{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,2]],"date-time":"2025-11-02T06:50:03Z","timestamp":1762066203362,"version":"build-2065373602"},"reference-count":39,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2022,9,19]],"date-time":"2022-09-19T00:00:00Z","timestamp":1663545600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"FEDER","award":["007491","UID\/Multi\/00709","022161"],"award-info":[{"award-number":["007491","UID\/Multi\/00709","022161"]}]},{"name":"FCT\u2014Foundation for Science and Technology","award":["007491","UID\/Multi\/00709","022161"],"award-info":[{"award-number":["007491","UID\/Multi\/00709","022161"]}]},{"name":"FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC","award":["007491","UID\/Multi\/00709","022161"],"award-info":[{"award-number":["007491","UID\/Multi\/00709","022161"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>A series of novel 21E-arylidene-4-azapregn-5-ene steroids has been successfully designed, synthesized and structurally characterized, and their antiproliferative activity was evaluated in four different cell lines. Within this group, the 21E-(pyridin-3-yl)methylidene derivative exhibited significant cytotoxic activity in hormone-dependent cells LNCaP (IC50 = 10.20 \u00b5M) and T47-D cells (IC50 = 1.33 \u00b5M). In PC-3 androgen-independent cells, the steroid 21E-p-nitrophenylidene-4-azapregn-5-ene was the most potent of this series (IC50 = 3.29 \u00b5M). Considering these results, the 21E-(pyridin-3-yl)methylidene derivative was chosen for further biological studies on T47-D and LNCaP cells, and it was shown that this azasteroid seems to lead T47-D cells to apoptotic death. Finally, molecular docking studies were performed to explore the affinity of these 4-azapregnene derivatives to several steroid targets, namely 5\u03b1-reductase type 2, estrogen receptor \u03b1, androgen receptor and CYP17A1. In general, compounds presented higher affinity to 5\u03b1-reductase type 2 and estrogen receptor \u03b1.<\/jats:p>","DOI":"10.3390\/molecules27186126","type":"journal-article","created":{"date-parts":[[2022,9,20]],"date-time":"2022-09-20T04:28:55Z","timestamp":1663648135000},"page":"6126","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Novel 4-Azapregnene Derivatives as Potential Anticancer Agents: Synthesis, Antiproliferative Activity and Molecular Docking Studies"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8030-9379","authenticated-orcid":false,"given":"Vanessa","family":"Brito","sequence":"first","affiliation":[{"name":"Health Sciences Research Centre (CICS-UBI), Universidade da Beira Interior, Av. Infante D. Henrique, 6200-506 Covilh\u00e3, Portugal"}]},{"given":"Adriana Oliveira","family":"Santos","sequence":"additional","affiliation":[{"name":"Health Sciences Research Centre (CICS-UBI), Universidade da Beira Interior, Av. Infante D. Henrique, 6200-506 Covilh\u00e3, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4213-0714","authenticated-orcid":false,"given":"Gilberto","family":"Alves","sequence":"additional","affiliation":[{"name":"Health Sciences Research Centre (CICS-UBI), Universidade da Beira Interior, Av. Infante D. Henrique, 6200-506 Covilh\u00e3, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0110-4795","authenticated-orcid":false,"given":"Paulo","family":"Almeida","sequence":"additional","affiliation":[{"name":"Health Sciences Research Centre (CICS-UBI), Universidade da Beira Interior, Av. Infante D. Henrique, 6200-506 Covilh\u00e3, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4297-5108","authenticated-orcid":false,"given":"Samuel","family":"Silvestre","sequence":"additional","affiliation":[{"name":"Health Sciences Research Centre (CICS-UBI), Universidade da Beira Interior, Av. Infante D. Henrique, 6200-506 Covilh\u00e3, Portugal"},{"name":"Center for Neuroscience and Cell Biology (CNC), University of Coimbra, 3004-517 Coimbra, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2022,9,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.2174\/1389557517666171003103245","article-title":"Recent Progress of Novel Steroid Derivatives and Their Potential Biological Properties","volume":"18","author":"Ke","year":"2018","journal-title":"Mini-Reviews Med. Chem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.5402\/2012\/985495","article-title":"Applications of Steroid in Clinical Practice: A Review","volume":"2012","author":"Shaikh","year":"2012","journal-title":"ISRN Anesthesiol."},{"doi-asserted-by":"crossref","unstructured":"Brito, V., Alves, G., Almeida, P., and Silvestre, S. (2021). Highlights on Steroidal Arylidene Derivatives as a Source of Pharmacologically Active Compounds: A Review. Molecules, 26.","key":"ref_3","DOI":"10.3390\/molecules26072032"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1021\/cc060098z","article-title":"2\u03b2-(N-Substituted Piperazino)-5\u03b1-Androstane-3\u03b1,17\u03b2-Diols: Parallel Solid-Phase Synthesis and Antiproliferative Activity on Human","volume":"9","author":"Roy","year":"2007","journal-title":"J. Comb. Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"805","DOI":"10.1016\/j.steroids.2010.02.014","article-title":"Studies on Novel D-Ring Substituted Steroidal Pyrazolines as Potential Anticancer Agents","volume":"75","author":"Banday","year":"2010","journal-title":"Steroids"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1016\/j.ejmech.2013.08.016","article-title":"Synthesis and Cytotoxic Activity of Some Novel Steroidal C-17 Pyrazolinyl Derivatives","volume":"69","author":"Fan","year":"2013","journal-title":"Eur. J. Med. Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1016\/j.jsbmb.2013.04.006","article-title":"Steroidal 5\u03b1-Reductase and 17\u03b1-Hydroxylase\/17,20-Lyase (CYP17) Inhibitors Useful in the Treatment of Prostatic Diseases","volume":"137","author":"Salvador","year":"2013","journal-title":"J. Steroid Biochem. Mol. Biol."},{"key":"ref_8","first-page":"737","article-title":"Synthesis of 17b-N-Arylcarbamoylandrost-4-En-3-One Derivatives and Their Antiproliferative Effect on Human Androgen-Sensitive LNCaP Cell Line","volume":"4","author":"Cabeza","year":"2016","journal-title":"Eur. J. Med. Chem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"7257","DOI":"10.1016\/j.bmc.2013.09.063","article-title":"17(E)-Picolinylidene Androstane Derivatives as Potential Inhibitors of Prostate Cancer Cell Growth: Antiproliferative Activity and Molecular Docking Studies","volume":"21","author":"Ajdukovic","year":"2013","journal-title":"Bioorg. Med. Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1391","DOI":"10.1016\/j.steroids.2008.07.004","article-title":"Synthesis of 16E-[3-Methoxy-4-(2-Aminoethoxy)Benzylidene]Androstene Derivatives as Potent Cytotoxic Agents","volume":"73","author":"Bansal","year":"2008","journal-title":"Steroids"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.bioorg.2012.08.005","article-title":"Synthesis and Aromatase Inhibitory Activity of Some New 16E-Arylidenosteroids","volume":"45","author":"Bansal","year":"2012","journal-title":"Bioorg. Chem."},{"key":"ref_12","first-page":"551","article-title":"Synthesis and Cytotoxic Studies of Hydroximino Derivatives of Some 16E-Arylidenosteroids","volume":"556","author":"Chattopadhaya","year":"2004","journal-title":"Arzneim. Forshung - Drug Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1529","DOI":"10.1002\/cbdv.200490113","article-title":"Synthesis and in Vitro Antineoplastic Evaluation of Certain 16-(4-Substituted Benzylidene) Derivatives of Androst-5-Ene","volume":"1","author":"Dubey","year":"2004","journal-title":"Chem. Biodivers."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1554","DOI":"10.1016\/j.steroids.2011.09.006","article-title":"Pregnenolone Derivatives as Potential Anticancer Agents","volume":"76","author":"Choudhary","year":"2011","journal-title":"Steroids"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.steroids.2014.09.004","article-title":"Steroidal Pyrazolines and Pyrazoles as Potential 5\u03b1-Reductase Inhibitors: Synthesis and Biological Evaluation","volume":"92","author":"Banday","year":"2014","journal-title":"Steroids"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/j.steroids.2014.03.010","article-title":"Benzylidine Pregnenolones and Their Oximes as Potential Anticancer Agents: Synthesis and Biological Evaluation","volume":"84","author":"Banday","year":"2014","journal-title":"Steroids"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.steroids.2017.04.006","article-title":"Synthesis of Novel 4\u2032-Acylamino Modified 21E-Benzylidene Steroidal Derivatives and Their Cytotoxic Activities","volume":"123","author":"Fan","year":"2017","journal-title":"Steroids"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"6637","DOI":"10.1016\/j.bmcl.2009.10.019","article-title":"Synthesis of 3b, 7a, 11a-Trihydroxy-Pregn-21-Benzylidene-5-En-20-One Derivatives and Their Cytotoxic Activities","volume":"19","author":"Shan","year":"2009","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.steroids.2013.12.004","article-title":"Synthesis and Anticancer Activity of Novel C6-Piperazine Substituted Purine Steroid-Nucleosides Analogues","volume":"82","author":"Huang","year":"2014","journal-title":"Steroids"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/j.crci.2018.07.011","article-title":"Novel 4-Azaandrostenes as Prostate Cancer Cell Growth Inhibitors: Synthesis, Antiproliferative Effects and Molecular Docking Studies","volume":"22","author":"Brito","year":"2018","journal-title":"Comptes Rendus Chim."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"5430","DOI":"10.1038\/s41467-020-19249-z","article-title":"Structure of Human Steroid 5\u03b1-Reductase 2 with the Anti-Androgen Drug Finasteride","volume":"11","author":"Xiao","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4568","DOI":"10.1007\/s00044-012-0456-5","article-title":"Design of Potent Human Steroid 5\u03b1-Reductase Inhibitors: 3D-QSAR CoMFA, CoMSIA and Docking Studies","volume":"22","author":"Kumar","year":"2013","journal-title":"Med. Chem. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1016\/j.bmcl.2010.10.112","article-title":"Discovery of a Novel Hybrid from Finasteride and Epristeride as 5a-Reductase Inhibitor","volume":"21","author":"Yao","year":"2011","journal-title":"Bioorganic Med. Chem. Lett."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1016\/j.steroids.2008.01.022","article-title":"A Microwave Promoted and Lewis Acid Catalysed Solventless Approach to 4-Azasteroids","volume":"73","author":"Borthakur","year":"2008","journal-title":"Steroids"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1209","DOI":"10.1016\/0968-0896(96)00109-5","article-title":"Synthesis of 4,17-Diazasteroid Inhibitors of Human 5\u03b1-Reductase","volume":"4","author":"Morzycki","year":"1996","journal-title":"Bioorg. Med. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"552","DOI":"10.1016\/j.steroids.2012.01.020","article-title":"Synthesis and Antileukemic Activity of 16E-[4-(2-Carboxy)Ethoxybenzylidene]-Androstene Amides","volume":"77","author":"Bansal","year":"2012","journal-title":"Steroids"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"874","DOI":"10.1016\/j.steroids.2013.04.016","article-title":"Synthesis and Cytotoxicity of Some Novel 21E-Benzylidene Steroidal Derivatives","volume":"78","author":"Fan","year":"2013","journal-title":"Steroids"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1177\/1747519819851706","article-title":"Synthesis and Antiproliferative Activity of Novel 4-Azasteroidal-17-Hydrazone Derivatives","volume":"43","author":"Chen","year":"2019","journal-title":"J. Chem. Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"566","DOI":"10.1016\/j.steroids.2012.02.001","article-title":"Synthesis of D-Ring-Substituted (5\u2019R)- and (5\u2019S)-17b-Pyrazolinylandrostene Epimers and Comparison of Their Potential Anticancer Activities","volume":"77","author":"Huber","year":"2012","journal-title":"Steroids"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1016\/j.ejmech.2017.11.070","article-title":"Trisubstituted Barbiturates and Thiobarbiturates: Synthesis and Biological Evaluation as Xanthine Oxidase Inhibitors, Antioxidants, Antibacterial and Anti-Proliferative Agents","volume":"143","author":"Figueiredo","year":"2018","journal-title":"Eur. J. Med. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.jsbmb.2017.06.001","article-title":"Multicomponent Access to Androstano-Arylpyrimidines under Microwave Conditions and Evaluation of Their Anti-Cancer Activity in Vitro","volume":"172","author":"Baji","year":"2017","journal-title":"J. Steroid Biochem. Mol. Biol."},{"doi-asserted-by":"crossref","unstructured":"de la Guardia, C., Stephens, D.E., Dang, H.T., Quijada, M., Larionov, O.V., and Lleonart, R. (2018). Antiviral Activity of Novel Quinoline Derivatives against Dengue Virus Serotype 2. Molecules, 23.","key":"ref_32","DOI":"10.3390\/molecules23030672"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1746-1596-9-92","article-title":"Objective Assessment of Changes in Nuclear Morphology and Cell Distribution Following Induction of Apoptosis","volume":"9","author":"Eidet","year":"2014","journal-title":"Diagn. Pathol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"23996","DOI":"10.18632\/oncotarget.15098","article-title":"Caspase-9: Structure, Mechanisms and Clinical Application","volume":"8","author":"Li","year":"2017","journal-title":"Oncotarget"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1165","DOI":"10.1002\/cyto.a.20634","article-title":"Simultaneous Evaluation of Viability and Bcl-2 in Small-Cell Lung Cancer","volume":"73","author":"Santos","year":"2008","journal-title":"Cytom. Part A"},{"doi-asserted-by":"crossref","unstructured":"Armarego, W.L.F. (2017). Chapter 3 - Purification of Organic Chemicals. Purification of Laboratory Chemicals (Eighth Edition), Butterworth-Heinemann.","key":"ref_36","DOI":"10.1016\/B978-0-12-805457-4.50003-3"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1016\/S1385-299X(02)00146-0","article-title":"A Rapid Method for Obtaining Finasteride, a 5\u03b1-Reductase Inhibitor, from Commercial Tablets","volume":"9","author":"Trapani","year":"2002","journal-title":"Brain Res. Protoc."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"7593","DOI":"10.1016\/j.tet.2015.07.077","article-title":"Cell Proliferation Effects of Calix[4]Arene Derivatives","volume":"71","author":"Santos","year":"2015","journal-title":"Tetrahedron"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1639","DOI":"10.1002\/(SICI)1096-987X(19981115)19:14<1639::AID-JCC10>3.0.CO;2-B","article-title":"Automated Docking Using a Lamarckian Genetic Algorithm and an Empirical Binding Free Energy Function","volume":"19","author":"Morris","year":"1998","journal-title":"J. Comput. Chem."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/27\/18\/6126\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:34:29Z","timestamp":1760142869000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/27\/18\/6126"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,9,19]]},"references-count":39,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2022,9]]}},"alternative-id":["molecules27186126"],"URL":"https:\/\/doi.org\/10.3390\/molecules27186126","relation":{},"ISSN":["1420-3049"],"issn-type":[{"type":"electronic","value":"1420-3049"}],"subject":[],"published":{"date-parts":[[2022,9,19]]}}}