Medicinal Chemistry Research

, Volume 26, Issue 11, pp 2707–2717 | Cite as

Discovery two potent and new inhibitors of 15-lipoxygenase: (E)-3-((3,4-dihydroxybenzylidene) amino)-7-hydroxy-2H-chromen-2-one and (E)-O-(4-(((7-hydroxy-2-oxo-2H-chromen-3-yl) imino)methine) phenyl)dimethylcarbamothioate

  • Carolina Nuñez
  • Nicole Morales
  • Olimpo García-Beltran
  • Carolina MascayanoEmail author
  • Angelica Fierro
Original Research


The mechanisms of action and structural determinants of lipoxygenases inhibitors have been explored on several occasions, but many questions remain unanswered, especially about the differences of the inhibition mechanisms and their effect on the selectivity of lipoxygenases isoenzymes. Thus, REDOX mechanism has been proposed in this research to clarify the lipoxygenases inhibition by coumarins derivates on 15-sLOX. A series of fifteen coumarin derivatives were synthetized and evaluated as 15-lipoxygenase inhibitors. The results showed that some molecules had submicromolar activities and compete with the substrate as we observed by kinetic studies. The most relevant and interesting result was found for compound 6 who showed an inhibitory activity comparable to nordihydroguaiaretic acid a potent and REDOX inhibitor of lipoxygenases (0.17 and 0.29 μM, respectively). Finally, the docking and molecular dynamics studies showed that the better ligands were accommodated into the binding site being related with those obtained biological data. In addition, our findings contribute at the understanding of inhibitor structural requirements and elucidate the inhibition mechanism of cumarin derivatives on 15-sLOX. Thus, we point to new parameters for the future design of new ligands with potential therapeutic utility where are involved the lipoxygenases enzymes.


15-Lipoxygenase Coumarin derivatives Kinetic assay Pseudoperoxidase SAR study 



Financial support from DICYT 021641MC; Fondecyt 1120379, Fondecyt 1161375 and the Millennium Scientific Initiative (Grant P05-001-F).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

44_2017_1968_MOESM1_ESM.pdf (1.5 mb)
Supplementary Information


  1. Aliaga ME, Tiznado W, Cassels BK, Nunez MT, Millán D, Pérez EG, García-Beltrán O, Pavez P (2014) Substituent effects on reactivity of 3-cinnamoylcoumarins with thiols of biological interest. RSC Adv 4:697–704CrossRefGoogle Scholar
  2. Armstrong MM, Diaz G, Kenyon V, Holman TR (2014) Inhibitory and mechanistic investigations of oxo-lipids with human lipoxygenase isozymes. Bioorg Med Chem 22:4293–4297CrossRefPubMedPubMedCentralGoogle Scholar
  3. Bansal Y, Sethi P, Bansal G (2013) Coumarin: a potential nucleus for anti-inflammatory molecules. Med Chem Res 22:3049–3060CrossRefGoogle Scholar
  4. Bell RL, Young PR, Albert D, Lanni C, Summers JB, Brooks DW, Rubin P, Carter GW (1992) The discovery and development of zileuton: an orally active 5-lipoxygenase inhibitor. Int J Immunopharmacol 14:505–510CrossRefPubMedGoogle Scholar
  5. Berman J, Westbrook Z, Feng G, Gilliland TN, Bhat H, Weissig IN, Shindyalov PE (2000) The protein data bank. Nucleic Acids Res 28:235–242CrossRefPubMedPubMedCentralGoogle Scholar
  6. Borbulevych OY, Jankun J, Selman SH, Skrzypczak‐Jankun E (2004) Lipoxygenase interactions with natural flavonoid, quercetin, reveal a complex with protocatechuic acid in its X‐ray structure at 2.1 Å resolution. Proteins 54:13–19CrossRefPubMedGoogle Scholar
  7. Brash AR, Schneider C, Hamberg M (2012) Applications of stereospecifically-labeled fatty acids in oxygenase and desaturase biochemistry. Lipids 47:101–116CrossRefPubMedGoogle Scholar
  8. Carroll J, Jonsson EN, Ebel R, Hartman MS, Holman TR, Crews P (2001) Probing sponge-derived terpenoids for human 15-lipoxygenase inhibitors. JOC 66:6847–6851CrossRefGoogle Scholar
  9. Chauhan BF, Ducharme FM (2012) Anti‐leukotriene agents compared to inhaled corticosteroids in the management of recurrent and/or chronic asthma in adults and children. The Cochrane LibraryGoogle Scholar
  10. Cheng Y, Prusoff WH (1973) Relationship between the inhibition constant (Ki) and the concentration of inhibitor which causes 50% inhibition (I50) of an enzymatic reaction. Biochem Pharmacol 22:3099–3108CrossRefPubMedGoogle Scholar
  11. Choi J, Chon JK, Kim S, Shin W (2008) Conformational flexibility in mammalian 15S-lipoxygenase: reinterpretation of the crystallographic data. Proteins 70(3):1023–1032CrossRefPubMedGoogle Scholar
  12. Deschamps JD, Gautschi JT, Whitman S, Johnson TA, Gassner NC, Crews P, Holman TR (2007) Discovery of platelet-type 12-human lipoxygenase selective inhibitors by high-throughput screening of structurally diverse libraries. Bioorg Med Chem 15:6900–6908CrossRefPubMedPubMedCentralGoogle Scholar
  13. Frisch MJ, Trucks GW, Schlegel HB, Scuseria GE, Robb MA, Cheeseman JR, Scalmani G, Barone V, Mennucci B, Petersson GA, Nakatsuji H, Caricato M, Li X, Hratchian HP, Izmaylov AF, Bloino J, Zheng G, Sonnenberg JL, Hada M, Ehara M, Toyota K, Fukuda R, Hasegawa J, Ishida M, Nakajima T, Honda Y, Kitao O, Nakai H, Vreven T, Montgomery JA, Jr., Peralta JE, Ogliaro F, Bearpark M, Heyd JJ, Brothers E, Kudin KN, Staroverov VN, Kobayashi R, Normand J, Raghavachari K, Rendell A, Burant JC, Iyengar SS, Tomasi J, Cossi M, Rega N, Millam JM, Klene M, Knox JE, Cross JB, Bakken V, Adamo C, Jaramillo J, Gomperts R, Stratmann RE, Yazyev O, Austin AJ, Cammi R, Pomelli C, Ochterski JW, Martin RL, Morokuma K, Zakrzewski VG, Voth GA, Salvador P, Dannenberg JJ, Dapprich S, Daniels AD, Farkas Ö, Foresman JB, Ortiz JV, Cioslowski J, Fox DJ (2009) Gaussian ‘09, Revision A.1. ’09. Wallingford, CTGoogle Scholar
  14. García-Beltrán O, Cassels BK, Pérez C, Mena N, Núñez MT, Martínez NP, Aliaga ME (2014) Coumarin-based fluorescent probes for dual recognition of copper (II) and iron (III) ions and their application in bio-imaging. Sensors 14:1358–1371CrossRefPubMedGoogle Scholar
  15. García-Beltrán O, Mena N, Berríos TA, Castro EA, Cassels BK, Núñez MT, Aliaga ME (2012a) A selective fluorescent probe for the detection of mercury (II) in aqueous media and its applications in living cells. Tetrahedron Lett 53:6598–6601CrossRefGoogle Scholar
  16. García-Beltran O, Mena N, Friedrich LC, Netto-Ferreira JC, Vargas V, Quina FH, Cassels BK (2012b) Design and synthesis of a new coumarin-based ‘turn-on fluorescent probe selective for Cu+2. Tetrahedron Lett 2012(53):5280–5283CrossRefGoogle Scholar
  17. García-Beltrán O, Mena N, Yañez O, Caballero J, Vargas V, Nuñez MT, Cassels BK (2013) Design, synthesis and cellular dynamics studies in membranes of a new coumarin-based “turn-off” fluorescent probe selective for Fe 2+. Eur J Med Chem 67:60–63CrossRefPubMedGoogle Scholar
  18. García-Beltrán O, Rodríguez A, Trujillo A, Cañete A, Aguirre P, Gallego-Quintero S, Aliaga ME (2015a) Synthesis and characterization of a novel fluorescent and colorimetric probe for the detection of mercury (II) even in the presence of relevant biothiols. Tetrahedron Lett 56:5761–5766CrossRefGoogle Scholar
  19. García-Beltrán O, Rodríguez A, Trujillo A, Cañete A, Aguirre P, Gallego-Quintero S, Aliaga ME (2015b) Synthesis and characterization of a novel fluorescent and colorimetric probe for the detection of mercury (II) even in the presence of relevant biothiols. Tetrahedron Lett 56:5761–5766CrossRefGoogle Scholar
  20. Dennington R, Keith T, Millam J (2009) GaussView, Version 5 Semichem Inc., Shawnee Mission, KS,Google Scholar
  21. Giannopoulos PF, Joshi YB, Chu J, Praticò D (2013) The 12‐15‐lipoxygenase is a modulator of Alzheimer’s‐related tau pathology in vivo. Aging Cell 12:1082–1090CrossRefPubMedGoogle Scholar
  22. Gilbert NC, Bartlett SG, Waight MT, Neau DB, Boeglin WE, Brash AR, Newcomer ME (2011) The structure of human 5-lipoxygenase. Science 331:217–219CrossRefPubMedPubMedCentralGoogle Scholar
  23. Gillmor SA, Villaseñor A, Fletterick R, Sigal E, Browner MF (1997) The structure of mammalian 15-lipoxygenase reveals similarity to the lipases and the determinants of substrate specificity. Nat Struct Mol Biol 4:1003–1009CrossRefGoogle Scholar
  24. GraphPad Software, La Jolla California USA,
  25. Hadjipavlou-Litina DJ, Litinas KE, Kontogiorgis C (2007) The anti-inflammatory effect of coumarin and its derivatives. Anti-Inflamm Anti-Allergy Agents Med Chem 6:293–306CrossRefGoogle Scholar
  26. Holman TR, Zhou J, Solomon EI (1998) Spectroscopic and functional characterization of a ligand coordination mutant of soybean lipoxygenase-1: first coordination sphere analogue of human 15-lipoxygenase. JACS 120:12564–12572CrossRefGoogle Scholar
  27. Hoobler EK, Holz C, Holman TR (2013) Pseudoperoxidase investigations of hydroperoxides and inhibitors with human lipoxygenases. Bioorg Med Chem 21:3894–3899CrossRefPubMedPubMedCentralGoogle Scholar
  28. Hoult JRS, Paya M (1996) Pharmacological and biochemical actions of simple coumarins: natural products with therapeutic potential. Gener Pharmacol 27:713–722CrossRefGoogle Scholar
  29. Huff JB, Merchant BT, Mullen CR, Venkata SRT (1999) U.S. Patent No. 5,998,593. Washington, DC: U.S. Patent and Trademark OfficeGoogle Scholar
  30. Ivanov I, Heydeck D, Hofheinz K, Roffeis J, O’Donnell VB, Kuhn H, Walther M (2010) Molecular enzymology of lipoxygenases. Arch Biochem Biophys 503:161–174CrossRefPubMedGoogle Scholar
  31. Klil-Drori AJ, Ariel A (2013) 15-Lipoxygenases in cancer: a double-edged sword? Prostaglandins Other Lipid Mediat 106:16–22CrossRefPubMedGoogle Scholar
  32. Kobe MJ, Neau DB, Mitchell CE, Bartlett SG, Newcomer ME (2014) The structure of human 15-lipoxygenase-2 with a substrate mimic. J Biol Chem 289(12):8562–8569CrossRefPubMedPubMedCentralGoogle Scholar
  33. Kostova I, Saso L (2013) Advances in research of Schiff-base metal complexes as potent antioxidants. Curr Med Chem 20:4609–4632CrossRefPubMedGoogle Scholar
  34. Kotali A, Nasiopoulou DA, Tsoleridis CA, Harris PA, Kontogiorgis CA, Hadjipavlou-Litina DJ (2016) Antioxidant Activity of 3-[N-(Acylhydrazono) ethyl]-4-hydroxy-coumarins. Molecules 21:138CrossRefPubMedGoogle Scholar
  35. Krönke G, Katzenbeisser J, Uderhardt S, Zaiss MM, Scholtysek C, Schabbauer G, Baenckler HW (2009) 12/15-lipoxygenase counteracts inflammation and tissue damage in arthritis. J Immunol 183:3383–3389CrossRefPubMedGoogle Scholar
  36. Kulkarni S, Das S, Funk CD, Murray D, Cho W (2002) Molecular basis of the specific subcellular localization of the C2-like domain of 5-lipoxygenase. J Biol Chem 277:13167–13174CrossRefPubMedGoogle Scholar
  37. Li HY, Gao S, Xi Z (2009) A colorimetric and “turn-on” fluorescent chemosensor for Zn (II) based on coumarin Shiff-base derivative. Inorg Chem Commun 12:300–303CrossRefGoogle Scholar
  38. Mascayano C, Espinosa V, Sepúlveda‐Boza S, Hoobler EK, Perry S (2013) In Vitro study of isoflavones and isoflavans as potent inhibitors of human 12‐and 15‐lipoxygenases. Chem Biol Drug Des 82:317–325CrossRefPubMedGoogle Scholar
  39. Mascayano C, Espinosa V, Sepúlveda‐Boza S, Hoobler EK, Perry S, Diaz G, Holman TR (2015) Enzymatic Studies of Isoflavonoids as Selective and Potent Inhibitors of Human Leukocyte 5‐Lipo‐Oxygenase. Chem Biol Drug Des 86:114–121CrossRefPubMedGoogle Scholar
  40. Miliauskas G, Venskutonis PR, Van Beek TA (2004) Screening of radical scavenging activity of some medicinal and aromatic plant extracts. Food Chem 85:231–237CrossRefGoogle Scholar
  41. Morris GM, Goodsell DS, Huey R, Olson AJ (1996) Distributed automated docking of flexible ligands to proteins: parallel applications of AutoDock 2.4. Comput Aided Mol Des 10:293–304CrossRefGoogle Scholar
  42. Needleman P, Jakschik BA, Morrison AR, Lefkowith JB (1986) Arachidonic acid metabolism. Annu Rev Biochem 55:69–102CrossRefPubMedGoogle Scholar
  43. Newcomer ME, Brash AR (2015) The structural basis for specificity in lipoxygenase catalysis. Protein Sci 24:298–309CrossRefPubMedPubMedCentralGoogle Scholar
  44. Oldham ML, Brash AR, Newcomer ME (2005) Insights from the X-ray Crystal Structure of Coral 8R-Lipoxygenase calcium activation via a c2-like domain and a structural basis of product chirality. J Biol Chem 280:39545–39552CrossRefPubMedGoogle Scholar
  45. Phillips JC, Braun R, Wang W, Gumbart J, Tajkhorshid E, Villa E, Schulten K (2005) Scalable molecular dynamics with NAMD. J Comput Chem 26:1781–1802CrossRefPubMedPubMedCentralGoogle Scholar
  46. Ribeiro D, Freitas M, Tomé SM, Silva AM, Porto G, Cabrita EJ, Marques MM, Fernandes E (2014) Inhibition of LOX by flavonoids: a structure–activity relationship study. Eur J Med Chem 72:137–145CrossRefPubMedGoogle Scholar
  47. Riendeau D, Falgueyret JP, Guay J, Ueda N, Yamamoto S (1991) Pseudoperoxidase activity of 5-lipoxygenase stimulated by potent benzofuranol and N-hydroxyurea inhibitors of the lipoxygenase reaction. Biochem J 274:287–292CrossRefPubMedPubMedCentralGoogle Scholar
  48. Sarveswaran S, Chakraborty D, Chitale D, Sears R, Ghosh J (2015) Inhibition of 5-lipoxygenase selectively triggers disruption of c-Myc signaling in prostate cancer cells. J Biol Chem 290:4994–5006CrossRefPubMedGoogle Scholar
  49. Da Silva CM, Da Silva DL, Modolo LV, Alves RV, De Resende MA, CVB Martins, De Fátima A (2011) Schiff bases: a short review of their antimicrobial activities. J Adv Res 2:1–8CrossRefGoogle Scholar
  50. Singh D, Pathak DP (2016) Coumarins: an overview of medicinal chemistry. Potential for new drug molecules. Int J Pharm Sci Rev Res 7:482Google Scholar
  51. Skrzypczak-Jankun E, Zhou K, McCabe NP, Selman SH, Jankun J (2003) Structure of curcumin in complex with lipoxygenase and its significance in cancer. Int J Mol Med 12:17–24PubMedGoogle Scholar
  52. Somvanshi RK, Singh AK, Saxena M, Mishra B, Dey S (2008) Development of novel peptide inhibitor of Lipoxygenase based on biochemical and BIAcore evidences. BBA Proteins Proteom 1784:1812–1817CrossRefGoogle Scholar
  53. Srivastava P, Vyas VK, Variya B, Patel P, Qureshi G, Ghate M (2016) Synthesis, anti-inflammatory, analgesic, 5-lipoxygenase (5-LOX) inhibition activities, and molecular docking study of 7-substituted coumarin derivatives. Bioorg Chem 67:130–138CrossRefPubMedGoogle Scholar
  54. Tomchick DR, Phan P, Cymborowski M, Minor W, Holman TR (2001) Structural and functional characterization of second-coordination sphere mutants of soybean lipoxygenase-1. Biochemistry 40:7509–7517CrossRefPubMedGoogle Scholar
  55. Torres R, Mascayano C, Nunez C, Modak B, Faini F (2013) Coumarins Of Haplopappus multifolius and derivative as inhibitors of LOX: evaluation in-vitro and docking studies. J Chil Chem Soc 58:2027–2030CrossRefGoogle Scholar
  56. Tresaugues L, Moche M, Arrowsmith CH, Berglund H, Busam RD, Collins R, Graslund, S (2008) Crystal structure of the lipoxygenase domain of human Arachidonate 12-lipoxygenase, 12S-type. Submitted. PDB entry 3D3LGoogle Scholar
  57. Wecksler AT, Garcia NK, Holman TR (2009a) Substrate specificity effects of lipoxygenase products and inhibitors on soybean lipoxygenase-1. Bioorg Med Chem 17:6534–6539CrossRefPubMedPubMedCentralGoogle Scholar
  58. Wecksler AT, Kenyon V, Garcia NK, Deschamps JD, van der Donk WA, Holman TR (2009b) Kinetic and structural investigations of the allosteric site in human epithelial 15-lipoxygenase-2. Biochemistry 48:8721–8730CrossRefPubMedPubMedCentralGoogle Scholar
  59. Wuest SJ, Horn T, Marti-Jaun J, Kühn H, Hersberger M (2014) Association of polymorphisms in the ALOX15B gene with coronary artery disease. Clin Biochem 47:349–355CrossRefPubMedGoogle Scholar
  60. Yoshiyuki K, Hiromichi O, Shigeru A, Kimiye B, Mitsugi K (1985) Inhibition of the formation of 5-hydroxy-6, 8, 11, 14-eicosatetraenoic acid from arachidonic acid in polymorphonuclear leukocytes by various coumarins. BBA-Lipid Lipid Met 834:224–229CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  • Carolina Nuñez
    • 1
  • Nicole Morales
    • 2
  • Olimpo García-Beltran
    • 3
    • 4
  • Carolina Mascayano
    • 1
    Email author
  • Angelica Fierro
    • 2
  1. 1.Facultad de Química y Biología, Departamento de Ciencias del Ambiente, Laboratorio de Simulación Molecular y Diseño Racional de FármacosUniversidad de Santiago de ChileSantiagoChile
  2. 2.Department of Organic Chemistry, Faculty of ChemistryPontificia Universidad Católica de ChileSantiagoChile
  3. 3.Facultad de Ciencias Naturales y MatemáticasUniversidad de IbaguéIbaguéColombia
  4. 4.Department of Chemistry, Faculty of SciencesUniversity of ChileSantiagoChile

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