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Chinese Chemical Letters 19 (2008) 1419–1422

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Synthesis and anti-inflammatory-analgesic activity of

20 ,40-difluoro-3-(carbamoyl)biphenyl-4-yl benzoates
Guang Xiang Zhong *, Lu Lu Chen, Jian Li,
Wei Xiao Hu, Min You Qi
College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310032, PR China
Received 11 June 2008

Abstract
Eighteen 20 ,40 -difluoro-3-(carbamoyl)biphenyl-4-yl benzoates were synthesized from diflunisal in three steps with total yields
from 72% to 86%. All compounds were identified by IR, 1H NMR, MS and elemental analysis. The anti-inflammatory activity and
analgesic activity for 18 compounds were evaluated. The preliminary assay results showed that compounds 4a and 4p exhibited
potent anti-inflammatory-analgesic activity.
# 2008 Guang Xiang Zhong. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

Keywords: 20 ,40 -Difluoro-3-(carbamoyl)biphenyl-4-yl benzoate; Anti-inflammatory activity; Analgesic activity; Diflunisal

Fluorine-containing non-steroid anti-inflammatory drugs have been attractive for their special properties [1].
Diflunisal, 20 ,40 -difluoro-4-hydroxybiphenyl-3-carboxylic acid 1, has been approved worldwide as therapeutics for the
treatment of inflammation and pain [2]. Recently, the modifications of the chemical structure of diflunisal were
studied. For example, 3-(1,3-dihydro-2H-isoindol-2-ylcarbonyl)-20 ,40 -difluorobiphenyl-4-ol was synthesized and
reported to have the H3P-90 inhibition, and H3P-90 in abnormal cells, such as in cancer cells would damage the
regulation of signal transduction network [3]. Yu reported that esterification or amidation of diflunisal could increase
their solubility and absorption in vivo, and some of them have even better analgesic activity than that of dilunisal [4].
Some changes in the carboxyl group of diflunisal also showed good antimycobacterial, antiviral and antimicrobial
activities [5]. M.S. Roberts found that O-acyl esters of diflunisal, especially lipophilic esters, possess large
permeability surface area and tissue distribution value [6].
Our group have studied the synthesis and the SAR of fluorine-containing non-steroid anti-inflammatory drugs, and
patented that some N-substituted fluorophenylsalilylamides possessed good anti-inflammatory activity [7].
In continuation of our research, the modification of diflunisal was studied by esterification of the hydroxyl group
and amidation of the carboxylic group in diflunisal. We have synthesized 18 title compounds 4 for the evaluation of
their anti-inflammatory and analgesic activity. The synthetic route was shown as Scheme 1 starting from the

* Corresponding author.
E-mail address: drzhonggx@sohu.com (G.X. Zhong).

1001-8417/$ – see front matter # 2008 Guang Xiang Zhong. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.
doi:10.1016/j.cclet.2008.09.018
1420 G.X. Zhong et al. / Chinese Chemical Letters 19 (2008) 1419–1422

Scheme 1. Reagents and conditions: (a) pyridine, CHCl3, r.t.; (b) SOCl2, DMF, CH2Cl2, reflux; (c) NHR1R2, CH2Cl2, r.t.

Table 1
The anti-inflammatory and analgesic activities of compounds (performed by Zhejiang University of Technology).
Compound The inhibitory effect on xylene-induced mice ear The inhibitory effect on acetic acid induced mice
edema writhing
Dose (mg kg1) Inhibition (%) Dose (mg kg1) Inhibition (%)
4a 40 68.86 40 47.50
4b 40 35.90 40 15.83
4c 40 0 40 9.16
4d 40 34.07 40 37.50
4e 40 65.20 40 0
4f 40 0 40 33.33
4g 40 0 40 52.50
4p 40 43.22 40 92.50
4q 40 27.47 40 88.33
4r 40 0 40 57.50
Diflunisal 40 41.22 40 6.50

esterification of phenolic hydroxyl group and then the amidation of the carboxyl group. All the compounds 4a–r were
confirmed by IR, 1H NMR, MS and elemental analysis [8].
The biological evaluation of all compounds were carried out [9] and the results were summarized in Tables 1 and 2.
From Table 1, it could be found that in anti-inflammatory activity, there are three compounds, which possess anti-
inflammatory activity better than that of diflunisal at the dose of 40 mg kg1, especially 4a and 4e. And it could be
found that in analgesic activity, there are five compounds, which possess good analgesic activity at the dose of
40 mg kg1, especially 4p and 4q. On the whole, 4a and 4p show potent anti-inflammatory-analgesic activity.
Additionally, it indicates that the substituted groups on amide could affect the activity. It seems that the compounds
with N-alkyl group are better than that with aryl group in anti-inflammatory activity and the compounds with hetero-
atom on amide group are better than others in analgesic activity. It seems same trend that the substituted groups on the
amide could also affect on the inflammatory activity and analgesic activity. But it needs more data to find the
relationship.
 G.X. Zhong et al. / Chinese Chemical Letters 19 (2008) 1419–1422 1421

Table 2
The anti-inflammatory and analgesic activities of compounds (performed by National Center for Drug Screening).
Compound The inhibitory effect on xylene-induced mice ear The inhibitory effect on acetic acid induced mice
edema writhing
Dose (mg kg1) Inhibition (%) Dose (mg kg1) Inhibition (%)
4h 40 19.79 40 5.57
4i 40 21.40 40 20.79
4j 40 12.3 40 3.26
4k 40 0 40 0
4l 40 23.74 40 13.78
4m 40 11.14 40 0.74
4n 40 0 40 0
4o 40 5.36 40 1.43
2 40 16.06 40 7.97
Diflunisal 40 39.85 40 8.33

Acknowledgments

The authors thank the Opening Foundation of The Biochemical Engineering Key Discipline (No. 20050105),
Zhejiang, China, for financial support and the National Center for Drug Screening, Shanghai, China, for evaluation of
anti-inflammatory and analgesic activity.

References

[1] (a) H. Fujisama, T. Fujimara, Y. Takeuchi, Chem. Pharm. Bull. 53 (2005) 524;
(b) D. Bilecen, A.C. Schulte, A. Kaspar, NMR Biomed. 16 (2003) 144;
(c) G.V. Betageri, A. Nayernama, M.J. Habib, Int. J. Pharm. Adv. 1 (1996) 310 (CAN 124:193178).
[2] (a) N.S. Green, S.K. Palaninathan, J.C. Sacchettini, J. Am. Chem. Soc. 125 (2003) 13404;
(b) S.L. Adamski-werner, S.K. Palaninathan, J.C. Sacchettini, J. Med. Chem. 47 (2004) 355.
[3] L.A. Funk, M.C. Mary, P.P. Kung, WO 2006117669 (2006).
[4] C.X. Yu, L. Xu, WO 2008012603 (2008).
[5] G. Kucukguzel, A. Kocatepe, E.D. Clercq, Eur. J. Med. Chem. 41 (2006) 353.
[6] D.Y. Hung, G.D. Mellick, Y.G. Anissimov, J. Pharm. Sci. 87 (1998) 943.
[7] G.X. Zhong, K. Zhao, CN 101029010A (2007).
[8] General procedure for the synthesis of compounds 4a–r: A solution of diflunisal 1 (1 equiv.) and pyridine (2 equiv.) in CHCl3 was stirred for
30 min to protect the carboxyl group by forming a salt (mp 126–129 8C after being separated). To this reaction mixture was added dropwise a
solution of bezoyl chloride (1 equiv.) in chloroform at room temperature, and the reaction was allowed to processed with stirring for an
additional about 4 h (monitored by TLC). After that 1 mol/L HCl was used to precipitate, filtrate and dry to afford compound 2 with 95.1% yield.
It was observed that insufficient amount of pyridine would decrease the yield. A mixture of compound 2 and SOCl2 (2 equiv.) in CH2Cl2 was
refluxed for 4 h prior to work up to give compound 3. The compounds 4a–r were prepared by reaction of 3 (1 equiv.) with substituted amines
(2 equiv.) in CH2Cl2 at room temperature for 3 h, in 72–86% total yields based on the diflunisal. The data from IR, 1H NMR, MS and elemental
analysis for the compounds was listed below: The data of some compounds: 2: 95.1%, white crystalline solid, mp 167–172 8C; IR (KBr, cm1):
3444, 1738, 1689, 1267, 1210, 706; 1H NMR (500 MHz, CDCl3, d ppm): 6.96 (t, 1H, J = 8.0 Hz, 30 -H), 7.00 (t, 1H, J = 8.0 Hz, 50 -H), 7.35 (d,
1H, J = 7.5 Hz, 5-H), 7.46 (m, 1H, 60 -H), 7.50 (t, 2H, J = 8.0 Hz, 300 ,500 -H), 7.65 (t, 1H, J = 7.6 Hz, 400 -H), 7.80 (d, 1H, J = 8.0 Hz, 6-H), 8.21 (d,
2H, J = 7.5 Hz, 200 ,600 -H), 8.25 (s, 1H, 2-H), 11.24 (s, 1H, –COOH); EIMS: m/z (%) = 354 (M+, 1.02). 4a: 75.1%, white solid, mp 147–150 8C
(butanone); IR (KBr, cm(1): 3392, 1719, 1665, 1611, 1481, 1272, 1213, 709; 1H NMR (400 MHz, CDCl3, d ppm): 2.89 (d, 3H, J = 4.8 Hz, CH3),
6.36 (s, 1H, NH), 6.94 (t, 1H, J = 8.4 Hz, 30 -H), 7.00 (t, 1H, J = 8.2 Hz, 50 -H), 7.31 (d, 1H, J = 8.4 Hz, 5-H), 7.45 (m, 1H, 60 -H), 7.56 (t, 2H,
J = 7.6 Hz, 300 ,500 -H), 7.65 (d, 1H, J = 8.4 Hz, 6-H), 7.69 (t, 1H, J = 7.6 Hz, 400 -H), 7.94 (s, 1H, 2-H), 8.27 (d, 2H, J = 6.8 Hz, 200 ,600 -H); EIMS: m/
z (%) = 367 (M+, 1.36); Anal. Calcd. for C21H15F2NO3: C, 68.66; H, 4.12; N, 3.81. Found: C, 68.49; H, 4.14; N, 3.70. 4d: 72.3%, white
crystalline, mp 123–128 8C (butanone); IR (KBr, cm(1): 1733, 1647, 1478, 1263, 1215, 708; 1H NMR (400 MHz, CDCl3, d ppm): 2.95, 3.03 (s, s,
6H, J = 4.80 Hz, CH3), 6.93 (t, 1H, J = 8.4 Hz, 30 -H), 6.97 (t, 1H, J = 8.2 Hz, 50 -H), 7.42 (d, 1H, J = 8.0 Hz, 5-H), 7.42 (m, 1H, 60 -H), 7.51 (s, 1H,
2-H), 7.51 (t, 2H, J = 7.6 Hz, 300 ,500 -H), 7.59 (d, 1H, J = 8.0 Hz, 6-H), 7.65 (t, 1H, J = 7.6 Hz, 400 -H), 8.17 (d, 2H, J = 7.6 Hz, 200 ,600 -H); EIMS: m/z
(%) = 381 (M+, 15.57); Anal. Calcd. for C22H17F2NO3: C, 69.29; H, 4.49; N, 3.67. Found: C, 69.27; H, 4.46; N, 3.65. 4e: 72.0%, white solid, mp
125–128 8C (butanone); IR (KBr, cm(1): 1739, 1633, 1471, 1266, 1211, 710; 1H NMR (400 MHz, CDCl3, d ppm): 1.02, 1.08 (t, t, 6H, J = 7.2 Hz,
CH3), 3.24 (m, 4H, CH2), 6.94 (t, 1H, J = 8.4 Hz, 30 -H), 6.98 (t, 1H, J = 8.4 Hz, 50 -H), 7.43 (d, 1H, J = 8.4 Hz, 5-H), 7.43 (m, 1H, 60 -H), 7.50
(t, 3H, J = 7.6 Hz, 2,300 ,500 -H), 7.58 (d, 1H, J = 8.0 Hz, 6-H), 7.64 (t, 1H, J = 8.0 Hz, 400 -H), 8.18 (d, 2H, J = 7.2 Hz, 200 ,600 -H); EIMS: m/z
(%) = 409 (M+, 4.78); Anal. Calcd. for C24H21F2NO3: C, 70.41; H, 5.17; N, 3.42. Found: C, 70.48; H, 5.08; N, 3.30. 4g: 81.3%, white powder,
1422 G.X. Zhong et al. / Chinese Chemical Letters 19 (2008) 1419–1422

mp 150–154 8C (butanone); IR (KBr, cm(1): 3268, 1741, 1643, 1594, 1485, 1267, 1211, 701; 1H NMR (400 MHz, CDCl3, d ppm): 4.52 (d, 2H,
J = 5.2 Hz, CH2), 6.66 (s, 1H, NH), 6.94 (t, 1H, J = 8.0 Hz, 30 -H), 6.98 (t, 1H,J = 8.0 Hz, 50 -H), 7.14 (m, 5H, Ar000 H), 7.29 (d, 1H, J = 8.4 Hz, 5-
H), 7.46 (m, 1H, 60 -H), 7.47 (t, 2H, J = 7.6 Hz, 300 ,500 -H), 7.66 (t, 1H, J = 7.2 Hz, 400 -H), 7.67 (d, 1H, J = 7.6 Hz, 6-H), 8.03 (s, 1H, 2-H), 8.07 (d,
2H, J = 7.2 Hz, 200 ,600 -H); EIMS: m/z (%) = 443 (M+, 1.00); Anal. Calcd. for C27H19F2NO3: C, 73.13; H, 4.32; N, 3.16. Found: C, 73.18; H, 4.49;
N, 3.08. 4h: 84.1%, white solid, mp 139–140 8C (butanone); IR (KBr, cm(1): 3335, 1745, 1647, 1597, 1442, 1265, 1200, 707; 1H NMR
(400 MHz, CDCl3, d ppm): 6.96 (t, 1H,J = 8.4 Hz, 30 -H), 7.00 (t, 1H, J = 8.4 Hz, 50 -H), 7.08 (t, 1H, J = 7.6 Hz, 4000 -H), 7.26 (t, 2H, J = 8.0 Hz,
3000 ,5000 -H), 7.37 (d, 1H, J = 8.0 Hz, 5-H), 7.45 (d, 2H, J = 7.6 Hz, 2000 ,6000 -H), 7.47 (m, 1H, 60 -H), 7.55 (t, 2H, J = 7.6 Hz, 300 ,500 -H), 7.69 (t, 1H,
J = 7.6 Hz, 400 -H), 7.72 (d, 1H, J = 8.0 Hz, 6-H), 8.10 (s, 1H, 2-H), 8.23 (s, 1H, NH), 8.24 (d, 2H, J = 7.2 Hz, 200 ,600 -H); EIMS: m/z (%) = 429
(M+, 0.89); Anal. Calcd. for C26H17F2NO3: C, 72.72; H, 3.99; N, 3.26. Found: C, 72.59; H, 4.01; N, 3.19. 4i: 85.9%, white powder, mp 165–
168 8C (butanone); IR (KBr, cm(1): 3445, 1736, 1644, 1602, 1487, 1272, 1213, 706; 1H NMR (400 MHz, CDCl3, d ppm): 2.18 (s, 3H, CH3), 6.96
(t, J = 8.4 Hz, 1H, 30 -H), 7.00 (t, 1H, J = 8.0 Hz, 50 -H), 7.07 (t, 1H, J = 7.2 Hz, 4000 -H), 7.14 (d, 1H, J = 7.6 Hz, 3000 -H), 7.20 (t, 1H, J = 6.8 Hz,
5000 -H), 7.36 (d, 1H, J = 8.4 Hz, 5-H), 7.48 (m, 1H, 60 -H), 7.53 (t, 2H, J = 7.6 Hz, 300 ,500 -H), 7.68 (t, 1H, J = 7.6 Hz, 400 -H), 7.72 (d, 1H, J = 8.4 Hz,
6-H), 7.85 (d, 1H, J = 8.4 Hz, 6000 -H), 7.90 (s, 1H, -NH), 8.09 (s, 1H, 2-H), 8.22 (d, 2H, J = 7.2 Hz, 200 ,600 -H); EIMS: m/z (%) = 443 (M+, 0.33);
Anal. Calcd. for C27H19F2NO3: C, 73.13; H, 4.32; N, 3.16. Found: C, 73.27; H, 4.40; N, 3.13. 4j: 85.9%, white powder, mp 130–133 8C
(butanone); IR (KBr, cm(1): 3317, 1745, 1646, 1611, 1486, 1265, 1204, 708; 1H NMR (400 MHz, CDCl3, d ppm): 2.28 (s, 3H, CH3), 6.90 (d, 1H,
J = 7.6 Hz, 4000 -H), 6.96 (t, J = 8.4 Hz, 30 -H), 7.00 (t, 1H, J = 8.0 Hz, 50 -H), 7.14 (t, 1H, J = 8.0 Hz, 5000 -H), 7.24 (d, 1H, J = 8.0 Hz, 6000 -H), 7.25
(s, 1H, 2000 -H), 7.38 (d, 1H, J = 8.4 Hz, 5-H), 7.48 (m, 1H, 60 -H), 7.55 (t, 2H, J = 8.4 Hz, 300 ,500 -H), 7.68 (t, 1H, J = 7.6 Hz, 400 -H), 7.71 (d, 1H,
J = 8.4 Hz, 6-H), 8.11 (s, 1H, 2-H), 8.20 (s, 1H, NH), 8.25 (d, 2H, J = 7.2 Hz, 200 , 600 -H); EIMS: m/z (%) = 443 (M+, 1.25); Anal. Calcd. for
C27H19F2NO3: C, 73.13; H, 4.32; N, 3.16. Found: C, 73.18; H, 4.36; N, 3.10. 4l: 85.3%, white powder, mp 174–176 8C (butanone); IR (KBr,
cm(1): 3430, 1744, 1646, 1600, 1485, 1268, 1215, 702; 1H NMR (400 MHz, CDCl3, d ppm): 2.10 (s, 3H, 2000 -CH3), 2.27 (s, 3H, 5000 -CH3), 6.87 (d,
1H, J = 7.6 Hz, 4000 -H), 6.96 (t, 1H, J = 8.4 Hz, 30 -H), 6.99 (t, 1H, J = 8.0 Hz, 50 -H), 7.00 (d, 1H, J = 7.2 Hz, 3000 -H), 7.35 (d, 1H, J = 8.4 Hz, 5-H),
7.48 (m, 1H, 60 -H), 7.51 (t, 2H, J = 8.0 Hz, 300 ,500 -H), 7.61 (s, 1H, 6000 -H), 7.66 (t, 1H, J = 7.6 Hz, 400 -H), 7.71 (d, 1H, J = 7.6 Hz, 6-H), 7.83 (s, 1H,
NH), 8.07 (s, 1H, 2-H), 8.21 (d, 2H, J = 7.2 Hz, 200 , 600 -H); EIMS: m/z (%) = 457 (M+, 0.88); Anal. Calcd. for C28H21F2NO3: C, 73.51; H, 4.63; N,
3.06. Found: C, 73.67; H, 4.75; N, 2.97. 4m: 82.0%, white needle crystal, mp 139–141 8C (butanone); IR (KBr, cm(1): 3407, 1735, 1649, 1593,
1481, 1270, 1214, 704; 1H NMR (400 MHz, CDCl3, d ppm): 6.96 (t, 1H, J = 8.4 Hz, 30 -H), 6.99 (t, 1H, J = 8.0 Hz, 50 -H), 7.03 (t, 1H, J = 7.2 Hz,
4000 -H), 7.26 (d, 1H, J = 7.2 Hz, 3000 -H), 7.29 (t, 1H, J = 8.0 Hz, 5000 -H), 7.37 (d, 1H, J = 8.4 Hz, 5-H), 7.49 (m, 1H, 60 -H), 7.51 (t, 2H, J = 7.6 Hz,
300 ,500 -H), 7.66 (t, 1H, J = 7.2 Hz, 400 -H), 7.74 (d, 1H, J = 8.4 Hz, 6-H), 8.14 (s, 1H, 2-H), 8.23 (d, 2H, J = 7.6 Hz, 200 ,600 -H), 8.47 (d, 1H,
J = 7.6 Hz, 6000 -H), 8.67 (s, 1H, NH); EIMS: m/z (%) = 463 (M+, 0.37); Anal. Calcd. for C26H16ClF2NO3: C, 67.32; H, 3.48; N, 3.02. Found: C,
67.28; H, 3.53; N, 2.93. 4p: 83.5%, yellow solid, mp 184–185 8C (butanone); IR (KBr, cm(1): 3343, 1705, 1665, 1601, 1484, 1271, 1206, 709; 1H
NMR (400 MHz, CDCl3, d ppm): 3.79 (s, 3H, –CH3), 6.79 (d, 2H, J = 8.4 Hz, 3000 ,5000 -H), 6.95 (t, 1H, J = 8.4 Hz, 30 -H), 6.99 (t, 1H, J = 7.6 Hz,
50 -H), 7.34 (d, 1H, J = 8.0 Hz, 5-H), 7.35 (d, 3H, J = 8.0 Hz, 2000 ,6000 -H), 7.47 (m, 1H, 60 -H), 7.54 (t, 2H, J = 7.6 Hz, 300 ,500 -H), 7.69 (t, 1H,
J = 8.0 Hz, 400 -H), 7.70 (d, 1H, J = 8.4 Hz, 6-H), 8.09 (s, 1H, 2-H), 8.12 (s, 1H, NH), 8.24 (d, 2H, J = 8.0 Hz, 200 ,600 -H); EIMS: m/z (%) = 459
(M+, 6.79); Anal. Calcd. for C27H19F2NO4: C, 70.58; H, 4.17; N, 3.05. Found: C, 70.38; H, 4.19; N, 3.18. 4q: 82.9%, white crystalline solid, mp
154–157 8C (butanone); IR (KBr, cm(1): 1735, 1634, 1462, 1260, 1208, 708; 1H NMR (500 MHz, CDCl3, d ppm): 2.20 (s, 3H, –CH3), 2.28 (t,
4H, J = 4.2 Hz, 3000 -CH2), 3.38, 3.72 (t, t, 4H, J = 4.8 Hz, 2000 -CH2), 6.94 (t, 1H, J = 8.0 Hz, 30 -H), 6.97 (t, 1H, J = 8.0 Hz, 50 -H), 7.42 (d, 1H,
J = 8.4 Hz, 5-H), 7.42 (m, 1H, 60 -H), 7.49 (s, 1H, 2-H), 7.51 (t, 2H, J = 8.0 Hz, 300 ,500 -H), 7.60 (d, 1H, J = 8.0 Hz, 6-H), 7.65 (t, 1H, J = 7.6 Hz,
400 -H), 8.18 (d, 2H, J = 8.0, 200 -H); EIMS: m/z (%) = 436 (M+, 6.91); Anal. Calcd. for C25H23F2N3O3: C, 66.51; H, 5.13; N, 9.31. Found: C,
66.37; H, 5.30; N, 9.16. 4r: 83.1%, white powder, mp 169–172 8C (butanone); IR (KBr, cm(1): 1724, 1633, 1487, 1271, 1211, 706; 1H NMR
(400 MHz, CDCl3, d ppm): 3.45 (m, 4H, 2000 ,6000 -CH2), 3.60 (m, 4H, 3000 ,5000 -CH2), 6.95 (t, 1H, J = 8.0 Hz, 30 -H), 7.00 (t, 1H, J = 8.0 Hz, 50 -H),
7.42 (d, 1H, J = 8.4 Hz, 5-H), 7.43 (m, 1H, 60 -H), 7.53 (s, 1H, 2-H), 7.55 (t, 2H, J = 7.6, 300 ,500 -H), 7.63 (d, 1H, J = 8.0 Hz, 6-H), 7.69 (t, 1H,
J = 7.6 Hz, 400 -H), 8.21 (d, 2H, J = 7.6 Hz, 200 ,600 -H); EIMS: m/z (%) = 423 (M+, 6.70); Anal. Calcd. for C24H19F2NO4: C, 68.08; H, 4.52; N,
3.31. Found: C, 67.99; H, 4.54; N, 3.29.
[9] Evaluation of the biological activities. The anti-inflammatory activity was evaluated in a xylene-induced mice ear edema model. The Kunming
mice (weight 18–22 g for Table 1 and weight 20–26 g for Table 2) chosen were orally administrated with selected compounds at the dose of
40 mg kg1. And the analgesic activity was tested in an acetic acid-induced mice writhing model. Diflunisal was introduced as a positive control.