The Royal Society of Chemistry2-(((7-Methoxy-2-oxo-2H-chromen-4-yl)methyl)(methyl)amino)ethanoic...
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S1 ESI for ‘Fluorescent Molecular Logic Gates based on Photoinduced Electron Transfer (PET) Driven by a Combination of Atomic and Biomolecular Inputs’ by G. D. Wright, C. Y. Yao, T. S. Moody and A. P. de Silva Editor’s Note: Due to the 2020 global COVID-19 pandemic, not all characterization spectra could be made available prior to publication. This ESI replaces that originally published on 5 th May 2020. The characterisation data is now included. ChemComm Editorial Office 22 nd March 2021 S1. Synthesis procedures and characterization details for logic gates 1, 3, 5 and 7. Ethyl 2-(((7-methoxy-2-oxo-2H-chromen-4-yl)methyl)(methyl)amino)ethanoate, 1 To 4-(bromomethyl)-7-methoxycoumarin (1.0 g, 3.72 mmol) was added sarcosine ethyl ester hydrochloride (0.57 g, 3.74 mmol), K 2 CO 3 (3.0 g, anhydrous) and dichloromethane (15 ml) and the mixture stirred at approximately 70 o C for 18 hours. The mixture was then washed with water (3 x 50 ml), the organic layer dried with MgSO 4 and the solvent removed under vacuum, affording a yellow oil. Yield: 0.96 g (84 %) Electronic Supplementary Material (ESI) for Chemical Communications. This journal is © The Royal Society of Chemistry 2021
Transcript of The Royal Society of Chemistry2-(((7-Methoxy-2-oxo-2H-chromen-4-yl)methyl)(methyl)amino)ethanoic...
S1
ESI for ‘Fluorescent Molecular Logic Gates based on Photoinduced Electron Transfer (PET) Driven by a Combination of Atomic and Biomolecular Inputs’ by G. D. Wright, C. Y. Yao, T. S. Moody and A. P. de Silva
Editor’s Note: Due to the 2020 global COVID-19 pandemic, not all characterization spectra could be made available prior to publication. This ESI replaces that originally published on 5th May 2020. The characterisation data is now included.
ChemComm Editorial Office 22nd March 2021
S1. Synthesis procedures and characterization details for logic gates 1, 3, 5 and 7.
Ethyl 2-(((7-methoxy-2-oxo-2H-chromen-4-yl)methyl)(methyl)amino)ethanoate, 1To 4-(bromomethyl)-7-methoxycoumarin (1.0 g, 3.72 mmol) was added sarcosine ethyl ester hydrochloride (0.57 g, 3.74 mmol), K2CO3 (3.0 g, anhydrous) and dichloromethane (15 ml) and the mixture stirred at approximately 70 oC for 18 hours. The mixture was then washed with water (3 x 50 ml), the organic layer dried with MgSO4 and the solvent removed under vacuum, affording a yellow oil.
Yield: 0.96 g (84 %)
Electronic Supplementary Material (ESI) for Chemical Communications.This journal is © The Royal Society of Chemistry 2021
S2
Main characterization details are given in the paper.i.r. max (KBr): 3084, 2984, 2944, 2904, 2871, 2849, 2039, 1724, 1613, 1557, 1513, 1456, 1445, 1409, 1390, 1348, 1284, 1264, 1207, 1150, 1135, 1071, 1023, 995, 979, 964, 920, 886, 862, 849, 834, 814, 754, 742, 707, 650, 632, 576, 538, 486, 462 cm-1.
(R)-methyl 2-((7-methoxy-2-oxo-2H-chromen-4-yl)methylamino)propanoate, 3A procedure similar to that for 1 was employed, but with D-alanine methyl ester hydrochloride, affording a yellow oil.
Yield: 0.082 g (78 %)
S3
Main characterization details are given in the paper.i.r. max (KBr): 3080, 2952, 2843, 2361, 1724, 1615, 1558, 1513, 1456, 1394, 1348, 1283, 1265, 1209, 1150, 1074, 1023, 984, 851 cm-1.
(S)-methyl 2-((7-methoxy-2-oxo-2H-chromen-4-yl)methylamino)propanoate, 5
S4
A procedure similar to that for 1 was employed, but with L-alanine methyl ester hydrochloride, affording a yellow oil.
Yield: 0.073 g (69 %)
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9 (t)
6.43
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13'' (dd)
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12 (s)
3.87
19 (s)
3.77
21 (d)
1.37
14 (s)
1.59
2.92
3.78
1.21
3.00
2.99
1.05
1.02
2.07
1.00
1.37
1.38
1.59
3.41
3.42
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3.46
3.71
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3.75
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3.77
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4.02
4.05
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6.43
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6.83
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6.85
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6.86
7.26
7.57
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1,3
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613'' 14
13'
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2
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O7
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CH312
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O15
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O18
CH319
O20
CH321
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52.15
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56.45
77.16 CDCl3
101.16
110.76
112.23
112.50
125.20
153.47
155.76
161.60
162.75
176.00
Main characterization details are given in the paper.
S5
i.r. max (KBr): 3082, 2952, 2843, 2361, 1724, 1615, 1558, 1514, 1456, 1394, 1349, 1283, 1265, 1209, 1150, 1078, 1023, 985, 850 cm-1.
Ethyl 2-(methyl((10-((2,3,5,6,8,9,11,12-octahydrobenzo[b][1,4,7,10,13]pentaoxacyclopentadecin-15-yl)methyl)anthracen-9-yl)methyl)amino)ethanoate, 715-{[10-(Bromomethyl)-9-anthryl]methyl}-2,3,5,6,8,9,11,12-octahydro-1,4,7,10,13-benzopentaoxacyclopentadecineS1 (0.23 g, 0.415 mmol), sarcosine ethyl ester hydrochloride (0.0635 g, 0.415 mmol) and potassium carbonate (3 g, anhydrous) were refluxed in CH2Cl2 (10 ml) for 24 hours. Potassium carbonate was then filtered off and washed with CH2Cl2. The solvent was evaporated off producing a yellow oil.
Yield: 0.22 g (89 %)
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27,34 (d)8.60
30,31 (d)8.23
28,29,32,33 (m)7.48
17 (d)6.53
19 (s)6.72
16 (d)6.67
20 (s)4.94
35 (s)4.74
42 (q)4.20
38 (s)3.43
37 (s)2.51
43 (t)1.29
6,7,8,9,10,11,14,15 (m)3.80
2.9
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3
1.9
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16.0
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6
1.9
1
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7
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80.9
70.9
5
3.9
6
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1.2
71.2
91.3
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2.5
1
3.4
33.7
23.8
03.8
13.8
23.8
43.8
53.8
63.9
53.9
63.9
74.0
34.0
44.0
54.1
74.1
94.2
14.2
2
4.7
4
4.9
4
6.5
26.5
46.6
66.6
86.7
2
7.4
37.4
47.4
77.5
07.5
27.5
4
8.2
18.2
4
8.5
98.6
1
37
43
38
35
20
42
30,3127,346,7,8,9,10,11,14,1528,29,32,33
1916
17
O1
O2
O3
O4O
5
6
7
8
9
1011
12
13
1415
16
17
18
1920
21
22
23
24
25
26
27
28
29
3031
32
33
34
35
N36
CH337
38
39
O40
O41
42
CH343
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14.45
33.59
42.01
52.00
57.56
60.48
69.25
69.46
69.86
70.79
71.23
77.16 CDCl3
113.85
114.67
120.99
125.53
125.76
129.55
130.45
131.57
132.92
134.37
147.62
171.55
Main characterization details are given in the paper.i.r. max (KBr): 3436, 3065, 2918, 2865, 2361, 1729, 1671, 1608, 1587, 1559, 1514, 1447, 1424, 1358, 1272, 1253, 1234, 1187, 1138, 1071, 1051, 970, 940, 854, 808, 787, 760, 734, 663, 602, 576 cm-1.
S2. Synthesis procedures and characterization details for hydrolysis products 2, 6 and 8, and intermediates 9 and 10
tert-Butyl 2-(((7-methoxy-2-oxo-2H-chromen-4-yl)methyl)(methyl)amino)ethanoate, 9A procedure similar to that for 1 was employed, but with sarcosine tert-butyl ester hydrochloride, affording a yellow oil.
Yield: 1.13 g (91 %)
Found: C18H24NO5 [M+H+] 334.1654Required: C18H24NO5 [M+H+] 334.0535
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6 (d)
7.90
1 (dd)
6.85
3 (d)
6.82
9 (s)
6.35
12 (s)
3.87
16 (d)
3.82
13 (s)
3.27
17 (s)
2.42
21,23,24 (s)
1.49
9.56
3.10
2.06
2.11
3.18
0.95
0.94
1.03
1.00
1.49
2.42
3.27
3.82
3.83
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7.26 CDCl3
7.89
7.91 21,23,24
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O7
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CH317
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O19
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CH321
O22
CH323
CH324
1H NMR (CDCl3): δ1.49 (s, 9H, C(CH3)3), 2.42 (s, 3H, NCH3), 3.28 (s, 2H, ArCH2N), 3.83 (s, 2H, NCH2CO), 3.87 (s, 3H, OCH3), 6.35 (s, 1H, CHCO), 6.82 (d, 1H, ArH, J=3 Hz), 6.86 (dd, 1H, ArH, J=3, 9 Hz), 7.91 (d, 1H, ArH, J=9 Hz).
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81.55
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162.75
170.15
13C NMR (CDCl3): 28.2, 30.9, 42.2, 55.7, 100.8, 112.3, 126.3, 155.7, 161.5, 162.6, 169.9, 206.9.
S8
i.r. max (KBr): 2980, 2361, 1724, 1616, 1558, 1513, 1457, 1395, 1348, 1292, 1209, 1149, 1069, 1026, 840 cm-1.
2-(((7-Methoxy-2-oxo-2H-chromen-4-yl)methyl)(methyl)amino)ethanoic acid, 2To 9 (0.604 g, 1.81 mmol) was added formic acid (15 ml) in a 50 ml round-bottom flask and the solution heated at 60 oC for 6 hours while stirring. The solution was then allowed to cool and then placed in the fridge for 72 hours. The solvent was then removed on the rotary evaporator and approximately 5 ml H2O was added, followed by several drops of aqueous NH3 (0.2 M), causing a precipitate to form. The round-bottom flask was again placed in the fridge, for 24 hours. The solution was then filtered using a glass-sintered funnel and filter paper, affording an off-white solid.
Yield: 0.379 g (76 %)
Found: C14H16NO5 [M+H+] 278.1033Required: C14H16NO5 [M+H+] 278.1028
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6 (d)8.12
1 (dd)6.94
3 (d)6.89
9 (s)6.43
16 (s)3.92
12 (s)3.89
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13 (s)3.28
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3.1
41.8
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93.9
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1H NMR (CDCl3): δ2.47 (s, 3H, NCH3), 3.43 (s, 2H, ArCH2N), 3.80 (s, 3H, OCH3), 4.03 (s, 2H, NCH2CO), 6.34 (s, 1H, CHCO), 6.84 (d, 1H, ArH, J=3 Hz), 6.87 (dd, 1H, ArH, J=9, 3 Hz), 7.99 (d, 1H, ArH, J=9 Hz).
S9
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D3O
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1156.8
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8164.4
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13C NMR (CDCl3): δ28.6, 42.6, 56.1, 57.7, 59.1, 82.0, 101.2, 112.7, 112.8, 126.7, 152.7, 156.1, 161.9, 163.0, 170.2,
i.r. max (KBr): 3032, 3016, 2952, 2852, 1723, 1619, 1560, 1521, 1475, 1456, 1404, 1384, 1349, 1303, 1283, 1213, 1182, 1156, 1122, 1070, 1028, 987, 967, 950, 904, 879, 862, 848.
Hydrolysis product 4 could not be prepared conveniently since D-alanine tert-butyl ester hydrochloride was not commercially available at a reasonable price at the time.
(S)-tert-butyl 2-((7-methoxy-2-oxo-2H-chromen-4-yl)methylamino)propanoate, 10A procedure similar to that for 1 was employed, but with L-alanine tert-butyl ester hydrochloride, affording a yellow solid.
Yield: 0.607 g (97 %)
Found: C18H24NO5 [M+H+] 334.1667Required: C18H24NO5 [M+H+] 334.1668
S10
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6 (d)
7.60
1,3 (m)
6.84
9 (t)
6.42
13 (dd)
4.02
13 (dd)
3.73
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3.87
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3.28
20,23,24 (s)
1.49
22 (d)
1.32
3.08
9.56
0.98
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3.70
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6.84
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7.26 CDCl3
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1420,23,24
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1H NMR (CDCl3): 1.46 (d, 3H, C(CH3)H, J=7 Hz), 1.51 (s, 9H, C(CH3)3), 3.28 (q, 1H, C(CH3)H, J=7 Hz), 3.87 (dd, 2H, CH2NH, J=16, 88 Hz), 3.87 (s, 3H, OCH3), 3.89 (s, 1H, NH), 6.38 (s, 1H, CHCO), 6.82 (m, 1H, ArH), 6.89 (m, 1H, ArH), 7.62 (d, 1H, ArH, J=9 Hz).
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81.59
101.15
110.86
112.49
125.26
153.66
155.77
161.63
162.71
174.95
190.36
13C NMR (CDCl3): 16.9, 28.4, 48.1, 56.2, 57.4, 101.4, 111.1, 112.7, 125.5, 153.9, 155.9, 161.9, 163.0, 175.1.
S11
i.r. max (KBr): 2978, 2935, 1726, 1615, 1558, 1514, 1456, 1394, 1369, 1350, 1291, 1264, 1211, 1152, 1058, 1023, 990, 849 cm-1.
(S)-2-((7-methoxy-2-oxo-2H-chromen-4-yl)methylamino)propanoic acid, 6To 10 (0.2713 g, 0.814 mmol) was added formic acid (6 ml) and the solution heated at 60 oC while stirring for 6 hours. The solvent was then removed and precipitation encouraged by the addition of aqueous NH3 (0.2 M), affording an off-white solid.
Yield: 0.0964 g (43 %)
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7.81
1,3 (m)
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6.43
13' (m)
4.53
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4.27
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3.91
20 (d)
1.69
2.76
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1.70
3.91
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4.30
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O15
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OH18
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CH320
1H NMR (CDCl3): 1.58 (d, 3H, C(CH3)H, J=7 Hz), 3.88 (s, 3H, OCH3), 3.89 (s, 2H, CH2NH), 4.40 (q, 1H, C(CH3)H, J=7 Hz), 6.43 (s, 1H, ArH), 6.87 (m, 1H, ArH), 6.91 (m, 1H, ArH), 7.53 (m, 1H, ArH).
S12
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MSO
-d6
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9112.0
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8162.1
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8
13C NMR (CDCl3): 16.9, 53.9, 56.3, 101.5, 110.2, 111.5, 112.9, 113.1, 125.2, 150.8, 155.7, 161.5, 163.4, 174.1.
i.r. max (KBr): 3138, 1725, 1604, 1400, 1349, 1290, 1212, 1139, 1075, 1026, 1005, 984, 851, 763, 702 cm-1.
2-(Methyl((10-((2,3,5,6,8,9,11,12-octahydrobenzo[b][1,4,7,10,13]pentaoxacyclopentadecin-15-yl)methyl)anthracen-9-yl)methyl)amino)ethanoic acid, 8
7 (0.176 g, 0.3 mmol) was dissolved in THF (10 ml). A solution of NaOH (0.24 g, 6 mmol) in MeOH:H2O (1:1, v/v) was added. The solution was left to stir overnight. The solvent was then removed by evaporation. The yellow solid was dissolved in water and neutralised with acetic acid producing a yellow precipitate, which was filtered off.
Yield: 0.109 g (65 %)
Found: C33H37NO7Na [M+Na+] 582.2486Required: C33H37NO7Na [M+Na+] 582.2468
S13
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9000
10000CYY-CC9-001.10.fidPROTON.qub CD3OD_SPE {C:\Bruker\TopSpin3.2} desilva 13
30,31 (d)8.21
27,34 (d)8.79
28,29,32,33 (m)7.46
16,19 (m)6.79
17 (d)6.62
20 (s)4.93
35 (s)4.77
6,7,8,9,10,11,14,15 (m)3.74
38 (s)3.31
37 (s)2.33
2.8
9
1.7
7
16.0
9
1.8
9
2.1
6
0.9
6
1.9
6
4.0
7
2.0
0
1.9
4
2.3
3
3.3
1
3.6
33.6
43.6
53.6
83.6
93.7
03.7
13.7
53.7
63.7
73.7
73.8
73.8
83.8
83.8
94.0
34.0
34.0
44.0
54.7
74.8
7 C
D3O
D4.9
3
6.6
16.6
36.7
86.7
96.8
1
7.4
07.4
17.4
37.4
97.5
17.5
3
8.2
08.2
3
8.7
88.8
0
6,7,8,9,10,11,14,15
37
2038
3516,1930,3127,34
28,29,32,33
17
O1
O2
O3
O4O
5
6
7
8
9
1011
12
13
1415
16
17
18
1920
21
22
23
24
25
26
27
28
29
3031
32
33
34
35
N36
CH337
38
39
OH40
O41
1H NMR (MeOD-d4): δ2.75 (s, 3H, NCH3), 3.5-3.9 (m, 16H, CH2O), 3.65 (s, 2H, NCH2), 4.78 (s, 2H, AnthCH2N), 4.91 (s, 2H, AnthCH2Ar), 6.35 (d, 1H, ArH, J=8 Hz), 6.55 (d, 1H, ArH, J=8 Hz), 6.75 (s, 1H, ArH), 7.49 (dt, 4H, AnthH, J=7 Hz), 7.64 (dt, 2H, AnthH, J=7 Hz), 8.3 (d, 2H, AnthH, J=9 Hz).
3035404550556065707580859095100105110115120125130135140145150155
f1 (ppm)
-200
-100
0
100
200
300
400
500
600
700
800
900
1000
1100
1200
1300
1400
1500
1600
1700
1800
1900
2000
2100CYY-CC9-001.11.fidC13CPD_1024.QUB CD3OD_SPE {C:\Bruker\TopSpin3.2} desilva 13
33.8
6
42.4
4
53.1
5
62.6
6
68.7
869.1
069.2
469.3
869.5
869.7
9
115.2
6115.3
9
122.8
6
126.1
8126.5
2127.3
7
131.4
6132.9
2134.0
6
136.8
8
147.2
4
149.0
4
13C NMR (MeOD-d4): 38.7, 46.1, 48.7, 55.7, 58.0, 65.7, 74.2, 74.9, 75.7, 76.0, 119.7, 119.9, 125.8, 126.2, 135.9, 137.5, 139.3, 142.8, 153.0, 154.6, 175.9.
S14
i.r. max (KBr): 3035, 2867, 2361, 1633, 1511, 1451, 1424, 1360, 1271, 1252, 1137, 1055, 978, 938, 850, 763, 735, 692, 668, 603, 540 cm-1.
S3. Conditions and analytical procedures of enzyme screen1 (91.0 mg, 0.30 mmol) was suspended in methyl tert-butyl ether (8 ml) and 200 l of this suspension was added to 1 ml phosphate buffer (0.1 M) along with a 10 mg of enzyme in a small vial. The vials were then shaken overnight at room temperature. The samples were then put in the fridge for 4 nights. 700 l acetonitrile was then added to each vial, and the contents centrifuged, causing the protein to separate from the liquid. The mother liquor (150 l) was then removed, placed into an HPLC vial, which was then filled with MeCN (approx. 1 ml). This was then analysed by reversed phase HPLC carried out on a C18 column. A mobile phase with 5 % to 95 % solvent B in solvent A was run over 13 minutes, in which solvent A consisted of 0.1 % trifluoroacetic acid (TFA) in Milli Q® water and solvent B consisted of 0.1 % TFA in acetonitrile.
After having run the enzyme screen, the absorbance of the samples at 326 nm was reduced to 0.1 by dilution with pH 7 phosphate buffer, and their total fluorescence emission measured. The expected result was that those samples that showed a high conversion from the ester to the acid would be more fluorescent than those that did not. There is a linear relationship between the HPLC yield of 2 and the intensity of fluorescence. So the fluorescence method could be used to examine the activity of a particular enzyme.
S4. Time-dependent fluorescence during enzymatic hydrolysis As 1 was hydrolysed to 2 by CALB, the fluorescence (exc = 326 nm and em = 415 nm) increased gradually. The example below shows the effect of varying the CALB concentration.
S5. Standards for fluorescence quantum yieldsThe reference compounds used were (anthracen-9-ylmethyl)diethylamineS2 for the anthracene compounds 7-8 and 4-methyl-7-methoxycoumarinS3 for the coumarin compounds 1-6.
ReferencesS1. A. P. de Silva, H. Q. N. Gunaratne and C. P. McCoy, J. Am. Chem. Soc. 1997, 119, 7891. S2. R. A. Bissell, E. Calle, A. P. de Silva, S. A. de Silva, H. Q. N. Gunaratne, J. L. Habib-Jiwan, S. L. A. Peiris, R. A. D. D. Rupasinghe, T. K. S. D. Samarasinghe, K. R. A. S. Sandanayake and J. -P. Soumillion, J. Chem. Soc. Perkin Trans. 2 1992, 1559.S3. A. P. de Silva, H. Q. N. Gunaratne, P. L. M. Lynch, A. L. Patty and G. L. Spence, J. Chem. Soc. Perkin Trans. 2 1993, 1611.
0
100
200
300
400
500
600
0 100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700
1.4E-06
2.9E-06
4.3E-06
5.7E-06
7.2E-06
2.1E-05
IF
Time (s)
![Accumulation of glycerophosphocholine (GPC) renalcells ... · [methyl-14C]choline (3.6,uCi/ml,65,tM;1 Ci =37GBq).The radioactive mediumwasdecanted, andthenthe monolayers were rinsed](https://static.fdocuments.us/doc/165x107/5e54a4b0b6d2d248e21fa5e6/accumulation-of-glycerophosphocholine-gpc-renalcells-methyl-14ccholine-36uciml65tm1.jpg)
