Skip to main content
SpringerLink
Log in

Novel isosteric charge-deficient spermine analogue—1,12-diamino-3,6,9-triazadodecane: synthesis, pK a measurement and biological activity

  • Original Article
  • Published:
Amino Acids Aims and scope Submit manuscript

Abstract

Ionic interactions are essential for the biological functions of the polyamines spermidine and spermine in mammalian physiology. Here, we describe a simple gram scale method to prepare 1,12-diamino-3,6,9-triazadodecane (SpmTrien), an isosteric charge-deficient spermine analogue. The protonation sites of SpmTrien were determined at pH range of 2.2–11.0 using two-dimensional 1H-15N NMR spectroscopy, which proved to be more feasible than conventional methods. The macroscopic pK a values of SpmTrien (3.3, 6.3, 8.5, 9.5 and 10.3) are significantly lower than those of 1,12-diamino-4,9-diazadodecane (spermine). The effects of SpmTrien and its parent molecule, 1,8-diamino-3,6-diazaoctane (Trien), on cell growth and polyamine metabolism were investigated in DU145 prostate carcinoma cells. SpmTrien downregulated the biosynthetic enzymes ornithine decarboxylase (ODC) and S-adenosyl-l-methionine decarboxylase and decreased intracellular polyamine levels, whereas the effects of Trien alone were minor. Interestingly, both SpmTrien and Trien were able to partially overcome growth arrest induced by an ODC inhibitor, α-difluoromethylornithine, indicating that they are able to mimic some functions of the natural polyamines. Thus, SpmTrien is a novel tool to influence polyamine interaction sites at the molecular level and offers a new means to study the contribution of the protonation of spermine amino group(s) in the regulation of polyamine-dependent biological processes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Scheme 1
Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Abbreviations

AdoMetDC:

S-adenosyl-L-methionine decarboxylase

AG:

Aminoguanidine

DENSpm:

N1,N11-diethylnorspermine

DFMO:

α-Difluoromethylornithine (2,5-diamino-2-(difluoromethyl)pentanoic acid)

HMBC:

Heteronuclear multiple bond correlation

HSQC:

Heteronuclear single quantum correlation

ODC:

Ornithine decarboxylase

Pu:

Putrescine (1,4-diaminobutane)

Spm:

Spermine (1,12-diamino-4,9-diazadodecane)

SpmTrien:

(1,12-Diamino-3,6,9-triazadodecane)

Spd:

Spermidine (1,8-diamino-4-azaoctane)

SSAT:

Spermidine/spermine N 1-acetyltransferase

Trien:

Triethylenetetramine (1,8-diamino-3,6-diazaoctane)

TSP:

Sodium 3-trimethylsilyl[2,2,3,3-d 4]propionate

References

  • Agostinelli E, De Matteis G, Mondovì B, Morpurgo L (1998) Reconstitution of Cu2+-depleted bovine serum amine oxidase with Co2+. Biochem J 330:383–387

    CAS  PubMed  Google Scholar 

  • Baillon J, Mamont PS, Wagner J, Gerhart F, Lux P (1988) Fluorinated analogues of spermidine as substrates of spermine synthase. Eur J Biochem 176:237–242

    Article  CAS  PubMed  Google Scholar 

  • Basu HS, Schwietert HC, Feuerstein BG, Marton LJ (1990) Effects of variation in the structure of spermine on the association with DNA and the induction of DNA conformational changes. Biochem J 269:329–334

    CAS  PubMed  Google Scholar 

  • Bencini A, Bianchi A, Garcia-España E, Micheloni M, Ramirez JA (1999) Proton coordination by polyamine compounds in aqueous solution. Coord Chem Rev 188:97–156

    Article  CAS  Google Scholar 

  • Bergeron RJ, Neims AH, McManis JS, Hawthorne TR, Vinson JR, Bortell R, Ingeno MJ (1988) Synthetic polyamine analogues as antineoplastics. J Med Chem 31:1183–1190

    Article  CAS  PubMed  Google Scholar 

  • Bergeron RJ, McManis JS, Weimar WR, Schreier KM, Gao F, Wu Q, Ortiz-Ocasio J, Luchetta GR, Porter C, Vinson JR (1995) The role of charge in polyamine analogue recognition. J Med Chem 38:2278–2285

    Article  PubMed  Google Scholar 

  • Bernacki RJ, Bergeron RJ, Porter CW (1992) Antitumor activity of N,N′-bis(ethyl)spermine homologues against human MALME-3 melanoma xenografts. Cancer Res 52:2424–2430

    CAS  PubMed  Google Scholar 

  • Casero RA Jr, Marton LJ (2007) Targeting polyamine metabolism and function in cancer and other hyperproliferative diseases. Nat Rev Drug Discov 6:373–390

    Article  CAS  PubMed  Google Scholar 

  • Casero RA, Woster PM (2001) Terminally alkylated polyamine analogues as chemotherapeutic agents. J Med Chem 44:1–26

    Article  CAS  PubMed  Google Scholar 

  • Cooper GJS, Phillips ARJ, Choong SY et al (2004) Regeneration of the heart in diabetes by selective copper chelation. Diabetes 53:2501–2508

    Article  CAS  PubMed  Google Scholar 

  • Dixon HBF, Gibbs K, Walshe JM (1972) Preparation of triethylenetetramine dihydrochloride for the treatment of Wilson’s disease. Lancet 1:853

    Article  CAS  PubMed  Google Scholar 

  • Eloranta TO, Khomutov AR, Khomutov RM, Hyvönen T (1990) Aminooxy analogues of spermidine as inhibitors of spermine synthase and substrates of hepatic polyamine acetylating activity. J Biochem (Tokyo) 108:593–598

    CAS  Google Scholar 

  • Frassineti C, Ghelli S, Gans P, Sabatini A, Moruzzi MS, Vacca A (1995) Nuclear magnetic resonance as a tool for determining protonation constants of natural polyprotic bases in solution. Anal Biochem 231:374–382

    Article  CAS  PubMed  Google Scholar 

  • Hyvönen T, Keinänen TA, Khomutov AR, Khomutov RM, Eloranta TO (1992) Monitoring of the uptake and metabolism of aminooxy analogues of polyamines in cultured cells by high-performance liquid chromatography. J Chromatogr 574:17–21

    Google Scholar 

  • Hyvönen T, Keinänen TA, Khomutov AR, Khomutov RM, Eloranta TO (1995) Aminooxy analogues of spermidine evidence the divergent roles of the charged amino nitrogens in the cellular physiology of spermidine. Life Sci 56:349–360

    Article  PubMed  Google Scholar 

  • Hyvönen MT, Howard M, Grigorenko N, Khomutov AR, Vepsäläinen J, Alhonen L, Jänne J, Keinänen TA (2009) Divergent regulation of the key metabolic enzymes of polyamine metabolism by chiral α-methylated polyamine analogs. Biochem J 422:321–328

    Article  PubMed  Google Scholar 

  • Jänne J, Williams-Ashman HG (1971) On the purification of l-ornithine decarboxylase from rat prostate and effects of thiol compounds on the enzyme. J Biol Chem 246:1725–1732

    PubMed  Google Scholar 

  • Jones M, Singh P, Zimmerman L, Gomez M, Albina L, Domingo J (1995) Effects of some chelating agents on urinary copper excretion by the rat. Chem Res Toxicol 8:942–948

    Article  CAS  PubMed  Google Scholar 

  • Khomutov AR, Vepsäläinen J, Shvetsov AS, Hyvonen T, Keinanen TA, Pustobaev VN, Eloranta TO, Khomutov RM (1996) Synthesis of hydroxylamine analogues of polyamines. Tetrahedron 52:13751–13766

    Article  CAS  Google Scholar 

  • Khomutov AR, Simonian AR, Vepsäläinen J, Keinänen TA, Alhonen L, Jänne J (2005) New oxaanalogues of spermine. Russ J Bioorganic Chem 31:206–212

    CAS  Google Scholar 

  • Khomutov AR, Grigorenko NA, Skuridin SG (2007) Novel approach to design isosteric charge-deficient analogue of spermine and its biochemically important derivatives. Biochem Soc Trans 35:369–373

    Article  CAS  PubMed  Google Scholar 

  • Kobayashi M, Fujisaki H, Sugawara M, Iseki K, Miyazaki K (1999) The presence of an Na+/spermine antiporter in the rat renal brush-border membrane. J Pharm Pharmacol 51:279–284

    Article  CAS  PubMed  Google Scholar 

  • Laatikainen R, Niemitz M, Weber U, Sundelin J, Hassinen T, Vepsäläinen J (1996) General strategies for total-lineshape-type spectral analysis of NMR spectra using integral-transform iterator. J Magn Reson Ser A 120:1–10

    Article  CAS  Google Scholar 

  • Lin PKT, Maguire NM, Brown DM (1994) Synthesis of novel oxa-isosteres of spermidine and spermine. Tetrahedron Lett 35:3605–3608

    Article  CAS  Google Scholar 

  • Lu J, Chan YK, Gamble GD, Poppitt SD, Othman AA, Cooper GJ (2007) Triethylenetetramine and metabolites: levels in relation to copper and zinc excretion in urine of healthy volunteers and type 2 diabetic patients. Drug Metab Dispos 35:221–227

    Article  CAS  PubMed  Google Scholar 

  • Nocera S, Marcocci L, Pietrangeli P, Mondovì B (2003) New perspectives on the role of amine oxidases in physiopathology. Amino Acids 24:13–17

    CAS  PubMed  Google Scholar 

  • Onufriev A, Case DA, Ullmann GM (2001) A novel view of pH titration in biomolecules. Biochemistry 40:3413–3419

    Article  CAS  PubMed  Google Scholar 

  • Ramirez FJ, Thomas TJ, Antony T, Ruiz-Chica J, Thomas T (2002) Effects of aminooxy analogues of biogenic polyamines on aggregation and stability of calf thymus DNA. Biopolymers 65:148–157

    Article  CAS  PubMed  Google Scholar 

  • Schilsky ML (2001) Treatment of Wilson’s disease: what are the relative roles of penicillamine, trientine, and zinc supplementation? Curr Gastroenterol Rep 3:54–59

    Article  CAS  PubMed  Google Scholar 

  • Schwarzenbach G (1950) Metallkomplexe mit Polyaminen III: Mit Triäthylen-tetramin = “trien”. Helv Chem Acta 33:974–985

    Article  CAS  Google Scholar 

  • Seiler N (2004) Catabolism of polyamines. Amino Acids 26:217–233

    CAS  PubMed  Google Scholar 

  • Szakacs Z, Kraszni M, Noszal B (2004) Determination of microscopic acid-base parameters from NMR-pH titration. Anal Bioanal Chem 378:1428–1448

    Article  CAS  PubMed  Google Scholar 

  • Turchanowa L, Shvetsov AS, Demin AV, Khomutov AR, Wallace HM, Stein J, Milovic V (2002) Insufficiently changed isosteric analogue of spermine: interaction with polyamine uptake, and effect on Caco-2 cell growth. Biochem Pharmacol 64:649–655

    Article  CAS  PubMed  Google Scholar 

  • Ullmann GM (2003) Relations between protonation constants and titration curves in polyprotic acids: a critical view. J Phys Chem B 107:1263–1271

    Article  CAS  Google Scholar 

  • Wallace HM, Fraser AV (2004) Inhibitors of polyamine metabolism: review article. Amino Acids 26:353–365

    Article  CAS  PubMed  Google Scholar 

  • Wallace HM, Niiranen K (2007) Polyamine analogues–an update. Amino Acids 33:261–265

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We would like to thank Ms. Maritta Salminkoski, Ms. Tuula Reponen, Ms. Anne Karppinen, Ms. Arja Korhonen for their skilful technical assistance and Ph.D. Elina Jarho for her assistance with the automatic titrator. This work was supported by Academy of Finland (project nos. 124185 and 128702), Russian Foundation for Basic Research (project no. 09-04-01272), and the program Molecular and Cell Biology of the Presidium of the Russian Academy of Sciences.

Author information

Authors and Affiliations

  1. Laboratory of Chemistry, Department of Biosciences, University of Kuopio, Kuopio, Finland

    Janne Weisell, Jouko Vepsäläinen & Pasi Soininen

  2. Department of Biotechnology and Molecular Medicine, A. I. Virtanen Institute, Biocenter Kuopio, University of Kuopio, Kuopio, Finland

    Mervi T. Hyvönen, Leena Alhonen & Tuomo A. Keinänen

  3. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia

    Alex R. Khomutov

Authors
  1. Janne Weisell
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mervi T. Hyvönen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jouko Vepsäläinen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Leena Alhonen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tuomo A. Keinänen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Alex R. Khomutov
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Pasi Soininen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Janne Weisell.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Weisell, J., Hyvönen, M.T., Vepsäläinen, J. et al. Novel isosteric charge-deficient spermine analogue—1,12-diamino-3,6,9-triazadodecane: synthesis, pK a measurement and biological activity. Amino Acids 38, 501–507 (2010). https://doi.org/10.1007/s00726-009-0409-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00726-009-0409-6

Keywords

Search

Navigation