Synthesis of 2-(β-D-glucopyranosyl)-5-(substituted-amino)-1,3,4-oxa-and -thiadiazoles for the inhibition of glycogen phosphorylase
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G aphical abs ac
pp xxx–xxxSyn hesis o 2-(b-
DD
-glucopy anosyl)-5-(subs i u ed-amino)-1,3,4-oxa- and - hiadiazoles o he inhibi ion
o glycogen phospho ylase
Béla Sz}
ocs, Ma ie a Tó h
*
, Tibo Docsa, Pál Ge gely, László Somsák
*
CAR 6420 No. o Pages 1, Model 5G
26 Ma ch 2013
Highligh s
P epa a ion o new anhyd o-aldose semica bazones and hiosemica bazones. Syn hesis o 2-acylamino- o 2-a ylamino-5-(b-
DD
-glucopy -
anosyl)-1,3,4-oxa- and - hiadiazoles. Low mic omola inhibi o o glycogen phospho ylase.
CN
O
OBz
BzO
BzO
OBz
H
C
O
OH
HO
HO
OH
N
H
N
X
NHR
H
C
O
OBz
BzO
BzO
OBz
N
H
N
X
NHR
O
OH
HO
HO
OH
X
N
N
N
H
O
R'
X=O,S;R'=CH
3
,Ph,2-Naph hyl,4- Bu-C
6
H
4
X=O,S;R=H,Ph,R'CO
X=O,S;R=H,Ph
No inhibi ion a 625 µM.
Bes inhibi io : X = O; R = Ph (K
i
=29µM).
O
OBz
BzO
BzO
OBz
S
N
N
N
H
O
R'
O
R'
*
o
1
Syn hesis o 2-(b-D-glucopy anosyl)-5-(subs i u ed-amino)-1,3,4-oxa-
and - hiadiazoles o he inhibi ion o glycogen phospho ylase
Béla Sz}
ocs
a
,Ma ie a Tó h
a,
⇑
,Tibo Docsa
b
,Pál Ge gely
b
,László Somsák
a,
⇑
a
Depa men o O ganic Chemis y, Uni e si y o Deb ecen, POB 20, H-4010 Deb ecen, Hunga y
b
Depa men o Medical Chemis y, Medical and Heal h Science Cen e, Uni e si y o Deb ecen, Egye em é 1, H-4032 Deb ecen, Hunga y
a icle in o
A icle his o y:
Recei ed 19 Janua y 2013
Recei ed in e ised o m 8 Ma ch 2013
Accep ed 12 Ma ch 2013
A ailable online xxxx
20 Keywo ds:
C-Glycosyl- o maldehyde
( hio)semica bazone
C-Glycosyl-1,3,4-oxadiazole
C-Glycosyl-1,3,4- hiadiazole
Glycogen phospho ylase
Inhibi o
abs ac
O-Pe benzoyla ed 4-phenyl-[C-(b-D-glucopy anosyl) o maldehyde]semica bazone was p epa ed in he
eac ion o O-pe benzoyla ed b-D-glucopy anosyl cyanide and 4-phenylsemica bazide in he p esence
o Raney-Ni. Acyla ion o O-pe benzoyla ed C-(b-D-glucopy anosyl) o maldehyde semica bazone
u nished he co esponding 4-acyl-[C-(b-D-glucopy anosyl) o maldehyde]semica bazones. The eac ion
o O-pe benzoyla ed C-(b-D-glucopy anosyl) o maldehyde semica bazone wi h he co esponding
hiosemica bazide esul ed in O-pe benzoyla ed C-(b-D-glucopy anosyl) o maldehyde hiosemica ba-
zone and i s 4-phenyl de i a i e. Acyla ion o O-pe benzoyla ed C-(b-D-glucopy anosyl) o maldehyde
hiosemica bazone p o ided he co esponding 4-acyl-2-acylamino-5-(b-D-glucopy anosyl)-
D
2
-1,3,4-
hiadiazolidines. Oxida i e ans o ma ions o hese p ecu so s ga e O-p o ec ed 2-(b-D-glucopy ano-
syl)-5-subs i u ed-amino-1,3,4-oxa- and - hiadiazoles. The O-benzoyl p o ec ing g oups we e emo ed
unde base-ca alysed anses e ifica ion condi ions. The C-glucopy anosyl he e ocyclic compounds
p o ed inac i e agains abbi muscle glycogen phospho ylase b, howe e , he semica bazones showed
mode a e inhibi ion (bes inhibi o was 4-phenyl-[C-(b-D-glucopy anosyl) o maldehyde]semica bazone
(K
i
=29
l
M).
Ó2013 Published by Else ie L d.
1. In oduc ion
Glycogen phospho ylase (EC 2.4.1.1) inhibi o s (GPIs) ha e been
conside ed as possible means o he apeu ic in e en ion in ype 2
diabe es and some o he diseased s a es (e.g., ea ly ca diac and ca -
50
dio ascula disease in non-diabe ics, ca diac a hy hmias, ischae-
mic inju ies, and umou g ow h) as a ionalized in he ecen
e iew li e a u e.
1–5
Among se e al compound classes
4,6
glucose
de i a i es a e one o he mos in ensi ely in es iga ed inhibi-
o s.
5,7
The quo ed e iew a icles
5,7
p o ide he eade wi h a
de ailed desc ip ion o he s uc u e–ac i i y ela ionships (SAR)
o glucose analogue GPIs, he e o e, only hose ea u es a e sum-
ma ized he eina e which o m he di ec basis o ou compound
design. N-Acyl-b-
D
-glucopy anosylamines
8
(Cha 1,I:e.g. o
R = 2-naph hyl K
i
measu ed agains abbi muscle GPb(RMGPb)
9
60
was 10–13
l
M,
8,10
)N-a yl-N
0
-b-
D
-glucopy anosyl u eas
4,11
II
(R = 2-naph hyl: K
i
(RMGPb) 5.2
l
M), as well as N-acyl-N
0
-b-
D
-glu-
copy anosyl u ea de i a i es
4,12
III (R = 2-naph hyl: K
i
(RMGPb)
0.35
l
M) ha e been shown o inhibi he enzyme in he low mic o-
mola ange. As a pa o a p og amme o eplace he NHCO moie y
o he abo e compounds by non-classical bioisos e ic he e ocyclic
linke s, among o he s we ha e ca ied ou he syn hesis o
compounds IA.
13
Enzyma ic es s as well as c ys allog aphic s ud-
ies e ealed high simila i y o he amide (see K
i
o Iabo e) and he
1,2,3- iazole ype ( o IA R = 2-naph hyl: K
i
(RMGPb) 16
l
M)
70
inhibi o s bo h in binding s eng h and s uc u al ea u es o he
enzyme–inhibi o complexes.
10
Applying he isome ic oxadiazoles
B,D, and Eas linke s esul ed in inhibi o s o a ying e ficiency,
whe eby he 3-a yl-5-b-
D
-glucopy anosyl-1,2,4-oxadiazole (IE
ype) de i a i es p o ed o be he mos po en compounds ( o
he bes inhibi o whe e R = 2-naph hyl he K
i
(RMGPb) was
2.4
l
M).
14,15
He ein we epo on he syn hesis and enzyma ic es o
compounds o ype II and III wi h 1,3,4-oxadiazole Band 1,3,4-
hiadiazole Cas linke s ep esen ing bioisos e ic eplacemen s o
80
NHCO moie ies o N-subs i u ed-N
0
-b-
D
-glucopy anosyl u eas.
These s udies con ibu e o an ex ension o SAR ela ed o in e ac-
ions o inhibi o s in he b-channel o he enzyme which can
accommoda e aglycons o he glucose analogue compounds and
is lined wi h amino acid side chains o mixed cha ac e .
16
2. Resul s and discussion
Fo he o ma ion o he he e ocyclic pa s o he a ge com-
pounds oxida i e ing closu e o ( hio)semica bazones (simila o
0008-6215/$ - see on ma e Ó2013 Published by Else ie L d.
h p://dx.doi.o g/10.1016/j.ca es.2013.03.009
⇑
Co esponding au ho s. Tel.: +36 52512900x22474; ax: +36 52512744 (M.T.);
el.: +36 52512900x22348; ax: +36 52512744 (L.S.).
E-mail add esses: [email p o ec ed] (M. Tó h), somsak.laszlo@
science.unideb.hu (L. Somsák).
Q1
Q2
Ca bohyd a e Resea ch xxx (2013) xxx–xxx
Con en s lis s a ailable a SciVe se ScienceDi ec
Ca bohyd a e Resea ch
jou nal homepage: www.else ie .com/loca e/ca es
CAR 6420 No. o Pages 10, Model 5G
27 Ma ch 2013
Please ci e his a icle in p ess as: Sz}
ocs, B.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.03.009
ha applied o he syn heses o IB
15
) was en isaged as he key
s ep. To his end, he p o ec ed C-(b-
D
-glucopy anosyl) o malde-
90
hyde semica bazone 2was p epa ed by a eac ion o NH
2
C
(@O)NHNH
2
wi h O-pe benzoyla ed b-
D
-glucopy anosyl cyanide
17
1in he p esence o Raney Ni and NaH
2
PO
2
a 40 °C(Scheme 1)
as desc ibed ea lie .
18
O-Pe benzoyla ed 4-phenyl-[C-(b-
D
-gluco-
py anosyl) o maldehyde]semica bazone 3was ob ained in a
simila way using PhNHC(@O)NHNH
2
as he apping agen .
Debenzoyla ion was pe o med by he Zemplén p o ocol o esul
in mode a e yields o semica bazones 6and 7.
In o de o ge hiosemica bazones 4and 5, acid ca alysed
ansimina ion o 2was ca ied ou because he di ec ans o ma-
100
iono b-
D
-glucopy anosylcyanide1in o hiosemica bazones ailed.
Thus, semica bazone 2was eac ed wi h NH
2
C(@S)NHNH
2
o
PhNHC(@S)NHNH
2
in glacial AcOH a 70 °C o gi e he co espond-
ing hiosemica bazones 4and 5, espec i ely. Remo al o he ben-
zoyl p o ec ing g oups in 4and 5was ca ied ou by LiOH in MeOH
o gi e compounds 8and 9, espec i ely, in sa is ac o y yields.
To ob ain p ecu so s o compounds IIIB semica bazone 2was
ea ed by acid chlo ides in CH
2
Cl
2
in he p esence o E
3
N o gi e
O-pe benzoyla ed 4-acyl-[C-(b-
D
-glucopy anosyl) o maldehyde]
semica bazones 12–15 in sa is ac o y yields (Scheme 2). Unde
110
hese condi ions no eac ion ook place wi h hiosemica bazone
3. Acyla ion o 3in d y py idine a 80 °C esul ed in he o ma ion
o 4-acyl-2-acylamino-5-(b-
D
-glucopy anosyl)-
D
2
-1,3,4- hiadiazo-
lines 16–19 as insepa able mix u es o wo dias e eoisome s in
good yields. This obse a ion is in acco d wi h he li e a u e
expe iences.
19
Ring closing eac ions o he p ecu so s unde oxida i e
condi ions we e s udied nex . Semica bazone 3was eac ed wi h
Pb (OAc)
4
in glacial AcOH a 70 °C o u nish O-pe benzoyla ed
2-phenylamino-5-(2,3,4,6- e a-O-benzoyl-b-
D
-glucopy anosyl)-1,
CN
O
OBz
BzO
BzO
OBz
H
C
O
OBz
BzO
BzO
OBz
N
H
N
X
NHR H
C
O
OH
HO
HO
OH
N
H
N
X
NHR
1
2R=H, X=O(90%)
3R=Ph,X=O(64%)
4R=H, X=S(85%)
5R=Ph,X=S(87%)
6R=H, X=O(63%)
7R=Ph,X=O(48%)
8R=H, X=S(95%)
9R=Ph,X=S(77%)
a
b
c
d
d
c
b
Scheme 1. Reagen s and condi ions: (a) RNHC(@O)NHNH
2
, Raney Ni, NaH
2
PO
2
,H
2
O, AcOH, py idine, 40 °C; (b) RNHC(@S)NHNH
2
, AcOH, 70 °C; (c) NaOMe, d y MeOH, 25 °C;
(d) LiOH, d y MeOH, 0 °C.
H
C
O
OBz
BzO
BzO
OBz
N
H
N
X
NHR
2R=H, X=O
3R=Ph,X=O
4R=H, X=S
H
C
O
OBz
BzO
BzO
OBz
N
H
N
O
H
N
O
RO
OBz
BzO
BzO
OBz
X
NN
N
H
O
R
O
OBz
BzO
BzO
OBz
O
N
N
N
H
Ph
11 (97 %)
O
OH
HO
HO
OH
X
NN
N
H
O
R
a
om 3
X=O X=S X=O X=S
R=CH
3
12 (47 %) 20 (50 %) 24 (67 %) 28 (41 %) 32 (92 %)
R=Ph 13 (70 %) 21 (57 %) 25 (68 %) 29 (87 %) 33 (68 %)
R = 2-Naph hyl 14 (50 %) 22 (69 %) 26 (59 %) 30 (93 %) 34 (57 %)
R=4- Bu-C
6
H
4
15 (55 %) 23 (60 %) 27 (84 %) 31 (76 %) 35 (72 %)
c
o 12-15 om 4
O
OH
HO
HO
OH
O
NN
N
H
Ph
b
10 (45 %)
be
O
OBz
BzO
BzO
OBz
S
NN
N
H
O
R
O
R16 R=CH
3
(77%, 95:5
a
)
17 R = Ph (62%, 72:18
a
)
18 R=2-Naph hyl(62%,85:15
a
)
19 R=4- Bu-C
6
H
4
(38%, 1 isome )
d
e
12-15 20-27 28-35
a
Dias e eome ic a io.
*
Scheme 2. Reagen s and condi ions: (a) Pb(OAc)
4
, AcOH, 70 °C; (b) NaOMe, d y MeOH, 25 °C; (c) RCOCl, E
3
N, d y CH
2
Cl
2
; (d) RCOCl, d y py idine, 80 °C; (e) PIDA, CH
2
Cl
2,
25 °C.
O
HO
HO
HO
OH H
NR
O
linke
I
O
HO
HO
HO
OH H
NH
N
O
linke
II
R
linke s o eplace he
highligh ed NHCO moie ies
N
N
N
N
X
N
N
N
O
O
N
N
A
BX=O
CX=S
D
O
HO
HO
HO
OH H
NH
N
O O
R
III
linke
E
Cha 1.
2B. Sz}
ocs e al. / Ca bohyd a e Resea ch xxx (2013) xxx–xxx
CAR 6420 No. o Pages 10, Model 5G
27 Ma ch 2013
Please ci e his a icle in p ess as: Sz}
ocs, B.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.03.009
120
3,4-ox adiazole 10 (Scheme 2). T ials o use o he oxidizing agen s
such as FeCl
3
o PIDA ailed, 3was eco e ed in hese eac ions.
Dep o ec ion o 10 was achie ed by he Zemplén me hod o gi e
11 in an excellen yield. Thiosemica bazone 4 esis ed se e al
oxida ion eagen s (PIDA, Pb(OAc)
4
, FeCl
3
,B
2
,and K
3
Fe(CN)
6
).
Semica bazones 12–15 we e eac ed wi h PIDA in CH
2
Cl
2
a o
ge O-pe benzoyla ed 2-acylamino-5-(b-
D
-glucopy anosyl)-1,3,4-
oxadiazoles 20–23 in good yields (Scheme 2). Thiadiazolines 16–
19 we e oxidized in a simila way o esul in 1,3,4- hiadiazoles
24–27.Debenzoyla ions we e pe o med by he Zemplén p o ocol
130
o gi e good and excellen yields o 1,3,4-oxadiazoles 28–31 and
1,3,4- hiadiazoles 32–35.
The dep o ec ed compounds we e es ed o hei inhibi ion
po ency agains abbi muscle glycogen phospho ylase b(RMGPb)
acco ding o he p o ocol desc ibed ea lie .
20
The esul s a e sum-
ma ized in Table 1 also showing he inhibi o y e ficiency o some
ele an e e ence compounds.
Replacemen o he NHCO moie y in compound ype II (en y 1)
by a 1,3,4-oxadiazole ing (11) esul ed in a loss o inhibi ion. A
simila obse a ion was made wi h compounds ype III whe eby
140
on changing he NHCO g oup o he acylu ea de i a i es (en y
2) o ei he 1,3,4-oxadiazole (en y 3, 28–31) o 1,3,4- hiadiazole
(en y 4, 32–35) he e ficiency was los . These findings esemble
hose ob ained wi h compounds ype IB in Cha 1 whe e 1,3,4-
oxadiazole eplacemen s in N-acyl-glucopy anosylamines esul ed
in p ac ically inac i e compounds.
15
I ollows om hese esul s
ha he 1,3,4-oxadiazole and 1,3,4- hiadiazole moie ies canno
con ibu e o a ou able in e ac ions in he b-channel o RMGPb.
In e es ingly, semica bazones 6and 7(en y 6) and hiosemi-
ca bazones 8and 9(en y 7), he ‘open chain’ p ecu so s o he
150
a ge compounds o his wo k, showed week o mode a e inhibi-
ion. Semica bazones 6and 7a e be e inhibi o s han he hio-
semica bazone coun e pa s 8and 9. Phenyl de i a i es 7and 9
a e mo e e ficien han he unsubs i u ed compounds 6and 8,
espec i ely, and his is in ag eemen wi h he gene al end o
show s onge inhibi ion by hose compounds which ha e a la -
ge( ) a oma ic pa in he aglycon.
4
A compa ison o 7(ac ually
he bes inhibi o in his s udy) o he biu e ype inhibi o (en y
5) indica es no significan di e ence in he inhibi ion cons an s.
This may e eal ha in e ac ions o he whole NHCO linke moie y
160
o i s ca bonyl g oup has p obably less significance in binding o
he enzyme han hose o he ‘second’ NHCO uni .
3. Conclusion
The educ i e ans o ma ion o
D
-glycopy anosyl cyanides in
he p esence o acylhyd azines was ex ended o he p epa a ion
o new anhyd o-aldose semica bazone and hiosemica bazone
de i a i es. Acyla ion o semica bazones yielded he expec ed
4-acyl semica bazones, while unde simila condi ions hiosemi-
ca bazones ga e O-pe acyla ed 4-acyl-2-acylamino-5-(b-
D
-gluco-
py anosyl)-
D
2
-1,3,4- hiadiazolines. Oxida ion o he a o emen
170
ioned p ecu so s esul ed in p o ec ed 2-(b-
D
-glucopy anosyl)-5-
(subs i u ed-amino)-1,3,4-oxa- and hiadiazoles, espec i ely.
A e O-dep o ec ion enzyme kine ic measu emen s showed he
1,3,4-oxa- and - hiadiazoles o be inac i e agains abbi muscle
Table 1
Inhibi ion o abbi muscle glycogen phospho ylase b (RMGPb) by selec ed glucose de i a i es and he new compounds (K
i
[
l
M])
O
HO
HO
HO
OH
linke H
NR
II
En y Linke R Linke R
1 NHCO
O
N
N
18
4
11 No inh.
a
O
HO
HO
HO
OH
linke
III
H
N
O
R
R
En y Linke CH
3
2 NHCO 305
21
4.6
21
0.35
4
0.7
4
3
O
N
N
28 No inh.
a
29 No inh.
a
30 No inh.
a
31 No inh.
a
4
N
S
N
32 No inh.
a
33 No inh.
a
34 No inh.
a
35 No inh.
a
O
HO
HO
HO
OH
linke
IV
H
N
X
NHR
R
En y Linke X H
5 NHCO O — 21
4
6CH@NO6332
b
729
7CH@NS 8No inh. 9300
b
a
No inhibi ion a a es ed concen a ion o 625
l
M.
b
Calcula ed om he IC
50
alue by using a web-based ool.
22
B. Sz}
ocs e al. / Ca bohyd a e Resea ch xxx (2013) xxx–xxx 3
CAR 6420 No. o Pages 10, Model 5G
27 Ma ch 2013
Please ci e his a icle in p ess as: Sz}
ocs, B.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.03.009
glycogen phospho ylase b. Howe e , he p ecu so ‘open chain’ 4-
phenyl semica bazone p o ed o be a low mic omola GPI, equipo-
en wi h a biu e de i a i e o simila chain leng h be ween he
suga and he a oma ic pa o he molecule. Replacemen o he
oxygen by a sul u a om caused he loss o ac i i y. This la e find-
ing may indica e a di e ence in con ibu ion o he binding o he
180
wo NHCO uni es o N-acyl-N
0
-glucopy anosyl u ea ype inhibi o s
o glycogen phospho ylase.
4. Expe imen al
4.1. Gene al me hods
Mel ing poin s we e measu ed in open capilla y ubes o on a
Kofle ho -s age and a e unco ec ed. Op ical o a ions we e
de e mined wi h a Pe kin–Elme 241 pola ime e a oom empe -
a u e. NMR spec a we e eco ded wi h B uke 360 (360/90 MHz
o
1
H/
13
C) o B uke 400 (400/100 MHz o
1
H/
13
C) spec ome e s.
Chemical shi s a e e e enced o TMS as he in e nal e e ence
190
(
1
H), o o he esidual sol en signals (
13
C). Mic oanalyses
we e pe o med on an Elemen a a io Mic o cube. ESI-MS we e
eco ded wi h a The mo Scien ific LTQ XL ins umen . TLC was
pe o med on DC-Alu olle Kieselgel 60 F
254
(Me ck). TLC pla es
we e isualized unde UV ligh , and by gen le hea ing wi h a
comme cially a ailable hea gun wi hou any cha ing eagen .
Fo column ch oma og aphy Kieselgel 60 (Me ck, pa icle size
(0.063–0.200 mm) was applied. O ganic solu ions we e d ied o e
anhyd ous MgSO
4
, and concen a ed unde diminished p essu e a
40–50 °C (wa e ba h).
200
4.2. 4-Phenyl-[C-(2,3,4,6- e a-O-benzoyl-b-
D
-glucopy anosyl)
o maldehyde]semica bazone (3)
Raney Ni (3.53 g, om an aqueous suspension, Me ck) was
added a o a igo ously s i ed solu ion o py idine (14 mL),
H
2
O (9 mL), and AcOH (9 mL). Then, NaH
2
PO
2
(1.76 g, 20.0 mmol),
(2,3,4,6- e a-O-benzoyl-b-
D
-glucopy anosyl) cyanide
17
(1, 1.50 g,
2.48 mmol), and PhNHC(@O)NHNH
2
(0.75 g, 4.96 mmol) we e
added o he mix u e. The eac ion mix u e was s i ed and hea ed
a 40 °C. When he eac ion was comple e (TLC, 1:2 E OAc/hexane)
he insoluble ma e ials we e fil e ed o wi h suc ion, and washed
210
wi h E OAc (3 15 mL). The o ganic laye o he fil a e was sepa-
a ed, washed wi h 10% HCl (2 15 mL), sa u a ed NaHCO
3
solu-
ion (3 20 mL), H
2
O(215 mL), d ied, and concen a ed unde
educed p essu e. T aces o py idine we e emo ed by epea ed
co-e apo a ions wi h oluene. The c ude p oduc was pu ified by
column ch oma og aphy (2:3 E OAc/hexane) o yield 1.17 g (64%)
o 3as a whi e amo phous p oduc . [
a
]
D
= +33 (c0.52, CHCl
3
); R
:
0.44 (1:1 E OAc/hexane);
1
H NMR (CDCl
3
, 360 MHz) d(ppm)
10.36 (1H, s, NH), 8.20 (1H, s, NH), 8.05–7.02 (26H, m, A , CH@N),
6.08, 6.03, 5.77 (3H, 3 p , J= 9.2, 9.5 Hz in each, H-2, H-3, H-4), 4.67
220
(1H, dd, J
6a,6b
= 12.7 Hz, H-6a), 4.53 (1H, dd, J
1,CH
@
N
= 4.1 Hz,
J
1,2
= 9.1 Hz, H-1), 4.40 (1H, dd, H-6b), 4.25 (1H, ddd,
J
5,6a
= 3.0 Hz, J
5,6b
= 4.5 Hz, J
4,5
= 9.9 Hz, H-5).
13
C NMR (CDCl
3
,
360 MHz) d(ppm) 166.0, 165.8, 165.0 (CO), 153.7 (NHCONH),
138.0 (CH@N), 136.5–119.4 (A ), 76.3, 76.2, 74.3, 69.3 (C-1 o
C-5), 63.0 (C-6). Anal. Calcd o C
42
H
35
N
3
O
10
(741.74): C, 68.01,
H, 4.76; N, 5.67. Found: C, 69.00; H, 4.83; N, 5.62.
4.3. C-(2,3,4,6-Te a-O-benzoyl-b-
D
-glucopy anosyl)
o maldehyde hiosemica bazone (4)
C-(2,3,4,6-Te a-O-benzoyl-b-
D
-glucopy anosyl) o maldehyde
230
semica bazone
18
(2, 1.50 g, 2.25 mmol) was dissol ed in glacial
AcOH (65 mL) and hen NH
2
C(@S)NHNH
2
(0.83 g, 9.05 mmol)
was added. The mix u e was s i ed and hea ed a 70 °C. The eac-
ion was moni o ed by TLC (1:1 E OAc/hexane). When he eac ion
was comple e, he eac ion mix u e was dilu ed wi h H
2
O
(200 mL), and ex ac ed wi h E OAc (4 20 mL), and washed wi h
NaHCO
3
(3 25 mL), and H
2
O(120 mL). The o ganic phase was
d ied (MgSO
4
), and e apo a ed unde educed p essu e. The c ude
p oduc was pu ified by column ch oma og aphy (1:1 E OAc/hex-
ane) o gi e 1.30 g (85%) o 4as a yellow amo phous p oduc 4.
240
[
a
]
D
= +11 (c0.35, CHCl
3
); R
: 0.33 (1:2 E OAc/hexane);
1
HNMR
(CDCl
3
, 360 MHz) d(ppm) 9.94 (1H, s, NH), 8.07–7.10 (21H, m,
A , CH@N), 6.34 (1H, s, J= 2.4 Hz, NH), 5.98, 5.87, 5.71 (3H, 3 p ,
J= 9.5, 9.8 Hz in each, H-2, H-3, H-4), 4.66 (1H, dd, J
6a,6b
= 12.5 Hz,
H-6a), 4.49–4.43 (2H, m, H-1, H-6b), 4.22 (1H, ddd, J
5,6a
= 2.9 Hz,
J
5,6b
= 5.0 Hz, J
4,5
= 9.9 Hz, H-5).
13
C NMR (CDCl
3
, 360 MHz) d
(ppm) 179.2 (CS), 166.1, 165.9, 165.8, 165.1 (CO), 139.2 (CH@N),
133.7–128.3 (A ), 76.9, 76.4, 73.9, 69.6, 69.1 (C-1 o C-5), 62.9
(C-6). Anal. Calcd o C
36
H
31
N
3
O
9
S (681.71): C, 63.43, H, 4.58; N,
6.16; S, 4.70. Found: C, 63.54; H, 4.67; N, 6.09; S, 4.78.
250
4.4. 4-Phenyl-[C-(2,3,4,6- e a-O-benzoyl-b-
D
-
glucopy anosyl) o maldehyde] hiosemica bazone (5)
C-(2,3,4,6-Te a-O-benzoyl-b-
D
-glucopy anosyl) o maldehyde
semica bazone
18
(2, 1.00 g, 1.50 mmol) was dissol ed in glacial
AcOH (43 mL) and hen PhNHC(@S)NHNH
2
(0.83 g, 9.05 mmol)
was added. The mix u e was s i ed and hea ed a 70 °C. The eac-
ion was moni o ed by TLC (1:1 E OAc/hexane). When he eac ion
was comple e, he eac ion mix u e was dilu ed wi h H
2
O
(150 mL), and ex ac ed wi h E OAc (4 15 mL), and washed wi h
NaHCO
3
(3 20 mL), and H
2
O(115 mL). The o ganic phase was
260
d ied (MgSO
4
), and e apo a ed unde educed p essu e. The c ude
p oduc was pu ified by column ch oma og aphy (1:1.5 E OAc/
hexane) o yield 1.00 g (87%) o 5as a yellow amo phous p oduc .
[
a
]
D
=+7 (c0.33, CHCl
3
); R
: 0.55 (1:1 E OAc/hexane);
1
HNMR
(CDCl
3
, 360 MHz) d(ppm) 10.74 (1H, s, NH), 9.19 (1H, s, NH),
8.04–7.18 (26H, m, A , CH@N), 6.09–5.99 (2H, m, H-2 and/o H-3
and/o H-4), 5.76 (1H, 1 p , J= 9.3 Hz, H-2 o H-3 o H-4), 4.66
(1H, dd, J
5,6a
= 2.4 Hz, J
6a,6b
= 12.2 Hz, H-6a), 4.54 (1H, dd, J
1,
CH
@
N
= 4.0 Hz, J
1,2
= 9.2 Hz, H-1), 4.49 (1H, dd, J
5,6b
= 5.3 Hz,
H-6b), 4.26–4.22 (1H, m, H-5).
13
C NMR (CDCl
3
, 360 MHz) d
270
(ppm) 176.0 (CS), 166.0, 165.2, 165.2, 164.9 (CO), 138.1 (CH@N),
137.6–124.5 (A ), 76.0, 76.1, 74.1, 69.33, 69.1 (C-1 o C-5), 62.8
(C-6). Anal. Calcd o C
42
H
35
N
3
O
9
S (757.81): C, 66.57, H, 4.66; N,
5.54; S, 4.23. Found: C, 66.63; H, 4.72; N, 5.58; S, 4.29.
4.5. 2-Phenylamino-5-(2,3,4,6- e a-O-benzoyl-b-
D
-
glucopy anosyl)-1,3,4-oxadiazole (10)
Semica bazone 3(0.20 g, 0.27 mmol) was dissol ed in glacial
AcOH (3 mL). Then Pb(OAc)
4
(0.18 g, 0.40 mmol) was added, and
he mix u e was s i ed and hea ed a 70 °C. The eac ion was
moni o ed by TLC (1:2 E OAc/hexane). When he eac ion was com-
280
ple e, he eac ion mix u e was dilu ed wi h H
2
O (15 mL), and ex-
ac ed wi h E OAc (3 6 ml). The o ganic phase was d ied
(MgSO
4
), and e apo a ed unde educed p essu e. The c ude p od-
uc was pu ified by column ch oma og aphy (1:2 E OAc/hexane)
o yield 90 mg (45%) o 10 as a whi e amo phous p oduc . [
a
]
D
=–
17 (c0.21, CHCl
3
); R
: 0.41 (1:2 E OAc/hexane);
1
H NMR (CDCl
3
,
360 MHz) d(ppm) 8.87 (1H, s, NH), 8.02–7.00 (25H, m, A ), 6.08,
5.93, 5.83 (3H, 3 p , J= 9.5, 9.7 Hz in each, H-2, H-3, H-4), 5.13
(1H, d, J
1,2
= 9.9 Hz, H-1), 4.65 (1H, dd, J
5,6a
= 2.0 Hz, J
6a,6b
= 12.4 Hz,
H-6a), 4.51 (1H, dd, J
5,6b
= 5.2 Hz, H-6b) 4.40–4.30 (1H, m, H-5).
13
C
290
NMR (CDCl
3
, 360 MHz) d(ppm) 166.1, 165.7, 165.1, 164.9 (CO),
161.2, 154.8 (C-oxadiazole), 137.4–117.8 (A ), 76.8, 73.6, 71.9,
70.1, 69.1 (C-1 o C-5), 63.0 (C-6). Anal. Calcd o C
42
H
33
N
3
O
10
(739.73): C, 68.19, H, 4.50; N, 5.68. Found: C, 68.25; H, 4.59; N, 5.61.
4B. Sz}
ocs e al. / Ca bohyd a e Resea ch xxx (2013) xxx–xxx
CAR 6420 No. o Pages 10, Model 5G
27 Ma ch 2013
Please ci e his a icle in p ess as: Sz}
ocs, B.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.03.009
4.6. Gene al p ocedu e I o he syn hesis o O-pe benzoyla ed
4-acyl-[C-(b-
D
-glucopy anosyl) o maldehyde]semica bazones
C-(2,3,4,6-Te a-O-benzoyl-b-
D
-glucopy anosyl) o maldehyde
semica bazone
18
(2, 0.10 g, 0.15 mmol) was dissol ed in d y
CH
2
Cl
2
(5 mL), and hen E
3
N (0.063 mL, 0.45 mmol) and RCOCl
(0.45 mmol) we e added. The mix u e was s i ed and hea ed a
300
40 °C. The eac ion was moni o ed by TLC (1:1 E OAc/hexane).
When he eac ion was comple e, he sol en was e apo a ed,
and he esidue was pu ified by column ch oma og aphy.
4.6.1. 4-Ace yl-[C-(2,3,4,6- e a-O-benzoyl-b-
D
-
glucopy anosyl) o maldehyde]semica bazone (12)
F om 2(0.19 g, 0.29 mmol) and AcCl (62
l
L, 0.87 mmol) acco d-
ing o Gene al p ocedu e I (Sec ion 4.6). Pu ified by column ch o-
ma og aphy (1:1.5 E OAc/hexane) o yield 91 mg (47%) o 12 as a
yellow amo phous p oduc . [
a
]
D
=+8(c0.35, CHCl
3
); R
: 0.37 (1:1
E OAc/hexane);
1
H NMR (CDCl
3
, 360 MHz) d(ppm) 9.83 (1H, s,
310
NH), 8.10–7.80 (8H, m, A ), 7.58–7.18 (14H, m, A , CH@N, NH),
6.00, 5.73, 5.66 (3H, 3 p , J= 9.6, 9.7 Hz in each, H-2, H-3, H-4),
4.65 (1H, dd, J
5,6a
= 1.8 Hz, J
6a,6b
= 12.2 Hz, H-6a), 4.52–4.37 (2H,
m, H-1, H-6b), 4.28–4.17 (1H, m, H-5), 1.91 (3H, s, CH
3
).
13
C NMR
(CDCl
3
, 360 MHz) d(ppm) 173.8, 166.1, 165.8, 165.3, 165.1, 158.4
(CO, NHCO, NHCONH), 140.5 (CH@N), 133.4–128.2 (A ), 77.7,
76.2, 73.7, 70.2, 69.3 (C-1 o C-5), 63.0 (C-6), 19.7 (CH
3
). Anal. Calcd
o C
38
H
33
N
3
O
11
(707.68): C, 64.49, H, 4.70; N, 5.94. Found: C,
64.59; H, 4.79; N, 5.86.
4.6.2. 4-Benzoyl-[C-(2,3,4,6- e a-O-benzoyl-b-
D
-
320
glucopy anosyl) o maldehyde]semica bazone (13)
F om 2(0.10 g, 0.15 mmol) and BzCl (53
l
L, 0.45 mmol) acco d-
ing o Gene al p ocedu e I (Sec ion 4.6). Pu ified by column ch o-
ma og aphy (1:1.5 E OAc/hexane) o yield 81 mg (70%) o 13 as a
yellow amo phous p oduc . [
a
]
D
= –61 (c0.18, CHCl
3
); R
: 0.51
(1:1 E OAc/hexane);
1
H NMR (CD
3
CN, 360 MHz) d(ppm) 10.28
(1H, b s, NH), 8.06–7.30 (27H, m, A , CH@N, NH), 6.06, 5.80, 5.73
(3H, 3 p , J= 9.5, 9.8 Hz in each, H-2, H-3, H-4), 4.70–4.65 (1H, m,
H-5), 4.59 (2H, dd, J
5,6a
= 2.4 Hz, J
6a,6b
= 12.5 Hz, H-6a), 4.53 (1H,
dd, J
5,6b
= 4.1 Hz, H-6b), 4.42 (1H, d, J
1,2
= 9.1 Hz, H-1).
13
C NMR
330
(CD
3
CN, 360 MHz) d(ppm) 166.7, 166.4, 166.1, 166.0, 164.5 (CO,
NHCO, NHCONH), 145.9 (CH@N) 134.5–118.2 (A ), 78.6, 76.5,
75.1, 71.5, 70.0 (C-1 o C-5), 63.7 (C-6). Anal. Calcd o
C
43
H
35
N
3
O
11
(769.75): C, 67.09, H, 4.58; N, 5.46. Found: C, 67.01;
H, 4.50; N, 5.55.
4.6.3. 4-(2-Naph hoyl)-[C-(2,3,4,6- e a-O-benzoyl-b-
D
-
glucopy anosyl) o maldehyde]semica bazone (14)
F om 2(0.20 g, 0.30 mmol) and 2-naph hoyl chlo ide (172 mg,
0.90 mmol) acco ding o Gene al p ocedu e I (Sec ion 4.6). Pu ified
by column ch oma og aphy (1:3,5 E OAc/ oluene) o yield 123 mg
340
(50%) o 14 as a yellow amo phous p oduc . [
a
]
D
= –134 (c0.16,
CHCl
3
); R
: 0.43 (1:3.5 E OAc/ oluene);
1
H NMR (CDCl
3
, 360 MHz)
d(ppm) 9.79 (1H, b s, NH), 8.18–7.23 (29H, m, A , CH@N, NH),
6.06, 5.77, 5.65 (3H, 3 p , J= 8.7 Hz, 9.3 Hz in each, H-2, H-3, H-4),
4.73–4.58 (2H, m, H-1, H-6a), 4.48 (1H, dd, J
5,6b
= 4.9 Hz,
J
6a,6b
= 12.1 Hz, H-6b), 4.32–4.19 (1H, m, H-5).
13
C NMR (CDCl
3
,
360 MHz) d(ppm) 166.1, 165.7, 165.3, 164.3 (CO, NHCO, NHCONH),
145.1 (CH@N), 134.8–123.8 (A ), 78.5, 76.3, 73.6, 70.8, 69.4 (C-1 o
C-5), 63.2 (C-6). Anal. Calcd o C
47
H
37
N
3
O
11
(819.81): C, 68.86, H,
4.55; N, 5.13. Found: C, 68.97; H, 4.65; N, 5.20.
350
4.6.4. 4-(4- e -Bu ylbenzoyl)-[C-(2,3,4,6- e a-O-benzoyl-b-
D
-
glucopy anosyl) o maldehyde]semica bazone (15)
F om 2(0.10 g, 0.15 mmol) and 4- e -bu ylbenzoyl chlo ide
(88
l
L, 0.45 mmol) acco ding o Gene al p ocedu e I (Sec ion
4.6). Pu ified by column ch oma og aphy (1:2 E OAc/hexane) o
yield 68 mg (55%) o 15 as a yellow amo phous p oduc .
[
a
]
D
= +41 (c0.52, CHCl
3
); R
: 0.27 (1:2 E OAc/hexane);
1
HNMR
(CDCl
3
, 360 MHz) d(ppm) 9.66 (1H, b s, NH), 8.01–7.21 (26H, m,
A , CH@N, NH), 6.06, 5.77, 5.64 (3H, 3 p , J= 8.6, 9.3 Hz in each,
H-2, H-3, H-4), 4.76–4.56 (2H, m, H-1, H-6a), 4.49 (1H, dd,
360
J
5,6b
= 4.9 Hz, J
6a,6b
= 12.1 Hz, H-6b), 4.32–4.18 (1H, m, H-5), 1.26
(9H, s, CH
3
).
13
C NMR (CDCl
3
, 360 MHz) d(ppm) 166.0, 165.9,
165.6, 165.2 (CO, NHCO, NHCONH), 144.6 (CH@N), 155.4, 133.4–
125.2 (A ), 78.6, 76.2, 73.6, 70.8, 69.3 (C-1 o C-5), 63.2 (C-6),
34.7 (C Bu), 30.9 (CH
3
). Anal. Calcd o C
47
H
43
N
3
O
11
(825.86): C,
68.35, H, 5.25; N, 5.09. Found: C, 68.44; H, 5.36; N, 5.16.
4.7. Gene al p ocedu e II o he syn hesis o 4-acyl-2-
acylamino-5-(2,3,4,6- e a-O-benzoyl-b-
D
-glucopy anosyl)-
D
2
-
1,3,4- hiadiazolines
C-(2,3,4,6-Te a-O-benzoyl-b-
D
-glucopy anosyl) o maldehyde
370
hiosemica bazone (4, 0.10 g, 0.14 mmol) was dissol ed in d y py -
idine (5 mL) and hen RCOCl (0.44 mmol) was added. The mix u e
was s i ed and hea ed a 80 °C. The eac ion was moni o ed by TLC
(1:2 E OAc/hexane). When he eac ion was comple e, he sol en
was e apo a ed, and he esidue was pu ified by column
ch oma og aphy.
4.7.1. 2-Ace amido-4-ace yl-5-(2,3,4,6- e a-O-benzoyl-b-
D
-
glucopy anosyl)-
D
2
-1,3,4- hiadiazoline (16)
F om 4(0.20 g, 0.28 mmol) and AcCl (60
l
L, 0.84 mmol) acco d-
ing o Gene al p ocedu e II (Sec ion 4.7). Pu ified by column ch o-
380
ma og aphy (1:2 E OAc/ oluene) o yield 170 mg (77%) o 16 as a
whi e amo phous p oduc . [
a
]
D
= –178 (c0.35, CHCl
3
); R
: 0.30
(1:2 E OAc/ oluene); ESI-MS (posi i e mode) m/z:766.25 [M+H]
+
.
Anal. Calcd o C
40
H
35
N
3
O
11
S (765.78): C, 62.74, H, 4.61; N, 5.49;
S, 4.19. Found: C, 62.71; H, 4.65; N, 5.46; S, 4.15.
majo isome :
1
H NMR (CDCl
3
, 360 MHz) d(ppm) 9.43 (1H, s,
NH), 8.10–7.11 (20H, m, A ), 6.05–5.96 (2H, m, H-2 o H-3 o
H-4, CH- hiadiazoline), 5.68, 5.56 (2H, 2 p , J= 9.8 Hz in each,
H-2 and/o H-3 and/o H-4), 4.53–4.42 (3H, m, H-6a, H-6b, H-1),
4.15–4.08 (1H, m, H-5), 1.99, 1.93 (6H, 2 s, CH
3
).
13
C NMR (CDCl
3
,
390
360 MHz) d(ppm) 169.0, 166.0, 165.6, 165.0 (CO, NHCO), 147.4
(Cq- hiadiazoline), 133.6–125.1 (A ), 76.0, 75.2, 73.5, 69.1, 66.3
(C-1 o C-5), 69.1 (CH- hiadiazoline), 62.4 (C-6), 22.6, 21.5 (CH
3
).
Mino isome :
1
H NMR (CDCl
3
, 360 MHz) d(ppm) 9.14 (1H, s,
NH), 5.84, 5.43 (2H, 2 p , J= 9.5, 9.7 Hz in each, H-2 and/o H-3
and/o H-4).
4.7.2. 2-Benzamido-4-benzoyl-5-(2,3,4,6- e a-O-benzoyl-b-
D
-
glucopy anosyl)-
D
2
-1,3,4- hiadiazoline (17)
F om 4(0.50 g, 0.74 mmol) and BzCl (258
l
L, 2.22 mmol)
acco ding o Gene al p ocedu e II (Sec ion 4.7). Pu ified by column
400
ch oma og aphy (1:2 E OAc/hexane) o yield 400 mg (62%) o 17 as
a yellow amo phous p oduc . [
a
]
D
= –295 (c0.16, CHCl
3
); R
: 0.34
(1:2 E OAc/hexane). ESI-MS (posi i e mode) m/z:890.33 [M+H]
+
.
Anal. Calcd o C
50
H
39
N
3
O
11
S (889.92): C, 67.48, H, 4.42; N, 4.72;
S, 3.60. Found: C, 67.42; H, 4.45; N, 5.71; S, 3.63.
majo isome :
1
H NMR (CDCl
3
, 400 MHz) d(ppm) 9.26 (1H, s,
NH) 8.05–7.10 (30H, m, A ), 6.34 (1H, s, CH- hiadiazoline), 6.02
(1H, p , J= 9.5 Hz, H-2 o H-3 o H-4), 5.72–5.60 (2H, m, H-2 and/
o H-3 and/o H-4), 4.62 (1H, d, J
1,2
= 9.7 Hz, H-1), 4.54 (1H, dd,
J
5,6a
= 5.3 Hz, J
6a,6b
= 12.3 Hz, H-6a), 4.48 (1H, dd, J
5,6b
= 1.9 Hz,
410
H-6b), 4.17–4.11 (1H, m, H-5).
13
C NMR (CDCl
3
, 400 MHz) d
(ppm) 166.6, 166.2, 165.7, 165.7, 165.1, 165.0 (CO, NHCO), 148.4
(Cq- hiadiazoline) 133.7–127.3 (A ), 76.5, 75.9, 73.7, 69.2, 67.7
(C-1 o C-5), 69.1 (CH- hiadiazoline), 62.6 (C-6).
Mino isome :
1
H NMR (CDCl
3
, 400 MHz) d(ppm) 9.10 (1H, s,
NH), 6.63 (1H, s, CH- hiadiazoline), 6.09, 5.90 (2H, 2 p , J= 9.5 Hz
in each, H-2 and/o H-3 and/o H-4), 4.72 (1H, d, J
1,2
= 9.1 Hz,
B. Sz}
ocs e al. / Ca bohyd a e Resea ch xxx (2013) xxx–xxx 5
CAR 6420 No. o Pages 10, Model 5G
27 Ma ch 2013
Please ci e his a icle in p ess as: Sz}
ocs, B.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.03.009
H-1), 4.67 (1H, dd, J
5,6a
= 2.3 Hz, J
6a,6b
= 12.4 Hz, H-6a), 4.41 (1H, dd,
J
5,6b
= 4.9 Hz, H-6b), 4.26–4.20 (1H, m, H-5).
13
C NMR (CDCl
3
,
400 MHz) d(ppm) 76.1, 74.8, 68.6 (C-1 o C-5), 60.3 (C-6).
420
4.7.3. 2-(2-Naph hamido)-4-(2-naph oyl)-5-(2,3,4,6- e a-O-
benzoyl-b-
D
-glucopy anosyl)-
D
2
-1,3,4- hiadiazoline (18)
F om 4(0.50 g, 0.74 mmol) and 2-naph hoyl chlo ide (423 mg,
2.22 mmol) acco ding o Gene al p ocedu e II (Sec ion 4.7). Pu i-
fied by column ch oma og aphy (1:2 E OAc/hexane) o yield
450 mg (62%) o 18 as a yellow amo phous p oduc . [
a
]
D
= –318
(c0.16, CHCl
3
); R
: 0.43 (1:8 E OAc/ oluene). ESI-MS (posi i e
mode) m/z:990.33 [M+H]
+
. Anal. Calcd o C
58
H
43
N
3
O
11
S
(989.26): C, 70.36, H, 4.38; N, 4.24; S, 3.24. Found: C, 70.32; H,
4.36; N, 4.21; S, 3.27.
430
Majo isome :
1
H NMR (CDCl
3
, 400 MHz) d(ppm) 9.30 (1H, s,
NH) 8.36–6.84 (34H, m, A ), 6.36 (1H, d, CH- hiadiazoline), 5.96
(1H, p , J= 9.5 Hz, H-2 o H-3 o H-4), 5.70–5.61 (2H, m, H-2 and/
o H-3 and/o H-4), 4.60 (1H, dd, J
1,CH
= 1.4 Hz, J
1,2
= 9.9 Hz, H-1),
4.52 (1H, dd, J
6a,6b
= 12.2 Hz, H-6a,), 4.46 (1H, dd, H-6b), 3.97
(1H, ddd, J
5,6b
= 2.7 Hz, J
5,6a
= 4.9 Hz, J
4,5
= 9.7 Hz, H-5).
13
C NMR
(CDCl
3
, 400 MHz) d(ppm) 169.7, 166.5, 166.3, 165.8, 165.7, 165.0
(CO, NHCO), 148.1 (Cq- hiadiazoline), 135.3–123.5 (A ), 76.4,
75.8, 73.7, 69.1, 68.0 (C-1 o C-5), 69.1 (CH- hiadiazoline), 62.5
(C-6).
440
Mino isome :
1
H NMR (CDCl
3
, 400 MHz) d(ppm) 6.68 (1H, d,
CH- hiadiazoline), 6.13, 5.89 (2H, 2 p , J= 9.5 Hz in each, H-2
and/o H-3 and/o H-4), 4.76 (1H, dd, J
1,CH
= 2.5 Hz, J
1,2
= 9.8 Hz,
H-1), 4.65 (1H, dd, J
6a,6b
= 12.3 Hz, H-6a), 4.19 (1H, ddd,
J
5,6a
= 2.8 Hz, J
5,6b
= 4.6 Hz, J
4,5
= 9.6 Hz, H-5).
13
C NMR (CDCl
3
,
400 MHz) d(ppm) 167.3, 166.2, 165.4, 164.9 (CO, NHCO), 148.7
(Cq- hiadiazoline), 76.1, 75.4, 74.8, 68.5, 67.8 (C-1 o C-5), 63.0
(C-6).
4.7.4. 2-(4- e -Bu ylbenzamido)-4-(4- e -bu ylbenzoyl)-5-
(2,3,4,6- e a-O-benzoyl-b-
D
-glucopy anosyl)-
D
2
-1,3,4-
450
hiadiazoline (19)
F om 4(0.15 g, 0.22 mmol) and 4- e -bu ylbenzoyl chlo ide
(129
l
L, 0.66 mmol) acco ding o Gene al p ocedu e II (Sec ion
4.7). Pu ified by column ch oma og aphy (1:4 E OAc/hexane) o
yield 83 mg (38%) o 19 as a yellow amo phous p oduc . [
a
]
D
=–
378 (c0.18, CHCl
3
); R
: 0.48 (1:2 E OAc/hexane);
1
H NMR (CDCl
3
,
360 MHz) d(ppm) 8.71 (1H, s, NH), 8.01–7.14 (28H, m, A ), 6.36
(1H, d, J= 1.4 Hz, CH- hiadiazoline), 6.01, 5.65, 5.64 (3H, 3 p ,
J= 9.5 Hz in each, H-2, H-3, H-4), 4.64–4.55 (2H, m, H-6a, H-1),
4.42 (1H, dd, J
6a,6b
= 12.0 Hz, H-6b), 4.14 (1H, ddd, J
5,6b
= 2.0 Hz,
460
J
5,6a
= 5.7 Hz, J
4,5
= 9.5 Hz, H-5), 1.36 (9H, b s, CH
3
), 1.31 (9H, s,
CH
3
).
13
C NMR (CDCl
3
, 360 MHz) d(ppm) 166.4, 166.3, 165.8,
165.8, 165.2, 164.7 (CO, NHCO), 156.9, 154.3 (Cq-phenyl), 145.9
(Cq- hiadiazoline), 133.7–124.7 (A ), 76.6, 76.0, 73.8, 69.3, 67.8,
(C-1 o C-5), 69.1 (CH- hiadiazoline), 62.8 (C-6), 35.2, 34.9 (Cq),
31.2, 31.1 (CH
3
). ESI-MS (posi i e mode) m/z:1002.33 [M+H]
+
.
Anal. Calcd o C
58
H
55
N
3
O
11
S (1002.14): C, 69.51, H, 5.53; N,
4.19; S, 3.20. Found: C, 69.54; H, 5.56; N, 4.16; S, 3.25.
4.8. Gene al p ocedu e III o he syn hesis o 2-acylamino-5-
(2,3,4,6- e a-O-benzoyl-b-
D
-glucopy anosyl)-1,3,4-oxadiazoles
470
and - hiadiazoles
A 4-acyl-[C-(2,3,4,6- e a-O-benzoyl-b-
D
-glucopy anosyl) o m-
aldehyde]semica bazone (12–15, 0.03 mmol) o a 2-acylamino-
4-acyl-5-(2,3,4,6- e a-O-benzoyl-b-
D
-glucopy anosyl)-1,3,4- hi-
adiazoline (16–19, 0.03 mmol) was dissol ed in CH
2
Cl
2
(1 mL).
Then PIDA (1.1 equi ) was added, and he mix u e was s i ed
a oom empe a u e. The eac ion was moni o ed by TLC (1:1
E OAc/hexane). When he eac ion was comple e, he sol en
was e apo a ed and he esidue was pu ified by column
ch oma og aphy.
480
4.8.1. 2-Ace amido-5-(2,3,4,6- e a-O-benzoyl-b-
D
-
glucopy anosyl)-1,3,4-oxadiazole (20)
F om 12 (0.16 g, 0.23 mmol) acco ding o Gene al p ocedu e III
(Sec ion 4.8). Pu ified by column ch oma og aphy (1:3 E OAc/hex-
ane) o yield 81 mg (50%) o 20 as a whi e amo phous p oduc .
[
a
]
D
=–2 (c0.25, CHCl
3
); R
: 0.38 (1:3 E OAc/ oluene);
1
HNMR
(CDCl
3
, 360 MHz) d(ppm) 8.03–7.22 (21H, m, A , NH), 6.08, 5.86,
5.84 (3H, 3 p , J= 9.7 Hz in each H-2, H-3, H-4), 5.18 (1H d,
J
1,2
= 9.8 Hz, H-1), 4.71–4.60 (1H, dd, J
5,6a
< 1 Hz, H-6a), 4.52 (1H,
dd, J
5,6b
= 4.3 Hz, J
6a,6b
= 12.1 Hz, H-6b), 4.40–4.30 (1H, m, H-5),
490
2.53 (1H, s, CH
3
).
13
C NMR (CDCl
3
, 360 MHz) d(ppm) 166.0,
165.6, 165.2, 165.0, 164.8, 160.9 (CO, NHCO, C-oxadiazole),
133.5–128.3 (A ), 76.9, 73.4, 71.8, 70.3, 68.9 (C-1 o C-5), 62.9 (C-
6), 11.0 (CH
3
). Anal. Calcd o C
38
H
31
N
3
O
11
(705.67): C, 64.68, H,
4.43; N, 5.95. Found: C, 64.60; H, 4.51; N, 5.86.
4.8.2. 2-Benzamido-5-(2,3,4,6- e a-O-benzoyl-b-
D
-
glucopy anosyl)-1,3,4-oxadiazole (21)
F om 13 (0.084 g, 0.11 mmol) acco ding o Gene al p ocedu e III
(Sec ion 4.8). Pu ified by column ch oma og aphy (1:3 E OAc/hex-
ane) o yield 70 mg (57%) o 21 as a whi e amo phous p oduc .
500
[
a
]
D
= –205 (c0.4, CHCl
3
); R
: 0.68 (1:1 E OAc/hexane);
1
HNMR
(CDCl
3
, 360 MHz) d(ppm) 8.06–7.26 (26H, m, A , NH), 6.11, 6.02,
5.88 (3H, 3 p , J= 9.5, 9.8 Hz in each H-2, H-3, H-4), 5.28 (1H d,
J
1,2
= 9.8 Hz, H-1), 4.71 (1H, dd, J
6a,6b
= 12.4 Hz, H-6a), 4.54 (1H,
dd, H-6b) 4.39 (1H, ddd, J
5,6a
= 2.4 Hz, J
5,6b
= 5.2 Hz, J
4,5
= 9.4 Hz,
H-5).
13
C NMR (CDCl
3
, 360 MHz) d(ppm) 166.0, 165.9, 165.6,
165.1, 164.8, 160.9 (CO, NHCO, C-oxadiazole), 133.5–123.3 (A ),
77.0, 73.5, 71.8, 70.2, 69.0 (C-1 o C-5), 62.9 (C-6). Anal. Calcd o
C
43
H
33
N
3
O
11
(767.74): C, 67.27, H, 4.33; N, 5.47. Found: C, 67.35;
H, 4.42; N, 5.40.
510
4.8.3. 2-(2-Naph hamido)-5-(2,3,4,6- e a-O-benzoyl-b-
D
-
glucopy anosyl)-1,3,4-oxadiazole (22)
F om 14 (0.28 g, 0.34 mmol) acco ding o Gene al p ocedu e III
(Sec ion 4.8). Pu ified by column ch oma og aphy (1:6 E OAc/ olu-
ene) o yield 192 mg (69%) o 22 as a yellow amo phous p oduc .
[
a
]
D
= –121 (c0.16, CHCl
3
); R
:0.58 (1:3.5 E OAc- oluene);
1
H
NMR (CDCl
3
, 360 MHz) d(ppm) 8.52 (1H, s, NH), 8.11–7.10 (27H,
m, A ), 6.18, 6.11, 5.95 (3H, 3 p , J= 9.7 Hz in each, H-2, H-3,
H-4), 5.27 (1H, d, J
1,2
= 9.5 Hz, H-1), 4.70 (1H, dd, J
6a,6b
= 12.4 Hz,
H-6a), 4.54 (1H, dd, H-6b) 4.39 (1H, ddd, J
5,6a
= 2.7 Hz,
520
J
5,6b
= 4.3 Hz, J
4,5
= 9.5 Hz, H-5).
13
C NMR (CDCl
3
, 360 MHz) d
(ppm) 166.0, 165.9, 165.6, 164.8, 161.1 (CO, NHCO, C-oxadiazole),
134.7–120.5 (A ), 77.0, 73.6, 71.9, 70.3, 69.1 (C-1 o C-5), 63.0
(C-6). Anal. Calcd o C
47
H
35
N
3
O
11
(817.79): C, 69.03, H, 4.31; N,
5.14. Found: C, 69.11; H, 4.41; N, 5.05.
4.8.4. 2-(4- e -Bu ylbenzamido)-5-(2,3,4,6- e a-O-benzoyl-b-
D
-
glucopy anosyl)-1,3,4-oxadiazole (23)
F om 15 (0.09 g, 0.11 mmol) acco ding o Gene al p ocedu e III
(Sec ion 4.8). Pu ified by column ch oma og aphy (1:3 E OAc/hex-
ane) o yield 80 mg (60%) o 23 as a colou less amo phous p oduc .
530
[
a
]
D
= –82 (c1.50, CHCl
3
); R
: 0.29 (1:3 E OAc/hexane);
1
HNMR
(CDCl
3
, 360 MHz) d(ppm) 8.05–7.20 (25H, m, A , NH), 6.11, 6.02,
5.88 (3H, 3 p , J= 9.5, 9.7 Hz in each, H-2, H-3, H-4), 5.28 (1H, d,
J
1,2
= 9.7 Hz, H-1), 4.70 (1H, dd, J
6a,6b
= 12.4 Hz, H-6a), 4.54 (1H,
dd, H-6b), 4.40 (1H, ddd, J
5,6a
= 2.3 Hz, J
5,6b
= 5.3 Hz, J
4,5
= 9.7 Hz,
H-5), 1.34 (9H, s, CH
3
).
13
C NMR (CDCl
3
, 360 MHz) d(ppm) 166.0,
165.9, 165.6, 165.1, 164.8, 161.1 (CO, NHCO, C-oxadiazole), 155.6,
133.5–120.5 (A ), 77.0, 73.6, 71.8, 70.2, 69.1 (C-1 o C-5), 63.0
(C-6), 35.0 (Cq- Bu), 31.0 (CH
3
). Anal. Calcd o C
47
H
41
N
3
O
11
6B. Sz}
ocs e al. / Ca bohyd a e Resea ch xxx (2013) xxx–xxx
CAR 6420 No. o Pages 10, Model 5G
27 Ma ch 2013
Please ci e his a icle in p ess as: Sz}
ocs, B.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.03.009
(823.84): C, 68.52, H, 5.02; N, 5.10. Found: C, 68.61; H, 5.11; N,
540
5.18.
4.8.5. 2-Ace amido-5-(2,3,4,6- e a-O-benzoyl-b-
D
-
glucopy anosyl)-1,3,4- hiadiazole (24)
F om 16 (0.17 g, 0.22 mmol) acco ding o Gene al p ocedu e III
(Sec ion 4.8). Pu ified by column ch oma og aphy (1:2 E OAc/ olu-
ene) o yield 106 mg (67%) o 24 as a whi e amo phous p oduc .
[
a
]
D
= –81 (c0.25, CHCl
3
); R
: 0.42 (1:2 E OAc/ oluene);
1
H NMR
(CDCl
3
, 360 MHz) d(ppm) 8.09–7.13 (21H, m, A , NH), 6.09, 5.81,
5.74 (3H, 3 p , J= 9.6, 9.8 Hz in each, H-2, H-3, H-4), 5.25 (1H, d,
J
1,2
= 9.6 Hz, H-1), 4.70 (1H, dd, J
6a,6b
= 12.3 Hz, H-6a), 4.53 (1H,
550
dd, H-6b), 4.36 (1H, ddd, J
5,6a
= 3.0 Hz, J
5,6b
= 4.8 Hz, J
4,5
= 9.5 Hz,
H-5), 1.82 (3H, s, CH
3
).
13
C NMR (CDCl
3
, 360 MHz) d(ppm) 168.8,
166.1, 165.7, 165.1, 164.7 (CO, NHCO, C- hiadiazole), 133.5–128.4
(A ), 76.9, 76.0, 73.5, 71.7, 69.2 (C-1 o C-5), 62.9 (C-6). ESI-MS (po-
si i e mode) m/z:722.33 [M+H]
+
. Anal. Calcd o C
38
H
31
N
3
O
10
S
(721.76): C, 63.24, H, 4.33; N, 5.82; S,4.44. Found: C, 63.21; H,
4.35; N, 5.84; S, 4.41.
4.8.6. 2-Benzamido-5-(2,3,4,6- e a-O-benzoyl-b-
D
-
glucopy anosyl)-1,3,4- hiadiazole (25)
F om 17 (0.09 g, 0.10 mmol) acco ding o Gene al p ocedu e III
560
(Sec ion 4.8). Pu ified by column ch oma og aphy (1:3 E OAc/hex-
ane) o yield 54 mg (68%) o 25 as a whi e amo phous p oduc .
[
a
]
D
= –111 (c0.20, CHCl
3
); R
: 0.21 (1:2 E OAc/hexane);
1
H NMR
(CDCl
3
, 360 MHz) d(ppm) 8.20–7.26 (26H, m, A , NH), 6.12, 5.86,
5.77 (3H, 3 p , J= 9.5, 9.8 Hz in each, H-2, H-3, H-4), 5.32 (1H, d,
J
1,2
= 9.7 Hz, H-1), 4.70 (1H, dd, J
6a,6b
= 12.4 Hz, H-6a), 4.55 (1H,
dd, H-6b) 4.40 (1H, ddd, J
5,6a
= 2.7 Hz, J
5,6b
= 5.0 Hz, J
4,5
= 9.7 Hz,
H-5).
13
C NMR (CDCl
3
, 360 MHz) d(ppm) 166.2, 165.7, 165.1,
165.0, 164.9, 162.5, 161.1 (CO, NHCO, C- hiadiazole), 133.3–128.3
(A ), 77.0, 76.3, 73.6, 72.2, 69.2, (C-1 o C-5), 63.1 (C-6). Anal. Calcd
570
o C
43
H
33
N
3
O
10
S (783.8): C, 65.89, H, 4.24; N, 5.36; S,4.09. Found:
C, 65.94; H, 4.35; N, 5.28; S, 4.01.
4.8.7. 2-(2-Naph hamido)-5-(2,3,4,6- e a-O-benzoyl-b-
D
-
glucopy anosyl)-1,3,4- hiadiazole (26)
F om 18 (0.20 g, 0.20 mmol) acco ding o Gene al p ocedu e III
(Sec ion 4.8). Pu ified by column ch oma og aphy (1:6 E OAc/ olu-
ene) o yield 100 mg (59%) o 26 as a yellow amo phous p oduc .
[
a
]
D
=205 (c0.16, CHCl
3
); R
: 0.39 (1:6 E OAc/ oluene);
1
H NMR
(CDCl
3
, 360 MHz) d(ppm) 8.71 (s, 1H, NH), 8.12–7.11 (27H, m,
A ), 6.15, 5.91, 5.80 (3H, 3p , J= 9.6 Hz in each, H-2, H-3, H-4),
580
5.35 (1H, d, J
1,2
= 9.7 Hz, H-1), 4.70 (1H, dd, J
ia,6b
= 12.3 Hz, H-6a),
4.57 (1H, dd, H-6b), 4.41 (1H, ddd, J
5,6a
= 2.0 Hz, J
5,6b
= 4.9 Hz,
J
4,5
= 9.4 Hz, H-5).
13
C NMR (CDCl
3
, 360 MHz) d(ppm) 166.1,
165.7, 165.2, 165.1, 165.0, 162.5, 160.9 (CO, NHCO, C- hiadiazole),
135.5–124.2 (A ), 76.9, 76.3, 73.6, 72.3, 69.3 (C-1 o C-5), 63.1 (C-
6). ESI-MS (posi i e mode) m/z:834.33 [M+H]
+
. Anal. Calcd o
C
47
H
35
N
3
O
10
S (833.86): C, 67.70, H, 4.23; N, 5.04; S, 3.85. Found:
C, 67.81; H, 4.15; N, 5.12; S, 3.76.
4.8.8. 2-(4- e -Bu ylbenzamido)-5-(2,3,4,6- e a-O-benzoyl-b-
D
-
glucopy anosyl)-1,3,4- hiadiazole (27)
590
F om 19 (0.17 g, 0.17 mmol) acco ding o Gene al p ocedu e III
(Sec ion 4.8). Pu ified by column ch oma og aphy (1:3 E OAc/hex-
ane) o yield 120 mg (84%) o 27 as a whi e amo phous p oduc .
[
a
]
D
= –179 (c0.20, CHCl
3
); R
: 0.20 (1:3 E OAc/hexane);
1
H NMR
(CDCl
3
, 360 MHz) d(ppm) 8.05–7.22 (25H, m, A , NH), 6.12, 5.86,
5.80 (3H, 3 p , J= 9.5, 9.7 Hz in each, H-2, H-3, H-4), 5.31 (1H, d,
J
1,2
= 9.6 Hz, H-1), 4.68 (1H, dd, J
5,6a
< 1 Hz, J
6a,6b
= 12.2 Hz, H-6a),
4.54 (1H, dd, J
5,6b
= 5.0 Hz, H-6b), 4.43–4.35 (1H, m, H-5), 1.38
(9H, s, CH
3
).
13
C NMR (CDCl
3
, 360 MHz) d(ppm) 166.1, 165.8,
165.2, 165.0, 164.8, 161.8, 160.9 (CO, NHCO, C- hiadiazole),
600
156.9, 133.5–125.8 (A ), 76.9, 76.3, 73.7, 72.2, 69.4 (C-1 o C-5),
63.2 (C-6), 31.9 (Cq- Bu), 31.1 (CH
3
). ESI-MS (posi i e mode) m/z:
840.42 [M+H]
+
. Anal. Calcd o C
47
H
41
N
3
O
10
S (839.91): C, 67.21,
H, 4.92; N, 5.00; S, 3.82. Found: C, 67.31; H, 4.83; N, 5.09; S, 3.91.
4.9. P epa a ion o he es compounds
4.9.1. Gene al p ocedu e IV o he Zemplén debenzoyla ion
An O-pe benzoyla ed compound (100 mg) was dissol ed in d y
MeOH (1 mL) and a solu ion o NaOMe (1 M in MeOH) was added
o he solu ion in a ca aly ic amoun . The eac ion mix u e was
kep a . When he eac ion was comple e (TLC, 7:3 CHCl
3
/MeOH)
610
he solu ion was neu alized wi h a ca ion exchange esin Ambe -
lys 15 (H
+
o m). The esin was fil e ed o wi h suc ion, he fil a e
was e apo a ed unde educed p essu e. The c ude p oduc was
pu ified by column ch oma og aphy.
4.9.2. Gene al p ocedu e V o he emo al o O-benzoyl
p o ec ing g oups
An O-pe benzoyla ed compound (100 mg) was dissol ed in d y
MeOH (14 mL) and LiOH (16 equi ) was added. The eac ion mix-
u e was s i ed a 0 °C. When he eac ion was comple e (TLC, 7:3
CHCl
3
/MeOH) he solu ion was neu alized wi h a ca ion exchange
620
esin Ambe lys 15 (H
+
o m). A e fil a ion and emo al o he sol-
en , he esidue was pu ified by column ch oma og aphy.
4.9.3. C-(b-
D
-Glucopy anosyl) o maldehyde semica bazone (6)
F om 2(0.17 g, 0.26 mmol) acco ding o Gene al p ocedu e IV
(Sec ion 4.9.1). Pu ified by column ch oma og aphy (8:5 CHCl
3
/
me hanol) o yield 40 mg (63%) o 6as a whi e amo phous p oduc .
[
a
]
D
= +32 (c0.30, DMSO); R
: 0.17 (2:1 CHCl
3
/me hanol);
1
HNMR
(MeOD, 360 MHz) d(ppm) 7.19 (1H, s, CH@N), 3.88–3.72 (2H, m,
H-1, H-2 o H-3 o H-4), 3.70–3.52 (1H, m, H-6a), 3.50–3.22 (4H,
m, H-2 and/o H-3 and/o H-4, H-5, H-6b).
13
C NMR (MeOD,
630
360 MHz) d(ppm) 160.1 (NHCONH), 143.2 (CH@N), 81.8, 80.2,
79.4, 73.4, 71.4 (C-1 o C-5), 62.7 (C-6). Anal. Calcd o
C
84
H
15
N
3
O
6
(249.22): C, 38.55, H, 6.07; N, 16.86. Found: C, 38.63;
H, 6.12; N, 16.93.
4.9.4. 4-Phenyl-[C-(b-
D
-glucopy anosyl) o maldehyde]
semica bazone (7)
F om 3(0.20 g, 0.27 mmol) acco ding o Gene al p ocedu e IV
(Sec ion 4.9.1). Pu ified by column ch oma og aphy (4:1 CHCl
3
/
me hanol) o yield 42 mg (48%) o 7as a whi e amo phous p oduc .
[
a
]
D
= +52 (c0.43, DMSO); R
: 0.23 (4:1 CHCl
3
/me hanol);
1
HNMR
640
(MeOD, 360 MHz) d(ppm) 7.46 (2H, d, J= 7.9 Hz, A ), 7.30–7.17
(3H, m, A , CH@N), 7.01 (1H, , J= 7.3 Hz, A ), 3.91–3.79 (2H, m,
H-1, H-2 o H-3 o H-4), 3.65 (1H, dd, J
5,6a
= 4.5 Hz, J
6a,6b
= 11.4 Hz,
H-6a), 3.42–3.21 (4H, m, H-2 and/o H-3 and/o H-4, H-5, H-6b),
13
C NMR (MeOD, 360 MHz) d(ppm) 155.9 (NHCONH), 143.3
(CH@N), 139.7, 129.8, 124.5, 121.3 (A ), 82.0, 80.2, 79.5, 73.5,
71.5 (C-1/C-5), 62.8 (C-6). Anal. Calcd o C
14
H
19
N
3
O
6
(325.32): C,
51.69, H, 5.89; N, 12.92. Found: C, 51.79; H, 5.98; N, 12.85.
4.9.5. C-(b-
D
-glucopy anosyl) o maldehyde hiosemica bazone
(8)
650
F om 4(0.16 g, 0.24 mmol) acco ding o Gene al p ocedu e V
(Sec ion 4.9.2). Pu ified by column ch oma og aphy (3:1 CHCl
3
/
me hanol) o yield 60 mg (95%) o 8as a whi e amo phous p oduc .
[
a
]
D
=+6(c2.00, DMSO); R
: 0.24 (3:1 CHCl
3
/me hanol);
1
HNMR
(MeOD, 360 MHz) d(ppm) 7.39 (1H, s, CH@N), 3.92–3.81 (2H, m,
H-1, H-2 o H-3 o H-4), 3.68 (1H, dd, J
5,6a
= 3.1 Hz, J
6a,6b
= 10.9 Hz,
H-6a), 3.52–3.32 (4H, m, H-2 and/o H-3 and/o H-4, H-5, H-6b).
13
C NMR (MeOD, 360 MHz) d(ppm) 180.0 (NHCSNH), 145.5
(CH@N), 81.7, 79.8, 79.0, 73.2, 71.4 (C-1/C-5), 62.7 (C-6). Anal.
Calcd o C
8
H
15
N
3
O
5
S (265.29): C, 36.22, H, 5.70; N, 15.84; S,
660
12.09. Found: C, 36.12; H, 5.81; N, 15.92; S, 12.17.
B. Sz}
ocs e al. / Ca bohyd a e Resea ch xxx (2013) xxx–xxx 7
CAR 6420 No. o Pages 10, Model 5G
27 Ma ch 2013
Please ci e his a icle in p ess as: Sz}
ocs, B.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.03.009