MAGNETIC RESONANCE IN CHEMISTRY, VOL. 29, 852-858 (1991)

Reference Data

'H and Chemical Shift Assignments of para-Substituted Aryl 2-Acetamido-2- deox y-/?-D-glucop yr anosides

RENe ROY, FRANCOIS D. TROPPER and ANTONY J. WILLIAMS Department of Chemistry, University of Ottawa, Ottawa, Ontario, KIN 6N5, Canada

The 'H and '3C N M R spectra of twenty aryl 2-acetarnido-2-deoxy-/3-o-glucopyra- nosides and eighteen aryl 3,4,6-tri-U- acetyl- 2- acetamido- 2- deoxy-8- D -gluco - pyranosides have been obtained and assigned. The three-bond proton coupling constants of these compounds were also obtained.

KEYWORDS 13C N M R ' H N M R 'J(CH) Glucopyranosides Aryl 2-acetamido-2- deoxy-/3- 0-glucopyranosides

INTRODUCTION

Aryl glycosides are widely distributed in nature and many of these compounds have been shown to have medicinal properties and applications in enzymology' and lectinol- ogy.' A series of aryl 2-acetamido-2-deoxy-p- D-ghcopyranosides has been synthesized to probe the electronic contributions of various substituents to the relative binding associ- ations with the wheat germ lectin Triticurn uutgaris, which has tyrosyl residues in the carbohydrate binding sites. This paper reports the 'H and ',C chemical shifts and proton coupling constants derived from a number of acetylated and non-acetylated aryl 2-acetamido-2-deoxy-~-~-glucopyranosides.

EXPERIMENTAL

All compounds examined were prepared as discussed elsewhere.,- The 'H and I3C NMR spectra were obtained on a Varian Gemini 200 NMR spectrometer, except for the 'H-'H COSY and 'H-13C correlation

2D spectra, which were obtained on a Varian XL-300 spectrometer. Spectra were generally obtained in CDCI, for the acetylated com- pounds and in DMSO-d, for the non- acetylated compounds. In order to simplify the spectra, for most of the non-acetylated glycosides labile hydrogens were exchanged for deuterium by lyophilizing D,O solutions of these compounds. Occasionally, solubility problems necessitated the use of solvent mix- tures of 10% DMSO-CDCI,, or even the spectra being obtained in D,O with internal acetone as a reference standard (details are given in the tables). 'H spectra were obtained at 199.975 MHz and were referenced to the respective residual solvent peak (7.24 ppm for chloroform, 2.50 ppm for DMSO and 2.216 ppm for acetone in D,O). The I3C spectra were obtained at 50.283 MHz and again were referenced to the solvent or internal reference peak (CDCI, at 77.00 ppm, DMSO-d, at 39.49 ppm and acetone in D,O at 31.06 ppm). All spectra were obtained at ambient temperature (23 f 1 "C).

For the 'H spectra the spectral width was 3000 Hz defined by 17984 points. A 2.997 s acquisition time was used with a 4 ps (37') pulse and no delay between scans. The number of scans obtained was never less than 16 and was optimized to give a good signal- to-noise ratio in the spectra. Spectra were analysed using a first-order approximation.

For the I3C spectra the spectral width was 12 579 Hz defined by 25 088 points. A 0.997 s acquisition time was used with a 5 ps (39") pulse and no delay between scans. The number of scans obtained was never less than 320 and was optimized to give a good signal- to-noise ratio in the spectra. A 1.5 Hz expo- nential line broadening was applied to each of the spectra.

The homonuclear 'H-'H COSY experi- ment was obtained using the double- quantum filtered COSY pulse sequence at 299.94 MHz. The spectra were collected as 1024 x 512 blocks of data, and were pro- cessed using sinusoidal multiplication in each dimension. The raw data were zero-filled to 1024 words in the second dimension before double Fourier transformation. The final data matrix was symmetrized.

The heteronuclear 'H-l3C correlation experiment was obtained using the standard

sequence at a frequency of 75.43 MHz. The spectra were collected as 1024 x 512 blocks of data, and processed using sinusoidal multi- plication in each dimension. Resonance multiplicities for 13C were obtained using the DEPT technique with 'H pulses of 45", 90" and 135". The 'H 90" pulse was 45 ps, the 13C 90" pulse was 9.6 ps and the jJ(C1-I) delay was set to 3.6 ms.

RESULTS AND DISCUSSION

Figure 1 illustrates the 'H-'H COSY spec- trum obtained for p-chlorophenyl 3,4,6-tri-t3- acetyl-2-acetamido-2-deoxy-~-~-gluco-pyr- anoside. The full assignment of the 'H spec- trum was obtained by tracing the conned- vities starting with the known typical shift6 of the anomeric proton H-1. 'The assignment was confirmed with the 'H-I3C hetero- nuclear correlation spectrum.

Tables 1-6 give the 'H and 13C chemical shifts and 'J(HH) coupling constants for all compounds examined using the assignme:nt obtained for p-chlorophenyl 3,4,6-tri-0- acetylated - 2 - acetamido - 2 -deoxy -P-D -gluco - pyranoside.

- References

1. J. A. Cabezas, A. Reglero and P. Calvo, 1nr.J. Biochem. 15, 243 (1983).

2. I . E. Liener, N. Sharon and I. J. Golcl- stein, The Lectins-Properties, Func- tions and Applications in Biology and Medicine. Academic Press, Orlando (1 986).

3. R. Roy and F. D. Tropper, Synrh. Commun. 20,2097 (1 990).

4. R. Roy and F. D. Tropper, Can. J. Chem. 69, (1991) 817, and references cited therein.

5. R. Roy, F. D. Tropper, A. Rornanowska. M. Letellier, L. Cousineau, S. Meunier and J. Boratynski, Glycoconj. J. (1, (1 991 ) in press.

6. K. Bock and H. Thogersen, Annu. Rep. NMR Spectrosc. edited by G. A. Webb, Academic Press, New York, 13, 1 (1 983).

Received 25 October 1990; accepted (revised) 6 March 1991

Q 1991 by John Wiley & Sons, Ltd.

853

5 . 0 -

5 . 5 -

6.0-

6 . 5 -

Reference Data

ortho meta Y = Cl R = Ac

Acetates H m Ho H1

H3 H4

NH

F 2 (PPM 1

(d

i Q O

.' 0 m

0

a 0 9

U

7 . 0 6 . 5 6 . 0 5.5 5 . 0 4 . 5 4 . 0 3.5 3 . 0 2.5 2.0 1.5 F I (PPM)

Figure 1. ' H-' H COSY spectrum of p-chlorophenyl 3,4,6-tri-0-acetyl-2-acetamido-2-deoxy-~-o-glucop~ranoside. See text for experi- experimental conditions.

854

Reference Data

~ ~

Table 1. 'H NMR chemical shifts (ppm) of non-acetylated glycosides

Compound' X

H

CH,

t-Bu

Ph

F

CI

Br

I

OCH,

NHA,

NHCOCHCH,

NH,

NO2

Me,N+I-

CN

CHO COOCH,

COOH"

CH,OH

m-CH,

H - l (d)

4.95

4.86

4.89

4.99

4.85

4.91

4.92

4.91

4.92 4.68 4.47

4.1 0

4.91

5.30

5.07

5.14 5.09

5.22

5.1 6

4.90

H-2-H-6

3.67-3.77 (2H)

3.62-3.71 (2H)

3.63-3.73 (2H) 3.1 8-3.52 (4H) 3.67-3.78 (2H)

3.17-3.54 4(H)

3.12-3.51 (4H)

3.17-3.55 (4H)

3.66-3.75 (2H) 3.1 6-3.54 (4H) 3.66-3.75 (2H) 3.19-3.53 (4H) 3.63-3.77 (2H) 3.17-3.51 (4H) 3.65-3.74 (2H) 3.13-3.51 (4H)

3.14-3.70 (6H) 3.23-3.74 (6H)

3.13-3.75 (6H)

3.1 5-3.75 (6H)

3.65-3.74 (2H) 3.1 4-3.52 (4H) 3.944.07 (2H) 3.52-3.84 (4H) 3.67-3.78 (2H) 3.1 5-3.52 (4H) 3.20-3.74 (6H) 3.63-3.76 (2H) 3.1 7-3.55 (4H) 3.93-4.06 (2H)

3.91-4.02 (2H) 3.52-3.84 (4H)

3.50-3.82 (4H) 3.64-3.73 (2H) 3.1 8-3.52 (4H)

NH

b

b

b

b

b

b

b

b

b

b

7.81 d

7.81 d

b

b

b

b

7.82 d

b

b

b

H-meta (d) H-ortho (d) X

7.26 m 7.02-6.90 m

7.06 6.94 2.18 s

7.26 6.85 1.18 s

7.54 m (4H) 7.41 m (2H) 7.30 m (1 H) 7.13 d (2H)

6.91-7.10 m (4H) -

7.28 6.95 -

7.42 6.91 -

7.57 6.78 -

6.85 m (4H) 3.63 s 6.69 6.52 b

7.46 6.89 9.85 s (NH) 2.00 s (CH,)

trans 6.22 dd cis 5.72 dd

7.57 6.93 -CH- 6.91 dd

7.57 6.78 d -

7.80 7.25 3.64 s

7.72 7.09 -

7.87 7.14 9.89 s 7.90 7.05 3.81 s

7.78 7.08

7.36 7.07 4.58 s

2.21 s

b

7.1 3 m (1 H) 6.76 m (3H)

NAc ( 5 )

1.82

1.81

1.81

1.83

1.83

1.82

1 801

1.801

1.84 1.82 1.81

1.81

1.81

2.03,

1.80

1.791 1.791

2.02

2.02

1.81

"para-Substituents unless stated otherwise Exchanged protons, ND. Spectrum obtained in D,O with acetone as internal standard.

855

Reference Data

Table 2. "C NMR chemical shifts (ppm) of non-acetylated glycosides

Cornpound* X

H

I-Eu

Ph

CH,

F

CI 0r I OCH,

NHAc

NHCOCHCH,

NH2

NO2 Me,N+I- CN CHO COOCH,

COOH" CH,OH m-CH,

c -1

99 37 99 73 99 55

99 39

99 89

99 44 99 32 9916 99 52

101 12 99 82

99 73

98 56 99 78 98 52 98 71 98 29

10004 99 35 99 39

c -2

55.55 55.59 55.57

55.56

55.46

55.46 55.03 55.41 55.63 55.69 55.53

55.44

55.27 56.07 55.31 55.61 55.26

56.33 55.48 55.58

c - 3

74.15 74.13 74 14

74.10

74.02

74.03 74.00 73.98 74.16 74.19 74.13

74.04

73.89 74.10 73.95 74.21 73.86

74.43 73.95 74.16

c-4

70 41 70 43 70 45

69 39

70 29

70 31 70 28 70 26 70 48 70 54 70 42

70 26

7010 70 38 7018 70 50 70 05

70 57 70 25 70 42

c -5

77.31 77.26 77.28

77.35

77.24

77 32 77.31 77.29 77.26 77.14 77.25

77.29

77.45 76.89 77.41 77.42 77.23

77.06 77.1 3 77.31

C-6

60 80 60 82 60 83

60 80

60 72

60 73 60 70 60 68 60 88 60 93 60 79

60 70

60 55 61 20 60 62 60 94 60 50

61 38 62 41 60 80

IPS0

157 86 155 80 155 66

15743

15380d

156 55 156 98 157 59 15478 14922 153 50

153 90

t2Wb

162 52 15815 16089 162 54 16085

16012 156 26 157 90

mho

11656 11658 11609

11699

11580d (23 1 ) b

11836 11883 11923 11466 11465 11679

11694

11673 11859 11723 11694 11592

11684 11599 11349

11732 (C-2')

(C 6')

mera

129 65 129 91 126 18

1343

11795d

12941 13231 13817 11804 11816 12045

12073

(8 2)b

125 90 12246 134 37 131 05 13095

132 07 127 57 13917

122 90 (C-3')

(C-5)

para X

122 20 -

131 03 20 06 144 52 C 33 78

CH, 31 23 13991 ips0 134 30

onho 126 45 mera 127 90 para 127 06

15736d - (237 4)b 12597 - 11367 -

85 00 -

161 91 55 43 14402 -

134 04 23 70

126 83 C-0 163 04 -CH- 132 09

CH,- 133 67 14202 - 142 14 57 98 11903 104 44 13218 192 22 12298 165 52

51 86 131 67 175 73 135 98 70 67 129 38 21 00

168 1

NHCOCH,

169 60 169 54 169 53

16960

169 29

16958 16952 169 48 16961 16947 169 52

169 57

16964 175 74 16966 17055 169 05

17604 168 95 169 58

23 02 23 00 22 97

22 99

23 05

22 96 22 95 22 94 23 47 23 02 22 98

23 08

22 94 22 92 22 95 23 16 23 05

22 90 23 09 23 04

.para Substituents u n l w stated otherwise

CSpectrurn obtained in D,O with acetone as internal standard Coupling constants for 13C-'sF couplings ( H r )

856

Reference Data

Table 3. 'H NMR chemical shifts (ppm) of acetylated glycosides

Compound* X

H

CH,

t-Bu

Ph

F

CI

Br

I

OCH,

NH,

NHAc

NO,

CN

CHO

COOCH,

CH,OH

5.18 4.07 5.37 5.13 3.68 4.27 4.12 5.58 7.05 6.86

5.21 4.09 5.38 5.12 3.83 4.27 4.13 5.54 7.27 6.90

5.30 4.14 5.41 5.13 3.88 4.29 4.15 5.77 7.48 7.04

5.18 4.05 5.38 5.11 3.82 4.27 4.13 5.65 6.94 (2H)b

5.23 4.06 5.39 5.10 3.84 4.26 4.12 5.77 7.20 6.90

5.23 4.06 5.39 5.10 3.84 4.25 4.12 5.77 7.35 6.85

5.23 4.06 5.39 5.10 3.84 4.26 4.12 5.66 7.54 6.74

5.13 4.05 5.37 5.11 3.79 4.27 4.12 5.59 6.92 6.78

5.03 4.05 5.34 5.09 3.75 4.25 4.11 5.81 6.79 6.56

5.05 4.05 5.22 4.96 3.71 4.15 3.99 7.21 7.32 d 6.81 d

5.55 ND" 5.23 4.96 ND" ND" ND" 8.13 8.23 7.24

5.43 4.05 5.44 5.11 3.92 4.26 4.12 5.92 7.55 7.03

5.42 4.11 5.43 5.12 3.92 4.27 4.14 5.75 7.81 7.07

5.42 4.10 5.45 5.09 3.93 4.24 4.12 6.41 7.89 6.95

5.24 4.08 5.39 5.12 3.84 4.27 4.13 5.56 7.27 6.96

NHCOCHCH, 5.16 4.12 5.35 5.10 3.81 4.26 4.11 5.97 7.47 6.92

X

Combined rn centred at 7.01

2.27 s

1.26 s

7.40 broad m -

3.74 s

NH, 3.52 s

8.75 s, NH 1.78 Ac

(broad)

-

9.89 s

3.83 s

4.62 dd

COCH- 6.23 dd H-trans 5.73 dd

H-ck 6.41 dd

Ac (s)

1.94, 2.03 2.04, 2.06 1.93, 2.02 2.04, 2.06 1.93, 2.02 2.04, 2.0!5 1.95, 2.03 2.04, 2.06 1.94, 2.0;! 2.04, 2.05 1.92, 2.02 2.03, 2.04 1.92, 2.02 2.03, 2.04 1.92, 2.02 2.04, 2.05 1.95, 2.0'1 2.03, 2.063 1.93, 2.00 2.02, 2.04

1 .go, 1.90 1.94, 1.99 1.77, 1.9!5 2 x 2.00 1 .go, 2.02 2.03. 2.04 1.93, 2.03 2.04, 2.0!5 1.85, 2.00 2.01, 2.02 1.94, 2.03 2.04, 2.063 1.94, 2.02 2.03, 2.0!j

1.93. 2.02 6.77. 6.84 2.04, 2.05

m-CH, 5.22 4.06 5.38 5.11 3.85 4.26 4.12 5.57 7.14 6.79 s (H-2') 2.29 s

(H-4',6)

'para Substituents unless stated otherwise. bAppears as AB system: one doublet, J = 6.3 Hz.

Overlapping signals. Appears as AB system: one doublet, J = 3.6 Hz.

857

-

Reference Data

Table 4. I3C NMR chemical shifts (ppm) of acetylated glycosides

Compound'X C-1 C-2 C-3 C-4 C-5 C-6 ips0 ortho mefa para

H 98.92 54.66 71.83 68.56 72.03 62.10 157.17 116.96 129.60 123.20

CH3 9928 54.50 71.61 68.81 72.16 62.16 155.12 11702 129.87 132.50

f-Bu 98.88 5442 71 51 68.69 72.09 62.08 15483 116 39 126.22 14582

Phb 98.13 53.17 70.99 68.36 72 35 61.65 156.58 116.78 139.84. 13490. 2 x 128 83, 2 x 127.84

126.98, 2 x 126.44 F 99.67 54.65 71.83 68.70 72.00 62.11 161.21 115.85d 118.69d

(13 4)' CI 9879 5388 7147 6848 7198 6188 15562 11825

Br 9870 5449 7174 6857 7183 6203 15616 11879

I 9854 5349 71 24 6836 71 98 61 72 15683 11908

OCH, 10007 5449 71 68 6859 7206 6207 15566 11447

NHZd 10016 5665 7106 6853 7214 6178 14958 11528

NHAcd 9919 5331 7105 6833 7206 6163 15298 11694

NO, 9706 5296 71 16 6819 7225 6150 16171 11683

CN 9754 5343 71 47 6821 71 71 61 58 15988 11698

CHO 9772 5386 71 72 6850 71 95 61 88 161 69 11667

COOCH, 9779 5448 7180 6848 7187 6200 16057 11608

CH,OH 9905 5435 71 67 6859 7203 6209 15652 11693

NHCOCHCHZb 98 50 5339 71 08 6861 72 69 61 85 15321 117 21

m-CH, 9887 5468 7182 6869 7211 6222 15720 11375

11773 (C-2')

(C-6)

*para Substituents unless stated othemtse 'Spectrum obtained in DMSO-d, CCoupling constants are for '3C-1sF couplings dSpectrum obtained in 10% DMSO-d. in CDCI, solution

( 8 1 Y 12914

132 37

137 93

11863

11845

12084

12603

133 48

131 52

131 50

128 37

121 09

139 7 (C-3) 123 92 (C-5)

15484 d (156 5)'

127 69

11551

147 37

151 58

14241

133 80

14248

147 6

131 31

12465

135 90

134 32

12927

X C-O(Ac) CH,(NHAc) CH,(OAc)

-

20.41

c 33 95 CH, 31.21

-

-

-

-

55.03

-

23 34

-

105 57

19077

166 66 51 92 64 30

132 14 (-CH-) 12721 (CH,-) 16348 (C-0)

CH3 21 25

169.61, 171.02 170.64, 170.83 169 54, 170.70 2 x 170.80 169.58. 2 x 170.79 170 87 169.29. 169.72 169.81, 170.00 169.55 169.55. 170 69 170 79,170 85 169 40, 170.48 2x17079 169.55, 170.70 170.83. 170.92 16917.17012 170 19, 170.39 169 60, 170.71 170 83, 170 97 16920,17009 170.25, 170.34 168 51, 169.09 16997,17011 170 30 169.65. 169.95 170 03. 170 35

16909. 17012 170.45. 170.61 169.23. 2 x 171.41 17073 169.57. 17072 2x17087 169.70. 170.91 2 x 170.99 16993. 2 x 170.29 17061

169 58, 170.60 170.77. 170 97

2313

22 89

22 95

22 53

22 98

22 69

22 97

22 51

23 08

22 47

22 32

22 62

22 43

22 67

23 00

22 96

22 79

23 08

20 47 2 x 2 0 5 5 2 x 2 0 3 2

20 41 20 38

2 x 2 0 4 7 20 21 20 25 20 33 20 37

2 x 2 0 4 4 20 22

2 x 20 30 20 39

2 x 2 0 4 9 20 05

2 x 2 0 1 1 20 43

2 x 20 53 3 x 2 0 0 6

3 x 1 9 9 3

20 31 20 41 20 47

3 x 2 0 0 1

2018 2 x 2 0 2 4

20 41 2 x 20 51

20 45 2 x 2 0 5 2

20 55

20 42 20 47

2 x 2 0 5 0

858

Reference Data

Table 5. ‘H thee-bond coupling constants acetylated glycosides’

Cornpobnd”.b X

H

r-Bu Ph F CI Br I OCH,

NHAc NHCOCHCH,

CH 3

NH,

NO,

CN CHO COOCH, COOH CH,OH m-CH,

J(12 )

8.4 8.4 8.5 8.4 8.4 8.4 8.5 8.4 8.4 8.3 8.4 8.4

8.1 8.4 8.4 8.4 8.5 8.5 8.4 8.5

J(H-2. NH) J(om)

d

c

c 8.7 8.9

c d

c d

9.0 9.0 8.9

8.8 8.9 10.0 9.2 9.0

c

d

c

c

c 8.9 9.4 8.9 8.8

9.0 8.9 8.9 8.7 c

c d

(Hz) of non-

J(W d

- - d

- -

-

- - c

- J,,,,, = 17.1

J,,, = 9.8 J,,, = 2.6 -

- - - - - - -

“The proton signals from H-2 to H-6,6‘ give either two multiplets or a single multiplet as indicated in Table 1. bpara Substituents unless stated otherwise.

Exchanged. Unresolved.

Table 6. ‘H three-bond coupling constants (Hz) of acetylated glycosides

Compound” X J ( 1 2 ) J (23 ) J (34 ) J (45 ) J (56 ) J(56‘) J(66’)

H 8.0 10.2 9.4 9.5 5.2 2.6 12.3 CH, 8.2 10.5 9.3 9.3 5.4 2.5 12.2 t - B u 8.1 10.4 9.3 9.4 5.2 2.5 12.3 Ph 8.3 10.5 9.3 9.5 5.3 2.5 12.2 F 8.3 10.5 9.3 9.8 5.3 2.6 12.2 CI 8.2 10.5 9.3 9.5 5.4 2.5 12.2 Br 8.1 10.5 9.4 9.5 5.4 2.4 12.2 I 8.3 10.4 9.3 9.5 5.4 2.4 12.2 OCH, 7.5 10.0 9.3 9.6 5.2 2.5 12.3 NH, 8.4 10.5 9.3 9.6 5.2 2.5 12.2 NHA, 8.4 10.3 9.6 9.7 5.3 2.5 12.3 NO, 8.5 10.3 9.4 9.6 NHCOCHCH, 8.1 10.3 9.5 9.3 5.7 2.4 11.4 CN 8.1 10.2 9.6 9.5 5.6 2.3 12.3 CHO 8.1 10.4 9.2 9.4 5.5 2.5 12.2 COOCH, 8.4 10.5 9.5 9.7 5.7 2.4 12.3 CH,OH 8.3 10.3 9.4 9.7 5.4 2.6 12.2 m-CH, 8.2 10.0 9.2 9.4 5.4 2.4 12.3

”para Substituents unless stated otherwise.

b b b

Unresolved. Appears as AB system: doublet, J = 6.3 Hz.

dJ(op) orJ(mp) = 7.6 Hz; J(o‘o) , J(op) and J(o’p) = 0 Hz

J(H-2, NH)

8.9 8.9 8.9 8.9 8.6 8.8 8.7 8.3 8.3 8.8 8.2

11 .o 8.4 8.6 8.4 8.6 9.0 8.9

J(om)

b

8.4 8.7 8.8

9.0 8.6

8.9 8.9 9.0 9.2 8.8 8.6 8.7 9.0 8.7

c

8.9

d