{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,11]],"date-time":"2026-06-11T19:59:30Z","timestamp":1781207970528,"version":"3.54.1"},"reference-count":47,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2022,6,29]],"date-time":"2022-06-29T00:00:00Z","timestamp":1656460800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Glioblastoma (GBM) is an aggressive form of brain tumor with a median survival of approximately 12 months. With no new drugs in the last few decades and limited success in clinics for known therapies, drug repurposing is an attractive choice for its treatment. Here, we examined the efficacy of pyronaridine (PYR), an anti-malarial drug in GBM cells. PYR induced anti-proliferative activity in GBM cells with IC50 ranging from 1.16 to 6.82 \u00b5M. Synergistic activity was observed when PYR was combined with Doxorubicin and Ritonavir. Mechanistically, PYR triggered mitochondrial membrane depolarization and enhanced the ROS levels causing caspase-3 mediated apoptosis. PYR significantly decreased markers associated with proliferation, EMT, hypoxia, and stemness and upregulated the expression of E-cadherin. Interestingly, PYR induced the expression of intracellular as well as secretory Par-4, a tumor suppressor in GBM cells, which was confirmed using siRNA. Notably, Par-4 levels in plasma samples of GBM patients were significantly lower than normal healthy volunteers. Thus, our study demonstrates for the first time that PYR can be repurposed against GBM with a novel mechanism of action involving Par-4. Herewith, we discuss the role of upregulated Par-4 in a highly interconnected signaling network thereby advocating its importance as a therapeutic target.<\/jats:p>","DOI":"10.3390\/cancers14133198","type":"journal-article","created":{"date-parts":[[2022,6,29]],"date-time":"2022-06-29T20:47:56Z","timestamp":1656535676000},"page":"3198","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Prostate Apoptosis Response-4 (Par-4): A Novel Target in Pyronaridine-Induced Apoptosis in Glioblastoma (GBM) Cells"],"prefix":"10.3390","volume":"14","author":[{"given":"Jeevan","family":"Ghosalkar","sequence":"first","affiliation":[{"name":"Cell Biology Division, Cipla Ltd., Vikhroli, Mumbai 400083, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0128-5581","authenticated-orcid":false,"given":"Vinay","family":"Sonawane","sequence":"additional","affiliation":[{"name":"Cell Biology Division, Cipla Ltd., Vikhroli, Mumbai 400083, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tejal","family":"Pisal","sequence":"additional","affiliation":[{"name":"Cell Biology Division, Cipla Ltd., Vikhroli, Mumbai 400083, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5428-4358","authenticated-orcid":false,"given":"Swati","family":"Achrekar","sequence":"additional","affiliation":[{"name":"Cell Biology Division, Cipla Ltd., Vikhroli, Mumbai 400083, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Radha","family":"Pujari","sequence":"additional","affiliation":[{"name":"National Centre for Cell Sciences (NCCS), Pune 411007, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashish","family":"Chugh","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, D.Y. Patil Medical College, Pune 411018, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Padma","family":"Shastry","sequence":"additional","affiliation":[{"name":"National Centre for Cell Sciences (NCCS), Pune 411007, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1720-9519","authenticated-orcid":false,"given":"Kalpana","family":"Joshi","sequence":"additional","affiliation":[{"name":"Cell Biology Division, Cipla Ltd., Vikhroli, Mumbai 400083, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,6,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"987","DOI":"10.1056\/NEJMoa043330","article-title":"European Organisation for Research and Treatment of Cancer Brain Tumor and Radiotherapy Groups; National Cancer Institute of Canada Clinical Trials Group. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma","volume":"352","author":"Stupp","year":"2005","journal-title":"N. Engl. J. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"963","DOI":"10.3389\/fonc.2019.00963","article-title":"Glioblastoma Multiforme: An overview of emerging therapeutic targets","volume":"9","author":"Taylor","year":"2019","journal-title":"Front. Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"299","DOI":"10.3322\/caac.21613","article-title":"Management of glioblastoma: State of the art and future directions","volume":"70","author":"Tan","year":"2020","journal-title":"CA Cancer J. Clin."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"De Vleeschouwer, S. (2017). Epidemiology and Outcome of Glioblastoma. Glioblastoma, Codon Publications. Chapter 8.","DOI":"10.15586\/codon.glioblastoma.2017"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1073\/pnas.1913511116","article-title":"How tissue fluidity influences brain tumor progression","volume":"117","author":"Streitberger","year":"2020","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.ccr.2009.12.020","article-title":"Integrated genomic analysis identifies clinically relevant subtypes of glioblastoma characterized by abnormalities in PDGFRA, IDH1, EGFR, and NF1","volume":"17","author":"Verhaak","year":"2010","journal-title":"Cancer Cell"},{"key":"ref_7","first-page":"1","article-title":"Overcoming cancer therapeutic bottleneck by drug repurposing","volume":"5","author":"Zhang","year":"2020","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_8","first-page":"118","article-title":"Synthesis of a new antimalarial drug pyronaridine and its analogues","volume":"17","author":"Zheng","year":"1982","journal-title":"Acta Pharmacol. Sin."},{"key":"ref_9","first-page":"93","article-title":"In vitro activity of chloroquine, the two enantiomers of chloroquine, desethylchloroquine and pyronaridine against Plasmodium falciparum","volume":"22","author":"Fu","year":"1986","journal-title":"Br. J. Clin. Pharmacol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1186\/1475-2875-11-270","article-title":"Review of pyronaridine anti-malarial properties and product characteristics","volume":"11","author":"Croft","year":"2012","journal-title":"Malar J."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1016\/j.actatropica.2014.09.002","article-title":"Comparison of hematin-targeting properties of pynacrine, an acridine analog of the benzonaphthyridine antimalarial pyronaridine","volume":"140","author":"Sereekhajornjaru","year":"2014","journal-title":"Acta Tropica."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e23398","DOI":"10.1002\/bip.23398","article-title":"Pyronaridine: An update of its pharmacological activities and mechanisms of action","volume":"112","author":"Bailly","year":"2021","journal-title":"Biopolymers"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Villanueva, P.J., Martinez, A., Baca, S.T., DeJesus, R.E., Larragoity, M., Contreras, L., Gutierrez, D.A., Varela-Ramirez, A., and Aguilera, R.J. (2018). Pyronaridine exerts potent cytotoxicity on human breast and hematological cancer cells through induction of apoptosis. PLoS ONE., 13.","DOI":"10.1371\/journal.pone.0206467"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1111\/cbdd.13926","article-title":"Pyronaridine induces apoptosis in non-small cell lung cancer cells by upregulating death receptor 5 expression and inhibiting epidermal growth factor receptor","volume":"99","author":"Zhong","year":"2021","journal-title":"Chem. Biol. Drug Des."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"50","DOI":"10.2174\/2212697X08666210219101023","article-title":"The Antimalarial Drug Pyronaridine Inhibits Topoisomerase II in Breast Cancer Cells and Hinders Tumor Progression In Vivo","volume":"8","author":"Villanueva","year":"2021","journal-title":"Clin. Cancer Drugs"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"0652","DOI":"10.1089\/dna.2007.0652","article-title":"DNA Intercalation by Quinacrine and Methylene Blue: A Comparative Binding and thermodynamic characterization Study","volume":"27","author":"Hossain","year":"2008","journal-title":"DNA Cell Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1730","DOI":"10.1021\/bi201159b","article-title":"Amsacrine as a Topoisomerase II Poison: Importance of Drug\u2013DNA Interactions","volume":"51","author":"Ketron","year":"2012","journal-title":"Biochemistry"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"5045","DOI":"10.1038\/s41598-020-61876-5","article-title":"Piscidin-1 Induces Apoptosis via Mitochondrial Reactive Oxygen Species-Regulated Mitochondrial Dysfunction in Human Osteosarcoma Cells","volume":"10","author":"Cheng","year":"2020","journal-title":"Sci Rep."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1124","DOI":"10.1016\/j.bbrc.2004.05.099","article-title":"Pyronaridine, a novel modulator of P-glycoprotein-mediated multidrug resistance in tumor cells in vitro and in vivo","volume":"9","author":"Qi","year":"2004","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1785","DOI":"10.1016\/j.nano.2017.01.014","article-title":"pH, redox and photothermal tri-responsive DNA\/polyethylenimine conjugated gold nanorods as nanocarriers for specific intracellular co-release of Doxorubicin and chemosensitizer pyronaridine to combat multidrug resistant cancer","volume":"13","author":"Wang","year":"2017","journal-title":"Nanomedicine"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"508","DOI":"10.1016\/j.celrep.2016.12.051","article-title":"Chloroquine-inducible Par-4 secretion is essential for tumor cell apoptosis and inhibition of metastasis","volume":"18","author":"Burikhanov","year":"2017","journal-title":"Cell Rep."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3715","DOI":"10.1002\/jcp.24098","article-title":"Mechanisms of apoptosis by the tumor suppressor Par-4","volume":"227","author":"Hebbar","year":"2012","journal-title":"J. Cell. Physiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/j.toxlet.2013.10.008","article-title":"Par-4 downregulation confers cisplatin resistance in pancreatic cancer cells via PI3K\/Akt pathway-dependent EMT","volume":"224","author":"Tan","year":"2014","journal-title":"Toxicol Lett."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"3873","DOI":"10.1038\/onc.2010.141","article-title":"Cancer-selective apoptotic effects of extracellular and intracellular Par-4","volume":"29","author":"Rangnekar","year":"2010","journal-title":"Oncogene"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Ge, R., and Kao, C. (2019). Cell Surface GRP-78 as a Death Receptor and an Anticancer Drug Target. Cancers, 11.","DOI":"10.3390\/cancers11111787"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"6190","DOI":"10.1158\/0008-5472.CAN-08-0831","article-title":"Par-4 binds to topoisomerase 1 and attenuates its DNA relaxation activity","volume":"68","author":"Goswami","year":"2008","journal-title":"Cancer Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1011","DOI":"10.1158\/0008-5472.CAN-12-3212","article-title":"Novel Mechanism of Apoptosis Resistance in Cancer Mediated by Extracellular PAR-4","volume":"73","author":"Burikhanov","year":"2013","journal-title":"Can. Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1016\/j.cell.2009.07.007","article-title":"Cell death: A new Par-4 the TRAIL","volume":"138","author":"Hart","year":"2009","journal-title":"Cell"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Padovan, M., Eoli, M., Pellerino, A., Rizzato, S., Caserta, C., Simonelli, M., Michiara, M., Caccese, M., Anghileri, E., and Cerretti, G. (2021). Depatuxizumab Mafodotin (Depatux-M) Plus Temozolomide in Recurrent Glioblastoma Patients: Real-World Experience from a Multicenter Study of Italian Association of Neuro-Oncology (AINO). Cancers, 13.","DOI":"10.3390\/cancers13112773"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3139","DOI":"10.4161\/hv.29276","article-title":"DCVax\u00ae-L--developed by Northwest Biotherapeutics","volume":"10","author":"Polyzoidis","year":"2014","journal-title":"Hum. Vaccines Immunother."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"489","DOI":"10.4269\/ajtmh.2012.11-0558","article-title":"Drug-drug interaction analysis of pyronaridine\/artesunate and Ritonavir in healthy volunteers","volume":"86","author":"Morris","year":"2012","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2185","DOI":"10.1002\/ijc.26252","article-title":"ROS implication in a new antitumor strategy based on non-thermal plasma","volume":"130","author":"Vandamme","year":"2012","journal-title":"Int. J. Cancer."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1016\/j.ejca.2020.09.017","article-title":"Drug repositioning of antiretroviral Ritonavir for combinatorial therapy in glioblastoma","volume":"140","author":"Rauschenbach","year":"2020","journal-title":"Eup. J. Cancer"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1016\/j.gendis.2016.04.007","article-title":"Temozolomide resistance in glioblastoma multiforme","volume":"3","author":"Lee","year":"2016","journal-title":"Genes Dis."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"482","DOI":"10.1111\/j.1750-3639.2003.tb00479.x","article-title":"P-Glycoprotein and Multidrug Resistance-associated Protein Mediate Specific Patterns of Multidrug Resistance in Malignant Glioma Cell Lines, but not in Primary Glioma Cells","volume":"13","author":"Rieger","year":"2003","journal-title":"Brain Pathol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"20585","DOI":"10.1038\/srep20585","article-title":"Miltirone exhibits antileukemic activity by ROS-mediated endoplasmic reticulum stress and mitochondrial dysfunction pathways","volume":"6","author":"Zhou","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1007\/s00204-014-1249-z","article-title":"Aristolochic acid-induced apoptosis and G2 cell cycle arrest depends on ROS generation and MAP kinases activation","volume":"89","author":"Romanov","year":"2015","journal-title":"Arch. Toxicol."},{"key":"ref_38","first-page":"3825","article-title":"Local delivery of doxorubicin for the treatment of malignant brain tumors in rats","volume":"25","author":"Lesniak","year":"2005","journal-title":"Anticancer Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"11292","DOI":"10.1038\/s41598-020-68017-y","article-title":"Doxorubicin-loaded iron oxide nanoparticles for glioblastoma therapy: A combinational approach for enhanced delivery of nanoparticles","volume":"10","author":"Norouzi","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"4105","DOI":"10.2147\/IJN.S314895","article-title":"Doxorubicin Delivered via ApoE-Directed Reduction-Sensitive Polymersomes Potently Inhibit Orthotopic Human Glioblastoma Xenografts in Nude Mice","volume":"16","author":"Ouyang","year":"2021","journal-title":"Int. J. Nanomed."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Malfanti, A., Catania, G., Degros, Q., Wang, M., Bausart, M., and Pr\u00e9at, V. (2022). Design of Bio-Responsive Hyaluronic Acid-Doxorubicin Conjugates for the Local Treatment of Glioblastoma. Pharmaceutics, 14.","DOI":"10.3390\/pharmaceutics14010124"},{"key":"ref_42","first-page":"544","article-title":"Function and mechanism of pyronaridine: A new inhibitor of P-glycoprotein-mediated multidrug resistance","volume":"23","author":"Qi","year":"2002","journal-title":"Acta Pharmacol. Sin."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2496","DOI":"10.1159\/000495920","article-title":"Inhibition of Cyclin D1 Expression in Human Glioblastoma Cells is Associated with Increased Temozolomide Chemosensitivity","volume":"51","author":"Zhang","year":"2018","journal-title":"Cell Physiol. Biochem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1007\/s11060-011-0692-4","article-title":"Knockdown of cyclin D1 inhibits proliferation, induces apoptosis, and attenuates the invasive capacity of human glioblastoma cells","volume":"106","author":"Wang","year":"2012","journal-title":"J. Neurooncol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/j.fob.2014.11.005","article-title":"Secretory prostate apoptosis response (Par)-4 sensitizes multicellular spheroids (MCS) of glioblastoma multiforme cells to tamoxifen-induced cell death","volume":"5","author":"Jagtap","year":"2014","journal-title":"FEBS Open Bio."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1038\/s41419-020-03292-1","article-title":"Prostate apoptosis response-4 and tumor suppression: It\u2019s not just about apoptosis anymore","volume":"12","author":"Cheratta","year":"2021","journal-title":"Cell Death Dis."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1186\/s13287-021-02231-x","article-title":"Role of glioblastoma stem cells in cancer therapeutic resistance: A perspective on antineoplastic agents from natural sources and chemical derivatives","volume":"12","author":"Alves","year":"2021","journal-title":"Stem. Cell Res. Ther."}],"container-title":["Cancers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6694\/14\/13\/3198\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:40:42Z","timestamp":1760139642000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6694\/14\/13\/3198"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,6,29]]},"references-count":47,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["cancers14133198"],"URL":"https:\/\/doi.org\/10.3390\/cancers14133198","relation":{},"ISSN":["2072-6694"],"issn-type":[{"value":"2072-6694","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,6,29]]}}}