Synthesis of trehalose-based compounds and their inhibitory activities against Mycobacterium smegmatis

Article abstract of DOI:10.1016/j.bmc.2004.09.033

The synthesis of a library of trehalose-based compounds has been accomplished, and their activities against Mycobacterium smegmatis have been determined. A preliminary structure-activity relationship (SAR) is reported. Despite not having a potent lead, on

Full text of DOI:10.1016/j.bmc.2004.09.033

Bioorganic & Medicinal Chemistry 12 (2004) 6397–6413
Synthesis of trehalose-based compounds and their
inhibitory activities against Mycobacterium smegmatis
Jinhua Wang,a Bryan Elchert,a Yu Hui,a Jon Y. Takemoto,b Mekki Bensaci,b
John Wennergren,a Huiwen Chang,a Ravi Raia and Cheng-Wei Tom Changa,*
aDepartment of Chemistry and Biochemistry, Utah State University, 0300 Old Main Hill, Logan, UT 84322-0300, USA
bDepartment of Biology, Utah State University, 0300 Old Main Hill, Logan, UT 84322-0300, USA
Received 20 July 2004; revised 17 September 2004; accepted 17 September 2004
Available online 13 October 2004
Abstract—The synthesis of a library of trehalose-based compounds has been accomplished, and their activities against Mycobacte-
rium smegmatis have been determined. A preliminary structure–activity relationship (SAR) is reported. Despite not having a potent
lead, one of the trehalose derivatives displays strong activity when applied with isoniazid (INH), which is known to have low ster-
ilizing activity. The bacteriocidal nature of our compounds against Mycobacterium may be significant for the development of new
therapies against tuberculosis.
Ó 2004 Elsevier Ltd. All rights reserved.
ag 85
protein
complex
1. Introduction
HO
O
O
HO
O
O
O
OH
OH
O
OH
O
R
O
OH
HO
HO
OH
Tuberculosis (TB) is still the leading cause of death
from a single infectious agent.1,2 Once curable, the
emergence of multi-drug resistant variants of M. tuber-
culosis (MDR-TB) have challenged normal therapeutic
practices and increased mortality rates worldwide.3
The high lipophilicity of the bacterial cell wall of
Mycobacterium, which lowers the permeability of anti-
tubercular agents and contributes to significant drug
resistance, imposes great challenges in developing new
drugs against TB.4–6
HO
HO
OH
OH
HO
OH
O
O
trehalose
R
+
O
trehalose dimycolate (TDM)
mycolylic acid
R
Scheme 1.
sents a desirable target for the development of new anti-
tubercular agents. More importantly, ag85 is a new
target, and resistance related to ag85 modification has
not been observed so far.
Fortunately, a potential target for the development of
new antitubercular drugs has been revealed. Recent
advances in the studies of the lipid envelope of TB have
uncovered a unique structural component, 6,60-dimyco-
lyltrehalose (TDM), which forms the outmost layer of
the bacterial cell wall of TB.5,6 Three homologous pro-
teins (ag85 A, B, and C), which possess mycolyltransf-
erase activity, are responsible for the biosynthesis of
TDM (Scheme 1).7,8 Damage to the cellwall, due to
the inhibition of ag85, has been shown to increase the
efficacy of various antibiotics.7,9 Therefore, ag85 repre-
Inhibitors derived from trehalose that mimic the scaffold
of TDM have been reported with modest to excellent
activity against TB.10 Based on the proposed mechanism
of action of ag85, we designed and synthesized a second
generation of trehalose-based antitubercular agents.11
These agents contain a characteristic feature of long
hydrocarbon chains and functionalities that may serve
as acyltransferase inhibitors to target ag85 protein com-
plex. Herein, we wish to report the synthesis and prelimi-
nary antibacterial studies of these trehalose-based
compounds against M. smegmatis, a nonpathogenic
surrogate of M. tuberculosis.
Keywords: Tuberculosis; Trehalose; Ag85 protein complex.
*
Corresponding author. Tel.: +1 435 797 3545; fax: +1 435 797
3390; e-mail: chang@cc.usu.edu
0968-0896/$ - see front matter Ó 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2004.09.033

6398
J. Wang et al. / Bioorg. Med. Chem. 12 (2004) 6397–6413
2. Results
BnO
O
O
BnO
O
OBn
OR2
O
OBn
BnO
R1O
O
OBn
a
Our designed compounds are shown in Figure 1. These
molecules contain a trehalose core with mono- or di-
substituted hydrocarbon chains, varying in chain length,
at the C-4 or C-6 position. Amide and urea linkages that
are commonly found in the designs of inhibitors against
proteases, esterases, and peptidases are employed as the
linkages for attaching these hydrocarbon chains. A pre-
liminary goal is to identify optimal functionalities and
chain length. For compounds like YH023 the amide
linkage is constructed in a ꢀreversedꢁ fashion compared
to the linkage found in TDM.
O
BnO
BnO
HO
OBn
OBn
OH
BnO
2, R1 = R2 = C8H17 64%
OBn
1
3, R1 =H, R2 = C8H17 41%
HO
O
O
OH
OR2
O
b
HO
R1O
OH
HO
OH
YH008, R1 = R2 = C8H17 84%
YH018, R1 =H, R2 = C8H17 94%
BnO
O
BnO
O
O
OBn
OR
O
O
OBn
OH
O
a
Synthesis of trehalose-based compounds with an ether
linkage starts from the known compounds, 1,11 4,11
and 6,12 bearing free either 4-OH or 6-OH groups
(Scheme 2). The hydrocarbon chains are attached via
alkylation using the corresponding alkyl halide. By con-
trolling the number of equivalents of alkyl halide, mono-
and di-substituted alkyl trehalose derivatives can be
obtained in one pot. Hydrogenation using Pd(OH)2/C
as a catalyst yields the final products.
OBn
RO
BnO
OBn
HO
BnO
4
5, R = C8H17 67%
HO
O
OH
OR
O
b
O
OH
RO
HO
R = C8H17 72%
YH011
BnO
O
O
BnO
O
O
OBn
O
OBn
R1O
O
Synthesis of trehalose-based compounds bearing an
amide linkage with the C-terminal on the trehalose
begins from 6 (Scheme 3). After Swern oxidation,13
the resulting aldehyde is divided into two parts for sep-
OBn
a
HO
BnO
OBn
OR2
OBn
BnO
BnO
OBn
OH
BnO
7, R1 = R2 = C8H17 16%
6
8, R1 =H, R2 = C8H17 29%
HO
O
O
OH
O
b
R1O
HO
OH
HO
O
HO
O
O
O
OH
OR2
HO
OH
OR
OH
OR2
O
17
O
HO
OH
OH
R1O
HO
YH011,
RO
HO
YH019, R1 = R2 = C8H17 72%
OH
YH020, R1 =H, R2 = C8H17 63%
R = C H
8
YH008, R1 = R2 = C8H17
YH018, R1 =H, R2 = C8H17
Scheme 2. Reagents: (a) C8H17Br, NaH, TBAI, THF; (b) H2,
Pd(OH)2/C, MeOH.
O
HO
O
HO
O
O
H
OH
OH
OH
O
O
O
R1O
HO
R
N
HO
OH
OH
OH
OR2
HO
HO
O
N
R
YH012, R=C8H17
YH017, R=C12H25
YH019,
YH020,
R
R
1 = R2 = C8H17
H
1 =H, R2 = C8H17
arate routes. In one branch, the aldehyde is further oxi-
dized into the carboxylic acid, which is then coupled
with alkyl amine and L-alanine ethyl ester using (COCl)2
and EDC, respectively. Hydrogenation using Pd(OH)2/
C as a catalyst gives the final amides. In the other route,
treatment of the aldehyde with methyl diethylphospho-
noacetate provides 14 with a two-carbon extension at
C-6 (Scheme 4). Transamidation14 of 14 using AlMe3
and the corresponding amines or hydrazine, followed
by hydrogenation, yields the desired trehalose-based
derivatives with a reversed amide linkage, with respect
to the corresponding ester linkage found in TDM.
O
HO
HO
O
O
O
OH
O
OH
OH
O
O
O
N
H
EtOOC
OH
OH
RN
OH
HN
HO
HO
HO
HO
H
O
YH024
O
H
COOEt
YH023, R = C8H17
YH027, R = C12H25
YH028, R = C4H9
YH029, R = C5H11
YH030, R = C6H13
YH031, R = C7H15
YH032, R = C9H19
YH033, R = C10H21
N
R
HO
O
O
O
OH
OH
O
C8H17HN HN
HO
OH
HO
O
HN
YH034, R = CH3CH2O(CH2)3
YH035, R = CH3(CH2)3O(CH2)3
NHC8H17
YH026
HO
O
O
H
N
O
OH
OH
O
HO
O
O
O
C7H15
OH
OH
OH
N
H
O
C9H19
N
H
HO
HO
OH
O
Synthesis of trehalose-based compounds bearing an
amide linkage with the N-terminal on the trehalose
and urea linkage also begins from 6 (Scheme 5). The
6-amino groups are installed via triflation and azide sub-
stitution, followed by a Staudinger reaction. Treatment
of the amine, 26, with triphosgene yields an isocyanide,
which is then reacted with octyl amine to generate the
urea adduct. In another route, compound 26 is coupled
with decanoyl chloride, leading to the desired compound
with an amide linkage. Both of the benzyl-protected
HO
HO
O
O
HN
YH025
YH036
H N
O
C9H19
N H
C7H15
HO
O
HO
O
O
O
C7H15NH
OH
OH
O
O
H2N
HO
O
CH2
N
H
OH
OH
YH041
OH
HN
OH
O
HO
HO
HO
CH2
YH037
O
NHC7H15
NHC8H17
Figure 1. Structure of the second generation inhibitors.

J. Wang et al. / Bioorg. Med. Chem. 12 (2004) 6397–6413
6399
O
BnO
O
BnO
O
O
O
OBn
OBn
O
O
a
HO
HO
BnO
OBn
OBn
79%
OBn
OH
OBn
BnO
BnO
BnO
O
O
OH
6
9
b
25%
c
O
BnO
O
O
BnO
O
H
OBn
OBn
O
O
O
OBn
O
R
R
N
OBn
N
H
BnO
BnO
EtOOC
OBn
O
N
R
OBn
BnO
BnO
10, R=C8H17, 33%
HN
12
H
11, R=C12H25, 37%
COOEt
d
43%
d
O
HO
O
O
HO
O
O
H
OH
O
OH
O
O
N
H
N
OH
EtOOC
OH
OH
HO
HO
OH
HN
HO
O
HO
N
H
R
O
YH024
YH012, R=C8H17, 99%
YH017, R=C12H25, 80%
COOEt
Scheme 3. Reagents: (a) (1) (COCl)2, DMSO, DIPEA, (2) NaClO2, H2O2; (b) (1) (COCl)2, (2) C8H17NH2 or C12H25NH2; (c) L-alanine ethylester,
EDC, pyr.; (d) H2, Pd(OH)2/C, MeOH.
BnO
O
BnO
O
O
O
OBn
OBn
O
OBn
O
O
a
b
OBn
MeO
HO
BnO
OBn
BnO
BnO
77%
OBn
OH
BnO
14
6
O
MeO
HO
BnO
O
O
O
O
O
OH
O
OBn
OBn
O
O
c
80 - 90%
OH
OBn
RN
OH
RN
H
HO
HO
BnO
BnO
H
O
YH023, R = C8H17
YH027, R = C12H25
YH028, R = C4H9
YH029, R = C5H11
YH030, R = C6H13
YH031, R = C7H15
YH032, R = C9H19
YH033, R = C10H21
O
H
H
15, R = C8H17, 54%
16, R = C12H25 48%
17, R = C4H9, 45%
N
R
N
R
18, R = C5H11
19, R = C6H13
20, R = C7H15
21, R = C9H19
,
,
,
,
74%
60%
34%
26%
22, R = C10H21, 24%
YH034, R = CH3CH2O(CH2)3
YH035, R = CH3(CH2)3O(CH2)3
23, R = CH3CH2O(CH2)3, 29%
24, R = CH3(CH2)3O(CH2)3, 39%
Scheme 4. Reagents: (a) (1) (COCl)2, DMSO, DIPEA, (2) MeOC(O)CH2P(O)(OEt)2, NaH, THF; (b) alkyl amines, AlMe3, CH2Cl2; (c) H2,
Pd(OH)2/C, MeOH.
compounds are subjected to hydrogenation to furnish
the final products. In an effort to increase the solubility
of trehalose derivatives in aqueous media, we synthe-
sized YH037 with the expectation that the additional
NH group would improve the solubility (Scheme 6).
Coupling of 26 with bromoacetic acid affords 29.
Nucleophilic addition of 29 with heptylamine gives 30,
which is converted into YH037 via hydrogenation.
8). Starting from a known compound 33, one of the
O-6 benzyl groups is selectively removed using Ac2O,
TMSOTf followed by base hydrolysis. Azido substitu-
tion at the O-6 position provided 35, which is converted
to 36 with the same conditions. The final product,
YH041 is synthesized from the identical procedures for
the preparation of YH023. YH041 is found to be soluble
in water.
We also designed YH036 with a diacylhydrazine func-
tionality with the expectation that the added –CONH–
group will increase both antibacterial activity and solu-
bility (Scheme 7). YH036 is synthesized from 14 through
hydrolysis, coupling with octanoic hydrazide, and
hydrogenation. We also constructed a trehalose deriva-
tive, YH041 with a mono-hydrocarbon chain (Scheme
After completion of the synthesis, the trehalose deriva-
tives are assayed against M. smegmatis (ATCC 14468),
using isoniazid (INH) as the positive control.15 Since
M. tuberculosis is pathogenic and not easily manipulated
experimentally, M. smegmatis is typically used to
investigate issues related to the biology and pathogene-
sis of the former. Although associated with disease,

6400
J. Wang et al. / Bioorg. Med. Chem. 12 (2004) 6397–6413
Table 1. Inhibition zones of trehalose derivatives against M.
BnO
O
BnO
O
O
O
a
OBn
OBn
O
smegmatis
O
N3
HO
OBn
OBn
74%
OBn
N3
OBn
OH
BnO
Compounda
Inhibition zone (mm)
BnO
BnO
BnO
25
6
INH
18
6
Inactive
BnO
O
YH008
YH011
YH012
YH017
YH018
YH019
YH020
YH023
YH024
YH025
YH026
YH027
YH028
YH029
YH030
YH031
YH032
YH033
YH034
YH035
YH036
YH037
YH041
OBn
O
b
O
OBn
H2N
78%
OBn
NH2
10
6
BnO
BnO
c
26
49%
d
5
11
91%
BnO
O
O
O
BnO
O
O
Inactive
20
OBn
OBn
O
O
OBn
OBn
C9H19
N
H
O
OBn
C8H17HN
HN
BnO
OBn
BnO
BnO
5
10
O
BnO
28
HN
O
NHC8H17
27
HN
C9H19
e
99%
8
e
78%
Inactive
Inactive
Inactive
Inactive
8
HO
O
O
O
HO
O
OH
OH
O
O
C9H19
OH
OH
N
H
O
O
OH
C8H17HN
HN
HO
OH
HO
HO
O
HO
HN
O
YH025
HN
YH026
C9H19
11
6
NHC8H17
Inactive
Inactive
12
Scheme 5. Reagents: (a) (1) Tf2O, pyr., CH2Cl2, (2) NaN3, DMF; (b)
PMe3, THF, NaOH; (c) triphosgene, Et3N, C8H17NH2; (d)
C9H19COCl, CH2Cl2; (e) H2, Pd(OH)2/C, MeOH.
19
Inactive
a Concentration used: INH: 0.25mg/mL, trehalose derivatives: 10mg/
mL.
BnO
O
BnO
O
O
O
O
OBn
OBn
O
M. smegmatis is not deemed an important human
pathogen.16 It is more amenable to experimentation
since unlike M. tuberculosis it is a fast grower and geneti-
cally more tractable.17 Most importantly, M. smegmatis
shares several clinically important properties that charac-
terize M. tuberculosis, including similar resistance to
certain macrolide drugs.18 Due to the poor solubility
of our trehalose derivatives, DMSO/water solutions
were used for the diffusion assay. The antibacterial
activity of the trehalose derivatives is evaluated by the
standard diffusion assay. The inhibition zones are sum-
marized in Table 1.19
O
a
BrH2C
N
BnO
H2N
BnO
OBn
OBn
H
OBn
HN
OBn
NH2
74%
BnO
BnO
b
26
29
CH2Br
O
BnO
O
O
c
C7H15NH
OBn
O
O
CH2
N
75%
99%
OBn
H
OBn
O
BnO
BnO
CH2
HN
30
NHC7H15
HO
O
O
CH2
O
C7H15NH
OH
O
N
OH
H
OH
O
HOHO
CH2
HN
YH037
3. Discussion
NHC7H15
The attempts to determine standard MIC values in liq-
uid broth were hampered by the poor water-solubility.
Scheme 6. Reagents: (a) BrCH2CO2H, DMF; DIC; (b) C7H15NH2,
Cs2CO3, DMF; (c) H2, Pd/C, MeOH.
BnO
O
BnO
O
O
OBn
OBn
b
O
O
OBn
OBn
O
O
a
O
OBn
HO
OBn
41%
MeO
BnO
BnO
36%
BnO
BnO
31
14
O
O
HO
MeO
HO
O
H
N
BnO
O
O
O
OH
OH
O
H
N
O
O
OBn
OBn
O
C7H15
OH
N
H
c
C7H15
HO
OBn
HO
N
O
BnO
BnO
O
H
O
99%
O
H N
O
32
YH036
H N
O
N H
C7H15
N H
C7H15
Scheme 7. Reagents: (a) LiOH–H2O, THF, H2O; (b) H2NNHC(O)C7H15; (c) H2, Pd(OH)2/C, MeOH.

J. Wang et al. / Bioorg. Med. Chem. 12 (2004) 6397–6413
6401
BnO
O
Nevertheless, the MIC for the most active compound,
BnO
O
O
O
b
a
OBn
OBn
OBn
OBn
O
YH023 is estimated to be >256lM. We have also de-
rived the structure–activity relationship of the trehalose
derivatives:
O
HO
84%
OBn
BnO
BnO
OBn
50%
BnO
BnO
BnO
34
33
OBn
OBn
BnO
BnO
O
O
1. Compounds with di-hydrocarbon chain have similar
or better activities than compounds with mono-
hydrocarbon chain.20
OBn
OBn
a
O
c
O
N3
N3
O
O
OBn
OBn
50%
99%
OBn
OH
OBn
OBn
BnO
BnO
BnO
BnO
35
36
2. The activity trends among functionalities:
BnO
BnO
O
O
O
OBn
OBn
OBn
O
O
reversed acyl ꢀ a-alkylaminoacetyl > acyl ꢀ urea >
N3
N3
BnO
O
d
OBn
OBn
ether > compounds without hydrocarbon chain
OBn
O
BnO
BnO
BnO
45%
38
37
O
3. The optimal chain length is around C8 or C9.
4. Incorporation of oxygen atoms within the long
hydrocarbon chain cannot be tolerated.
OMe
NHC8H17
HO
O
OH
O
H2N
HO
O
Despite significant progress, the mechanism of INH ac-
tion is not fully understood. It is generally proposed that
INH is a prodrug that can be rapidly transported into
actively growing bacteria then activated via the action
of catalase/peroxidase.21 Several enzymes have been re-
ported as the targets of activated INH, including en-
zymes for the biosynthesis of mycolic acid,22 catalase/
peroxidase involved in the activation of INH,23 and
alkyl hydroperoxide reductase.24 While highly active
against M. tuberculosis, INH is noted for its rapid steri-
lizing activity but lack of effectiveness during TB
relapse.25 The later leads to the requirement for long-
term INH chemotherapy, and, hence, the increased
chance of treatment failure. Unlike INH, the treha-
lose-based compound with reversed acyl linkage,
YH023, is bacteriocidal.
e
OH
OH
O
HO
52%
YH041
NHC8H17
Scheme 8. Reagents: (a) (1) Ac2O, TMSOTf, (2) NaOMe, MeOH; (b)
(1) Tf2O, CH2Cl2, pry., (2) NaN3, DMF; (c) (1) (COCl)2, DMSO,
DIPEA, (2) MeOC(O)CH2P(O)(OEt)2, NaH, THF; (d) C8H17NH2,
AIMe3, CH2Cl2; (e) H2, Pd(OH)2/C, MeOH.
combined and employed together, we observed that
the growth inhibitory zones of M. smegmatis were
slightly larger than with INH alone at the same concen-
trations. Furthermore, with the YH023–INH combina-
tion, no return of growth was observed over a period
of 2weeks in the initially inhibited zones (Fig. 2). In con-
trast, significant growth occurred in the inhibitory zones
with INH alone. This result suggests that YH023 is
synergistic with INH against M. smegmatis. This finding
is potentially significant in shortening the timeline of TB
treatment, leading to higher rates of success and lower
cost in the battle against this disease.
INH has been found to induce the over-expression of
ag85 protein complex,26 and transient high-level resist-
ance in M. tuberculosis.27 On the other hand, it has been
reasoned that the trehalose derivatives can act as inhib-
itors of ag85 protein complex.10 Therefore, we evaluate
the synergistic or additive effect between INH and the
lead compound YH023. When INH and YH023 were
4. Conclusion
In summary, we have produced a new class of anti-TB
compounds that may target the ag85 protein complex,
which is a new target for the development of anti-TB
agents. The observation that our compounds are bacte-
riocidal against Mycobacterium suggests that they may
be developed to address resistance problems common
to the use of current anti-TB drugs. We are currently
synthesizing trehalose derivatives with the goal of
increasing their solubility in aqueous media and the anti-
bacterial activity of these compounds.
5. Experimental
Proton magnetic resonance spectra were recorded using
a Jeol 270 or Bruker 400 spectrometers. Chemical shifts
were reported as parts per million (ppm) downfield from
tetramethylsilane in d unit, and coupling constants were
given in cycles per second (Hz). Splitting patterns were
designed as s, singlet; d, doublet; t, triplet; q, quartet;
Figure 2. Assay of YH023 and INH against M. smegmatis in water/
DMSO solution. A: INH (250lg/mL); B: INH (250lg/mL) + YH023
(10mg/mL); C: DMSO; D: YH023 (10mg/mL).

6402
J. Wang et al. / Bioorg. Med. Chem. 12 (2004) 6397–6413
m, multiplet. 13C spectra were obtained using the Jeol
270 spectrometer at 68MHz, and were reported in
ppm with center line of the triplet at 77.0ppm for chlo-
roform-d. Routine 13C NMR spectra were fully decou-
pled by broad-band waltz decoupling. All NMR
spectra were recorded at ambient temperature unless
otherwise noted. Low-resolution fast-atom bombard-
ment (LRFAB) and high-resolution fast-atom bom-
bardment (HRFAB) were provided by the Mass
Spectrometry Facilities, University of California,
Riverside.
removal of solvent, the crude product was diluted with
Et2O. The organic solution was washed with 1N
HCl(aq), water, saturated NaHCO3(aq), and brine, then
dried over Na2SO4(s). After removal of the solvent
followed by purification with a gradient column
chromatography (hexane–EtOAc = 100:0 to 60:40), the
product was obtained as white powder (0.070g,
0.070mmol, 41%). 1H NMR (270MHz, CDCl3) d
7.10–7.50 (m, 30H), 5.23 (d, J = 3.6Hz, 1H, H-1), 5.20
(d, J = 3.6Hz, 1H, H-10), 5.02 (d, J = 11.2Hz, 1H,
PhCH2O), 4.95 (d, J = 10.9Hz, 1H, PhCH2O), 4.83 (d,
J = 10.9Hz, 1H, PhCH2O), 4.80 (d, J = 11.6Hz, 1H,
PhCH2O), 4.70 (d, J = 11.9Hz, 1H, PhCH2O), 4.66 (s,
2H, PhCH2O), 4.64 (d, J = 11.9Hz, 1H, PhCH2O),
4.4–4.6 (m, 4H), 4.1–4.2 (m, 2H), 3.9–4.0 (m, 2H), 3.7–
3.8 (m, 2H), 3.3–3.6 (m, 2H), 1.00–2.00 (m, 12H), 0.89
(t, J = 6.3Hz, 3H, CH2CH3); 13C NMR (68MHz,
CDCl3) d 139.0 (s), 138.9 (s), 138.3 (s), 138.2 (s), 138.0
(s), 137.9 (s), 128.6 (s), 128.4 (s), 128.1 (s), 128.0 (s),
127.9 (s), 127.7 (s), 127.7 (s), 127.6 (s), 127.6 (s), 127.5
(s), 94.5 (s, C-1), 94.3 (s, C-10), 81.8 (s), 81.1 (s), 79.2
(s), 79.1 (s), 77.8 (s), 75.6 (s), 75.3 (s), 73.7 (s), 73.6 (s),
73.3 (s), 72.9 (s), 72.5 (s), 70.9 (s, 2C), 70.6 (s), 69.3
(s), 68.3 (s), 32.0 (s), 30.6 (s), 29.6 (s), 29.4 (s), 26.3 (s),
22.8 (s), 14.2 (s); MALDI Calcd for C62H74O11Na
([M+Na]+) 1017.5123; measure 1017.5125.
Chemical reagents and starting materials were pur-
chased from Aldrich Chemical Co. or Acros Chemical
Co. and were used without purification unless otherwise
noted. Dichloromethane was distilled over CaH2. Tetra-
hydrofuran, triethylamine, diisopropylethylamine dried
˚
and stored with activated 4A molecular sieves under
nitrogen. Pyridine was dried over CaH2 and kept under
nitrogen.
5.1. 2,20,3,30,6,60-Hexa-O-benzyl-4,40-di-O-octyl-
a-D-trehalose (2)
To a solution of 1 (0.10g, 0.11mmol) dissolved in 3mL
of anhydrous THF at 0°C, TBAI (0.02g, catalytic
amount), NaH (0.02g, 0.72mmol), and 1-octyl bromide
(0.063mL, 0.36mmol) were added. The reaction was
stirred for 12h, allowing the temperature to be warmed
up to room temperature. After completion of the reac-
tion, the reaction was quenched with few drops of
MeOH. After removal of solvent, the crude product
was diluted with Et2O. The organic solution was washed
with 1N HCl(aq), water, saturated NaHCO3(aq), and
brine, then dried over Na2SO4(s). After removal of the
solvent followed by purification with a gradient column
chromatography (hexane–EtOAc = 100:0 to 60:40), the
product was obtained as white powder (0.08g,
General procedure for hydrogenation: The starting mate-
rial was added with catalytic amount of Pd(OH)2/C
(20% Degussa type) or Pd/C (10%) and 5mL of degassed
MeOH or MeOH–EtOAc (1:1). After being further de-
gassed, the reaction mixture was stirred at room temper-
ature under atmospheric H2 pressure. After being stirred
for 1day, the reaction mixture was filtered through Cel-
ite. The residue was washed with MeOH, and the com-
bined solution was concentrated, affording pure final
product.
1
0.07mmol, 64%). H NMR (270MHz, CDCl3) d 7.10–
5.3. 4,40-Di-O-octyl-a-D-trehalose (YH008)
7.50 (m, 30H), 5.20 (d, J = 3.3Hz, 2H, H-1), 4.95 (d,
J = 10.9Hz, 2H, PhCH2O), 4.85 (d, J = 10.9Hz, 2H,
PhCH2O), 4.67 (s, 4H, PhCH2O), 4.55 (d, J = 11.8Hz,
2H, PhCH2O), 4.40 (d, J = 11.8Hz, 2H, PhCH2O),
4.05–4.10 (m, 2H), 3.93 (t, J = 9.6Hz, 2H), 3.70–3.82
(m, 2H), 3.30–3.60 (m, 10H), 1.10–1.60 (m, 24H), 0.87
(t, J = 6.9Hz, 6H, CH2CH3); 13C NMR (68MHz,
CDCl3) d 139.1 (s), 138.4 (s), 138.0 (s), 128.4 (s), 128.0
(s), 127.7 (s), 127.5 (s), 94.6 (s, C-1), 81.8 (s), 79.4 (s),
77.8 (s), 75.6 (s), 73.5 (s), 73.3 (s), 72.8 (s), 70.8
(s), 68.3 (s), 32.0 (s), 30.7 (s), 29.7 (s), 29.4 (s), 26.3 (s),
22.8 (s), 14.2 (s); MALDI Calcd for C70H90O11Na
([M+Na]+) 1129.6375; measure 1129.6412.
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 4.82 (d, J = 3.3Hz, 2H,
H-1), 3.30–4.00 (m, 12H), 3.22 (dd, J = 9.8Hz,
J = 3.3Hz, 2H, H-2), 3.04 (t, J = 9.8Hz, 2H, H-4),
1.00–1.60 (m, 24H), 0.87 (t, J = 6.9Hz, 6H, CH2CH3);
13C NMR (68MHz, CD3OD) d 93.7 (s, C-1), 78.4 (s),
73.2 (s), 72.3 (s), 72.2 (s), 71.8 (s), 60.8 (s), 31.8
(s), 30.5 (s), 29.5 (s), 29.3 (s), 26.2 (s), 22.7 (s), 14.5 (s);
LRFAB m/e 589 ([M+Na]+); HRFAB Calcd for
C28H54O11Na ([M+Na]+) m/e 589.3564; measure m/e
589.3540.
5.4. 4-O-Octyl-a-D-trehalose (YH018)
5.2. 2,20,3,30,6,60-Hexa-O-benzyl-4-O-octyl-a-D-trehalose
(3)
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 5.09 (d, J = 3.3Hz, 1H,
H-1), 5.08 (d, J = 3.3Hz, 1H, H-10), 3.00–4.00 (m, 14H),
1.00–2.00 (m, 12H), 0.88 (t, J = 5.9Hz, 3H, CH2CH3);
13C NMR (68Hz, CD3OD) d 93.7 (s, C-1), 93.6 (s,
C-10), 78.3 (s), 73.3 (s), 73.2 (s), 72.6 (s), 72.4 (s), 72.1
(s), 71.9 (s), 71.6 (s), 70.6 (s), 61.3 (s), 60.8 (s), 31.7 (s),
30.1 (s), 29.3 (s), 29.1 (s), 25.9 (s), 23.4 (s), 13.1 (s);
LRFAB m/e 477 ([M+Na]+); HRFAB Calcd for
To a solution of 1 (0.15g, 0.17mmol) dissolved in 3mL
of THF at 0°C, TBAI (0.01g, catalytic amount), NaH
(0.012g, 0.51mmol), and 1-octyl bromide (0.050mL,
0.26mmol) were added. The reaction was stirred for
12h, allowing the temperature to be warmed up to room
temperature. After completion of the reaction, the reac-
tion was quenched with few drops of MeOH. After

J. Wang et al. / Bioorg. Med. Chem. 12 (2004) 6397–6413
6403
C20H38O11Na ([M+Na]+) m/e 477.2312; measure m/e
477.2305.
chromatography. Pure products 7 and 8 were collected
(0.09g, 0.08mmol, 16% and 0.15g, 0.15mmol, 29%,
respectively).
5.5. 2,20,3,30-Tetra-O-benzyl-4,40-di-O-octyl-6,60-dide-
oxy-a-D-trehalose (5)
5.8. 2,20,3,30,4,40-Hexa-O-benzyl-6,60-di-O-octyl-a-D-tre-
halose (7)
To a solution of 4 (0.10g, 0.15mmol) dissolved in 3mL
of anhydrous THF at 0°C, TBAI (0.020g, catalytic
amount), NaH (0.023g, 0.96mmol), and 1-octyl bro-
mide (0.080mL, 0.48mmol) were added. The reaction
was stirred for 12h, allowing the temperature to be
warmed up to room temperature. After completion of
the reaction, the reaction was quenched with few drops
of MeOH. After removal of solvent, the crude product
was diluted with Et2O. The organic solution was washed
with 1N HCl(aq), water, saturated NaHCO3(aq), and
brine, then dried over Na2SO4(s). After removal of the
solvent followed by purification with a gradient column
chromatography (hexane–EtOAc = 100:0 to 60:40), the
product was obtained as white powder (0.090g,
1H NMR (270MHz, CDCl3) d 7.3–7.4 (m, 30H), 5.21 (d,
J = 3.6Hz, H-1, 2H), 4.99 (d, J = 10.9Hz, 2H,
PhCH2O), 4.86 (d, J = 10.9Hz, 4H, PhCH2O), 4.68 (s,
4H, PhCH2O), 4.59 (d, J = 10.9Hz, 2H, PhCH2O),
4.15 (d, J = 10.2Hz, 2H), 4.03 (dd, J = 9.6Hz,
J = 9.2Hz, 2H), 3.68 (dd, J = 9.9Hz, J = 9.2Hz, 2H),
3.58 (dd, J = 9.6Hz, J = 3.6Hz, 2H), 3.2–3.5 (m, 8H),
1.5–1.6 (m, 4H), 1.2–1.3 (m, 20H), 0.86 (t, J = 6.9Hz,
6H); 13C NMR (68MHz, CDCl3) d 139.0 (s), 138.7 (s),
138.6 (s), 128.5 (s), 128.42 (s), 128.41 (s), 128.0(s),
127.96 (s), 127.8 (s), 127.6 (s), 94.6 (s, C-1), 81.9
(s), 79.5 (s), 77.8 (s), 75.7 (s), 75.2 (s), 72.7 (s), 71.9 (s),
70.7 (s), 69.0 (s), 31.9 (s), 29.7 (s), 29.6 (s), 29.3 (s),
26.3 (s), 22.7 (s), 14.2 (s); MALDI Calcd for
C70H90O11Na ([M+Na]+) m/e 1129.6375; measure m/e
1129.6340.
1
0.10mmol, 67%). H NMR (270MHz, CDCl3) d 7.10–
7.50 (m, 20H), 5.09 (d, J = 3.6Hz, 2H, H-1), 4.96 (d,
J = 10.9Hz, 2H, PhCH2O), 4.85 (d, J = 10.9Hz, 2H,
PhCH2O), 4.70 (s, 4H, PhCH2O), 4.06–4.20 (m, 2H),
3.93 (t, J = 9.2Hz, 2H), 3.83 (dt, J = 9.2Hz, J =
6.6Hz, 2H), 3.50–3.61 (m, 2H), 3.51 (dd, J = 9.2Hz,
J = 3.6Hz, 2H, H-2), 2.93 (t, J = 9.2Hz, 2H), 1.20–
1.65 (m, 24H), 1.14 (d, J = 6.6Hz, 6H, H-6), 0.89 (t,
J = 6.3Hz, 6H, CH2CH3); 13C NMR (68MHz, CDCl3)
d 139.1 (s), 138.4 (s), 128.4 (s), 128.0 (s), 127.6 (s), 94.2
(s, C-1), 84.4 (s), 81.4 (s), 79.8 (s), 75.7 (s), 73.6 (s),
72.8 (s), 67.4 (s), 32.0 (s), 30.6 (s), 29.6 (s), 29.4 (s),
26.3 (s), 22.8 (s), 17.9 (s), 14.2 (s); MALDI Calcd for
C56H78O9Na ([M+Na]+) 917.5538; measure 917.5559.
5.9. 2,20,3,30,4,40-Hexa-O-benzyl-6-O-octyl-a-D-trehalose
(8)
1H NMR (270MHz, CDCl3) d 7.3–7.4 (m, 30H), 5.21 (d,
J = 3.9Hz, 1H), 5.19 (d, J = 3.9Hz, 1H), 5.02 (d,
J = 10.9Hz, 1H, PhCH2O), 5.01 (d, J = 10.9Hz, 1H,
PhCH2O), 4.91 (s, 2H, PhCH2O), 4.87 (s, 2H, PhCH2O),
4.6–4.7 (m, 6H, PhCH2O), 4.0–4.2 (m, 4H), 3.70 (t,
J = 10.8Hz, 1H), 3.2–3.6 (m, 9H), 1.55–1.57 (m, 2H),
1.2–1.3 (m, 10H), 0.88 (t, J = 5.8Hz, 3H); 13C NMR
(68MHz, CDCl3) d 138.98 (s), 138.92 (s), 138.6 (s),
138.4 (s), 138.23 (s), 138.21 (s), 128.6 (s), 128.5 (s),
128.2 (s), 128.0 (s), 127.8 (s), 127.6 (s), 94.4 (s, C-1),
94.2 (s, C-10), 81.9 (s), 81.7 (s), 79.6 (s), 79.5 (s), 77.9
(s), 77.4 (s), 75.7 (s, 2 carbons), 75.2 (s, 2 carbons),
73.0 (s), 72.9 (s), 71.9 (s), 71.3 (s), 70.8 (s), 69.0 (s),
61.6 (s), 31.9 (s), 29.7 (s), 29.6 (s), 29.3 (s), 26.3 (s),
22.8 (s), 14.2 (s); MALDI Calcd for C62H74O11Na
([M+Na]+) 1017.5123; measure 1017.5094.
5.6. 4,40-Di-O-octyl-6,60-dideoxy-a-D-trehalose (YH011)
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 4.93 (d, J = 4.0Hz, 2H,
H-1), 3.82–3.93 (m, 4H), 3.79 (t, J = 9.2Hz, 2H, H-3),
3.56 (dt, J = 9.2Hz, J = 6.6Hz, 2H, H-5), 3.42 (dd,
J = 9.2Hz, J = 4.0Hz, 2H, H-2), 2.79 (t, J = 9.2Hz,
2H, H-4), 1.10–1.60 (m, 24H), 1.20 (d, J = 6.6Hz, 6H,
H-6), 0.89 (t, J = 6.3Hz, 6H, CH2CH3); 13C NMR
(68MHz, CD3OD) d 93.7 (s, C-1), 84.7 (s), 73.2 (s),
72.7 (s), 72.4 (s), 66.9 (s), 31.7 (s), 30.1 (s), 29.3 (s),
29.1 (s), 25.9 (s), 22.4 (s), 17.0 (s), 13.1 (s); LRFAB
m/e 557 ([M+Na]+); HRFAB Calcd for C28H54O9Na
([M+Na]+) m/e 557.3666; measure m/e 557.3652.
5.10. 6,60-Di-O-octyl-a-D-trehalose (YH019)
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 5.08 (d, J = 3.6, 2H, H-
1), 3.89–3.93 (m, 2H), 3.7–3.8 (m, 2H), 3.6–3.7 (m, 4H),
3.4–3.6 (m, 4H), 3.2–3.3 (m, 4H), 1.5–1.6 (m, 4H), 1.2–
1.3 (m, 20H), 0.89 (t, J = 6.9Hz, 6H); 13C NMR
(68MHz, CD3OD) d 93.7 (s, C-1), 73.3 (s), 71.8 (s),
71.4 (s), 71.3 (s), 70.8 (s), 69.8 (s), 31.7 (s), 29.4 (s),
29.3 (s), 29.1 (s), 25.9 (s), 22.4 (s), 13.1 (s); LRFAB
m/e 589 ([M+Na]+); HRFAB Calcd for C28H54O11Na
([M+Na]+) m/e 589.3564; measure m/e 589.3564.
5.7. 2,20,3,30,4,40-Hexa-O-benzyl-6,60-di-O-octyl-a-D-tre-
halose (7) and 2,20,3,30,4,40-hexa-O-benzyl-6-O-octyl-a-
D-trehalose (8)
To a solution of 6 (0.46g, 0.52mmol) in anhydrous
DMF (5mL), NaH (0.03g, 1.04mmol) was slowly added
followed by a catalytic amount of TBAI and octyl bro-
mide (0.14mL, 0.78mmol). The reaction was stirred
overnight at room temperature. After complete con-
sumption of the starting material, analyzed by TLC
(hexane–EtOAc = 65:35), the reaction was quenched
by addition of a few drops of H2O. The solvent was then
removed and the crude residue was purified by column
5.11. 6-O-Octyl-a-D-trehalose (YH020)
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 5.10 (d, J = 3.6Hz, 1H,
H-1), 5.09 (d, J = 3.6Hz, 1H, H-10), 3.88–3.93 (m, 1H),
3.75–3.82 (m, 3H), 3.6–3.7 (m, 2H), 3.4–3.6 (m, 4H),

6404
J. Wang et al. / Bioorg. Med. Chem. 12 (2004) 6397–6413
3.3–3.4 (m, 4H), 1.5–1.6 (m, 2H), 1.30 (m, 10H), 0.89 (t,
J = 6.3Hz, 3H); 13C NMR (68MHz, CD3OD) d 93.74
(s, C-1), 93.66 (s, C-10), 73.3 (s), 73.2 (s), 72.5 (s), 71.9
(s), 71.8 (s), 71.4 (s), 71.3 (s), 70.8 (s), 70.6 (s), 69.8 (s),
61.3 (s), 31.7 (s), 29.4 (s), 29.3 (s), 29.1 (s), 25.9 (s),
22.4 (s), 13.1 (s); LRFAB m/e ([M+Na]+); HRFAB
Calcd for C20H38O11Na ([M+Na]+) m/e 477.2312; meas-
ure m/e 477.2318.
octyl amine (0.073mL, 0.44mmol) at 0°C. The reaction
was stirred for 12h, allowing the temperature to be
warmed up to room temperature. After removal of
the solvent followed by purification with a gradient col-
umn chromatography (hexane–EtOAc = 75:25 to
25:75), the product was obtained as a pale yellowish
oil (0.04g, 0.036mmol, 33%). 1H NMR (270MHz,
CDCl3) d 7.10–7.50 (m, 30H), 5.30 (t, J = 5.6Hz, 2H,
CONH), 5.13 (d, J = 3.3Hz, 2H, H-1), 4.96 (d,
J = 11.2Hz, 2H, PhCH2O), 4.90 (d, J = 11.2Hz, 2H,
PhCH2O), 4.77 (d, J = 12.2Hz, 2H, PhCH2O), 4.76 (d,
J = 10.2Hz, 2H, PhCH2O), 4.65 (d, J = 12.2Hz, 2H,
PhCH2O), 4.54 (d, J = 10.2Hz, 2H, PhCH2O), 4.31 (d,
J = 9.6Hz, 2H, H-5), 4.05 (t, J = 9.6Hz, 2H, H-3),
3.69 (t, J = 9.6Hz, 2H, H-4), 3.62 (dd, J = 9.6Hz,
J = 3.3Hz, 2H, H-2), 2.9–3.0 (m, 4H, CONHCH2),
1.00–2.00 (m, 24H), 0.86 (t, J = 6.6Hz, 6H, CH2CH3);
13C NMR (68MHz, CDCl3) d 168.5 (s, CONH), 138.5
(s), 138.2 (s), 138.0 (s), 128.7 (s), 128.5 (s), 128.4 (s),
128.2 (s), 128.0 (s), 127.8 (s), 127.7 (s), 127.0 (s), 94.8
(s, C-1), 81.4 (s), 80.3 (s), 78.7 (s), 75.8 (s), 75.4 (s),
73.1 (s), 71.6 (s), 39.8 (s), 31.9 (s), 29.33 (s), 29.27
(s), 29.21 (s), 27.0 (s), 22.7 (s), 14.2 (s); MALDI Calcd
for C70H88N2O11Na ([M+Na]+) 1155.6280; measure
1155.6236.
5.12. 2,20,3,30,4,40-Hexa-O-benzyl-a-D-trehalopyranuro-
nic acid (9)
To a solution of (COCl)2 (1.46mL, 17.0mmol) in 10mL
anhydrous CH2Cl2 at ꢁ78°C, anhydrous DMSO
(2.43mL, 34.2mmol) was added and the resulting solu-
tion was stirred for 15min, allowing the temperature
to be warmed up to ꢁ65°C. To the reaction flask, a
solution of 6 (5.0g, 5.7mmol) in 5mL CH2Cl2 was
added. The reaction mixture was stirred for 0.5h allow-
ing the temperature to be warmed up to ꢁ45°C. To this
solution, anhydrous Et3N (9.50mL, 68.4mmol) was
added, and the reaction was allowed to be warmed up
to ꢁ10–0°C in 0.5h. After completion of the reaction,
the reaction mixture quenched with 1N HCl and diluted
with Et2O. The combined organic layers were washed
with pH7 buffer (three times), brine, and dried over
Na2SO4. After removal of solvent, the crude product,
dialdehyde, (4.70g, 5.35mmol, 94%) was obtained as
viscous oil, and used without further purification. To
this solution of dialdehyde (0.5g, 0.57mmol) in 5mL
H2O–CH3CN (1:4) solution at 0°C, NaH2PO4 (0.03g,
0.6%), 30% H2O2 (0.14mL, 1.26mmol), and NaClO2
(0.16g, 17.1mmol) were added. The reaction mixture
was stirred for 12h, allowing the temperature to be
warmed up to room temperature. After the completion
of the reaction, the reaction was quenched with 0.1g
NaHSO3. The organic layer from the reaction was fil-
tered through a SPE column packed with silica gel.
The residue was washed with MeOH. After removal of
solvent, the product (0.41g, 0.45mmol, 79%) was af-
forded as pale yellowish viscous oil, and used without
further purification. 1H NMR (270MHz, CDCl3) d
7.10–7.50 (m, 30H), 5.17 (d, J = 3.3Hz, 2H, H-1), 5.03
(d, J = 10.9Hz, 2H, PhCH2O), 4.90 (d, J = 10.9Hz,
2H, PhCH2O), 4.83 (d, J = 10.6Hz, 2H, PhCH2O),
4.60–4.75 (m, 8H), 4.08 (t, J = 9.6Hz, 2H, H-3), 3.77
(t, J = 9.6Hz, 2H, H-4), 3.60 (dd, J = 9.6Hz,
J = 3.3Hz, 2H, H-2); 13C NMR (68MHz, CDCl3) d
174.6 (s, COOH), 138.5 (s), 137.5 (s), 137.5 (s), 128.6
(s), 128.4 (s), 128.2 (s), 126.0 (s), 127.8 (s), 95.0 (s, C-
1), 81.1 (s), 79.3 (s), 78.1 (s), 75.9 (s), 75.5 (s), 73.4 (s),
70.4 (s); LRFAB m/e 933 ([M+Na]+); HRFAB Calcd
for C54H54O13Na ([M+Na]+) m/e 933.3462; measure
m/e 933.3499.
5.14. Bis(N-octyl-a-D-glucopyranosyluronamide)ether
(YH012)
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 5.13 (d, J = 3.0Hz, 2H,
H-1), 4.26 (d, J = 9.9Hz, 2H, H-5), 3.79 (t, J = 9.9Hz,
2H, H-3), 3.55 (dd, J = 9.9Hz, J = 3.0Hz, 2H, H-2),
3.48 (t, J = 9.9Hz, 2H, H-4), 3.20 (t, J = 6.6Hz, 4H,
CONHCH2), 1.20–1.60 (m, 24H), 0.89 (t, J = 5.3Hz,
6H, CH2CH3); 13C NMR (68MHz, CD3OD) d 171.2
(s, CONH), 95.2 (s, C-1), 72.7 (s), 72.6 (s), 71.2 (s,
2C), 39.0 (s), 31.7 (s), 29.1 (s), 29.0 (s, 2 carbons), 26.6
(s), 22.4 (s), 13.1 (s); LRFAB m/e 615 ([M+Na]+);
HRFAB Calcd for C28H52N2O11Na ([M+Na]+) m/e
615.3469; measure m/e 615.3505.
5.15. Bis(2,3,4,-tri-O-benzyl-N-dodectyl-a-D-glucopyranos-
yluronamide)ether (11)
Please refer to the procedure for the preparation of 10.
1H NMR (270MHz, CDCl3) d 7.10–7.50 (m, 30H),
5.31 (t, J = 5.9Hz, 2H, CONH), 5.13 (d, J = 3.6Hz,
2H, H-1), 5.00 (d, J = 10.9Hz, 2H, PhCH2O), 4.92 (d,
J = 10.9Hz, 2H, PhCH2O), 4.78 (d, J = 12.2Hz, 2H,
PhCH2O), 4.78 (d, J = 10.2Hz, 2H, PhCH2O), 4.65 (d,
J = 12.2Hz, 2H, PhCH2O), 4.54 (d, J = 10.2Hz, 2H,
PhCH2O), 4.30 (d, J = 9.9Hz, 2H, H-5), 4.05 (t,
J = 9.2Hz, 2H, H-3), 3.68 (t, J = 9.2Hz, 2H, H-4),
3.61 (dd, J = 9.2Hz, J = 3.6Hz, 2H, H-2), 2.8–3.0 (m,
4H, CONHCH2), 1.00–2.00 (m, 40H), 0.87 (t,
J = 6.6Hz, 6H, CH2CH3); 13C NMR (68MHz, CDCl3)
d 168.5 (s, CONH), 138.5 (s), 138.2 (s), 138.0 (s), 128.7
(s), 128.5 (s), 128.2 (s), 128.0 (s), 127.8 (s), 127.7 (s),
127.0 (s), 94.8 (s, C-1), 81.4 (s), 80.3 (s), 78.7 (s), 75.8
(s), 75.4 (s), 73.0 (s), 71.6 (s), 39.8 (s), 32.0 (s), 29.8 (s,
2 carbons), 29.7 (s), 29.6 (s), 29.4 (s), 29.3 (s, 2 carbons),
27.0 (s), 22.8 (s), 14.2 (s); MALDI Calcd for
5.13. Bis(2,3,4,-tri-O-benzyl-N-octyl-a-D-glucopyranosyl-
uronamide)ether (10)
To a solution of 9 (0.1g, 0.11mmol) in anhydrous
CH2Cl2 (10mL) and DMF (0.02mL) at room tempera-
ture, (COCl)2 (0.024mL, 0.28mmol) was added. The
reaction was stirred for 1h. After removal of solvents,
the residue was redissolved in CH2Cl2 and added with

J. Wang et al. / Bioorg. Med. Chem. 12 (2004) 6397–6413
6405
C78H104N2O11Na ([M+Na]+) m/e 1267.7532; measure
m/e 1267.7590.
(s), 61.2 (s), 16.2 (s), 13.1 (s); LRFAB m/e 591
([M+Na]+); HRFAB Calcd for C22H36N2O15Na
([M+Na]+) m/e 591.2013; measure m/e 591.2002.
5.16. Bis(N-dodectyl-a-D-glucopyranosyluronamide)ether
(YH017)
5.19. Bis(methyl (6E)-2,3,4,-tri-O-benzyl-6,7-dideoxy-a-
D-gluco-oct-6-enopyranosyluronate)ether (14)
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 5.14 (d, J = 3.6Hz, 2H,
H-1), 4.27 (d, J = 9.9Hz, 2H, H-5), 2.80–3.80 (m, 10H),
1.10–1.80 (m, 40H), 0.92 (t, J = 6.9Hz, 6H, CH2CH3);
13C NMR (68MHz, CD3OD) d 168.0 (s, CONH), 95.0
(s, C-1), 72.7 (s), 72.5 (s), 71.2 (s, 2 carbons), 38.9 (s),
31.7 (s), 29.42 (s, 2 carbons), 29.40 (s, 2 carbons), 29.1
(s, 2 carbons), 29.0 (s), 26.6 (s), 22.4 (s), 13.1 (s); LRFAB
m/e 727 ([M+Na]+); HRFAB Calcd for C36H68N2O11Na
([M+Na]+) m/e 727.4721; measure m/e 727.4720.
The dialdehyde was prepared according to the proce-
dure described in the preparation of 9. To a solution
of methyl diethylphosphonoacetate (0.38g, 1.8mmol)
in anhydrous THF at 0°C, NaH (0.044g, 1.8mmol)
was added. After being stirred for 1h, a solution of dial-
dehyde (0.50g, 0.57mmol) was added. The reaction was
stirred for another 12h. After completion of the reac-
tion, saturated NH4Cl(aq) and Et2O were added. The
combined organic solution was washed with brine and
dried over Na2SO4(s). After removal of the solvents,
followed by purification with a gradient column
chromatography (hexane–EtOAc = 100:0 to 60:40), the
product was obtained as a pale yellowish oil (0.46g,
5.17. Bis(2,3,4,-tri-O-benzyl-N-((R)-1-ethoxycarbonyl-
ethyl)-a-D-glucopyranosyluronamide)ether (12)
1
To a solution of 9 (1.0g, 1.10mmol) in pyridine, ethyl L-
alanine ester HCl (0.55g, 4.0mmol) and EDCÆHCl
(0.76g, 4.0mmol) were added. The reaction was stirred
for 12h at 50°C. After completion of the reaction, the
reaction mixture was diluted with EtOAc. The combined
organic solution was washed with 1N HCl(aq), water,
saturated NaHCO3(aq), and brine, then dried over
Na2SO4(s). After removal of the solvent, followed by
purification with a gradient column chromatography
(hexane–EtOAc = 75:25 to 25:75), the product was ob-
tained as a yellowish viscous oil (0.3g, 0.27mmol, 25%).
1H NMR (270MHz, CDCl3) d 7.10–7.50 (m, 30H), 6.53
(d, J = 6.9Hz, 2H, CONH), 5.10 (d, J = 3.6Hz, 2H,
H-1), 4.99 (d, J = 10.9Hz, 2H, PhCH2O), 4.89 (d,
J = 10.9Hz, 2H, PhCH2O), 4.80 (d, J = 10.2Hz, 2H,
PhCH2O), 4.70 (d, J = 10.2Hz, 2H, PhCH2O), 4.68 (s,
4H, PhCH2O), 4.54 (d, J = 9.9Hz, 2H, H-5), 4.42 (dt,
J = 6.9Hz, J = 7.2Hz, 2H, COCH(CH3)NH), 4.13 (q,
J = 6.9Hz, 4H, CH3CH2OOC), 4.05 (t, J = 9.9Hz, 2H,
H-3), 3.66 (t, J = 9.9Hz, 2H, H-4), 3.56 (dd, J = 9.9Hz,
J = 3.6Hz, 2H, H-2), 1.23 (d, J = 7.2Hz, 6H,
COCH(CH3)NH), 1.18 (t, J = 6.9Hz, 6H, CH3CH2-
OOC); 13C NMR (68MHz, CDCl3) d 172.5 (s,
COOCH2CH3), 168.5 (s, CONH), 138.6 (s), 138.0 (s),
137.9 (s), 128.6 (s), 128.4 (s), 128.0 (s), 127.9 (s), 127.8
(s), 127.7 (s), 95.4 (s, C-1), 81.4 (s), 80.2 (s), 78.1 (s),
75.8 (s), 75.4 (s), 73.0 (s), 71.1 (s), 61.5 (s), 48.1 (s), 18.1
(s), 14.2 (s); MALDI Calcd for C64H72N2O15Na
([M+Na]+) m/e 1131.4825; measure m/e 1131.4779.
0.46mmol, 82%). H NMR (270MHz, CDCl3) d 7.1–
7.5 (m, 30H), 6.95 (dd, J = 15.5Hz, J = 4.3Hz, 2H,
COCH@CH), 6.00 (dd, J = 15.5Hz, J = 2.0Hz, 2H,
COCH@CH), 5.09 (d, J = 3.6Hz, 2H, H-1), 5.00 (d,
J = 10.9Hz, 2H, PhCH2O), 4.87 (d, J = 10.9Hz, 2H,
PhCH2O), 4.78 (d, J = 10.6Hz, 2H, PhCH2O), 4.40–
4.77 (m, 8H), 4.03 (t, J = 9.6Hz, 2H, H-3), 3.74 (s,
6H, COOCH3), 3.50 (dd, J = 9.6Hz, J = 3.6Hz, 2H,
H-2), 3.21 (t, J = 9.6Hz, 2H, H-4); 13C NMR
(68MHz, CDCl3) d 166.7 (s, COCH@CH), 144.6 (s,
COCH@CH), 138.7 (s), 137.8 (s), 137.6 (s), 128.6 (s),
128.5 (s), 128.4 (s), 128.3 (s), 127.9 (s), 127.8 (s), 127.7
(s), 121.1 (s, COCH@CH), 94.3 (s, C-1), 82.0 (s), 81.6
(s), 79.1 (s), 75.9 (s), 75.6 (s), 73.2 (s), 69.9 (s), 51.7 (s);
MALDI Calcd for C60H62O13Na ([M+Na]+) m/e
1013.4083; measure m/e 1013.4150.
General procedure for transamidation: The procedure
was modified from the reported procedure of Ref. 14.
To a solution of alkylamine in freshly distilled anhy-
drous CH2Cl2 (1mL), Al(Me)3 (1equiv, from a 2.0M
solution in toluene) was slowly added. After being stir-
red for 0.5h, a solution of 14 (0.17equiv) dissolved in
freshly distilled anhydrous CH2Cl2 (1mL) was added.
The mixture was stirred overnight at room temperature.
After completion of the reaction (analyzed by TLC,
eluted with hexane–EtOAc (1:3) solution), the reaction
was quenched by the slow addition of 1N HCl, then di-
luted with EtOAc. The combined organic solution was
washed with 1N HCl, H2O, NaHCO3, and brine then
dried over Na2SO4(s). After removal of the solvents,
followed by purification with a gradient column
chromatography (hexane–EtOAc = 90:10 to 20:80), the
product was obtained as a pale yellowish oil.
5.18. Bis(N-((R)-1-ethoxycarbonylethyl)-a-D-glucopyranos-
yluronamide)ether (YH024)
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 5.17 (d, J = 3.3Hz, 2H,
H-1), 4.45 (q, J = 7.2Hz, 2H, CONHCH(CH3)), 4.35 (d,
J = 9.9Hz, 2H, H-5), 4.20 (q, J = 6.9Hz, 4H,
CH3CH2OOC), 3.80 (t, J = 9.9Hz, 2H, H-3), 3.52 (dd,
J = 9.9Hz, J = 3.3Hz, 2H, H-2), 3.49 (t, J = 9.9Hz,
2H, H-4), 1.23 (d, J = 7.2Hz, 6H, COCH(CH3)NH),
1.18 (t, J = 6.9Hz, 6H, CH3CH2OOC); 13C NMR
(68MHz, CD3OD) d 172.7 (s, COOCH2CH3), 170.9 (s,
CONH), 95.1 (s, C-1), 72.6 (s), 72.5 (s), 71.2 (s), 71.1
5.20. Bis((6E)-2,3,4,-tri-O-benzyl-6,7-dideoxy-N-octyl-a-
D-gluco-oct-6-enopyranuronamide)ether (15)
Please refer to the general procedure for transamidation.
1H NMR (270MHz, CDCl3) d 7.3–7.4 (m, 30H), 6.78
(dd, J = 5.9Hz, J = 15.2Hz, 2H), 5.56 (d, J = 15.2Hz,
2H), 5.16 (d, J = 3.6Hz, 2H, H-1), 5.11 (t, J = 5.9Hz,
2H, C(O)NH), 5.01 (d, J = 10.9Hz, 2H, PhCH2O),

6406
J. Wang et al. / Bioorg. Med. Chem. 12 (2004) 6397–6413
4.92 (d, J = 10.9Hz, 2H, PhCH2O), 4.78 (d, J = 10.6Hz,
2H, PhCH2O), 4.6–4.7 (m, 6H), 4.52 (d, J = 10.6Hz, 2H,
PhCH2O), 4.07 (dd, J = 9.2Hz, J = 9.2Hz, 2H), 3.57
(dd, J = 3.6Hz, J = 9.6Hz, 2H, H-2), 3.27 (m, 6H),
1.2–1.5 (m, 24H), 0.87 (t, J = 6.4Hz, 6H); 13C NMR
(68MHz, CDCl3) d 165.0 (s), 139.4 (s), 138.7 (s), 138.5
(s), 137.9 (s), 128.5 (s), 128.4 (s), 128.0 (s), 127.8 (s),
127.44 (s), 127.38 (s), 125.3 (s), 93.9 (s, C-1), 82.2 (s),
81.5 (s), 79.6 (s), 75.9 (s), 75.6 (s), 73.0 (s), 70.7 (s),
39.6 (s), 31.9 (s), 29.8 (s), 29.4 (s), 29.3 (s), 27.1 (s),
22.8 (s), 14.2 (s); MALDI Calcd for C74H92N2O11Na
([M+Na]+) m/e 1207.6593; measure m/e 1207.6593.
J = 3.0Hz, 2H, H-1), 5.01(d, J = 10.9Hz, 2H, PhCH2O),
4.92 (d, J = 10.9Hz, 2H, PhCH2O), 4.78 (d, J = 12.2Hz,
2H, PhCH2O), 4.6–4.7 (m, 6H), 4.52 (d, J = 10.5Hz, 2H,
PhCH2O), 4.08 (m, 2H), 3.56 (dd, J = 9.6Hz, J = 5.9Hz,
2H, H-2), 3.2–3.3 (m, 6H), 1.5 (m, 4H), 1.2–1.3 (m, 8H),
0.91 (t, J = 5.3Hz, 6H); 13C NMR (68MHz, CDCl3) d
165.0 (s), 139.4 (s), 138.7 (s), 138.4 (s), 137.9 (s), 128.5
(s), 128.4 (s), 128.0 (s), 127.7 (s), 127.4 (s), 125.3 (s),
93.92 (s, C-1), 82.2 (s), 81.5 (s), 79.6 (s), 75.9 (s), 75.6
(s), 73.0 (s), 70.6 (s), 39.6 (s), 29.4 (s), 29.2 (s), 22.5 (s),
14.1 (s); MALDI Calcd for C68H80N2O11Na
([M+Na]+) m/e 1123.5654; measure m/e 1123.5667.
5.21. Bis((6E)-2,3,4,-tri-O-benzyl-6,7-dideoxy-N-dodect-
yl-a-D-gluco-oct-6-enopyranuro-namide)ether (16)
5.24. Bis((6E)-2,3,4,-tri-O-benzyl-6,7-dideoxy-N-hexyl-a-
D-gluco-oct-6-enopyranuronamide)ether (19)
Please refer to the general procedure for transamidation.
1H NMR (270MHz, CDCl3) d 7.2–7.3 (m, 30H), 6.77
(dd, J = 5.9Hz, J = 15.2Hz, 2H), 5.55 (d, J = 15.2Hz,
2H), 5.14 (d, J = 3.3Hz, 2H, H-1), 5.12 (t, 2H,
C(O)NH), 5.00 (d, J = 10.9Hz, 2H, PhCH2O), 4.91 (d,
J = 10.9Hz, 2H, PhCH2O), 4.77 (d, J = 10.6Hz, 2H,
PhCH2O), 4.6–4.7 (m, 6H), 4.51 (d, J = 10.2Hz, 2H,
PhCH2O), 4.07 (dd, J = 9.2Hz, J = 9.2Hz, 2H), 3.56
(dd, J = 3.3Hz, J = 9.6Hz, 2H, H-2), 3.2–3.3 (m, 6H),
1.2–1.5 (m, 40H), 0.87 (t, 6H); 13C NMR (68MHz,
CDCl3) d 165.0 (s), 139.3 (s), 138.7 (s), 138.4 (s), 137.9
(s), 128.5 (s), 128.4 (s), 128.0 (s), 127.7 (s), 127.4 (s),
125.3 (s), 93.9 (s, C-1), 82.2 (s), 81.5 (s), 79.6 (s), 75.9
(s), 75.6 (s), 72.9 (s), 70.6 (s), 39.6 (s), 32.0 (s, 2 carbons),
31.0 (s), 29.7 (s, 3 carbons), 29.4 (s, 2 carbons), 27.1 (s),
22.8 (s), 14.2 (s). LRFAB C82H108N2O11K ([M+K]+)
m/e 1336.
Please refer to the general procedure for transamidation.
1H NMR (270MHz, CDCl3) d 7.2–7.4 (m, 30H), 6.78
(dd, J = 5.9Hz, J = 15.2Hz, 2H), 5.56 (d, J = 15.2Hz,
2H), 5.15 (d, J = 3.3Hz, 2H, H-1), 5.13 (t, J = 5.9Hz,
2H, C(O)NH), 5.00 (d, J = 10.9Hz, 2H, PhCH2O), 4.91
(d, J = 10.9Hz, 2H, PhCH2O), 4.77 (d, J = 10.6Hz,
2H, PhCH2O), 4.6–4.7 (m, 6H), 4.51 (d, J = 10.5Hz,
2H, PhCH2O), 4.07 (dd, J = 9.2Hz, J = 9.2Hz, 2H),
3.55 (dd, J = 3.3Hz, J = 9.6Hz, 2H, H-2), 3.2–3.3 (m,
6H), 1.4–1.5 (m, 4H), 1.2–1.3 (m, 12H), 0.87 (t,
J = 6.6Hz, 6H); 13C NMR (68MHz, CDCl3) d 165.0
(s), 139.4 (s), 138.7 (s), 138.4 (s), 137.9 (s), 128.5 (s),
128.4 (s), 128.0 (s), 127.7 (s), 127.4 (s), 125.3 (s), 93.9
(s, C-1), 82.2 (s), 81.5 (s), 79.6 (s), 75.9 (s), 75.6 (s),
72.9 (s), 70.6 (s), 39.6 (s), 31.6 (s), 29.7 (s), 26.0 (s), 22.7
(s), 14.1 (s); MALDI Calcd for C70H84N2O11Na
([M+Na]+) m/e 1151.5967; measure m/e 1151.5970.
5.22. Bis((6E)-2,3,4,-tri-O-benzyl-6,7-dideoxy-N-butyl-a-
D-gluco-oct-6-enopyranuronamide)ether (17)
5.25. Bis((6E)-2,3,4,-tri-O-benzyl-6,7-dideoxy-N-heptyl-
a-D-gluco-oct-6-enopyranuronamide)ether (20)
Please refer to the general procedure for transamidation.
1H NMR (270MHz, CDCl3) d 7.2–7.4 (m, 30H), 6.81
(dd, J = 5.9Hz, J = 15.2Hz, 2H), 5.61 (d, J = 15.2Hz,
2H), 5.27 (t, J = 5.9Hz, 2H, C(O)NH), 5.16 (d,
J = 3.3Hz, 2H, H-1), 5.01 (d, J = 10.9Hz, 2H,
PhCH2O), 4.92 (d, J = 10.9Hz, 2H, PhCH2O), 4.78 (d,
J = 10.5Hz, 2H, PhCH2O), 4.73 (d, J = 12.2Hz, 2H,
PhCH2O), 4.6 (m, 2H), 4.64 (d, J = 12.2Hz, 2H,
PhCH2O), 4.53 (d, J = 10.5Hz, 2H, PhCH2O), 4.09
(dd, J = 9.6Hz, J = 9.6Hz, 2H), 3.58 (dd, J = 3.3Hz,
J = 9.9Hz, 2H, H-2), 3.2–3.3 (m, 6H), 1.2–1.5 (m, 8H),
0.93 (t, J = 7.1Hz, 6H); 13C NMR (68MHz, CDCl3) d
165.1 (s), 139.3 (s), 138.7 (s), 138.4 (s), 138.0 (s), 128.5
(s), 128.4 (s), 128.0 (s), 127.7 (s), 127.5 (s), 127.4 (s),
125.3 (s), 93.9 (s, C-1), 82.2 (s), 81.5 (s), 79.6 (s), 75.9
(s), 75.6 (s), 73.0 (s), 70.6 (s), 39.3 (s), 31.8 (s), 20.2 (s),
13.9 (s); MALDI Calcd for C66H76N2O11Na
([M+Na]+) m/e 1095.5341; measure m/e 1095.5322.
Please refer to the general procedure for transamidation.
1H NMR (270MHz, CDCl3) d 7.1–7.5 (m, 30H), 6.80
(dd, J = 5.9Hz, J = 15.2Hz, 2H), 5.58 (d, J = 15.2Hz,
2H), 5.16 (d, J = 3.3Hz, 2H, H-1), 5.01 (d,
J = 10.9Hz, 2H, PhCH2O), 4.92 (d, J = 10.9Hz, 2H,
PhCH2O), 4.79 (d, J = 10.6Hz, 2H, PhCH2O), 4.6–4.7
(m, 6H), 4.53 (d, J = 10.6Hz, 2H, PhCH2O), 4.09 (dd,
J = 9.2Hz, J = 9.2Hz, 2H), 3.57 (dd, J = 3.6Hz,
J = 9.6Hz, 2H, H-2), 3.2–3.3 (m, 6H), 1.5–1.6 (m, 4H),
1.2–1.4 (m, 16H), 0.90 (t, J = 6.9Hz, 6H); 13C NMR
(68MHz, CDCl3) d 165.0 (s), 139.4 (s), 138.7 (s), 138.5
(s), 138.0 (s), 128.5 (s), 128.4 (s), 128.0 (s), 127.8 (s),
127.4 (s), 125.3 (s), 93.9 (s, C-1), 82.2 (s), 81.5 (s), 79.6
(s), 75.9 (s), 75.6 (s), 73.0 (s), 70.6 (s), 39.6 (s), 31.9 (s),
29.8 (s), 29.1 (s), 27.0 (s), 22.7 (s), 14.2 (s); MALDI
Calcd for C72H88N2O11Na ([M+Na]+) m/e 1179.6280;
measure m/e 1179.6240.
5.26. Bis((6E)-2,3,4,-tri-O-benzyl-6,7-dideoxy-N-nonyl-a-
D-gluco-oct-6-enopyranuronamide)ether (21)
5.23. Bis((6E)-2,3,4,-tri-O-benzyl-6,7-dideoxy-N-pentyl-
a-D-gluco-oct-6-enopyranuronamide)ether (18)
Please refer to the general procedure for transamidation.
1H NMR (270MHz, CDCl3) d 7.3–7.4 (m, 30H), 6.80
(dd, J = 5.9Hz, J = 15.2Hz, 2H), 5.58 (d, J = 15.2Hz,
2H), 5.16 (d, J = 3.3Hz, 2H, H-1), 5.15 (m, 2H,
C(O)NH), 5.00 (d, J = 10.9Hz, 2H, PhCH2O), 4.92 (d,
Please refer to the general procedure for transamidation.
1H NMR (270MHz, CDCl3) d 7.2–7.4 (m, 30H), 6.79
(dd, J = 5.9Hz, J = 15.2Hz, 2H), 5.59 (d, J = 15.2Hz,
2H), 5.18 (t, J = 5.9Hz, 2H, C(O)NH), 5.16 (d,

J. Wang et al. / Bioorg. Med. Chem. 12 (2004) 6397–6413
6407
J = 10.9Hz, 2H, PhCH2O), 4.78 (d, J = 10.9Hz, 2H,
PhCH2O), 4.6–4.7 (m, 6H), 4.52 (d, J = 10.9Hz, 2H,
PhCH2O), 4.09 (dd, J = 9.2Hz, J = 9.2Hz, 2H), 3.57
(dd, J = 3.3Hz, J = 9.6Hz, 2H, H-2), 3.2–3.3 (m, 6H),
1.2–1.5 (m, 28H), 0.89 (t, J = 7.3Hz, 6H); 13C NMR
(68MHz, CDCl3) d 165.0 (s), 139.4 (s), 138.7 (s), 138.4
(s), 137.9 (s), 128.5 (s), 128.4 (s), 128.0 (s), 127.8 (s),
127.44 (s), 127.40 (s), 125.3 (s), 93.93 (s, C-1), 82.2 (s),
81.5 (s), 79.6 (s), 75.9 (s), 75.6 (s), 73.0 (s), 70.6 (s), 39.6
(s), 32.0 (s), 29.8 (s), 29.6 (s), 29.5 (s), 29.4 (s), 27.1 (s),
22.8 (s), 14.2 (s); MALDI Calcd for C76H96N2O11Na
([M+Na]+) m/e 1235.6906; measure m/e 1235.6970.
J = 10.9Hz, 2H, PhCH2O), 4.5–4.8 (m, 10H), 4.08 (m,
2H), 3.2–3.6 (m, 16H), 1.2–1.4 (m, 12H), 0.90 (t,
J = 7.3Hz, 6H); 13C NMR (68MHz, CDCl3) d 165.1
(s, C(O)NH), 139.5 (s), 138.7 (s), 138.2 (s), 137.9 (s),
128.5 (s), 128.4 (s), 128.0 (s), 127.7 (s), 127.6 (s), 124.9
(s), 94.1 (s, C-1), 82.3 (s), 81.5 (s), 79.4 (s), 75.9 (s),
75.6 (s), 73.1 (s), 71.1 (s), 70.5 (s), 69.6 (s), 38.2 (s),
31.9 (s), 29.3 (s), 19.5 (s), 14.1 (s); MALDI Calcd for
C72H88N2O13Na ([M+Na]+) m/e 1211.6179; measure
m/e 1211.6192.
5.30. Bis(N-octyl-a-D-gluco-octopyranuronamide)ether
(YH023)
5.27. Bis((6E)-2,3,4,-tri-O-benzyl-6,7-dideoxy-N-decyl-a-
D-gluco-oct-6-enopyranuronamide)ether (22)
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 5.00 (d, J = 3.0Hz, 2H,
H-1), 3.7–3.9 (m, 4H), 3.46 (dd, J = 9.6Hz, J = 3.0Hz,
2H, H-2), 3.0–3.2 (m, 4H), 3.09 (dd, J = 8.9Hz,
J = 9.2Hz, 4H), 2.4–2.6 (m, 8H), 1.3–2.0 (m, 24H),
0.89 (t, J = 7.6Hz, 6H, CH2CH3); 13C NMR (68MHz,
CD3OD) d 174.8 (s, CONH), 94.0 (s, C-1), 74.4 (s),
73.2 (s), 72.1 (s), 71.0 (s), 39.2 (s), 32.1 (s), 31.7 (s),
31.6 (s), 29.1 (s), 28.9 (s), 27.8 (s), 26.8 (s), 22.4 (s),
13.2 (s); LRFAB m/e 671 ([M+Na]+); HRFAB Calcd
for C32H60N2O11Na ([M+Na]+) m/e 671.4095; measure
m/e 671.4121.
Please refer to the general procedure for transamidation.
1H NMR (270MHz, CDCl3) d 7.2–7.3 (m, 30H), 6.79
(dd, J = 5.9Hz, J = 15.2Hz, 2H), 5.57 (d, J = 15.2Hz,
2H), 5.15 (d, J = 3.3Hz, 2H, H-1), 5.14 (m, 2H,
C(O)NH), 5.01 (d, J = 10.6Hz, 2H, PhCH2O), 4.92 (d,
J = 10.6Hz, 2H, PhCH2O), 4.78 (d, J = 10.6Hz, 2H,
PhCH2O), 4.6–4.7 (m, 6H), 4.52 (d, J = 10.6Hz, 2H,
PhCH2O), 4.07 (dd, J = 9.2Hz, J = 9.2Hz, 2H), 3.57
(dd, J = 3.3Hz, J = 9.6Hz, 2H, H-2), 3.2–3.3 (m, 6H),
1.2–1.5 (m, 32H), 0.89 (t, J = 6.4Hz, 6H); 13C NMR
(68MHz, CDCl3) d 165.0 (s), 139.4 (s), 138.7 (s), 138.4
(s), 137.9 (s), 128.5 (s), 128.4 (s), 128.0 (s), 127.7 (s),
127.4 (s), 125.3 (s), 93.9 (s, C-1), 82.2 (s), 81.5 (s), 79.6
(s), 75.9 (s), 75.6 (s), 73.0 (s), 70.6 (s), 39.6 (s), 32.0 (s),
29.8 (s), 29.7 (s, 2 carbons), 29.44 (s), 29.42 (s), 27.1
(s), 22.8 (s), 14.2 (s); MALDI Calcd for C78H100N2O11-
Na ([M+Na]+) m/e 1263.7219; measure m/e 1263.7253.
5.31. Bis(N-dodectyl-a-D-gluco-octopyranuronamide)-
ether (YH027)
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 5.00 (d, J = 3.6Hz, 2H,
H-1), 3.7–3.8 (m, 4H), 3.46 (dd, J = 9.6Hz, J = 3.6Hz,
2H), 3.0–3.2 (m, 6H), 2.2–2.4 (m, 4H), 2.1–2.2 (m,
4H), 1.6–1.7 (m, 4H), 1.4–1.5 (m, 4H), 1.28–1.30 (m,
32H), 0.86–0.91 (m, 6H); 13C NMR (68MHz, CD3OD)
d 174.8 (s), 93.9 (s, C-1), 74.4 (s), 73.2 (s), 72.1 (s), 71.0
(s), 39.1 (s), 32.1 (s), 31.8 (s), 29.4 (s, 3 carbons), 29.2 (s,
3 carbons), 29.1 (s), 27.7 (s), 26.8 (s), 22.4 (s), 13.1 (s);
LRFAB m/e 784 ([M+Na]+); HRFAB Calcd for
C40H76N2O11Na ([M+Na]+) m/e 783.5347; measure
m/e 783.5364.
5.28. Bis((6E)-2,3,4,-tri-O-benzyl-6,7-dideoxy-N-(3-eth-
oxypropyl)-a-D-gluco-oct-6-enopyranuronamide)ether (23)
Please refer to the general procedure for transamidation.
1H NMR (270MHz, CDCl3) d 7.2–7.3 (m, 30H), 6.83
(dd, J = 5.6Hz, J = 15.2Hz, 2H), 5.79 (t, J = 4.9Hz,
2H, C(O)NH), 5.69 (d, J = 15.2Hz, 2H), 5.15 (d,
J = 3.3Hz, 2H, H-1), 5.00 (d, J = 10.9Hz, 2H,
PhCH2O), 4.90 (d, J = 10.9Hz, 2H, PhCH2O), 4.77 (d,
J = 10.6Hz, 2H, PhCH2O), 4.6–4.7 (m, 6H), 4.53 (d,
J = 10.6Hz, 2H, PhCH2O), 4.07 (dd, J = 9.6Hz,
J = 9.6Hz, 2H), 3.54 (dd, J = 3.3Hz, J = 9.6Hz, 2H,
H-2), 3.4–3.5 (m, 12H), 3.24 (dd, J = 9.6Hz,
J = 9.6Hz, 2H), 1.79 (m, 4H), 1.17 (t, J = 7.1Hz, 6H);
13C NMR (68MHz, CDCl3) d 165.1 (s, C(O)NH),
139.5 (s), 138.7 (s), 138.2 (s), 137.9 (s), 128.5 (s), 128.4
(s), 128.0 (s), 127.7 (s), 127.6 (s), 125.0 (s), 94.0 (s, C-
1), 82.3 (s), 81.5 (s), 79.4 (s), 75.9 (s), 75.6 (s), 73.0 (s),
70.5 (s), 69.3 (s), 66.5 (s), 38.1 (s), 29.4 (s), 15.3 (s);
MALDI Calcd for C68H80N2O13Na ([M+Na]+) m/e
1155.5553; measure m/e 1155.5618.
5.32. Bis(N-butyl-a-D-gluco-octopyranuronamide)ether
(YH028)
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 4.99 (d, J = 3.6Hz, 2H,
H-1), 3.7–3.8 (m, 4H), 3.45 (m, 2H), 3.31 (d, J = 10.8Hz,
2H), 3.17 (t, J = 6.3Hz, 2H), 3.07 (t, J = 9.3Hz, 2H),
2.2–2.4 (m, 4H), 2.1–2.2 (m, 4H), 1.3–1.7 (m, 8H), 0.92
(t, J = 8.1Hz, 6H); 13C NMR (68MHz, CD3OD) d
175.2 (s), 94.1 (s, C-1), 74.4 (s), 73.2 (s), 72.1 (s), 70.9
(s), 39.2 (s), 31.9 (s), 31.0 (s), 27.8 (s), 19.8 (s), 12.8 (s);
LRFAB m/e 559 ([M+Na]+); HRFAB Calcd for
C24H44N2O11Na ([M+Na]+) m/e 559.2843; measure
m/e 559.2845.
5.29. Bis((6E)-2,3,4,-tri-O-benzyl-6,7-dideoxy-N-(3-but-
oxypropyl)-a-D-gluco-oct-6-enopyranuronamide)ether (24)
5.33. Bis(N-pentyl-a-D-gluco-octopyranuronamide)ether
(YH029)
Please refer to the general procedure for transamidation.
1H NMR (270MHz, CDCl3) d 7.3–7.4 (m, 30H), 6.86
(m, 2H), 5.7–5.8 (m, 4H), 5.12 (d, J = 3.9Hz, 2H, H-
1), 4.99 (d, J = 10.9Hz, 2H, PhCH2O), 4.92 (d,
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 4.96 (d, 2H, H-1

6408
J. Wang et al. / Bioorg. Med. Chem. 12 (2004) 6397–6413
protons obscured by water peak), 3.7–3.8 (m, 4H), 3.47
(d, J = 6.9Hz, 2H), 3.2–3.3 (m, 4H), 3.08 (t, J = 9.2Hz,
2H), 2.4–2.5 (m, 4H), 2.2 (m, 4H), 1.5 (m, 4H), 1.3 (m,
8H), 0.9 (m, 6H); 13C NMR (68MHz, CD3OD) d
175.6 (s), 94.3 (s, C-1), 74.3 (s), 73.1 (s), 72.1 (s), 70.9
(s), 39.8 (s), 31.7 (s), 28.9 (s), 28.5 (s), 27.8 (s), 22.1 (s),
13.1 (s); LRFAB m/e 587 ([M+Na]+); HRFAB Calcd
for C26H48N2O11Na ([M+Na]+) m/e 587.3156; measure
m/e 587.3155.
(s), 27.8 (s), 26.8 (s), 22.4 (s), 13.2 (s); MALDI Calcd
for C36H68N2O11Na ([M+Na]+) m/e 727.4715; measure
m/e 727.4726.
5.38. Bis(N-(3-ethoxypropyl)-a-D-gluco-octopyranuron-
amide)ether (YH034)
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 4.99 (d, J = 3.9Hz, 2H,
H-1), 3.7–3.8 (m, 4H), 3.4–3.5 (m, 8H), 3.2–3.3 (m, 6H),
3.06 (dd, J = 8.9Hz, J = 8.9Hz, 2H), 2.1–2.4 (m, 8H),
1.7–1.8 (m, 4H), 1.16 (t, J = 6.8Hz, 6H); 13C NMR
(68MHz, CD3OD) d 175.0 (s), 94.0 (s, C-1), 74.4 (s),
73.2 (s), 72.0 (s), 70.9 (s), 67.9 (s), 66.0 (s), 36.6 (s),
32.0 (s), 29.2 (s), 27.7 (s), 14.2 (s); MALDI Calcd for
C26H48N2O13Na ([M+Na]+) m/e 619.3049; measure
m/e 619.3064.
5.34. Bis(N-hexyl-a-D-gluco-octopyranuronamide)ether
(YH030)
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 4.99 (d, J = 3.6Hz, 2H,
H-1), 3.7–3.8 (m, 4H), 3.46 (dd, J = 9.6Hz, J = 3.6Hz,
2H), 3.3–3.4 (m, 2H), 3.13 (t, J = 6.9Hz, 2H), 3.06
(dd, J = 9.2Hz, J = 9.2Hz, 2H), 2.1–2.4 (m, 8H), 1.6–
1.7 (m, 4H), 1.4–1.5 (m, 4H), 1.2–1.3 (m, 8H), 0.9 (m,
6H); 13C NMR (68MHz, CD3OD) d 174.9 (s), 94.0 (s,
C-1), 74.4 (s), 73.2 (s), 72.1 (s), 70.9 (s), 39.2 (s), 32.1
(s), 31.4 (s), 29.0 (s), 27.8 (s), 26.4 (s), 22.3 (s), 13.1 (s);
LRFAB m/e 615 ([M+Na]+); HRFAB Calcd for
C28H52N2O11Na ([M+Na]+) m/e 615.3469; measure
m/e 615.3473.
5.39. Bis(N-(3-butoxypropyl)-a-D-gluco-octopyranuron-
amide)ether (YH035)
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 5.00 (d, J = 4.0Hz, 2H,
H-1), 3.7–3.8 (m, 4H), 3.4–3.5 (m, 8H), 3.29–3.34 (m,
4H), 3.23 (t, J = 6.9Hz, 2H), 3.07 (dd, J = 9.6Hz,
J = 8.9Hz, 2H), 2.1–2.4 (m, 8H), 1.7–1.8 (m, 4H), 1.5–
1.6 (m, 4H), 1.3–1.4 (m, 4H), 0.92 (t, J = 7.3Hz, 6H);
13C NMR (68MHz, CD3OD) d 174.79 (s, C(O)NH),
94.0 (s, C-1), 74.5 (s), 73.2 (s), 72.1 (s), 70.9 (s), 70.5
(s), 68.2 (s), 36.6 (s), 32.1 (s), 31.6 (s), 29.2 (s), 27.8 (s),
19.1 (s), 13.0 (s); MALDI Calcd for C30H56N2O13Na
([M+Na]+) m/e 675.3675; measure m/e 675.3702.
5.35. Bis(N-heptyl-a-D-gluco-octopyranuronamide)ether
(YH031)
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 4.98 (s, 2H, H-1), 3.7–
3.8 (m, 4H), 3.46 (m, 2H), 3.29 (s, 2H), 3.0–3.2 (m, 4H),
2.1–2.4 (m, 8H), 1.5–1.7 (m, 8H), 1.3 (m, 12H), 0.9 (m,
6H); 13C NMR (68MHz, CD3OD) d 175.3 (s), 94.2 (s,
C-1), 74.4 (s), 73.2 (s), 72.1 (s), 70.9 (s), 39.6 (s), 31.9
(s), 31.6 (s), 28.9 (s), 28.8 (s), 27.8 (s), 26.7 (s), 22.4 (s),
13.1 (s); MALDI Calcd for C30H56N2O11Na
([M+Na]+) m/e 643.3776; measure m/e 643.3758.
5.40. 6,60-Diazido-2,20,3,30,4,40-hexa-O-benzyl-6,60-dide-
oxy-a-D-trehalose (25)
To a solution of 6 (1.0g, 1.1mmol) in anhydrous
CH2Cl2 (10mL) and pyridine (0.26mL, 3.2mmol) at
0°C, Tf2O (0.46mL, 2.7mmol) was slowly added. After
completion of the reaction (ca. 30min), the reaction mix-
ture was diluted with CH2Cl2. The combined organic
5.36. Bis(N-nonyl-a-D-gluco-octopyranuronamide)ether
(YH032)
layers were washed with water, saturated NaHCO3(aq)
,
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 4.93 (d, J = 3.0Hz, 2H,
H-1), 3.6–3.8 (m, 4H), 3.46 (m, 2H), 3.1–3.3 (m, 6H),
2.1–2.5 (m, 8H), 1.2–1.7 (m, 28H), 0.9 (m, 6H); 13C
NMR (68MHz, CD3OD) d 175.6 (s), 94.3 (s, C-1),
74.4 (s), 73.2 (s), 72.1 (s), 70.9 (s), 39.9 (s), 39.8 (s),
31.7 (s), 29.4 (s), 29.1 (s, 2 carbons), 28.8 (s), 27.8 (s),
26.7 (s), 22.4 (s), 13.2 (s); MALDI Calcd for
C34H64N2O11Na ([M+Na]+) m/e 699.4402; measure
m/e 699.4372.
and brine, then dried over Na2SO4(s). After removal of
solvent, the triflated crude product was dissolved in
DMF (10mL), and added with excess NaN3. The reac-
tion was stirred for 12h. After removal of DMF, EtOAc
was added, and the resulting cloudy yellowish solution
was filtered through Celite. After removal of solvents
followed by a gradient column chromatography (hex-
ane–EtOAc = 90:10 to 65:35), the product was afforded
as a pale yellowish oil (0.78g, 0.84mmol, 74%). 1H
NMR (270MHz, CDCl3) d 7.10–7.50 (m, 30H), 5.26
(d, J = 3.6Hz, 2H, H-1), 5.03 (d, J = 10.8Hz, 2H,
PhCH2O), 4.93 (d, J = 10.0Hz, 2H, PhCH2O), 4.90 (d,
J = 10.8Hz, 2H, PhCH2O), 4.77 (d, J = 12.0Hz, 2H,
PhCH2O), 4.74 (d, J = 12.0Hz, 2H, PhCH2O), 4.62 (d,
J = 10.0Hz, 2H, PhCH2O), 4.21 (dt, J = 9.6Hz,
J = 2.8Hz, 2H, H-5), 4.08 (t, J = 9.6Hz, 2H, H-3),
3.65 (dd, J = 9.6Hz, J = 3.6Hz, 2H, H-2), 3.55 (t,
J = 9.6Hz, 2H, H-4), 3.23 (dd, J = 13.2Hz, J = 2.8Hz,
2H, H-6), 3.20 (dd, J = 13.2Hz, J = 2.8Hz, 2H, H-60);
13C NMR (68MHz, CDCl3) d 138.8 (s), 138.2 (s),
138.0 (s), 128.6 (s), 128.1 (s), 128.0 (s), 127.8 (s), 127.8
5.37. Bis(N-decyl-a-D-gluco-octopyranuronamide)ether
(YH033)
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 4.99 (s, 2H, H-1), 3.7–
3.8 (m, 4H), 3.47 (m, 2H), 3.30 (s, 2H), 3.0–3.1 (m, 4H),
2.1–2.4 (m, 8H), 1.2–1.7 (m, 32H), 0.9 (m, 6H); 13C
NMR (68MHz, CD3OD) d 175.0 (s), 94.1 (s, C-1),
74.4 (s), 73.2 (s), 72.1 (s), 71.0 (s), 39.4 (s), 32.0 (s),
31.8 (s), 29.4 (s, 2 carbons), 29.2 (s, 2 carbons), 29.0

J. Wang et al. / Bioorg. Med. Chem. 12 (2004) 6397–6413
6409
(s), 127.7 (s), 127.5 (s), 94.1 (s, C-1), 81.6 (s), 79.7 (s),
5.43. 6,60-Dioctylcarbamoylamino-6,60-dideoxy-a-D-tre-
halose (YH026)
78.5 (s), 75.7 (s), 75.3 (s), 73.2 (s), 70.7 (s), 51.2 (s);
MALDI Calcd for C54H56N6O9Na ([M+Na]+) m/e
955.4001; measure m/e 955.3980.
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 5.05 (d, J = 3.3Hz, 2H,
H-1), 3.8–3.9 (m, 2H), 3.78 (t, J = 9.2Hz, 2H, H-3), 3.4–
3.5 (m, 6H), 3.1–3.3 (m, 6H), 1.1–1.5 (m, 24H), 0.89 (t,
J = 6.9Hz, 6H, CH2CH3); 13C NMR (68MHz,
CD3OD) d 160.0 (s, NHCONH), 94.2 (s, C-1), 72.8
(s), 72.0 (s), 71.6 (s), 71.2 (s), 40.8 (s), 39.9 (s),
31.7 (s), 29.9 (s), 29.2 (s), 29.1 (s), 26.7 (s), 22.4 (s),
13.1 (s); LRFAB m/e 673 ([M+Na]+); HRFAB Calcd
for C30H58N4O11Na ([M+Na]+) m/e 673.4000; measure
m/e 673.4015.
5.41. 6,60-Diamino-2,20,3,30,4,40-hexa-O-benzyl-6,60-di-
deoxy-a-D-trehalose (26)
To a solution of 25 (0.78g, 0.84mmol) in THF, 0.1M
NaOH (0.5mL) and 1M PMe3 (2.7mL, 2.7mmol) were
added. The reaction mixture was stirred at 50°C for
3h. Lots of bubbling was observed at the beginning.
The reaction was monitored with TLC till completion.
After removal of the solvent followed by purification
with a gradient column chromatography (hexane–
EtOAc–MeOH = 60:40:0 to 35:35:30), the product was
afforded as a yellowish viscous oil (0.56g, 0.64mmol,
76%). 1H NMR (270MHz, CDCl3) d 7.10–7.50 (m,
30H), 5.20 (s, br, 2H, H-1), 5.00 (d, J = 10.9Hz, 2H,
PhCH2O), 4.87 (d, J = 10.9Hz, 2H, PhCH2O), 4.85 (d,
J = 10.9Hz, 2H, PhCH2O), 4.50–4.80 (m, 6H, PhCH2O),
2.50–4.30 (m, 12H); 13C NMR (68MHz, CDCl3) d 138.7
(s), 138.2 (s), 138.2 (s), 128.6 (s), 128.5 (s), 128.1 (s), 127.9
(s), 127.4 (s), 127.3 (s), 127.2 (s), 93.2 (s, C-1), 81.7 (s),
79.7 (s), 78.6 (s), 77.3 (s), 75.6 (s), 74.9 (s), 73.3 (s), 50.6
(s); LRFAB m/e 881 ([M+H]+); HRFAB Calcd for
C54H61N2O9 ([M+H]+) m/e 881.4377; measure m/e
881.4418.
5.44. 6,60-Dideoxy-6,60-didecylamino-2,20,3,30,4,40-hexa-
O-benzyl-a-D-trehalose (28)
To a solution of 26 (0.1g, 0.11mmol) dissolved in
CH2Cl2 and Et3N (0.040mL, 0.28mmol) at 0°C, deca-
noyl chloride (0.050mL, 0.28mmol) was added slowly.
The reaction was stirred for 12h, allowing the tempera-
ture to be warmed up to room temperature. After com-
pletion of the reaction, the reaction was quenched with
water and diluted with EtOAc. The combined organic
solution was washed with 1N HCl(aq), water, saturated
NaHCO3(aq), and brine, then dried over Na2SO4(s)
.
After removal of the solvent, followed by purification
with a gradient column chromatography (hexane–
EtOAc = 75:25 to 25:75), the product was afforded as
a viscous yellowish oil (0.12g, 0.1mmol, 89%). 1H
NMR (270MHz, CDCl3) d 7.10–7.50 (m, 30H), 5.36
(d, br, J = 5.3Hz, 2H, CONH), 5.08 (d, J = 3.6Hz,
2H, H-1), 5.00 (d, J = 10.9Hz, 2H, PhCH2O), 4.90 (d,
J = 10.9Hz, 2H, PhCH2O), 4.85 (d, J = 10.2Hz, 2H,
PhCH2O), 4.74 (d, J = 12.2Hz, 2H, PhCH2O), 4.65 (d,
J = 12.2Hz, 2H, PhCH2O), 4.62 (d, J = 10.2Hz, 2H,
PhCH2O), 4.0–4.2 (m, 4H), 3.84 (td, J = 9.6Hz,
J = 3.3Hz, 2H, H-5), 3.51 (dd, J = 9.6Hz, J = 3.3Hz,
2H, H-2), 3.32 (t, J = 9.6Hz, 2H, H-4), 3.06 (dd,
J = 13.9Hz, J = 3.3Hz, 2H, H-6), 2.00 (t, J = 7.9Hz,
4H, CH2CONH), 1.1–1.6 (m, 28H), 0.89 (t, J = 5.9Hz,
6H, CH2CH3); 13C NMR (68MHz, CDCl3) d 173.0 (s,
CONH), 138.6 (s), 138.1 (s), 138.0 (s), 128.6 (s), 128.5
(s), 128.0 (s), 127.9 (s), 127.8 (s), 127.4 (s), 94.0 (s,
C-1), 81.6 (s), 79.4 (s), 78.8 (s), 75.8 (s), 75.4 (s), 73.2
(s), 69.7 (s), 39.3 (s), 36.9 (s), 32.0 (s), 29.6 (s), 29.45
(s, 2 carbons), 29.37 (s), 25.8 (s), 22.8 (s), 14.2 (s);
MALDI Calcd for C74H96N2O11Na ([M+Na]+) m/e
1211.6906; measure m/e 1211.6946.
5.42. 6,60-Dioctylcarbamoylamino-2,20,3,30,4,40-hexa-O-
benzyl-6,60-dideoxy-a-D-trehalose (27)
To a solution of 26 (0.20g, 0.22mmol) dissolved in 5mL
of CH2Cl2 at ꢁ78°C, triphosgene (0.044g, 0.15mmol)
and Et3N (0.13mL, 0.91mmol) were added. The reaction
was allowed to warm up to room temperature. After
being stirred for another 5min, the reaction mixture
was cooled to 0°C, and was added with n-octyl amine
(0.075g, 0.46mmol). The reaction was stirred for 12h,
allowing the temperature to be warmed up to room tem-
perature. After the completion of the reaction, water was
added to quench the reaction followed by EtOAc dilu-
tion. The combined organic solution was washed with
1N HCl(aq), water, saturated NaHCO3(aq), and brine,
then dried over Na2SO4(s). After removal of the solvent
followed by purification with a gradient column chroma-
tography (hexane–EtOAc = 75:25 to 20:80), the product
was afforded as a viscous yellowish oil (0.13g, 0.11mmol,
49%). 1H NMR (270MHz, CDCl3) d 7.10–7.50 (m, 30H),
5.14 (d, J = 3.0Hz, 2H, H-1), 5.00 (d, J = 10.9Hz, 2H,
PhCH2O), 4.87 (d, J = 10.9Hz, 2H, PhCH2O), 4.85 (d,
J = 10.6Hz, 2H, PhCH2O), 4.65 (d, J = 10.6Hz, 2H,
PhCH2O), 4.65 (s, 4H, PhCH2O), 4.03–4.16 (m, 6H),
3.50 (dd, J = 9.2Hz, J = 3.0Hz, 2H, H-2), 3.40 (t,
J = 9.2Hz, 4H), 3.0–3.2 (m, 8H), 1.10–1.50 (m, 24H),
0.87 (t, J = 5.9Hz, 6H, CH2CH3); 13C NMR (68MHz,
CDCl3) d 158.4 (s, NHCONH), 138.7 (s), 138.2 (s),
138.1 (s), 128.6 (s), 128.5 (s), 128.0 (s), 127.8 (s), 127.7
(s), 127.5 (s), 93.1 (s, C-1), 81.5 (s), 79.5 (s), 78.8 (s),
75.6 (s), 75.1 (s), 72.9 (s), 70.6 (s), 41.2 (s), 40.5 (s), 31.9
(s), 30.3 (s), 29.4 (s), 29.4 (s), 27.0 (s), 22.7 (s), 14.2 (s);
MALDI Calcd for C72H94N4O11Na ([M+Na]+) m/e
1213.6811; measure m/e 1213.6889.
5.45. 6,60-Dideoxy-6,60-didecylamino-a-D-trehalose
(YH025)
Please refer to the general procedure for hydrogenation
1H NMR (270MHz, CD3OD) d 5.04 (d, J = 3.9Hz, 2H,
H-1), 3.8–3.9 (m, 2H), 3.76 (t, J = 9.2Hz, 2H, H-3), 3.4–
3.5 (m, 6H), 3.13 (t, J = 9.2Hz, 2H, H-4), 2.22 (t,
J = 7.3Hz, 4H, CH2CONH), 1.1–1.6 (m, 28H), 0.89 (t,
J = 6.9Hz, 6H, CH2CH3); 13C NMR (68MHz, CD3OD)
d 175.8 (s, CONH), 94.2 (s, C-1), 72.8 (s), 72.0 (s), 71.8
(s), 70.7 (s), 40.1 (s), 35.7 (s), 31.7 (s), 29.3 (s), 29.2 (s),
29.1 (s), 29.0 (s), 25.8 (s), 22.4 (s), 12.3 (s); LRFAB

6410
J. Wang et al. / Bioorg. Med. Chem. 12 (2004) 6397–6413
m/e 671 ([M+Na]+); HRFAB Calcd for C32H60N2O11Na
([M+Na]+) m/e 671.4095; measure m/e 671.4067.
J = 9.9Hz, 2H, H-2), 3.30 (dd, J = 9.6Hz, J = 9.2Hz,
2H, H-4), 3.09 (m, 2H, H-6), 2.51 (dd, J = 6.3Hz,
J = 5.9Hz, 4H, H-10), 1.40 (m, 4H, H-20), 1.26 (m,
20H), 0.87 (m, 6H, H-70); 13C NMR (68MHz, CDCl3)
d 171.6 (s), 138.76 (s), 138.2 (s), 137.9 (s), 128.5 (s),
128.0 (s), 127.9 (s), 127.8 (s), 127.7, 93.8 (s, C-1), 81.6
(s), 79.6 (s), 78.6 (s), 75.8 (s), 75.4 (s), 73.1 (s), 69.8 (s),
52.5 (s), 50.3 (s), 38.8 (s), 31.9 (s), 30.1 (s), 29.3 (s),
27.2 (s), 22.7 (s), 14.2 (s); MALDI Calcd for
C72H95N4O11 ([M+H]+) 1191.6992; measure 1191.7062.
5.46. 6,60-Dideoxy-6,60-di(monobromoacetyl)amino-2,20,
3,30,4,40-hexa-O-benzyl-a-D-trehalose (29)
To a solution of 26 (0.32g, 0.36mmol) in anhydrous
DMF (3mL), bromoacetic acid (7.2mL of 1M solution
in DMF), and diisopropylcarboimide (1.3mL) were
added. The reaction mixture was stirred at room tem-
perature for 4h. After complete consumption of the
starting material (monitored by TLC, EtOAc–hexa-
nes = 50:50), the reaction mixture was concentrated,
and the residue was diluted with EtOAc, followed by
washing with 1N HCl, saturated NaHCO3 solution
(three times), brine then dried over Na2SO4. After re-
moval of the solvent followed by a gradient column
chromatography (hexanes–EtOAc = 80:20 to 20:80),
the product was afforded as a yellowish oil (0.30g,
5.48. 6,60-Dideoxy-6,60-di(monoheptaminoacetyl)amino-
a-D-trehalose (YH037)
Please refer to the general procedure for hydrogenation.
Using Pd/C as catalyst instead of Pd(OH)2/C. 1H NMR
(270MHz, CD3OD) d 5.07 (d, J = 3.6Hz, 2H, H-1), 3.92
(m, 2H), 3.7–3.8 (m, 4H), 3.49 (dd, J = 3.6Hz,
J = 9.9Hz, 2H, H-2), 3.3–3.4 (m, 2H), 3.16 (dd,
J = 9.6Hz, J = 9.2Hz, 2H), 3.02 (m, 4H), 1.70 (s, 4H),
1.3–1.4 (m, 20H), 0.90 (m, 6H); 13C NMR (68MHz,
CD3OD) d 165.6 (s), 95.0 (s), 73.0 (s), 71.9 (s, 2 carbons),
70.7 (s), 58.3 (s), 41.1 (s), 40.3 (s), 31.4 (s), 28.5 (s), 26.2
(s), 25.9 (s), 22.3 (s), 13.1 (s); MALDI Calcd for
C30H59N4O11 ([M+H]+) 651.4175; measure 651.4176.
1
0.27mmol, 74%). H NMR (270MHz, CDCl3) d 7.2–
7.4 (m, 30H), 5.17 (d, J = 3.3Hz, 2H, H-1), 5.02 (d,
J = 10.9Hz, 2H, PhCH2O), 4.90 (d, J = 10.9Hz, 2H,
PhCH2O), 4.89 (d, J = 10.2Hz, 2H, PhCH2O), 4.79 (d,
J = 11.9Hz, 2H, PhCH2O), 4.69 (d, J = 11.9Hz, 2H,
PhCH2O), 4.63 (d, J = 10.2Hz, 2H, PhCH2O), 4.0–4.1
(m, 4H, H4, H-5), 3.76 (d, J = 13.5Hz, 2H, CH2Br),
3.69 (d, J = 13.5Hz, 2H, CH2Br), 3.6–3.7 (m, 2H, H-
6), 3.58 (dd, J = 9.6Hz, J = 3.3Hz, 2H, H-2), 3.34 (dd,
J = 6.6Hz, J = 9.2Hz, 2H, H-3), 3.19 (d, J = 13.9Hz,
2H, H-6); 13C NMR (68MHz, CDCl3) d 165.6 (s),
138.5 (s), 138.0 (s), 137.9 (s), 128.6 (s), 128.56 (s),
128.5 (s), 128.1 (s), 127.9 (s), 127.8 (s), 127.4 (s), 93.6
(s, C-1), 81.6 (s), 79.6 (s), 78.8 (s), 75.8 (s), 75.4 (s),
73.4 (s), 69.6 (s), 40.2 (s), 29.3 (s); MALDI Calcd for
C58H62Br2N2O11Na ([M+Na]+) 1143.2613; measure
1143.2642.
5.49. Bis((6E)-2,3,4,-tri-O-benzyl-6,7-dideoxy-a-D-gluco-
oct-6-enopyranosyluronate)ether (31)
A solution of 14 (0.20g, 0.21mmol) and LiOH–H2O
(0.03g, 0.71mmol) in THF/H2O (6mL/2mL) was stirred
at room temperature for 2days. After completion of the
reaction (monitored by TLC, EtOAc–hexane = 3:1),
Dowex 50W (H+ form) resin was added, and the reac-
tion mixture was stirred for 1h. The reaction mixture
was filtered through Celite. After removal of the sol-
vents, the crude product was purified with a gradient
column chromatography (hexane–EtOAc to EtOAc–
MeOH = 60:40 to 90:10). The purified product was ob-
tained as white solid (0.070g, 0.080mmol, 36%). 1H
NMR (270MHz, CDCl3) d 7.2–7.4 (m, 30H), 7.09 (dd,
J = 15.8Hz, J = 3.0Hz, 2H, H-6), 6.03 (d, J = 15.8Hz,
2H, H-7), 5.13 (s, 2H, H-1), 5.03 (d, J = 10.5Hz, 2H,
PhCH2O), 4.92 (d, J = 10.8Hz, 2H, PhCH2O), 4.83 (d,
J = 10.5Hz, 2H, PhCH2O), 4.6–4.8 (m, 8H, PhCH2O,
H-5), 4.08 (dd, J = 8.8Hz, J = 9.3Hz, 2H, H-3), 3.55
(d, J = 8.8Hz, 2H, H-2), 3.26 (dd, J = 9.3Hz,
J = 9.3Hz, 2H, H-4); 13C NMR (68MHz, CDCl3) d
171.7 (s, COOH), 147.1 (s), 138.6 (s), 137.7 (s), 137.6
(s), 128.7 (s), 128.6 (s), 128.5 (s), 128.2 (s), 128.0 (s),
127.8 (s), 120.7 (s), 94.4 (s, C1), 81.8 (s), 81.7 (s), 79.2
(s), 75.9 (s), 75.7 (s), 73.4 (s), 69.9 (s); MALDI Calcd
for C58H58O13Na ([M+Na]+) 985.3770; measure
985.3798.
5.47. 6,60-Dideoxy-6,60-di(monoheptaminoacetyl)amino-
2,20,3,30,4,40-hexa-O-benzyl-a-D-trehalose (30)
To a solution of 29 (0.15g, 0.13mmol) in anhydrous
DMF (5mL), heptamine (0.0mL, 0.32mmol), and
Cs2CO3 (0.10g, 0.32mmol) were added. The reaction
mixture was stirred overnight at room temperature.
The reaction can be monitored by TLC. The product
has a Rf of 0 when eluted with EtOAc–MeOH (9:1) solu-
tion and a Rf of 0.3 when eluted with i-PrOH–1M
NH4OH (8:1) solution. After completion of the reaction,
the reaction mixture was concentrated and loaded to a
short column (5cm in height) packed with TLC silica
gel on top of Celite. The column was eluted with a series
of solutions as following: CH2Cl2, CH2Cl2–EtOAc,
EtOAc, EtOAc–MeOH. The fractions containing de-
sired product were analyzed by TLC and collected.
The product was afforded as white solid (0.12g,
1
0.11mmol, 75%). H NMR (270MHz, CDCl3) d 7.2–
5.50. Bis(octanoyl (6E)-2,3,4,-tri-O-benzyl-6,7-dideoxy-
a-D-gluco-oct-6-enopyranosyluronoylhydrazine)ether (32)
7.4 (m, 30H), 5.14 (d, J = 3.6Hz, 2H, H-1), 4.97 (d,
J = 10.6Hz, 2H, PhCH2O), 4.87 (d, J = 10.6Hz, 2H,
PhCH2O), 4.83 (d, J = 10.2Hz, 2H, PhCH2O), 4.71 (d,
J = 11.9Hz, 2H, PhCH2O), 4.65 (d, J = 11.9Hz, 2H,
PhCH2O), 4.63 (d, J = 10.2Hz, 2H, PhCH2O), 4.13
(m, 2H, H-6), 4.06(dd, J = 9.9Hz, J = 9.6Hz, 2H,
H-3), 3.8–3.9 (m, 2H, H-5), 3.50 (dd, J = 3.6Hz,
To a solution of 31 (0.15g, 0.16mmol) in anhydrous
DMF (5mL), octanoic hydrazine (0.10g, 85% purity,
0.49mmol), and EDC (0.10g, 0.52mmol) were added.
The reaction mixture was stirred overnight at room tem-
perature. After the complete consumption of starting

J. Wang et al. / Bioorg. Med. Chem. 12 (2004) 6397–6413
6411
material (monitored by TLC, EtOAc–hexanes = 3:1),
the reaction mixture was concentrated and the residue
was diluted with CH2Cl2. To this mixture, NaHCO3(s)
and Na2CO3(s) were added. After being stirred for 1h,
the reaction mixture was filtered through Celite. After
removal of the solvents followed by a gradient column
chromatography (hexanes–EtOAc = 80:20 to 20:80),
the product was afforded as white solid (0.080g,
0.070mmol, 41%). 1H NMR (270MHz, CDCl3) d
10.46 (d, J = 4.9Hz, 2H), 10.06 (s, 2H), 7.2–7.4 (m,
30H), 6.96 (dd, J = 15.5Hz, J = 5.3Hz, 2H, H-6), 6.00
(d, J = 15.5Hz, 2H, H-7), 5.28 (d, J = 3.0Hz, 2H, H-
1), 4.93 (d, J = 10.9Hz, 2H, PhCH2O), 4.83 (d,
J = 10.9Hz, 2H, PhCH2O), 4.5–4.7 (m, 8H, H5,
PhCH2O), 4.43 (d, J = 10.6Hz, 2H, PhCH2O), 4.04
(dd, J = 10.2Hz, J = 9.2Hz, 2H, H-4), 3.47 (dd,
J = 3.0Hz, J = 9.6Hz, 2H, H-2), 3.19 (dd, J = 9.6Hz,
J = 9.2Hz, 2H, H-3), 2.36 (dd, J = 7.3Hz, J = 7.6Hz,
4H), 1.2–1.3 (m, 20H), 0.88 (m, 6H); 13C NMR
(68MHz, CDCl3) d 168.8 (s), 160.4 (s), 141.4 (s, C6),
138.7 (s), 138.0 (s), 137.8 (s), 128.6 (s), 127.6 (s), 127.3
(s), 128.3 (s), 128.0 (s), 127.8 (s), 127.7 (s), 127.6 (s),
127.3 (s), 121.6 (s, C7), 95.1 (s, C-1), 82.0 (s), 81.3 (s),
80.0 (s), 75.6 (s), 75.2 (s), 72.8 (s), 70.7 (s), 34.0 (s),
31.8 (s), 29.3 (s), 29.1 (s), 25.7 (s), 22.7 (s), 14.2
(s); MALDI Calcd for C74H90N4O13Na ([M+Na]+)
1265.6397; measure 1265.6365.
PhCH2O), 4.48 (d, J = 10.9Hz, 1H, PhCH2O), 4.40
(d, J = 11.9Hz, 1H, PhCH2O), 4.0–4.2 (m, 3H), 3.69
(dd, J = 9.9Hz, J = 9.2Hz, 1H), 3.5–3.6 (m, 7H), 3.39
(d, J = 10.6Hz, 1H); 13C NMR (68MHz, CDCl3) d
138.96 (s), 138.90 (s), 138.44 (s), 138.35 (s), 138.25 (s),
138.21 (s), 137.9 (s), 128.56 (s), 128.47 (s), 128.44
(s), 128.2 (s), 128.08 (s), 128.06 (s), 127.98 (s), 127.78
(s), 127.72 (s), 127.62 (s), 127.5 (s), 94.4 (s), 94.2 (s),
81.89 (s), 81.68 (s), 79.64 (s), 79.45 (s), 77.8 (s), 77.45
(s), 75.70 (s, 2 carbons), 75.16 (s, 2 carbons), 73.61 (s),
73.0 (s), 72.92 (s), 71.3 (s), 70.8 (s), 68.3 (s), 61.6 (s);
MALDI Calcd for C61H64O11Na ([M+Na]+) 995.4341;
measure 995.4300.
5.53. 60-Azido-2,20,3,30,4,40,6-hepta-O-benzyl-6,60-dide-
oxy-a-D-trehalose (35)
To a solution of 34 (0.53g, 0.55mmol) in anhydrous
pyridine (10mL), TsCl (1.2g, 6.5mmol) was added.
After being stirred for 3h, the reaction was quenched
by addition of water. The reaction mixture was diluted
with EtOAc, and washed with 1N HCl (three times),
water, saturated NaHCO3 solution, brine then dried
over Na2SO4. After removal of the solvent, the crude
product was redissolved in DMF (10mL), and added
with excess NaN3 (ca. 0.5g). The reaction mixture was
stirred at 100°C for 5h then was concentrated. The
brownish solid was added with EtOAc and filtered
through Celite. After removal of the solvents, followed
by purification with a gradient column chromatography
(hexane–EtOAc = 75:25 to 25:75), the product was
afforded as a clear oil (0.46g, 0.46mmol, 84%). 1H
NMR (270MHz, CDCl3) d 7.1–7.4 (m, 35H), 5.22 (d,
J = 3.6Hz, 2H, H-1, H-10), 5.00 (d, J = 10.9Hz, 1H,
PhCH2O), 4.99 (d, J = 10.9Hz, 1H, PhCH2O), 4.7–4.9
(m, 8H, PhCH2O), 4.57 (d, J = 11.2Hz, 1H, PhCH2O),
4.55 (d, J = 12.2Hz, 1H, PhCH2O), 4.47 (d,
J = 10.6Hz, 1H, PhCH2O), 4.39 (d, J = 11.9Hz, 1H,
PhCH2O), 3.9–4.2 (m, 5H), 3.69 (dd, J = 9.9Hz,
J = 9.2Hz, 1H), 3.4–3.6 (m, 5H), 3.18 (d, J = 3.3Hz,
1H); 13C NMR (68MHz, CDCl3) d 138.91 (s), 138.75
(s), 138.40 (s), 138.24 (s), 138.15 (s, 2 carbons), 137.90
(s), 128.54 (s), 128.47 (s), 128.43 (s), 128.06 (s),
127.95 (s), 127.77 (s), 127.70 (s), 127.63 (s), 127.53 (s),
127.44 (s), 94.6 (s), 94.1 (s), 81.9 (s), 81.5 (s), 79.6
(s), 79.5 (s), 78.4 (s), 77.8 (s), 75.7 (s, 2 carbons), 75.25
(s), 75.17 (s), 73.6 (s), 73.1 (s), 72.9 (s), 70.9 (s), 70.4 (s),
68.2 (s), 51.2 (s); MALDI Calcd for C61H63N3O10Na
([M+Na]+) 1020.4406; measure 1020.4357.
5.51. Bis(octanoyl (a-D-gluco-oct-pyranosyluronoylhydr-
azine)ether (YH036)
Please refer to the general procedure for hydrogenation.
1H NMR (270MHz, CD3OD) d 5.04 (d, J = 3.3Hz, 2H,
H-1), 3.78 (m, 2H, H-5), 3.73 (dd, J = 9.6Hz, J = 8.9Hz,
2H, H-4), 3.48 (dd, J = 3.6Hz, J = 9.2Hz, 2H, H-2),
3.08 (dd, J = 8.9Hz, J = 9.2Hz, 2H, H-3), 2.7–2.8 (m,
4H), 2.24 (dd, J = 7.2Hz, J = 7.2Hz, 4H), 1.64 (m,
4H), 1.31 (m, 20H), 0.89 (m, 6H); 13C NMR (68MHz,
CD3OD) d 173.9 (s), 173.6 (s), 94.1 (s), 74.5 (s), 73.1
(s), 72.0 (s), 70.7 (s), 33.5 (s), 31.6 (s), 29.6 (s), 28.9 (s),
28.8 (s), 27.1 (s), 25.3 (s), 22.4 (s), 13.1 (s); MALDI
Calcd for C32H58N4O13Na ([M+Na]+) 729.3893; meas-
ure 729.3930.
5.52. 2,20,3,30,4,40,6-Hepta-O-benzyl-a-D-trehalose (34)
To a solution of 33 (1.30g, 1.2mmol) in anhydrous
CH2Cl2 (20mL) and Ac2O (0.15mL, 1.6mmol) at
ꢁ78°C, TMSOTf (0.31mL, 1.6mmol) was slowly
added. The reaction was stirred for 9h, allowing the
temperature to be warmed up to room temperature. A
solution of NaOMe (3mL, 2M in MeOH) was added.
After being stirred for 0.5h, Amberlite IR 120 (H+ form)
was added. The reaction mixture was filtered and con-
centrated. After purification with a gradient column
chromatography (hexane–EtOAc = 75:25 to 25:75), the
product was afforded as a clear oil (0.59g, 0.61mmol,
50%) along with the recovered 33 (0.31g, 0.29mmol,
24%). 1H NMR (270MHz, CDCl3) d 7.1–7.4 (m,
35H), 5.21 (d, J = 3.0Hz, 1H, H-1), 5.20 (d,
J = 3.3Hz, 1H, H-10), 5.01 (d, J = 10.9Hz, 2H,
PhCH2O), 4.7–4.9 (m, 8H, PhCH2O), 4.66 (d,
J = 11.2Hz, 1H, PhCH2O), 4.56 (d, J = 12.2Hz, 1H,
5.54. 60-Azido-2,20,3,30,4,40-hexa-O-benzyl-60-deoxy-a-D-
trehalose (36)
Please refer to the procedure for the preparation of 34.
1H NMR (270MHz, CDCl3) d 7.1–7.4 (m, 30H), 5.18
(d, J = 3.0Hz, 1H, H-1), 5.17 (d, J = 3.3Hz, 1H, H-10),
5.01 (d, J = 11.2Hz, 1H, PhCH2O), 5.00 (d,
J = 10.9Hz, 1H, PhCH2O), 4.7–4.9 (m, 8H, PhCH2O),
4.65 (d, J = 11.2Hz, 1H, PhCH2O), 4.57 (d,
J = 10.9Hz, 1H, PhCH2O), 3.9–4.2 (m, 5H), 3.5–3.6
(m, 6H), 3.18 (d, J = 3.3Hz, 1H); 13C NMR (68MHz,
CDCl3) d 138.8 (s), 138.7 (s), 138.28 (s), 138.16 (s, 2 car-
bons), 138.03 (s), 128.55 (s), 128.50 (s), 128.18 (s), 128.04

6412
J. Wang et al. / Bioorg. Med. Chem. 12 (2004) 6397–6413
(s), 127.94 (s), 127.75 (s), 127.68 (s), 127.57 (s), 127.51
(s), 94.2 (s), 94.0 (s), 81.7 (s), 81.5 (s), 79.6 (s, 2 carbons),
78.4 (s), 77.5 (s), 75.7 (s, 2 carbons), 75.3 (s), 75.2 (s),
73.17 (s), 73.09 (s), 71.4 (s), 70.5 (s), 61.6 (s), 51.2 (s);
MALDI Calcd for C54H57N3O10Na ([M+Na]+)
930.3936; measure 930.3934.
3.9–4.1 (m, 4H), 3.5–3.6 (m, 3H), 3.2–3.3 (m, 4H), 1.48
(dd, J = 6.9Hz, J = 6.9Hz, 2H), 1.2–1.3 (m, 10H), 0.87
(dd, J = 6.9Hz, J = 6.9Hz, 3H); 13C NMR (68MHz,
CDCl3) d 164.9 (s), 149.1 (s), 139.4 (s), 138.7 (s, 2
carbons), 138.3 (s), 138.1 (s), 137.9 (s), 128.5 (s), 128.4
(s), 128.0 (s), 127.7 (s), 127.5 (s), 127.4 (s), 125.1 (s),
94.1 (s, C-1), 93.9 (s), 82.2 (s), 81.5 (s, 2 carbons), 79.7
(s), 79.3 (s), 78.4 (s), 75.8 (s), 75.7 (s), 75.6 (s), 75.3 (s),
73.2 (s), 72.9 (s), 70.6 (s), 70.5 (s), 51.1 (s), 39.6
(s), 31.9 (s), 29.7 (s), 29.4 (s), 29.3 (s), 27.0 (s), 22.7 (s),
14.2 (s); MALDI Calcd for C64H74N4O10Na
([M+Na]+) 1081.5297; measure 1081.5306.
5.55. 6-Azido-2,3,4-tri-O-benzyl-6-deoxy-a-D-trehalose
methyl (6E)-20,30,40-tri-O-benzyl-60,70-dideoxy-a-D-gluco-
oct-6-enopyranosyluronate)ether (37)
To a solution of (COCl)2 (29lL, 0.33mmol) in 5mL
anhydrous CH2Cl2 at ꢁ78°C, anhydrous DMSO
(47lL, 0.66mmol) was added and the resulting solution
was stirred for 15min, allowing the temperature to be
warmed up to ꢁ65°C. To the reaction flask, a solution
of 36 (0.15g, 0.17mmol) in 5mL CH2Cl2 was added.
The reaction mixture was stirred for 0.5h allowing the
temperature to be warmed up to ꢁ45°C. To this solu-
tion, anhydrous DIPEA (0.23mL, 1.32mmol) was
added, and the reaction was allowed to be warmed up
to ꢁ10–0°C in 0.5h. After completion of the reaction,
the reaction mixture quenched with 1N HCl and diluted
with CH2Cl2. The organic layer was washed with pH7
buffer (three times), brine, and dried over Na2SO4. After
removal of the solvents, the product, aldehyde, was used
for the next step without further purification. To a solu-
tion of methyl diethylphosphonoacetate (50lL,
0.27mmol) in THF (5mL) at 0°C, NaH (0.011g,
0.27mmol) was added. After being stirred for 1h, the
aldehyde from first step was added. The reaction was
stirred for another 12h. After completion of the reaction,
50lL MeOH was added to quench the reaction. After re-
moval of the solvents, the residue was dissolved in
EtOAc, and washed with brine then dried over Na2-
SO4(s). After removal of the solvent, followed by purifica-
tion with a gradient column chromatography (hexane–
EtOAc = 90:10 to 50:50), the product was obtained as
5.57. 6-Amino-6-deoxy-a-D-trehalose (N-octyl-a-D-gluco-
octopyranosyluronamide)ether (YH041)
Please refer to the general procedure for hydrogenation
1H NMR (400MHz, D2O) d 5.13 (d, J = 3.6Hz, 1H, H-
1), 5.09 (d, J = 3.8Hz, 1H, H-1), 3.6–4.0 (m, 5H), 3.1–
3.3 (m, 5H), 2.2–2.4 (m, 2H), 2.10 (m, 1H), 1.71 (m,
1H), 1.44 (m, 2H), 1.23 (m, 12H), 0.81 (m, 3H); 13C
NMR (100MHz, D2O) d 175.1 (s), 94.0 (s), 93.8 (s),
73.6 (s), 72.7 (s), 72.4 (s), 71.8 (s), 71.6 (s), 71.5
(s), 71.0 (s), 68.4 (s), 40.8 (s), 39.7 (s), 32.3 (s), 31.3 (s),
28.6 (s, 2 carbons), 28.5 (s), 27.3 (s), 26.3 (s), 22.3 (s),
13.7 (s); MALDI Calcd for C22H42N2O10Na
([M+Na]+) 517.2732; measure 517.2749.
5.58. Procedure for disk assay of antimicrobial activity
Disk diffusion assays were modified from methods out-
lined by the National Committee on Clinical Laboratory
Standards.15 M. smegmatis (ATCC 14468) was inocu-
lated in LB (Lennox) broth, and the resulting solution
was incubated at 37°C for 2days then was transferred
into the LB (Lennox) agar plate by swab. Aliquots
(20lL) of solutions of INH (250lg/mL) and trehalose
derivatives (10mg/mL) were applied to the disks. The
plate was incubated at 37°C for 2–3days. The diameter
of the zone of inhibition is measured, to the nearest
whole millimeter.
1
a pale yellowish oil (0.16g, 0.17mmol, 99%). H NMR
(270MHz, CDCl3)
d
7.3–7.4 (m, 30H), 6.94
(dd, J = 15.8Hz, J = 4.3Hz, 1H), 6.01 (dd, J = 15.8Hz,
J = 1.6Hz, 1H), 5.20 (d, J = 3.6Hz, 1H, H-1), 5.10 (d,
J = 3.6Hz, 1H, H-1), 4.5–5.1 (m, 13H), 3.9–4.1 (m,
4H), 3.75 (s, 3H, COOCH3), 3.4–3.6 (m, 3H), 3.2–3.3
(m, 2H); 13C NMR (100MHz, CDCl3) d 166.8 (s),
144.7 (s), 138.9 (s), 138.8 (s), 138.3 (s), 138.2 (s), 137.9
(s), 137.8 (s), 129.1 (s), 128.9 (s), 128.63 (s), 128.58 (s),
128.49 (s), 128.2 (s), 128.1 (s), 127.9 (s), 127.85 (s),
127.6 (s), 121.3 (s), 94.4 (s), 94.3 (s, C-1), 82.2 (s), 81.74
(s), 81.66 (s), 79.6 (s, 2 carbons), 78.5 (s), 75.9 (s), 75.8
(s), 75.7 (s), 75.4 (s), 73.3 (s, 2 carbons), 70.7 (s), 70.1
(s), 51.8 (s), 51.3 (s); MALDI Calcd for C57H59N3O11Na
([M+Na]+) 984.4042; measure 984.3991.
Acknowledgements
We thank the financial support from the Utah State
University (New Faculty Research Grant and CURI
grant) and the support from Department of Chemistry
and Biochemistry, Utah State University. We also thank
Prof. Lisa Berreau for assistance on editing the
manuscript.
References and notes
5.56. 6-Azido-2,3,4-tri-O-benzyl-6-deoxy-a-D-trehalose
((6E)-20,30,40-tri-O-benzyl-60,70-dideoxy-N-octyl-a-D-
gluco-oct-6-enopyranosyluronamide)ether (38)
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