Cytotoxic evaluation of 3-aminopyridine-2-carboxaldehyde thiosemicarbazone in peripheral blood lymphocytes of patients with refractory solid tumors using electron paramagnetic resonance

Kolesar,Jill  M.; Sachidanandam,Kamakshi; Schelman,William  R.; Eickhoff,Jens; Holen,Kyle  D.; Traynor,Anne  M.; Alberti,Dona  B.; Thomas,James  P.; Chitambar,Christopher  R.; Wilding,George; Antholine,William  E.

doi:10.3892/etm.2010.165

Cytotoxic evaluation of 3-aminopyridine-2-carboxaldehyde thiosemicarbazone in peripheral blood lymphocytes of patients with refractory solid tumors using electron paramagnetic resonance

Authors:
- Jill M. Kolesar
- Kamakshi Sachidanandam
- William R. Schelman
- Jens Eickhoff
- Kyle D. Holen
- Anne M. Traynor
- Dona B. Alberti
- James P. Thomas
- Christopher R. Chitambar
- George Wilding
- William E. Antholine
View Affiliations

Affiliations: University of Wisconsin Paul P. Carbone Comprehensive Cancer Center, University of Wisconsin-Madison, Madison, WI 53792-5669, USA, University of Wisconsin Paul P. Carbone Comprehensive Cancer Center, University of Wisconsin-Madison, Madison, WI 53792-5669, USA, Department of Biostatistics, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USA, The Ohio State University Comprehensive Cancer Center, Columbus, OH 43210, USA, Department of Medicine, Medical College of Wisconsin, Milwaukee, WI 53226, USA, Department of Biophysics, Medical College of Wisconsin, Milwaukee, WI 53226, USA
Published online on: November 30, 2010 https://doi.org/10.3892/etm.2010.165
Pages: 119-123

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

3-Aminopyridine-2-carboxaldehyde thiosemicarbazone (3-AP) is a metal chelator that potently inhibits the enzyme ribonucleotide reductase (RR), which plays a key role in cell division and tumor progression. A subunit of RR has a non-heme iron and a tyrosine-free radical, which are required for the enzymatic reduction of ribonucleotides to deoxyribonucleotides. The objective of the present study was to determine whether 3-AP affects its targeted action by measuring electron paramagnetic resonance (EPR) signals formed either directly or indirectly from low molecular weight ferric-3-AP chelates. Peripheral blood lymphocytes were collected from patients with refractory solid tumors at baseline and at 2, 4.5 and 22 h after 3-AP administration. Using EPR spectra, our study identified signals from high-spin Fe-transferrin, high-spin heme and low-spin iron or copper ions. An increase in the Fe-transferrin signal was observed, suggesting blockage of Fe uptake. It is hypothesized that formation of reactive oxygen species by FeT2 or CuT damages the transferrin or the transferrin receptor. An increase in the heme signal was also observed, which was a probable source of cytochrome c release from the mitochondria and potential apoptosis. In addition, increased levels of Fe and Cu were identified. These results, which were consistent with our previous study validating 3-AP-mediated signals by EPR, provide valuable insights into the in vivo mechanism of action of 3-AP.

View Figures

View References

1.	Yu Y, Wong J, Lovejoy D, Kalinowski D and Richardson D: Chelators at the cancer coalface: desferrioxamine to Triapine and beyond. Clin Cancer Res. 12:6876–6883. 2006. View Article : Google Scholar : PubMed/NCBI
2.	Nordlund P, Sjöberg B and Eklund H: Three-dimensional structure of the free radical protein of ribonucleotide reductase. Nature. 345:593–598. 1990. View Article : Google Scholar : PubMed/NCBI
3.	Qiu W, Zhou B, Darwish D, Shao J and Yen Y: Characterization of enzymatic properties of human ribonucleotide reductase holoenzyme reconstituted in vitro from hRRM1, hRRM2, and p53R2 subunits. Biochem Biophys Res Commun. 340:428–434. 2006. View Article : Google Scholar : PubMed/NCBI
4.	Cooper C, Lynagh G, Hoyes K, Hider R, Cammack R and Porter J: The relationship of intracellular iron chelation to the inhibition and regeneration of human ribonucleotide reductase. J Biol Chem. 271:20291–20299. 1996. View Article : Google Scholar : PubMed/NCBI
5.	Shao J, Zhou B, Di Bilio A, et al: A Ferrous-Triapine complex mediates formation of reactive oxygen species that inactivate human ribonucleotide reductase. Mol Cancer Ther. 5:586–592. 2006. View Article : Google Scholar : PubMed/NCBI
6.	Kolesar J, Schelman W, Geiger P, et al: Electron paramagnetic resonance study of peripheral blood mononuclear cells from patients with refractory solid tumors treated with Triapine. J Inorg Biochem. 102:693–698. 2008. View Article : Google Scholar
7.	Petering DH and Antholine WE: Reviews in Biochemical Toxicology. 9. Hodgson E, Bend JR and Philpot RM: Elsevier; New York: 1988
8.	Richardson D, Sharpe P, Lovejoy D, et al: Dipyridyl thiosemicarbazone chelators with potent and selective antitumor activity form iron complexes with redox activity. J Med Chem. 49:6510–6521. 2006. View Article : Google Scholar : PubMed/NCBI
9.	Whitnall M, Howard J, Ponka P and Richardson D: A class of iron chelators with a wide spectrum of potent antitumor activity that overcomes resistance to chemotherapeutics. Proc Natl Acad Sci USA. 103:14901–14906. 2006. View Article : Google Scholar : PubMed/NCBI
10.	Chaston T, Lovejoy D, Watts R and Richardson D: 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. 2003.
11.	Narasimhan J, Antholine W, Chitambar C and Petering D: Inhibition of iron uptake in HL60 cells by 2-formylpyridine monothiosemicarbazonato Cu(II). Arch Biochem Biophys. 289:393–398. 1991. View Article : Google Scholar : PubMed/NCBI
12.	Martinou J, Desagher S and Antonsson B: Cytochrome c release from mitochondria: all or nothing. Nat Cell Biol. 2:E41–E43. 2000. View Article : Google Scholar : PubMed/NCBI
13.	DeConti R, Toftness B, Agrawal K, et al: Clinical and pharmacological studies wit h 5-hydroxy-2-formylpyridine thiosemicarbazone. Cancer Res. 32:1455–1462. 1972.
14.	Sartorelli A: Effect of chelating agents upon the synthesis of nucleic acids and protein: inhibition of DNA synthesis by 1-formylisoquinoline thiosemicarbazone. Biochem Biophys Res Commun. 27:26–32. 1967. View Article : Google Scholar
15.	Sartorelli A, Agrawal K and Moore E: Mechanism of inhibition of ribonucleoside diphosphate reductase by a-(N)-heterocyclic aldehyde thiosemicarbazones. Biochem Pharmacol. 20:3119–3123. 1971. View Article : Google Scholar : PubMed/NCBI
16.	Zhu L, Zhou B, Chen X, Jiang H, Shao J and Yen Y: Inhibitory mechanisms of heterocyclic carboxaldehyde thiosemicabazones for two forms of human ribonucleotide reductase. Biochem Pharmacol. 78:1178–1185. 2009. View Article : Google Scholar
17.	Kowol C, Trondl R, Arion V, Jakupec M, Lichtscheidl I and Keppler B: Fluorescence properties and cellular distribution of the investigational anticancer drug Triapine (3-aminopyridine-2-carboxaldehyde thiosemicarbazone) and its zinc(II) complex. Dalton Trans. 39:704–706. 2010. View Article : Google Scholar
18.	Attia S, Kolesar J, Mahoney M, et al: A phase 2 consortium (P2C) trial of 3-aminopyridine-2-carboxaldehyde thiosemicarbazone (3-AP) for advanced adenocarcinoma of the pancreas. Invest New Drugs. 26:369–379. 2008. View Article : Google Scholar : PubMed/NCBI
19.	Choi B, Alberti D, Schelman W, et al: The maximum tolerated dose and biologic effects of 3-aminopyridine-2-carboxaldehyde thiosemicarbazone (3-AP) in combination with irinotecan for patients with refractory solid tumors. Cancer Chemother Pharmacol. 66:973–980. 2010. View Article : Google Scholar : PubMed/NCBI
20.	Schelman W, Morgan-Meadows S, Marnocha R, et al: A phase I study of Triapine in combination with doxorubicin in patients with advanced solid tumors. Cancer Chemother Pharmacol. 63:1147–1156. 2009. View Article : Google Scholar : PubMed/NCBI
21.	Nutting C, van Herpen C, Miah A, et al: Phase II study of 3-AP Triapine in patients with recurrent or metastatic head and neck squamous cell carcinoma. Ann Oncol. 20:1275–1279. 2009. View Article : Google Scholar : PubMed/NCBI
22.	Traynor A, Lee J, Bayer G, et al: A phase II trial of Triapine (NSC# 663249) and gemcitabine as second line treatment of advanced non-small cell lung cancer: Eastern Cooperative Oncology Group Study 1503. Invest New Drugs. 28:91–97. 2010.
23.	Kunos C, Chiu S, Pink J and Kinsella T: Modulating radiation resistance by inhibiting ribonucleotide reductase in cancers with virally or mutationally silenced p53 protein. Radiat Res. 172:666–676. 2009. View Article : Google Scholar : PubMed/NCBI
24.	Antholine W, Knight J, Whelan H and Petering D: Studies of the reaction of 2-formylpyridine thiosemicarbazone and its iron and copper complexes with biological systems. Mol Pharmacol. 13:89–98. 1977.PubMed/NCBI
25.	Saryan L, Mailer K, Krishnamurti C, Antholine W and Petering D: Interaction of 2-formylpyridine thiosemicarbazonato copper (II) with Ehrlich ascites tumor cells. Biochem Pharmacol. 30:1595–1604. 1981. View Article : Google Scholar : PubMed/NCBI
26.	Alvero A, Chen W, Sartorelli A, Schwartz P, Rutherford T and Mor G: Triapine (3-aminopyridine-2-carboxaldehyde thiosemicarbazone) induces apoptosis in ovarian cancer cells. J Soc Gynecol Investig. 13:145–152. 2006. View Article : Google Scholar : PubMed/NCBI