https://doi.org/10.22037/ijpr.2013.1326

2-(4-Fluorophenyl)-N-phenylacetamide Derivatives as Anticancer Agents: Synthesis and In-vitro Cytotoxicity Evaluation

authors:

avatar Alireza Aliabadi a , * , avatar Sajad Andisheh a , b , avatar Zahra Tayarani-Najaran c , avatar Mona Tayarani-Najaran d

Department of Medicinal Chemistry, Faculty of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Students Research Committee, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
Departmrnt of Chemistry, Alzahra University, Tehran, Iran.
Iranian Journal of Pharmaceutical Research : IJPR: Vol.12, issue 3; 267-271
article type: Original Article
received: February , 2012
accepted: June , 2012

How To Cite Aliabadi A, Andisheh S, Tayarani-Najaran Z, Tayarani-Najaran M. 2-(4-Fluorophenyl)-N-phenylacetamide Derivatives as Anticancer Agents: Synthesis and In-vitro Cytotoxicity Evaluation. Iran J Pharm Res. 2013;12(3):e125688. https://doi.org/10.22037/ijpr.2013.1326.

Abstract

Cancer is a major global problem and is the second leading cause of mortality in the developed countries.Resistance to current chemotherapeutics and high incidence of adverse effects are the two principal reasons for developing new anticancer agents. Phenylacetamide derivatives can act as potential anticancer agents. Synthesis and screening of 2-(4-Fluorophenyl)- N-phenylacetamide derivatives in present study showed that these compounds act as potent anticancer agents especially against PC3(prostate carcinoma) cell line. Compounds 2a-2c with nitro moiety demonstrated a higher cytotoxic effect than compounds 2d-2f with methoxy moiety. All compounds in this series exhibited lower activity than imatinib as reference drug. Compounds 2b (IC50 = 52 μM) and 2c (IC50 = 80 μM) were the most active compounds against PC3 cell line in comparison with imatinib(IC50 = 40 μM). Compound 2c (IC50 = 100 μM) with p-nitro substituent was the most active compound compared to imatinib(IC50 = 98 μM) in MCF-7 cell line.

Introduction

Cancer is a major global problem and is the second leading cause of mortality in the developed countries. Since many of the current pharmacotherapeutic methods have problems with toxicity and drug-resistance, there is a strong demand for the discovery and development of effective, saferand novelcancer therapies (1).

Since it is known that the anti-tumor efficacy of many chemotherapeutic agents are correlated with their ability to induce apoptosis, novel approaches to promote apoptosis in cancer cells via targeting regulators of apoptosis could lead to the development of new anticancer treatments. In addition, these new agents may overcome tumor resistance to the conventional anti-cancer drugs (2-9).

Phenylacetate (PA) and related aromatic fatty acids have been shown to possess anti-proliferative and differentiating effects on various human cancer cell lines such as glioblastomas, leukemias, prostate carcinomas, and breast carcinomas. A phase I clinical trial with PA has also proved in-vivo anti-tumor activity in humans. Moreover, it has been reported that PA induces the differentiation process together with apoptosis in-vitro and in-vivo. The ability of PA to induce tumor growth inhibition, differentiation, and apoptosis of cancer cells, along with the relatively low toxicity at clinically effective doses, prompted us to find a more potent analogue of PA. Recently, several derivatives of phenyl acetic acid were synthesized and reported as potential anticancer agents (Figure 1) (10, 11).

Structure of 4-Fluoro-N-butylphenylacetamide as potent anticancer lead compound

Structure of 4-Fluoro-N-butylphenylacetamide as potent anticancer lead compound

Figure. 1

On the other hand, aniline derivatives are also potent in-vitro inhibitors of cell proliferation and growth (Figure 2) (12-14). In the present project, we combined the structure of phenylacetamide and anilide derivatives for designing of new anticancer agents (Figure 3).

Structure of two aniline derivatives as potent anticancer lead compounds

Structure of two aniline derivatives as potent anticancer lead compounds

Figure 2

Design of 2-(4-Fluorophenyl)-N-phenylacetamide derivatives

Design of 2-(4-Fluorophenyl)-N-phenylacetamide derivatives

Figure 3

Compounds 2a-2f were tested against three cancerous cell lines using MTS assay. Cell lines used in this study werePC3(prostate carcinoma), MCF-7(breast cancer) and HL- 60(promyelocytic leukemia) (Table 1). Totally, all tested compounds showed a better cytotoxic activity toward the PC3 cell line in comparison with other cell lines. On the other hand, MCF-7 cell line was the most resistant cell line to the tested compounds. Compounds 2a- 2c with nitro moiety demonstrated a higher cytotoxic effect than compounds 2d-2f with methoxy moiety. All compounds in this series exhibited lower activity than imatinib as reference drug. Compounds 2b (IC50 = 52 μM) and 2c (IC50 = 80 μM) were the most active compounds against PC3 cell line in comparison with imatinib (IC50 = 40 μM). Compound 2c (IC50 = 100 μM) with p-nitro substituent was the most active compound compared to imatinib(IC50 = 98 μM) in MCF-7 cell line.

Table 1

Cytotoxicity results(IC50, μM) of compounds 2a-2g in comparison with imatinib

2a
2b
2c
2d
2e
2f
2g
Imatinib
o-NO2m-NO2p-NO2o-OCH3m-OCH3p-OCH3H
PC31965280158156168250<40
MCF-7250<191250<250<247250<250<79
HL-60208178100218206243250<98

Experimental

General procedure for the synthesis of compounds 2a-2g

According to the Figure 3, for the preparation of compounds 2a-2g, equimolar quantities of 4-fluorophenyl acetic acid, EDC and HOBt were mixed and stirred in acetonitrile for 30 min. Then, the appropriate aniline derivatives were added and stirring was continued for 24 h. The completion of the reaction was checked by thin layer chromatography. The acetonitrile was evaporated and water/ethyl acetate was added. Ethyl acetate phase was separated and washed two times by sodium bicarbonate, diluted sulfuric acid and brine. The organic layer was dried by anhydrous sodium sulfate and filtered. The ethyl acetate was evaporated under reduced pressure using rotary evaporator (14).

2-(4-Fluorophenyl)-N-(2-nitrophenyl) acetamide (2a)

mp: 122-129 °C, Yield: 65%, 1H NMR (CDCl3, 400 MHz) δ: 3.63 (s, 2H, -CH2-), 6.69(t, 1H, J = 8Hz, H4-2-Nitrophenyl), 6.8 (d, 1H, J = 8Hz, H6-2-Nitrophenyl), 7.2 (t, 1H, J = 8Hz, H3-2-Nitrophenyl), 7.54 (t, 2H, J = 8Hz, H2,6, 4-Fluorophenyl), 7.89(t, 2H, J = 8Hz, H3,5, 4-Fluorophenyl), 8.11(d, 1H, J = 8Hz, H3-2- Nitrophenyl), 10.15 (brs, 1H, NH). IR(KBr, cm- 1) ῡ: 3475, 3344, 1624, 1494, 1431, 1342, 1220, 1157, 720. MS(m/z, %): 274(M+, 25), 136(75), 109(100), 83(20), 63(10).

2-(4-Fluorophenyl)-N-(3-nitrophenyl) acetamide(2b)

mp: 138 °C, Yield: 57%,1H NMR (CDCl3, 400 MHz) δ: 3.76 (s, 2H, -CH2-), 6.65 (d, 2H, J = 8Hz, 4-Fluorophenyl), 6.85 (d, 2H, J = 8Hz, 4-Fluorophenyl), 7.07 (t, 1H, J = 8Hz, H5-3-Nitrorophenyl), 7.15 (d, 1H, H6-3- Nitrophenyl), 7.26 (d, 1H, H4-3-Nitrophenyl), 7.29(s, 1H, H2-3-Nitrophenyl), 10.15 (brs, 1H, NH). IR(KBr, cm-1) δ: 2922, 1624, 1523, 1508, 1344, 1259, 731. MS(m/z, %): 274(M+, 20), 136(85), 109(100), 83(22), 63(8).

2-(4-Fluorophenyl)-N-(4-nitrophenyl) acetamide(2c)

mp: 123 °C, Yield: 72%,1H NMR (CDCl3, 400 MHz) δ:3.76(s, 2H, -CH2CO-), 6.62(d, 2H, J = 8Hz, 4-Flurophenyl), 7.62(d, 2H, J = 8Hz, 4-Fluorophenyl), 8.07(d, 2H, J = 8Hz, H2,6 , 4-Nitrophenyl), 8.17(d, 2H, J = 8 Hz, H3,5 , 4-Nitrophenyl), 10.16 (brs, 1H, NH). IR(KBr, cm-1) δ: 3340, 1712, 1597, 1543, 1500, 1338, 1253, 1147, 1111, 858, 750. MS(m/z, %): 274(M+, 20), 136(80), 109(100), 83(25), 63(10).

2-(4-Fluorophenyl)-N-(2-methoxyphenyl) acetamide(2d)

mp: 98 °C, Yield: 61%,1H NMR (CDCl3, 400 MHz) δ: 3.72 (s, 2H, -CH2-), 3.76 (s, 3H, -OCH3), 6.81 (d, 1H, J = 8 Hz, H3-2-Methoxyphenyl), 6.94 (t, 1H, J = 8Hz, H4-2-Methoxyphenyl), 7.01 (t, 1H, J = 8Hz, H5-2-Methoxyphenyl), 7.06(t, 2H, J = 8Hz, H2,6-4-Fluorophenyl), 7.30(t, 2H, J = 8Hz, H3,5-4-Fluorophenyl), 7.76 (brs, 1H, NH), 8.33 (d, 1H, J = 8 Hz, H6-2-Methoxyphenyl). IR (KBr, cm-1) δ: 3282, 2900, 2860, 1662, 1596, 1537, 1510, 1500, 1460, 1436, 1409, 1342, 1259, 1232, 1215, 1182, 1157, 1114, 1031, 950, 752. MS(m/z, %): 259(M+, 35), 123(65), 109(100), 83(30).

2-(4-Fluorophenyl)-N-(3-methoxyphenyl) acetamide(2e)

mp:108-110 °C, Yield: 56 %,1H NMR (CDCl3, 400 MHz) δ: 3.69 (s, 2H, -CH2-), 3.77 (s, 3H, -OCH3), 6.64 (d, 1H, J = 8 Hz, H4-3-Methoxyphenyl), 6.84 (d, 1H, J = 8 Hz, H6-3-Methoxyphenyl), 7.08(m, 3H, aromatic), 7.16(t, 2H, J = 8Hz, 4-Fluorophenyl), 7.25(t, 2H, J = 8Hz, 4-Fluorophenyl), 7.6 (brs, 1H, NH).IR(KBr, cm-1) δ: 3260, 1664, 1597, 1533, 1510, 1452, 1429, 1288, 1219, 1155, 1035, 779. MS(m/z, %): 259(M+, 20), 150(90), 123(85), 109(100), 92(40), 83(30), 77(40), 52(30).

2-(4-Fluorophenyl)-N-(4-methoxyphenyl)acetamide(2f)

mp:142-148 °C, Yield: 78%,1H NMR (DMSO-d6, 400 MHz) δ: 3.68 (s, 2H, -CH2), 3.77 (s, 3H, -OCH3), 6.82 (d, 2H, J = 8 Hz, H3,5, 4-Methoxyphenyl), 7.06 (t, 2H, H3,5, 4-Fluorophenyl), 7.30 (t, 2H, H2,6, 4-Fluorophenyl), 7.36 (d, 2H, J = 8 Hz, H2,6, 4-Methoxyphenyl), 7.55 (brs, 1H, NH). IR(KBr, cm-1) δ: 3300, 1651, 1604, 1539, 1506, 1492, 1452, 1436, 1409, 1246, 1234, 1219, 1157, 1031, 827, 754. MS(m/z, %): 259(M+, 40), 123(100), 108(95), 83(20).

2-(4-Fluorophenyl)-N-phenylacetamide(2g)

mp:131-134 °C, Yield: 69%,1H NMR (DMSO-d6, 400 MHz) δ: 3.63(s, 2H, -CH2-), 7.03(t, 1H, J = 8 Hz, H4-phenyl), 7.15(t, 2H, J = 8 Hz, H2,6-4-Fluorophenyl), 7.29(t, 2H, J = 8 Hz, H3,5, 4-Fluorophenyl), 7.36(t, 2H, J = 8 Hz, H3,5-Phenyl), 7.58(d, 2H, J = 8 Hz, H2,6-Phenyl). IR(KBr, cm-1) δ: 3284, 1654, 1598, 1546, 1508, 1487, 1442, 1220, 1130, 752. MS(m/z, %): 229(M+, 20), 136(10), 120(40), 109(95), 93(100), 83(40), 77(20), 65(30).

Cytotoxicity assay

The MTS assay is based on the reduction by mitochondrial dehydrogenase in metabolically active cells of the novel tetrazolium compound, MTS, to the water-soluble formazan that absorbs at 490 nm. PC3, MCF7 and HL-60 cells were seeded in each well of a 96-microwell plate and treated with various concentrations of the compound(s) to be tested. After incubation for 48 h, Cell Titer 96 Aqueous One Solution Reagent (Promega, Madison, WI), which is composed of the novel tetrazolium compound MTS and an electron coupling reagent phenazineethosulfate (PES, a redox intermediary), was added to each well according to the manufacturer’s instructions. After 3 h, cell viability was determined by measuring the absorbance at the wavelength of 490 nm using an ELISA microplate reader (Awareness, Palm City, FL). The cytotoxicity of the compound(s) was presented as mean of 3 independent experiments with 3 replicates for each compound (s) concentration (15).

Acknowledgements

References

  • 1.

    Kemnitzer W, Kuemmerle J, Jiang S, Zhang HZ, Sirisoma N, Kasibhatla SC, Crogan-Grundy B, Tseng J, Drewe S, Cai X. Discovery of 1-benzoyl-3-cyanopyrrolo[1,2-a]quinolines as a new series of apoptosis inducers using a cell- and caspase-based high-throughput screening assay. Part 1: Structure–activity relationships of the 1- and 3-positions. Bioorg. Med. Chem. Lett. 2008;18:6259-6264. [PubMed ID: 18952423].

  • 2.

    Kemnitzer W, Kuemmerle J, Zhang HZ, Kasibhatla S, Tseng B, Drewe J, Cai SX. Discovery of 3-aryl-5-aryl-1,2,4-oxadiazoles as a new series of apoptosis inducers. 2. Identification of more aqueous soluble analogs as potential anticancer agents. Bioorg. Med. Chem. Lett. 2009;15:4410-4415. [PubMed ID: 19500976].

  • 3.

    Earnshaw WC, Martins LM, Kaufmann SH. Mammalian caspases: structure, activation, substrates, and functions during apoptosis. Annu. Rev. Biochem. 1999;68:383-424. [PubMed ID: 10872455].

  • 4.

    Bratton SB, MacFarlane M, Cain K, Cohen GM. Protein complexes activate distinct caspase cascades in death receptor and stress-activated apoptosis. Exp. Cell Res. 2000;256:27-33. [PubMed ID: 10739648].

  • 5.

    Hirata H, Takahashi A, Kobayashi S, Yonehara S, Sawai H, Okazaki T, Yamamoto K, Sasada M. Caspases are activated in a branched protease cascade and control distinct downstream processes in Fas-induced apoptosis. J. Exp. Med. 1998;187:587-600. [PubMed ID: 9463409].

  • 6.

    Jiang S, Crogan-Grundy C, Drewe J, Tseng B, Cai SX. Discovery of (naphthalen-4-yl) (phenyl)methanones and N-methyl-N-phenylnaphthalen-1-amines as new apoptosis inducers using a cell- and caspase-based HTS assay. Bioorg. Med. Chem. Lett. 2008;18:5725-5728. [PubMed ID: 18849165].

  • 7.

    Bortner CD, Cidlowski JA. A necessary role for cell shrinkage in apoptosis. Biochem. Pharmacol. 1998;56:1549-1559. [PubMed ID: 9973175].

  • 8.

    Mirian M, Zarghi A, Sadeghi S, Tabaraki P, Tavallaee M, Dadrass O, Sadeghi-aliabadi H. Synthesis and cytotoxic evaluation of some novel sulfonamide derivatives against a few human cancer cells. Iran. J. Pharm. Res. 2011;10:741-748.

  • 9.

    Mokhtari S, Mosaddegh M, Hamzeloo Moghadam M, Soleymani Z, Ghafari S, Kobarfard F. Synthesis and cytotoxic evaluation of novel 3-substituted derivatives of 2-indolinone. Iran. J. Pharm. Res. 2011;11:411-421.

  • 10.

    Chan HC, Kuo SC, Liu SC, Liu CH, Hsu SL. 4-Fluoro-N-butylphenylacetamide: a synthetic phenylacetate derivative that upregulates Bcl-XS, activates caspase cascade and induces apoptosis in human squamous lung cancer CH27 cells. Cancer Lett. 2002;186:211-221. [PubMed ID: 12213291].

  • 11.

    Liu CH, Huang LJ, Kuo SC, Lee TH, Kan-Jen Tsai c, Hsu-Chin Chan. 4-Fluoro-N-butylphenylacetamide (H6) inhibits cell growth via cell-cycle arrest and apoptosis in human cervical cancer cells. Bioorg. Med. Chem. Lett. 2009;17:42-48.

  • 12.

    Wan-Ping H, Hsin-Su Y, Yan-Ren C, Yi-Min T, Yin-Kai C, Chao-Cheng L, Long-Sen C, Jeh-Jeng W. Synthesis and biological evaluation of thiobenzanilides as anticancer agents. Bioorg. Med. Chem. 2008;9:5295-5302.

  • 13.

    Xiong Cai S, Nguyen B, Jia S, Herich J, Guastella J, Reddy S, Tseng B, Drewe J, Kasibhatla S. Discovery of substituted N-phenyl nicotinamide as potent inducer of apoptosis using a cell- and caspase-based high throughput screening assay. J. Med. Chem. 2003;46:2474-2481. [PubMed ID: 12773051].

  • 14.

    Aliabadi A, Shamsa F, Ostad SN, Emami S, Shafiee A, Davoodi J, Foroumadi A. Synthesis and biological evaluation of 2-phenylthiazole-4-carboxamide derivatives as anticancer agents. Eur. J. Med. Chem. 2010;11:5384-5389. [PubMed ID: 20846760].

  • 15.

    Malich G, Markovic B, Winder C. The sensitivity and specificity of the MTS tetrazolium assay for detecting the in-vitro cytotoxicity of 20 chemicals using human cell lines. Toxicol. 1997;124:179-192.