title: Cyclohexa-2,5-diene-1,4-dione-based antiproliferative agents: design, synthesis, and cytotoxic evaluation.
abstract: BACKGROUND
  Tumors are diseases characterized by uncontrolled cell growth and, in spite of the progress of medicine over the years, continue to represent a major threat to the health, requiring new therapies. Several synthetic compounds, such as those derived from natural sources, have been identified as anticancer drugs; among these compounds quinone represent the second largest class of anticancer agents in use. Several studies have shown that these act on tumor cells through several mechanisms. An important objective of this work is to develop quinoidscompounds showing antitumor activity, but with fewer side effects. The parachinone cannabinol HU-331, is a small molecule that with its core 4-hydroxy-1,4-benzoquinone, exhibits a potent and selective cytotoxic activity on different tumor cell lines. A series of derivatives 3-hydroxy-1,4-benzochinoni were thus developed through HU-331 chemical modifications. The purpose of the work is to test the ability of the compounds to induce proliferative inhibition and study the mechanisms of cell death.
  
  
  METHODS
  The antitumor activities were evaluated in vitro by examining their cytotoxic effects against different human cancer cell lines. All cell lines tested were plated in 96-multiwell and treated with HU-100-V at different concentrations and cell viability was evaluated byMTT assay. Subsequently via flow cytometry (FACS) it was possible to assess apoptosis by the system of double labeling with PI and Annexin-V, and the effect of the compounds on ROS formation by measuring the dichlorofluorescein fluorescence.
  
  
  RESULTS
  The substitution by n-hexyl chain considerably enhanced the bioactivity of the compounds. In details, 2-hexyl-5-hydroxycyclohexa-2,5-diene-1,4-dione (V), 2,5-Dimethoxy-3-hexyl-2,5-cyclohexadiene-1,4-dione (XII) and 2-hydroxy-5-methoxy-3-hexyl-cyclohexa-2,5-diene-1,4-dione (XIII) showed most prominent cytotoxicity against almost human tumour cell lines. Compound V was further subjected to downstream apoptotic analysis, demostrating a time-dependent pro-apoptotic activity on human melanoma M14 cell line mediated by caspases activation and poly-(ADP-ribose)-polymerase (PARP) protein cleavage.
  
  
  CONCLUSIONS
  These findings indicate that 2-hexyl-5-idrossicicloesa-2,5-diene-1,4-dione can be a promising compound for the design of a new class of antineoplastic derivatives.Carmen Petronzi, Michela Festa, Antonella Peduto and Maria Castellano: equally contributed equally to this work.
authors: Department of Pharmaceutical and Biomedical Sciences, University of Salerno, via Ponte Don Melillo, Fisciano, SA, 84084, Italy.
authors: Carmen
authors: C
authors: Petronzi
authors: Michela
authors: M
authors: Festa
authors: Antonella
authors: A
authors: Peduto
authors: Maria
authors: M
authors: Castellano
authors: Jessica
authors: J
authors: Marinello
authors: Antonio
authors: A
authors: Massa
authors: Anna
authors: A
authors: Capasso
authors: Giovanni
authors: G
authors: Capranico
authors: Annalisa
authors: A
authors: La Gatta
authors: Mario
authors: M
authors: De Rosa
authors: Michele
authors: M
authors: Caraglia
authors: Rosanna
authors: R
authors: Filosa
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry* / Q000494:pharmacology* / Q000633:toxicity; D017209:Apoptosis / Q000187:drug effects; D016227:Benzoquinones / Q000737:chemistry* / Q000494:pharmacology* / Q000633:toxicity; D002185:Cannabidiol / Q000031:analogs & derivatives / Q000494:pharmacology; D020169:Caspases / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D006801:Humans; D020128:Inhibitory Concentration 50; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D059003:Topoisomerase Inhibitors / Q000737:chemistry / Q000494:pharmacology / Q000633:toxicity
chemical_list: D000970:Antineoplastic Agents; D016227:Benzoquinones; D017382:Reactive Oxygen Species; D059003:Topoisomerase Inhibitors; C073718:cannabidiol hydroxyquinone; D002185:Cannabidiol; C004532:quinone; D020169:Caspases
references: J Exp Clin Cancer Res. 2010 Sep 10;29:124
references: 20831794
references: Genes Dev. 1999 Feb 1;13(3):239-52
references: 9990849
references: J Biol Chem. 1999 Mar 26;274(13):8516-23
references: 10085084
references: Anticancer Drugs. 1994 Dec;5(6):645-9
references: 7888702
references: J Exp Clin Cancer Res. 2012 Feb 08;31:12
references: 22316341
references: J Exp Clin Cancer Res. 2012 Jul 19;31:60
references: 22812382
references: Nat Protoc. 2006;1(3):1458-61
references: 17406435
references: J Leukoc Biol. 1990 May;47(5):440-8
references: 2159514
references: Front Biosci. 2000 Jul 01;5:D629-38
references: 10877994
references: J Med Chem. 2004 Jul 15;47(15):3800-6
references: 15239658
references: Prostate. 2013 Jan;73(2):133-45
references: 22674285
references: Mol Cancer Ther. 2007 Jan;6(1):173-83
references: 17237277
references: Cancer Res. 2000 Jul 15;60(14):3770-6
references: 10919649
references: Ann Surg. 2007 Aug;246(2):259-68
references: 17667505
references: J Nat Prod. 2001 Jan;64(1):2-5
references: 11170656
references: J Med Chem. 2004 May 6;47(10):2430-40
references: 15115387
references: Bioorg Med Chem. 2009 Jan 1;17(1):13-24
references: 19058969
references: Mini Rev Med Chem. 2010 May;10(5):436-54
references: 20370705
references: Eur J Med Chem. 2011 Feb;46(2):488-96
references: 21163556
references: J Immunol Methods. 1983 Dec 16;65(1-2):55-63
references: 6606682
references: J Pharmacol Exp Ther. 2007 Aug;322(2):646-53
references: 17478614
references: Int J Cancer. 1995 May 4;61(3):342-7
references: 7729946
references: Phytother Res. 2003 Sep;17(8):887-91
references: 13680818
references: Chemotherapy. 1994 Mar-Apr;40(2):109-13
references: 7510605
references: Eur J Med Chem. 2007 Mar;42(3):293-306
references: 17254669
references: Mol Pharmacol. 2006 Jul;70(1):51-9
references: 16571653
references: Biotechnol Adv. 2012 Jan-Feb;30(1):169-84
references: 21871555
references: Bioorg Med Chem. 2011 Nov 1;19(21):6419-29
references: 21944546
references: Med Oncol. 2011 Dec;28(4):1584-8
references: 20625944
references: Cell. 2001 Mar 9;104(5):791-800
references: 11257232
references: Phytochemistry. 2002 Dec;61(8):923-6
references: 12453518