Thiosemicarbazones (TSCs) are an interesting class of ligands that show a diverse range of biological activity, including anti-fungal, anti-viral and anti-cancer effects. Our previous studies have demonstrated the potent in vivo anti-tumor activity of novel TSCs and their ability to overcome resistance to clinically used chemotherapeutics. In the current study, 35 novel TSCs of 6 different classes were designed using a combination of retro-fragments that appear in other TSCs. Additionally, di-substitution at the terminal N4 atom, which was previously identified to be critical for potent anti-cancer activity, was preserved through the incorporation of an N4-based piperazine or morpholine ring. The anti-proliferative activity of the novel TSCs were examined in a variety of cancer and normal cell-types. In particular, compounds 1d and 3c demonstrated the greatest promise as anti-cancer agents with potent and selective anti-proliferative activity. Structure-activity relationship studies revealed that the chelators that utilized "soft" donor atoms, such as nitrogen and sulfur, resulted in potent anti-cancer activity. Indeed, the N,N,S donor atom set was crucial for the formation of redox active iron complexes that were able to mediate the oxidation of ascorbate. This further highlights the important role of reactive oxygen species generation in mediating potent anti-cancer activity. Significantly, this study identified the potent and selective anti-cancer activity of 1d and 3c that warrants further examination.

A Chełkowski Institute of Physics and Silesian Interdisciplinary Centre for Education and Research University of Silesia Katowice Silesia Poland

Department of Biochemical Sciences Charles University Prague Faculty of Pharmacy in Hradec Králové Hradec Králové Czech Republic

Department of Immunology Medical University of Warsaw Warsaw Mazovia Poland

Department of Immunology Medical University of Warsaw Warsaw Mazovia Poland; Institute of Physical Chemistry Polish Academy of Sciences Warsaw Mazovia Poland

Department of Pathology and Bosch Institute University of Sydney Sydney New South Wales Australia

Institute of Chemistry University of Silesia Katowice Silesia Poland

Institute of Chemistry University of Silesia Katowice Silesia Poland; A Chełkowski Institute of Physics and Silesian Interdisciplinary Centre for Education and Research University of Silesia Katowice Silesia Poland

Zobrazit více v PubMed

Kalinowski DS, Richardson DR (2005) The evolution of iron chelators for the treatment of iron overload disease and cancer. Pharmacol Rev 57: 547–583. PubMed

Lieu PT, Heiskala M, Peterson PA, Yang Y (2001) The roles of iron in health and disease. Mol Aspects Med 22: 1–87. PubMed

Merlot AM, Kalinowski DS, Richardson DR (2013) Novel chelators for cancer treatment: where are we now? Antioxid Redox Signal 18: 973–1006. PubMed

Kolberg M, Strand KR, Graff P, Andersson KK (2004) Structure, function, and mechanism of ribonucleotide reductases. Biochim Biophys Acta 1699: 1–34. PubMed

Thelander L, Reichard P (1979) Reduction of ribonucleotides. Annu Rev Biochem 48: 133–158. PubMed

Richardson D, Baker E (1992) Two mechanisms of iron uptake from transferrin by melanoma cells. The effect of desferrioxamine and ferric ammonium citrate. J Biol Chem 267: 13972–13979. PubMed

Richardson DR, Baker E (1990) The uptake of iron and transferrin by the human malignant melanoma cell. Biochim Biophys Acta 1053: 1–12. PubMed

Trinder D, Zak O, Aisen P (1996) Transferrin receptor-independent uptake of differic transferrin by human hepatoma cells with antisense inhibition of receptor expression. Hepatology 23: 1512–1520. PubMed

Elford HL, Freese M, Passamani E, Morris HP (1970) Ribonucleotide reductase and cell proliferation. I. Variations of ribonucleotide reductase activity with tumor growth rate in a series of rat hepatomas. J Biol Chem 245: 5228–5233. PubMed

Kalinowski DS, Yu Y, Sharpe PC, Islam M, Liao YT, et al. (2007) Design, synthesis, and characterization of novel iron chelators: structure-activity relationships of the 2-benzoylpyridine thiosemicarbazone series and their 3-nitrobenzoyl analogues as potent antitumor agents. J Med Chem 50: 3716–3729. PubMed

Kunos C, Radivoyevitch T, Abdul-Karim FW, Fanning J, Abulafia O, et al. (2012) Ribonucleotide reductase inhibition restores platinum-sensitivity in platinum-resistant ovarian cancer: a Gynecologic Oncology Group Study. J Transl Med 10: 79. PubMed PMC

Lovejoy DB, Sharp DM, Seebacher N, Obeidy P, Prichard T, et al. (2012) Novel second-generation di-2-pyridylketone thiosemicarbazones show synergism with standard chemotherapeutics and demonstrate potent activity against lung cancer xenografts after oral and intravenous administration in vivo. J Med Chem 55: 7230–7244. PubMed

Lukmantara AY, Kalinowski DS, Kumar N, Richardson DR (2013) Synthesis and biological evaluation of substituted 2-benzoylpyridine thiosemicarbazones: novel structure-activity relationships underpinning their anti-proliferative and chelation efficacy. Bioorg Med Chem Lett 23: 967–974. PubMed

Richardson DR, Kalinowski DS, Richardson V, Sharpe PC, Lovejoy DB, et al. (2009) 2-Acetylpyridine thiosemicarbazones are potent iron chelators and antiproliferative agents: redox activity, iron complexation and characterization of their antitumor activity. J Med Chem 52: 1459–1470. PubMed

Richardson DR, Sharpe PC, Lovejoy DB, Senaratne D, Kalinowski DS, et al. (2006) Dipyridyl thiosemicarbazone chelators with potent and selective antitumor activity form iron complexes with redox activity. J Med Chem 49: 6510–6521. PubMed

Serda M, Kalinowski DS, Mrozek-Wilczkiewicz A, Musiol R, Szurko A, et al. (2012) Synthesis and characterization of quinoline-based thiosemicarbazones and correlation of cellular iron-binding efficacy to anti-tumor efficacy. Bioorg Med Chem Lett 22: 5527–5531. PubMed

Stefani C, Jansson PJ, Gutierrez E, Bernhardt PV, Richardson DR, et al. (2013) Alkyl substituted 2′-benzoylpyridine thiosemicarbazone chelators with potent and selective anti-neoplastic activity: novel ligands that limit methemoglobin formation. J Med Chem 56: 357–370. PubMed

Stefani C, Punnia-Moorthy G, Lovejoy DB, Jansson PJ, Kalinowski DS, et al. (2011) Halogenated 2′-benzoylpyridine thiosemicarbazone (XBpT) chelators with potent and selective anti-neoplastic activity: relationship to intracellular redox activity. J Med Chem 54: 6936–6948. PubMed

Yu Y, Kalinowski DS, Kovacevic Z, Siafakas AR, Jansson PJ, et al. (2009) Thiosemicarbazones from the old to new: iron chelators that are more than just ribonucleotide reductase inhibitors. J Med Chem 52: 5271–5294. PubMed

Yu Y, Suryo Rahmanto Y, Richardson DR (2012) Bp44mT: an orally active iron chelator of the thiosemicarbazone class with potent anti-tumour efficacy. Br J Pharmacol 165: 148–166. PubMed PMC

Yuan J, Lovejoy DB, Richardson DR (2004) Novel di-2-pyridyl-derived iron chelators with marked and selective antitumor activity: in vitro and in vivo assessment. Blood 104: 1450–1458. PubMed

Chaston TB, Lovejoy DB, Watts RN, Richardson DR (2003) Examination of the antiproliferative activity of iron chelators: multiple cellular targets and the different mechanism of action of triapine compared with desferrioxamine and the potent pyridoxal isonicotinoyl hydrazone analogue 311. Clin Cancer Res 9: 402–414. PubMed

Lovejoy DB, Jansson PJ, Brunk UT, Wong J, Ponka P, et al. (2011) Antitumor activity of metal-chelating compound Dp44mT is mediated by formation of a redox-active copper complex that accumulates in lysosomes. Cancer Res 71: 5871–5880. PubMed

Shao J, Zhou B, Di Bilio AJ, Zhu L, Wang T, et al. (2006) A Ferrous-Triapine complex mediates formation of reactive oxygen species that inactivate human ribonucleotide reductase. Mol Cancer Ther 5: 586–592. PubMed

Zhu L, Zhou B, Chen X, Jiang H, Shao J, et al. (2009) Inhibitory mechanisms of heterocyclic carboxaldehyde thiosemicabazones for two forms of human ribonucleotide reductase. Biochem Pharmacol 78: 1178–1185. PubMed

Kovacevic Z, Chikhani S, Lovejoy DB, Richardson DR (2011) Novel thiosemicarbazone iron chelators induce up-regulation and phosphorylation of the metastasis suppressor N-myc down-stream regulated gene 1: a new strategy for the treatment of pancreatic cancer. Mol Pharmacol 80: 598–609. PubMed

Kovacevic Z, Chikhani S, Lui GY, Sivagurunathan S, Richardson DR (2013) The iron-regulated metastasis suppressor NDRG1 targets NEDD4L, PTEN, and SMAD4 and inhibits the PI3K and Ras signaling pathways. Antioxid Redox Signal 18: 874–887. PubMed

Kovacevic Z, Sivagurunathan S, Mangs H, Chikhani S, Zhang D, et al. (2011) The metastasis suppressor, N-myc downstream regulated gene 1 (NDRG1), upregulates p21 via p53-independent mechanisms. Carcinogenesis 32: 732–740. PubMed

Le NT, Richardson DR (2004) Iron chelators with high antiproliferative activity up-regulate the expression of a growth inhibitory and metastasis suppressor gene: a link between iron metabolism and proliferation. Blood 104: 2967–2975. PubMed

Bernhardt PV, Sharpe PC, Islam M, Lovejoy DB, Kalinowski DS, et al. (2009) Iron chelators of the dipyridylketone thiosemicarbazone class: precomplexation and transmetalation effects on anticancer activity. J Med Chem 52: 407–415. PubMed

Feun L, Modiano M, Lee K, Mao J, Marini A, et al. (2002) Phase I and pharmacokinetic study of 3-aminopyridine-2-carboxaldehyde thiosemicarbazone (3-AP) using a single intravenous dose schedule. Cancer Chemother Pharmacol 50: 223–229. PubMed

Karp JE, Giles FJ, Gojo I, Morris L, Greer J, et al. (2008) A phase I study of the novel ribonucleotide reductase inhibitor 3-aminopyridine-2-carboxaldehyde thiosemicarbazone (3-AP, Triapine) in combination with the nucleoside analog fludarabine for patients with refractory acute leukemias and aggressive myeloproliferative disorders. Leuk Res 32: 71–77. PubMed PMC

Kunos CA, Radivoyevitch T, Waggoner S, Debernardo R, Zanotti K, et al. (2013) Radiochemotherapy plus 3-aminopyridine-2-carboxaldehyde thiosemicarbazone (3-AP, NSC #663249) in advanced-stage cervical and vaginal cancers. Gynecol Oncol 130: 75–80. PubMed PMC

Kunos CA, Waggoner S, von Gruenigen V, Eldermire E, Pink J, et al. (2010) Phase I trial of pelvic radiation, weekly cisplatin, and 3-aminopyridine-2-carboxaldehyde thiosemicarbazone (3-AP, NSC #663249) for locally advanced cervical cancer. Clin Cancer Res 16: 1298–1306. PubMed PMC

Ma B, Goh BC, Tan EH, Lam KC, Soo R, et al. (2008) A multicenter phase II trial of 3-aminopyridine-2-carboxaldehyde thiosemicarbazone (3-AP, Triapine) and gemcitabine in advanced non-small-cell lung cancer with pharmacokinetic evaluation using peripheral blood mononuclear cells. Invest New Drugs 26: 169–173. PubMed

Mackenzie MJ, Saltman D, Hirte H, Low J, Johnson C, et al. (2007) A Phase II study of 3-aminopyridine-2-carboxaldehyde thiosemicarbazone (3-AP) and gemcitabine in advanced pancreatic carcinoma. A trial of the Princess Margaret hospital Phase II consortium. Invest New Drugs 25: 553–558. PubMed

Nutting CM, van Herpen CM, Miah AB, Bhide SA, Machiels JP, et al. (2009) Phase II study of 3-AP Triapine in patients with recurrent or metastatic head and neck squamous cell carcinoma. Ann Oncol 20: 1275–1279. PubMed

Ocean AJ, Christos P, Sparano JA, Matulich D, Kaubish A, et al. (2010) Phase II trial of the ribonucleotide reductase inhibitor 3-aminopyridine-2-carboxaldehydethiosemicarbazone plus gemcitabine in patients with advanced biliary tract cancer. Cancer Chemother Pharmacol 68: 379–388. PubMed PMC

Odenike OM, Larson RA, Gajria D, Dolan ME, Delaney SM, et al. (2008) Phase I study of the ribonucleotide reductase inhibitor 3-aminopyridine-2-carboxaldehyde-thiosemicarbazone (3-AP) in combination with high dose cytarabine in patients with advanced myeloid leukemia. Invest New Drugs 26: 233–239. PubMed PMC

Wadler S, Makower D, Clairmont C, Lambert P, Fehn K, et al. (2004) Phase I and pharmacokinetic study of the ribonucleotide reductase inhibitor, 3-aminopyridine-2-carboxaldehyde thiosemicarbazone, administered by 96-hour intravenous continuous infusion. J Clin Oncol 22: 1553–1563. PubMed

Yen Y, Margolin K, Doroshow J, Fishman M, Johnson B, et al. (2004) A phase I trial of 3-aminopyridine-2-carboxaldehyde thiosemicarbazone in combination with gemcitabine for patients with advanced cancer. Cancer Chemother Pharmacol 54: 331–342. PubMed

Whitnall M, Howard J, Ponka P, Richardson DR (2006) A class of iron chelators with a wide spectrum of potent antitumor activity that overcomes resistance to chemotherapeutics. Proc Natl Acad Sci U S A 103: 14901–14906. PubMed PMC

Mrozek-Wilczkiewicz A, Serda M, Musiol R, Malecki G, Szurko A, et al. (2014) Iron chelators in photodynamic therapy revisited: Synergistic effect by novel highly active thiosemicarbazones. ACS Med Chem Lett 5: 336–339. PubMed PMC

Serda M, Mrozek-Wilczkiewicz A, Jampilek J, Pesko M, Kralova K, et al. (2012) Investigation of the biological properties of (hetero)aromatic thiosemicarbazones. Molecules 17: 13483–13502. PubMed PMC

Hu WX, Zhou W, Xia CN, Wen X (2006) Synthesis and anticancer activity of thiosemicarbazones. Bioorg Med Chem Lett 16: 2213–2218. PubMed

Stanojkovic TP, Kovala-Demertzi D, Primikyri A, Garcia-Santos I, Castineiras A, et al. (2010) Zinc(II) complexes of 2-acetyl pyridine 1-(4-fluorophenyl)-piperazinyl thiosemicarbazone: Synthesis, spectroscopic study and crystal structures - potential anticancer drugs. J Inorg Biochem 104: 467–476. PubMed

Ghose AK, Crippen GM (1987) Atomic physicochemical parameters for three-dimensional-structure-directed quantitative structure−activity relationships. 2. Modeling dispersive and hydrophobic interactions. J Chem Inf Comput Sci 27: 21–35. PubMed

Viswanadhan VN, Ghose AK, Revankar GR, Robins RK (1987) Atomic physiochemical parameters for 3-dimensional-structure directed quantitative structure−activity relationships 4. Additional parameters for hydrophobic and dispersive interactions and their application for an automated superposition of certain naturally-occuring nucleoside antibiotics. J Chem Inf Comput Sci 29: 163–172.

Broto P, Moreau G, Vandycke C (1984) Molecular structures: Perception, autocorrelation descriptor and SAR studies. System of atomic contributions for the calculation of the n-octanol/water partition coefficients. Eur J Med Chem Chim Theor 19: 71–78.

Richardson DR, Tran EH, Ponka P (1995) The potential of iron chelators of the pyridoxal isonicotinoyl hydrazone class as effective antiproliferative agents. Blood 86: 4295–4306. PubMed

Baker E, Richardson D, Gross S, Ponka P (1992) Evaluation of the iron chelation potential of hydrazones of pyridoxal, salicylaldehyde and 2-hydroxy-1-naphthylaldehyde using the hepatocyte in culture. Hepatology 15: 492–501. PubMed

Kalinowski DS, Sharpe PC, Bernhardt PV, Richardson DR (2007) Design, synthesis, and characterization of new iron chelators with anti-proliferative activity: structure-activity relationships of novel thiohydrazone analogues. J Med Chem 50: 6212–6225. PubMed

Kalinowski DS, Sharpe PC, Bernhardt PV, Richardson DR (2008) Structure-activity relationships of novel iron chelators for the treatment of iron overload disease: the methyl pyrazinylketone isonicotinoyl hydrazone series. J Med Chem 51: 331–344. PubMed

Hann MM, Keseru GM (2012) Finding the sweet spot: the role of nature and nurture in medicinal chemistry. Nat Rev Drug Discov 11: 355–365. PubMed

Meanwell NA (2011) Improving drug candidates by design: a focus on physicochemical properties as a means of improving compound disposition and safety. Chem Res Toxicol 24: 1420–1456. PubMed

Gleeson MP (2008) Generation of a set of simple, interpretable ADMET rules of thumb. J Med Chem 51: 817–834. PubMed

Hann MM (2011) Molecular obesity, potency and other addictions in drug discovery. Med Chem Commun 2: 349–355.

Proschak E, Tanrikulu Y, Schneider G (2008) Chapter 7: Fragment-based de novo design of drug-like molecules. In: Varnek A, Tropsha A, editors.Chemoinformatics approaches to virtual screening.Cambridge: Royal Society of Chemistry. pp.217–239.

Becker EM, Lovejoy DB, Greer JM, Watts R, Richardson DR (2003) Identification of the di-pyridyl ketone isonicotinoyl hydrazone (PKIH) analogues as potent iron chelators and anti-tumour agents. Br J Pharmacol 138: 819–830. PubMed PMC

Wang Y, Ai J, Chen Y, Wang L, Liu G, et al. (2011) Synthesis and c-Met kinase inhibition of 3,5-disubstituted and 3,5,7-trisubstituted quinolines: identification of 3-(4-acetylpiperazin-1-yl)-5-(3-nitrobenzylamino)-7- (trifluoromethyl)quinoline as a novel anticancer agent. J Med Chem 54: 2127–2142. PubMed

Chetan B, Bunha M, Jagrat M, Sinha BN, Saiko P, et al. (2010) Design, synthesis and anticancer activity of piperazine hydroxamates and their histone deacetylase (HDAC) inhibitory activity. Bioorg Med Chem Lett 20: 3906–3910. PubMed

Hou X, Ge Z, Wang T, Guo W, Cui J, et al. (2006) Dithiocarbamic acid esters as anticancer agent. Part 1: 4-substituted-piperazine-1-carbodithioic acid 3-cyano-3,3-diphenyl-propyl esters. Bioorg Med Chem Lett 16: 4214–4219. PubMed

Yu K, Toral-Barza L, Shi C, Zhang WG, Lucas J, et al. (2009) Biochemical, cellular, and in vivo activity of novel ATP-competitive and selective inhibitors of the mammalian target of rapamycin. Cancer Res 69: 6232–6240. PubMed

Gao H, Yamasaki EF, Chan KK, Shen LL, Snapka RM (2003) DNA sequence specificity for topoisomerase II poisoning by the quinoxaline anticancer drugs XK469 and CQS. Mol Pharmacol 63: 1382–1388. PubMed

Silva JL, Gallo CV, Costa DC, Rangel LP (2014) Prion-like aggregation of mutant p53 in cancer. Trends Biochem Sci In Press April 25, 2014. PubMed

Breen L, Heenan M, Amberger-Murphy V, Clynes M (2007) Investigation of the role of p53 in chemotherapy resistance of lung cancer cell lines. Anticancer Res 27: 1361–1364. PubMed

Whibley C, Pharoah PD, Hollstein M (2009) p53 polymorphisms: cancer implications. Nat Rev Cancer 9: 95–107. PubMed

Mladenka P, Kalinowski DS, Haskova P, Bobrovova Z, Hrdina R, et al. (2009) The novel iron chelator, 2-pyridylcarboxaldehyde 2-thiophenecarboxyl hydrazone, reduces catecholamine-mediated myocardial toxicity. Chem Res Toxicol 22: 208–217. PubMed

Deraeve C, Pitie M, Meunier B (2006) Influence of chelators and iron ions on the production and degradation of H2O2 by beta-amyloid-copper complexes. J Inorg Biochem 100: 2117–2126. PubMed

Di Vaira M, Bazzicalupi C, Orioli P, Messori L, Bruni B, et al. (2004) Clioquinol, a drug for Alzheimer's disease specifically interfering with brain metal metabolism: Structural characterization of Its zinc(II) and copper(II) complexes. Inorg Chem 43: 3795–3797. PubMed

Lovejoy DB, Richardson DR (2002) Novel "hybrid" iron chelators derived from aroylhydrazones and thiosemicarbazones demonstrate selective antiproliferative activity against tumor cells. Blood 100: 666–676. PubMed

Bernhardt PV, Wilson GJ, Sharpe PC, Kalinowski DS, Richardson DR (2008) Tuning the antiproliferative activity of biologically active iron chelators: characterization of the coordination chemistry and biological efficacy of 2-acetylpyridine and 2-benzoylpyridine hydrazone ligands. J Biol Inorg Chem 13: 107–119. PubMed

Chemistry towards Biology-Instruct: Snapshot

Iron-Chelation Treatment by Novel Thiosemicarbazone Targets Major Signaling Pathways in Neuroblastoma

Design and synthesis of anticancer 1-hydroxynaphthalene-2-carboxanilides with a p53 independent mechanism of action

The Forty-Sixth Euro Congress on Drug Synthesis and Analysis: Snapshot †