t, J 8.4, ArH), 6.77 (2H, d, J 9.0, ArH) and 3.13 (6H, s, 2 ×
NCH3,); dC(75 MHz, CDCl3) 170.0, 166.3, 156.4, 153.2, 145.9,
143.9, 133.1, 132.6, 131.1, 130.5, 126.5, 125.8, 122.1, 120.3, 115.0,
111.6 and 40.4.
172.8, 167.0, 155.4, 152.9, 143.7, 134.2, 128.1, 125.6, 122.4, 111.6,
57.6, 52.4, 40.4, 31.8, 19.2 and 18.2.
General procedure for the preparation of 7i–k. Procedure C.
N-[[4-(p-Dimethylaminophenylazo)]benzoyl]-benzotriazole (3a)
(200 mg, 0.54 mmol) and corresponding amine (2.0–3.0 eq.), Et3N
(0.23mL, 1.62 mmol) were mixed in DMF (10mL). The mixture
was heated at 150 ◦C for 5–10 h. After the evaporation of solvent
under reduced pressure, the residue was worked up with MeOH
and the products were obtained as red solids.
General method for the preparation of carboxylic azo-dye labeled
amino acids 5a–m. N-(4-Arylazo)benzoyl-benzotriazole
3
(200 mg, 0.54 mmol for 3a and 0.611 mmol for 3b, 1eq.) and
amino acid 4 (1eq.) were added to a mixture of DMF and water
(3 : 1,v/v), and stirred at room temperature for 24 h. After the
evaporation of solvent, washed with CH2Cl2 and drying under
vacuum, the corresponding pure products 5 were obtained with
high yield of 81–99%; For 5k–m, after the evaporation of solvent,
the residue was dissolved in CH2Cl2 and washed with 4 N HCl.
4-[4-(Dimethylamino)phenylazo]benzoyl-N-methylaniline (7i).
3 eq. 6i; worked up with MeOH to give 7i as red microcrystal
(140 mg, 74%); mp 185.0–187.0 ◦C (from MeOH); found: C,
73.38; H, 6.35; N, 15.71. Calc. for C22H22N4O: C, 73.72; H, 6.19;
N, 15.63%; dH(300 MHz; DMSO-d6; Me4Si) 7.75 (2H, d, J 7.5,
ArH), 7.57 (2H, d, J 6.9, ArH), 7.38 (2H, d, J 7.8, ArH), 7.27
(2H, d, J 6.6, ArH), 7. 20 (3H, d, J 6.3, ArH), 6.82 (2H, d, J
7.8, ArH), 3.40 (3H, s, CONCH3) and 3.06 (6H, s, 2 × NCH3);
dC(75 MHz; DMSO-d6; Me4Si) 168.9, 152.7, 152.4, 144.4, 142.6,
136.8, 129.3, 129.1, 127.1, 126.5, 125.0, 121.0, 111.5, 39.8 and
37.8.
4-[(4-Dimethylamino)phenylazo]benzoyl-glycine (5a). (175 mg,
99%). Redmi◦crocrystal;mp 238.0–240.0 ◦C (from CH2Cl2) (lit.,4b,10
mp 232–233 C); dH(300 MHz, DMSO-d6; Me4Si) 8.94 (1H, t, J
5.7, NH), 8.02 (2H, d, J 8.7, ArH), 7.84 (2H, d, J 8.7, ArH), 7.83
(2H, d, J 9.3, ArH), 6.85 (2H, d, J 9.3, ArH), 3.95 (2H, d, J 6.0,
CH2) and 3.08 (6H, s, 2 × CH3); dC(75 MHz, DMSO-d6) 171.4,
165.9, 154.1, 152.9, 142.7, 134.1, 128.4, 125.1, 121.6, 111.6, 41.4
and 39.8.
Gerenal procedure for the preparation of 7a–f. Procedure A.
N-[[4-(p-Dimethylaminophenylazo)]benzoyl]-benzotriazole (3)
(200 mg, 0.54 mmol for 3a and 0.611 mmol for 3b) and
corresponding amines (1–3eq.) were mixed in THF (10mL) and
stirred for 1–48 h at room temperature (monitored by TLC). After
the evaporation of solvent, pure products 7 were obtained from
the residues after simple purification procedures with high yield
of 85–100%.
Acknowledgements
We thank Dr C. D. Hall for helpful discussions.
Notes and references
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9; (c) C. Renner and L. Moroder, ChemBioChem, 2006, 7, 868; (d) I.
Willner, Acc. Chem. Res., 1997, 30, 347.
4-[4-(Dimethylamino)phenylazo]benzoyl-morpholine (7a). 1eq.
6a; purified with column on silica gel eluting with ethyl acetate–
hexane (1 : 2, v/v) to give 7a as red microcrystal (184 mg, 100%);
mp 212.0–215.0 ◦C (from EtOAc–hexane); found: C, 67.57; H,
6.66; N, 16.57. Calc. for C19H22N4O2: C, 67.44; H, 6.55; N, 15.56%;
dH(300 MHz, CDCl3) 7.88 (4H, t, J 8.1, ArH), 7.52 (2H, d, J 8.1,
ArH), 6.76 (2H, d, J 9.0, ArH), 3.85–3.40 (8H, br s, CH2CH2) and
3.11 (6H, s, 2 × CH3); dC(75 MHz, CDCl3) 170.1, 154.0, 152.8,
143.6, 135.6, 128.1, 125.4, 122.3, 111.5, 66.9, 40.3 and 29.8.
2 (a) Z. Shen and Y. Zhang, Synth. Commun., 2000, 30, 2525; (b) A. D.
Abell, M. A. Jones, A. T. Neffe, S. G. Aitken, T. P. Cain, R. J. Payne, S. B.
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Morton, J. Med. Chem., 2007, 50, 2916; (c) G. T. Wang, C. C. Chung,
T. F. Holzmann and G. A. Krafft, Anal. Biochem., 1993, 210, 351; (d) R.
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The preparation of 7g, 7h. Procedure B. N-[[4-(p-Dimethyl-
aminophenylazo)]benzoyl]-benzotriazole
(3a)
(200
mg,
0.54 mmol) was added to the solution of amine (1–3eq.)
and Et3N (3.0 eq.) in THF (10 ml) at room temperature. The
mixture was heated under reflux for 24 h. After filtration, the
solvent was evaporated under reduced pressure. Pure products
were obtained from the residues after simple purification
procedures.
4-[4-(Dimethylamino)phenylazo]benzoyl-L-valine methyl ester
(7g). 1.0eq. 6g; purified with column on silica gel eluting with
ethyl acetate–hexanes (1 : 3, v/v) to give 7g as red microcrystal
◦
(188 mg, 90%); mp 156.0–158.0 C (from EtOAc–hexane); [a]D25
+59.6 (c 2.8 in MeOH); retention time: 3.29; found: C, 66.05; H,
6.94; N, 14.37. Calc. for C21H26N4O3: C, 69.95; H, 6.85; N, 14.65%;
dH(300 MHz, CDCl3) 7.90 (6H, m, ArH), 6.76 (2H, d, J 9.0, ArH),
6.67 (1H, d, J 8.4, NH), 4.83–4.79 (1H, m, J 5.1, 3.6, NHCH),
3.79 (3H, s, OCH3), 3.12 (6H, s, 2 × NCH3), 2.33–2.27 (1H, m,
CHCH3) and 1.02 (6H, t, J 6.6, 2 × CHCH3); dC(75 MHz, CDCl3)
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(c) T. N. Grossmann and O. Seitz, J. Am. Chem. Soc., 2006, 128, 15596;
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The Royal Society of Chemistry 2008
Org. Biomol. Chem., 2008, 6, 2400–2404 | 2403
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