Synthesis and preliminary biological evaluation of new derivatives of the marine alkaloid leucettamine B as kinase inhibitors

Article abstract of DOI:10.1016/j.ejmech.2009.10.009

New derivatives of the marine alkaloid leucettamine B were prepared in five steps with overall yields ranging from 23 to 30%. The key step of our strategy has been the sulfur/nitrogen displacement under solvent-free microwave irradiation of (5Z) 5-benzo[1,3]-dioxo-5-ylmethylene-2-ethylsulfanyl-3,5-dihydroimidazol-4-o ne 3 with a mono-protected ethylenediamine 2. After deprotection of the N-Boc group, the amino derivative of leucettamine B 5 was subjected to reductive amination in two steps with retention of configuration of the double bond, to lead to eight new analogs of leucettamine B. The effect of these compounds on CK1α/β, CDK5/p25, and GSK-3α/β were investigated.

Full text of DOI:10.1016/j.ejmech.2009.10.009

European Journal of Medicinal Chemistry 45 (2010) 805–810
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Short communication
Synthesis and preliminary biological evaluation of new derivatives
of the marine alkaloid leucettamine B as kinase inhibitors
a
a
b
b
a
´
Mansour Debdab , Steven Renault , Olivier Lozach , Laurent Meijer , Ludovic Paquin ,
François Carreaux a, Jean-Pierre Bazureau a
,
*
a
ˆ
Universite´ de Rennes 1, Sciences Chimiques de Rennes, UMR CNRS 6226, Groupe Inge´nierie Chimique & Mole´cules pour le Vivant (ICMV), Bat. 10A,
Campus de Beaulieu, Avenue du Ge´ne´ral Leclerc, CS 74205, 35042 RENNES cedex, France
b Station Biologique de Roscoff, CNRS, Protein Phosphorylation and Human Disease, Place G. Tessier, BP 74, 29682 ROSCOFF cedex, France
a r t i c l e i n f o
a b s t r a c t
Article history:
New derivatives of the marine alkaloid leucettamine B were prepared in five steps with overall yields
ranging from 23 to 30%. The key step of our strategy has been the sulfur/nitrogen displacement
under solvent-free microwave irradiation of (5Z) 5-benzo[1,3]-dioxo-5-ylmethylene-2-ethylsulfanyl-3,5-
dihydroimidazol-4-one 3 with a mono-protected ethylenediamine 2. After deprotection of the N-Boc
group, the amino derivative of leucettamine B 5 was subjected to reductive amination in two steps with
retention of configuration of the double bond, to lead to eight new analogs of leucettamine B. The effect
Received 3 June 2009
Received in revised form
25 September 2009
Accepted 1 October 2009
Available online 12 October 2009
of these compounds on CK1
a
/b
, CDK5/p25, and GSK-3
a
/
b
were investigated.
Keywords:
Marine alkaloid
Ó 2009 Elsevier Masson SAS. All rights reserved.
Leucettamine B
Sulfur/nitrogen displacement
Reductive amination
Microwave irradiation
Solvent-free
Kinase inhibitors
1. Introduction
possible bioactive molecules based on N-functionalized Leucett-
amine B derivatives bearing an arylmethyl group linked to the 2-
amino function via a short aminoalkyl chain (Fig. 2). For this
project, we envisaged to prepare an advanced scaffold on which
we could bring molecular diversity by using simple reactions
such as reductive amination. This intermediate 5 could be built
from (5Z) 5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-ethyl-
sulfanyl-3,5-dihydroimidazol-4-one 3 [6] and an appropriate
symmetric alkyldiamine as precursor of the side chain appended
on the 2-amino imidazolone core. In this paper, we report our
initial results concerning this strategy for the synthesis of a library
of new leucettamine B analogs and their biological evaluation as
protein kinase inhibitors. The 518 protein kinases of the human
kinome constitute a wide family of disease-relevant targets for the
identification and optimization of potential therapeutic agents.
The exploration of marine natural products, in particular those
coming from sponges, continues to make promising contributions
to structurally diverse metabolites that display various types of
biological activity. The 2-aminoimidazolone core is present in an
important number of natural products and constitutes an attractive
scaffold to be used for exploiting chemical diversity and generating
a broad drug-like screening library. Among these, dispacamide,
extracted from Caribbean Agelas sponges, shows antihistaminic
activity [1] (Fig. 1). Polyandrocarpamine A [2], an alkaloid from
a Fijian ascidian, displays selective cytotoxicity towards the SF 268
cell line (central nervous system) with a GI50 value of 65 mM [3].
Leucettamine B isolated in 1993 from the sponge Leucetta micro-
raphis Hæckel (Alcarea class) of the Argulpelu Reef in Palau [4], has
been successfully synthesized by the Bergman’s group [5] but no
biological activity has been reported.
Due to the biological activity associated with the 2-amino
imidazolone moiety, we focused our effort on the synthesis of
2. Chemistry
As illustrated in Scheme 1, the synthesis started with the
preparation of the mono N-Boc protected diamine
commercial ethylenediamine and di-tert-butyldicarbonate.
Complete conversion was observed after 1 h at room temperature
2 from
1
* Corresponding author. Fax: þ33 2 23 23 63 74.
E-mail address: jean-pierre.bazureau@univ-rennes1.fr (J.-P. Bazureau).
0223-5234/$ – see front matter Ó 2009 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2009.10.009

806
M. Debdab et al. / European Journal of Medicinal Chemistry 45 (2010) 805–810
Br
OH
O
Br
O
OR
NH
H N
2
H N
2
H N
2
N
N
N
N
H
O
N
N
N
Me
H
Me
O
O
O
Polyandrocarpamine A (R = Me)
and B (R = H)
Dispacamide
Leucettamine B
Fig. 1. Structure of alkaloids isolated from marine sponges.
in MeOH according to a modified literature method [7], to obtain
the desired product 2 in 65% yield. With the mono N-Boc protected
ethylenediamine 2 in hand, the key step of our strategy is the
guanylation reaction of compound 3, providing access to the
guanidine moiety of 2-amino imidazolone core. In order to be able
to carry out such sulfur/nitrogen displacement in a faster and more
efficient way than the conventional thermal process [8], we
examined the influence of focused microwave irradiations [9]
(SynthewaveÒ 402 reactor [10], Merck Eurolab) with different
reaction times and various powers. After several experiments, we
found that sulfur/nitrogen displacement under microwave irradi-
ation (mu) gave good conversion after 2 h at 105 ꢀC with a stoichi-
ometry of 1/5 of 3/2 to produce the desired product 4 (69%).
For the cleavage of N-Boc group, the use of trifluoroacetic acid in
dichloromethane is often efficient, but in our case, we observed the
formation of some impurities. We thus preferred to use hydro-
chloric acid in an organic solvent [11]. An efficient deprotection of 4
was achieved using 5 equiv of acetyl chloride and 5 equiv of
methanol in MeCN at room temperature [12] for 24 h. After
washing with dry MeOH, the salt 5 was obtained in 98% yield and
the structure was ascertained by high-resolution mass spectrom-
etry (m/z: 288.1218 uma), proton and carbon NMR.
and the structure was confirmed by conventional techniques (1H,
13C NMR and HRMS). Extensive study of 1D and 2D NMR
spectroscopy have confirmed that leucettamine derivatives 8 were
obtained solely as Z isomer and no isomerization about the
exocyclic double bound was observed during our synthesis.
3. Biology
As an initial effort to investigate their in vitro bioactivity,
leucettamine derivatives (5, 7 and 8) were tested against three
protein kinases relevant to Alzheimer’s disease, CK1
kinase 1 ), CDK5 (cyclin-dependent kinase 5)/p25 and GSK-3
(glycogen synthase kinase 3 ) [13–15]. All assays were run in
the presence of 15 M ATP and appropriate protein substrates
(RRKHAAIGpSAYSITA peptide for CK1 [13], histone H1 for CDK5/
p25 [14], GS-1 (YRRAAVPPSPSLSRHSSPHQSpEDEEE) peptide for
GSK-3 [15]). IC50 values were determined from dose-response
a/b (casein
a/b
a/
b
a/b
m
a/b
a/b
curves and are provided in Table 2.
Some leucettamine analogs exhibited a moderate to potent
GSK-3
a
/b
inhibitory activity with IC50 ranging from 0.86 to 38
mM,
and a good selectivity versus CK1a/b
and CDK5/p25 (Table 2).
Interestingly, replacement of an imino group present in compounds
7(a–e) by a less rigid amino function 8(a–e) led to a loss of activity.
Substituents on phenyl group seemed to have a real influence on
the activity of this family if we compare the results between the
different aldimines 7(a–e). However, the N-unsubstituted deriva-
tive 5 preserved a good inhibitory activity and might represent
a scaffold of choice for the discovery of new selective inhibitors of
In order to obtain a large number of compounds suitable for the
biological screening, we privileged the transformation of the salt
5 into N-functionalized derivatives of leucettamine B 8 by reductive
amination in two steps.
We have first examined the reaction of salt 5 with benzaldehyde
6a using various solvents (EtOH, MeOH, MeCN, AcOEt), at different
reaction temperatures and in presence of organic or inorganic bases
(Et3N, pyridine, K2CO3). Among the conditions evaluated, the
reaction of 5 with 2 equiv. of benzaldehyde 6a and Et3N (3 equiv.) in
MeOH (2 ml/mmol.) at 50 ꢀC led to the formation of aldimine 7a in
good yield after 24 h (Table 1). After removal of the solvent in vacuo,
the pure imine 7a was obtained after several washes with a mixture
of EtOH/Et2O (1:1). Having found suitable conditions for this
aldimine synthesis, we screened a range of aromatic aldehydes
6(b–h). As can be seen from the data presented in Table 1, the
leucettamine B derivatives were easily prepared with electron-rich
and electron-poor aromatic aldehydes 6 in yields ranging from 63
to 81%. Finally, the transformation of aldimines 7 into functional-
ized leucettamine B derivatives 8 could be readily accomplished in
good yields (72–97%) using NaBH4 (5 equiv.) in MeOH at 50 ꢀC
during 24 h. According to this approach, the compounds 8(a–h)
were prepared in 5 steps with overall yields ranging from 24 to 31%
GSK-3a/b. Further biological investigations are underway.
4. Conclusion
In summary, we have successfully developed the synthesis of
new derivatives of the marine alkaloid leucettamine B in 5 steps
from commercial ethylenediamine and with good overall yields
(23–30%). One of the key steps of this approach is the sulfur/
nitrogen displacement under microwave irradiation (69%),
demonstrating that microwave is suitable to reduce the reaction
time using solvent-free conditions. Although a limited number of
different and representative substituents of the aromatic group
grafted on the side chain of the 2-amino imidazolone core are
presented here, it is obvious that a much larger diversity can be
achieved. Biological evaluation revealed that this family of
compounds could be considered as new selective inhibitory
O
O
O
O
O
O
H
N
H
N
EtS
Me
N
N
N
NH
H N
2
2
N
H
O
H N
2
R
R
N
N
N
Me
Me
O
O
O
3
5, HCl
Fig. 2. Retrosynthetic strategy towards N-functionalized derivatives of leucettamine B.

M. Debdab et al. / European Journal of Medicinal Chemistry 45 (2010) 805–810
807
O
O
O
O
H
H
N
N
N
N
iii
ii
69%
i
BocHN
NHBoc
H N
2
NH
2
H N
2
H N
2
N
N
65%
O
98%
Me
Me
2
1
O
O
O
4
5, HCl
EtS
Me
N
3
N
iv
O
O
O
R
63-81%
6
O
O
O
H
N
H
N
N
v
N
N
N
H
R
R
N
72-97%
N
Me
Me
8
7
O
O
Scheme 1. Reagents and reaction conditions: (i) (Boc)2O 0.5 equiv., MeOH, 0 ꢀC, 10 min. then 25 ꢀC, 1 h. (ii) 2 5 equiv., mu, 105 ꢀC, 2 h. (iii) MeCOCl 5 equiv., MeOH 5 equiv., MeCN,
25 ꢀC, 24 h. (iv) 6 2 equiv., Et3N 3 equiv., MeOH, 50 ꢀC, 24 h. (v) NaBH4 5 equiv., MeOH, 50 ꢀC, 24 h.
scaffold of GSK-3. This work should enable further biological eval-
uations, analogs synthesis through the synthesis procedure
described here, and studies of the structure–activity relationship
(SAR). These studies are on going in our group.
SynthewaveÒ 402 apparatus (Merck Eurolab, Div. Prolabo, France).
The microwave instrument consists of a continuous focused
microwave power output from 0 to 300 W. All experiments were
performed using stirring option. The target temperature was
reached with a ramp of 5 min and the chosen microwave power
remained constant to keep the mixture at this temperature. The
reaction temperature is monitored using calibrated infrared sensor
and the reaction time include the ramp period. Acetonitrile was
distilled over calcium chloride after standing overnight and stored
over molecular sieves (3 Å). Solvents were evaporated with a BUCHI
rotary evaporator. All reagents were purchased from Acros, Aldrich
Chimie, Fluka France and used without further purification.
The starting (5Z) 5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-
5. Experiment protocols
5.1. General
Melting points were determined on a Kofler melting point
apparatus and were uncorrected. Thin-layer chromatography (TLC)
was accomplished on 0.2-mm precoated plates of silica gel 60
F-254 (Merck) and visualization was made with ultraviolet light
(254 and 312 nm) or with a fluorescence indicator. 1H NMR spectra
were recorded on BRUKER AC 300 P (300 MHz) and 13C NMR
spectra on BRUKER AC 300 P (75 MHz) spectrometer. Chemical
shifts are expressed in parts per million downfield from tetrame-
thylsilane as an internal standard. Data are given in the following
2-ethylsulfanyl-3,5-dihydroimidazol-4-one
according to our previous method [6].
3
was synthesized
5.2. Synthesis of (5Z)-5[(1,3-benzodioxol-5-yl)methylene]-3-
methyl-2-[(2-tert-butylamino)ethylcarbamate]-3,5-
dihydro-4H-imidazol-4-one (4)
order:
d value, multiplicity (s, singlet; d, doublet; t, triplet; q,
quartet; m, multiplet; br, broad), number of protons, coupling
constants J is given in Hertz. The mass spectra (HRMS) were taken
on a VARIAN MAT 311 at an ionizing potential of 70 eV in the Centre
A mixture of (5Z) 5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-
2-ethylsulfanyl-3,5-dihydroimidazol-4-one
3
(1.3 g, 4.5 mmoles)
´
Regional de Mesures Physiques de l’Ouest (CRMPO, Rennes).
and N-Boc protected ethylenediamine 2 (3.6 g, 22.5 mmoles,
Reactions under microwave irradiations were realized in the
5 equiv.) was placed in a cylindrical quartz reactor (Ø ¼ 1.8 cm). The
Table 1
Yield of product (%)a
Reductive amination yield (%)b
Overall yield (%)c
7
8
7a
7b
7c
7d
7e
7f
71
70
72
67
77
81
71
63
8a
8b
8c
8d
8e
8f
88
94
72
85
87
81
90
97
62
66
52
57
67
66
65
61
27
30
23
25
29
29
29
27
7g
7h
8g
8h
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
Cl
O
O N
2
O
6h
6d
6g
6a
Isolated yield.
6b
6c
6e
6f
a
b
c
Yield for 5 –> 8.
Overall yield calculated from commercial ethylenediamine 1.

808
M. Debdab et al. / European Journal of Medicinal Chemistry 45 (2010) 805–810
Table 2
yl)methylene]-3-methyl-2-[(2-aminoethyl)amino]-3,5-dihydro-4H-
imidazol-4-one hydrochloride 5 (0.25 g, 0.77 mmoles) and triethyl-
Kinase inhibitiona values (IC50 in M) for compounds 5, 7 and 8.b
m
amine (0.32 ml, 2.31 mmoles) were mixed in methanol pa (5 ml). To
Compound
CK1
CDK5/p25
GSK-3a/b
this solution was added dropwise
a solution of aldehyde
8a
8b
8c
8d
8e
5
7a
7b
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
> 100
>100
>100
>100
>100
>100
>100
> 10
> 10
> 10
> 10
> 10
1.4
(1.54 mmoles) in 1.5 ml of methanol pa over a period of 30 min. The
mixture was heated at 50 ꢀC for 24 h (monitored by TLC) then cooled
down to room temperature. The solvent was evaporated invacuo and
5 ml of EtOH/Et2O (1:1) were added in the crude reaction mixture.
After vigorous stirring at 25 ꢀC for 30 min, the precipitate that has
formed was filtered off and further dried in a dessicator over CaCl2
under high vacuum (10ꢁ2 Torr) for 6 h. The expected compound 7
was further used without purification and can be stored at 4 ꢀC under
nitrogen.
7.0
0.86
15.0
38.0
7.3
7c
7d
7e
Leucettamine B
15
a
Kinase inhibition experiments were carried out as described previously [15–16].
M.
The compounds, which do not appear in this table, have no activity towards
5.4.1. (5Z)-5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-[(2-(Z)-
benzylideneamino)ethylamino]-3,5-dihydro-4H-imidazol-4-one (7a)
This compound was obtained as yellow solid in 71% yield.
The final ATP-concentration in the test was 15
m
b
these three kinases (>100).
Mp ¼ 200–202 ꢀC. 1H NMR (300 MHz, DMSO)
d: 8.38 (s,1H, CH]N),
8.00 (s,1H, Ar), 7.77–7.81 (m, 3H, Ar, NH), 7.43–7.47 (m, 4H, Ar), 6.91
(d, 1H, J ¼ 8 Hz, Ar), 6.40 (s, 1H, CH]), 6.03 (s, 2H, OCH2O), 3.88 (t,
2H, J ¼ 4.5 Hz, CH2N), 3.70 (t, 2H, J ¼ 4.5 Hz, CH2N), 3.02 (s, 3H,
reactor was then introduced into a SynthewaveÒ 402 Prolabo
microwave reactor. The stirred mixture was irradiated (after a ramp
of 5 min from 20 to 105 ꢀC) at 105 ꢀC (power level: 30%, 90 Watt)
with a reaction time of 2 h. After microwave dielectric heating, the
crude reaction mixture was allowed to cool down at room temper-
ature and Et2O (10 ml) was added in the cylindrical quartz reactor.
The resulting insoluble product 4 was filtered and dried under high
vacuum (10ꢁ2 Torr) for 6 h (yield: 69%).
NCH3). 13C NMR (75 MHz, DMSO)
d: 170.0, 162.8, 158.5, 147.7, 147.2,
139.3, 136.5, 131.2,130.9, 129.1, 128.5, 125.7, 113.5,110.0, 108.8, 101.5,
59.6, 42.6, 26.0. HRMS, m/z: 376.1537 found (calculated for
C21H20N4O3, Mþ. requires: 376.1535).
5.4.2. (5Z)-5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-[{2-(Z)-
(1,3-benzodioxol-5-yl)methyleneaminol}ethylamino]-3,5-dihydro-
4H-imidazol-4-one (7b)
Mp ¼ 108–110 ꢀC. 1H NMR (300 MHz, DMSO)
d: 7.92 (s, 1H, Ar),
7.62 (brs, 1H, NH), 7.42 (d, 1H, J ¼ 8 Hz, Ar), 7.00 (br s, 1H, NH), 6.90
(d, 1H, J ¼ 8 Hz, Ar), 6.37 (s, 1H, CH]), 6.02 (s, 2H, OCH2O), 3.45 (t,
2H, J ¼ 6 Hz, CH2N), 3.24 (t, 2H, J ¼ 6 Hz, CH2N), 3.03 (s, 3H, NCH3),
This compound was obtained as yellow solid in 70% yield.
Mp ¼ 210–212 ꢀC. 1H NMR (300 MHz, DMSO)
d: 8.26 (s, 1H, CH]N),
1.36 (s, 9H, 3 CH3). 13C NMR (75 MHz, DMSO)
d: 169.5, 158.3, 155.7,
8.00 (s, 1H, Ar), 7.77 (br s, 1H, NH), 7.40 (d, 1H, J ¼ 8 Hz, Ar), 7.32 (s,
1H, Ar), 7.22 (d, 1H, J ¼ 8 Hz, Ar), 6.97 (d, 1H, J ¼ 8 Hz, Ar), 6.91 (d,
1H, J ¼ 8 Hz, Ar), 6.40 (s, 1H, CH]), 6.07 (s, 2H, OCH2O), 6.03 (s, 2H,
OCH2O), 3.83 (t, 2H, J ¼ 5 Hz, CH2N), 3.68 (t, 2H, J ¼ 5 Hz, CH2N),
147.2, 146.7, 138.7, 130.3, 125.2, 112.9, 109.6, 108.2, 101.0, 79.1, 41.2,
38.6, 28.2, 25.5. HRMS, m/z: 388.1743 found (calculated for
C19H24N4O5, Mþ. requires: 388.1747).
3.02 (s, 3H, NCH3). 13C NMR (75 MHz, DMSO)
d: 169.5, 161.3, 158.0,
5.3. Synthesis of (5Z) 5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-
2-[(2-aminoethyl)amino]-3,5-dihydro-4H-imidazol-4-one
hydrochloride (5)
149.4,147.8,147.2,146.7,125.2,124.3,112.5,109.5,108.2,108.1,106.0,
101.4, 100.1, 58.8, 42.1, 25.5. HRMS, m/z: 421.1511 found (calculated
for C22H21N4O5, [M þ H]þ requires: 421.1512).
In a 100 ml two-necked round-bottomed flask, provided with
a magnetic stirrer and reflux condenser, (5Z)-5[(1,3-Benzodioxol-5-
yl)methylene]-3-methyl-2-[(2-tert-butylamino)ethylcarbamate]-
3,5-dihydro-4H-imidazol-4-one 4 (1 g, 2.6 mmoles) was mixed in
acetonitrile pa (70 ml). To this slurry was added dropwise succes-
sively methanol (0.63 ml, 15.46 mmoles) and freshly distilled acetyl
chloride (1.1 ml, 15.46 mmoles). After stirring at room temperature
for 24 h and elimination of the solvent in a rotary evaporator under
reduced pressure, the crude residue was submitted to washing with
ethanol (10 ml). The desired product 5 was obtained as yellowish
powder and was further dried under high vacuum (10ꢁ2 Torr) at
25 ꢀC for 9 h (yield: 98%).
5.4.3. (5Z)-5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-[{2-(Z)-
(2,3dihydrofuran-5-yl)methyleneamino}ethylamino]-3,5-dihydro-
4H-imidazol-4-one (7c)
This compound was obtained as yellow solid in 72% yield.
Mp ¼ 200–202 ꢀC. 1H NMR (300 MHz, DMSO)
d: 8.26 (s,1H, CH]N),
8.01 (s, 1H, Ar), 7.77 (br s, 1H, NH), 7.66 (s, 1H, Ar), 7.47 (d, 1H,
J ¼ 8 Hz, Ar), 7.40 (d, 1H, J ¼ 8 Hz, Ar), 6.91 (d, 1H, J ¼ 8 Hz, Ar), 6.80
(d, 1H, J ¼ 8 Hz, Ar), 6.40 (s, 1H, CH]), 6.03 (s, 2H, OCH2O), 4.56 (t,
2H, J ¼ 9 Hz, CH2O), 3.82 (t, 2H, J ¼ 6 Hz, CH2N), 3.67 (t, 2H, J ¼ 6 Hz,
CH2N), 3.16 (t, 2H, J ¼ 9 Hz, CH2), 3.01 (s, 3H, NCH3). 13C NMR
(75 MHz, DMSO) d: 169.5, 162.0, 161.6, 158.0, 147.2, 146.7, 138.8,
130.3, 129.3, 129.0, 128.0, 125.3, 124.3, 112.9, 109.5, 108.7, 108.2,
101.0, 71.5, 59.0, 42.3, 28.5, 25.5. HRMS, m/z: 418.1645 found
(calculated for C23H22N4O4, Mþ. requires: 418.1641).
Mp ¼ 204–206 ꢀC. 1H NMR (300 MHz, DMSO)
d: 8.47 (br s, 3H,
NH2 þ NH), 7.92 (s, 1H, Ar), 7.61 (s, 1H, NH), 7.41 (d, 1H, J ¼ 9 Hz, Ar),
6.98 (d, 1H, J ¼ 9 Hz, Ar), 6.80 (s, 1H, CH]), 6.10 (s, 2H, OCH2O), 3.93
(t, 2H, J ¼ 5 Hz, CH2N), 3.25 (s, 3H, NCH3), 3.20 (t, 2H, J ¼ 5 Hz,
5.4.4. (5Z)-5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-[{2-(Z)-
(2,3-dihydrobenzo[1,4]dioxin-6-yl)methyleneamino}ethylamino]-3,5-
dihydro-4H-imidazol-4-one (7d)
CH2N). 13C NMR (75 MHz, DMSO)
d: 164.5, 156.2, 148.5, 147.6, 126.8,
126.7, 126.5, 117.3, 109.9, 108.6, 101.6, 40.4, 37.9, 26.8. HRMS, m/z:
288.1218 found (calculated for C14H16N4O3, Mþ. requires: 288.1222).
This compound was obtained as yellow solid in 67% yield.
Mp ¼ 202–204 ꢀC. 1H NMR (300 MHz, DMSO)
d:8.23(s,1H, CH]N),
5.4. General procedure for synthesis of compounds 7(a–h)
from product 5 and aldehydes 6(a–h)
8.00 (s, 1H, Ar), 7.76 (br s, 1H, NH), 7.41 (d, 1H, J ¼ 8 Hz, Ar), 7.26 (s,
1H, Ar), 7.24 (d, 1H, J ¼ 8 Hz, Ar), 6.92 (d, 1H, J ¼ 4.1 Hz, Ar), 6.90 (d,
1H, J ¼ 4 Hz, Ar), 6.40 (s, 1H, CH]), 6.03 (s, 2H, OCH2O), 4.26 (m, 4H,
CH2O), 3.82 (t, 2H, J ¼ 6 Hz, CH2N), 3.66 (t, 2H, J ¼ 6 Hz, CH2N), 3.02
In a 10 ml two-necked round-bottomed flask, provided with
a magnetic stirrer and reflux condenser, (5Z) 5-[(1,3-benzodioxo-5-
(s, 3H, NCH3). 13C NMR (75 MHz, DMSO)
d: 169.5, 161.3, 158.0, 147.2,

M. Debdab et al. / European Journal of Medicinal Chemistry 45 (2010) 805–810
809
146.7, 145.6, 143.4, 138.7, 130.3, 129.6, 125.2, 121.5, 117.1, 116.3, 112.9,
109.5, 108.3, 101.0, 64.2, 63.9, 58.9, 42.2, 25.5. HRMS, m/z: 435.1675
found (calculated for C23H23N4O5, [M þ H]þ requires: 435.1669).
30 min. The homogeneous solution was stirred at 50 ꢀC for 24 h
(monitored by TLC). After elimination of solvent in a rotary
evaporator under reduced pressure, deionized water (4 ml) was
added in one portion to the crude residue. The mixture was trans-
ferred to a separating funnel and was extracted with dichloro-
methane (3 ꢂ 5 ml). The combined organic phases were dried over
magnesium sulphate MgSO4, filtered and the solvent was elimi-
nated in vacuo. The desired product 8 was obtained as yellowish
powder and was submitted to purification by recrystallization with
EtOH. Pure compound 8 was dried under high vacuum (10ꢁ2 Torr)
for 6 h at room temperature.
5.4.5. (5Z)-5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-[2-(Z)-
(4-methoxybenzylideneamino)ethylamino]-3,5-dihydro-4H-
imidazol-4-one (7e)
This compound was obtained as yellow solid in 77% yield.
Mp ¼ 194–196 ꢀC. 1H NMR (300 MHz, DMSO)
d:8.30(s, 1H, CH]N),
8.00 (s, 1H, Ar), 7.78 (br s, 1H, NH), 7.71 (d, 2H, J ¼ 9 Hz, Ar), 7.41 (d,
1H, J ¼ 8.5 Hz, Ar), 7.00 (d, 2H, J ¼ 9 Hz, Ar), 6.92 (d, 1H, J ¼ 8.5 Hz,
Ar), 6.38 (s, 1H, CH]), 6.03 (s, 2H, OCH2O), 3.84 (t, 2H, J ¼ 7 Hz,
CH2N), 3.80 (s, 3H, OCH3), 3.67 (t, 2H, J ¼ 7 Hz, CH2N), 3.02 (s, 3H,
5.5.1. (5Z)-5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-[2-
(benzylamino)ethylamino]-3,5-dihydro-4H-imidazol-4-one (8a)
This compound was obtained as yellow solid in 88% yield.
NCH3). 13C NMR (75 MHz, DMSO)
d: 169.5, 161.5, 161.2, 158.0, 147.2,
146.7, 138.8, 130.3, 129.6, 128.8, 125.2, 113.9, 112.9, 109.5, 108.3,
101.0, 59.0, 55.2, 42.2, 25.5. HRMS, m/z: 406.1667 found (calculated
for C22H22N4O4, Mþ. requires: 406.1641).
Mp ¼ 162–164 ꢀC. 1H NMR (300 MHz, CDCl3)
d: 7.97 (s, 1H, Ar), 7.31
(m, 6H, Ar), 6.81 (d, 1H, J ¼ 8 Hz, Ar), 6.65 (s, 1H, CH]), 5.98 (s, 2H,
OCH2O), 5.56 (br s, 2H, NH), 3.84 (s, 2H, CH2N), 3.65 (t, 2H, J ¼ 5 Hz,
CH2N), 3.12 (s, 3H, NCH3), 2.87 (t, 2H, J ¼ 5 Hz, CH2N). 13C NMR
5.4.6. (5Z)-5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-[2-(Z)-
(4-nitrobenzylideneamino)ethylamino]-3,5-dihydro-4H-imidazol-
4-one (7f)
(75 MHz, CDCl3) d: 170.2,157.2,147.7,147.5,139.9,138.1,130.2,128.6,
128.1, 127.3, 126.0, 116.6, 110.2, 108.3, 101.1, 53.3, 47.4, 41.0, 25.1.
HRMS, m/z: 378.1700 found (calculated for C21H22N4O3, Mþ.
requires: 378.1692).
This compound was obtained as red solid in 81% yield.
Mp ¼ 206–208 ꢀC. 1H NMR (300 MHz, CDCl3)
d:8.46(s, 1H, CH]N),
8.26 (d, 2H, J ¼ 8.5 Hz, Ar), 8.00 (s, 1H, Ar), 7.90 (d, 2H, J ¼ 8.5 Hz,
Ar), 7.33 (d, 1H, J ¼ 8 Hz, Ar), 6.82 (d, 1H, J ¼ 8 Hz, Ar), 6.68 (s, 1H,
CH]), 6.00 (s, 1H, OCH2O), 5.18 (br s, 1H, NH), 3.98 (m, 4H, CH2N),
5.5.2. (5Z)-5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-[2-{(1,3-
benzodioxol-5-yl)methylamino}ethylamino]-3,5-dihydro-4H-
imidazol-4-one (8b)
3.13 (s, 3H, NCH3). 13C NMR (75 MHz, CDCl3)
d: 170.1, 160.8, 157.0,
149.2, 147.7, 141.0, 137.8, 130.1, 128.9, 128.8, 126.3, 123.9, 117.5, 110.3,
108.4, 101.1, 60.0, 42.5, 25.1. HRMS, m/z: 421.1351 found (calculated
for C21H19N5O5, Mþ. requires: 421.1386).
This compound was obtained as yellow solid in 94% yield.
Mp ¼ 218–220 ꢀC. 1H NMR (300 MHz, DMSO)
d: 7.97 (s, 1H, Ar), 7.40
(s, 1H, Ar), 6.90 (d, 2H, J ¼ 8 Hz, Ar), 6.76 (d, 2H, J ¼ 8 Hz, Ar), 6.40 (s,
1H, CH]), 6.01 (s, 2H, OCH2O), 5.93 (s, 2H, OCH2O), 5.57 (br s, 2H,
NH), 3.65 (s, 2H, CH2N), 3.51 (t, 2H, J ¼ 5 Hz, CH2N), 3.03 (s, 3H,
5.4.7. (5Z)-5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-[2-(Z)-
(4-chlorobenzylideneamino)ethylamino]-3,5-dihydro-4H-imidazol-
4-one (7g)
NCH3), 2.75 (t, 2H, J ¼ 5 Hz, CH2N). 13C NMR (75 MHz, DMSO)
d:
159.6, 158.1, 147.2, 147.1, 146.6, 145.7, 138.8, 134.9, 130.4, 125.1, 120.8,
112.6, 109.5, 108.2, 112.6, 109.5, 108.3, 108.2, 107.7, 101.0, 100.6, 52.2,
47.3, 41.2, 25.4. HRMS, m/z: 423.1662 found (calculated for
C22H23N4O5, [M þ H]þ requires: 423.1669).
This compound was obtained as yellow solid in 71% yield.
Mp ¼ 192–194 ꢀC. 1H NMR (300 MHz, DMSO)
d:8.40(s, 1H, CH]N),
8.00 (s, 1H, Ar), 7.78 (m, 3H, NH, Ar), 7.50 (d, 2H, J ¼ 8 Hz, ArH), 7.40
(d, 1H, J ¼ 8 Hz, Ar), 6.91 (d; 1H, J ¼ 8 Hz, Ar), 6.40 (s, 1H, CH]), 6.03
(s, 2H, OCH2O), 3.87 (t, 2H, J ¼ 6 Hz, CH2N), 3.70 (t, 2H, J ¼ 6 Hz,
5.5.3. (5Z)-5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-[2-
{(2,3-dihydrobezofuran-5-yl)methylamino}ethylamino]-3,5-
dihydro-4H-imidazol-4-one (8c)
CH2N), 3.02 (s, 3H, NCH3). 13C NMR (75 MHz, DMSO)
d: 170.0, 161.6,
158.7, 147.7, 147.2, 139.2, 135.7, 135.3, 130.8, 130.1, 129.2, 125.7, 113.5,
110.0, 108.8, 101.5, 59.6, 42.5, 26.0. HRMS, m/z: 410.1183 found
(calculated for C21H19N4O3Cl, Mþ. requires: 410.1146).
This compound was obtained as yellow solid in 72% yield.
Mp ¼ 188–190 ꢀC. 1H NMR (300 MHz, CDCl3)
d: 7.97 (s, 1H, Ar), 7.33
(d, 1H, J ¼ 8 Hz, Ar), 7.12 (s, 1H, Ar), 7.03 (d, 1H, J ¼ 8 Hz, Ar), 6.80 (d,
1H, J ¼ 8 Hz, Ar), 6.73 (d, 1H, J ¼ 8 Hz, Ar), 6.63 (s, 1H, CH]), 5.96 (s,
2H, OCH2O), 5.47 (br s, 2H, NH), 4.55 (t, 2H, J ¼ 9 Hz, CH2O), 3.73 (s,
2H, CH2N), 3.62 (t, 2H, J ¼ 4.5 Hz, CH2N), 3.17 (t, 2H, J ¼ 9 Hz, CH2),
3.10 (s, 3H, NCH3), 2.93 (t, 2H, J ¼ 4.5 Hz, CH2N). 13C NMR (75 MHz,
5.4.8. (5Z)-5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-[2-(Z)-
(3,4,5-trimethoxybenzylideneamino)ethylamino]-3,5-dihydro-4H-
imidazol-4-one (7h)
This compound was obtained as yellow solid in 63% yield.
Mp ¼ 198–200 ꢀC. 1H NMR (300 MHz, DMSO)
d
: 8.28 (s, 1H, CH]N),
CDCl3) d: 170.2, 159.4, 157.2, 147.7, 147.5, 138.1, 132.0, 130.2, 128.0,
8.00 (s, 1H, Ar), 7.80 (br s, 1H, NH), 7.41 (d, 1H, J ¼ 8 Hz, Ar), 7.08 (s,
2H, Ar), 6.91 (d, 1H, J ¼ 8 Hz, Ar), 6.40 (s, 1H, CH]), 6.02 (s, 2H,
OCH2O), 3.87 (t, 2H, J ¼ 6 Hz, CH2N), 3.80 (s, 6H, OCH3), 3.70 (m, 5H,
127.3, 126.0, 124.9, 116.8, 110.3, 109.1, 108.3, 101.1, 71.3, 53.0, 47.2,
40.9, 29.7, 25.1. HRMS, m/z: 420.1825 found (calculated for
C23H24N4O4, Mþ. requires: 420.1798).
CH2N, OCH3), 3.02 (s, 3H, NCH3). 13C NMR (75 MHz, DMSO)
d: 170.0,
162.3, 158.5, 153.5, 147.7, 147.2, 140.0, 139.3, 132.0, 130.8, 125.7, 113.4,
110.0, 108.8, 105.6, 101.5, 60.5, 59.5, 56.3, 42.5, 26.0. HRMS, m/z:
466.1823 found (calculated for C24H26N4O6, Mþ. requires: 466.1852).
5.5.4. (5Z)-5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-[2-
{(1,4-benzodioxin-6-yl)methylamino}ethylamino]-3,5-dihydro-4H-
imidazol-4-one (8d)
This compound was obtained as yellow solid in 85% yield.
5.5. General procedure for preparation of compounds 8(a–h)
by reduction of 7(a–h)
Mp ¼ 186–188 ꢀC. 1H NMR (300 MHz, CDCl3)
d: 8.00 (s, 1H, Ar), 7.34
(d, 1H, J ¼ 8 Hz, Ar), 6.80 (m, 4H, Ar), 6.64 (s, 1H, CH]), 5.97 (s, 2H,
OCH2O), 5.50 (br s, 2H, NH), 4.24 (m, 4H, CH2O), 3.71 (s, 2H, CH2N),
3.60 (t, 2H, J ¼ 5 Hz, CH2N), 3.11 (s, 3H, NCH3), 2.92 (t, 2H, J ¼ 5 Hz,
In a 10 ml two-necked round-bottomed flask, provided with
a magnetic stirrer and reflux condenser, compound 7 (0.5 mmoles)
was dissolved in methanol pa (3 ml) under vigorous stirring. To this
solution was added dropwise a solution of sodium borohydride
NaBH4 (0.1 g, 2.5 mmoles) in 2 ml of methanol pa over a period of
CH2N). 13C NMR (75 MHz, CDCl3)
d: 170.3, 157.2, 147.7, 147.5, 143.5,
142.7, 138.2, 133.4, 130.3, 126.0, 121.0, 117.2, 116.8, 110.3, 108.3, 101.1,
64.4, 64.3, 52.7, 47.1, 40.9, 25.1. HRMS, m/z: 436.1790 found
(calculated for C23H24N4O5, Mþ. requires: 436.1747).

810
M. Debdab et al. / European Journal of Medicinal Chemistry 45 (2010) 805–810
5.5.5. (5Z)-5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-[2-(4-
methoxybenzylamino)ethylamino]-3,5-dihydro-4H-imidazol-4-
one (8e)
HRMS, m/z: 468.2018 found (calculated for C24H28N4O6, Mþ.
requires: 468.2019).
This compound was obtained as yellow solid in 87% yield.
5.6. Biology
Mp ¼ 184–186 ꢀC. 1H NMR (300 MHz, CDCl3)
d: 8.00 (s, 1H, Ar), 7.35
(d, 1H, J ¼ 8 Hz, Ar), 7.24 (d, 2H, J ¼ 8 Hz, Ar), 6.88 (d, 2H, J ¼ 8 Hz,
Ar), 6.82 (d, 1H, J ¼ 8 Hz, Ar), 6.66 (s, 1H, CH]), 6.00 (s, 2H, OCH2O),
5.47 (br s, 2H, NH), 3.81 (s, 3H, OCH3), 3.78 (s, 2H, CH2N), 3.63 (t, 2H,
J ¼ 5.5 Hz, CH2N), 3.12 (s, 3H, NCH3), 2.96 (t, 2H, J ¼ 5.5 Hz, CH2N).
Kinase activities were assayed according to the methodology
[16] developed by the Cell Cycle Group of the Station Biologique,
CNRS, Roscoff, France.
13C NMR (75 MHz, CDCl3)
d: 170.3, 158.8, 157.2, 147.7, 147.5, 138.2,
132.0, 130.2, 129.3, 126.0, 116.8, 113.9, 110.3, 108.3, 101.1, 55.3, 52.7,
47.2, 40.9, 25.1. HRMS, m/z: 408.1762 found (calculated for
C22H24N4O4, Mþ. requires: 408.1798).
Acknowledgements
`
´
We thank the ‘‘Ministere de l’Enseignement Superieur et de la
´
´
´
Recherche Scientifique de la Republique Algerienne Democratique
5.5.6. (5Z)-5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-[2-(4-
nitrobenzylamino)ethylamino]-3,5-dihydro-4H-imidazol-4-one (8f)
This compound was obtained as yellow solid in 81% yield.
´
et Populaire (Cooperation et Echanges Inter-universitaires Franco-
´
Algerien CMEP)’’ for M.D. Financial support for this program carried
´
ˆ
out under the ‘‘Canceropole Grand Ouest/Institut National du
Cancer’’, contracts PRIR 04-8390 et ACI 04-2254 are gratefully
acknowledged. We also thank Merck Eurolab Prolabo (Fr.) for
providing the SynthewaveÒ 402 apparatus. LM’s funding includes
CRITT Sante Bretagne, Fondation Jerome Lejeune and Fondation
France Alzheimer (Finiste`re, France).
Mp ¼ 150–152 ꢀC. 1H NMR (300 MHz, DMSO)
d: 8.14 (d, 2H,
J ¼ 8.6 Hz, Ar), 7.96 (s, 1H, Ar), 7.62 (d, 2H, J ¼ 8.6 Hz, Ar), 7.37 (d, 1H,
J ¼ 8 Hz, Ar), 6.90 (d, 1H, J ¼ 8 Hz, Ar), 6.35 (s, 1H, CH]), 6.03 (s, 2H,
OCH2O), 5.50 (br s, 2H, NH), 3.90 (s, 2H, CH2N), 3.53 (t, 2H,
J ¼ 5.5 Hz, CH2N), 3.04 (s, 3H, NCH3), 2.80 (t, 2H, J ¼ 5.5 Hz, CH2N).
´
´ ˆ
13C NMR (75 MHz, DMSO)
d: 169.5, 158.1, 149.5, 147.1, 146.6, 146.2,
138.8, 130.4, 128.7, 125.1, 123.1, 112.6, 109.4, 108.2, 101.0, 51.7, 47.5,
41.1, 25.5. HRMS, m/z: 245.0802 found (calculated for C12H11N3O3,
[M–C9H10N2O2]þ requires: 245.0800).
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[8] The guanylation step can be performed under thermal process but requires
a large excess of amine (solvent) and a long reaction time (>24 h) which
complicates the purification of the final product.
5.5.7. (5Z)-5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-[2-(4-
chlorobenzylamino)ethylamino]-3,5-dihydro-4H-imidazol-4-one (8g)
This compound was obtained as yellow solid in 90% yield.
Mp ¼ 188–190 ꢀC. 1H NMR (300 MHz, DMSO)
d: 8.00 (s,1H, Ar), 7.60
(br s, 2H, NH), 7.40 (d, 1H, J ¼ 7.5 Hz, Ar), 7.34 (m, 4H, Ar), 6.90 (d,
1H, J ¼ 7.5 Hz, Ar), 6.37 (s,1H, CH]), 6.03 (s, 2H, OCH2O), 3.74 (s, 2H,
CH2N), 3.52 (t, 2H, J ¼ 6 Hz, CH2N), 3.04 (s, 3H, NCH3), 2.77 (t, 2H,
J ¼ 6 Hz, CH2N). 13C NMR (75 MHz, DMSO)
d: 170.0, 158.6, 147.7,
147.1, 140.5, 139.3, 131.4, 130.9, 130.1, 128.4, 125.6, 113.1, 110.0, 108.7,
101.5, 52.2, 48.0, 41.6, 26.0. HRMS, m/z: 412.1302 found (calculated
for C21H21N4O3Cl, Mþ. requires: 412.1302).
[9] J.P. Bazureau, F. Mongin, J. Hamelin, F. Texier-Boullet, Microwave in hetero-
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ed. Wiley-VCH, Weinheim, Germany, 2006, pp. 426–523 Chapter 10.
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5.5.8. (5Z)-5-[(1,3-benzodioxo-5-yl)methylene]-3-methyl-2-[2-
(3,4,5-trimethoxybenzylamino)ethylamino]-3,5-dihydro-4H-
imidazol-4-one (8h)
This compound was obtained as yellow solid in 97% yield.
Mp ¼ 184–186 ꢀC. 1H NMR (300 MHz, DMSO)
d: 8.03 (s, 1H, Ar), 7.67
(br s, 2H, NH), 7.44 (d,1H, J ¼ 8 Hz, Ar), 6.93 (d,1H, J ¼ 8 Hz, Ar), 6.67
(s, 2H, Ar), 6.40 (s, 1H, CH]), 6.05 (s, 2H, OCH2O), 3.74 (m, 8H,
CH2N, OCH3), 3.64 (s, 3H, OCH3), 3.60 (t, 2H, J ¼ 6 Hz, CH2N), 3.10 (s,
3H, NCH3), 2.83 (t, 2H, J ¼ 6 Hz, CH2N). 13C NMR (75 MHz, DMSO)
d:
[16] P. Polychronopoulos, P. Magiatis, A.L. Skaltsounis, V. Myrianthopoulos,
E. Mikros, A. Tarricone, A. Musacchio, S.M. Roe, L. Pearl, M. Leost, P. Greengard,
L. Meijer, J. Med. Chem. 47 (2004) 935–946.
169.6, 158.2, 152.6, 147.2, 146.6, 138.9, 136.7, 135.9, 130.4, 125.1,
112.5, 109.4, 108.2, 104.8, 101.9, 59.8, 55.6, 52.6, 47.6, 41.0, 25.4.