{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T16:42:03Z","timestamp":1781628123729,"version":"3.54.5"},"reference-count":36,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2025,11,24]],"date-time":"2025-11-24T00:00:00Z","timestamp":1763942400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIH","award":["1R16GM149379"],"award-info":[{"award-number":["1R16GM149379"]}]},{"name":"Research Centers in Minority Institutions","award":["5U54MD007592"],"award-info":[{"award-number":["5U54MD007592"]}]},{"name":"NIGMS training","award":["T32GM144919"],"award-info":[{"award-number":["T32GM144919"]}]},{"name":"UTEP College of Science Dodson funds"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Histone acetylation and deacetylation are key regulators of gene expression and are frequently dysregulated in cancer, contributing to tumorigenesis and drug resistance. Overexpression of histone deacetylases (HDACs) in many cancer types leads to silencing of tumor suppressor genes and uncontrolled proliferation. Tumors often rely on epigenetic mechanisms to escape therapy and develop resistance. This study aimed to identify novel compounds that selectively target cancer cells while minimizing toxicity to non-cancerous cell lines. A series of novel HDAC inhibitors was evaluated using the Differential Nuclear Staining (DNS) assay, flow cytometry, and HDAC inhibition assays. These assays assessed cytotoxicity, selectivity, and mechanisms of cell death. Among seven compounds tested, VS-186B exhibited the highest cytotoxicity and Selective Cytotoxicity Index (SCI), particularly against the human Jurkat T-cell leukemia cell line. Flow cytometry experiments (Annexin V-FITC, ROS, JC-1, and Caspase-3\/7 assays) revealed that VS-186B induced apoptosis. VS-186B was more cytotoxic than Curcumin and Vorinostat across most of the cell lines tested and was more specific to hematological cells. Connectivity Map (CMap) analysis showed strong similarity to genes affected by known HDAC inhibitors. Subsequently, HDAC enzymatic assays confirmed that VS-186B inhibits Class I and II HDACs in a dose-dependent manner. VS-186B exhibits promising anticancer potential as a selective HDAC inhibitor since it induces apoptosis in cancer cells without significant cytotoxicity to non-cancerous lines with a similar gene expression profile to known HDAC inhibitors. These findings support further development of VS-186B as an epigenetic treatment for leukemia\/lymphoma.<\/jats:p>","DOI":"10.3390\/ijms262311354","type":"journal-article","created":{"date-parts":[[2025,11,24]],"date-time":"2025-11-24T14:59:04Z","timestamp":1763996344000},"page":"11354","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Functional Characterization of VS-186B, a Novel HDAC Inhibitor with Anticancer Activity"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5718-2181","authenticated-orcid":false,"given":"Laura A.","family":"Sanchez-Michael","sequence":"first","affiliation":[{"name":"Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX 79968, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-7024-5121","authenticated-orcid":false,"given":"Vijayalakshmi","family":"Sudarshan","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, KLE College of Pharmacy, Bengaluru 560010, Karnataka, India"},{"name":"KLE Academy of Higher Education and Research, KLE Deemed to be University, Belagavi 590010, Karnataka, India"},{"name":"Department of Pharmaceutical Chemistry, Faculty of Pharmacy, M.S. Ramaiah University of Applied Sciences, Bengaluru 560054, Karnataka, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-6605-459X","authenticated-orcid":false,"given":"Allison","family":"Elias","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX 79968, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5421-2242","authenticated-orcid":false,"given":"Denisse A.","family":"Gutierrez","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX 79968, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jose A.","family":"Lopez-Saenz","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX 79968, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6851-7231","authenticated-orcid":false,"given":"Jaqueline","family":"Pena-Zacarias","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX 79968, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-3054-3151","authenticated-orcid":false,"given":"Gabriela C.","family":"Torres","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX 79968, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2071-4874","authenticated-orcid":false,"given":"Armando","family":"Varela-Ramirez","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX 79968, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0833-158X","authenticated-orcid":false,"given":"Sujeet","family":"Kumar","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, NITTE College of Pharmaceutical Sciences, Nitte-Deemed to be University, Bengaluru 560064, Karnataka, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5599-3594","authenticated-orcid":false,"given":"Subhas S.","family":"Karki","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, KLE College of Pharmacy, Bengaluru 560010, Karnataka, India"},{"name":"KLE Academy of Higher Education and Research, KLE Deemed to be University, Belagavi 590010, Karnataka, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3765-2793","authenticated-orcid":false,"given":"Renato J.","family":"Aguilera","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX 79968, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Paro, P.D.R., Grossniklaus, P.D.U., Santoro, D.R., and Wutz, P.D.A. (2021). Epigenetics and Cancer. Introduction to Epigenetics [Internet], Springer.","DOI":"10.1007\/978-3-030-68670-3"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Jenke, R., Re\u00dfing, N., Hansen, F.K., Aigner, A., and B\u00fcch, T. (2021). Anticancer Therapy with HDAC Inhibitors: Mechanism-Based Combination Strategies and Future Perspectives. Cancers, 13.","DOI":"10.3390\/cancers13040634"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Morrison, O., and Thakur, J. (2021). Molecular Complexes at Euchromatin, Heterochromatin and Centromeric Chromatin. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22136922"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"a026831","DOI":"10.1101\/cshperspect.a026831","article-title":"HDACs and HDAC Inhibitors in Cancer Development and Therapy","volume":"6","author":"Li","year":"2016","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"117185","DOI":"10.1016\/j.ejmech.2024.117185","article-title":"Synthetic Approaches and Clinical Applications of Representative HDAC Inhibitors for Cancer Therapy: A Review","volume":"283","author":"Lv","year":"2025","journal-title":"Eur. J. Med. Chem."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Karagiannis, D., and Rampias, T. (2021). HDAC Inhibitors: Dissecting Mechanisms of Action to Counter Tumor Heterogeneity. Cancers, 13.","DOI":"10.3390\/cancers13143575"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"117022","DOI":"10.1016\/j.ejmech.2024.117022","article-title":"Tubulin\/HDAC Dual-Target Inhibitors: Insights from Design Strategies, SARs, and Therapeutic Potential","volume":"281","author":"Zhang","year":"2025","journal-title":"Eur. J. Med. Chem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1038\/s44276-024-00064-8","article-title":"New Light on Chemotherapy Toxicity and Its Prevention","volume":"2","author":"Juthani","year":"2024","journal-title":"BJC Rep."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"339","DOI":"10.15171\/apb.2017.041","article-title":"The Different Mechanisms of Cancer Drug Resistance: A Brief Review","volume":"7","author":"Mansoori","year":"2017","journal-title":"Adv. Pharm. Bull."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1367","DOI":"10.1016\/j.gendis.2022.02.007","article-title":"Cancer Chemotherapy and beyond: Current Status, Drug Candidates, Associated Risks and Progress in Targeted Therapeutics","volume":"10","author":"Anand","year":"2023","journal-title":"Genes Dis."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Saeed, M.E.M., Y\u00fccer, R., Dawood, M., Hegazy, M.-E.F., Drif, A., Ooko, E., Kadioglu, O., Seo, E.-J., Kamounah, F.S., and Titinchi, S.J. (2022). In Silico and In Vitro Screening of 50 Curcumin Compounds as EGFR and NF-\u03baB Inhibitors. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23073966"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Mazumder, K., Aktar, A., Roy, P., Biswas, B., Hossain, M.E., Sarkar, K.K., Bachar, S.C., Ahmed, F., Monjur-Al-Hossain, A.S.M., and Fukase, K. (2022). A Review on Mechanistic Insight of Plant Derived Anticancer Bioactive Phytocompounds and Their Structure Activity Relationship. Molecules, 27.","DOI":"10.3390\/molecules27093036"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1021\/acsptsci.2c00012","article-title":"Role of Turmeric and Curcumin in Prevention and Treatment of Chronic Diseases: Lessons Learned from Clinical Trials","volume":"6","author":"Kunnumakkara","year":"2023","journal-title":"ACS Pharmacol. Transl. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"7035","DOI":"10.3390\/molecules15107035","article-title":"Potential of the Dietary Antioxidants Resveratrol and Curcumin in Prevention and Treatment of Hematologic Malignancies","volume":"15","author":"Kelkel","year":"2010","journal-title":"Molecules"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1234","DOI":"10.1016\/j.fct.2010.02.015","article-title":"Curcumin Supplementation Protects from Genotoxic Effects of Arsenic and Fluoride","volume":"48","author":"Tiwari","year":"2010","journal-title":"Food Chem. Toxicol."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Allegra, A., Mirabile, G., Ettari, R., Pioggia, G., and Gangemi, S. (2022). The Impact of Curcumin on Immune Response: An Immunomodulatory Strategy to Treat Sepsis. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232314710"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Razali, N.S.C., Lam, K.W., Rajab, N.F., Jamal, A.R.A., Kamaluddin, N.F., and Chan, K.M. (2022). Curcumin Piperidone Derivatives Induce Anti-Proliferative and Anti-Migratory Effects in LN-18 Human Glioblastoma Cells. Sci. Rep., 12.","DOI":"10.1038\/s41598-022-16274-4"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Dei Cas, M., and Ghidoni, R. (2019). Dietary Curcumin: Correlation between Bioavailability and Health Potential. Nutrients, 11.","DOI":"10.3390\/nu11092147"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"102082","DOI":"10.1016\/j.jddst.2020.102082","article-title":"Recent Developments in Formulation Design for Improving Oral Bioavailability of Curcumin: A Review","volume":"60","author":"Liu","year":"2020","journal-title":"J. Drug Deliv. Sci. Technol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"7449","DOI":"10.1021\/bi400734e","article-title":"Stable and Potent Analogues Derived from the Modification of the Dicarbonyl Moiety of Curcumin","volume":"52","author":"Chakraborti","year":"2013","journal-title":"Biochemistry"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Kaur, K., Al-Khazaleh, A.K., Bhuyan, D.J., Li, F., and Li, C.G. (2024). A Review of Recent Curcumin Analogues and Their Antioxidant, Anti-Inflammatory, and Anticancer Activities. Antioxidants, 13.","DOI":"10.20944\/preprints202403.1292.v1"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"5593","DOI":"10.1007\/s11696-025-04161-x","article-title":"Monocarbonyl Modifications of Curcumin: An Overview of Structure\u2013Activity Relationship Study as Anti-Infective Agents","volume":"79","author":"Teli","year":"2025","journal-title":"Chem. Pap."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"102334","DOI":"10.1016\/j.jddst.2021.102334","article-title":"Delivery Systems for Vorinostat in Cancer Treatment: An Updated Review","volume":"61","author":"Le","year":"2021","journal-title":"J. Drug Deliv. Sci. Technol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Squarzoni, A., Scuteri, A., and Cavaletti, G. (2022). HDACi: The Columbus\u2019 Egg in Improving Cancer Treatment and Reducing Neurotoxicity?. Cancers, 14.","DOI":"10.3390\/cancers14215251"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1007\/s13402-016-0272-x","article-title":"A Novel Curcumin-like Dienone Induces Apoptosis in Triple-Negative Breast Cancer Cells","volume":"39","author":"Das","year":"2016","journal-title":"Cell. Oncol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"655","DOI":"10.1021\/acs.jmedchem.5b01525","article-title":"Design, Synthesis, and Pharmacological Evaluation of Novel N-Acylhydrazone Derivatives as Potent Histone Deacetylase 6\/8 Dual Inhibitors","volume":"59","author":"Rodrigues","year":"2016","journal-title":"J. Med. Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1038\/icb.2011.100","article-title":"HDAC Inhibitors in Cancer Biology: Emerging Mechanisms and Clinical Applications","volume":"90","author":"Khan","year":"2012","journal-title":"Immunol. Cell Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1038\/nrc1893","article-title":"Drug Penetration in Solid Tumours","volume":"6","author":"Minchinton","year":"2006","journal-title":"Nat. Rev. Cancer"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Saggar, J.K., Yu, M., Tan, Q., and Tannock, I.F. (2013). The Tumor Microenvironment and Strategies to Improve Drug Distribution. Front. Oncol., 3.","DOI":"10.3389\/fonc.2013.00154"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Choi, I.-K., Strauss, R., Richter, M., Yun, C.-O., and Lieber, A. (2013). Strategies to Increase Drug Penetration in Solid Tumors. Front. Oncol., 3.","DOI":"10.3389\/fonc.2013.00193"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Simsek, H., and Klotzsch, E. (2022). The Solid Tumor Microenvironment\u2014Breaking the Barrier for T Cells. BioEssays, 44.","DOI":"10.1002\/bies.202100285"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3487","DOI":"10.1111\/cas.15890","article-title":"Multifaced Roles of Desmoplastic Reaction and Fibrosis in Pancreatic Cancer Progression: Current Understanding and Future Directions","volume":"114","author":"Sato","year":"2023","journal-title":"Cancer Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"231","DOI":"10.4161\/cam.20419","article-title":"Understanding the Role of Stromal Fibroblasts in Cancer Progression","volume":"6","author":"Tripathi","year":"2012","journal-title":"Cell Adhes. Migr."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Xu, Z., Zhang, Y., Dai, H., and Han, B. (2022). Epithelial\u2013Mesenchymal Transition-Mediated Tumor Therapeutic Resistance. Molecules, 27.","DOI":"10.3390\/molecules27154750"},{"key":"ref_35","first-page":"1","article-title":"Differential Nuclear Staining Assay for High-Throughput Screening to Identify Cytotoxic Compounds","volume":"1","author":"Lema","year":"2011","journal-title":"Curr. Cell. Biochem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1007\/s10565-010-9178-y","article-title":"Cytotoxic Effects of Two Organotin Compounds and Their Mode of Inflicting Cell Death on Four Mammalian Cancer Cells","volume":"27","author":"Costanzo","year":"2011","journal-title":"Cell Biol. Toxicol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/23\/11354\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,27]],"date-time":"2025-11-27T05:14:06Z","timestamp":1764220446000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/23\/11354"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,24]]},"references-count":36,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["ijms262311354"],"URL":"https:\/\/doi.org\/10.3390\/ijms262311354","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,24]]}}}