Design, synthesis and preliminary biological evaluation of a focused combinatorial library of stereodiverse carbohydrate-scaffold-based peptidomimetics

Article abstract of DOI:10.1016/S0968-0896(00)00256-X

A focused combinatorial library of 126 mimetics of the RGD sequence based on sugar scaffolds have been rationally constructed using molecular modeling, with a particular emphasis on the stereodiversity of the library. A liquid phase, mix and divide synthesis was used, active compounds being identified by using orthogonal libraries and recursive deconvolution strategies.

Full text of DOI:10.1016/S0968-0896(00)00256-X

Bioorganic & Medicinal Chemistry 9 (2001) 511±523
Design, Synthesis and Preliminary Biological Evaluation of
a Focused Combinatorial Library of Stereodiverse
Carbohydrate-Sca€old-Based Peptidomimetics
Nicolas Moitessier,a Sylvie Dufour,c Francoise Chretien,a Jean Paul Thiery,c
Bernard Maigretb and Yves Chapleura,*
a
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Â
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Groupe SUCRES, Unite Mixte 7565 CNRS-Universite Henri Poincare-Nancy 1,
BP 239, F-54506 Nancy-Vandoeuvre, France
Â
b
  Â
Groupe de Biochimie Theorique, Unite Mixte 7565 CNRS-Universite Henri Poincare-Nancy 1, BP 239,
F-54506 Nancy-Vandoeuvre, France
Unite Mixte 144, CNRS-Institut Curie, 24 Rue d'Ulm, F-75000 Paris, France
c
Â
Received 17 July 2000; accepted 4 October 2000
AbstractÐA focused combinatorial library of 126 mimetics of the RGD sequence based on sugar sca€olds have been rationally
constructed using molecular modeling, with a particular emphasis on the stereodiversity of the library. A liquid phase, mix and
divide synthesis was used, active compounds being identi®ed by using orthogonal libraries and recursive deconvolution strategies.
# 2001 Elsevier Science Ltd. All rights reserved.
Introduction
short time with great chances of success. We tried to
make this idea real by studying a de®ned biological
problem: the inhibition of integrin-mediated adhesion, the
dysfunction of which is often related to cancer metastasis,
angiogenesis and osteoporosis.2 Among others, the most
biologically relevant aIIbb3 and avb3 integrins recognize the
short Arg-Gly-Asp (RGD) peptidic sequence exhibited by
their proteic ligands, ®bronectin, vitronectin and other
proteins of the extracellular matrix.3 A major concern
of the adhesion inhibition relies on the selectivity of a
protein vis-a-vis of a particular integrin. This selectivity is
thought to be related to a bioactive conformation of the
RGD sequence which could be speci®c for its receptor.4
The design of soluble RGD mimetics has been largely
investigated for the inhibition of the ®brinogen-aIIbb3
binding mediated platelet aggregation.5 Only a few selec-
tive avb3 integrin antagonists have been found by
screening non-peptide molecules.6 Some peptide analo-
gues have been proposed7 and a new class of stereo-
isomeric peptidic antagonists with high avb3 selectivity
has been reported.8
Combinatorial chemistry considerably changed the way
of ®nding new molecules and also the way one is think-
ing chemistry.1 Most drug research is now concentrated
on the ®nding of lead compounds by high throughput
screening of large combinatorial libraries of chemically
diverse compounds on a high number of pharmacologi-
cal targets. As soon as a lead compound is found, a more
restricted library is constructed by playing around the
original hit compound structure to provide more active
compounds. All this process seems to be contradictory to
the rational drug design in fast development in the eighties
which led, with the help of some knowledge of the biolo-
gical target and of molecular modeling, to the design and
synthesis of only a few candidate molecules.
Between these two approaches, apparently far from each
other, there should be a space for an alternative approach,
faster than the latter and less costly than the former. It
appeared obvious that a rational library design, which
may be computer-aided, would lead directly to a focused
library able to a€ord a few lead compounds in a very
In this work, new selective RGD mimetics based on
chiral sca€olds using several original features including
computer-aided design of the library and a solution
phase combinatorial approach were devised by putting
emphasis on stereodiversity.
*Corresponding author. Tel.: +33-383-91-23-55; fax: +33-383-91-24-
79; e-mail: Yves.Chapleur@meseb.uhp-nancy.fr
0968-0896/01/$ - see front matter # 2001 Elsevier Science Ltd. All rights reserved.
PII: S0968-0896(00)00256-X

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N. Moitessier et al. / Bioorg. Med. Chem. 9 (2001) 511±523
Results
Because the conformation of the RGD sequence of a
ligand is believed important in the selective recognition by
a given integrin, such conformations were tentatively
de®ned by calculations. This approach proved quite useful
in the design9 and synthesis of some aIIbb3 antagonists.10
Taking advantage of previously reported antagonists of
the avb3 integrin receptor,8d a theoretical model of the
possible RGD conformations recognized by the avb3
integrin was built. Furthermore, we decided to construct
stereodiverse libraries, i.e. containing compounds bearing
identical chemical groups but distinct by their respective
shapes. To construct such a library, easily accessible scaf-
folds having di€erent chemical functions suitable for
grafting the required amino acid side chain mimics were
needed. At ®rst glance, mimicking the RGD sequence
can be achieved by using a carboxylic or sulfonic acid
residue for aspartic acid and a basic site as a surrogate
of the guanidine of the arginine residue, limiting the
chemical diversity of the libraries. However, these
chemical groups should have di€erent orientations in
the space, so that each compound would mimic a given
conformation of the native RGD peptidic chain.
Given these requirements, sugar sca€olds seemed excel-
lent candidates owing to the large number of chemical
functions, i.e. hydroxyl groups with the di€erent absolute
con®gurations as existing in carbohydrates. Successful
uses of sugar sca€olds for peptidomimetics construction
can be found in the literature.10,11 Although combina-
torial approaches have been implemented in carbohydrate
chemistry,12,13 to the best of our knowledge, the con-
struction of peptidomimetic libraries on carbohydrate
sca€olds is unprecedented.14
Figure 1. Selective inhibitors of avb3 integrin.
Theoritical model
These studies began with the construction of a theore-
tical model of the RGD sequence conformation which
should selectively bind to the avb3 integrin. This would
complement our previously described aIIbb3 antagonists
model.9 Three highly selective, already existing, antago-
nists shown in Figure 1 were modeled, of which two
arose from peptidomimetics proposed by Merck,15
SmithKline-Beecham16 and the third was a rigidi®ed
cyclic pentapeptide cyclo-RGDfV proposed by Kes-
sler4b,8 (Fig. 1). Modeling of these three molecules was
performed in two steps.9b The ®rst, using simulated
annealing, yielded low-energy conformers which were
visually compared and gathered in di€erent families.
Representative conformations of each of these families
were used for subsequent molecular dynamics in a water
box to explore the accessible energetic hypersurface.
Finally the existing conformers appearing in the course of
the dynamics calculations for the three antagonists were
superimposed and led to a model of the pharmacophore in
which the carboxylic residue had a restricted mobility
whereas the basic function, guanidine or a surrogate,
had a higher mobility and can occupy a wider space.17 The
resulting model showed an amine±carboxylated distance
ranging from 7 to 13 A (Fig. 2). It is interesting to note
that this distance is of the same order of magnitude in the
Figure 2. Theoritical model obtained by molecular dynamics: circles
represent the possible orientation of the carboxylic acid (right) and the
amine groups (left) in the three antagonists in Figure 1.
present model and in our already proposed model for
aIIbb3-selective antagonists.9 However, it was clear from
the latter that these antagonists should need an additional
carboxylate or a surrogate. We suggested that these
antagonists might mimic the dodecapeptide chain of ®bri-
nogen, the aIIbb3-ligand, rather than the RGD sequence.
Libraries construction
From theoretical models it was possible to devise new
molecules based on a sugar sca€old. The extended

N. Moitessier et al. / Bioorg. Med. Chem. 9 (2001) 511±523
513
structure of the pharmacophore led to choosing a car-
bohydrate having all hydroxyl groups equatorially
oriented. In addition, the poorly de®ned bioactive con-
formation of the arginine side chain led us to probe the
surrounding space by using the stereodiversity concept.
Thus the use of both anomers would allow a large
exploration of the space. One or several carboxylic acid
residues should then be introduced on the remaining
hydroxyl groups via O-alkylation. The use of di€erent
anchoring points on the sca€old could modify the
amine±carboxylic acid distance, thus allowing the two
groups to occupy di€erent spatial arrangements and
introducing the stereodiversity. It was then checked by
molecular mechanics calculation±geometry optimization
that low-energy conformers of some representatives of
these structures can more or less adopt conformations
in agreement with the theoretical model. At that point a
more precise ®tting was not required owing to the
planned combinatorial approach. Accordingly d-xylose
was chosen as an appropriate sca€old having three equa-
torial hydroxyl groups with the same type of reactivity.10
A synthetic strategy was then de®ned which made use of
solution phase chemistry and a ``mix and divide'' strategy
to decrease the number of reactions to perform. Thus a
deconvolution strategy was needed to allow the identi®-
cation of the most active compound of the library. In
order to ensure that all possible combinations of di€erent
stereoisomers would be formed, one and/or two protect-
ing groups, which could also serve as lipophilic groups
and could be removed at the end of the synthesis to gen-
erate another more hydrophilic library of compounds,
were introduced ®rst on allyl d-xyloside.
extensive chromatographic work allowed us to isolate
easily a set of three monobenzylated derivatives 3 and 6
respectively and another set of dibenzylated compounds
2 and 7. Because the reactivity of the three hydroxyl
groups of xylose is not exactly identical, the amount of
each isomer in each mixture was not the same. This
ratio was corrected in order to work with equal
amounts of each product in each mixture. Careful
separation of the products yielded pure isomers which
were remixed in equal amounts or used to adjust the
composition of the original mixture checked by 1H
NMR. The six di€erent libraries prepared by this route
contained respectively the a-trihydroxy (4), a-dihydroxy
(3), a-monohydroxy derivatives (2) and the correspond-
ing b ones (5, 6 and 7).19 Fourteen di€erent products
were prepared this way and used in the following syn-
thetic steps in six di€erent reaction vessels (Scheme 1).
The carboxylic acid residue precursors were introduced
by alkylation of the free hydroxyl groups with tert-
butylbromoacetate to provide the six libraries 8±13.
Judicious choice of the components of the libraries,
containing only compounds of similar polarity, allowed
an easy puri®cation of each library by standard column
chromatography. The time was set to prepare the
deconvolution steps. The orthogonal design20 and
recursive deconvolution strategies21 were chosen as a
rapid, non-chemical way to identify active compounds.
Thus after storage of a portion (30%) of the libraries
(8±13), ®ve new libraries were constructed as follows.
All compounds of a con®guration were associated in
equal amounts in the ``a library'' 14. The same was done
for the b compounds giving the ``b library'' 15. Three
new libraries, orthogonal to the two previous ones, were
then constructed by mixing compounds containing three
(16), two (17), and one (18) ester residues respectively,
whatever the anomeric con®guration. Thus the fourteen
di€erent compounds were split in ®ve di€erent libraries,
each compound being present in two di€erent libraries
and only two (Fig. 3).
d-Xylose (1) was converted to known allyl d-xylosides
under standard conditions.18 The two anomers were
easily separated after acetylation. Subsequent deacety-
lation a€orded the free compounds 4 and 5, which were
separately benzylated using sodium hydride in DMF
and di€erent stoichiometries of benzyl bromide. This
allowed an almost random benzylation from which
Scheme 1. Reagents: i: (1) Allyl alcohol, Dowex H+, re¯ux, (2) Ac2O, pyridine then separation, (3) MeONa, MeOH; ii: NaH, BnBr, DMF, rt;
iii: NaH, BrCH2COOtBu, DMF.

514
N. Moitessier et al. / Bioorg. Med. Chem. 9 (2001) 511±523
After storage of a portion (30%) of the libraries (14±
18), parallel synthesis was carried out for the introduc-
tion of the amine residue. Ozonolysis of the allylic dou-
ble bond followed by NaBH4 reduction a€orded the
corresponding alcohols (19±23). Treatment of the latter
with triphenylphosphine and N-bromosuccinimide gave
the corresponding bromides (24±28) in good to excellent
yield (Scheme 2). Monitoring of these reactions was
easily performed by 13C NMR. A portion of the ®ve
mixtures of bromides was set aside and the new library
29 of fourteen di€erent bromo derivatives was prepared
by mixing the ®ve preceding mixtures in equal amounts.
This ®nal mixture 29 of fourteen bromides was split
into nine di€erent reaction vessels and treated in paral-
lel with nine di€erent primary amines in an SN2 reaction
followed by BOC protection of the resulting secondary
amine to facilitate puri®cation. This provided a library
of 126 compounds split into nine mixtures 30a±i. The
di€erent amino groups, which brought chemical diver-
sity to the library, were selected from literature pre-
cedents (Fig. 4). Finally libraries 30a±i were treated with
tri¯uoroacetic acid to remove all protecting groups
giving nine di€erent mixtures 31a±i.
Preliminary biological testings and deconvolution
The biological evaluation of these nine mixtures was
carried out by estimating the adhesion of S180 sarcoma
cells, which express only avb3 integrin, on a substrate of
®bronectin or vitronectin in the presence of the com-
pounds and compared to the e€ect of the RGDS peptide
as reference on the same cells. Mixtures 31a,b,d and i
showed signi®cant adhesion inhibitory activities (Fig. 5).
In particular compounds having an N-propyl substituent
contained in library 31a were active and were thus
further investigated to identify the most active com-
pound among the fourteen di€erent possible products.
The previously stored ®ve mixtures 24±28 of bromi-
nated derivatives were treated individually with propyl-
amine to provide the corresponding ®ve orthogonal
sub-libraries of compounds 32±36. (Scheme 3) These
®ve mixtures were tested as above and, among others,
two sub-libraries 32 containing a compounds and 36
containing dibenzylated derivatives were found active
(Fig. 6). Obviously the active molecule should be of a
con®guration and should have two benzyl groups. Only
three compounds have these characteristics (Fig. 7).
Normally the synthesis of the three pure compounds
would give the expected most active structure. In order
Figure 3. Construction of the orthogonal libraries 14, 15/16±18 by
mixing sub-libraries 8±13.
Scheme 3. Reagents: i: (1) PrNH2, DMF, 70 ꢀC, 2 h; (2) BOC2O,
NEt3, CH2Cl2; (3) TFA:H2O, 3:1.
Scheme 2. Reagents: i: (1) O3, MeOH/THF, À70 ꢀC; (2) NaBH4;
ii: PPh3, NBS, DMF, 50 ꢀC; iii: R5NH2, DMF, 70 ꢀC; iv: BOC2O,
NEt3, CH2Cl2; v: TFA:H2O, 3:1.
Figure 4. Structures of the di€erent amines used for libraries con-
struction.

N. Moitessier et al. / Bioorg. Med. Chem. 9 (2001) 511±523
515
Figure 5. Inhibition of cell adhesion on vitronectin substratum at 1 mg/mL of libraries 31a±i.
Figure 6. Inhibition of cell adhesion on vitronectin substratum at
2 mg/mL of orthogonal libraries 32±36.
Scheme 4. Reagents and conditions: i: (1) O3, MeOH/THF-70 ꢀC; (2)
NaBH4; ii: PPh3, NBS, DMF 50 ꢀC; iii: C3H7NH2, DMF, 70 ꢀC; iv:
BOC2O, NEt3, CH2Cl2; v: TFA:H2O, 3:1.
In conclusion we have presented one of the ®rst example
of carbohydrate-based libraries of peptidomimetics
using the stereodiversity concept. This stereodiversity,
provided by the use of a single judiciously substituted
carbohydrate sca€old, is an important feature of our
library and it represents an excellent way to mimic dif-
ferent conformations of the same peptide chain. Only
one carbohydrate sca€old was used in this study, the
stereodiversity being simply introduced by playing with
the di€erent hydroxyl groups. It is clear that the use of
di€erent carbohydrate sca€olds would increase the pos-
sibilities of our approach. A new dimension should be
introduced in combinatorial chemistry by emphasizing
stereochemical and regiochemical diversity rather than
chemical diversity. Some compounds with modest activ-
ities as antagonists of avb3 integrin have been identi®ed.
Further biological testings of the libraries are underway.
Having established the validity of this approach we are
currently applying this concept to the construction of
other biologically relevant libraries of peptidomimetics
and in the design of synthetic receptors using parallel
synthesis.
Figure 7. Final active mixture (39 was the active compound).
to shorten the deconvolution process we decided to turn
back to the theoretical model of the RGD sequence and
we checked which of the three compounds 37, 38 and 39
would have a chance to adopt a conformation ®tting
with the model. Molecular dynamics calculations clearly
showed that the compound 39 having the carboxylic
acid grafted at position 4 was the best candidate. Its
synthesis was then carried out from the corresponding
allyl 2,3-di-O-benzyl-a-d-xyloside (40) in 6 steps in an
overall yield of 76% (Scheme 4). The biological activity
of this compound in this test was identical to that of
RGDS, both compounds being tested at a 2 mg/mL
concentration (ca. 4 mM).
Experimental
General indications
All reactions were performed under nitrogen atmo-
sphere. Melting points are uncorrected and recorded
with a Buchi capillary tube melting-point apparatus.
Optical rotations were measured on a Perkin Elmer 141

516
N. Moitessier et al. / Bioorg. Med. Chem. 9 (2001) 511±523
polarimeter in a 1 dm cell at 20 ꢀC. FTIR spectra were
recorded on Perkin Elmer Spectrum 1000 on NaCl
graphed (H:E, 4:1 then 1:1 then 2:3) to provide the
monobenzylated derivatives (3.93 g, 13.6mmol, 49%),
and the three dibenzylated compounds (1.1g, 3 mmol,
11%) as well as some of the tribenzylated derivative
(0.25 g, 0.6 mmol, 2%).
1
windows or KBr pellets. H (250 MHz) and 13C NMR
(62.5 MHz) spectra were recorded on a Bruker AC 250
spectrometer. Unless otherwise stated, all spectra were
recorded in CDCl3. Chemical shifts are reported in ppm
using the residual of chloroform as internal standard
(7.27 ppm and 77.0 ppm respectively). Attribution of 13C
signals are based on the J-modulated spin-echo sequence.
Mass spectra were recorded on a Trio 1000 Thermo
Quest spectrometer in the electron impact mode or a
Platform Micromass in the electrospray mode. Elemental
analyses were obtained from the Service Central de
Microanalyse du CNRS, Vernaison (France). Analytical
thin-layer chromatography was performed on Merck
60F254 pre-coated silica gel plates. Preparative chro-
matography was performed on silica gel 60 (230-40
mesh ASTM) using ethyl acetate (E) and hexane (H)
mixtures.
Each pure component of mixture 2 was prepared and
characterized.
Allyl 2,3-di-O-benzyl-ꢀ-D-xylopyranoside. Rf=0.19
(H:E, 4:1); [a]D +44.7 (c 1.0, CHCl3); IR: 3436 cmÀ1
;
9
1H NMR: d 2.52 (s, 1H, OH), 3.52 (dd, 1H, J4,5 4, J5,5
Hz, H-5), 3.63 (m, 2H, H-2, H-3), 3.77 (ddd, 1H, J3,4
0
0
0
8.5, J4,5 9 Hz, H-4), 4.01 (ddd, 1H, J6,6 13, J6,7 6, J6,8
0
1
Hz, H-6), 4.04 (dd, 1H, H-50), 4.23 (ddd, 1H, J66 ,7 5 Hz,
J6 ,8 1 Hz, H-60), 4.68 (d, 1H, Jgem 12 Hz, CH2Ph), 4.71
(s, 2H, CH2Ph), 4.78 (d, 1H, J1,2 4 Hz, H-1), 4.96 (d,
0
0
1H, CH2Ph), 5.24 (dd, 1H, J7,8 10.5, J8,8 1.5 Hz, H-8),
0
5.33 (dd, 1H, J7,8 17 Hz, H-80), 5.95 (m, 1H, H-7), 7.35
(m, 10H, Ph); 13C NMR: d 61.9 (1C, C-5), 68.0 (1C, C-
6), 69.3 (1C, C-4), 72.8, 74.7 (2C, CH2Ph), 79.0 (1C, C-
2), 80.9 (1C, C-3), 95.7 (1C, C-1), 117.7 (1C, C-8), 127.6,
127.7, 128.1 (10C, CH Ph), 133.6 (1C, C-7), 137.8, 138.5
(2C, C Ph). Anal. calcd for C22H26O5: C, 71.3; H, 7.1.
Found: C, 71.71; H, 7.21.
Computational details. All calculations were performed
with the Insight 97.0/Discover package using the CVFF
force ®eld on Silicon Graphics1 Indigo2 workstations.
Molecular dynamics simulations were carried out using
a time step of 1 fs. Optimizations were performed by
conjugate gradient energy minimization with a con-
vergence criterion of 0.001 kcal/mol. Atom partial
charges were calculated using MNDO semiempirical
method by calculation of the Mulliken electronic popu-
lation. For in vacuo simulations, solvent conditions were
represented implicitly using a distance-dependent dielectric
constant E=r. For dynamics simulations with explicit
water molecules, periodic boundary conditions were
applied, using water model provided by MSI and a ®xed
dielectric constant of 1.0. Conformational samplings and
molecular dynamics in explicit solvent were performed
according to the previously reported procedures.9b
Allyl 2,4-di-O-benzyl-ꢀ-D-xylopyranoside. Rf=0.28
1
(H:E, 4:1); [a]D +72.2 (c 0.7, CHCl3); H NMR: d 2.58
0
(d, 1H, JH,OH 2 Hz, OH), 3.34 (dd, 1H, J4,5 4, J5,5 9 Hz,
0
H-5), 3.53 (m, 3H, H-2, H-3, H-4), 3.92 (ddd, 1H, J6,6
0
13, J6,7 6, J6,8 1 Hz, H-6), 4.04 (dd, 1H, J4,5 9 Hz, H-50),
4.13 (ddd, 1H, J6 ,7 5, J6 ,8 1 Hz, H-60), 4.64 (2d, 2H,
Jgem 12 Hz, CH2Ph), 4.67 (d, 1H, J1,2 4 Hz, H-1), 4.76
(d, 1H, CH2Ph), 4.78 (d, 1H, CH2Ph), 5.21 (dd, 1H, J7,8
0
0
10.5, J8,8 1.5 Hz, H-8), 5.32 (dd, 1H, J7,8 17 Hz, H-80),
5.90 (m, 1H, H-7), 7.33 (m, 10H, Ph); 13C NMR: d 59.3
(1C, C-5), 67.7 (1C, C-6), 72.2 (1C, C-3), 72.4, 72.7 (2C,
CH2Ph), 77.2 (1C, C-4), 79.0 (1C, C-2), 95.0 (1C, C-1),
117.7 (1C, C-8), 127.31, 127.35, 127.43, 127.57, 127.93,
127.97, 128.00 (10C, CH Ph), 133.4 (1C, C-7), 137.7,
138.0 (2C, C Ph).
0
0
Cell cultures and reagents. The sarcoma 180 cells used in
this study were a sub-clone derived by Dr. K. Yamada
(NIH, Bethesda, USA) from the original parental cell
line, which was obtained from ATCC (CCL8; CCRF
S180 II). These cells were selected for their inability to
assemble a ®bronectin (FN) matrix at the cell surface.
They were cultured in Dubelcco's modi®ed Eagle's
medium (DMEM) containing 10% (v/v) fetal calf serum
(FCS), penicillin (100 I.U./mL)±streptomycin (100 mg/
mL) and 2 mM l-glutamine (Seromed) in a 37 ꢀC incu-
bator under an atmosphere of 6% CO2/94% air. Fibro-
nectin, vitronectin and RGDS peptides were purchased
from Sigma.
Allyl 3,4-di-O-benzyl-ꢀ-D-xylopyranoside. Rf=0.26
1
(H:E, 4:1); [a]D +90.6 (c 0.5, CHCl3); H NMR: d 2.29
(d, 1H, JH,OH 8 Hz, OH), 3.55 (bm, 3H, H-3, H-4, H-5),
0
3.69 (bm, 1H, H-2), 4.02 (dd, 1H, J6,6 13, J6,7 6, H-6),
0
0
0
0
4.04 (dd, 1H, J4,5 9 Hz, H-5 ), 4.22 (dddd, 1H, J6 ,7 5, J6 ,8
0
0
0
1, J6 ,8 1 Hz, H-6 ), 4.63 (d, 1H, Jgem 12 Hz, CH2Ph), 4.73
(d, 1H, CH2Ph), 4.85 (d, 1H, J1,2 4 Hz, H-1), 4.85 (d, 1H,
Jgem 12 Hz, CH2Ph), 4.91 (d, 1H, CH2Ph), 5.21 (dd, 1H,
0
0
J7,8 10.5, J8,8 1.5 Hz, H-8), 5.31 (dd, 1H, J7,8 17 Hz, H-
80), 5.92 (m, 1H, H-7), 7.33 (m, 10H, Ph); 13C NMR: d
60.1 (1C, C-5), 68.1 (1C, C-6), 72.0 (1C, C-2), 72.9, 74.8
(2C, CH2Ph), 77.3 (1C, C-4), 81.6 (1C, C-3), 97.3 (1C,
C-1), 117.5 (1C, C-8), 127.3, 127.5, 127.53, 127.58,
128.08, 128.16 (10C, CH Ph), 133.5 (1C, C-7), 137.9,
138.5 (2C, C Ph).
Libraries construction: chemistry
Non selective benzylation. NaH (1.11 g, 27.7 mmol)
was suspended in DMF (80 mL) and allyl d-xyloside
(5.25 g, 27.7 mmol) in DMF (100 mL) was added drop-
wise at 0 ꢀC. After stirring for 30 min, benzyl bromide
(3.3 mL, 27.7 mmol) was added in one portion at room
temperature. The resulting mixture was stirred for 16 h
then quenched with methanol (10 mL) at 0 ꢀC. The
solution was concentrated in vacuo then taken up in
dichloromethane, washed with water, dried over MgSO4
and concentrated in vacuo. The residue was chromato-
Mixture 3
Allyl 2-O-benzyl-ꢀ-D-xylopyranoside. Rf=0.60 (H:E,
1
1:4); [a]D +107.2 (c 0.9, CHCl3); H NMR: d 2.59 (bs,
1H, OH), 2.72 (bs, 1H, OH), 3.32 (dd, 1H, J1,2 3.5, J2,3

N. Moitessier et al. / Bioorg. Med. Chem. 9 (2001) 511±523
517
10 Hz, H-2), 3.54 (m, 3H, H-4, H-5, H-50), 3.87 (dd, 1H,
JH,OH 2.5 Hz, OH), 3.26 (dd, 1H, J4,5 9.5, J5,5 12 Hz, H-
5), 3.38 (dd, 1H, J2,3=J3,4 8 Hz, H-3), 3.57 (ddd, 1H,
0
0
J3,4 5 Hz, H-3), 3.91 (dd, 1H, J6,6 13, J6,7 6 Hz, H-6),
0
4.15 (dd, 1H, J6 ,7 5 Hz, H-60), 4.60 (d, 1H, Jgem 12 Hz,
CH2Ph), 4.68 (d, 1H, CH2Ph), 4.76 (d, 1H, H-1), 5.21
J1,2 8 Hz, H-2), 3.73 (ddd, 1H, J4,5 5 Hz, H-4), 4.02 (dd,
0
0
1H, H-50), 4.12 (dd, 1H, J6,6 13, J6,7 6 Hz, H-6), 4.31 (d,
(dd, 1H, J7,8 10.5, J8,8 1.5 Hz, H-8), 5.32 (dd, 1H, J7,8
1H, H-1), 4.36 (dd, 1H, J6 ,7 5 Hz, H-60), 4.74 (d, 1H,
Jgem 12 Hz, CH2Ph), 5.00 (d, 1H, CH2Ph), 5.23 (dd, 1H,
0
0
0
17 Hz, H-80), 5.89 (m, 1H, H-7), 7.33 (m, 5H, Ph); 13C
NMR: d 61.3 (1C, C-5), 67.8 (1C, C-6), 69.8 (1C, C-4),
72.6 (1C, CH2Ph), 72.9 (1C, C-2), 79.1 (1C, C-3), 95.4
(1C, C-1), 116.8 (1C, C-8), 127.6, 127.8, 128.1 (5C, CH
Ph), 133.6 (1C, C-7), 137.9 (1C, C Ph). EIMS: m/z:
280.7 (M)+ꢁ , 91 (Bn). Anal. calcd for C15H20O5: C,
64.3; H, 7.2. Found: C, 64.06%; H, 7.13%.
0
0
J7,8 10.5, J8,8 1.5 Hz, H-8), 5.33 (dd, 1H, J7,8 16.5 Hz,
H-80), 5.94 (m, 1H, H-7), 7.34 (m, 5H, Ph); 13C NMR: d
65.0 (1C, C-5), 69.2 (1C, C-4), 70.1 (1C, C-6), 73.4 (1C,
C-2), 74.4 (1C, CH2Ph), 83.10 (1C, C-3), 102.30 (1C, C-
1), 118.03 (1C, C-8), 127.9, 128.1, 128.6 (5C, CH Ph),
133.9 (1C, C-7), 138.6 (1C, C Ph).
Allyl 3-O-benzyl-ꢀ-D-xylopyranoside. Rf=0.63 (H:E,
1:4); mp 92 ꢀC (CH2Cl2/H); [a]D +68.9 (c 1.5, CHCl3);
1H NMR: d 2.29 (d, 1H, JH,OH 7.5 Hz, OH), 2.56 (s, 1H,
OH), 3.66 (m, 5H, H-2, H-3, H-4, H-5, H-50), 4.04 (dd,
Allyl 4-O-benzyl-ꢁ-D-xylopyranoside. Rf=0.38 (H:E,
2:3); [a]D À77.1 (c 1.2, CHCl3); 1H NMR: d 3.14 (d, 1H,
JH,OH 6.5 Hz, OH), 3.21 (d, 1H, JH,OH 7 Hz, OH), 3.50
(m, 1H, H-4), 3.55 (m, 2H, H-2, H-5), 3.82 (ddd, 1H,
0
0
0
0
1H, J6,6 13, J6,7 6.5 Hz, H-6), 4.30 (dd, 1H, J6 ,7 5.5 Hz,
H-60), 4.70 (d, 1H, Jgem 11.5 Hz, CH2Ph), 4.80 (d, 1H,
J1,2 3 Hz, H-1), 4.94 (d, 1H, CH2Ph), 5.23 (dd, 1H, J7,8
J2,3=J3,4 8 Hz, H-3), 4.02 (dd, 1H, J4,5 5, J5,5 12 Hz,
0
H-50), 4.08 (dd, 1H, J6,6 13, J6,7 6 Hz, H-6), 4.30 (dd,
1H, J6 ,7 5.5 Hz, H-60), 4.60 (d, 1H, J1,2 4.5 Hz, H-1),
0
0
0
0
10.5 Hz, J8,8 1.5 Hz, H-8), 5.32 (dd, 1H, J7,8 15.5 Hz,
H-80), 5.92 (m, 1H, H-7), 7.35 (m, 5H, Ph); 13C NMR: d
62.7 (1C, C-5), 68.5 (1C, C-6), 68.7 (1C, C-4), 71.8 (1C,
C-2), 74.2 (1C, CH2Ph), 81.6 (1C, C-3), 97.5 (1C, C-1),
117.8 (1C, C-8), 127.7, 128.4 (5C, CH Ph), 133.5 (1C, C-
7), 138.2 (1C, C Ph). Anal. calcd for C15H20O5: C, 64.3;
H, 7.2. Found: C, 64.40; H, 7.04.
4.68 (s, 2H, CH2Ph), 5.23 (dd, 1H, J7,8 11, J8,8 1.5 Hz,
0
H-8), 5.30 (dd, 1H, J7,8 17.5 Hz, H-80), 5.92 (m, 1H, H-
7), 7.35 (m, 5H, Ph); 13C NMR: d 62.3 (1C, C-5), 69.5
(1C, C-6), 71.5 (1C, CH2Ph), 72.3 (1C, C-2), 73.7 (1C,
C-3), 76.7 (1C, C-4), 101.4 (1C, C-1), 117.6 (1C, C-8),
127.5, 127.6, 128.0 (5C, CH Ph), 133.5 (1C, C-7), 137.7
(1C, C Ph).
Allyl 4-O-benzyl-ꢀ-D-xylopyranoside. Rf=0.60 (H:E, 1:4);
1H NMR: d 2.23 (d, 1H, JH,OH 7 Hz, OH), 2.78 (bs, 1H,
OH), 3.50 (m, 3H, H-2, H-5, H-50), 3.68 (m, 1H, H-4),
Mixture 7
Allyl 2,3-di-O-benzyl-ꢁ-D-xylopyranoside. Rf=0.29
(H:E, 4:1); mp 67 ꢀC (CH2Cl2/H); [a]D À32.7 (c 0.6,
0
3.82 (dd, 1H, J2,3=J3,4 8 Hz, H-3), 3.99 (dd, 1H, J6,6
0
13, J6,7 6 Hz, H-6), 4.21 (dd, 1H, J6 ,7 5 Hz, H-60), 4.64
(d, 1H, Jgem 12 Hz, CH2Ph), 4.72 (d, 1H, CH2Ph), 4.84 (d,
CHCl3); H NMR: d 2.31 (d, 1H, JH,OH 4.5 Hz, OH),
1
0
3.26 (dd, 1H, J4,5 8.5, J5,5 11.5 Hz, H-5), 3.45 (dd, 1H,
0
1H, J1,2 4.5 Hz, H-1), 5.21 (dd, 1H, J7,8 10.5, J8,8 1.5 Hz,
0
J2,3 5, J3,4 5 Hz, H-3), 3.67 (m, 1H, H-4), 4.01 (dd, 1H,
0
H-8), 5.28 (dd, 1H, J7,8 17 Hz, H-80), 5.91 (m, 1H, H-7),
J4,5 5 Hz, H-5 ), 4.11 (ddd, 1H, J6,6 13, J6,7 6, J6,8 0.5 Hz,
0
0
7.33 (m, 5H, Ph); 13C NMR: d 59.6 (1C, C-5), 67.7 (1C, C-
6), 71.7 (1C, C-2), 72.6 (1C, CH2Ph), 73.1 (1C, C-3), 77.2
(1C, C-4), 97.0 (1C, C-1), 117.0 (1C, C-8), 127.2, 127.3,
127.8 (5C, CH Ph), 133.5 (1C, C-7), 138.0 (1C, C Ph).
H-6), 4.37 (ddd, 1H, J6 ,7 6, J6 ,8 0.5 Hz, H-60), 4.50 (dd,
J1,2 5 Hz, H-2), 4.61 (d, 1H, H-1), 4.63 (d, 1H, Jgem 12 Hz,
CH2Ph), 4.68 (d, 1H, CH2Ph), 4.90 (d, 1H, Jgem 12 Hz,
0
0
0
CH2Ph), 4.91 (d, 1H, CH2Ph), 5.21 (ddd, 1H, J7,8 10, J8,8
0
0.5 Hz, H-8), 5.30 (dd, 1H, J7,8 18 Hz, H-80), 5.92 (m,
1H, H-7), 7.33 (m, 10H, Ph); 13C NMR: d 64.5 (1C, C-
5), 69.1 (1C, C-4), 69.7 (1C, C-6), 74.1, 74.4 (2C,
CH2Ph), 80.4 (1C, C-2), 82.5 (1C, C-3), 102.3 (1C, C-1),
117.1 (1C, C-8), 127.54, 127.57, 127.65, 127.71, 127.90,
128.18, 128.27 (10C, CH Ph), 133.7 (1C, C-7), 138.0,
138.3 (2C, C Ph). Anal. calcd for C23H28O4: C, 71.3; H,
7.1. Found: C, 71.16; H, 7.23.
Mixture 6
Allyl 2-O-benzyl-ꢁ-D-xylopyranoside. Rf=0.33 (H:E, 2:3);
mp 64ꢀC (CH2Cl2/H); [a]D À11.2 (c 2.8, CHCl3); H
1
0
NMR: d 3.09 (bs, 2H, OH), 3.37 (dd, 1H, J4,5 9.5, J5,5
11.5 Hz, H-5), 3.40 (dd, 1H, J1,2 7.5, J2,3 5.5 Hz, H-2),
3.64 (m, 2H, H-3, H-4), 4.01 (dd, 1H, J4,5 3.5 Hz, H-50),
0
0
4.08 (dd, 1H, J6,6 13, J6,7 6.5 Hz, H-6), 4.33 (dddd, 1H,
0
J6 ,7 5, J6 ,8 1, J6 ,8 1 Hz, H-60), 4.61 (d, 1H, H-1), 4.65
(d, 1H, Jgem 11.5 Hz, CH2Ph), 4.84 (d, 1H, CH2Ph),
Allyl 2,4-di-O-benzyl-ꢁ-D-xylopyranoside. Rf=0.40
1
0
0
0
(H:E, 4:1); [a]D +1.8 (c 1.6, CHCl3); H NMR: d 2.53
0
0
(d, 1H, JH,OH 2.5 Hz, OH), 3.22 (dd, 1H, J4,5 10, J5,5 12
5.22 (ddd, 1H, J7,8 11, J8,8 1.5 Hz, H-8), 5.31 (ddd, 1H,
0
J7,8 17.5 Hz, H-80), 5.92 (m, 1H, H-7), 7.33 (m, 5H, Ph);
Hz, H-5), 3.27 (dd, 1H, J1,2 7.5, J2,3 9 Hz, H-2), 3.51 (ddd,
0
1H, J3,4 9, J4,5 5 Hz, H-4), 3.68 (m, 1H, H-3), 3.93 (dd,
13C NMR: d 64.7 (1C, C-5), 69.2 (1C, C-4), 69.4 (1C, C-
6), 73.8 (1C, CH2Ph), 75.0 (1C, C-3), 80.3 (1C, C-2),
102.2 (1C, C-1), 116.9 (1C, C-8), 127.3, 127.6, 127.8
(5C, CH Ph), 133.5 (1C, C-7), 138.0 (1C, C Ph). Anal.
calcd for C15H20O5: C, 64.3; H, 7.2. Found: C, 64.16; H,
7.12.
1H, H-50), 4.11 (ddd, 1H, J6,6 12.5, J6,7 6, J6,8 0.5 Hz, H-
0
6), 4.37 (ddd, 1H, J6 ,7 5, J6 ,8 0.5 Hz, H-60), 4.40 (d, 1H, H-
1), 4.62 (d, 1H, Jgem 12 Hz, CH2Ph), 4.68 (d, 1H, CH2Ph),
4.76 (d, 1H, Jgem 12 Hz, CH2Ph), 4.94 (d, 1H, CH2Ph),
0
0
0
5.22 (ddd, 1H, J7,8 10.5, J8,8 0.5 Hz, H-8), 5.33 (dd, 1H,
0
J7,8 17.5 Hz, H-80), 5.94 (m, 1H, H-7), 7.34 (m, 10H,
Allyl 3-O-benzyl-ꢁ-D-xylopyranoside. Rf=0.63 (H:E, 1:1);
mp 96ꢀC (CH2Cl2/H); [a]D À56.9 (c 0.6, CHCl3); H
Ph); 13C NMR: d 63.5 (1C, C-5), 69.7 (1C, C-6), 72.7,
74.1 (2C, CH2Ph), 75.2 (1C, C-3), 76.9 (1C, C-4), 80.8
(1C, C-2), 102.5 (1C, C-1), 117.0 (1C, C-8), 127.44,
1
NMR: d 2.20 (d, 1H, JH,OH 3 Hz, OH), 2.43 (d, 1H,

518
N. Moitessier et al. / Bioorg. Med. Chem. 9 (2001) 511±523
127.51, 127.75, 128.12, 128.15 (10C, CH Ph), 133.7 (1C,
C-7), 138.0, 138.2 (2C, C Ph).
Libraries characterization
All mixtures were characterized by 13C NMR. Obviously,
due to signal overlap, the number of peaks assigned to a
group of identical carbon atoms can di€er from the
expected one. (See Scheme 4 for carbon numbering.)
Allyl 3,4-di-O-benzyl-ꢁ-D-xylopyranoside. Rf=0.32
1
(H:E, 4:1); [a]D À11.3 (c 0.9, CHCl3); H NMR: d 2.63
0
(d, 1H, JH,OH 2.5 Hz, OH), 3.32 (dd, 1H, J4,5 8, J5,5 12
Hz, H-5), 3.59 (m, 3H, H-2, H-3, H-4), 4.02 (dd, 1H,
J4,5 4.5 Hz, H-50), 4.12 (dd, 1H, J6,6 13, J6,7 6.5 Hz, H-
0
0
Mixture 8 (3 compounds). 92%; Rf=0.55±0.70 (H:E,
4:1); 13C NMR: d 27.47, 27.53, 27.60 (3Â3C, OtBu), 59.53,
59.63, 59.64 (3Â1C, C-5), 67.51, 67.70, 67.88 (3Â1C, C-6),
69.04, 69.25, 70.79, 72.57, 72.73, 72.78, 72.94, 75.12
(3Â3C, CH2Ph, CH2COOtBu), 77.15, 77.95, 78.70, 78.99,
79.36, 80.11, 81.22, 81.31, 82.20 (3Â3C, C-2, C-3, C-4),
80.43, 80.85, 81.06 (3Â1C, OtBu), 95.09, 95.26, 96.38
(3Â1C, C-1), 116.98, 117.23, 117.37 (3Â1C, C-7), 127.09,
127.13, 127.19, 127.29, 127.47, 127.56, 127.85, 127.89
(3Â10C, CH Ph), 133.38, 133.42, 133.57 (3Â1C, C-8),
137.82, 137.86, 137.98, 138.35, 138.45 (3Â2C, C Ph),
168.86, 168.91, 169.21 (3Â1C, COOtBu).
6), 4.35 (dd, 1H, J6 ,7 5 Hz, H-60), 4.40 (d, 1H, J1,2 6 Hz,
H-1), 4.64 (d, 1H, Jgem 12 Hz, CH2Ph), 4.72 (d, 1H,
0
CH2Ph), 4.86 (s, 2H, CH2Ph), 5.23 (dd, 1H, J7,8 10.5,
J8,8 1 Hz, H-8), 5.33 (dd, 1H, J7,8 17 Hz, H-80), 5.96 (m,
1H, H-7), 7.34 (m, 10H, Ph); 13C NMR: d 64.6 (1C, C-
5), 67.9 (1C, C-6), 72.5 (1C, C-2), 72.9, 74.2 (2C, CH2Ph),
76.7 (1C, C-4), 81.6 (1C, C-3), 101.7 (1C, C-1), 117.4 (1C,
C-8), 127.36, 127.54, 127.61, 127.73, 127.88, 128.08, 128.17
(10C, CH Ph), 133.7 (1C, C-7), 137.7, 138.3 (2C, C Ph).
0
0
Typical procedures
The following procedures have been used for the pre-
paration of libraries 31.
Mixture 9 (3 compounds). 84%; Rf=0.55±0.70 (H:E,
4:1); 13C NMR: d 27.54, 27.59, 27.66 (3Â6C, OtBu), 59.39,
59.55, 59.63 (3Â1C, C-5), 67.56, 67.74, 67.78 (3Â1C, C-6),
68.96, 69.10, 69.18, 69.22, 70.69, 70.80, 72.62, 73.60, 75.16
(3Â3C, CH2Ph, CH2COOtBu), 77.74, 77.96, 79.11, 79.27,
79.42, 80.22, 81.58, 82.58, 82.76 (3Â3C, C-2, C-3, C-4),
80.58, 80.68, 80.84, 80.97, 81.06, 81.13 (3Â2C, OtBu),
95.06, 95.24, 96.34 (3Â1C, C-1), 117.04, 117.12, 117.40
(3Â1C, C-7), 127.27, 127.34, 127.37, 127.54, 127.78,
127.98 (3Â5C, CHPh), 133.37, 133.54, 133.57 (3Â1C, C-
8), 137.84, 137.92, 138.25 (3Â1C, C Ph), 168.52, 168.88,
168.99, 169.03, 169.26, 169.50 (3Â2C, COOtBu).
Alkylation. NaH (849 mg, 21.2 mmol) was suspended in
DMF (50 mL) and allyl b-d-xyloside (1.00 g, 5.26mmol)
in DMF (50 mL) was added dropwise at 0 ꢀC. To this sus-
pension was subsequently added tert-butylbromoacetate
(5 mL). The resulting mixture was stirred for 16 h then
quenched with methanol (5 mL) at 0 ꢀC. The solution was
concentrated in vacuo then taken up in dichloromethane,
washed with water, dried over MgSO4 and concentrated
in vacuo. The residue was chromatographed (H:E, 9:1
then 4:1) to provide 11 (2.15 mg, 4.04mmol, 77%).
Compound 10. 77%; Rf=0.55 (H:E, 4:1); 13C NMR: d
28.01, 28.34 (9C, OtBu), 59.74 (1C, C-5), 68.14 (1C, C-6),
69.26, 69.39, 71.03 (3C, CH2COOtBu), 78.99 (1C, C-4),
79.99 (1C, C-2), 81.17, 81.42, 81.49 (3C, OtBu), 83.21
(1C, C-3), 96.36 (1C, C-1), 117.64 (1C, C-7), 133.71 (1C,
C-8), 169.31, 169.64, 169.71 (3C, COOtBu).
Ozonolysis. O3 was bubbled through a vigorously stirred
solution of the ole®n (1 mmol) in a mixture of MeOH:
THF (20 mL, 2:1) at À78 ꢀC until starting material dis-
appearance (TLC monitoring) then sodium borohydride
(168 mg, 1.06 mmol) was added. Puri®cation by chroma-
tography (H:E, 4:1 then 3:2) a€orded the alcohol (90±
96%).
Compound 11. 77%; Rf=0.59 (H:E, 4:1); 13C NMR: d
27.93, 27.96 (9C, OtBu), 63.8 (1C, C-5), 69.3 (1C, C-6),
69.8, 70.8 (3C, CH2COOtBu), 77.5 (1C, C-4), 80.8, 80.9,
81.1 (3C, OtBu), 82.8 (1C, C-2), 84.9 (1C, C-3), 102.4
(1C, C-1), 117.0 (1C, C-7), 133.8 (1C, C-8), 169.0, 169.3,
169.7 (3C, COOtBu)
Bromination. To a solution of the alcohol (1.0 mmol)
and N-bromosuccinimide (354 mg, 2 mmol) in DMF
was added triphenylphosphine (524 mg, 2 mmol). The
mixture was warmed up to 70 ꢀC and stirred for a fur-
ther 30 min. The reagent in excess was quenched with
methanol (2 mL) and the mixture was concentrated in
vacuo. The residue was chromatographed (H:E, 4:1) to
yield the bromo derivative (70±76%).
Mixture 12 (3 compounds). 80%; Rf=0.55±0.70 (H:E,
4:1); 13C NMR: d 27.69 (3Â6C, OtBu), 63.32, 63.43, 63.54
(3Â1C, C-5), 68.00, 68.73, 68.96 (3Â1C, C-6), 69.03,
69.60, 69.81, 70.64, 70.70, 73.01, 73.63, 74.20, 75.10
(3Â3C, CH2Ph, CH2COOtBu), 76.40, 77.86, 78.11, 81.28,
82.55, 82.87, 83.20, 84.42, 84.97 (3Â3C, C-2, C-3, C-4),
80.50, 80.73, 80.99, 81.06 (3Â2C, OtBu), 102.16, 102.30,
102.60 (3Â1C, C-1), 116.78, 116.83 (3Â1C, C-7), 127.20,
127.31, 127.44, 127.53, 127.63, 127.89, 127.97 (3Â5C, CH
Ph), 133.41, 133.58 (3Â1C, C-8), 138.00, 138.08, 138.33
(3Â1C, C Ph), 168.54, 168.76, 168.86, 168.99, 169.24,
169.38 (3Â2C, COOtBu).
Amination/protection. The bromo-derivative (1 mmol)
was re¯uxed in DMF (20 mL) in the presence of amine
(5 mL) for 30 min. After concentration in vacuo, the
crude product was treated by BOC2O (0.25 mL) and
triethylamine (0.14 mL) in dichloromethane (10 mL) for
1 h to a€ord the pure BOC protected derivative after
column chromatography (85±92%).
Deprotection. The protected compound was stirred in a
TFA:water mixture (3:1) for 16 h. The concentration in
vacuo, followed by dilution in water and lyophilization
led to the tri¯uoroacetate salts as foams (80±90%).
Mixture 13 (3 compounds). 96%; Rf=0.55±0.70 (H:E,
4:1); 13C NMR: d 27.94 (3Â3C, OtBu), 63.68, 63.74,
63.90 (3Â1C, C-5), 68.31, 69.31, 69.99, 70.06, 70.28,

N. Moitessier et al. / Bioorg. Med. Chem. 9 (2001) 511±523
519
71.05, 73.25, 74.69, 74.71, 75.42, 75.47 (3Â4C, C-6,
CH2Ph, CH2COOtBu), 77.20, 77.29, 78.88, 81.42, 81.80,
83.18, 83.86, 84.95 (3Â3C, C-2, C-3, C-4), 81.06, 81.69,
81.75 (3Â1C, OtBu), 102.58, 102.88, 103.07 (3Â1C, C-1),
117.10, 117.16, 117.19 (3Â1C, C-7), 127.49, 127.66, 127.88,
127.93, 128.02, 128.14, 128.18, 127.27 (3Â10C, CH Ph),
133.94 (3Â1C, C-8), 138.06, 138.16, 138.30, 138.41, 138.65
(3Â2C, C Ph), 169.04, 169.08, 169.46 (3Â1C, COOtBu).
82.93, 83.38, 83.74, 84.91, 85.28 (6Â3C, C-2, C-3, C-4),
81.47, 81.54, 81.61, 81.69, 82.83 (6Â2C, OtBu), 97.23,
97.94, 98.01, 103.59, 103.76, 104.26 (6Â1C, C-1), 127.67,
127.77, 127.98, 128.09, 128.16, 128.23, 128.35, 128.44
(6Â5C, CH Ph), 138.12, 138.40, 138.60, 138.75 (6Â1C, C
Ph), 169.26, 169.40, 169.59, 169.65, 169.86, 170.10
(6Â2C, COOtBu).
Mixture 23 (6 compounds). 91%; Rf=0.4±0.5 (H:E,
4:1); 13C NMR: d 28.33 (6Â3C, OtBu), 60.25, 60.31,
60.43, 64.05, 64.09, 64.30 (6Â1C, C-5), 61.89, 62.05, 62.26
(6Â1C, C-7), 69.69, 69.79, 70.35, 70.63, 70.98, 71.40, 71.51,
72.60, 72.73, 73.60, 73.71, 73.89, 74.08, 75.29, 75.84, 75.94
(6Â4C, C-6, CH2Ph, CH2COOtBu), 77.53, 77.87, 78.55,
79.17, 79.38, 79.44, 79.95, 81.29, 81.67, 81.81, 81.86, 82.12,
82.82, 83.79, 83.85, 84.27, 85.35 (6Â3C, C-2, C-3, C-4),
82.04, 82.20 (6Â1C, OtBu), 98.00, 98.15, 98.55, 103.99,
104.55, 104.75 (6Â1C, C-1), 127.97, 128.04, 128.07,
128.31, 128.39, 128.43, 128.67, (6Â10C, CH Ph), 138.05,
138.26, 138.38, 138.53, 138.63, 138.65, 138.83 (6Â2C, C
Ph), 169.43, 169.61, 169.85, 169.94, 170.07, 170.26
(6Â1C, COOtBu).
Mixtures 19±23 were obtained by ozonolysis/reduction
of libraries 14±18.
Mixture 19 (7 compounds). 84%; Rf=0.4±0.5 (H:E,
4:1); 13C NMR: d 27.56 (1Â9C, 3Â6C, 3Â3C, OtBu),
59.58, 59.77, 59.83, 59.93, 60.16 (7Â1C, C-5), 61.37
(7Â1C, C-7), 68.29, 68.97, 69.14, 69.25, 69.30, 69.63,
69.75, 69.84, 70.06, 70.86, 70.90, 70.97, 71.04, 72.96,
73.09, 73.22, 73.35, 73.54, 75.45 (7Â4C, C-6, CH2Ph,
CH2COOtBu), 77.44, 77.91, 78.09, 78.24, 78.83, 78.90,
79.00, 79.21, 79.27, 79.51, 80.03, 80.36, 80.81, 80.89,
81.37, 81.39, 81.61, 82.36, 82.63, 82.76, 82.92 (7Â3C, C-
2, C-3, C-4), 81.04, 81.13, 81.48, 81.69 (1Â3C, 3Â2C,
3Â1C, OtBu), 97.35, 97.44, 97.58, 97.75, 97.92, 98.05
(7Â1C, C-1), 127.57, 127.83, 127.95, 128.23, 128.30
(3Â10C, 3Â5C, CH Ph), 137.64, 137.81, 137.94, 138.11,
138.28, 138.40, 138.44 (3Â2C, 3Â1C, C Ph), 169.15,
169.19, 169.34, 169.41, 169.47, 169.66, 169.71, 169.80,
169.96 (1Â3C, 3Â2C, 3Â1C, COOtBu).
Bromination
Mixture 24 (7 compounds). 88%; Rf=0.4±0.55 (H:E,
4:1); 13C NMR: d 27.82 (1Â9C, 3Â6C, 3Â3C, OtBu),
29.57 (7Â1C, C-7), 59.86, 59.98, 60.08 (7Â1C, C-5), 67.63,
67.84, 68.99, 69.10, 69.27, 69.33, 69.51, 69.63, 70.84, 70.96,
71.00, 72.95, 73.14, 73.19, 75.39 (7Â4C, C-6, CH2Ph,
CH2COOtBu), 77.15, 77.75, 77.97, 78.67, 78.72, 79.10,
79.36, 79.47, 79.87, 80.33, 80.40, 81.20, 81.48, 82.17,
82.48, 82.64, 82.80 (7Â3C, C-2, C-3, C-4), 80.85, 80.94,
81.14, 81.24, 81.31 (1Â3C, 3Â2C, 3Â1C, OtBu), 96.73,
96.89, 96.99, 97.51, 97.74, 97.90 (7Â1C, C-1), 127.38,
127.50, 127.62, 127.75, 127.91, 128.15 (3Â10C, 3Â5C, CH
Ph), 137.92, 138.08, 138.27, 138.40, 138.47 (3Â2C, 3Â1C,
C Ph), 169.07, 169.27, 169.39, 169.42, 169.52, 169.63,
169.78 (1Â3C, 3Â2C, 3Â1C, COOtBu).
Mixture 20 (7 compounds). 95%; Rf=0.4±0.5 (H:E,
4:1); 13C NMR: d 28.11 (1Â9C, 3Â6C, 3Â3C, OtBu),
61.83, 62.05 (7Â1C, C-7), 63.71, 63.82, 63.85, 63.91, 63.99,
64.07 (7Â1C, C-5), 69.38, 69.41, 69.44, 70.42, 70.61, 70.74,
70.99, 71.07, 71.15, 72.37, 72.52, 72.57, 73.32, 73.35, 74.87,
75.01, 75.04, 75.54, 75.57 (7Â4C, C-6, C-9, CH2Ph,
CH2COOtBu), 77.21, 77.29, 77.61, 78.14, 78.27, 78.77,
78.89, 78.94, 79.05, 81.43, 81.55, 81.88, 83.01, 83.08,
83.52, 83.56, 83.64, 83.79, 84.03, 85.04, 85.12, 85.25,
85.36 (7Â3C, C-2, C-3, C-4), 81.30, 81.35, 81.66, 81.75
(1Â3C, 3Â2C, 3Â1C, OtBu), 103.59, 103.76, 104.31,
104.53 (7Â1C, C-1), 127.64, 127.73, 127.80, 127.84, 127.87,
127.91, 127.96, 128.01, 128.15, 128.32, 128.35, 128.43
(3Â5C, 3Â10C, CH Ph), 138.03, 138.09, 138.14, 138.28,
138.37, 138.40, 138.43, 138.46, 138.57 (3Â1C, 3Â2C, C
Ph), 169.14, 169.33, 169.53, 169.69, 169.72, 169.83,
169.96 (1Â3C, 3Â2C, 3Â1C, COOtBu).
Mixture 25 (7 compounds). 88%; Rf=0.4±0.55 (H:E,
4:1); 13C NMR: d 27.45 (1Â9C, 3Â6C, 3Â3C, OtBu),
29.55, 29.61, 29.70, 29.74 (7Â1C, C-7), 63.33, 63.39,
63.53 (7Â1C, C-5), 68.84, 68.98, 69.02, 69.50, 69.70,
69.86, 70.46, 70.52, 70.56, 70.61, 72.80, 72.84, 72.94,
74.11, 74.26, 74.28, 75.00, 75.04, (7Â4C, C-6, CH2Ph,
CH2COOtBu), 76.63, 76.69, 77.00, 77.58, 77.80, 78.32,
80.78, 80.88, 81.13, 82.03, 82.42, 82.57, 82.82, 83.27,
84.10, 84.34, 84.39, 84.61 (7Â3C, C-2, C-3, C-4), 80.61,
80.67, 80.75, 80.96, 81.01, 81.10 (1Â3C, 3Â2C, 3Â1C,
OtBu), 103.05, 103.20, 103.43, 103.63 (7Â1C, C-1),
127.02, 127.09, 127.27, 127.31, 127.46, 127.56, 127.60,
127.71, 127.75, 127.83, 127.87 (3Â10C, 3Â5C, CH Ph),
137.54, 137.69, 137.81, 137.93, 138.16 (3Â2C, 3Â1C, C
Ph), 168.61, 168.70, 168.84, 169.38, 169.73 (1Â3C, 3Â2C,
3Â1C, COOtBu).
Mixture 21 (2 compounds). 87%; Rf=0.4±0.5 (H:E,
4:1); 13C NMR: d 28.10, 28.19, 28.66 (2Â9C, OtBu),
59.39, 63.46 (2Â1C, C-5), 61.20, 61.33 (2Â1C, C-7), 68.83,
68.97, 69.45, 69.79, 69.99, 70.62, 70.69, 71.79 (2Â4C, C-6,
CH2COOtBu), 77.74, 78.75, 80.16, 82.71, 84.83 (2Â3C, C-
2, C-3, C-4), 81.02, 81.38, 81.54 (2Â3C, OtBu), 97.46,
103.23 (2Â1C, C-1), 169.13, 169.33, 169.49, 169.58,
169.67 (2Â3C, COOtBu).
Mixture 22 (6 compounds). 87%; Rf=0.4±0.5 (H:E,
4:1); 13C NMR: d 28.11 (6Â6C, OtBu), 59.74, 59.88, 59.99,
63.66, 63.84, 63.92 (6Â1C, C-5), 61.25, 61.39, 61.54, 61.64
(6Â1C, C-7), 68.46, 69.26, 69.73, 69.89, 70.02, 70.39,
70.97, 71.08, 71.89, 73.04, 73.16, 73.24, 74.58, 75.39, 75.51
(6Â4C, C-6, CH2Ph, CH2COOtBu), 77.14, 78.19, 78.48,
78.67, 79.05, 79.43, 79.67, 80.19, 81.01, 81.41, 81.87,
Mixture 26 (2 compounds). 87%; Rf=0.4±0.55 (H:E,
4:1); 13C NMR: d 27.99 (2Â9C, OtBu), 29.64, 29.99
(2Â1C, C-7), 60.02, 63.96 (2Â1C, C-5), 67.96, 69.18,
69.39, 69.43, 69.55, 69.94, 70.90, 71.00 (2Â4C, C-6, CH2
COOtBu), 77.45, 78.86, 80.02, 82.51, 82.98, 84.86 (2Â3C,
C-2, C-3, C-4), 81.05, 81.19, 81.39, 81.44, 81.48 (2Â3C,

520
N. Moitessier et al. / Bioorg. Med. Chem. 9 (2001) 511±523
OtBu), 97.69, 103.49 (2Â1C, C-1), 169.13, 169.23,
169.32, 169.56, 169.80, 169.82 (2Â3C, COOtBu).
Mixture 34 (2 compounds). 95%; 13C NMR: d 10.17
(2Â1C, CH3), 19.03, 19.09 (2Â1C, CH2CH3), 46.83, 46.89
(2Â1C, C-7), 49.23, 49.28 (2Â1C, CH2N), 58.94, 62.44,
62.69, 64.61 (2Â2C, C-5, C-6), 67.88, 68.10, 68.65, 69.26,
69.65, 69.78 (2Â3C, CH2COOH), 77.71, 78.05, 80.04,
81.58, 81.72, 83.44 (2Â3C, C-2, C-3, C-4), 96.40, 102.10
(2Â1C, C-1), 116.90 (q, JC,F 290 Hz, CF3), 163.24 (q,
JC,F 35 Hz, COCF3), 173.78, 174.14, 174.19, 174.29,
174.47 (2Â3C, COOH).
Mixture 35 (6 compounds). 94%; 13C NMR: d 10.80,
10.84 (6Â1C, CH3), 19.72 (6Â1C, CH2CH3), 47.51
(6Â1C, C-7), 49.86, 49.94 (6Â1C, CH2N), 59.68, 59.89,
63.02, 63.27, 65.29 (6Â2C, C-5, C-6), 68.41, 68.57, 68.90,
69.46, 69.54, 69.89, 70.10, 70.18, 70.35, 73.48, 73.71, 74.25,
74.59, 75.11, 75.65, 75.98 (6Â3C, CH2 COOH, CH2Ph),
77.26, 77.67, 78.29, 78.47, 78.95, 79.61, 80.54, 80.70, 80.93,
81.01, 81.98, 82.23, 82.46, 83.65, 83.82 (6Â3C, C-2, C-3,
C-4), 96.92, 97.21, 97.39, 102.76, 103.17 (6Â1C, C-1),
116.97 (q, JC,F 290 Hz, CF3), 129.04, 129.14, 129.41,
129.53 (6Â5C, CH Ph), 137.40, 138.87, 138.15 (6Â1C, C
Ph), 162.91 (q, JC,F 35 Hz, COCF3), 168.57, 168.68,
168.81, 168.90, 169.06, 169.17, 169.29, 169.43 (6Â2C,
COOH).
Mixture 27 (6 compounds). 89%; Rf=0.4±0.55 (H:E,
4:1); 13C NMR: d 27.90 (6Â6C, OtBu), 29.61, 29.91, 30.05
(6Â1C, C-7), 59.94, 60.07, 60.18, 63.73, 63.82, 63.92
(6Â1C, C-5), 67.67, 67.92, 68.30, 69.22, 69.36, 69.63, 70.07,
70.94, 71.05, 73.06, 73.30, 74.48, 75.35, 75.47 (6Â4C, C-6,
CH2Ph, CH2COOtBu), 77.83, 77.95, 78.06, 78.17, 79.21,
79.41, 79.59, 80.47, 81.25, 81.57, 82.40, 82.56, 82.73,
83.19, 84.46, 84.99 (6Â3C, C-2, C-3, C-4), 81.08, 81.45
(6Â2C, OtBu), 96.84, 97.85, 103.42, 103.54, 104.79 (6Â1C,
C-1), 127.45, 127.57, 127.68, 127.84, 127.92, 128.00,
128.11, 128.23 (6Â5C, CH Ph), 138.00, 138.07, 138.18,
138.32 (6Â1C, C Ph), 168.98, 169.11, 169.21, 169.38,
169.62, 169.75, 169.89 (6Â2C, COOtBu).
Mixture 28 (6 compounds). 89%; Rf=0.4±0.55 (H:E,
4:1); 13C NMR: d 27.94 (6Â3C, OtBu), 29.68, 30.02, 30.16
(6Â1C, C-7), 60.12, 60.21, 60.28, 63.73, 63.84, 63.98
(6Â1C, C-5), 67.76, 68.00, 69.27, 69.43, 69.80, 70.27,
71.03, 71.16, 73.19, 73.33, 74.68, 75.44, 75.55 (6Â4C, C-6,
CH2Ph, CH2COOtBu), 77.10, 77.28, 78.11, 78.74, 78.87,
79.19, 79.57, 80.44, 81.21, 81.34, 81.39, 81.56, 82.32,
82.83, 83.00, 83.68, 84.76 (6Â3C, C-2, C-3, C-4), 81.11,
81.48 (6Â1C, OtBu), 97.04, 97.12, 98.04, 103.62, 103.87,
104.05 (6Â1C, C-1), 127.51, 127.66, 127.89, 128.00, 128.27,
129.68 (6Â10C, CH Ph), 137.99, 138.25, 138.35, 138.58
(6Â2C, C Ph), 168.99, 169.58, 169.78 (6Â1C, COOtBu).
Mixture 36 (6 compounds). 93%; 13C NMR: d 10.61
(6Â1C, CH3), 19.57, 19.65 (6Â1C, CH2CH3), 47.14
(6Â1C, C-7), 49.52 (6Â1C, CH2N ), 60.02, 63.13, 63.17,
63.34, 63.48, 63.57, 64.99, 65.13 (6Â2C, C-5, C-6),
68.45, 68.84, 69.59, 69.84, 70.04, 70.29, 73.18, 73.41,
73.53, 74.05, 74.24, 74.95, 75.51, 75.74 (6Â3C, CH2
COOH, CH2Ph), 77.33, 77.60, 77.84, 78.15, 78.81,
79.10, 79.39, 79.92, 80.88, 81.13, 81.33, 81.81, 82.75,
83.18, 83.91 (6Â3C, C-2, C-3, C-4), 97.18, 97.69,
103.01, 103.46, 103.63 (6Â1C, C-1), 117.01 (q, JC,F 290
Hz, CF3), 128.32, 128.40, 128.51, 128.86, 129.06
(6Â10C, CH Ph), 137.55, 138.16, 138.28, 138.47, 138.59,
138.70 (6Â2C, C Ph), 162.20 (q, JC,F 35 Hz, COCF3),
173.27, 173.48, 173.56, 173.64, 173.76, 174.16 (6Â1C,
COOH).
Libraries 30 and 31 were not accurately characterized
due to extensive signal overlap. The following sub-
libraries 32±36 prepared for the deconvolution process
of the main library 31a have been characterized. Their
preparation involved amination±deprotection of the
sub-libraries 24±28 according to the above general
procedures.
Mixture 32 (7 compounds). 90%; 13C NMR: d 10.50
(7Â1C, CH3), 19.41 (7Â1C, CH2CH3), 47.08 (7Â1C, C-
7), 49.55ꢀ(7Â1C, CH2N), 59.35, 59.66, 63.02ꢀ(7Â2C, C-
5, C-6), 68.07, 68.24, 68.41, 68.53, 68.72, 68.89, 69.21,
69.97, 73.46, 74.04, 75.67 (7Â3C, CH2COOH, CH2Ph),
77.30, 78.02, 78.27, 78.54, 78.68, 79.36, 80.19, 80.35,
80.56, 80.78, 80.97, 81.59, 81.71ꢀ(7Â3C, C-2, C-3, C-4),
96.77, 97.18 (7Â1C, C-1), 116.67 (q, JC,F 290 Hz, CF3),
128.95, 129.14 (3Â5C, 3Â10C, CH Ph), 137.14, 137.39,
137.58, 137.84 (3Â1C, 3Â2C, C Ph), 163.16 (q, JC,F 35 Hz,
COCF3), 173.95, 174.34, 174.48, 174.66 (3Â1C, 3Â2C,
1Â3C, COOH).
Deconvolution process: ®nal steps and resynthesis of the
®nal compound 39 according to general procedures
Allyl 2,3-di-O-benzyl-4-O-(tert-butoxycarbonylmethyl)-ꢀ-
D-xylopyranoside (41). Allyl 2,3-di-O-benzyl-a-d-xylo-
pyranoside (40) (801 mg, 2.16 mmol) was treated with
NaH (346 mg, 8.65 mmol) and tert-butylbromoacetate
(1.28 mL) to provide 41 (1.01 g, 2.09 mmol, 97%) as a
colorless oil; Rf=0.52 (H:E, 4:1); [a]D +39.5 (c 1.0,
1
CHCl3); IR: 1747 cmÀ1; H NMR: d 1.48 (s, 9H, tBu),
Mixture 33 (7 compounds). 94%; 13C NMR: d 9.97
(7Â1C, CH3), 18.86 (7Â1C, CH2CH3), 46.69 (7Â1C, C-
7), 48.77, 48.98 (7Â1C, CH2N), 62.26, 62.49, 62.84,
64.49, 64.82 (7Â2C, C-5, C-6), 67.57, 69.03, 69.18, 69.42,
72.79, 72.89, 74.37, 74.87 (7Â3C, CH2COOH, CH2Ph),
76.40, 76.75, 77.55, 77.88, 78.13, 78.27, 79.82, 80.21, 80.39,
81.41, 81.55, 81.90, 82.09, 82.73, 82.97 (7Â3C, C-2, C-3,
C-4), 101.97, 102.16, 102.36, 102.55 (7Â1C, C-1), 116.16
(q, JC,F 290 Hz, CF3), 128.28, 128.48, 128.64 (3Â5C,
3Â10C, CH Ph), 137.00, 137.27 (3Â1C, 3Â2C, C Ph),
162.62 (q, JC,F 35 Hz, COCF3), 173.82, 174.13, 174.48
(3Â1C, 3Â2C, 1Â3C, COOH).
3.47 (m, 2H, H-2, H-5), 3.60 (dd, 1H, J2,3 10.5, J3,4 10.5
0
Hz, H-3), 3.82 (ddd, 1H, J4,5 4, J4,5 2 Hz, H-4), 3.92
(dd, J5,5 9.5 Hz, H-50), 3.98 (ddd, 1H, J6,6 13, J6,7 7, J6,8
0
0
1.5 Hz, H-6), 4.18 (dd, 1H, J6 ,7 5 Hz, J6 ,8 1 Hz, H-60),
4.18 (s, 2H, CH2COOtBu), 4.63 (d, 1H, Jgem 12 Hz,
CH2Ph), 4.73 (d, 1H, J1,2 4 Hz, H-1), 4.76 (d, 1H,
CH2Ph), 4.82 (d, 1H, Jgem 11 Hz, CH2Ph), 4.93 (d, 1H,
0
0
0
CH2Ph), 5.23 (dd, 1H, J7,8 10.5, J8,8 1.5 Hz, H-8), 5.33
0
(dd, 1H, J7,8 17 Hz, H-80), 5.94 (m, 1H, H-7), 7.34 (m,
10H, Ph); 13C NMR: d 27.9 (3C, t-Bu), 60.0 (1C, C-5),
67.9 (1C, C-6), 69.4 (1C, CH2COOtBu), 73.0, 75.5 (2C,
CH2Ph), 79.0, 79.5 (2C, C-2, C-4), 81.4 (1C, C-3), 81.5

N. Moitessier et al. / Bioorg. Med. Chem. 9 (2001) 511±523
521
(1C, C(CH3)3), 95.4 (1C, C-1), 117.9 (1C, C-8), 127.45,
127.66, 127.85, 127.93, 128.20, 128.24 (10C, CH Ph),
133.6 (1C, C-7), 138.0, 138.6 (2C, C Ph), 169.6 (1C,
COOtBu); EIMS (m/z): 485.0 (M+H)+ꢁ , 426.9, 371.2,
277.2, 187.1, 91.1.
1H, H-1), 4.73 (d, 1H, CH2Ph), 4.81 (d, 1H, Jgem 11 Hz,
CH2Ph), 4.91 (d, 1H, CH2Ph), 7.34 (m, 10H, Ph); 13C
NMR: d 11.1 (1C, CH3), 22.4 (1C, CH2), 27.9 (3C, t-
Bu), 47.5 (1C, C-7), 49.7 (1C, CH2N), 60.0 (1C, C-5),
63.0 (1C, C-6), 69.4 (1C, CH2COOtBu), 74.0, 75.6 (2C,
CH2Ph), 78.9, 79.5 (2C, C-2, C-4), 81.0 (1C, C-3), 82.1
(1C, t-Bu), 96.9 (1C, C-1), 127.82, 127.94, 128.43,
128.59, 128.65 (10C, CH Ph), 137.0, 138.3 (2C, C Phꢁ),
(2-Hydroxy-ethyl) 2,3-di-O-benzyl-4-O-(tert-butoxycarb-
onylmethyl)-ꢀ-D-xylopyranoside (42). Compound 41
(1.00 g, 2.07 mmol) was treated with O3 then sodium
borohydride (312 mg, 8.2 mmol) to yield 42 (995 mg,
2.04 mmol, 98%) as a colorless oil; Rf=0.50 (H:E, 1:4);
169.9 (1C, COOtBu); EIMS (m/z): 530.1 (M+H)+
,
500.6, 474.6, 467.8, 439.8, 427.8, 397.9, 114.1, 91.0.
1
[a]D +41.8 (c 1.0, CHCl3); IR: 3470, 1747 cmÀ1; H
(N-(tert-Butoxycarbonyl)-2-propylamino-ethyl) 2,3-di-O-
benzyl-4-O-(tert-butoxy carbonylmethyl)-ꢀ-D-xylopyr-
anoside (45). Compound 44 (321 mg, 0.61 mmol) was
treated with BOC2O (0.095 mL, 1 mmol) and triethyla-
mine (0.155 mL, 1 mmol) to give 45 (345 mg, 0.55 mmol,
90%); Rf=0.50 (H:E, 4:1); [a]D +50.5 (c 1.8, CHCl3);
NMR: d 1.47 (s, 9H, tBu), 2.71 (m, 1H, OH), 3.47 (m,
2H, H-2, H-5), 3.53 (m, 1H, H-6), 3.62 (m, 1H, H-60),
3.72 (m, 3H, H-7, H-70), 3.83 (dd, J4,5 5, J5,5 10.5 Hz, H-
0
50), 3.92 (dd, 1H, J2,3 10, J3,4 10 Hz, H-3), 4.18 (s, 2H,
CH2COOtBu), 4.65 (d, 1H, Jgem 12 Hz, CH2Ph), 4.73 (d,
1H, J1,2 4 Hz, H-1), 4.75 (d, 1H, CH2Ph), 4.83 (d, 1H,
Jgem 11 Hz, CH2Ph), 4.90 (d, 1H, CH2Ph), 7.34 (m, 10H,
Ph); 13C NMR: d 28.2 (3C, tBu), 60.2 (1C, C-5), 61.6 (1C,
C-7), 68.7 (1C, C-6), 69.6 (1C, CH2COOtBu), 73.5, 75.7
(2C, CH2Ph), 79.3, 79.9 (2C, C-2, C-4), 81.7 (1C, C-3),
81.7 (1C, tBu), 97.6 (1C, C-1), 127.80, 127.95, 128.17,
128.27, 128.54, 128.65 (10C, CH Ph), 138.1, 138.9 (2C,
C Ph), 169.9 (1C, COOtBu); EIMS (m/z): 489.3
(M+H)+ꢁ , 370.9, 277.2, 187.1, 91.1. Anal. calcd for
C27H36O8: C, 66.4; H, 7.4. Found: C, 66.62; H, 7.51.
1
IR: 1748, 1694 cmÀ1; H NMR: d 0.85 (t, 3H, J 7.5 Hz,
CH3), 1.44 (s, 18H, t-Bu), 1.53 (m, 2H, CH2CH3), 3.21
(t, 2H, J 7.5 Hz, CH2N), 3.42 (m, 6H, H-2, H-4, H-5, H-
6, H-7, H-70), 3.81 (m, 3H, H-3, H-50, H-60), 4.18 (s, 2H,
CH2COOtBu), 4.62 (d, 1H, Jgem 12 Hz, CH2Ph), 4.71
(m, 2H, H-1, CH2Ph), 4.81 (d, 1H, Jgem 11 Hz, CH2Ph),
4.91 (d, 1H, CH2Ph), 7.33 (m, 10H, Ph); 13C NMR: d
11.5 (1C, CH3), 21.7, 22.1 (1C, CH2), 28.3, 28.7 (6C,
tBu), 47.0 (1C, C-7), 50.0, 50.5 (1C, CH2NH), 60.3 (1C,
C-5), 66.6 (m, 1C, C-6), 69.7 (1C, CH2 COOtBu), 73.4
(1C, CH2Ph), 75.8 (1C, CH2Ph), 79.2, 80.0 (2C, C-2, C-
4), 79.5, 81.7 (2C, tBu), 81.7 (1C, C-3), 97.3, 97.4 (1C,
C-1), 127.84, 128.05, 128.31, 128.57 (10C, CH Ph),
138.5, 139.0 (2C, C Ph), 155.65 (d, 1C, COOtBu), 169.9
(1C, COOtBu); EIMS (m/z): 630.7 (M+H)+ꢁ , 575.1,
530.5, 474.7, 439.7, 276.9, 186.2, 130.0, 91.1. Anal. calcd
for C35H51NO9: C, 66.7; H, 8.2; N, 2.2. Found: C,
66.61; H, 8.20; N, 2.07.
(2-Bromo-ethyl) 2,3-di-O-benzyl-4-O-(tert-butoxycarbonyl-
methyl)-ꢀ-D-xylopyranoside (43). Compound 42 (550 mg,
1.13 mmol) in DMF (40 mL) was treated with NBS
(599 mg, 3.39 mmol) and PPh3 (889 mg, 3.39 mmol) to give,
after column chromatography, 43 (570mg, 1.04mmol,
92%) as a colorless oil; Rf=0.55 (H:E, 4:1); [a]d +61.3 (c
1
1.1, CHCl3); IR: 1746 cmÀ1; H NMR: d 1.46 (s, 9H, t-
0
Bu), 3.45 (2dd, 2H, J1,2 4, J2,3 10, J4,5 4, J5,5 10 Hz, H-2,
0
H-5), 3.52 (t, 2H, J6,7 6.5 Hz, H-7, H-70), 3.65 (dd, 1H, J4,5
(N-2-Propyl-aminoethyl) 2,3-di-O-benzyl-4-O-(carboxy-
methyl)-ꢀ-D-xylopyranoside (39). Compound 45 (321mg,
0.51mmol) was treated with aqueous tri¯uoroacetic acid
(TFA:H2O, 3:1, 10 mL) to provide, after lyophilization,
tri¯uoroacetate salt 39 (289 mg, 0.51 mmol, 99%) as an
amorphous solid; [a]D +56.8 (c 0.8, CH3OH); 1H NMR
(CD3OD): d 0.96 (t, 3H, J 7.5 Hz, CH3), 1.68 (m, 2H,
CH2CH3), 2.98 (t, 2H, J 7.5 Hz, CH2NH), 3.29 (m, 2H,
H-7, H-70), 3.53 (m, 3H, H-2, H-4, H-5), 3.65 (ddd, 1H,
10 Hz, H-50), 3.80 (m, 1H, H-4), 3.88 (m, 3H, H-3, H-6,
H-60), 4.19 (s, 2H, CH2COOtBu), 4.66 (d, 1H, Jgem 12
Hz, CH2Ph), 4.71 (d, 1H, H-1), 4.78 (d, 1H, CH2Ph),
4.83 (d, 1H, Jgem 11 Hz, CH2Ph), 4.93 (d, 1H, CH2Ph),
7.34 (m, 10H, Ph); 13C NMR: d 28.4 (3C, tBu), 30.2 (1C,
C-7), 60.7 (1C, C-5), 68.2 (1C, C-6), 69.8 (1C,
CH2COOtBu), 73.5, 75.8 (2C, CH2Ph), 79.3, 80.0 (2C, C-
2, C-4), 81.76 (1C, C-3), 81.84 (1C, C(CH3)3), 97.4 (1C, C-
1), 127.92, 128.19, 128.33, 128.65, 128.72 (10C, CH Ph),
137.4, 138.0 (2C, C Ph)ꢁ, 170.0 (1C, COOtBu); EIMS (m/
z): 553.4 (M+2+H)+ , 551.3 (M+H)+ꢁ , 459.1, 461.2,
369.2, 370.9, 263.1, 187.1, 91.1. Anal. calcd for
C27H35O7Br: C, 58.8; H, 6.4. Found C, 58.74; H, 6.26.
0
0
J6,6 11.5, J6,7 5.5, J6,7 5.5 Hz, H-6), 3.85 (m, 2H, H-3,
H-50), 3.98 (ddd, 1H, J6 ,7 5.5, J6` ,7 5.5 Hz, H-60), 4.25
(s, 2H, CH2COOH), 4.66 (s, 2H, CH2Ph), 4.81 (s, 2H,
CH2Ph), 4.91 (d, 1H, J1,2 3.5 Hz, H-1), 7.33 (m, 10H,
Ph); 13C NMR (CD3OD): d 10.6 (1C, CH3), 19.5 (1C,
CH2), 47.3 (1C, C-7), 49.5 (1C, CH2NH), 60.1 (1C, C-
5), 63.0 (m, 1C, C-6), 68.8 (1C, CH2COOH), 74.1, 75.8
(2C, CH2Ph), 79.1, 80.0 (2C, C-2, C-4), 81.2 (1C, C-3),
97.2 (1C, C-1), 116.8 (q, JC,F 290 Hz, CF3), 128.37,
128.52, 128.89, 128.98, 129.08 (10C, CH Ph), 137.6,
138.7 (2C, C Ph), 161.9 (q, JC,F 35 Hz, COCF3) 173.8
(1C, COOH); EIMS (m/z): 474.2 (M+H)+ꢁ , 443.8,
398.7, 381.7, 291.8, 180.7, 91.0.
0
0
0
(2-Propylamino-ethyl) 2,3-di-O-benzyl-4-O-(tert-butoxy-
carbonylmethyl)-ꢀ-D-xylopyranoside (44). Compound 43
(351 mg, 0.64 mmol) in DMF (30 mL) was reacted with
propylamine (3 mL) to a€ord 44 (331 mg, 0.63 mmol,
98%) as a colorless oil; Rf=0.55 (CH2Cl2:MeOH, 9:1);
1
[a]D +58.8 (c 1.0, CHCl3); IR: 3306, 1747 cmÀ1; H
NMR: d 0.89 (t, 3H, J 7.5 Hz, CH3), 1.46 (s, 9H, t-Bu),
1.52 (m, 2H, CH2CH3), 1.90 (m, 1H, NH), 2.57 (t, 2H, J
7.5 Hz, CH2NH), 2.82 (m, 2H, H-7, H-70), 3.42 (dd, 1H,
J1,2 3.5, J2,3 10 Hz, H-2), 3.52 (m, 3H, H-4, H-5, H-6),
3.80 (m, 3H, H-3, H-50, H-60), 4.17 (s, 2H, CH2
COOtBu), 4.62 (d, 1H, Jgem 12 Hz, CH2Ph), 4.71 (d,
Cell±matrix adhesion assay
Assays of cell adhesion to matrix-coated substrata were
performed on bacteriological Petri dishes. 100 mL droplets

522
N. Moitessier et al. / Bioorg. Med. Chem. 9 (2001) 511±523
of a 10 mg/mL solution of either ®bronectin or vitronectin
in phosphate-bu€ered saline (PBS) were deposited on
bacteriological dishes and incubated overnight at 4ꢀC.
After several rinses, the substrata were incubated 30 min
with 3 mg/mL bovine serum albumin (BSA) in PBS
(previously heat-inactivated for 3ꢀmin at 80 ꢀC). The
substrata were thoroughly washed and maintained in
PBS until use. S180 cells were harvested with cell dis-
sociation enzyme-free bu€er (Life Technologies) for
10 min at 37 ꢀC. Cells were pelleted by centrifugation
and incubated for 45 min in DMEM containing 10%
(v/v) FN-depleted FCS. Cells were then centrifuged and
resuspended in DMEM. For adhesion assays, 2.5Â103
cells were deposited on the precoated substrata in
DMEM containing or not the peptidomimetic com-
petitors or RGDS peptide. The dishes were incubated
at 37 ꢀC for 1 h, rinsed with PBS to remove the non-
adherent cells, and ®xed in 5% glutaraldehyde in PBS.
Cells were observed and images were recorded using a
Nikon phase contrast microscope equipped with a high
performance CCD camera. The images were transferred
to a Power Macintosh G3 computer. Cell adhesion was
then quanti®ed by colorimetric analysis. Brie¯y, cells were
stained with 1% crystal violet in 200 mM 2N-morpholino
ethane sulfonic acid (MES) and rinsed in 200 mM MES
bu€er, pH 6. The crystal violet ®xed to the cells was
dissolved in 10% acetic acid and optical density (O.D.)
was measured at 570 nm. The control values correspond
to the O.D. obtained for cells deposited on BSA. 100%
cell adhesion corresponds to the value obtained on FN-
coated substratum at 50 mg/mL. The extent of cell
spreading was quanti®ed using Scion Image software by
measuring the area of adherent cells (at least 30 cells
were measured). At least three independent experiments
were done for each type of substratum (®bronectin or
vitronectin) and with or without competitors.
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Acknowledgements
We thank the Institut Nanceien de Chimie Moleculaire,
the Association pour la Recherche sur le Cancer, for
®nancial support and the Region de Lorraine for a
fellowship to NM. We thank Dr Herve Minoux for
helpful discussions.
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