New oxaanalogues of spermine

Article abstract of DOI:10.1007/s11171-005-0026-4

A new isosteric charge-deficient spermine analogue, 1,12-diamino-4,9-diaza- 5-oxadodecan, and O-(7-amino-4-azaheptyl)oxime of 3-aminopropanal, a stable analogue of the Schiff base intermediate in the enzymatic oxidation of spermine, were synthesized. The possible use of these compounds for the inhibition of spermine oxidase is discussed.

Full text of DOI:10.1007/s11171-005-0026-4

Russian Journal of Bioorganic Chemistry, Vol. 31, No. 2, 2005, pp. 189–195. Translated from Bioorganicheskaya Khimiya, Vol. 31, No. 2, 2005, pp. 206–212.
Original Russian Text Copyright © 2005 by Khomutov, Simonyan, Vepsalainen, Keinanen, Alhonen, Janne.
New Oxaanalogues of Spermine
1
A. R. Khomutov*, A. R. Simonyan*, J. Vepsalainen**, T. A. Keinanen***,
L. Alhonen***, and J. Janne***
* Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, ul Vavilova 32, Moscow, 119991 Russia
** Department of Chemistry, University of Kuopio, P.O. Box 1627, Kuopio, FIN-70211 Finland
*** Virtanen Institute for Molecular Sciences, University of Kuopio, P.O. Box 1627, Kuopio, FIN-70211 Finland
Received May 25, 2004; in final form, September 28, 2004
Abstract—A new isosteric charge-deficient spermine analogue, 1,12-diamino-4,9-diaza-5-oxadodecan, and
O-(7-amino-4-azaheptyl)oxime of 3-aminopropanal, a stable analogue of the Schiff base intermediate in the
enzymatic oxidation of spermine, were synthesized. The possible use of these compounds for the inhibition of
spermine oxidase is discussed.
Key words: oxaspermine, polyamine analogues, polyamines, spermine oxidase
1
INTRODUCTION
Therefore, the Spm degradation can proceed by two
independent pathways (Scheme 1). Several SMO inhib-
itors are known; the best of them, cis-
3,8,13,18,23,28,33,38,43,48-decaazapentacontene-25
(SL-11150, figure), has IC50 10–7 M [8]. A similar fla-
vin-dependent Spm oxidase also exists in plants,
whereas 1,17-bis(guanidyl)-9-azaheptadecane (guaza-
tine, figure) inhibits the enzyme from maize with Ki
A selective control of activity of the enzymes
involved in the metabolism of polyamines is widely
used at studying the role of spermine and spermidine in
2
the processes of growth and differentiation of cells.
These investigations are of a certain scientific and
applied importance due to significance and a variety of
cellular functions of polyamines and their increased
content in tumor cells in comparison with that on nor-
mal cells [1, 2].
7.5 × 10–9 M [12]. The X-ray study of the enzyme–
inhibitor complex (E–I) showed that the inhibition effi-
ciency is due to a firm noncovalent binding of this com-
pound in the active site of enzyme [13].
The activation of the enzymes of polyamine catabo-
lism is the most effective way of the exhaustion of the
intracellular pool of spermine and spermidine [3].
Therefore, in recent years, the study of these enzymes
is one of important directions in the polyamine bio-
chemistry.
The classical pathway of spermine degradation con-
sists in its N1-acetylation catalyzed by SSAT and the
subsequent oxidation of the product by PAO (Scheme
1) [4]. A selective effectively acting nonreversible
inhibitor N1,N4-bis(2,3-butadienyl)-1,4-diaminobutane
(MDL-72.527, see the figure) is known for the well-
known flavin-dependent PAO; it exhibits Ki 0.09 µM
and τ1/2 2.2 min toward to the isolated enzyme [5].
The syntheses of two new compounds, 1,12-
diamino-4,9-diaza-5-oxadodecane (I) (Scheme 2) and
O-(7-amino-4-azaheptyl)oxime of 3-aminopropanal
(II) (Scheme 3), are described in this communication;
we suggest the use of these compounds for the SMO
inhibition and the study of the mechanism of action of
this enzyme.
RESULTS AND DISCUSSION
The enzymatic oxidation of Spm was shown to pro-
ceed through the intermediate formation of a Schiff
base, hydrolyzed to 3-aminopropanal and Spd (Scheme
4) [10].
Recently, SMO has been found in animal cells; this
is a new flavin-dependent enzyme that oxidizes Spm
into Spd without the preliminary N1-acetylation [6–11].
Here we describe the substitution of oxygen atom
for the methylene group of Spm in position 5, which
resulted in N,O-dialkylhydroxylamine (I), in order to
obtain new potential inhibitors of SMO. Since oxa-Spm
(I) is an isostere of Spm, one can expect that this com-
pound will be a good competitive inhibitor of SMO.
Oxygen atom in the fifth position of (I) decreases the
basicity of the adjacent NH group (pKa ~ 5.5) in com-
parison with Spm (pKa of the NH group of 7.97 [14]),
and, therefore, (I) exists in the form of trication at the
1
Corresponding author; phone: +7 (095) 135-6065; fax: +7 (095)
135-1405; e-mail: alexkhom@genome.eimb.relarn.ru
Abbreviations: Bpoc, 2-(4-biphenyl)prop-2-yloxycarbonyl; Mts,
2
mesitylenesulfonyl; Ns, 2-nitrobenzenesulfonyl; PAO, polyamine
oxidase (EC 1.5.3.11); Pmc, 2,2,5,7,8-pentamethyl-6-chromane-
sulfonyl; Spd, spermidine (1,8-diamino-4-azaoctane); Spm, sper-
mine (1,12-diamino-4,9-diazadodecane); SMO, spermine oxi-
1
dase; and SSAT, spermine/spermidine N -acetyltransferase (EC
2.3.1.57).
1068-1620/05/3102-0189 © 2005 Pleiades Publishing, Inc.

190
KHOMUTOV et al.
H
NH
NH
NH
NH
4
·
N
N
H
·
4
MDL-72.527
SL-11150
NH
NH
NH
NH
NH
H2N
NH2
Guazatine
The SMO and PAO inhibitors
physiological pH value. Deprotonation of the second- new information on the mechanism of the SMO action
and the methods of regulation of its activity.
ary amino group of Spm that proceeds in the active site
of SMO precedes its oxidation, and oxa-Spm (I) could
The general method of the synthesis of O,N-disub-
hence be considered as a peculiar analogue of the tri- stituted hydroxylamines is the reduction of O-substi-
tuted oximes with complex borohydrides in a moder-
ately acidic medium (pH ~3). The 1,8-diamino-4-aza-
5-oxaoctane analogue of Spd was obtained by this
method starting from 3-azidopropanal and 3-ami-
nooxy-1-(N-benzyloxycarbonyl)aminopropane [16].
An alternative scheme of synthesis of 1,8-diamino-
4-aza-5-oxaoctane consists in the condensation of Pmc
derivative of N-(3-aminooxypropyl)phthalimide with 3-
N-Bpoc-aminopropanol under the conditions of Mit-
sunobu reaction, followed by the removal of protective
groups [17]. The preparation of the bisoxaanalogue of
Spm, 1,12-diamino-4,9-diaza-5,8-dioxadodecane, is
based on the interaction of 3-N-Bpoc-aminopropanol
with bis-Pmc-derivative of 1,2-diaminooxyethane [17].
charged form of Spm intermediately arising in the
active site of SMO. The formation of covalent adduct
with coenzyme, similar to that described for the oxida-
tion of 1-cycloheptylmethyl-12-ethylnorspermine
(CHENSpm) by the enzyme from maize [13], cannot
be excluded at the enzymatic oxidation of (I). If (I)
would be oxidized like Spm, oxime (II), a stable ana-
logue of the Schiff base, should be formed in the active
site of SMO in situ (Scheme 4). The obtaining of stable
analogues of intermediates in enzymatic reactions is
known to be the rather universal algorithm of the design
of effective inhibitors of various enzymes (cf. the
review [15]). Therefore, in this work, we describe also
the preparation of oxime (II) along with the synthesis
of (I). A comparative study of the interaction of these
The pathway whose key stage was the alkylation of
2-nitrobenzenesulfonyl derivative of 1,4-bis(benzyl-
compounds with SMO would allow the obtaining of a oxycarbonyl)-7-aminooxy-4-aza-1-aminoheptane (V)
NH2
H2N
3
3
H
N
Put
NHAc
H2N
N1-Ac-Spd
H
N
NH2
H2N
H
N
Spd
1
NH2
AcHN
N
H
H
N
N1-Ac-Spm
NH2
H2N
N
H
Spm
1
Scheme 1. Catabolism of polyamines [4]. 1, SMO (spermine oxidase); 2, SSAT (spermine/spermidine N -acetyltransferase); 3, PAO
(polyamine oxidase); and Put, putrescine.
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 31 No. 2 2005

NEW OXAANALOGUES OF SPERMINE
Cbz
191
Cbz
N
Me
i
O
N
NHCbz
Et
O
NHCbz
H2N
O
N
(III)
(IV)
Cbz
Cbz
N
ii
iii
O
N
NHCbz
O
NHCbz
NsHN
BocNH
N
(V)
(VI)
Ns
Cbz
N
H
N
v
iv
O
NHCbz
O
NH2 · 4HB
BocNH
N
H2N
N
H
H
(VII)
(I)
i, HCl/i-PrOH/H2O, ii, Ns-Cl/CH2Cl2, iii, BocNH(CH2)3I/K2CO3/DMF, iv, PhSH/K2CO3/DMF, v, HBr/AcOH.
Scheme 2. The scheme of synthesis of 1,12-diamino-4,9-diaza-5-oxadodecane (I).
i
ii
BocNH
NH2
OH
(VIII)
NH2
OH
BocNH
O
iv
BocNH
N
NH2
N
O
iii
O
N
N
H
NH
2 · 3HCl
H
(IX)
(II)
i, Boc2O/THF, ii, CrO3 · Py · HCl/Al2O3/CH2Cl2, iii, H2NO(CH2)3NH(CH2)3NH2/MeOH/H2O, iv, HCl/MeOH.
Scheme 3. The scheme of synthesis of O-(7-amino-4-azaheptyl)oxime of 3-aminopropanal (II).
with 3-N-Boc-aminopropyl iodide was chosen for the tion of (V) with 3-N-Boc-aminopropyl iodide, selective
denosylation of (VI) with thiophenol (like that
described for amines [19, 20], and the removal of Cbz
groups by HBr/AcOH, which resulted in the target tet-
rahydrobromide of 1,12-diamino-4,9-diaza-5-oxa-
dodecane (I).
The synthesis of oxime (II) was achieved starting
from 3-N-Boc-aminopropanol (VIII) (Scheme 3),
which was oxidized at the first stage by pyridinium
chlorochromate on neutral alumina to the correspond-
ing aldehyde (Scheme 3). 3-(N-tert-Butyloxycarbo-
nyl)aminopropanal was not isolated in pure state, but
was directly subjected to the reaction with 1-amino-4-
aza-7-aminooxyheptane. The resulting oxime (IX) was
isolated by column chromatography on silica gel as a
obtaining of 1,12-diamino-4,9-diaza-5-oxadodecane
(I) (Scheme 2).
The gem-bis-Mts-derivative of O-substituted
hydroxylamine is known to be formed under the stan-
dard conditions of the synthesis of arenesulfonylamides
with the use of triethylamine as a base and only at an
excess mesitylenesulfonyl chloride (Mst-Cl) [18]. The
interaction of 2-nitrobenzenesulfonyl chloride (Ns-Cl)
with hydroxylamine (IV) proceeds ambiguously even
at the equimolar ratio Ns-Cl : RONH2 : Et3N. However,
the reaction with 2.5–3.0-fold excess of O-substituted
hydroxylamine in the absence of other bases leads to
mono-NS-derivative (V) in yield of approximately
70%. The further conversions consisted in the alkyla- mixture of syn- and anti-isomers at the ratio of 40 : 60
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 31 No. 2 2005

192
KHOMUTOV et al.
H
N
(a)
H2N
H2N
O
+
NH2
Spd
H
N
H
N
SMO
H2N
H2N
N
H
NH2
N
NH2
FAD
O2
FADH–
H2O2
Spm
(b)
H
N
H
N
SMO
H2N
O
H2N
O
N
H
NH2
N
NH2
FAD
O2
FADH–
H2O2
(I)
(II)
H
N
H2N
O
H2N
O
+
NH2
Scheme 4. (a) Oxidative cleavage of spermine in animals [10], and (b) the presumed enzymatic oxidation of (I) to oxime (II).
(1H NMR data). The subsequent removal of Boc-pro-
tective group by the treatment with HCl/MeOH at 0°C
and recrystallization led to the desired (II) trihydro-
chloride; in this case, the ratio of syn- and anti-isomers
remained almost unchanged (35 : 65).
umn chromatography was carried out on Kieselgel (40–
63 µm, Merck), elution systems being specified in text.
NMR spectra were registered on a Bruker Avance
500 DRX instrument (Germany) with the working fre-
1
quency of 500.1 MHz for H and 125.8 MHz for 13C
Note that the reduction of (II) with NaCNBH3
should lead to oxa-Spm (I), because the reduction of
oximes is the general method of synthesis of O,N-
dialkylhydroxylamine (see above).
nuclei. Tetramethylsilane was used as an internal stan-
dard in CDCl3 and CD3OD solutions and sodium salt of
3-trimethylsilyl-1-propanesulfonic acid in D2O. Chem-
ical shifts are given in ppm, and spin–spin coupling
constants in Hz.
EXPERIMENTAL
7-Aminooxy-4-(N-benzyloxycarbonyl)aza-1-(N-
benzyloxycarbonyl)aminoheptane (IV). Concentrate
HCl (1.5 ml) was added to a solution of (III) (4.7 g,
9.73 mmol) in isopropanol (10 ml), the solution was
kept for 10 min at room temperature and evaporated in
a vacuum to dryness. The residue was dissolved in
water (5 ml), 10 M NaOH (1.5 ml) was added, and
extracted with chloroform (3 × 5 ml). The extract was
washed with a saturated solution of NaCl (2 × 2 ml),
dried with MgSO4, and evaporated in a vacuum to dry-
ness. The residue was dissolved in chloroform (6 ml)
and purified by column chromatography on silica gel
(60 g), successively eluting with chloroform and 97 : 3
chloroform–methanol. The target (IV) was dried in a
vacuum over ê2é5; viscous oil; yield 2.71 g (73%); Rf
The reagents Cbz-Cl, Boc2O, PhSH, absolute EtOH,
and 1-amino-3-bromopropane were from Fluka; 2-
nitrobenzenesulfonyl chloride and 3-aminopropanol,
from Aldrich. 7-Aminooxy-4-aza-1-aminoheptane and
7-(1'-ethoxyethylidene)aminooxy-4-(N-benzyloxycar-
bonyl)aza-1-(N-benzyloxycarbonyl)aminoheptane were
synthesized as described in [21], and pyridinium chlo-
rochromate adsorbed on Al2O3 was synthesized as
described in [22].
TLC was carried out on precoated Kieselgel 60 F254
plates (Merck) in the following systems: (A) 95 : 5
chloroform–methanol, (B) chloroform, (C) 4 : 2 : 1 : 2
n-butanol–acetic acid–pyridine–water, (D) 9 : 1 chloro-
form–methanol, and (E) 8 : 2 dioxane–25% ammonia.
Substances were detected on the chromatograms by
their absorption of UV light and by the color reaction
with ninhydrin; the aminooxy compounds, in the form
of fluorescent oximes of pyridoxal-5'-phosphate. Col-
1
0.37 (A); H NMR (CDCl3): 7.4–7.2 (10 H, m, Ph);
5.12 (2 H, s, CH2Ph), 5.08 (2 H, s, CH2Ph), 3.77 (2 H,
m, H2NOCH2), 3.46 (2 H, m, H2NOCH2CH2CH2N),
3.35–3.08 [4 H, m, NCH2(CH2)2NHCbz
+
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 31 No. 2 2005

NEW OXAANALOGUES OF SPERMINE
193
N(CH2)2CH2NHCbz], and 1.65–1.38 (4 H, m, in chloroform, successively washed with water (2 ×
5 ml) and a saturated solution of NaCl (2 × 5 ml), and
dried with MgSO4. The solution was evaporated in a
vacuum to dryness, and the raw 1-(N-tert-butyloxycar-
bonyl)amino-4-(N-2-nitrobenzenesulfonyl)aza-5-oxa-
9-(N-benzyloxycarbonyl)aza-12-(N-benzyloxycarbo-
nyl)aminododecane (VI) was dissolved in DMF
(30 ml). The solution was mixed with K2CO3 (1.2 g,
8.72 mmol) and PhSH (0.27 ml, 2.6 mmol) and stirred
for 4 h at room temperature. The reaction mixture was
evaporated to dryness in a vacuum, the residue was sus-
pended in chloroform, and the precipitate was sepa-
rated by centrifugation and washed with a small quan-
tity of chloroform. The combined organic solutions
were successively washed with water (5 ml), 10% citric
acid (2 × 10 ml), 1 M NaHCO3 (2 × 10 ml), and water
(10 ml) and evaporated in a vacuum. The residue was
dissolved in 100 : 1 chloroform–methanol mixture
(6 ml) and purified by a chromatography on a silica gel
column (60 g), eluted with 100 : 1 chloroform–metha-
nol mixture to get (VII) as a viscous oil dried in a vac-
H2NOCH2CH2CH2N + NCH2CH2CH2NHCbz).
7-(N-2-Nitrobenzenesulfonyl)aminooxy-4-(N-ben-
zyloxycarbonyl)aza-1-(N-benzyloxycarbonyl)amino-
heptane (V). A solution of Ns-Cl (0.48 g, 2.2 mmol) in
dichloromethane (6.5 ml) was added to a solution of
(IV) (2.71 g, 6.5 mmol) in dichloromethane (8 ml). The
reaction mixture was kept for 24 h at room temperature
and evaporated in a vacuum to dryness. The residue was
dissolved in 99 : 1 chloroform–methanol mixture (6 ml)
and purified on a silica gel column (60 g) eluted with
99 : 1 chloroform–methanol. The target (V) was dried
in a vacuum over ê2é5 to give a viscous oil; yield 1.3 g
(65% from Ns-Cl); Rf 0.57 (A); 1H NMR (CDCl3): 8.18
(1 H, m, Ns), 7.99 (1 H, m, Ns), 7.85 (1 H, m, Ns), 7.74
(1 H, m, Ns), 7.36–7.26 (10 H, m, Ph), 5.60 (1 H, s,
HNO), 5.12 (2 H, s, CH2Ph), 5.08 (2 H, s, CH2Ph), 4.04
(2 H, m, NsHNOCH2), 3.30–3.28 (4 H, m,
H2NOCH2CH2CH2N + NCH2(CH2)2NHCbz), 3.13 (2
H, m, N(CH2)2CH2NHCbz), and 1.88–1.67 (4 H, m,
NsHNOCH2CH2CH2N + NCH2CH2CH2NHCbz).
1
uum over ê2é5; yield 0.58 g (46%); Rf 0.37 (A); H
1-(N-tert-Butyloxycarbonyl)amino-4-aza-5-oxa-9-
(N-benzyloxycarbonyl)aza-12-(N-benzyloxycarbo-
nyl)aminododecane (VII). A cooled to +4°C solution
of 3-bromo-1-aminopropane hydrobromide (5.5 g,
25 mmol) in water (7 ml) was mixed with 10 M NaOH
(2.5 ml), dioxane (20 ml) and a solution of Boc2O (6 g,
28 mmol) in dioxane (10 ml). The reaction mixture was
stirred for 16 h at room temperature, evaporated to dry-
ness in a vacuum, and the residue was dissolved in chlo-
roform. The solution was successively washed with
water (2 × 10 ml), 10% citric acid (2 × 10 ml), and water
(3 × 10 ml), dried with MgSO4, and evaporated to dry-
ness in a vacuum. The residue was dissolved in acetone
(20 ml), NaI (4.1 g, 27.2 mmol) was added, and the
mixture was stirred for 18 h at room temperature. The
NMR (CDCl3): 7.36–7.26 (10 H, m, Ph), 5.12 (2 H, s,
CH2Ph), 5.09 (2 H, s, CH2Ph), 3.62 (2 H, m,
HNOCH2(CH2)2NCbz), 3.34–3.28 CH2N(Cbz)CH2);
3.23–3.11 (4 H, m, BocNHCH2 + CH2NHCbz), 2.86
BocNH(CH2)2CH2NHO); 1.78–1.60 (6 H, m,
BocNHCH2CH2CH2NHO + HNOCH2CH2CH2N +
NCH2CH2CH2NHCbz), and 1.41 [9 H, s, C(CH3)3).
1,12-Diamino-4,9-diaza-5-oxadodecane tetrahy-
drobromide (I). A 32.3% solution of HBr in AcOH
(3 ml) was added to a solution of (VII) (0.58 g,
1.01 mmol) in AcOH (5 ml). The mixture was kept at
room temperature until the evolution of CO2 ceased.
The precipitated solid was separated, washed with
diethyl ether, and dried in a vacuum over P2O5/KOH to
precipitate was separated, the filtrate was evaporated to give (I); yield 0.36 g (69%); Rf 0.27 (B); mp 169–171°C
1
dryness in a vacuum, and the residue was dissolved in
chloroform. The solution was washed with water (2 ×
2 ml), dried with MgSO4, and evaporated, and the resi-
due was dried in a vacuum at +4°C over P2O5 to give
1-(N-tert-butyloxycarbonyl)amino-3-iodopropane in
the form of a viscous slowly solidifying yellowish oil;
yield 5.35 g (83%). A part of the semicrystalline prod-
uct was triturated with a small quantity of hexane and
kept overnight at +4°C; the solid was separated, washed
with cold hexane, dried in a vacuum over paraffin and
P2O5, and obtained colorless crystals of BocNH(CH2)3I;
(decomp.); H NMR (D2O): 4.11 (2 H, t, J 6.0,
HNOCH2), 3.31 (2 H, t, J 7.6, OHNCH2CH2CH2NH2),
3.17 J 7.9, HNO(CH2)2CH2NH2); 3.14 J 8.0,
CH2HN(CH2)2CH2NH2); 3.08 (2 H, t,
HNOCH2CH2CH2), 3.07 [2 H, t,
J
J
7.4,
7.4,
CH2HNCH2(CH2)2NH2); and 2.10–2.01 (6 H, m,
OHNCH2CH2CH2NH2
+
HNOCH2CH2
+
CH2HNCH2CH2CH2NH2); 13C NMR (D2O): 73.83 (t,
HNOCH2), 49.84 (t, OHNCH2CH2CH2NH2), 47.79 (t,
HNOCH2CH2CH2), 47.62 [t, CH2HNCH2(CH2)2NH2],
1
40.09
[t,
HNO(CH2)2CH2NH2],
39.52
[t,
Rf 0.36 (B); mp 41–42°C; H NMR (CD3OD): 4.58
CH2HN(CH2)2CH2NH2], 27.48 (t, HNOCH2CH2),
26.73 (t, CH2HNCH2CH2CH2NH2), and 25.56 (t,
OHNCH2CH2CH2NH2).
(1 H, br. s, NH), 3.26 (2 H, m, NHCH2), 3.16 (2 H, t, J
6.5, CH2I), 2.06–1.98 (2 H, m, CH2CH2CH2), and 1.48
[9 H, s, C(CH3)3].
Found, %: C 20.51, H 5.28, N 10.50. C9H28Br4N4O.
Calc., %: C 20.47, H 5.35, N 10.61.
3-(N-tert-Butyloxycarbonyl)aminopropanol (VIII).
A solution of Boc2O (1.5 g, 7 mmol) in THF (7 ml) was
A mixture of (V) (1.3 g, 2.2 mmol), K2CO3 (1.19 g,
8.64 mmol), and BocNH(CH2)3I (0.8 g, 2.8 mmol) was
stirred in DMF (10 ml) for 24 h at room temperature;
the residue was separated; and the filtrate was evapo-
rated in a vacuum to dryness. The residue was dissolved added to a solution of 3-aminopropanol (0.5 g, 7 mmol)
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 31 No. 2 2005

194
KHOMUTOV et al.
1
in THF (7 ml). The mixture was kept for 12 h at room
temperature and evaporated in a vacuum to dryness.
The residue was dissolved in chloroform; the solution
was successively washed with 10% citric acid (2 × 5
ml), 1 M NaHCO3 (2 × 5 ml), and water (5 ml); and was
evaporated in a vacuum to dryness. The residue was
dried in a vacuum over ê2é5, to get (VIII) as a viscous
mp 193−194°C (decomp.); isomer I: H NMR (D2O):
7.59 (1 H, t, J 5.3, CH=NO), 4.18 (2 H, t, J 6.0,
NOCH2), 3.26 (2 H, t, J 7.0, NH2CH2CH2CH=NO),
3.22–3.16 [4 H, m, NHCH2(CH2)2NH2
NO(CH2)2CH2NH], 3.12 (2 H, t,
NHCH2CH2CH2NH2), 2.65 (2 H, dt,
NH2CH2CH2CH=NO), and 2.15–2.04 (4 H, m,
NHCH2CH2CH2NH2 + NOCH2CH2CH2NH); 13C NMR
(D2O): 152.58 (d, N=CH), 73.24 (t, CH2O), 48.10 [t,
CH2(CH2)2O], 47.48 [t, NH2(CH2)2CH2], 39.43 [t,
NH2CH2(CH2)2], 39.10 (t, NH2CH2CH2CH=N), 29.86
(t, CH2CH=N), 28.04 (t, CH2CH2O), and 26.57 (t,
NH2CH2CH2CH2); isomer II: H NMR (D2O): 6.96
(1 H, t, J 5.5, CH=NO), 4.23 (2 H, t, J 6.0, NOCH2),
3.24 (2 H, t, J 7.3, NH2CH2CH2CH=NO), and 2.77 (2
H, dt, J 7.3, NH2CH2CH2CH=NO); 13C NMR (D2O):
151.95 (d, N=CH), 73.34 (t, CH2O)), 47.96
[t, CH2(CH2)2O], 47.45 [t, NH2(CH2)2CH2]; 39.14
(o, NH2CH2CH2CH=N); 28.26 (t, CH2CH2O), 26.68
(t, CH2CH=N), and 26.60 (t, NH2CH2CH2CH2); chemi-
cal shifts of other nuclei coincide with those of isomer
I. The isomer I/isomer II ratio was 65 : 35.
+
5.7,
7.0,
J
J
oil; yield 1 g (82%); Rf 0.5 (D); 1H NMR (CDCl3): 4.82
(1 H, s, NH), 3.65 (2 H, m, CH2OH), 3.27 (2 H, m,
NHCH2), 3.08 (1 H, s, OH), 1.65 (2 H, m, CH2CH2OH),
and 1.43 [9 H, s, C(CH3)3].
O-(7-Amino-4-aza)heptyloxime of 3-(N-tert-
butyloxycarbonyl)aminopropanal (IX). Pyridinium
chlorochromate on neutral Al2O3 (7 g) was added to a
solution of (VIII) (0.7 g, 4 mmol) in anhydrous ben-
zene (40 ml). The mixture was stirred for 4 h at 20°C,
the precipitate was separated, and the filtrate was evap-
orated in a vacuum to dryness. The residue was dis-
solved in methanol (5 ml) and treated with a neutral
solution of 7-aminooxy-4-aza-1-aminoheptane trihy-
drochloride (0.256 g, 1 mmol) in 50% methanol
(10 ml). The reaction mixture was stirred for 2 h at
room temperature, evaporated to dryness in a vacuum,
and the residue was dissolved in 95 : 5 dioxane–25%
ammonia (6 ml). The purification on a silica gel column
(60 g) successively eluted with 95 : 5 dioxane–25%
ammonia and 8 : 2 dioxane–25% ammonia gave, after
drying the target fraction in a vacuum over ê2é5, 0.2 g
(66% from 7-aminooxy-4-aza-1-aminoheptane trihy-
1
Found, %: C 34.84, H 8.11, N 17.54. C9H25Cl3N4O.
Calc., %: C 34.68, H 8.08, N 17.98.
ACKNOWLEDGMENTS
This work was supported by the Russian Foundation
of basic Research (project no. 03-04-49080) and by the
Academy of Sciences of Finland.
1
drochloride) of (IX) as a viscous oil; Rf 0.5 (E). H
NMR of isomer I (CDCl3): 7.38 (1 H, t, J 5.4,
NHCH2CH2CH=NO), 4.09 (2 H, t,
NOCH2CH2CH2NH), 3.32 (2 H,
NHCH2CH2CH=NO), 2.79 (2 H, t,
J
br.
J
6.2,
m,
6.3,
REFERENCES
NHCH2CH2CH2NH2), 2.70 [4 H, m, NHCH2(CH2)2NH2
1. Seiler, N., Curr. Drug Targets, 2003, vol. 4, pp. 537–564.
2. Seiler, N., Curr. Drug Targets, 2003, vol. 4, pp. 565–585.
+ NO(CH2)2CH2NH], 2.37 (2 H, dt,
J
5.9,
m,
NHCH2CH2CH=NO),
NOCH2CH2CH2NH),
1.83
1.66
(2
(2
H,
H,
3. Casero, R.A. and Woster, P.M., J. Med. Chem., 2001,
m,
vol. 44, pp. 1–26.
NHCH2CH2CH2NH2), and 1.44 [9 H, s, C(CH3)3]; iso-
mer II: 1H NMR (CDCl3): 6.69 (1 H, t, J 5.5,
4. Cohen, S.S., A Guide to the Polyamines, New York:
Oxford Univ. Press, 1998.
NHCH2CH2CH=NO), 4.14 (2 H, t,
NOCH2CH2CH2NH), 3.28 (2 H,
NHCH2CH2CH=NO), 2.51 (2 H, dt,
NHCH2CH2CH=NO), and 1.86 (2 H, m,
NOCH2CH2CH2NH); chemical shifts of other protons
coincide with those of isomer I. The isomer I/isomer II
ratio was 60 : 40.
J
br.
J
6.1,
m,
5.9,
5. Bey, P., Bolkenius, F.N., Seiler, N., and Casara, P.,
J. Med. Chem., 1985, vol. 28, pp. 1–2.
6. Wang, Y., Devereux, W., Woster, P.M., Murray-Stew-
art, T., Hacker, A., and Casero, R.A., Cancer Res., 2001,
vol. 61, pp. 5370–5373.
7. Murray-Stewart, T., Wang, Y., Devereux, W., and
Casero, R.A., Biochem. J., 2002, vol. 368, pp. 673–677.
8. Wang,Y., Murray-Stewart, T., Devereux, W., Hacker, A.,
Frydman, B., Woster, P.M., and Casero, R.A., Biochem.
Biophys. Res. Commun., 2003, vol. 304, pp. 605–611.
9. Devereux, W., Wang,Y., Murray-Stewart, T., Hacker, A.,
Smith, R., Frydman, B., Valasinas, A.L., Reddy, V.K.,
Marton, L.J., Ward, T.D., Woster, P.M., and Casero, R.A.,
Cancer Chem. Pharm., 2003, vol. 52, pp. 383–390.
10. Vujcic, S., Diegelman, P., Bacchi, C.J., Kramer, D.L.,
and Porter, C.W., Biochem. J., 2002, vol. 367, pp. 665–
675.
O-(7-Amino-4-azaheptyl)oxime of 3-aminopro-
panal trihydrochloride (II). A solution of (IX)
(0.15 g, 0.5 mmol) in absolute methanol (5 ml) cooled
to 0°C was treated with 4 M HCl/MeOH (5 ml) and
kept overnight at 4°C. The reaction mixture was evapo-
rated to dryness in a vacuum, and the residue was
coevaporated with MeOH, recrystallized from MeOH–
EtOH, and dried in a vacuum over P2O5/KOH. Crystal-
line (II) was obtained; yield 0.106 g (68%); Rf 0.1 (E);
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 31 No. 2 2005

NEW OXAANALOGUES OF SPERMINE
195
11. Cervelli, M., Polticelli, F., Federico, R., and Mariot- 19. Siaugue, J.-M., Segat-Dioury, F., Favre-Reguillon, A.,
tini, P., J. Biol. Chem., 2003, vol. 278, pp. 5271–5276.
Madic, Ch., Foos, J., and Guy, A., Tetrahedron Lett.,
2000, vol. 41, pp. 7443–7446.
12. Federico, R., Leone, L., Botta, M., Binda, C., Ange-
lini, R., Venturini, G., and Ascenzi, P., J. Enzyme Inhib.,
2001, vol. 16, pp. 147–155.
20. Fukuyama, T., Jow, Ch.K., and Cheung, M., Tetrahedron
Lett., 1995, vol. 36, pp. 6373–6374.
13. Binda, C., Angelini, R., Federico, R., Ascenzi, P., and
Mattevi, A., Biochemistry, 2001, vol. 40, pp. 2766–2776.
21. Khomutov, A.R., Vepsalainen, J., Shvetsov, A.S.,
Hyvonen, T., Keinanen, T.A., Pustobaev, V.N., Elo-
ranta, T.O., and Khomutov, R.M., Tetrahedron, 1996,
vol. 52, pp. 13 751–13 766.
14. Templeton, D.M. and Sarkar, B., Can. J. Chem., 1985,
vol. 63, pp. 3122–3128.
15. Wolfenden, R., Annu. Rev. Biophys. Bioeng., 1976,
vol. 5, pp. 271–306.
22. Tietze, L.F. and Eicher, T., Reactionen und Synthesen im
Organisch-chemischen Praktikum und Forschungslabo-
ratorium, Suttgart: Georg Thieme, 1991. Translated
under the title Preparativnaya organicheskaya khimiya.
Reaktsii i sintezy v praktikume organicheskoi khimii i
nauchno-issledovatel’skoi laboratorii, Moscow: Mir,
1999.
16. Lee, Y.B., Park, M.H., and Folk, J.E., J. Med. Chem.,
1995, vol. 38, pp. 3053–3061.
17. Lin, P.K., Maguire, N.M., and Brown, D.M., Tetrahe-
dron Lett., 1994, vol. 35, pp. 3605–3608.
18. Kuksa, V., Buchan, R., and Lin, P.K., Synthesis, 2000,
no. 9, pp. 1189–1207.
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 31 No. 2 2005