Synthesis of some O-, S- and N-glycosides of hept-2-ulopyranosonamides

UNCORRECTED PROOF Note Synthesis of some O-, Sand N-glycosides of hept-2-ulopyranosonamides Veronika Nagy a , Katalin Czifrák a , Attila Bényei b , László Somsák a,* a Department of Organic Chemistry (POB 20), University of Debrecen, H-4010 Debrecen, Hungary b Department of Physical Chemistry (POB 7), University of Debrecen, H-4010 Debrecen, Hungary Q2 10 article info Article history: Received 30 December 2008 Received in revised form 6 February 2009 Accepted 7 February 2009 Available online xxxx Keywords: Hept-2-ulopyranosonamides O-glycoside S-glycoside 20 N-glycoside abstract (O-Peracylated a -D-glucoand -galacto-hept-2-ulopyranosylbromide)onamides gave the corresponding (alkyl b-D-glyco-hept-2-ulopyranoside)onamides under Koenigs–Knorr conditions, and similar aryl glycosides were obtained with sodium phenolates; (aryl and hetaryl 2-thio-b-D-gluco-hept-2-ulopyranoside)onamides were formed with thiophenols in the presence of K 2 CO 3 in acetone, and reactions with aniline in CH 2 Cl 2 furnished (N-phenyl b-D-glyco-hept-2-ulopyranosylamine)onamides. Some deprotected derivatives of D-gluco configuration obtained by the Zemplén protocol showed no significant inhibition against rabbit muscle glycogen phosphorylase b. Ó2009 Published by Elsevier Ltd. C-(2,3,4,6-Tetra-O-acyl-1-bromo-1-deoxy-bD -glycopyranosyl)formamides (3,4,5,7-tetra-O-acyla - D -glyco-hept-2-ulopyranosylbromide)onamides) 1–5 (e.g., 1 1,2 and 2 5 ) proved versatile starting materials for the syntheses of diverse monosaccharide derivatives. Thus, their reactions with nucleophiles such as H 2 O, 6 azide ion, 7,8 nitriles, 9 acetone and DMSO, 10 cyanate and thiocyanate ions, 2,3,6 the latter two resulting in cyclisations to give glycopyranosylid40 ene-spiro-(thio)hydantoins efficient glucose analogue glycogen phosphorylase inhibitors (GPIs), 11–15 as well as eliminations to substituted glycals 1 were reported. Several derivatives of D -glucose with a CONH 2 moiety in the a -anomeric position were shown to be GPIs, 16–18 although a clearcut conclusion for the role of this group could not yet be drawn. 8 In order to produce new compounds of this type, and to study the reactivity of hept-2-ulopyranosylonamide bromides towards further nucleophiles, we have investigated the preparation of some O-, Sand N-glycosidic derivatives from 1 and 2. 50 Treatment of 1with MeOH or EtOH as the solvent in the presence of Ag 2 CO 319 gave methyl and ethyl glycosides 3and 4, respectively (Table 1,entries 1 and 2). Decreasing the amount of EtOH in CH 2 Cl 2 as co-solvent was investigated (entries 3–6) to show that as few as 2 equiv of the alcohol gave satisfactory results. Changing the promoter to the more efficient AgOTf significantly reduced the reaction time and increased the yield (entry 6). A large excess of n-BuOH gave the corresponding glycoside 7in satisfactory yield (entry 7). On the other hand, reactions of 1with t-BuOH or BnOH (entries 8 and 9), and similarly, those of 2with EtOH or n-BuOH 60 (entries 12 and 13) gave significant amounts of the corresponding O-peracylated a - D -glyco-hept-2-ulopyranosonamides 15 2 and 16 4,5 besides the expected glycosides 7and 8as well as 12 and 14, respectively. The reaction of 2-nitrophenol with 1in the presence of AgOTf and Et 3 N or DBU gave 9in 32% and 24% yields, respectively. Under phase transfer conditions (2-nitrophenol in CH 2 Cl 2 , 1MNaOH in water, Bu 4 NBr) 9could not be observed in the reaction mixture. Therefore, we turned to the sodium salt of 2-nitrophenol (entry 10), however, this reaction again gave 9in a low yield accompanied by 15. On the contrary, sodium 4-nitropheno70 late (entry 11) gave the expected 10 in good yield. The steric accessibility of the nucleophilic part of the reagents may be responsible for the large differences in the outcomes of these reactions. Deprotection of glycosides 4and 12 was effected by the Zemplén protocol, while 10 was deacetylated by KCN/MeOH to give 5,13 and 11, respectively. For the formation of N-phenyl-glycosylamines, 20,21 1and 2 were reacted with aniline to give 17 and 18, respectively (Scheme 1). The latter was deprotected by the Zemplén method to yield 19. To obtain S-glycosides, 22 2was reacted with thiols in acetone in 80 the presence of K 2 CO 3 to give the expected products 20,22 and 24 in good yields (Scheme 1). For deprotection of these compounds the Zemplén method was applied to give 21,23 and 25, respectively, without difficulties. Structure elucidation of the new compounds was straightforward by NMR methods. The 4 C 1 conformation of the pyranose rings followed from the vicinal proton–proton coupling constants. For most representative compounds, the configuration of the anomeric carbon was established on the basis of three-bond heteronuclear couplings between H-2 (parent sugar numbering) and the 0008-6215/$ - see front matter Ó2009 Published by Elsevier Ltd. doi:10.1016/j.carres.2009.02.011 *Corresponding author. Tel.: +36 52 512 900/22348; fax: +36 52 453 836 Q1 . E-mail address: [email protected] (L. Somsák). Carbohydrate Research xxx (2009) xxx–xxx Contents lists available at ScienceDirect Carbohydrate Research journal homepage: www.elsevier.com/locate/carres CAR 4933 No. of Pages 7, Model 5G 20 February 2009 Disk Used ARTICLE IN PRESS Please cite this article in press as: Nagy, V. et al., Carbohydr. Res. (2009), doi:10.1016/j.carres.2009.02.011 UNCORRECTED PROOF 90 exocyclic carbonyl of the amide group measured as earlier. 8 The values larger than 4 Hz suggested trans arrangement of the relevant atoms in the 4 C 1 conformation. 8 In case of 4, a single crystal X-ray structure determination unequivocally confirmed the anomeric configuration (Fig. 1). The investigated substitution reactions were clean, that is, disregarding by-products 15 and 16 no other compounds than the isolated products were observed by TLC. This reveals exclusive stereoselectivity for each transformation. An explanation for this can be an S N 2 type replacement of bromine in the cases of pheno100 lates, aniline and thiolates. In the reactions with alcohols promoted by silver salts neighbouring group participation of the 2-acyloxy substituent in the possible intermediate glycosylium ion may account for the inversion. However, given the electron-withdrawing character of the carboxamido group, formation of the glycosylium ion may be unfavourable. Therefore, an electrophilically assisted Table 1 Preparation of (alkyl or aryl b-D-glyco-hept-2-ulopyranoside)onamides O R R 1 R R OR' CONH 2 O R R 2 R R CONH 2 Br R'OH or R'ONa O R R 2 R R CONH 2 + promoter, solvent, OH R 1 R 2 R 1 1 R = R 1 = OAc, R 2 = H 2 R = R 2 = OBz, R 1 = H 12, 14 R = R 2 = OBz, R 1 = H 3, 4, 6-10 R = R 1 = OAc, R 2 = H 4 5 R = R 1 = OH, R 2 = H, R' = Et (98%) 10 11 R = R 1 = OH, R 2 = H, R' = 4-NO 2 -C 6 H 4 (99%) 12 13 R = R 2 = OH, R 1 = H, R' = Et (98%) i NaOMe, MeOH, rt ii KCN, MeOH, rt i ii i rt 15 R = R 1 = OAc, R 2 = H 16 R = R 2 = OBz, R 1 = H Entry Starting compound R 0 OH or R 0 ONa (equiv) Promoter Solvent Reaction time Product(s) (Yield [%]) 11MeOH (as solvent) Ag 2 CO 3 MeOH 2 h 3(89) — 21EtOH (as solvent) Ag 2 CO 3 EtOH 2 h 4(85) — 31EtOH (70) Ag 2 CO 3 CH 2 Cl 2 2h 4(84) — 41EtOH (10) Ag 2 CO 3 CH 2 Cl 2 1d 4(93) — 51EtOH (2) Ag 2 CO 3 CH 2 Cl 2 2d 4(50) — 61EtOH (2) AgOTf CH 2 Cl 2 5 min 4(80) — 71n-BuOH (70) Ag 2 CO 3 CH 2 Cl 2 2d 6(90) — 81t-BuOH (10) Ag 2 CO 3 CH 2 Cl 2 7d 7(21) 15 (29) 91C 6 H 5 CH 2 OH (10) Ag 2 CO 3 CH 2 Cl 2 7d 8(31) 15 (9) 10 1 ONa NO2 (5) —CH 2 Cl 2 36 d 9(24) 15 (25) 11 1 ONa O2N (5) —CH 3 CN 1 d 10 (82) — 12 2EtOH (50) Ag 2 CO 3 CH 2 Cl 2 2d 12 (87) 16 (10) 13 2n-BuOH (43) Ag 2 CO 3 CH 2 Cl 2 6d 14 (56) 16 (31) O R R2 RR NHPh CONH2 O R R2 RR CONH2 Br PhNH2, CH2Cl2, rt 1 R = R1 = OAc, R2 = H 2 R = R2 = OBz, R1 = H 17 R = R1 = OAc, R2 = H (75%) 18 R = R2 = OBz, R1 = H (55%) 19 R = R2 = OH, R1 = H (54%) i NaOMe, MeOH, rt R'SH, K2CO3, acetone, rt from 2 i R1 R1 O OR RO RO RO SR' CONH 2 N S N 20 (79%) 21 (86%) 22 (73%) 23 (62%) 24 (76%) 25 (47%) i i i R' R = Bz R = H Scheme 1. 2V. Nagy et al. / Carbohydrate Research xxx (2009) xxx–xxx CAR 4933 No. of Pages 7, Model 5G 20 February 2009 Disk Used ARTICLE IN PRESS Please cite this article in press as: Nagy, V. et al., Carbohydr. Res. (2009), doi:10.1016/j.carres.2009.02.011 UNCORRECTED PROOF substitution of bimolecular character can also be taken into consideration. The deprotected compounds were assayed against rabbit muscle glycogen phosphorylase b as described earlier, 3,6 and showed 110 no significant inhibition (13 21% at 625 l M; 19 IC 50 > 60 mM; 21, 23,25 no inhibition at 625 l M). 1. Experimental 1.1. General methods Melting points were measured in open capillary tubes or on a Kofler hot-stage and are uncorrected. Optical rotations were determined with a Perkin–Elmer 241 polarimeter at rt. NMR spectra were recorded with Bruker 200 (200/50 MHz for 1 H/ 13 C), Bruker 360 (360/90 MHz for 1 H/ 13 C) or Avance DRX 500 (500/125 MHz for 1 H/ 13 C) spectrometers. Chemical shifts are referenced to Me 4 Si 120 ( 1 H) or to the residual solvent signals ( 13 C). TLC was performed on DC-Alurolle Kieselgel 60 F 254 (Merck), and the plates were visualised under UV light and by gentle heating. For column chromatography, Kieselgel 60 (Merck, particle size 0.063–0.200 mm) was used. Dichloromethane was distilled from P 4 O 10 and acetone from CaSO 4 ) and stored over 4Å molecular sieves. Organic solutions were dried over anhydrous MgSO 4 and were concentrated under diminished pressure at 40–50 °C (water bath). 1.2. General procedure I for the preparation of C-(2,3,4,6-tetra-Oacetyl-1-alkoxya - D -glycopyranosyl)formamides ((alkyl 3,4,5,7130 tetra-O-acetyl-bD -galacto-hept-2-ulopyranoside)onamides) To a solution of a C-(2,3,4,6-tetra-O-acyl-1-bromo-1-deoxy-bD - glycopyranosyl)formamide, ((3,4,5,7-tetra-O-acyla - D -glyco-hept2-ulopyranosylbromide)onamide) (1 1,2 or 2, 5 0.3 mmol) in dry CH 2 Cl 2 (2 mL) containing molecular sieves (0.1 g, 3 Å) an alcohol (Table 1) and Ag 2 CO 3 (1 equiv, 0.30 mmol, 0.08 g) or silver triflate (1 equiv, 0.30 mmol, 0.07 g) and Et 3 N(1equiv, 0.30 mmol, 39 l L) were added. The reaction mixture was stirred in the dark at rt until TLC (1:1 EtOAc–hexane) showed the complete transformation of the starting material. The mixture was then filtered on a Celite 140 pad, and the solvent was removed under diminished pressure. The crude product was crystallised from EtOAc–hexane or purified by column chromatography. 1.3. General procedure II for the preparation of C-(2,3,4,6-tetra-O -acyl-1-deoxy-1-phenylaminoa - D -glycopyranosyl)formamides ((N-Phenyl 3,4,5,7-tetra-O-acyl-bD -glyco-hept-2-ulopyranosylamine)onamides) To a solution of C-(2,3,4,6-tetra-O-acyl-1-bromo-1-deoxy-bD - glycopyranosyl)formamide, ((3,4,5,7-tetra-O-acyla - D -glyco-hept2-ulopyranosylbromide)onamide) (1 1,2 or 2, 5 0.3 mmol) in dry 150 CH 2 Cl 2 (2 mL) containing molecular sieves (0.1 g, 3 Å), aniline (50 equiv to 1and 5 equiv to 2) was added. The reaction mixture was stirred at rt until TLC (1:1 EtOAc–hexane) showed the complete transformation of the starting sugar (1–2 d). The mixture was then filtered on a Celite pad and the solvent was evaporated. The residue was dissolved in EtOAc, the solution was washed with water, diluted hydrochloric acid, and satd aq NaHCO 3 solution. After drying and solvent removal the crude product was crystallised from EtOAc–hexane or purified by column chromatography. 1.4. General procedure III for the preparation of C-(2,3,4,6160 tetra-O-benzoyl-1-deoxy-1-aryl or heteroarylsulfanyla - D - glucopyranosyl)formamides ((arylor heteroaryl3,4,5,7-tetraO-benzoyl-2-thio-bD -gluco-hept-2-ulopyranoside)onamides) To a solution of C-(1-bromo-1-deoxy-2,3,4,6-tetra-O-benzoyl-bD -glucopyranosyl)-formamide, 5 ((3,4,5,7-tetra-O-benzoyla - D -gluco-hept-2-ulopyranosylbromide)onamide) (2, 0.20 g, 0.28 mmol) in dry acetone (3 mL) containing molecular sieves (0.1 g, 3 Å), a thiol (1.40 mmol) and K 2 CO 3 (0.20 g, 1.40 mmol) were added. The reaction was stirred at rt until TLC (1:2 EtOAc–hexane) showed complete transformation of the starting material. The mixture 170 was then filtered, diluted with CH 2 Cl 2 (5 mL), washed with satd aq NaHCO 3 solution (2 5 mL), and water (1 5 mL). After drying and solvent removal, the crude product was purified by column chromatography. 1.5. General procedure IV for the Zemplén-deacylation To a solution of an O-acyl protected compound in dry MeOH 1–2 drops of a 1 M methanolic NaOMe solution were added, and the reaction mixture was maintained at rt until completion of the transformation TLC (1:1 CHCl 3 –MeOH). Amberlyst 15 (H + form) was then added to remove sodium ions, the resin was fil180 tered off, and the solvent was removed under diminished pressure. If the residue was chromatographically not uniform it was purified by column chromatography or crystallisation. 1.6. C-(2,3,4,6-Tetra-O-acetyl-1-methoxya - D -galactopyranosyl) formamide ((Methyl 3,4,5,7-tetra-O-acetyl-bD -galacto-hept-2ulopyranoside)onamide) (3) This compound was prepared from 1(0.30 g 0.66 mmol) according to General procedure I. The crude product was crystallised to give 3(0.22 g, 85%) as a yellowish crystalline product. Mp: 120–122 °C; [ a ] D +69 (c1.03, CHCl 3 ); 1 H NMR (CDCl 3 , 190 360 MHz): d(ppm) 6.65 (s,1H, NH), 5.93 (s, 1H, NH), 5.86 (dd, 1H, J 2,3 10.5 Hz, J 3,4 3.1 Hz, H-3), 5.53 (d, 1H, J 2,3 10.5 Hz, H-2), 5.50 (dd, 1H, J 3,4 3.1 Hz, J 4,5 1.5 Hz, H-4), 4.85 (ddd, 1H, J 5,6 6.3 Hz, J 5,6 0 5.3 Hz, H-5), 4.12 (dd, 1H, J 6,6 0 11.0 Hz, H-6), 4.05 (dd, 1H, J 6,6 0 11.0 Hz, H-6 0 ), 3.48 (s, 3H, CH 3 ), 2.14, 2.04, 2.01, 1.95 (4 s, 12H, OCOCH 3 ); 13 C NMR (CDCl 3 , 90 MHz): d(ppm): 170.4 (CONH 2 , 3 J H-2,CO =4.7 Hz), 169.7 (2), 169.6 (2) (CO), 97.5 (C-1), 71.1, 70.0, 67.4, 64.7 (C-2 to C-5), 61.5 (C-6), 49.8 (OCH 3 ), 20.7, Figure 1. Ortep view at 40% probability level and partial crystallographic numbering scheme of compound 4. Selected torsion angles (°) for the two molecules in the asymmetric unit: O5–C1–O1–C8: 58 and 46; O1–C1–C7–N1: 3 and 12. V. Nagy et al. / Carbohydrate Research xxx (2009) xxx–xxx 3 CAR 4933 No. of Pages 7, Model 5G 20 February 2009 Disk Used ARTICLE IN PRESS Please cite this article in press as: Nagy, V. et al., Carbohydr. Res. (2009), doi:10.1016/j.carres.2009.02.011 UNCORRECTED PROOF 20.6, 20.5 (COCH 3 ). Anal. Calcd for C 16 H 23 NO 11 (405.36): C, 47.41; H, 5.72; N, 3.46. Found: C, 47.00; H, 5.52; N, 3.23. 200 1.7. C-(2,3,4,6-Tetra-O-acetyl-1-ethoxya - D -galactopyranosyl) formamide ((ethyl 3,4,5,7-tetra-O-acetyl-bD -galacto-hept-2ulopyranoside)onamide) (4) This compound was prepared from 1(0.20 g 0.44 mmol) according to General procedure I. The crude product was crystallised from hexane to give 4(0.16 g, 85%) as a white crystalline product. Mp 135–137 °C; [ a ] D +57 (c1.04, CHCl 3 ); 1 H NMR (CDCl 3 , 360 MHz): d(ppm) 6.69 (s, 1H, NH), 6.26 (s, 1H, NH), 5.85 (dd, 1H, J 2,3 10.3 Hz, J 3,4 3.6 Hz, H-3), 5.51 (d, 1H, J 2,3 10.3 Hz, H-2), 5.46 (dd, 1H, J 3,4 3.6 Hz, J 4,5 1.2 Hz, H-4), 4.81 (pseudo t, 1H, J 5,6 6.8 Hz, J 5,6 0 210 6.7 Hz, H-5), 4.10–3.99 (m, 2H, H-6, H-6 0 ), 3.88–3.70 (m, 2H, CH 2 ), 2.12, 2.05, 1.99, 1.91 (4 s, 12H, OCOCH 3 ), 1.19 (m, 3H, CH 3 ), 13 C NMR (CDCl 3 , 90 MHz): d(ppm) 170.3 (CONH 23 J H2,CO =6.1 Hz), 169.9 (2), 169.7 (2) (CO), 97.4 (C-1), 70.9, 70.0, 67.4, 65.4 (C-2 to C-5), 61.4 (C-6). 58.3 (CH 2 ), 20.7, 20.6, 20.5 (2) (COCH 3 ), 15.3 (CH 3 ). Anal. Calcd for C 17 H 25 NO 11 (419.10): C, 48.69; H, 6.01; N, 3.34. Found: C, 48.54; H, 6.04; N, 3.49. 1.8. C-(1-Ethoxya - D -galactopyranosyl)formamide ((ethyl bD - galacto-hept-2-ulopyranoside)onamide) (5) This compound was prepared from 4(0.20 g 0.47 mmol) 220 according to General procedure IV to give 5(0.12 g, 98%) as a yellowish oil. R f = 0.32 (7:3 CHCl 3 –MeOH); [ a ] D +61 (c1.26, H 2 O); 1 H NMR (D 2 O, 360 MHz): d(ppm) 4.28–3.55 (m, 8H, H-2, H-3, H-4, H5, H-6, H-6 0 ,CH 2 ), 1.18 (t, 3H, J7.3 Hz, CH 3 ), 13 C NMR (D 2 O, 90 MHz): d(ppm) 173.7 (CONH 2 , 3 J H-2,CO =4.8 Hz), 100.6 (C-1), 76.4, 72.8, 71.0, 70.1 (C-2 to C-5), 62.7 (C-6), 59.4 (CH 2 ), 15.9 (CH 3 ). Anal. Calcd for C 9 H 17 NO 7 (251.24): C, 43.03; H, 6.82; N, 5.58. Found: C, 43.54; H, 6.63; N, 5.12. 1.9. C-(2,3,4,6-Tetra-O-acetyl-1-n-buthoxya - D -galactopyranosyl)formamide ((n-buthyl 3,4,5,7-tetra-O-acetyl-bD -galacto230 hept-2-ulopyranoside)onamide) (6) This compound was prepared from 1(0.20 g 0.44 mmol) according to General procedure I. The crude product was crystallised from hexane to give 6(0.18 g, 90%) as a white crystalline product. Mp: 97–99 °C; [ a ] D +48 (c1.24, CHCl 3 ); 1 H NMR (CDCl 3 , 360 MHz): d(ppm) 6.65 (s, 1H, NH), 6.42 (s, 1H, NH), 5.84 (dd, 1H, J 2,3 10.5 Hz, J 3,4 3.1 Hz, H-3), 5.50 (d, 1H, J 2,3 10.5 Hz, H-2), 5.47 (dd, 1H, J 3,4 3.1 Hz, J 4,5 1.2 Hz, H-4), 4.81 (pseudo t, 1H, J 5,6 5.8 Hz, J 5,6 0 .8 Hz, H-5), 4.15–3.99 (m, 2H, H-6, H-6 0 ), 3.80–3.64 (m, 2H, CH 2 ), 2.11, 2.01, 1.98, 1.91 (4 s, 12H, OCOCH 3 ), 1.60–1.55 240 (m, 2H, CH 2 ), 1.38–1.32 (m, 2H, CH 2 ), 0.91 (t, 3H, J6.8 Hz, CH 3 ); 13 C NMR (CDCl 3 , 90 MHz): d(ppm) 170.3, 169.9, 169.7, 169.6 (CO), 170.0 (CONH 2 , 3 J H-2,CO =6.1 Hz), 97.3 (C-1), 70.9, 70.0, 67.3, 65.4 (C-2 to C-5), 62.1 (C-6), 61.3, 31.5 (CH 2 ), 20.6, 20.5, 20.5 (COCH 3 ) 19.0 (CH 2 ), 13.6 (CH 3 ). Anal. Calcd for C 19 H 29 NO 11 (447.44): C, 51.00; H, 6.53; N, 3.13. Found: C, 51.15; H, 6.57; N, 3.29. 1.10. C-(2,3,4,6-Tetra-O-acetyl-1-t-buthoxya - D -galactopyranosyl)formamide ((t-buthyl 3,4,5,7-tetra-O-acetyl-bD -galactohept-2-ulopyranoside)onamide) (7) 250 This compound was prepared from Q3 1(0.20 g 0.44 mmol) according to General procedure I, and was purified by column chromatography (1:1 EtOAc–hexane) to give 7(0.04 g, 21%) as a colourless oil, and in the second fraction it gave compound 15 (29%). R f = 0.42 (3:1 EtOAc–hexane); [ a ] D +19 (c1.02, CHCl 3 ); 1 H NMR (CDCl 3 , 360 MHz): d(ppm) 6.81 (s, 1H, NH), 6.19 (s, 1H, NH), 5.84 (dd, 1H, J 2,3 10.5 Hz, J 3,4 3.1 Hz, H-3), 5.60 (d, 1H, J 2,3 10.5 Hz, H-2), 5.50 (dd, 1H, J 3,4 3.1 Hz, J 4,5 1.5 Hz, H-4), 4.91 (pseudo t, 1H, J 5,6 6.3 Hz, J 5,6 0 6.3 Hz, H-5), 4.13–4.10 (m, 2H, H-6, H-6 0 ), 2.10, 2.02, 2.00, 1.93 (4 s, 12H, OCOCH 3 ), 1.42 (s, 9H, C(CH 3 ) 3 ); 260 13 C NMR (CDCl 3 , 90 MHz): d(ppm) 171.4 (CONH 2 , 3 J H-2,CO =5.8 Hz), 170.3, 169.8, 169.7 (2) (CO), 98.7 (C(CH 3 ) 3 ), 80.1 (C-1), 71.3, 70.0, 96.9, 67.5 (C-2 to C-5), 61.4 (C-6), 30.1 (C(CH 3 ) 3 ), 20.8, 20.6 (3) (COCH 3 ). Anal. Calcd for C 19 H 29 NO 11 (447.44): C, 51.00; H, 6.53; N, 3.13. Found: C, 51.17; H, 6.52; N, 3.30. 1.11. C-(2,3,4,6-Tetra-O-acetyl-1-benzyloxya - D -galactopyranosyl)formamide ((benzyl 3,4,5,7-tetra-O-acetyl-bD -galactohept-2-ulopyranoside)onamide) (8) This compound was prepared from 1(0.20 g 0.44 mmol) 270 according to General procedure I, and was purified by column chromatography (1:1 EtOAc–hexane) to give 8(0.07 g, 31%) as a white crystalline product, and in the second fraction it gave compound 15 (9%). Mp 163–164 °C; [ a ] D +16 (c1.03, CHCl 3 ); 1 HNMR (CDCl 3 , 360 MHz): d(ppm) 7.42–7.33 (m, 5H, ArH), 6.63 (s, 1H, NH), 5.93 (dd, 1H, J 2,3 10.3 Hz, J 3,4 3.1 Hz, H-3), 5.68 (d, 1H, J 2,3 10.3 Hz, H-2), 5.58–5.54 (m, 2H, H-4, NH), 4.93 (pseudo t, 1H, J 5,6 6.8 Hz, J 5,6 0 6.6 Hz, H-5) 4.90, 4.74 (2 d, 2H, J10.5 Hz, CH 2 ), 4.18–4.12 (m, 2H, H-6, H-6 0 ), 2.17, 2.08, 2.03, 1.98 (4 s, 12H, OCOCH 3 ), 13 C NMR (CDCl 3 , 90 MHz): d(ppm) 170.3 (CONH 2 , 280 3 J H-2,CO =6.1 Hz), 169.9, 169.8 (2), 169.7 (CO), 136.7, 128.5, 128.4, 128.0 (ArC), 97.5 (C-1), 71.2, 69.9, 67.3, 65.7 (C-2 to C-5), 64.7 (CH 2 ),61.3 (C-6), 20.7, 20.5, 20.5 (3) (COCH 3 ). Anal. Calcd for C 22 H 27 NO 11 (481.46): C, 54.88; H, 5.65; N, 2.91. Found: C, 54.09; H, 5.62; N, 2.92. 1.12. C-[2,3,4,6-Tetra-O-acetyl-1-(2-nitrophenoxy)- a - D -galactopyranosyl]formamide ((2-nitrophenyl 3,4,5,7-tetra-O-acetyl-bD -galacto-hept-2-ulopyranoside)onamide) (9) To a solution of 1(0.20 g, 0.44 mmol) in dry CH 2 Cl 2 (2 mL) containing molecular sieves (3 Å), sodium 2-nitrophenolate (0.35 g, 290 2.20 mmol) was added. The reaction mixture was stirred at rt until TLC (1:1 EtOAc–hexane) showed complete transformation of the starting sugar (36 d). Then the mixture was filtered on a Celite pad, and the solvent was removed. The oily residue was purified by column chromatography (1:1 EtOAc–hexane) to give 9(0.04 g, 24%) as a yellow oil, and in the second fraction it gave compound 15 (0.04 g, 25%). Characterisation of 9:R f = 0.72 (1:3 EtOAc–hexane); [ a ] D +52 (c0.49, CHCl 3 ); 1 H NMR (CDCl 3 , 360 MHz): d (ppm) 7.97–7.85 (m, 2H, ArH), 7.56 (m, 1H, ArH) 7.28 (s, 1H, NH), 7.26–7.23 (m, 1H, ArH), 6.23 (s, 1H, NH), 5.79 (dd, 1H, J 2,3 300 10.2 Hz, J 3,4 3.1 Hz, H-3), 5.62 (d, 1H, J 2,3 10.2 Hz, H-2), 5.54 (dd, 1H, J 3,4 3.1 Hz, J 4,5 1.1 Hz, H-4), 5.19 (pseudo t, 1H, J 5,6 6.1 Hz, J 5,6 0 6.1 Hz, H-5), 4.11–4.06 (m, 2H, H-6, H-6 0 ), 2.09, 2.03, 2.01, 1.89 (4 s, 12H, OCOCH 3 ); 13 C NMR (CDCl 3 , 90 MHz): d(ppm): 170.3, 169.5, 169.3, 168.6 (2) (CO), 146.1, 134.0, 126.1, 124.3, 121.5 (ArC), 100.7 (C-1), 72.6, 69.9, 67.3, 65.3 (C-2 to C-5), 61.4 (C-6), 20.5, 20.4, 20.3 (2) (COCH 3 ). Anal. Calcd for C 21 H 24 N 2 O 13 (512.43): C, 49.22; H, 4.72; N, 5.47. Found: C, 50.05; H, 4.53; N, 5.29. 1.13. C-[2,3,4,6-Tetra-O-acetyl-1-(4-nitrophenoxy)- a - D -galacto310 pyranosyl]formamide ((4-nitrophenyl 3,4,5,7-tetra-O-acetyl-bD -galacto-hept-2-ulopyranoside)onamide) (10) To a solution of 1(1.0 g, 2.20 mmol) in dry CH 3 CN (10 mL) containing molecular sieves (3 Å), sodium 4-nitrophenolate (1.77 g, 11 mmol) was added. The reaction mixture was stirred at rt until TLC (1:1 EtOAc–hexane) showed the complete transformation of 4V. Nagy et al. / Carbohydrate Research xxx (2009) xxx–xxx CAR 4933 No. of Pages 7, Model 5G 20 February 2009 Disk Used ARTICLE IN PRESS Please cite this article in press as: Nagy, V. et al., Carbohydr. Res. (2009), doi:10.1016/j.carres.2009.02.011 UNCORRECTED PROOF the starting sugar (1 d). Then the mixture was filtered on a Celite pad and the solvent was removed. The oily residue was purified by column chromatography (1:1 EtOAc–hexane) to give 10 (0.93 g, 82%) as white crystals from EtOH. Mp: 233–235 °C; [ a ] D 320 +27 (c1.10, CHCl 3 ); 1 H NMR (CDCl 3 , 360 MHz): d(ppm) 8.13 (d, 2H, J9.2 Hz, ArH), 7.37 (d, 2H, J9.2 Hz, ArH), 6.87 (s, 1H, NH), 6.67 (s, 1H, NH), 5.77 (dd, 1H, J 2,3 9.8 Hz, J 3,4 2.6 Hz, H-3), 5.54 (d, 1H, J 2,3 9.8 Hz, H-2), 5.50 (dd, 1H, J 3,4 2.6 Hz, J 4,5 1.3 Hz, H-4), 5.00 (pseudo t, 1H, J 5,6 6.5 Hz, J 5,6 0 6.5 Hz, H-5), 4.13 (m, 2H, H-6, H-6 0 ), 2.11, 1.99 (2), 1.96 (3 s, 12H, OCOCH 3 ), 13 C NMR (CDCl 3 , 90 MHz): d(ppm) 170.1 (CONH 2 , 3 J H-2,CO =5.8 Hz), 169.5, 169.4, 168.8 168.3 (CO), 157.2, 143.9, 124.8, 120.9, (ArC), 99.8 (C-1), 72.4, 69.8, 67.0, 66.1 (C-2 to C-5), 61.2 (C-6), 20.4, 20.3 (3) (COCH 3 ). Anal. Calcd for C 21 H 24 N 2 O 13 (512.43): C, 49.22; H, 4.72; N, 5.47. 330 Found: C, 49.05; H, 4.66; N, 5.32. 1.14. C-[1-(4-nitrophenoxy)- a - D -galactopyranosyl]formamide ((4-nitrophenyl bD -galacto-hept-2-ulopyranoside)onamide) (11) To a solution of 10 (0.20 g, 0.39 mmol) in dry MeOH (5 mL) some crystals of KCN (5 mg) were added. The reaction mixture was stirred at rt until TLC (7:3CHCl 3 –MeOH)showed thecomplete transformation of the starting material (1 d). The reaction mixture was neutralised with a cation exchange resin Amberlyst 15 (H + form). After filtration, the solvent was removed to give 11 (0.15 g, 99%) as a yellowish oil. R f = 0.65 (7: 3 CHCl 3 –MeOH); [ a ] D +3 (c0.17, H 2 O); 340 1 H NMR (D 2 O, 360 MHz): d(ppm) 8.21 (d, 2H, J8.6 Hz, ArH), 7.45 (d, 2H, J8.6 Hz, ArH), 4.51 (pseudo t, 1H, J 5,6 6.8 Hz, J 5,6’ 5.1 Hz, H5), 4.15–4.10 (m, 3H, H-2, H-3, H-4), 3.86–3.78 (m, 2H, H-6, H-6 0 ); 13 C NMR (D 2 O, 90 MHz): d(ppm) 170.9 (CONH 2 , 3 J H-2,CO =4.6 Hz), 158.5, 143.5, 125.6 (2), 120.8 (2) (Ar), 101.8 (C-1), 76.9, 70.5, 69.5, 68.2 (C-2 to C-5), 61.4 (C-6). Anal. Calcd for C 13 H 16 N 2 O 9 (344.28): C, 45.35; H, 4.68; N, 8.14. Found: C, 45.33; H, 4.67; N, 8.10. 1.15. C-(2,3,4,6-Tetra-O-benzoyl-1-ethoxya - D -glucopyranosyl)- formamide ((ethyl 3,4,5,7-tetra-O-benzoyl-bD -gluco-hept-2ulopyranoside)onamide) (12) 350 This compound was prepared from 2(0.20 g, 0.28 mmol) according to General procedure I, and was purified by column chromatography (1:1 EtOAc–hexane) to give 12 (0.16 g, 87%) as a white crystalline product, and in the second fraction it gave compound 16 (0.02 g, 10%). Characterisation of 12: mp 88–91 °C; [ a ] D +65 (c 1.08, CHCl 3 ); 1 H NMR (CDCl 3 , 360 MHz): d(ppm) 8.01–7.24 (m, 20H, ArH), 6.82 (s, 1H, NH), 6.62 (t, 1H, J8.8 Hz, J8.8 Hz, H-3 or H4), 6.18 (s, 1H, NH), 5.88–5.80 (m, 2H, H-2, H-3 or H-4), 5.09 (ddd, 1H, J 4,5 8.8 Hz, J 5,6 6.7 Hz, J 5,6 0 3.2 Hz, H-5), 4.73 (dd, 1H, J 6,6 0 12.1 Hz, J 5,6 6.7 Hz, H-6), 4.41 (dd, 1H, J 6,6 0 12.1 Hz, J 5,6 0 3.2 Hz, H360 6 0 ), 3.89 (q, 2H, J6.8 Hz, CH 2 ), 1.18 (t, 3H, J6.8 Hz, CH 3 ); 13 C NMR (CDCl 3 , 90 MHz): d(ppm): 169.8 (CONH 2 , 3 J H-2,CO =4.1 Hz), 166.0, 165.3, 164.9 (2) (CO), 133.4–127.5 (ArC), 97.5 (C-1), 72.1, 72.0, 69.2, 68.8 (C-2 to C-5), 62.5 (C-6), 58.7 (CH 2 ), 15.2 (CH 3 ). Anal. Calcd for C 37 H 33 NO 11 (667.68): C, 66.56; H, 4.98; N, 2.10. Found: C, 65.75; H, 4.87; N, 2.22. 1.16. C-(1-Ethoxya - D -glucopyranosyl)formamide ((ethyl bD - gluco-hept-2-ulopyranoside)onamide) (13) This compound was prepared from 12 (0.06 g, 0.13 mmol) according to General procedure IV, and was purified by column 370 chromatography (7:2:1 CHCl 3 –MeOH–EtOAc) to give 13 (0.02 g, 98%) as a colourless oil. R f = 0.25 (7:2:1 CHCl 3 –MeOH–EtOAc); [ a ] D +18 (c0.37, H 2 O); 1 H NMR (D 2 O, 200 MHz): d(ppm) 3.90– 3.54 (m, 8H, H-2, H-3, H-4, H-5, H-6, H-6 0 ,CH 2 ), 1.22 (pseudo t, 3H, J7.0 Hz, J6.8 Hz, CH 3 ); 13 C NMR (D 2 O, 50 MHz): d(ppm) 172.6 (CONH 2 ), 99.7 (C-1), 76.4, 75.0, 73.1, 69.5 (C-2 to C-5), 61.4 (C-6), 59.7 (CH 2 ), 15.0 (CH 3 ). Anal. Calcd for C 9 H 17 NO 7 (251.24): C, 43.03; H, 6.82; N, 5.58. Found: C, 43.12; H, 6.75; N, 5.47. 1.17. C-(2,3,4,6-Tetra-O-benzoyl-1-n-buthoxya - D -glucopyranosyl)formamide ((n-buthyl 3,4,5,7-tetra-O-benzoyl-bD -gluco380 hept-2-ulopyranoside)onamide) (14) This compound was prepared from 2(0.50 g, 0.71 mmol) according to General procedure I, and was purified by column chromatography (1:2EtOAc–hexane)to give 14 (0.27 g, 56%) as a whitecrystalline product,andinthesecondfractionitgave compound16 (0.14 g,31%). Characterisation of 14:mp171–173°C; [ a ] D +64 (c1.10, CHCl 3 ); 1 H NMR (CDCl 3 ,360MHz):d(ppm) 8.10–7.24 (m, 20H, ArH), 6.79 (s, 1H,NH),6.63(t,1H,J9.2 Hz, J9.2 Hz, H-3 or H-4), 6.39 (s, 1H, NH), 5.88–5.82 (m, 2H, H-2, H-3 or H-4), 5.09 (ddd, 1H, J 4,5 9.2 Hz, J 5,6 3.5, J 5,6 0 3.0 Hz, H-5), 4.75 (dd, 1H, J 6,6 0 12.1 Hz, J 5,6 3.5 Hz, H-6), 4.40 (dd, 390 1H, J 6,6 0 12.1 Hz, J 5,6 0 3.0 Hz, H-6 0 ), 3.85–3.80 (m, 2H, CH 2 ), 1.57–1.50 (m, 2H, CH 2 ), 1.32–1.22 (m, 2H, CH 2 ), 0.83 (t, 3H, J7.1 Hz, J6.8 Hz, CH 3 ). 13 C NMR (CDCl 3 ,90MHz):d(ppm) 169.9 (CONH 2 , 3 J H2,CO =4.7 Hz), 165.3 (2), 164.9 (2) (CO), 133.4–128.1 (ArC), 97.5 (C1), 72.2, 72.1, 69.3, 68.9 (C-2 to C-5), 62.6 (C-6), 62.5, 31.6, 19.0 (CH 2 ), 13.6 (CH 3 ); Anal. Calcd for C 39 H 37 NO 11 (695.74): C, 67.33; H, 5.36; N, 2.01. Found: C, 66.95; H, 5.47; N, 2.32. 1.18. C-(2,3,4,6-Tetra-O-acetyl-1-deoxy-1-phenylaminoa - D - galactopyranosyl)formamide ((N-phenyl 3,4,5,7-tetra-Oacetyl-bD -galacto-hept-2-ulopyranosylamine)onamide) (17) 400 This compound was prepared from 1(0.20 g, 0.44 mmol) according to General procedure II. The oily residue was crystallised from Et 2 O to give 17 (0.16 g, 75%) as a white crystalline product. Mp: 200–201 °C, [ a ] D 30 (c1.02, CHCl 3 ); 1 H NMR (CDCl 3 , 360 MHz): d (ppm) 7.25–7.10 (m, 2H, ArH), 6.88–6.75 (m, H, ArH), 6.47 (s, 1H, NH), 6.02 (s, 1H, NH), 5.56 (dd, 1H, J 3,4 2.9 Hz, H-3), 5.52 (dd, 1H, J 4,5 0.9 Hz, H-4), 5.42 (pseudo t, 1H, J 5,6 7.2 Hz, H-5), 5.38 (d,1H, J 2,3 9.8 Hz, H-2), 5.05 (s, 1H, NH), 4.08 (dd, 1H, J 6,6 0 11.1 Hz, Hz, H-6), 4.02 (dd, 1H, J 5,6 0 6.8 Hz, H-6 0 ), 2.11 (2), 1.99, 1.96 (3 s, 12H, OCOCH 3 ); 13 C NMR (CDCl 3 , 90 MHz) d(ppm) 171.3, 170.9, 170.3, 410 170.1, 169.5 (CO), 141.8, 128.9 (2), 120.7, 117.2 (2) (ArC), 86.82 (C1), 71.4, 70.2, 68.7, 67.8 (C-2 to C-5), 61.9 (C-6), 20.8, 20.5 (3) (COCH 3 ). Anal. Calcd for C 21 H 26 N 2 O 10 (466.45): C, 54.08; H, 5.62; N, 6.01. Found: C, 54.88; H, 5.50; N, 5.84. 1.19. C-(2,3,4,6-Tetra-O-benzoyl-1-deoxy-1-phenylaminoa - D - glucopyranosyl)formamide ((N-phenyl 3,4,5,7-tetra-O-benzoylbD -gluco-hept-2-ulopyranosylamine)onamide) (18) This compound was prepared from 2(0.50 g, 0.71 mmol) according to General procedure II and was purified by column chromatography (1:2 EtOAc–hexane) to give 18 (0.26 g, 55%) from EtOH as 420 yellowish crystals. Mp 96–97 °C[ a ] D +108 (c1.33, CHCl 3 ); 1 HNMR (CDCl 3 , 360 MHz): d(ppm) 8.11–6.80 (m, 25H, ArH), 6.56 (s, 1H, NH), 6.36 (t, 1H, J9.1 Hz, J9.1 Hz, H-3 or H-4), 5.90–5.68 (m, 3H, H2, H-3 or H-4, NH), 5.27 (s, 1H, NH), 5.11–4.55 (m, 2H, H-5, H-6), 4.45 (dd,1H, J 6,6 0 12,1 Hz, J 5,6’ 3.2, Hz, H-6 0 ); 13 C NMR (CDCl 3 , 90 MHz): d(ppm) 171.2 (CONH 2 , 3 J H-2-CO =4.7 Hz), 166.2 (2), 165.5, 165.4 (CO), 142.0, 128.6 (2), 121.0, 117.5 (2) (ArC), 134.1– 128.3 (benzoyl ArC), 87.0 (C-1), 73.7, 72.8, 70.9, 69.7 (C-2 to C-5), 64.0 (C-6). Anal. Calcd for C 41 H 34 N 2 O 10 (714.74): C, 68.90; H, 4.72; N, 3.92. Found: C, 68.65; H, 4.80; N, 3.26. 430 1.20. C-(1-Deoxy-1-phenylaminoa - D -glucopyranosyl)formamide ((N-Phenyl bD -gluco-hept-2-ulopyranosylamine)onamide) (19) This compound was prepared from 18 (0.15 g, 0.21 mmol) according to General procedure IV, and was purified by column V. Nagy et al. / Carbohydrate Research xxx (2009) xxx–xxx 5 CAR 4933 No. of Pages 7, Model 5G 20 February 2009 Disk Used ARTICLE IN PRESS Please cite this article in press as: Nagy, V. et al., Carbohydr. Res. (2009), doi:10.1016/j.carres.2009.02.011 UNCORRECTED PROOF chromatography (7:2:1 CHCl 3 –MeOH–EtOAc) to give 19 (0.039 g, 54%) as a yellowish crystalline product. Mp 140–143 °C; [ a ] D +115 (c0.212, H 2 O); 1 H NMR (D 2 O, 360 MHz): d(ppm) 7.24–6.82 (m, 5H, ArH), 3.84–3.77 (m, 3H, H-5, H-6, H-6 0 ), 3.61 (d, 1H, J 2,3 9.2 Hz, H-2), 3.60–3.51 (m, 2H, H-3, H-4); 13 C NMR (D 2 O, 90 MHz): d(ppm) 175.4 (CONH 2 , 3 J H-2,CO =4.0 Hz), 144.5, 129.9 440 (2), 120.1, 115.6 (2) (ArC), 88.7 (C-1), 74.5, 74.4, 73.1, 69.7 (C-2 to C-5), 60.9 (C-6). Anal. Calcd for C 13 H 18 N 2 O 6 (298.30): C, 52.35; H, 6.08; N, 9.39. Found: C, 52.44; H, 6.23; N, 9.16. 1.21. C-(2,3,4,6-Tetra-O-benzoyl-1-deoxy-1-phenylsulfanyla - D - glucopyranosyl)formamide ((phenyl 3,4,5,7-tetra-O-benzoyl-2thio-bD -gluco-hept-2-ulopyranoside)onamide) (20) This compound was prepared from 2(0.50 g, 0.70 mmol) according to General procedure III, and was purified by column chromatography (1:2 EtOAc–hexane) to give 20 (0.41 g, 79%) as a white crystalline product. Mp: 89–92 °C; [ a ] D +33 (c0.25, CHCl 3 ); 450 1 H NMR (CDCl 3 , 360 MHz): d(ppm) 8.06–7.14 (m, 25H, ArH), 6.59 (s, 1H, NH 2 ), 6.51 (s, 1H, NH 2 ), 6.11, 5.78, (2 pseudo t, 2H, J9.2 Hz in each, H-3, H-4), 5.72 (d, 1H, J 2,3 9.2 Hz, H-2), 4.81– 4.76 (m, 2H, H-5, H-6), 4.46 (dd, 1H, J= 11.9, 4.0 Hz, H-6 0 ); 13 C NMR (CDCl 3 , 90 MHz) d(ppm): 168.1 (CONH 2 , 3 J H-2,CO =4.6 Hz), 166.0, 165.4, 164.9, 164.4 (CO), 136.6, 133.2 (2), 129.7 (3) (thiophenyl), 133.1–127.2 (ArCbenzoyl), 88.8 (C-1), 73.4, 71.9, 71.2, 68.8 (C-2 to C-5), 62.6 (C-6); Anal. Calcd for C 41 H 33 NO 10 S (731.28): C, 67.30; H, 4.55; N, 1.91. Found: C, 67.35; H, 4.59; N, 1.96. 460 1.22. C-(1-Deoxy-1-phenylsulfanyla - D -glucopyranosyl)formamide ((phenyl 2-thio-bD -gluco-hept-2-ulopyranoside)onamide) (21) This compound was prepared from 20 (0.20 g, 0.27 mmol) according to General procedure IV, and was purified by column chromatography (7:3 CHCl 3 –MeOH) to give 21 (0.07 g, 86%) as a colourless oil. R f = 0.74 (1:1 CHCl 3 –MeOH); [ a ] D +64 (c0.19, H 2 O); 1 H NMR (D 2 O 360 MHz): d(ppm) 7.72–7.46 (m, 5H, ArH), 3.92 (dd, 1H, J 6,6 0 13.2 Hz, J 5,6 0 1.0 Hz, H-6), 3.80 (dd, 1H, J 6,6 0 13.2 Hz, J 5,6 0 4.0 Hz, H-6 0 ), 3.64 (t, 1H, J9.2 Hz, J9.2 Hz, H-3 or H-4), 3.60–3.51 (m, 3H, H-2, H-3 or H-4, H-5); 13 C NMR (D 2 O 470 90 MHz): d(ppm) 172.2 (CONH 2 , 3 J H-2,CO =5.8 Hz), 137.3 (2), 130.9, 129.7 (2), 128.0 (tiophenyl), 89.1 (C-1), 78.2, 74.8, 74.6, 69.4 (C-2 to C-5), 61.0 (C-6), Anal. Calcd for C 13 H 17 NO 6 S(315.35): C, 49.52; H, 5.43; N, 4.44. Found: C, 49.57; H, 5.38; N, 4.48. 1.23. C-[2,3,4,6-Tetra-O-benzoyl-1-deoxy-1-(2-pyridylsulfanyl)- a - D -glucopyranosyl]formamide ((2-pyridyl 3,4,5,7-tetra-O-benzoyl-2-thio-bD -gluco-hept-2-ulopyranoside)onamide) (22) This compound was prepared from 2(0.70 g, 0.98 mmol) according to General procedure III, and was purified by column chromatography (1:1 EtOAc–hexane) to give 22 (0.56 g, 73%) as a 480 yellow crystalline product. Mp: 78–80 °C; [ a ] D +62 (c0.17, CHCl 3 ); 1 H NMR (CDCl 3 , 360 MHz): d(ppm) 8.33 (d, 1H, J2.6 Hz, pyridine), 8.06–7.23 (m, 23H, ArH, pyridine), 7.04 (s, 1H, NH 2 ), 6.60 (s, 1H, NH 2 ), 6.21 (pseudo t, 1H, J9.2 Hz, J9.2 Hz, H-3 or H-4), 6.05 (d, 1H, J 2,3 9.2 Hz, H-2), 5.89 (pseudo t, 1H, J10.6 Hz, J9.2 Hz, H-3 or H-4), 4.99 (ddd, 1H, J10.6 Hz, J4.0 Hz, J2.6 Hz, H-5), 4.73 (dd, H, J11.9 Hz, J2.6 Hz, H-6), 4.49 (dd, 1H, J 6,6 0 11.9 Hz, J 5,6 4.0 Hz, H-6 0 ); 13 C NMR (CDCl 3 , 90 MHz): d(ppm) 168.6 (CONH 2 , 3 J H-2,CO =5.9 Hz), 165.8, 165.3, 164.9, 164.4 (CO), 152.5, 149.4, 136.8, 133.1, 122.7 (pyridine), 133.4–128.1 (ArCbenzoyl), 88.1 490 (C-1), 73.7, 71.9, 71.6, 68.9 (C-2 to C-5), 62.8 (C-6); Anal. Calcd for C 40 H 32 N 2 O 10 S (732.77): C, 65.57; H, 4.40; N, 3.82. Found: C, 65.58; H, 4.36; N, 3.86. 1.24. C-[1-Deoxy-1-(2-pyridylsulfanyl)- a - D -glucopyranosyl]- formamide ((2-pyridyl 2-thio-bD -gluco-hept-2-ulopyranoside)- onamide) (23) This compound was prepared from 22 (0.20 g, 0.27 mmol) according to General procedure IV, and was purified by column chromatography (7:3 CHCl 3 –MeOH) to give 23 (0.05 g, 62%) as a colourless oil. R f = 0.64 (7:3 CHCl 3 –MeOH); [ a ] D +53 (c0.28, 500 H 2 O); 1 H NMR (D 2 O, 360 MHz): d(ppm) 8.59–7.55 (m, 4H, pyridine), 3.93 (dd, 1H, J 6,6 0 11.9 Hz, J 5,6 1.0 Hz, H-6), 3.84 (dd, 1H, J 6,6 0 11.9 Hz, J 5,6 0 2.6 Hz, H-6 0 ), 3.72 (t, 1H, J9.2 Hz, J9.2 Hz, H-3 or H4), 3.66–3.58 (m, 3H, H-2, H-3 or H-4, H-5); 13 C NMR (D 2 O, 90 MHz): d(ppm) 171.8 (CONH 2 , 3 J H-2,CO =5.8 Hz), 150.6, 150.4, 139.2, 133.1, 125.5 (pyridine), 89.3 (C-1), 78.2, 74.8 (2), 69.2 (C-2 to C-5), 60.8 (C-6); Anal. Calcd for C 12 H 16 NO 6 S(316.24): C, 45.56; H, 5.10; N, 8.86. Found: C, 45.59; H, 5.13; N, 8.89. 1.25. C-[2,3,4,6-Tetra-O-benzoyl-1-deoxy-1-(2-benzothiazolylsulfanyl)- a - D -glucopyranosyl]formamide ((2-Benzothiazolyl 510 3,4,5,7-tetra-O-benzoyl-2-thio-bD -gluco-hept-2-ulopyranoside)onamide) (24) This compound was prepared from 2(0.60 g, 0.84 mmol) according to General procedure III, and was purified by column chromatography (1:1 EtOAc–hexane) to give 24 (0.51 g, 76%) as a yellow crystalline product. Mp: 105–108 °C; [ a ] D 9(c0.17, CHCl 3 ); 1 H NMR (CDCl 3 , 360 MHz): d(ppm) 8.08–7.10 (m, 24H, ArH, benzothiazole), 7.24 (s, 1H, NH 2 ), 6.37 (s, 1H, NH 2 ), 6.18 (t, 1H, J9.2 Hz, J9.2 Hz, H-3 or H-4), 6.07 (d, 1H, J 2,3 9.2 Hz, H-2), 5.98 (t, 1H, J9.2 Hz, J9.2 Hz, H-3 or H-4), 5.06 (ddd, 1H, J 4,5 520 9.2 Hz, J 5,6 4.0 Hz, J 5,6 0 1.0 Hz, H-5), 4.84 (dd, 1H, J 6,6 0 13.2 Hz, J 5,6 4.0 Hz, H-6), 4.57 (dd, 1H, J 6,6 0 13.2 Hz, J 5,6 0 1.0 Hz, H-6 0 ); 13 CNMR (CDCl 3 , 90 MHz): d(ppm) 167.4 (CONH 2 , 3 J H-2,CO =4.9 Hz), 165.9, 165.3, 164.9, 164.3 (CO), 157.1, 152.2, 137.3, 126.2, 125.2, 123.0, 120.9 (benzothiazole), 133.7–128.2 (ArCbenzoyl), 88.7 (C1), 74.3, 71.5 (2), 68.6 (C-2 to C-5), 62.8 (C-6); Anal. Calcd for C 42 H 32 N 2 O 10 S 2 (788.86): C, 63.95; H, 4.09; N, 3.55. Found: C, 63.99; H, 4.11; N, 3.50. 1.26. C-[1-Deoxy-1-(2-benzothiazolylsulfanyl)- a - D -glucopyranosyl]formamide ((2-benzothiazolyl 2-thio-bD -gluco-hept-2530 ulopyranoside)onamide) (25) This compound was prepared from 24 (0.20 g, 0.25 mmol) according to General procedure IV, and was purified by column chromatography (7:3 CHCl 3 –MeOH) to give 25 (0.04 g, 47%) as a yellow crystalline product. Mp: 183–185 °C; [ a ] D +95 (c0.27, DMSO); 1 H NMR (DMSO-d 6 , 360 MHz): d(ppm) 8.07–7.41 (m, 4H, benzothiazole), 7.78 (s, 1H, NH 2 ), 7.54 (s, 1H, NH 2 ), 6.26 (d, 1H, J5.3 Hz, OH), 5.32 (d, 1H, J4.0 Hz, OH), 5.11 (d, 1H, J5.3 Hz, OH), 4.56 (pseudo t, 1H, J5.3 Hz, J4.0 Hz, OH), 3.78 (dd, 1H, J 6,6 0 11.9 Hz, J 5,6 6.6 Hz, H-6), 3.68–3.56 (m, 5H, H-2, H-3, H-4, H-5, 540 H-6 0 ); 13 C NMR (DMSO-d 6 , 90 MHz): d(ppm) 169.6 (CONH 2 , 3 J H-2,CO =5.9 Hz), 159.9, 151.7, 136.9, 126.1, 125.2, 122.1, 121.4 (benzothiazole), 87.8 (C-1), 79.1, 74.7, 74.5, 69.1 (C-2 to C-5), 60.9 (C-6), Anal. Calcd for C 14 H 16 NO 6 S 2 (372.42): C, 45.15; H, 4.33; N, 7.52. Found: C, 45.05; H, 4.35; N, 7.54. 1.27. X-ray data collection and reduction Crystals of 4were grown from EtOAc by slow evaporation of the solution. A colourless block crystal (0.67 0.56 0.4 mm) was 6V. Nagy et al. / Carbohydrate Research xxx (2009) xxx–xxx CAR 4933 No. of Pages 7, Model 5G 20 February 2009 Disk Used ARTICLE IN PRESS Please cite this article in press as: Nagy, V. et al., Carbohydr. Res. (2009), doi:10.1016/j.carres.2009.02.011 UNCORRECTED PROOF fixed on a glass capillary using epoxy glue. Data were collected at 293(1) K, Bruker-Nonius MACH3 diffractometer, Mo K a radiation 550 k= 0.71073 Å, x motion, h max =25.4°.The structure was solved using the SIR-92 software 23 and was refined on F 2 using SHELX -97 program, 24 publication material was prepared with the WINGX - suite. 25 Crystal data: formula C 17 H 25 NO 11 ,M= 419.38, monoclinic, space group P2 1 ,a= 8.569(2) Å, b= 18.397(6) Å, c= 13.688(8), b=94.32(2)°,V= 2174(2) Å 3 ,Z=4, q calcd = 1.281, 4540 measured, 2899 reflections were unique with I>2 r (I), decay: 3%, R 1 = 0.088 and wR 2 = 0.241 for 4074 reflections and 503 parameters, GOF = 1.11. Residual electron density: 0.7/0.31 e/Å 3 . Hydrogen atoms were fixed into geometric position except N–H 560 hydrogens which could be found at the difference electron density map, but were also fixed into calculated positions in the final stage of the refinement. There is a remaining electron density (0.7 e  /Å 3 ) close to the acetyl carbon atom of C16 which may indicate some disorder of this acetyl group. However, this has no effect on our main findings concerning the configuration of the anomeric carbon. Anisotropic refinement of non-hydrogen atoms was performed except atoms of the C16 acetyl group. Orientation of methyl groups was refined using a riding model. There are two molecules found in the asymmetric unit with slightly different bond length and angle 570 data as indicated in Figure 1, too. The structure is stabilised with intermolecularhydrogenbondsbetweentheamidehydrogenatoms and the O8 acetylene or O11 amide carbonyl oxygen atoms ofa symmetry related molecule. Intramolecular hydrogen bond between O1 and the amide proton causes nearly planar orientation of O1–C1– C7–O11–N1. The uniqueness of compound 4is shown by the fact that no similar structure could be found in the Cambridge Structural Database 26 (Ver. 5.29, November 2007 with upgrades in 2008) containing an amide group as well as oxygenconnected to the anomeric carbon atom. Additional crystallographic information is provided in 580 the deposited CIF: CCDC 714419. Acknowledgements This work was supported by the Hungarian Scientific Research Fund (Grants: OTKA 46081 and 61336). The authors thank P. Gergely and T. Docsa for the glycogen phosphorylase assays. References 1. Kiss, L.; Somsák, L. Carbohydr. Res. 1996,291, 43–52. 2. } Osz,E.;Sós, E.;Somsák,L.;Szilágyi,L.;Dinya,Z. Tetrahedron 1997,53,5813–5824. 3. } Osz, E.; Somsák, L.; Szilágyi, L.; Kovács, L.; Docsa, T.; Tóth, B.; Gergely, P. Bioorg. Med. Chem. Lett. 1999,9, 1385–1390. 5904. Somsák, L.; Nagy, V.; Docsa, T.; Tóth, B.; Gergely, P. Tetrahedron: Asymmetry 2000,11, 405–408. 5. Somsák, L.; Nagy, V. Tetrahedron: Asymmetry 2000,11, 1719–1727. Corrigendum 2247. 6. Somsák, L.; Kovács, L.; Tóth, M.; } Osz, E.; Szilágyi, L.; Györgydeák, Z.; Dinya, Z.; Docsa, T.; Tóth, B.; Gergely, P. J. Med. Chem. 2001,44, 2843–2848. 7. Somsák, L.; Sós, E.; Györgydeák, Z.; Praly, J.-P.; Descotes, G. Tetrahedron 1996, 52, 9121–9136. 8. Czifrák, K.; Kovács, L.; Kövér, K. E.; Somsák, L. Carbohydr. Res. 2005,340, 2328– 2334. 6009. Gyóllai, V.; Somsák, L.; Szilágyi, L. Tetrahedron Lett. 1999,40, 3969–3972. 10. Somsák, L.; Kovács, L.; Gyóllai, V.; } Osz, E. Chem. Commun. 1999, 591–592. 11. Oikonomakos, N. G. Curr. Protein Pept. Sci. 2002,3, 561–586. 12. Somsák, L.; Nagy, V.; Hadady, Z.; Docsa, T.; Gergely, P. Curr. Pharm. Des. 2003,9, 1177–1189. 13. Somsák, L.; Nagy, V.; Hadady, Z.; Felföldi, N.; Docsa, T.; Gergely, P. In Frontiers in Medicinal Chemistry; Reitz, A. B.; Kordik, C. P.; Choudhary, M. I.; Rahman, A. u. Eds.; Bentham, 2005; pp 253–272. 14. Oikonomakos, N. G.; Somsák, L. Curr. Opin. Invest. Drugs 2008,9, 379–395. 15. Somsák, L.; Czifrák, K.; Tóth, M.; Bokor, É.; Chrysina, E. D.; Alexacou, K. M.; 610Hayes, J. M.; Tiraidis, C.; Lazoura, E.; Leonidas, D. D.; Zographos, S. E.; Oikonomakos, N. G. Curr. Med. Chem. 2008,15, 2933–2983. 16. Watson, K. A.; Mitchell, E. P.; Johnson, L. N.; Son, J. C.; Bichard, C. J. F.; Orchard, M. G.; Fleet, G. W. J.; Oikonomakos, N. G.; Leonidas, D. D.; Kontou, M.; Papageorgiou, A. Biochemistry 1994,33, 5745–5758. 17. Krülle, T. M.; Fuente, C.; Watson, K. A.; Gregoriou, M.; Johnson, L. N.; Tsitsanou, K. E.; Zographos, S. E.; Oikonomakos, N. G.; Fleet, G. W. J. Synlett 1997, 211–213. 18. Chrysina, E. D.; Oikonomakos, N. G.; Zographos, S. E.; Kosmopoulou, M. N.; Bischler, N.; Leonidas, D. D.; Kovács, L.; Docsa, T.; Gergely, P.; Somsák, L. 620Biocatal. Biotransform. 2003,21, 233–242. 19. Fügedi, P. In The Organic Chemistry of Sugars; Levy, D. E., Fügedi, P., Eds.; CRC Press: Boca Raton, 2006; pp 89–179. 20. Eellis, G. P.; Honeyman, J. Adv. Carbohydr. Chem. 1955,10, 95–168. 21. Paulsen, H.; Pflughaupt, K. W. In The Carbohydrates, Chemistry and Biochemistry; Pigman, W., Horton, D., Eds.; Academic Press: New York, 1980; pp 881–927. 22. El Ashry, E. S. H.; Awad, L. F.; Atta, A. I. Tetrahedron 2006,62, 2943–2998. 23. Altomare, A.; Cascarano, G.; Giacovazzo, C.; Guagliardi, A. J. Appl. Crystallogr. 1993,26, 343–350. 24. Sheldrick, G. M. SHELXL -97 1997, (Programs for Crystal Structure Analysis, 630Release 97-2). Institut für Anorganische Chemie der Universität, Tammanstrasse 4, D-3400 Göttingen, Germany. 25. Farrugia, L. J. J. Appl. Crystallogr. 1999,32, 837–838. 26. Allen, F. H. Acta Crystallogr. 2002,B58, 380–388. doi:10.1107/ S0108768102003890. V. Nagy et al. / Carbohydrate Research xxx (2009) xxx–xxx 7 CAR 4933 No. of Pages 7, Model 5G 20 February 2009 Disk Used ARTICLE IN PRESS Please cite this article in press as: Nagy, V. et al., Carbohydr. Res. (2009), doi:10.1016/j.carres.2009.02.011