Synthesis and biological evaluation of oxindoles and benzimidazolinones derivatives

Article abstract of DOI:10.1016/j.ejmech.2004.01.001

The synthesis of new oxindoles and benzimidazolinones derivatives bearing a sugar residue on the aromatic nitrogen is described. The presence of the glycoside moiety should enhance the solubility of these heterocyclic compounds and/or improve the interact

Full text of DOI:10.1016/j.ejmech.2004.01.001

European Journal of Medicinal Chemistry 39 (2004) 453–458
www.elsevier.com/locate/ejmech
Short communication
Synthesis and biological evaluation of oxindoles
and benzimidazolinones derivatives
Samir Messaoudi a, Martine Sancelme a, Valérie Polard-Housset b, Bettina Aboab a,
Pascale Moreau a,*, Michelle Prudhomme a
a Laboratoire SEESIB, Université Blaise Pascal, UMR 6504 du CNRS, 24, avenue des Landais, 63177 Aubière cedex, France
b Aventis Pharma S. A., oncology department, biochemistry unit, centre de recherche de Paris, 13, quai Jules Guesde, 94400, Vitry sur Seine, France
Received 4 September 2003; received in revised form 5 January 2004; accepted 15 January 2004
Abstract
The synthesis of new oxindoles and benzimidazolinones derivatives bearing a sugar residue on the aromatic nitrogen is described. The
presence of the glycoside moiety should enhance the solubility of these heterocyclic compounds and/or improve the interaction with the active
site of the biological targets. The inhibitory activities of these new compounds toward five kinases were examined: KDR (VEGFR-2), FGFR-1,
PDGFR-b, EGFR and Tie 2. Furthermore, the antibacterial activities of the prepared compounds were tested against two Gram-positive
bacteria Bacillus cereus and Streptomyces chartreusis, a Gram-negative bacterium Escherichia coli and a yeast Candida albicans.
© 2004 Elsevier SAS. All rights reserved.
Keywords: Oxindoles; Benzimidazolinones; Kinases inhibitors; Antimicrobial agents
1. Introduction
compounds and/or improve the interaction with the active
site of the target enzymes. The inhibitory activities of these
new compounds toward five kinases were examined: KDR
(VEGFR-2), FGFR-1, PDGFR-b, EGFR and Tie 2 (receptor
tyrosine kinase which is activated by its ligand: angiopoietin
1). Since different kinases activities are involved in the
growth of microorganisms and many oxindoles are kinases
inhibitors, these new compounds could exert antibacterial
activities as it was shown previously for several oxindoles
derivatives [5,6]. For these reasons, the antibacterial activi-
ties of the prepared compounds were tested against two
Gram-positive bacteria Bacillus cereus and Streptomyces
chartreusis, a Gram-negative bacterium Escherichia coli and
a yeast Candida albicans.
Growth factors receptors exhibiting a tyrosine kinase ac-
tivity are a group of transmembrane proteins involved in
signal transduction. Their function in many cell types is to
drive a wide variety of cellular functions, including growth,
differentiation and angiogenesis by transducing growth fac-
tor signals from the external environment to intracellular
processes. In malignancies, these pathways are often ex-
ploited by tumor cells to optimize tumor growth and metasta-
sis. Indeed, alterations in receptor tyrosine kinases pathways
have been implicated in oncogenic activation, tumor angio-
genesis and mitogenic stimulation [1]. Thus, receptor ty-
rosine kinases are logical targets for novel anticancer agents
development. Among a large number of small molecule re-
ceptor tyrosine kinases antagonists, several oxindole deriva-
tives (ATP competitive inhibitors) are in phase I–III clinical
development (Fig. 1) [1–4].
2. Chemistry
In this paper, the synthesis of new oxindoles and benzimi-
dazolinones derivatives bearing a sugar residue on the aro-
matic nitrogen are described. The presence of the glycoside
moiety should enhance the solubility of these heterocyclic
1-(b-D-glucopyranosyl)-indolin-2-one 6 was prepared in
five steps via the corresponding indolic intermediate 3 which
was synthesized by the indoline–indole method described by
Mel’nik et al. [7]. Protection of the hydroxyl groups of the
sugar moiety was performed with sodium hydride and benzyl
bromide in the presence of tetrabutylammonium iodide [8].
Protected compound 4 was then brominated with pyridinium
* Corresponding author.
E-mail address: pmoreau@chimsrv1.univ-bpclermont.fr (P. Moreau).
© 2004 Elsevier SAS. All rights reserved.
doi:10.1016/j.ejmech.2004.01.001

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S. Messaoudi et al. / European Journal of Medicinal Chemistry 39 (2004) 453–458
NEt2
O
COOH
N
H
N
H
R
N
H
N
H
O
F
N
O
O
H
N
N
H
H
SU5416 R = H
SU6668
SU11248
VEGFR-2 inhibitor
VEGRF-2, PDGFR-β, FGFR-1 inhibitor
VEGFR-2 and PDGFR-β inhibitor
Phase III clinical trials
Phase II clinical trials
Phase I clinical trials
Fig. 1.
bromide perbromide and oxidized with peracetic acid lead-
ing to 3,3′-dibromo-1-(2,3,4,6-tetra-O-benzyl-b-D-gluco-
pyrannosyl)-indolin-2-one 5 [9,10]. Finally, compound 6
was obtained by hydrogenolysis of the benzyl groups with
concomitant elimination of the bromine atoms (Scheme 1).
Deprotection of the hydroxyl groups of the sugar moiety
to give compound 9 was achieved using N,N-dimethylaniline
and aluminium chloride [13].
To evaluate the influence of modifications on the hetero-
cyclic moiety on the biological activity, especially the pres-
ence of a free NH in a position with respect to the carbonyl
group, compound 12, isoster analogue of 6 with one nitrogen
atom in position 3 was prepared from commercially available
benzimidazolinone 10 (Scheme 3). Coupling with tetra-O-
acetyl-a-bromoglucose was performed with mercuric acetate
in the presence of pyridine [14]. Compound 12 was obtained
from 11 by deprotection of the hydroxyl groups of the sugar
residue using potassium cyanide [15].
Since in the previous described inhibitors of receptor
tyrosine kinases, a conjugated aromatic system is attached in
the 3 position of the oxindoles, compound 9, 3-benzylidenyl
analogue of 6, was prepared in six steps via the indolic
intermediate 4 (Scheme 2). The isatine derivative 7 was
obtained by oxidation of 4 using chromium oxide [10]. Treat-
ment of 7 with benzyltriphenylphosphonium bromide and
butyllithium led to compound 8 as a mixture of Z/E isomers.
By analogy to the results obtained by Tacconi et al. [11] for
the 3-benzylidenyl-indolin-2-one, Z and E vinylic protons
were attributed, respectively, to the signals at 7.53 and
7.86 ppm. The Z/E isomers ratio (25/75) was determined
from the 1H NMR spectrum (CDCl3) on the vinylic protons
signals. These results were confirmed using quantum semi-
empirical calculations (Table 1). The comparison of the Ea
activation energy (energy difference between the reactants
and the transition state) of Z isomer and E isomer for com-
pound 8 showed that E configuration is the major one. The
value of 0.7 Kcal mol–1 of energy difference between Ea(Z)–
Ea(E) indicated that the theoretical Z/E isomers ratio is equal
to the experimental Z/E isomers ratio [12].
3. Results and discussion
3.1. Kinases inhibition
The in vitro kinase inhibitory activities were tested toward
five kinases. Two structurally related split kinase domain
RTKs (PDGFR-b, FGFR-1), endothelial cell specific recep-
tor (KDR, Tie2) and EGFR. Compound 9 did not inhibit the
kinases tested, it slightly activates the kinases. Only EGFR
and Tie 2 were weakly inhibited by compound 6 and 12.
Compared to the references used: staurosporine and SU5614
(an analogue of SU5416 (Fig. 1) with R = Cl), the results of
OH
O
HO
HO
OH
BnBr, NaH
DDQ, dioxane
OH
N
N
Bu4NI, THF
OH
N
N
EtOH, H2O
OBn
OH
H
O
O
O
1
BnO
HO
HO
OBn
OBn
OH
OH
OH
OH
2
3
4
Br
Br
H2/Pd/C
1) PBPB, tBuOH
O
O
N
N
2) CH3COOH, H2O2
MeOH/AcOEt
OBn
OH
O
O
BnO
HO
OBn
OBn
OH
OH
6
5
Scheme 1.

S. Messaoudi et al. / European Journal of Medicinal Chemistry 39 (2004) 453–458
455
O
3.2. Antimicrobial properties
CrO3
CH3CO2H
+
PhCH2PPh3,Br-
O
N
The antimicrobial activities were tested against two
Gram-positive bacteria (Bacillus cereus and Streptomyces
chartreusis), a Gram-negative bacterium (Escherichia coli)
and a yeast (Candida albicans) (Table 2). Compound 6 was
inactive against the microorganisms tested. Compound 9,
analogue of 6 with a benzylidenyl substituent at the 3 posi-
tion was active against B. cereus. The most efficient was
compound 12, an isoster of 6 with one nitrogen atom at the
3 position. This compound inhibits strongly the growth of the
Gram-positive bacteria tested and that of E. coli. Moreover,
compound 12 inhibits the growth of the yeast C. albicans.
In conclusion, a novel series of oxindoles and benzimida-
zolinones derivatives bearing a sugar residue on the aromatic
nitrogen has been synthesized. Only the oxindole 6 and the
benzimidazolinone 12 exhibited weak inhibitory activities
against EGFR and Tie 2 kinases. However, compound 12 was
strongly active against the bacteria tested. The biological
targets of compound 12 remain to be determined. The phar-
maceutical profile of this novel series could be optimized by
structural modifications such as substitutions on the aromatic
moiety and/or on the carbohydrate part. These modifications
are currently in progress in our laboratory.
N
OBn
BuLi, THF
H2O, acetone
OBn
O
O
BnO
BnO
OBn
OBn
OBn
OBn
4
7
CHPh
O
CHPh
AlCl3, PhNMe2
CH2Cl2
O
N
N
OBn
OH
O
O
BnO
HO
OBn
OBn
OH
OH
8
9
E/Z 75:25
Scheme 2.
the inhibitory activities of these new compounds toward
EGFR and Tie 2 were not very significative.
The potencies and selectivities toward various tyrosines
kinases could be highly improved by chemical modifications
on the aromatic moiety [16,17]. These chemical modifica-
tions are currently in progress in our laboratory with the aim
of optimizing the biological activities of compounds 6 and
12.
4. Experimental
Table 1
4.1. Chemistry
Computational results of isomers E/Z ratio for compound 8
E isomer
Z isomer
IR spectra were recorded on a perkin-Elmer 881 spec-
trometer (m in cm–1). NMR spectra were performed on a
Bruker AC 400 (1H: 400 MHz, 13C: 100 MHz) (chemical
shifts d in ppm, the following abbreviations are used: singlet
(s), doublet (d), triplet (t), doubled triplet (dt), doubled dou-
blet (dd), doubled doubled doublet (ddd), multiplet (m),
Ea activational energy (Kcal mol–1
)
26.5
27.2
Ea(Z)–Ea(E) (Kcal mol–1
Theoretical ratio E/Z
Experimental ratio E/Z
)
0.7
75%
75%
25%
25%
The activation energy Ea is the energy difference between the reactants and
the transition state.
H
N
H
N
H
N
i) Hg(OAc) ,pyridine
2
KCN, MeOH
O
O
O
N
N
N
ii) Acetobromoglucose
benzene
H
O
O
OAc
OH
10
AcO
HO
AcO
HO
OAc
OH
11
12
Scheme 3.
Table 2
Antimicrobial activities of compound 6,9,12 against two Gram-positive bacteria (Bacillus cereus and Streptomyces chartreusis), a Gram-negative bacterium
(Escherichia coli) and a yeast (Candida albicans)
Cpd
6
9
B. cereus ATCC 14579
S. chartreusis NRRL 11407
E. coli ATCC 11303
C. albicans IP 444
–
–
–
–
–
+
nd
++
–
12
+++
++
The size of the zones of growth inhibition was 13–16 mm (++++), 10–12 mm (+++), 8–9 (++), 7–8 mm (+), 6–7 mm ( ); sp, inhibition of sporulation.

456
S. Messaoudi et al. / European Journal of Medicinal Chemistry 39 (2004) 453–458
broad signal (br s), tertiary carbons (C tert.), quaternary
carbons (C quat.). Mass spectra (ES) were determined on a
high resolution API Qstar Pulsar i at Organon (Riom,
France). Chromatographic purifications were performed by
flash silica gel Geduran SI 60 (Merck) 0.040–0.063 mm or
Kieselgel 60 (Merck) 0.063–0.200 mm column chromatog-
raphy. For purity tests, TLC were performed on fluorescent
silica gel plates (60 F254 from Merck).
allowed to warm up to room temperature before refluxing for
6 h. Before evaporation, florisil (60/100mesh, Prolabo) was
added to the reaction mixture to give a residue which was
purified by flash column chromatography (eluent Cyclohex-
ane–EtOAc 98:2) to give 4 (550 mg, 0.86 mmol, 78% yield)
as a yellow solid: m.p. 90 °C. IR (KBr) tC=C 1610 cm–1. 1
H
NMR (400 MHz, DMSO-d6): 3.65 (d, 1H, J = 9.5 Hz); 3.75
(m, 2H); 3.81 (d, 1H, J = 9.5 Hz); 3.95 (m, 2H); 4.15 (t, 1H,
J = 9.5 Hz); 4.25 (d, 1H, J = 11.0 Hz); 4.50 (d, 1H,
J = 12.5 Hz); 4.55 (d, 1H, J = 12.0 Hz); 4.65 (d, 1H,
J = 11.0 Hz); 4.85 (d, 1H, J = 10.5 Hz); 4,90 (br s, 2H); 5.82
(d, 1H, J = 9.0 Hz, H1′); 6.65 (d, 1H, J = 3.0 Hz); 6.74 (d, 2H,
J = 7.0 Hz); 7.09–7.39 (m, 20H); 7.65 (d, 1H, J = 8.0 Hz);
7.70 (d, 1H, J = 3.5 Hz); 7.74 (d, 1H, J = 8.0 Hz). 13C NMR
4.1.1. 1-(b-D-glucopyranosyl)-indoline (2)
To a solution of indoline 1 (2 g, 16.8 mmol) in a mixture of
ethanol (120 ml) and water (4 ml) was added D-glucose
(1.41 g, 7.82 mmol). The resulting mixture was refluxed for
24 h. Water was added after 7 and 14 h (1.6 ml each time).
The residue obtained after evaporation was purified by flash
column chromatography (eluent EtOAc–MeOH, from 98:2
to 90:10) to give compound 2 (4.48 g, 16.0 mmol, 95% yield)
as a white solid: m.p. 113 °C. IR (KBr) tCO 1270 cm–1, tC=C
1610 cm–1, tOH 3100–3700 cm–1. 1H NMR (400 MHz,
DMSO-d6): 2.91 (m, 2H); 3.12 (m, 1H); 3.21 (m, 1H); 3.30
(m, 2H); 3.38–3.54 (m, 2H); 3.62 (m, 2H); 4.35 (t, 1H,
J = 5.0 Hz, OH); 4.64 (d, 1H, J = 8.0 Hz, H1′); 4.91 (d, 1H,
J = 5.0 Hz, OH); 5.02 (br s, 2H, OH); 6.59 (t, 2H, J = 8.0 Hz);
6.95 (d, 1H, J = 7.0 Hz); 7.00 (d, 1H, J = 8.0 Hz). 13C NMR
(100 MHz, DMSO-d6): 28.3 (CH2); 45.7 (CH2); 61.5 (C6′);
70.7; 71.4; 78.4; 78.8; 85.7 (C1′, C2′, C3′, C4′, C5′); 108.1;
118.3; 125.0; 127.6 (Ctert.); 130.2; 151.3 (Cquat).
(100 MHz, DMSO-d6): 68.7; 72.3; 73.3; 74.1; 74.6 (C6′
+
CH2 benzyl); 76.3; 77.6; 80.2; 83.9; 84.6 (C1′, C2′, C3′, C4′,
C5′); 102.5; 111.1; 120.0; 120.6; 121.6; 126.5; 127.5–128.2
(Ctert.); 128.5; 135.9; 137.5; 138.1 (2C); 138.5 (Cquat).
4.1.4. 3,3′-dibromo-1-(2,3,4,6-tetra-O-benzyl-b-D-gluco-
pyranosyl)-indolin-2-one (5)
To a solution of indolic compound 4 (400 mg,
0.625 mmol) in t-butanol (6.8 ml) was added pyridinium
bromide perbromide (PBPB) (858 mg, 2.7 mmol). The reac-
tion mixture was stirred at 40 °C for 24 h before evaporation
and addition of a mixture EtOAc–water (1:1). After decanta-
tion, the aqueous phase was extracted twice with EtOAc. The
organic phases were washed with saturated aqueous NaCl
before drying over MgSO4 and concentrated under vacuum
to give a crude residue (512 mg, 0.85 mmol) which was used
directly without further purification for the next step. Acetic
acid (10.8 ml) then H2O2 (1.54 ml) were added. After stirring
for 24 h at room temperature, a mixture of EtOAc–water
(1:1) was added. After decantation, the aqueous layer was
extracted twice with EtOAc. The organic phases were dried
over MgSO4 and concentrated under vacuum to give a resi-
due which was purified by flash chromatography (eluent
Cyclohexane–EtOAc 90:10) to give 5 (355 mg, 0.437 mmol,
70% yield) as colorless solid: m.p. 50 °C. IR (KBr) tC=C
1610 cm–1, tC=O 1750 cm–1. 1H NMR (300 MHz, DMSO-
d6): 3.71 (s, 2H); 3.85 (m, 1H); 3.99 (m, 2H); 4.21 (br s, 1H);
4.40 (m, 1H); 4.54 (m, 3H); 4.64 (d, 1H, J = 11.0 Hz); 4.81
(d, 1H, J = 11.0 Hz); 4.89 (s, 2H); 5.66 (d, 1H, J = 9.0 Hz,
H1′); 7.10 (m, 2H); 7.17 (m, 2H); 7.24 (m, 2H); 7.26–7.40
(m, 17H); 7.75 (d, 1H, J = 8.0 Hz). 13C NMR (100 MHz,
DMSO-d6): 68.3; 72.1; 73.7; 74.2; 74.6 (C6′ + CH2 of benzyl
groups); 76.1; 76.4; 77.3; 80.8; 84.4 (C1′, C2′, C3′, C4′, C5′);
124.6; 126.0; 127.4–128.2; 131.8 (Ctert); 130.1; 137.2; 138.0;
138.2 (2C); 138.4 (Cquat); 169.0 (C=O). The signal corre-
sponding to CBr2 was not visible.
4.1.2. 1-(b-D-glucopyranosyl)-indole (3)
To a solution of 2 (850 mg, 3.02 mmol) in 1,4-dioxane
(170 ml) was added DDQ (823 mg, 3.62 mmol). The mixture
was stirred at room temperature for 12 h. After addition of
saturated aqueous NaHCO3 and extraction with EtOAc, the
organic phases were washed with saturated aqueous NaCl
and dried over MgSO4. The solvent was removed and the
residue purified by flash chromatography (eluent EtOAc–
MeOH, 98:2) to give 3 (622 mg, 2.23 mmol, 74% yield) as a
salmon coloured solid: m.p. 90–95 °C. IR (KBr) tC=C
1610 cm–1, tOH 3080–3700 cm–1; 1H NMR (400 MHz,
DMSO-d6): 3.33 (m, 1H); 3.45 (m, 1H); 3.51 (m, 2H); 3.75
(m, 1H); 3.81 (m, 1H); 4.59 (br s, 1H, OH); 5.15 (d, 1H,
J = 5.0 Hz, OH); 5.25 (d, 2H, J = 5.5 Hz, OH); 5.46 (d, 1H,
J = 9.5 Hz, H1′.); 6.51 (d, 1H, J = 3.5 Hz); 7.09 (t, 1H,
J = 7.0 Hz); 7.18 (dt, 1H, J1 = 7.9 Hz; J2 = 1.5 Hz); 7.56 (d,
1H, J = 3.5 Hz); 7.59 (dd, 1H, J1 = 8.0 Hz; J2 = 1.0 Hz); 7.61
(d, 1H, J = 8.0 Hz). 13C NMR (100 MHz, DMSO-d6): 61.0
(C6′); 69.9; 71.7; 77.6; 79.4; 84.8 (C1′, C2′, C3′, C4′, C5′);
101.6; 110.9; 119.7; 120.3; 121.2; 126.3 (Ctert ); 128.5; 136.2
(Cquat ).
4.1.3. 1-(2,3,4,6-tetra-O-benzyl-b-D-glucopyranosyl)-indole
(4)
4.1.5. 1-(b-D-glucopyranosyl)-indolin-2-one (6)
To a solution of 3 (300 mg, 1.1 mmol) in dry THF (15 ml)
cooled to 0 °C was added NaH (420 mg, 60% dispersion in
mineral oil, 13.1 mmol), Bu4NI (31.2 mg, 0.09 mmol), and
benzyl bromide (1.03 ml, 12.9 mmol). The mixture was
A mixture of 5 (187 mg, 0.59 mmol), MeOH (6 ml),
EtOAc (3 ml) and 10% Pd/C (40 mg) was hydrogenated at
room temperature under 1 bar for 24 h. After filtration over
Celite and washing with MeOH and EtOAc, the filtrate was

S. Messaoudi et al. / European Journal of Medicinal Chemistry 39 (2004) 453–458
457
evaporated and the residue purified by flash chromatography
(eluent EtOAc–MeOH, from 98:2 to 90:10) to give 6
(121 mg, 0.41 mmol, 69% yield) as a white solid: m.p.
70–75 °C. IR (KBr) tC=C 1610 cm–1, tC=O 1705 cm–1, tOH
3040–3680 cm–1; HRMS (ES) [M + Na]+ Calcd. for
C14H17NNaO6, 318.0948; Found: 318.0950. 1H NMR
(400 MHz, DMSO-d6): 3.24–3.36 (m, 3H); 3.47 (m, 1H);
3.60 (s, 2H); 3.75 (dd, 1H, J1 = 11.5 Hz, J2 = 5.5 Hz); 3.87
(m, 1H); 4.62 (t, 1H, J = 5.5 Hz, OH); 5.10 (d, 1H, J = 5.0 Hz,
OH); 5.12 (d, 1H, J = 5.0 Hz, OH); 5.20 (d, 1H, J = 9.0 Hz,
H1′); 5.25 (d, 1H, J = 5.5 Hz, OH); 7.05 (t, 1H, J = 7.5 Hz);
7.17 (d, 1H, J = 8.0 Hz); 7.25 (t, 1H, J = 8.0 Hz); 7.30 (d, 1H,
J = 7.5 Hz). 13C NMR (100 MHz, CD3OD): 37.0 (CH2CO);
63.1 (C6′); 70.7; 71.8; 79.3; 81.4; 83.8 (C1′, C2′, C3′, C4′, C5′);
113.2; 124.1; 125.8; 128.9 (Ctert); 126.3; 143.8 (Cquat); 178,0
(C=O).
rated and the residue was purified by chromatography (eluent
Toluene–THF 98:2) to give a mixture of E/Z 8 (109 mg,
0.147 mmol, 98% yield) as a yellow gum: IR (NaCl) tC=C
1610,1640 cm–1, tC=O 1720 cm–1; 1H NMR (400 MHz,
CDCl3) for the major E isomer: 3.72 (m, 3H); 3.88 (m, 3H);
4.33 (d, 1H, J = 11.0 Hz); 4.49 (d, 1H, J = 12.0 Hz); 4.59 (m,
2H); 4.68 (d, 1H, J = 11.0 Hz); 4.91 (m, 3H); 5.68 (d, 1H,
J = 9.0 Hz, H1′); 6.85 (td, 1H, J1 = 7.5 Hz, J2 = 1.0 Hz); 6.96
(m, 1H); 7.02 (m, 2H); 7.09 (m, 2H); 7.15 (t, 1H, J = 7.5 Hz);
7.22–7.33 (m, 17H); 7.46 (m, 3H); 7.61 (d, 1H, J = 7.6 Hz);
7.64 (d, 1H, J = 6.7 Hz); 7.86 (s, 1H, CHPh). 13C NMR
(100 MHz, DMSO-d6): 69.9; 73.5; 75.1; 75.5; 75.9 (C6′
+
CH2 of the benzyl groups); 77.7; 78.0; 78.8; 81.4; 86.1; (C1′,
C2′, C3′, C4′, C5′); 121.7; 127.4; 135.5; 139.4 (2C); 139.5;
139.8 (2C) (Cquat.); 123.5; 123.6; 128.7–129.5; 130.1; 130.6;
131.3; 133.2; 138.7; 139.6 (Ctert.); 168.4 (C=O).
4.1.6. 1-(2,3,4,6-tetra-O-benzyl-b-D-glucopyranosyl)-indo-
lin-2,3-dione (7)
4.1.8. 3-benzylidenyl-1-b-D-glucopyranosyl-indolin-2-one
(9)
To a solution of 8 (152 mg, 0.2 mmol) in CH2Cl2 (0.5 ml),
dimethylaniline (78 µl, 0.6 mmol) and AlCl3 (109 mg,
0.8 mmol) were added. The reaction mixture was stirred at
room temperature for 21 h. Dimethylaniline and AlCl3 were
added after 1 and 2 h (0.1 ml and 100 mg respectively each
time). HCl 2 M (1.0 ml) was added to the reaction mixture
before extraction with EtOAc. The organic phase was
washed successively with 5% aqueous NaHCO3, saturated
aqueous NaCl and water. After drying over MgSO4, the
solvent was removed and the residue purified by chromatog-
raphy (eluent EtOAc–MeOH, from 2:98 to 10:90) to give a
mixture of E/Z 9 (20 mg, 0.05 mmol, 26% yield) as an
orange–brown amorphous solid: IR (KBr) tC=C 1610 cm–1,
tC=O 1705 cm–1, tOH 3040–3680 cm–1. HRMS (ES) [M +
Na]+ Calcd. for C21H21NNaO6, 406.1261; Found: 406.1259.
1H NMR (400 MHz, DMSO-d6) for the major E isomer: 3.30
(m, 1H); 3.37 (m, 2H); 3.52 (m, 1H); 3.77 (m, 1H); 3.91 (m,
1H); 4.65 (br s, 1H, OH); 5.65 (br s, 1H, OH); 5.73 (br s, 1H,
OH); 5.31 (br s, 1H, OH); 5.32 (d, 1H, J = 8.5 Hz, H1′); 6.93
(t, 1H, J = 8.0, Harom.); 7.25 (d, 1H, J = 7.5 Hz); 7.34 (m, 2H);
7.50–7.64 (m, 3H); 7.75 (m, 2H); 7.80 (s, 1H, CHPh).
Chromium oxide (200 mg, 2.00 mmol) was added slowly
to a solution of indole derivative 4 (100 mg, 0.156 mmol) in
acetone (0.5 ml), acetic acid (1.7 ml, 0.3 mmol) and water
(0.55 ml). The reaction mixture was stirred at room tempera-
ture for 2 h. After addition of water and extraction with
EtOAc, the organic phases were washed with water and
saturated aqueous NaCl until neutral pH. After drying over
MgSO4, the solvent was removed and the residue purified by
flash chromatography (eluent Cyclohexane–EtOAc, from
80:20 to 60:40) to give 7 (63 mg, 0.094 mmol, 62% yield) as
a yellow gum: IR (NaCl) tC=C 1610 cm–1, tC=O 1740 cm–1;
HRMS (ES) [M + Na]+ Calcd. for C42H39NNaO7, 692.2618;
Found: 692.2633; 1H NMR (400 MHz, DMSO-d6): 3.71 (d,
2H, J = 3.0 Hz); 3.80 (m, 1H); 3.89 (dt, 1H, J1 = 9.5 Hz,
J2 = 3.5 Hz); 3.96 (t, 1H, J = 8.5 Hz); 4.11 (br s, 1H); 4.36 (d,
1H, J = 12.0 Hz); 4.50 (d, 2H, J = 3.0 Hz); 4.64 (d, 1H,
J = 11.0 Hz); 4.69 (d, 1H, J = 11.5 Hz); 4.81 (d, 1H,
J = 11.0 Hz); 4.90 (s, 2H); 5.54 (d, 1H, J = 9.0 Hz, H1′); 6.95
(d, 2H, J = 7.0 Hz); 7.04–7.13 (m, 3H); 7.20 (t, 1H,
J = 8.0 Hz); 7.25 (dd, 1H, J1 = 7.5 Hz, J2 = 2.5 Hz); 729–7.40
(m, 15H); 7.55 (d, 1H, J = 7.5 Hz); 7.60 (t, 1H, J = 8.0 Hz).
13C NMR (100 MHz, DMSO-d6): 68.4; 72.2; 73.9; 74.2; 74.7
(C6′ + CH2 of the benzyl groups); 76.3; 76.5; 77.3; 80.0; 84.9
(C1′, C2′, C3′, C4′, C5′); 117.5; 137.6; 138.0; 138.1 (2C);
138.4 (Cquat); 123.7; 124.8; 127.4–128.3; 138.2 (Ctert);
157.4; 182.0 (C=O).
4.1.9. 1-(2,3,4,6-tetra-O-acetyl-b-D-glucopyranosyl)-benzi-
midazolin-2-one (11)
Compound 11 was obtained as a white solid according to
the procedure described by Zinner and Peseke [14]: m.p.
1
248 °C (m.p.litt = 244–246 °C) [14], H NMR (400 MHz,
4.1.7. 3-benzylidenyl-1-(2,3,4,6-tetra-O-benzyl-b-D-gluco-
pyranosyl)-indolin-2-one (8)
DMSO-d6): 1.76; 1.90; 1.97; 1.99 (4s, 12H, CH3 acetate);
4.08 (d, 2H, J = 3.0 Hz); 4.28 (dt, 1H, J1 = 10.0 Hz;
J2 = 3.5 Hz); 5.24 (t, 1H, J = 9.5 Hz); 5.53 (t, 1H, J = 9.5 Hz);
5.57 (t, 1H, J = 9.5 Hz); 5.88 (d, 1H, J = 9.0 Hz, H1′); 6.96 (m,
3H); 7.42 (dd, 1H, J1 = 6.0 Hz; J2 = 2.5 Hz); 11.0 (s, 1H, NH).
A solution of butyllithium (2 M in cyclohexane, 0.11 ml)
was added to a suspension of benzyltriphenylphosphonium
bromide (92 mg, 0.21 mmol) in THF (5 ml). This mixture
was stirred at room temperature for 1 h before addition of a
solution of 7 (100 mg, 0.15 mmol) in THF (5 ml). Saturated
aqueous NaCl was added after stirring at room temperature
for 1 h. The aqueous layer was extracted with EtOAc. The
combined organic phases were dried over MgSO4, evapo-
4.1.10. 1-(b-D-glucopyranosyl)-benzimidazolin-2-one (12)
Potassium cyanide (58.6 mg, 0.9 mmol) was added to a
solution of compound 11 (500 mg, 0.9 mmol) in methanol
(10 ml). The mixture was stirred at room temperature for

458
S. Messaoudi et al. / European Journal of Medicinal Chemistry 39 (2004) 453–458
24 h. Evaporation of the reaction mixture gave a residue
which was purified by flash chromatography (eluent EtOAc–
MeOH, from 95:5 to 90:10) to give 12 (154 mg, 0.50 mmol,
56% yield) as a white solid; m.p. 150 °C; IR (KBr) tC=C
1625 cm–1, tC=O 1700 cm–1, tOH 3000–3600 cm–1. HRMS
(ES) [M + Na]+: Calcd. for C13H16N2O6, 319.0900, Found:
Gram-negative bacterium (E. coli ATCC 11303) and a yeast
(C. albicans 444 from the Pasteur Institute, Paris). The
antimicrobial activity was determined by the conventional
paper disk (Durieux No. 268; 6 mm in diameter) diffusion
method using the following nutrient media: Mueller-Hinton
(Difco) for B. cereus and E. coli, Sabouraud agar (Difco) for
C. albicans and Emerson agar (0.4% beef extract, 0.4%
peptone, 1% dextrose, 0.25% NaCl, 2% agar, pH 7.0) for the
Streptomyces strains. Paper disks impregnated with solutions
in DMSO (300 µg of drug per disk) were placed on Petri
dishes. Growth inhibition was examined after 24 h incuba-
tion at 27 °C.
1
319.0906. H NMR (300 MHz, DMSO-d6): 3.29–3.40 (m,
3H); 3.57 (dt, 1H, J1 = 11.5 Hz; J2 = 6.0 Hz); 3.86 (dd, 1H,
J1 = 11.5 Hz; J2 = 6.0 Hz); 4.01 (m, 1H); 4.69 (t, 1H,
J = 5.5 Hz, OH); 5.19 (d, 1H, J = 5.0 Hz, OH); 5.21 (d, 1H,
J = 5.0 Hz, OH); 5.29 (d, 1H, J = 9.5 Hz, H1′.); 5.31 (d, 1H,
J = 5.5 Hz, OH); 7.11 (m, 3H); 7.29 (dd, 1H, J1 = 8.0 Hz;
J2 = 3.0 Hz); 11.0 (s, 1H, NH). 13C NMR (100 MHz,
CD3OD): 63.1 (C6′); 71.5; 71.8; 79.4; 81.4; 84.7 (C1′, C2′,
C3′, C4′, C5′.); 110.8; 112.2; 122.6; 123.4 (Ctert.); 130.1 (2C)
(Cquat.); 156.8 (C=O).
Acknowledgements
The authors are grateful to Christel Bufferne and
Françoise Lombard (Organon, Riom) for the HRMS (ES)
analyses.
4.2. Computational methods
Quantum semi-empirical calculations SAM1 [18] were
carried out using AMPAC 7.0 [19] program. The FULLCHN
(unpublished results) procedure was used to locate the tran-
sition state. From this transition state geometry, the Intrinsic
Reaction Coordinate [20,21] (IRC) calculation was per-
formed to generate the correct reactants and products states
by following the reaction coordinate in both directions lead-
ing to reactants and products.
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–
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0.1 µM for the other kinases.
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buffer (20 mM MOPS, 10 mM MgCl2, 10 mM MnCl2, 1 mM
DTT, 2.5 mM EGTA, 10 mM b-Glycerophosphate, 1 mM
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started by addition of GST-PLC-c as the substrate (2 µg),
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4.4. Antibiogram tests
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Four strains were tested, two Gram-positive bacteria
(B. cereus ATCC 14579, S. chartreusis NRRL 11407), a