Synthesis of substituted 2-(β-d-glucopyranosyl)-benzimidazoles and their evaluation as inhibitors of glycogen phosphorylase

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Please check his box i you ha e no co ec ions o make o he PDF ile G aphical abs ac pp xxx–xxxSyn hesis o subs i u ed 2-(b- DD -glucopy anosyl)-benzimidazoles and hei e alua ion as inhibi o s o glycogen phospho ylase É a Boko , Enik} o Szilágyi, Tibo Docsa, Pa ´l Ge gely, La ´szlo ´Somsa ´k * CAR 6384 No. o Pages 1, Model 5G 5 Feb ua y 2013 Highligh s "Syn hesis o C-(b- DD -glucopy anosyl) o mimida es and - hio o mimida es. "New syn hesis o C-(b- DD -glucopy anosyl)-benzimidazoles. "Low mic omola inhibi o s o glycogen phospho ylase. X=O,S O OR 1 R 1 O R 1 O OR 1 N H N O OBz BzO BzO COOH OBz O OBz BzO BzO CXNH 2 OBz O OBz BzO BzO C OBz NH XE H 2 N H 2 N R 2 E 3 OBF 4 om X=O R 2 R 1 =Bz H NO 2 O OH HO HO OH N H N bes inhibi o K i =2.1μM( abbi mu s R 2 =5-F,5-Cl,5-B ,5- N 4-Me, 5-Me, 5,6-di 1 Syn hesis o subs i u ed 2-(b-D-glucopy anosyl)-benzimidazoles and hei e alua ion as inhibi o s o glycogen phospho ylase É a Boko a ,Enik} oSzilágyi 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 3 No embe 2012 Recei ed in e ised o m 15 Janua y 2013 Accep ed 16 Janua y 2013 A ailable online xxxx Keywo ds: C-Glycopy anosyl- o mimida e C-Glycopy anosyl- hio o mimida e 2-C-Glycopy anosyl benzimidazole Glycogen phospho ylase Inhibi o abs ac Mic owa e assis ed condensa ion o O-pe benzoyla ed C-(b-D-glucopy anosyl) o mic acid wi h 1,2-diaminobenzenes in he p esence o iphenylphosphi e ga e he co esponding O-p o ec ed 2-(b-D-glucopy anosyl)-benzimidazoles in mode a e yields. O-Pe benzoyla ed C-(b-D-glucopy anosyl) o mamide and - hio o mamide we e ans o med in o he co esponding e hyl C-(b-D-glucopy anosyl) o mimida e and - hio o mimida e, espec i ely, by E 3 OBF 4 . T ea men o he o mimida e wi h 1,2-diaminobenzenes a o ded O-p o ec ed 2-(b-D-glucopy anosyl)-benzimidazoles-in good o excellen yields. Simila eac ion o he hio o mimida e ga e hese compounds in lowe yields. The O-benzoyl p o ec ing g oups we e emo ed by he Zemplén p o ocol. These es compounds we e assayed agains abbi muscle glycogen phospho ylase (GP) b, he p o o ype o li e GP, he a e limi ing enzyme o glycogen deg ada ion. The bes inhibi o s we e 2-(b-D-glucopy anosyl)-4-me hyl-benzimidazole (K i = 2.8 l M) and 2-(b-D-glucopy anosyl)-naph ho[2,3-d]imidazole (K i = 2.1 l M) exhibi ing a 3–4 imes s onge binding han he unsubs i u ed pa en compound. Ó2013 Published by Else ie L d. 1. In oduc ion Inhibi ion o glycogen phospho ylase (GP) has been conside ed as an e ec i e he apeu ic app oach in comba ing ype 2 diabe es ( o he biochemical and pha macological backg ound o a ge ing li e GP as a alida ed concep o lowe ing blood glucose le els, please su ey ecen e iew a icles 1–6 ). Fu he mo e, he pha ma- ceu ical u ili y o GP inhibi o s in he in e en ion o o he diseased s a es associa ed wi h GP ac i i y (e.g., ca dio ascula dis- 50 o de s, 7–9 ischaemic lesions, 10–13 and umo ous g ow h 11,14–16 ) has also been unde in es iga ion. A la ge a ay o compounds we e shown o ha e an inhibi o y e ec agains his enzyme 17–19 including glucose de i a i es 20,21 which p ima ily bind o he ca aly ic si e o he enzyme. In he cou se o sea ching o po en glucose based inhibi o s, se e al C-b- D -glucopy anosyl he e ocycles (Cha 1), such as e azole 22 1A, 1,3,4-oxadiazoles 22–24 1B, 1,2,4-oxadiazoles 23,25,26 1C,D, benzo- hiazole 22 1E and benzimidazole 22 1F ha e been syn hesized and some o hem p o ed o be e ficien agains abbi muscle glycogen 60 phospho ylase b(RMGPb, he p o o ype o GP enzymes o enzy- ma ic es s 2 ). In his class, 2-(b- D -glucopy anosyl)-benzimidazole (1F) was he fi s compound o ha e a K i alue in he low mic omo- la ange. 22,27 As e idenced by X- ay c ys allog aphy, he s ong binding in he ca aly ic cen e is he esul o di ec and wa e media ed H-bonds be ween he p o ein and he he e oa oma ic ing, and an de Waals in e ac ions o he la ge a oma ic pa in he so-called b-channel o he enzyme. 27 The highe a fini y o 1F in compa ison wi h i s hio coun e pa 1E can be a ibu ed o he di ec H-bond o he imidazole NH wi h he His377 main chain 70 ca bonyl g oup o he enzyme which is ce ainly absen o benzo- hiazole 1E. 27 Analogous NH-(His377)CO in e ac ions we e iden i- fied in o he GP enzyme-inhibi o complexes (e.g., in cases o spi o( hio)hydan oin 28,29 and N-acyl-b- D -glucopy anosylamine ype inhibi o s 30 ) indica ing he p ominen impo ance o his spe- cial H-b idge, as well. Addi ionally, X- ay c ys allog aphic in es i- ga ion o 1F in complex wi h RMGPb e ealed ha besides he ac i e si e he compound also occupied he new allos e ic si e, and a new binding cle called he benzimidazole si e was also disco e ed. 27 80 To ge an insigh in o he s uc u e–ac i i y ela ionship o his ype o inhibi o he aim o ou p esen wo k has been o syn he- size a se ies o subs i u ed 2-(b- D -glucopy anosyl)-benzimidazoles and o e alua e hei e ec on RMGPb. Fo he o ma ion o 2-C-glycosyl-benzimidazoles se e al syn- he ic me hods a e known om he li e a u e mos o which ha e been desc ibed o u anose based de i a i es. Fu anosyl benzimi- dazoles we e p epa ed (a) by acid ca alysed condensa ion o C-gly- co u anosyl o mic acids and o-phenylenediamine (OPD), 31,32 (b) by coupling o C-glyco u anosyl o mic acids o hei chlo ides wi h 90 1,2-diaminobenzenes, ollowed by acid o POCl 3 media ed ing closu e o he esul ing amide ype in e media es, 33,34 (c) in he 0008-6215/$ - see on ma e Ó2013 Published by Else ie L d. h p://dx.doi.o g/10.1016/j.ca es.2013.01.011 ⇑ Co esponding au ho . Tel.: +36 52512900x22348; ax: +36 52512744. E-mail add ess: [email p o ec ed] (L. Somsák). Q1 Q2 Q3 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 6384 No. o Pages 9, Model 5G 5 Feb ua y 2013 Please ci e his a icle in p ess as: Boko ,É.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.01.011 eac ion o C-glyco u anosyl o maldoximoyl chlo ides (ac ually he ni ile oxide ob ained by base induced dehyd ochlo ina ion) wi h OPD, 35 (d) by in amolecula ing closu e o sui ably p o ec ed 2-(pen i ol-1 0 -yl)-benzimidazoles ob ained om 2-li hi- a ed benzimidazoles and suga lac one o acyclic pen ose de i a- i es, 36–38 and (e) by acid induced cyclodehyd a ion o unp o ec ed 2-( e i ol-1 0 -yl)-benzimidazoles. 39 Fo he syn hesis o 2-C-glycopy anosyl-benzimidazoles h ee 100 p ocedu es we e epo ed: (a) acid ca alysed condensa ion o unp o ec ed C-glycopy anosylme hanals (gene a ed in si u om he app op ia e dime hyl ace als) wi h OPD, ollowed by spon ane- ous oxida ion o he in e media e benzimidazolines, 40 (b) eac ion o O-pe benzoyla ed e hyl (C-b- D -glucopy anosyl) hio o mimida e hyd ochlo ide (p epa ed om he co esponding glycosyl cyanide) wi h OPD, 22 and (c) ea men o O-pe ace yla ed C-glycopy anosyl o maldoximoyl chlo ides wi h OPD. 41 In addi ion, a 2-benzimidaz- olyl moie y was also a ached o C-1 o D -galac al in he eac ion o me hyl C-(2-deoxy- D -lyxo-hex-1-enopy anosyl) o mimida e and 110 he dihyd ochlo ide sal o OPD. 42 In his pape , we disclose u he syn he ic possibili ies o con- s uc benzimidazole a he anome ic cen e o he py anose uni . 2. Resul s and discussion 2.1. Syn heses C-(b- D -Glucopy anosyl) o mic acid 5, p epa ed om cyanide 2 ia amide 3 43 by he li e a u e p o ocol 44 (Table 1), was eac ed wi h 1,2-diaminobenzenes (a, ,h) in he p esence o iphenylphosphi e in py idine unde mic owa e i adia ion (condi ions we e adap ed om a published p ocedu e applied o non-suga based com- 120 pounds 45 ). Al hough he con e sions we e comple e a 140 °Cin 20 min, he desi ed p oduc s could be isola ed only in mode a e yields (8a: 45%, 8 : 53%, 8h: 43%). Ne e heless, i has o be no ed, ha ea lie a emp s o ans o m 5in o benzimidazole 8a by he classical acid ca alysed p ocedu e wi h con en ional hea ing b ough abou no eac ion e en a ele a ed empe a u es. 22 The e o e, applica ion o he mo e eac i e iminoes e s 6and 7 was en isaged o he cons uc ion o benzimidazoles. P e iously, hyd ochlo ide sal o O-pe benzoyla ed e hyl C-(b- D -glucopy anosyl) hio o mimida e ob ained om glucopy - 130 anosyl cyanide 2by an acid ca alysed addi ion o E SH o he ni ile g oup was used o his pu pose. 22 Since hese condi ions we e a he unpleasan and subsequen ans o ma ion o he sal ga e benzimidazole 8a in a 34% yield only, we se ou o p oduce and use he ee o m o his ype o p ecu so . F ee hioimida e 7 was ob ained om hioamide 4by E 3 OBF 4 . The p epa a ion o 4 46 was also modified o a oid he use o H 2 S; hus, cyanide 2 was eac ed wi h P 4 S 10 in efluxing E OH as desc ibed o he syn- hesis o alipha ic and a oma ic hioamides. 47 Simila ly o 7, imi- da e 6was ob ained om amide 3by E 3 OBF 4 . 140 Nex , he p epa a ion o benzimidazole 8a om ei he 6o 7 was compa ed in p elimina y expe imen s. Reac ion o 6o 7wi h OPD ga e 8a in 89% and 62% yields, espec i ely. The e o e, also aking in o accoun he ins abili y o 7(decomposi ion was obse ed on s o age a a e a ew days), u he eac ions o ge he desi ed benzimidazoles we e pe o med wi h 6. Simila ly o 8a, high yields we e also achie ed in eac ions o 6 wi h me hyl-subs i u ed 1,2-diaminobenzenes –h and 2,3-diami- nonaph halene i, espec i ely (Table 1). A compa ison o he yields o 8a, ,h ob ained om acid 5unde condi ions and om imi- 150 da e 6unde condi ions i, espec i ely, showed he ing closu e o 6wi h 1,2-diaminobenzenes o be supe io o ha o 5. On ea men o imida e 6wi h 1,2-diaminobenzenes con ain- ing elec on wi hd awing subs i uen s (b–e) benzoic acid elimina- ion was also obse ed, and beside he 2-glucopy anosyl benzimidazoles 8b–e 4-subs i u ed-2-(3 0 ,4 0 ,6 0 - i-O-benzoyl-2 0 - deoxy- D -a abino-hex-1-enopy anosyl)-benzimidazoles (1-C-benz- imidazolyl glucals, 9b–e) we e also isola ed. Fu he mo e, o al consump ion o he s a ing ma e ial 6in eac ion wi h 4-ni o-1,2-diaminobenzene (e) equi ed highe empe a u e, hus 160 cycliza ion was pe o med in boiling e hanol o yield benzimid- azole 8e oge he wi h 9e. This endency o elimina ion migh be a ibu ed o an inc eased acidi y o he C-1–H p o on in he 2-glucosyl-benzimidazoles p obably due o he p esence o he elec on wi hd awing subs i uen in he a oma ic ing sys em. Fo enzyma ic s udies emo al o he benzoyl p o ec ing g oups o 8b–i was e ec ed by he Zemplén me hod o gi e es com- pounds 10b–i in good yields. The s uc u e o he new compounds was de e mined by 1 H and 13 C NMR spec oscopy. The p esence o a b- D -configu ed glucopy - 170 anosyl moie y in he 4 C 1 con o ma ion was confi med by he ici- nal p o on–p o on coupling cons an s o compounds 6,7,8b–i and 10b–i, espec i ely. In he 1 H NMR spec a o 1-C-he a yl glu- cals 9b–e he obse ed small couplings be ween he icinal ing p o ons (3–6 Hz) sugges ed a con o ma ional change o he py a- nose ing ( om chai o hal chai ). 48 In he 13 C NMR spec a o 9b–e cha ac e is ic esonances appea ed o C-1 0 (143–145 ppm) and C-2 0 (99–101 ppm) p o iding e idence o he unsa u a ed na u e o he py anoid ing. 48 Among 13 C NMR signals o he benz- imidazole uni s only hose o C-2 appea ed as sha p peaks a 146– 180 158 ppm, he o he s usually ga e b oad signals. These obse a ions a e in acco d wi h he li e a u e expe iences, 35,41 and may be ex- plained by he apid p o on exchange be ween N-1 and N-3 o he benzimidazole moie y. 35,41,49 Addi ionally, in he 13 CNMR spec a duplica ion o ce ain ca bon peaks was also obse ed (p i- ma ily o he benzimidazole ca bon signals o example, in 8b,9c, 9e and 10b), which may e e o he coexis ence o he au ome ic pai s. Fo he fluo o-benzimidazole 8b his phenomenon seemed qui e clea as 14 esonances we e obse ed in he 13 C NMR spec- um which could be en a i ely assigned on he basis o he cha - 190 ac e is ic C–F couplings 50 as illus a ed in Figu e 1. O OH HO HO C OH 1 He R He R K i [μM] A NN NHN - No inhibi ion 22 B O NN R CH 3 145 27 212 22 2-naph hyl 10 % a 625 μM 23 C N ON R 2-naph hyl 38 25 D N NO R 2-naph hyl 2.4 23 E S N -76 27 229 22 F N H N -8.6 27 11 22 Cha 1. Inhibi o y po ency (K i )o C-glucosyl he e ocyclic de i a i es agains abbi muscle glycogen phospho ylase b(RMGPb). 2É a Boko e al. /Ca bohyd a e Resea ch xxx (2013) xxx–xxx CAR 6384 No. o Pages 9, Model 5G 5 Feb ua y 2013 Please ci e his a icle in p ess as: Boko ,É.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.01.011 2.2. Enzyme kine ic s udies Enzyme kine ic measu emen s wi h abbi muscle glycogen phospho ylase bwe e pe o med as desc ibed p e iously, 51,52 and he ob ained inhibi o cons an s a e gi en in Table 2. The new compounds showed inhibi o y e ec s in he mic omola ange and some o hem p o ed equi alen o somewha be e inhibi o s han 10a (=1F in Cha 1). Among benzimidazoles wi h a halogen subs i uen in he 5-po- si ion fluo o de i a i e 10b exhibi ed modes ac i i y, while he 200 chlo o (10c) as well as b omo (10d) compounds we e as good inhibi o s as 10a. Subs i u ion o he benzimidazole ing in he same posi ion by a ni o g oup (10e) b ough abou a significan dec ease o he inhibi ion. Among he alkyl subs i u ed de i a i es he 5-me hyl- (10 ) and 4-me hyl-benzimidazoles (10g) displayed simila and sligh ly be e (3- old inc ease) po ency han 10a, espec i ely. Howe e , in oduc ion o wo me hyl g oups in o he 5- and 6-posi ions o he he e ocycle (10h) esul ed in a ema kable weakening o he inhibi ion. Finally, he naph ho[2,3- d]imidazole 10i showed 4 imes s onge binding han 10a and 210 p o ed o be he mos e ec i e inhibi o o his se ies. To elucida e he na u e o he binding modes o he new compounds o GP enzyme X- ay c ys allog aphic s udies a e in p og ess and will be epo ed elsewhe e. Table 1 Syn hesis o 2-(b-D-glucopy anosyl)-benzimidazoles 3X=O(94%) 4X=S(66%) + 6X=O(96%) 7X=S(90%) 8a-i R 1 =Bz 9a-i a-i 5(76 %) iii i i O OR 1 R 1 O R 1 O OR 1 N H N R 2 R 3 R 4 O OBz BzO BzO COOH OBz O OBz BzO BzO CXNH 2 OBz O OBz BzO BzO C OBz NH XE O BzO BzO OBz N H N R 2 R 3 R 4 10a-i R 1 =H ii 2 O OBz BzO BzO CN OBz H 2 N H 2 N R 2 R 3 R 4 i o 3 ii o 4 i) HB -AcOH, ;43 ii) P4S10, abs. E OH, e lux; iii) NO2, abs. CH2Cl2, ;44 i ) E 3O·BF4, abs. CH2Cl2, A , ; ) P(OPh)3 (1.2 equi .), a o o h (1.0 equi ), abs. py idine, μW, 140 ºC, 20 min; i) a-i (2 equi .), abs. CH2Cl2, e lux; ii) ~1 M NaOMe in abs. MeOH, . Reagen R 2 R 3 R 4 Condi ions and yields (%) 8910 aHH H 45 — ii — c,d i 89 ( om 6)— i 62 a ( om 7) bHF H i 46 35 ii 54 cHCl H i 76 24 ii 89 dHB H i 54 29 ii 58 eHNO 2 H i b 46 20 ii 53 HMeH 53 — ii 87 i 83 — gMe H H i 79 — ii 73 hHMeMe 43 — ii 78 i 81 — iH i 79 — ii 77 a Repo ed yield o 8a by he ea lie me hod om pe benzoyla ed b- D -glucopy anosyl cyanide ia he hioimida e hyd ochlo ide sal : 34%. 22 b Modified eac ion condi ions we e necessa y o comple e consump ion o he s a ing ma e ial 6: abs. E OH, eflux. c Debenzoyla ion o 8a by he Zemplén p o ocol was ca ied ou ea lie . Repo ed yield o 10a: 84%. 22 d Repo ed yield o 10a by a di e en me hod: 54%. 40 N N H F Bz 4 -β-D-Glc p (H) 1(3) 3(1) 3a(7a) 4(7) 5(6) 6(5) 7(4) 7a(3a) 2(2) 151.1 (s) 150.3 (s) 158.9 (d, 1 J C-F =237Hz) 158.2 (d, 1 J C-F =237Hz) 104.5 (d, 2 J C-F =24Hz) 98.0 (d, 2 J C-F =27Hz) 142.8 (d, 3 J C-F ~9Hz) 134.2 (d, 3 J C-F ~9Hz) 139.0 (s) 130.8 (s) 110.9 (d, 2 J C-F =23Hz) 109.7 (d, 2 J C-F =23Hz) 120.0 (d, 3 J C-F ~7Hz) 112.3 (d, 3 J C-F ~6Hz) in e changable assignmen s Figu e 1. Ten a i e assignmen o 13 C NMR signals o au ome s o compound 8b. É a Boko e al. /Ca bohyd a e Resea ch xxx (2013) xxx–xxx 3 CAR 6384 No. o Pages 9, Model 5G 5 Feb ua y 2013 Please ci e his a icle in p ess as: Boko ,É.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.01.011 3. Conclusion New syn he ic pa hways we e elabo a ed o he o ma ion o subs i u ed 2-(b- D -glucopy anosyl)-benzimidazoles by he e ocyc- liza ions o 2,6-anhyd o-aldonic acid de i a i es as p ecu so s. Condensa ion o O-pe benzoyla ed C-(b- D -glucopy anosyl) o mic acid wi h 1,2-diaminobenzenes in he p esence o P(OPh) 3 unde 220 mic owa e i adia ion ga e he desi ed benzimidazoles in mode - a e yields. Ring closu e o O-pe benzoyla ed e hyl C-(b- D -glucopy - anosyl) o mimida e, p epa ed om he co esponding o mamide by E 3 OBF 4 , wi h he 1,2-diaminobenzenes a o ded a mo e e ficien p ocedu e. Wi h elec on deple ed a oma ic diamines undesi able b-elimina ion o benzoic acid om he 1,2-posi ions o he glucopy anosyl moie y also ook place o yield 1-C-benzim- idazolyl glucals besides he a ge compounds. Enzyme kine ic s udies wi h he new 2-(b- D -glucopy anosyl)-benzimidazoles showed he compounds o inhibi RMGPbin he low mic omola 230 ange. Subs i u ion o he benzimidazole moie y by a me hyl g oup in he 4-posi ion o u he annela ion o a benzene ing p o ided mo e e ficien inhibi o s han he unsubs i u ed pa en compound. 4. Expe imen al 4.1. Gene al me hods Mel ing poin s we e measu ed 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–El- me 241 pola ime e a . NMR spec a we e eco ded wi h B uke 360 (360/90 MHz o 1 H/ 13 C) spec ome e . Chemical shi s a e e - e enced o Me 4 Si ( 1 H), o o he esidual sol en signals ( 13 C). 240 Mic owa e assis ed p ocedu es we e pe o med in a CEM-Disco e Focused Mic owa e Syn hesis Sys em (2450 MHz) wi h a buil -in in a ed empe a u e senso and a CEM-Explo e compu e con- olled obo ic sample . TLC was pe o med on DC-Alu olle Kieselgel 60 F 254 (Me ck), and he pla es we e isualized unde 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 used. Dichlo ome hane was dis iled om P 4 O 10 and s o ed o e 4 Å molecula sie es. 2,3,4,6-Te a-O-benzoyl-b- D -glucopy anosyl cyanide, 43 C-(2,3,4, 250 6- e a-O-benzoyl-b- D -glucopy anosyl) o mamide 43 and C-(2,3,4, 6- e a-O-benzoyl-b- D -glucopy anosyl) o mic acid 44 we e syn he- sized acco ding o published p ocedu es. 4.2. C-(2,3,4,6-Te a-O-benzoyl-b- D -glucopy anosyl) hio o mamide (4) A solu ion o phospho us pen asulfide (2.23 g, 10 mmol) in anhyd ous E OH (20 mL) was s i ed a o 1.5 h, hen 2,3,4,6- e - a-O-benzoyl-b- D -glucopy anosyl cyanide 43 (2, 3.02 g, 5 mmol) was added and he mix u e was hea ed a eflux empe a u e o 2h.A e cooling he he e ogenous mix u e o , he p ecipi a e 260 was fil e ed o and washed wi h E OH o yield 2.1 g (66%) o 4 as a pale yellowish solid. Mp: 199–201 °C (Li . 46 mp: 198– 200 °C). 1 H and 13 C NMR da a co espond o he epo ed spec a. 46 4.3. E hyl C-(2,3,4,6- e a-O-benzoyl-b- D -glucopy anosyl) o mimida e (6) C-(2,3,4,6-Te a-O-benzoyl-b- D -glucopy anosyl) o mamide 43 (3, 1.0 g, 1.60 mmol) and E 3 OBF 4 (0.91 g, 4.80 mmol) we e dissol ed in anhyd ous CH 2 Cl 2 (15 mL), he mix u e was s i ed a unde A and moni o ed by TLC (1:1 E OAc–hexane). A e comple ion o he eac ion (2 days), he mix u e was dilu ed wi h CH 2 Cl 2 270 (30 mL), ex ac ed wi h sa d aq NaHCO 3 solu ion (30 mL) and hen wi h wa e (30 mL). The o ganic phase was d ied o e MgSO 4 ,fil- e ed and he sol en was e apo a ed. The c ude pale yellow amo - phous p oduc (1.0 g, 96%) was used wi hou u he pu ifica ion. R : 0.55 (1:1 E OAc–hexane); 1 H NMR (CDCl 3 )d(ppm): 8.06–7.25 (21H, m, a oma ics, NH), 5.99, 5.72, 5.55 (3 1H, 3 pseudo , J= 10.6, 9.2 Hz in each, H-2, H-3, H-4), 4.70 (1H, dd, J= 11.9, 2.6 Hz, H-6a), 4.52 (1H, dd, J= 11.9, 5.3 Hz, H-6b), 4.25 (1H, ddd, J= 9.2, 5.3, 2.6 Hz, H-5), 4.21 (1H, d, J= 9.2 Hz, H-1), 4.12–3.91 (2H, m, CH 2 ), 0.84 (3H, , 6.6 Hz, CH 3 ); 13 C NMR (CDCl 3 )d(ppm): 280 167.5, 166.1, 165.7, 165.1 (2) (CO, CNH), 133.5–128.2 (a oma ics), 75.8, 74.6, 73.6, 70.7, 69.1 (C-1–C-5), 62.8, 62.1 (C-6, CH 2 ), 13.4 (CH 3 ). 4.4. E hyl C-(2,3,4,6- e a-O-benzoyl-b- D -glucopy anosyl) hio o mimida e (7) C-(2,3,4,6-Te a-O-benzoyl-b- D -glucopy anosyl) hio o mamide (4, 0.2 g, 0.31 mmol) and E 3 OBF 4 (0.18 g, 0.94 mmol) we e dis- sol ed in anhyd ous CH 2 Cl 2 (4 mL), he mix u e was s i ed a unde A and moni o ed by TLC (1:1 E OAc–hexane). A e comple- ion o he eac ion (2 days), he mix u e was dilu ed wi h CH 2 Cl 2 290 (10 mL), ex ac ed wi h sa d aq NaHCO 3 solu ion (15 mL), hen wi h wa e (15 mL). The o ganic phase was d ied o e MgSO 4 ,fil- e ed and he sol en was e apo a ed. The c ude pale yellow amo - phous p oduc (0.19 g, 90%) was used wi hou u he pu ifica ion. R : 0.62 (1:1 E OAc–hexane); 1 H NMR (CDCl 3 )d(ppm): 8.06–7.24 (21H, m, a oma ics, NH), 5.95, 5.74, 5.70 (3 1H, 3 pseudo , J= 9.8, 9.1 Hz in each, H-2, H-3, H-4), 4.68 (1H, dd, J= 11.9, 2.8 Hz, H-6a), 4.51 (1H, dd, J= 11.9, 4.9 Hz, H-6b), 4.39 (1H, d, J= 9.1 Hz, H-1), 4.22 (1H, ddd, J= 9.1, 4.9, 2.8 Hz, H-5), 2.88–2.71 (2H, m, CH 2 ), 1.17 (3H, pseudo , 7.7, 7.0 Hz, CH 3 ); 13 C NMR (CDCl 3 ) d(ppm): 172.7 (CNH), 166.1, 165.7, 165.1, 164.9 (CO), 133.4–128.2 (a oma ics), 80.6, 76.1, 73.9, 70.9, 69.1 (C-1–C-5), 62.8 (C-6), 23.1 (CH 2 ), 13.1 (CH 3 ). 4.5. Gene al p ocedu e I o he syn hesis o subs i u ed 2-(2 0 ,3 0 ,4 0 ,6 0 - e a-O-benzoyl-b- D -glucopy anosyl)- benzimidazoles om O-pe benzoyla ed C-(b- D -glucopy anosyl) o mic acid (5) C-(2,3,4,6-Te a-O-benzoyl-b- D -glucopy anosyl) o mic acid 44 (5, 0.1 g, 0.16 mmol), a 1,2-diaminobenzene (ao o h, 0.16 mmol, 1equi ) and iphenyl phosphi e (50 l l, 0.19 mmol, 1.2 equi ) 310 we e dissol ed in anhyd ous py idine (2 mL). The closed ial was i adia ed by mic owa es o 20 min a 140 °C (maximum p es- Table 2 Kine ic da a on he inhibi ion o abbi muscle glycogen phospho ylase bby he new compounds 10b–i O OH HO HO OH N H N R 2 R 3 R 4 10 R 2 R 3 R 4 K i ( l M) aH H H 8.6 27 11 22 bHF H55 cH Cl H 9.7 dH B H 7.5 eHNO 2 H 179 HMeH12 gMe H H 2.8 hH Me Me 152 iH2.1 Q4 4É a Boko e al. /Ca bohyd a e Resea ch xxx (2013) xxx–xxx CAR 6384 No. o Pages 9, Model 5G 5 Feb ua y 2013 Please ci e his a icle in p ess as: Boko ,É.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.01.011 su e: 20 ba , powe 220 W). The comple ion o he eac ion was moni o ed by TLC (1:1 E OAc–hexane). The eac ion mix u e was concen a ed unde educed p essu e, 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). 4.6. Gene al p ocedu e II o he syn hesis o subs i u ed 2-(2 0 ,3 0 ,4 0 ,6 0 - e a-O-benzoyl-b- D -glucopy anosyl)- 320 benzimidazoles om O-pe benzoyla ed e hyl (C-b- D -glycopy anosyl) o mimida e (6) E hyl C-(2,3,4,6- e a-O-benzoyl-b- D -glycopy anosyl) o mimi- da e (6, 0.1 g, 0.15 mmol) and a 1,2-diaminobenzene (a–i, 0.30 mmol, 2 equi ) we e dissol ed in anhyd ous CH 2 Cl 2 (3 mL). The mix u e was efluxed and moni o ed by TLC (2:3 E OAc–hex- ane). A e comple ion o he eac ion he sol en was e apo a ed, and he esidue was pu ified by column ch oma og aphy. 4.7. Gene al p ocedu e III o he Zemplén-deacyla ion A benzoyla ed compound was dissol ed in d y MeOH (5 mL/ 330 100 mg, a ew d ops o CHCl 3 we e added in case o incomple e dis- solu ion) and a ca aly ic amoun o a NaOMe solu ion (1M in MeOH) was added. The mix u e was kep a and moni o ed by TLC (7:3 CHCl 3 –MeOH). When he s a ing ma e ial was consumed he mix u e was neu alised wi h a ca ion exchange esin Ambe - lys 15 (H + o m), hen he esin was fil e ed o and he sol en e- mo ed. The esidue was pu ified by column ch oma og aphy. 4.8. 2-(2 0 ,3 0 ,4 0 ,6 0 -Te a-O-benzoyl-b- D -glucopy anosyl)- benzimidazole (8a) A: F om acid 5(0.1 g, 0.16 mmol) and o-phenylenediamine (a, 340 0.02 g, 0.16 mmol) acco ding o Gene al p ocedu e I. Yield: 0.05 g (45%). B: F om imida e 6(0.10 g, 0.15 mmol) and o-phenylenediamine (a, 0.04 g, 0.30 mmol) acco ding o Gene al P ocedu e II. Reac ion ime: 3 h. Pu ified by column ch oma og aphy (2:3 E OAc–hexane) o yield 0.09 g (89%) o whi e solid. Mp: 122–124 °C (Li . 22 mp: 120–123 °C). 1 H and 13 C NMR da a co espond o he epo ed spec a. 22 4.9. 5(6)-Fluo o-2-(2 0 ,3 0 ,4 0 ,6 0 - e a-O-benzoyl-b- D - glucopy anosyl)-benzimidazole (8b) and 5(6)-fluo o-2-(3 0 ,4 0 ,6 0 - 350 i-O-benzoyl-2 0 -deoxy- D -a abino-hex-1-enopy anosyl)- benzimidazole (9b) F om imida e 6(0.10 g, 0.15 mmol) and 1,2-diamino-4-fluo o- benzene (b, 0.04 g, 0.30 mmol) acco ding o Gene al P ocedu e II. Reac ion ime: 5 h. Pu ified by column ch oma og aphy (1:2, hen 2:3 E OAc–hexane) o gi e 9b as he fi s han 8b as he second ac ion. Compound 8b: Yield: 0.051 g (46%) pale yellow solid; R : 0.31 (2:3 E OAc–hexane); mp: 124–126 °C; [ a ] D =39 (c0.5, DMSO); 1 H NMR (CDCl 3 )d(ppm): 11.58 (1H, b s, benzimidazole NH), 360 7.96–6.76 (23H, m, a oma ics), 6.30–6.21, 6.02 (3 1H, 3 pseudo , J= 9.2, 9.2 Hz in each, H-2 0 , H-3 0 , H-4 0 ), 5.36 (1H, d, J= 9.2 Hz, H-1 0 ), 4.69 (1H, dd, J= 12.3, <1 Hz, H-6 0 a), 4.57 (1H, dd, J= 12.3, 5.3 Hz, H-6 0 b), 4.52 (1H, ddd, J= 9.2, 5.3, <1 Hz, H-5 0 ); 13 C NMR (DMSO-d 6 )d(ppm): 165.4, 165.1, 164.8, 164.3 (CO), 158.9 (d, 1 J C-F = 237 Hz, benzimidazole), 158.2 (d, 1 J C-F = 237 Hz, benz- imidazole), 151.1, 150.3 (benzimidazole), 142.8 (d, 3 J C-F = 9 Hz, benzimidazole), 139.0 (b s, benzimidazole), 134.2 (d, 3 J C-F = 9 Hz, benzimidazole), 133.7–133.3 (a oma ics), 130.9 (b s, benzimid- azole), 129.3–128.5 (a oma ics), 120.0 (d, 3 J C-F = 7 Hz, benzimid- 370 azole), 112.3 (d, 3 J C-F = 6 Hz, benzimidazole), 110.9 (d, 2 J C-F = 23 Hz, benzimidazole), 109.7 ( 2 J C-F = 23 Hz, benzimidazole), 104.5 (d, 2 J C-F = 24 Hz, benzimidazole), 98.0 (d, 2 J C-F = 27 Hz, benzimid- azole), 79.1, 74.8, 74.2, 73.4, 71.5, 69.0 (C-1 0 –C-5 0 ), 62.8 (C-6 0 ). Anal. Calcd o C 41 H 31 FN 2 O 9 (714.69): C, 68.90; H, 4.37; N, 3.92. Found: C, 68.84; H, 4.29; N, 4.03. Compound 9b: Yield: 0.032 g (35%), pale yellow sy up; R : 0.58 (2:3 E OAc–hexane); [ a ] D =7(c0.5, CHCl 3 ); 1 H NMR (CDCl 3 )d (ppm): 10.41 (1H, b s, benzimidazole NH), 8.02–6.94 (18H, m, a o- ma ics), 6.43 (1H, b s, H-2 0 ), 5.95–5.92 (2H, m, H-3 0 , H-4 0 ), 4.88 380 (1H, ddd, J= 4.0, <1 Hz, H-5 0 ), 4.79 (2H, s, H-6 0 a, H-6 0 b); 13 CNMR (CDCl 3 )d(ppm): 166.2, 165.6, 165.0 (CO), 159.8 (d, 1 J C-F = 239 Hz, benzimidazole C-5), 146.8 (benzimidazole C-2), 145.0 (C-1 0 ), 143.1, 139.8 (2 b s, benzimidazole C-3a, C-7a), 133.5–128.4 (a oma ics), 120.2 (b s, benzimidazole C-4 o C-7), 111.9 (d, 2 J C-F = 25 Hz, benzimidazole C-6), 104.4 (b s, benzimidazole C-4 o C-7), 99.4 (C-2 0 ), 75.2, 67.9, 67.3 (C-3 0 –C-5 0 ), 61.7 (C-6 0 ). Analysis: C 34 H 25 FN 2 O 7 (592.57), ESI-MS (posi i e mode) m/z: 615.157 [M+Na] + , 1207.327 [2 M+Na] + . 4.10. 5(6)-Chlo o-2-(2 0 ,3 0 ,4 0 ,6 0 - e a-O-benzoyl-b- D - 390 glucopy anosyl)-benzimidazole (8c) and 5(6)-chlo o-2-(3 0 ,4 0 ,6 0 - i-O-benzoyl-2 0 -deoxy- D -a abino-hex-1-enopy anosyl)- benzimidazole (9c) F om imida e 6(0.10 g, 0.15 mmol) and 1,2-diamino-4-chlo o- benzene (c, 0.04 g, 0.30 mmol) acco ding o Gene al P ocedu e II. Reac ion ime: 5 h. Pu ified by column ch oma og aphy (2:3 E OAc–hexane) o gi e 9c as he fi s and 8c as he second ac ion. Compound 8c: Yield: 0.085 g (76%), pale yellow solid; R : 0.29 (2:3 E OAc–hexane); mp: 123–125 °C; [ a ] D =61 (c0.5, CHCl 3 ); 1 H NMR (CDCl 3 )d(ppm): 11.33 (1H, b s, benzimidazole NH), 400 7.94–6.95 (23H, m, a oma ics), 6.21, 6.14, 6.00 (3 1H, 3 pseudo , J= 9.4, 9.4 Hz in each, H-2 0 , H-3 0 , H-4 0 ), 5.31 (1H, d, J= 9.4 Hz, H-1 0 ), 4.72 (1H, dd, J= 12.3, <1 Hz, H-6 0 a), 4.60 (1H, dd, J= 12.3, 4.4 Hz, H-6 0 b), 4.50 (1H, ddd, J= 9.4, 4.4, <1 Hz, H-5 0 ); 13 CNMR (CDCl 3 )d(ppm): 166.5, 166.0, 165.1 (2) (CO), 149.8 (benzimidazole C-2), 142.0, 137.8 (2 b s, benzimidazole C-3a, C-7a), 133.3–127.8 (a oma ics), 126.9, 123.3, 119.3 (b s), 112.4 (b s) (benzimidazole C-4–C-7), 77.0, 75.5, 74.1, 71.7, 69.5 (C-1 0 –C-5 0 ), 63.4 (C-6 0 ). Anal. Calcd o C 41 H 31 ClN 2 O 9 (731.15): C, 67.35; H, 4.27; N, 3.84. Found: C, 67.47; H, 4.13; N, 3.90. 410 Compound 9c: Yield: 0.022 g (24%), pale yellow sy up; R : 0.59 (2:3 E OAc–hexane); [ a ] D =5(c0.5, CHCl 3 ); 1 H NMR (CDCl 3 )d (ppm): 11.40 (1H, b s, benzimidazole NH), 8.00–7.07 (18H, m, a o- ma ics), 6.45 (1H, d, J= 4.0 Hz, H-2 0 ), 5.98 (1H, pseudo , J= 6.6, 5.9 Hz, H-3 0 o H-4 0 ), 5.94 (1H, pseudo , J= 5.9, 4.0 Hz, H-3 0 o H-4 0 ), 4.81 (1H, ddd, J= 6.6, 4.6, 4.0 Hz, H-5 0 ), 4.71–4.70 (2H, m, H-6 0 a, H-6 0 b); 13 C NMR (CDCl 3 )d(ppm): 166.1, 165.6, 164.9 (CO), 146.8 (b s, benzimidazole C-2), 145.0 (C-1 0 ), 144.1, 141.8 (2 b s, benzimidazole C-3a, C-7a), 133.6–133.2 (a oma ics), 132.0 (b s, benzimidazole C-5), 129.8–128.3 (a oma ics), 123.9, 120.3, 119.1, 420 112.2, 111.3 (b s o each, benzimidazole C-4, C-6, C-7), 99.8 (C-2 0 ), 75.2, 68.1, 67.3 (C-3 0 –C-5 0 ), 61.7 (C-6 0 ). Analysis: C 34 H 25 ClN 2 O 7 (609.02), ESI-MS (posi i e mode) m/z:631.123 [M+Na] + , 1239.257 [2M+Na] + . 4.11. 5(6)-B omo-2-(2 0 ,3 0 ,4 0 ,6 0 - e a-O-benzoyl-b- D - glucopy anosyl)-benzimidazole (8d) and 5(6)-b omo-2-(3 0 ,4 0 ,6 0 - i-O-benzoyl-2 0 -deoxy- D -a abino-hex-1-enopy anosyl)- benzimidazole (9d) F om imida e 6(0.10 g, 0.15 mmol) and 1,2-diamino-4-b omo- benzene (d, 0.06 g, 0.30 mmol) acco ding o Gene al P ocedu e II. 430 Reac ion ime: 3 h. Pu ified by column ch oma og aphy (2:3 E OAc–hexane) o gi e 9d as he fi s and 8d as he second ac ion. Q5 É a Boko e al. /Ca bohyd a e Resea ch xxx (2013) xxx–xxx 5 CAR 6384 No. o Pages 9, Model 5G 5 Feb ua y 2013 Please ci e his a icle in p ess as: Boko ,É.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.01.011 Compound 8d: Yield: 0.064 g (54%) pale yellow solid; R : 0.31 (2:3 E OAc–hexane); mp: 126–128 °C; [ a ] D =62 (c0.5, CHCl 3 ); 1 H NMR (CDCl 3 )d(ppm): 8.02–6.76 (23H, m, a oma ics), 6.42, 6.29, 6.14 (3 1H, 3 pseudo , J= 9.6, 9.6 Hz in each, H-2 0 , H-3 0 , H-4 0 ), 5.39 (1H, d, J= 9.6 Hz, H-1 0 ), 4.72 (1H, dd, J= 12.3, 2.2 Hz, H-6 0 a), 4.63 (1H, dd, J= 12.3, 4.9 Hz, H-6 0 b), 4.56 (1H, ddd, J= 9.6, 4.9, 2.2 Hz, H-5 0 ); 13 C NMR (CDCl 3 )d(ppm): 166.4, 166.0, 165.0 (2) (CO), 149.6 (benzimidazole C-2), 140.9, 138.1 (2 b s, benzimid- 440 azole C-3a, C-7a), 133.3–127.7 (a oma ics), 125.9, 119.0 (b s), 117.3 (b s), 115.9 (benzimidazole C-4, C-5, C-6, C-7), 77.0, 75.5, 74.1, 71.7, 69.4 (C-1 0 –C-5 0 ), 63.4 (C-6 0 ). Analysis: C 41 H 31 B N 2 O 9 (775.60), ESI-MS (posi i e mode) m/z:799.112 [M+Na] + , 1573.242 [2M+Na] + . Compound 9d: Yield: 0.029 g (29%) pale yellow sy up; R : 0.61 (2:3 E OAc–hexane); [ a ] D =4(c0.25, CHCl 3 ); 1 H NMR (CDCl 3 )d (ppm): 10.06 (1H, b s, imidazole NH), 8.04–7.33 (18H, m, a oma - ics), 6.46 (1H, b s, H-2 0 ), 5.97–5.93 (2H, m, H-3 0 , H-4 0 ), 4.91 (1H, ddd, J= 4.0, <1 Hz, H-5 0 ), 4.81 (2H, s, H-6 0 a, H-6 0 b). Analysis: 450 C 34 H 25 B N 2 O 7 (653.48), ESI-MS (posi i e mode) m/z:677.074 [M+Na] + , 1329.197 [2M+Na] + . 4.12. 5(6)-Ni o-2-(2 0 ,3 0 ,4 0 ,6 0 - e a-O-benzoyl-b- D - glucopy anosyl)-benzimidazole (8e) and 5(6)-ni o-2-(3 0 ,4 0 ,6 0 - i-O-benzoyl-2 0 -deoxy- D -a abino-hex-1-enopy anosyl)- benzimidazole (9e) F om imida e 6(0.10 g, 0.15 mmol) and 1,2-diamino-4-ni o- benzene (e, 0.05 g, 0.30 mmol) acco ding o Gene al p ocedu e II. in anhyd ous e hanol. Reac ion ime: 1 day. Pu ified by column ch oma og aphy (1:2 E OAc–hexane) o gi e 9e as he fi s and 460 8e as he second ac ion. Compound 8e: Yield: 0.052 g (46%) yellow solid; R : 0.25 (2:3 E OAc–hexane); mp: 134–136 °C; [ a ] D =65 (c0.5, CHCl 3 ); 1 H NMR (CDCl 3 )d(ppm): 12.87 (1H, b s, benzimidazole NH), 8.45– 6.96 (23H, m, a oma ics), 6.20, 6.12, 6.01 (3 1H, 3 pseudo , J= 9.4, 9.4 Hz in each, H-2 0 , H-3 0 , H-4 0 ), 5.33 (1H, d, J= 9.4 Hz, H-1 0 ), 4.77 (1H, dd, J= 11.8, <1 Hz, H-6 0 a), 4.63 (1H, dd, J= 11.8, <1 Hz, H-6 0 b), 4.51 (1H, ddd, J=J= 9.4, <1 Hz, H-5 0 ); 13 C NMR (CDCl 3 )d(ppm): 166.3, 165.8, 165.1, 164.8 (CO), 152.7 (benzimid- azole C-2), 143.3 (benzimidazole C-5), 141.6, 138.0 (2 b s, benz- 470 imidazole C-3a, C-7a), 133.5–127.6 (a oma ics), 118.9, 116.1, 111.6 (b s o each, benzimidazole C-4, C-6, C-7), 77.1, 75.4, 74.0, 71.7, 69.0 (C-1 0 –C-5 0 ), 63.2 (C-6 0 ). Anal. Calcd o C 41 H 31 N 3 O 11 (741.70): C, 66.39; H, 4.21; N, 5.67. Found: C, 66.26; H, 4.13; N, 5.79. Compound 9e:Yield: 0.019 g (20%) yellow sy up; R : 0.39 (2:3 E OAc–hexane); [ a ] D =+1 (c0.5, CHCl 3 ); 1 H NMR (DMSO-d 6 )d (ppm): 13.51 (1H, b s, benzimidazole NH), 8.13–7.48 (18H, m, a o- ma ics), 6.40 (1H, b s, H-2 0 ), 6.02 (1H, pseudo , J= 5.3, 4.0 Hz, H-3 0 o H-4 0 ), 5.93 (1H, pseudo , J= 6.6, 5.9 Hz, H-3 0 o H-4 0 ), 5.23 (1H, 480 ddd, J= 5.9, 5.3, 3.3 Hz, H-5 0 ), 4.89 (1H, dd, J= 12.6, 5.3 Hz, H-6 0 a), 4.78 (1H, dd, J= 12.6, 3.3 Hz, H-6 0 b); 13 C NMR (DMSO-d 6 )d (ppm): 165.3, 165.0, 164.5 (CO), 150.5 (b s), 149.7 (b s), 147.5 (b s), 145.0, 143.0, 142.3 (b s), 138.7 (b s) (benzimidazole, C-1 0 ), 133.8–128.7 (a oma ics), 119.2, 118.5, 117.8, 115.2, 112.3, 108.4 (b s o each, benzimidazole), 100.5 (C-2 0 ), 74.6, 67.9, 67.0 (C-3 0 –C-5 0 ), 61.5 (C-6 0 ). Analysis: C 34 H 25 N 3 O 9 (619.58), ESI-MS (po- si i e mode) m/z:642.148 [M+Na] + , 1261.309 [2M+Na] + . 4.13. 5(6)-Me hyl-2-(2 0 ,3 0 ,4 0 ,6 0 - e a-O-benzoyl-b- D - glucopy anosyl)-benzimidazole (8 ) 490 A: F om acid 5(0.1 g, 0.16 mmol) and 3,4-diamino oluene ( , 0.02 g, 0.16 mmol) acco ding o Gene al p ocedu e I. Yield: 0.06 g (53%). B: F om imida e 6(0.10 g, 0.15 mmol) and 3,4-diamino oluene ( , 0.04 g, 0.30 mmol) acco ding o Gene al P ocedu e II. Reac ion ime: 2 h. Pu ified by column ch oma og aphy (2:3 E OAc–hexane) o yield 0.09 g (83%) o pale yellow sy up. R : 0.41 (1:1 E OAc–hex- ane); [ a ] D =65 (c0.5, CHCl 3 ); 1 H NMR (CDCl 3 )d(ppm): 10.73 (1H, b s, benzimidazole NH), 7.92–6.90 (23H, m, a oma ics), 6.19, 6.11, 5.96 (3 1H, 3 pseudo , J= 9.2, 9.2 Hz in each, H-2 0 , H-3 0 , H-4 0 ), 500 5.32 (1H, d, J= 9.2 Hz, H-1 0 ), 4.68 (1H, dd, J= 12.2, 2.6 Hz, H-6 0 a), 4.54 (1H, dd, J= 12.2, 5.3 Hz, H-6 0 b), 4.44 (1H, ddd, J= 9.2, 5.3, 2.6 Hz, H-5 0 ), 2.32 (3H, s, CH 3 ); 13 C NMR (CDCl 3 )d(ppm): 166.4, 165.9, 165.2 (2) (CO), 148.1 (benzimidazole C-2), 142.4, 138.8 (2 b s, benzimidazole C-3a, C-7a), 133.4–128.0 (a oma ics, benzimid- azole C-5), 124.4, 118.9, 111.1 (b s o each, benzimidazole C-4, C-6, C-7), 76.8, (C-1 0 ), 75.3, 74.1, 71.5, 69.5 (C-1 0 –C-5 0 ), 63.4 (C-6 0 ), 21.5 (CH 3 ). Anal. Calcd o C 42 H 34 N 2 O 9 (710.73): C, 70.98; H, 4.82; N, 3.94. Found: C, 70.83; H, 4.96; N, 4.09. 4.14. 4(7)-Me hyl-2-(2 0 ,3 0 ,4 0 ,6 0 - e a-O-benzoyl-b- D - 510 glucopy anosyl)-benzimidazole (8g) F om imida e 6(0.10 g, 0.15 mmol) and 2,3-diamino oluene (g, 0.04 g, 0.30 mmol) acco ding o Gene al P ocedu e II. Reac ion ime: 5 h. Pu ified by column ch oma og aphy (1:2 E OAc–hexane) o yield 0.087 g (79%) o yellow solid. Mp: 117–119 °C; [ a ] D =43 (c0.5, CHCl 3 ); 1 H NMR (CDCl 3 )d(ppm): 10.88 (1H, b s, benzimid- azole NH), 7.96–6.88 (23H, m, a oma ics), 6.18, 6.06, 5.92 (3 1H, 3 pseudo , J= 9.2, 9.2 Hz in each, H-2 0 , H-3 0 , H-4 0 ), 5.38 (1H, d, J= 9.2 Hz, H-1 0 ), 4.69 (1H, dd, J= 11.7, <1 Hz, H-6 0 a), 4.51 (1H, dd, J= 11.7, 5.3 Hz, H-6 0 b), 4.44 (1H, ddd, J= 9.2, 5.3, <1 Hz, H-5 0 ), 520 2.33 (3H, s, CH 3 ); 13 C NMR (CDCl 3 )d(ppm): 166.3, 165.8, 165.3, 165.2 (CO), 147.7 (benzimidazole C-2), 142.1 (b s, benzimidazole C-3a, C-7a), 133.4–128.0 (a oma ics), 123.1, 122.4, 116.8, 108.8 (b s o each, benzimidazole C-4–C-7), 76.8, 75.2, 73.9, 71.4, 69.5 (C-1 0 –C-5 0 ), 63.3 (C-6 0 ), 16.5 (CH 3 ). Anal. Calcd o C 42 H 34 N 2 O 9 (710.73): C, 70.98; H, 4.82; N, 3.94. Found: C, 70.87; H, 4.90; N, 3.85. 4.15. 5,6-Dime hyl-2-(2 0 ,3 0 ,4 0 ,6 0 - e a-O-benzoyl-b- D - glucopy anosyl)-benzimidazole (8h) A: F om acid 5(0.1 g, 0.16 mmol) and 1,2-diamino-4,5-dime h- 530 ylbenzene (h, 0.02 g, 0.16 mmol) acco ding o Gene al p ocedu e I. Yield: 0.05 g (43%). B: F om imida e 6(0.10 g, 0.15 mmol) and 1,2-diamino-4,5- dime hylbenzene (h, 0.04 g, 0.30 mmol) acco ding o Gene al p o- cedu e II. Reac ion ime: 3 h. Pu ified by column ch oma og aphy (2:3 E OAc–hexane) o yield 0.09 g (81%) o pale yellow solid. Mp: 207–209 °C; [ a ] D =64 (c0.5, CHCl 3 ); 1 H NMR (CDCl 3 )d (ppm): 11.00 (1H, b s, benzimidazole NH), 7.98–6.92 (22H, m, a o- ma ics), 6.27, 6.21, 6.05 (3 1H, 3 pseudo , J= 9.2, 9.2 Hz in each, H-2 0 , H-3 0 , H-4 0 ), 5.34 (1H, d, J= 9.2 Hz, H-1 0 ), 4.67 (1H, dd, J= 11.9, 540 2.6 Hz, H-6 0 a), 4.54 (1H, dd, J= 11.9, 5.3 Hz, H-6 0 b), 4.48 (1H, ddd, J= 9.2, 5.3, 2.6 Hz, H-5 0 ), 2.17 (6H, s, CH 3 ); 13 C NMR (CDCl 3 )d (ppm): 166.3, 165.9, 165.1, 165.0 (CO), 147.5 (benzimidazole C-2), 141.1 (b s, benzimidazole C-3a, C-7a), 133.2–127.9 (a oma - ics, benzimidazole C-5, C-6), 119.5, 111.6 (2 b s, benzimidazole C- 4, C-7), 76.8, 75.6, 74.3, 71.7, 69.5 (C-1 0 –C-5 0 ), 63.3 (C-6 0 ), 20.2 (2 CH 3 ). Anal: Calcd o C 43 H 36 N 2 O 9 (724.75): C, 71.26; H, 5.01; N, 3.87. Found: C, 71.15; H, 4.92; N, 3.99. 4.16. 2-(2 0 ,3 0 ,4 0 ,6 0 -Te a-O-benzoyl-b- D -glucopy anosyl)- naph ho[2,3-d]imidazole (8i) 550 F om imida e 6(0.10 g, 0.15 mmol) and 2,3-diamino-naph ha- lene (i, 0.05 g, 0.30 mmol) acco ding o Gene al p ocedu e II. 6É a Boko e al. /Ca bohyd a e Resea ch xxx (2013) xxx–xxx CAR 6384 No. o Pages 9, Model 5G 5 Feb ua y 2013 Please ci e his a icle in p ess as: Boko ,É.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.01.011 Reac ion ime: 4 h. Pu ified by column ch oma og aphy (4:5 E OAc–hexane) o yield 0.091 g (79%) o pale b own solid. Mp: 184–186 °C; [ a ] D =113 (c0.25, CHCl 3 ); 1 H NMR (CDCl 3 )d (ppm): 10.64 (1H, b s, naph hoimidazole NH), 7.81–6.92 (26H, m, a oma ics), 6.14, 6.05, 5.90 (3 1H, 3 pseudo , J= 9.6, 9.6 Hz in each, H-2 0 , H-3 0 , H-4 0 ), 5.29 (1H, d, J= 9.6 Hz, H-1 0 ), 4.64 (1H, dd, J= 12.3, <1 Hz, H-6 0 a), 4.49 (1H, dd, J= 12.3, 5.3 Hz, H-6 0 b), 4.39 (1H, ddd, J= 9.6, 5.3, <1 Hz, H-5 0 ); 13 C NMR (CDCl 3 )d(ppm): 560 166.2, 165.9, 165.1 (2) (CO), 152.9 (naph hoimidazole C-2), 142.7 (b s, naph hoimidazole C-3a, C-9a), 133.2–127.8, 123.6 (a oma - ics), 116.2, 107.8 (2 b s, naph hoimidazole C-4, C-9), 76.9, 75.6, 74.2, 71.7, 69.5 (C-1 0 –C-5 0 ), 63.4 (C-6 0 ). Analysis: C 45 H 34 N 2 O 9 (746.76), ESI-MS (posi i e mode) m/z:769.219 [M+Na] + , 1516.457 [2M+Na] + . 4.17. 5(6)-Fluo o-2-(b- D -glucopy anosyl)-benzimidazole (10b) F om 8b (0.22 g, 0.30 mmol) acco ding o Gene al P ocedu e III. Reac ion ime: 3.5 h. Pu ified by column ch oma og aphy (4:1 CHCl 3 –MeOH) o yield 0.048 g (54%) o pale yellow sy up. R : 570 0.19 (10:3 CHCl 3 –MeOH); [ a ] D =+9 (c0.2, DMSO); 1 H NMR (CD 3 OD) d(ppm): 7.48 (1H, s, benzimidazole H-4), 7.21 (1H, d, J= 8.7 Hz, benzimidazole H-7), 6.97 (1H, , J= 8.7 Hz, benzimid- azole H-6), 4.46 (1H, d, J= 9.4 Hz, H-1 0 ), 3.88 (1H, dd, J= 11.8, <1 Hz, H-6 0 a), 3.73 (1H, dd, J= 11.8, <1 Hz, H-6 0 b), 3.61 (1H, pseudo , J= 9.8, 9.8 Hz, H-2 0 o H-3 0 o H-4 0 ), 3.52–3.47 (3H, m, H-2 0 and/o H-3 0 and/o H-4 0 , H-5 0 ); 13 C NMR (DMSO-d 6 )d(ppm): 158.3 (d, 1 J C-F = 236 Hz, benzimidazole C-5,), 153.9, 143.2, 138.9, 133.7, 130.6, 119.7, 112.0, 109.7, 104.0, 98.0 (b s o each, benzimid- azole), 81.4, 77.8, 75.9, 72.7, 70.0 (C-1 0 –C-5 0 ), 61.3 (C-6 0 ). Anal. 580 Calcd o C 13 H 15 FN 2 O 5 (298.27): C, 52.35; H, 5.07; N, 9.39. Found: C, 52.26; H, 5.21; N, 9.33. 4.18. 5(6)-Chlo o-2-(b- D -glucopy anosyl)-benzimidazole (10c) F om 8c (0.24 g, 0.33 mmol) acco ding o gene al p ocedu e III. Reac ion ime: 5 h. Pu ified by column ch oma og aphy (4:1 CHCl 3 –MeOH) o yield 0.092 g (89%) o colou less sy up. R : 0.23 (4:1 CHCl 3 –MeOH); [ a ] D = +23 (c0.5, DMSO); 1 H NMR (CD 3 OD) d (ppm): 7.50 (1H, d, J= 1.4 Hz, benzimidazole H-4), 7.45 (1H, d, J= 8.9 Hz, benzimidazole H-7), 7.15 (1H, dd, J= 8.9, 1.4 Hz, benz- imidazole H-6), 4.47 (1H, d, J= 8.9 Hz, H-1 0 ), 3.88 (1H, dd, J= 11.6, 590 <1 Hz, H-6 0 a), 3.73 (1H, dd, J= 11.6, 4.8 Hz, H-6 0 b), 3.61 (1H, pseudo , J= 8.9, 8.9 Hz, H-2 0 o H-3 0 o H-4 0 ), 3.55–3.45 (3H, m, H-2 0 and/o H-3 0 and/o H-4 0 , H-5 0 ); 13 C NMR (CD 3 OD) d(ppm): 155.2 (benz- imidazole C-2), 140.3, 138.1 (2 b s, benzimidazole C-3a, C-7a), 129.2 (benzimidazole C-5), 124.1, 117.1 (b s), 115.9(b s) (benz- imidazole C-4, C-6, C-7), 82.2, 79.2, 77.3, 74.8, 71.2 (C-1 0 –C-5 0 ), 62.8 (C-6 0 ). Anal. Calcd o C 13 H 15 ClN 2 O 5 (314.72): C, 49.61; H, 4.80; N, 8.90. Found: C, 49.72; H, 4.87; N, 8.75. 4.19. 5(6)-B omo-2-(b- D -glucopy anosyl)-benzimidazole (10d) F om 8d (0.27 g, 0.35 mmol) acco ding o Gene al p ocedu e III. 600 Reac ion ime: 6 h. Pu ified by column ch oma og aphy (8:1 CHCl 3 –MeOH) o yield 0.073 g (58%) o colou less sy up. R : 0.23 (10:3 CHCl 3 –MeOH); [ a ] D = +21 (c0.43, DMSO); 1 H NMR (CD 3 OD) d(ppm): 7.65 (1H, s, benzimidazole H-4), 7.41, 7.29 (2 1H, 2d, J= 8.4 Hz, benzimidazole H-6, H-7), 4.47 (1H, d, J= 9.4 Hz, H-1 0 ), 3.88 (1H, dd, J= 11.8, <1 Hz, H-6 0 a), 3.74 (1H, dd, J= 11.8, <1 Hz, H-6 0 b), 3.61 (1H, pseudo , J= 9.4, 8.4 Hz, H-2 0 o H-3 0 o H-4 0 ), 3.53–3.46 (3H, m, H-2 0 and/o H-3 0 and/o H-4 0 , H-5 0 ); 13 C NMR (DMSO-d 6 )d(ppm): 153.8 (benzimidazole C-2), 141.5, 134.8 (2 b s, C-3a, C-7a), 124.5, 121.0, 113.9, 113.8 (b s o each, benzimid- 610 azole C-4–C-7), 81.5, 77.8, 75.9, 72.8, 70.0 (C-1 0 –C-5 0 ), 61.3 (C-6 0 ). Anal. Calcd o C 13 H 15 B N 2 O 5 (359.17): C, 43.47; H, 4.21; N, 7.80. Found: C, 43.43; H, 4.34; N, 7.93. 4.20. 2-(b- D -Glucopy anosyl)-5(6)-ni o-benzimidazole (10e) F om 8e (0.30 g, 0.40 mmol) acco ding o Gene al p ocedu e III. Reac ion ime: 1 day. Pu ified by column ch oma og aphy (85:15 CHCl 3 –MeOH) o yield 0.07 g (53%) o yellow sy up. R : 0.27 (4:1 CHCl 3 –MeOH); 1 H NMR (DMSO-d 6 )d(ppm): 13.03 (1H, b s, benz- imidazole NH), 8.44 (1H, d, J= 1.3 Hz, benzimidazole H-4), 8.11 (1H, dd, J= 9.2, 1.3 Hz, benzimidazole H-6), 7.70 (1H, d, J= 9.2 Hz, 620 benzimidazole H-7), 5.14 (3H, b s, 3 OH), 4.60 (1H, b s, OH), 4.42 (1H, d, J= 9.2 Hz, H-1 0 ), 3.74 (1H, dd, J= 11.9, <1 Hz, H-6 0 a), 3.63 (1H, pseudo , J= 9.2, 9.2 Hz, H-2 0 o H-3 0 o H-4 0 ), 3.49 (1H, dd, J= 11.9, 5.3 Hz, H-6 0 b), 3.39–3.34 (2H, m, H-2 0 o H-3 0 o H-4 0 , H-5 0 ), 3.24 (1H, pseudo , J= 9.2, 9.2 Hz, H-2 0 o H-3 0 o H-4 0 ); 13 C NMR (DMSO-d 6 )d(ppm): 157.3 (benzimidazole C-2), 142.5 (benz- imidazole C-5), 141.4, 138.8 (2 b s, benzimidazole C-3a, C-7a), 117.7, 114.3 (b s), 112.6 (b s) (benzimidazole C-4, C-6, C-7), 81.6, 77.7, 76.0, 72.8, 70.0 (C-1 0 –C-5 0 ), 61.2 (C-6 0 ). Anal. Calcd o C 13 H 15 N 3 O 7 (325.27): C, 48.00; H, 4.65; N, 12.92. Found: C, 47.87; 630 H, 4.56; N, 12.96. 4.21. 2-(b- D -Glucopy anosyl)-5(6)-me hyl-benzimidazole (10 ) F om 8e (0.27 g, 0.38 mmol) acco ding o Gene al p ocedu e III. Reac ion ime: 3 h. Pu ified by column ch oma og aphy (85:15 CHCl 3 –MeOH) o yield 0.10 g (87%) o colou less sy up. R : 0.42 (7:3 CHCl 3 –MeOH); [ a ] D = +18 (c0.5, MeOH); 1 H NMR (CD 3 OD) d (ppm): 7.41 (1H, d, J= 7.9 Hz, benzimidazole H-6 o H-7), 7.32 (1H, s, benzimidazole H-4), 7.03 (1H, d, J= 7.9 Hz, benzimidazole H-6 o H-7), 4.49 (1H, d, J= 9.2 Hz, H-1 0 ), 3.91 (1H, dd, J= 11.9, <1 Hz, H-6 0 a), 3.76 (1H, dd, J= 11.9, 4.0 Hz, H-6 0 b), 3.66 (1H, pseudo 640 , J= 9.2, 9.2 Hz, H-2 0 o H-3 0 o H-4 0 ), 3.61–3.49 (3H, m, H-2 0 and/o H-3 0 and/o H-4 0 , H-5 0 ), 2.42 (3H, s, CH 3 ); 13 C NMR (CD 3 OD) d (ppm): 153.4 (benzimidazole C-2), 139.0, 137.6 (benzimidazole C-3a, C-7a), 133.6 (benzimidazole C-5), 125.2, 115.9, 115.4 (benz- imidazole C-4, C-6, C-7), 82.1, 79.3, 77.3, 74.8, 71.2 (C-1 0 –C-5 0 ), 62.7 (C-6 0 ), 21.7 (CH 3 ). Anal. Calcd o C 14 H 18 N 2 O 5 (294.30): C, 57.13; H, 6.16; N, 9.52. Found: C, 57.29; H, 6.07; N, 9.39. 4.22. 2-(b- D -Glucopy anosyl)-4(7)-me hyl-benzimidazole (10g) F om 8g (0.25 g, 0.35 mmol) acco ding o Gene al P ocedu e III. Reac ion ime: 4 h. Pu ified by column ch oma og aphy (4:1 650 CHCl 3 –MeOH) o gi e 0.075 g (73%) o colou less sy up. R : 0.20 (4:1 CHCl 3 –MeOH); [ a ] D = +13 (c0.5, DMSO); 1 H NMR (DMSO-d 6 ) d(ppm): 12.46 (1H, b s, benzimidazole NH), 7.32, 7.04, 6.95 (3 1H, benzimidazole), 5.14 (3H, b s, 3 OH), 4.64 (1H, b s, OH), 4.36 (1H, d, J= 10.2 Hz, H-1 0 ), 3.73–3.24 (6H, m, H-2 0 , H-3 0 , H-4 0 , H-5 0 , H-6 0 a, H-6 0 b), 2.50 (3H, s, CH 3 ); 13 C NMR (DMSO-d 6 )d (ppm): 151.9 (benzimidazole C-2), 141.8, 133.5, 128.4, 121.6, 115.6, 109.0 (b s o each, benzimidazole C-3a, C-7a, C-4–C-7), 81.3, 77.9, 75.9, 72.7, 70.0 (C-1 0 –C-5 0 ), 61.2 (C-6 0 ). Anal. Calcd o C 14 H 18 N 2 O 5 (294.30): C, 57.13; H, 6.16; N, 9.52. Found: C, 57.18; 660 H, 6.05; N, 9.64. 4.23. 5,6-Dime hyl-2-(b- D -glucopy anosyl)-benzimidazole (10h) F om 8h (0.27 g, 0.37 mmol) acco ding o Gene al p ocedu e III. Reac ion ime: 3 h. Pu ified by column ch oma og aphy (85:15 CHCl 3 –MeOH) o yield 0.09 g (78%) o colou less sy up. R : 0.42 (7:3 CHCl 3 –MeOH); [ a ] D = +24 (c0.5, MeOH); 1 H NMR (CD 3 OD) d (ppm): 7.28 (2H, s, benzimidazole H-4, H-7), 4.47 (1H, d, J= 9.2 Hz, H-1 0 ), 3.91 (1H, dd, J= 11.9, <1 Hz, H-6 0 a), 3.76 (1H, dd, É a Boko e al. /Ca bohyd a e Resea ch xxx (2013) xxx–xxx 7 CAR 6384 No. o Pages 9, Model 5G 5 Feb ua y 2013 Please ci e his a icle in p ess as: Boko ,É.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.01.011