Synthesis of novel deuterium labelled derivatives of N-alkylated polyamines

Article abstract of DOI:10.1016/j.tet.2008.10.071

Novel di-, tetra- and octadeuterated derivatives of mono-N-alkylated diaminopropanes, spermidines, spermines, symmetrically bis-N-alkylated spermines and unsymmetrically bis-N-alkylated spermines were synthesized. Deuterium labels were introduced into the RHNCH₂CH₂CN intermediate either by exchanging the protons next to the nitrile group under basic conditions with D₂O-EtOD mixture or/and by reducing the nitrile group to a CD₂-NH₂ fragment with LiAlD₄.

Full text of DOI:10.1016/j.tet.2008.10.071

Tetrahedron 65 (2009) 547–562
Contents lists available at ScienceDirect
Tetrahedron
journal homepage: www.elsevier.com/locate/tet
Synthesis of novel deuterium labelled derivatives of N-alkylated polyamines
,
b
c
d
a
Merja R. Ha¨kkinen a , Tuomo A. Keina¨nen , Alex R. Khomutov , Seppo Auriola , Janne Weisell ,
*
Leena Alhonen b, Juhani Ja¨nne b, Jouko Vepsa¨la¨inen a
a Department of Biosciences, Laboratory of Chemistry, Biocenter Kuopio, University of Kuopio, PO Box 1627, FI-70211 Kuopio, Finland
b Department of Biotechnology and Molecular Medicine, A.I. Virtanen Institute for Molecular Sciences, Biocenter Kuopio, University of Kuopio,
PO Box 1627, FI-70211 Kuopio, Finland
c Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Street 32, Moscow 119991, Russia
d Department of Pharmaceutical Chemistry, Biocenter Kuopio, University of Kuopio, PO Box 1627, FI-70211 Kuopio, Finland
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 9 May 2008
Received in revised form 10 October 2008
Accepted 23 October 2008
Available online 26 October 2008
Novel di-, tetra- and octadeuterated derivatives of mono-N-alkylated diaminopropanes, spermidines,
spermines, symmetrically bis-N-alkylated spermines and unsymmetrically bis-N-alkylated spermines
were synthesized. Deuterium labels were introduced into the RHNCH2CH2CN intermediate either by
exchanging the protons next to the nitrile group under basic conditions with D2O–EtOD mixture or/and
by reducing the nitrile group to a CD2–NH2 fragment with LiAlD4.
Ó 2008 Elsevier Ltd. All rights reserved.
Keywords:
Polyamines
Deuterated polyamines
Deuterated
N-Alkylated polyamine analogues
NH2
H2N
N
H
1. Introduction
SPD
SPM
Biogenic polyamines (PAs), spermidine (SPD), spermine
(SPM) and their diamine precursor putrescine (butane-1,4-di-
amine) are small organic bases, which are widely distributed in
nearly every prokaryotic and eukaryotic cell type, and are es-
sential for cell growth, division and differentiation. In mam-
malian cells, the PAs are present at micro- to millimolar
concentrations, and their intracellular levels are known to be
strictly regulated by several enzymes and the cell membrane
transport system.1–3 Natural PAs and their analogues have been
investigated as chemopreventative and antiparasitic agents,
NMDA receptor modulators, polyamine-based venoms, metal
chelators and as potential carriers for drug delivery.4–6 It is
known that relatively modest structural modifications in the PA
backbone or incorporation of terminal N-alkyl substituent(s)
may evoke significant differences in their chemical and thera-
peutic behaviour.5–10
H
N
NH2
H2N
N
H
Metabolic studies play an important role in drug discovery,
development, testing and approval,11 and represent the basic
knowledge needed prior to clinical drug use by providing
information on the drug safety and efficacy. This may prevent un-
necessary expense in drug development as unsuitable projects can
be interrupted as quickly as possible. In addition, metabolic studies
may also identify novel possible targets for therapeutic
intervention and new avenues for research.5,6
Despite intensive investigations into the biochemistry and
molecular biology of PAs, there is relatively sparse data describing
details of the interaction between PAs with macromolecules and
receptors at the molecular level. In other words, the actual in-
tracellular roles of PAs and the specificity of their interactions with
receptor molecules are still largely unknown.1,3,6,9 One reason for
this problem is that it is biochemically challenging due to the fact
that different PAs compete for the same binding site with varying
degrees of specificity for their many predicted target molecules,
* Corresponding author. Tel.: þ358 40 3553987; fax: þ358 17 163259.
E-mail address: merja.hakkinen@uku.fi (M.R. Ha¨kkinen).
0040-4020/$ – see front matter Ó 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.tet.2008.10.071

548
M.R. Ha¨kkinen et al. / Tetrahedron 65 (2009) 547–562
and there can also be competition with divalent metal ions or
other polyamine mimetics in the study systems. Moreover, the
cellular distribution and compartmentalization of polyamines is
not understood in detail. Furthermore, in vivo studies are ham-
pered due to the metabolism of polyamines and their structural
analogues. Thus, novel tools are required for in depth studies to
identify key cellular targets as well as understanding details of
the substrate properties of polyamine(analogue) metabolizing
enzymes.
Today, liquid chromatography (LC) coupled to tandem mass
spectrometry (MS/MS) represents a powerful tool for metabolite
profiling studies since it can perform metabolite identification,
as well as structural characterization and quantitation of the
metabolites.12 Recently, we have developed a novel method in
which PAs, and also their biologically active derivatives and
metabolites, are separated and quantitatively analyzed by
LC–MS/MS using deuterium labelled compounds as internal
standards.13,14
2. Results and discussion
2.1. Chemistry
The most straightforward method to synthesize deuterium la-
belled molecules is to use commercially available labelled building
blocks, but this approach is often limited due to the low availability
and also the high cost of these compounds. It is possible to attempt
exchange reactions adjacent to an activation group accelerated by
various catalytic methods, and by using deuterated solvents as the
labelling source.22,25 However, the most common and a selective
deuteration method is reduction with deuterium gas or deuterated
solvents with the appropriate catalyst, or with metal deu-
terides.20,21,26,27 Some possible approaches, which could be used in
the synthesis of deuterium labelled polyamines are summarized in
Scheme 1.
O
O
O
CN
or
X
To achieve acceptable reproducibility and reliability, internal
standards are used to correct for the variations in sample prep-
aration and to compensate for the variability in MS detection.
Stable isotopically labelled analogues are the most appropriate
internal standards in quantitative LC–MS/MS assays, since virtu-
ally the only distinguishing feature from the analyte is the dif-
ference in the m/z ratio of the ions being studied. However, their
use has been hampered due to the time-consuming and expen-
sive syntheses involved in their production.12,15 Deuterium is the
most widely accessible and the least expensive stable isotope for
use in the preparation of labelled internal standards. It is also
often synthetically straightforward to incorporate more than one
deuterium atom into the molecule, which may be necessary to
sufficiently distinguish the labelled internal standards from the
isotope peaks of the unlabelled analytes. In addition, deuterium
labelled compounds are invaluable tools in studying the mecha-
nisms of enzymes and the site of biotransformation, establishing
the mechanism of organic reactions, and they are also useful
in structural determinations, for example, in MS and NMR
analyses.16,17
Various protocols have been developed for the synthesis of PA
analogues and conjugates,4,6,7,18 but the synthetic strategies to
obtain deuterium labelled polyamines have been much more lim-
ited, and there is no general method to prepare N-alkylated PA
derivatives labelled with deuterium.19–22 Previously, only putres-
cine-d4 (1,1,4,4-2H4-butane-1,4-diamine),21 putrescine-d8,20,23
cadaverine-d4 (1,1,5,5-2H4-pentane-1,4-diamine),21 1,1,3,3-2H4-
propane-1,3-diamine22 and 1,1,2,3-2H4-propane-1,3-diamine20
have been used to prepare deuterated spermine, spermidine, N1-
acetylspermine and N1-acetylspermidine.19–21,24 However, the
selective derivatization of diamines is not straightforward, even in
the simplest cases for the synthesis of N-monosubstituted alkane-
diamines,22 and it is especially complicated with non-symmetrical
diamines.
several
steps
D2O/D+
b,c=CHD
b=CD2
O
e
LiAlD4
a
a
c
a
b
N
N
R
N
e
b
N
e
b
a
R
a=CD2
D2O/D+
[D]
a=CD2
CN
e=D
N
H
Scheme 1. Possible retrosynthetic strategies to prepare deuterated polyamine
derivatives.
Our synthetic strategies to deuterated PA analogues 1–4 are
based on the methods described above. As shown in Scheme 2, the
starting compound was acrylonitrile (5), since it allows not only the
introduction of the desired N-alkyl group to the double bond, but
also the deuterium labelling of either X or Y positions, or both. The
key intermediate 1, after sufficient protection, can undergo chain
elongation to yield the corresponding deuterium labelled SPD and
SPM derivatives. The structures of prepared target compounds 1–4,
their overall yields and the isotopic purities of the deuterated
compounds are summarized in Tables 1–3.
X
X
i
ii
R
R
CN
CN
N
H
N
H
NH2
Y
Y
5
6a-d
iii
1a-k
ii
CN
R
N
H
6e,f
D
D
Scheme 2. (i) R–NH2 (R¼Et, iPr, c-Hex, Bn), MeOH, reflux; (ii) Method A: LiAlH4 or
LiAlD4, THF, reflux, Method B: H2/PtO2, EtOH, HCl; (iii) 0.1 M NaOD, D2O, EtOD; X and
Y¼H or D.
The aim of this study was to develop a simple and straightfor-
ward method to synthesize N-alkylated polyamines 1–4, in which
R¼H, Et, iPr, Bn or c-Hex, and X, Y, X1 and Y1 are deuterium labelling
positions, in order to use these compounds as standards in quan-
titative analysis of polyamines by LC–MS/MS,13,14 as well as bio-
chemical probes to study the metabolism of PAs.
Preparation of PA 1–4 was started from acrylonitrile (5) and the
appropriate N-alkylamine using the Michael addition to obtain the
desired N-alkylamino-propionitriles 6a–d with 79–91% yields.28 In
H
N
H
N
H
N
X
X
Y1 Y1
X
X
X
X
X
X
Y1 Y1
R
R
R
NH2
R
NH2
N
H
N
H
R1
N
H
NH2
N
H
N
H
N
H
N
H
Y
Y
X1 X1
Y
Y
Y
Y
Y
Y
X1 X1
4
1
2
3

M.R. Ha¨kkinen et al. / Tetrahedron 65 (2009) 547–562
549
Table 1
a
-position. The deuteration stage achieved >97% for 6e,f after the
Synthesized DAP derivatives, their overall yields and the isotopic purities of the
deuterium labelled compounds
exchange reaction was repeated twice with a mixture of 0.1 M
NaOD in D2O and EtOD by using dry diethylether as the extraction
solvent. Ethanol is of crucial importance for this reaction, since in
its absence after the first cycle, only about 82–84% hydrogen was
exchanged, whereas with ethanol, the deuterium stages were 92–
95%. Moreover, chloroform was a poor extraction solvent, since
Compound
Overall yield %a
Intermediate Method
Isotopic
purity %
DAP derivatives
R
X
Y
compound
A
B
C
1a
1b
1c
1d
1e
1f
1g
1h
1i
Et
Et
Et
Et
H
D
H
D
H
D
D
H
D
H
D
D
H
H
D
D
H
H
D
H
H
H
H
H
6a
6a
6e
6e
6d
6d
6f
6b
6b
6c
6c
1f
39
43
18
20
62
70
49
46
47
56
60
54
73
d
99
94
92
d
98
98
d
99
d
98
98
both nitrogen deuteriums and a-deuteriums to nitrile were con-
verted back to hydrogens to some extent, even when the traces of
water and ethanol (as a stabilizer in chloroform) were removed
with P2O5.30
Bn
Bn
Bn
iPr
56
The obtained nitriles 6a–f were reduced to the corresponding
amines 1a–k either with LiAlD4 (method A, 43–88% yields) or with
PtO2 catalysed hydrogenation (method B, 80–84% yields). However,
the latter method is not suitable for benzyl derivatives 1e–g, and as
an example, compound 1e was prepared from the commercially
available Boc-protected amine 7 and benzaldehyde via imine 8
using reductive amination (method C) as shown in Scheme 3. Also
the chain elongation unit 10, which was used as a building block to
the SPM derivatives 3a–g, was prepared from 7 via protected 9.
Previously, 10 (X¼Br) was prepared from 9 and 1,4-dibromobutane
but we used 1-bromo-4-chlorobutane as an alkylating agent to
avoid dimerization.31
66
72
iPr
1j
1k
1l
c-Hex
c-Hex
H
a
Method A and method B, starting from alkylamine and acrylonitrile. Method A
with LiAlH4 or LiAlD4 and Method B with H2/PtO2. Method C, starting from aldehyde.
Yields are for recrystallized compounds and are not optimized.
Table 2
Synthesized SPD derivatives, their overall yields and the isotopic purities of the
deuterium labelled compounds
Compound
Overall yield %a
Intermediate Method
Isotopic
purity %
Ph
SPD derivatives
R
X
Y
Boc
Boc
H
ii
i
compound
N
N
H
H2N
N
H
1e
A
B
C
7
8
2a
2b
2c
2d
2e
2f
Et
Et
Et
Bn
Bn
iPr
c-Hex
H
H
D
D
H
D
H
H
D
H
H
D
H
H
H
H
H
12a
12b
12c
12d
12e
12f
12g
2e
25
21
15
31
39
14
19
21
46
d
99
93
d
97
d
d
99
iii
28
iv
Ns
X
Boc
N
H
N
N
Ns
N
H
20
25
Boc
9
2g
2h
10a, X=Cl
10b, X=I
v
a
Method A and method B, starting from alkylamine and acrylonitrile. Method C,
starting from aldehyde. Yields are for recrystallized compounds and are not
optimized.
Scheme 3. (i) PhCHO, 3 Å molecular sieves, Et2O; (ii) (1) NaBH4, EtOH; (2) HCl, EtOH;
(iii) NsCl, Et3N, CH2Cl2; (iv) Br(CH2)4Cl, K2CO3, DMF; (v) NaI, acetone, reflux.
Mono-N-alkylated SPD analogues were selectively prepared
from propanediamines 1a–k after protection as the o-nitro-
phenylsulfonyl (Ns) derivatives 11a–k (intermediate structures in
Tables 7–10), since the only remaining NH group was readily
alkylated with N-(4-iodobutyl)phthalimide32 to 12a–k under basic
conditions with 81–95% yields as shown in Scheme 4. It was no-
table that the protection of the sterically hindered N-isopropyl and
N-cyclohexyl derivatives to nosylated 11g–h required more robust
reaction conditions, chromatographic purification and yields (40–
48%) were only half of that of those obtained with the ethyl and
benzyl derivatives. The target SPD hydrochlorides 2a–g were
obtained from 12a–k after deprotection and recrystallization from
ethanol–water–EtOAc with 51–80% yields.
Mono-N-alkylated SPM analogues 3a–f were also prepared from
nosylated 11 via 13a–f using 10b (see Scheme 3) as an alkylating
agent with 35–60% yields following the strategy having been earlier
developed for spermidines.32,33 Using the same approach and with
1,4-diiodobutane as the alkylating agent, the corresponding
symmetric di-N,N-alkyl SPMs 4a–d were obtained via 14a–d with
32–69% yields. Surprisingly, the purification of 14a–c was
straightforward, since after addition of EtOAc and water to the re-
action mixtures, the products crystallized out. Thus, chromato-
graphic purification was needed only for the compound 14d.
Using this strategy it is also possible to prepare non-symmetric
N,N0-dialkyl SPMs. In this case, as shown in Scheme 4, we used 11a
(R¼Et, X¼Y¼H) or 11c (R¼Et, X¼Y¼D) as starting materials to
prepare alkylating agents 15c and 15d, respectively. The com-
pounds 11d (R¼Bn, X¼Y¼H) or 11f (R¼Bn, X¼Y¼D) were selected
the next step, it was possible to exchange –CH2CN protons with
deuterium or to reduce the nitrile group to the CD2–NH2 or CH2–
NH2 unit. The base catalyzed hydrogen exchange reactions of the
simple cyanoalkanes are much faster than the corresponding hy-
drolysis to carboxylate ions,29 which enables labelling at the
Table 3
Synthesized SPM derivatives, their overall yields and the isotopic purities of the
deuterium labelled compounds
Compound
Overall yield %a Isotopic
purity %
SPM
R
R1
X
Y
X1 Y1 Intermediate Method
derivatives
compound
A
B
C
3a
3b
3g
3c
3d
3e
3f
4a
4b
4c
4d
4e
4f
Et
Et
Et
Bn
Bn
iPr
c-Hex
Et
H
H
H
H
H
H
H
Et
Et
iPr
H
D
H
H
D
H
H
H
D
H
H
H
H
D
D
H
H
H
H
D
H
H
H
H
H
H
H
H
D
D
H
H
D
H
H
H
H
H
D
H
H
H
D
D
H
H
H
D
H
H
H
H
H
H
H
H
H
D
D
H
13a
13b
4f
13
15
16
28
27
11
12
16
14
10
18
21
20
10
22
25
d
97
93
d
92
d
d
d
97
d
d
d
92
87
92
13c
13d
13e
13f
14a
14b
14c
14d
14e
14f
14g
3d
26
15
16
30
Et
iPr
15
24
40
c-Hex c-Hex
Et
Et
Et
H
Bn
Bn
Bn
H
4g
4h
a
Method A and method B, starting from alkylamine and acrylonitrile. Method C,
starting from aldehyde. Yields are for recrystallized compounds and are not
optimized.

550
M.R. Ha¨kkinen et al. / Tetrahedron 65 (2009) 547–562
O
iii
X
X
2a-g
R
R
R
N
N
Ns
N
Ns
ii
Y
Y
Y
O
X
X
X X
12a-g
i
R
R
H
N
H
NH2
N
Ns
N
iv
Ns
N
H
N
v
X
X
Y
Y
Y Y
Ns
3a-f
10b
N
Ns
N
Ns
Boc
R1
1a-k
11a-h
Y
13a-f
vii
vi
ix
Ns
N
Ns
N
X
X
X
X
Y1 Y1
v
Z
4a-g
N
Ns
N
Ns
N
Ns
N
Ns
Y
Y
Y
Y
X1 X1
14a-g
15a-b, Z=Cl
15c-d, Z=I
viii
Scheme 4. (i) NsCl, Et3N, CH2Cl2; (ii) PhtN(CH2)4I,32 K2CO3, DMF; (iii) (1) PhSH, K2CO3, DMF; (2) N2H4$H2O, EtOH, reflux; (3) HCl, 1,4-dioxane; (iv) 10b, K2CO3, DMF; (v) (1) PhSH,
K2CO3, DMF; (2) HCl, 1,4-dioxane; (vi) I(CH2)4I, K2CO3, DMF; (vii) Br(CH2)4Cl, K2CO3, DMF; (viii) NaI, acetone, reflux; (ix) 11, K2CO3, DMF.
as the precursors of another three-carbon fragment of the SPM
Table 4
13C NMR chemical shifts for DAP derivatives 1a–la,b,c
molecule. The use of these benzyl derivatives not only allowed the
preparation of mixed N-ethyl-N-benzyl SPMs 4e–g, but also after
the removal of benzyl group by catalytic hydrogenation, it proved
possible to obtain SPM derivatives labelled at the non-alkylated
terminus (see compound 3g in Table 3). The same strategy was used
to prepare deuterated propanediamine-d2 1l, spermidine-d2 2h and
spermine-d4 4h starting from 1f, 2e and 3d, respectively, as
depicted in Scheme 5.
Compd
C3
R
C1
N
H
C2 NH2
R
C-1
C-2
1JCD
C-3
1JCD
1a
1b
1c
1d
1e
1f
1g
1h
1i
Et
Et
Et
Et
46.8
46.8
46.7
46.7
46.8
46.8
46.6
44.5
44.5
44.2
44.1
39.4
26.6
26.4
26.0*
25.8*
26.6
26.5
25.7*
26.9
26.6
26.8
26.6
27.5
39.5
38.9*
39.3
38.8*
39.4
39.0*
38.8*
39.5
39.0*
39.5
39.0*
38.9*
22 Hz
22 Hz
20 Hz
20 Hz
Bn
Bn
Bn
iPr
22 Hz
22 Hz
D
D
20 Hz
NH2
H2N
N
iPr
22 Hz
H
2h
1j
1k
1l
c-Hex
c-Hex
H
i
22 Hz
22 Hz
X
X
D
D
2e
i
a
b
c
NMR solvent was D2O with TSP and products were measured as their HCl salts.
13C NMR chemical shifts for R are presented in Section 4.
Shifts marked with * are for CD2 carbons, which were observed as 1:2:3:2:1
1f
Ph
N
H
N
H
H2N
NH2
3d
Y
Y
1l
i
quintets.
H
N
D
D
D
signals were observed as shown in Figure 1. When the protons are
replaced by deuterium the relaxation time (T1) for the deuterated
carbon may be more than an order of magnitude greater than that
of the comparable protonated carbon. This may mean that with
normal recycle delay for carbon measurements (D1¼3 s) deuter-
ated carbons may either not show up or appear with much reduced
intensity. This was the reason to collect 13C NMR data over a long
period of time with a longer recycle delay (D1¼10 s). In addition,
the intensity of the carbon signal will be split into a multiplet due to
13C–2H J coupling depending on how many deuterons are attached
to the carbon (1:1:1 triplet for CD and 1:2:3:2:1 quintet for CD2).
Measured 13C–2H J couplings (1JCDz19–22 Hz) for quintets are
NH2
H2N
N
H
D
4h
Scheme 5. (i) H2/10% Pd–C, EtOH–H2O.
2.2. NMR, MS and IR studies
All of the compounds were identified by 1H and 13C NMR
spectroscopy. The complicated 13C NMR spectra were assigned
based on 2D techniques and via the information obtained from
deuterated analogues. All the backbone carbons are quoted in Ta-
bles 4–6 (target compounds 1–4) and in Tables 7–10 (in-
termediates). In the experimental part and in Tables 4–10, the
backbone carbons are assigned with a number and N-alkyl
substituents by using Greek letters as shown below.
Table 5
13C NMR chemical shifts for SPD derivatives 2a–ha,b,c
Compd
C7
R
C1
C3 C5
NH2
N
H
C2
N
H
C6
C8
C-5
50.0 25.7 26.9 41.7
δ
1
3
5
R
C-1
C-2
1JCD
C-3
1JCD
C-6
C-7
C-8
N
N
γ
N
N
α
β
2a
2b
2c
2d
2e
2f
Et
Et
Et
Bn
Bn
iPr
46.8 25.7
46.7 25.4
47.4
8
12
46.8* 22 Hz 49.9 25.6 26.8 41.7
46.6 24.7* 20 Hz 46.6* 22 Hz 49.9 25.6 26.7 41.7
46.8 25.6
46.7 25.3
44.5 25.8
47.4
50.0 25.7 26.8 41.7
During the syntheses, the overall deuterium contents were es-
timated from the 1H NMR spectra and localizations were verified
from the 13C NMR spectra. The presence of deuterium atoms in the
target compounds were identified without doubt, since in the 1H
NMR spectra the exchanged proton signals disappeared, remaining
coupling patterns were simplified and in the 13C NMR spectra, the
low intensity CD2 quintets closely upfield to the corresponding CH2
46.8* 22 Hz 49.8 25.5 26.7 41.7
47.4
47.5
50.0 25.6 26.8 41.7
50.0 25.7 26.8 41.7
2g
2h
c-Hex 44.2 25.8
39.4 26.4
H
46.8* 22 Hz 49.9 25.6 26.7 41.7
a
b
c
NMR solvent was D2O with TSP and products were measured as their HCl salts.
13C NMR chemical shifts for R are presented in Section 4.
Shifts marked with * are for CD2 carbons, which were observed as quintets.

M.R. Ha¨kkinen et al. / Tetrahedron 65 (2009) 547–562
551
Table 6
13C NMR chemical shifts for SPM derivatives 3a–g and 4a–ha,b,c
Compd
H
N
H
N
C11
R
C1
C3 C5
C7
C12
C-7
N
H
C2
N
H
C6
C8 C10
R1
C-8
R
R1
C-1
C-2
1JCD
C-3
1JCD
C-5
C-6
C-10
1JCD
C-11
1JCD
C-12
3a
3b
3g
3c
3d
3e
3f
4a
4b
4c
4d
4e
4f
Et
Et
Et
Bn
Bn
iPr
c-Hex
Et
H
H
H
H
H
H
H
Et
Et
iPr
c-Hex
Et
Et
Et
H
46.7
46.7
46.7
46.7
46.6
44.5
44.1
46.7
46.7
44.5
44.1
46.7
46.6
46.6
39.2
25.5
25.4
25.5
25.5
24.7*
25.8
25.8
25.6
25.4
25.8
25.8
25.5
24.7*
24.7*
25.8*
47.4
46.8*
47.3
47.4
46.6*
47.4
47.4
47.4
46.8*
47.4
47.4
49.9
49.8
49.8
49.9
49.8
49.9
49.9
49.9
49.8
49.9
49.9
49.9
49.8
49.8
49.8
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
25.6
49.9
49.9
49.8
49.9
49.9
49.9
49.9
49.9
49.8
49.9
49.9
49.9
49.9
49.8
49.8
47.4
47.4
46.7*
47.4
47.4
47.4
47.4
47.4
46.8*
47.4
47.4
47.4
47.4
46.6*
47.4
26.6
26.6
25.8*
26.6
26.6
26.6
26.6
25.6
25.4
25.8
25.8
25.6
25.5
24.7*
26.6
39.5
39.4
39.2
39.4
39.4
39.4
39.4
46.7
46.7
44.5
44.1
46.7
46.7
46.5
39.4
22 Hz
22 Hz
20 Hz
19 Hz
22 Hz
22 Hz
Et
22 Hz
(m)
iPr
c-Hex
Bn
Bn
Bn
H
47.4
19 Hz
(m)
20 Hz
46.6*
46.6*
46.7*
(m)
(m)
22 Hz
4g
4h
(m)
a
b
c
NMR solvent was D2O with TSP and products were measured as their HCl salts.
13C NMR chemical shifts for R, R1 are presented in Section 4.
Shifts marked with * are for CD2 carbons, which were observed as quintets except 4f, C-3 which was under the other shifts, and overlaid 4g shifts. See also Figure 2.
known compounds were re-evaluated, since the 13C NMR chemical
Table 7
13C NMR chemical shifts for intermediates 6a–f a,b,c
shifts differences in the literature compared to our shifts were
1–3 ppm (literature shifts measured in D2O without internal
standard) or in some cases chemical shifts were missing.
The isotopic distribution was estimated for all the deuterated
target compounds. The isotopic purity was 87% for 4g containing
eight deuterium atoms, 91–97% for compounds with four deute-
rium atoms and 94–99% for compounds with two deuterium atoms,
as shown in Tables 1–3. In addition, according to the 1H NMR and
ESI mass spectra, the overall deuterium stage of all the labelled
compounds (detected deuterium amount in the molecule versus
expected) was over 97%.
IR was used to study the C–D stretches in deuterated com-
pounds. According to literature the range for C–D stretches is 2200–
2080 cmꢀ1, which is beyond ‘normal’ IR vibrations.34 However, only
compound 1l clearly showed bands around this region
(w2230 cmꢀ1). In the other spectra, there were no such clear vi-
brations in this region, or the bands are so weak or they may remain
under the highly structured ammonium (NHþ3 , NH2þ) vibrations
(3000–2000 cmꢀ1).
Compd
C1
CN
R
N
H
C2
R
C-1
C-2
1JCD
CN
6a
6e
6d
6f
6b
6c
Et
Et
Bn
44.9
44.7
44.3
44.2
42.6
42.2
18.7
18.3*
18.8
18.4*
19.1
118.8
118.8
118.7
118.7
118.8
118.9
21 Hz
21 Hz
Bn
iPr
c-Hex
19.2
a
b
c
NMR solvent was CDCl3 with TMS.
13C NMR chemical shifts for R are presented in Section 4.
Shifts marked with * are for CD2 carbons, which were observed as quintets.
Table 8
13C NMR chemical shifts for intermediate DAP derivatives 11a–ha,b,c
C3
R
C1
Compd
N
C2 NH
Ns
C-2
Ns
R
C-1
C-3
11a
11b
11c
11d
11e
11f
Et
Et
Et
Bn
Bn
Bn
iPr
c-Hex
44.2
44.1
44.0
45.1
45.1
45.0
41.1
41.2
28.9
28.7
40.6
40.1*
40.0*
40.5
40.0*
39.8*
40.4
41.1
3. Conclusions
(m)
(m)
28.0*
28.5
28.4
27.6*
32.0
32.1
(m)
(m)
A general method was developed to synthesize several deute-
rium labelled N-alkylated diaminopropanes containing two or four
deuteriums in the non-exchangeable positions. These key in-
termediates were used to prepare mono-, di- and unsubstituted,
sterically hindered, as well as symmetrically and non-symmetri-
cally substituted polyamine derivatives, and also their symmetri-
cally or non-symmetrically deuterium labelled analogues, having
two to eight deuterium atoms in the PA backbone. According to the
ESI-MS and 1H NMR data, the isotopic purities were high (87–99%)
for all the labelled compounds. Benzyl alkylated derivatives were
found to be convenient precursors in the synthesis of a variety of
labelled, non-N-alkylated diaminopropanes, spermidines and
spermines, and they served also as an alternative route to mono-
substituted polyamines.
(m)
(m)
11g
11h
a
NMR solvent was CDCl3 with TMS.
13C NMR chemical shifts for R and for protecting groups (Ns) are presented in
b
Section 4.
c
Shifts marked with * are for CD2 carbons, which were observed as multiplets due
to dynamic effects at room temperature.
shown in Tables 4–7. However, 13C NMR spectra of the protected
intermediates were more complex due to dynamic effects caused
by the neighbouring protecting groups at room temperature and
only multiplets were observed for the CD2 groups (Tables 8–10). A
typical example of the isotope effects generated by deuterium
atoms is shown in Figure 2. Negative (upfield to the corresponding
CH2) deuterium induced isotope shifts were observed over one, two
and three bonds, as can be estimated also from Tables 4–10. The
isotopic shifts are additive, since deuterated C-3 resonance is
shifted more upfield when also C-2 is deuterated (see Tables
4,5,8,9). In some cases, 1H and 13C NMR chemical shifts for the
4. Experimental
4.1. Reagents
Dimethylformamide (DMF) was distilled and stored over 4 Å
molecular sieves, triethylamine (TEA) was dried over KOH and

552
M.R. Ha¨kkinen et al. / Tetrahedron 65 (2009) 547–562
Table 9
13C NMR chemical shifts for intermediate SPD derivatives 12a–g and alkylating agents 15a–d and 10a–ba,b,c
C7
C1
C3 C5
N
Ns
Y
Compd
R
N
X
C2
C6
C8
R
X
Y
C-1
C-2
C-3
C-5
C-6
C-7
C-8
12a
12b
12c
12d
12e
12f
12g
15a
15b
15c
15d
10a
10b
Et
Et
Et
Bn
Bn
iPr
c-Hex
Et
Et
Et
Et
H
Ns
Ns
Ns
Ns
Ns
Ns
Ns
Ns
Ns
Ns
Ns
Boc
Boc
NPht
NPht
NPht
NPht
NPht
NPht
NPht
Cl
Cl
I
I
Cl
44.3
44.3
44.1
45.1
45.1
40.5
41.4
44.4
44.3
44.4
44.3
37.4
37.4
27.5
27.3
26.6*
26.9
26.7
30.3
30.4
27.6
26.7*
27.6
26.7*
28.6
28.6
45.2
44.6*
44.5*
44.9
44.3*
45.1
47.3
47.2
47.2
46.8
46.7
46.8
46.8
47.2
47.1
46.7
46.7
46.8
46.4
25.4
25.4
25.4
25.2
25.2
25.2
25.2
25.4
25.4
29.0
28.9
25.4
28.9
25.7
25.7
25.7
25.6
25.6
25.7
25.7
29.3
29.3
30.1
30.1
29.3
30.1
37.1
37.1
37.1
37.1
37.1
37.2
37.2
44.4
44.4
5.9
(m)
(m)
(m)
(m)
45.1
45.1
(m)
(m)
44.4*
45.1
44.3*
45.0
45.0
(m)
(m)
6.0
44.3
5.8
H
I
a
b
c
NMR solvent was CDCl3 with TMS.
13C NMR chemical shifts for R and for protecting groups (Ns, Pht and Boc) are presented in Section 4.
Shifts marked with * are for CD2 carbons, which were observed as multiplets due to dynamic effects at room temperature.
Table 10
13C NMR chemical shifts for intermediate SPM derivatives 13a–f and 14a–ga,b,c
Ns
N
X
N
Compd
C11
C10
C7
R
C1
C3 C5
N
Ns
C12
N
Ns
C2
C6 C8
R1
R
R1
X
C-1
C-2
C-3
C-5
C-6
C-7
C-8
C-10
C-11
C-12
13a
13b
13c
13d
13e
13f
Et
Et
Bn
H
H
H
H
H
H
Et
Et
iPr
c-Hex
Et
Et
Et
Boc
Boc
Boc
Boc
Boc
Boc
Ns
Ns
Ns
Ns
Ns
44.4
44.4
45.2
45.1
40.5
41.4
44.4
44.4
40.2
41.4
45.2
45.1
45.1
27.5
27.3
26.9
26.0*
30.2
30.4
27.5
27.2
29.8
30.3
26.9
26.0*
26.0*
44.9
44.4*
44.7
44.0*
44.9
44.8
45.0
44.5*
44.4
44.8
44.8
44.1*
44.3*
47.1
47.0
46.7
46.6
46.6
46.6
47.2
47.0
46.5
46.6
46.8
46.7
46.7
25.0
25.0
24.8
24.8
24.8
24.7
25.0
24.9
24.5
24.7
24.8
24.8
24.8
25.1
25.1
25.0
25.0
25.1
25.1
25.0
24.9
24.5
24.7
25.0
25.0
24.9
47.0
47.0
47.0
47.0
46.9
46.9
47.2
47.0
46.5
46.6
47.1
47.1
47.1
45.2
45.2
45.2
45.2
45.2
45.1
45.0
44.5*
44.4
44.8
45.0
45.0
44.3*
28.6
28.6
28.6
28.6
28.6
28.6
27.5
27.2
29.8
30.3
27.5
27.5
26.6*
37.4
37.4
37.4
37.4
37.4
37.4
44.4
44.4
40.2
41.4
44.4
44.4
44.3
(m)
(m)
Bn
(m)
iPr
c-Hex
Et
14a**
14b
*
Et
(m)
(m)
14c**
14d**
14e
14f
14g
iPr
c-Hex
Bn
Bn
Ns
Ns
(m)
(m)
(m)
(m)
Bn
(m)
(m)
a
NMR solvent was CDCl3 with TMS, except for **, which were measured in CDCl3þDMSO-d6 with TMS.
b
c
13C NMR chemical shifts for R, R1 and for protecting groups (Ns and Boc) are presented in Section 4.
Shifts marked with * are for CD2 carbons, which were observed as multiplets due to dynamic effects at room temperature.
distilled, diethylether was dried over sodium-benzophenone ketyl
and distilled, dichloromethane (DCM) and chloroform were dried
over P2O5 and distilled. All other reagents were used without fur-
ther purification. Reactions were followed with TLC using analytical
4.2. Syntheses of polyamines
4.2.1. 3-Ethylamino-propionitrile 6a28,35
Prepared by the known method28 from 70% ethylamine (20.0 g,
311 mmol) and acrylonitrile (10.6 g, 200 mmol) to give 6a (17.93 g,
91%) as a colourless liquid, bp 75 ꢁC/10 mbar. IR (neat): 3313 (br),
pre-coated silica gel 60 F254 plates, while silica gel (63–200
Merck) was used for column chromatographic separations.
mm,
1H and 13C NMR spectra were measured on a Bruker Avance
(Bruker, Rheinstetter, Germany) 500 DRX spectrometer operating
at 500.13 and 125.76 MHz, respectively. TMS was used as an in-
ternal standard in organic solvents, and sodium 3-(trimethylsilyl)-
1-propionic acid (TSP) in D2O. Relaxation delay (D1) was set to 60 s
for 1H measurements and to 10 s for carbon experiments. 1H and
13C NMR chemical shifts were assigned based on standard 2D
techniques (1H–1H COSY, 1H–13C HSQC and HMBC).
2969–2834, 2247, 1455, 1375, 1130 cmꢀ1
;
1H NMR (CDCl3):
), 2.53 (2H, t,
J¼6.6 Hz, H-2), 1.68 (1H, br, NH), 1.13 (3H, t, J¼7.1 Hz, H-
); 13C NMR
(CDCl3): 43.5 (C- ), 15.2 (C- ), rest in Table 7; HRMS (ESI-MS):
d 2.94
(2H, t, J¼6.6 Hz, H-1), 2.69 (2H, q, J¼7.1 Hz, H-
a
b
d
a
b
calcd for (MþH) C5H11N2 99.0922, found 99.0922.
4.2.2. 3-Isopropylamino-propionitrile 6b36,37
Isopropylamine (5.00 g, 85 mmol) was added dropwise with
stirring to a solution of acrylonitrile (6.73 g, 127 mmol) in methanol
(10 mL) within 20 min. The stirring was continued for 30 min at
room temperature and for 1 h under reflux. Solvents were evapo-
rated in vacuo and the residue was distilled (bp 100 ꢁC/35 mbar) to
give 6b (7.48 g, 79%) as a colourless liquid. IR (neat): 3313 (br),
Isotope distribution measurements and high-resolution mass
spectra were conducted on a QSTAR XL hybrid quadrupole TOF in-
strument (Applied Biosystems, Foster City, CA) using the positive
electrospray ionization mode. Isopropylamine, amino acids and
peptides were usedastheinternal standardsinHRMSmeasurements.
IR spectra as KBr pelleting were recorded on Nexus 470 FT-IR.
Melting points were measured in open capillary tubes and are
uncorrected.
2966–2870, 2247, 1472, 1381, 1175 cmꢀ1
(2H, t, J¼6.7 Hz, H-1), 2.85 (1H, sep., J¼6.2 Hz, H-
2), 1.09 (1H, br, NH), 1.07 (6H, d, J¼6.2 Hz, H-
); 13C NMR (CDCl3):
;
1H NMR (CDCl3):
d 2.92
a
), 2.51 (2H, t, H-
b

M.R. Ha¨kkinen et al. / Tetrahedron 65 (2009) 547–562
553
Figure 1. Deuterium induced changes in 1H and 13C NMR chemical shifts during the deuteration of 6d to 6f. Top: 1H NMR; the exchanged proton signals disappear and the re-
maining coupling pattern is simplifying. Bottom: 13C NMR; the carbon signal is splitting into a multiplet due to 13C–2H J coupling (1JCDz21 Hz) depending on how many deuteriums
are attached to the carbon. Singlet for CH2, 1:1:1 triplet for CHD and 1:2:3:2:1 quintet for CD2. A: unlabelled (6d), B: labelling in progress, C: labelled (6f).
d
48.1 (C-
a
), 22.9 (2C-
b
), rest in Table 7; HRMS (ESI-MS): calcd for
liquid, bp 125 ꢁC/2 mbar. IR (neat): cmꢀ1 as reported earlier;36 1
H
(MþH) C6H13N2 113.1079, found 113.1074.
NMR (CDCl3): d 7.36–7.22 (5H, m, Ph), 3.81 (2H, s, H-a), 2.90 (2H, t,
J¼6.6 Hz, H-1), 2.48 (2H, t, J¼6.6 Hz, H-2), 1.54 (1H, br, NH); 13C
4.2.3. 3-Cyclohexylamino-propionitrile 6c35,38,39
Prepared as 6b from cyclohexylamine (7.40 g, 75 mmol) and
acrylonitrile (5.94 g, 112 mmol) to give 6c (9.76 g, 86%) as a colour-
less liquid, bp 140 ꢁC/12 mbar. IR (neat): 3422 (br), 2927, 2853,
NMR (CDCl3):
53.2 (C-
C10H13N2 161.1079, found 161.1082.
d 139.5 (Ph), 128.6 (2C, Ph), 128.1 (2C, Ph), 127.3 (Ph),
), rest in Table 7; HRMS (ESI-MS): calcd for (MþH)
a
2247, 1450, 1130 cmꢀ1; 1H NMR (CDCl3):
d
2.95 (2H, t, J¼6.7 Hz, H-
),1.90–1.83 (2H, m, H-beq),1.77–1.70 (2H,
m, H-geq), 1.65–1.58 (1H, m, H-deq), 1.31–1.01 (6H, m, H-gax dax bax
NH); 13C NMR (CDCl3):
56.0 (C- ), 33.6 (2C- ), 26.0 (C- ), 24.9 (2C-
), rest in Table 7; HRMS (ESI-MS): calcd for (MþH) C9H17N2
153.1392, found 153.1391.
4.2.5. 3-Ethylamino-2,2-2H2-propionitrile 6e
1), 2.53–2.43 (3H, m, H-2,
a
Nitrile 6a (4.0 g, 40.75 mmol) in the mixture of 0.1 M NaOD in
D2O (26 mL) and EtOD (26 mL) was stirred at room temperature for
24 h. The reaction mixture was saturated with NaCl and extracted
with dry Et2O. Combined organic extracts were dried over K2CO3,
solvents were removed by distillation, after the product was
distilled under reduced pressure (2.65 g, 65%, 92% d, bp 72 ꢁC/
10 mbar). The reaction cycle was repeated once to give 6e (1.91 g,
47%, 98% d) as a colourless liquid. IR (neat): 3312 (br), 2969–2340,
,
,
,
d
a
b
d
g
4.2.4. 3-Benzylamino-propionitrile 6d36,40
Prepared as 6b from benzylamine (8.57 g, 80 mmol) and acry-
lonitrile (6.37 g, 120 mmol) to give 6d (10.75 g, 84%) as a colourless
2248, 1453, 1384, 1126 cmꢀ1 1H NMR (CDCl3):
; d 2.92 (2H, s, H-1),

554
M.R. Ha¨kkinen et al. / Tetrahedron 65 (2009) 547–562
Figure 2. Negative deuterium induced isotope shifts, Dd, in 13C NMR spectra were observed over one (0.7 ppm), two (0.17 ppm) and three bonds (0.06 ppm), as demonstrated for
compounds 4e, 4f and 4g.
2.69 (2H, q, J¼7.1 Hz, H-
a
), 1.13 (3H, t, J¼7.1 Hz, H-
b
); 13C NMR
catalyst was filtered off through Celite, washed with hot ethanol
and water, and the combined filtrates were evaporated to dryness
in vacuo. The residue was recrystallized from ethanol–EtOAc to
give 1.
(CDCl3): 43.4 (C- ), 15.2 (C-b
d
a
), rest in Table 7; HRMS (ESI-MS):
calcd for (MþH) C5H9D2N2 101.1048, found 101.1044.
4.2.6. 3-Benzylamino-2,2-2H2-propionitrile 6f
Method C. Following the published procedure for N1-benzyl-
butane-1,4-diamine,41 compound 1 was prepared from the corre-
sponding aldehyde (20.5 mmol) and (3-amino-propyl)carbamic
acid tert-butyl ester (7, 22.6 mmol) after reduction and amino
group deprotection.
Prepared as 6e from 6d (3.2 g, 20 mmol) to give 6f (2.62 g, 81%,
95% d, bp 120 ꢁC/0.9 mbar) after the first cycle. The reaction cycle
was repeated once to give 6f (2.16 g, 66%, 99% d) as a colourless
liquid. IR (neat): 3336 (br), 3085–2843, 2248, 1454, 743, 700 cmꢀ1
;
1H NMR (CDCl3):
s, H-1); 13C NMR (CDCl3):
127.3 (Ph), 53.2 (C- ), rest in Table 7; HRMS (ESI-MS): calcd for
d
7.36–7.22 (5H, m, Ph), 3.82 (2H, s, H-
a), 2.91 (2H,
d
139.5 (Ph), 128.6 (2C, Ph), 128.1 (2C, Ph),
4.3.1. N1-Ethylpropane-1,3-diamine dihydrochloride 1a39
a
Prepared from 6a to give 1a (method A: 43%, method B: 80%) as
colourless solid, mp 224–225 ꢁC. IR (KBr): 3000–2387, 1606, 1459,
(MþH) C10H11D2N2 163.1204, found 163.1209.
1379, 804 cmꢀ1; 1H NMR (D2O):
d
3.17–3.10 (6H, m, H-
2.06 (2H, m, H-2), 1.30 (3H, t, J¼7.3 Hz, H-
); 13C NMR (D2O):
(C- ), 13.3 (C-
a
,1,3), 2.12–
4.3. General protocols to prepare 1a–k
b
d
45.9
a
b), rest in Table 4; HRMS (ESI-MS): calcd for (MþH)
Method A. To a cooled (0 ꢁC) suspension of LiAlH4 or LiAlD4
(20.4 mmol) in dry Et2O (40 mL), a solution of nitrile 6 (500 mg,
10 mmol) in dry Et2O (10 mL) was added dropwise with stirring
over a period of 45 min. The reaction mixture was allowed to warm
to room temperature, heated at reflux for 3 h and further stirred at
room temperature for 20 h. The reaction was quenched at 0–4 ꢁC by
dropwise addition of water (1.2 mL) and NaOH (5 M, 7.1 mL) with
stirring. The organic phase was separated and the residue was
extracted with hot chloroform (4ꢂ20 mL). The combined organic
extracts were dried over MgSO4 and the crude product was pre-
cipitated by treating with dry HCl. The subsequent recrystallization
from ethanol–EtOAc yielded 1.
C5H15N2 103.1235, found 103.1229.
4.3.2. N1-Ethyl-3,3-2H2-propane-1,3-diamine dihydrochloride 1b
Prepared from 6a to give 1b (method A, 47%, 99% d) as a col-
ourless solid, mp 219–221 ꢁC. IR (KBr): 3000–2386, 1604, 1460,
1382, 1144, 855, 804 cmꢀ1
a
;
1H NMR (D2O):
d
3.17–3.10 (4H, m, H-
,1), 2.11–2.05 (2H, m, H-2), 1.30 (3H, t, J¼7.3 Hz, H-
); 13C NMR
(D2O): 45.9 (C- ), 13.4 (C- ), rest in Table 4; HRMS (ESI-MS): calcd
b
d
a
b
for (MþH) C5H13N2D2 105.1361, found 105.1354. Isotope distribu-
tion: 0% d0, 1% d1, 98% d2.
4.3.3. N1-Ethyl-2,2-2H2-propane-1,3-diamine dihydrochloride 1c
Prepared from 6e to give 1c (method A, 43%, 97% d) as a col-
ourless solid, mp 222–224 ꢁC. IR (KBr): 3000–2369, 1594, 1462,
Method B. To a solution of 6 (10.2 mmol) in ethanol (30 mL) was
added concd HCl (2 mL) and the mixture was deoxygenated with
argon for 10 min. Adam’s catalyst (PtO2, 0.44 mmol) was added and
the mixture was hydrogenated at 3.5 bar pressure for 20 h. The
1374, 1185, 802 cmꢀ1
1.30 (3H, t, J¼7.3 Hz, H-
;
1H NMR (D2O):
); 13C NMR (D2O):
d
3.17–3.08 (6H, m, H-
a
,1,3),
),
b
d
45.9 (C- ), 13.4 (C-
a
b

M.R. Ha¨kkinen et al. / Tetrahedron 65 (2009) 547–562
555
rest in Table 4; HRMS (ESI-MS): calcd for (MþH) C5H13N2D2
1.25–1.13 (1H, m, H-dax); 13C NMR (D2O):
27.3 (C- ), 26.8 (2C- ), rest in Table 4; HRMS (ESI-MS): calcd for
d 60.3 (C-a), 31.7 (2C-b),
105.1361, found 105.1356. Isotope distribution: 0% d0, 6% d1, 94% d2.
d
g
(MþH) C9H21N2 157.1705, found 157.1707.
4.3.4. N1-Ethyl-2,2,3,3-2H4-propane-1,3-diamine dihydrochloride 1d
Prepared from 6e to give 1d (method A, 47%, 98% d) as a colour-
less solid, mp 218–220 ꢁC. IR (KBr): 3000–2388, 1603, 1463, 1382,
4.3.11. N1-Cyclohexyl-3,3-2H2-propane-1,3-diamine dihydrochloride 1k
Prepared from 6c to give 1k (method A, 70%, 99% d) as
a colourless solid, mp 205–207 ꢁC. IR (KBr): 3000–2385, 1604,
1171, 837, 806 cmꢀ1; 1H NMR (D2O):
); 13C NMR (D2O):
d
3.17–3.10 (4H, m, H-
a
,1), 1.30
(3H, t, J¼7.3 Hz, H-
b
d
46.0 (C- ), 13.4 (C-
a
b
), rest in
1467, 855 cmꢀ1
;
1H NMR (D2O):
d
3.22–3.08 (3H, m, H-1,
2.13–2.02 (4H, m, H-2,beq), 1.90–1.79 (2H, m, H-geq), 1.72–1.63
(1H, m, H-deq), 1.41–1.27 (4H, m, H-bax gax), 1.25–1.13 (1H, m,
H-dax); 13C NMR (D2O):
60.3 (C- ), 31.7 (2C- ), 27.3 (C- ), 26.7
(2C-
a),
Table 4; HRMS (ESI-MS): calcd for (MþH) C5H11N2D4 107.1486,
found 107.1480. Isotope distribution: 0% d0–d2, 8% d3, 92% d4.
,
d
a
b
d
4.3.5. N1-Benzyl-propane-1,3-diamine dihydrochloride 1e42,43
Prepared from 6d to give 1e (method A, 74%) as a colourless
solid, mp 271–272 ꢁC.
g
), rest in Table 4; HRMS (ESI-MS): calcd for (MþH)
C9H19N2D2 159.1830, found 159.1826. Isotope distribution: 0%
d0, 2% d1, 98% d2.
Prepared from benzaldehyde (2.2 g, 20.5 mmol) and (3-amino-
propyl)carbamic acid tert-butyl ester (3.9 g, 22.6 mmol) following
method C, which after recrystallization from ethanol–EtOAc gave
1e in the overall (3 steps) with a 56% yield. IR (KBr): 3050–2410,
4.3.12. 1,1-2H2-Propane-1,3-diamine dihydrochloride 1l46
To a mixture of 1f (190 mg, 0.79 mmol) in ethanol (30 mL),
water was added to dissolve 1f followed with 10% Pd–C (80 mg)
and the reaction mixture was hydrogenated at 3.5 bar pressure for
20 h. The catalyst was filtered off through Celite, washed with
ethanol and water, and the combined filtrates were evaporated to
dryness in vacuo. Recrystallization from ethanol–water–EtOAc
yielded 1l (92 mg, 78%, 99% d) as a colourless solid, mp 249–250 ꢁC
(lit. mp 246–250 ꢁC).46 IR (KBr): 3000–2300, 2227, 1610, 1463, 1167,
1599, 1484, 1174, 744, 697 cmꢀ1; 1H NMR (D2O):
Ph), 4.29 (2H, s, H-
3), 2.17–2.07 (2H, m, H-2); 13C NMR (D2O):
Ph), 132.6 (Ph), 132.2 (2C, Ph), 54.1 (C- ), rest in Table 4; HRMS (ESI-
d
7.57–7.48 (5H, m,
), 3.24–3.17 (2H, m, H-1), 3.14–3.08 (2H, m, H-
133.3 (Ph), 132.7 (2C,
a
d
a
MS): calcd for (MþH) C10H17N2 165.1392, found 165.1389.
4.3.6. N1-Benzyl-3,3-2H2-propane-1,3-diamine dihydrochloride 1f
Prepared from 6d to give 1f (method A, 83%, 99% d) as a col-
ourless solid, mp 275 ꢁC. IR (KBr): 3050–2387, 1601, 1478, 854, 744,
844 cmꢀ1; 1H NMR (D2O):
d
3.15–3.09 (2H, m, H-1), 2.10–2.04 (2H,
in Table 4; HRMS (ESI-MS): calcd for
m, H-2); 13C NMR (D2O):
d
(MþH) C3H9N2D2 77.1048, found 77.1051. Isotope distribution: 1%
697 cmꢀ1; 1H NMR (D2O):
d
7.57–7.48 (5H, m, Ph), 4.29 (2H, s, H-
3.24–3.17 (2H, m, H-1), 2.16–2.07 (2H, m, H-2); 13C NMR (D2O):
133.5 (Ph), 132.7 (2C, Ph), 132.6 (Ph), 132.1 (2C, Ph), 54.1 (C- ), rest
a),
d0, 1% d1, 98% d2.
d
a
4.3.13. N1-Ethyl-bis-N1,N3-(2-nitrobenzenesulfonyl)propane-1,3-
in Table 4; HRMS (ESI-MS): calcd for (MþH) C10H15N2D2 167.1517,
diamine 11a
found 167.1513. Isotope distribution: 0% d0, 2% d1, 98% d2.
A
suspension of 1a (2.0 g, 11.42 mmol) and TEA (8 mL,
57 mmol) in dry DCM (100 mL) was cooled to 0 ꢁC. NsCl (5.32 g,
23.99 mmol) in dry DCM (70 mL) was added dropwise with stir-
ring over a period of 10 min, and stirring was continued for 1 h at
0 ꢁC and then for 3 h at room temperature. The reaction mixture
was washed with water (2ꢂ200 mL), 10% citric acid (1ꢂ200 mL),
brine (1ꢂ200 mL), dried over MgSO4 and evaporated to dryness in
vacuo that resulted in 11a (5.4 g, w100%) as a pale yellow solid,
mp 82–84 ꢁC. IR (KBr): 3321, 3095–2878, 1545, 1380, 1160,
4.3.7. N1-Benzyl-2,2,3,3-2H4-propane-1,3-diamine
dihydrochloride 1g
Prepared from 6f to give 1g (method A, 88%, 98% d) as a col-
ourless solid, mp 274–275 ꢁC. IR (KBr): 3050–2429, 1600, 1468, 836,
744, 697 cmꢀ1; 1H NMR (D2O):
d
7.57–7.46 (5H, m, Ph), 4.29 (2H, s,
), 3.19 (2H, s, H-1); 13C NMR (D2O):
133.3 (Ph), 132.7 (2C, Ph),
132.6 (Ph),132.2 (2C, Ph), 54.1 (C- ), rest in Table 4; HRMS (ESI-MS):
H-a
d
a
calcd for (MþH) C10H13N2D4 169.1643, found 169.1641. Isotope
736 cmꢀ1; 1H NMR (CDCl3):
m, Ph), 7.87–7.83 (1H, m, Ph), 7.78–7.67 (4H, m, Ph), 7.64–7.59 (1H,
m, Ph), 5.66 (1H, t, J¼6.2 Hz, NH), 3.42–3.31 (4H, m, H- ,1), 3.22–
3.16 (2H, m, H-3), 1.89–1.81 (2H, m, H-2), 1.10 (3H, t, J¼7.1 Hz, H- );
13C NMR (CDCl3):
148.1, 148.0, 133.7, 133.6 (2C), 133.3, 132.9,
131.8, 130.9, 130.8, 125.5, 124.2 (totally 12C–Ph), 42.4 (C- ), 13.7 (C-
), rest in Table 8; HRMS (ESI-MS): calcd for (MþH) C17H21N4O8S2
473.0801, found 473.0793.
d 8.14–8.09 (1H, m, Ph), 8.03–7.98 (1H,
distribution: 0% d0–d2, 7% d3, 92% d4.
a
4.3.8. N1-Isopropylpropane-1,3-diamine dihydrochloride 1h39,44
Prepared from 6b to give 1h (method A: 58%, method B: 83%) as
a colourless solid, mp 184–186 ꢁC. IR (KBr): 3000–2385, 1597, 1473,
b
d
a
1379, 1136, 766 cmꢀ1; 1H NMR (D2O):
d
3.45 (1H, sep., J¼6.6 Hz, H-
3.19–3.09 (4H, m, H-1,3), 2.13–2.03 (2H, m, H-2),1.33 (6H, d, J¼6.6 Hz,
H- 53.9 (C- ), 21.0 (2C- ), rest in Table 4; HRMS
); 13C NMR (D2O):
a
),
b
b
d
a
b
(ESI-MS): calcd for (MþH) C6H17N2 117.1392, found 117.1386.
4.3.14. N1-Ethyl-bis-N1,N3-(2-nitrobenzenesulfonyl)-3,3-2H2-
propane-1,3-diamine 11b
4.3.9. N1-Isopropyl-3,3-2H2-propane-1,3-diamine dihydrochloride 1i
Prepared from 6b to give 1i (method A, 59%, 99% d) as a col-
ourless solid, mp 182–184 ꢁC. IR (KBr): 3000–2300, 1594, 1471,
Prepared as 11a from 1b (1.20 g, 6.78 mmol) to give 11b (3.33 g,
w100%) as a pale yellow solid, mp 79–81 ꢁC. IR (KBr): 3323, 3095–
2879, 1545, 1380, 1161, 732 cmꢀ1; 1H NMR (CDCl3):
d 8.14–8.09 (1H,
1382, 1161, 845 cmꢀ1; 1H NMR (D2O):
a
d
3.45 (1H, sep., J¼6.6 Hz, H-
), 3.18–3.13 (2H, m, H-1), 2.10–2.04 (2H, m, H-2), 1.33 (6H, d,
); 13C NMR (D2O):
53.9 (C- ), 21.0 (2C- ), rest in
m, Ph), 8.02–7.97 (1H, m, Ph), 7.87–7.84 (1H, m, Ph), 7.78–7.67 (4H,
m, Ph), 7.64–7.59 (1H, m, Ph), 5.66 (1H, br, NH), 3.42–3.31 (4H, m,
J¼6.6 Hz, H-
b
d
a
b
H-
(CDCl3):
130.8, 125.5, 124.2 (totally 12C–Ph), 42.4 (C-
a
,1), 1.86–1.81 (2H, m, H-2), 1.10 (3H, t, J¼7.1 Hz, H-
148.1, 148.0, 133.7, 133.6, 133.6, 133.3, 132.9, 131.8, 130.9,
), 13.7 (C- ), rest in
b
); 13C NMR
Table 4; HRMS (ESI-MS): calcd for (MþH) C6H15N2D2 119.1517,
d
found 119.1514. Isotope distribution: 0% d0, 1% d1, 99% d2.
a
b
Table 8; HRMS (ESI-MS): calcd for (MþH) C17H19D2N4O8S2
4.3.10. N1-Cyclohexylpropane-1,3-diamine dihydrochloride 1j39,45
Prepared from 6c to give 1j (method A: 65%, method B: 84%) as
a colourless solid, mp 200–202 ꢁC (lit. mp 199–202 ꢁC).45 IR (KBr):
475.0926, found 475.0925.
4.3.15. N1-Ethyl-bis-N1,N3-(2-nitrobenzenesulfonyl)-2,2,3,3-2H4-
propane-1,3-diamine 11c
Prepared as 11a from 1d (700 mg, 3.91 mmol) to give 11c
(1.89 g, w100%) as a pale yellow solid, mp 79–83 ꢁC. IR (KBr): 3323,
3000–2368, 1606, 1464, 1164, 783 cmꢀ1
3.08 (5H, m, H-1,3,
H-geq), 1.72–1.63 (1H, m, H-deq), 1.41–1.27 (4H, m, H-bax
;
1H NMR (D2O):
d
3.22–
), 2.13–2.02 (4H, m, H-2,beq), 1.90–1.79 (2H, m,
gax),
a
,

556
M.R. Ha¨kkinen et al. / Tetrahedron 65 (2009) 547–562
3095–2881, 1544, 1380, 1161, 746 cmꢀ1
8.10 (1H, m, Ph), 8.04–7.99 (1H, m, Ph), 7.89–7.84 (1H, m, Ph), 7.78–
7.67 (4H, m, Ph), 7.64–7.60 (1H, m, Ph), 5.63 (1H, br, NH), 3.38 (2H, s,
;
1H NMR (CDCl3):
d
8.14–
4.3.20. N1-Cyclohexyl-bis-N1,N3-(2-nitrobenzenesulfonyl)propane-
1,3-diamine 11h
Prepared as 11g from 1j (917 mg, 4.0 mmol) to give 11h (1.02 g,
48%) as a pale yellow solid, mp 134–136 ꢁC. Rf 0.50 (EtOAc–hexane
3:2); IR (KBr): 3311, 3091–2851, 1542, 1164 cmꢀ1; 1H NMR (CDCl3):
H-1), 3.35 (2H, q, J¼7.2 Hz, H-
(CDCl3): 148.1, 148.0, 133.7, 133.6, 133.6, 133.2, 132.9 131.8, 130.9,
130.8, 125.5, 124.2 (totally 12C–Ph), 42.4 (C- ), 13.7 (C- ), rest in
a
), 1.11 (3H, t, J¼7.2 Hz, H-
b
); 13C NMR
d
a
b
d
8.15–8.11 (1H, m, Ph), 8.04–7.99 (1H, m, Ph), 7.89–7.85 (1H, m,
Ph), 7.78–7.66 (4H, m, Ph), 7.61–7.56 (1H, m, Ph), 5.62 (1H, t,
J¼6.3 Hz, NH), 3.70–3.62 (1H, m, H- ), 3.37–3.32 (2H, m, H-1),
3.24–3.18 (2H, m, H-3), 1.92–1.85 (2H, m, H-2), 1.78–1.72 (2H, m,
H-geq), 1.69–1.59 (3H, m, H-beq deq), 1.40–1.25 (4H, m, H-bax gax),
1.10–0.99 (1H, m, H-dax); 13C NMR (CDCl3):
148.1 (2C), 133.8,
133.7, 133.6, 133.5, 132.9, 131.7, 130.9, 130.8, 125.5, 124.1 (totally
12C–Ph), 58.3 (C- ), 32.1 (2C- ), 26.0 (2C- ), 25.2 (C- ), rest in
Table 8; HRMS (ESI-MS): calcd for (MþH) C17H17D4N4O8S2 477.1052,
found 477.1046.
a
4.3.16. N1-Benzyl-bis-N1,N3-(2-nitrobenzenesulfonyl)propane-1,3-
diamine 11d
,
,
d
Prepared as 11a from 1e (600 mg, 2.53 mmol) to give 11d
(1.25 g, 92%) as a pale yellow solid, mp 40–55 ꢁC (amorphous solid).
a
b
g
d
IR (KBr): 3342 (br), 3095–2880, 1541, 1163, 732 cmꢀ1
;
1H NMR
Table 8; HRMS (ESI-MS): calcd for (MþNa) C21H26N4O8S2Na
(CDCl3):
(1H, m, Ph), 7.74–7.64 (5H, m, Ph), 7.31–7.22 (5H, m, Ph), 5.39 (1H, t,
J¼6.3 Hz, NH), 4.47 (2H, s, H- ), 3.36–3.32 (2H, m, H-1), 3.02–2.96
(2H, m, H-3), 1.62–1.55 (2H, m, H-2); 13C NMR (CDCl3):
148.0,
148.0, 135.5, 133.8, 133.6, 133.6, 133.1, 132.8, 131.9, 131.1, 130.9, 128.9
(2C), 128.3 (2C), 128.3, 125.4, 124.3 (totally 18C–Ph), 52.3 (C- ), rest
d
8.07–8.03 (1H, m, Ph), 8.01–7.98 (1H, m, Ph), 7.86–7.82
549.1090, found 549.1098.
a
4.3.21. 2-(4-{[3-(Ethyl-(2-nitrobenzenesulfonyl)amino)propyl]-(2-
nitrobenzenesulfonyl)amino}-butyl)isoindole-1,3-dione 12a
d
To a mixture of 11a (1.0 g, 2.17 mmol) and K2CO3 (0.88 g,
6.35 mmol) in dry DMF (25 mL), N-(4-iodobutyl)phthalimide32
(0.77 g, 2.33 mmol) was added with stirring and the mixture was
allowed to react at room temperature for 17 h. After filtration, the
solvents were evaporated in vacuo and the residue was purified on
silica gel using first EtOAc–hexane 1:1 and then 5:2 as an eluent to
give 12a (1.24 g, 85%) as a colourless solid, mp 44–46 ꢁC (amor-
phous solid). Rf 0.53 (EtOAc–hexane 5:2); IR (KBr): 3095, 2940 (br),
a
in Table 8; HRMS (ESI-MS): calcd for (MþH) C22H23N4O8S2
535.0957, found 535.0984.
4.3.17. N1-Benzyl-bis-N1,N3-(2-nitrobenzenesulfonyl)-3,3-2H2-propane-
1,3-diamine 11e
Prepared as 11a from 1f (700 mg, 2.93 mmol) to give 11e
(1.43 g, 91%) as a pale yellow solid, mp 40–48 ꢁC (amorphous
1709, 1543, 1373, 1158, 720 cmꢀ1; 1H NMR (CDCl3):
m, Ph), 7.86–7.81 (2H, m, Ph), 7.74–7.55 (8H, m, Ph), 3.68–3.64 (2H,
m, H-8), 3.39–3.26 (8H, m, H- ,1,3,5), 1.91–1.83 (2H, m, H-2), 1.69–
1.62 (2H, m, H-7) 1.60–1.52 (2H, m, H-6), 1.12 (3H, t, J¼7.1 Hz, H- );
13C NMR (CDCl3):
168.4 (2C, C]O), 148.0 (2C), 134.0 (2C), 133.5,
133.5, 133.4, 133.2, 132.1 (2C), 131.8, 131.8, 130.8 (2C), 124.2 (2C),
123.3 (2C) (totally 18C–Ph), 42.3 (C- ), 13.6 (C- ), rest in Table 9;
d 8.02–7.98 (2H,
solid). IR (KBr): 3341 (br), 3095–2880, 1541, 1163, 740, 699 cmꢀ1
;
1H NMR (CDCl3):
7.86–7.82 (1H, m, Ph), 7.74–7.64 (5H, m, Ph), 7.31–7.22 (5H, m, Ph),
5.38 (1H, br, NH), 4.47 (2H, s, H- ), 3.36–3.32 (2H, m, H-1), 1.59–
1.55 (2H, m, H-2); 13C NMR (CDCl3):
148.0 (2C), 135.5, 133.8,
133.7 133.6, 133.1, 132.8, 131.9, 131.1, 130.9, 128.9 (2C), 128.3 (2C),
128.2, 125.4, 124.3 (totally 18C–Ph), 52.3 (C- ), rest in Table 8;
d
8.07–8.03 (1H, m, Ph), 8.01–7.98 (1H, m, Ph),
a
b
a
d
d
a
b
a
HRMS (ESI-MS): calcd for (MþH) C29H32N5O10S2 674.1591, found
HRMS (ESI-MS): calcd for (MþH) C22H21D2N4O8S2 537.1083, found
674.1584.
537.1083.
4.3.22. 2-(4-{[3-(Ethyl-(2-nitrobenzenesulfonyl)-amino)-1,1-2H2-
propyl]-(2-nitrobenzenesulfonyl)-amino}butyl) isoindole-1,3-
dione 12b
4.3.18. N1-Benzyl-bis-N1,N3-(2-nitrobenzenesulfonyl)-2,2,3,3-2H4-
propane-1,3-diamine 11f
Prepared as 11a from 1g (2.50 g, 10.36 mmol) to give 11f (5.16 g,
93%) as a pale yellow solid, mp 39–48 ꢁC (amorphous solid). IR
Prepared as 12a from 11b (600 mg, 1.26 mmol) to give 12b
(803 mg, 94%) as a colourless solid, mp 38–44 ꢁC (amorphous
solid). Rf 0.53 (EtOAc–hexane 5:2); IR (KBr): 3095, 2940 (br), 1710,
(KBr): 3340 (br), 3084–2928, 2225, 1541, 1164, 740 cmꢀ1 1H NMR
;
(CDCl3):
(1H, m, Ph), 7.75–7.63 (5H, m, Ph), 7.31–7.21 (5H, m, Ph), 5.41 (1H,
br, NH), 4.46 (2H, s, H- 148.0
), 3.32 (2H, s, H-1); 13C NMR (CDCl3):
(2C), 135.5, 133.8, 133.7, 133.5, 133.0, 132.9, 132.0, 130.9, 130.8, 128.8
(2C), 128.3 (2C), 128.2, 125.4, 124.3 (totally 18C–Ph), 52.2 (C- ), rest
d
8.06–8.01 (1H, m, Ph), 7.99–7.95 (1H, m, Ph), 7.84–7.80
1544, 1373, 1160, 721 cmꢀ1
;
1H NMR (CDCl3):
d
8.02–7.97 (2H, m,
Ph), 7.86–7.81 (2H, m, Ph), 7.75–7.55 (8H, m, Ph), 3.68–3.63 (2H,
m, H-8), 3.39–3.27 (6H, m, H- ,1,5), 1.88–1.83 (2H, m, H-2), 1.69–
1.62 (2H, m, H-7), 1.60–1.52 (2H, m, H-6), 1.11 (3H, t, J¼7.1 Hz,
H- 168.4 (2C, C]O), 148.0 (2C), 134.0 (2C),
); 13C NMR (CDCl3):
133.6, 133.5, 133.3, 133.1, 132.0 (2C), 131.8, 131.8, 130.7 (2C), 124.2
(2C), 123.3 (2C) (totally 18C–Ph), 42.3 (C- ), 13.6 (C- ), rest in
a
d
a
a
b
d
in Table 8; HRMS (ESI-MS): calcd for (MþH) C22H19D4N4O8S2
539.1208, found 539.1208.
a
b
Table 9; HRMS (ESI-MS): calcd for (MþH) C29H30D2N5O10S2
4.3.19. N1-Isopropyl-bis-N1,N3-(2-nitrobenzenesulfonyl)propane-
1,3-diamine 11g
676.1716, found 676.1721.
Prepared as 11a from 1h (3.00 g, 15.86 mmol) but after 2 h at
room temperature, the reaction mixture was heated at reflux for
20 h to give 11g (3.08 g, 40%) as a pale yellow solid after column
chromatography using EtOAc–hexane 3:2 as an eluent, mp 100–
102 ꢁC. Rf 0.52 (EtOAc–hexane 3:2); IR (KBr): 3332, 3096–2850,
4.3.23. 2-(4-{[3-(Ethyl-(2-nitrobenzenesulfonyl)-amino)-
1,1,2,2-2H4-propyl]-(2-nitrobenzenesulfonyl)-
amino}butyl)isoindole-1,3-dione 12c
Prepared as 12a from 11c (715 mg, 1.50 mmol) to give 12c
(948 mg, 93%) as a colourless solid, mp 41–45 ꢁC (amorphous solid).
Rf 0.57 (EtOAc–hexane 5:2); IR (KBr): 3095, 2937 (br), 1710, 1544,
1545, 1380, 1160 cmꢀ1 1H NMR (CDCl3):
; d 8.15–8.12 (1H, m, Ph),
8.05–8.01 (1H, m, Ph), 7.89–7.85 (1H, m, Ph), 7.78–7.66 (4H, m,
Ph), 7.62–7.58 (1H, m, Ph), 5.61 (1H, br, NH), 4.10 (1H, sep.,
1372, 1161, 721 cmꢀ1
;
1H NMR (CDCl3):
d
8.04–7.96 (2H, m, Ph),
7.86–7.81 (2H, m, Ph), 7.75–7.54 (8H, m, Ph), 3.68–3.63 (2H, m, H-8),
3.39–3.27 (6H, m, H- ,1,5), 1.69–1.61 (2H, m, H-7), 1.60–1.52 (2H, m,
H-6),1.12 (3H, t, J¼7.1 Hz, H- 168.4 (2C, C]O),
); 13C NMR (CDCl3):
148.0 (2C), 134.0 (2C), 133.6, 133.5, 133.3, 133.1, 132.0 (2C), 131.8,
131.8, 130.7 (2C), 124.2 (2C), 123.3 (2C) (totally 18C–Ph), 42.3 (C- ),
13.6 (C-
), rest in Table 9; HRMS (ESI-MS): calcd for (MþH)
C29H28D4N5O10S2 678.1842, found 678.1848.
J¼6.8 Hz, H-
1.94–1.87 (2H, m, H-2), 1.14 (6H, d, J¼6.8 Hz, H-
(CDCl3): 148.1 (2C), 133.7, 133.6, 133.6, 133.5, 132.9, 131.7, 131.0,
131.0, 125.5, 124.2 (totally 12C–Ph), 50.1 (C- ), 21.4 (2C- ), rest in
a
), 3.36–331 (2H, m, H-1), 3.23–3.19 (2H, m, H-3),
a
b
); 13C NMR
b
d
d
a
b
a
Table 8; HRMS (ESI-MS): calcd for (MþH) C18H23N4O8S2 487.0957,
b
found 487.0957.

M.R. Ha¨kkinen et al. / Tetrahedron 65 (2009) 547–562
557
4.3.24. 2-(4-{[3-(Benzyl-(2-nitrobenzenesulfonyl)amino)propyl]-
4.3.28. N1-(3-Ethylamino-propyl)-butane-1,4-diamine
trihydrochloride 2a47
(2-nitrobenzenesulfonyl)amino}-butyl)isoindole-1,3-dione 12d
Prepared as 12a from 11d (1.19 g, 2.23 mmol) to give 12d (1.32 g,
81%) as a colourless solid, mp 47–50 ꢁC (amorphous solid). Rf 0.57
(EtOAc–hexane 5:2); IR (KBr): 3093, 2940, 1710, 1543, 1160,
A mixture of 12a (1.21 g, 1.79 mmol), PhSH (0.56 mL, 5.40 mmol)
and K2CO3 (1.48 g, 10.74 mmol) in DMF (15 mL) was stirred over-
night at room temperature. The reaction mixture was then evap-
orated to dryness in vacuo. The residue was dissolved in a mixture
of DCM and 2 M KOH (2:1, 150 mL), the water layer was extracted
with DCM (3ꢂ50 mL), and the combined organic extracts were
dried over MgSO4. Solvent was evaporated in vacuo and the residue
721 cmꢀ1; 1H NMR (CDCl3):
d
8.01–7.98 (1H, m, Ph), 7.95–7.91 (1H,
m, Ph), 7.85–7.81 (2H, m, Ph), 7.74–7.60 (7H, m, Ph), 7.56–7.52 (1H,
m, Ph), 7.34–7.25 (5H, m, Ph), 4.48 (2H, s, H- ), 3.63–3.58 (2H, m, H-
a
8), 3.23–3.18 (2H, m, H-1), 3.16–3.08 (4H, m, H-3,5), 1.68–1.61 (2H,
m, H-2), 1.60–1.52 (2H, m, H-7), 1.45–1.36 (2H, m, H-6); 13C NMR
was dissolved in ethanol (14 mL) containing N2H4$H2O (135 mL,
(CDCl3):
133.5, 133.2 (2C), 132.1 (2C), 131.9, 131.7, 131.0, 130.7, 128.8 (2C),
128.4 (2C),128.2,124.3,124.1,123.3 (2C) (totally 24C–Ph), 51.9 (C- ),
d
168.3 (2C, C]O), 148.0, 147.9, 135.6, 134.0 (2C), 133.7,
2.78 mmol). After heating for 3 h at reflux, the solvents were
evaporated in vacuo, the residue was suspended in a mixture of
dioxane and concd HCl (3:1, 28 mL) followed by evaporating to
dryness in vacuo. After being co-evaporated once with dioxane,
DCM was added to the residue and the product was filtered,
washed with DCM and extracted with water. The filtrate was
evaporated in vacuo, dissolved in water, the insoluble phthalyl
hydrazide was filtered off and the solvent was evaporated to dry-
ness in vacuo. The treatment with water was repeated twice. The
product was washed with EtOAc, DCM and cold ethanol. After re-
crystallization from ethanol–water–EtOAc, 2a (376 mg, 74%) was
obtained as a colourless solid, mp >298 ꢁC decomp. IR (KBr): 3000–
a
rest in Table 9; HRMS (ESI-MS): calcd for (MþH) C34H34N5O10S2
736.1747, found 736.1746.
4.3.25. 2-(4-{[3-(Benzyl-(2-nitrobenzenesulfonyl)amino)-1,1-2H2-
propyl]-(2-nitrobenzenesulfonyl)-amino}butyl)isoindole-1,3-
dione 12e
Prepared as 12a from 11e (1.42 g, 2.61 mmol) to give 12e (1.83 g,
95%) as colourless solid, mp 50–52 ꢁC (amorphous solid). Rf 0.47
(EtOAc–hexane 3:2); IR (KBr): 3093–2980, 1709, 1541, 1161,
721 cmꢀ1; 1H NMR (CDCl3):
d
8.01–7.98 (1H, m, Ph), 7.95–7.91 (1H,
m, Ph), 7.85–7.81 (2H, m, Ph), 7.74–7.60 (7H, m, Ph), 7.56–7.52 (1H,
m, Ph), 7.33–7.25 (5H, m, Ph), 4.48 (2H, s, H- ), 3.63–3.58 (2H, m, H-
2387, 1611, 1460, 1354, 1147, 805 cmꢀ1; 1H NMR (D2O):
(8H, m, H- ,1,3,5), 3.06–3.02 (2H, m, H-8), 2.15–2.06 (2H, m, H-2),
1.83–1.71 (4H, m, H-6,7), 1.29 (3H, t, J¼7.3 Hz, H-
); 13C NMR (D2O):
46.0 (C- ), 13.4 (C- ), rest in Table 5; HRMS (ESI-MS): calcd for
d 3.18–3.08
a
a
b
8), 3.23–3.18 (2H, m, H-1), 3.16–3.11 (2H, m, H-5), 1.66–1.61 (2H, m,
d
a
b
H-2), 1.59–1.52 (2H, m, H-7), 1.44–1.36 (2H, m, H-6); 13C NMR
(MþH) C9H24N3 174.1970, found 174.1964.
(CDCl3):
133.5, 133.2, 133.2, 132.0 (2C), 131.9, 131.7, 131.0, 130.7, 128.8 (2C),
128.4 (2C),128.2,124.3,124.1,123.3 (2C) (totally 24C–Ph), 51.9 (C- ),
d 168.3 (2C, C]O), 148.0, 147.9, 135.6, 134.0 (2C), 133.7,
4.3.29. N1-(3-Ethylamino-1,1-2H2-propyl)-butane-1,4-diamine
trihydrochloride 2b
a
rest in Table 9; HRMS (ESI-MS): calcd for (MþH) C34H32D2N5O10S2
Prepared as 2a from 12b (800 mg, 1.18 mmol) to give 2b
(172 mg, 51%, 99% d) as a colourless solid, mp >308 ꢁC decomp. IR
(KBr): 3000–2264, 1612, 1460, 1346, 1144, 844, 803 cmꢀ1; 1H NMR
738.1873, found 738.1840.
4.3.26. 2-(4-{[3-(Isopropyl-(2-nitrobenzenesulfonyl)amino)-
propyl]-(2-nitrobenzenesulfonyl)amino}-butyl)isoindole-1,3-
dione 12f
(D2O):
2.07 (2H, m, H-2), 1.84–1.72 (4H, m, H-6,7), 1.30 (3H, t, J¼7.3 Hz, H-
); 13C NMR (D2O):
46.0 (C- ), 13.4 (C- ), rest in Table 5; HRMS
d 3.17–3.09 (6H, m, H-a,1,5), 3.08–3.02 (2H, m, H-8), 2.13–
b
d
a
b
Prepared as 12a from 11g (973 mg, 2.0 mmol) to give 12f
(1.28 g, 93%) as a colourless solid, mp 46–54 ꢁC (amorphous solid).
Rf 0.30 (EtOAc–hexane 2:1); IR (KBr): 3096, 2940 (br), 1710, 1543,
(ESI-MS): calcd for (MþH) C9H22N3D2 176.2096, found 176.2092.
Isotope distribution: 0% d0, 1% d1, 99% d2.
1373, 1159, 721 cmꢀ1
;
1H NMR (CDCl3):
d
8.04–7.99 (2H, m, Ph),
7.85–7.80 (2H, m, Ph), 7.74–7.62 (6H, m, Ph), 7.60–7.54 (2H, m, Ph),
4.12 (1H, sep., J¼6.7 Hz, H- ), 3.69–3.63 (2H, m, H-8), 3.39–3.32
(4H, m, H-3,5), 3.23–3.17 (2H, m, H-1), 1.96–1.88 (2H, m, H-2),
1.69–1.54 (4H, m, H-6,7), 1.12 (6H, d, J¼6.7 Hz, H-
); 13C NMR
(CDCl3): 168.3 (2C, C]O), 148.1, 148.0, 134.0 (2C), 133.6, 133.5,
133.4, 133.4, 132.1 (2C), 131.8, 131.7, 130.9, 130.8, 124.1, 124.0, 123.3
(2C) (totally 18C–Ph), 50.1 (C- ), 21.3 (2 C- ), rest in Table 9;
4.3.30. N1-(3-Ethylamino-1,1,2,2-2H4-propyl)-butane-1,4-diamine
trihydrochloride 2c
a
Prepared as 2a from 12c (927 mg, 1.37 mmol) to give 2c (315 mg,
80%, 98% d) as a colourless solid, mp >285 ꢁC decomp. IR (KBr):
b
3000–2312, 1611, 1458, 1343, 1123, 823, 806 cmꢀ1 1H NMR (D2O):
;
d
d
3.18–3.09 (6H, m, H-
m, H-6,7), 1.30 (3H, t, J¼7.3 Hz, H-
13.4 (C-
a
,1,5), 3.08–3.02 (2H, m, H-8), 1.85–1.73 (4H,
b
); 13C NMR (D2O):
d
45.9 (C- ),
a
a
b
b
), rest in Table 5; HRMS (ESI-MS): calcd for (MþH)
HRMS (ESI-MS): calcd for (MþH) C30H34N5O10S2 688.1747, found
C9H20N3D4 178.2221, found 178.2217. Isotope distribution: 0%
688.1751.
d0–d2, 8% d3, 92% d4.
4.3.27. 2-(4-{[3-(Cyclohexyl-(2-nitrobenzenesulfonyl)amino)-
propyl]-(2-nitrobenzenesulfonyl)amino}butyl)isoindole-1,3-
dione 12g
4.3.31. N1-(3-Benzylamino-propyl)-butane-1,4-diamine
trihydrochloride 2d41
Prepared as 2a from 12d (1.24 g, 1.69 mmol) to give 2d (393 mg,
Prepared as 12a from 11h (1.05 g, 2.0 mmol) to give 12g (1.35 g,
93%) as a colourless solid, mp 53–63 ꢁC (amorphous solid). Rf 0.56
(EtOAc–hexane 3:2); IR (KBr): 3095, 2935, 2858, 1711, 1544, 1157,
68%) as a colourless solid, mp >300 ꢁC decomp. (lit. mp >300 ꢁC).41
IR (KBr): 3050–2417, 1610, 1448, 744, 698 cmꢀ1 1H NMR (D2O):
;
d
7.54–7.47 (5H, m, Ph), 4.28 (2H, s, H-
3.07–3.02 (2H, m, H-8), 2.17–2.07 (2H, m, H-2), 1.82–1.71 (4H, m, H-
6,7); 13C NMR (D2O):
133.4, 132.8 (2C), 132.7, 132.3 (2C) (totally
6C–Ph), 54.2 (C-
a), 3.22–3.08 (6H, m, H-1,3,5),
721 cmꢀ1; 1H NMR (CDCl3):
d
8.04–7.98 (2H, m, Ph), 7.85–7.80 (2H,
m, Ph), 7.74–7.62 (6H, m, Ph), 7.59–7.54 (2H, m, Ph), 3.73–3.63 (3H,
m, H-7, ), 3.39–3.31 (4H, m, H-3,5), 3.25–3.18 (2H, m, H-1), 1.95–
1.86 (2H, m, H-2), 1.79–1.71 (2H, m, H-geq), 1.69–1.54 (7H, m, H-
6,7,beq deq), 1.39–1.25 (4H, m, H-bax gax), 1.10–0.99 (1H, m, H-dax);
13C NMR (CDCl3):
168.3 (2C, C]O), 148.0, 148.0, 134.0 (2C), 133.9,
133.5, 133.4, 133.4, 132.1 (2C), 131.8, 131.6, 130.8, 130.7, 124.1, 124.0,
123.3 (2C) (totally 18C–Ph), 58.3 (C- ), 31.9 (2C- ), 26.0 (2C- ), 25.2
(C-
), rest in Table 9; HRMS (ESI-MS): calcd for (MþNa)
C33H37N5O10S2Na 750.1880, found 750.1867.
d
a
a
), rest in Table 5; HRMS (ESI-MS): calcd for (MþH)
C14H26N3 236.2127, found 236.2121.
,
,
d
4.3.32. N1-(3-Benzylamino-1,1-2H2-propyl)-butane-1,4-diamine
trihydrochloride 2e
a
b
g
Prepared as 2a from 12e (1.81 g, 2.45 mmol) to give 2e (542 mg,
64%, 99% d) as a colourless solid, mp >300 ꢁC decomp. IR (KBr):
d
3050–2380, 1611, 1447, 843, 744, 698 cmꢀ1
;
1H NMR (D2O):

558
M.R. Ha¨kkinen et al. / Tetrahedron 65 (2009) 547–562
d
7.55–7.48 (5H, m, Ph), 4.29 (2H, s, H-
3.14–3.09 (2H, m, H-5), 3.08–3.03 (2H, m, H-8), 2.16–2.10 (2H, m, H-
2), 1.84–1.72 (4H, m, H-6,7); 13C NMR (D2O):
133.3, 132.7 (2C),
132.6, 132.2 (2C) (totally 6C–Ph), 54.1 (C- ), rest in Table 5; HRMS
a
), 3.22–3.17 (2H, m, H-1),
4.3.38. 3-[(4-Iodobutyl)-(2-nitrobenzenesulfonyl)amino]propyl}-
carbamic acid tert-butyl ester 10b
d
A mixture of 10a (8.6 g, 19.1 mmol) and NaI (28.7 g, 191 mmol)
in acetone (250 mL) was heated at reflux with stirring for 48 h. The
precipitate was filtered off and the filtrate was evaporated to dry-
ness in vacuo. The residue was dissolved in EtOAc, washed with
water (250 mL) and brine (250 mL), and dried over MgSO4. Solvent
was evaporated in vacuo and the residue was purified on silica gel
using EtOAc–hexane 1:1 as an eluent, to give 10b (7.3 g, 70%) as
a yellow oil. Rf 0.54 (EtOAc–hexane 1:1); IR (neat): 3422 (br), 2975,
a
(ESI-MS): calcd for (MþH) C14H24N3D2 238.2252, found 238.2241.
Isotope distribution: 0% d0, 3% d1, 97% d2.
4.3.33. N1-(3-Isopropylamino-propyl)-butane-1,4-diamine
trihydrochloride 2f
Prepared as 2a from 12f (1.26 g, 1.84 mmol), but the stirring was
continued at room temperature for 70 h to give 2f (438 mg, 80%) as
a colourless solid, mp 236–239 ꢁC. IR (KBr): 3000–2361, 1606, 1457,
2935, 1701, 1366, 1162, 748, 584 cmꢀ1; 1H NMR (CDCl3):
d 8.03–7.98
(1H, m, Ph), 7.74–7.67 (2H, m, Ph), 7.66–7.61 (1H, m, Ph), 4.78 (1H,
br, NH), 3.40–3.29 (4H, m, H-3,5), 3.19–3.11 (4H, m, H-1,8),1.81–1.72
(4H, m, H-2,7), 1.69–1.61 (2H, m, H-6), 1.44 (9H, s, Me3C); 13C NMR
(CDCl3): d 156.0 (C]O), 148.1, 133.6, 133.3, 131.7, 130.7, 124.3 (totally
6C–Ph), 79.3 (O–C), 28.4 (3C–Me3), rest in Table 9; HRMS (ESI-MS):
1387, 1099, 964 cmꢀ1; 1H NMR (D2O):
a
d
3.44 (1H, sep., J¼6.5 Hz, H-
), 3.22–3.02 (8H, m, H-1,3,5,8), 2.15–2.05 (2H, m, H-2), 1.84–1.71
(4H, m, H-6,7), 1.33 (6H, d, J¼6.5 Hz, H-
b
); 13C NMR (D2O):
d
54.0 (C-
a
), 21.1 (2C-
b), rest in Table 5; HRMS (ESI-MS): calcd for (MþH)
C10H26N3 188.2127, found 188.2123.
calcd for (MþNa) C18H28IN3O6SNa 564.0641, found 564.0633.
4.3.34. N1-(3-Cyclohexylamino-propyl)-butane-1,4-diamine
trihydrochloride 2g
Prepared as 2f from 12g (1.33 g, 1.83 mmol) to give 2g (478 mg,
78%) as a colourless solid, mp 277–278 ꢁC. IR (KBr): 3000–2368,
4.3.39. {3-[{4-[{3-[Ethyl-(2-nitrobenzenesulfonyl)-amino]-propyl}-
(2-nitrobenzenesulfonyl)-amino]-butyl}-(2-nitrobenzenesulfonyl)-
amino]-propyl}-carbamic acid tert-butyl ester 13a
To a stirred mixture of 11a (1.0 g, 2.12 mmol) and K2CO3 (0.88 g,
6.35 mmol) in dry DMF (21 mL) was added 10b (1.26 g, 2.33 mmol)
and the reaction mixture was stirred for 17 h at room temperature.
The solids were filtered off, the filtrate was evaporated to dryness
in vacuo and the residue was purified on silica gel using first
EtOAc–hexane 2:1 and then 5:2 as an eluent, to give 13a (1.3 g, 69%)
as a colourless solid, mp 45–54 ꢁC (amorphous solid). Rf 0.35
(EtOAc–hexane 2:1); IR (KBr): 3422 (br), 3096, 2940 (br), 1707,
1607, 1456, 1050, 775 cmꢀ1; 1H NMR (D2O):
a
d
3.22–3.00 (9H, m, H-
,1,3,5,8), 2.15–2.02 (4H, m, H-2,beq), 1.89–1.71 (6H, m, H-6,7,geq),
1.70–1.63 (1H, m, H-deq), 1.40–1.26 (4H, m, H-bax gax), 1.24–1.12 (1H,
m, H-dax); 13C NMR (D2O):
60.4 (C- ), 31.8 (2C- ), 27.4 (C- ), 26.8
(2C-
,
d
a
b
d
g
), rest in Table 5; HRMS (ESI-MS): calcd for (MþH) C13H30N3
228.2440, found 228.2434.
4.3.35. N1-(3-Amino-1,1-2H2-propyl)butane-1,4-diamine
trihydrochloride 2h
1544, 1374, 1160, 744 cmꢀ1
Ph), 7.73–7.68 (6H, m, Ph), 7.64–7.59 (3H, m, Ph), 4.81 (1H, br, NH),
3.38–3.23 (12H, m, H- ,1,3,5,8,10), 3.16–3.09 (2H, m, H-12), 1.89–
1.81 (2H, m, H-2), 1.76–1.69 (2H, m, H-11), 1.54–1.47 (4H, m, H-6,7),
1.43 (9H, s, Me3C-), 1.11 (3H, t, J¼7.1 Hz, H-
); 13C NMR (CDCl3):
156.1 (C]O), 148.0 (2C), 148.0, 133.7, 133.7, 133.6, 133.3, 133.2,
133.0, 131.9, 131.9, 131.8, 130.8, 130.7, 130.6, 124.3, 124.2, 124.2
(totally 18C–Ph), 79.3 (O–C), 42.4 (C- ), 28.4 (3C–Me3), 13.6
(C-
; d 8.02–7.97 (3H, m,
1H NMR (CDCl3):
Prepared as 1l from 2e (300 mg, 0.87 mmol) to give 2h (122 mg,
55%, 99% d) as a colourless solid, mp 251–252 ꢁC (lit. mp for unla-
belled SPD: 250–254 ꢁC).48 IR (KBr): 3000–2370, 1596, 1451, 1171,
a
b
845 cmꢀ1
;
1H NMR (D2O):
d
3.14–3.02 (6H, m, H-1,5,8), 2.11–2.05
d
(2H, m, H-2), 1.83–1.72 (4H, m, H-6,7); 13C NMR (D2O):
d
in Table 5;
HRMS (ESI-MS): calcd for (MþH) C7H18N3D2 148.1783, found
148.1787. Isotope distribution: 0% d0, 1% d1, 99% d2.
a
b
), rest in Table 10; HRMS (ESI-MS): calcd for (MþNa)
C35H47N7O14S3Na 908.2241, found 908.2234.
4.3.36. [3-(2-Nitro-benzenesulfonylamino)-propyl]-carbamic acid
tert-butyl ester 9
4.3.40. {3-[{4-[{3-[Ethyl-(2-nitrobenzenesulfonyl)-amino]-1,1-2H2-
propyl}-(2-nitrobenzenesulfonyl)-amino]-butyl}-(2-nitrobenzene-
sulfonyl)-amino]-propyl}-carbamic acid tert-butyl ester 13b
Prepared as 13a from 11b (1.0 g, 2.11 mmol) to give 13b (1.40 g,
75%) as a colourless solid, mp 40–48 ꢁC (amorphous solid). Rf 0.35
(EtOAc–hexane 2:1); IR (KBr): 3424 (br), 3096–2880, 1708, 1544,
Prepared from (3-amino-propyl)-carbamic acid tert-butyl ester
(3.5 mL, 20 mmol), TEA (3.7 mL, 26.5 mmol) and NsCl (4.7 g,
21 mmol) as described earlier for (R)-N1-(o-nitrophenylsulfonyl)-
N3-(tert-butyloxycarbonyl)-1,3-diaminobutane32,33 to give 9 (7.0 g)
as yellow solid with 97% yield (lit.49 yield 89%). 1H and 13C NMR
chemical shift are in accordance with published data.50
1372, 1160, 748 cmꢀ1
7.73–7.67 (6H, m, Ph), 7.64–7.59 (3H, m, Ph), 4.83 (1H, br, NH), 3.38–
3.23 (10H, m, H- ,1,5,8,10), 3.16–3.09 (2H, m, H-12), 1.86–1.81 (2H,
m, H-2), 1.75–1.68 (2H, m, H-11), 1.54–1.47 (4H, m, H-6,7), 1.43 (9H,
); 13C NMR (CDCl3):
156.1
; d 8.02–7.96 (3H, m, Ph),
1H NMR (CDCl3):
4.3.37. 3-[(4-Chlorobutyl)-(2-nitrobenzenesulfonyl)amino]propyl}-
carbamic acid tert-butyl ester 10a
a
To a stirred mixture of 9 (7.2 g, 20 mmol) and K2CO3 (8.9 g,
64 mmol) in dry DMF (170 mL), 1-bromo-4-chlorobutane (14.9 mL,
129 mmol) was added and stirring was continued for 65 h at room
temperature. The precipitates were filtered off, the filtrate was
evaporated to dryness in vacuo, and the residue was suspensed in
EtOAc, washed with water (350 mL), brine (350 mL) and dried
over MgSO4. The solvent was evaporated in vacuo to give 10a
(8.9 g, 99% yield based on 9) as a pale yellow oil. IR (neat): 3423
s, Me3C–), 1.11 (3H, t, J¼7.1 Hz, H-
b
d
(C]O), 148.0 (2C), 148.0, 133.7 (2C), 133.6, 133.3 133.2, 133.0, 131.9,
131.9, 131.8, 130.8, 130.7, 130.6, 124.3, 124.2, 124.2 (totally 18C–Ph),
79.2 (O–C), 42.4 (C-a), 28.4 (3C–Me3), 13.6 (C-b), rest in Table 10;
HRMS (ESI-MS): calcd for (MþNa) C35H45D2N7O14S3Na 910.2366,
found 910.2392.
4.3.41. {3-[{4-[{3-[Benzyl-(2-nitrobenzenesulfonyl)-amino]-propyl}-
(2-nitrobenzenesulfonyl)-amino]-butyl}-(2-nitrobenzenesulfonyl)-
amino]-propyl}-carbamic acid tert-butyl ester 13c
;
(br), 3095–2850, 1701, 1365, 1163, 746 cmꢀ1 1H NMR (CDCl3):
d
8.03–7.98 (1H, m, Ph), 7.74–7.66 (2H, m, Ph), 7.65–7.61 (1H, m,
Ph), 4.78 (1H, br, NH), 3.54–3.49 (2H, m, H-8), 3.38–3.31 (4H, m, H-
3,5), 3.18–3.12 (2H, m, H-1), 1.79–1.66 (6H, m, H-2,6,7), 1.44 (9H, s,
Prepared as 13a from 11d (992 mg, 1.86 mmol) to give 13c
(1.42 g, 80%) as a colourless solid, mp 52–58 ꢁC (amorphous solid).
Rf 0.58 (EtOAc–hexane 3:1); IR (KBr): 3422 (br), 3096–2880, 1707,
Me3C); 13C NMR (CDCl3):
d 156.0 (C]O), 148.1, 133.6, 133.3, 131.7,
130.7, 124.3 (totally 6C–Ph), 79.3 (O–C), 28.4 (3C–Me3), rest in
Table 9; HRMS (ESI-MS): calcd for (MþNa) C18H28ClN3O6SNa
472.1285, found 472.1283.
1543, 1161, 742 cmꢀ1
;
1H NMR (CDCl3):
d
8.00–7.90 (3H, m, Ph),
7.74–7.56 (9H, m, Ph), 7.33–7.22 (5H, m, Ph), 4.81 (1H, br, NH), 4.47
(2H, s, H- ), 3.32–3.03 (12H, m, H-1,3,5,8,10,12), 1.74–1.67 (2H, m,
a

M.R. Ha¨kkinen et al. / Tetrahedron 65 (2009) 547–562
559
H-11), 1.66–1.57 (2H, m, H-2), 1.48–1.30 (4H, m, H-6,7), 1.43 (9H, s,
Me3C–); 13C NMR (CDCl3):
156.0 (C]O), 148.0, 148.0, 147.9, 135.5,
133.7, 133.7, 133.6, 133.2, 133.1, 133.0, 131.9, 131.9 (2C), 130.9, 130.7,
130.6, 128.8 (2C), 128.4 (2C), 128.2, 124.3, 124.2, 124.2 (totally 24C–
at room temperature and then evaporated to dryness in vacuo. The
residue was dissolved in a mixture of DCM and 2 M KOH (4:3,
130 mL), the water layer was extracted with DCM (4ꢂ50 mL), and
the combined organic extracts were dried over MgSO4. The solvent
was evaporated in vacuo, the residue was dissolved in 1,4-dioxane
(12 mL) followed with concd HCl (3 mL) and the resulting mixture
was stirred for 2 h at room temperature. The reaction mixture was
evaporated to dryness in vacuo and co-evaporated once with di-
oxane (10 mL). DCM was added to the residue, the product was
filtered and washed with DCM and cold ethanol. Recrystallization
from ethanol–water–EtOAc yielded 3a (269 mg, 50%) as a colour-
less solid, mp >275 ꢁC decomp. IR (KBr): 3000–2388, 1612, 1461,
d
Ph), 79.3 (O–C), 52.0 (C-a), 28.4 (3C–Me3), rest in Table 10; HRMS
(ESI-MS): calcd for (MþNa) C40H49N7O14S3Na 970.2397, found
970.2354.
4.3.42. {3-[{4-[{3-[Benzyl-(2-nitrobenzenesulfonyl)-amino]-
1,1,2,2-2H4-propyl}-(2-nitrobenzenesulfonyl)-amino]-butyl}-(2-
nitrobenzenesulfonyl)-amino]-propyl}-carbamic acid tert-butyl
ester 13d
Prepared as 13a from 11f (1.83 g, 3.39 mmol) to give 13d
(2.75 g, 85%) as a colourless solid, mp 44–54 ꢁC (amorphous
solid). Rf 0.58 (EtOAc–hexane 3:1); IR (KBr): 3423 (br), 3095–
1355, 1145, 806 cmꢀ1
,1,3,5,8,10,12), 2.16–2.06 (4H, m, H-2,11), 1.84–1.75 (4H, m, H-6,7),
); 13C NMR (D2O):
45.9 (C- ), 13.3 (C- ),
; d 3.21–3.08 (14H, m, H-
1H NMR (D2O):
a
1.30 (3H, t, J¼7.3 Hz, H-
b
d
a
b
2880, 1707, 1544, 1162, 743 cmꢀ1
;
1H NMR (CDCl3):
d
8.00–7.90
(3H, m, Ph), 7.74–7.56 (9H, m, Ph), 7.33–7.23 (5H, m, Ph), 4.82 (1H,
br, NH), 4.47 (2H, s, H- ), 3.32–3.05 (10H, m, H-1,5,8,10,12), 1.74–
1.66 (2H, m, H-11), 1.48–1.30 (4H, m, H-6,7) 1.43 (9H, s, Me3C–);
13C NMR (CDCl3):
156.0 (C]O), 148.0, 148.0, 147.9, 135.5, 133.8,
rest in Table 6; HRMS (ESI-MS): calcd for (MþH) C12H31N4 231.2549,
found 231.2555.
a
4.3.46. N-(3-Aminopropyl)-N0-(3-ethylamino-1,1-2H2-propyl)-
butane-1,4-diamine tetrahydrochloride 3b
d
133.7, 133.6, 133.2, 133.1, 133.0, 132.0, 131.9 (2C), 130.9, 130.7,
130.6, 128.8 (2C), 128.4 (2C), 128.2, 124.3, 124.2, 124.2 (totally
Prepared as 3a from 13b (1.0 g, 1.13 mmol) to give 3b (199 mg,
47%, 98% d) as a colourless solid, mp >300 ꢁC decomp. IR (KBr):
24C–Ph), 79.2 (O–C), 52.0 (C-
a
), 28.4 (3C–Me3), rest in Table 10;
3000–2382, 1611, 1460, 1354, 1145, 844 cmꢀ1 1H NMR (D2O):
;
HRMS (ESI-MS): calcd for (MþNa) C40H45D4N7O14S3Na 974.2648,
d
3.20–3.08 (12H, m, H-
1.84–1.76 (4H, m, H-6,7), 1.30 (3H, t, J¼7.3 Hz, H-
46.0 (C- ), 13.4 (C- ), rest in Table 6; HRMS (ESI-MS): calcd for
a
,1,5,8,10,12), 2.15–2.07 (4H, m, H-2,11),
found 974.2607.
b
); 13C NMR (D2O):
d
a
b
4.3.43. {3-[{4-[{3-[Isopropyl-(2-nitrobenzenesulfonyl)-amino]-
propyl}-(2-nitrobenzenesulfonyl)-amino]-butyl}-(2-nitrobenzene-
sulfonyl)-amino]-propyl}-carbamic acid tert-butyl ester 13e
Prepared as 13a from 11g (973 mg, 2.0 mmol) to give 13e (1.53 g,
85%) as a colourless solid, mp 50–55 ꢁC (amorphous solid). Rf 0.44
(EtOAc–hexane 2:1); IR (KBr): 3421 (br), 3097–2880, 1707, 1544,
(MþH) C12H29N4D2 233.2674, found 233.2666. Isotope distribution:
0% d0, 3% d1, 97% d2.
4.3.47. N-(3-Aminopropyl)-N0-(3-Benzylamino-propyl)-butane-
1,4-diamine tetrahydrochloride 3c52
Prepared as 3a from 13c (1.40 g, 1.48 mmol) to give 3c (405 mg,
62%) as a colourless solid, mp >300 ꢁC decomp. IR (KBr): 3000–
1161, 744 cmꢀ1; 1H NMR (CDCl3):
d
8.04–7.96 (3H, m, Ph), 7.74–7.66
(6H, m, Ph), 7.64–7.57 (3H, m, Ph), 4.84 (1H, br, NH), 4.10 (1H, sep.,
J¼6.7 Hz, H- ), 3.35–3.26 (8H, m, H-3,5,8,10), 3.22–3.09 (4H, m, H-
1,12), 1.93–1.85 (2H, m, H-2), 1.76–1.68 (2H, m, H-11), 1.55–1.46 (4H,
m, H-6,7), 1.43 (9H, s, Me3C-), 1.11 (6H, d, J¼6.7 Hz, H-
); 13C NMR
(CDCl3): 156.1 (C]O), 148.1, 148.0, 148.0, 133.7 (2C), 133.5, 133.5,
133.2, 133.2, 131.9, 131.9, 131.7, 130.8 (2C), 130.6, 124.3, 124.2, 124.1
(totally 18C–Ph), 79.2 (O–C), 50.1 (C- ), 28.4 (3C–Me3), 21.3 (2C- ),
2420, 1609, 1460, 874, 744, 698 cmꢀ1
(5H, m, Ph), 4.28 (2H, s, H-a
;
1H NMR (D2O):
d
7.55–7.47
), 3.25–3.05 (12H, m, H-1,3,5,8,10,12),
2.20–2.05 (4H, m, H-2,11), 1.85–1.73 (4H, m, H-6,7); 13C NMR (D2O):
133.3, 132.7 (2C), 132.7, 132.2 (2C) (totally 6C–Ph), 54.1 (C- ), rest
a
b
d
a
d
in Table 6; HRMS (ESI-MS): calcd for (MþH) C17H33N4 293.2705,
found 293.2698.
a
b
rest in Table 10; HRMS (ESI-MS): calcd for (MþNa)
4.3.48. N-(3-Aminopropyl)-N0-(3-Benzylamino-1,1,2,2-2H4-propyl)-
butane-1,4-diamine tetrahydrochloride 3d
C36H49N7O14S3Na 922.2397, found 922.2361.
Prepared as 3a from 13d (2.73 g, 2.86 mmol) to give 3d
(904 mg, 71%, 98% d) as a colourless solid, mp >300 ꢁC decomp. IR
(KBr): 3050–2309, 1610, 1450, 825, 744, 698 cmꢀ1; 1H NMR (D2O):
4.3.44. {3-[{4-[{3-[Cyclohexyl-(2-nitrobenzenesulfonyl)-amino]-
propyl}-(2-nitrobenzenesulfonyl)-amino]-butyl}-(2-nitrobenzene-
sulfonyl)-amino]-propyl}-carbamic acid tert-butyl ester 13f
Prepared as 13a from 11h (1.05 g, 2.0 mmol) to give 13f (1.19 g,
63%) as a colourless solid, mp 54–63 ꢁC (amorphous solid). Rf 0.44
(EtOAc–hexane 2:1); IR (KBr): 3442 (br), 3096, 2936, 2861, 1707,
d
7.55–7.47 (5H, m, Ph), 4.28 (2H, s, H-
1,5,8,10,12), 2.17–2.05 (2H, m, H-11), 1.85–1.73 (4H, m, H-6,7);13C
NMR (D2O): 133.3, 132.7 (2C), 132.7, 132.2 (2C) (totally 6C–Ph),
54.1 (C-
a), 3.25–3.05 (10H, m, H-
d
a
), rest in Table 6; HRMS (ESI-MS): calcd for (MþH)
1544, 1159, 742 cmꢀ1
;
1H NMR (CDCl3):
d
8.04–7.95 (3H, m, Ph),
7.74–7.66 (6H, m, Ph), 7.64–7.56 (3H, m, Ph), 4.84 (1H, br, NH), 3.70–
3.62 (1H, m, H- ), 3.37–3.25 (8H, m, H-3,5,8,10), 3.23–3.17 (2H, m,
H-1), 3.16–3.09 (2H, m, H-12),1.91–1.84 (2H, m, H-2),1.78–1.68 (4H,
m, H-11, geq), 1.66–1.57 (3H, m, H-beq deq), 1.54–1.44 (4H, m, H-6,7),
1.43 (9H, s, Me3C–), 1.38–1.24 (4H, m, H-bax gax), 1.10–0.99 (1H, m,
H-dax); 13C NMR (CDCl3):
156.0 (C]O), 148.1, 148.0, 148.0, 133.8,
133.7 (2C), 133.5, 133.2, 133.2, 131.9, 131.9, 131.7, 130.9, 130.6, 130.6,
124.3, 124.2, 124.0 (totally 18C–Ph), 79.2 (O–C), 58.4 (C- ), 31.9 (2C-
), 28.4 (3C–Me3), 26.0 (2C- ), 25.2 (C- ), rest in Table 10; HRMS
C17H29N4D4 297.2956, found 297.2947. Isotope distribution: 0% d0–
d2, 7% d3, 92% d4.
a
4.3.49. N-(3-Aminopropyl)-N0-(3-isopropylaminopropyl)butane-
1,4-diamine tetrahydrochloride 3e
Prepared as 3a from 13e (1.51 g, 1.68 mmol) to give 3e (452 mg,
69%) as a colourless solid, mp >269 ꢁC decomp. IR (KBr): 3000–
,
,
d
2364, 1625, 1460, 1376, 877 cmꢀ1
;
1H NMR (D2O):
d
3.44 (1H, sep.,
), 3.22–3.00 (12H, m, H-1,3,5,8,10,12), 2.18–2.00 (4H,
m, H-2,11), 1.85–1.70 (4H, m, H-6,7), 1.33 (6H, d, J¼6.5 Hz, H-
); 13
NMR (D2O): 53.9 (C- ), 21.0 (2C- ), rest in Table 6; HRMS (ESI-
MS): calcd for (MþH) C13H33N4 245.2705, found 245.2695.
a
J¼6.5 Hz, H-
a
b
g
d
b
C
(ESI-MS): calcd for (MþNa) C39H53N7O14S3Na 962.2710, found
d
a
b
962.2683.
4.3.45. N-(3-Aminopropyl)-N0-(3-ethylamino-propyl)butane-1,4-
diamine tetrahydrochloride 3a51
4.3.50. N-(3-Aminopropyl)-N0-(3-cyclohexylaminopropyl)butane-
1,4-diamine tetrahydrochloride 3f
A mixture of 13a (1.27 g, 1.43 mmol), PhSH (0.67 mL, 6.48 mmol)
and K2CO3 (1.78 g, 12.9 mmol) in DMF (13 mL) was stirred for 20 h
Prepared as 3a from 13f (1.15 g, 1.22 mmol) to give 3f (380 mg,
72%) as
colourless solid, mp >285 ꢁC decomp. IR (KBr):
a

560
M.R. Ha¨kkinen et al. / Tetrahedron 65 (2009) 547–562
3000–2419, 1610, 1460, 1051, 775 cmꢀ1
;
1H NMR (D2O):
d
3.21–
4.3.55. N,N0-Bis-[3-(cyclohexyl-{2-nitrobenzenesulfonyl}-
3.07 (13H, m, H-
1.89–1.74 (6H, m, H-6,7,geq), 1.71–1.64 (1H, m, H-deq), 1.40–1.26
(4H, m, H-bax 60.4
gax), 1.23–1.13 (1H, m, H-dax); 13C NMR (D2O):
(C- ), 31.7 (2C- ), 27.3 (C- ), 26.7 (2C- ), rest in Table 6; HRMS
a
,1,3,5,8,10,12), 2.15–2.02 (6H, m, H-2,11,beq),
amino)propyl]-N,N0-bis-(2-nitrobenzenesulfonyl)butane-1,4-
diamine 14d
,
d
To a stirred mixture of 11h (1.18 g, 2.24 mmol) and K2CO3
(0.88 g, 6.39 mmol) in dry DMF (12 mL), 1,4-diiodobutane (330 mg,
1.06 mmol) was added and stirring was continued for 24 h at room
temperature. Solids were filtered off and the filtrate was evaporated
to dryness in vacuo. The residue was treated with EtOAc (25 mL)
and water (25 mL), the water layer was extracted with EtOAc
(2ꢂ25 mL), and the combined organic phases were washed with
brine (25 mL) and dried over MgSO4. Solvent was evaporated in
vacuo and the residue was purified on silica gel using first EtOAc–
hexane 3:2 and then 3:1 as an eluent, affording 14d (993 mg, 84%
yield based on 1,4-diiodobutane) as a colourless solid, mp 62–72 ꢁC
(amorphous solid). Rf 0.31 (EtOAc–hexane 3:2); IR (KBr): 3096,
a
b
d
g
(ESI-MS): calcd for (MþH) C16H37N4 285.3018, found 285.3009.
4.3.51. N-(3-Amino-1,1,2,2-2H4-propyl)-N0-(3-aminopropyl)-
butane-1,4-diamine tetrahydrochloride 4h
Prepared as 1l from 3d (400 mg, 0.90 mmol) to give 4h (262 mg,
82%, 98% d) as a colourless solid, mp >300 ꢁC decomp. IR (KBr):
3124–2010, 1594, 1479, 1158, 812 cmꢀ1; 1H NMR (D2O):
d
3.22–3.08
(10H, m, H-1,5,8,10,12), 2.16–2.07 (2H, m, H-11), 1.86–1.75 (4H, m,
H-6,7); 13C NMR (D2O):
in Table 6; HRMS (ESI-MS): calcd for
d
(MþH) C10H23N4D4 207.2487, found 207.2486. Isotope distribution:
0% d0–d2, 8% d3, 92% d4.
2935, 2858, 1543, 1158, 741 cmꢀ1; 1H NMR (CDCl3):
m, Ph), 7.74–7.66 (8H, m, Ph), 7.63–7.55 (4H, m, Ph), 3.72–3.63 (2H,
m, H- ), 3.35–3.26 (8H, m, H-3,5,8,10), 3.24–3.18 (4H, m, H-1,12),
1.92–1.83 (4H, m, H-2,11), 1.78–1.71 (4H, m, H-geq), 1.67–1.57 (6H,
m, H-beq deq), 1.52–1.45 (4H, m, H-6,7), 1.39–1.25 (8H, m, H-bax gax),
1.09–0.98 (2H, m, H-dax); 13C NMR (CDCl3):
148.1 (2C), 147.9 (2C),
d 8.04–7.97 (4H,
4.3.52. N,N0-Bis-[3-(ethyl{2-nitrobenzenesulfonyl}amino)propyl]-
N,N0-bis-(2-nitrobenzenesulfonyl)butane-1,4-diamine 14a
a
To a stirred mixture of 11a (894 mg, 1.89 mmol) and K2CO3
(0.75 g, 5.4 mmol) in dry DMF (10 mL), 1,4-diiodobutane (279 g,
0.90 mmol) was added and stirring was continued for 24 h at room
temperature. The solids were filtered off and the filtrate was
evaporated to dryness in vacuo. The residue was treated with EtOAc
(20 mL) and water (20 mL) and this resulted in precipitation of the
product, which was filtered off and dried in vacuo yielding 14a
(718 mg, 80%) as a sparingly soluble colourless solid, mp 151–
,
,
d
133.8 (2C), 133.7 (2C), 133.4 (2C), 133.2 (2C), 132.0 (2C), 131.7 (2C),
130.8 (2C), 130.6 (2C), 124.2 (2C), 124.0 (2C) (totally 24C–Ph), 58.4
(2C-a), 31.9 (4C-b), 26.0 (4C-g), 25.2 (2C-d), rest in Table 10; HRMS
(ESI-MS): calcd for (MþNa) C46H58N8O16S4Na 1129.2751, found
1129.2792.
156 ꢁC. IR (KBr): 3102–2850, 1542, 1376, 1161, 744 cmꢀ1
(CDCl3þDMSO-d6): 8.01–7.94 (4H, m, Ph), 7.79–7.71 (8H, m, Ph),
7.70–7.64 (4H, m, Ph), 3.39–3.23 (16H, m, H- ,1,3,5,8,10,12), 1.88–
;
1H NMR
4.3.56. N,N0-Bis-(3-ethylamino-propyl)butane-1,4-diamine
d
tetrahydrochloride 4a53
a
A mixture of 14a (700 mg, 0.70 mmol), PhSH (0.43 mL,
1.79 (4H, m, H-2,11), 1.55–1.48 (4H, m, H-6,7), 1.11 (6H, t, J¼7.1 Hz,
4.20 mmol) and K2CO3 (1.16 g, 8.41 mmol) in DMF (7 mL) was
stirred for 20 h at room temperature and then the reaction
mixture was evaporated to dryness in vacuo. The residue was
dissolved in a mixture of DCM and 2 M KOH (4:3, 60 mL), the
water layer was extracted with a mixture of ethanol and DCM
(1:2, 4ꢂ20 mL), and the combined organic extracts were dried
over MgSO4. Solvents were evaporated in vacuo, the residue was
dissolved in 1,4-dioxane (6 mL) followed by the addition of concd
HCl (1.5 mL) and the resulting mixture was stirred for 5 min at
room temperature. The reaction mixture was evaporated to
dryness in vacuo and co-evaporated once with dioxane (10 mL).
DCM was added to the residue and the product was filtered,
washed with DCM and cold ethanol. Recrystallization from eth-
anol–water–EtOAc yielded 4a (144 mg, 51%) as a colourless solid,
mp >300 ꢁC decomp. IR (KBr): 3000–2389, 1596, 1460, 1354,
H-b
); 13C NMR (CDCl3þDMSO-d6):
d 148.0 (2C), 148.0 (2C), 133.7
(2C), 133.6 (2C), 133.3 (2C), 132.9 (2C), 132.0 (2C), 131.9 (2C), 130.8
(2C), 130.6 (2C), 124.2 (2C), 124.2 (2C) (totally 24C–Ph), 42.4 (2C-
13.6 (2C-
C38H46N8O16S4K 1037.1552, found 1037.1588.
a),
b
), rest in Table 10; HRMS (ESI-MS): calcd for (MþK)
4.3.53. N,N0-Bis-[3-(ethyl-{2-nitro-benzenesulfonyl}-amino)-
1,1-2H2-propyl]-N,N0-bis-(2-nitro-benzenesulfonyl)butane-1,4-
diamine 14b
Prepared as 14a from 11b (1.50 g, 3.16 mmol) and 1,4-diiodo-
butane (467 mg, 1.51 mmol) to give 14b (1.07 g, 71%) as a colourless
solid, mp 155–158 ꢁC. IR (KBr): 3102, 2941, 1541, 1374, 1160 cmꢀ1
;
1H NMR (CDCl3þDMSO-d6):
d
8.01–7.94 (4H, m, Ph), 7.78–7.70 (8H,
,1,5,8,12),1.87–
m, Ph), 7.69–7.62 (4H, m, Ph), 3.39–3.23 (12H, m, H-
a
1.78 (4H, m, H-2,11), 1.56–1.46 (4H, m, H-6,7), 1.11 (6H, t, J¼7.1 Hz,
1142, 806 cmꢀ1
;
1H NMR (D2O) chemical shifts as reported ear-
45.9 (2C- ), 13.3 (2C- ), rest in Table 6;
H-
b
); 13C NMR (CDCl3þDMSO-d6):
d
147.9 (2C), 147.9 (2C), 133.8
lier;53 13C NMR (D2O):
d
a
b
(2C), 133.7 (2C), 133.0 (2C), 132.7 (2C), 132.0 (2C), 131.9 (2C), 130.5
HRMS (ESI-MS): calcd for (MþH) C14H35N4 259.2862, found
(2C), 130.4 (2C), 124.2 (2C), 124.1 (2C) (totally 24C–Ph), 42.4 (2C-
13.6 (2C-
C38H42D4N8O16S4K 1041.1803, found 1041.1773.
a
),
259.2854.
b
), rest in Table 10; HRMS (ESI-MS): calcd for (MþK)
4.3.57. N,N0-Bis-(3-ethylamino-1,1-2H2-propyl)butane-1,4-diamine
tetrahydrochloride 4b
Prepared as 4a from 14b (1.04 g, 1.04 mmol) to give 4b (190 mg,
45%, 99% d) as a colourless solid, mp >300 ꢁC decomp. IR (KBr):
3000–2265, 1595, 1461, 1348, 1145, 843, 803 cmꢀ1; 1H NMR (D2O):
4.3.54. N,N0-Bis-[3-(isopropyl-{2-nitrobenzenesulfonyl}amino)-
propyl]-N,N0-bis-(2-nitrobenzenesulfonyl)butane-1,4-diamine 14c
Prepared as 14a from 11g (3.08 g, 6.33 mmol) and 1,4-diiodo-
butane (934 mg, 3.02 mmol) to give 14c (2.68 g, 87%) as a col-
ourless solid, mp 202–204 ꢁC. IR (KBr): 3090–2850, 1544, 1362,
d
3.19–3.07 (12H, m, H-
1.74 (4H, m, H-6,7),1.30 (6H, t, J¼7.3 Hz, H-
(2C- ), 13.3 (2C-
a
,1,5,8,12), 2.15–2.06 (4H, m, H-2,11), 1.85–
b
); 13C NMR (D2O):
d
45.9
1157 cmꢀ1
;
1H NMR (CDCl3þDMSO-d6):
d
8.03–7.91 (8H, m, Ph),
), 3.31–3.22
a
b
), rest in Table 6; HRMS (ESI-MS): calcd for (MþH)
7.90–7.79 (8H, m, Ph), 3.97 (2H, sep., J¼6.7 Hz, H-
a
C14H31N4D4 263.3113, found 263.3103. Isotope distribution: 0% d0–
d2, 3% d3, 97% d4.
(8H, m, H-3,5,8,10), 3.15–3.09 (4H, m, H-1–12), 1.79–1.71 (4H, m,
H-2,11), 1.47–1.40 (4H, m, H-6,7), 1.02 (12H, d, J¼6.7 Hz, H-
); 13C
NMR (CDCl3þDMSO-d6): 147.4 (2C), 147.4 (2C), 134.4 (4C), 132.3
(2C), 132.2 (2C), 132.0 (2C), 131.7 (2C), 129.8 (4C), 124.2 (2C), 124.1
(2C) (totally 24C–Ph), 49.6 (2C- ), 20.6 (4C- ), rest in Table 10;
b
d
4.3.58. N,N0-Bis-(3-isopropylaminopropyl)butane-1,4-diamine
tetrahydrochloride 4c53
Prepared as 4a from 14c (1.03 g, 1.0 mmol), PhSH (0.64 mL,
6.2 mmol) and K2CO3 (1.66 g, 12 mmol) in DMF (10 mL), but stirring
was continued for 70 h to give 4c (282 mg, 65%) as a colourless
a
b
HRMS (ESI-MS): calcd for (MþK) C40H50N8O16S4K 1065.1865,
found 1065.1844.

M.R. Ha¨kkinen et al. / Tetrahedron 65 (2009) 547–562
561
solid, mp >281 ꢁC decomp. IR (KBr): 3000–2360, 1636, 1478, 1386,
1150, 762 cmꢀ1 1H NMR (D2O) chemical shifts as reported ear-
lier;53 13C NMR (D2O):
53.9 (2C- ), 21.0 (4C- ), rest in Table 6;
(neat): 3096, 2937 (br), 2224, 1541, 1772, 1160, 744 cmꢀ1
NMR (CDCl3):
7.65–7.58 (2H, m, Ph), 3.40–3.26 (6H, m, H-
J¼6.6 Hz, H-8), 1.83–1.74 (2H, m, H-7), 1.70–1.61 (2H, m, H-6),
1.12 (3H, t, J¼7.1 Hz, H- 148.0 (2C),
); 13C NMR (CDCl3):
133.7, 133.6, 133.3, 133.0, 131.9, 131.8, 130.8, 130.7, 124.2, 124.2
(totally 12C–Ph), 42.4 (C- ), 13.6 (C- ), rest in Table 9; HRMS
;
1H
;
d
8.04–7.97 (2H, m, Ph), 7.74–7.66 (4H, m, Ph),
,1,5), 3.17 (2H, t,
d
a
b
a
HRMS (ESI-MS): calcd for (MþH) C16H39N4 287.3175, found
287.3165.
b
d
4.3.59. N,N0-Bis-(3-cyclohexylaminopropyl)butane-1,4-diamine
tetrahydrochloride 4d54
Prepared as 4c from 14d (968 mg, 0.87 mmol) to give 4d
(364 mg, 82%) as colourless solid, mp >300 ꢁC decomp. IR (KBr):
a
b
(ESI-MS): calcd for (MþH) C21H24D4IN4O8S2 659.0644, found
659.0656.
3000–2420,1596,1456,1052, 775 cmꢀ1; 1H NMR (D2O):
d
3.21–3.07
,1,3,5,8,10,12), 2.15–2.02 (8H, m, H-2,11,beq), 1.89–1.74
(8H, m, H-6,7,geq), 1.71–1.63 (2H, m, H-deq), 1.40–1.26 (8H, m, H-
bax 60.4 (2C, C- ),
gax), 1.24–1.12 (2H, m, H-dax); 13C NMR (D2O):
31.7 (4C, C- ), 27.3 (2C, C- ), 26.7 (4C, C- ), rest in Table 6; HRMS
4.3.64. N-[3-(Benzyl-{2-nitrobenzenesulfonyl}amino)-propyl]-N0-
[3-(ethyl-{2-nitro-benzenesulfonyl}amino)-propyl]-N,N0-bis-{2-
nitrobenzenesulfonyl}butane-1,4-diamine 14e
(14H, m, H-
a
,
d
a
To a stirred mixture of 11d (535 mg, 1.0 mmol) and K2CO3
(414 mg, 3.0 mmol) in dry DMF (10 mL), 15c (720 mg, 1.1 mmol)
was added and stirring was continued for 24 h at room temper-
ature. Solids were filtered off, the filtrate was evaporated to
dryness in vacuo, and the precipitate was dissolved in EtOAc
(20 mL) and water (20 mL). The water layer was extracted with
EtOAc (20 mL) and the combined organic extracts were washed
with brine (20 mL) and dried over MgSO4. After evaporation of
the solvent in vacuo, the residue was purified on silica gel using
first EtOAc–hexane 3:2 and then 3:1 as the eluent, to give 14e
(939 mg, 88%) as a colourless solid, mp 45–56 ꢁC (amorphous
solid). Rf 0.44 (EtOAc–hexane 3:1); IR (KBr): 3096, 2940 (br),
b
d
g
(ESI-MS): calcd for (MþH) C22H47N4 367.3801, found 367.3785.
4.3.60. N1-(4-Chlorobutyl)-N3-ethyl-N,N3-bis-(2-
nitrobenzenesulfonyl)propane-1,3-diamine 15a
To a stirred mixture of 11a (2.02 g, 4.26 mmol) and K2CO3
(1.76 g, 12.77 mmol) in dry DMF (34 mL) was added 1-bromo-4-
chlorobutane (2.94 mL, 25.55 mmol) and stirring was continued
for 65 h at room temperature. Solids were filtered off and the
filtrate was evaporated to dryness in vacuo. The residue was
treated with EtOAc, washed with water (35 mL), brine (35 mL)
and dried over MgSO4. Solvent was evaporated in vacuo affording
15a (2.45 g, w100%) as a yellow oil. IR (neat): 3096, 2940 (br),
1543, 1374, 1160, 744 cmꢀ1
Ph), 7.74–7.57 (12H, m, Ph), 7.33–7.23 (5H, m, Ph), 4.47 (2H, s,
Ph–CH2–), 3.35 (2H, q, J¼7.1 Hz, H- ), 3.30–3.04 (12H, m, H-
1,3,5,8,10,12), 1.88–1.80 (2H, m, H-11), 1.67–1.59 (2H, m, H-2),
1.47–1.31 (4H, m, H-6,7), 1.10 (3H, t, J¼7.1 Hz, H-
); 13C NMR
(CDCl3): 148.0, 148.0, 148.0, 147.9, 135.5, 133.7, 133.7, 133.7,
; d 8.02–7.90 (4H, m,
1H NMR (CDCl3):
1544, 1159, 745 cmꢀ1
7.73–7.66 (4H, m, Ph), 7.64–7.59 (2H, m, Ph), 3.55–3.51 (2H, m, H-
8), 3.41–3.27 (8H, m, H- ,1,3,5), 1.94–1.86 (2H, m, H-2), 1.80–1.66
); 13C NMR (CDCl3):
148.0 (2C), 133.7, 133.5, 133.3, 133.0, 131.9, 131.8, 130.9, 130.8,
;
1H NMR (CDCl3):
d
8.04–7.98 (2H, m, Ph),
a
a
b
(4H, m, H-6,7), 1.12 (3H, t, J¼7.1 Hz, H-
b
d
d
133.6, 133.3, 133.1, 133.0 (2C), 132.0, 132.0, 131.9, 131.8, 130.8,
130.8, 130.7, 130.6, 128.8 (2C), 128.4 (2C), 128.2, 124.3, 124.2 (2C),
124.2, 124.2 (totally 12C–Ph), 42.3 (C-a), 13.6 (C-b), rest in Table 9;
HRMS (ESI-MS): calcd for (MþH) C21H28ClN4O8S2 563.1037, found
124.2 (totally 30C–Ph), 52.0 (Ph–CH2–), 42.4 (C-a), 13.6 (C-b), rest
563.1038.
in Table 10; HRMS (ESI-MS): calcd for (MþNa) C43H48N8O16S4Na
1083.1969, found 1083.2011.
4.3.61. N1-(4-Chlorobutyl)-N3-ethyl-1,1,2,2-2H4-N1,N3-bis-(2-
nitrobenzenesulfonyl)propane-1,3-diamine 15b
Prepared as 15a from 11c (953 mg, 2.0 mmol) with subsequent
purification on silica gel using EtOAc–hexane 3:2 as an eluent to
give 15b (1.07 g, 95%) as a pale yellow oil. IR (neat): 3096, 2939 (br),
4.3.65. N-[3-(Benzyl-{2-nitrobenzenesulfonyl}amino)-1,1,2,2-2H4-
propyl]-N0-[3-(ethyl-{2-nitro-benzenesulfonyl}amino)propyl]-N,N0-
bis-{2-nitrobenzenesulfonyl}butane-1,4-diamine 14f
Prepared as 14e from 11f (1.62 g, 3.0 mmol) and 15c (2.16 g,
3.3 mmol) to give 14f (2.67 g, 83%) as a colourless solid, mp
44–56 ꢁC (amorphous solid). Rf 0.39 (EtOAc–hexane 2:1); IR
2226, 1544, 1161, 745 cmꢀ1
;
1H NMR (CDCl3):
d
8.05–7.96 (2H, m,
Ph), 7.74–7.65 (4H, m, Ph), 7.65–7.57 (2H, m, Ph), 3.55–3.51 (2H, m,
H-8), 3.42–3.25 (6H, m, H- ,1,5), 1.80–1.65 (4H, m, H-6,7), 1.12 (3H,
t, J¼7.1 Hz, H- 148.0 (2C), 133.7, 133.6, 133.3,
); 13C NMR (CDCl3):
133.1,131.9,131.8,130.8,130.7,124.2,124.2 (totally 12C–Ph), 42.4 (C-
), 13.6 (C-
a
(KBr): 3096, 2938 (br), 1544, 1373, 1160, 743 cmꢀ1 1H NMR
;
b
d
(CDCl3):
7.23 (5H, m, Ph), 4.47 (2H, s, Ph–CH2–), 3.35 (2H, q, J¼7.1 Hz,
H- ), 3.31–3.21 (6H, m, H-8,10,12), 3.19 (2H, s, H-1), 3.14–3.07
(2H, m, H-5), 1.88–1.79 (2H, m, H-11), 1.46–1.30 (4H, m, H-6,7),
); 13C NMR (CDCl3):
148.0, 148.0,
d 8.02–7.89 (4H, m, Ph), 7.75–7.57 (12H, m, Ph), 7.33–
a
b
), rest in Table 9; HRMS (ESI-MS): calcd for (MþH)
a
C21H24D4ClN4O8S2 567.1288, found 567.1302.
1.10 (3H, t, J¼7.1 Hz, H-
b
d
4.3.62. N1-(4-Iodobutyl)-N3-ethyl-N1,N3-bis-(2-
nitrobenzenesulfonyl)-propane-1,3-diamine 15c
148.0, 147.9, 135.5, 133.8, 133.7, 133.7, 133.6, 133.3, 133.1, 133.0,
133.0, 132.0, 132.0, 132.0, 131.9, 130.8, 130.8, 130.6, 130.6, 128.8
(2C), 128.4 (2C), 128.2, 124.3, 124.2 (2C), 124.2 (totally 30C–Ph),
Prepared as 10b from 15a (2.4 g, 4.26 mmol) to give 15c (2.49 g,
89%) as a slightly yellowish oil. Rf 0.44 (EtOAc–hexane 3:2); IR
52.0 (Ph–CH2–), 42.4 (C-a), 13.6 (C-b), rest in Table 10; HRMS
(neat): 3096, 2939 (br), 1542, 1159, 744 cmꢀ1
;
1H NMR (CDCl3):
(ESI-MS): calcd for (MþK) C43H44D4N8O16S4K 1103.1959, found
d
8.04–7.98 (2H, m, Ph), 7.74–7.66 (4H, m, Ph), 7.65–7.58 (2H, m, Ph),
1103.1908.
3.40–3.26 (8H, m, H-
a
,1,3,5), 3.17 (2H, t, J¼6.6 Hz, H-8), 1.95–1.86
(2H, m, H-2), 1.83–1.75 (2H, m, H-7), 1.70–1.62 (2H, m, H-6), 1.12
(3H, t, J¼7.1 Hz, H- 148.0 (2C), 133.7, 133.6,
); 13C NMR (CDCl3):
133.4, 133.0, 131.9, 131.8, 130.9, 130.8, 124.2, 124.2 (totally 12C–Ph),
42.4 (C- ), 13.6 (C- ), rest in Table 9; HRMS (ESI-MS): calcd for
4.3.66. N-[3-(Benzyl-{2-nitro-benzenesulfonyl}amino)-1,1,2,2-2H4-
propyl]-N0-[3-(ethyl-{2-nitro-benzenesulfonyl}amino)-1,1,2,2-2H4-
propyl]-N,N0-bis-{2-nitrobenzenesulfonyl}butane-1,4-diamine 14g
Prepared as 14e from 11f (792 mg, 1.47 mmol) and 15d (1.07 g,
1.62 mmol) to give 14g (1.32 g, 84%) as a colourless solid, mp 43–
54 ꢁC (amorphous solid). Rf 0.39 (EtOAc–hexane 2:1); IR (KBr):
b
d
a
b
(MþH) C21H28IN4O8S2 655.0393, found 655.0393.
4.3.63. N1-(4-Iodobutyl)-N3-ethyl-1,1,2,2-2H4-N1,N3-bis-(2-
nitrobenzenesulfonyl)propane-1,3-diamine 15d
Prepared as 10b from 15b (1.04 g, 1.84 mmol) to give 15d
(1.15 g, 95%) as a yellowish oil. Rf 0.44 (EtOAc–hexane 3:2); IR
3096, 2937 (br), 1541, 1373, 1161, 743 cmꢀ1
8.01–7.89 (4H, m, Ph), 7.74–7.56 (12H, m, Ph), 7.32–7.23 (5H,
m, Ph), 4.47 (2H, s, Ph–CH2–), 3.34 (2H, q, J¼7.1 Hz, H- ), 3.27
(2H, s, H-12), 3.25–3.21 (2H, m, H-8), 3.19 (2H, s, H-1), 3.13–3.07
;
1H NMR (CDCl3):
d
a

562
M.R. Ha¨kkinen et al. / Tetrahedron 65 (2009) 547–562
(2H, m, H-5) 1.46–1.31 (4H, m, H-6,7), 1.10 (3H, t, J¼7.1 Hz, H-
b
);
References and notes
13C NMR (CDCl3):
d
148.0, 148.0, 148.0, 147.9, 135.5, 133.8, 133.7,
1. Ja¨nne, J.; Alhonen, L.; Keina¨nen, T. A.; Pietila¨, M.; Uimari, A.; Pirinen, E.;
Hyvo¨nen, M. T.; Ja¨rvinen, A. J. Cell. Mol. Med. 2005, 9, 865–882.
2. Wallace, H. M.; Fracer, A. V.; Hughes, A. Biochem. J. 2003, 376, 1–14.
3. Gugliucci, A. Clin. Chim. Acta 2004, 344, 23–35.
4. Karigiannis, G.; Papaioannou, D. Eur. J. Org. Chem. 2000, 1841–1863.
5. Casero, R. A., Jr.; Marton, L. J. Nat. Rev. Drug Discov. 2007, 6, 373–389.
6. Casero, R. A., Jr.; Woster, P. M. J. Med. Chem. 2001, 44, 1–26.
7. Boncher, T.; Bi, X.; Varghese, S.; Casero, R. A., Jr.; Woster, P. M. Biochem. Soc.
Trans. 2007, 35, 356–363.
133.7, 133.6, 133.3, 133.1, 133.0 (2C), 132.0, 132.0, 132.0, 131.9,
130.8, 130.8, 130.6, 130.6, 128.8 (2C), 128.4 (2C), 128.2, 124.3,
124.2 (2C), 124.2 (totally 30C–Ph), 52.0 (Ph–CH2–), 42.5 (C-
13.6 (C-
), rest in Table 10; HRMS (ESI-MS): calcd for (MþNa)
C43H40D8N8O16S4Na 1091.2471, found 1091.2467.
a),
b
4.3.67. N-(3-Benzylaminopropyl)-N0-(3-ethylaminopropyl)butane-
1,4-diamine tetrahydrochloride 4e
8. Wallace, H. M.; Niiranen, K. Amino Acids 2007, 33, 261–265.
9. Seiler, N. Pharmacol. Ther. 2005, 107, 99–119.
Prepared as 4a from 14e (927 mg, 0.87 mmol) to give 4e
(284 mg, 70%) as a colourless solid, mp >275 ꢁC decomp. IR (KBr):
10. Bergeron, R. J.; Wiegand, J.; Weimar, W. R.; Snyder, P. S. Pharmacol. Res. 1998, 38,
367–380.
11. Prakash, C.; Shaffer, C. L.; Nedderman, A. Mass Spectrom. Rev. 2007, 26, 340–369.
12. Kostiainen, R.; Kotiaho, T.; Kuuranne, T.; Auriola, S. J. Mass Spectrom. 2003, 38,
357–372.
3050–2389, 1593, 1460, 873, 743, 698 cmꢀ1; 1H NMR (D2O):
d
7.56–
7.47 (5H, m, Ph), 4.28 (2H, s, PhCH2–), 3.24–3.05 (14H, m, H-
,1,3,5,8,10,12), 2.19–2.06 (4H, m, H-2,11), 1.85–1.73 (4H, m, H-6,7),
1.30 (3H, t, J¼7.3 Hz, H- 133.3, 132.7 (2C), 132.7,
); 13C NMR (D2O):
132.2 (2C) (totally 6C–Ph), 54.1 (Ph–CH2–), 45.9 (C- ), 13.3 (C- ),
13. Hakkinen, M. R.; Keina¨nen, T. A.; Vepsa¨la¨inen, J.; Khomutov, A. R.; Alhonen, L.;
a
Janne, J.; Auriola, A. J. Pharm. Biomed. Anal. 2007, 45, 625–634.
14. Hakkinen, M. R.; Keina¨nen, T. A.; Vepsa¨la¨inen, J.; Khomutov, A. R.; Alhonen, L.;
Ja¨nne, J.; Auriola, A. J. Pharm. Biomed. Anal. 2008, 48, 414–421.
15. Stokvis, E.; Rosing, H.; Beijnen, J. H. Rapid Commun. Mass Spectrom. 2005, 19,
401–407.
¨
b
d
a
b
rest in Table 6; HRMS (ESI-MS): calcd for (MþH) C19H37N4 321.3018,
found 321.3003.
16. Cleland, W. W. Arch. Biochem. Biophys. 2005, 433, 2–12.
17. Lee, K. Y.; Na, J. E.; Lee, M. J.; Kim, J. N. Tetrahedron Lett. 2004, 45, 5977–5981.
18. Kuksa, V.; Buchan, R.; Kong Thoo Lin, P. Synthesis 2000, 1189–1207.
19. Gawandi, V.; Fitzpatric, P. F. J. Labelled Compd. Radiopharm. 2007, 50, 666–670.
20. Maruyoshi, K.; Demura, T.; Sagane, T.; Matsumori, N.; Oishi, T.; Murata, M.
Tetrahedron 2004, 60, 5163–5170.
21. Smith, R. G.; Daves, G. D., Jr. Biomed. Mass Spectrom. 1977, 4, 146–151.
22. Jentgens, C.; Bienz, S.; Hesse, M. Helv. Chim. Acta 1997, 80, 1133–1143.
23. Schere, G.; Limbach, H.-H. J. Am. Chem. Soc. 1994, 116, 1230–1239.
24. Van Den Berg, G. A.; Elzinga, H.; Nagel, G. T.; Kingma, A. W.; Muskiet, F. A.
J. Biochim. Biophys. Acta 1984, 802, 175–187.
4.3.68. N-(3-Benzylamino-1,1,2,2-2H4-propyl)-N0-(3-ethylamino-
propyl)butane-1,4-diamine tetrahydrochloride 4f
Prepared as 4a from 14f (2.65 g, 2.48 mmol) to give 4f (790 mg,
68%, 98% d) as a colourless solid, mp >300 ꢁC decomp. IR (KBr):
3050–2311, 1594, 1455, 825, 743, 698 cmꢀ1; 1H NMR (D2O):
d
7.56–
7.47 (5H, m, Ph), 4.29 (2H, s, PhCH2–), 3.24–3.06 (12H, m, H-
,1,5,8,10,12), 2.18–2.06 (2H, m, H-11), 1.86–1.75 (4H, m, H-6,7), 1.31
(3H, t. J¼7.3 Hz, H- 133.3, 132.7 (2C), 132.6,
); 13C NMR (D2O):
132.2 (2C) (totally 6C–Ph), 54.1 (Ph–CH2–), 45.9 (C- ), 13.3 (C- ),
a
´
25. Fodor-Csorba, K.; Galli, G.; Holly, S.; Gacs-Baitz, E. Tetrahedron Lett. 2002, 43,
b
d
3789–3792.
a
b
26. Moser, M.; Hudlicky, T.; Sadechi, S.; Sternin, E. J. Labelled Compd. Radiopharm.
2007, 50, 671–674.
27. Concello´ n, J. M.; Rodrı´guez-Solla, H.; Concello´ n, C. Tetrahedron Lett. 2004, 45,
2129–2131.
28. Whitmore, F. C.; Mosher, H. S.; Adams, R. R.; Taylor, R. B.; Chapin, E. C.; Weisel,
C.; Yanko, W. J. Am. Chem. Soc. 1944, 66, 725–731.
29. Richard, J. P.; Williams, G.; Gao, J. J. Am. Chem. Soc. 1999, 121, 715–726.
30. Iwasaki, M.; Sakka, T.; Ohashi, S.; Matsushita, H. J. Phys. Chem. 1989, 93, 5139–
5143.
rest in Table 6; HRMS (ESI-MS): calcd for (MþH) C19H33N4D4
325.3269, found 325.3260. Isotope distribution: 0% d0–d2, 8% d3,
92% d4.
4.3.69. N-(3-Benzylamino-1,1,2,2-2H4-propyl)-N0-(3-ethylamino-
1,1,2,2-2H4-propyl)butane-1,4-diamine tetrahydrochloride 4g
Prepared as 4a from 14g (1.29 g, 1.21 mmol) to give 4g
(381 mg, 66%, 98% d) as a colourless solid, mp >300 ꢁC decomp.
31. Hidai, Y.; Kan, T.; Fukuyama, T. Chem. Pharm. Bull. 2000, 48, 1570–1576.
32. Jarvinen, A. J.; Cerrade-Gimenez, M.; Grigorenko, N. A.; Khomutov, A. R.;
Vepsa¨la¨inen, J. J.; Sinervirta, R. M.; Keina¨nen, T. A.; Alhonen, L. I.; Ja¨nne, J. E.
J. Med. Chem. 2006, 49, 399–406.
¨
IR (KBr): 3050–2312, 1593, 1452, 822, 744, 698 cmꢀ1
(D2O):
(10H, m, H-
J¼7.3 Hz, H-
132.2 (2C) (totally 6C–Ph), 54.1 (Ph–CH2–), 45.9 (C-
;
1H NMR
7.56–7.46 (5H, m, Ph), 4.29 (2H, s, PhCH2–), 3.24–3.05
,1,5,8,12), 1.85–1.74 (4H, m, H-6,7), 1.30 (3H, t,
); 13C NMR (D2O):
133.3 (Ph), 132.7 (2C), 132.6,
), 13.3 (C- ),
d
33. Grigorenko, N. A.; Khomutov, A. R.; Keina¨nen, T. A.; Ja¨rvinen, A.; Alhonen, L.;
Ja¨nne, J.; Vepsa¨la¨inen, J. Tetrahedron 2007, 63, 2257–2262.
34. Pretsch, E.; Bu¨hlmann, P.; Affolter, C. Structure Determination of Organic Com-
pounds: Tables of Spectral Data 3rd Completely revised and enlarged Engl. ed.;
Springer: Berlin, 2000; p 247.
a
b
d
a
b
rest in Table 6; HRMS (ESI-MS): calcd for (MþH) C19H29N4D8
329.3520, found 329.3518. Isotope distribution: 0% d0–d5, 1% d6,
12% d7, 87% d8.
35. Rouvier, E.; Pastor, R.; Musso, J.; Cambon, A. Org. Magn. Reson. 1974, 6, 640–643.
36. Zhang, H.; Zhang, Y.; Liu, L.; Xu, H.; Wang, Y. Synthesis 2005, 13, 2129–2136.
37. Pearson, D. E.; Jones, W. H.; Cope, A. C. J. Am. Chem. Soc. 1946, 68, 1225–1229.
38. Dawber, J. G.; Massey-Shaw, J. J. Chem. Soc., Perkin Trans. 2 1989, 1249–1254.
39. Compounds 1a, 1h and 1j as free base: Tarbell, D. S.; Shakespeare, N.; Claus,
C. J.; Bunnett, J. F. J. Am. Chem. Soc. 1946, 68, 1217–1219.
40. King, J. A.; McMillan, F. H. J. Am. Chem. Soc. 1946, 68, 1468–1470.
41. Martin, B.; Posse´me´, F.; Le Barbier, C.; Carreaux, F.; Carboni, B.; Seiler, N.;
Moulinoux, J.-P.; Delcros, J.-G. J. Med. Chem. 2001, 44, 3653–3664.
42. Bolkenius, F. N.; Seiler, N. Biol. Chem. Hoppe-Seyler 1989, 370, 525–531.
43. Compound 1e as free base: Surrey, A. J. Am. Chem. Soc. 1949, 71, 3354–3356.
44. Wysocka-Skrzela, B.; Cholody, W. M.; Ledochowski, A. Pol. J. Chem. 1981, 55,
2211–2214.
45. Wagner-Jauregg, T. Helv. Chim. Acta 1961, 44, 1237–1249.
46. Callery, P. S.; Subramayam, B.; Yuan, Z.-M.; Pou, S.; Geelhaar, L. A.; Reynolds,
K. A. Chem.-Biol. Interact. 1992, 85, 15–26.
47. Bergeron, R. J.; Feng, Y.; Weimar, W. R.; McManis, J. S.; Dimova, H.; Porter, C.;
Raisler, B.; Phanstiel, O. J. Med. Chem. 1997, 40, 1475–1494.
48. Carboni, B.; Benalil, A.; Vaultier, M. J. Org. Chem. 1993, 58, 3736–3741.
49. da Silva, E. T.; Fona, F. S.; Lima, E. L. S. J. Braz. Chem. Soc. 2004, 15, 433–436.
50. Olsen, C. A.; Jorgensen, M. R.; Witt, M.; Mellor, I. R.; Usherwood, P. N. R.;
Jaroszewski, J. W.; Franzyk, H. Eur. J. Org. Chem. 2003, 3288–3299.
51. Bergeron, R. J.; McManis, J. S.; Liu, C. Z.; Feng, Y.; Weimar, W. R.; Luchetta, G. R.;
Wu, Q.; Ortiz-Ocasio, J.; Vinson, J. R. T.; Kramer, D.; Porter, C. J. Med. Chem. 1994,
37, 3464–3476.
4.3.70. N-(3-Amino-1,1,2,2-2H4-propyl)-N0-(3-ethylamino-propyl)-
butane-1,4-diamine tetrahydrochloride 3g
Prepared as 1l from 4f (400 mg, 0.85 mmol) to give 3g
(259 mg, 80%, 98% d) as
decomp. IR (KBr): 3000–2388, 1610, 1459, 1353, 1147,
823 cmꢀ1 1H NMR (D2O):
3.22–3.07 (12H, m, H- ,1,3,5,8,12),
2.18–2.06 (2H, m, H-2), 1.86–1.75 (4H, m, H-6,7), 1.30 (3H, t,
); 13C NMR (D2O):
45.9 (C- ), 13.3 (C- ), rest in
a
colourless solid, mp >300 ꢁC
;
d
a
J¼7.3 Hz, H-
b
d
a
b
Table 6; HRMS (ESI-MS): calcd for (MþH) C12H27N4D4
235.2800, found 235.2788. Isotope distribution: 0% d0–d2, 6%
d3, 93% d4.
Acknowledgements
We thank Ms. Maritta Salminkoski, Department of Biosciences,
Laboratory of Chemistry, University of Kuopio, for her help with the
synthesis work. This work was supported by Academy of Finland
(project 124 185) and the Russian Foundation for Basic Research
(project 06-04-49638).
52. Tomasi, S.; Le Roch, M.; Renault, J.; Corbel, J.-C.; Uriac, P.; Carboni, B.; Moncoq,
D.; Martin, B.; Delcros, J.-G. Bioorg. Med. Chem. Lett. 1998, 8, 635–640.
53. Bergeron, R. J.; Wiegand, J.; McManis, J. S.; Weimar, W. R.; Smith, R. E.;
Algee, S. E.; Fannin, T. L.; Slusher, M. A.; Snyder, P. S. J. Med. Chem. 2001, 44,
232–244.
54. Weitl, F. L.; Raymond, K. N. J. Org. Chem. 1981, 46, 5234–5237.