{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T06:53:13Z","timestamp":1790232793798,"version":"4.1.0"},"reference-count":50,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2019,10,22]],"date-time":"2019-10-22T00:00:00Z","timestamp":1571702400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Non-Small Cell Lung Cancer (NSCLC) is a common malignancy and leading cause of death by cancer. Metastasis and drug resistance are serious clinical problems encountered in NSCLC therapy. Aberrant activation of the Transforming Growth Factor beta (TGF\u03b2) and Hedgehog (Hh) signal transduction cascades often associate with poor prognosis and aggressive disease progression in NSCLC, as these signals can drive cell proliferation, angiogenesis, metastasis, immune evasion and emergence of drug resistance. Therefore, simultaneous inhibition of TGF\u03b2 and Hh signaling, by a single agent, or in combination with other drugs, could yield therapeutic benefits in NSCLC and other cancers. In the current study, we report on the biological and pharmacological evaluation of Oxy210, an oxysterol-based dual inhibitor of TGF\u03b2 and Hh signaling. In NSCLC cells, Oxy210 inhibits proliferation, epithelial-mesenchymal transition (EMT) and invasive activity. Combining Oxy210 with Carboplatin (CP) increases the anti-proliferative response to CP and inhibits TGF\u03b2-induced resistance to CP in A549 NSCLC cells. In addition, Oxy210 displays encouraging drug-like properties, including chemical scalability, metabolic stability and oral bioavailability in mice. Unlike other known inhibitors, Oxy210 antagonizes TGF\u03b2 and Hh signaling independently of TGF\u03b2 receptor kinase inhibition and downstream of Smoothened, respectively.<\/jats:p>","DOI":"10.3390\/cells8101297","type":"journal-article","created":{"date-parts":[[2019,10,23]],"date-time":"2019-10-23T11:46:59Z","timestamp":1571831219000},"page":"1297","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Inhibition of Non-Small Cell Lung Cancer Cells by Oxy210, an Oxysterol-Derivative that Antagonizes TGF\u03b2 and Hedgehog Signaling"],"prefix":"10.3390","volume":"8","author":[{"given":"Frank","family":"Stappenbeck","sequence":"first","affiliation":[{"name":"MAX BioPharma, Inc., 2870 Colorado Avenue, Santa Monica, CA 90404, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Feng","family":"Wang","sequence":"additional","affiliation":[{"name":"MAX BioPharma, Inc., 2870 Colorado Avenue, Santa Monica, CA 90404, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liu-Ya","family":"Tang","sequence":"additional","affiliation":[{"name":"Laboratory of Cellular &amp; Molecular Biology, Center for Cancer Research, National Cancer Institute, 37 Convent Drive, 37\/2056B Bethesda, MD 20892-4256, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2753-7601","authenticated-orcid":false,"given":"Ying E.","family":"Zhang","sequence":"additional","affiliation":[{"name":"Laboratory of Cellular &amp; Molecular Biology, Center for Cancer Research, National Cancer Institute, 37 Convent Drive, 37\/2056B Bethesda, MD 20892-4256, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Farhad","family":"Parhami","sequence":"additional","affiliation":[{"name":"MAX BioPharma, Inc., 2870 Colorado Avenue, Santa Monica, CA 90404, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,10,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"288","DOI":"10.21037\/tlcr.2016.06.07","article-title":"Non-small cell lung cancer: Current treatment and future advances","volume":"5","author":"Zappa","year":"2016","journal-title":"Transl. Lung Cancer Res."},{"key":"ref_2","unstructured":"American Cancer Society (2018). Cancer facts and figures. ACS, 18."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1016\/j.ctrv.2013.10.001","article-title":"The role of the tumor-microenvironment in lung cancer-metastasis and its relationship to potential therapeutic targets","volume":"40","author":"Wood","year":"2014","journal-title":"Cancer Treat. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1864","DOI":"10.1056\/NEJMra1504688","article-title":"Clinical implications of genomic discoveries in lung cancer","volume":"374","author":"Swanton","year":"2016","journal-title":"New Engl. J. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2647","DOI":"10.1159\/000495956","article-title":"Loss of Tumor Suppressor Gene Function in Human Cancer: An Overview","volume":"51","author":"Wang","year":"2018","journal-title":"Cell Physiol. Biochem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1016\/j.trecan.2016.05.010","article-title":"Resisting Resistance: Targeted Therapies in Lung Cancer","volume":"2","author":"Lin","year":"2016","journal-title":"Trends Cancer"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"653","DOI":"10.1586\/14737140.2016.1170596","article-title":"Platinum-based chemotherapy in advanced non-small-cell lung cancer: Optimal number of treatment cycles","volume":"16","author":"Rossi","year":"2016","journal-title":"Expert Rev. Anticancer Ther."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/j.ctrv.2017.10.002","article-title":"PD-1 blockade in advanced NSCLC: A focus on pembrolizumab","volume":"62","author":"Peters","year":"2018","journal-title":"Cancer Treat. Rev."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1038\/nrclinonc.2015.61","article-title":"Targeting Notch, Hedgehog, and Wnt pathways in cancer stem cells: Clinical update","volume":"12","author":"Takebe","year":"2015","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/j.pharmthera.2012.10.002","article-title":"Overlapping activities of TGF-\u03b2 and Hedgehog signaling in cancer: Therapeutic targets for cancer treatment","volume":"137","author":"Perrot","year":"2013","journal-title":"Pharmacol. Ther."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"754","DOI":"10.1378\/chest.125.2.754","article-title":"The role of transforming growth factor beta in lung development and disease","volume":"125","author":"Bartram","year":"2004","journal-title":"Chest"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1165\/rcmb.2014-0132TR","article-title":"Sonic hedgehog signaling in the lung. From development to disease","volume":"52","author":"Kugler","year":"2015","journal-title":"Am. J. Respir. Cell Mol. Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"556","DOI":"10.3389\/fgene.2019.00556","article-title":"Non-canonical Hedgehog Signaling Pathway in Cancer: Activation of GLI Transcription Factors Beyond Smoothened","volume":"10","author":"Pietrobono","year":"2019","journal-title":"Front Genet."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"a022129","DOI":"10.1101\/cshperspect.a022129","article-title":"Non-Smad Signaling Pathways of the TGF-\u03b2 Family","volume":"9","author":"Zhang","year":"2017","journal-title":"Cold Spring Harb. Perspect. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"15495","DOI":"10.1074\/jbc.M113.545194","article-title":"The transcription factor GLI1 interacts with SMAD proteins to modulate transforming growth factor \u03b2-induced gene expression in a p300\/CREB-binding protein-associated factor (PCAF)-dependent manner","volume":"289","author":"Nye","year":"2014","journal-title":"J. Biol. Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"022137","DOI":"10.1101\/cshperspect.a022137","article-title":"Signaling Cross Talk between TGF-\u03b2\/Smad and Other Signaling Pathways","volume":"9","author":"Luo","year":"2017","journal-title":"Cold Spring Harb. Perspect. Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1186\/1477-7819-12-253","article-title":"Expression of Gli1 family genes is associated with tumor progression in advanced lung adenocarcinoma","volume":"12","author":"Ishikawa","year":"2014","journal-title":"World J. Surg. Oncol."},{"key":"ref_18","first-page":"9766","article-title":"The relationship of transforming growth factor-\u03b2 and lung cancer","volume":"9","author":"Xue","year":"2016","journal-title":"Int. J. Clin. Exp. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1186\/s12943-016-0509-3","article-title":"Hedgehog signaling: Modulation of cancer properties and tumor mircroenvironment","volume":"15","author":"Hanna","year":"2016","journal-title":"Mol. Cancer"},{"key":"ref_20","first-page":"9732939","article-title":"Transforming of the Tumor Microenvironment: Implications for TGF-\u03b2 Inhibition in the Context of Immune-Checkpoint Therapy","volume":"2018","year":"2018","journal-title":"J. Oncol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Olkkonen, V.M., B\u00e9aslas, O., and Nissil\u00e4, E. (2012). Oxysterols and their cellular effectors. Biomolecules, 2.","DOI":"10.3390\/biom2010076"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1002\/jcb.25668","article-title":"Inhibition of Pancreatic Cancer Cell-Induced Paracrine Hedgehog Signaling by Liver X Receptor Agonists and Oxy16, a Naturally Occurring Oxysterol","volume":"118","author":"Wang","year":"2017","journal-title":"J. Cell. Biochem."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Wang, F., Stappenbeck, F., and Parhami, F. (2019). Inhibition of Hedgehog Signaling in Fibroblasts, Pancreatic, and Lung Tumor Cells by Oxy186, an Oxysterol Analogue with Drug-Like Properties. Cells, 8.","DOI":"10.3390\/cells8050509"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1016\/j.steroids.2004.04.005","article-title":"Stereoselective synthesis of (22R)- and (22S)-castasterone\/ponasterone A hybrid compounds and evaluation of their molting hormone activity","volume":"69","author":"Watanabe","year":"2004","journal-title":"Steroids"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"8959","DOI":"10.1074\/jbc.M611741200","article-title":"Oxysterols are novel activators of the hedgehog signaling pathway in pluripotent mesenchymal cells","volume":"282","author":"Dwyer","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1131","DOI":"10.1002\/jcb.21112","article-title":"Oxysterol-induced osteoblastic differentiation of pluripotent mesenchymal cells is mediated through a PKC- and PKA dependent pathway","volume":"100","author":"Richardson","year":"2007","journal-title":"J. Cell. Biochem."},{"key":"ref_27","first-page":"3810","article-title":"Type I receptors specify growth-inhibitory and transcriptional responses to transforming growth factor beta and activin","volume":"14","author":"Weis","year":"1994","journal-title":"Mol. Cell. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"918","DOI":"10.1016\/j.molcel.2008.09.002","article-title":"TRAF6 mediates Smad-independent activation of JNK and p38 by TGF-beta","volume":"31","author":"Yamashita","year":"2008","journal-title":"Mol. Cell."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2199","DOI":"10.1002\/j.1460-2075.1995.tb07214.x","article-title":"GS domain mutations that constitutively activate T beta R-I, the downstream signaling component in the TGF-beta receptor complex","volume":"14","author":"Wieser","year":"1995","journal-title":"EMBO J."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1038\/ng916","article-title":"Mutations in SUFU predispose to medulloblastoma","volume":"31","author":"Taylor","year":"2002","journal-title":"Nat. Genet."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Xie, H., Paradise, B.D., Ma, W.W., and Fernandez-Zapico, M.E. (2019). Recent Advances in the Clinical Targeting of Hedgehog\/GLI Signaling in Cancer. Cells, 8.","DOI":"10.3390\/cells8050394"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Norsworthy, K.J., By, K., Subramaniam, S., Zhuang, L., Del Valle, P.L., Przepiorka, D., Shen, Y.L., Sheth, C.M., Liu, C., and Leong, R. (2019). FDA Approval Summary: Glasdegib for newly-diagnosed acute myeloid leukemia. Clin. Cancer Res., 365.","DOI":"10.1158\/1078-0432.CCR-19-0365"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"7969286","DOI":"10.1155\/2016\/7969286","article-title":"The hedgehog signaling networks in lung cancer: The mechanisms and roles in tumor progression and implications for cancer therapy","volume":"2016","author":"Abe","year":"2016","journal-title":"Biomed. Res. Int."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Giroux-Leprieur, E., Costantini, A., Ding, V.W., and He, B. (2018). Hedgehog signaling in lung cancer: From oncogenesis to cancer treatment resistance. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19092835"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1016\/j.pharmthera.2017.11.004","article-title":"TGF\u03b2 pathway inhibition in the treatment of non-small cell lung cancer","volume":"184","author":"Eser","year":"2018","journal-title":"Pharmacol. Ther."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1844","DOI":"10.3892\/ijo.2015.2861","article-title":"Critical contribution of MCL-1 in EMT-associated chemo-resistance in A549 non-small cell lung cancer","volume":"46","author":"Toge","year":"2015","journal-title":"Int. J. Oncol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"766","DOI":"10.1093\/carcin\/bgx063","article-title":"Matrix metalloproteinases: Their functional role in lung cancer","volume":"38","author":"Merchant","year":"2017","journal-title":"Carcinogenesis"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ceb.2017.10.001","article-title":"Mechanistic insight into contextual TGF-\u03b2 signaling","volume":"51","author":"Zhang","year":"2018","journal-title":"Curr. Opin. Cell Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1593\/neo.04640","article-title":"A specific inhibitor of TGF-beta receptor kinase, SB-431542, as a potent antitumor agent for human cancers","volume":"7","author":"Halder","year":"2005","journal-title":"Neoplasia"},{"key":"ref_40","first-page":"4479","article-title":"Clinical development of galunisertib (LY2157299 monohydrate), a small molecule inhibitor of transforming growth factor-beta signaling pathway","volume":"9","author":"Herbertz","year":"2015","journal-title":"Drug Des. Devel. Ther."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1107\/S0108768101018729","article-title":"Structure and pseudosymmetry of cholesterol at 310 K","volume":"B58","author":"Hsu","year":"2002","journal-title":"Acta Cryst."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"517","DOI":"10.1038\/nature18934","article-title":"Structural basis of Smoothened regulation by its extracellular domains","volume":"535","author":"Byrne","year":"2016","journal-title":"Nature"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1176","DOI":"10.1016\/j.cell.2016.08.003","article-title":"Cellular Cholesterol Directly Activates Smoothened in Hedgehog Signaling","volume":"166","author":"Huang","year":"2016","journal-title":"Cell"},{"key":"ref_44","first-page":"46","article-title":"The prediction of human pharmacokinetic parameters from preclinical and in vitro metabolism data","volume":"283","author":"Obach","year":"1997","journal-title":"J. Pharm. Exp."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1038\/nchembio.765","article-title":"Oxysterols are allosteric activators of the oncoprotein Smoothened","volume":"8","author":"Nachtergaele","year":"2012","journal-title":"Nat. Chem. Biol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1872","DOI":"10.1002\/jbmr.2213","article-title":"A novel osteogenic oxysterol compound for therapeutic development to promote bone growth: Activation of hedgehog signaling and osteogenesis through smoothened binding","volume":"29","author":"Montgomery","year":"2014","journal-title":"J. Bone Min. Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"895","DOI":"10.1111\/tra.12177","article-title":"Binding domain-driven intracellular trafficking of sterols for synthesis of steroid hormones, bile acids and oxysterols","volume":"15","author":"Midzak","year":"2014","journal-title":"Traffic"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"10139","DOI":"10.1073\/pnas.1019441108","article-title":"Structural basis for pregnenolone biosynthesis by the mitochondrial monooxygenase system","volume":"108","author":"Strushkevich","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_49","first-page":"e1257453","article-title":"Novel TGF-\u03b2 inhibitors ready for prime time in onco-immunology","volume":"6","author":"Faivre","year":"2016","journal-title":"Oncoimmunology"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1738","DOI":"10.1126\/sciimmunol.aaj1738","article-title":"Targeting latency-associated peptide promotes antitumor immunity","volume":"2","author":"Gabriely","year":"2017","journal-title":"Sci. Immunol."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/8\/10\/1297\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:28:27Z","timestamp":1760189307000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/8\/10\/1297"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,10,22]]},"references-count":50,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2019,10]]}},"alternative-id":["cells8101297"],"URL":"https:\/\/doi.org\/10.3390\/cells8101297","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,10,22]]}}}