Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases

Merja R. Häkkinen, Mervi T. Hyvönen, Seppo Auriola, Robert A Casero, Jouko Vepsäläinen, Alex R. Khomutov, Leena Alhonen, Tuomo A. Keinänen

Research output: Contribution to journalArticle

Abstract

N-alkylated polyamine analogues have potential as anticancer and antiparasitic drugs. However, their metabolism in the host has remained incompletely defined thus potentially limiting their utility. Here, we have studied the degradation of three different spermine analogues N,N′-bis-(3-ethylaminopropyl)butane-1,4-diamine (DESPM), N-(3-benzyl-aminopropyl)-N′-(3-ethylamino-propyl)butane-1,4-diamine (BnEtSPM) and N,N′-bis-(3-benzylaminopropyl)butane-1,4-diamine (DBSPM) and related mono-alkylated derivatives as substrates of recombinant human polyamine oxidase (APAO) and spermine oxidase (SMO). APAO and SMO metabolized DESPM to EtSPD [Km(APAO) = 10 μM, kcat(APAO) = 1.1 s -1 and Km(SMO) = 28 μM, kcat(SMO) = 0.8 s-1, respectively], metabolized BnEtSPM to EtSPD [Km(APAO) = 0.9 μM, kcat(APAO) = 1.1 s-1 and Km(SMO) = 51 μM, kcat(SMO) = 0.4 s-1, respectively], and metabolized DBSPM to BnSPD [Km(APAO) = 5.4 μM, k cat(APAO) = 2.0 s-1and Km(SMO) = 33 μM, kcat(SMO) = 0.3 s-1, respectively]. Interestingly, mono-alkylated spermine derivatives were metabolized by APAO and SMO to SPD [EtSPM Km(APAO) =16 μM, kcat(APAO) = 1.5 s -1; Km(SMO) = 25 μM, kcat(SMO) = 8.2 s -1; BnSPM Km(APAO) = 6.0 μM, kcat(APAO) = 2.8 s-1; Km(SMO) =19 μM, kcat(SMO) = 0.8 s-1, respectively]. Surprisingly, EtSPD [Km(APAO) = 37 μM, kcat(APAO) = 0.1 s-1; Km(SMO) =48 μM, kcat(SMO) = 0.05 s-1] and BnSPD [Km(APAO) = 2.5 μM, kcat(APAO) = 3.5 s-1; Km(SMO) =60 μM, kcat(SMO) = 0.54 s-1] were metabolized to SPD by both the oxidases. Furthermore, we studied the degradation of DESPM, BnEtSPM or DBSPM in the DU145 prostate carcinoma cell line. The same major metabolites EtSPD and/or BnSPD were detected both in the culture medium and intracellularly after 48 h of culture. Moreover, EtSPM and BnSPM were detected from cell samples. Present data shows that inducible SMO parallel with APAO could playanimportant roleinpolyamine based drug action, i.e. degradation of parent drug and its metabolites, having significant impact on efficiency of these drugs, and hence for the development of novel N-alkylated polyamine analogues.

Original languageEnglish (US)
Pages (from-to)369-381
Number of pages13
JournalAmino Acids
Volume38
Issue number2
DOIs
StatePublished - Feb 2010

Fingerprint

Spermine
Metabolism
Diamines
polyamine oxidase
acetylaminophenylarsine oxide
Polyamines
Metabolites
Degradation
Pharmaceutical Preparations
Derivatives
Antiparasitic Agents

Keywords

  • Flavin-dependent amino-oxidoreductases
  • N-alkylated polyamine analogues
  • Polyamine oxidase
  • Polyamines
  • Spermine oxidase

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Organic Chemistry

Cite this

Häkkinen, M. R., Hyvönen, M. T., Auriola, S., Casero, R. A., Vepsäläinen, J., Khomutov, A. R., ... Keinänen, T. A. (2010). Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases. Amino Acids, 38(2), 369-381. https://doi.org/10.1007/s00726-009-0429-2

Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases. / Häkkinen, Merja R.; Hyvönen, Mervi T.; Auriola, Seppo; Casero, Robert A; Vepsäläinen, Jouko; Khomutov, Alex R.; Alhonen, Leena; Keinänen, Tuomo A.

In: Amino Acids, Vol. 38, No. 2, 02.2010, p. 369-381.

Research output: Contribution to journalArticle

Häkkinen, MR, Hyvönen, MT, Auriola, S, Casero, RA, Vepsäläinen, J, Khomutov, AR, Alhonen, L & Keinänen, TA 2010, 'Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases', Amino Acids, vol. 38, no. 2, pp. 369-381. https://doi.org/10.1007/s00726-009-0429-2
Häkkinen MR, Hyvönen MT, Auriola S, Casero RA, Vepsäläinen J, Khomutov AR et al. Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases. Amino Acids. 2010 Feb;38(2):369-381. https://doi.org/10.1007/s00726-009-0429-2
Häkkinen, Merja R. ; Hyvönen, Mervi T. ; Auriola, Seppo ; Casero, Robert A ; Vepsäläinen, Jouko ; Khomutov, Alex R. ; Alhonen, Leena ; Keinänen, Tuomo A. / Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases. In: Amino Acids. 2010 ; Vol. 38, No. 2. pp. 369-381.
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abstract = "N-alkylated polyamine analogues have potential as anticancer and antiparasitic drugs. However, their metabolism in the host has remained incompletely defined thus potentially limiting their utility. Here, we have studied the degradation of three different spermine analogues N,N′-bis-(3-ethylaminopropyl)butane-1,4-diamine (DESPM), N-(3-benzyl-aminopropyl)-N′-(3-ethylamino-propyl)butane-1,4-diamine (BnEtSPM) and N,N′-bis-(3-benzylaminopropyl)butane-1,4-diamine (DBSPM) and related mono-alkylated derivatives as substrates of recombinant human polyamine oxidase (APAO) and spermine oxidase (SMO). APAO and SMO metabolized DESPM to EtSPD [Km(APAO) = 10 μM, kcat(APAO) = 1.1 s -1 and Km(SMO) = 28 μM, kcat(SMO) = 0.8 s-1, respectively], metabolized BnEtSPM to EtSPD [Km(APAO) = 0.9 μM, kcat(APAO) = 1.1 s-1 and Km(SMO) = 51 μM, kcat(SMO) = 0.4 s-1, respectively], and metabolized DBSPM to BnSPD [Km(APAO) = 5.4 μM, k cat(APAO) = 2.0 s-1and Km(SMO) = 33 μM, kcat(SMO) = 0.3 s-1, respectively]. Interestingly, mono-alkylated spermine derivatives were metabolized by APAO and SMO to SPD [EtSPM Km(APAO) =16 μM, kcat(APAO) = 1.5 s -1; Km(SMO) = 25 μM, kcat(SMO) = 8.2 s -1; BnSPM Km(APAO) = 6.0 μM, kcat(APAO) = 2.8 s-1; Km(SMO) =19 μM, kcat(SMO) = 0.8 s-1, respectively]. Surprisingly, EtSPD [Km(APAO) = 37 μM, kcat(APAO) = 0.1 s-1; Km(SMO) =48 μM, kcat(SMO) = 0.05 s-1] and BnSPD [Km(APAO) = 2.5 μM, kcat(APAO) = 3.5 s-1; Km(SMO) =60 μM, kcat(SMO) = 0.54 s-1] were metabolized to SPD by both the oxidases. Furthermore, we studied the degradation of DESPM, BnEtSPM or DBSPM in the DU145 prostate carcinoma cell line. The same major metabolites EtSPD and/or BnSPD were detected both in the culture medium and intracellularly after 48 h of culture. Moreover, EtSPM and BnSPM were detected from cell samples. Present data shows that inducible SMO parallel with APAO could playanimportant roleinpolyamine based drug action, i.e. degradation of parent drug and its metabolites, having significant impact on efficiency of these drugs, and hence for the development of novel N-alkylated polyamine analogues.",
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T1 - Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases

AU - Häkkinen, Merja R.

AU - Hyvönen, Mervi T.

AU - Auriola, Seppo

AU - Casero, Robert A

AU - Vepsäläinen, Jouko

AU - Khomutov, Alex R.

AU - Alhonen, Leena

AU - Keinänen, Tuomo A.

PY - 2010/2

Y1 - 2010/2

N2 - N-alkylated polyamine analogues have potential as anticancer and antiparasitic drugs. However, their metabolism in the host has remained incompletely defined thus potentially limiting their utility. Here, we have studied the degradation of three different spermine analogues N,N′-bis-(3-ethylaminopropyl)butane-1,4-diamine (DESPM), N-(3-benzyl-aminopropyl)-N′-(3-ethylamino-propyl)butane-1,4-diamine (BnEtSPM) and N,N′-bis-(3-benzylaminopropyl)butane-1,4-diamine (DBSPM) and related mono-alkylated derivatives as substrates of recombinant human polyamine oxidase (APAO) and spermine oxidase (SMO). APAO and SMO metabolized DESPM to EtSPD [Km(APAO) = 10 μM, kcat(APAO) = 1.1 s -1 and Km(SMO) = 28 μM, kcat(SMO) = 0.8 s-1, respectively], metabolized BnEtSPM to EtSPD [Km(APAO) = 0.9 μM, kcat(APAO) = 1.1 s-1 and Km(SMO) = 51 μM, kcat(SMO) = 0.4 s-1, respectively], and metabolized DBSPM to BnSPD [Km(APAO) = 5.4 μM, k cat(APAO) = 2.0 s-1and Km(SMO) = 33 μM, kcat(SMO) = 0.3 s-1, respectively]. Interestingly, mono-alkylated spermine derivatives were metabolized by APAO and SMO to SPD [EtSPM Km(APAO) =16 μM, kcat(APAO) = 1.5 s -1; Km(SMO) = 25 μM, kcat(SMO) = 8.2 s -1; BnSPM Km(APAO) = 6.0 μM, kcat(APAO) = 2.8 s-1; Km(SMO) =19 μM, kcat(SMO) = 0.8 s-1, respectively]. Surprisingly, EtSPD [Km(APAO) = 37 μM, kcat(APAO) = 0.1 s-1; Km(SMO) =48 μM, kcat(SMO) = 0.05 s-1] and BnSPD [Km(APAO) = 2.5 μM, kcat(APAO) = 3.5 s-1; Km(SMO) =60 μM, kcat(SMO) = 0.54 s-1] were metabolized to SPD by both the oxidases. Furthermore, we studied the degradation of DESPM, BnEtSPM or DBSPM in the DU145 prostate carcinoma cell line. The same major metabolites EtSPD and/or BnSPD were detected both in the culture medium and intracellularly after 48 h of culture. Moreover, EtSPM and BnSPM were detected from cell samples. Present data shows that inducible SMO parallel with APAO could playanimportant roleinpolyamine based drug action, i.e. degradation of parent drug and its metabolites, having significant impact on efficiency of these drugs, and hence for the development of novel N-alkylated polyamine analogues.

AB - N-alkylated polyamine analogues have potential as anticancer and antiparasitic drugs. However, their metabolism in the host has remained incompletely defined thus potentially limiting their utility. Here, we have studied the degradation of three different spermine analogues N,N′-bis-(3-ethylaminopropyl)butane-1,4-diamine (DESPM), N-(3-benzyl-aminopropyl)-N′-(3-ethylamino-propyl)butane-1,4-diamine (BnEtSPM) and N,N′-bis-(3-benzylaminopropyl)butane-1,4-diamine (DBSPM) and related mono-alkylated derivatives as substrates of recombinant human polyamine oxidase (APAO) and spermine oxidase (SMO). APAO and SMO metabolized DESPM to EtSPD [Km(APAO) = 10 μM, kcat(APAO) = 1.1 s -1 and Km(SMO) = 28 μM, kcat(SMO) = 0.8 s-1, respectively], metabolized BnEtSPM to EtSPD [Km(APAO) = 0.9 μM, kcat(APAO) = 1.1 s-1 and Km(SMO) = 51 μM, kcat(SMO) = 0.4 s-1, respectively], and metabolized DBSPM to BnSPD [Km(APAO) = 5.4 μM, k cat(APAO) = 2.0 s-1and Km(SMO) = 33 μM, kcat(SMO) = 0.3 s-1, respectively]. Interestingly, mono-alkylated spermine derivatives were metabolized by APAO and SMO to SPD [EtSPM Km(APAO) =16 μM, kcat(APAO) = 1.5 s -1; Km(SMO) = 25 μM, kcat(SMO) = 8.2 s -1; BnSPM Km(APAO) = 6.0 μM, kcat(APAO) = 2.8 s-1; Km(SMO) =19 μM, kcat(SMO) = 0.8 s-1, respectively]. Surprisingly, EtSPD [Km(APAO) = 37 μM, kcat(APAO) = 0.1 s-1; Km(SMO) =48 μM, kcat(SMO) = 0.05 s-1] and BnSPD [Km(APAO) = 2.5 μM, kcat(APAO) = 3.5 s-1; Km(SMO) =60 μM, kcat(SMO) = 0.54 s-1] were metabolized to SPD by both the oxidases. Furthermore, we studied the degradation of DESPM, BnEtSPM or DBSPM in the DU145 prostate carcinoma cell line. The same major metabolites EtSPD and/or BnSPD were detected both in the culture medium and intracellularly after 48 h of culture. Moreover, EtSPM and BnSPM were detected from cell samples. Present data shows that inducible SMO parallel with APAO could playanimportant roleinpolyamine based drug action, i.e. degradation of parent drug and its metabolites, having significant impact on efficiency of these drugs, and hence for the development of novel N-alkylated polyamine analogues.

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KW - Polyamine oxidase

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KW - Spermine oxidase

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