New syntheses of α-methyl- and α,α′- dimethylspermine

Article abstract of DOI:10.1007/s11171-005-0025-5

α-Methylspermine and α,α′-dimethylspermine were synthesized in high overall yields starting from N-(benzyloxycarbonyl)-3- aminobutanol in order to study polyamine biochemistry in vitro and in vivo.

Full text of DOI:10.1007/s11171-005-0025-5

Russian Journal of Bioorganic Chemistry, Vol. 31, No. 2, 2005, pp. 183–188. Translated from Bioorganicheskaya Khimiya, Vol. 31, No. 2, 2005, pp. 200–205.
Original Russian Text Copyright © 2005 by Grigorenko, Vepsalainen, Jarvinen, Keinanen, Alhonen, Janne, Khomutov.
New Syntheses of a-Methyl- and a,a'-Dimethylspermine
N. A. Grigorenko*, J. Vepsalainen**, A. Jarvinen***, T. A. Keinanen***, L. Alhonen***,
1
J. Janne***, and A. R. Khomutov*
* 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 April 8, 2004; in final form, May 14, 2004
Abstract—α-Methylspermine and α,α'-dimethylspermine were synthesized in high overall yields starting
from N-(benzyloxycarbonyl)-3-aminobutanol in order to study polyamine biochemistry in vitro and in vivo.
Key words: α,α'-dimethylspermine, α-methylspermine, polyamines, spermine/spermidine N1-acetyltransferase
INTRODUCTION
homologues that are symmetrically (e.g., DENSpm) or
asymmetrically bisalkylated at terminal amino groups.
Significant amounts of spermidine and spermine are
present in all cell types and are essential for their nor-
These compounds are actively transported into cells
like Spm and Spd; however, they cannot fulfill the vital
cellular functions of polyamines. The accumulation of
DENSpm or similar compounds results in the induction
of SSAT biosynthesis, which catalyses the key step of
polyamine catabolism. The subsequent oxidation of N1-
Ac-Spm to Spd and N1-Ac-Spd to Put results in the
depletion of Spm/Spd pool and inhibition of cell
2
mal growth. Alkyl derivatives of Spd and Spm are
widely used to control the enzymes of polyamine
metabolism and to study the functions of Spm and Spd.
Two groups of the analogues differing in the position of
alkyl substituents are known. The first and the most
studied group includes derivatives of Spm and its growth (see reviews [1, 2]).
NH
NH2
NH
NH
NH
NH
NH
H2N
Spm
DENSpm
The analogues with alkyl substituents at carbon these compounds can induce the biosynthesis of SSAT
[6]. α-MeSpd, α-MeSpm, and α,α'-Me2Spm interact
atoms of polyamine backbone constitute the second
group of polyamine derivatives [3–5]; this is less inves-
with DNA as the natural polyamines [7]. In addition,
tigated. In contrast to N-alkyl derivatives of Spm and α-MeSpd is the substrate of deoxyhypusine synthase,
which executes a posttranslational modification of ini-
tiation factor eIF-5A necessary for the normal function-
ing of ribosome [8]. We have recently reported that α-
MeSpd can prevent the development of acute pancreati-
tis caused by the depletion of polyamine pool resulting
from the SSAT overinduction in SSAT-transgenic rats
[9]. A small amount of Spm is known to significantly
reduce the effective concentration of Spd necessary to
support the growth of cells with polyamine deficiency
[10]. It is probable that the same regularity would also
be true in the case of α-methylated derivatives of Spm
and Spd. Therefore, the use of catabolically stable Spm
analogues (e.g., α-MeSpm and α,α'-Me2Spm) that can
fulfill crucial cellular functions of Spm should decrease
the effective concentration of α-MeSpd.
like Spm and Spd, some of these compounds can sup-
port the growth of cells with depleted polyamine pool
[3, 4]. It was shown [3] that α-methylpolyamines are
not the substrates of SSAT, which provides their meta-
bolic stability. At the same time, like Spm and Spd,
1
Corresponding author; phone: +7 (095) 135-6065; fax: +7 (095)
135-1405; e-mail: alexkhom@genome.eimb.relarn.ru
2
Abbreviations: DENSpm, diethyl-nor-spermine; α-MeSpd, α-
methylspermidine; α-MeSpm, α-methylspermine; α,α'-Me -
2
Spm, α,α'-dimethylspermine; NS, o-nitrobenzenesulfonyl; PAO,
polyamine oxidase; Put, putrescine (1,4-diaminobutane); SMO,
spermine oxidase; Spd, spermidine (1,8-diamino-5-azaoctane);
Spm, spermine (1,12-diamino-4,9-diazadodecane); and SSAT,
1
spermine/spermidine N -acetyltransferase (EC 2.3.1.57).
1068-1620/05/3102-0183 © 2005 Pleiades Publishing, Inc.

184
GRIGORENKO et al.
NH
NH2
H2N
NH
NH2
NH
H2N
NH
aMeSpm
a,a'-Me2Spm
NH2
NH2
NH
NH
H2N
H2N
Spd
aMeSpd
The α-methylated analogues of Spm are still quite in particular, they provide the target compounds in
inaccessible despite their biochemical significance. We dozen milligram scale with low overall yields [3].
have recently described a simple synthesis of α-MeSpd
[11]. In this paper, we present convenient schemes for
the preparation of α-MeSpm and α,α'-Me2Spm in
amounts sufficient for studying their metabolism and
biological effects in vitro and in vivo.
Here we describe the building of α-MeSpm back-
bone by a successive elongation of polyamine back-
bone using the alkylation of amino components with
mesylates of the corresponding N-protected amino
alcohols (Scheme 1). The starting N-benzyloxycarbo-
nyl-3-aminobutanol was prepared by the reduction of
commercially available ethyl 3-aminobutyrate with
LiAlH4 in THF followed by the protection of amino
group, using the method previously described for the
synthesis of α-MeSpd [11].
RESULTS AND DISCUSSION
Significant drawbacks are peculiar to the known
preparation methods of α-Me-Spm and α,α'-Me2-Spm;
Cbz
i, ii
iii
NH
CbzNH
CbzNH
CbzNH
OH
CbzNH
N
OH
OH
(I)
(II)
(III)
Cbz
i, iv
v, vi
NH
NH2
N
NH
NH
NH2 · 4HCl
NH2
(IV)
(V)
i, Ms-Cl/Et N/CH Cl ; ii, H N(CH ) OH/THF; iii, Cbz-Cl/H O/NaHCO ; iv, H N(CH ) NH /THF;
3
2
2
2
2 4
2
3
2
2 3
2
v, H /Pd; vi, HCl/MeOH.
2
Scheme 1.
N-Cbz-Alcohol (I) was acylated with methanesulfo- Spm (IV). It was purified by column chromatography
nyl chloride at the first stage of the synthesis. The
resulting mesylate was treated with excess 4-aminobu-
tanol at 50°ë without an additional purification. Crude
amino alcohol (II) was purified by column chromatog-
raphy on silica gel, its secondary amino group was ben-
zyloxycarbonylated, and the completely N-protected
amino alcohol (III) was treated with methanesulfonyl
chloride. The reaction of mesylate of bis-Cbz-amino
alcohol (III) with 10-fold excess of 1,3-diaminopro-
pane in THF at +4°ë [at 20°ë, the number of minor by-
products increased, which made the purification of (IV)
on silica gel. The catalytic hydrogenation of (IV) at
atmospheric pressure over Pd black led to the removal
of Cbz groups, and the target α-MeSpm tetrahydro-
chloride (V) was obtained in 41% total yield from
N-Cbz-3-aminobutanol.
The above linear strategy was ineffective for the
synthesis of α,α'-Me2Spm. The symmetry of the target
molecule allowed us to develop a convergent synthetic
scheme (Scheme 2). Its key step was the alkylation of
N,N'-bis-o-nitrophenylsulfonamide derivative of Put
more laborious] led to bis-N-Cbz-protected α-methyl- (VII) with N-Cbz-3-aminobutyl bromide (VI).
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 31 No. 2 2005

NEW SYNTHESES OF α-METHYL- AND α,α'-DIMETHYLSPERMINE
185
i, ii
CbzNH
Br
CbzNH
H2N
OH
(VI)
(I)
iii
iv, v
NH
CbzNH
NsNH
NH
NHCbz
NH2
NHNs
(VII)
(VIII)
NH
H2N
vi, vii
NH
(IX)
NH2 · 4HCl
i, Ms-Cl/Et N/CH Cl ; ii, LiBr/THF; iii, Ns-Cl/Et N/CH Cl ; iv, (VI)/DMF/K CO ;
3
2
2
3
2
2
2
3
v, PhSH/DMF/K CO ; vi, H /Pd; vii, HCl/MeOH.
2
3
2
Scheme 2.
The treatment of Put with 2.1 equiv of NsCl in the nyl chloride, 4-aminobutanol, and 1,3-diaminopro-
presence of Et3N resulted in bis-sulfonamide (VII), pane from Fluka (Switzerland). N-Benzyloxycarbo-
nyl-3-aminobutanol was prepared according to proce-
dure in [11].
which was difficultly soluble in most of organic sol-
vents except for DMF and DMSO. Bromide (VI) was
prepared by the successive conversion of N-Cbz-ami-
noalcohol (I) into mesylate and, then, without an addi-
tional purification, into the bromide by the treatment
with excess LiBr in THF. After the separation of inor-
ganic salts, (VI) was reacted with sulfonamide (VII) in
the presence of K2CO3, which enabled the alkylation of
both primary amino groups in a high yield. The
attempts to use mesylate or tosylate of (I) for the alky-
lation of sulfonamide (VIII) led only to low yields of
the target product. The selective removal of Ns groups
with the retention of Cbz groups was achieved by the
treatment with a small excess of thiophenol. The result-
ing (VIII) was easily purified by column chromatogra-
phy on silica gel. At the final stage, Cbz groups were
removed by hydrogenation over Pd black at atmo-
spheric pressure and α,α'-Me2Spm tetrahydrochloride
(IX) was isolated with 57% total yield from N-Cbz-3-
aminobutanol.
TLC was carried out on precoated Kieselgel 60 F254
plates (Merck, Germany) using the following systems:
(A) chloroform, (B) 200 : 1 chloroform–methanol, (C)
97 : 3 chloroform–methanol, (D) 9 : 1 chloroform–
methanol, (E) 97 : 3 dioxane–25% ammonia, (F) 9 : 1
dioxane–25% ammonia, and (G) 4 : 2 : 1 : 2 n-butanol–
acetic acid–pyridine–water. Kieselgel (40–63 µm,
Merck, Germany) was used for column chromatogra-
phy. The TLC spots of compounds were visualized in
UV-light and using color reaction with ninhydrin.
1H and 133C NMR spectra were measured on a
Bruker Avance 500 DRX spectrometer (Germany)
using TMS (CDCl3) and sodium 3-(trimethylsilyl)-pro-
panesulfonate (D2O) as internal standards. Chemical
shifts are given in ppm, and constants of spin–spin cou-
pling in Hz.
8-(Benzyloxycarbonyl)amino-5-azanonan-1-ol (II).
A solution of methanesulfonyl chloride (1.24 ml,
16 mmol) in dry dichloromethane (10 ml) was added
for 15 min to a stirred and cooled (0°ë) solution of (I)
(3.34 g, 15 mmol) and triethylamine (2.61 ml,
19 mmol) in dry dichloromethane (40 ml). Stirring was
continued for 1 h at 0°ë and for 1 h at room tempera-
ture. The reaction mixture was poured into 1 M
NaHCO3 (25 ml); the organic layer was separated, suc-
cessively washed with water (10 ml), 0.5 M H2SO4 (3 ×
25 ml), water (10 ml), 1 M NaHCO3 (20 ml), and satu-
rated solution of NaCl (10 ml); dried with MgSO4; and
evaporated in a vacuum. The residue was dissolved in
Thus, the synthetic schemes described in this paper
help prepare α-MeSpm and α,α'-Me2Spm in high total
yields. The purity of the products was more than 99%
as determined by HPLC under the standard conditions
of polyamine analysis [12]. Our synthetic schemes are
also suitable for the preparation of unknown (R)- and
(S)-isomers of α-MeSpm and α,α'-Me2Spm, which
may exhibit different biological activities.
EXPERIMENTAL
Ethyl β-aminobutyrate, 1,4-diaminobutane, and o- THF (20 ml) and 4-aminobutanol (8.9 g, 100 mmol) in
nitrobenzenesulfonyl chloride were from Aldrich THF (10 ml) was added in one portion. The reaction
(United States); benzyl chloroformate, methanesulfo- mixture was kept overnight at 50°ë, THF and 4-ami-
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 31 No. 2 2005

186
GRIGORENKO et al.
nobutanol excess were then distilled off in a vacuum. (10 ml), 0.5 M H2SO4 (3 × 15 ml), water (10 ml), 1 M
The residue was distributed between 1 M NaOH
(20 ml) and chloroform (25 ml); organic layer was sep-
arated, washed with saturated solution of NaCl (3 ml);
dried with MgSO4; and evaporated in a vacuum. The
residue was dissolved in system E and the resulting
solution was separated into three portions. Every por-
tion was purified on a silica gel column (135 g) eluted
with system E. The target fractions were evaporated
and dried in a vacuum over phosphorus pentoxide to get
(II); a viscous oil; yield 3.22 g [73% from (I)]; Rf 0.45
NaHCO3 (10 ml), and saturated solution of NaCl
(10 ml); dried with MgSO4; and evaporated in a vac-
uum. The residue was dissolved in THF (15 ml), cooled
to 0°ë, and 1,3-diaminopropane (6.66 g, 90 mmol) in
THF (5 ml) was added in one portion. The reaction
mixture was kept for 6 h at 0°ë, and overnight at room
temperature. THF and 1,3-diaminopropane excess were
then distilled off in a vacuum. The residue was dis-
solved in 1 M NaOH (15 ml); extracted with dichlo-
romethane (15 ml); and the organic layer was separated
and evaporated in a vacuum. The residue was dissolved
in 94 : 6 dioxane–25% ammonia mixture, and the
resulting solution was separated into two portions.
Each portion was purified on a silica gel column
(135 g) eluted with 94 : 6 dioxane–25% ammonia mix-
ture. The target fractions were evaporated in a vacuum
to dryness, and the residue was dried in a vacuum over
phosphorus pentoxide to get (IV); a viscous oil; yield
3.05 g (70%); Rf 0.25 (E); 1H NMR (CDCl3): 7.34–7.29
(F); 1H NMR (CDCl3): 7.40–7.29 (5 H, m, C6H5), 5.09
(2 H, s, CH2Ph), 5.03 (0.5 H, s, NHCbz), 5.01 (0.5 H, s,
NHCbz), 3.85–3.77 (1 H, m, CH3CH), 3.57 (2 H, t, J
6.32, CH2OH), 2.70–2.55 (4 H, m, CH2NHCH2), 1.76–
1.51 (8 H, m, NH + OH + CH2CH(ëç3)NHCbz +
CH2CH2CH2CH2), and 1.18 (3 H, d, J 6.54, CH3).
N5,N8-Bis-(benzyloxycarbonyl)-8-amino-5-azano-
nanol (III). Benzyl chloroformate (1.55 ml, 11 mmol)
was added in three portions with 15-min intervals to a
cooled (0°ë) and vigorously stirred mixture of (II) (10 H, m, C6H5), 5.10 (2 H, s, CH2Ph), 5.07 (2 H, s,
(2.94 g, 10 mmol), THF (10 ml), water (5 ml),
NaHCO3 (0.84 g, 10 mmol), and 10 M NaOH (0.6 ml,
6 mmol). The stirring was continued for 1 h at 0°ë and
then for 3 h at room temperature. The aqueous layer
was separated, extracted with chloroform (15 ml), and
the combined organic phase was evaporated to dryness
in a vacuum. The residue was dissolved in chloroform
CH2Ph), 4.79 (1 H, s, NHCbz), 3.35–3.20 (5 H, m,
CH3CH + CH2N(Cbz)CH2), 2.73 (2 H, t, J 6.75,
CH2NH2), 2.66–2.53 (4 H, m, CH2NHCH2), 1.70–1.35
(11 H, m, NH + NH2 + CH2CH2NH + CbzNHCHCH2 +
CH2CH2CH2CH2), and 1.16–1.10 (3 H, m, CH3); 13C
NMR (CDCl3): 155.85, 136.88, 136.70, 128.48,
(30 ml); the solution was washed with 0.5 M H2SO4 (2 × 128.02, 127.92, 127.79, 67.07, 66.96, 66.48, 49.63,
47.85, 47.62, 47.17, 45.37, 44.77, 44.32, 40.54, 36.20,
35.06, 33.77, 27.25, 26.48, 25.95, and 21.33.
10 ml), water (15 ml), and 1 M NaHCO3 (2 × 10 ml);
dried with MgSO4, and evaporated in a vacuum. The
residue was dissolved in chloroform, and the resulting
solution was separated into two portions. Every portion
was purified on a silica gel column (120 g) eluted first
with a 200 : 1 chloroform–methanol and then with a
95 : 5 chloroform–methanol mixtures to get (III); a col-
1,12-Diamino-4,9-diazatridecane
tetrahydro-
chloride (a-MeSpm) (V). A suspension of Pd black in
methanol (~0.9 ml) was added to a solution of (IV)
(3.0 g, 6.2 mmol) in a 1 : 1 AcOH–MeOH mixture
(30 ml), and hydrogenation was carried out at atmo-
spheric pressure. The catalyst was filtered off, washed
with methanol (20 ml), and the combined filtrates were
evaporated to dryness in a vacuum. The residue was
dissolved in methanol, diluted with 5 M HCl (7.4 ml),
and the resulting solution was evaporated to dryness in
a vacuum. The residue was recrystallized from a water–
MeOH–EtOH mixture and dried in a vacuum over
P2O5/KOH to give (V) tetrahydrochloride as colorless
crystals; yield 1.97 g (88%); mp 250–251°ë (dec.) {lit.
[3]: mp 247°ë (dec.) for lyophilized powder}; Rf 0.13
1
orless viscous oil; yield 3.94 g (92%); Rf 0.27 (C); H
NMR (CDCl3): 7.34–7.27 (10 H, m, C6H5), 5.11 (2 H,
s, CH2Ph), 5.07 (2 H, s, CH2Ph), 4.96 (0.5 H, s,
NHCbz), 4.57 (0.5 H, s, NHCbz), 3.73–3.55 (3 H, m,
CH3CH
+
CH2OH), 3.30–3.18 (4 H, m,
CH2N(Cbz)CH2), 1.75–1.45 (7 H, m, OH
+
CH2CH(ëç3)NHCbz + CH2CH2CH2CH2), and 1.17–
1.10 (3 H, m, CH3); 13C NMR (CDCl3): 156.16, 155.90,
136.82, 128.52, 128.07, 128.00, 127.85, 67.07, 66.59,
62.33, 47.44, 45.41, 44.78, 44.34, 36.22, 35.13, 29.66,
25.02, 24.70, and 21.32.
1
(G); H NMR (D2O): 3.51 (1 H, q, J 6.83, CH3CH),
N9,N12-Bis-(benzyloxycarbonyl)-1,12-diamino-4,9-
diazatridecane (IV). A solution of MsCl (0.77 ml,
10 mmol) in dry dichloromethane (5 ml) was added for
10 min to a stirred and cooled (0°ë) solution of (III)
(3.85 g, 9 mmol) and Et3N (1.53 ml, 11 mmol) in dry
dichloromethane (20 ml). The stirring was continued
for 1 h at 0°ë and for 30 min at room temperature. The
reaction mixture was poured into 1 M NaHCO3 (20 ml);
3.24–3.11 (10 H, m, CH2NH + CH2NH2), 2.18–2.08
(3 H, m, NHCHCH2 + CH2CH2NH2), 2.04–1.96 (1 H, m,
NHCHCH2), 1.81 (4 H, m, CH2CH2CH2CH2), and 1.36
(3 H, d, J 6.60, CH3); 13C NMR (CDCl3): 47.07, 45.41,
44.62, 44.00, 36.67, 30.47, 23.79, 22.83, 17.58, and
17.34. Found, %: C 36.03, H 9.00, N 15.04.
C11H32N4Cl4 · 0.25H2O. Calculated, %: C 36.03, H
the organic layer was separated, washed with water 8.93, N 15.28.
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 31 No. 2 2005

NEW SYNTHESES OF α-METHYL- AND α,α'-DIMETHYLSPERMINE
187
N3-(Benzyloxycarbonyl)-amino-1-bromobutane
(VI). A solution of MsCl (1.16 ml, 15 mmol) in dry
dichloromethane (10 ml) was added for 15 min to a
stirred and cooled (0°C) solution of (I) (3.01 g,
form was distilled off in a vacuum, and the residue was
dissolved in 1,4-dioxane (10 ml) and purified on a silica
gel column (130 g) eluted with system (E). After evap-
oration in a vacuum and drying the residue in a vacuum
13.5 mmol) and Et3N (2.52 ml, 18.1 mmol) in dry over phosphorus pentoxide, (VIII) was obtained as col-
orless crystals; yield 2.3 g [70% from (VI)]; Rf 0.46 (E);
mp 107–108°ë; 1H NMR (CDCl3): 7.36–7.26 (10 H, m,
C6H5); 5.51 (2 H, bs, NHCbz), 5.08 (4 H, s, CH2Ph),
3.85–3.74 (2 H, m, CH3CH), 2.73–2.65 (2 H, m,
CH2NH), 2.64–2.54 (2 H, m, CH2NH); 2.54–2.45 (4 H,
m, CH2NH), 1.72–1.40 (8 H, m, CH2CHNHCbz +
CH2CHNHCbz + CH2CH2CH2CH2), and 1.16 (6 H, d,
dichloromethane (30 ml). Stirring was continued for
30 min at 0°C and for 30 min at room temperature. The
reaction mixture was poured into 1 M NaHCO3 (30 ml);
the organic layer was separated, washed with water
(10 ml), 0.5 M H2SO4 (3 × 15 ml), water (10 ml), and
1 M NaHCO3 (10 ml); dried with MgSO4; and evapo-
rated in a vacuum. The residue was dissolved in THF
(5 ml), and LiBr (3.5 g, 40 mmol) in THF (20 ml) was
added in one portion. The reaction mixture was stirred
overnight at room temperature and precipitated with
chloroform (30 ml). The precipitate was filtered off,
and the filtrate was evaporated in a vacuum. The residue
was treated with chloroform (50 ml), washed with
water (2 × 30 ml), 1 M NaHCO3 (2 × 15 ml), dried with
MgSO4, evaporated in a vacuum and, then, dried in a
vacuum over P2O5 /KOH to give (VI); a viscous oil;
J 6.48, CH3); 13C NMR (CDCl3): 156.03, 136.86,
128.53, 128.05, 67.14, 66.41, 49.82, 46.45, 46.01,
36.66, 27.87, and 21.28.
2,13-Diamino-5,10-diazatetradecane tetrahydro-
chloride (a,a'-Me2Spm) (IX). A suspension of Pd
black in methanol (~0.8 ml) was added to a solution of
(IV) (2.3 g, 4.6 mmol) in a 1 : 1 AcOH–MeOH mixture
(30 ml), and hydrogenation was carried out at atmo-
spheric pressure until the evolution of carbon dioxide
ceased (for approximately 3 h). Pd black was filtered
off, washed with MeOH (15 ml), and the combined fil-
trates were evaporated to dryness in a vacuum. The res-
idue was dissolved in methanol (15 ml), diluted with
5 M HCl (6.0 ml), and the resulting solution was evap-
orated to dryness in a vacuum. The residue was recrys-
tallized from a water–MeOH–EtOH mixture, and the
resulting colorless crystals of (IX) were dried in a vac-
uum over P2O5/KOH; yield 1.47 g (85%), mp 224–
225°ë (dec.) {lit. [3]: mp 180°ë (dec.) for a lyophilized
powder}; Rf 0.13 (G); 1H NMR (D2O): 3.55–3.46 (2 H,
m, CH3CH), 3.23–3.18 (4 H, t, J 6.77, CH2NH), 3.17–
3.11 (4 H, m, CH2NH), 2.18–2.09 (2 H, m,
CH2CHNH2), 2.04–1.93 (2 H, m, CH2CHNH2), 1.84–
1.75 (4 H, m, CH2CH2CH2CH2), and 1.35 (6 H, d,
1
yield 3.78 g (95%); Rf 0.61 (A); H NMR (CDCl3):
7.37–7.26 (5 H, m, C6H5), 7.11 (1 H, bd, NHCbz), 5.03
(2 H, s, CH2Ph), 3.78–3.72 (1 H, m, CH3CH), 3.46–
3.41 (2 H, t, J 7.38, CH2Br), 2.10–1.99 (1 H, m,
CH3CHCH2), 1.95–1.85 (1 H, m, CH3CHCH2), and
1.12 (3 H, d, J 6.63, CH3). It was used without further
purification.
Bis-N1,N4-o-nitrobenzenesulfonyl-1,4-diaminobu-
tane (VII). A solution of o-nitrobenzenesulfonyl chlo-
ride (6.05 g, 27.3 mmol) in dry dichloromethane
(30 ml) was added for 40 min to a cooled (0°ë) and
vigorously stirred solution of freshly distilled 1,4-
diaminobutane (1.1 g, 12.5 mmol) and Et3N (5.2 ml,
37.5 mmol) in dry dichloromethane (50 ml). Stirring
was continued for 1 h at 0°ë and 3 h at room tempera-
ture. The precipitate was filtered off, washed with
MeOH (3 × 15 ml), chloroform (3 × 10 ml), and dried
over P2O5 to give (VII), yield 5.3 g (91%) as a pale-yel-
J 6.63, CH3); 13C NMR (D2O): 49.86, 48.14, 46.78,
33.27, 25.64, and 20.09. Found, %: C 38.08, H 9.24, N
14.67. C12H34N4Cl4. Calculated, %: C 38.31, H 9.11, N
14.89.
1
low crystals; Rf 0.55 (B); mp 185–186°C; H NMR
(DMSO-d6): 7.98–7.91 (4 H, m, C6H4), 7.86–7.81 (4 H,
m, C6H4), 2.87–2.82 (4 H, bt, CH2NHNs), 1.44-1.37
(4 H, m, CH2CH2CH2CH2).
Bis-N2,N13-(benzyloxycarbonyl)-2,13-diamino-5,10-
diazatetradecane (VIII). A suspension of bromide
(VI) (3.78 g, 13.2 mmol), bissulfonamide (VII) (2.16 g,
4.7 mmol), and K2CO3 (4.30 g, 31 mmol) in anhydrous
DMF (30 ml) was stirred for 48 h at room temperature.
DMF (10 ml), K2CO3 (3.7 g, 27 mmol), and PhSH
(1.4 ml, 13.5 mmol) were then added to the reaction
mixture, and stirring was continued for 12 h at room
temperature. The reaction mixture was evaporated to
dryness in a vacuum. The residue was treated with chlo-
roform (50 ml); the precipitate was filtered off and
washed with chloroform (3 × 10 ml); and the combined
filtrates were washed with water (2 × 30 ml). Chloro-
ACKNOWLEDGMENTS
This work was supported by the Academy of Sci-
ences of Finland and by the Russian Foundation for
Basic Research, project no. 03-04-49080.
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