First Total Synthesis of (+)-Broussonetine W: Glycosidase ... · S1 First Total Synthesis of...
72
S1 First Total Synthesis of (+)-Broussonetine W: Glycosidase Inhibition of Natural Product & Analogs Ying-Ying Song, a,e Kyoko Kinami, b Atsushi Kato, *,b Yue-Mei Jia, a Yi-Xian Li, a George W. J. Fleet c,d and Chu-Yi Yu *,a,d a Beijing National Laboratory of Molecular Science (BNLMS), CAS Key Laboratory of Molecular Recognition and Function, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China b Department of Hospital Pharmacy, University of Toyama, 2630 Sugitani, Toyama 930-0194, Japan c Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Mansfield Road, Oxford OX1 3TA, U.K. d National Engineering Research Center for Carbohydrate Synthesis, Jiangxi Normal University, Nanchang, 330022, China e University of the Chinese Academy of Sciences, Beijing 100049, China Contents 1. Comparison of 13 C NMR of the synthetic broussonetine W (4) with natural products.......................................S2 2. Table for all the new compounds………….……………………………………………………………………S3 3. Copies of 1 H , 13 C and 2D NMR spectra for all the new compounds..................................................................S6 4. Results of the bioassay………………………………………………………………………………………...S70 Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry. This journal is © The Royal Society of Chemistry 2016
Transcript of First Total Synthesis of (+)-Broussonetine W: Glycosidase ... · S1 First Total Synthesis of...
S1
First Total Synthesis of (+)-Broussonetine W: Glycosidase Inhibition of
Natural Product & Analogs
Ying-Ying Song,a,e Kyoko Kinami,b Atsushi Kato,*,b Yue-Mei Jia,a Yi-Xian Li,a George W. J. Fleetc,d and
Chu-Yi Yu*,a,d
a Beijing National Laboratory of Molecular Science (BNLMS), CAS Key Laboratory of Molecular Recognition and
Function, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China b Department of Hospital Pharmacy, University of Toyama, 2630 Sugitani, Toyama 930-0194, Japan c Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Mansfield Road, Oxford OX1
3TA, U.K. d National Engineering Research Center for Carbohydrate Synthesis, Jiangxi Normal University, Nanchang, 330022,
China e University of the Chinese Academy of Sciences, Beijing 100049, China
Contents
1. Comparison of 13C NMR of the synthetic broussonetine W (4) with natural products.......................................S2
2. Table for all the new compounds………….……………………………………………………………………S3
3. Copies of 1H , 13C and 2D NMR spectra for all the new compounds..................................................................S6
4. Results of the bioassay………………………………………………………………………………………...S70
Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry.This journal is © The Royal Society of Chemistry 2016
S2
1. Comparison of 13C NMR of synthetic broussonetine W (4) with natural
product.
Table 1. Comparison of the synthetic broussonetine W (4) with natural product
W (lit.)a W (4)b
Pyrrolidine
moiety
CH2OH 63.45 62.99 -0.46 2 64.97 65.21 0.24 3 80.07 79.60 -0.47 4 84.06 83.53 -0.53 5 62.74 62.85 0.11
Octyl
moiety
1’ 35.43 34.80 -0.63 2’ 27.23 27.23 0.00 3’ 30.12 30.14 0.02 4’
29.74, 29.70 29.82, 29.82 0.08, 0.12 5’ 6’ 28.98 29.09 0.11 7’ 29.96 30.09 0.13
Cyclohexenone
moiety
1’’ 139.79 139.91 0.12 2’’ 198.81 198.81 0.00 3’’ 38.81 38.94 0.13 4’’ 23.43 23.57 0.14 5’’ 26.09 26.22 0.13 6’’ 145.09 145.15 0.06
a) δ in pyridine-d5 at 125 MHz. b) δ in pyridine-d5 at 126 MHz.
S3
2. Table for all the new compounds.
Experimental procedures and characterization data for all the new compounds see the general procedures in essay
for the details.
Table 1. New Compounds in the total synthesis of (4, ent-4, 3-epi-4)
Synthesis of 4 Synthesis of ent-4 Synthesis of 3-epi-4
Cyclic Nitrones:
(Starting Materials)
N
OBn
BnO
BnO
O
D-lyxo-nitrone(3-epi-10)
Step1-3:
Grignard addition
Reduction
Protection of amino group
87% yield in 3 steps starting
from 10
98% yield in 3 steps starting
from ent-10
66% yield in 3 steps starting
from 3-epi-10
Step 4:
Hydroboration-Oxidation
73% yield derived from olefin
17
93% yield derived from olefin
ent-17
100% yield derived from
olefin 3-epi-17
Step 5:
Appel reaction
97% yield derived from
alcohol 18
85% yield derived from
alcohol ent-18
88% yield derived from
alcohol 3-epi-18
Step 6:
α-alkylation of imine
(N-cyclohexylidenecyclohex
anamine 14)
73% yield derived from
bromide 15
71% yield derived from
bromide ent-15
60% yield derived from
alcohol 3-epi-15
Step 7:
10% Pd/C-catalyzed
hydrogenolysis reaction
96% yield derived from
ketone 19
84% yield derived from
ketone ent-19
75% yield derived from
ketone 3-epi-19
S4
Step 8:
Protection of the amino and
hydroxyl groups with Ac2O
95% yield derived from
ketone 20
94% yield derived from
ketone ent-20
76% yield derived from
ketone 3-epi-20
Step 9-10:
Bromination-elimination of
ketone
N
OAc
AcO
AcO22
5
OAc
83% yield in 2 steps derived
from ketone 21
85% yield in 2 steps derived
from ketone ent-21
78% yield in 2 steps derived
from ketone 3-epi-21
Step 11:
Removal of acetyl groups
92% yield derived from
compound 22
78% yield derived from
compound ent-22
89% yield derived from
compound 3-epi -22
Total Yield In 10 Steps: 31% 29% 14%
Table 2. New Compounds in the total synthesis of (ent-3-epi-4, ent-4a, ent-4b)
Synthesis of ent-3-epi-4 Synthesis of ent-4a Synthesis of ent-4b
Cyclic Nitrones:
(Starting Materials)
Step1-3:
Grignard addition
Reduction
Protection of amino group
76% yield in 3 steps starting
from ent-3-epi-10
84% yield in 3 steps starting
from ent-10
93% yield in 3 steps starting
from ent-10
Step 4:
Hydroboration-Oxidation
73% yield derived from olefin
ent-3-epi-17
N
OBn
BnO
BnO
OH
ent-18a
Cbz
4
91% yield derived from olefin
ent-17a
91% yield derived from olefin
ent-17b
S5
Step 5:
Appel reaction
96% yield derived from
alcohol ent-3-epi-18
92% yield derived from
alcohol ent-18a
93% yield derived from
alcohol ent-18b
Step 6:
α-alkylation of imine
(N-cyclohexylidenecyclohex
anamine 14)
71% yield derived from
bromide ent-3-epi-15
68% yield derived from
bromide ent-15a
71% yield derived from
alcohol ent-15b
Step 7:
10% Pd/C-catalyzed
hydrogenolysis reaction
79% yield derived from
ketone ent-3-epi-19
82% yield derived from
ketone ent-19a
83% yield derived from
ketone ent-19b
Step 8:
Protection of the amino and
hydroxyl groups with Ac2O
94% yield derived from
ketone ent-3-epi-20
93% yield derived from
ketone ent-20a
90% yield derived from
ketone ent-20b
Step 9-10:
Bromination-elimination of
ketone
83% yield in 2 steps derived
from ketone ent-3-epi-21
74% yield in 2 steps derived
from ketone ent-21a
74% yield in 2 steps derived
from ketone ent-21b
Step 11:
Removal of acetyl groups
77% yield derived from
compound ent-3-epi-22
86% yield derived from
compound ent-22a
74% yield derived from
compound ent-22b
Total Yield In 10 Steps: 18% 23% 23%
S6
3. Copies of 1H , 13C and 2D NMR spectra of new compounds.
9 8 7 6 5 4 3 2 1 0 ppm
1.215
1.227
1.241
1.253
1.262
1.278
1.287
1.301
1.315
1.328
1.401
1.415
1.782
1.929
1.943
1.956
1.970
3.078
3.088
3.093
3.099
3.104
3.115
3.444
3.455
3.464
3.492
3.511
3.524
3.715
3.719
3.861
3.865
4.363
4.369
4.386
4.393
4.397
4.421
4.468
4.491
4.841
4.861
4.891
4.925
5.693
5.714
5.727
5.748
7.177
7.188
7.207
7.215
7.232
7.239
7.248
7.260
7.261
6.272
1.045
1.029
2.006
0.962
1.000
0.969
1.931
0.973
6.098
1.955
1.899
15.373
190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 ppm
26.542
28.988
29.429
33.873
68.480
70.164
71.765
71.813
73.459
84.748
86.852
114.364
127.747
127.806
127.841
127.985
128.113
128.464
128.476
128.498
138.195
138.311
139.216
1H NMR, 500 MHz, CDCl3
13C NMR, 126 MHz, CDCl3
COSY oof compound 16
H3
H4 H
(H5
(H3, H4)
H3
H4
S7
H5H2
H6
5, H1’)
(H5, H4)
H5H2
H6
(H5, H3)
NOESY, 5
1H-1
(H5, H6)
500 MHz, CDC
H COSY, 500 M
Cl3
MHz, CDCl3
S8
9 8 7 6 5 4 3 2 1 0 ppm
1.147
1.158
1.206
1.260
1.272
1.353
1.943
1.957
2.000
2.012
3.475
3.487
3.491
3.720
3.722
3.753
3.776
3.792
3.814
3.832
3.855
4.133
4.157
4.329
4.344
4.355
4.370
4.396
4.427
4.452
4.474
4.573
4.605
4.628
4.651
4.914
4.934
4.959
4.977
4.993
5.021
5.046
5.151
5.176
5.191
5.216
5.759
5.779
7.175
7.188
7.200
7.214
7.225
7.266
7.288
7.301
7.322
7.335
6.429
1.745
2.545
1.000
2.524
0.500
1.520
0.515
2.410
2.154
1.527
1.957
1.007
1.022
0.951
19.889
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
28.771
28.939
29.001
30.170
31.465
33.739
33.791
62.680
63.010
64.766
65.091
66.893
66.953
67.898
68.850
70.936
71.115
71.266
73.064
73.131
82.080
83.233
83.415
84.493
114.341
114.388
127.610
127.671
127.718
127.755
127.799
127.847
127.865
128.091
128.168
128.256
128.427
128.464
128.530
128.594
136.694
136.741
137.752
137.789
138.002
138.059
138.400
138.645
139.078
139.186
154.328
154.757
1H NMR, 500 MHz, CDCl3
13C NMR, 126 MHz, CDCl3
S9
9 8 7 6 5 4 3 2 1 0 ppm
1.156
1.165
1.189
1.258
1.291
1.452
1.462
1.488
1.510
1.531
1.673
3.471
3.501
3.506
3.564
3.586
3.605
3.627
3.746
3.793
3.814
4.156
4.316
4.341
4.350
4.355
4.369
4.390
4.417
4.430
4.440
4.458
4.481
4.567
4.601
4.607
4.621
4.641
4.661
5.012
5.053
5.144
5.185
5.226
7.179
7.193
7.204
7.212
7.218
7.231
7.245
7.263
7.272
7.285
7.306
7.321
7.333
7.345
8.507
2.365
1.893
0.535
0.997
1.958
2.531
0.473
1.496
0.529
2.409
2.109
1.482
0.971
1.000
19.897
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
137.715
25.621
25.676
26.439
26.527
29.248
29.297
30.046
31.470
32.759
62.676
63.018
64.806
65.095
66.986
67.834
68.782
70.916
71.093
71.241
73.054
73.118
82.020
83.171
83.397
84.471
127.593
127.664
127.697
127.745
127.799
128.118
128.243
128.420
128.450
128.519
128.589
136.613
137.741
137.958
138.016
138.332
138.572
154.438
154.839
1H NMR, 300 MHz, CDCl3
13C NMR, 75 MHz, CDCl3
S10
9 8 7 6 5 4 3 2 1 0 ppm
1.150
1.283
1.329
1.374
1.636
1.644
1.668
1.689
1.711
1.735
1.761
1.786
1.812
1.836
2.027
3.322
3.345
3.368
3.390
3.443
3.474
3.508
3.709
3.722
3.738
3.751
3.789
3.810
3.856
4.051
4.122
4.135
4.161
4.319
4.349
4.359
4.376
4.392
4.420
4.438
4.460
4.479
4.571
4.601
4.618
4.641
4.659
5.012
5.053
5.145
5.189
5.230
7.196
7.218
7.260
7.286
7.321
7.334
8.492
0.663
3.170
0.549
1.827
0.966
2.472
0.486
1.481
0.512
2.411
2.048
1.479
0.980
1.000
19.663
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
28.551
28.677
29.046
29.230
30.123
31.437
32.802
32.856
34.019
34.070
62.684
63.031
64.751
65.081
66.889
66.936
67.897
68.846
70.941
71.122
71.263
73.067
73.130
82.077
83.215
83.471
84.522
127.602
127.655
127.693
127.734
127.780
127.841
128.086
128.146
128.232
128.409
128.447
128.523
128.574
136.726
137.747
138.002
138.391
138.634
154.317
154.731
1H NMR, 300 MHz, CDCl3
13C NMR, 75 MHz, CDCl3
S11
5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 ppm
-0.000
1.022
1.028
1.054
1.136
1.144
1.153
1.167
1.175
1.183
1.198
1.205
1.213
1.228
1.235
1.241
1.260
1.291
1.298
1.323
1.330
1.354
1.365
1.396
1.429
1.454
1.461
1.585
1.632
1.640
1.647
1.695
1.723
1.744
1.752
1.784
1.835
1.850
1.867
1.881
1.896
2.239
2.256
2.271
2.281
2.296
2.321
2.339
2.355
2.618
3.262
3.273
3.288
3.299
3.309
3.325
5.471
7.287
4.391
4.259
1.000
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
25.132
25.687
26.250
27.638
27.937
29.117
34.205
40.309
57.905
170.829
1H NMR, 400 MHz, CDCl3
13C NMR, 101 MHz, CDCl3
S12
9 8 7 6 5 4 3 2 1 0 ppm
1.083
1.110
1.164
1.179
1.276
1.535
1.544
1.563
1.571
1.577
1.590
1.599
1.610
1.621
1.637
1.662
1.674
1.921
1.927
1.962
1.975
1.985
2.146
2.165
2.187
2.284
3.394
3.670
3.718
3.739
4.077
4.278
4.295
4.319
4.341
4.366
4.382
4.407
4.488
4.520
4.529
4.539
4.579
4.936
4.977
5.068
5.105
7.095
7.102
7.119
7.138
7.144
7.162
7.184
7.208
7.225
7.245
7.255
12.456
6.063
2.567
2.053
0.999
0.968
2.512
0.526
1.505
0.581
2.018
2.532
1.471
0.972
0.984
20.000
2030405060708090100110120130140150160170180190200210 ppm
28.139
29.319
29.478
29.574
29.785
30.212
66.913
31.538
33.923
33.951
42.062
50.862
62.682
63.009
64.784
65.102
66.873
67.892
68.844
70.933
71.093
71.240
73.046
73.110
82.105
83.232
83.421
84.502
127.591
127.650
127.696
127.728
127.773
127.823
128.092
128.232
128.406
128.441
128.515
128.572
136.706
137.754
138.005
138.061
138.386
138.631
154.316
154.759
213.631
213.703
21222324252627282930313233343536373839 ppm
29.523
24.924
26.658
27.262
28.139
29.319
29.478
29.574
29.785
30.212
31.538
33.923
33.951
1H NMR, 300 MHz, CDCl3
13C NMR, 75 MHz, CDCl3
S13
9 8 7 6 5 4 3 2 1 0 ppm
1.241
1.268
1.289
1.307
1.393
1.413
1.463
1.468
1.504
1.542
1.572
1.699
1.715
1.742
1.753
1.785
1.807
1.822
1.832
1.853
1.915
1.927
2.110
2.117
2.130
2.140
2.171
2.184
2.190
2.204
2.215
2.220
2.403
2.419
2.436
2.457
3.000
3.019
3.125
3.140
3.160
3.371
3.379
3.385
3.390
3.395
3.671
3.690
3.710
3.730
3.755
3.769
3.793
3.807
3.847
3.869
3.890
4.965
14.693
4.247
0.963
2.055
2.430
1.000
1.013
3.084
1.117
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
25.784
27.596
28.211
29.274
30.489
30.590
30.704
30.809
34.757
35.291
42.780
51.753
62.689
62.904
64.577
79.159
83.110
216.076
1H NMR, 300 MHz, MeOD
13C NMR, 75MHz, MeOD
S14
9 8 7 6 5 4 3 2 1 0 ppm
1.728
1.744
1.763
1.776
1.831
1.959
2.002
2.075
2.086
2.089
2.123
2.191
2.233
2.247
2.263
2.279
2.294
2.308
2.350
2.382
3.727
3.752
3.971
3.995
4.030
4.056
4.071
4.079
4.106
4.172
4.196
4.221
4.265
4.295
4.382
4.388
4.407
4.414
4.492
4.499
4.517
4.524
5.074
5.088
5.147
13.563
5.017
12.855
4.160
0.502
1.269
1.035
0.408
0.500
0.937
0.918
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
26.194
26.304
27.199
28.102
29.338
29.396
29.472
29.676
29.722
30.055
33.285
33.878
33.911
33.983
42.008
42.039
50.798
60.492
62.332
62.786
62.962
65.419
65.775
76.349
77.518
77.791
78.619
169.075
169.217
169.459
169.619
169.655
169.897
170.928
170.999
213.589
213.693
1H NMR, 400 MHz, CDCl3
1H NMR, 101 MHz, CDCl3
S15
9 8 7 6 5 4 3 2 1 0 ppm
1.311
1.355
1.367
1.382
1.629
1.638
1.660
1.673
1.815
1.840
1.854
1.933
1.953
1.974
1.995
2.016
2.064
2.089
2.097
2.105
2.114
2.138
2.207
2.340
2.356
2.377
2.390
2.412
2.434
3.736
3.744
3.774
3.782
3.980
4.010
4.045
4.077
4.085
4.095
4.178
4.210
4.243
4.273
4.284
4.306
4.316
4.393
4.403
4.424
4.434
4.503
4.514
4.535
4.547
5.091
5.102
5.162
6.703
11.167
1.771
2.044
13.831
3.742
0.500
2.440
0.383
0.482
0.901
0.886
0.878
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
23.192
26.074
26.156
26.269
28.546
28.642
29.240
29.284
29.328
29.507
29.578
30.015
33.235
38.628
60.452
62.290
62.748
62.925
65.377
65.727
76.313
77.483
77.752
78.576
139.877
139.911
144.918
144.988
169.032
169.162
169.412
169.560
169.603
169.853
170.873
170.941
199.566
199.590
1H NMR, 300 MHz, CDCl3
N
OAcAcO
AcO
O
22
Ac
13C NMR, 75 MHz, CDCl3
S16
9 8 7 6 5 4 3 2 1 0 ppm
1.395
1.539
1.629
1.638
1.750
1.762
1.775
1.787
1.802
2.035
2.047
2.055
2.082
2.135
2.143
2.187
2.202
2.227
2.241
2.256
2.368
2.381
2.394
3.597
3.607
3.867
3.874
4.206
4.228
4.239
4.259
4.265
4.280
4.417
4.429
4.443
4.691
4.704
4.716
6.572
6.909
6.872
2.294
1.279
1.367
2.878
0.975
1.597
1.749
1.544
0.772
0.719
1.668
1.000
0.956
0.966
2.723
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
23.566
26.223
27.234
29.094
29.817
30.088
30.143
34.796
38.935
62.849
62.992
65.207
79.596
83.531
139.908
145.153
198.810
1H NMR, 500 MHz, Pyr.
1H NMR, 126 MHz, Pyr.
COSY o
HSQC o
of the synthet
of the synthet
7.5
tic broussone
tic broussonti
6.57.0
H-6’’
etine W (4)
ine W (4)
5.56.0
H-6’’, H-5’’
C-6’’,
S17
4.55.0
H-3 H-4
CH
H-6’’
33.54.0
H2OH
H-2 H-5
H-3’’, H
HSQC, 500
2.2.5.0
H-3’’
H-7’
HH-5’’
H-4’’
H-4’’, H-5’
C-3’’, H-3’’
C-5’’, H-5’’
H-7’
COSY, 500M, Py
0M, Pyr.
1.01.50
H-1’ ’
H-2’
H-4’’
H-6’
’’
C-4’’, H-4’’
yr.
ppm
0 ppm
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0
6.5
7.0
7.5
HMBC
of the syntheetic broussone
HNHOH2C
HO
2
3
etine W (4)
HN
OH4
5 1'
broussonetin
HMBC, 500M
C-2’’, H-6’’
S18
O
7' 2''
H 6''
ne W (4)
H H
M, Pyr.
4''
HHH
H
C-2’’, H-
C-2’’, H
C-1’’, H
C-6’’, H
HMBC
3’’
H-7’
H-7’
C-1
H-7’
H-3’’
H-7’
H-5’’ H
C-2’’, H-4’’
1’’, H-5’’
C-6’’, H-5’’
C-6’’, H-4’’
H-4’’
S19
9 8 7 6 5 4 3 2 1 0 ppm
1.310
1.322
1.332
1.344
1.349
1.356
1.372
1.386
2.012
2.027
3.154
3.170
3.525
3.562
3.580
3.594
3.784
3.788
3.793
3.798
3.930
3.935
3.937
3.943
4.437
4.456
4.461
4.467
4.535
4.562
4.909
4.911
4.913
4.929
4.931
4.933
4.959
4.963
4.993
4.997
5.784
5.797
7.244
7.259
7.264
7.268
7.274
7.278
7.285
7.293
7.297
7.302
7.303
7.308
7.317
7.328
7.331
7.334
6.372
1.101
1.025
2.049
1.000
1.007
1.020
1.993
1.004
6.349
0.930
0.962
0.958
0.819
15.517
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
26.547
28.991
29.438
33.874
68.444
70.102
70.202
71.747
71.803
73.451
84.729
86.850
114.365
127.738
127.796
127.833
127.980
128.109
128.458
128.471
128.491
138.197
138.214
138.315
139.210
1H NMR, 500 MHz, CDCl3
13C NMR, 126 MHz, CDCl3
H
H
3 H2
H4 H6
(H
H5
(H5, H
H5, H3)
S20
COSY
H5H3 H2
H4 H6
NOE
H6)
Y, 500 MHz, C
ESY, 500MHz,
CDCl3
, CDCl3
S21
9 8 7 6 5 4 3 2 1 ppm
1.15
51.
257
1.32
21.
345
1.93
11.
954
1.98
52.
007
3.44
53.
477
3.50
93.
725
3.75
43.
793
3.81
63.
838
4.16
34.
283
4.30
54.
328
4.33
44.
357
4.37
54.
407
4.42
94.
443
4.46
74.
555
4.58
94.
607
4.62
94.
647
4.90
44.
937
4.99
85.
040
5.14
15.
183
5.22
55.
731
5.75
47.
181
7.23
97.
268
7.29
8
6.34
2
1.62
7
2.59
0
1.02
22.
556
0.54
11.
553
3.00
82.
121
1.59
53.
049
1.05
0
1.00
0
20.2
56
190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 ppm
26.410
26.452
28.711
28.879
28.945
30.108
31.401
33.688
33.739
62.615
62.943
64.703
65.028
66.837
66.898
67.823
68.775
70.868
71.048
71.201
73.002
73.070
81.998
83.155
83.333
84.411
114.300
114.344
127.559
127.618
127.665
127.699
127.747
127.798
128.038
128.114
128.203
128.374
128.411
128.480
128.538
136.626
136.673
137.685
137.723
137.935
137.992
138.333
138.575
139.020
139.127
154.268
154.698
1H NMR, 300 MHz, CDCl3
13C NMR, 75 MHz, CDCl3
S22
9 8 7 6 5 4 3 2 1 0 ppm
1.159
1.280
1.290
1.458
1.472
1.486
1.501
1.514
3.455
3.475
3.494
3.544
3.558
3.571
3.585
3.716
3.724
3.733
3.757
3.779
3.794
3.815
3.835
3.858
4.137
4.159
4.319
4.344
4.361
4.387
4.416
4.445
4.468
4.566
4.597
4.620
4.642
5.013
5.016
5.038
5.041
5.144
5.146
5.168
5.171
5.185
5.188
5.210
5.213
7.179
7.186
7.205
7.234
7.252
7.278
7.293
7.310
8.339
2.586
1.036
1.058
0.600
0.973
1.972
2.506
0.463
1.488
0.491
2.377
2.174
1.478
0.973
1.000
19.737
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
26.381
26.445
29.186
29.258
29.986
31.407
32.688
32.706
62.588
62.789
62.926
64.702
65.013
66.834
66.861
67.764
68.719
70.826
71.004
71.147
72.959
73.025
81.945
83.076
83.322
84.381
127.513
127.578
127.621
127.661
127.707
127.746
127.984
128.026
128.144
128.327
128.360
128.430
128.493
136.571
137.634
137.668
137.879
137.935
138.253
138.495
154.266
154.686
1H NMR, 500 MHz, CDCl3
13C NMR, 126 MHz, CDCl3
S23
9 8 7 6 5 4 3 2 1 0 ppm
1.141
1.262
1.281
1.724
1.740
1.776
1.791
3.307
3.321
3.328
3.335
3.342
3.356
3.478
3.791
3.812
4.162
4.166
4.322
4.336
4.347
4.351
4.366
4.387
4.420
4.444
4.468
4.571
4.595
4.602
4.618
4.627
4.642
5.012
5.017
5.036
5.042
5.149
5.174
5.196
5.221
7.165
7.168
7.181
7.194
7.208
7.230
7.243
7.250
7.253
7.277
7.285
7.295
7.309
7.317
7.325
7.334
8.317
0.595
3.203
0.513
1.865
0.998
2.522
0.517
1.494
0.499
2.413
2.122
1.490
1.001
1.000
19.806
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
28.936
29.127
30.015
31.330
32.688
32.745
33.950
34.001
62.580
62.933
64.645
64.981
66.785
66.842
67.766
68.718
70.822
71.002
71.146
72.953
73.020
81.929
83.066
83.322
84.374
127.509
127.567
127.603
127.653
127.696
127.747
127.764
127.999
128.054
128.143
128.314
128.322
128.355
128.407
128.438
128.485
136.584
136.618
137.634
137.666
137.887
137.937
138.277
138.515
154.209
154.622
1H NMR, 500 MHz, CDCl3
13C NMR, 126 MHz, CDCl3
S24
9 8 7 6 5 4 3 2 1 0 ppm
1.080
1.111
1.164
1.179
1.278
1.309
1.541
1.565
1.587
1.614
1.625
1.654
1.671
1.686
1.742
1.753
1.768
1.926
1.950
1.989
2.156
2.168
2.188
2.218
2.274
2.285
3.395
3.718
3.740
4.077
4.244
4.275
4.292
4.320
4.346
4.375
4.403
4.491
4.523
4.543
4.554
4.573
4.941
4.972
5.071
5.107
5.139
7.117
7.137
7.158
7.167
7.182
7.208
7.227
7.237
7.244
7.255
7.266
12.875
6.382
2.674
2.089
1.028
1.000
2.561
0.496
1.537
0.519
2.015
2.635
1.513
1.006
1.013
20.466
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
29.309
29.467
29.513
29.567
29.781
30.204
31.528
33.945
42.057
50.850
62.669
62.995
64.769
65.090
66.858
66.898
67.878
68.831
70.920
71.079
71.229
73.034
73.100
82.087
83.218
83.404
84.487
127.581
127.641
127.687
127.721
127.766
127.816
128.035
128.084
128.225
128.398
128.432
128.509
128.566
136.722
137.775
137.992
138.050
138.374
138.619
154.299
154.743
213.610
213.684
1H NMR, 400 MHz, CDCl3
2122232425262728293031323334 ppm
33.915
24.927
26.651
27.254
28.134
29.310
29.468
29.514
29.567
29.781
30.204
31.529
33.946
13C NMR, 101 MHz, CDCl3
S25
9 8 7 6 5 4 3 2 1 0 ppm
1.705
1.724
1.732
1.740
1.760
1.835
1.844
1.857
1.867
1.894
2.025
2.043
2.094
2.127
2.168
2.285
2.308
2.319
2.330
2.350
2.365
2.405
2.952
2.970
2.985
3.003
3.082
3.097
3.108
3.123
3.621
3.638
3.654
3.666
3.699
3.708
3.727
3.737
3.793
3.809
3.825
14.493
4.012
1.024
1.847
2.259
0.765
0.792
2.610
1.000
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
26.654
28.398
29.075
30.150
31.328
31.455
31.516
31.668
35.240
36.171
43.653
52.624
63.121
63.922
65.546
79.666
83.594
217.026
1H NMR, 400 MHz, MeOD
13C NMR, 101 MHz, MeOD
S26
9 8 7 6 5 4 3 2 1 0 ppm
1.283
1.360
1.397
1.662
1.758
1.847
2.015
2.098
2.102
2.136
2.205
2.247
2.269
2.356
2.395
3.732
3.770
3.979
4.006
4.040
4.074
4.127
4.178
4.211
4.243
4.274
4.306
4.393
4.424
4.499
4.531
5.089
5.159
7.303
12.823
5.532
12.682
4.458
0.550
1.477
1.085
0.410
0.500
0.943
0.887
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
24.836
24.874
26.169
26.277
27.172
28.078
29.311
29.370
29.448
29.650
29.695
30.032
33.260
33.886
33.960
42.013
50.768
60.464
62.307
62.760
62.940
65.389
65.741
76.325
77.499
77.770
78.598
169.033
169.177
169.419
169.586
169.838
170.878
170.955
213.511
213.614
1H NMR, 300 MHz, CDCl3
15202530354045 ppm
21.063
33.855
41.983
20.753
20.893
20.957
21.028
22.845
24.836
24.874
26.169
26.277
27.172
28.078
29.311
29.370
29.448
29.650
29.695
30.032
33.260
33.886
33.960
42.013
13C NMR, 75 MHz, CDCl3
S27
9 8 7 6 5 4 3 2 1 0 ppm
0.987
1.307
1.636
1.659
1.672
1.695
1.722
1.746
1.813
1.851
1.955
1.974
1.994
2.104
2.139
2.208
2.342
2.354
2.392
2.414
2.434
3.739
3.775
3.986
4.010
4.042
4.084
4.130
4.178
4.211
4.244
4.283
4.307
4.401
4.428
4.510
4.533
5.091
5.162
6.701
10.884
2.003
2.263
13.539
3.778
0.493
2.657
0.431
0.500
0.909
0.891
0.802
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
23.301
26.189
26.281
26.391
28.668
28.768
29.361
29.411
29.464
29.620
29.694
30.145
33.365
38.742
60.584
62.417
62.875
63.044
65.503
65.860
76.435
77.362
77.608
77.875
78.707
140.029
140.062
145.004
145.069
169.148
169.284
169.538
169.691
169.741
169.976
171.009
171.076
199.711
199.743
1H NMR, 300 MHz, CDCl3
N
OAcAcO
AcO
O
ent-22
Ac
13C NMR, 101 MHz, CDCl3
S28
9 8 7 6 5 4 3 2 1 0 ppm
1.592
1.610
1.625
1.685
1.705
1.709
1.728
1.749
1.768
1.796
1.813
1.822
2.032
2.039
2.058
2.099
2.113
2.173
2.181
2.207
2.233
2.328
2.352
2.372
3.579
3.594
3.860
3.872
4.217
4.240
4.253
4.277
4.393
4.416
4.437
4.670
4.692
4.713
6.015
6.048
6.540
7.193
7.563
8.704
8.171
2.183
2.972
2.849
2.021
1.673
0.886
0.868
1.799
1.000
1.032
3.801
1.134
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
23.477
26.142
27.123
28.985
29.726
29.983
30.050
34.540
38.837
62.575
62.843
65.022
79.300
83.209
139.775
145.177
198.807
1H NMR, 300 MHz, Pyr.
13C NMR, 75 MHz, Pyr.
S29
9 8 7 6 5 4 3 2 1 0 ppm
1.321
1.339
1.355
1.368
2.017
2.031
3.650
3.660
3.664
3.674
3.818
3.831
3.843
3.857
4.189
4.198
4.208
4.454
4.478
4.503
4.527
4.546
4.564
4.570
4.588
4.618
4.642
4.699
4.722
4.912
4.916
4.932
4.934
4.936
4.963
4.967
4.997
5.001
5.791
5.804
7.256
7.265
7.270
7.275
7.277
7.281
7.287
7.291
7.294
7.297
7.300
7.309
7.314
7.316
7.318
7.327
7.328
7.332
5.336
2.066
1.065
2.089
1.000
1.010
1.016
1.032
0.991
1.018
1.014
1.044
1.222
0.852
1.078
1.035
0.964
0.971
0.793
0.969
14.835
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
26.914
28.986
29.418
30.724
33.891
67.392
68.779
71.253
72.871
73.615
73.758
83.507
114.336
127.803
127.825
127.890
127.971
127.981
128.019
128.480
128.519
128.531
138.186
138.332
138.384
139.280
1H NMR, 500 MHz, CDCl3
13C NMR, 126 MHz, CDCl3
H3
H3 H6
H4H6
S30
H5 H2 H4 6
H2 4 H5
(H6, H
COSY, 5
NOES
5)
500MHz, CDC
Y, 500MHz, C
Cl3
CDCl3
S31
9 8 7 6 5 4 3 2 1 0 ppm
1.081
1.103
1.232
1.243
1.257
1.284
1.315
1.925
1.950
1.984
2.005
3.568
3.592
3.685
3.713
3.777
3.791
3.807
4.120
4.141
4.155
4.219
4.242
4.261
4.271
4.381
4.521
4.561
4.586
4.613
4.645
4.679
4.721
4.753
4.793
4.836
4.912
4.942
4.945
5.000
5.049
5.064
5.106
5.137
5.179
5.196
5.237
5.733
5.756
5.767
5.790
7.196
7.214
7.228
7.239
7.297
7.320
7.352
3.164
4.449
0.760
2.729
0.611
0.405
1.994
1.035
2.011
1.300
3.765
1.150
2.165
1.907
0.974
20.000
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
66.956
127.770
127.988
136.624
138.923
26.377
26.492
28.717
28.860
28.923
31.861
33.225
33.697
57.816
57.915
62.329
62.653
66.891
69.100
70.563
71.919
72.263
72.491
72.727
73.035
78.027
79.877
81.064
114.440
127.378
127.437
127.526
127.690
128.035
128.257
128.416
128.543
136.768
138.321
138.387
138.771
139.031
154.555
154.690
1H NMR, 300 MHz, CDCl3
13C NMR, 75 MHz, CDCl3
S32
9 8 7 6 5 4 3 2 1 0 ppm
1.020
1.074
1.147
1.242
1.261
1.269
1.461
1.475
1.490
1.503
1.516
1.634
1.887
1.908
3.554
3.566
3.579
3.591
3.691
3.705
3.784
4.135
4.233
4.247
4.263
4.296
4.316
4.374
4.525
4.549
4.591
4.614
4.640
4.678
4.702
4.753
4.776
4.798
4.821
5.016
5.040
5.071
5.096
5.139
5.164
5.199
5.223
7.183
7.193
7.219
7.233
7.252
7.266
7.283
7.294
7.323
7.355
7.368
4.129
5.495
1.938
2.143
2.532
0.415
1.976
1.013
2.013
1.264
1.375
2.358
0.998
2.000
19.884
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
25.578
26.400
29.177
29.216
31.697
32.659
33.172
57.719
57.815
62.262
62.567
62.768
66.839
69.004
70.459
71.818
72.158
72.398
72.635
72.866
72.946
77.512
77.929
79.781
80.964
127.189
127.309
127.351
127.446
127.595
127.695
127.944
128.027
128.181
128.328
128.456
136.526
136.624
138.213
138.283
138.642
138.913
154.487
154.639
1H NMR, 500 MHz, CDCl3
13C NMR, 126 MHz, CDCl3
S33
9 8 7 6 5 4 3 2 1 0 ppm
1.060
1.096
1.110
1.228
1.241
1.262
1.334
1.348
1.360
1.730
1.765
1.780
1.794
3.315
3.329
3.343
3.579
3.595
3.759
3.766
3.778
3.787
3.810
4.118
4.127
4.131
4.139
4.254
4.271
4.283
4.372
4.516
4.540
4.554
4.613
4.640
4.678
4.750
4.773
5.070
5.095
5.140
5.165
7.168
7.182
7.191
7.204
7.220
7.236
7.248
7.263
7.282
7.295
7.319
7.335
7.349
4.016
4.299
1.310
0.657
1.956
0.713
1.846
0.602
0.384
1.969
1.001
1.927
1.294
1.390
2.383
0.998
2.000
19.641
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
28.806
28.970
29.605
31.603
32.534
32.578
32.990
33.825
33.876
57.640
57.719
62.111
62.458
66.685
66.748
68.895
70.351
71.739
72.068
72.299
72.531
72.748
72.820
77.364
77.862
79.669
80.845
127.073
127.186
127.235
127.329
127.482
127.499
127.571
127.798
127.832
127.954
128.066
128.226
128.349
136.440
136.565
138.132
138.190
138.571
138.837
154.295
154.461
1H NMR, 500 MHz, CDCl3
13C NMR, 300 MHz, CDCl3
S34
9 8 7 6 5 4 3 2 1 0 ppm
1.081
1.133
1.229
1.354
1.397
1.607
1.612
1.618
1.646
1.657
1.667
1.688
1.705
1.743
1.772
1.790
1.808
1.826
1.841
1.855
1.992
2.050
2.058
2.211
2.226
2.245
2.259
2.277
2.294
2.339
2.354
3.784
3.797
4.127
4.148
4.162
4.219
4.243
4.263
4.378
4.525
4.564
4.611
4.643
4.676
4.751
4.791
5.061
5.105
5.138
7.193
7.223
7.234
7.252
7.293
7.314
7.321
7.350
13.637
7.882
3.237
0.611
0.449
1.921
1.001
2.111
1.178
3.654
1.104
2.000
19.695
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
33.288
62.358
69.093
72.222
72.685
77.360
81.114
24.880
26.587
27.198
28.074
29.467
29.686
31.884
33.924
42.004
50.815
57.908
62.678
66.843
70.562
71.906
72.462
72.989
78.055
79.916
127.302
127.402
127.586
127.637
127.987
128.196
128.361
128.485
136.758
138.388
138.769
154.643
213.451
1H NMR, 300 MHz, CDCl3
13C NMR, 75 MHz, CDCl3
S35
7 6 5 4 3 2 1 0 ppm
1.536
1.714
1.746
1.772
1.784
1.815
1.834
1.874
1.889
2.053
2.059
2.074
2.093
2.103
2.111
2.132
2.144
2.151
2.164
2.176
2.365
2.382
2.401
2.421
3.332
3.338
3.343
3.348
3.353
3.358
3.416
3.435
3.446
3.464
3.475
3.493
3.753
3.890
3.930
3.951
3.968
3.986
4.004
4.015
4.202
4.915
14.402
6.553
1.855
2.278
1.103
1.000
2.955
0.948
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
25.785
27.527
28.201
29.291
30.332
30.608
30.765
31.991
35.335
42.798
51.808
59.427
62.014
63.397
71.771
77.454
216.303
1H NMR, 300 MHz, MeOD
13C NMR, 75 MHz, MeOD
S36
9 8 7 6 5 4 3 2 1 0 ppm
1.179
1.201
1.258
1.326
1.345
1.359
1.367
1.393
1.426
1.452
1.641
1.660
1.681
1.700
1.760
1.841
1.858
1.977
2.011
2.029
2.048
2.072
2.107
2.127
2.151
2.220
2.242
2.254
2.281
2.305
2.368
2.392
2.395
3.721
3.742
3.959
3.980
4.277
4.283
4.308
4.322
4.337
4.351
4.359
4.452
4.469
4.474
4.490
4.546
5.311
5.319
5.325
5.335
5.341
5.351
5.357
5.365
5.457
13.557
4.894
11.985
4.671
0.334
0.572
1.500
0.934
0.362
1.515
0.366
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
26.418
27.175
28.135
29.306
29.348
29.395
29.451
29.658
29.704
29.782
31.272
33.895
33.984
34.655
42.039
42.068
50.821
55.107
56.700
60.865
62.677
63.461
63.648
68.932
70.106
74.130
75.217
169.630
169.692
169.831
170.027
170.063
170.306
170.416
213.654
213.735
1H NMR, 500 MHz, CDCl3
13C NMR, 126 MHz, CDCl3
S37
9 8 7 6 5 4 3 2 1 0 ppm
1.165
1.179
1.189
1.201
1.276
1.300
1.324
1.342
1.356
1.452
1.777
1.948
1.960
1.976
1.992
2.006
2.034
2.053
2.077
2.111
2.132
2.156
2.224
2.344
2.354
2.401
2.418
2.433
3.718
3.723
3.745
3.962
3.982
4.276
4.285
4.304
4.313
4.340
4.358
4.368
4.431
4.453
4.471
4.480
4.499
4.546
4.551
5.313
5.325
5.342
5.362
5.373
5.442
5.453
5.463
6.686
6.696
6.707
12.030
1.739
2.668
5.192
6.419
1.883
1.873
1.782
0.364
0.617
1.596
0.983
0.380
1.602
0.380
0.861
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
23.105
23.279
26.164
26.322
26.452
28.575
28.663
29.320
29.369
29.543
29.604
31.303
34.675
38.722
55.133
56.726
60.890
62.700
63.483
63.674
68.954
70.129
74.150
75.235
139.970
140.009
145.009
145.089
169.659
169.712
169.866
170.053
170.088
170.329
170.450
199.733
1H NMR, 400 MHz, CDCl3
13C NMR, 101 MHz, CDCl3
S38
9 8 7 6 5 4 3 2 1 0 ppm
1.184
1.210
1.237
1.313
1.337
1.562
1.581
1.710
1.730
1.752
1.772
2.031
2.064
2.087
2.104
2.120
2.138
2.173
2.198
2.223
2.330
2.353
2.374
3.875
3.891
4.205
4.335
4.364
4.407
4.511
4.535
4.572
4.649
6.315
6.546
6.669
7.188
7.555
8.705
8.265
1.592
2.679
3.204
1.282
1.161
1.036
0.975
2.107
0.973
1.027
5.038
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
23.911
26.567
27.743
29.409
30.074
30.160
30.288
30.413
33.530
39.284
61.020
62.086
63.140
72.990
78.858
140.248
145.488
199.137
1H NMR, 300 MHz, Pyr.
13C NMR, 75 MHz, Pyr.
S39
9 8 7 6 5 4 3 2 1 0 ppm
1.354
1.581
1.610
1.752
1.913
1.925
1.965
1.986
2.008
2.105
2.144
2.167
2.188
2.397
2.419
2.442
3.227
3.243
3.299
3.328
3.366
3.479
3.498
3.668
3.762
3.801
3.825
3.844
3.858
4.144
4.945
6.846
6.859
6.872
14.324
1.702
1.478
2.292
1.000
1.292
3.465
1.121
0.536
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
24.392
27.167
27.903
30.011
30.481
30.523
30.711
33.760
39.593
60.869
62.145
62.615
72.787
78.454
140.839
148.204
202.159
13C NMR, 75 MHz, MeOD
13C NMR, 300 MHz, MeOD
S40
9 8 7 6 5 4 3 2 1 0 ppm
1.317
1.352
1.360
1.366
1.380
2.014
2.029
3.656
3.666
3.670
3.680
3.805
3.818
3.835
3.849
4.186
4.195
4.205
4.447
4.471
4.492
4.516
4.535
4.558
4.562
4.586
4.616
4.639
4.685
4.708
4.910
4.912
4.914
4.931
4.933
4.935
4.962
4.966
4.996
5.000
5.788
5.802
7.248
7.258
7.260
7.261
7.267
7.273
7.278
7.283
7.288
7.292
7.295
7.304
7.312
7.321
7.325
7.327
5.326
1.985
1.263
2.046
1.000
1.002
1.004
1.027
1.003
1.001
1.021
1.043
1.083
0.998
1.056
1.024
0.981
0.976
0.972
0.855
15.329
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
26.890
28.980
29.434
30.530
33.886
67.389
68.927
70.927
72.835
73.577
73.708
77.273
83.309
114.327
127.766
127.787
127.860
127.939
127.971
127.991
128.454
128.499
138.200
138.340
138.409
139.259
1H NMR, 500 MHz, CDCl3
13C NMR, 126 MHz, CDCl3
H3 H4 H6 H2 4 H5
(H6, H
H3 H
S41
H5)
H2 H4 H6 H5
NOESY,
COSY, 5
500MHz, CD
500MHz, CDC
DCl3
Cl3
S42
9 8 7 6 5 4 3 2 1 0 ppm
1.028
1.079
1.095
1.116
1.136
1.232
1.320
1.930
1.942
1.990
2.001
3.575
3.592
3.692
3.785
3.806
4.137
4.140
4.251
4.267
4.299
4.318
4.381
4.530
4.554
4.597
4.622
4.646
4.686
4.760
4.784
4.805
4.829
4.918
4.939
4.957
4.991
5.017
5.042
5.077
5.102
5.144
5.168
5.205
5.230
5.738
5.753
5.770
5.786
7.190
7.194
7.225
7.239
7.259
7.301
7.325
7.361
7.373
3.282
4.199
0.420
2.659
0.632
0.391
2.012
1.024
2.048
1.246
1.415
2.432
1.029
1.993
2.064
0.961
20.000
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
28.680
28.830
28.891
31.826
33.193
33.637
33.675
57.774
57.883
62.295
62.622
66.853
66.927
69.065
70.532
71.882
72.233
72.454
72.695
72.921
73.006
77.575
77.994
79.834
81.027
114.418
114.460
127.230
127.351
127.402
127.499
127.658
127.744
127.958
128.004
128.089
128.232
128.387
128.513
136.598
136.739
138.290
138.353
138.739
138.882
138.996
154.506
154.650
1H NMR, 500 MHz, CDCl3
13C NMR, 126 MHz, CDCl3
S43
9 8 7 6 5 4 3 2 1 0 ppm
1.020
1.075
1.147
1.243
1.260
1.284
1.417
1.461
1.476
1.490
1.503
1.517
1.530
1.887
1.909
3.553
3.565
3.578
3.591
3.783
3.789
3.806
4.138
4.145
4.234
4.247
4.264
4.279
4.374
4.525
4.549
4.615
4.641
4.679
4.753
4.777
4.799
5.041
5.072
5.097
5.140
5.165
5.200
7.171
7.186
7.191
7.208
7.219
7.232
7.251
7.266
7.282
7.294
7.301
7.314
7.324
7.356
7.370
10.181
1.957
0.420
1.672
2.535
0.451
1.991
1.017
2.030
1.266
1.382
2.373
0.994
2.000
20.155
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
26.425
29.197
29.239
31.724
32.690
33.200
57.746
57.842
62.288
62.590
62.828
66.863
66.901
69.031
70.486
71.845
72.189
72.427
72.663
72.897
72.976
77.363
77.539
77.957
79.814
80.998
127.217
127.335
127.378
127.473
127.616
127.636
127.723
127.946
127.972
128.051
128.155
128.210
128.354
128.482
136.558
136.656
138.240
138.312
138.672
138.940
154.512
154.663
1H NMR, 500 MHz, CDCl3
13C NMR, 126 MHz, CDCl3
S44
9 8 7 6 5 4 3 2 1 0 ppm
1.068
1.100
1.121
1.238
1.251
1.259
1.294
1.351
1.365
1.753
1.790
1.804
1.819
3.341
3.347
3.361
3.374
3.576
3.592
3.764
3.771
3.782
3.803
4.126
4.134
4.139
4.147
4.248
4.266
4.282
4.377
4.382
4.528
4.552
4.619
4.647
4.688
4.759
4.783
5.075
5.100
5.142
5.167
7.175
7.180
7.189
7.209
7.220
7.234
7.253
7.267
7.286
7.297
7.305
7.315
7.327
7.357
7.371
3.891
4.220
1.259
0.652
2.104
0.614
1.842
0.602
0.402
1.952
1.002
1.983
1.235
1.367
2.376
0.997
2.000
19.539
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
28.932
29.090
29.714
31.727
32.651
32.697
33.109
33.930
33.987
57.736
57.826
62.222
62.555
66.807
66.875
69.001
70.459
71.847
72.189
72.413
72.649
72.871
72.949
77.521
77.954
79.787
80.966
127.189
127.304
127.349
127.445
127.567
127.605
127.692
127.914
127.947
128.065
128.183
128.339
128.462
136.539
136.668
138.231
138.293
138.672
138.938
154.437
154.595
1H NMR, 500 MHz, CDCl3
13C NMR, 126 MHz, CDCl3
S45
9 8 7 6 5 4 3 2 1 0 ppm
1.083
1.152
1.231
1.346
1.389
1.597
1.608
1.635
1.657
1.678
1.694
1.779
1.798
1.816
1.831
1.844
1.980
2.043
2.050
2.201
2.220
2.238
2.245
2.264
2.284
2.331
2.346
3.785
3.797
4.126
4.147
4.161
4.245
4.265
4.274
4.376
4.522
4.561
4.611
4.642
4.673
4.748
4.788
5.059
5.104
5.137
7.192
7.208
7.217
7.229
7.246
7.262
7.289
7.300
7.306
7.319
13.482
7.588
3.222
0.645
0.453
1.911
0.991
2.058
1.155
3.633
1.124
2.000
20.221
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
33.232
62.296
69.047
72.643
77.372
81.041
24.832
26.534
27.148
28.025
29.419
29.637
31.839
33.880
41.953
50.751
57.860
62.637
66.787
70.501
71.862
72.180
72.414
72.932
78.005
79.858
127.258
127.354
127.541
127.591
127.938
128.148
128.314
128.438
136.713
138.338
138.719
154.583
213.350
1H NMR, 300 MHz, CDCl3
13C NMR, 75 MHz, CDCl3
S46
7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 ppm
1.398
1.553
1.601
1.615
1.635
1.651
1.692
1.726
1.750
1.763
1.790
1.855
1.867
1.884
1.900
1.914
2.033
2.053
2.071
2.090
2.112
2.126
2.145
2.158
2.343
2.360
2.377
2.398
3.233
3.249
3.308
3.312
3.320
3.325
3.330
3.335
3.519
3.832
3.850
4.148
4.898
4.916
14.866
6.362
1.806
2.402
1.000
0.994
2.975
0.986
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
25.763
27.721
28.197
29.261
30.420
30.586
30.788
33.554
35.287
42.773
51.765
60.676
62.036
62.501
72.613
78.245
216.156
1H NMR, 300 MHz, MeOD
13C NMR, 75 MHz, MeOD
S47
9 8 7 6 5 4 3 2 1 0 ppm
1.636
1.671
1.703
1.744
1.848
1.858
2.029
2.051
2.077
2.111
2.132
2.154
2.222
2.253
2.280
2.317
2.339
2.353
2.396
3.730
3.765
3.951
3.984
4.274
4.285
4.322
4.346
4.369
4.382
4.416
4.451
4.476
4.486
4.511
4.541
4.567
5.308
5.323
5.345
5.367
5.382
5.438
5.452
5.463
5.478
12.553
4.558
11.803
4.814
0.359
0.576
1.536
1.253
1.445
0.319
200 180 160 140 120 100 80 60 40 20 0 ppm
20.544
20.698
20.815
21.911
22.814
24.643
26.014
26.139
26.913
27.890
29.079
29.183
29.391
29.439
30.999
33.675
33.750
34.373
41.771
50.541
54.917
56.498
60.588
62.451
63.214
63.393
68.729
69.866
73.925
74.991
169.411
169.601
169.719
170.032
213.293
1H NMR, 300 MHz, CDCl3
13C NMR, 75 MHz, CDCl3
S48
9 8 7 6 5 4 3 2 1 0 ppm
1.159
1.173
1.184
1.196
1.255
1.269
1.293
1.317
1.335
1.349
1.420
1.446
1.471
1.628
1.635
1.953
1.969
1.985
2.000
2.028
2.046
2.071
2.105
2.126
2.150
2.218
2.338
2.348
2.394
2.411
2.427
3.712
3.717
3.739
3.955
3.976
4.269
4.279
4.307
4.334
4.352
4.362
4.425
4.447
4.466
4.474
4.493
4.540
5.307
5.319
5.336
5.356
5.367
5.436
5.447
5.456
6.691
6.700
12.492
1.673
2.582
5.196
6.490
1.959
1.805
1.741
0.359
0.613
1.575
0.970
0.391
1.595
0.381
0.825
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
23.088
23.270
26.151
26.303
26.434
28.562
28.650
29.306
29.352
29.530
29.590
31.289
34.663
38.709
55.132
56.721
60.875
62.691
63.471
63.660
68.945
70.119
74.142
75.221
139.957
140.001
145.000
145.082
169.645
169.702
169.864
170.039
170.077
170.325
170.441
199.722
1H NMR, 400 MHz, CDCl3
13C NMR, 101 MHz, CDCl3
S49
7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 ppm
1.538
1.569
1.598
1.687
1.705
1.722
1.734
1.943
1.963
1.984
2.006
2.027
2.104
2.143
2.154
2.167
2.188
2.371
2.395
2.404
2.419
2.440
3.151
3.179
3.196
3.222
3.322
3.327
3.332
3.366
3.437
3.447
3.641
3.663
3.733
3.758
3.773
3.779
3.794
3.806
3.824
3.845
3.864
3.882
4.115
4.127
4.139
6.841
6.855
6.868
14.496
1.570
1.568
2.846
0.951
0.965
3.295
1.000
0.666
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
30.605
24.248
27.018
27.812
29.875
30.358
30.388
30.584
33.981
39.443
61.016
62.003
62.306
72.819
78.493
140.698
148.021
201.979
1H NMR, 300 MHz, MeOD
13C NMR, 75 MHz, MeOD
S50
9 8 7 6 5 4 3 2 1 0 ppm
1.255
1.340
1.378
1.384
1.394
1.428
1.442
1.462
1.481
1.495
1.510
1.648
1.856
1.873
2.029
2.045
3.137
3.151
3.170
3.184
3.499
3.514
3.526
3.538
3.554
3.577
3.594
3.761
3.784
3.792
3.932
4.423
4.431
4.452
4.462
4.493
4.535
4.564
4.914
4.940
4.962
5.005
5.733
5.750
5.758
5.766
5.775
5.792
5.800
5.809
5.817
5.834
6.144
7.245
7.277
7.285
7.308
7.319
4.056
1.140
1.070
2.064
1.000
1.062
1.036
2.061
1.031
6.288
1.966
0.993
0.988
15.730
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
26.155
28.611
29.180
33.824
68.465
70.074
70.216
71.757
71.816
73.462
84.743
86.848
114.505
127.742
127.801
127.837
127.994
128.111
128.465
128.496
138.199
138.314
139.061
N
OH
BnO OBn
BnO
ent-16a 1H NMR, 400 MHz, CDCl3
13C NMR, 101 MHz, CDCl3
H3
H4 H
H3
H4 H6
S51
H5 H2
6
H5 H2
6
(
(H5, H3)
COSY, 500 M
NO
H5, H6)
MHz, CDCl3
ESY, 500MHz
z, CDCl3
S52
9 8 7 6 5 4 3 2 1 0 ppm
1.054
1.078
1.099
1.125
1.148
1.172
1.196
1.803
1.826
1.906
1.927
3.344
3.379
3.408
3.414
3.628
3.643
3.658
3.691
3.712
3.734
3.962
4.064
4.214
4.221
4.234
4.240
4.255
4.259
4.282
4.315
4.334
4.355
4.374
4.463
4.497
4.517
4.537
4.557
4.795
4.829
4.846
4.851
4.909
4.950
5.048
5.090
5.132
7.057
7.088
7.101
7.114
7.133
7.151
7.160
7.179
7.212
7.223
4.445
1.931
1.016
1.418
1.050
2.678
0.627
1.540
0.626
2.436
1.024
1.015
1.458
1.644
1.209
1.014
0.895
20.000
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
25.943
28.430
28.633
29.926
31.253
33.481
33.653
62.586
62.929
64.643
64.976
66.805
66.869
67.769
68.722
70.811
71.007
71.154
72.961
73.028
81.922
83.078
83.286
84.355
114.479
127.533
127.588
127.644
127.692
127.717
127.768
128.015
128.090
128.170
128.338
128.377
128.447
128.505
136.579
136.633
137.636
137.671
137.889
137.939
138.289
138.530
138.710
138.847
154.226
154.647
1H NMR, 300 MHz, CDCl3
13C NMR, 75 MHz, CDCl3
S53
9 8 7 6 5 4 3 2 1 0 ppm
1.140
1.179
1.259
1.311
1.424
1.446
1.470
1.501
1.514
1.682
1.701
3.439
3.472
3.505
3.518
3.540
3.563
3.570
3.592
3.614
3.705
3.719
3.737
3.749
3.791
3.808
3.859
4.048
4.119
4.132
4.159
4.346
4.356
4.371
4.389
4.418
4.438
4.459
4.479
4.567
4.600
4.608
4.618
4.640
4.658
5.011
5.052
5.142
5.184
5.226
7.194
7.205
7.231
7.261
7.286
7.303
7.320
7.333
6.688
4.669
0.538
1.018
0.936
0.958
2.590
0.486
1.533
0.502
2.484
0.994
1.075
1.494
0.986
1.000
19.854
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
25.525
25.565
26.428
26.531
28.995
29.071
29.953
31.441
32.710
62.686
62.923
63.029
64.773
65.011
66.932
67.898
68.844
70.954
71.124
71.262
73.074
73.135
82.075
83.219
83.503
84.544
127.605
127.665
127.713
127.757
127.789
127.843
128.096
128.144
128.238
128.418
128.529
128.586
136.705
137.766
137.999
138.379
138.627
154.345
154.758
1H NMR, 300 MHz, CDCl3
13C NMR, 75 MHz, CDCl3
S54
9 8 7 6 5 4 3 2 1 0 ppm
1.131
1.142
1.262
1.359
1.598
1.621
1.642
1.675
1.689
1.714
1.738
1.769
1.792
1.815
3.267
3.289
3.312
3.327
3.350
3.373
3.443
3.474
3.507
3.724
3.740
3.751
3.779
3.799
4.054
4.136
4.162
4.319
4.339
4.357
4.375
4.382
4.419
4.435
4.459
4.476
4.572
4.602
4.617
4.642
4.658
5.005
5.011
5.046
5.053
5.143
5.187
5.229
7.195
7.203
7.257
7.284
7.318
7.332
2.462
4.778
3.699
0.563
0.882
0.931
0.983
2.487
0.536
1.497
0.528
2.505
0.988
1.063
1.500
0.982
1.000
19.535
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
26.328
27.943
28.125
28.339
28.569
30.085
31.374
32.680
32.768
33.868
33.977
62.667
63.012
64.672
65.018
66.871
66.951
67.850
68.803
70.919
71.109
71.257
73.043
73.107
82.007
83.146
83.421
84.472
127.605
127.742
127.853
128.080
128.173
128.203
128.388
128.434
128.514
136.646
137.707
137.957
138.356
138.595
154.296
154.688
1H NMR, 300 MHz, CDCl3
13C NMR, 75 MHz, CDCl3
S55
9 8 7 6 5 4 3 2 1 0 ppm
1.157
1.255
1.335
1.344
1.611
1.620
1.640
1.648
1.667
1.675
1.697
1.712
1.736
1.832
2.001
2.018
2.040
2.223
2.242
2.264
2.303
2.360
3.469
3.734
3.746
3.795
3.817
4.153
4.316
4.354
4.371
4.396
4.418
4.444
4.459
4.485
4.566
4.599
4.606
4.618
4.639
4.658
5.012
5.053
5.145
5.185
5.225
7.173
7.179
7.195
7.215
7.220
7.241
7.263
7.286
7.307
7.323
7.335
10.837
5.950
2.630
3.071
1.000
2.542
0.492
1.521
0.514
4.571
1.498
0.990
0.991
19.873
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
27.204
28.152
29.270
29.491
29.719
29.786
30.186
31.525
33.963
42.079
50.833
62.662
62.984
64.784
65.094
66.876
67.879
68.826
70.921
71.082
71.233
73.040
73.107
82.066
83.198
83.403
84.488
127.592
127.654
127.704
127.782
128.112
128.242
128.413
128.450
128.511
128.579
136.718
137.752
137.986
138.376
138.622
154.310
154.757
213.618
213.712
1H NMR, 300 MHz, CDCl3
13C NMR, 75 MHz, CDCl3
S56
7 6 5 4 3 2 1 0 ppm
1.066
1.093
1.113
1.226
1.247
1.292
1.330
1.363
1.520
1.536
1.549
1.575
1.589
1.607
1.633
1.644
1.655
1.680
1.739
1.751
1.768
1.913
1.919
1.933
1.953
1.964
1.976
1.987
2.002
2.016
2.045
2.209
2.224
2.243
2.262
2.281
2.778
2.797
2.897
2.912
2.933
3.204
3.209
3.214
3.248
3.469
3.481
3.489
3.505
3.526
3.555
3.569
3.593
3.607
3.643
3.664
3.686
4.778
11.772
5.436
2.235
2.847
0.925
0.951
1.899
1.032
1.000
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
25.801
27.617
28.172
29.290
30.576
30.679
30.773
35.018
35.301
42.794
51.749
62.752
63.001
64.478
79.384
83.367
216.099
1H NMR, 300 MHz, MeOD
13C NMR, 75 MHz, MeOD
S57
9 8 7 6 5 4 3 2 1 0 ppm
1.236
1.259
1.292
1.306
1.354
1.365
1.399
1.446
1.632
1.662
1.701
1.718
1.741
1.757
1.765
1.781
1.804
1.823
1.837
1.851
1.866
2.044
2.087
2.097
2.105
2.113
2.136
2.204
2.227
2.246
2.265
2.281
2.296
2.300
2.311
2.360
3.728
3.736
3.766
4.007
4.042
4.074
4.081
4.090
4.212
4.244
4.272
4.282
4.390
4.400
4.422
4.497
4.509
4.530
4.541
5.086
5.099
5.160
10.721
5.229
12.753
4.382
0.539
1.415
1.142
0.419
0.510
0.947
0.906
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
24.957
26.192
26.286
27.117
28.106
29.273
29.412
29.624
30.076
33.307
33.892
33.944
34.011
42.033
42.084
50.785
60.500
62.343
62.800
62.983
65.415
65.763
76.363
77.804
78.636
169.068
169.220
169.456
169.625
169.870
170.916
170.988
213.499
213.631
1H NMR, 300 MHz, CDCl3
13C NMR, 75 MHz, CDCl3
S58
9 8 7 6 5 4 3 2 1 0 ppm
1.627
1.781
1.808
1.833
1.970
2.089
2.098
2.114
2.129
2.137
2.198
2.207
2.250
2.257
2.291
2.342
2.402
3.735
3.764
3.982
4.007
4.038
4.045
4.064
4.072
4.086
4.113
4.173
4.182
4.197
4.207
4.222
4.232
4.276
4.300
4.393
4.404
4.418
4.506
4.517
4.532
5.088
5.152
6.697
9.847
0.761
0.749
2.416
14.070
2.034
2.023
0.582
1.361
0.569
0.561
0.435
0.576
1.000
0.978
0.969
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
21.070
22.826
23.192
26.092
26.160
26.247
28.524
29.201
29.245
29.495
29.557
29.664
30.062
33.274
38.627
60.470
62.318
62.774
62.949
65.398
65.752
76.323
77.758
78.583
139.786
139.877
145.041
145.143
169.094
169.244
169.478
169.637
169.729
169.976
170.937
171.005
199.675
1H NMR, 400 MHz, CDCl3
13C NMR, 101 MHz, CDCl3
S59
7 6 5 4 3 2 1 0 ppm
1.670
1.696
1.802
1.830
1.850
1.868
1.922
1.932
1.948
1.955
1.968
1.989
2.011
2.032
2.155
2.178
2.375
2.399
2.409
2.423
2.444
2.575
2.600
2.624
2.637
3.214
3.236
3.260
3.283
3.322
3.327
3.332
3.366
3.386
3.478
3.744
3.767
3.785
3.805
3.817
3.827
3.855
3.869
3.921
3.940
3.959
4.905
6.856
6.868
6.882
9.107
2.311
1.526
1.020
1.115
1.863
0.434
1.063
1.000
3.364
1.207
0.484
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
24.207
26.998
27.130
29.726
30.124
30.171
30.486
32.589
39.417
60.296
63.784
65.169
77.213
80.983
140.569
148.167
202.021
1H NMR, 300 MHz, MeOD
13C NMR, 75 MHz, MeOD
S60
9 8 7 6 5 4 3 2 1 0 ppm
-0.000
1.464
1.482
1.487
1.493
1.495
1.500
1.558
2.062
2.075
3.199
3.534
3.569
3.588
3.601
3.789
3.794
3.799
3.804
3.926
3.931
3.934
3.939
4.450
4.463
4.474
4.477
4.543
4.554
4.567
4.932
4.934
4.936
4.952
4.954
4.956
4.977
4.981
5.011
5.015
5.091
5.785
5.798
7.258
7.266
7.277
7.284
7.286
7.294
7.300
7.305
7.307
7.309
7.314
7.319
7.328
7.334
7.337
3.177
1.020
2.079
1.000
1.010
1.006
2.019
1.008
6.249
0.994
1.001
0.766
0.980
14.873
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
25.947
28.452
33.975
68.342
69.885
70.197
71.703
71.786
73.425
84.648
86.837
114.702
127.712
127.770
127.802
127.951
128.089
128.433
128.453
128.467
138.167
138.205
138.291
138.768
1H NMR, 500 MHz, CDCl3
13C NMR, 126 MHz, CDCl3
H3 H4
H3
H4
H5H2
4 H6
H5H2
4 H6
(H5, H3
S61
COS
5
5
(H5, H6)
3)
SY, 500 MHz,
NOESY, 500M
CDCl3
MHz, CDCl3
S62
9 8 7 6 5 4 3 2 1 0 ppm
1.226
1.251
1.275
1.300
1.706
1.914
2.019
2.041
2.064
2.084
3.447
3.480
3.512
3.713
3.723
3.743
3.757
3.802
3.820
3.852
3.887
4.055
4.138
4.167
4.246
4.270
4.315
4.352
4.369
4.392
4.415
4.434
4.456
4.476
4.565
4.600
4.612
4.638
4.653
4.866
4.879
4.896
4.912
4.944
5.006
5.048
5.147
5.186
5.226
7.185
7.206
7.237
7.255
7.261
7.282
7.297
7.314
7.328
2.204
1.696
1.104
1.535
0.995
2.530
0.523
1.488
0.577
2.378
1.020
1.051
1.534
1.723
1.258
1.039
1.000
19.788
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
137.978
25.956
26.009
29.871
31.134
33.412
33.624
62.674
63.009
64.687
64.992
66.890
66.954
67.866
68.819
70.939
71.104
71.252
73.044
73.104
82.041
83.195
83.482
84.501
114.657
114.707
127.564
127.632
127.688
127.722
127.759
127.825
128.066
128.178
128.214
128.388
128.466
128.509
136.683
137.720
138.015
138.365
138.431
138.643
154.310
154.716
N
OBnBnO
BnO
ent-17b
Cbz
1H NMR, 300 MHz, CDCl3
13C NMR, 75 MHz, CDCl3
S63
9 8 7 6 5 4 3 2 1 0 ppm
3.450
3.470
3.482
3.494
3.559
3.571
3.584
3.715
3.721
3.732
3.740
3.747
3.770
3.784
3.801
3.834
3.856
4.051
4.130
4.135
4.156
4.240
4.260
4.326
4.335
4.351
4.366
4.386
4.411
4.423
4.447
4.469
4.573
4.599
4.604
4.625
4.648
5.014
5.019
5.039
5.044
5.145
5.170
5.188
5.213
7.169
7.183
7.195
7.207
7.225
7.246
7.262
7.285
7.294
7.309
7.319
7.336
7.344
2.464
3.015
2.092
1.302
0.523
1.900
1.047
1.909
0.547
0.461
1.467
0.493
2.339
2.122
1.462
0.980
1.000
19.516
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
25.439
26.281
26.307
30.073
31.369
32.525
32.650
62.658
62.802
62.849
62.991
64.666
64.931
66.919
66.976
67.837
68.785
70.921
71.099
71.244
73.050
73.115
81.979
83.130
83.458
84.468
127.602
127.653
127.671
127.722
127.777
127.846
127.859
128.100
128.227
128.409
128.449
128.515
128.579
136.652
137.692
137.721
137.948
138.001
138.338
138.585
154.345
154.731
1H NMR, 500 MHz, CDCl3
13C NMR, 126 MHz, CDCl3
S64
9 8 7 6 5 4 3 2 1 0 ppm
1.130
1.144
1.160
1.242
1.257
1.279
1.294
1.648
1.662
1.676
1.777
1.790
1.804
3.227
3.240
3.253
3.317
3.331
3.344
3.450
3.456
3.469
3.488
3.717
3.725
3.738
3.767
3.789
3.821
3.843
4.054
4.130
4.158
4.239
4.329
4.354
4.372
4.425
4.447
4.466
4.580
4.605
4.626
4.649
5.001
5.023
5.046
5.146
5.171
5.201
5.226
7.185
7.195
7.207
7.286
7.317
3.193
1.141
0.670
2.285
1.005
0.536
0.928
0.984
0.996
2.505
0.512
1.516
0.515
2.442
2.096
1.494
1.003
1.000
20.074
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
27.629
27.931
29.948
31.198
32.419
32.675
33.773
33.853
62.645
62.987
64.545
64.911
66.898
67.005
67.805
68.757
70.935
71.115
71.274
73.045
73.109
81.929
83.077
83.381
84.412
127.635
127.717
127.766
127.817
127.875
128.094
128.148
128.219
128.264
128.401
128.458
128.519
128.573
136.574
136.650
137.643
137.673
137.912
137.963
138.329
138.572
154.302
154.664
1H NMR, 500 MHz, CDCl3
13C NMR, 126 MHz, CDCl3
S65
9 8 7 6 5 4 3 2 1 0 ppm
1.150
1.222
1.246
1.256
1.268
1.599
1.639
1.657
1.685
1.695
1.735
1.759
1.801
1.817
1.834
2.001
2.028
2.192
2.210
2.230
2.250
2.268
2.288
2.336
2.350
2.394
3.469
3.502
3.747
3.794
3.815
4.154
4.351
4.364
4.392
4.415
4.442
4.456
4.482
4.565
4.597
4.615
4.637
4.655
5.011
5.051
5.144
5.185
5.226
7.192
7.213
7.228
7.260
7.283
7.303
7.319
8.090
4.375
1.612
3.093
2.214
1.342
0.926
2.391
0.495
1.484
0.543
4.445
1.528
0.964
1.000
19.619
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
24.897
26.510
27.093
28.073
29.335
29.564
30.140
33.878
42.018
50.693
62.683
63.026
64.756
65.091
66.841
68.841
70.922
71.074
71.214
73.020
73.080
82.098
83.199
83.444
84.504
127.555
127.600
127.660
127.778
128.029
128.177
128.355
128.392
128.472
128.521
136.708
137.739
137.983
138.367
138.609
154.273
154.703
213.409
N
OBnBnO
BnO
ent-19b
OCbz
1H NMR, 300 MHz, CDCl3
N
OBnBnO
BnO
ent-19b
OCbz
13C NMR, 75 MHz, CDCl3
S66
9 8 7 6 5 4 3 2 1 0 ppm
1.609
1.630
1.643
1.677
1.707
1.730
1.747
1.760
1.925
1.930
2.026
2.060
2.264
2.277
2.291
2.307
2.349
2.369
2.388
2.404
2.951
2.978
2.995
3.022
3.110
3.131
3.148
3.168
3.232
3.603
3.624
3.643
3.664
3.681
3.698
3.724
3.802
3.826
3.849
10.616
4.953
2.111
1.437
1.005
1.000
3.120
1.052
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
25.824
27.312
28.056
29.308
30.517
30.750
33.709
35.344
42.819
51.736
61.558
63.346
64.905
78.257
82.115
216.153
1H NMR, 300 MHz, D2O
13C NMR, 126 MHz, MeOD
S67
9 8 7 6 5 4 3 2 1 0 ppm
0.960
0.981
1.005
1.087
1.170
1.444
1.478
1.516
1.563
1.617
1.625
1.704
1.867
1.882
1.905
1.971
2.062
2.106
3.555
3.589
3.741
3.770
3.812
3.847
3.879
3.922
3.956
3.988
4.020
4.061
4.176
4.202
4.277
4.302
4.862
4.930
7.290
8.963
5.567
16.822
0.500
0.413
0.976
1.028
0.388
0.460
0.883
0.903
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
22.266
24.345
24.406
25.548
26.439
27.563
28.820
28.935
29.387
32.610
33.411
33.492
33.564
41.469
50.067
59.883
61.763
62.286
62.486
64.810
65.135
75.824
77.032
77.235
77.353
78.054
168.467
168.582
168.815
168.932
169.033
169.299
170.157
170.249
212.411
212.570
1H NMR, 300 MHz, CDCl3
13C NMR, 75 MHz, CDCl3
S68
9 8 7 6 5 4 3 2 1 0 ppm
1.720
1.746
1.765
1.780
1.812
1.849
1.884
1.975
1.997
2.086
2.090
2.101
2.112
2.133
2.202
2.344
2.384
2.392
2.410
2.428
3.734
3.767
3.975
4.008
4.044
4.076
4.125
4.175
4.207
4.240
4.269
4.277
4.306
4.393
4.420
4.503
4.514
4.536
4.547
5.083
5.093
5.161
6.698
6.707
6.713
4.177
14.455
3.932
0.524
0.505
1.000
1.059
0.437
0.519
0.978
0.966
0.918
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
21.041
22.835
23.209
26.007
26.054
26.106
28.448
28.581
29.148
29.479
29.513
29.681
30.002
33.215
38.618
60.523
62.349
62.846
63.008
65.422
65.760
76.395
77.583
77.803
78.665
139.754
139.858
145.011
145.151
169.042
169.168
169.437
169.569
169.664
169.896
170.872
170.941
199.470
199.539
1H NMR, 300 MHz, CDCl3
13C NMR, 75 MHz, CDCl3
S69
9 8 7 6 5 4 3 2 1 0 ppm
1.556
1.586
1.677
1.692
1.732
1.762
1.947
1.967
1.988
2.010
2.031
2.163
2.187
2.208
2.377
2.398
2.421
2.443
3.050
3.071
3.188
3.203
3.309
3.322
3.327
3.366
3.690
3.711
3.727
3.733
3.746
3.758
3.842
3.863
3.883
4.894
6.861
6.873
6.886
10.286
1.746
1.762
3.291
0.835
0.857
2.779
1.154
0.736
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 ppm
24.228
27.021
27.214
29.698
30.306
30.461
33.898
39.441
61.833
63.290
64.874
78.528
82.371
140.617
148.154
202.030
1H NMR, 300 MHz, MeOD
13C NMR, 75 MHz, MeOD
S70
4. Results of the bioassay
The enzymes α-glucosidase (from yeast; from rice), β-glucosidase (from almond; from bovine liver),
α-galactosidase (from coffee beans), β-galactosidase (from bovine liver), α-mannosidase (from jack bean),
β-mannosidase (from snail), α-L-fucosidase (from bovine kidney), α,α-trehalase (from porcine kidney),
amyloglucosidase (from A. niger), α-L-rhamnosidase (from Pdecumbens), β-glucuronidases (from bovine liver;
from E. coli), p-nitrophenyl glycosides, and various disaccharides were purchased from Sigma-Aldrich Co. Brush
border membranes were prepared from the rat small intestine according to the method of Kessler et al, (Kessler et
al 1978)1 and were assayed at pH 6.8 for rat intestinal maltase using maltose. ER α-glucosidase II was prepared
from rat liver according to the method of Hino et al.2 For rat intestinal glucosidases and porcine kitrehalase
activities, the reaction mixture (0.2 mL) contained 25 mM substrate and the appropriate amount of enzyme, and the
incubations were performed for 10 min at 37 ˚C. The reaction was stopped by heating at 100 ˚C for 3 min. After
centrifugation (600 g; 10 min), 0.035 mL of the resulting reaction mixture were added to 2.1 mL of the Glucose
CII-test Wako (Wako Pure Chemical Ind., Osaka, Japan). The absorbance at 505 nm was measured to determine
the amount of the released D-glucose. Other glycosidase activities were determined using an appropriate
p-nitrophenyl glycoside as substrate at the optimum pH of each enzyme. The reaction mixture (0.2 mL) contained 2
mM of the substrate and the appropriate amount of enzyme. The reaction was stopped by adding 0.4 mL of 400
mM Na2CO3. The released p-nitrophenol was measured spectrometrically at 400 nm. The assay results are
summarized in the following Table 3 and Table 4.
S71
Table 3. Concentrations of Broussonetine W and Its Analogues (4, ent-4, 3-epi-4, ent-3-epi-4, ent-4a, ent-4b)
Giving 50% Inhibition of Various Glycosidases.
IC50 (μM)
enzyme 4 ent-4 3-epi-4 ent-3-epi-4 ent-4a ent-4b
α-glucosidase
yeast 22 407 NIa (23.7%)b 310 758 622
rice 119 0.73 63 24 1.1 1.3
rat intestinal maltase 67 0.047 NI (43.9%) 3.5 0.19 0.2
intestinal isomaltase NI (29.3%) 1.5 NI (0%) 114 3.0 5.1
rat intestinal sucrase 216 0.20 111 3.4 0.25 0.28
ER α-glucosidase II NI (43.5%) 7.5 NI (13.2%) 200 15 21
β-glucosidase
almond 9.8 NI (39.5%) NI (31.8%) 92 NI (27.2%) NI (13.4%)
bovine liver 0.12 180 44 0.99 59 263
α-galactosidase
coffee bean NI (19.6%) NI (4.2%) NI (46.9%) NI (15.8%) NI (1.1%) NI (19.1%)
β-galactosidase
bovine liver 0.03 127 11 0.18 11 105
α-mannosidase
jack bean NI (15.1%) NI (0%) 385 NI (5.4%) NI (0%) NI (6.5%)
β-mannosidase
snail 282 NI (0%) NI (7.7%) NI (0%) NI (0%) NI (1.2%)
α-L-fucosidase
bovine kidney NI (16.8%) NI (0%) NI (12.3%) 20.4 NI (0%) NI (43.3%)
α, α-trehalase
porcine kidney NI (0.8%) NI (8.3%) NI (2.2%) NI (3.3%) NI (16.4%) NI (10.0%)
amyloglucosidase
Aspergilusniger 30 NI (0%) NI (2.8%) 448 NI (16.2%) NI (0%)
α-L-rhamnosidase
Penicillium decumbens NI (35.1%) 174 NI (47.3%) 166 591 1000
β-glucuronidase
E.coli 3.3 38 14 83.3 25 60
bovine liver NI (46.1%) 379 27 NI (40.2%) NI (46.7%) 186 a NI: No inhibition (less than 50% inhibition at 1000 μM). b ( ): inhibition % at 1000 μM.
S72
Table 4. Concentrations of Broussonetine W’s Analogues (20, ent-20, 3-epi-20, ent-3-epi-20, ent-20a, ent-20b)
Giving 50% Inhibition of Various Glycosidases
IC50 (μM)
enzyme 20 ent-20 3-epi-20 ent-3-epi-20 ent-20a ent-20b
α-glucosidase
yeast 80 NIa (32.7%)b NI (44.0%) NI (16.7%) NI (46.8%) NI (39.1%)
rice 70 1.7 7.9 33 1.4 3.7
rat intestinal maltase 32 0.15 1.6 13 0.11 0.14
intestinal isomaltase 663 3.7 40.3 366 2.1 3.8
rat intestinal sucrase 80 0.36 3.1 18 0.14 0.17
ER α-glucosidase II NI (46.9%) 14 157 458 12 25
β-glucosidase
almond 43 NI (37.2%) NI (25.4%) NI (35.5%) NI (35.8%) NI (39.0%)
bovine liver 0.10 14 27 40 237 372
α-galactosidase
coffee bean NI (13.8%) NI (14.6%) NI (29.4%) NI (13.5%) NI (0%) NI (0.7%)
β-galactosidase
bovine liver 0.046 8.5 35 14 42 129
α-mannosidase
jack bean NI (33.7%) NI (0%) 245 NI (2.4%) NI (1.2%) NI (0%)
β-mannosidase
snail 224 NI (15.2%) NI (0%) NI (0%) NI (0.9%) NI (0%)
α-L-fucosidase
bovine kidney NI (25.2%) NI (0.7%) NI (18.9%) 28 NI (35.7%) NI (7.6%)
α, α-trehalase
porcine kidney NI (0.8%) NI (3.3%) NI (0%) NI (0%) NI (10.0%) NI (13.2%)
amyloglucosidase
Aspergilusniger 41 NI (28.3%) NI (7.7%) NI (8.6%) NI (4.6%) NI (1.5%)
α-L-rhamnosidase
Penicillium decumbens NI (31.4%) 152 895 443 330 410
β-glucuronidase
E.coli 9.5 79 21 45 428 435
bovine liver NI (39.7%) NI (45.2%) NI (39.0%) NI (12.3%) NI (31.9%) 597 a NI: No inhibition (less than 50% inhibition at 1000 μM). b ( ): inhibition % at 1000 μM
(1) Kessler, M.; Acuto, O.; Strelli, C.; Murer, H.; Semenza, G. A. Biochem. Biophys. Acta. 1978, 506, 136.
(2) Hino, Y., Rothman, J. E. Biochemistry 1985, 24, 800-805.
