S1

Supporting Information

for

The Amadori rearrangement as glycoconjugation

method: Synthesis of non-natural C-glycosyl type

glycoconjugates

Katharina Gallas¶, Gerit Pototschnig¶, Florian Adanitsch, Arnold E. Stütz and Tanja M.

Wrodnigg*

Address: Glycogroup, Institute of Organic Chemistry, Technical University Graz,

Stremayrgasse 9, A-8010 Graz, Austria

Email: Tanja M. Wrodnigg – t.wrodnigg@tugraz.at

¶ These authors contributed equally to this work.

* Corresponding author

NMR data of compounds 16, 19, 23–36.

General

NMR spectra were recorded on a Bruker Ultrashield spectrometer at 300.36 and 75.53 MHz,

respectively in methanol-d4 or D2O as indicated. Chemical shifts are listed in delta employing

residual, non-deuterated solvent as the internal standard. Structures of crucial intermediates

were unambiguously assigned by 1D-TOCSY and HSQC experiments.

S2

D-glycero-L-manno/L-gluco-heptopyranose 16a and 16b 1H, 300 MHz, D2O

O

HOOH

HOOH

OH

OH

16

S3

D-glycero-L-manno/L-gluco-heptopyranose 16a and 16b 13C, 75 MHz, D2O

O

HOOH

HOOH

OH

OH

16

S4

1-(N,N-Dibenzylamino)-1-deoxy-αααα-D-galacto-hept-2-ulopyranose (23a) and -furanose (23b), 1H, 300 MHz, MeOH-d4

O

HO OHOH

HO

+O

OH

OHHO

HOOH

NBn2OHNBn2

23a 23b

S5

1-(N,N-Dibenzylamino)-1-deoxy-αααα-D-galacto-hept-2-ulopyranose (23a) and -furanose (23b), 13C, 75 MHz, MeOH-d4

O

HO OH

OH

HO

+O

OH

OHHO

HOOH

NBn2OHNBn2

23a 23b

S6

1-(N-(6-Hydroxyhexyl)amino)-1-deoxy-αααα-d-galacto-hept-2-ulopyranose (24a) and –furanose (24b) 1H, 300 MHz, MeOH-d4

O

HO OHOH

HO

+O

OH

OHHO

HO

OH

NH(CH2)6OHOHNH(CH2)6OH

24a 24b

S7

1-(N-(6-Hydroxyhexyl)amino)-1-deoxy-αααα-d-galacto-hept-2-ulopyranose (24a) and –furanose (24b) 13C, 75 MHz, MeOH-d4

O

HO OHOH

HO

+O

OH

OHHO

HO

OH

NH(CH2)6OHOHNH(CH2)6OH

24a 24b

S8

1-(N-(6-Hydroxyhexyl)amino)-1-N,2-O-carbonyl-1-deoxy-ββββ-D-galacto-hept-2-ulopyranose 26a and -furanose 26b 1H, 300 MHz, MeOH-d4

1253.683

1216.865

1211.323

1173.085

1166.772

1160.464

1156.732

1150.222

1148.015

1145.341

1137.655

1135.411

1127.880

1122.121

1116.191

1114.331

1109.952

1104.806

1099.978

1094.289

1086.690

1083.704

1080.800

1074.332

1067.764

1046.875

1036.979

1014.198

1005.710

1004.119

1002.581

1000.940

999.332

995.870

992.277

985.571

912.571

868.756

489.815

482.914

476.681

471.116

464.503

458.044

440.125

419.071

415.952

O

HO OHOH

HOO

N(CH2)6OH

O O

OHHO

HO

OH

O

N(CH2)6OH

O+

26a 26b

S9

1-(N-(6-Hydroxyhexyl)amino)-1-N,2-O-carbonyl-1-deoxy-ββββ-D-galacto-hept-2-ulopyranose 26a and -furanose 26b 13C, 75 MHz, MeOH-d4

O

HO OHOH

HOO

N(CH2)6OH

O O

OHHO

HO

OH

O

N(CH2)6OH

O+

26a 26b

S10

1-(N-(5-(Methoxycarbonyl)pentyl)amino)-1-deoxy-αααα-D-galacto-hept-2-ulopyranose (25a) and –furanose (25b) 1H, 300 MHz, MeOH-d4

PPM 4.0 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6 1.4 1.2 1.0

1175.366

1170.547

1164.071

1158.043

1151.559

1146.865

1145.473

1141.365

1135.435

1134.903

1132.298

1123.746

1114.897

1109.738

1106.434

1100.033

1096.673

1092.139

1088.756

1084.235

1080.749

1077.645

1075.114

1068.768

1063.625

1053.790

1046.403

1044.708

1037.028

976.744

968.319

966.704

965.084

963.474

961.852

844.098

831.949

826.563

817.362

814.139

809.242

806.771

801.061

798.804

793.759

789.354

786.317

782.003

777.226

774.772

770.099

762.898

758.299

750.918

682.136

677.949

674.814

667.457

477.817

470.387

462.761

455.246

447.751

445.332

437.115

429.752

422.581

397.429

392.794

389.731

386.707

382.870

374.293

357.736

351.026

O

HO OHOH

HO

+O

OH

OHHO

HO

OH

NH(CH2)5CO2MeOHNH(CH2)5CO2Me

25a 25b

S11

1-(N-(5-(Methoxycarbonyl)pentyl)amino)-1-deoxy-αααα-D-galacto-hept-2-ulopyranose (25a) and –furanose (25b) 13C, 75 MHz, MeOH-d4

PPM 94 90 86 82 78 74 70 66 62 58 54 50 46 42 38 34 30

98.1357

82.7965

72.8413

72.7558

72.3858

72.2527

71.5905

71.2552

62.9407

56.0206

52.0818

50.8334

49.9336

49.6501

49.3669

49.2446

49.2271

49.0835

48.9725

48.9426

48.9108

48.8001

48.6027

48.5165

48.2327

34.7830

34.7268

34.5847

30.8513

30.0324

27.8497

27.7432

27.2044

O

HO OHOH

HO

+O

OH

OHHO

HO

OH

NH(CH2)5CO2MeOHNH(CH2)5CO2Me

25a 25b

S12

1-(N-(5-(Methoxycarbonyl)pentyl)amino)-1-N,2-O-carbonyl-1-deoxy-ββββ-D-galacto-hept-2-ulopyranose (27a) and –furanose (27b) 1H,

300 MHz, MeOH-d4

1209.074

1203.173

1200.984

1197.094

1188.316

1178.793

1164.221

1156.802

1154.614

1151.649

1146.989

1144.388

1141.662

1136.457

1125.979

1119.994

1114.475

1106.146

1094.919

1089.569

1082.517

1078.919

1041.105

1031.354

1014.281

1005.701

1004.175

1002.576

1001.043

999.365

995.076

992.053

985.416

978.506

973.733

966.557

718.663

711.301

703.909

517.920

510.577

502.938

495.318

492.261

487.868

484.276

476.802

469.813

428.137

420.347

417.218

413.340

411.232

404.060

O

HO OHOH

HOO

N(CH2)5CO2Me

O O

OHHO

HOOH

O

N(CH2)5CO2Me

O+

27a 27b

S13

1-(N-(5-(Methoxycarbonyl)pentyl)amino)-1-N,2-O-carbonyl-1-deoxy-ββββ-D-galacto-hept-2-ulopyranose (27a) and –furanose (27b) 13C,

75 MHz, MeOH-d4

158.4327

107.3300

104.5388

83.5874

80.2180

75.7478

74.5850

73.7381

71.9974

70.9399

70.7626

63.7771

62.4642

53.4506

52.2688

52.1090

49.9369

49.6532

49.5554

49.5214

49.5119

49.3695

49.2434

49.0857

48.9562

48.8019

48.5182

48.2344

44.3162

44.2806

34.6911

34.6707

27.8660

27.8133

O

HO OHOH

HOO

N(CH2)5CO2Me

O O

OHHO

HOOH

O

N(CH2)5CO2Me

O+

27a 27b

S14

D-glycero-D-galacto/D-talo-heptopyranose 19a and 19b 1H, 300 MHz, D2O

0.998

0.210

0.426

1.085

0.561

PPM 5.00 4.90 4.80 4.70 4.60 4.50 4.40 4.30 4.20 4.10 4.00 3.90 3.80 3.70 3.60 3.50 3.40 3.30

1547.598

1544.241

1540.908

1527.648

1526.612

1442.489

1414.323

1387.527

1386.685

1347.318

1339.480

1216.254

1213.990

1200.682

1197.624

1179.115

1178.627

1175.686

1154.508

1152.633

1151.650

1148.361

1144.496

1138.511

1135.808

1130.433

1123.258

1119.697

1116.049

1113.075

1110.197

1107.281

1096.204

1094.160

1092.272

1091.985

1086.833

1082.104

1076.586

1073.317

1069.876

1063.527

1060.035

1058.110

1051.468

1045.584

1044.056

1036.937

1028.938

1021.432

1019.608

1011.658

O

HOOH

HOOH

OH

OH

19 a and b

S15

D-glycero-D-galacto/D-talo-heptopyranose 19a and 19b 13C, 75 MHz, D2O

PPM 95 93 91 89 87 85 83 81 79 77 75 73 71 69 67 65 63

96.5901

94.0323

92.3434

73.5459

73.0332

71.8321

70.2277

69.4353

69.0700

69.0108

68.7756

68.3358

68.2776

67.7764

63.0065

62.9408

62.7957

O

HOOH

HOOH

OH

OH

19 a and b

S16

1-(N,N-Dibenzylamino)-1-deoxy-αααα-D-manno-hept-2-ulopyranose (28) 1H, 300 MHz, MeOH-d4

PPM 7.2 7.0 6.8 6.6 6.4 6.2 6.0 5.8 5.6 5.4 5.2 5.0 4.8 4.6 4.4 4.2 4.0 3.8 3.6 3.4 3.2 3.0 2.8

7.3117

7.2981

7.2849

7.2694

7.2607

7.2535

7.2417

4.9875

4.1538

3.9524

3.9083

3.8419

3.8318

3.8072

3.8003

3.7891

3.7852

3.7750

3.7558

3.7405

3.7363

3.7192

3.7047

3.6874

3.6742

3.6576

3.6299

3.6089

3.4749

3.4646

3.3772

3.3494

3.3438

3.3384

3.3330

3.3277

3.1409

2.9534

2.9079

2.7924

3.9524

3.9083

3.8419

3.8318

3.8072

3.8003

3.7891

3.7852

3.7750

3.7558

3.7405

3.7363

3.7192

3.7047

3.6874

3.6742

3.6576

3.6299

3.6089

3.4749

3.4646

3.3772

3.3494

3.3438

3.3384

3.3330

3.3277

3.1409

2.9534

2.9079

2.7924

2.7469

O

HO OHOH

HOOH

NBn2

28

S17

1-(N,N-Dibenzylamino)-1-deoxy-αααα-D-manno-hept-2-ulopyranose (28) 13C, 75 MHz, MeOH-d4

PPM 130 126 122 118 114 110 106 102 98 94 90 86 82 78 74 70 66 62 58 54

O

HO OHOH

HOOH

NBn2

28

S18

1-(N-(6-Hydroxyhexyl)amino)-1-deoxy-αααα-D-manno-hept-2-ulose (29) 1H, 300 MHz, MeOH-d4

PPM 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6 1.4

1160.203

1144.939

1139.824

1134.258

1131.942

1127.217

1122.230

1120.808

1110.331

1101.331

1091.555

1083.897

1077.573

1071.130

1014.327

1005.893

1004.232

1002.594

1000.977

959.076

946.449

943.509

920.365

914.160

911.403

905.002

584.160

526.378

520.324

482.602

476.407

470.073

434.044

O

HO OHOH

HOOH

NH(CH2)6OH

29

S19

1-(N-(6-Hydroxyhexyl)amino)-1-deoxy-αααα-D-manno-hept-2-ulose (29) 13C, 75 MHz, MeOH-d4

PPM 92 88 84 80 76 72 68 64 60 56 52 48 44 40 36 32 28

96.2945

75.2374

74.5608

72.5739

67.8860

62.7335

62.5568

54.9609

49.9284

49.6457

49.4290

49.3620

49.0783

48.7945

48.5107

48.2270

33.3555

27.4662

26.7746

O

HO OHOH

HOOH

NH(CH2)6OH

29

S20

1-(N-(6-Hydroxyhexyl)amino)-1-N,2-O-carbonyl-1-deoxy-ββββ-D-manno-pyranose (30) 1H, 300 MHz, MeOH-d4

PPM 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6 1.4 1.2

1135.944

1132.591

1123.688

1118.488

1115.041

1109.512

1103.113

1093.824

1088.328

1086.477

1083.999

1082.009

1073.630

1067.180

1060.605

1052.888

1048.141

1037.629

1030.995

1005.775

995.830

994.192

992.550

990.948

984.772

977.902

971.018

964.128

490.147

482.941

475.931

469.374

463.389

456.965

450.631

432.454

409.861

406.780

384.245

381.460

O

HO OH

OH

HOO

N(CH2)6OH

O

30

S21

1-(N-(6-Hydroxyhexyl)amino)-1-N,2-O-carbonyl-1-deoxy-ββββ-D-manno-pyranose (30) 13C, 75 MHz, MeOH-d4

PPM 96 92 88 84 80 76 72 68 64 60 56 52 48 44 40 36 32 28

77.0664

73.1480

72.5916

67.2968

62.9295

62.2666

54.4060

50.0208

49.7376

49.6397

49.6213

49.5787

49.4534

49.1691

48.8849

48.6006

48.3165

44.7134

33.4872

33.4262

28.1206

27.3783

O

HO OH

OH

HOO

N(CH2)6OH

O

30

S22

1-(N-(5-(Methoxycarbonyl)pentyl)amino)-1-deoxy-αααα-D-manno-hept-2-ulopyranose (31) 1H, 300 MHz, MeOH-d4

PPM 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6

1160.974

1148.400

1143.530

1134.517

1127.027

1125.312

1122.383

1107.074

1020.210

1014.302

1007.867

1005.956

1004.256

1002.667

1001.080

999.583

982.065

974.742

967.427

960.318

950.326

926.973

918.564

911.408

887.759

727.487

720.200

712.904

587.273

537.352

531.905

524.135

516.924

509.446

501.905

494.579

447.801

439.720

432.590

427.166

O

HO OHOH

HOOH

N(CH2)5CO2Me

31

S23

1-(N-(5-(Methoxycarbonyl)pentyl)amino)-1-deoxy-αααα-D-manno-hept-2-ulopyranose (31) 13C, 75 MHz, MeOH-d4

PPM 160 150 140 130 120 110 100 90 80 70 60 50 40 30

175.6750

96.1593

96.1185

96.0809

75.2168

74.6578

72.5274

67.7863

62.4682

55.0595

52.1316

49.9290

49.6454

49.5471

49.3617

49.2889

49.2646

49.1894

49.0780

48.9106

48.7942

48.6080

48.5104

48.2265

47.7365

40.5798

34.4457

28.2792

27.0513

26.9482

O

HO OH

OH

HOOH

N(CH2)5CO2Me

31

S24

1-(N-(5-(Methoxycarbonyl)pentyl)amino)-1-N,2-O-carbonyl-1-deoxy-ββββ-D-manno-hept-2-ulopyranose (32) 1H, 300 MHz, MeOH-d4

PPM 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6 1.4

1156.606

1148.819

1138.399

1135.440

1130.294

1127.318

1119.204

1113.000

1106.301

1083.896

1047.429

1037.041

1014.355

1006.149

1004.283

1002.685

1001.059

999.459

996.297

992.403

989.644

985.475

978.512

943.362

720.315

712.989

705.646

511.973

504.269

496.690

493.084

489.430

485.234

477.831

470.799

427.939

422.310

419.479

415.949

412.685

407.442

O

HO OH

OH

HOO

N(CH2)5COOMe

O

32

S25

1-(N-(5-(Methoxycarbonyl)pentyl)amino)-1-N,2-O-carbonyl-1-deoxy-ββββ-D-manno-hept-2-ulopyranose (32) 13C, 75 MHz, MeOH-d4

PPM 160 150 140 130 120 110 100 90 80 70 60 50 40 30

175.8586

158.0638

103.7579

77.2452

73.2462

72.6996

67.4647

62.5605

54.3404

52.1021

49.9454

49.6617

49.3779

49.2613

49.1979

49.0941

48.9593

48.9270

48.8104

48.5267

48.2430

44.5138

34.6467

27.8596

27.0334

O

HO OH

OH

HOO

N(CH2)5COOMe

O

32

S26

1-(N,N-Dibenzylamino)-3-acetamino-1,2-dideoxy-αααα,D-gluco-hept-2-ulopyranose (33a) and –furanose (33b) 1H, 300 MHz, MeOH-d4

6.878

3.453

2.187

3.258

3.310

2.998

PPM 4.0 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6

1261.565

1251.431

1238.245

1161.676

1159.198

1156.959

1153.918

1150.890

1147.098

1144.648

1141.504

1139.153

1136.728

1131.452

1124.805

1119.977

1116.538

1109.940

1105.292

1102.976

1096.994

1095.759

1088.516

1086.835

1081.551

1039.473

1032.723

1019.761

1014.335

1005.927

1004.299

1002.705

1001.103

999.558

794.470

788.681

779.840

616.810

606.557

602.918

596.561

488.684

O

HO NHAcOH

HOOH

NBn2 O

OH

NHAcHO

HOOH

NBn2

33a 33b

+

S27

1-(N,N-Dibenzylamino)-3-acetamino-1,2-dideoxy-αααα,D-gluco-hept-2-ulopyranose (33a) and –furanose (33b) 13C, 75 MHz, MeOH-d4

PPM 160 150 140 130 120 110 100 90 80 70 60 50 40 30

141.0437

140.1024

132.8707

131.1222

130.6504

130.5260

130.3568

130.2496

129.6439

129.3361

128.4955

128.0596

98.2584

74.4304

73.8244

72.2186

62.9686

60.4703

57.4311

56.5703

56.4964

51.9922

50.0730

49.7896

49.5062

49.2226

48.9390

48.6553

48.3715

40.5470

23.0481

O

HO NHAcOH

HOOH

NBn2 O

OH

NHAcHO

HOOH

NBn2

33a 33b

+

S28

1-(N-(6-Hydroxyhexyl)amino)-3-acetamino-1,2-dideoxy-αααα-D-gluco-hept-2-ulopyranose (34) 1H, 300 MHz, MeOH-d4

4.582

4.542

3.251

1.036

1.876

1.083

1.000

2.508

5.062

PPM 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6 1.4

1137.754

1134.054

1126.090

1123.520

1081.679

1075.235

1068.698

1053.241

1051.027

1044.305

1037.825

1035.403

1014.285

1005.820

1004.221

1002.617

1001.021

999.443

845.325

833.030

808.663

801.276

793.897

788.383

776.060

657.238

615.969

578.858

474.161

468.656

426.542

423.404

420.085

O

HO NHAcOH

HOOH

NH(CH2)6OH

34

S29

1-(N-(6-Hydroxyhexyl)amino)-3-acetamino-1,2-dideoxy-αααα-D-gluco-hept-2-ulopyranose (34) 13C, 75 MHz, MeOH-d4

PPM 160 150 140 130 120 110 100 90 80 70 60 50 40 30

98.1194

74.1611

73.4722

71.7321

62.9173

62.5114

56.0525

55.9567

50.8665

49.9719

49.6889

49.4053

49.1216

48.8379

48.5541

48.2704

33.5839

29.9953

28.0400

26.8168

22.7352

O

HO NHAcOH

HOOH

NH(CH2)6OH

34

S30

1-(N-(5-(Methoxycarbonyl)pentyl)amino)-2-acetamino-1,2-dideoxy-αααα-D-gluco-hept-2-ulopyranose (35) 1H, 300 MHz, MeOH-d4

0.995

1.144

4.150

2.809

3.517

4.224

2.334

4.687

2.313

PPM 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6

O

HO NHAcOH

HOOH

NH(CH2)5COOMe

35

S31

1-(N-(5-(Methoxycarbonyl)pentyl)amino)-2-acetamino-1,2-dideoxy-αααα-D-gluco-hept-2-ulopyranose (35) 13C, 75 MHz, MeOH-d4

PPM 94 90 86 82 78 74 70 66 62 58 54 50 46 42 38 34 30 26

96.7466

74.5095

72.1755

71.4365

61.7013

56.0322

54.1367

52.1093

49.9016

49.7566

49.6179

49.5241

49.4712

49.3341

49.1933

49.0503

48.8933

48.7665

48.6175

48.4827

48.1990

34.4281

26.9674

26.7453

25.4152

22.5047

O

HO NHAcOH

HOOH

NH(CH2)5COOMe

35

S32

1-N-[(5S)-(tert-Butoxylcarbonylamino)-5-(methoxycarbonyl)pentyl]amino-3-acetamino-1,2-dideoxy-αααα-D-gluco-hept-2-ulopyranose (36) 1H, 300 MHz, MeOH-d4

PPM 4.0 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6 1.4

4.1320

4.1206

4.1050

3.8225

3.7814

3.7748

3.7647

3.7546

3.7406

3.6926

3.6708

3.6472

3.6238

3.6004

3.5017

3.4733

3.4442

3.3769

3.3489

3.3435

3.3381

3.3328

3.3274

2.7649

2.7240

2.6351

2.6121

2.5884

2.5457

2.5088

2.5061

2.0858

2.0497

1.8337

1.8220

1.7974

1.7802

1.7548

1.7457

1.7166

1.6948

1.6687

1.6510

1.6398

1.6201

1.5934

1.5700

1.5465

1.5288

O

HO NHAcOH

HOOH

NH(CH2)4O

OMe

NH O

O

36

S33

1-N-[(5S)-(tert-Butoxylcarbonylamino)-5-(methoxycarbonyl)pentyl]amino-3-acetamino-1,2-dideoxy-αααα-D-gluco-hept-2-ulopyranose (36) 13C, 75 MHz, MeOH-d4

PPM 160 150 140 130 120 110 100 90 80 70 60 50 40 30

174.1236

158.1771

98.3104

98.2767

80.6013

74.1004

73.6651

71.7705

62.6079

58.3844

56.2606

56.0407

55.8650

55.0858

54.9977

52.6748

50.7030

49.9598

49.6760

49.3922

49.2238

49.1084

48.9412

48.8246

48.6238

48.5410

48.2574

32.4966

29.9187

28.8213

24.5289

22.7436

O

HO NHAcOH

HOOH

NH(CH2)4O

OMe

NH O

O

36