Pyrrolizine-5-carboxamides: Exploring the impact of various substituents on anti-inflammatory and anticancer activities

Open access


Towards optimization of the pyrrolizine-5-carboxamide scaffold, a novel series of six derivatives (4a-c and 5a-c) was prepared and evaluated for their anti-inflammatory, analgesic and anticancer activities. The (EZ)-7-cyano-6-((4-hydroxybenzylidene)amino)-N-(p-tolyl)-2,3-dihydro-1H-pyrrolizine-5-carboxamide (4b) and (EZ)-6-((4-chlorobenzylidene)-amino)-7-cyano-N-(p-tolyl)-2,3-dihydro-1H-pyrrolizine-5-carboxamide (5b) bearing the electron donating methyl group showed the highest anti-inflammatory activity while (EZ)-6-((4-chlorobenzylidene)amino)-7-cyano-N-phenyl-2,3-dihydro-1H-pyrrolizine-5-carboxamide (5a) was the most active analgesic agent. Cytotoxicity of the new compounds was evaluated against the MCF-7, A2780 and HT29 cancer cell lines using the MTT assay. Compounds 4b and 5b displayed high anticancer activity with IC50 in the range of 0.30–0.92 μmol L−1 against the three cell lines, while compound (EZ)-N-(4-chlorophenyl)-7-cyano-6-((4-hydroxybenzylidene)-amino)-2,3-dihydro-1H-pyrrolizine-5-carboxamide (4c) was the most active against MCF-7 cells (IC50 = 0.08 μmol L−1). Both the anti-inflammatory and anticancer activities of the new compounds were dependent on the type of substituent on the phenyl rings. Substituents with opposite electronic effects on the two phenyl rings are preferable for high cytotoxicity against the MCF-7 and A2780 cells. COX inhibition was suggested as the molecular mechanism of the anti-inflammatory activity of the new compounds while no clear relationship could be observed between COX inhibition and anticancer activity. Compound 5b, the most active against the three cell lines, induced dose-dependent early apoptosis with 0.1–0.2 % necrosis in MCF-7 cells. New compounds showed promising drug-likeness scores while the docking study revealed high binding affinity to COX-2. Taken together, this study highlighted the significant impact of the substituents on the anti-inflammatory and anticancer activity of pyrrolizine-5-carboxamides, which could help in further optimization to discover good leads for the treatment of cancer and inflammation.

1. E. R. Rayburn, S. J. Ezell and R. Zhang, Anti-inflammatory agents for cancer therapy, Mol. Cell. Pharmacol. 1 (2009) 29–43;

2. M. J. Thun, S. J. Henley and C. Patrono, Nonsteroidal anti-inflammatory drugs as anticancer agents: mechanistic, pharmacologic, and clinical issues, J. Natl. Cancer Inst. 94 (2002) 252–266;

3. S. R. Pedada, N. S. Yarla, P. J. Tambade, B. L. Dhananjaya, A. Bishayee, K. M. Arunasree, G. H. Philip, G. Dharmapuri, G. Aliev, S. Putta and G. Rangaiah, Synthesis of new secretory phospholipase A2-inhibitory indole containing isoxazole derivatives as anti-inflammatory and anticancer agents, Eur. J. Med. Chem. 112 (2016) 289–297;

4. C. Sobolewski, C. Cerella, M. Dicato, L. Ghibelli and M. Diederich, The role of cyclooxygenase-2 in cell proliferation and cell death in human malignancies, Int. J. Cell Biol. 2010 (2010) 1–21;

5. C. S. Williams, M. Mann and R. N. DuBois, The role of cyclooxygenases in inflammation, cancer, and development, Oncogene 18 (1999) 7908–7916;

6. C. Ruegg, J. Zaric and R. Stupp, Non-steroidal anti-inflammatory drugs and COX-2 inhibitors as anti-cancer therapeutics: hypes, hopes and reality, Ann. Med. 35 (2003) 476–487;

7. A. T. Koki and J. L. Masferrer, Celecoxib: a specific COX-2 inhibitor with anticancer properties, Cancer Control 9 (2002) 28–35;

8. A. M. Gouda and A. H. Abdelazeem, An integrated overview on pyrrolizines as potential anti-inflammatory, analgesic and antipyretic agents, Eur. J. Med. Chem. 114 (2016) 257–292;

9. A. M. Gouda, A. H. Abdelazeem, H. A. Omar, A. N. Abdalla, M. A. S. Abourehab and H. I. Ali, Pyrrolizines: design, synthesis, anticancer evaluation and investigation of the potential mechanism of action, Bioorg. Med. Chem. 25 (2017) 5637–5651; (in press)

10. A. M. Gouda, H. I. Ali, W. H. Almalki, M. A. Azim, M. A. S. Abourehab and A. H. Abdelazeem, Design, synthesis, and biological evaluation of some novel pyrrolizine derivatives as COX inhibitors with anti-inflammatory/analgesic activities and low ulcerogenic liability, Molecules 21 (2016) 1–21;

11. J.-P. Raynauld, J. Martel-Pelletier, P. Bias, S. Laufer, B. Haraoui, D. Choquette, A. D. Beaulieu, F. Abram, M. Dorais, E. Vignon and J.-P. Pelletier, Protective effects of licofelone, a 5-lipoxygenase and cyclo-oxygenase inhibitor, versus naproxen on cartilage loss in knee osteoarthritis: a first multicentre clinical trial using quantitative MRI, Ann. Rheum. Dis. 68 (2009) 938–947;

12. W. Liu, J. Zhou, K. Bensdorf, H. Zhang, H. Liu, Y. Wang, H. Qian, Y. Zhang, A. Wellner, G. Rubner, W. Huang, C. Guo and R. Gust, Investigations on cytotoxicity and anti-inflammatory potency of licofelone derivatives, Eur. J. Med. Chem. 46 (2011) 907–913;

13. S. Tavolari, M. Bonafe, M. Marini, C. Ferreri, G. Bartolini, E. Brighenti, S. Manara, V. Tomasi, S. Laufer and T. Guarnieri, Licofelone, a dual COX/5-LOX inhibitor, induces apoptosis in HCA-7 colon cancer cells through the mitochondrial pathway independently from its ability to affect the arachidonic acid cascade, Carcinogenesis 29 (2008) 371–380;

14. S. Tavolari, A. Munarini, G. Storci, S. Laufer, P. Chieco and T. Guarnieri, The decrease of cell membrane fluidity by the non-steroidal anti-inflammatory drug Licofelone inhibits epidermal growth factor receptor signalling and triggers apoptosis in HCA-7 colon cancer cells, Cancer Lett. 321 (2012) 187–194;

15. V. Lisowski, C. Enguehard, J. Lancelot, D. Caignard, S. Lambel, S. Leonce, A. Pierre, G. Atassi, P. Renard and S. Rault, Design, synthesis and antiproliferative activity of tripentones: a new series of antitubulin agents, Bioorg. Med. Chem. Lett. 11 (2001) 2205–2208;

16. V. Lisowski, S. Leonce, L. Kraus-Berthier, J. Sopkova-de Oliveira Santos, A. Pierre, G. Atassi, D.-H. Caignard, P. Renard and S. Rault, Design, synthesis, and evaluation of novel thienopyrrolizinones as antitubulin agents, J. Med. Chem. 47 (2004) 1448–1464;

17. C. Rochais, T. Cresteil, V. Perri, M. Jouanne, A. Lesnard, S. Rault and P. Dallemagne, MR22388, a novel anti-cancer agent with a strong FLT-3 ITD kinase affinity, Cancer Lett. 331 (2013) 92–98;

18. A. M. Gouda, A. H. Abdelazeem, E.-S. A. Arafa and K. R. A. Abdellatif, Design, synthesis and pharmacological evaluation of novel pyrrolizine derivatives as potential anticancer agents, Bioorg. Chem. 53 (2014) 1–7;

19. A. Etienne and Y. Correia, Derivatives of 2-pyrrolidone, Bull. Soc. Chem. 10 (1969) 3704–3712.

20. W. A. Jacobs and M. Heidelberger, The ferrous sulfate and ammonia method for the reduction of nitro to amino compounds, J. Am. Chem. Soc. 39 (1917) 1435–1439;

21. M. Y. Ebeid, S. M. El-Moghazy, M. M. Hanna, F. A. Romeih and F. F. Barsoum, Synthesis and anti-HIV activity of some 6,7-dihydro-5H-pyrrolizine-3-carboxamide, 5,6,7,8-tetrahydroindolizine-3-carboxamide, 1-thioxo-1,2,3,5,6,7,8,9,10,11-decahydro-pyrimido-[1,6-a]azonine-4-carbonitrile and 6-thioxo-1,2,5,6,8,9,10,11,12,13,14,14a-dodecahydro-pyrimido[4ʹ,5ʹ:4,5]pyrimido-[1,6-a]azonine-1-one derivatives, Bull. Fac. Pharm. Cairo Univ. 35 (1997) 171–183.

22. C. A. Winter, E. A. Risley and G. W. Nuss, Carrageenan induced edema in hind paw of the rats as an assay for anti-inflammatory drugs, Proc. Soc. Exp. Biol. Med. 111 (1962) 544–547;

23. A. Mollica, R. Costante, A. Stefanucci, F. Pinnen, G. Lucente, S. Fidanza and S. Pieretti, Novel cyclic biphalin analogue with improved antinociceptive properties, J. Pept. Sci. 19 (2013) 233–239;

24. N. Handler, W. Jaeger, H. Puschacher, K. Leisser and T. Erker, Synthesis of novel curcumin analogues and their evaluation as selective cyclooxygenase-1 (COX-1) inhibitors, Chem. Pharm. Bull. (Tokyo) 55 (2007) 64–71;

25. E.-S. A. Arafa, A. H. Abdelazeem, H. H. Arab and H. A. Omar, OSU-CG5, a novel energy restriction mimetic agent, targets human colorectal cancer cells in vitro, Acta Pharmacol. Sin. 35 (2014) 394–400;

26. I. Vermes, C. Haanen, H. Steffens-Nakken and C. Reutellingsperger, A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V, J. Immunol. Methods 184 (1995) 39–51;

27. C. A. Lipinski, F. Lombardo, B. W. Dominy and P. J. Feeney, Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings, Adv. Drug Deliv.Rev. 23 (1997) 3–25;

28. A. Daina, O. Michielin and V. Zoete, SwissADME: a free web tool to evaluate pharmacokinetics, drug-likeness and medicinal chemistry friendliness of small molecules, Sci. Rep. 7 (2017) Article ID 42717;

29. B. S. Selinsky, K. Gupta, C. T. Sharkey and P. J. Loll, Structural analysis of NSAID binding by prostaglandin H2 synthase: time-dependent and time-independent inhibitors elicit identical enzyme conformations, Biochemistry 40 (2001) 5172–5180;

30. R. G. Kurumbail, A. M. Stevens, J. K. Gierse, J. J. McDonald, R. A. Stegeman, J. Y. Pak, D. Gildehaus, J. M. Miyashiro, T. D. Penning, K. Seibert, P. C. Isakson and W. C. Stallings, Structural basis for selective inhibition of cyclooxygenase-2 by anti-inflammatory agents, Nature 384 (1996) 644–648;

Acta Pharmaceutica

The Journal of Croatian Pharmaceutical Society

Journal Information

IMPACT FACTOR 2017: 1.071
5-year IMPACT FACTOR: 1.623

CiteScore 2017: 1.46

SCImago Journal Rank (SJR) 2017: 0.362
Source Normalized Impact per Paper (SNIP) 2017: 0.642


All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 123 123 82
PDF Downloads 75 75 51