Synthesis of triazole-linked β-C-glycosyl dimers as inhibitors of PTP1B

Article abstract of DOI:10.1016/j.bmc.2008.09.066

Protein tyrosine phosphatase 1B (PTP1B) has emerged as a promising target for type 2 diabetes. We have successfully synthesized dimeric acetylated and benzoylated β-C-d-glucosyl and β-C-d-galactosyl 1,4-dimethoxy benzenes or naphthalenes by click chemistr

Full text of DOI:10.1016/j.bmc.2008.09.066

Bioorganic & Medicinal Chemistry 16 (2008) 9757–9763
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
Synthesis of triazole-linked b-C-glycosyl dimers as inhibitors of PTP1B
a,
Li Lin a,b, Qiang Shen c, Guo-Rong Chen b, , Juan Xie
*
*
a PPSM, ENS Cachan, CNRS, UniverSud, 61 av President Wilson, F-94230 Cachan, France
b Laboratory for Advanced Materials and Institute of Fine Chemicals, East China University of Science and Technology, Shanghai 200237, PR China
c National Center for Drug Screening, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, PR China
a r t i c l e i n f o
a b s t r a c t
Article history:
Protein tyrosine phosphatase 1B (PTP1B) has emerged as a promising target for type 2 diabetes. We have
Received 20 August 2008
Revised 22 September 2008
Accepted 26 September 2008
Available online 30 September 2008
successfully synthesized dimeric acetylated and benzoylated b-C-
dimethoxy benzenes or naphthalenes by click chemistry. These compounds were further transformed
into the corresponding b-C- -glycosyl-1,4-quinone derivatives by CAN oxidation. The in vitro inhibition
test showed that dimeric benzoylated b-C- -glycosyl 1,4-dimethoxybenzenes or 1,4-benzoquinones were
D-glucosyl and b-C-D-galactosyl 1,4-
D
D
good inhibitors of PTP1B (IC50: 0.62–0.88
derivatives.
l
M), with no significant difference between gluco and galacto
Keywords:
Protein tyrosine phosphatase 1B
Diabetes
Ó 2008 Elsevier Ltd. All rights reserved.
Aryl C-glycosyl compounds
Click chemistry
1,4-Benzoquinone
1,4-Naphthoquinone
1. Introduction
(compound I, Fig. 1) showed a good inhibition against PTP1B
(IC50 = 4.85
M).19 This result prompted us to prepare other C-glyco-
l
Protein tyrosine phosphatase 1B (PTP1B) is an important regula-
tor of tyrosine kinase receptor-mediated responses, and influences
negatively insulin sensitivity.1–3 Indeed, PTP1B knockout mice dis-
play increased insulin sensitivity and tyrosine phosphorylation of
the insulin receptor.4,5 These results attracted considerable interest
for the development of pharmacological PTP1B inhibitors for the
treatment of insulin resistance, in particular non-insulin-dependent
diabetes mellitus (type 2 diabetes),6,7 which is characterized by a
deficient insulin cascade and is therefore also termed insulin resis-
tance(IR). Uptodate, varioustherapeuticstrategieshavebeendevel-
oped for targeting PTP1B in diabetes.8–18 Inhibitors of PTP1B
generally fall into two categories: non-hydrolysable phosphotyro-
sine mimetics (or affinity-based) and covalent inactivators. The first
category has received the most attention; numerous phosphotyro-
sinemimeticshavebeenreported, includingdifluoromethylphosph-
onates, cinnamic acid, oxalylamino benzoic acid, isoxazole
syl quinone derivatives as new sugar-based small molecule inhibi-
tors of PTP1B. Two naturally occurring allosteric binding sites have
recently been identified in the PTP1B, which are located ꢀ8 and
ꢀ20 Å, respectively, from the catalytic site.20–22 Potent bidentate
inhibitorsofPTP1Bhavebeendevelopedbasedonthisstructuralfea-
ture.8–13,23,24 We then decided to prepare dimeric b-C-aryl glyco-
sides as well as their quinones derivatives as potential bidentate
inhibitors of PTP1B.
O
OAc
O
AcO
AcO
OAc
O
I (IC50 = 4.85 μM)
OMe
N
O
O
N
N
N
N
N
carboxylicacid, salicyclicacid, a-ketocarboxylicacid, etc. Thesecond
O
O
O
O
O
O
O
O
RO
RO
RO
RO
category of inhibitors consists largely of reactive molecules which
modify the active site Cys215, like naphthoquinone as Michael
acceptor or oxidant, and halomethyl ketones as alkylating agents.
Recently, we found that b-C-glucopyranosyl-1,4-benzoquinone
OR
OR
OMe
OMe
N
N
N
N
O
N
N
O
O
O
O
RO
RO
RO
RO
OR
OR
O
OMe
* Corresponding authors. Tel.: +86 21 64253016; fax: +86 21 64252758
R = Ac or Bz
III
(G.-R. Chen); tel.: +33
1 47 40 55 86; fax: +33 1 47 40 24 54 (J. Xie).
II
E-mail addresses: mrs_guorongchen@ecust.edu.cn (G.-R. Chen), joanne.xie@
ppsm.ens-cachan.fr (J. Xie).
Figure 1. Structure of PTP1B inhibitors.
0968-0896/$ - see front matter Ó 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2008.09.066

9758
L. Lin et al. / Bioorg. Med. Chem. 16 (2008) 9757–9763
The recently developed ‘click chemistry’ or Cu(I)-catalysed
glycosides 18, 19, 22, and 23 were obtained in more than 86% yield
(Scheme 2). These compounds have been fully characterized by
NMR and mass spectrometry. Mild oxidation with CAN in a
mixture of acetonitrile and water converted these 1,4-dimethoxy
aromatic derivatives into the corresponding 1,4-benzoquinone or
1,4-naphthoquinone derivatives 20, 21, 24, and 25.
Various dimeric compounds have been evaluated as PTP1B
inhibitors and biological results are shown in Table 1. Benzoylat-
ed b-C-glycosyl 1,4-dimethoxybenzenes or 1,4-benzoquinones
(compounds 18–21) showed good inhibition against PTP1B, with
approximately the same IC50 value in submicromolar range. These
dimeric compounds were slightly better inhibitors than the par-
Huisgen 1,3-dipolar cycloadditions of terminal alkynes with organ-
ic azides were shown to be very effective and versatile and have
been receiving attention as a highly efficient and regioselective
reaction associated with excellent functional groups compatibil-
ity.25,26 In addition, the 1,2,3-triazole ring is resistant to hydrolysis,
oxidation, reduction, or other modes of cleavage. These important
features have allowed the synthesis of complex molecules includ-
ing dendrimers, bioconjugates, functionalized polymers and sugar
derivatives.27–30 Synthesis of PTP1B inhibitors with click chemistry
has also been reported.24,31 However, dimeric C-glycosyl com-
pounds have never been designed as inhibitors of PTP1B. Herein,
we reported the synthesis of dimeric b-C-glucosyl or b-C-galactosyl
derivatives via click chemistry (compounds II and III, Fig. 1) and
their inhibitory activity towards PTP1B.
ent monomer I (IC50 = 4.85 lM). There is no difference between
gluco and galacto derivatives (18 vs 19 and 20 vs 21). Further-
more, 1,4-dimethoxybenzene derivatives are as potent as 1,4-
benzoquinone derivatives (18 vs 20 and 19 vs 21). However,
compounds bearing 1,4-dimethoxynaphthalene or 1,4-naphtho-
quinone moiety (compounds 22–25) showed very weak inhibition
against PTP1B. Larger substituents at the anomeric position
seemed to hamper the interaction with the enzyme. Meanwhile,
benzoyl protecting groups seem to contribute to the PTP1B inhib-
itory activity, probably by binding in a hydrophobic pocket of
enzyme.
In summary, dimeric acetylated and benzoylated b-C-aryl glu-
co- and galacto-pyranosides have been successfully prepared by
Huisgen 1,3-dipolar cycloaddition reaction between 6-azido-b-C-
aryl glycosides and dipropargyl isophthalate in good yield. Mild
oxidation converted b-C-glycosyl-1,4-dimethoxy benzenes or
naphthalenes into the corresponding b-C-glycosyl 1,4-benzoqui-
none or 1,4-naphthoquinone derivatives. Benzoylated glucosyl
and galactosyl dimers containing a 1,4-dimethoxybenzene or 1,4-
benzoquinone moiety showed submicromolar inhibitory activity
2. Results and discussion
Dimeric b-C-aryl glycosides II (Fig. 1) can be obtained by Huis-
gen 1,3-dipolar cycloaddition between 6-azido-6-deoxy-b-C-glyco-
sides with dipropargyl isophthalate. Firstly, we have prepared
6-azido functionalized b-C-aryl glycosides (Scheme 1). The per-
acetylated gluco and galacto b-C-glycosides 132 and 532 were first
deacetylated under Zemplén condition and tritylated at 6-position
to furnish the compounds 2 and 6, followed by addition of BzCl to
protect the secondary hydroxyl groups as benzoyl ester in one-pot.
Detritylation with TFA gave the 6-OH free b-C-aryl glycosides 3 and
7 which were converted into the 6-azido-b-C-aryl glycosides 4 and
8 through activation as mesylate followed by nucleophilic substi-
tution with NaN3. For the synthesis of acetyl protected b-C-gluco-
syl and b-C-galactosyl-1,4-dimethoxynaphthalene derivatives 12
and 16, TBS has been used to selectively protect the 6-OH group
of b-C-glycosyl-1,4-dimethoxynaphthalene 933 and 1333 to com-
pounds 10 and 14. Subsequent desilylation with TBAF led however
to a mixture of 6-OH and 6-OAc products resulting from intramo-
lecular transesterification reaction. We then decided to realize the
desilylation under acidic condition (AcCl in MeOH). The gluco
derivative 11 can be obtained in excellent yield. However, the yield
of desilylation is only 38% for the galacto derivative 15. The yield
for 15 became worse when trityl was used as the protecting group.
The desired compounds 12 and 16 were obtained after mesylation
followed by azide substitution in good yields.
(IC50: 0.62 to 0.88 lM) against PTP1B, with no significant differ-
ence between gluco and galacto derivatives.
3. Experimental
3.1. Synthesis
Solvents were purified by standard procedures. 1H and 13C NMR
spectra were recorded on a Bruker AM-500, AC-300 or AGH-250
spectrometers in CDCl3 solutions. Optical rotations were measured
using a Perkin-Elmer 241 polarimeter at room temperature and a
10-cm 1-mL cell. Column chromatography was performed on E.
Merck Silica Gel 60 (230ꢁ400 mesh). Analytical thin-layer chroma-
tography was performed on E. Merck aluminum percolated plates
of Silica Gel 60F-254 with detection by UV and by spraying with
6 N H2SO4 and heating about 2 min at 300 °C. High resolution mass
spectra (HRMS) were recorded on a MA1212 instrument using
standard conditions (ESI, 70 eV).
We then studied the Huisgen 1,3-dipolar cycloaddition reaction
between 6-azido-b-C-aryl glycosides and dipropargyl isophthalate
17 which can be easily prepared by treatment of isophthalic
chloride with propargylic alcohol (Scheme 2). Click reactions were
realized with CuSO4 and sodium ascorbate in a mixture of dichlo-
romethane and water at room temperature. Reaction of dipropar-
gyl isophthalate 17 with aryl b-C-glucosides 4 and 12, or aryl
b-C-galactosides 8 and 16 worked very well. The dimeric b-C-aryl
OMe
OMe
OMe
OMe
R
R
1
R
R1
1
1 OTr
O
OH
O
N
OAc
O
3
i, ii, iii
iv
v, vi
R
R
O
R
R2
2
2
2
BzO
BzO
BzO
AcO
OBz
OBz
OAc
OMe
OBz
OMe
OMe
OMe
2
1: R1 = H, R2 = OAc
: R1 = OAc, R2 = H
: R1 = H, R2 = OBz, 61 %
6: R1 = OBz, R2 = H, 96 %
3
4: R1 = H, R2 = OBz, 57 %
: R1 = H, R2 = OBz, 79 %
7: R1 = OBz, R2 = H, 56 %
5
8
: R1 = OBz, R2 = H, 78 %
OMe
OMe
R
OMe
OMe
R
R1
1 OTBS
R
1
OH
OH
O
1
N
vii,viii
3
ix
v, vi
O
O
R2
R
2
R
O
2
R
2
HO
AcO
AcO
AcO
OH
OAc
OAc
OMe
OAc
OMe
9: R1 = H, R2 = OH
13
OMe
OMe
10
: R1 = H, R2 = OAc, 49 %
14: R1 = OAc, R2 = H, 65 %
12: R1 = H, R2 = OAc, 83 %
16
11
15
: R1 = H, R2 = OAc, 99 %
: R1 = OAc, R2 = H, 38 %
: R1 = OH, R2 = H
: R1 = OAc, R2 = H, 67 %
Scheme 1. Reagents and conditions: (i) MeONa, MeOH; (ii) TrCl, Pyr.; (iii) BzCl; (iv) TFA, wet CH2Cl2; (v) MsCl, TEA; (vi) NaN3, DMF, 90 °C; (vii) TBSCl, Pyr., DMAP; (viii) Ac2O;
(ix) AcCl, MeOH.

L. Lin et al. / Bioorg. Med. Chem. 16 (2008) 9757–9763
9759
i
+
Cl HO
Cl
O
O
N
O
O
O
O
17
: 72 %
N
N
N
OMe
O
N
N
O
O
O
O
O
O
O
O
BzO
BzO
BzO
BzO
ii
iii
+
17 4 or 8
OBz
OBz
OMe
OMe
O
O
N
N
N
N
O
O
N
N
O
O
BzO
BzO
BzO
BzO
OBz
: gluco 94 %
19
: galacto 90 %
OBz
20
: gluco 95 %
21
: galacto 94 %
OMe
OMe
O
O
18
N
N
N
N
N
N
O
O
O
O
O
O
O
O
AcO
AcO
AcO
AcO
ii
iii
OAc
OAc
+
17 12 or 16
OMe
OMe
O
O
N
N
N
N
O
O
N
N
O
O
AcO
AcO
AcO
AcO
OAc
OAc
OMe
O
22
23
24: gluco 90 %
25: galacto 56%
: gluco 86 %
: galacto 95 %
Scheme 2. Reagents and conditions: (i) Et3N, DMAP; (ii) CuSO4, Na ascorbate, CH2Cl2/H2O; (iii) CAN, H2O/MeCN.
(c = 0.6, CHCl3); mp = 79–82 °C; 1H NMR (300 MHz, CDCl3):
d = 8.13–7.07 (m, 31H, H-Ar), 6.79 (dd, J5,3 = 2.9 Hz, J5,6 = 8.8 Hz,
1H, H-5, dimethoxybenzene), 6.64 (d, J6,5 = 8.8 Hz, 1H, H-6, dime-
Table 1
In vitro PTP1B enzyme inhibitory activity for compounds 18–25
Compound
Inhibitiona (%)
IC50 (lM)
thoxybenzene), 5.92 (m, 2H, H-30, H-40), 5.70 (dd, J2 ,1 = 10.3 Hz,
0
0
18
19
20
21
22
23
24
25
97.8
98.5
95.8
95.5
12.5
7.5
0.87 0.04
0.88 0.01
0.62 0.06
0.72 0.05
—
—
—
—
J2 ,3 = 8.8 Hz, 1H, H-20), 5.30 (d, J1 ,2 = 9.6 Hz, 1H, H-10), 4.06 (m,
0
0
0
0
1H, H-50), 3.78, 3.58 (2s, 6H, 2 ꢂ OMe), 3.41 (d, J6 a,6 b = 10.7 Hz,
0
0
1H, H-60a), 3.18 (dd, J6’b,5 = 4.4 Hz, J6 b,6 a = 10.3 Hz, 1H, H-60b);
13C NMR (100 MHz, CDCl3): d = 166.1, 165.9, 164.7 (C@O), 153.7,
151.5 (C-1, C-4, dimethoxybenzene), 146.8, 133.6, 133.1, 132.9,
130.1, 129.9, 129.7, 129.1, 128.8, 128.5, 128.2, 128.1, 127.9,
127.8, 127.7, 127.2 (C-Ar), 125.4, 115.4, 112.8, 111.5 (C-2, C-3, C-
5, C-6, dimethoxybenzene), 79.3, 76.7, 74.6, 73.8, 73.2 (C-10 to C-
50), 61.0 (C-60), 55.9, 55.8 (2 ꢂ OMe). HRESIMS: calcd for
C54H46O10Na: 877.2989; found: m/z 877.2979.
0
0
0
8.5
0.8
a
Values tested at 20
l
g/mL concentration.
3.1.1. General procedure for the tritylation and tribenzoylation
3.1.1.2. 2-(20,30,40-Tri-O-benzoyl-60-O-triphenylmethyl-b-
D-galac-
topyranosyl)-1,4-di-methoxybenzene (6). From compound 5
(440 mg, 0.94 mmol), column chromatography (5:1 to 3:1, petro-
leum ether/EtOAc) afforded 6 as a white solid (764 mg, 96%).
Compounds 2 and 6 were synthesized by this procedure. The
known 2,3,4,6-tetra-O-acetyl-b-D-glycopyranosyl-1,4-dimethoxy-
benzene (1 or 5)32 was dissolved in MeOH, and 1–2 drops of a
1 M methanolic MeONa soln was added. The reaction mixture
was kept at rt until completion of the reaction (TLC 10:1
CH2Cl2/MeOH). Resin (H+ form) was then added to remove so-
dium ion. Filtration and concentration afford the fully deprotec-
ted glycosides which were used directly without purification.
The deprotected compound was dissolved in dry pyridine
(5 mL) and triphenylmethyl chloride (2.7 equiv) was added. The
reaction mixture was stirred at rt until TLC indicated the disap-
pearance of the starting materials, cooled to 0 °C and then BzCl
(4 equiv) was added dropwise. The reaction mixture was stirred
at rt for 6–10 h, H2O (10–15 mL) was added to quench the reac-
tion. The organic layer was diluted with CH2Cl2, washed with
water and brine, dried over MgSO4 and concentrated in vacuum,
the residue was purified by column chromatography (10:1 to
5:1, petroleum ether/EtOAc).
TLC: Rf = 0.57 (cyclohexane/EtOAc, 2:1);
[a]D = +28.5 (c = 3.5,
CHCl3); mp = 72–75 °C; 1H NMR (300 MHz, CDCl3): d = 8.03–7.11
(m, 31H, H-Ar), 6.78 (dd, J5,3 = 2.9 Hz, J5,6 = 8.8 Hz, 1H, H-5, dime-
thoxybenzene), 6.68 (d, J6,5 = 8.8 Hz, 1H, H-6, dimethoxybenzene),
6.28 (d, J4 , = 2.9 Hz, 1H, H-40), 5.97 (t, J2 ,3 = J2 ,1 = 9.9 Hz, 1H,
0
0
0
0
0
30
H-20), 5.85 (dd, J3 ,4 = 3.3 Hz, J3 ,2 = 10.3 Hz, 1H, H-30), 5.26 (d,
0
0
0
0
J1 ,2 = 9.9 Hz, 1H, H-10), 4.31 (dd, J6 b,5 = 5.9 Hz, J6 a,5 = 8.1 Hz, 1H,
0
0
0
0
0
H-50), 3.82, 3.68 (2s, 6H, 2 ꢂ OMe), 3.54 (dd, J6 b,5 = 5.5 Hz,
0
0
J6 b,6 a = 8.8 Hz, 1H, H-60b), 3.35 (t, J6 a,6 b = J6 a,5 = 8.4 Hz, 1H, H-
60a); 13C NMR (75 MHz, CDCl3): d = 165.7, 165.2, 164.7 (C@O),
153.6, 151.2 (C-1, C-4, dimethoxybenzene), 143.3, 128.5, 127.7,
133.0, 132.9, 132.7, 129.8, 129.8, 129.5, 129.8, 129.4, 129.3,
128.4, 128.1, 128.1, 126.8 (C-Ar), 126.2, 114.4, 114.0, 111.5 (C-2,
C-3, C-5, C-6, dimethoxybenzene), 86.8, 76.4, 73.5, 71.1, 68.9 (C-
10 to C-50), 61.0 (C-60), 55.8, 55.5 (2 ꢂ OMe). HRESIMS: calcd for
C54H46O10Na: 877.2989; found: m/z 877.2997.
0
0
0
0
0
0
3.1.1.1. 2-(20,30,40-Tri-O-benzoyl-60-O-triphenylmethyl-b-
copyranosyl)-1,4-dimethoxy benzene (2). From compound
D-glu-
1 (402 mg, 0.86 mmol), column chromatography (10:1 to 5:1,
petroleum ether/EtOAc) afforded 2 as a white solid (449 mg,
3.1.2. General procedure for the detritylation
Compounds 3 and 7 were synthesized by this procedure. 6-O-
triphenylmethyl b-C-aryl glycosides were dissolved in CH2Cl2
61%). TLC: Rf = 0.55 (cyclohexane/EtOAc, 2:1);
[a
]D = ꢁ19.3

9760
L. Lin et al. / Bioorg. Med. Chem. 16 (2008) 9757–9763
(5 mL) containing TFA (0.3 mL) and water (0.05 mL). The reaction
mixture was kept for 3–5 h at rt, then diluted with CH2Cl2, washed
with water, saturated aq NaHCO3 and water. The organic phase
was separated, dried and concentrated in vacuum. The residue
was purified by column chromatography.
leum ether/EtOAc) afforded 4 as a white solid (513 mg, 57%).
TLC: Rf = 0.58 (cyclohexane/EtOAc, 3:2); [a]D = +24.2 (c = 0.98,
CHCl3); mp = 131–133 °C; 1H NMR (300 MHz, CDCl3): d = 8.01–
7.29 (m, 15H, 3 ꢂ OCOPhH), 7.19 (d, J3,5 = 2.9 Hz, 1H, H-3, dime-
thoxybenzene), 6.80 (dd, J5,3 = 2.9 Hz, J5,6 = 8.8 Hz, 1H, H-5,
dimethoxybenzene), 6.67 (d, J6,5 = 8.8 Hz, 1H, H-6, dimethoxyben-
3.1.2.1. 2-(20,30,40-Tri-O-benzoyl-b-
D
-glucopyranosyl)-1,4-dime-
zene), 6.07 (t, J3 ,4 = J2 ,3 = 9.6 Hz, 1H, H-30), 5.73 (t, J2 ,1 = J2 ,3
=
0
0
0
0
0
0
0
0
9.5 Hz, 1H, H-20), 5.68 (t, J3 ,4 = J4 ,5 = 9.6 Hz, 1H, H-40), 5.37 (d,
0
0
0
0
thoxybenzene (3). From compound 2 (425 mg, 0.50 mmol), col-
umn chromatography (5:1 to 2:1, petroleum ether/EtOAc)
afforded 3 as a white solid (240 mg, 79%). TLC: Rf = 0.31 (cyclohex-
J1 ,2 = 9.9 Hz, 1H, H-10), 4.17 (m, 1H, H-50), 3.82, 3.65 (2s, 6H,
0
0
2 ꢂ OMe), 3.61 (dd, J6 a,5 = 2.6 Hz, J6 a,6 b = 12.9 Hz, 1H, H-60a), 3.43
0
0
0
0
D = ꢁ52.3 (c = 0.67, CHCl3); mp = 75–78 °C; 1H
(dd, J6 b,5 = 5.5 Hz, J6 a,6 b = 13.2 Hz, 1H, H-60b); 13C NMR (75 MHz,
CDCl3): d = 165.8, 165.2, 164.6 (C@O), 153.8, 151.0 (C-1, C-4, dime-
thoxybenzene), 133.4, 133.0, 132.8, 129.8, 129.6, 129.5, 129.2,
129.0, 128.8, 128.4, 128.1, 128.0 (C-Ar), 125.1, 115.8, 112.3, 111.5
(C-2, C-3, C-5, C-6, dimethoxybenzene), 77.7, 74.5, 73.8, 73.2,
70.3 (C-10 to C-50), 55.7, 55.7 (2 ꢂ OMe), 51.1 (C-60). HRESIMS: calcd
for C35H31N3O9Na: 660.1958; found: m/z 660.1957.
0
0
0
0
ane/EtOAc, 2:1); [a]
NMR (300 MHz, CDCl3): d = 7.99–7.21 (m, 15H, 3 ꢂ OCOPhH),
7.13 (d, J3, 5 = 2.9 Hz, 1H, H-3, dimethoxybenzene), 6.76 (dd,
J5,3 = 2.9 Hz, J5,6 = 8.8 Hz, 1H, H-5, dimethoxybenzene), 6.65 (d,
0
0
0
0
J6,5 = 8 Hz, 1H, H-6, dimethoxybenzene), 6.07 (t, J3 ,2 = J4 ,3 = 9.5
Hz, 1H, H-30), 5.73 (t, J2 ,1 = J2 ,3 = 9.6 Hz, 1H, H-20), 5.61 (t,
0
0
0
0
J3 ,4 = J4 ,5 = 9.5 Hz, 1 H, H-40), 5.30 (d, J1 ,2 = 9.5 Hz, 1H, H-10),
3.96 (m, 1H, H-50), 3.85 (m, 2H, H-60a, H-60b), 3.79, 3.65 (2s, 6H,
2 ꢂ OMe), 2.52 (br s, 1H, OH); 13C NMR (75 MHz, CDCl3):
d = 166.2, 166.1, 164.9 (C@O), 153.9, 151.5 (C-1, C-4, dimethoxy-
benzene), 133.7, 133.2, 133.1, 130.1, 129.9, 129.8, 130.0, 129.4,
129.3, 128.6, 128.4, 128.3 (C-Ar), 125.6, 115.5, 113.4, 111.8 (C-2,
C-3, C-5, C-6, dimethoxybenzene), 79.2, 74.8, 73.9, 73.2, 70.2 (C-
10 to C-50), 62.1 (C-60), 56.1, 55.9 (2 ꢂ OMe). HRESIMS: calcd for
C35H32O10Na: 635.1893; found: m/z 635.1885.
0
0
0
0
0
0
3.1.3.2. 2-(60-Azido-20,30,40-tri-O-benzoyl-60-deoxy-b-
pyranosyl)-1,4-dimethoxy benzene (8). From compound 7
D-galacto-
(286 mg, 0.47 mmol), column chromatography (5:1 to 3:1, petro-
leum ether/EtOAc) afforded 8 as a white solid (231 mg, 78%).
TLC: Rf = 0.50 (cyclohexane/EtOAc, 2:1);
[a]D = +22.2 (c = 4.0,
CHCl3); mp = 62–64 °C; 1H NMR (300 MHz, CDCl3): d = 8.15–7.24
(m, 15H, 3 ꢂ OCOPhH), 7.22 (d, J3,5 = 2.9 Hz, 1H, H-3, dimethoxy-
benzene), 6.77 (dd, J5,3 = 2.9 Hz, J5,6 = 8.8 Hz, 1H, H-5, dimethoxy-
benzene), 6.67 (d, J6,5 = 8.8 Hz, 1H, H-6, dimethoxybenzene), 6.03
3.1.2.2. 2-(20,30,40-Tri-O-benzoyl-b-
thoxybenzene (7). From compound 6 (757 mg, 0.89 mmol),
D-galactopyranosyl)-1,4-dime-
(t, J2 ,3 = J2 ,1 = 9.9 Hz, 1H, H-20), 5.97 (d, J4 ,3 = 3.3 Hz, 1H, H-40),
0
0
0
0
0
0
5.72 (dd, J3 ,4 = 3.3 Hz, J3 ,2 = 10.3 Hz, 1H, H-30), 5.28 (d,
0
0
0
0
column chromatography (5:1 to 2:1, petroleum ether/EtOAc) affor-
J1 ,2 = 10.3 Hz, 1H, H-10), 4.23 (m, 1H, H-50), 3.80, 3.66 (2s, 6H,
0
0
ded 7 as a white solid (302 mg, 56%). TLC: Rf = 0.20 (cyclohexane/
EtOAc, 2:1); [a]
D = +76.5 (c = 0.75, CHCl3); mp = 126–129 °C; 1H
2 ꢂ OMe), 3.61 (dd, J6 a,5 = 7.1 Hz, J6 a,6 b = 12.8 Hz, 1H, H-60a), 3.43
0
0
0
0
(dd, J6 b,5 = 4.8 Hz, J6 a,6 b = 12.9 Hz, 1H, H-60b); 13C NMR (75 MHz,
CDCl3): 165.6, 165.6, 164.8 (C@O), 153.6, 151.3 (C-1, C-4, dime-
thoxybenzene), 133.5, 133.1, 132.8, 129.8, 129.7, 129.5, 129.3,
129.2, 128.9, 128.6, 128.1, 128.0 (C-Ar), 125.8, 114.7, 113.7, 111.5
(C-2, C-3, C-5, C-6, dimethoxybenzene), 76.6, 73.3, 70.6, 69.5 (C-
10 to C-50), 55.8, 55.6 (2 ꢂ OMe), 51.0 (C-60). HRESIMS: calcd for
C35H31N3O9Na: 660.1958; found: m/z 660.1969.
0
0
0
0
NMR (300 MHz, CDCl3): d = 8.18–7.23 (m, 15H, 3 ꢂ OCOPhH),
7.17 (d, J3,5 = 3.3 Hz, H-3, 1H, dimethoxybenzene), 6.77 (dd,
J5,3 = 2.9 Hz, J5,6 = 8.8 Hz, 1H, H-5, dimethoxybenzene), 6.67 (d,
J6,5 = 8.8 Hz,
1H,
H-6,
dimethoxybenzene),
6.10
(t,
J2 ,3 = J2 ,1 = 9.9 Hz, 1H, H-20), 5.93 (d, J4 ,3 = 3.3 Hz, 1H, H-40), 5.74
0
0
0
0
0
0
(dd, J3 ,4 = 3.3 Hz, J3 ,2 = 10.0 Hz, 1H, H-30), 5.26 (d, J1 ,2 = 9.9 Hz,
1H, H-10), 4.19 (m, 1H, H-50), 3.84–3.63 (m, 2H, H-60a, H-60b),
3.78, 3.67 (2s, 6H, 2 ꢂ OMe), 2.59 (br s, 1H, OH); 13C NMR
(75 MHz, CDCl3): d = 166.7, 165.5, 164.9 (C@O), 153.6, 151.3 (C-1,
C-4, dimethoxybenzene), 133.6, 133.1, 132.8, 129.9, 129.7, 129.5,
129.3, 129.0, 129.0, 128.6, 128.2, 128.0 (C-Ar), 126.0, 114.6,
114.0, 111.6 (C-2, C-3, C-5, C-6, dimethoxybenzene), 78.0, 73.4,
70.9, 69.8 (C-10 to C-50), 61.0 (C-60), 55.8, 55.6 (2 ꢂ OMe). HRESIMS:
calcd for C35H32O10Na: 635.1893; found: m/z 635.1901.
0
0
0
0
0
0
3.1.4. General procedure for the 6-O-silylation and 2,3,4-O-
acetylation
Compounds 10 and 14 were synthesized by this procedure. To a
soln of aryl glycosides 933 or 1333 in dry pyridine (2–3 mL),
TBDMSCl (1.5 equiv) and DMAP (0.2 equiv) were added at 0 °C.
After stirring at 0 °C for 30 min, the reaction mixture was warmed
to rt and stirred continuously for 20 h, Ac2O (5 equiv) was added at
rt and the reaction mixture kept stirring until TLC indicated com-
pletion of the reaction. The resulting mixture was diluted with
water and then extracted with CH2Cl2. The organic layer was
washed with water and brine, dried over MgSO4 and concentrated
in vacuum to give a crude residue, and then purified by column
chromatography.
3.1.3. General procedure for 6-azidation
Compounds 4, 8, 12, and 16 were synthesized by this procedure.
To a soln of sugar alcohol in dry CH2Cl2 (3–5 mL), MsCl (1.5 equiv)
and Et3N (1.8 equiv) were added at 0 °C. The ice-bath was removed
and stirring was continued for 18 h. After which, MeOH (around
100 lL) was added, the soln was concentrated. The residue was
dissolved in EtOAc (20–30 mL), washed with H2O, aq NaHCO3
(5%) and brine successively. The organic layer was dried over
MgSO4, filtered and concentrated to an oil, which was used directly
without purification. The mesylate was dissolved in dried DMF (2–
3 mL) and NaN3 (5 equiv) was added. The reaction mixture was
heated to 90 °C for around 20 h. After TLC indicated the formation
of a single product, the mixture was concentrated in vacuum. The
residue was diluted in EtOAc (30 mL), washed successively with
water and brine, dried with MgSO4, filtered and concentrated.
The crude product was purified by column chromatography.
3.1.4.1. 2-(20,30,40-Tri-O-acetyl-60-O-tert-butyldimethylsilyl-b-
glucopyranosyl)-1,4-di methoxynaphthalene (10). From
D-
compound 9 (369 mg, 1.05 mmol), column chromatography (10:1
to 5:1, petroleum ether/EtOAc) afforded 10 as a white solid
(307 mg, 49%). TLC: Rf = 0.47 (petroleum ether/EtOAc, 3:1);
[a]
D = –3.05 (c = 0.3, CHCl3); mp = 46–48 °C; 1H NMR (250 MHz,
CDCl3): d = 8.22, 8.03 (2d, J = 7.5 Hz, 2H, H-5, H-8, ArH), 7.51 (m,
2H, H-6, H-7, ArH), 6.77 (s, 1H, H-3, ArH), 5.42 (m, 2H, H-20, H-
30), 5.32 (m, 1H, H-40), 5.07 (d, J1 ,2 = 10.0 Hz, 1H, H-10), 3.97, 3.92
(2s, 6H, 2 ꢂ OMe), 3.80–3.73 (m, 3H, H-50, H-60a, H-60b), 2.06,
2.02, 1.71 (3s, 9H, 3 ꢂ OCOCH3), 0.83 (s, 9H, tBu), ꢁ0.06, ꢁ0.01
(2s, 6H, 2 ꢂ Me); HRESIMS: calcd for C30H42O10SiNa: 613.2445;
found: m/z 613.2432.
0
0
3.1.3.1. 2-(60-Azido-20,30,40-tri-O-benzoyl-60-deoxy-b-
anosyl)-1,4-dimethoxy benzene (4). From compound
(872 mg, 1.42 mmol), column chromatography (6:1 to 5:1, petro-
D-glucopyr-
3

L. Lin et al. / Bioorg. Med. Chem. 16 (2008) 9757–9763
9761
3.1.4.2. 2-(20,30,40-Tri-O-acetyl-60-O-tert-butyldimethylsilyl-b-
D
-
matography (3:1, petroleum ether/EtOAc) afforded 12 as a white
solid (251 mg, 83%). TLC: Rf = 0.69 (cyclohexane/EtOAc, 1:1);
galactopyranosyl)-1,4-dimethoxynaphthalene (14).
From
compound 1333 (101.8 mg, 0.29 mmol), column chromatography
(10:1 to 5:1, petroleum ether/EtOAc) afforded 14 as a white solid
(112 mg, 65%). TLC: Rf = 0.60 (cyclohexane/EtOAc, 2:1);
[a]
D = +5.2 (c = 1.6, CHCl3); mp = 143–145 °C; 1H NMR (300 MHz,
CDCl3): d = 8.22, 8.03 (2d, J = 7.7 Hz, 2H, H-5, H-8, ArH), 7.51 (m,
2 H, H-6, H-7, ArH), 6.78 (s, 1H, H-3, ArH), 5.45 (m, 2H, H-20, H-
D = ꢁ11.7 (c = 1.4, CHCl3); mp = 53–56 °C; 1H NMR (300 MHz,
40), 5.34 (t, J3 4 = J3 ,2 = 9.6 Hz, 1H, H-30), 5.14 (d, J1 ,2 = 9.2 Hz, 1H,
H-10), 3.97, 3.93 (2s, 6H, 2 ꢂ OMe), 3.95 (m, 1H, H-50), 3.52 (dd,
0
0
0
0
0
0
[a]
CDCl3): d = 8.22, 8.05 (2d, J = 7.5 Hz, 2H, H-5, H-8, ArH), 7.52 (m,
2H, H-6, H-7, ArH), 6.82 (s, 1H, H-3, ArH), 5.70 (m, 2H, H-20, H-
J6 a,5 = 2.2 Hz, J6 a,6 b = 13.6 Hz, 1H, H-60a), 3.21 (dd, J6 b,5 = 4.8 Hz,
0
0
0
0
0
0
40), 5.31 (dd, J3 ,4 = 3.3 Hz, J3 ,2 = 9.9 Hz, 1H, H-30), 5.08 (d,
J6 a,6 b = 13.6 Hz, 1H, H-60b), 2.06, 2.02, 1.71 (3s, 9H, 3 ꢂ OCOCH3);
13C NMR (75 MHz, CDCl3): d = 170.2, 169.4, 168.8 (C@O), 152.3,
148.3 (C-1, C-4, ArC), 127.9, 127.0, 126.7, 126.0, 123.4, 122.5,
122.1 (C-2, C-5 to C-10, ArC), 101.1 (C-3, ArC), 76.6, 74.5, 73.9,
71.2, 69.5 (C-10 to C-50), 63.1, 55.6 (2 ꢂ OMe), 50.7 (C-60), 20.5,
20.3 (3 ꢂ OCOCH3). HRESIMS: calcd for C24H27N3O9Na: 524.1645;
found: m/z 524.1651.
0
0
0
0
0
0
J1 ,2 = 9.9 Hz, 1H, H-10), 4.01, 3.93 (2s, 6H, 2 ꢂ OMe), 3.98 (m, 1H,
H-50), 3.76–3.59 (m, 2H, H-60a, H-60b), 2.23, 2.00, 1.72 (3s, 9H,
3 ꢂ OCOCH3), 0.84 (s, 9H, tBu), ꢁ0.01, ꢁ0.01 (2s, 6H, 2 ꢂ Me); 13C
NMR (75 MHz, CDCl3): d = 170.4, 170.2, 169.4 (C@O), 152.3, 148.9
(C-1, C-4, ArC), 128.3, 127.2, 126.8, 126.2, 124.1, 122.6, 122.4 (C-
2, C-5 to C-10, ArC), 102.1 (C-3, ArC), 78.0, 74.9, 73.1, 68.8, 68.1
(C-10 to C-50), 63.5 (OCH3), 60.9 (C-60), 55.8 (OMe), 25.8 (tBu),
21.0, 20.8, 20.7 (3 ꢂ OCOCH3), 18.3 (Cq, tBu), ꢁ5.4, ꢁ5.6 (2 ꢂ Me).
HRESIMS: calcd for C30H42O10SiNa: 613.2445; found: m/z
613.2437.
0
0
3.1.7. 2-(20,30,40-Tri-O-acetyl-60-azido-60-deoxy-b-
D-
galactopyranosyl)-1,4-dimethoxy naphthalene (16)
Compound 15 (91 mg, 0.19 mmol) was treated with MsCl/Et3N
and NaN3 according to the general procedure. Column chromatog-
raphy (4:1, petroleum ether/EtOAc) afforded 16 as a pale yellow
3.1.5. General procedure for the deprotection of silyl group
Compounds 11 and 15 were synthesized by this procedure. To a
stirred soln of silylated glycosides (10 or 14) in dry MeOH (2–3 mL)
was added AcCl (0.15 equiv) at ice-bath temperature. The reaction
mixture was stirred at rt for 5 h. After completion of the reaction
(monitored by TLC), CH2Cl2 (15–20 mL) was added, the reaction
mixture was neutralized with 5% NaHCO3 (2–3 mL) and washed
with H2O (5 mL). Finally, the organic layer was dried over MgSO4
and concentrated in vacuum to give a crude residue, which was
used directly in next step or purified by column chromatography.
oil (64 mg, 67%). TLC: Rf = 0.42 (cyclohexane/EtOAc, 2:1); [a] =
D
–0.7 (c = 0.29, CHCl3); 1H NMR (300 MHz, CDCl3): d = 8.23, 8.05
(2d, J = 7.4 Hz, 2H, H-5, H-8, ArH), 7.55 (m, 2H, H-6, H-7, ArH),
6.82 (s, 1H, H-3, ArH), 5.71 (t, J2 ,3 = J2 ,1 = 9.9 Hz, 1H, H-20), 5.56
0
0
0
0
(d, J4 ,3 = 2.9 Hz, 1H, H-40), 5.28 (dd, J3 ,4 = 3.3 Hz, J3 ,2 = 10.3 Hz,
0
0
0
0
0
0
1H, H-30), 5.10 (d, J1 ,2 = 10.3 Hz, 1H, H-10), 4.07 (m, 1H, H-50),
0
0
0
0
4.02, 3.95 (2s, 6H, 2 ꢂ OMe), 3.50 (d, J6 a,5 = 7.0 Hz,
0
0
0
0
J6 a,6 b = 12.9 Hz,
1H,
H-60a),
3.32
(dd,
J6 b,5 = 5.2 Hz,
J6 a,6 b = 12.9 Hz, 1H, H-60b), 2.28, 2.01, 1.72 (3s, 9H, 3 ꢂ OCOCH3);
13C NMR (75 MHz, CDCl3): d = 170.5, 170.3, 169.3 (C@O), 152.4,
148.9 (C-1, C-4, ArC), 128.2, 127.2, 126.9, 126.3, 123.7, 122.7,
122.4 (C-2, C-5 to C-10, ArC), 101.9 (C-3, ArC), 76.4, 72.7, 68.9,
68.3 (C-10 to C-50), 63.4, 55.8 (2 ꢂ OMe), 50.9 (C-60), 21.0, 20.8,
20.6 (3 ꢂ OCOCH3). HRESIMS: calcd for C24H27N3O9Na: 524.1645;
found: m/z 524.1640.
0
0
3.1.5.1. 2-(20,30,40-Tri-O-acetyl-b-
thoxynaphthalene (11). From compound 10 (452 mg,
D-glucopyranosyl)-1,4-dime-
0.77 mmol), the crude product 11 was obtained as a colorless oil
(360 mg, 99%). TLC: Rf = 0.19 (cyclohexane/EtOAc, 3:2); 1H NMR
(250 MHz, CDCl3): d = 8.23, 8.04 (2d, J = 7.5 Hz, 2H, H-5, H-8,
ArH), 7.52 (m, 2H, H-6, H-7, ArH), 6.75 (s, 1H, H-3, ArH), 5.52–
5.47 (m, 2H, H-30, H-40), 5.24 (t, J2 ,1 = J2 ,3 = 9.5 Hz, 1H, H-20),
3.1.8. Dipropargyl isophthalate (17)
To a suspension of isophthaloyl chloride (693 mg, 3.41 mmol)
and DMAP (83 mg, 0.68 mmol) in dry CH2Cl2 (15 mL), propargyl
0
0
0
0
5.14 (d, J1 ,2 = 9.5 Hz, 1H, H-10), 4.00, 3.93 (2s, 6H, 2 ꢂ OCH3),
3.81–3.65 (m, 3H, H-50, H-60a, H-60b), 2.10, 2.03, 1.72 (3s, 9H,
3 ꢂ OCOCH3). HRESIMS: calcd for C24H28O10Na: 499.1580; found:
m/z 499.1573.
0
0
alcohol (403 lL, 6.83 mmol), and Et3N (3.2 mL, 22.5 mmol) were
added at 0 °C. The mixture was stirred at 0 °C for 3 h and then
warmed to rt for another 48 h, filtered, and the precipitate was
rinsed with CH2Cl2. The combined filtrate was washed with 1 M
HCl and brine, dried over MgSO4 and concentrated in vacuum to
give a pale yellow powder. The crude product was purified by col-
umn chromatography (2:1, petroleum ether/EtOAc) to afford 17 as
a white powder (598 mg, 72%). TLC: Rf = 0.64 (cyclohexane/EtOAc,
2:1); mp = 82–84 °C; 1H NMR (300 MHz, CDCl3): d = 8.74 (s, 1H, H-
2, ArH), 8.28 (dd, 2H, J = 1.9 and 7.7 Hz, H-4, H-6, ArH), 7.57 (t, 1H,
J = 7.7 Hz, H-5, ArH), 4.96 (d, 4H, J = 1.8 Hz, 2 ꢂ CH2), 2.54 (t, 2H,
J = 1.8 Hz, 2 ꢂ CH); 13C NMR (100 MHz, CDCl3): d = 164.8 (C@O),
134.4, 131.1, 128.8 (Ar), 130.0 (Cq, Ar), 77.4 (Cq), 75.3 (CH), 52.8
(CH2).
3.1.5.2. 2-(20,30,40-Tri-O-acetyl-b-
D-galactopyranosyl)-1,4-dime-
thoxynaphthalene (15). From compound 14 (352 mg, 0.60
mmol), column chromatography (5:1 to 2:1, petroleum ether/
EtOAc) afforded 15 as a pale brown solid (107 mg, 38%). TLC:
Rf = 0.29 (cyclohexane/EtOAc, 2:1); [a]D = +6.0 (c = 0.84, CHCl3);
mp = 67–70 °C; 1H NMR (300 MHz, CDCl3): d = 8.23, 8.04 (2d,
J = 7.7 Hz, 2H, H-5, H-8, ArH), 7.53 (m, 2H, H-6, H-7, ArH), 6.80 (s,
1H, H-3, ArH), 5.75 (t, J2 ,1 = J2 ,3 = 9.9 Hz, 1H, H-20), 5.56 (d,
0
0
0
0
J4 ,3 = 3.3 Hz, 1H, H-40), 5.32 (dd, J3 ,4 = 3.7 Hz, J3 ,2 = 9.9 Hz, 1H,
0
0
0
0
0
0
H-30), 5.10 (d, J1 ,2 = 9.9 Hz, 1H, H-10), 4.04 (m, 1H, H-50), 4.02,
0
0
0
0
0
0
3.93 (2s, 6H, 2 ꢂ OMe), 3.74 (dd, J6 a,5 = 7.0 Hz, J6 a,6 b = 11.8 Hz,
1H, H-60a), 3.93 (dd, J6 b,5 = 5.9 Hz, J6 b,6 a = 11.8 Hz, 1H, H-60b),
2.27, 2.02, 1.73 (3s, 9H, 3 ꢂ OCOCH3); 13C NMR (75 MHz, CDCl3):
d = 171.3, 170.3, 169.4 (C@O), 152.4, 149.0 (C-1, C-4, ArC), 128.3,
127.2, 127.0, 126.3, 123.9, 122.7, 122.4 (C-2, C-5 to C-10, ArC),
102.0 (C-3, ArC), 78.2, 75.0, 72.8, 69.0, 68.7 (C-10 to C-50), 63.5
(OMe), 61.2 (C-60), 55.9 (OMe), 21.0, 20.8, 20.7 (3 ꢂ OCOCH3). HRE-
SIMS: calcd for C24H28O10Na: 499.1580; found: m/z 499.1562.
0
0
0
0
3.1.9. General procedure for the Huisgen cycloaddition
Compounds 18, 19, 22, and 23 were synthesized by this proce-
dure. 6-Azido C-aryl glycoside (2 equiv) was dissolved in CH2Cl2
(1.5 mL), dipropargyl ester 17 (1 equiv), H2O (1.5 mL), copper sul-
fate (0.2 equiv) and sodium ascorbate (0.4 equiv) were added suc-
cessively. The resulting mixture was stirred at rt until TLC
indicated the disappearance of the starting materials. The mixture
was diluted with CH2Cl2 (15 mL), washed with water, dried over
MgSO4, filtered and concentrated to give the desired compound.
If the residue was chromatographically not uniform, it was purified
by column chromatography.
3.1.6. 2-(20,30,40-Tri-O-acetyl-60-azido-60-deoxy-b-
glucopyranosyl)-1,4-dimethoxy naphthalene (12)
D-
Compound 11 (307 mg, 0.64 mmol) was treated with MsCl/
Et3N and NaN3 according to the general procedure. Column chro-

9762
L. Lin et al. / Bioorg. Med. Chem. 16 (2008) 9757–9763
3.1.9.1. Dimeric aryl-b-C-
pound 4 (108 mg, 0.17 mmol), Huisgen cycloaddition afforded 18
as a pink solid without purification (121 mg, 94%). TLC: Rf = 0.19
D
-glucopyranoside 18.
From com-
70.6, 70.0 (C-10 to C-50), 63.1 (OCH3), 58.1 (CH2), 55.6 (OMe), 51.0
(C-60), 20.6, 20.5, 20.2 (OCOCH3); HRESIMS: calcd for C62H64N6O22Na:
1267.3971; found: m/z 1267.3981.
(cyclohexane/EtOAc, 1:1);
[a]D = –18.2 (c = 0.68, CHCl3);
mp = 133–135 °C; 1H NMR (300 MHz, CDCl3): d = 8.63 (s, 1H, H-
200, Phth), 8.17 (d, J = 7.7 Hz, 2H, H-400, H-600 Phth), 8.02–7.25 (m,
33H, Ar-H, 2 ꢂ CH triazole), 7.12 (d, J3,5 = 2.6 Hz, 2H, 2 ꢂ H-3,
dimethoxybenzene), 6.76 (dd, J5,3 = 2.9 Hz, J5,6 = 8.8 Hz, 2H,
2 ꢂ H-5, dimethoxybenzene), 6.63 (d, J6,5 = 8.8 Hz, 2H, 2 ꢂ H-6,
3.1.9.4. Dimeric aryl-b-C-D-galactopyranoside 23.
From com-
pound 16 (49 mg, 0.098 mmol), Huisgen cycloaddition afforded 23 as
a pale yellow solid without purification (58 mg, 95%). TLC: Rf = 0.11
(cyclohexane/EtOAc, 1:1); [a]D = +8.2 (c = 3.2, CHCl3); mp = 88–92 °C;
1H NMR (300 MHz, CDCl3): d = 8.48 (s, 1H, H-200, Phth), 8.20, 7.90
(2m, 4H, 2 ꢂ H-5, 2 ꢂ H-8, dimethoxynaphthalene), 8.00 (d,
J = 7.4 Hz, 2H, H-400, H-600, Phth), 7.49 (m, 6 H, 2 ꢂ CH triazole,
2 ꢂ H-6, 2 ꢂ H-7 dimethoxynaphthalene), 7.25 (t, J = 7.7 Hz, 1H,
H-400, Phth), 6.78 (s, 2H, 2 ꢂ H-3, dimethoxynaphthalene), 5.74 (t,
dimethoxybenzene), 6.08 (t, J3 ,2 = J3 ,4 = 9.3 Hz, 2H, 2 ꢂ H-30),
0
0
0
0
5.81
(t,
0
J2 ,1 = J2 ,3 = 9.5 Hz,
2H,
2 ꢂ H-20),
5.59
(t,
0
0
0
0
J4 ,5 = J4 ,3 = 9.5 Hz, 2H, 2 ꢂ H-40), 5.44 (s, 4H, 2 ꢂ CH2), 5.26 (d,
0
0
0
J1 ,2 = 9.9 Hz, 2H, 2 ꢂ H-10), 4.85 (d, J6 a,6 b = 14.3 Hz, 2H, 2 ꢂ H-
0
0
0
0
60a), 4.59 (dd, J6 b,5 = 8.1 Hz, J6 b,6 a = 14.4 Hz, 2H, 2 ꢂ H-60b), 4.43
(m, 2H, 2 ꢂ H-50), 3.78, 3.59 (2s, 12H, 4 ꢂ OMe); 13C NMR
(75 MHz, CDCl3): d = 165.7, 165.5, 165.2, 164.6 (C@O), 153.6,
151.2 (C-1, 2 ꢂ C-4, dimethoxybenzene), 134.0, 133.6, 133.1,
133.0, 130.9, 130.1, 129.9, 129.6, 129.5, 129.0, 128.8, 128.5,
128.4, 128.2, 128.0, 125.5 (C-Ar, C-triazole), 124.6, 115.8, 112.3,
111.5 (C-2, C-3, C-5, C-6, dimethoxybenzene), 77.0 (C-50), 74.3
(C-30), 74.0 (C-10), 72.3 (C-20), 70.8 (C-40), 58.2 (CH2), 55.7, 55.6
(OMe), 51.6 (C-60); HRESIMS: calcd for C84H72N6O22Na:
1539.4597; found: m/z 1539.4618.
J2 ,3 = J2 ,1 = 9.9 Hz, 2H, H-20), 5.61 (d, J4 ,3 = 2.9 Hz, 2H, H-40), 5.35 (s,
0
0
0
0
0
0
0
0
0
0
4H, 2 ꢂ CH2), 5.29 (dd, J3 ,4 = 2.9 Hz, J3 ,2 = 9.9 Hz, 2H, 2 ꢂ H-30), 5.04
0
0
0
0
(d, J1 ,2 = 10.3 Hz, 2H, 2 ꢂ H-10), 4.64–4.45 (m, 6H, 2 ꢂ H-50, 2 ꢂ H-60a,
2 ꢂ H-60b), 4.01, 3.61 (2s, 12H, 4 ꢂ OMe), 2.28, 2.00, 1.70 (3s, 18H,
6 ꢂ COCH3); 13C NMR (75 MHz, CDCl3): d = 170.3, 170.1, 169.3, 165.2
(C@O), 152.2, 149.1 (C-1, C-4, dimethoxynaphthalene), 142.9 (Cq, tria-
zole), 134.0, 130.8, 130.1, 127.9 (C-100 to C-600, Phth), 128.1, 127.1,
126.9, 126.3, 123.2, 122.6, 122.3 (C-2, C-5 to C-10, dimethoxynaphtha-
lene), 125.3 (CH triazole), 101.8 (C-3, dimethoxynaphthalene), 76.1,
74.9, 72.5, 68.9, 67.9 (C-10 to C-50), 63.3 (OCH3), 58.9 (CH2), 55.8
(OMe), 49.1 (C-60), 20.9, 20.7, 20.6 (OCOCH3); HRESIMS: calcd for
C62H64N6O22Na: 1267.3971; found: m/z 1267.3971.
0
0
3.1.9.2. Dimeric aryl-b-C-D-galactopyranoside 19. From com-
pound 8 (51 mg, 0.08 mmol), column chromatography (1:1, petro-
leum ether/EtOAc) afforded 19 as a pale yellow solid (54 mg, 90%).
3.1.10. General procedure for oxidation of 1,4-dimethoxy
benzene or naphthalene derivatives
TLC: Rf = 0.17 (cyclohexane/EtOAc, 1:1); [a]D = +130.7 (c = 0.41,
CHCl3); mp = 110–113 °C; 1H NMR (300 MHz, CDCl3): d = 8.58 (s,
1H, H-2000 Phth), 8.23–7.19 (m, 35H, Ar-H, 2 ꢂ CH triazole), 7.12 (d,
Compounds 18, 19, 22, and 23 were oxidized by this procedure.
A soln of CAN (3 equiv) in water (1–2 mL/mmol) was added to a
soln of the 1,4-dimethoxy benzene or naphthalene derivative in
MeCN (1–2 mL/mmol). After stirring at rt in the dark for 1 h, the
mixture was extracted with CH2Cl2 (3 ꢂ 15 mL). The combined
CH2Cl2 soln was washed with brine and dried over MgSO4, then
concentrated under diminished pressure. The crude product was
purified by column chromatography.
J3,5 = 2.9 Hz, 2H, 2 ꢂ H-3, dimethoxybenzene), 6.75 (dd, J5,3
=
2.9 Hz, J5,6 = 8.8 Hz, 2H, 2 ꢂ H-5, dimethoxybenzene), 6.63 (d,
J6,5 = 8.8 Hz,
2H,
2 ꢂ H-6,
dimethoxybenzene),
6.10
(t,
J2 ,3 = J2 ,1 = 9.9 Hz, 2H, 2 ꢂ H-20), 6.02 (d, J4 ,3 = 3.3 Hz, 2H, 2 ꢂ H-
0
0
0
0
0
0
40), 5.72 (dd, J3 ,4 = 3.3 Hz, J3 ,2 = 10.3 Hz, 2H, 2 ꢂ H-30), 5.40 (s, 4H,
0
0
0
0
2 ꢂ CH2), 5.20 (d, J1 ,2 = 9.9 Hz, 2H, 2 ꢂ H-10), 4.76 (d,
0
0
J6 a,6 b = 11.4 Hz, 2H, 2 ꢂ H-60a), 4.55 (m, 4H, 2 ꢂ H-50, 2 ꢂ H-60b),
3.78, 3.59 (2s, 12H, 4 ꢂ OMe); 13C NMR (75 MHz, CDCl3):
d = 165.6, 165.4, 165.3, 164.8 (C@O), 153.6, 151.4 (C-1, C-4, dime-
thoxybenzene), 142.4 (Cq, triazole), 134.0, 133.8, 133.1, 133.0,
130.9, 130.1, 129.9, 129.6, 129.5, 129.1, 128.9, 128.8, 128.7, 128.2,
128.1, 128.5 (C-Ar), 125.3, 114.6, 113.8, 111.6 (CH triazole, C-2, C-
3, C-5, C-6, dimethoxybenzene), 76.2, 74.4, 73.1, 70.0, 69.6 (C-10 to
C-50), 58.2 (CH2), 55.7, 55.6 (OMe), 51.0 (C-60); HRESIMS: calcd for
C84H72N6O22Na: 1539.4597; found: m/z 1539.4562.
0
0
3.1.10.1. Dimeric b-C-
D-glucopyranosyl 1,4-benzoquinone
20. Oxidation of compound 18 (107 mg, 0.071 mmol) and
purification by column chromatography (1:1 petroleum ether/
EtOAc to 10:1 CH2Cl2/MeOH) afforded 20 as a yellow solid
(98 mg, 95%). TLC: Rf = 0.43 (cyclohexane/EtOAc = 2:3);
[a] =
D
–61.5 (c = 1.2, CHCl3); 1H NMR (300 MHz, CDCl3): d = 8.63 (s, 1H,
Ar-H), 8.22, 8.20 (2s, 2H, CH triazole), 8.00–7.23 (m, 33H, Ar-H),
7.01 (d, 2H, J3,5 = 2.2 Hz, 2 ꢂ H-3 benzoquinone), 6.71 (dd, 2H,
J5,3 = 2.6 Hz, J5,6 = 10.3 Hz, 2 ꢂ H-5 benzoquinone), 6.64 (d, 2H,
0
0
3.1.9.3. Dimeric aryl-b-C-D-glucopyranoside 22.
From com-
J6,5 = 10.3 Hz, 2 ꢂ H-6 benzoquinone), 6.08 (t, 2H, J3 ,4 = 9.6 Hz,
J3 ,2 = 9.2 Hz, 2 ꢂ H-30), 5.58 (m, 4H, 2 ꢂ H-20, 2 ꢂ H-40), 5.48 (s,
0 0
pound 12 (60 mg, 0.12 mmol), column chromatography (1:1 petro-
leum ether/EtOAc to EtOAc) afforded 22 as a pale yellow solid
4H, 2 ꢂ CH2), 4.93 (d, 2H, J1 , = 9.5 Hz, 2 ꢂ H-10), 4.83 (d, 2H,
0
20
J6 a,6 b = 12.9 Hz, 2 ꢂ H-60a), 4.61 (dd, 2H, J6 b,5 = 8.5 Hz,
0
0
0
0
(63 mg, 86%). TLC: Rf = 0.08 (cyclohexane/EtOAc, 1:1); [
a
]
D = ꢁ10.7
(c = 0.36, CHCl3); mp = 138–140 °C; 1H NMR (300 MHz, CDCl3):
d = 8.44 (s, 1H, H-200 Phth), 8.19, 7.91 (2 m, 4H, 2 ꢂ H-5, 2 ꢂ H-8,
dimethoxynaphthalene), 7.95 (dd, J = 1.9 Hz and 7.7 Hz, 2H, H-400,
H-600 Phth), 7.66 (s, 2H, 2 ꢂ CH, triazole), 7.48 (m, 4H, 2 ꢂ H-6,
2 ꢂ H-7, dimethoxynaphthalene), 7.20 (t, J = 7.7 Hz, 1H, H-400 Phth),
6.68 (s, 2H, 2 ꢂ H-3, dimethoxynaphthalene), 5.46 (m, 4H, 2 ꢂ H-
20, 2 ꢂ H-30), 5.32 (s, 4H, 2 ꢂ CH2), 5.06 (m, 4H, 2 ꢂ H-10, 2 ꢂ H-40),
J6 b,6 a = 14.7 Hz, 2 ꢂ H-60b), 4.42 (m, 2H, 2 ꢂ H-50); 13C NMR
(75 MHz, CDCl3): d = 186.6, 184.9, 165.5, 165.5, 165.3, 165.2
(C@O), 143.3 (C-2, benzoquinone), 142.8 (Cq, triazole), 136.3,
136.3 (C-5, C-6, benzoquinone), 134.0, 133.8, 133.5, 133.5, 133.2,
131.0, 130.1, 129.9, 129.7, 129.5, 128.6, 128.5, 128.4, 128.2, 128.1
(Ar), 125.5 (CH triazole), 77.2, 73.4, 72.9, 72.7, 70.3 (C-10 to C-50),
0
0
58.3 (CH2), 51.2 (C-60). HRESIMS: calcd for C80H60N6O22
1479.3658; found: m/z 1479.3660.
:
4.65 (dd, J6 a,5 = 2.6 Hz, J6 a, 6 b = 14.7 Hz, 2H, 2 ꢂ H-60a), 4.42 (dd,
0
0
0
0
J6 b,5 = 7.7 Hz, J6 b,6 a = 14.7 Hz, 2H, 2 ꢂ H-60b), 4.17 (m, 2H, 2 ꢂ H-
50), 3.96, 3.64 (2s, 12H, 4 ꢂ OMe), 2.12, 2.00, 1.68 (3s, 18H,
6 ꢂ OCOCH3); 13C NMR (75 MHz, CDCl3): d = 170.0, 169.7, 168.9,
165.1 (C@O), 152.2, 148.8 (C-1, C-4, dimethoxynaphthalene), 142.5
(Cq, triazole), 133.8, 130.6, 129.9, 128.4 (C-100 to C-600, Phth), 127.9,
127.0, 126.8, 126.2 (C-6, C-7, C-9, C-10, dimethoxynaphthalene),
125.4 (CH, triazole), 122.7, 122.4, 122.1 (C-2, C-5, C-8, dimethoxy-
naphthalene), 101.0 (C-3, dimethoxynaphthalene), 76.6, 74.2, 74.0,
0
0
0
0
3.1.10.2. Dimeric
21. Oxidation of compound 19 (42 mg, 0.028 mmol) afforded
21 as a yellow solid (38 mg, 94%). TLC: Rf = 0.53 (cyclohexane/
EtOAc = 1:2); [
D = +27.9 (c = 1.85, CHCl3); 1H NMR (300 MHz,
b-C-D-galactopyranosyl 1,4-benzoquinone
a]
CDCl3): d = 8.54 (s, 1H, Ar-H), 8.15–7.20 (m, 35H, Ar-H, 2 ꢂ CH tri-
azole), 7.06 (d, 2H, J3,5 = 2.6 Hz, 2 ꢂ H-3, benzoquinone), 6.70 (dd,
2H, J5,3 = 2.6 Hz, J5,6 = 10.3 Hz, 2 ꢂ H-5, benzoquinone), 6.62 (d,

L. Lin et al. / Bioorg. Med. Chem. 16 (2008) 9757–9763
9763
0
0
2H, J6,5 = 10.3 Hz, 2 ꢂ H-6, benzoquinone), 6.04 (t, 2H, J4 ,3 = 2.2 Hz,
plate reader at 405 nm for 2 min at 30 °C and the initial rate of
the hydrolysis was determined using the early linear region of
the enzymatic reaction kinetic curve. For calculating IC50, inhibi-
tion assays were performed with 30 nM recombinant enzyme,
2 mM pNPP in 50 mM MOPS at pH 6.5, and the inhibitors diluted
around the estimated IC50 values. IC50 was calculated from the
nonlinear curve fitting of percent inhibition (inhibition (%)) versus
inhibitor concentration [I] by using the following equation inhibi-
tion (%) = 100/{1 + (IC50/[I])k}, where k is the Hill coefficient.
2 ꢂ H-40), 5.79 (m, 4H, 2 ꢂ H-20, 2 ꢂ H-30), 5.42 (s, 4H, 2 ꢂ CH2),
4.92 (d, 2H, J1 ,2 = 8.5 Hz, 2 ꢂ H-10), 4.73 (d, 2H, J6 a,6 b = 11.4 Hz,
2 ꢂ H-60a), 4.62–4.51 (m, 4H, 2 ꢂ H-60b, 2 ꢂ H-50); 13C NMR
(75 MHz, CDCl3): d = 187.1, 185.1, 165.7, 165.6, 165.4, 165.4
(C@O), 144.0 (C-2, benzoquinone), 142.8 (Cq, triazole), 136.5,
136.4 (C-5, C-6, benzoquinone), 134.2, 134.1, 133.8, 133.7, 133.4,
131.0, 130.2, 130.1, 129.8, 129.8, 129.0, 128.7, 128.6, 128.5, 128.4
(Ar) 125.3 (CH, triazole), 76.5, 73.2, 72.4, 70.6, 69.2 (C-10 to C-50),
0
0
0
0
58.4 (CH2), 50.9 (C-60). HRESIMS: calcd for C80H60N6O22
1479.3658; found: m/z 1479.3673.
:
Acknowledgments
3.1.10.3. Dimeric b-C-
24. Oxidation of compound 22 (51 mg, 0.041 mmol) afforded
24 as a yellow solid (43 mg, 90%). TLC: Rf = 0.18 (cyclohex-
D
-glucopyranosyl 1,4-naphthoquinone
L.L. thanks the Ecole Normale Supérieure de Cachan for a Doc-
torate fellowship. This work was supported by CNRS, ENS Cachan,
National Natural Science Foundation of China (Grant No.
20876045) and Shanghai Science and Technology Community
(No. 074107018).
ane:EtOAc = 1:2); Rf = 0.91 (EtOAc); [a]D = +21.3 (c = 0.52, CHCl3);
1H NMR (300 MHz, CDCl3): d = 8.40 (s, 1H, Ar-H), 8.01–7.88 (m,
6H, Ar-H), 7.79 (s, 2H, 2 ꢂ CH, triazole,), 7.72 (m, 4H, 2 ꢂ H-6,
2 ꢂ H-7, naphthoquinone), 7.22 (t, 1H, J = 7.7 Hz, H-4000 isophtha-
loyl), 7.04 (s, 2H, 2 ꢂ H-3, naphthoquinone), 5.41 (dd, 2H,
References and notes
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R. J. Biol. Chem. 2000, 275, 9792.
0
0
0
0
J3 ,2 = 9.6 Hz, J3 ,4 = 9.2 Hz, 2 ꢂ H-3), 5.39 (s, 4H, 2 ꢂ CH2), 5.06 (t,
2H, J2 ,3 = J2 ,1 = 9.6 Hz, 2 ꢂ H-20), 5.02 (dd, 2H, J4 ,3 = 9.2 Hz,
0
0
0
0
0
0
J4 ,5 = 9.9 Hz, 2 ꢂ H-40), 4.84 (d, 2H, J1 ,2 = 9.5 Hz, 2 ꢂ H-10), 4.69
3. Cheng, A.; Dubé, N.; Gu, F.; Tremblay, M. L. Eur. J. Biochem. 2002, 269, 1050.
4. Elchebly, M.; Payette, P.; Michaliszyn, E.; Cromlish, W.; Collins, S.; Loy, A. L.;
Normandin, D.; Cheng, A.; Himms-Hagen, J.; Chan, C. C.; Ramachandran, C.;
Gresser, M. J.; Tremblay, M. L.; Kennedy, B . P. Science 1999, 283, 1544.
5. Klaman, L. D.; Boss, O.; Peroni, O. D.; Kim, J. K.; Martino, J. L.; Zabolotny, J. M.;
Moghal, N.; Lubkin, M.; Kim, Y. B.; Sharpe, A. H.; Stricker-Krongrad, A.;
Schulman, G. I.; Neel, B. G.; Kahn, B. B. Mol. Cell. Biol. 2000, 20, 5479.
6. Moller, D. E. Nature 2001, 414, 821.
7. Montalibet, J.; Kennedy, B. P. Drug Discov. Today: Ther. Strategies 2005, 2, 129.
8. Zhang, Z. Y. Annu. Rev. Pharmacol. Toxicol. 2002, 42, 209.
9. Bialy, L.; Waldmann, H. Angew. Chem. Int. Ed. 2005, 44, 3814.
10. Dewang, P. M.; Hsu, N.-M.; Peng, S.-Z.; Li, W.-R. Curr. Med. Chem. 2005, 12, 1.
11. Lee, S.; Wang, Q. Med. Res. Rev. 2006, 1.
0
0
0
0
(dd, 2H, J6 a,5 = 2.6 Hz, J6 a,6 b = 14.7 Hz, 2 ꢂ H-60a), 4.44 (dd, 2H,
0
0
0
0
J6 b,5 = 8.5 Hz, J6 a,6 b = 14.7 Hz, 2 ꢂ H-60b), 4.13 (m, 2H, 2 ꢂ H-50),
2.10, 1.98, 1.84 (3s, 18H, 6 ꢂ COCH3); 13C NMR (75 MHz, CDCl3):
d = 184.1, 183.0, 169.8, 169.6, 169.5, 165.1 (C@O), 145.3 (C-2,
naphthaquinone), 142.9 (Cq, triazole), 135.7, 134.1, 134.0 (C-3, C-
5, C-8, naphthaquinone), 133.8, 131.5, 131.5, 130.6, 129.9, 128.4,
126.4, 126.2 (Ar), 125.4 (CH triazole), 76.6, 73.2, 72.5, 72.2, 69.7
(C-10 to C-50), 58.2 (CH2), 51.0 (C-60), 20.6, 20.4, 20.3 (COCH3). HRE-
SIMS: calcd for C58H52N6O22: 1207.3032; found: m/z 1207.3007.
0
0
0
0
12. Zhang, S.; Zhang, Z.-Y. Drug Discov. Today 2007, 12, 373.
13. Boustelis, I. G.; Yu, X.; Zhang, Z.-Y.; Borch, R. F. J. Med. Chem. 2007, 50, 856.
14. Liang, F.; Kumar, S.; Zhang, Z. Y. Mol. Biosyst. 2007, 3, 308.
15. Wan, Z.-K.; Follows, B.; Kirincich, S.; Wilson, D.; Binnun, E.; Xu, W.; Joseph-
McCarthy, D.; Wu, J.; Smith, M.; Zhang, Y.-L.; Tam, M.; Erbe, D.; Tam, S.; Saiah,
E.; Lee, J. Bioorg. Med. Chem. Lett. 2007, 17, 2913.
16. Shrestha, S.; Bhattarai, B. R.; Chang, K. J.; Lee, K.-H.; Cho, H. Bioorg. Med. Chem.
Lett. 2007, 17, 2760.
17. Dixit, M.; Tripathi, B. K.; Tamrakar, A. K.; Srivastava, A. K.; Kumar, B.; Goel, A.
Bioorg. Med. Chem. 2007, 15, 727.
18. Hansen, S. K.; Cancilla, M. T.; Shiau, T. P.; Kung, J.; Chen, T.; Erlanson, D. A.
Biochemistry 2005, 44, 7704.
19. Praly, J. -P.; He, L.; Qin, B. B.; Tanoh, M.; Chen, G. R. Tetrahedron Lett. 2005, 46,
7081.
20. Puius, Y. A.; Zhao, Y.; Sullivan, M.; Lawrence, D. S.; Almo, S. C.; Zhang, Z. Y. Proc.
Natl. Acad. Sci. U.S.A. 1997, 94, 13420.
21. Wiesmann, C.; Barr, K. J.; Kung, J.; Zhu, J.; Erlanson, D. A.; Shen, W.; Fahr, B. J.;
Zhong, M.; Taylor, L.; Randal, M.; McDowell, R. S.; Hansen, S. K. Nat. Struct. Mol.
Biol. 2004, 11, 730.
22. Hansen, S. K.; Cancilla, M. T.; Shiau, T. P.; Kung, J.; Chen, T.; Erlanson, D. A.
Biochemistry 2005, 44, 7704.
23. Sun, J.-P.; Fedorov, A. A.; Lee, S.-Y.; Guo, X.-L.; Shen, K.; Lawrence, D. S.; Almo, S.
C.; Zhang, Z.-Y. J. Biol. Chem. 2003, 278, 12406.
24. Srinivasan, R.; Uttamchandani, M.; Yao, S. Q. Org. Lett. 2006, 8, 713.
25. Kolb, H. C.; Finn, M. G.; Sharpless, K. B. Angew. Chem., Int. Ed. 2001, 40, 2004.
26. Tornoe, C. W.; Christensen, C.; Meldal, M. J. Org. Chem. 2002, 67, 3057.
27. Isobe, H.; Cho, K.; Solin, N.; Werz, D. B.; Seeberger, P. H.; Nakamura, E. Org. Lett.
2007, 9, 4611.
3.1.10.4. Dimeric b-C-
25. Oxidation of compound 23 (43 mg, 0.035 mmol) afforded
D-galactopyranosyl 1,4-naphthoquinone
25 as a yellow solid (23 mg, 56%). TLC: Rf = 0.58 (petroleum ether/
EtOAc = 1:2), Rf = 0.91 (EtOAc); 1H NMR (400 MHz, CDCl3): d = 8.48
(s, 1H, Ar-H), 8.12–7.96 (m, 6H, Ar-H), 7.73 (m, 5H, 2 ꢂ CH triazole,
Ar-H), 7.30 (t, 1H, J = 8.0 Hz, Ar-H), 7.18 (s, 2H, 2 ꢂ H-3, naphtho-
quinone), 5.59 (d, 2H, J4 ,3 = 2.4 Hz, 2 ꢂ H-40), 5.40 (s, 4H,
2 ꢂ CH2), 5.31–5.21 (m, 4H, 2 ꢂ H-20, 2 ꢂ H-30), 4.86 (d, 2H,
0
0
J1 ,2 = 9.2 Hz, 2 ꢂ H-10), 4.65 (dd, 2H, J6 a,5 = 3.2 Hz, J6 a,6 b = 14.0 Hz,
0
0
0
0
0
0
2 ꢂ H-60a), 4.49 (dd, 2H, J6 b,5 = 8.4 Hz, J6 a, 6 b = 14.0 Hz, 2 ꢂ H-60b),
4.40 (m, 2 H, 2 ꢂ H-50), 2.25, 1.99, 1.87 (3s, 18H, 6 ꢂ COCH3); 13C
NMR (75 MHz, CDCl3): d = 184.6, 183.2, 170.0, 169.8, 169.6, 165.2
(C@O), 142.8 (Cq triazole), 135.8, 134.4, 134.3, 134.0, 131.6,
130.3, 129.8, 128.1, 126.9, 126.3 (Ar), 125.2 (CH triazole), 76.2,
73.9, 72.5, 69.9, 67.8 (C-10 to C-50), 59.0 (CH2), 49.2 (C-60), 20.9,
20.7, 20.4 (COCH3). HRESIMS: calcd for C58H52N6O22: 1207.3032;
found: m/z 1207.3032.
0
0
0
0
3.2. PTP1B inhibitory assay
28. Helms, B.; Mynar, J. L.; Hawker, C. J.; Fréchet, J. M. J. J. Am. Chem. Soc. 2004, 126,
15020.
29. Wu, P.; Malkoch, M.; Hunt, J. N.; Vestberg, R.; Kaltgrad, E.; Finn, M. G.; Fokin, V.
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Biochim. Biophys. Acta 2006, 1760, 1505.
Recombinant human PTP1B catalytic domain was expressed
and purified according to procedures described previously.34 The
enzymatic activities of PTP1B catalytic domain were determined
at 30 °C by monitoring the hydrolysis of pNPP. Dephosphorylation
of pNPP generates product pNP, which can be monitored at
405 nm. In a typical 100 lL assay mixture containing 50 mM
MOPS, pH 6.5, 2 mM pNPP and recombinant enzymes, PTP1B activ-
ities were continuously monitored on a SpectraMax 340 micro-