Direct synthesis of maradolipids and other trehalose 6-monoesters and 6,6′-diesters

Article abstract of DOI:10.1021/jo302231v

It was shown that reaction of trehalose with 1 equiv of a fatty acid in pyridine promoted by 1 equiv of the uronium-based coupling agent 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU) at room temperature gives a good yield of the primary ester accompanied by small amounts of the diprimary ester using hexanoic, palmitic, and oleic acids as examples. Reactions using 2 equiv of the fatty acids gave the symmetrical diesters. The monoesters were reacted with different fatty acids to give nonsymmetric 6,6′-diesters in very good yields. Compounds synthesized include the most abundant component of the very complex maradolipid mixture, 6-O-(13-methyltetradecanoyl)-6′-O-oleoyltrehalose, and a component potentially present in this mixture, 6-O-(12-methyltetradecanoyl)-6′-O- oleoyltrehalose, a derivative of an ante fatty acid. The C5-C6 rotameric populations of 6-O-monoesters, symmetrical 6,6′-diesters, and 2,6,6′-triesters of fatty acids were calculated from the values of the H5-H6R and H5-H6S coupling constants and found to be similar to those found for glucose. The rotameric populations of the monosubstituted glucose residues in the 2,6,6′-triesters was altered considerably to favor the gt rotamer, presumably because of attraction between the 2- and 6′-fatty acid chains.

Full text of DOI:10.1021/jo302231v

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Article
Direct Synthesis of Maradolipids and Other
Trehalose 6-Monoesters and 6,6'-Diesters
Nawal K. Paul, Jean-d'Amour Karemerwa Twibanire, and T. Bruce Grindley
J. Org. Chem., Just Accepted Manuscript • DOI: 10.1021/jo302231v • Publication Date (Web): 10 Dec 2012
Downloaded from http://pubs.acs.org on December 15, 2012
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Direct Synthesis of Maradolipids and Other Trehalose 6ꢀMonoesters and 6,6'ꢀ
Diesters
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Nawal K. Paul, Jeanꢀd'Amour K. Twibanire, and T. Bruce Grindley*
Department of Chemistry, Dalhousie University, Halifax, NS, Canada B3H 4J3
bruce.grindley@dal.ca
Abstract
It was shown that reaction of trehalose with one equivalent of a fatty acid in pyridine promoted by
one equivalent of the uroniumꢀbased coupling agent 2ꢀ(1Hꢀbenzotriazoleꢀ1ꢀyl)ꢀ1,1,3,3ꢀ
tetramethyluronium tetrafluoroborate (TBTU) at room temperature gives a good yield of the primary
ester accompanied by small amounts of the diprimary ester, using hexanoic, palmitic, and oleic
acids as examples. Reactions using two equivalents of the fatty acids gave the symmetrical diesters.
The monoesters were reacted with different fatty acids to give nonꢀsymmetric 6,6'ꢀdiesters in very
good yields. Compounds synthesized include the most abundant component of the very complex
maradolipid mixture, 6ꢀOꢀ(13ꢀmethyltetradecanoyl)ꢀ6’ꢀOꢀoleoyltrehalose, and a component
potentially present in this mixture, 6ꢀOꢀ(12ꢀmethyltetradecanoyl)ꢀ6’ꢀOꢀoleoyltrehalose, a derivative
of an ante fatty acid. The C5ꢀC6 rotameric populations of 6ꢀOꢀmonoesters, symmetrical 6,6’ꢀ
diesters, and 2,6,6’ꢀtriesters of fatty acids were calculated from the values of the H5ꢀH6R and H5ꢀ
H6S coupling constants and found to be similar to those found for glucose. The rotameric
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populations of the monosubstituted glucose residues in the 2,6,6’ꢀtriesters was altered considerably
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to favor the gt rotamer, presumably because of attraction between the 2ꢀ and 6’ꢀfatty acid chains.
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Introduction
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Primary monoesters and diesters of trehalose (see Figure 1) have been of interest since the
recognition1 that they were important components of the outer membranes of mycobacteria, in
which the carboxylic acids are mycolic acids, complex longꢀchain βꢀhydroxy acids.2 They are also
of interest for many diverse biological activities.2b,3 Recently, they have been identified as
components of the outer membrane of dauer (enduring) larva of the wellꢀknown nematode,
caenorhabditis elegans.4 This form of larva appears when the nematode is exposed to extremely
dry conditions and the altered membrane allows the nematode to survive extreme desiccation.5 The
mixture of fatty acids present in the outer membrane, the "maradolipids",4 is extremely complex,
with about 38% of the fatty acids being monomethyl branched fatty acids and about 16% containing
cyclopropyl groups. The most abundant component is a nonsymmetric 6,6'ꢀtrehalose diester, 6ꢀOꢀ
(13ꢀmethylmyristoyl)ꢀ6’ꢀOꢀoleoyltrehalose.4 The only monomethyl branched fatty acids that have
been identified in C. elegans are branched next to the terminal carbon, that is, they are iso fatty
acids.4,6 Nevertheless, ante monomethyl branched fatty acids, that is, fatty acids branched on the
carbon second from the terminal carbon are common in nature.6b
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Figure 1. Trehalose 6,6'ꢀdiesters
There has been extensive effort directed at the synthesis of trehalose primary esters.2b,7 Most
authors have chosen to use protecting group strategies. One approach has been to use temporary
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protection of the primary hydroxyls with trityl or tertꢀbutyldimethylsilyl or tertꢀbutyldiphenylsilyl
groups before benzylation, removal of the primary protecting groups and acylation.8 The discovery9
that primary trimethylsilyl groups can be selectively removed by mild aqueous base has led to the
extensive use of the 2,2',3,3',4,4'ꢀhexaꢀOꢀtrimethylsilyl derivative for acylation studies.9ꢀ10 An
alternative strategy has been to selectively convert the primary hydroxyls into leaving groups, either
sulfonates11 or halides,9 before introducing acyl groups via SN2 substitution with carboxylate
salts.8a,8b,10a Trehalose has also been monoesterified at Oꢀ6 enzymically using a variety of vinyl
fatty acid esters in dimethyl formamide by a protease from Bacillus subtilis in good yields.12
Protectingꢀgroupꢀfree strategies are inherently attractive but few have been disclosed to this point.
Transesterification gave quite low yields.3a,13 Tributylstannylation gave moderate yields only when
the conditions using the toxic tributylstannyl ethers were carefully optimized.14 Mitsunobu
reactions are more attractive but again the yields are in the 50 to 60% range and the best solvent is
toxic hexamethylphosphoramide.15 This publication describes the protectingꢀgroupꢀfree synthesis
of 6ꢀmonoesters and 6,6'ꢀdiesters of trehalose using our primaryꢀselective acylation procedure,16
recently applied to the synthesis of a library of glycolipid antigens against Lyme disease.17
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Results and Discussion
Synthesis. The conditions developed for the regioselective acylation of primary alcohols in the
presence of secondary alcohols involved reaction of the diol or polyol with the carboxylic acid in
N,Nꢀdimethylformamide (DMF) with at least 2 equiv of diisopropylethylamine (DIEA) and 1.2
equiv of 2ꢀ(1Hꢀbenzotriazoleꢀ1ꢀyl)ꢀ1,1,3,3ꢀtetramethyluronium tetrafluoroborate (TBTU).16
Trehalose is relatively insoluble in DMF but it was found that pyridine was a good solvent for this
reaction as it was for the selective acylation of galactose.17 Reaction of trehalose (1) with a slight
excess of the fatty acid at room temperature gave good yields (65ꢀ69%) of the
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Scheme 1. Synthesis of trehalose 6ꢀmonoesters and 6,6'ꢀdiesters
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Table 1. Conditions and outcomes for the reactions of trehalose (1) with fatty acids
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entry
fatty acid
(equiv)
TBTU
(equiv)
DIEA
(equiv)
time (h)
temp
product, yield (%)
6ꢀmono
2a, 69
2a, 20
2a, 19
2a, 10
2b, 67
2b, 37
2b, 16
2b, 18
2c, 65
2c, 18
2c, 22
2c, 19
2c, 0
6,6’ꢀdi
3a, 14
3a, 63
3a, 63
3a, 48a
3b, 14
3b, 32
3b, 66
3b, 69
3c, 15
3c, 66
3c, 70
3c, 66
3c, 48a
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1
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hexanoic (1.1)
hexanoic (2.1)
hexanoic (2.1)
hexanoic (3.5)
palmitic (1.1)
palmitic (1.1)
palmitic (2.1)
palmitic (2.2)
oleic (1.1)
1.1
2.1
2.1
3.5
1.1
1.1
2.1
2.2
1.1
2.1
2.2
3.5
5.0
2.1
2.1
0
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rt
rt
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3.5
0
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0
48
40 oC
rt
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0
168
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rt
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rt
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oleic (2.1)
0
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rt
oleic (2.2)
0
192
60
rt
oleic (3.5)
0
rt
oleic (5.0)
0
168
rt
a Plus the 2,6,6'ꢀtriester (4) in the yield given in the experimental section.
6ꢀOꢀmonoacylated products as pictured in Scheme 1 and shown in entries 1, 5 and 9 of Table 1.
Under these conditions, small amounts of the 6,6'ꢀdiꢀOꢀacylated products are also obtained,
consistent with the first substitution having little effect on the reactivity of the second primary
hydroxyl group. The long chains of the fatty acids cause these reactions to be considerably slower
than the corresponding reactions with simple acids, such as benzoic acid, and longer reaction times
are required to achieve complete reaction of the fatty acids. As noted in the reactions with galactose
derivatives,17 the added base is unnecessary if the solvent is pyridine (compare entries 1, 5, and 9 in
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Table 1) consistent with the role of the base in the reactions of primary alcohols being to accept
protons released from the initial reaction of the acid with the uronium salt (TBTU) and in the
formation of the active ester on addition of the alcohol. Use of two or more equivalents of fatty
acids gives reasonable yields of the 6,6’ꢀdiꢀOꢀacyl products (see entries 2, 3, 7, and 10 in Table 1).
Neither increasing the relative amount of fatty acid beyond 2.1 equiv nor raising the reaction
temperature improved the yields of the disubstituted products. Instead additional products were
obtained of which the 2,6,6’ꢀtriester (4) (Figure 2) was the most prominent, isolated in 20% yield
from the reaction of trehalose with 3.5 equiv of hexanoic acid for 48 h and in 40% yield from the
reaction with 5 equiv of oleic acid for 168 h. No products of esterification on secondary oxygen
atoms have previously been observed in reactions of this type with monosaccharides. Perhaps the
hydroxyl group at Oꢀ2 of trehalose is more acidic than hydroxyls of monosaccharides because the
anomeric oxygen is more electron withdrawing in a nonꢀreducing disaccharide.
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Figure 2. Structure of triester products
The most abundant component in the maradolipid mixture is 6ꢀOꢀ(13ꢀmethylmyristoyl)ꢀ6’ꢀ
Oꢀoleoyltrehalose (5b).4 It was found that unsymmetrical derivatives of this type could be
synthesized in good yields by reacting the monooleoyl derivative 2c with 1.1 equiv of the fatty acid
for extended reaction times at room temperature (see Scheme 2 and entries 4 and 6 of Table 2).
Branched fatty acids such as 13ꢀmethylmyristic acid are available commercially from specialized
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companies at great expense for the amounts necessary for synthetic purposes but here this acid was
synthesized by the method of Foglia and Vail.18 Compound 5b has been synthesized previously in
fiveꢀstep routes using TMS ethers as temporary protecting groups.10h,10i
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Scheme 2. Synthesis of nonsymmetric trehalose 6,6'ꢀdiesters
Table 2. Conditions and outcomes for the reactions of 6ꢀOꢀoleoyltrehalose (2c) with fatty
acids
entry
fatty acid
(equiv)
hexanoic (1.1)
TBTU time (h)
(equiv)
isolated
yield (%)
59a
1
2
3
4
5
6
1.1
1.1
1.1
1.1
1.1
1.1
72
13ꢀmethyltetradecanoic (1.1)
13ꢀmethyltetradecanoic (1.1)
13ꢀmethyltetradecanoic (1.1)
12ꢀmethyltetradecanoic (1.1)
12ꢀmethyltetradecanoic (1.1)
72
62b
72c
81
120
170
72
54d
79
170
b
a26% 2c also isolated. 22% 2c also isolated. c10% 2c also isolated. d27% 2c also isolated.
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The ante derivative 6ꢀOꢀ(12ꢀmethyltetradecanoyl)ꢀ6’ꢀOꢀoleoyltrehalose (5c) was prepared in
the same way (see Scheme 2) from 2c and 12ꢀmethylmyristic acid. This fatty acid was prepared in
racemic form using a Wittig reaction of the Wittig reagent derived from 11ꢀbromoundecanoic acid
with 2ꢀbutanone followed by hydrogenation as previously.19 Compound 5c had never been
synthesized previously and provides a sample for examining whether such compounds are part of
the complex maradolipid mixture.
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Conformational analysis. It was also of interest to determine whether the diverse biological
activities of these compounds are influenced by alteration of the populations of the rotameric
conformations adopted by the hydroxymethyl groups of trehalose caused by the interactions of the
hydrophobic fatty acid alkyl groups. Trehalose itself adopts a conformation with both anomeric
linkages adopting normal exoꢀanomeric conformations both in the solidꢀstate20 and in solution.21
Hydroxymethyl rotameric populations have been discussed extensively22 and have been determined
carefully for glucose derivatives by making use of all H,H and C,H coupling constants of
isotopically enriched derivatives (see Figure 3 for definition of rotamer names).23 4,6ꢀUnsubstituted
derivatives slightly prefer the gt conformer over the gg conformer with the tg conformer having a
population of about 10% or slightly less.22d,23 Barnett and Naidoo suggested that the preference for
the gt conformer is due to direct and waterꢀmediated hydrogen bonds between the O6 hydroxyl
hydrogen and O5.22c In the solid state,20 trehalose and its dihydrate are present in conformations
where the two hydroxymethyl groups each adopt one of the two rotamers populated in solution, the
13
gg and gt rotamers, giving rise to C CP/MAS spectra with one signal for each of the 12 carbon
atoms.20a,24
H5ꢀH6 vicinal coupling constants were determined for the three monoesters (2), the three
symmetrical diesters (3), and the two 2,6,6’ꢀtriesters (4) making the reasonable (all ꢁν/J >6)
assumption that the coupling patterns were first order. The values obtained are reported in Table 3.
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The hydroxymethyl groups can adopt three conformers, termed the gg, gt, and tg rotamers,
according to whether O5 and C4, respectively, are gauche or trans to O6 (see Figure 3). The
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Figure 3. Newman projections from C5 to C6 illustrating the definitions of the three
hydroxymethyl rotamers and atom labeling.
coupling constants were used to calculate rotameric populations using the values of the coupling
constants for each rotamer calculated by Stennutz et al23a (see supporting information for the
details). The percentage populations for each rotamer are listed in Table 4. The percentage
populations for the monoesters and diesters are similar to those obtained for glucose
previously22d,23a although the relative amounts of the gg rotamer appears to have increased slightly
at the expense of the gt conformer. This is consistent with loss of the stabilizing effect for the gt
rotamer of direct and hydroxylic solvent mediated hydrogen bonds between the O6 hydroxyl
hydrogen and O5.22c Therefore, aggregation of the long lipophilic groups on O6 does not appear to
influence the rotameric populations for the hydroxymethyl groups of these two classes of
compounds significantly.
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Table 3. Threeꢀbond coupling constants observed for the C6 protons
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6
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Compound
3J5,6R (Hz)
4.98
3J5,6S (Hz)
2.01
3J5',6'R (Hz)
5.31
3J5',6'S (Hz)
1.87
2a
2b
2c
3a
3b
3c
4a
4c
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5.08
2.00
5.53
2.04
5.05
2.06
5.48
2.16
5.20
2.11
5.20
2.11
5.29
2.05
5.29
2.05
5.28
2.11
5.28
2.11
4.96
2.09
7.43
2.07
4.87
2.07
7.90
1.89
Table 4. Percentage populations of rotamers
compound
% gt for
% gg for
% tg for
% gt for
% gg for
% tg for
C5C6 bond
C5C6 bond
C5C6 bond C5'C6' bond C5'C6' bond C5'C6' bond
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6.6
6.4
7.1
7.6
6.9
48
41
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44
47
47
5.0
6.9
8.4
7.6
6.9
2a
2b
2c
3a
3b
46
43
42
46
50
51
7.6
7.4
7.4
46
70
76
46
23
19
7.6
6.6
4.6
3c
4a
4c
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In contrast, for the 2,6,6’ꢀtriesters, the two sets of H5ꢀH6 coupling constants were different;
the set for the disubstituted glucose unit was similar to those observed for the mono and diesters but
for the monosubstituted glucose residue, the J5',6'R value was between 2.2 and 2.7 Hz larger than
those observed for all other residues. For this residue, the gt conformer was calculated to be more
favored, to the extent of 70 and 76% of the rotamers for 4a and 4c, respectively, mostly at the
expense of the gg conformer. This change probably is caused by intramolecular van der Waals
interactions between the long chains of the 6’ꢀester and the 2ꢀester, favoring the rotamer where the
C6’ ester is turned toward the disubstituted glucose residue, as illustrated in Figure 4.
O
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O
O
HO
O
HO
HO
O
O
O
OH
R
O
O
OH
Figure 4. A conformation of 4a (R = C5H11) illustrating how adopting the gt conformation for C5ꢀ
C6 bond in the monosubstituted glucose ring allows van der Waals interactions between the long
chains of the 6’ꢀester and the 2ꢀester
Conclusions
In summary, TBTUꢀpromoted esterification of trehalose with one equivalent of fatty acids
provides 6ꢀOꢀmonoesters in one step in good yields (~70%); two equivalents provides symmetrical
6,6'ꢀdiesters in fair yields. In comparison, enzymic esterification using a commercially available
protease from Bacillus subtilis gave the monopalmitate 2b in 84% yield but the monooleaoate 2c in
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55% yield using a quite long reaction time (12 days).12 Other protectingꢀgroupꢀfree chemical
methods give monoesters in lower yields, of which, the Mitsunobu reaction is most efficient (47ꢀ
61% yields).15 The monoesters can be again monoesterified in the same way to provide nonꢀ
symmetric 6,6'ꢀOꢀdiesters in two steps from trehalose in very good yields. Some of the diesters
present in the complex mixture of maradolipids have been synthesized and the method allows
expeditious synthesis of any desired structure.
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Experimental Methods
General esterification procedures using TBTU
A. Use of trehalose (1). In an ovenꢀdried roundꢀbottomed flask equipped with a magnetic stir bar, a
fatty acid (number of equiv given in Table 1) and TBTU (number of equiv given in Table 1) were
dissolved in anhydrous pyridine (5 mL) and the resulting mixture was stirred at rt for 30 min under a
nitrogen atmosphere. A solution of trehalose (amount used listed with the individual products) in
dry pyridine (3 mL) was then injected into the reaction mixture via syringe and stirring was
continued at the temperature and for the time given in Table 1. Pyridine was removed under vacuum
and the resulting residue was purified using silica gel column chromatography with elution using a
solvent gradient of 5 ꢀ 25% methanol in EtOAc ꢀ DCM (1:1).
B. Use of 6ꢀOꢀoleoylꢀα,αꢀtrehalose (2c). In an ovenꢀdried roundꢀbottomed flask equipped with a
magnetic stir bar, a fatty acid (1.1 equiv) and TBTU (1.1 equiv) were dissolved in anhydrous
pyridine (5 mL) and the resulting mixture was stirred at rt for 30 min under a nitrogen atmosphere.
A solution of 6ꢀOꢀoleoylꢀα,αꢀtrehalose (2c) (amount used given with individual products) in dry
pyridine (3 mL) was then injected into the reaction mixture via syringe and stirring was continued at
rt for the time given in Table 2. Pyridine was removed under vacuum and the resulting residue was
dissolved in EtOAcꢀTHF (3:1, 20 mL). This solution was washed with saturated NaHCO3 (2 x 3
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mL), dried (MgSO4), and concentrated to give a crude product which was purified using silica gel
column chromatography with elution using a gradient of 5 ꢀ 25% methanol in EtOAc ꢀ DCM (1:1).
6ꢀOꢀHexanoylꢀα,αꢀtrehalose (2a). The title compound was synthesized using procedure A above
with trehalose (1) (200 mg, 0.58 mmol) and hexanoic acid under conditions listed in Table 1, entry 1
and was obtained as a colorless solid (178 mg, 69% yield: RF 0.20 [25% MeOH in EtOAcꢀ
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o
o
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DCM(1:1), v/v], mp 136 – 138 C, lit8g mp 135ꢀ137 C; H NMR (CD3OD) δ 0.91 (t, 3H, J = 7.0
Hz, Me), 1.28 ꢀ 1.36 (m, 4H, 2 x CH2), 1.61 (m, 2H, CH2CH3), 2.33 (t, 2H, J = 7.5 Hz, CH2CO),
3.34 ꢀ 3.38 (m, 2H, Hꢀ4, Hꢀ4'), 3.49 (dd, 2H, J = 3.9 Hz, 7.9 Hz, Hꢀ2, Hꢀ2'), 3.67 (dd, 1H, J = 5.2
Hz, 11.7 Hz, Hꢀ6'R), 3.79 ꢀ 3.83 (m, 4H, Hꢀ3, Hꢀ3', Hꢀ6', Hꢀ5'), 4.02 (ddd, 1H, J4,5 = 10.1 Hz, J5,6R
=
5.2 Hz, J5,6S = 2.1 Hz, Hꢀ5), 4.20 (dd, 1H, J = 5.2 Hz, 11.9 Hz, Hꢀ6R), 4.38 (dd, 1H, J = 2 Hz, 11.9
Hz, Hꢀ6S), 5.08 (d, 1H, J = 3.8 Hz, Hꢀ1'), 5.10 (d, 1H, J = 3.7 Hz, Hꢀ1); 13C NMR δ 175.6 (C=O),
95.3, 95.2 (Cꢀ1, Cꢀ1'), 74.8, 74.6, 74.0, 73.33, (Cꢀ2, Cꢀ2', Cꢀ3, Cꢀ3'), 73.3, 72.0, 71.6 (Cꢀ4, Cꢀ4', Cꢀ
5, Cꢀ5'), 64.5, 62.7 (Cꢀ6, Cꢀ6'), 35.1, 32.6, (COCH2, COCH2CH2), 32.5, 26.0 (hexanoyl CH2), 23.6
(CH2CH3), 14.4 (Me); HR ESI MS m/z calc for C18H32NaO12 : 463.1786; found: 463.1764.
In addition, some of compound 3a (44 mg, 14% yield) was obtained.
6,6'ꢀDiꢀOꢀhexanoylꢀα,αꢀtrehalose (3a). The title compound was synthesized using procedure A
above with trehalose (200 mg, 0.58 mmol) and hexanoic acid under conditions listed in Table 1,
entry 2 and was obtained as a colorless solid (198 mg, 63% yield): RF 0.40 [20% MeOH in EtOAcꢀ
DCM(1:1), v/v], mp = 157 – 160 oC, lit8g mp 157.7ꢀ159.0 oC; 1H NMR (CD3OD) δ 0.91 (t, 6H, J =
6.0 Hz, 2 x Me), 1.30 ꢀ 1.37 (m, 8H, 4 x CH2), 1.62 (m, 4H, CH2CH3), 2.34 (t, 4H, J = 7.0 Hz,
CH2CO), 3.33 (dd, 2H, J = 9.0 Hz, 10.0 Hz, Hꢀ4, Hꢀ4'), 3.47 (dd, 2H, J = 3.7 Hz, 9.7 Hz, Hꢀ2, Hꢀ
2'), 3.77 (dd, 2H, J = 9.1 Hz, 9.6 Hz, Hꢀ3, Hꢀ3'), 4.01 (ddd, 2H, J4,5 = 10.0 Hz, J5,6R = 5.2 Hz, J5,6S
=
2.1 Hz, Hꢀ5, Hꢀ5'), 4.19 (dd, 2H, J = 5.2 Hz, 11.9 Hz, Hꢀ6R), 4.35 (dd, 2H, J = 2.1 Hz, 11.9 Hz, Hꢀ
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6S), 5.03 (d, 2H, J = 3.7 Hz, Hꢀ1',Hꢀ1); 13C NMR δ 175.5 (C=O), 95.3 (Cꢀ1, Cꢀ1'), 74.5, 73.1 (Cꢀ2,
Cꢀ2', Cꢀ3, Cꢀ3'), 71.9, 71.5 (Cꢀ4, Cꢀ4', Cꢀ5, Cꢀ5'), 64.4 (Cꢀ6, Cꢀ6'), 35.0, 32.4 (COCH2,
COCH2CH2), 25.8 (hexanoyl CH2), 23.4 (CH2CH3), 14.3 (Me); HR ESI MS m/z calc for
C24H42NaO13: 561.2518; found: 561.2517.
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In addition, some of compound 2a (49 mg, 20% yield) was obtained.
2,6,6'ꢀTriꢀOꢀhexanoylꢀα,αꢀtrehalose (4a). Following procedure A above using trehalose (325 mg,
0.95 mmol) with 3.5 equiv of hexanoic acid (385.8 mg) as in Table 1 entry 4, the reaction gave
compounds 2a ( mg, 19% yield) and 3a ( mg, 48% yield) plus the title compound as a colorless
1
syrup (119 mg, 20% yield): RF 0.60 [5% MeOH in EtOAcꢀDCM(1:1), v/v]; H NMR (CD3OD) δ
0.91 (t, 9H, J = 6.0 Hz, 3 x Me), 1.30 ꢀ 1.39 (m, 12H, 6 x CH2), 1.59ꢀ1.63 (m, 6H, CH2CH3), 1.95ꢀ
2.45 (m, 6H, CH2CO), 3.27 (dd, 1H, J = 9.0 Hz, 10.0 Hz, Hꢀ4'), 3.43 (dd, 1H, J = 9.1 Hz, 10.0 Hz,
Hꢀ4), 3.47 (dd, 1H, J = 3.8 Hz, 9.8 Hz, Hꢀ2'), 3.69 (t, 1H, J = 9.0 Hz , Hꢀ3'), 3.77 (ddd, 1H, J4',5' =
9.6 Hz, J5',6'R = 7.2 Hz, J5',6'S = 2.0 Hz, Hꢀ5'), 3.99 (t, 1H, J = 9.1 Hz, Hꢀ3), 4.04 (ddd, 1H, J4,5 = 7.0
Hz, J5,6R = 4.9 Hz, J5,6S = 2.0 Hz, Hꢀ5), 4.16 (dd, 1H, J = 7.0 Hz, 11.9 Hz, Hꢀ6'R), 4.23 (dd, 1H, J =
5.0 Hz, 12.0 Hz, Hꢀ6R), 4.29 (dd, 1H, J = 2.0 Hz, 11.8 Hz, Hꢀ6'S), 4.39 (dd, 1H, J = 2.0 Hz, 12.0
Hz, Hꢀ6S), 4.70 (dd, 1H, J = 3.6 Hz, 10.0 Hz, Hꢀ2), 5.02 (d, 1H, J = 3.7 Hz, Hꢀ1'), 5.18 (d, 1H, J =
13
3.6 Hz, Hꢀ1); C NMR δ 175.52, 175.44, 174.8 (C=O), 95.3, 92.6 (Cꢀ1, Cꢀ1'), 74.8, 74.2, 73.0,
72.94 (Cꢀ2, Cꢀ2', Cꢀ3, Cꢀ3'),72.09, 72.04, 71.9, 71.6 (Cꢀ4, Cꢀ4', Cꢀ5, Cꢀ5'), 64.9, 64.2 (Cꢀ6, Cꢀ6'),
39.0, 35.1, 35.0, 34.96, 32.55, 32.52 (COCH2, COCH2CH2), 25.9, 25.8, 25.76, 25.73, 25.66
(hexanoyl CH2), 23.5 (CH2CH3), 14.5 (Me); HR ESI MS m/z calc for C30H52NaO14: 659.3249;
found: 659.3240.
6ꢀOꢀPalmitoylꢀα,αꢀtrehalose (2b). The title compound was synthesized using procedure A above
with trehalose (300 mg, 0.87 mmol) and palmitic acid under conditions listed in Table 1, entry 5 and
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was obtained as a colorless solid (341 mg, 67% yield): RF 0.33 [20% MeOH in EtOAcꢀDCM(1:1)],
5
6
7
o
o
1
mp 156 – 159 C, lit12 mp 114ꢀ/116 C
.
; H NMR (CD3OD) δ 0.90 (t, 3H, J = 6.0 Hz, Me), 1.29 ꢀ
8
9
1.37 (m, 24H, 12 x CH2), 1.62 (m, 2H, CH2CH3), 2.34 (t, 2H, J = 7.2 Hz, CH2CO), 3.30 ꢀ 3.33 (m,
2H, Hꢀ4, Hꢀ4'), 3.46, 3.47 (2 overlapping dd, 2H, J1,2 = 4.0 Hz, J2,3 = 9 Hz, Hꢀ2, Hꢀ2'), 3.67 (dd, 1H,
J = 5.7 Hz, 12.1 Hz, Hꢀ6'R), 3.76 ꢀ 3.83 (m, 4H, Hꢀ3, Hꢀ3', Hꢀ6', Hꢀ5'), 4.01 (ddd, 1H, J4,5 = 10.1
Hz, J5,6R = 5.1 Hz, J5,6S = 2.0 Hz, Hꢀ5), 4.19 (dd, 1H, J = 5.1 Hz, 11.9 Hz, Hꢀ6R), 4.35 (dd, 1H, J =
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2.0 Hz, 11.9 Hz, Hꢀ6S), 5.07 (d, 1H, J = 3.7 Hz, Hꢀ1'), 5.10 (d, 1H, J = 3.7 Hz, Hꢀ1); C NMR δ
175.6 (C=O), 95.4, 95.3 (Cꢀ1, Cꢀ1'), 74.8, 74.6, 74.1, 73.2, (Cꢀ2, Cꢀ2', Cꢀ3, Cꢀ3'), 73.4, 72.1, 71.6
(Cꢀ4, Cꢀ4', Cꢀ5, Cꢀ5'), 64.5, 62.8 (Cꢀ6, Cꢀ6'), 35.2, (COCH2, COCH2CH2), 33.2, 30.9, 30.8, 30.6,
30.6, 30.4, 26.0 (palmitoyl CH2), 23.9 (CH2CH3), 14.6 (Me); HR ESI MS m/z calc for
C28H52NaO12: 603.3351; found: 603.3335.
In addition, some of compound 3b (71 mg, 14% yield) was obtained.
6,6'ꢀDiꢀOꢀpalmitoylꢀα,αꢀtrehalose (3b). The title compound was synthesized using procedure A
above with trehalose (300 mg, 0.87 mmol) and palmitic acid under conditions listed in Table 1,
entry 7 and was obtained as a gummy solid (480 mg, 66% yield): RF 0.62 [20% MeOH in EtOAcꢀ
1
DCM(1:1), v/v] ; H NMR (CD3OD) δ 0.90 (t, 6H, J = 6.0 Hz, 2 x Me), 1.26 ꢀ 1.39 (m, 48H, 24 x
CH2), 1.61 (m, 4H, CH2CH3), 2.34 (t, 4H, J = 7.2 Hz, CH2CO), 3.30 ꢀ 3.34 (m, 2H, Hꢀ4, Hꢀ4'), 3.47
(dd, 2H, J = 3.7 Hz, 9.7 Hz, Hꢀ2, Hꢀ2'), 3.77 (t, 2H, J = 9.3 Hz, Hꢀ3, Hꢀ3'), 4.02 (ddd, 2H, J4,5
=
10.0 Hz, J5,6R = 5.3 Hz, J5,6S = 2.1 Hz, Hꢀ5, Hꢀ5 ), 4.19 (dd, 2H, J = 5.3 Hz, 11.9 Hz, Hꢀ6R, Hꢀ6'R),
'
13
4.35 (dd, 2H, J = 2.1 Hz, 11.9 Hz, Hꢀ6S, Hꢀ6'S), 5.04 (d, 2H, J = 3.7 Hz, Hꢀ1,Hꢀ1'); C NMR δ
175.5 (C=O), 95.4 ( Cꢀ1, Cꢀ1'), 74.7, 73.3 (Cꢀ2, Cꢀ2', Cꢀ3, Cꢀ3'), 72.1, 71.7 (Cꢀ4, Cꢀ4', Cꢀ5, Cꢀ5'),
64.6 (Cꢀ6, Cꢀ6'), 35.2, 33.2 (COCH2, COCH2CH2), 30.96, 30.93, 30.8, 30.6, 30.6, 30.4, 26.2
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(palmitoyl CH2), 23.9 (CH2CH3), 14.6 (Me); HR ESI MS m/z calc for C44H82NaO13: 841.5648;
found: 841.5648.
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In addition, some of compound 2b (82 mg, 16% yield) was obtained.
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6ꢀOꢀOleoylꢀα,αꢀtrehalose (2c). The title compound was synthesized using procedure A above with
trehalose (300 mg, 0.87 mmol) and oleic acid under conditions listed in Table 1, entry 9 and was
obtained as a colorless solid (346 mg, 65% yield): RF 0.37 [20 % MeOH in EtOAcꢀDCM(1:1), v/v],
o
o
o
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mp become transparent at 120 – 130 C, melted at 166 – 168 C (lit12 mp 165ꢀ167 C) ; H NMR
(CD3OD) δ 0.90 (t, 3H, J = 6.5 Hz, Me), 1.25 ꢀ 1.40 (m, 20H, 10 x CH2), 1.45 ꢀ 1.61 (m, 2H,
CH2CH3), 2.02 ꢀ 2.04 (m, 4H, 2 x CH2CHCH), 2.34 (t, 2H, J = 7.0 Hz, CH2CO), 3.30 ꢀ 3.32 (m, 2H,
Hꢀ4, Hꢀ4'), 3.46, 3.47 (2 overlapping dd, 2H, J1,2 = 3.9 Hz, J2,3 = 8.8 Hz, J1',2' = 4.0 Hz, J2',3' = 9.2
Hz, Hꢀ2, Hꢀ2'), 3.67 (dd, 1H, J = 5.5 Hz, 12.1 Hz, Hꢀ6'R), 3.76 ꢀ 3.85 (m, 4H, Hꢀ3, Hꢀ3', Hꢀ6S', Hꢀ
5'), 4.02 (ddd, 1H, J4,5 = 10.1 Hz, J5,6 = 5.1 Hz, J5,6 = 2.1 Hz, Hꢀ5), 4.20 (dd, 1H, J = 5.1 Hz, 11.9
Hz, Hꢀ6R), 4.36 (dd, 1H, J = 2.1 Hz, 11.9 Hz, Hꢀ6S), 5.07 (d, 1H, J = 3.7 Hz, Hꢀ1'), 5.09 (d, 1H, J =
13
3.7 Hz, Hꢀ1), 5.35 (t, 2H, J = 4.8 Hz, CH=CH); C NMR δ 175.4 (C=O), 130.9, 130.8 (CH=CH),
95.2, 95.1 (Cꢀ1, Cꢀ1'), 74.6, 74.4, 73.9, 73.19, 73.16, 71.91, 71.86, 71.4 (Cꢀ2, Cꢀ2', Cꢀ3, Cꢀ3', Cꢀ4,
Cꢀ4', Cꢀ5, Cꢀ5'), 64.4, 62.6 (Cꢀ6, Cꢀ6'), 35.0, 33.1 (COCH2, COCH2CH2), 30.8, 30.6, 30.5, 30.3,
30.2, 28.1, 26.0 (oleoyl CH2), 23.7 (CH2CH3), 14.5 (Me); HR ESI MS m/z calc for C30H54NaO12
629.3507; found: 629.3527.
In addition, some of compound 3c (115 mg, 15% yield) was obtained.
6,6'ꢀDiꢀOꢀoleoylꢀα,αꢀtrehalose (3c). The title compound was synthesized using procedure A above
with trehalose (150 mg, 0.43 mmol) and oleic acid under conditions listed in Table 1, entry 10 and
was obtained as a gummy solid (260 mg, 66% yield): RF 0.63 [20% MeOH in EtOAcꢀDCM(1:1),
v/v]; 1H NMR (CD3OD) δ 0.94 (t, 6H, J = 6.5 Hz, 2 x Me), 1.32 ꢀ 1.45 (m, 40H, 20 x CH2), 1.64 ꢀ
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1.67 (m, 4H, CH2CH3), 2.05 ꢀ 2.07 (m, 8H, CH2CHCH), 2.38 (t, 4H, J = 7.0 Hz, CH2CO), 3.33 (dd,
2H, J3,4 = 8.9 Hz, J4,5 = 10.1 Hz, Hꢀ4, Hꢀ4'), 3.50 (dd, 2H, J1,2 = 3.8 Hz, J2,3 = 9.7 Hz, Hꢀ2, Hꢀ2'),
3.81 (dd, 2H, J3,4 = 9.1 Hz, J2,3 = 9.5 Hz, 2H, Hꢀ3, Hꢀ3'), 4.05 (ddd, 2H, J4,5 = 10.1 Hz, J5,6 = 5.1
Hz, J5,6' = 2.1 Hz, Hꢀ5, Hꢀ5'), 4.23 (dd, 2H, J = 5.1 Hz, 11.9 Hz, Hꢀ6R, Hꢀ6’R), 4.39 (dd, 2H, J = 2.1
Hz, 11.9 Hz, Hꢀ6S, Hꢀ6’S), 5.09 (d, 2H, J = 3.8 Hz, Hꢀ1',Hꢀ1), 5.35 (t, 4H, J = 4.8 Hz, 4H,
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CH=CH); C NMR δ 175.5 (C=O), 131.1, 131.0 (CH=CH), 95.3, ( Cꢀ1, Cꢀ1'), 74.7, 73.3, 72.1,
71.6 (Cꢀ2, Cꢀ2', Cꢀ3, Cꢀ3', Cꢀ4, Cꢀ4', Cꢀ5, Cꢀ5'), 64.6 (Cꢀ6, Cꢀ6'), 35.2, 33.2 (COCH2, COCH2CH2),
31.0, 31.0, 30.8, 30.6, 30.5, 30.4, 28.3, 26.2 (oleoyl CH2), 23.9 (CH2CH3), 14.6 (Me); HR ESI MS
m/z calc for C48H86NaO13: 893.5961; found: 893.5950.
In addition, some compound 2c (48 mg, 18% yield) was obtained.
2,6,6'ꢀTriꢀOꢀoleoylꢀα,αꢀtrehalose (4c). The title compound was synthesized from trehalose (150
mg, 0.43 mmol) and 5 equiv of oleic acid (610 mg, 2.16 mmol) using procedure A under the
conditions of Table 1 entry 13. Compound 3c ( mg, 48% yield) was obtained plus the title
compound as a thick colorless syrup (198 mg, 40% yield): RF 0.43 [1% MeOH in EtOAcꢀ
1
DCM(1:1), v/v]; H NMR (CD3OD) δ 0.90 (t, 9H, J = 6.5 Hz, 3 x Me), 1.19 ꢀ 1.35 (m, 60H, 30 x
CH2), 1.57ꢀ1.66 (m, 6H, CH2CH3), 1.99 ꢀ 2.07 (m, 12H, CH2CHCH), 2.32ꢀ2.46 (m, 6H, CH2CO),
3.24 (dd, 1H, J = 9.0 Hz, 10.0 Hz, Hꢀ4'), 3.44 (overlapped dd, 1H, Hꢀ4), 3.47 (dd, 1H, J = 3.8 Hz,
10.0 Hz, Hꢀ2'), 3.68 (t, 1H, J = 9.2 Hz, Hꢀ3'), 3.78 (ddd, 1H, J4',5' = 10.0 Hz, J5',6'R = 7.7 Hz, J5',6'S
=
2.0 Hz, Hꢀ5'), 3.99 (t, 1H, J = 9.6 Hz, Hꢀ3), 4.07 (ddd, 1H, J4,5 = 7.0 Hz, J5,6R = 4.5 Hz, J5,6S = 2.2
Hz, Hꢀ5), 4.15 (dd, 1H, J = 7.8 Hz, 11.7 Hz, Hꢀ6'R), 4.24 (dd, 1H, J = 5.0 Hz, 12.0 Hz, Hꢀ6R), 4.29
(dd, 1H, J = 1.9 Hz, 12.0 Hz, Hꢀ6'S), 4.39 (dd, 1H, J = 2.2 Hz, 12.0 Hz, Hꢀ6S), 4.70 (dd, 1H, J = 3.7
Hz, 10.0 Hz, Hꢀ2), 5.01 (d, 1H, J = 3.7 Hz, Hꢀ1'), 5.18 (d, 1H, J = 3.6 Hz, Hꢀ1), 5.33ꢀ5.36 (m, 6H,
13
CH=CH); C NMR δ 175.52, 175.38, 174.8 (C=O), 131.06, 131.02 (CH=CH), 95.3, 92.7 (Cꢀ1, Cꢀ
1'), 74.9, 74.2, 73.0, 72.28 (Cꢀ2, Cꢀ2', Cꢀ3, Cꢀ3'),72.06, 72.00, 71.9, 71.6 (Cꢀ4, Cꢀ4', Cꢀ5, Cꢀ5'),
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65.0, 64.2 (Cꢀ6, Cꢀ6'), 35.3, 35.1, 35.0 (COCH2, COCH2CH2), 33.2, 31.0, 30.8, 30.7, 30.6, 30.55,
30.47, 30.40. 28.3, 26.3, 26.2, 26.0 (oleoyl CH2), 24.0 (CH2CH3), 14.7 (Me); HR ESI MS m/z calc
for C66H118NaO14 1157.8414; found 1157.8451.
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6ꢀOꢀHexanoylꢀ6'ꢀOꢀoleoylꢀα,αꢀtrehalose (5a). The title compound was synthesized using
procedure B above with 6ꢀOꢀoleoylꢀα,αꢀtrehalose (2c) (100 mg, 0.17 mmol) and hexanoic acid (21
mg, 0.18 mmol) and was obtained as a gummy solid (69 mg, 59% yield): RF 0.46 [20% MeOH in
EtOAcꢀDCM(1:1), v/v]; 1H NMR (CD3OD) δ 0.89 – 0.93 (m, 6H, 2 x Me), 1.25 ꢀ 1.41 (m, 24H, 12
x CH2), 1.61 ꢀ 1.64 (m, 4H, CH2CH3), 2.01 ꢀ 2.04 (m, 4H, CH2CHCH), 2.34 (t, 4H, J = 7.3 Hz,
CH2CO), 3.32 ꢀ 3.35 (m, 2H, Hꢀ4, Hꢀ4'), 3.46 (dd, 2H, J = 3.8 Hz, 9.7 Hz, Hꢀ2, H2'), 3.77 (t, 2H, J
= 9.3 Hz, Hꢀ3, Hꢀ3'), 3.99 ꢀ 4.03 (m, 2H, Hꢀ5, Hꢀ5'), 4.21 (dd, 2H, J = 5.4 Hz, 11.8 Hz, Hꢀ6R, Hꢀ
6’R), 4.35 (dd, 2H, J = 2.0 Hz, 11.9 Hz, Hꢀ6S, Hꢀ6'S), 5.04 (d, 2H, J = 3.8 Hz, Hꢀ1',Hꢀ1), 5.35 (t,
2H, J = 4.8 Hz, CH=CH); 13C NMR δ 175.62, 175.61 (C=O), 131.12, 130.97 (CH=CH), 95.4 (Cꢀ1,
Cꢀ1'), 74.7, 73.3 (Cꢀ2, Cꢀ2', Cꢀ3, Cꢀ3'), 72.1, 71.6 (Cꢀ4, Cꢀ4', Cꢀ5, Cꢀ5'), 64.6 (Cꢀ6, Cꢀ6'), 35.2, 33.2
(COCH2, COCH2CH2), 32.6, 30.99, 30.95, 30.8, 30.6, 30.5, 30.4, 30.3, 28.3, 26.2, 25.9 (CH2), 23.9,
23.5 (CH2CH3), 14.6, 14.4 (Me); HR ESI MS m/z calc for C36H64NaO13: 727.4239; found:
727.4216.
6ꢀOꢀ(13ꢀMethyltetradecanoyl)ꢀ6'ꢀOꢀoleoylꢀα,α–trehalose (5b). The title compound was
synthesized using procedure B above and the conditions in Table 2, entry 4 with 6ꢀOꢀoleoylꢀα,α–
trehalose (150 mg, 0.24 mmol) and 13ꢀmethyltetradecanoic acid (66 mg, 0.27 mmol), prepared
using a literature method.18 A gummy solid (168 mg, 81% yield): RF 0.51 [20% MeOH in EtOAcꢀ
DCM(1:1), v/v]; 1H NMR (CD3OD) δ 0.88 ꢀ 092 (m, 9H, 3 x Me), 1.06 ꢀ 1.12 (m, 2H, CH2), 1.25 ꢀ
1.35 (m, 36H, 18 x CH2), 1.53 (sept, 1H, J = 6.6 Hz, CH(CH3)2), 1.60 ꢀ 1.64 (m, 4H, CH2CH3),
2.01 ꢀ 2.05 (m, 4H, CH2CHCH), 2.34 (t, 4H, J = 7.4 Hz, CH2CO), 3.31 ꢀ 3.35 (m, 2H, Hꢀ4, Hꢀ4'),
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3.43 (dd, 2H, J = 3.7 Hz, 9.8 Hz, Hꢀ2, Hꢀ2'), 3.78 (t, 2H, J = 9.5 Hz, Hꢀ3, Hꢀ3'), 3.99 ꢀ 4.03 (m, 2H,
Hꢀ5, Hꢀ5'), 4.20 (dd, 2H, J = 5.3 Hz, 11.8 Hz, Hꢀ6R, Hꢀ6'R), 4.35 (dd, 2H, J = 1.7 Hz, 11.8 Hz, Hꢀ
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6S, Hꢀ6'S), 5.05 (d, 2H, J = 3.7 Hz, Hꢀ1, Hꢀ1'), 5.35 (t, 2H, J = 4.8 Hz, CH=CH); C NMR δ
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175.44, 175.41 (2 C=O), 130.9, 130.8 (CH=CH), 95.1 (Cꢀ1, Cꢀ1'), 74.5, 73.1 (Cꢀ2, Cꢀ2', Cꢀ3, Cꢀ3'),
71.9, 71.5 (Cꢀ4, Cꢀ4', Cꢀ5, Cꢀ5'), 64.4 (Cꢀ6, Cꢀ6'), 40.3 (CH), 35.1, 33.1 (COCH2, COCH2CH2),
31.1, 30.9, 30.8, 30.8, 30.6, 30.5, 30.4, 30.3, 30.2, 29.2, 28.6, 28.2, 26.1 (CH2), 23.8, 23.1
(CH2CH3), 14.9 (Me); HR ESI MS m/z calc for C45H82NaO13: 853.5648; found: 853.5626.
6ꢀOꢀ(12ꢀMethyltetradecanoyl)ꢀ6'ꢀOꢀoleoylꢀα,αꢀtrehalose (5c). The title compound was
synthesized using procedure B above under the conditions of Table 2, entry 6 with 6ꢀOꢀoleoylꢀα,αꢀ
trehalose (100 mg, 0.17 mmol) and 12ꢀmethyltetradecanoic acid (44 mg, 0.18 mmol) and was
1
obtained as a gummy solid (108 mg, 79%): RF 0.51 [20% MeOH in EtOAcꢀDCM(1:1), v/v]; H
NMR (DMSO) δ 0.80 ꢀ 0.90 (m, 9H, 3 x Me), 1.05 ꢀ 1.12 (m, 2H, CH2 ), 1.16 ꢀ1.18 (m, 2H, CH2),
1.20 ꢀ 1.35 (m, 37H, CHCH2CH3, 19 x CH2), 1.48 ꢀ 1.51 (m, 2H, CH2CH3), 1.95 ꢀ 1.99 (m, 2H,
CH2CHCH), 2.26 (t, 4H, J = 7.0 Hz, CH2CO), 3.10 ꢀ 3.14 (m, 2H, Hꢀ4, Hꢀ4'), 3.22 ꢀ 3.26 (m, 2H,
Hꢀ2, Hꢀ2'), 3.52 ꢀ 3.57 (m, 2H, Hꢀ3, Hꢀ3'), 3.86 ꢀ 3.90 (m, 2H, Hꢀ5, Hꢀ5'), 4.02 (dd, 2H, J =5.7 Hz,
11.7 Hz, Hꢀ6R, Hꢀ6'R), 4.22 (m, 2H, Hꢀ6S, Hꢀ6'S), 4.70 (d, 2H, J = 5.5 Hz, OH), 4.82 (d, 2H, J
=3.6 Hz, Hꢀ1,Hꢀ1'), 4.97 (d, 2H, J = 5.5 Hz, OH), 5.07 (d, 2H, J = 5.5 Hz, OH) 5.30 (t, 2H, J = 5
Hz, CH=CH); 13C NMR δ 172.70, 172.68, (C=O), 129.6 (CH=CH), 93.3 ( Cꢀ1, Cꢀ1'), 72.7,
71.4,71.3, 70.1, 70.0, 69.7 (Cꢀ2, Cꢀ2', Cꢀ3, Cꢀ3',Cꢀ4, Cꢀ4', Cꢀ5, Cꢀ5'), 63.1 (Cꢀ6, Cꢀ6'), 36.0, 33.8,
33.6, 31.3 (COCH2, COCH2CH2), 29.4, 29.0, 28.9, 28.86, 28.74, 28.70, 28.6, 28.50, 28.46, 26.6,
26.5 (CH, CH2), 24.3, 22.1 (CH2CH3), 13.9, 11.2 (Me); HR ESI MS m/z calc for C45H82NaO13:
853.5648; found: 853.5620.
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Acknowledgment. We thank Natural Sciences and Engineering Research Council of Canada
(NSERC) for support and NMRꢀ3 for NMR time.
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Supporting Information Available General experimental procedures and 1H and 13C NMR spectra
for all compounds prepared. This information is available free of charge via the Internet at
http://pubs.acs.org.
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