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Iron containing anti-tumoral agents: unexpected apoptosis-inducing activity of a ferrocene amino acid derivative

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Abstract

Purpose

Due to the severe problems accompanied with multiple drug resistance (MDR), agents that can induce apoptosis independently of death-suppressing proteins are required. Here, we show that the ferrocene derivative HUNI 068 is active against cancer cells and overcomes different mechanisms of multiple drug resistance (MDR).

Methods

Proliferation inhibition was determined by using a CASY®CellCounter. DNA fragmentation assay and annexin-V/PI binding assays measured apoptosis, and necrosis was excluded by LDH-release assay. Drug-resistant cell lines were generated to test the ability to overcome MDR. By real-time PCR, alterations in gene expression of treated cells were analyzed. The apoptosis pathway was investigated by immunoblotting and measurement of mitochondrial membrane permeability transition.

Results

HUNI 068 leads to proliferation inhibition and apoptosis mediation, but only minimal necrosis induction. Healthy leukocytes seem to be less affected than cancer cells. The compound overcomes drug resistance to vincristine and daunorubicin. Independence of p-glycoprotein and Bcl-2 overexpression is probable, and upregulation of the anti-Bcl-2 protein harakiri was seen. Combined treatment with vincristine leads to synergistic effects. In different primary tumor cells, HUNI 068 achieved acceptable effects where tolerance to some conventional drugs was shown. Induction of apoptosis is FADD-independent, but associated with a reduced mitochondrial membrane potential and activation of caspase-9, indicating the intrinsic apoptosis pathway via mitochondria.

Conclusions

HUNI 068 is a promising new compound with activity even against MDR tumor cells. Further investigations into the class of ferrocene-derived agents might reveal compounds with improved activity for a more specific and safe anti-cancer therapy.

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References

  • Allardyce CS, Dorcier A, Scolaro C, Dyson PJ (2005) Development of organometallic (organo-transition metal) pharmaceuticals. Appl Organomet Chem 19:1–10

    Article  CAS  Google Scholar 

  • Bene MC, Castoldi G, Knapp W, Ludwig WD, Matutes E, Orfao A, van’t Veer MB (1995) Proposals for the immunological classification of acute leukemias. European Group for the immunological characterization of leukemias (EGIL). Leukemia 9:1783–1786

    PubMed  CAS  Google Scholar 

  • Bene MC, Bernier M, Castoldi G, Faure GC, Knapp W, Ludwig WD, Matutes E, Orfao A, van’t Veer M (1999) Impact of immunophenotyping on management of acute leukemias. Haematologica 84:1024–1034

    PubMed  CAS  Google Scholar 

  • Brozek J, Bryl E, Ploszynska A, Balcerska A, Witkowski JM (2009) P-glycoprotein activity predicts outcome in childhood acute lymphoblastic leukemia. J Pediatr Hematol Oncol 31:493–499

    Article  PubMed  CAS  Google Scholar 

  • Essmann F, Wieder T, Otto A, Muller EC, Dörken B, Daniel PT (2000) GDP dissociation inhibitor D4-GDI (Rho-GDI 2), but not the homologous rho-GDI 1, is cleaved by caspase-3 during drug-induced apoptosis. Biochem J 346(Pt 3):777–783

    Article  PubMed  CAS  Google Scholar 

  • Fadok VA, Xue D, Henson P (2001a) If phosphatidylserine is the death knell, a new phosphatidylserine-specific receptor is the bellringer. Cell Death Differ 8:582–587

    Article  PubMed  CAS  Google Scholar 

  • Fadok VA, de Cathelineau A, Daleke DL, Henson PM, Bratton DL (2001b) Loss of phospholipid asymmetry and surface exposure of phosphatidylserine is required for phagocytosis of apoptotic cells by macrophages and fibroblasts. J Biol Chem 276:1071–1077

    Article  PubMed  CAS  Google Scholar 

  • Gaynon PS, Qu RP, Chappell RJ, Willoughby ML, Tubergen DG, Steinherz PG, Trigg ME (1998) Survival after relapse in childhood acute lymphoblastic leukemia: impact of site and time to first relapse—the children’s cancer group experience. Cancer 82:1387–1395

    Article  PubMed  CAS  Google Scholar 

  • Hunold A, Neundorf I, James P, Neudörfl J, Schmalz HG (2009) Stereoselective synthesis of new ferrocene-derived amino acid building blocks. Eur J Org Chem 4429–4440

  • Inohara N, Ding L, Chen S, Nunez G (1997) Harakiri, a novel regulator of cell death, encodes a protein that activates apoptosis and interacts selectively with survival-promoting proteins Bcl-2 and Bcl-X(L). EMBO J 16:1686–1694

    Article  PubMed  CAS  Google Scholar 

  • James P, Neudorfl J, Eissmann M, Jesse P, Prokop A, Schmalz HG (2006) Enantioselective synthesis of ferrocenyl nucleoside analogues with apoptosis-inducing activity. Org Lett 8:2763–2766

    Article  PubMed  CAS  Google Scholar 

  • Jaouen G (2005) Bioorganometallics: biomolecules, labeling, medicine. Viley-VCH, New York

    Google Scholar 

  • Jenkins CE, Swiatoniowski A, Power MR, Lin TJ (2006) Pseudomonas aeruginosa-induced human mast cell apoptosis is associated with up-regulation of endogenous Bcl-xS and down-regulation of Bcl-xL. J Immunol 177:8000–8007

    PubMed  CAS  Google Scholar 

  • Kealy T, Pauson P (1951) A new type of organo-iron compound. Nature 168:1039–1040

    Article  CAS  Google Scholar 

  • Kopf-Maier P, Erkenswick P (1984) Teratogenicity and embryotoxicity of titanocene dichloride in mice. Toxicology 33:171–181

    Article  PubMed  CAS  Google Scholar 

  • Krishna R, Mayer LD (2000) Multidrug resistance (MDR) in cancer. Mechanisms, reversal using modulators of MDR and the role of MDR modulators in influencing the pharmacokinetics of anticancer drugs. Eur J Pharm Sci 11:265–283

    Article  PubMed  CAS  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  PubMed  CAS  Google Scholar 

  • Lambert IH, Hoffmann EK, Jorgensen F (1989) Membrane potential, anion and cation conductances in Ehrlich ascites tumor cells. J Membr Biol 111:113–131

    Article  PubMed  CAS  Google Scholar 

  • Nagata S (1997) Apoptosis by death factor. Cell 88:355–365

    Article  PubMed  CAS  Google Scholar 

  • Nguyen A, Vessières A, Hillard E, Top S, Pigeon P, Jaouen G (2007) Ferrocifens and ferrocifenols as new potential weapons against breast cancer. Chimia 61:716–724

    Article  CAS  Google Scholar 

  • Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C (1991) A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 139:271–279

    Article  PubMed  CAS  Google Scholar 

  • Prokop A, Wieder T, Sturm I, Essmann F, Seeger K, Wuchter C, Ludwig WD, Henze G, Dörken B, Daniel PT (2000) Relapse in childhood acute lymphoblastic leukemia is associated with a decrease of the Bax/Bcl-2 ratio and loss of spontaneous caspase-3 processing in vivo. Leukemia 14:1606–1613

    Article  PubMed  CAS  Google Scholar 

  • Prokop A, Wrasidlo W, Lode H, Herold R, Lang F, Henze G, Dorken B, Wieder T, Daniel PT (2003) Induction of apoptosis by enediyne antibiotic calicheamicin thetaII proceeds through a caspase-mediated mitochondrial amplification loop in an entirely Bax-dependent manner. Oncogene 22:9107–9120

    Article  PubMed  CAS  Google Scholar 

  • Reers M, Smiley ST, Mottola-Hartshorn C, Chen A, Lin M, Chen LB (1995) Mitochondrial membrane potential monitored by JC-1 dye. Methods Enzymol 260:406–417

    Article  PubMed  CAS  Google Scholar 

  • Schatzschneider U, Metzler-Nolte N (2006) New principles in medicinal organometallic chemistry. Angew Chem Int Ed Engl 45:1504–1507

    Article  PubMed  CAS  Google Scholar 

  • Schlawe D, Majdalani A, Velcicky J, Hessler E, Wieder T, Prokop A, Schmalz HG (2004) Iron-containing nucleoside analogues with pronounced apoptosis-inducing activity. Angew Chem Int Ed Engl 43:1731–1734

    Article  PubMed  CAS  Google Scholar 

  • Schlegel RA, Williamson P (2001) Phosphatidylserine, a death knell. Cell Death Differ 8:551–563

    Article  PubMed  CAS  Google Scholar 

  • Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchoninic acid. Anal Biochem 150:76–85

    Article  PubMed  CAS  Google Scholar 

  • Srinivas G, Kusumakumary P, Nair MK, Panicker KR, Pillai MR (2000) Mutant p53 protein, Bcl-2/Bax ratios and apoptosis in paediatric acute lymphoblastic leukaemia. J Cancer Res Clin Oncol 126:62–67

    Article  PubMed  CAS  Google Scholar 

  • Tillner J, Winckler T, Dingermann T (1994) A simple cytotoxicity assay using the eucaryotic microorganism Dictyostelium discoideum. Pharmazie 49:759–761

    PubMed  CAS  Google Scholar 

  • Top S, Vessieres A, Leclercq G, Quivy J, Tang J, Vaissermann J, Huche M, Jaouen G (2003) Synthesis, biochemical properties and molecular modelling studies of organometallic specific estrogen receptor modulators (SERMs), the ferrocifens and hydroxyferrocifens: evidence for an antiproliferative effect of hydroxyferrocifens on both hormone-dependent and hormone-independent breast cancer cell lines. Chem Eur J 9:5223–5236

    Article  CAS  Google Scholar 

  • Uderzo C, Conter V, Dini G, Locatelli F, Miniero R, Tamaro P (2001) Treatment of childhood acute lymphoblastic leukemia after the first relapse: curative strategies. Haematologica 86:1–7

    PubMed  CAS  Google Scholar 

  • Van Cruchten S, Van Den Broeck W (2002) Morphological and biochemical aspects of apoptosis, oncosis and necrosis. Anat Histol Embryol 31:214–223

    Article  PubMed  Google Scholar 

  • Vermes I, Haanen C, Steffens-Nakken H, Reutelingsperger C (1995) A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods 184:39–51

    Article  PubMed  CAS  Google Scholar 

  • Viotte M, Gautheron B, Kubicki MM, Nifant’ev IE, Fricker SP (1995) Synthesis, structure of nitrogen-containing phosphinogold(I) ferrocenes. In vitro Activity against bladder and colon carcinoma cell lines. Met Based Drugs 2:311–326

    Article  PubMed  CAS  Google Scholar 

  • Voisard R, Dartsch PC, Seitzer U, Roth D, Kochs M, Hombach V (1991) Cell culture as a prescreening system for drug prevention of restenosis? Vasa Suppl 33:140–141

    PubMed  CAS  Google Scholar 

  • von Haefen C, Wieder T, Essmann F, Schulze-Osthoff K, Dorken B, Daniel PT (2003) Paclitaxel-induced apoptosis in BJAB cells proceeds via a death receptor-independent, caspases-3–8-driven mitochondrial amplification loop. Oncogene 22:2236–2247

    Article  Google Scholar 

  • Wieder T, Geilen CC, Wieprecht M, Becker A, Orfanos CE (1994) Identification of a putative membrane-interacting domain of CTP: phosphocholine cytidylyltransferase from rat liver. FEBS Lett 345:207–210

    Article  PubMed  CAS  Google Scholar 

  • Wieder T, Orfanos CE, Geilen CC (1998) Induction of ceramide-mediated apoptosis by the anticancer phospholipid analog, hexadecylphosphocholine. J Biol Chem 273:11025–11031

    Article  PubMed  CAS  Google Scholar 

  • Wieder T, Essmann F, Prokop A, Schmelz K, Schulze-Osthoff K, Beyaert R, Dörken B, Daniel PT (2001) Activation of caspase-8 in drug-induced apoptosis of B-lymphoid cells is independent of CD95/Fas receptor-ligand interaction and occurs downstream of caspase-3. Blood 97:1378–1387

    Article  PubMed  CAS  Google Scholar 

  • Wilkinson G (1975) The iron sandwich. A recollection of the first four months. J Organometal Chem 100:273–278

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by Deutsche Forschungsgemeinschaft (DFG, Germany; FOR 630), Kinderleben e.V. (Berlin, Germany) and Dr. Kleist-Stiftung (Berlin, Germany). The excellent technical assistance of Corazon Frias, Soo-Young Lee, Liliane Abodo and Ulrike Schmitt is gratefully acknowledged.

Conflicts of interest statement

The authors declare that they have no competing interests.

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Authors and Affiliations

  1. Department of Pediatric Oncology/Hematology, University Medical Center Charité, Campus Virchow, Augustenburger Platz 1, 13353, Berlin, Germany

    Benjamin Kater, Lisa Kater, Birgit Bonitzki, Patrick Jesse & Aram Prokop

  2. Department of Organic Chemistry, University Cologne, Greinstraße 4, 50939, Cologne, Germany

    Andrea Hunold & Hans-G. Schmalz

  3. Department of Pediatric Hematology/Oncology, Children’s Hospital Cologne, Amsterdamerstraße 59, 50735, Cologne, Germany

    Aram Prokop

  4. Knesebeckstraße 89, 10623, Berlin, Germany

    Benjamin Kater

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  1. Benjamin Kater
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  2. Andrea Hunold
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  4. Lisa Kater
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Correspondence to Benjamin Kater.

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Kater, B., Hunold, A., Schmalz, HG. et al. Iron containing anti-tumoral agents: unexpected apoptosis-inducing activity of a ferrocene amino acid derivative. J Cancer Res Clin Oncol 137, 639–649 (2011). https://doi.org/10.1007/s00432-010-0924-6

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  • DOI: https://doi.org/10.1007/s00432-010-0924-6

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