Diesterification of 3-[(β-cyclodextrinyl)succinamido]propane-1,2-diol catalysed by lipase: Diastereoselectivity or tridimensional substrate specificity?

Article abstract of DOI:10.1002/ejoc.201402414

The transesterification of 3-[(β-cyclodextrinyl)succinamido]propane-1,2-diol with fatty esters catalyzed by immobilized lipase from Mucor miehei occurred with very different conversions of the two diastereoisomers [(R)- or (S)-amidopropanediol]. The highe

Full text of DOI:10.1002/ejoc.201402414

Job/Unit: O42414
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FULL PAPER
DOI: 10.1002/ejoc.201402414
Diesterification of 3-[(β-Cyclodextrinyl)succinamido]propane-1,2-diol Catalysed
by Lipase: Diastereoselectivity or Tridimensional Substrate Specificity?
Cédric Gervaise,[a,b] Véronique Bonnet,*[a] Florian Nolay,[a] Christine Cézard,[a]
Imane Stasik,[a] Catherine Sarazin,[b] and Florence Djedaïni-Pilard[a]
Keywords: Enzyme catalysis / Cyclodextrins / Lipids / Diastereoselectivity / Molecular modeling
The transesterification of 3-[(β-cyclodextrinyl)succinamido]-
propane-1,2-diol with fatty esters catalyzed by immobilized
lipase from Mucor miehei occurred with very different con-
versions of the two diastereoisomers [(R)- or (S)-amido-
propanediol]. The highest conversion observed with the (S)-
amidopropanediol can be related to either lipase diastereo-
selectivity or substrate specificity. To investigate the dia-
stereoselectivity of the lipase, diastereoisomers of the meth-
ylated β-cyclodextrin were replaced by methylated glucopyr-
anoside or methylleucine. No discrimation of the diastereo-
isomers by lipase was observed. Molecular modeling was
performed to assess the lipase selectivity towards the two
diastereoisomers. It was found that the (R)-amidopropanediol
is stabilized by hydrogen bonding with the cyclodextrin rim
resulting in less reactive hydroxy groups.
demonstrated to be more efficient than monocatenary de-
rivatives for the formation of appropriate nanoparticles.[9,10]
One way to control the mono-substitution of permethyl-
ated β-CDs involves chemoenzymatic synthesis. Lipidyl-
cyclodextrins can indeed be obtained by a lipase-catalysed
reaction between mono-substituted permethylated β-CDs
and fatty esters.[10,11] Moreover, it is well known that cyclo-
dextrins interact with enzymes, thereby allowing their pro-
tection or improving their properties.[12]
The successful synthesis of glycerolipidyl-β-cyclodextrins
with fatty esters grafted on to the β-cyclodextrin bearing
a glycerol moiety by lipase-catalysed transesterification has
been reported,[13] and mixtures of diastereoisomers were
observed when racemic glycerol derivatives were used as
substrates.
According to the literature, the transesterification cata-
lysed by lipase may be diastereospecific. This property of
lipase has been shown for hydrolysis reactions[14] as well as
acetylation[15] and transesterification reactions.[16] For ex-
ample, PS lipase from Amano was reported to give a higher
diastereoisomeric excess (de) than AK lipase (62% Ͻ de Ͻ
96%).[16] Finally, Mucor miehei lipase, which was used in
this study, successfully catalysed the synthesis of monoacyl-
glycerol with a diastereoisomeric excess superior to 99%.[17]
As shown in this report, this diastereoselectivity depends
on the enzyme and substrate.
The aim of this study was to understand how lipase from
Mucor miehei acts differently on the two diastereoisomers
of 3-[(β-cyclodextrinyl)succinamido]propane-1,2-diol in the
transesterification with fatty esters. To assess the diastereo-
selectivity or substrate selectivity of the lipase, the reactions
of methyl α-d-glucopyranoside and leucine methyl ester as
Introduction
β-Cyclodextrins (CDs) are cyclic oligosaccharides com-
posed of seven α-d-(1,4)-glucopyranoside moieties. They
are natural compounds produced from starch by the action
of 4-α-glucanotransferases. Their toroidal shape with C-6
primary hydroxy groups on the narrow rim and C-2 and C-
3 secondary hydroxy groups on the wider rim allows them
to encapsulate hydrophobic molecules inside their cavity,
and they have been widely used for many applications.[1,2]
The use of amphiphilic cyclodextrins as mass-transfer addi-
tives in catalysis is very promising.[3,4] In 1999, Hirayama et
al. indicated that hydrophobic CDs obtained by chemical
modifications may serve as slow-release carriers of drugs.[5]
In the last few years, research on the synthesis, properties
and applications of amphiphilic cyclodextrins have been
well documented.[6–8] In particular, these compounds are
promising for drug-delivery purposes. Lipidyl-cyclodextrins,
permethylated β-CDs mono-substituted by fatty esters, are
of special interest for the targeting of plasma membranes.
Mono-substituted compounds, which retain solubility in
aqueous media, with bicatenary fatty acyl chains have been
[a] Université de Picardie Jules Verne, CNRS FRE 3517,
Laboratoire de Glycochimie, des Antimicrobiens et des
Agroressources, Institut de Chimie de Picardie FR 3085 CNRS,
80039 Amiens, France
E-mail: veronique.bonnet@u-picardie.fr
http://www.u-picardie.fr/labo/LG/
[b] Université de Picardie Jules Verne, CNRS FRE 3580, Unité de
Génie Enzymatique et Cellulaire,
80039 Amiens, France
Supporting information for this article is available on the
WWW under http://dx.doi.org/10.1002/ejoc.201402414.
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V. Bonnet et al.
FULL PAPER
analogues of the substrate were studied by molecular mod- verted into the mono- and diesterified compounds 4 and
eling.
5, respectively, whereas more than 80% of 3 [(S)-CD] was
converted into 6 and 7 (Scheme 1). These yields indicate a
clear discrimination between the two diastereoisomers after
24 h in favour of the reaction with 3. Regarding the very
large difference observed in the reactivity of 2 and 3, the
effect of enzyme diastereoselectivity needs to be verified.
As reported previously, the CD cavity interacts with
either enzymes or the acyl chains of the substrates.[10,12] The
same reaction was therefore carried out with the monosac-
charide derivative obtained from methyl α-d-glucopyrano-
side in five steps[20–23] to verify the role of the cyclodextrin
backbone and lipase in this diastereoselectivity.
Results and Discussion
First, the chemoenzymatic synthesis of amphiphilic
cyclodextrin, a mimic of phospholipids, was realized with
racemic (Ϯ)-3-amino-1,2-propanediol, leading to a mixture
of mono- and diesterified diastereoisomers 4–7 in one pot
(see Scheme 1). The mixture could not be separated by
chromatography so the amounts of the diastereoisomers
formed could not be accurately quantified.
Enantiomerically pure (R)- or (S)-3-amino-1,2-propane-
diol were grafted onto 1 to synthesize the diastereomerically
Regioselective tosylation of methyl α-d-glucopyranoside
pure derivatives 2 and 3 separately (Scheme 1), which were with p-toluenesulfonyl chloride in pyridine gave the corre-
then used to study the diastereoselectivity of lipase.
sponding 6-tosyloxy derivative, which, after treatment with
From 6-succinylamido-6-deoxy-permethylated-β-CD (1) NaN3 in DMF, gave the 6-azido-6-deoxy derivative. Subse-
[18]
(Scheme 1), the two diastereoisomers 6I-[(R)-2,3-di- quent methylation of all the hydroxy groups, reduction of
hydroxypropylsuccinamido]-6I-deoxy-permethylated-β-CD the azido group in amine, followed by reaction with succinic
(2) and 6I-[(S)-2,3-dihydroxypropylsuccinamido]-6I-deoxy- anhydride to incorporate the spacer arm led to 8 in an over-
permethylated-β-CD (3) were synthesized in yields of 76 all yield of 21% (Scheme 2).
and 31%, respectively (Scheme 1).
The (R)- and (S)-aminopropanediols were then grafted
Then we performed transesterifications with lipozyme® by amidation leading to compounds 9 and 10, respectively,
and ethyl caprylate under conditions described pre- and lipase-catalysed esterification was carried out to yield
viously.[13,19] Our goal was to obtain bicatenary compounds 11 and 12 from 9, and 13 and 14 from 10. Quantification
that retain their inclusion properties. The reaction was was performed by LC–MS analysis on the crudes in dupli-
stopped after 24 h and the products were carefully purified cate. Compounds 11–14 were then purified and carefully
by silica gel chromatography and characterized by NMR characterized. There was no drastic difference in the conver-
spectroscopy (see the Supporting Information) and mass sion of the lipase-catalysed transesterification with either
spectrometry. A significant difference in reactivity between diastereoisomer (see the Supporting Information) as com-
2 and 3 was observed. Indeed, 25% of 2 [(R)-CD] was con- pared with those achieved with CD derivatives.
Scheme 1. Synthesis of pure diastereoisomers 2 and 3 and their transesterification catalyzed by lipase (yields after purification).
2
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Enzymatic Transesterification
Scheme 2. Synthesis of the glucopyranoside analogue of 3-[(β-cyclodextrinyl)succinamido]propane-1,2-diol (yields after purification).
There was no difference in conversion for the mono- and results obtained with monoacylglycerol.[17] If diastereoselec-
bicatenary compounds. As the two diastereoisomers were tivity is not responsible for the difference in the conversion
converted similarly, it seems that the lipase does not exhibit of 3-[(β-cyclodextrinyl)succinamido]propane-1,2-diol, it can
diastereoselectivity towards this amido derivative.
be hypothesized that the difference is due to the spatial con-
Another model substrate was then used; l-leucine methyl figurations of the R and S substrates. Therefore molecular
ester was chosen for its chirality and amine function to modeling of 2 and 3 was undertaken in an attempt to
mimic the amino-cyclodextrin substrate. The two diastereo- understand the phenomenon.
isomeric leucine derivatives 16 and 17 were subjected to li-
For each diastereoisomer 2 and 3, 50000 snapshots were
pase-catalysed esterification under identical experimental clustered into 30 structures to provide a reliable ensemble of
conditions and quantification showed similar conversions representative conformations. These structures were further
(55–60% conversion; Scheme 3). This last experiment minimized by quantum ab initio calculations at the restric-
clearly demonstrated that lipozyme® is not diastereoselec- ted Hartree–Fock (RHF) level of theory. The results indi-
tive for these transesterification reactions, contrary to the cate that the R configuration (corresponding to dia-
Scheme 3. Reaction with leucine derivatives to explore the effect of cyclodextrin on the reaction (yields after purification).
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FULL PAPER
stereoisomer 2, Figure 1, a) is favoured with a relative en-
ergy of 0 kcalmol–1 compared with 5 kcalmol–1 for 3. This
difference in energy between the most stable structures of
the two configurations represents a significant difference in
stability, as reported previously.[24] Diastereoisomer 2 is
more prone to forming intramolecular hydrogen bonds,
with two of them less than 2 Å in length, the vast majority
of the snapshots showing interactions between the alcohol
groups of the aminopropanediol and the ether functions of
either the methoxy or d-glucose fragments. Consequently,
the arm containing the N,NЈ-dimethylsuccinamide and ami-
nopropanediol moieties is positioned at the side of the
cyclodextrin. The alcohol functions of the S configuration
(corresponding to diastereoisomer 3, see Figure 1, b), in
contrast, are less prone to forming intramolecular hydrogen
bonds. Therefore the OH groups point away from the CD
core and can interact with another substrate. The chirality
of the most stable structures of diastereoisomers 2 and 3
was inverted and new calculations were performed. The re-
sults obtained were not contradictory as the R configura-
tion remains the most stable, even with the initial conditions
favouring the S configuration. This observation was corrob-
orated by the NMR spectroscopic data of these com-
pounds. Indeed, 2 and 3 exhibit different chemical shifts of
the NH amido group (Δδ = 0.11 ppm) and the signal pat-
1
terns in the H NMR and HSQC spectra of the CH2 moie-
ties in the spacer arm (H8 and H9) are very different for 2
and 3, in agreement with the different conformational prop-
erties (Figure 2). Indeed, less hydroxy groups in 2 are avail-
able for nucleophilic attack by the acyl enzyme[25] formed
by the fatty ester and the lipase as a result of the intramo-
lecular hydrogen bonds.
Figure 2. Partial 1H NMR and HSQC spectra of H8 and H9 of
(a) 2 and (b) 3 (10 mm, 600 MHz, 298 K).
Conclusions
The two diastereoisomers of 3-[(β-cyclodextrinyl)suc-
cinamido]propane-1,2-diol, 2 and 3, have been shown to be
very different as substrates for lipase-catalysed transesterifi-
cation. Conversions of 25 and 81%, respectively, were
achieved.
Regarding the reactivity of 3-[(β-cyclodextrinyl)suc-
cinamido]propane-1,2-diol and their analogues, the dia-
stereospecificity of lipozyme®, immobilized Mucor miehei
lipase, was not demonstrated. Indeed, the spacer arm be-
tween the amidoglycerol moiety and the polar head is long
and provides a three-dimensional structure, which is crucial
for lipase catalysis.
The results of molecular modelling ruled out the dia-
stereoselectivity of the lipozyme in the transesterification
of 3-[(β-cyclodextrinyl)succinamido]propane-1,2-diol and
their analogues. The lower conversion obtained with the R
substrate can be explained by the lower availability of the
hydroxy groups on the spacer arm due to the formation of
hydrogen bonds with the CD moiety. The S substrate, which
exhibits no hydrogen bonding with the CD moiety, is more
reactive towards the nucleophilic attack that occurs in the
lipase-catalysed transesterification.
Experimental Section
Figure 1. Lowest-energy structures for (a) diastereoisomer 2 and
(b) diastereoisomer 3 obtained after calculations on 30 different
structures of each compound.
Chemicals: Native β-cyclodextrin was obtained from Wacker
Chemicals (Germany). Fatty esters (99% purity) and Lipozyme®
4
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Enzymatic Transesterification
were from Fluka Chemie GmbH (Germany). Other chemicals were
purchased from Sigma (France), Acros (Belgium), and Fluka
Chemie GmbH (Germany). All the solvents employed for the reac-
tions were distilled once before use. Deuteriated solvents were pur-
chased from Eurisotop (France).
Synthesis of Cyclodextrin Derivatives Used as Acceptors
6I-Succinmonoamido-6I-deoxy-2I,3I-di-O-methylhexakis(2II–VII
,
3II–VII,6II–VII-tri-O-methyl)cyclomaltoheptaose (1): Compound 1
was obtained in five steps from the native β-cyclodextrin, as pre-
viously described in the literature.[18,36] It was purified by column
chromatography on silica gel [CH2Cl2/MeOH (9:1, v/v), Rf = 0.50]
in an overall yield of 25%. Analytical data were identical to those
reported in the literature.[18,36]
Spectroscopic Analysis: Stepwise control of the reactions was read-
ily achieved by ESI-MS in positive-ion mode using a ZQ 4000 qua-
drupole mass spectrometer (Waters-Micromass, Manchester, UK).
High-resolution electrospray mass spectra (ESI-HRMS) in posi-
tive-ion mode were recorded with a Q-TOF Ultima Global instru-
ment (Waters-Micromass, Manchester, UK). Data acquisition and
processing were performed by using MassLynx 4.0 software.
Compound 1 must be freeze-dried for 24 h.
6I-[(R)-2,3-Dihydroxypropylsuccinamido]-6I-deoxy-2I,3I-di-O-meth-
ylhexakis(2II–VII,3II–VII,6II–VII-tri-O-methyl)cyclomaltoheptaose (2):
6I-Succinmonoamido-6I-deoxy-2I,3I-di-O-methylhexakis(2II–VII
,
Structure elucidation of the final products was readily achieved by
3II–VII,6II–VII-tri-O-methyl)cyclomaltoheptaose (1; 100 mg,
0.066 mmol, 1 equiv), 1-hydroxybenzotriazole (89 mg, 0.661 mmol,
10 equiv.) and N,NЈ-diisopropylcarbodiimide (100 μL, 0.661 mmol,
10 equiv.) were dissolved in anhydrous N,N-dimethylformamide
(5 mL) under argon. The reaction mixture was stirred at room tem-
perature for 90 min and then a solution of (R)-3-amino-1,2-pro-
panediol (9 mg, 0.099 mmol, 1.5 equiv. in 5 mL of chloroform and
a few drops of triethylamine) were added. The reaction mixture
was stirred at room temperature overnight. The solvent was then
removed by concentration under reduced pressure and the residue
purified by column chromatography on silica gel [CH2Cl2/MeOH
(98:2, 9:1, 8:2)] to give the product in 76% yield. Rf = 0.30 [CH2Cl2/
MeOH (9:1, v/v)]. [α]2D0 = 120.7 (CHCl3, 0.5 gdL–1). 1H NMR
(600 MHz, CDCl3): δ = 6.67 [t, JNHC7-CH2(6) = 6.1 Hz, 1 H, NHC7],
6.40 (t, JNHC10-11 = 5.6 Hz, 1 H, NHC10), 5.15–5.05 (m, 7 H, 1-
HI–VII), 3.92–3.70 (m, 15 H, 5-HI–VII, 6a-HI–VII, 12-H), 3.70–3.28
(m, 25 H, 3-HI–VII, 4-HI–VII, 6b-HI–VII,11-H, 13-H), 3.64, 3.63, 3.62
(s, 18 H, 6-OCH3), 3.52, 3.51, 3.50, 3.49 (s, 21 H, 3-OCH3), 3.43,
3.40 (s, 21 H, 2-OCH3), 3.23–3.13 (m, 7 H, 2-HI–VII), 2.60–2.43 (m,
4 H, 8-H, 9-H) ppm. 13C NMR (150 MHz, CDCl3): δ = 174.0,
172.5 (2 C, C7, C-10), 99.3, 99.2, 99.1, 99.0, 98.9, 98.7 (7 C, C-
1I–VII), 82.3, 82.1, 81.9, 81.7, 81.6, 81.2, 80.4, 80.2, 79.7 (21 C, C-
2I–VII, C-3I–VII, C-4I–VII), 71.7, 71.4, 71.2 (6 C, C-6II–VII), 71.2, 71.0
(7 C, C-5I–VII), 70.3 (1 C, C-12), 63.9 (1 C, C-13), 61.7, 61.6, 61.5,
61.4 (6 C, 6-OCH3), 59.7, 59.2, 59.1, 58.8, 58.7, 68.6, 58.5 (14 C,
2-OCH3, 3-OCH3), 42.5 (1 C, C-11), 40.4 (1 C, C-6I), 31.8, 31.7
(2 C, C-8, C-9) ppm. HRMS (ESI): calcd. for C69H122N2O38Na
1609.7573 [M + Na]+; found 1609.7603.
1
standard H, 13C, COSY, and HSQC NMR experiments at 600 or
300.13 MHz using a Bruker DRX 600 or AMX 300 spectrometer
1
at 298 K. Me4Si was used as internal standard. H NMR spectro-
scopic spectra were recorded with 16 K data points. Samples were
dissolved in CDCl3. Optical rotations were measured with a Per-
kin–Elmer 343 digital polarimeter using a sodium lamp (λ =
589 nm) at 20 °C.
Chromatographic Separations: Analytical TLC was performed on
Silica Gel 60 F254 plates (Merck, Germany; eluent: CH2Cl2/
MeOH, 9:1) followed by charring with vanillin/H2SO4.
The LC-MS experiments were carried out by using an HPLC linked
to an ESI-ZQ mass spectrometer (Waters-Micromass, Manchester,
UK) with a Prevail C18 column (4.6ϫ 250ϫ 5 μm; rate 1 mL/min)
and water/MeOH as eluent (100% H2O for 10 min then ramped to
100% MeOH for 30 min, then held for 10 min).
Molecular Modelling: Initial geometries of the diastereoisomers 2
and 3 studied in this work were constructed by using the LEaP
program from the AmberTools 1.2 distribution following the strat-
egy and methodology previously established.[26] With the exception
of amino-propanediol, the fragments (i.e., α-d-glucose, methoxy
and N,NЈ-dimethylsuccinamide units) were taken from the
R.E.DD.B. database[27] under project F-85 (http://q4md-force-
fieldtools.org/REDDB/). The amino-propanediol fragment was de-
fined and parametrized according to the strategy developed pre-
viously for the aforementioned fragments by using the RED pro-
gram[28] along with the RED server.[29] To obtain representative
ensembles of conformations for these two diastereoisomers, cluster-
ing of the molecular configurations from molecular dynamics
(MD) trajectories was performed. MD simulations were carried out
by using the SANDER module of the AMBER10 program suite[30]
in both the gas phase and by using a generalized Born implicit
solvent model. Classical MD simulations of 50 ns duration were
performed and the configurations of the systems were stored at
intervals of 1 ps. A distance cut-off of 9.0 Å was applied.
6I-[(S)-2,3-Dihydroxypropylsuccinamido]-6I-deoxy-2I,3I-di-O-meth-
ylhexakis(2II–VII,3II–VII,6II–VII-tri-O-methyl)cyclomaltoheptaose (3):
6I-Succinmonoamido-6I-deoxy-2I,3I-di-O-methylhexakis(2II–VII
,
3II–VII,6II–VII-tri-O-methyl)cyclomaltoheptaose (1; 100 mg,
0.066 mmol, 1 equiv.), 1-hydroxybenzotriazole (89 mg, 0.661 mmol,
10 equiv.) and N,NЈ-diisopropylcarbodiimide (100 μL, 0.661 mmol,
10 equiv.) were dissolved in anhydrous N,N-dimethylformamide
(5 mL) under argon. The reaction mixture was stirred at room tem-
perature for 90 min and then a solution of (S)-3-amino-1,2-pro-
panediol (60 mg, 0.099 mmol, 10 equiv. in 5 mL of chloroform and
a few drops of triethylamine) were added. The reaction mixture was
stirred at room temperature for 24 d. The solvent was removed by
evaporation under reduced pressure and the residue purified by col-
umn chromatography on silica gel [CH2Cl2/MeOH (100:0Ǟ8:2)]
to give the product in 31% yield. Rf = 0.30 [CH2Cl2/MeOH (95:5,
v/v)]. [α]2D0 = +112.6 (CHCl3, 0.5 gdL–1). 1H NMR (600 MHz,
CDCl3): δ = 6.79 (t, JNHC7-CH2(6) = 5.8 Hz, 1 H, NHC7), 6.50 (t,
JNHC10-11 = 5.5 Hz, 1 H, NHC10), 5.17–5.02 (m, 7 H, 1-HI–VII),
3.89–3.73 (m, 14 H, 5-HI–VII, 6a-HII–VII, 12-H), 3.73–3.31 (m, 26,
3-HI–VII, 4-HI–VII, 6b-HI–VII, 6a-HI, 11-H, 13-H), 3.64, 3.63, 3.62
(s, 18 H, 6-OCH3), 3.50, 3.49 (s, 21 H, 3-OCH3), 3.42, 3.38, 3.37
(s, 21 H, 2-OCH3), 3.22–3.10 (m, 7 H, 2-HI–VII), 2.65–2.43 (m, 4
The q4md-CD force field[26] was used to simulate the substituted
CDs: geometrical parameters from AMBER99SB[31] were used to
describe the N,NЈ-dimethylsuccinamide and amino-propanediol
fragments, whereas parameters from GLYCAM04[32–34] were used
to describe the carbohydrate and organic fragments. Scaling factors
for 1–4 non-bonded interactions were set to a value of 1.2 and 2.0
for electrostatic and van der Waals interactions, respectively.
The trajectories were analysed by using the PTRAJ module avail-
able in AmberTools 1.2. The structures obtained from the cluster-
ing of the trajectories were further optimized by means of quantum
chemical ab initio calculations. The Gaussian 09 program (revision
A.2)[35] was used to perform quantum chemical calculations. The
structures were optimized at the restricted Hartree–Fock level of
theory by using the 3-21G basis set.
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H, 8-H, 9-H) ppm. 13C NMR (150 MHz, CDCl3): δ = 174.0, 172.8
(2 C, C-7, C-10), 99.2, 99.1, 99.0, 98.9, 98.7 (7 C, C-1I–VII), 82.2,
82.1, 82.0, 81.9, 81.8, 81.7, 81.6, 81.1, 80.7, 80.2, 79.7 (21 C, C-
6I-[(S)-3-Caproyl-2-hydroxypropylsuccinamido]-6I-deoxy-2I,3I-di-O-
methylhexakis(2II–VII,3II–VII,6II–VII-tri-O-methyl)cyclomaltoheptaose
(6) and 6I-[(S)-2,3-Dicaproylpropylamidosuccinylamido]-6I-deoxy-
2I–VII, C-3I–VII, C-4I–VII), 71.7, 71.5, 71.4 (6 C, C-6II–VII), 71.5, 71.4, 2I,3I-di-O-methylhexakis(2II–VII,3II–VII,6II–VII-tri-O-methyl)cyclo-
71.1, 70.9 (7 C, C-5I–VII), 70.2 (1 C, C-12), 63.9 (1 C, C-13), 61.7,
61.6, 61.5, 61.4 (6 C, 6-OCH3), 59.7, 59.2, 58.8, 58.7, 58.6, 58.5 (14
maltoheptaose (7): 6I-[(S)-2,3-Dihydroxypropylsuccinamido]-6I-de-
oxy-2I,3I-di-O-methylhexakis(2II–VII,3II–VII,6II–VII-tri-O-methyl)-
C, 2-OCH3, 3-OCH3), 42.6 (1 C, C-11), 40.4 (1 C, C-6I), 31.8 (2 C, cyclomaltoheptaose (3; 25 mg, 0.016 mmol, 1 equiv.) and lipo-
C-8, C-9) ppm. HRMS (ESI): calcd. for C69H122N2O38Na
zyme® (15 mg) were added to ethyl caprylate (7.5 mL). The reac-
tion mixture was stirred at 50 °C for 24 h. Then the solution was
filtered and the residue purified by column chromatography on sil-
ica gel [CH2Cl2/MeOH (100:0, 97:3)] to give 6 and 7 in yields of
52 and 29%, respectively.
1609.7573 [M + Na]+; found 1609.7568.
6I-[(R)-3-Caproyl-2-hydroxypropylsuccinamido]-6I-deoxy-2I,3I-di-O-
methylhexakis(2II–VII,3II–VII,6II–VII-tri-O-methyl)cyclomaltoheptaose
(4) and 6I-[(R)-2,3-Dicaproylpropylamidosuccinylamido]-6I-deoxy-
2I,3I-di-O-methylhexakis(2II–VII,3II–VII,6II–VII-tri-O-methyl)cyclo-
maltoheptaose (5): 6I-[(R)-2,3-Dihydroxypropylsuccinamido]-6I-de-
oxy-2I,3I-di-O-methylhexakis(2II–VII,3II–VII,6II–VII-tri-O-methyl)-
cyclomaltoheptaose (2, 25 mg, 0.016 mmol, 1 equiv.) and lipo-
zyme® (15 mg) were added to ethyl caprylate (7.5 mL). The reac-
tion mixture was stirred at 50 °C for 24 h. Then the solution was
filtered and the residue purified by column chromatography on sil-
ica gel to yield the products.
6: Rf = 0.07 [CH2Cl2/MeOH (9:1, v/v)]. [α]2D0 = +40.6 (MeOH,
1
0.5 gdL–1). H NMR (600 MHz, CDCl3): δ = 6.61 (t, JNHC7-CH2(6)
= 5.9 Hz, 1 H, NHC7), 6.37 (t, JNHC10-11 = 5.7 Hz, 1 H, NHC10),
5.16–5.00 (m, 7 H, 1-HI–VII), 4.03 (dd, J12-13a = 5.0, J13a-13b
=
11.3 Hz, 1 H, 13-Ha), 4.00 (dd, J12-13b = 5.7, J13a-13b = 11.3 Hz, 1
H, 13b-H), 3.92–3.26 (m, 30 H, 3-HI–VII, 4-HI–VII, 5-HI–VII, 6-
HI–VII, 11a-H, 12-H), 3.61, 3.59 (s, 18 H, 6-OCH3), 3.47, 3.46, 3.45
(s, 21 H, 3-OCH3), 3.38, 3.35, 3.34 (s, 21 H, 2-OCH3), 3.20–3.08
(m, 8 H, 2-HI–VII, 11b-H), 2.60–2.40 (m, 4 H, 8-H, 9-H), 2.29 (t,
J15-16 = 7.5 Hz, 2 H, 15-H), 1.57 (q, J15-16 = J16-17 = 7.5 Hz, 2 H,
16-H), 1.32–1.12 (m, 8 H, 17-H to 20-H), 0.83 (t, J20-21 = 6.9 Hz,
3 H, 21-H) ppm. 13C NMR (150 MHz, CDCl3): δ = 173.9, 173.5,
172.4 (3 C, C-7, C-10, C-14), 99.1, 99.0, 98.9, 98.8, 98.6 (7 C, C-
1I–VII), 82.2, 82.1, 82.0, 81.9, 81.8, 81.7, 81.6, 81.5, 81.1, 81.0, 80.6,
80.5, 80.2, 80.1, 79.6 (21 C, C-2I–VII, C-3I–VII, C-4I–VII), 71.6, 71.4,
71.3, 71.1, 70.6 (6 C, C-6II–VII), 71.5, 71.2, 71.0 (7 C, C-5I–VII), 68.9
(1 C, C-12), 65.8 (1 C, C-13), 61.6, 61.5, 61.4, 61.3 (6 C, 6-OCH3),
59.6, 59.5, 59.1, 59.0, 58.9, 58.7, 58.6, 58.5, 58.4 (14 C, 2-OCH3, 3-
OCH3), 42.9 (1 C, C-11), 40.2 (1 C, C-6I), 34.1 (1 C, C-15), 31.7,
31.6 (C-8, C-9, CH2), 29.1, 28.9 (CH2), 24.9 (1 C, C-16), 22.6
(CH2 ), 14.1 (1 C, C-21) ppm. HRMS (ESI): calcd. for
C77H136N2O39Na 1735.8618 [M + Na]+; found 1735.8556.
4: CH2Cl2/MeOH (100:0, 95:5, 9:1) as eluent, yield 19%. Rf = 0.13
1
[CH2Cl2/MeOH (95:5 v/v)]. [α]2D0 = +47.3 (MeOH, 0.5 gdL–1). H
NMR (600 MHz, CDCl3): δ = 6.67–6.58 (m, 1 H, NHC7), 6.41–
6.33 (m, 1 H, NHC10), 5.19–5.10 (m, 7 H, 1-HI–VII), 4.08 (dd,
J12-13a = 5.2, J13a-13b = 11.4 Hz, 1 H, 13a-H), 4.04 (dd, J12-13b
=
5.7, J13a-13b = 11.4 Hz, 1 H, 13b-H), 3.95–3.88 (m, 1 H, 12-H),
3.88–3.71 (m, 14 H, 5-HI–VII, 6a-HI–VII), 3.70–3.30 (m, 30 H, 2-
HI–VII, 3-HI–VII, 4-HI–VII, 6b-HI–VII, 11a-H), 3.64, 3.63, 3.62, 3.61
(s, 18 H, 6-OCH3), 3.51, 3.50, 3.49 (s, 21 H, 3-OCH3), 3.42, 3.38,
3.37 (s, 21 H, 2-OCH3), 3.23–3.16 (m, 8 H, 2-HI–VII, 11b-H), 2.53
(t, J8-9 = 4.4 Hz, 4 H, 8-H, 9-H), 2.32 (t, J15-16 = 7.6 Hz, 2 H, 15-
H), (q, J15-16 = J16-17 = 7.6 Hz, 2 H, 16-H), 1.33–1.19 (m, 8 H, 17-
H to 20-H), 0.87 (t, J20-21 = 7.1 Hz, 3 H, 21-H) ppm. 13C NMR
(150 MHz, CDCl3): δ = 174.0, 173.8, 172.4 (3 C, C-7, C-10, C-14),
99.2, 99.1, 98.9, 98.8, 98.7 (7 C, C-1I–VII), 82.2, 82.1, 82.0, 81.9,
81.8, 81.6, 81.5, 81.1, 80.7, 80.5, 80.3, 80.2, 79.7 (21 C, C-2I–VII, C-
3I–VII, C-4I–VII), 71.7, 71.5, 71.3, 71.2, 70.6 (6 C, C-6II–VII), 71.6,
71.1, 71.0, 70.1 (7 C, C-5I–VII), 69.8 (1 C, C-13), 61.7, 61.6, 61.4,
61.3 (6 C, 6-OCH3), 59.6, 59.1, 58.7, 58.6, 58.5, 58.4 (14 C, 2-
OCH3, 3-OCH3), 43.0 (1 C, C-11), 40.3 (1 C, C-6I), 34.2 (1 C, C-
15), 31.8 (C-8, C-9, CH2), 29.8, 29.2, 29.0 (CH2), 25.0 (1 C, C-
16), 22.7 (CH2), 14.2 (1 C, C-21) ppm. HRMS (ESI): calcd. for
C77H136N2O39Na [M + Na]+ 1735.8618; found 1735.8602.
7: Rf = 0.09 [CH2Cl2/MeOH (95:5, v/v)]. [α]2D0 = +65.9 (MeOH,
1
0.5 gdL–1). H NMR (600 MHz, CDCl3): δ = 6.35 (t, JNHC7-CH2(6)
= JNHC10-11 = 5.5 Hz, 2 H, NHC7, NHC10), 5.20–5.05 (m, 8 H, 1-
HI–VII, 12-H), 4.24 (dd, J12-13a = 3.75, J13a-13b = 12.1 Hz, 1 H, 13a-
H), 4.10 (dd, J12-13b = 5.8, J13a-13b = 12.1 Hz, 1 H, 13b-H), 3.90–
3.73 (m, 14 H, 5-HI–VII, 6a-HI–VII), 3.72–3.30 (m, 23 H, 3-HI–VII
,
4-HI–VII, 6b-HI–VII, 11-H), 3.65, 3.64, 3.63 (s, 18 H, 6-OCH3), 3.51,
3.50, 3.49 (s, 21 H, 3-OCH3), 3.43, 3.39, 3.38 (s, 21 H, 2-OCH3),
3.24–3.12 (m, 7 H, 2-HI–VII), 2.50 (s, 4 H, 8-H, 9-H), 2.31, 2.30 (2
t, J15-16 = 7.7 Hz, 4 H, 15-H), 1.75–1.53 (m, 4 H, 16-H), 1.33–1.20
(m, 16 H, 17-H to 20-H), 0.88 (t, J20-21 = 6.5 Hz, 6 H, 21-H) ppm.
13C NMR (150 MHz, CDCl3): δ = 173.0, 171.6 (4 C, C-7, C-10, C-
14), 99.2, 99.1, 99.0, 98.9, 98.7 (7 C, C-1I–VII), 82.3, 82.2, 82.1, 82.0,
81.6, 81.3, 80.7, 80.3, 80.2, 79.7 (21 C, C-2I–VII, C-3I–VII, C-4I–VII),
71.7, 71.5, 71.4, 71.3 (6 C, C-6II–VII), 71.6, 71.1, 70.4 (7 C, C-5I–VII),
70.2 (1 C, C-12), 62.9 (1 C, C-13), 61.7, 61.6, 61.5 (6 C, 6-OCH3),
59.6, 59.2, 59.0, 58.8, 58.7, 58.6, 58.5 (14 C, 2-OCH3, 3-OCH3),
40.4 (1 C, C-6I), 39.9 (1 C, C-11), 34.4, 34.2 (2 C, C-15), 31.8, 31.6
(C-8, C-9, CH2), 29.2, 29.1 (CH2), 25.0 (2 C, C-16), 22.7 (CH2),
14.2 (1 C, C-21) ppm. HRMS (ESI): calcd. for C85H150N2O40Na
1861.9663 [M + Na]+; found 1861.9738.
5: CH2Cl2/MeOH (100:0, 97:3, 95:5) as eluent, yield 6%. Rf = 0.16
[CH2Cl2/MeOH (95:5, v/v)]. [α]2D0 = +40.0 (CHCl3, 0.1 gdL–1). H
1
NMR (600 MHz, CDCl3): δ = 6.40–6.25 (m, 2 H, NHC7, NHC10),
5.20–5.00 (m, 8 H, 1-HI–VII, 12-H), 4.25 (dd, J12-13a = 3.8, J13a-13b
= 12.1 Hz, 1 H, 13a-H), 4.10 (dd, J12-13a = 5.9, J13a-13b = 12.1 Hz,
1 H, 13b-H), 3.92–3.71 (m, 14 H, 5-HI–VII, 6a-HI–VII), 3.71–3.31
(m, 23 H, 3-HI–VII, 4-HI–VII, 6b-HI–VII, 11-H), 3.65, 3.64, 3.63 (s,
18 H, 6-OCH3), 3.51, 3.50 (s, 21 H, 3-OCH3), 3.43, 3.39, 3.38 (s,
21 H, 2-OCH3), 3.23–3.13 (m, 7 H, 2-HI–VII), 2.51, 2.50 (m, 4 H,
8-H, 9-H), 2.31, 2.30 (2 t, J15-16 = 7.7 Hz, 4 H, 15-H), 1.75–1.45
(m, 4 H, 16-H), 1.37–1.20 (m, 16 H, 17-H to 20-H), 0.88 (t, J20-21
= 6.5 Hz, 6 H, 21-H) ppm. 13C NMR (150 MHz, CDCl3): δ = 173.5
(4 C, C-7, C-10, C-14), 99.0, 98.6 (7 C, C-1I–VII), 81.8, 80.9, 80.3
6-Amino-6-deoxy-1,2,3,4-tetra-O-methyl-α-D-glucopyranoside was
(21 C, C-2I–VII, C-3 I–VII, C-4I–VII), 71.2, 70.8, 70.4 (13 C, C-5 I–VII
,
obtained in five steps from methyl-α-d-glucopyranoside as de-
scribed previously in the literature. The analytical data are in agree-
ment with those published.[21]
C-6II–VII), 70.0 (1 C, C-12), 62.6 (1 C, C-13), 61.1, 61.0 (6 C, 6-
OCH3), 59.1, 58.3, 57.1, 56.3 (14 C, 2-OCH3, 3-OCH3), 40.3 (1 C,
C-6I), 39.5 (1 C, C-11), 33.7 (1 C, C-15), 31.3 (2 C, C-8, C-9), 31.7,
39.0, 28.9 (CH2), 24.6 (1 C, C-16), 22.3 (CH2),13.7 (1 C, C-
21) ppm. HRMS (ESI): calcd. for C85H150N2O40Na 1861.9663 [M
+ Na]+; found 1861.9712.
6-Succinmonoamido-6-deoxy-1,2,3,4-tetra-O-methyl-α-D-glucopyr-
anoside (8): Succinic anhydride (2.06 g, 20.6 mmol, 10 equiv.) was
heated at 135 °C. After dissolution, 6-amino-6-deoxy-1,2,3-tri-O-
6
www.eurjoc.org
© 0000 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Eur. J. Org. Chem. 0000, 0–0

Job/Unit: O42414
/KAP1
Date: 19-08-14 17:45:58
Pages: 11
Enzymatic Transesterification
methyl-α-glucopyranoside (483 mg, 2.06 mmol, 1 equiv.) was added
and the reaction mixture was heated at 135 °C for a few minutes.
Then the reaction mixture was dissolved in methanol and the suc-
cinic anhydride was precipitated and was removed by filtration. The
solvent was evaporated under reduced pressure and the residue
purified by column chromatography on silica gel [CH2Cl2/MeOH
[100:0 to 90:10)] to give the product in 93 % yield. Rf = 0.64
H, OMe), 3.40 (s, 3 H, OMe), 3.32 (s, 3 H, OMe), 3.27 (dd, 1 H,
2-H), 3.20 (m, 1 H, 4-H), 2.98 (m, 4 H, 8-H, 9-H) ppm. 13C NMR
(75 MHz, Pyr): δ = 173.92, 172.96 (2 C, CO), 97.98 (1 C, C-1),
84.15 (1 C, C-3), 82.53 (1 C, C-2), 81.70 (1 C, C-4), 72.43 (1 C, C-
12), 70.47 (1 C, C-5), 65.36 (1 C, C-13), 60.83, 60.77, 58.41, 55.22
(4 C, OMe), 43.97 (1 C, C-6), 40.69 (1 C, C-11), 32.37 (2 C, C-8,
C-9) ppm. MS (ESI +): calcd. for C17H32N2O9H 409.2186 [M +
H]+; found 409.2201.
1
[CH2Cl2/MeOH (9:1, v/v)]. H NMR (300 MHz, CDCl3): δ = 8.59
(1 H, COOH), 6.13 (m, 1 H, NH), 4.76 (d, JH1-H2 = 3.56 Hz, 1 H,
1-H), 3.65–3.40 (m, 4 H, 3-H, 5-H, 6-H, 6Ј-H), 3.60, 3.50, 3.49,
3.37 (s, 12 H, OMe), 3.18–3.13 (dd, JH2-H1 = 3.56 Hz, 1 H, 2-H),
2.90 (m, 1 H, 4-H), 2.70 (m, 2 H, 9-H), 2.52 (m, 2 H, 8-H) ppm.
13C NMR (150 MHz, CDCl3): δ = 176.85, 172.08 (2 C, CO), 97.52
(1 C, C-1), 83.48 (1 C, C-3), 81.75 (1 C, C-2), 80.76 (1 C, C-4),
69.07 (1 C, C-5), 60.98–55.37 (4 C, OMe), 39.97 (1 C, C-6), 31.01
(1 C, C-8), 29.77 (1 C, C-9) ppm. MS (ESI +): calcd. for
C14H25NO8Na 358.1478 [M + Na]+; found 358.1493.
6-[(R)-3-Caproyl-2-hydroxypropylsuccinylamido]-6-deoxy-1,2,3,4-
tetra-O-methyl-α-
propylsuccinylamidosuccinylamido]-6-deoxy-1,2,3,4-tetra-O-methyl-
α- -glucopyranoside (12): 6-[(R)-2,3-Dihydroxypropylamido]suc-
D-glucopyranoside (11) and 6-[(R)-2,3-Dicaproyl-
D
cinylamido-6-deoxy-1,2,3,4-tetra-O-methyl-α-d-glucopyranoside
(9; 15 mg, 0.038 mmol, 1 equiv.) and lipozyme® (35 mg) were added
to ethyl caprylate (5 mL). The reaction mixture was stirred at 50 °C
for 23 h. Then the suspension was filtered and the residue purified
by column chromatography on silica gel [CH2Cl2/MeOH (100:0,
95:5, 9:1)] to give 11 and 12 in yields of 36 and 24%, respectively.
6-[(R)-2,3-Dihydroxypropylsuccinamido]-6-deoxy-1,2,3,4-tetra-O-
methyl-α-D-glucopyranoside (9): 6-Succinmonoamido-6-deoxy-
11: Rf = 0.30 [CH2Cl2/MeOH (9:1, v/v)]. 1H NMR (300 MHz, Pyr):
1,2,3-tri-O-methyl-α-glucopyranoside (8; 214 mg, 0.64 mmol,
1 equiv.) and 1-[bis(dimethylamino)methylene]-1H-1,2,3-tria-
zolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU;
355 mg, 0.96 mmol, 1.5 equiv.) were added to anhydrous N,N-di-
methylformamide (4 mL) under argon. The reaction mixture was
stirred at room temperature for 1 h. Then a solution of (R)-3-
amino-1,2-propanediol (583 mg, 6.4 mmol, 10 equiv. in 6 mL of
N,N-dimethylformamide and a few drops of triethylamine) were
added. The reaction mixture was stirred at room temperature for
24 h. The solvent was removed by evaporation under reduced pres-
sure and the residue purified by column chromatography on silica
gel [CH2Cl2/MeOH (98:2 to 9:1)] to give the product in 85% yield.
δ = 9.03 (m, 1 H, NH), 8.68 (m, 1 H, NH), 4.91 (d, JH1-H2
=
3.55 Hz, 1 H, 1-H), 4.53–4.46 (dd, 2 H, 13-H, 13Ј-H), 3.90–3.73
(m, 6 H, 3-H, 5-H, 6-H, 6Ј-H, 11-H), 3.64 (s, 3 H, OMe), 3.62 (s,
3 H, OMe), 3.41 (s, 3 H, OMe), 3.32 (s, 3 H, OMe), 3.28–3.23 (dd,
JH2-H1 = 3.55 Hz, 1 H, 2-H), 3.21 (m, 1 H, 4-H), 2.99 (m, 4 H, 8-
H, 9-H), 2.31 (m, 2 H, 15-H), 1.60 (tt, 2 H, 16-H), 1.27–1.15 (m, 8
H, 17-H to 20-H), 0.82 (t, JH20-H21 = 6.76 Hz, 3 H, 21-H) ppm. 13
C
NMR (75 MHz, Pyr): δ = 174.0, 173.6, 173.0 (3 C, CO), 97.39 (1
C, C-1), 83.58 (1 C, C-3), 81.96 (1 C, C-2), 81.12 (1 C, C-4), 69.92
(1 C, C-12), 68.69 (1 C, C-5), 66.88 (1 C, C-13), 60.28, 60.21, 57.84,
54.63 (4 C, OMe), 43.30 (1 C, C-6), 40.09 (1 C, C-11), 34.07 (1 C,
C-15), 31.73 (2 C, C-8, C-9), 24.97 (1 C, C-16), 32.36, 32.18, 29.64,
23.16 (4 C, C-17 to C-20), 14.55 (1 C, C-21) ppm. MS (ESI +):
calcd. For C25H46N2O10Na 557.3050 [M + Na]+; found 557.3040.
12: Rf = 0.50 [CH2Cl2/MeOH (9:1, v/v)]. 1H NMR (300 MHz, Pyr):
δ = 9.19 (m, 1 H, NH), 8.74 (m, 1 H, NH), 5.66 (m, 1 H, 12-H),
4.91 (d, JH1-H2 = 3.52 Hz, 1 H, 1-H), 4.71–4.49 (dd, J13-H-13-HЈ
3.48 Hz, 2 H, 13-H, 13Ј-H), 3.95–3.73 (m, 6 H, 3-H, 5-H, 6-H, 6Ј-
H, 11-H), 3.64 (s, 3 H, OMe), 3.63 (s, 3 H, OMe), 3.42 (s, 3 H,
OMe), 3.33 (s, 3 H, OMe), 3.28–3.23 (dd, JH2-H1 = 3.52 Hz, 1 H,
2-H), 3.21 (m, 1 H, 4-H), 3.06–2.90 (m, 4 H, 8-H, 9-H), 2.38–2.33
(m, 4 H, 15-H), 1.67–1.58 (tt, 4 H, 16-H), 1.27–1.18 (m, 16 H, 17-
H to 20-H), 0.84 (t, JH20-H21 = 6.68 Hz, 6 H, 21-H) ppm. 13C NMR
(75 MHz, Pyr): δ = 173.7, 173.6, 173.5, 172.9 (4 C, CO), 98.45 (1
C, C-1), 84.64 (1 C, C-3), 82.03 (1 C, C-2), 82.17 (1 C, C-4), 72.00
(1 C, C-12), 70.98 (1 C, C-5), 64.42 (1 C, C-13), 61.35, 61.27, 58.90,
55.68 (4 C, 4 OMe), 41.15 (1 C, C-6), 40.77 (1 C, C-11), 35.22 (2
C, C-15), 32.67 (2 C, C-8, C-9), 26.00 (2 C, C-16), 35.02, 30.07,
30.00, 23.65 (4 C, C-17 to C-20), 15.02 (2 C, C-21) ppm. MS (ESI
+): calcd. for C33H60N2O11Na 683.4095 [M + Na]+; found
683.4067.
1
Rf = 0.38 [CH2Cl2/MeOH (9:1, v/v)]. H NMR (300 MHz, Pyr): δ
= 9.00 (m, 1 H, NH), 8.65 (m, 1 H, NH), 5.92 (2 H, OH), 4.91 (d,
JH1-H2 = 3.61 Hz, 1 H, 1-H), 4.32 (m, 1 H, 12-H), 4.06 (m, 2 H,
13-H), 3.94, 3.81 (m, 2 H, 6-H, 6Ј-H) 3.85 (m, 1 H, 5-H), 3.83 (m,
2 H, 11-H) 3.74 (m, 1 H, 3-H) 3.62 (s, 3 H, OMe), 3.61 (s, 3 H,
OMe), 3.40 (s, 3 H, OMe), 3.32 (s, 3 H, OMe), 3.26 (dd, 1 H, 2-
H), 3.20 (m, 1 H, 4-H), 2.98 (m, 4 H, 8-H, 9-H) ppm. 13C NMR
(75 MHz, Pyr): δ = 173.95, 172.98 (2 C, CO), 97.98 (1 C, C-1),
84.14 (1 C, C-3), 82.54 (1 C, C-2), 81.73 (1 C, C-4), 72.43 (1 C, C-
12), 70.47 (1 C, C-5), 65.35 (1 C, C-13), 60.84, 60.77, 58.42, 55.23
(4 C, OMe), 43.96 (1 C, C-6), 40.71 (1 C, C-11), 32.37 (2 C, C-8,
C-9) ppm. MS (ESI +): calcd. for C17H32N2O9Na 431.2006 [M +
Na]+; found 431.2010.
=
6-[(S)-2,3-Dihydroxypropylsuccinamido]-6-deoxy-1,2,3,4-tetra-O-
methyl-α-D-glucopyranoside (10): 6-Succinmonoamido-6-deoxy-
1,2,3-tri-O-methyl-α-d-glucopyranoside (8; 214 mg, 0.64 mmol,
1 equiv.) and 1-[bis(dimethylamino)methylene]-1H-1,2,3-tri-
azolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU;
355 mg, 0.96 mmol, 1.5 equiv.) were added to anhydrous N,N-di-
methylformamide (4 mL) under argon. The reaction mixture was
stirred at room temperature for 1 h. Then a solution of (S)-3-
amino-1,2-propanediol (583 mg, 6.4 mmol, 10 equiv. in 6 mL of
N,N-dimethylformamide and a few drops of triethylamine) was
added. The reaction mixture was stirred at room temperature for
24 h. The solvent was removed by concentration under reduced
pressure and the residue purified by column chromatography on
silica gel [CH2Cl2/MeOH (98:2 to 9:1)] to give the product in 67%
yield. Rf = 0.38 [CH2Cl2/MeOH (9:1, v/v)]. 1H NMR (300 MHz,
Pyr): δ = 8.99 (m, 1 H, NH), 8.63 (m, 1 H, NH), 5.72 (2 H, OH),
4.91 (d, JH1-H2 = 3.61 Hz, 1 H, 1-H), 4.32 (m, 1 H, 12-H), 4.06 (m,
6-[(S)-3-Caproyl-2-hydroxypropylsuccinamido]-6-deoxy-1,2,3,4-
tetra-O-methyl-α-
D-glucopyranoside (13) and 6-[(S)-2,3-Dicaproyl-
propylamidosuccinylamido]-6-deoxy-1,2,3,4-tetra-O-methyl-α-
D
-
glucopyranoside (14): 6-[(S)-2,3-Dihydroxypropylsuccinamido]-6-
deoxy-1,2,3,4-tetra-O-methyl-α-d-glucopyranoside (10; 15.3 mg,
0.038 mmol, 1 equiv.) and lipozyme® (36 mg) were added to ethyl
caprylate (5.1 mL). The reaction mixture was stirred at 50 °C for
23 h. Then the solution was filtered and the residue purified by
column chromatography on silica gel [CH2Cl2/MeOH (100:0, 95:5,
9:1)] to give 13 and 14 in yields of 36 and 57%, respectively.
2 H, 13-H), 3.96, 3.82 (m, 2 H, 6-H, 6Ј-H) 3.84 (m, 1 H, 5-H), 3.83 13: Rf = 0.28 [CH2Cl2/MeOH (9:1, v/v)]. 1H NMR (300 MHz, Pyr):
(m, 2 H, 11-H) 3.72 (m, 1 H, 3-H), 3.62 (s, 3 H, OMe), 3.61 (s, 3 δ = 8.99 (m, 1 H, NH), 8.64 (m, 1 H, NH), 4.91 (d, 1 H, 1-H),
Eur. J. Org. Chem. 0000, 0–0
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/KAP1
Date: 19-08-14 17:45:58
Pages: 11
V. Bonnet et al.
FULL PAPER
4.53–4.44 (dd, J13-H-13-HЈ = 3.48 Hz, 2 H, 13-H, 13Ј-H), 3.94–3.72
(m, 6 H, 3-H, 5-H, 6-H, 6Ј-H, 11-H), 3.63 (s, 3 H, OMe), 3.62 (s,
3 H, OMe), 3.41 (s, 3 H, OMe), 3.33 (s, 3 H, OMe), 3.27 (dd,
JH2-H1 = 3.49 Hz, 1 H, 2-H), 3.21 (m, 1 H, 4-H), 2.99 (m, 4 H, 8-
H, 9-H), 2.31 (m, 2 H, 15-H), 1.60 (tt, 2 H, 16-H), 1.24–1.12 (m, 8
24.2 (1 C, C-3), 23.2, 21.8 (2 C, C-1, C-2) ppm. HRMS (ESI): calcd.
for C14H26N2O6Na 341.1689 [M + Na]+; found 341.1678.
[(S)-2,3-Dihydroxypropylsuccinamido]-L-leucine Methyl Ester (17):
Succinic anhydride (417 mg, 4.172 mmol, 1.21 equiv.) and l-leucine
methyl ester hydrochloride (500 mg, 3.448 mmol, 1 equiv.) were dis-
solved in distilled dichloromethane (70 mL). Then 4-(dimeth-
ylamino)pyridine (424 mg, 3.483 mmol, 1.01 equiv.) and a few
drops of triethylamine were added. The reaction mixture was
stirred for 17 h at room temperature. Then the reaction was
quenched with a 10% solution of NaHCO3. The aqueous solution
was acidified with 1 n HCl and extracted with ethyl acetate. The
solvent was removed by evaporation under reduced pressure. Then
the obtained product (100 mg, 0.408 mmol, 1 equiv.), 1-hydroxy-
benzotriazole (551 mg, 4.081 mmol, 10 equiv.) and N,NЈ-diisoprop-
ylcarbodiimide (634 μL, 4.081 mmol, 10 equiv.) were dissolved in
anhydrous N,N-dimethylformamide (10 mL) under argon. The re-
action mixture was stirred for 1 h at room temperature. Then a
solution of (S)-3-amino-1,2-propandiol [56 mg in N,N-dimethyl-
formamide (5 mL) and a few drops of triethylamine] was added.
The reaction was stirred for 1 d at room temperature. The solvent
was removed by evaporation under reduced pressure and the resi-
due purified by column chromatography on silica gel [CH2Cl2/
MeOH (100:0 to 8:2)] to give the product in 39% yield over the
two steps. Rf = 0.64 [CH2Cl2/MeOH (8:2, v/v) with Et3N (1%)].
H, 17-H to 20-H), 0.82 (t, JH20-H21 = 6.28 Hz, 3 H, 21-H) ppm. 13
C
NMR (75 MHz, Pyr): δ = 178.54 (3 C, CO), 98.01 (1 C, C-1), 84.18
(1 C, C-3), 82.57 (1 C, C-2), 81.73 (1 C, C-4), 70.51 (1 C, C-12),
69.31 (1 C, C-5), 67.48 (1 C, C-13), 60.88, 60.80, 58.44, 55.24 (4 C,
OMe), 43.92 (1 C, C-6), 40.71 (1 C, C-11), 34.68 (2 C, C-15), 32.36
(2 C, C-8, C-9), 25.57 (2 C, C-16), 32.36, 32.18, 29.64, 23.16 (4
C, C-17 to C-20), 14.55 (2 C, C-21) ppm. MS (ESI +): calcd. for
C25H46N2O10Na 557.3050 [M + Na]+; found 557.3056.
14: Rf = 0.46 [CH2Cl2/MeOH (9:1, v/v)]. 1H NMR (300 MHz, Pyr):
δ = 9.15 (m, 1 H, NH), 8.75 (m, 1 H, NH), 5.65 (m, 1 H, 12-H),
4.93 (d, 1 H, 1-H), 4.68–4.44 (dd, J13-H-13-HЈ = 3.48 Hz, 2 H, 13-
H,13Ј-H), 3.95–3.73 (m, 6 H, 3-H, 5-H, 6-H, 6Ј-H, 11-H), 3.64 (s,
3 H, OMe), 3.63 (s, 3 H, OMe), 3.42 (s, 3 H, OMe), 3.33 (s, 3 H,
OMe), 3.27 (dd, JH2-H1 = 3.67 Hz, 1 H, 2-H), 3.20 (m, 1 H, 4-H),
2.98 (m, 4 H, 8-H, 9-H), 2.34 (m, 4 H, 15-H), 1.63 (tt, 4 H, 16-H),
1.33–1.19 (m, 16 H, 17-H to 20-H), 0.84 (t, JH20-H21 = 6.85 Hz, 6
H, 21-H) ppm. 13C NMR (75 MHz, Pyr): δ = 179.3 (4 C, CO),98.02
(1 C, C-1), 84.19 (1 C, C-3), 82.60 (1 C, C-2), 81.78 (1 C, C-4),
71.59 (1 C, C-12), 70.53 (1 C, C-5), 63.97 (1 C, C-13), 60.89, 60.79,
58.44, 55.24 (4 C, OMe), 40.74 (1 C, C-6), 40.32 (1 C, C-11), 34.77
(2 C, C-15), 32.21 (2 C, C-8, C-9), 25.55 (2 C, C-16), 34.57, 30.36,
29.62, 23.19 (4 C, C-17 to C-20), 14.56 (2 C, C-21) ppm. MS (ESI
+): calcd. for C33H60N2O11Na 683.4095 [M + Na]+; found
683.4086.
[α]2D0 = –37.1 (MeOH, 1.0 gdL–1). H NMR (600 MHz, CDCl3): δ
1
= 8.20 (d, J5-NHC8 = 7.7 Hz, 1 H, NHC8), 7.78 (t, J12-NHC11
=
5.7 Hz, 1 H, NHC11), 4.68 (br., 1 H, OH), 4.48 (br., 1 H, OH),
4.26 (ddd, J4a-5 = 13.1, J5-NHC8 = 7.7, J4b-5 = 5.5 Hz, 1 H, 5-H),
3.61 (s, 3 H, 7-H), 3.56–3.40 (m, 1 H, 13-H), 3.29–3.22 (m, 2 H,
14-H), 3.17 (ddd, J12a-12b = 13.5, J12a-NHC11 = 5.7, J12a-13 = 5.0 Hz,
[(R)-2,3-Dihydroxypropylsuccinamido]-L-leucine Methyl Ester (16):
1 H, 12a-H), 2.95 (ddd, J12a-12b = 13.5, J12b-NHC11 = 5.7, J12b-13
=
Succinic anhydride (417 mg, 4.172 mmol, 1.21 equiv.) and l-leucine
methyl ester hydrochloride (500 mg, 3.448 mmol, 1 equiv.) were dis-
solved in distilled dichloromethane (70 mL). Then 4-(dimeth-
ylamino)pyridine (424 mg, 3.483 mmol, 1.01 equiv.) and a few
drops of triethylamine were added. The reaction mixture was
stirred for 17 h at room temperature. Then the reaction was
quenched with a 10% solution of NaHCO3. The aqueous solution
was acidified with 1 n HCl and extracted with ethyl acetate. The
solvent was removed by evaporation under reduced pressure. Then
the obtained product (100 mg, 0.408 mmol, 1 equiv.), 1-hydroxy-
benzotriazole (551 mg, 4.081 mmol, 10 equiv.) and N,NЈ-diisoprop-
ylcarbodiimide (634 μL, 4.081 mmol, 10 equiv.) were dissolved in
anhydrous N,N-dimethylformamide (10 mL) under argon. The re-
action mixture was stirred for 1 h at room temperature. Then a
solution of (R)-3-amino-1,2-propandiol [56 mg in N,N-dimethyl-
formamide (5 mL) and a few drops of triethylamine] was added.
The reaction was stirred for 1 d at room temperature. The solvent
was removed by evaporation under reduced pressure and the resi-
due purified by column chromatography on silica gel [CH2Cl2/
MeOH (100:0 to 8:2)] to give the product in 41% yield for the two
6.6 Hz, 1 H, 12b-H), 2.45–2.25 (m, 4 H, 9-H, 10-H), 1.70–1.40 (m,
3 H, 3-H, 4-H), 0.88, 0.83 (2 d, J = 6.6, J = 6.4 Hz, 6 H, 1-H, 2-
H) ppm. 13C NMR (150 MHz, CDCl3): δ = 173.1, 171.6 (3 C, C-
6, C-8, C-11), 70.5 (1 C, C-13), 63.6 (1 C, C-14), 51.7 (1 C, C-7),
50.2 (1 C, C-5), 42.2 (1 C, C-12), 38.8 (1 C, C-4), 30.7, 30.5 (2 C,
C-9, C-10), 24.2 (1 C, C-3), 22.7, 21.3 (2 C, C-1, C-2) ppm. HRMS
(ESI): calcd. for C14H26N2O6Na 341.1689 [M + Na]+; found
341.1678.
[(R)-3-Caproyl-2-hydroxypropylsuccinamido]-L-leucine Methyl Ester
(18) and [(R)-2,3-Dicaproylpropylamidosuccinylamido]-
L-leucine
Methyl Ester (19): [(R)-2,3-Dihydroxypropylsuccinamido]-l-leucine
methyl ester (16; 10 mg, 0,031 mmol, 1 equiv.) and lipozyme® were
added to ethyl caprylate (3 mL). The reaction mixture was stirred
for 25 h at 50 °C. Then the solution was filtered and the residue
purified by column chromatography on silica gel [CH2Cl2/MeOH
(100:0 to 8:2)] to afford 18 and 19 in yields of 49 and 13%, respec-
tively.
18: Rf = 0.45 [CH2Cl2/MeOH (95:5, v/v) with Et3N (1%)]. [α]2D0
=
1
steps. Rf = 0.64 [CH2Cl2/MeOH (8:2, v/v) with Et3N (1%)]. [α]2D0
=
–10.0 (MeOH, 0.5 gdL–1). H NMR (600 MHz, CDCl3): δ = 6.47
(t, J12-NHC11 = 5.7 Hz, 1 H, NHC11), 6.29 (d, J5-NHC8 = 7.9 Hz, 1
H, NHC8), 4.61–4.45 (m, 1 H, 5-H), 4.12 (dd, J14a-14b = 11.5,
1
–19.8 (MeOH, 1.0 gdL–1). H NMR (300 MHz, DMSO): δ = 8.20
(d, J5-NHC8 = 7.7 Hz, 1 H, NHC8), 7.78 (t, J12-NHC11 = 5.8 Hz, 1
H, NHC11), 4.68 (br., 1 H, OH), 4.48 (br., 1 H, OH), 4.26 (ddd, J13-14a = 4.9 Hz, 1 H, 14a-H), 4.06 (dd, J14a-14b = 11.5, J13-14b
=
J4a-5 = 13.3, J5-NHC8 = 7.7, J4b-5 = 5.6 Hz, 1 H, 5-H), 3.61 (s, 3 H,
7-H), 3.58–3.40 (m, 1 H, 13-H), 3.28–3.23 (m, 2 H, 14-H), 3.17
(ddd, J12a-12b = 13.5, J12a-NHC11 = 5.8, J12a-13 = 5.3 Hz, 1 H, 12a-
5.9 Hz, 1 H, 14b-H), 3.99–3.86 (m, 1 H, 13-H), 3.73 (s, 3 H, 7-H),
3.51–3.44 (m, 1 H, 12a-H), 3.24 (ddd, J12a-12b = 13.9, J12b-13 = 7.1,
J12b-NHC11 = 5.7 Hz, 1 H, 12b-H), 2.75–2.40 (m, 4 H, 9-H, 10-H),
H), 2.95 (ddd, J12a-12b = 13.5, J12b-13 = 6.6, J12b-NHC11 = 5.8 Hz, 1 2.36–2.25 (m, 2 H, 16-H), 1.72–1.45 (m, 5 H, 3-H, 4-H, 17-H),
H, 12b-H), 2.42–2.23 (m, 4 H, 9-H, 10-H), 1.71–1.40 (m, 3 H, 3- 1.35–1.15 (m, 8 H, 18-H to 21-H), 0.93 (2 d, J = 6.1, J = 5.8 Hz,
H, 4-H), 0.88, 0.83 (2 d, J = 6.6, J = 6.4 Hz, 6 H, 1-H, 2-H) ppm.
13C NMR (75 MHz, DMSO): δ = 173.6, 172.1 (3 C, C-6, C-8, C-
11), 71.0 (1 C, C-13), 64.1 (1 C, C-14), 52.2 (1 C, C-7), 50.7 (1 C,
6 H, 1-H, 2-H), 0.88 (t, J21-22 = 6.9 Hz, 3 H, 22-H) ppm. 13C NMR
(150 MHz, CDCl3): δ = 174.2, 173.7, 172.3 (4 C, C-6, C-8, C-11,
C-15), 69.3 (1 C, C-13), 65.8 (1 C, C-14), 52.5 (1 C, C-7), 51.1 (1
C-5), 42.6 (1 C, C-14), 39.8 (1 C, C-4), 31.1, 31.0 (2 C, C-9, C-10), C, C-5), 43.0 (1 C, C-12), 41.4 (1 C, C-4), 34.3 (1 C, C-16), 31.9,
8
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Eur. J. Org. Chem. 0000, 0–0

Job/Unit: O42414
/KAP1
Date: 19-08-14 17:45:58
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Enzymatic Transesterification
31.8, 31.7 (3 C, C-9, C-10, CH2), 29.2, 29.1, 25.1, 22.7 (4 C, CH2), (2 C, C-1, C-2), 14.2 (1 C, C-22) ppm. HRMS (ESI): calcd. for
22.9 (1 C, C-3), 22.0 (2 C, C-1, C-2), 14.2 (1 C, C-22) ppm. HRMS
(ESI): calcd. for C22H40N2O7Na 467.2733 [M + Na]+; found
467.2732.
C30H54N2O8Na 593.3778 [M + Na]+; found 593.3784.
Supporting Information (see footnote on the first page of this arti-
cle): NMR spectra of all final compounds and chromatograms.
19: Rf = 0.52 [CH2Cl2/MeOH (95:5, v/v) with Et3N (1%)]. [α]2D0
=
0.0 (MeOH, 0.1 gdL–1). H NMR (600 MHz, CDCl3): δ = 6.28 (t,
J12-NHC11 = 5.9 Hz, 1 H, NHC11), 6.25 (d, J5-NHC8 = 8.4 Hz, 1 H,
NHC8), 5.10 (tdd, J12-13 = 6.0, J13-14b = 5.8, J13-14a = 4.1 Hz, 1 H,
13-H), 4.57 (td, J5-NHC8 = 8.4, J4-5 = 5.0 Hz, 1 H, 5-H), 4.25 (dd,
J14a-14b = 12, J13-14a = 4.1 Hz, 1 H, 14a-H), 4.10 (dd, J14a-14b = 12,
1
Acknowledgments
The authors acknowledge Dr. J. M. Garcia Fernandez for helpful
discussions. This work was supported by Le Conseil Régional de
Picardie (grant to C. G.) and by Fonds Européens de Développe-
ment Régional (FEDER) under the scientific “Nanovecteurs” pro-
gram.
J13-14b = 5.8 Hz, 1 H, 14b-H), 3.73 (s, 3 H, 7-H), 3.47 (t, J12-13
=
J12-NHC11 = 6.0 Hz, 1 H, 12-H), 2.57–2.49 (m, 4 H, 9-H, 10-H),
2.33, 2.31 (2 t, J16-17 = 7.6 Hz, 4 H, 16-H), 1.75–1.47 (m, 7 H, 3-
H, 4-H, 17-H), 1.40–1.15 (m, 16 H, 18-H to 21-H), 0.94, 0.93 (2 d,
J = 6.3, J = 6.1 Hz, 6 H, 1-H, 2-H), 0.88 (2 t, J21-22 = 7.1 Hz, 6 H,
22-H) ppm. 13C NMR (150 MHz, CDCl3): δ = 173.6, 173.5, 172.4,
172.2 (5 C, C-6, C-8, C-11, C-15), 70.4 (1 C, C-13), 62.9 (1 C, C-
14), 52.4 (1 C, C-7), 51.1 (1 C, C-5), 41.5 (1 C, C-4), 40.0 (1 C, C-
12), 34.4, 34.2 (2 C, C-16), 31.8, 31.6 (C-9, C-10, CH2), 29.2, 29.1,
25.0, 22.8 (CH2), 22.9 (1 C, C-3), 22.1 (2 C, C-1, C-2), 14.2 (2 C,
C-22) ppm. HRMS (ESI): calcd. for C30H54N2O8Na 593.3778 [M
+ Na]+; found 593.3773.
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Wiley-VCH, Weinheim, Germany, p. 489.
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(20) and [(S)-2,3-Dicaproylpropylamidosuccinylamido]-
L-leucine
Methyl Ester (21): [(S)-2,3-Dihydroxypropylsuccinamido]-l-leucine
methyl ester (17; 10 mg, 0,031 mmol, 1 equiv.) and lipozyme® were
added to ethyl caprylate (3 mL). The reaction mixture was stirred
for 25 h at 50 °C. Then the solution was filtered and the residue
purified by column chromatography on silica gel [CH2Cl2/MeOH
(100:0 to 8:2)] to give 20 and 21 in yields of 26 and 28%, respec-
tively.
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Phenom. Macrocycl. Chem. 2007, 57, 15–20.
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20: Rf = 0.54 [CH2Cl2/MeOH (95:5, v/v) with Et3N (1%)]. [α]2D0
=
1
–14.6 (MeOH, 0.5 gdL–1). H NMR (600MHz, CDCl3): δ = 6.48–
6.40 (m, 1 H, NHC11), 6.33–6.23 (m, 1 H, NHC8), 4.62–4.46 (m,
1 H, 5-H), 4.12–4.02 (m, 2 H, 14-H), 4.02–3.91 (m, 1 H, 13-H),
3.73 (s, 3 H, 7-H), 3.65–3.33 (m, 1 H, 12a-H), 3.16 (ddd, J12a-12b
=
12.9, J12b-13 = 7.4, J12a-NHC11 = 5.4 Hz, 1 H, 12b-H), 2.77–2.40 (m,
4 H, 9-H, 10-H), 2.37–2.22 (m, 2 H, 16-H), 1.70–1.43 (m, 5 H, 3-
H, 4-H, 17-H), 1.40–1.12 (m, 8 H, 18-H to 21-H), 1.00–0.79 (m, 9
H, C-1, C-2, C-22) ppm. 13C NMR (150 MHz, CDCl3): δ = 173.7
[14] L. Murtagh, C. Dunne, G. Gabellone, N. J. Panesar, S. Field,
L. M. Reeder, J. Saenz, G. P. Smith, K. Kissick, C. Martinez,
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Wong, Org. Process Res. Dev. 2011, 15, 1315–1327.
(4 C, C-6, C-8, C-11, C-15), 69.2 (1 C, C-13), 65.8 (1 C, C-14), 52.4 [15] F. Sagui, C. De Micheli, G. Roda, P. Magrone, R. Pizzoli, S.
Riva, J. Mol. Catal. B 2012, 75, 27–34.
(2 C, C-5, C-7), 43.1, (1 C, C-12), 41.4 (1 C, C-4), 34.3 (1 C, C-
16), 31.7 (C-9, C-10, CH2), 29.0, 25.0, 22.7 (CH2), 22.9 (1 C, C-3),
22.0 (1 C, C-1, C-2), 14.2 (1 C, C-22) ppm. HRMS (ESI): calcd.
for C22H40N2O7Na 467.2733 [M + Na]+; found 467.2743.
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3999–4006.
21: Rf = 0.58 [CH2Cl2/MeOH (95:5, v/v) with Et3N (1%)]. [α]2D0
=
–9.9 (MeOH, 0.5 gdL–1). 1H NMR (600MHz, CDCl3): δ = 6.29 (t,
J12-NHC11 = 6.1 Hz, 1 H, NHC11), 6.25 (d, J5-NHC8 = 8.5 Hz, 1 H,
NHC8), 5.12–5.06 (m, 1 H, 13-H), 4.57 (td, J5-NHC8 = 8.5, J4-5
=
5.0 Hz, 1 H, 5-H), 4.26 (dd, J14a-14b = 12.1, J13-14a = 4 Hz, 1 H,
14a-H), 4.10 (dd, J14a-14b = 12.1, J13-14b = 5.9 Hz, 1 H, 14b-H),
3.72 (s, 3 H, 7-H), 3.54 (ddd, J12a-12b = 14.2, J12a-NHC11 = 6.1,
[21] L. Pellowska-Januszek, B. Dmochowska, E. Skorupa, J.
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2003, 24, 4810–4818.
J12a-13 = 5.1 Hz, 1 H, 12a-H), 3.40 (dt, J12a-12b = 14.2, J12b-13
=
J12b-NHC11 = 6.1 Hz, 1 H, 12b-H), 2.58–2.48 (m, 4 H, 9-H, 10-H),
2.32, 2.30 (2 t, J16-17 = 7.6 Hz, 4 H, 16-H), 1.73–1.47 (m, 7 H, 3-
H, 4-H, 17-H), 1.36–1.17 (m, 16 H, 18-H to 21-H), 0.93 (2 d, 6 H,
1-H, 2-H), 0.88 (2 t, J = 7.1, J = 7.3 Hz, 6 H, 22-H) ppm. 13C
NMR (150MHz, CDCl3): δ = 173.7, 173.6, 173.4, 172.4, 172.1 (5
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Pages: 11
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Received: April 14, 2014
Published Online: ᭿
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Date: 19-08-14 17:45:58
Pages: 11
Enzymatic Transesterification
Enzymatic Transesterification
C. Gervaise, V. Bonnet,* F. Nolay,
C. Cézard, I. Stasik, C. Sarazin,
F. Djedaïni-Pilard ............................ 1–11
Diesterification of 3-[(β-Cyclodextrinyl)-
succinamido]propane-1,2-diol Catalysed by
Lipase: Diastereoselectivity or Tridimen-
sional Substrate Specificity?
Two diastereoisomers of 3-[(β-cyclodex-
trinyl)succinamido]propane-1,2-diol have
been transesterified with fatty esters and
immobilized lipase from Mucor miehei.
They react with different conversions, con-
trary to their analogues without cyclodex-
trins. The hydrogen bonds in the three-di-
mensional structures, which account for
this phenomenon, are revealed by molecu-
lar modelling and NMR studies.
Keywords: Enzyme catalysis / Cyclodex-
trins / Lipids / Diastereoselectivity / Mo-
lecular modeling
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