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G aphical abs ac
pp xxx–xxxSyn hesis o 2-(b-
DD
-glucopy anosylamino)-5-subs i u ed-1,3,4-oxadiazoles o inhibi ion o glycogen
phospho ylase
Ma ie a Tó h
*
, Béla Sz}
ocs, Tímea Kaszás, Tibo Docsa, Pál Ge gely, László Somsák
*
O
OAc
AcO
AcO
OAc
NCO
O
OAc
AcO
A
cO
OAc
H
N
O
H
NNA
O
OH
HO
HO
OH
H
N
O
H
NNA
A = Ph, 4-Me-Ph, 4-NO
2
-Ph, 4-F-Ph,
3-Cl-Ph, 1-Naph hyl, 2-Naph hyl
O
OH
HO
HO
OH
H
N
NN
OA
Bes inhibi o :
A = 4-NO
2
-Ph (K
i
=4.5µM)
Bes inhibi o :
A = 4-Me-Ph (K
i
=12µM)
CAR 6468 No. o Pages 1, Model 5G
1 May 2013
Highligh s
P epa a ion o a oma ic aldehyde 4-(b-
DD
-glucopy anosyl)semica bazones. Syn hesis o 2-(b-
DD
-glucopy anosylamino)-5-subs i u ed-1,3,4-
oxadiazoles. Low mic omola inhibi o s o glycogen phospho ylase.
1
Syn hesis o 2-(b-D-glucopy anosylamino)-5-subs i u ed-1,3,4-oxadi-
azoles o inhibi ion o glycogen phospho ylase
Ma ie a Tó h
a,
⇑
,Béla Sz}
ocs
a
,Tímea Kaszás
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 13 Ma ch 2013
Recei ed in e ised o m 18 Ap il 2013
Accep ed 18 Ap il 2013
A ailable online xxxx
20 Keywo ds:
Semica bazone
1,3,4-Oxadiazole
b-
D
-Glucopy anosyl de i a i es
Glycogen phospho ylase
Inhibi o
abs ac
A oma ic aldehyde 4-(2,3,4,6- e a-O-ace yl-b-D-glucopy anosyl)semica bazones we e syn hesized by
he addi ion o di e en hyd azones on o O-pe ace yla ed b-D-glucopy anosyl isocyana e. Oxida i e
ans o ma ions o hese p ecu so s ga e O-p o ec ed 2-(b-D-glucopy anosylamino)-5-subs i u ed-
1,3,4-oxadiazoles. Remo al o he O-ace yl p o ec ing g oups unde Zemplén condi ions ga e es
compounds o show low mic omola inhibi ion agains abbi muscle glycogen phospho ylase b. Bes
inhibi o s o hese se ies we e 4-(b-D-glucopy anosyl)semica bazones o 4-fluo obenzaldehyde
(K
i
= 4.5
l
M), 2-naph haldehyde (K
i
= 5.5
l
M) and 2-(b-D-glucopy anosylamino)-5-(4-me hylphenyl)-
1,3,4-oxadiazole (K
i
=12
l
M).
Ó2013 Published by Else ie L d.
1. In oduc ion
40
Inhibi ion o glycogen phospho ylase (GP) can be a new
he apeu ic me hod o he ea men o ype 2 diabe es melli us,
and po en ial applica ion o GP inhibi o s (GPIs) in some o he
diseased s a es, such as ea ly ca diac and ca dio ascula diso de s
in non-diabe ics, ca diac a hy hmias, ischemic inju ies, and umo
g ow h was also p oposed.
1–5
A la ge a ay o compounds was
shown o ha e inhibi o y e ec agains GP unde in i o condi-
ions.
4,6
Among hese molecules glucose de i a i es a e he mos
in ensi ely in es iga ed GPIs, and de ailed s udies can be ound
in he li e a u e on hei s uc u e–ac i i y ela ionships (SAR).
5,7
50
Some glucose de i ed GPIs had in i o hypoglycaemic
8
and o he
in e es ing physiological e ec s.
9
Widely s udied glucose analogue GPIs a e de i a i es o
b-
D
-glucopy anosylamine, such as N-acyl-b-
D
-glucopy anosylam-
ines
10–13
(Iin Cha 1,e.g., K
i
=10–13
l
M
11,14
agains abbi muscle
GPb(RMGPb)
15
o R = 2-naph hyl), N-a yl-N
0
-b-
D
-glucopy anosyl
u eas
4
(II,e.g., K
i
= 5.2
l
M (RMGPb) o R = 2-naph hyl), as well
as N-acyl-N
0
-b-
D
-glucopy anosyl u ea de i a i es
4,16
(III,e.g.,
K
i
= 0.35
l
M (RMGPb) o R = 2-naph hyl) which inhibi ed he en-
zyme in o below he low mic omola ange. Mic omola e ficiency
60
was epo ed also o 4-(b-
D
-glucopy anosyl) hiosemica bazones
o a oma ic aldehydes
17,18
(IV X=S, e.g., IC
50
= 5.7
l
M (RMGPb)
o A = 4-fluo ophenyl). Compounds V, in which he suga and
he a oma ic pa s a e in e changed in compa ison o IV, ha e e y
ecen ly been shown o be GPIs (K
i
=29
l
M o VX = O, and
K
i
= 300
l
M o VX = S agains RMGPb).
19
Replacemen o he NHCO moie y by 1,2,3- iazole (a non-
classical bioisos e ic he e ocyclic linke A) in molecules I esul ed
in e ec i e GP inhibi o s
20
(IA,e.g., K
i
=16
l
M (RMGPb) o
R = 2-naph hyl). Enzyma ic es s and c ys allog aphic s udies ha e
70
shown high simila i y o he amide (I) and he 1,2,3- iazole (IA)
ype molecules bo h in binding s eng h and s uc u al ea u es
o he enzyme–inhibi o complexes.
14
Replacemen o he NHCO
moie y wi h isome ic oxadiazoles B,C, and D esul ed in inhibi o s
IB–D wi h a ying e ficiency. Among hese molecules he 3-a yl-5-
b-
D
-glucopy anosyl-1,2,4-oxadiazoles IC p o ed o be he mos
e ec i e compounds (e.g., K
i
= 2.4
l
M(RMGPb) o R = 2-naph-
hyl).
21,22
Bioisos e ic eplacemen s udies we e also ca ied ou
wi h compounds III.
13,19,23
As pa o a p og am o sys ema ically eplace NHCO moie ies o
80
GPIs I–III by he e ocyclic bioisos e es, he ein we epo on he
syn heses and enzyma ic es s o a se ies o 2-(b-
D
-glucopy ano-
sylamino)-5-subs i u ed-1,3,4-oxadiazoles IIB.
Fo he cons uc ion o 1,3,4-oxadiazole ings om acyclic p e-
cu so s acidic ea men o N,N
0
-diacyl- (o N-acyl-N
0
- hioacyl)-
hyd azines as well as oxida i e cycliza ion o acylhyd azones a e
he mos equen ly used p ocedu es, and bo h me hods we e
applied o he syn heses o 2-amino-5-subs i u ed-1,3,4-oxadiaz-
oles, oo.
24
2-Glycosylamino-5-subs i u ed-1,3,4-oxadiazoles a e
also known and ha e been syn hesized om glycosyl iso hiocya-
90
na es ei he by he addi ion o acid hyd azides ollowed by ing
0008-6215/$ - see on ma e Ó2013 Published by Else ie L d.
h p://dx.doi.o g/10.1016/j.ca es.2013.04.025
⇑
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
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 6468 No. o Pages 10, Model 5G
2 May 2013
Please ci e his a icle in p ess as: Tó h, M.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.04.025
closu e o he esul ing 1-acyl-4-glycosyl- hiosemica bazides
25–27
o by chlo ina ion o gi e in e media e glycosyl isocyanide dichlo-
ides o be cyclized wi h acid hyd azides.
28
In his wo k we in es iga ed he syn hesis o compounds IIB
om aldehyde 4-glycosyl-semica bazones (c . O-p o ec ed IV
X=O in Cha 1), a ou e which, o he bes o ou knowledge,
has no ye been applied o ob ain 2-glycosylamino-1,3,4-oxadiaz-
oles. On he o he hand, he p epa a ion o semica bazones IV
(X = O) ga e he oppo uni y o compa e hei GPI p ope ies o
100
he co esponding hiosemica bazones IV (X = S).
2. Resul s and discussion
2.1. Syn heses
Fo he o ma ion o he a ge oxadiazoles oxida i e ing
closu e o a oma ic aldehyde 4-(2,3,4,6- e a-O-ace yl-b-
D
-gluco-
py anosyl)semica bazones (simila o ha applied o he
syn heses o 2-a yl-5-(2,3,4,6- e a-O-benzoyl-b-
D
-glucopy ano-
syl)-1,3,4-oxadiazoles
22
) was en isaged as he key s ep. To his
end, O-pe ace yla ed 4-(b-
D
-glucopy anosyl)semica bazide 2was
p epa ed (Scheme 1) by he eac ion o glucopy anosyl isocyana e
110
1
29
(ob ained om he co esponding glucopy anosylamine
30
)
wi h hyd azine eagen s unde di e en eac ion condi ions
( eagen s: NH
2
NH
2
HCl, NH
2
NH
2
HOAc; sol en s: d y py idine,
d y CH
2
Cl
2
,and 1 equi E
3
N). The bes yield (75%) was achie ed
wi h NH
2
NH
2
HOAc in d y CH
2
Cl
2
in he p esence o E
3
N. In each
o he abo e eac ions bis-glucopy anosyl u ea was also isola ed in
a ious amoun s which could be due o he p esence o aces o
wa e in he mix u es.
31
In o de o ge semica bazones 3–9 condensa ion o semica b-
azide 2wi h he p ope aldehydes was planned. Howe e ,
120
eac ions o 2wi h 2-nap haldehyde in d y E OH o d y oluene
in he p esence o ca aly ic amoun s o ei he AcOH o CF
3
COOH
a eflux empe a u e esul ed in complex eac ion mix u es.
The e o e, b-
D
-glucopy anosyl isocyana e 2was eac ed wi h alde-
hyde hyd azones
32
in boiling d y dioxane o esul in he a ge
molecules 3–9 in good yields. Debenzoyla ions we e pe o med
by he Zemplén p o ocol o gi e excellen yields o semica bazones
10–16.
Ring closing eac ions o he p ecu so s unde oxida i e condi-
ions we e s udied nex . Semica bazones 3,5–7, and 9we e
130
eac ed wi h phenyliodonium diace a e (PIDA) in CH
2
Cl
2
a
(Scheme 2) o u nish he co esponding O-pe ace yla ed 2-(b-
D
-
glucopy anosylamino)-5-subs i u ed-1,3,4-oxadiazoles 17,19–21,
and 23, espec i ely, in mode a e yields. Applica ion o Pb(OAc)
4
as he oxidizing agen in glacial AcOH a 80 °C esul ed in be e
yields o 1,3,4-oxadiazoles 17–22 (compa e yields unde condi-
ions aand bin Scheme 2). Debenzoyla ions we e pe o med by
he Zemplén p o ocol o gi e mos ly good yields o 1,3,4-oxadiaz-
oles 24–30 (Scheme 2).
S uc u al elucida ion o he new compounds was based on NMR
140
spec a. The b-
D
-anome ic configu a ion was indica ed by he 8.1–
9.9 Hz coupling cons an s be ween 1-H and 2-H p o ons in he
4
C
1
con o ma ion o each compound. Coupling o 1-H and N(4)H
(9.2–9.8 Hz) could be obse ed in he spec a o semica bazones
3–9 only. Fo he oxadiazoles 17–23 1-H and NH appea ed as
double s and single s, espec i ely, p obably because o a specific
dihed al angle be ween hese p o ons esul ing in a e y small
coupling cons an . A
1
H spec um o semica bazone 5 eco ded
immedia ely (5 min) a e dissolu ion showed he p esence o
one compound only. A NOE di e ence spec um ob ained by i adi-
150
a ion o he N(2)H signal ga e posi i e NOE o he signal o CH@N
(7.86 ppm) and N(4)H (7.25 ppm) indica ing Econfigu a ion o he
C@Ndouble bond. A e 72 h he
1
H spec um o 5exhibi ed wo
se s o signals as a esul o E/Z isome isa ion (1:1 a io, cha ac e -
is ic chemical shi s (d,ppm) o he Eisome : 10.10 (N(2)H), 7.86
(CH@N), 7.25 (N(4)H); o he Zisome : 8.62 (N(2)H), 7.41 (CH@N),
O
HO
HO
HO
OH H
NH
N
X
IV
N
O
HO
HO
HO
OH H
NR
O
linke
I
O
HO
HO
HO
OH H
NH
N
O
linke
II
R
linke s
N
N
NN
O
NN
N
OO
N
N
ABC
O
HO
HO
HO
OH H
NH
N
O O
R
III
D
X = O, S
O
HO
HO
HO
OH H
CN
H
N
V
H
N
X
H
CA Ph
Cha 1.
O
OAc
AcO
AcO
OAc
NCO
O
OAc
AcO
A
cO
OAc
H
N
O
H
NNH
2
O
OAc
AcO
AcO
O
Ac
H
N
O
H
NNA
2 (75 %)
b
1
O
OH
HO
HO
O
H
H
N
O
H
NNA
c
Ph
4-Me-Ph
4-NO
2
-Ph
4-F-Ph
3-Cl-Ph
1-Naph hyl
2-Naph hyl
A
3 (83%)
4 (72%)
5 (66%)
6 (76%)
7 (71%)
8 (89%)
9 (80%)
10 (99%)
11 (98%)
12 (93%)
13 (98%)
14 (94%)
15 (80%)
16 (98%)
61-019-3
a
Scheme 1. Reagen s and condi ions: (a) NH
2
NH
2
HOAc, d y CH
2
Cl
2
,E
3
N, ; (b) A CH@N–NH
2
, d y dioxane, eflux; (c) NaOMe, d y MeOH, .
2M. Tó h e al. / Ca bohyd a e Resea ch xxx (2013) xxx–xxx
CAR 6468 No. o Pages 10, Model 5G
2 May 2013
Please ci e his a icle in p ess as: Tó h, M.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.04.025
7.15 (N(4)H). These upfield shi s o he Zisome a e in acco d wi h
li e a u e expe iences o semica bazones,
33
howe e , con as
hose o O-pe ace yla ed 4-(b-
D
-glucopy anosyl) hiosemica ba-
zones whe e he N(2)Hsignals we e epo ed o appea a 10.1
160
(E) and 14.6 (Z) ppm.
17
2.2. Enzyme inhibi ion s udies
The kine ic pa ame e s (inhibi ion po ency agains abbi mus-
cle glycogen phospho ylase b(RMGPb)) o he dep o ec ed com-
pounds we e de e mined acco ding o he p o ocol desc ibed
ea lie .
34
The esul s a e summa ized in Table 1 showing he inhib-
i o y e ficiency o some ele an e e ence compounds, as well.
Type IV semica bazones 10–16, he ‘open chain’ p ecu so s o
he a ge compounds o his wo k, p o ed low mic omola inhib-
i o s wi h he p ac ically equipo en 4-ni ophenyl (12) and
170
2-naph hyl (16) compounds as he bes ones. The s ong binding
o 16 fi s he gene al end obse ed wi h se e al glucose de i a-
i es exhibi ing mos e ficien inhibi ion wi h la ge hyd ophobic
g oups in he aglycons.
4
A compa ison o he s uc u ally ela ed
hiosemica bazone coun e pa s would be possible o he pai s
12–31,13–32, and 14–33, howe e , he epo ed inhibi ion da a
18
o he sul u con aining compounds 31–33 a e IC
50
alues no
di ec ly compa able wi h he inhibi o cons an s. Ne e heless, i
is in e es ing o no e ha he endency is di e en in he wo se -
ies: wi hin semica bazones he 4-ni ophenyl (12) compound
180
while wi hin he hiosemica bazones he 4-fluo ophenyl (32)
de i a i e p o ed 4 o6 imes s onge inhibi o s han he nex
bes ones 14 and 33, espec i ely.
A compa ison o inhibi ion o 10 o ha o he biu e ype com-
pound 37,
4
ha ing he same numbe o a oms be ween he suga
and he a oma ic ing, indica es he la e o be a  wice be e
inhibi o han he o me one. This may unde line he impo ance
o bo h ca bonyl uni s in he binding. On he o he hand, 10 binds
15- old s onge han 38
4
wi h wo C@Omoie ies bu also ha ing
a o a able CH
2
uni in place o NH, he eby emphasizing ha he
190
igidi y o 10 makes an impo an con ibu ion o he binding.
A u he compa ison o 10 o ype Vcompound 39,
19
whe ein
he semica bazone linke is o mally e e sed be ween he
ca bohyd a e ing and he phenyl g oup, shows a mode a e
s eng hening o he inhibi ion ha may e e o he highe
con ibu ion o he second ca bonyl uni o he binding. This is co -
obo a ed by he smalle dec ease caused by he change 37?39
han ha o 37?10. A o mal e e sal o he hiosemica bazone
linke as in 40
19
makes a e y la ge dec ease in he binding
s eng h mos p obably because he hioca bonyl in he posi ion
200
o he ‘second amide’ moie y is much less sui able o make s ong
in e ac ions o he enzyme.
Replacemen o he NHCO moie y in compounds ype II by he
1,3,4-oxadiazole ing (compounds ype IIB:24–30) esul ed in
mode a e GPIs. Fo he phenyl and 1-naph hyl subs i u ed pai s
34–24 and 35–29 he e is no significan change in he binding
s eng h. Subs i u ion in he 4-posi ion o he a oma ic ing (25–
27) esul ed in a small and unspecific inc ease o he e ficiency,
while he 3-chlo o eplacemen (28) made a somewha weake
inhibi o . Based on hese esul s one could a gue ha he 1,3,4-oxa-
210
diazole is an accep able bioisos e ic eplacemen o he NHCO moi-
e y in his sys em. Howe e , his a gumen is weakened by he 5
imes wo se inhibi ion o 30 as compa ed o 36 o which a specu-
la i e explana ion can be ha he la ge and igid aglycon is unable
o find an accomoda able posi ion in he binding si e o he enzyme.
3. Conclusion
Reac ion o O-pe ace yla ed b-
D
-glucopy anosyl isocyana e
wi h di e en hyd azones ga e he co esponding aldehyde 4-(2,
3,4,6- e a-O-ace yl-b-
D
-glucopy anosyl)semica bazones in good
yields. Oxida ion o hese compounds by PIDA o mo e ad an a-
220
geously by Pb(OAc)
4
esul ed in 2-(b-
D
-glucopy anosylamino)-5-
subs i u ed-1,3,4-oxadiazoles. Zemplén deace yla ion u nished
es compounds which p o ed low mic omola inhibi o s agains
RMGPb.S uc u e–ac i i y conside a ions allow o es ima e he
ela i e con ibu ion o he binding by ca bonyl and hioca bonyl
g oups o N-acyl-N
0
-glucopy anosyl u ea ype GPIs, and aise he
possibili y o applying 1,3,4-oxadiazoles as bioisos e ic eplace-
men s o NHCO moie ies.
4. Expe imen al
4.1. Gene al me hods
230
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
(
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. ESIMS we e eco ded
wi h a B uke mic OTOF-Q ins umen . TLC was pe o med on DC-
Alu olle Kieselgel 60 F
254
(Me ck). TLC pla es we e isualized unde
240
UV ligh , and by gen le hea ing. 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). Alde-
hyde hyd azones we e ob ained om he co esponding
aldehydes and H
2
NNH
2
H
2
O acco ding o a li e a u e p ocedu e.
32
4.2. Gene al p ocedu e I o he syn hesis o a oma ic aldehyde
4-(2,3,4,6- e a-O-ace yl-b-
D
-glucopy anosyl)semica bazones
(3–9)
2,3,4,6-Te a-O-ace yl-b-
D
-glucopy anosyl isocyana e
29
(1,
250
0.22 g, 0.59 mmol) was added o he solu ion o an aldehyde hyd a-
zone (0.27 g, 1.80 mmol) in d y dioxane (5 mL). The eac ion mix-
u e was s i ed a eflux empe a u e o 24 h. When he eac ion
O
OR
RO
RO
OR
H
N
NN
OA
O
OAc
AcO
AcO
O
Ac
H
N
O
H
NNA
a
o
b
3-9
c
Ph
4-Me-Ph
4-NO
2
-Ph
4-F-Ph
3-Cl-Ph
1-Naph hyl
2-Naph hyl
A
17 (45%)
18 -
19 (43%)
20 (24%)
21 (52%)
22 -
23 (49%)
24 (70%)
25 (33%)
26 (79%)
27 (83%)
28 (74%)
29 (84%)
30 (84%)
17-23 (R = Ac)
ab
(85%)
(66%)
(53%)
(88%)
(64%)
(89%)
-
24-30 (R = H)
3
4
5
6
7
8
9
Scheme 2. Reagen s and condi ions: (a) PIDA, CH
2
Cl
2,
; (b) Pb(OAc)
4
, AcOH, 80 °C;
(c) NaOMe, d y MeOH, .
M. Tó h e al. / Ca bohyd a e Resea ch xxx (2013) xxx–xxx 3
CAR 6468 No. o Pages 10, Model 5G
2 May 2013
Please ci e his a icle in p ess as: Tó h, M.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.04.025

was comple e (TLC 1:1 E OAc/hexane) he eac ion mix u e was
fil e ed wi h suc ion, and he sol en was e apo a ed unde
educed p essu e. The c ude p oduc was pu ified by column
ch oma og aphy.
4.3. Gene al p ocedu e II o he syn hesis o 5-a yl-2-(2,3,4,6-
e a-O-ace yl-b-
D
-glucopy anosylamino)-1,3,4-oxadiazoles
(17–23)
260
An aldehyde 4-(2,3,4,6- e a-O-ace yl-b-
D
-glucopy anosyl)
semica bazone (3–9, 0.03 mmol/mL) was dissol ed in CH
2
Cl
2
, hen
PIDA (1.1 equi ) was added, and he mix u e was s i ed a . When
he eac ion was comple e (TLC, 1:1 E OAc/hexane) he sol en was
e apo a ed unde educed p essu e, and he esidue was pu ified by
column ch oma og aphy.
4.4. Gene al p ocedu e III o he syn hesis o 5-a yl-2-(2,3,4,6-
e a-O-ace yl-b-
D
-glucopy anosylamino)-1,3,4-oxadiazoles
(17–23)
An aldehyde 4-(2,3,4,6- e a-O-ace yl-b-
D
-glucopy ano-
270
syl)semica bazone (3–9, 0.03 mmol/mL) was dissol ed in glacial
AcOH, hen Pb(OAc)
4
(1 equi ) was added, and he mix u e was
s i ed a 80 °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 (10 mL), and washed wi h E OAc (3 
6 mL). The o ganic laye was sepa a ed, d ied, and he sol en
was e apo a ed unde educed p essu e. The esidue was pu ified
by column ch oma og aphy.
4.5. Gene al p ocedu e IV o he emo al o O-ace yl p o ec ing
g oups
280
An O-pe ace yla ed compound (100 mg) was dissol ed in d y
MeOH (1 mL) and a solu ion o NaOMe (0.1 M in MeOH) was added
o he solu ion in a ca aly ic amoun . The eac ion mix u e was
s i ed a . When he eac ion was comple e (TLC, 3:1 CHCl
3
/
MeOH) 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
and 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.
4.6. 4-(2,3,4,6-Te a-O-ace yl-b-
D
-glucopy anosyl)semica bazide
(2)
290
NH
2
NH
2
HOAc (97%, 26.6 mg, 0.28 mmol) was added o he
solu ion o 2,3,4,6- e a-O-ace yl-b-
D
-glucopy anosyl isocyana e
29
(1, 0.10 g, 0.27 mmol) and E
3
N (39
l
L, 0.28 mmol) in d y CH
2
Cl
2
(2 mL). The eac ion mix u e was s i ed a . When he eac ion
was comple e (TLC 5:1 E OAc/hexane) he sol en was e apo a ed
unde educed p essu e. The c ude p oduc was pu ified by column
ch oma og aphy (eluen : 5:1 E OAc/hexane) o gi e 81 mg (75%) o
2as a whi e amo phous p oduc . [
a
]
D
4(c0.50, DMSO); R
= 0.27
(5:1 E OAc/hexane);
1
H NMR (CDCl
3
, 360 MHz) d(ppm) 9.28, 6.82,
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])
A
O
HO
HO
HO
OH H
NH
N
X
N
H
CA
O
HO
HO
HO
OH H
NH
N
O
A
O
OH
HO
HO
OH
H
N
NN
OA
X=O X=S
10 38 — 34 18
4
24 20
Me
11 136 — — 25 12
O
2
N
12 4.5 31 25.7
18
(IC
50
)— 26 15
F
13 48 32 5.7
18
(IC
50
)— 27 14
Cl
14 30 33 23.2
18
(IC
50
)— 28 33
15 124 — 35 350
4
(IC
50
)29 315 (IC
50
)
16 5.5 — 36 5.2
4
30 27
O
HO
HO
HO
OH H
NH
N
O
X
O
O
HO
HO
HO
OH H
CN
H
NH
N
X
37 X = NH 21
4
39 X=O 29
19
38 X=CH
2
600
4
40 X = S 300
19
4M. Tó h e al. / Ca bohyd a e Resea ch xxx (2013) xxx–xxx
CAR 6468 No. o Pages 10, Model 5G
2 May 2013
Please ci e his a icle in p ess as: Tó h, M.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.04.025
6.53, 6.27 (4H, 4 s, NH), 5.32, 5.14, 5.07, 4.97 (4H, 4 p , J= 8.9,
300
9.6 Hz in each, H-1, H-2, H-3, H-4), 4.31–4.12 (2H, m, H-6a, H-
6b), 3.85–3.83 (1H, m, H-5), 2.08, 2.07, 2.04 (12H, 3 s, CH
3
).
13
C
NMR (CDCl
3
, 360 MHz) d(ppm) 170.0, 169.4, 169.3, 169.2 (CO),
157.2 (NHCONH), 78.5, 72.8, 71.7, 70.4, 67.9 (C-1 o C-5), 61.7
(C-6), 20.5, 20.3 (CH
3
). Anal. Calcd o C
15
H
23
N
3
O
10
(405.36): C,
44.44; H, 5.72; N, 10.37. Found: C, 44.58; H, 5.84; N, 10.26.
4.7. Benzaldehyde 4-(2,3,4,6- e a-O-ace yl-b-
D
-glucopy anosyl)
semica bazone (3)
F om isocyana e 1(0.10 g, 0.27 mmol) and benzaldehyde
hyd azone (0.064 g, 0.54 mmol) acco ding o Gene al p ocedu e I
310
(Sec ion 4.2). Pu ified by column ch oma og aphy (1:1 E OAc/
hexane) o yield 110 mg (83%) o 3as a whi e amo phous p oduc .
[
a
]
D
–61 (c0.99, CHCl
3
); R
: 0.58 (5:1 E OAc/hexane);
1
H NMR
(CDCl
3
, 360 MHz) d(ppm) 9.47 (1H, b s, NH), 7.77 (1H, s, CH@N),
7.70–7.66 (2H, m, A ), 7.43–7.40 (3H, m, A ), 7.16 (1H, d,
J
1,NH
= 9.5 Hz, NH), 5.39, 5.28, 5.15, 5.13 (4H, 4 p , J= 9.5, 9.7 Hz
in each, H-1, H-2, H-3, H-4), 4.38 (1H, dd, J
6a,6b
= 12.5 Hz, H-6a),
4.12 (1H, dd, H-6b), 3.92 (1H, ddd, J
5,6a
= 4.1 Hz, J
5,6b
= 1.8 Hz,
J
4,5
= 10.1 Hz, H-5), 2.08, 2.05, 2.04, 2.03 (12H, 4 s, CH
3
).
13
C NMR
(CDCl
3
, 360 MHz) d(ppm) 170.7, 170.5, 170.0, 169.5 (CO), 155.7
320
(NHCONH), 142.6 (CH@N), 133.6, 130.0, 128.7, 127.1 (A ), 79.3,
73.2, 72.8, 70.3, 68.2 (C-1 o C-5), 61.6 (C-6), 20.7, 20.6 (CH
3
). Anal.
Calcd o C
22
H
27
N
3
O
10
(493.46): C, 53.55; H, 5.51; N, 8.52. Found: C,
53.46; H, 5.61; N, 8.60.
4.8. 4-Me hylbenzaldehyde 4-(2,3,4,6- e a-O-ace yl-b-
D
-
glucopy anosyl)semica bazone (4)
F om isocyana e 1(0.40 g, 1.07 mmol) and 4-me hylbenzalde-
hyde hyd azone (0.29 g, 2.14 mmol) acco ding o Gene al p oce-
du e I (Sec ion 4.2). Pu ified by column ch oma og aphy (1:1
E OAc/hexane) o yield 390 mg (72%) o 4as a whi e amo phous
330
p oduc . [
a
]
D
–50 (c0.86, CHCl
3
); R
: 0.66 (5:1 E OAc/hexane);
1
H
NMR (CDCl
3
, 360 MHz) d(ppm) 9.21 (1H, b s, NH), 7.71 (1H, s,
CH@N), 7.56 (2H, d, J= 8.0 Hz, A ), 7.27–7.20 (2H, m, A ), 7.13
(1H, d, J
1,NH
= 9.5 Hz, NH), 5.37, 5.26, 5.14, 5.12 (4H, 4 p ,
J= 9.5 Hz in each, H-1, H-2, H-3, H-4), 4.36 (1H, dd, J
6a,6b
= 12.5 Hz,
H-6a), 4.11 (1H, dd, H-6b), 3.89 (1H, ddd, J
5,6a
= 4.1 Hz,
J
5,6b
= 1.9 Hz, J
4,5
= 10.1 Hz, H-5), 2.39, 2.08, 2.05, 2.04, 2.02 (16H,
5s,CH
3
).
13
C NMR (CDCl
3
, 360 MHz) d(ppm) 170.7, 170.5, 170.0,
169.5 (CO), 155.7 (NHCONH), 142.6 (CH@N), 140.3, 130.8, 129.5,
127.0 (A ), 79.4, 73.2, 72.9, 70.3, 68.2 (C-1 o C-5), 61.7 (C-6),
340
21.5, 20.7 (CH
3
). Anal. Calcd o C
23
H
29
N
3
O
10
(507.49): C, 54.43;
H, 5.76; N, 8.28. Found: C, 54.32; H, 5.64; N, 8.38.
4.9. 4-Ni obenzaldehyde 4-(2,3,4,6- e a-O-ace yl-b-
D
-
glucopy anosyl)semica bazone (5)
F om isocyana e 1(0.40 g, 1.07 mmol) and 4-ni obenzaldehyde
hyd azone (0.37 g, 2.14 mmol) acco ding o Gene al p ocedu e I
(Sec ion 4.2). Pu ified by column ch oma og aphy (1:1 E OAc/hex-
ane) o yield 380 mg (66%) o 5as an o ange amo phous p oduc .
[
a
]
D
71 (c0.35, CHCl
3
); R
: 0.18 (1:1 E OAc/hexane);
1
H NMR
(CDCl
3
, 400 MHz) d(ppm) 10.10 (1H, b s, NH), 8.29 (2H, d,
350
J= 8.8 Hz, A ), 7.87 (2H, d, J= 8.8 Hz, A ), 7.86 (1H, s, CH@N), 7.25
(1H, d, J
1,NH
= 9.2 Hz, NH), 5.41, 5.24, 5.14, 5.12 (4H, 4 p , J= 9.3,
9.8 Hz in each, H-1, H-2, H-3, H-4), 4.38 (1H, dd, J
6a,6b
= 12.5 Hz,
H-6a), 4.13 (1H, dd, H-6b), 3.91 (1H, J
5,6a
= 4.1 Hz, J
5,6b
= 2.1 Hz,
J
4,5
= 9.9 Hz, m, H-5), 2.09, 2.06, 2.04 (12H, 3 s, CH
3
).
13
C NMR
(CDCl
3
, 360 MHz) d(ppm) 170.7, 170.5, 169.9, 169.4 (CO), 155.7
(NHCONH), 148.2 (C–NO
2
), 139.8 (CH@N), 139.6, 127.5, 124.0
(A ), 79.3, 73.2, 72.5, 70.4, 68.1 (C-1 o C-5), 61.6 (C-6), 20.5
(CH
3
). Anal. Calcd o C
22
H
26
N
4
O
12
(538.46): C, 49.07; H, 4.87; N,
10.40. Found: C, 48.98; H, 4.80; N, 10.52.
360
4.10. 4-Fluo obenzaldehyde 4-(2,3,4,6- e a-O-ace yl-b-
D
-
glucopy anosyl)semica bazone (6)
F om isocyana e 1(0.25 g, 0.67 mmol) and 4-fluo obenzalde-
hyde hyd azone (0.19 g, 1.23 mmol) acco ding o Gene al p oce-
du e I (Sec ion 4.2). Pu ified by column ch oma og aphy (1:1
E OAc/hexane) o yield 260 mg (76%) o 6as a whi e amo phous
p oduc . [
a
]
D
–54 (c0.27, CHCl
3
); R
: 0.22 (1:1 E OAc/hexane);
1
H
NMR (CDCl
3
, 360 MHz) d(ppm) 10.09 (1H, b s, NH) 7.78 (1H, s,
CH@N), 7.68 (2H, dd, J= 5.6 Hz, J= 7.9 Hz, A ), 7.21–7.07 (3H, m,
A , NH), 5.41, 5.29, 5.15, 5.12 (4H, 4 p , J= 9.3, 9.9 Hz in each, H-
370
1, H-2, H-3, H-4), 4.38 (1H, dd, J
5,6a
= 3.7 Hz, J
6a,6b
= 12.5 Hz, H-
6a), 4.12 (1H, dd, J
5,6b
< 1 Hz, H-6b), 3.97–3.89 (1H, m, H-5), 2.05
(12H, s, CH
3
).
13
C NMR (CDCl
3
, 360 MHz) d(ppm) 170.6, 170.5,
169.9, 169.4 (CO), 163.7 (d, J= 250.0 Hz, A ), 155.8 (NHCONH),
141.4 (CH@N), 129.9 (A ), 128.9 (d, J= 7.3 Hz, A ), 115.8 (d,
J= 22.0 Hz, A ), 79.3, 73.1, 72.7, 70.3, 68.1 (C-1 o C-5), 61.6 (C-
6), 20.5 (CH
3
). Anal. Calcd o C
22
H
26
FN
3
O
10
(511.45): C, 51.66; H,
5.12; N, 8.22. Found: C, 51.54; H, 5.23; N, 8.33.
4.11. 3-Chlo obenzaldehyde 4-(2,3,4,6- e a-O-ace yl-b-
D
-
glucopy anosyl)semica bazone (7)
380
F om isocyana e 1(0.22 g, 0.59 mmol) and 3-chlo obenzalde-
hyde hyd azone (0.18 g, 1.18 mmol) acco ding o Gene al p oce-
du e I (Sec ion 4.2). Pu ified by column ch oma og aphy (1:1
E OAc/hexane) o yield 222 mg (71%) o 7as a whi e amo phous
p oduc . [
a
]
D
–65 (c1.06, CHCl
3
); R
: 0.61 (5:1 E OAc/hexane);
1
H
NMR (CDCl
3
, 360 MHz) d(ppm) 9.60 (1H, b s, NH), 7.64–7.46
(3H, m, A , CH@N), 7.31–7.27 (2H, m, A ), 7.12 (1H, d, J
1,NH
= 9.2 Hz,
NH), 5.31, 5.16, 5.07, 5.05 (4H, 4 p , J= 9.2, 9.7 Hz in each, H-1, H-2,
H-3, H-4), 4.29 (1H, dd, J
5,6a
= 3.9 Hz, J
6a,6b
= 12.4 Hz, H-6a), 4.06
(1H, dd, J
5,6b
= 1.0 Hz, H-6b), 3.87–3.80 (1H, m, H-5), 2.02, 1.97
390
(12H, 2 s, CH
3
).
13
C NMR (CDCl
3
, 360 MHz) d(ppm) 170.7, 170.5,
170.0, 169.5 (CO), 155.7 (NHCONH), 141.0 (CH@N), 135.4, 134.9,
130.0, 126.9, 125.3 (A ), 79.5, 73.3, 72.7, 70.4, 68.3 (C-1 o C-5),
61.7 (C-6), 20.7, 20.6 (CH
3
). Anal. Calcd o C
22
H
26
ClN
3
O
10
(527.91): C, 50.05; H, 4.96; N, 7.96. Found: C, 50.16; H, 4.84; N,
7.85.
4.12. 4-(1-Naph haldehyde) 4-(2,3,4,6- e a-O-ace yl-b-
D
-
glucopy anosyl)semica bazone (8)
F om isocyana e 1(0.40 g, 1.07 mmol) and 1-naph aldehyde
hyd azone (0.37 g, 2.14 mmol) acco ding o Gene al p ocedu e I
400
(Sec ion 4.2). Pu ified by column ch oma og aphy (1:1 E OAc/hex-
ane) o yield 520 mg (89%) o 8as a yellow amo phous p oduc . [
a
]
D
53 (c0.96, CHCl
3
); R
: 0.63 (5:1 E OAc/hexane);
1
H NMR (CDCl
3
,
360 MHz) d(ppm) 9.63 (1H, b s, NH), 8.52 (1H, d, J= 8.5 Hz, A ),
8.44 (1H, s, CH@N), 7.93–7.90 (3H, m, A ), 7.68 (1H, p , J= 8.1,
8.3 Hz, A ), 7.58–7.51 (2H, m, A ), 7.17 (1H, d, J
1,NH
= 9.6 Hz, NH),
5.42, 5.34, 5.17, 5.16 (4H, 4 p , J= 9.5, 9.7 Hz in each, H-1, H-2, H-
3, H-4), 4.37 (1H, dd, J
6a,6b
= 12.5 Hz, H-6a), 4.11 (1H, dd, H-6b),
3.94 (1H, ddd, J
5,6a
= 3.9 Hz, J
5,6b
= 2.0 Hz, J
4,5
= 10.2 Hz, H-5), 2.07,
2.06, 2.05 (12H, 3 s, CH
3
).
13
C NMR (CDCl
3
, 360 MHz) d(ppm)
410
170.7, 170.6, 170.0, 169.5 (CO), 155.7 (NHCONH), 142.1 (CH@N),
133.8, 130.5, 130.7, 128.9, 128.8, 127.3, 126.2, 125.3, 123.7 (A ),
79.4, 73.3, 72.9, 70.3, 68.2 (C-1 o C-5), 61.6 (C-6), 20.7, 20.6
(CH
3
). Anal. Calcd o C
26
H
29
N
3
O
10
(543.52): C, 57.45; H, 5.38; N,
7.73. Found: C, 57.56; H, 5.27; N, 7.85.
M. Tó h e al. / Ca bohyd a e Resea ch xxx (2013) xxx–xxx 5
CAR 6468 No. o Pages 10, Model 5G
2 May 2013
Please ci e his a icle in p ess as: Tó h, M.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.04.025
4.13. 4-(2-Naph haldehyde) 4-(2,3,4,6- e a-O-ace yl-b-
D
-gluco-
py anosyl)semica bazone (9)
F om isocyana e 1(0.27 g, 0.71 mmol) and 2-naph aldehyde
hyd azone (0.24 g, 1.42 mmol) acco ding o Gene al p ocedu e I
(Sec ion 4.2). Pu ified by column ch oma og aphy (1:1 E OAc/hex-
420
ane) o yield 310 mg (80%) o 9as a yellow amo phous p oduc . [
a
]
D
73 (c0.93, CHCl
3
); R
: 0.64 (5:1 E OAc/hexane);
1
H NMR (CDCl
3
,
360 MHz) d(ppm) 9.83 (1H, s, NH), 7.99–7.83 (6H, m, A , CH@N),
7.53–7.50 (2H, m, A ), 7.26 (1H, d, J
1,NH
= 9.8 Hz, NH), 5.42, 5.32,
5.17, (4H, 3 p , J= 9.3, 9.6 Hz in each, H-1, H-2, H-3, H-4), 4.39
(1H, dd, J
5,6a
= 3.8 Hz, J
6a,6b
= 12.5 Hz, H-6a), 4.14 (1H, dd,
J
5,6b
< 1.0 Hz, H-6b), 3.97–3.94 (1H, m, H-5), 2.06 (12H, s, CH
3
).
13
C NMR (CDCl
3
, 360 MHz) d(ppm) 170.7, 170.6, 170.0, 169.5
(CO), 155.7 (NHCONH), 142.1 (CH@N), 134.2, 133.1, 131.4, 128.7,
128.3, 127.9, 127.0, 126.6, 122.7 (A ), 79.5, 73.3, 72.9, 70.4, 68.3
430
(C-1 o C-5), 61.7 (C-6), 20.6 (CH
3
). Anal. Calcd o C
26
H
29
N
3
O
10
(543.52): C, 57.45; H, 5.38; N, 7.73. Found: C, 57.57; H, 5.49; N, 7.63.
4.14. Benzaldehyde 4-(b-
D
-glucopy anosyl)semica bazone (10)
F om 3(0.15 g, 0.30 mmol) acco ding o Gene al p ocedu e IV
(Sec ion 4.5). Pu ified by column ch oma og aphy (4:1 CHCl
3
/
MeOH) o yield 98 mg (99%) o 10 as a whi e amo phous p oduc .
[
a
]
D
+33 (c0.54, CH
3
OH); R
: 0.34 (4:1 CHCl
3
/MeOH);
1
H NMR
(D
2
O, 360 MHz) d(ppm) 7.82 (1H, s, CH@N), 7.72–7.68 (5H, m,
A ), 4.97 (1H, d, J
1,2
= 8.3 Hz, H-1), 3.90 (1H, dd, J
5,6b
= 1.5 Hz,
J
6a,6b
= 11.7 Hz, H-6b), 3.75 (1H, dd, J
5,6a
= 5.2 Hz, H-6a), 3.62–3.48
440
(4H, m, H-2, H-3, H-4, H-5).
13
C NMR (D
2
O+1 d op CH
3
OH,
360 MHz) d(ppm) 158.4 (NHCONH), 144.1 (CH@N), 135.7, 130.9,
129.7, 128.1 (A ), 82.4, 79.4, 78.8, 73.9, 71.4 (C-1 o C-5), 62.7 (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, 51.57; H, 5.99; N, 12.81.
4.15. 4-Me hylbenzaldehyde 4-(b-
D
-
glucopy anosyl)semica bazone (11)
F om 4(0.09 g, 0.18 mmol) acco ding o Gene al p ocedu e IV
(Sec ion 4.5). Pu ified by column ch oma og aphy (4:1 CHCl
3
/
MeOH) o yield 60 mg (98%) o 11 as a whi e amo phous p oduc .
450
[
a
]
D
+30 (c0.48, CH
3
OH); R
: 0.48 (3:1 CHCl
3
/MeOH);
1
H NMR
(CD
3
OD, 360 MHz) d(ppm) 7.85 (1H, s, CH@N), 7.59 (2H, d,
J= 8.5 Hz, A ), 7.20 (2H, d, J= 8.5 Hz, A ), 4.97 (1H, d, J
1,2
= 8.4 Hz,
H-1), 3.89 (1H, dd, J
5,6b
< 1.0 Hz, J
6a,6b
= 11.7 Hz, H-6b), 3.73 (1H,
dd, J
5,6a
= 3.8 Hz, H-6a), 3.48–3.35 (4H, m, H-2, H-3, H-4, H-5),
2.35 (3H, s, CH
3
).
13
C NMR (CD
3
OD, 360 MHz) d(ppm) 158.4
(NHCONH), 144.1 (CH@N), 141.3, 133.0, 130.4, 128.1 (A ), 82.4,
79.5, 78.9, 74.0, 71.4 (C-1 o C-5), 62.7 (C-6), 21.5 (CH
3
). Anal. Calcd
o C
15
H
21
N
3
O
6
(339.34): C, 53.09; H, 6.24; N, 12.38. Found: C,
52.98; H, 6.35; N, 12.26.
460
4.16. 4-Ni obenzaldehyde 4-(b-
D
-glucopy anosyl)
semica bazone (12)
F om 5(0.15 g, 0.28 mmol) acco ding o Gene al p ocedu e IV
(Sec ion 4.5). Pu ified by column ch oma og aphy (1:1 CHCl
3
/
MeOH) o yield 107 mg (93%) o 12 as an o ange amo phous p od-
uc . [
a
]
D
+30 (c0.29, DMSO); R
: 0.70 (2:3 CHCl
3
/MeOH);
1
H NMR
(CD
3
OD, 400 MHz) d(ppm) 8.28 (2H, d, J= 8.7 Hz, A ), 8.00 (2H,
d, J= 9.0 Hz, A ), 7.98 (1H, s, CH@N), 4.94 (1H, d, J
1,2
= 8.5 Hz, H-
1), 3.84 (1H, dd, J
5,6b
= 1.0 Hz, J
6a,6b
= 12.3 Hz, H-6b), 3.70 (1H, dd,
J
5,6a
= 4.5 Hz, J
6a,6b
= 11.9 Hz, H-6a), 3.51–3.43 (2H, m, H-2 o H-3
470
o H-4, H-5), 3.40–3.36 (2H, m, H-2 and/o H-3, and/o H-4).
13
C
NMR (DMSO-d
6
,360 MHz) d(ppm) 155.0 (NHCONH), 147.3 (C–
NO
2
), 138.0 (CH@N), 141.0, 127.8, 129.8 (A ), 80.9, 78.4, 77.4,
71.9, 69.9 (C-1 o C-5), 61.0 (C-6). Anal. Calcd o C
14
H
18
N
4
O
8
(370.31): C, 45.41; H, 4.90; N, 15.13. Found: C, 45.54; H, 4.79; N,
15.24.
4.17. 4-Fluo obenzaldehyde 4-(b-
D
-glucopy anosyl)
semica bazone (13)
F om 6(0.12 g, 0.24 mmol) acco ding o Gene al p ocedu e IV
(Sec ion 4.5). Pu ified by column ch oma og aphy (4:1 CHCl
3
/
480
MeOH) o yield 80 mg (98%) o 13 as a b own amo phous p oduc .
[
a
]
D
+42 (c0.28, DMSO); R
: 0.20 (4:1 CHCl
3
/MeOH);
1
HNMR
(CD
3
OD, 400 MHz) d(ppm) 7.89 (1H, s, CH@N), 7.78 (2H, dd,
J= 5.6 Hz, J= 8.5 Hz, A ), 7.15 (2H, p , J= 8.7 Hz, A ), 4.95 (1H, d,
J
1,2
= 8.4 Hz, H-1), 3.86 (1H, dd, J
5,6b
< 1.0 Hz, J
6a,6b
= 11.7 Hz, H-
6b), 3.70 (1H, dd, J
5,6a
= 4.0 Hz, H-6a), 3.48–3.29 (4H, m, H-2, H-3,
H-4, H-5).
13
C NMR (DMSO-d
6
,360 MHz) d(ppm) 162.8 (d,
J= 246.4 Hz, A ), 155.4 (NHCOHN), 138.0 (CH@N), 131.2, 129.2 (d,
J= 6.4 Hz, A ), 115.8 (d, J= 21.6 Hz, A ), 80.9, 78.4, 77.5, 72.1, 70.1
(C-1 o C-5), 61.1 (C-6). Anal. Calcd o C
14
H
18
FN
3
O
6
(343.31): C,
490
48.98; H, 5.28; N, 12.24. Found: C, 49.10; H, 5.41; N, 12.13.
4.18. 3-Chlo obenzaldehyde 4-(b-
D
-glucopy anosyl)
semica bazone (14)
F om 7(0.15 g, 0.28 mmol) acco ding o Gene al p ocedu e IV
(Sec ion 4.5). Pu ified by column ch oma og aphy (4:1 CHCl
3
/
MeOH) o yield 96 mg (94%) o 14 as a yellow amo phous p oduc .
[
a
]
D
+30 (c0.54, CH
3
OH); R
: 0.40 (4:1 CHCl
3
/MeOH);
1
HNMR
(CD
3
OD, 360 MHz) d(ppm) 7.83–7.32 (5H, m, A , CH@N), 4.97
(1H, d, J
1,2
= 8.3 Hz, H-1), 3.85 (1H, dd, J
5,6b
< 1.0 Hz, J
6a,6b
= 11.8 Hz,
H-6b), 3.71–3.67 (1H, m, H-6a), 3.52–3.39 (4H, m, H-2, H-3, H-4, H-
500
5).
13
C NMR (CD
3
OD, 360 MHz) d(ppm) 158.2 (NHCONH), 142.4
(CH@N), 137.8, 135.8, 131.2, 130.6, 127.3, 126.9 (A ), 82.4, 79.4,
78.8, 73.8, 71.3 (C-1 o C-5), 62.6 (C-6). Anal. Calcd o
C
14
H
18
ClN
3
O
6
(359.76): C, 46.74; H, 5.04; N, 11.68. Found: C,
46.62; H, 5.13; N, 11.80.
4.19. 1-Naph haldehyde 4-(b-
D
-glucopy anosyl)semica bazone
(15)
F om 8(0.17 g, 0.18 mmol) acco ding o Gene al p ocedu e IV
(Sec ion 4.5). Pu ified by column ch oma og aphy (4:1 CHCl
3
/
MeOH) o yield 93 mg (80%) o 15 as a b own amo phous p oduc .
510
[
a
]
D
+32 (c0.48, CH
3
OH); R
: 0.41 (4:1 CHCl
3
/MeOH);
1
HNMR
(CD
3
OD, 360 MHz) d(ppm) 8.66 (1H, s, CH@N), 8.41–7.48 (7H, m,
A ), 4.98 (1H, d, J
1,2
= 8.7 Hz, H-1), 3.87 (1H, dd, J
5,6b
= 1.5 Hz,
J
6a,6b
= 12.0 Hz, H-6b), 3.71 (1H, dd, J
5,6a
= 4.6 Hz, H-6a), 3.52–3.38
(4H, m, H-2, H-3, H-4, H-5).
13
C NMR (CD
3
OD, 360 MHz) d(ppm)
158.3 (NHCONH), 142.3 (CH@N), 135.3–124.1 (A ), 82.4, 79.5,
79.0, 74.1, 71.4 (C-1 o C-5), 62.7 (C-6). Anal. Calcd o
C
18
H
21
N
3
O
6
(375.38): C, 57.59; H, 5.64; N, 11.19. Found: C, 57.70;
H, 5.52; N, 11.29.
4.20. 2-Naph haldehyde 4-(b-
D
-glucopy anosyl)semica bazone
520
(16)
F om 9(0.14 g, 0.26 mmol) acco ding o Gene al p ocedu e IV
(Sec ion 4.5). Pu ified by column ch oma og aphy (4:1 CHCl
3
/
MeOH) o yield 96 mg (98%) o 16 as a whi e amo phous p oduc .
[
a
]
D
+39 (c0.47, CH
3
OH); R
: 0.43 (4:1 CHCl
3
/MeOH);
1
HNMR
(CD
3
OD, 360 MHz) d(ppm) 8.08–7.46 (8H, m, A , CH@N), 4.97
(1H, d, J
1,2
= 8.6 Hz, H-1), 3.87 (1H, dd, J
5,6b
< 1.0 Hz, J
6a,6b
= 12.0 Hz,
H-6b), 3.71 (1H, dd, J
5,6a
= 4.3 Hz, H-6a), 3.51–3.38 (4H, m, H-2, H-
3, H-4, H-5).
13
C NMR (CD
3
OD, 360 MHz) d(ppm) 157.9 (NHCONH),
143.9 (CH@N), 135.6–123.9 (A ), 82.4, 79.5, 78.9, 74.0, 71.4 (C-1 o
530
C-5), 62.7 (C-6). Anal. Calcd o C
18
H
21
N
3
O
6
(375.38): C, 57.59; H,
5.64; N, 11.19. Found: C, 57.46; H, 5.75; N, 11.08.
6M. Tó h e al. / Ca bohyd a e Resea ch xxx (2013) xxx–xxx
CAR 6468 No. o Pages 10, Model 5G
2 May 2013
Please ci e his a icle in p ess as: Tó h, M.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.04.025
4.21. 5-Phenyl-2-(2,3,4,6- e a-O-ace yl-b-
D
-
glucopy anosylamino)-1,3,4-oxadiazole (17)
(a) F om 3(0.11 g, 0.23 mmol) acco ding o Gene al p ocedu e II
(Sec ion 4.3). Pu ified by column ch oma og aphy (1:1 E OAc/hex-
ane) o yield 50 mg (45%) o 17 as a whi e amo phous p oduc .
(b) F om 3(0.25 g, 0.51 mmol) acco ding o Gene al p ocedu e
III (Sec ion 4.4). Pu ified by column ch oma og aphy (1:1 E OAc/
hexane) o yield 212 mg (85%) o 17 as a whi e amo phous p oduc .
540
[
a
]
D
–20 (c0.28, CHCl
3
); R
: 0.23 (1:1 E OAc/hexane);1H NMR
(CDCl
3
, 360 MHz) d(ppm) 7.96–7.83 (2H, m, A ), 7.54–7.42 (3H,
m, A ), 6.22 (1H, b s, NH), 5.39 (1H, p , J= 9.4 Hz, H-2 o H-3 o
H-4), 5.21 (1H, d, J
1,2
= 9.0 Hz, H-1), 5.14, 5.11 (2H, 2 p , J= 9.4,
9.9 Hz in each, H-2 and/o H-3, and/o H-4), 4.33 (1H, dd,
J
5,6a
= 4.5 Hz, J
6a,6b
= 12.5 Hz, H-6a), 4.11 (1H, dd, J
5,6b
= 1.2 Hz, H-
6b), 3.98–3.88 (1H, m, H-5), 2.09, 2.06, 2.05 (12H, 3 s, CH
3
).
13
C
NMR (CDCl
3
, 360 MHz) d(ppm) 170.8, 170.5, 169.8, 169.5 (CO),
161.4, 160.0 (C-oxadiazole), 131.0, 128.9, 125.0, 123.8 (A ), 82.7,
73.4, 72.6, 70.6, 68.1 (C-1 o C-5), 61.6 (C-6), 20.6, 20.5 (CH
3
). Anal.
550
Calcd o C
22
H
25
N
3
O
10
(491.45): C, 53.77; H, 5.13; N, 8.55. Found: C,
53.89; H, 5.24; N, 8.64.
4.22. 5-(4-Me hylphenyl)-2-(2,3,4,6- e a-O-ace yl-b-
D
-
glucopy anosylamino)-1,3,4-oxadiazole (18)
F om 4(0.29 g, 0.57 mmol) acco ding o Gene al p ocedu e III
(Sec ion 4.4). Pu ified by column ch oma og aphy (1:1 E OAc/hex-
ane) o yield 190 mg (66%) o 18 as a whi e amo phous p oduc . [
a
]
D
–14 (c0.27, CHCl
3
); R
: 0.31 (1:1 E OAc/hexane);1H NMR (CDCl
3
,
360 MHz) d(ppm) 7.78 (2H, d, J= 8.1 Hz, A ), 7.24 (2H, d,
J= 8.0 Hz, A ), 6.40 (1H, b s, NH), 5.38 (1H, p , J= 9.4 Hz, H-2 o
560
H-3 o H-4), 5.21 (1H, d, J
1,2
= 9.3 Hz, H-1), 5.14, 5.12 (2H, 2 p ,
J= 9.3, 9.7 Hz in each, H-2 and/o H-3, and/o H-4), 4.31 (1H, dd,
J
6a,6b
= 12.5 Hz, H-6a), 4.09 (1H, dd, H-6b), 3.92 (1H, ddd,
J
5,6a
= 4.2 Hz, J
5,6b
= 1.8 Hz, J
4,5
= 9.8 Hz, H-5), 2.39, 2.07, 2.04, 2.03
(16H, 4 s, CH
3
).
13
C NMR (CDCl
3
, 360 MHz) d(ppm) 170.6, 170.5,
169.8, 169.4 (CO), 161.2, 160.0 (C-oxadiazole), 141.3, 129.5, 125.9,
121.0 (A ), 82.6, 73.2, 72.7, 70.5, 68.0 (C-1 o C-5), 61.6 (C-6), 21.4
(CH
3
), 20.6, 20.5 (CH
3
). Anal. Calcd o C
23
H
27
N
3
O
10
(505.47): C,
54.65; H, 5.38; N, 8.31. Found: C, 54.78; H, 5.49; N, 8.42.
4.23. 5-(4-Ni ophenyl)-2-(2,3,4,6- e a-O-ace yl-b-
D
-
570
glucopy anosylamino)-1,3,4-oxadiazole (19)
(a) F om 5(0.19 g, 0.35 mmol) acco ding o Gene al p oce-
du e II (Sec ion 4.3). Pu ified by column ch oma og aphy (1:1
E OAc/hexane) o yield 80 mg (43%) o 19 as a whi e amo phous
p oduc .
(b) F om 5(0.25 g, 0.46 mmol) acco ding o Gene al p ocedu e
III (Sec ion 4.4). Pu ified by column ch oma og aphy (1:1 E OAc/
hexane) o yield 130 mg (53%) o 19 as a whi e amo phous p oduc .
[
a
]
D
–21 (c0.23, CHCl
3
); R
: 0.27 (1:1 E OAc/hexane);
1
H NMR
(CDCl
3
, 360 MHz) d(ppm) 8.34 (2H, d, J= 8.6 Hz, A ), 8.10 (2H, d,
580
J= 8.6 Hz, A ), 6.45 (1H, b s, NH), 5.41 (1H, p , J= 9.4 Hz, H-2 o
H-3 o H-4), 5.24 (1H, d, J
1,2
= 9.0 Hz, H-1), 5.16, 5.12 (2H, 2 p ,
J= 9.7, 10.0 Hz in each, H-2 and/o H-3, and/o H-4), 4.34 (1H,
dd, J
5,6a
= 4.3 Hz, J
6a,6b
= 12.5 Hz, H-6a), 4.14 (1H, dd, J
5,6b
< 1.0 Hz,
H-6b), 4.00–3.91 (1H, m, H-5), 2.10, 2.07 (12H, 2 s, CH
3
).
13
C
NMR (CDCl
3
, 360 MHz) d(ppm) 170.9, 170.5, 169.8, 169.5 (CO),
162.1, 158.2 (C-oxadiazole), 148.9 (C–NO
2
), 129.3, 126.7, 124.3
(A ), 82.6, 73.4, 72.5, 70.6, 68.0 (C-1 o C-5), 61.5 (C-6), 20.6, 20.5
(CH
3
). Anal. Calcd o C
22
H
24
N
4
O
12
(536.45): C, 49.26; H, 4.51; N,
10.44. Found: C, 49.37; H, 4.42; N, 10.33.
590
4.24. 5-(4-Fluo ophenyl)-2-(2,3,4,6- e a-O-ace yl-b-
D
-
glucopy anosylamino)-1,3,4-oxadiazole (20)
(a) F om 6(0.20 g, 0.39 mmol) acco ding o Gene al p ocedu e II
(4.3). Pu ified by column ch oma og aphy (1:1 E OAc/hexane) o
yield 48 mg (24%) o 20 as a whi e amo phous p oduc .
(b) F om 6(0.1 g, 0.19 mmol) acco ding o Gene al p ocedu e III
(4.4). Pu ified by column ch oma og aphy (1:1 E OAc/hexane) o
yield 85 mg (88%) o 20 as a whi e amo phous p oduc . [
a
]
D
=–4
(c0.30, CHCl
3
); R
: 0.33 (1:1 E OAc/hexane); 1H NMR (CDCl
3
,
360 MHz) d(ppm) 7.91 (2H, dd, J= 5.4 Hz, J= 8.2 Hz, A ), 7.16
600
(2H, p , J= 8.4 Hz A ), 6.27 (1H, b s, NH), 5.39 (1H, p , J= 9.4 Hz,
H-2 o H-3 o H-4), 5.21 (1H, d, J
1,2
= 9.2 Hz, H-1), 5.14, 5.10 (2H,
2 p , J= 9.3, 9.8 Hz in each, H-2 and/o H-3, and/o H-4), 4.34
(1H, dd, J
5,6a
= 4.3 Hz, J
6a,6b
= 12.3 Hz,H-6a), 4.12 (1H, dd,
J
5,6b
< 1.0 Hz, H-6b), 3.98–3.88 (1H, m, H-5), 2.09, 2.06, 2.05 (12H,
3s, CH
3
).
13
C NMR (CDCl
3
, 360 MHz) d(ppm) 170.8, 170.5, 169.8,
169.5 (CO), 164.1 (d, J= 252.0 Hz, A ), 161.4, 159.1 (C-oxadiazole),
128.2 (d, J= 8.0 Hz, A ), 120.1, 116.2 (d, J= 22.3 Hz, A ), 82.6, 73.3,
72.6, 70.5, 68.0 (C-1 o C-5), 61.5 (C-6), 20.6, 20.5 (CH
3
). Anal. Calcd
o C
22
H
24
FN
3
O
10
(509.44): C, 51.87; H, 4.75; N, 8.25. Found: C,
610
51.76; H, 4.87; N, 8.38.
4.25. 5-(3-Chlo ophenyl)-2-(2,3,4,6- e a-O-ace yl-b-
D
-
glucopy anosylamino)-1,3,4-oxadiazole (21)
(a) F om 7(0.13 g, 0.24 mmol) acco ding o Gene al p ocedu e II
(Sec ion 4.3). Pu ified by column ch oma og aphy (1:1 E OAc/hex-
ane) o yield 65 mg (52%) o 21 as a whi e amo phous p oduc .
(b) F om 7(0.17 g, 0.33 mmol) acco ding o Gene al p ocedu e
III (Sec ion 4.4). Pu ified by column ch oma og aphy (1:1 E OAc/
hexane) o yield 110 mg (64%) o 21 as a whi e amo phous p oduc .
[
a
]
D
–13 (c0.32, CHCl
3
); R
: 0.29 (1:1 E OAc/hexane);
1
HNMR
620
(CDCl
3
, 360 MHz) d(ppm) 7.89 (1H, b s, A ), 7.80 (1H, d,
J= 7.3 Hz, A ), 7.51–7.36 (2H, m, A ), 6.22 (1H, b s, NH), 5.39
(1H, p , J= 9.4 Hz, H-2 o H-3 o H-4), 5.21 (1H, d, J
1,2
= 9.2 Hz, H-
1), 5.14, 5.10 (2H, 2 p , J= 9.4, 9.8 Hz in each, H-2 and/o H-3,
and/o H-4), 4.33 (1H, dd, J
5,6a
= 4.4 Hz, J
6a,6b
= 12.4 Hz, H-6a),
4.13 (1H, dd, J
5,6b
< 1.0 Hz, H-6b), 3.99–3.90 (1H, m, H-5), 2.09,
2.06, 2.05 (12H, 3 s, CH
3
).
13
C NMR (CDCl
3
, 360 MHz) d(ppm)
170.9, 170.5, 169.8, 169.5 (CO), 161.5, 158.8 (C-oxadiazole),
135.0, 131.0, 130.3, 125.9, 125.4, 124.1 (A ), 82.6, 73.4, 72.6, 70.6,
68.1 (C-1 o C-5), 61.6 (C-6), 20.6, 20.5 (CH
3
). Anal. Calcd o
630
C
22
H
24
ClN
3
O
10
(525.89): C, 50.25; H, 4.60; N, 7.99. Found: C,
50.38; H, 4.71; N, 7.89.
4.26. 5-(1-Naph hyl)-2-(2,3,4,6- e a-O-ace yl-b-
D
-
glucopy anosylamino)-1,3,4-oxadiazole (22)
F om 8(0.34 g, 0.63 mmol) acco ding o Gene al p ocedu e III
(Sec ion 4.4). Pu ified by column ch oma og aphy (1:1 E OAc/hex-
ane) o yield 305 mg (89%) o 22 as a whi e amo phous p oduc .
[
a
]
D
–10 (c0.29, CHCl
3
); R
: 0.32 (1:1 E OAc/hexane);
1
HNMR
(CDCl
3
, 360 MHz) d(ppm) 9.01 (1H, d, J= 8.4 Hz, A ), 7.86–7.69
(3H, m, A ), 7.52–7.26 (3H, m, A ), 6.94 (1H, b s, NH), 5.32 (1H,
640
p , J= 9.2 Hz, H-2 o H-3 o H-4), 5.19 (1H, d, J
1,2
= 9.0 Hz, H-1),
5.09, 5.06 (2H, 2 p , J= 9.5, 9.8 Hz in each, H-2 and/o H-3, and/o
H-4), 4.22 (1H, dd, J
5,6a
= 4.0 Hz, J
6a,6b
= 12.7 Hz, H-6a), 4.02 (1H,
dd, J
5,6b
< 1.0 Hz, H-6b), 3.90–3.80 (1H, m, H-5), 1.98, 1.93, 1.90
(12H, 3 s, CH
3
).
13
C NMR (CDCl
3
, 360 MHz) d(ppm) 170.3, 170.2,
169.7, 169.3 (CO), 161.2, 159.5 (C-oxadiazole), 133.4–120.1 (A ),
82.4, 73.1, 72.7, 70.4, 67.9 (C-1 o C-5), 61.5 (C-6), 20.3 (CH
3
). Anal.
Calcd o C
26
H
27
N
3
O
10
(541.51): C, 57.67; H, 5.03; N, 7.76. Found: C,
57.55; H, 5.12; N, 7.63.
M. Tó h e al. / Ca bohyd a e Resea ch xxx (2013) xxx–xxx 7
CAR 6468 No. o Pages 10, Model 5G
2 May 2013
Please ci e his a icle in p ess as: Tó h, M.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.04.025