633"],"award-info":[{"award-number":["TDZKBS202633"]}]},{"name":"Postgraduate Research and Innovation Program of Tarim University","award":["TDBSCX202504"],"award-info":[{"award-number":["TDBSCX202504"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>The development of dual-acting hybrid antibiotics is a promising strategy to combat the ongoing spread of drug-resistant tuberculosis. Inspired by the structure of the natural dithiolopyrrolone hybrid antibiotic thiomarinol and based on the synergistic effects confirmed by checkerboard assays, we employed a molecular hybridization strategy. The dithiolopyrrolone natural product thiolutin was covalently linked to the pharmacophores of four clinical antitubercular drugs\u2014cycloserine, linezolid, isoniazid, and pyrazinamide\u2014through alkyl linkers of 7\u201310 carbon atoms via amide condensation, leading to the design and synthesis of 15 novel hybrids. In vitro antitubercular activity evaluation revealed that the cycloserine series exhibited the best activity, with MIC values as low as 1 \u03bcg\/mL, followed by the isoniazid series. Cytotoxicity assays showed that all cycloserine hybrids had IC50 values &gt; 40 \u03bcg\/mL against RAW 264.7 mouse macrophages, markedly lower than that of thiolutin alone. Among them, T1-CS and T4-CS displayed the best selectivity indices, achieving an effective reduction in cytotoxicity. This study successfully constructed a class of thiolutin\u2013cycloserine hybrids with low cytotoxicity and high selectivity, providing a valuable molecular template for the discovery of novel antitubercular lead compounds.<\/jats:p>","DOI":"10.3390\/microorganisms14081670","type":"journal-article","created":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:32:13Z","timestamp":1785403933000},"page":"1670","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Design, Synthesis, and Antitubercular Activity of Thiolutin\u2013Cycloserine Hybrids: Reducing Cytotoxicity"],"prefix":"10.3390","volume":"14","author":[{"given":"Zhibin","family":"Sun","sequence":"first","affiliation":[{"name":"Key Laboratory of Conservation and Utilization of Biological Resources in the Tarim Basin, College of Life Science and Technology, Tarim University, Alaer 843300, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaolong","family":"Chen","sequence":"additional","affiliation":[{"name":"Key Laboratory of Conservation and Utilization of Biological Resources in the Tarim Basin, College of Life Science and Technology, Tarim University, Alaer 843300, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shanfeng","family":"Shi","sequence":"additional","affiliation":[{"name":"Key Laboratory of Conservation and Utilization of Biological Resources in the Tarim Basin, College of Life Science and Technology, Tarim University, Alaer 843300, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongqi","family":"Mu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Conservation and Utilization of Biological Resources in the Tarim Basin, College of Life Science and Technology, Tarim University, Alaer 843300, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chuanxing","family":"Wan","sequence":"additional","affiliation":[{"name":"Key Laboratory of Conservation and Utilization of Biological Resources in the Tarim Basin, College of Life Science and Technology, Tarim University, Alaer 843300, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9972-5225","authenticated-orcid":false,"given":"Guoguo","family":"He","sequence":"additional","affiliation":[{"name":"Key Laboratory of Conservation and Utilization of Biological Resources in the Tarim Basin, College of Life Science and Technology, Tarim University, Alaer 843300, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,30]]},"reference":[{"key":"ref_1","first-page":"1","article-title":"WHO 2025 global tuberculosis report 2025 key data analysis for the world and China","volume":"10","author":"He","year":"2025","journal-title":"Electron. J. Emerg. Infect. Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1115","DOI":"10.1080\/17460441.2025.2531229","article-title":"Understanding the key challenges in tuberculosis drug discovery: What does the future hold?","volume":"20","author":"Ramuz","year":"2025","journal-title":"Expert. Opin. Drug Discov."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Wu, R., Sun, M., Li, X., Fang, R., Xing, J., Li, Z., Wen, Y., and Song, N. (2025). Progress of anti-tuberculosis drug targets and novel therapeutic strategies. Front. Microbiol., 16.","DOI":"10.3389\/fmicb.2025.1637254"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1038\/s41579-020-0420-1","article-title":"Nanomaterial-based therapeutics for antibiotic-resistant bacterial infections","volume":"19","author":"Makabenta","year":"2021","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/j.cis.2017.07.030","article-title":"Natural and bioinspired nanostructured bactericidal surfaces","volume":"248","author":"Tripathy","year":"2017","journal-title":"Adv. Colloid. Interface Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"100851","DOI":"10.1016\/j.mtnano.2026.100851","article-title":"Geometry-driven mechanobactericidal activity on Escherichia coli and fibroblast biocompatibility of PMMA nanostructures fabricated by electron beam lithography","volume":"34","author":"Pellegrino","year":"2026","journal-title":"Mater. Today Nano"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2200608","DOI":"10.1002\/admi.202200608","article-title":"Mechano-Dynamic Analysis of the Bactericidal Activity of Bioinspired Moth-Eye Nanopatterned Surfaces","volume":"9","author":"Alameda","year":"2022","journal-title":"Adv. Mater. Interfaces"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"512","DOI":"10.1007\/s11094-025-03422-z","article-title":"Multi-Targeting Drugs to Overcome Multidrug-Resistant Tuberculosis: A Review","volume":"59","author":"Rao","year":"2025","journal-title":"Pharm. Chem. J."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1283","DOI":"10.1080\/17460441.2025.2552145","article-title":"The state of the art in dual-acting hybrid antibiotics to combat bacterial resistance","volume":"20","author":"Parkes","year":"2025","journal-title":"Expert. Opin. Drug Discov."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e2603100","DOI":"10.71150\/jm.2603100","article-title":"Pioneering strategies for overcoming bacterial drug resistance","volume":"64","author":"Kim","year":"2026","journal-title":"J. Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"582","DOI":"10.1021\/acsinfecdis.3c00504","article-title":"The Hybrid Antibiotic Thiomarinol A Overcomes Intrinsic Resistance in Escherichia coli Using a Privileged Dithiolopyrrolone Moiety","volume":"10","author":"Johnson","year":"2024","journal-title":"ACS Infect. Dis."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e202410799","DOI":"10.1002\/anie.202410799","article-title":"Bode, Isofunctional but Structurally Different Methyltransferases for Dithiolopyrrolone Diversification","volume":"63","author":"Su","year":"2024","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"3970","DOI":"10.3390\/md11103970","article-title":"Dithiolopyrrolone natural products: Isolation, synthesis and biosynthesis","volume":"11","author":"Qin","year":"2013","journal-title":"Mar. Drugs"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"905","DOI":"10.1039\/C3NP70106A","article-title":"Dithiolopyrrolones: Biosynthesis, synthesis, and activity of a unique class of disulfide-containing antibiotics","volume":"31","author":"Li","year":"2014","journal-title":"Nat. Prod. Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5137","DOI":"10.1002\/anie.201411667","article-title":"Enzymatic basis of \u201chybridity\u201d in thiomarinol biosynthesis","volume":"54","author":"Dunn","year":"2015","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1909","DOI":"10.1021\/acschembio.3c00360","article-title":"Chemoproteomics Reveals Disruption of Metal Homeostasis and Metalloproteins by the Antibiotic Holomycin","volume":"18","author":"Chan","year":"2023","journal-title":"ACS Chem. Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2717","DOI":"10.1073\/pnas.1612810114","article-title":"Role for dithiolopyrrolones in disrupting bacterial metal homeostasis","volume":"114","author":"Chan","year":"2017","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"12003","DOI":"10.1021\/jacs.1c02622","article-title":"Inhibition of Isoleucyl-tRNA Synthetase by the Hybrid Antibiotic Thiomarinol","volume":"143","author":"Johnson","year":"2021","journal-title":"J. Am. Chem. Soc."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e030239","DOI":"10.1101\/cshperspect.a030239","article-title":"Appropriate Targets for Antibacterial Drugs","volume":"6","author":"Silver","year":"2016","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref_20","first-page":"e67200","article-title":"Detection of Drug Susceptibility and Drug Interactions in Mycobacterium tuberculosis using Resazurin Microtiter and Checkerboard Assay","volume":"24","author":"Padmanaban","year":"2025","journal-title":"J. Vis. Exp."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"463","DOI":"10.7883\/yoken.66.463","article-title":"Determination of in vitro activities of polymyxin B and rifampin in combination with ampicillin\/sulbactam or cefoperazone\/sulbactam against multidrug-resistant Acinetobacter baumannii by the E-test and checkerboard methods","volume":"66","author":"Cetin","year":"2013","journal-title":"Jpn. J. Infect. Dis."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e14612","DOI":"10.1111\/cbdd.14612","article-title":"The Mycobacterium tuberculosis Cell Wall: An Alluring Drug Target for Developing Newer Anti-TB Drugs-A Perspective","volume":"104","author":"Chauhan","year":"2024","journal-title":"Chem. Biol. Drug Des."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1695","DOI":"10.1007\/s40265-022-01817-w","article-title":"The Struggle to End a Millennia-Long Pandemic: Novel Candidate and Repurposed Drugs for the Treatment of Tuberculosis","volume":"82","author":"Edwards","year":"2022","journal-title":"Drugs"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2546","DOI":"10.1093\/nar\/gkad1258","article-title":"Thiolutin has complex effects in vivo but is a direct inhibitor of RNA polymerase II in vitro","volume":"52","author":"Qiu","year":"2024","journal-title":"Nucleic Acids Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"662","DOI":"10.1038\/nm0696-662","article-title":"Mutations in pncA, a gene encoding pyrazinamidase\/nicotinamidase, cause resistance to the antituberculous drug pyrazinamide in tubercle bacillus","volume":"2","author":"Scorpio","year":"1996","journal-title":"Nat. Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"108500","DOI":"10.1016\/j.bioorg.2025.108500","article-title":"Design and synthesis of thiolutin derived PSMD14\/HDAC dual-target inhibitors against esophageal squamous cell carcinoma","volume":"161","author":"He","year":"2025","journal-title":"Bioorg. Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e0018125","DOI":"10.1128\/jb.00181-25","article-title":"Reductive activation of the disulfide-containing antibiotic thiolutin is mediated by both bacillithiol and FAD-dependent disulfide reductases","volume":"207","author":"Gaballa","year":"2025","journal-title":"J. Bacteriol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e202202567","DOI":"10.1002\/chem.202202567","article-title":"Dithiolopyrrolones are Prochelators that are Activated by Glutathione","volume":"29","author":"Albini","year":"2023","journal-title":"Chemistry"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1038\/nchembio.2370","article-title":"Thiolutin is a zinc chelator that inhibits the Rpn11 and other JAMM metalloproteases","volume":"13","author":"Lauinger","year":"2017","journal-title":"Nat. Chem. Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1983","DOI":"10.1042\/BCJ20200194","article-title":"The thick waxy coat of mycobacteria, a protective layer against antibiotics and the host\u2019s immune system","volume":"477","author":"Batt","year":"2020","journal-title":"Biochem. J."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1089\/ars.2017.7074","article-title":"Chemistry and redox biology of mycothiol","volume":"28","author":"Reyes","year":"2018","journal-title":"Antioxid. Redox Signal."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1128\/AAC.00558-10","article-title":"Structure of the Mycobacterium tuberculosis D-alanine:D-alanine ligase, a target of the antituberculosis drug D-cycloserine","volume":"55","author":"Bruning","year":"2011","journal-title":"Antimicrob. Agents Chemother."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/8\/1670\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:43:02Z","timestamp":1785404582000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/8\/1670"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,30]]},"references-count":32,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["microorganisms14081670"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms14081670","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,30]]}}}