Detection, quantification and total synthesis of novel 3-hydroxykynurenine
glucoside-derived metabolites present in human lenses.
Nicholas A. Gada, Jasminka Mizdraka, David I. Pattisonb,c, Michael J. Daviesb,c, Roger J. W. Truscottd, Joanne F. Jamiea,*
5
10
a
Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, NSW 2109, Australia
b
The Heart Research Institute, Sydney, NSW 2042, Australia
c
Sydney Medical School, University of Sydney, Sydney, NSW 2006, Australia
d
Illawarra Health and Medical Research Institute, University of Wollongong, Wollongong, NSW 2522, Australia
*
Corresponding author. Tel: +61 2 9850 8283 Fax: +61 2 9850 8313
E-mail address: joanne.jamie@mq.edu.au (Joanne F. Jamie)
SUPPLEMENTARY DATA
15
Analytical data of 3OHKG-Y
3-Hydroxykynurenine O-β-D-glucoside yellow
(8-(β-D-glucopyranosyloxy)-1,2,3,4-tetrahydro-4-oxo-2-quinolinecarboxylic acid)
C16H19NO9 – (25.6 mg, 27%) as a pale yellow solid (~1:1 diastereomeric mixture). Calculated m/z M+H+ is 370.1133. Found m/z
M+H+ is 370.1132. ESI HRMS 370.1132 (M+H+, 11%), 392.0951 (M+Na+, 18%), 324.1076 (M+H+-HCOOH, 100%). ESI
20
HRMS/MS of M+H+, 370.1134 (100%), 208.0605 (M+H+-Glu, 93%); 1H NMR  (400 MHz, D2O) 7.49 (2x0.5H, d, J 8.1, H-5),
7.34 (2x0.5H, d, J 8.1, H-7), 6.78 (1H, dd, J 8.1, 8.1, H-6), 5.14 (0.5H, d, J 7.3, H-1’), 5.07 (0.5H, d, J 7.3, H-1’), 4.29 (1H, d, J
13.5, H-2), 3.96 (1H, d, J 12.5, H-6’), 3.80 (1H, dd, J 5.4, 12.5, H-6’), 3.69 (1H, m, H-2’), 3.67 (1H, m, H-3’), 3.63 (1H, m, H-5’),
3.55 (1H, m, H-4’), 3.01 (2x1H, m, H-3); 13C NMR  (CD3OD), 198.0 (CO-4), 172.0 (CO2H), 146.0 (C-8), 144.9 (C-8a), 144.7
(C-8a), 123.3 (C-7), 122.7 (C-7), 122.5 (C-5), 122.3 (C-5), 120.2 (C-4a), 118.8 (C-6), 102.9 (C-1’), 102.6 (C-1’), 77.6 (C-5’), 76.8
25
(C-3’), 74.1 (C-2’), 70.7 (C-4’), 61.8 (C-6’), 56.9 (C-2), 41.1 (C-3); max 228, 268 and 376, min 257 and 300 nm (Figure 2).
Analytical data of 3OHKG-W
4-(2-Amino-3-hydroxyphenyl)-2-hydroxy-4-oxobutanoic acid O-β-D-glucoside
C16H21NO10 – (10.1 mg, 10%) as a pale yellow solid (~1:1 diastereomeric mixture). Calculated m/z M+H+ is 388.1238. Found m/z
30
M+H+ is 388.1234. ESI HRMS/MS of M+H +, 226.0710 (M+H+-Glu, 100%), 208.0605 (M+H+-Glu – H2O, 22%); 1H NMR  (400
MHz, D2O), 7.65 (1H, d, J 8.1, ArH-6), 7.34 (1H, d, J 8.1, ArH-4), 6.79 (1H, dd, J 8.1, 8.1, ArH-5), 5.05 (1H, d, J 7.3, H-1’), 4.71
(1H, bd, J 5.4, H-2), 3.95 (1H, dd, J 2.0, 12.5, H-6’), 3.79 (1H, dd, J 5.2, 12.1, H-6’), 3.66 (1H, m, H-2’), 3.64 (1H, m, H-3’), 3.62
(1H, m, H-5’), 3.57 (1H, m, H-4’), 3.53 (2H, m, H-3); 13C NMR  (CD3OD), 201.3 (CO-4), 178.4 (C-1), 145.3 (ArC-3), 141.5
(ArC-2), 126.5 (ArC-6), 121.1 (ArC-4), 119.4 (ArC-1), 116.5 (ArC-5), 101.9 (C-1’), 76.6 (C-5’), 75.9 (C-3’), 73.3 (C-2’), 69.8
5
(C-4’), 67.6 (C-2), 60.9 (C-6’), 43.8 (C-3); max 227, 260 and 363, min 253 and 287 nm (Figure 2).
Analytical data of 3OHKG-D
(E)-4-(2-Amino-3-hydroxyphenyl)-4-oxo-but-2-enoic acid O-β-D-glucoside
C16H19NO9 – (3.2 mg, 3.4%) as a medium orange solid. Calculated m/z M+H+ is 370.1133. Found m/z M+H+ is 370.1133. ESI
10
HRMS/MS of M+H+, 370.1133 (M+H+, 100%), 208.0605 (M+H+-Glu, 63%); 1H NMR  (D2O) 7.86 (1H, d, J 15.6, H-3) 7.67
(1H, d, J 7.9, ArH-6), 7.37 (1H, d, J 7.9, ArH-4), 6.80 (1H, dd, J 8.0, 8.0, ArH-5), 6.76 (1H, d, J 15.8, H-2), 5.08 (1H, d, J 7.1, H1’), 3.96 (1H, dd, J 1.6, 12.5, H-6’), 3.80 (1H, dd, J 5.5, 13.1, H-6’), 3.68 (1H, m, H-2’), 3.65 (1H, m, H-3’), 3.62 (1H, m, H-5’),
3.56 (1H, m, H-4’); max 234, 282 and 409, min 257 and 340 nm (Figure 2).
15
Analytical data for AHAG
2-Amino-3-hydroxyacetophenone O-β-D-glucoside
C16H19NO9 – as a white solid. Calculated m/z M+H+ is 314.1234. Found m/z M+H+ is 314.1233. ESI HRMS/MS of M+H+,
152.0706 (M+H+-Glu, 100%), 278.1024 (12%); 1H NMR  (400 MHz, D2O), 7.66 (1H, d, J 8.1, ArH-6), 7.31 (1H, d, J 8.1, ArH4), 6.75 (1H, dd, J 8.1, 8.1, ArH-5), 5.03 (1H, d, J 7.3, H-1’), 3.92 (1H, dd, J 1.9, 12.5, H-6’), 3.76 (1H, dd, J 5.4, 12.5, H-6’), 3.63
20
(1H, m, H-2’), 3.62 (1H, m, H-3’), 3.58 (1H, m, H-5’), 3.51 (1H, m, H-4’), 2.60 (3H, m, H-1); 13C NMR  (CD3OD), 204.5 (CO),
145.5 (ArC-3), 139.5 (ArC-2), 127.3 (ArC-6), 121.0 (ArC-4), 120.3 (ArC-1), 117.3 (ArC-5), 101.8 (C-1’), 76.3 (C-5’), 75.6 (C3’), 73.0 (C-2’), 69.6 (C-4’), 60.6 (C-6’), 28.1 (CH3-1); max 226, 260 and 363, min 247 and 287 nm (Figure 2).