Parthasarathi Dastidar, Siddhartha Sankar Jana et al.
J=2.4, 2.4 Hz, 1H), 4.52–4.45 (dd, J=3, 8.1 Hz, 1H), 3.85–3.80 (m, 1H),
3.79 (s, 3H), 3.78–3.55 (m, 4H), 2.43–2.27 (m, 1H), 2.23 (s, 3H), 2.20–
2.04 ppm (m, 1H); 13C NMR (300 MHz, CD3OD): d=174.8, 170.4, 168.5,
156.0, 138.6, 135.3, 134.2, 130.9, 130.8, 130.7, 128.7, 114.3, 113.1, 111.2,
101.2, 69.6, 55.1, 54.7, 37.3, 29.8, 12.3 ppm (Figure S3 in the Supporting
Information); FTIR (KBr pellet): n˜ =3367 (s, O-H stretch), 1712 (w, acid
C=O stretch), 1683 (s, amide C=O stretch), 1624 (s), 1473 (m), 1456 (m),
1319 (s), 1224 (m), 833 (w), 754 cmÀ1 (m); HRMS (ESI (CH3OH)): m/z
calcd for C24H23ClN2O6: 493.1142 [M+Na]+; found: 493.1142; elemental
analysis calcd (%) for C24H23ClN2O6·H2O·CH3OH: C 57.64, H 5.61, N
5.38; found: C 57.06, H 5.12, N 5.28.
168.7, 168.4, 156.1, 138.1, 135.5, 134.9, 131.7, 130.8, 129.6, 115.0, 114.6,
111.9, 102.8, 102.5, 100.1, 69.4, 67.7, 58.9, 56.0, 48.3, 38.3, 30.7, 17.8,
14.0 ppm (Figure S5 in the Supporting Information); FTIR (KBr pellet):
n˜ =3406 (s, O-H stretch), 3309 (s, amide N-H stretch) 1716 (w, acid C=O
stretch), 1656 (s, amide C=O stretch), 1637 (s), 1618 (m), 1542 (w), 1477
(m), 1454 (m), 1436 (m), 1400 (w), 1371 (m), 1321 (w), 1222 (m), 1149
(w), 1087 (m), 1035 (w), 831 (w), 796 (w), 754 cmÀ1 (m); HRMS (ESI
(CH3OH)): m/z calcd for C27H28ClN3O7: 564.1513 [M+Na]+; found:
564.1514;
elemental
analysis
calcd
(%)
for
C27H28ClN3O7·CH2Cl2·CH3OH: C 52.86, H 5.20, N 6.38; found: C 52.32,
H 4.62, N 6.59.
IND.SER
IND.HYP.THR
Yield: 74%; m.p. 164–1658C; 1H NMR (300 MHz, CD3OD, 258C): d=
7.65–7.62 (d, J=8.4 Hz, 2H), 7.51–7.48 (d, J=8.4 Hz, 2H), 7.05–7.04 (d,
J=2.1 Hz, 1H), 6.95–6.92 (d, J=9 Hz, 1H), 6.64–6.60 (dd, J=2.1, 2.1 Hz,
1H), 4.48 (s, 1H), 3.90–3.83 (dd, J=4.5, 3.6 Hz, 2H), 3.78 (s, 3H), 3.68 (s,
2H), 2.25 ppm (s, 3H); 13C NMR (300 MHz, CD3OD): d=171.74, 171.69,
168.4, 156.1, 138.6, 135.5, 134.1, 130.8, 130.8, 130.6, 128.7, 114.4, 113.3,
111.5, 100.8, 61.5, 55.0, 54.7, 30.6, 12.2 ppm (Figure S8 in the Supporting
Yield: 66%; m.p. 162–1638C; 1H NMR (300 MHz, CD3OD, 258C): d=
7.71–7.70 (d, J=3.6 Hz, 1H), 7.69–7.68 (d, J=3.6 Hz, 1H), 7.55–7.52 (d,
J=6.6 Hz, 2H), 7.00–6.90 (m, 2H), 6.65–6.62 (dd, J=1.8, 1.8 Hz, 1H),
4.66–4.62 (t, J=6 Hz, 1H), 4.53 (s, 1H), 4.35 (s, 1H), 3.87–3.82 (m, 2H),
3.80 (s, 3H), 3.78–3.64 (m, 3H), 3.34 (s, 2H), 2.23 (s, 3H), 2.22–2.10 (m,
1H; O-H proton), 1.16–1.15 ppm (d, J=4.5 Hz, 3H); 13C NMR
(300 MHz, CD3OD): d=174.9, 172.5, 171.8, 170.0, 157.7, 140.1, 136.9,
135.9, 132.4, 130.2, 115.9, 114.5, 112.8, 102.8, 71.1, 69.4, 60.7, 59.7, 56.9,
56.2, 39.0, 31.4, 20.3, 13.8 ppm (Figure S6 in the Supporting Information);
À
Information); FTIR (KBr pellet): n˜ =3307 (s, O H stretch), 1735 (m,
acid C=O stretch), 1676 (s, amide C=O stretch), 1647 (s), 1604 (m), 1596
(m), 1541 (m), 1477 (s), 1363 (m), 1317 (m), 1222 cmÀ1 (m); HRMS (ESI
(CH3OH)): m/z calcd for C22H21ClN2O6: 467.0986 [M+Na]+; found:
À
FTIR (KBr pellet): n˜ =3384 (s, O H stretch), 1718 (w, acid C=O stretch),
1677 (s, amide C=O stretch), 1637 (s), 1627 (s), 1477 (m), 1456 (m), 1400
(w), 1321 (m), 1222 (m), 1087 (m), 1014 (w), 754 cmÀ1; HRMS (ESI
(CH3OH)): m/z calcd for C28H30ClN3O8: 594.1619 [M+Na]+; found:
594.1619; elemental analysis calcd for C28H30ClN3O8·CH3OH: C 57.66, H
5.67, N 6.96; found: C 57.32, H 5.62, N 6.58.
467.0986; elemental analysis calcd (%) for C22H21ClN2O6·CH2Cl2:
C
52.14, H 4.38, N 5.29; found: C 52.22, H 3.87, N 5.88.
IND.THR
Yield: 78%; m.p. 160–1618C; 1H NMR (300 MHz, CD3OD, 258C): d=
7.65–7.62 (d, J=8.4 Hz, 2H), 7.51–7.48 (d, J=8.4 Hz, 2H), 7.05–7.04 (d,
J=2.1 Hz, 1H), 6.97–6.94 (d, J=9 Hz, 1H), 6.65–6.61 (dd, J=2.4, 2.4 Hz,
1H), 4.44–4.43 (d, J=2.1 Hz, 1H), 4.33–4.31 (d, J=6.3 Hz, 1H), 3.78 (s,
3H), 3.70 (s, 2H), 2.26 (s, 3H), 1.14–1.12 ppm (d, J=6.3 Hz, 3H);
13C NMR (300 MHz, CD3OD): d=172.5, 172.1, 168.4, 156.1, 138.6, 135.5,
134.1, 130.84, 130.77, 130.5, 128.7, 114.5, 113.2, 111.6, 100.7, 66.8, 57.8,
54.7, 47.0, 30.6, 19.0, 12.2 ppm (Figure S9 in the Supporting Information);
IND.HYP.GLY
Yield: 70%; m.p. 173–1748C; 1H NMR (300 MHz, CD3OD, 258C): d=
7.68–7.67 (d, J=3.6 Hz, 1H), 7.67–7.66 (d, J=3.9 Hz, 1H), 7.52–7.51 (d,
J=5.1 Hz, 2H), 7.00–6.96 (m, 1H), 6.93–6.90 (dd, J=1.8, 2.1 Hz, 1H),
6.64–6.60 (m, 1H), 4.58–4.55 (t, J=4.8 Hz, 1H), 3.90–3.61 (m, 10H), 2.22
(s, 3H), 2.19–2.11 ppm (m, 2H); 13C NMR (300 MHz, CD3OD): d=
173.0, 171.2, 170.5, 168.5, 156.1, 138.6, 135.4, 134.3, 130.9, 130.8, 128.7,
114.4, 113.0, 111.2, 101.2, 69.5, 59.2, 55.3, 37.6, 29.9, 29.2, 12.2 ppm (Fig-
ure S7 in the Supporting Information); FTIR (KBr pellet): n˜ =3375 (s,
À
FTIR (KBr pellet): n˜ =3382 (s, O H stretch), 3344 (s), 3334 (s), 1731 (m,
acid C=O stretch), 1681 (s, amide C=O stretch), 1662 (s), 1652 (s), 1525
(w), 1477 (s), 1359 (m), 1319 (s), 1222 (m), 1087 (m), 833 cmÀ1 (m);
HRMS (ESI (CH3OH)): m/z calcd for C23H23ClN2O6: 481.1142 [M+Na]+
; found: 481.1142; elemental analysis calcd (%) for C23H23ClN2O6·H2O: C
57.92, H 5.28, N 5.87; found: C 57.87, H 5.01, N 6.02%.
À
O H stretch), 1726 (w, acid C=O stretch), 1677 (s, amide C=O stretch),
1629 (s), 1608 (s), 1479 (m), 1438 (m), 1400 (w), 1357 (m), 1319 (m), 1222
(m), 1087 (m), 1033 (w), 1012 (w), 923 (w), 833 (w), 754 cmÀ1 (m);
HRMS (ESI (CH3OH)): m/z calcd for C26H26ClN3O7: 550.1357 [M+Na]+;
found: 550.1357; elemental analysis calcd (%) for C26H26ClN3O7·CH2Cl2:
C 52.91, H 4.60, N 6.86; found: C 52.32, H 4.62, N 6.59.
IND.HYP.SER
Yield: 65%; m.p. 167–1688C; 1H NMR (300 MHz, CD3OD, 258C): d=
7.70–7.66 (dd, J=4.5, 6.6 Hz, 2H), 7.54–7.51 (d, J=8.7 Hz, 2H), 7.00–
6.96 (dd, J=2.4, 2.4 Hz, 1H), 6.95–6.91 (dd, J=3.6, 3.3 Hz, 1H), 6.65–
6.62 (dd, J=2.4, 2.4 Hz, 1H), 4.63–4.58 (t, J=7.8 Hz, 1H), 4.52 (s, 1H),
4.44–4.40 (d, J=14 Hz, 1H), 3.88–3.67 (m, 9H), 2.23 (s, 3H), 2.20–
2.14 ppm (d, J=15 Hz, 2H); 13C NMR (300 MHz, CD3OD): d=173.1,
171.9, 170.7, 168.8, 156.4, 138.9, 135.7, 134.6, 131.2, 131.1, 129.0, 114.6,
113.4, 111.6, 101.5, 77.4, 69.9, 62.0, 59.5, 55.7, 55.0, 37.8, 30.1, 12.5 ppm
(Figure S4 in the Supporting Information); FTIR (KBr pellet): n˜ =3349
MTT Assay
The cytotoxicity of the IND-based hydrogelators was evaluated in RAW
264.7 cells by using a standard MTT assay.[48] The mouse macrophage
RAW 264.7 cells were purchased from American Type Culture Collection
(ATCC) and maintained by following their guidelines. The cells were
grown in Dulbeccoꢂs modified eagleꢂs medium (DMEM) supplemented
with 10% fetal bovine serum (FBS), 1% penicillin, and streptomycin in
a humidified incubator at 378C and 5% CO2. Approximately 1ꢃ104 cells/
well were seeded in 96-well plates. After 24 h of seeding, the cells were
treated with various concentrations (up to 2 mm) of hydrogelators or
DMEM alone for 24, 48, and 72 h in a humidified incubator at 378C and
5% CO2. Then, the culture medium was replaced with 100 mg of MTT
per well and kept for 4 h at 378C. To dissolve formazan produced by mi-
tochondrial reductase from live cells, DMSO (100 mL/well) was added
and incubated for 30 min at 378C. The absorbance of formazan was re-
corded at l=570 nm by using a multiplate ELISA reader (Varioskan
Flash Elisa Reader, Thermo Fisher). The percentage of live cells in the
hydrogelator-treated sample was calculated by considering the DMEM-
treated sample as 100%.
À
(s, O H stretch), 1726 (m, acid C=O stretch), 1677 (s, amide C=O
stretch), 1630 (s), 1513 (m), 1478 (m), 1322 (m), 1224 (m), 1088 (m),
755 cmÀ1 (m); HRMS (ESI (CH3OH)): m/z calcd for C27H28ClN3O8:
580.1463 [M+Na]+; found: 580.1462; elemental analysis calcd (%) for
C27H28ClN3O8·CH2Cl2: C 52.31, H 4.70, N 6.54; found: C 52.15, H 4.49, N
6.47.
IND.HYP.ALA
Yield: 71%; m.p. 154–1558C; 1H NMR (300 MHz, [D6]DMSO, 258C):
d=8.06–8.04 (d, J=6.6 Hz, 1H; N-H proton), 7.69–7.66 (d, J=9 Hz,
2H), 7.64–7.62 (d, J=6 Hz, 2H), 7.05–7.04 (d, J=2.4 Hz, 1H), 6.96–6.93
(d, J=6 Hz, 1H), 6.71–6.67 (dd, J=1.8, 1.8 Hz, 1H), 4.43–4.34 (m, 1H),
4.10–4.05 (q, J=6 Hz, 1H), 3.76 (s, 3H), 3.73–3.37 (m, 5H), 2.17 (s, 1H),
2.13 (s, 1H), 2.08 (s, 3H), 2.07–1.90 (m, 1H; O-H proton), 1.23–1.22 ppm
(d, J=2.4 Hz, 3H); 13C NMR (300 MHz, [D6]DMSO): d=175.2, 171.9,
PGE2 Assay
Production of PGE2 was estimated by using a previously published proto-
col.[49] After 24 h of seeding, about 1ꢃ106 cells were treated with
9
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Chem. Asian J. 2014, 00, 0 – 0
ꢀ 2014 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
These are not the final page numbers! ÞÞ