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Biological properties of 4-methyl-2,7-diamino-5,10-diphenyl-4,9-diazapyrenium hydrogensulfate (ADAP)

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Abstract

Objective

4-Methyl-2,7-diamino-5,10-diphenyl-4,9-diazapyrenium hydrogensulfate (ADAP) is a potential antitumor compound because of its DNA and RNA intercalating ability. In this study, cellular uptake, intracellular distribution as well as mechanism of action, antitumor activity in vitro and toxicity in vivo of ADAP were investigated.

Methods

Based on the fluorescence properties of ADAP, its entry and distribution into live cells were analyzed by fluorescence microscopy. The in vitro antiproliferative activity was determined using MTT test. For screening of topoisomerase II-targeted effects of ADAP, the cell-free assay and immunoband depletion assay were used. Expression of the genes c-mos, c-N-ras, c-Ki-ras, c-H-ras, p53 and caspase 3 in Caco-2 cells treated with ADAP was examined by RT-PCR. Toxicity in vivo was determined using C3HHf/Bu Zgr/Hr mice treated by single or multiple doses of ADAP at a concentration of 25 mg/kg.

Results

ADAP in μM concentrations entered into MIAPaCa-2 cell’s cytoplasm in 5 min and into nuclei in 60 min after administration. Intracellular distribution of ADAP depended on the period of treatment time. ADAP (0.1–100 μM) strongly inhibited the growth of both mouse (FsaR, SCCVII) and human tumor cells (HeLa, Caco-2, HT-29, MIAPaCa-2, HBL, HEp-2, SW620, MCF-7) compared to its weak cytotoxicity on controls and normal cells (WI38). Results of both topoisomerase II assays showed that ADAP is not a topoisomerase II poison. Expression of investigated genes was dependent on the incubation time, except for p53 and c-H-ras. Morphological changes in tissues and organs of mice were not observed. Results of patohistological analysis have been confirmed by hematological and clinical-chemical analysis of blood of treated and non-treated animals.

Conclusion

ADAP is a strongly bioactive compound with antitumor potential in vitro. The antitumor potential in vivo remains to be identified.

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Acknowledgments

This project was supported by the Ministry of Science, Education and Sport, Republic of Croatia, grants 0127111 and 219-0982914-2176. We are grateful to Prof. Dr. Mirjana Knežević-Kostović and M.Sc. Neda Cetina for their kind support.

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Authors and Affiliations

  1. Department of Nuclear Medicine and Pathophysiology, Clinical Hospital Osijek, Osijek, Croatia

    Saška Marczi & Ljubica Glavaš-Obrovac

  2. Department of Chemistry, Biochemistry and Clinical Chemistry, School of Medicine Osijek, Huttlerova 4, 31000, Osijek, Croatia

    Ljubica Glavaš-Obrovac

  3. Department of Histology and Embriology, School of Medicine Osijek, Osijek, Croatia

    Tatjana Belovari

  4. Division of Molecular Medicine, Ruđer Bošković Institute, Zagreb, Croatia

    Ranko Stojković & Siniša Ivanković

  5. Department of Clinical Biochemistry, Clinical Hospital Osijek, Osijek, Croatia

    Vatroslav Šerić

  6. Division of Organic Chemistry and Biochemistry, Ruđer Bošković Institute, Zagreb, Croatia

    Ivo Piantanida & Mladen Žinić

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  1. Saška Marczi
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Correspondence to Ljubica Glavaš-Obrovac.

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Marczi, S., Glavaš-Obrovac, L., Belovari, T. et al. Biological properties of 4-methyl-2,7-diamino-5,10-diphenyl-4,9-diazapyrenium hydrogensulfate (ADAP). Cancer Chemother Pharmacol 62, 595–604 (2008). https://doi.org/10.1007/s00280-007-0643-0

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