Open Access Peer-reviewed Research Article

Main Article Content

Eliane Blanco Nunes
Priscila Zei Melo
Jefté Barbosa
Jefferson Hollanda Veras
Carolina Ribeiro e Silva
Wanderlene Blanco Nunes
Lee Chen-Chen
Caridad Noda Pérez
Stanislau Parreira Cardozo
Aline Bernardes corresponding author
Elisângela de Paula Silveira Lacerda

Abstract

The chalcones (E)-3-(4-chlorophenyl)-1-phenyl-2-propen-1-one (4-CL) and (E)-3-(3,4-dimethoxyphenyl)-1-phenyl-2 -propen-1-one (DMF) are versatile and easily synthesized into low-cost compounds that have a wide spectrum of biological activities. In this study, the cytotoxic, genotoxic and modulatory activities of 4-CL and DMF were evaluated using the Ames test and the mouse micronucleus assay. The results of the Ames test revealed that both chalcones did not show mutagenic activity in Salmonella typhimurium strains TA98 and TA100, and demonstrated significant antimutagenicity (p< 0.05) when co-administered with sodium azide (SA) in strain TA100. In the micronucleus assay, both showed a significant increase in the frequency of micronucleated polychromatic erythrocytes (MNPCE) at 24 h and 48 h, revealing a genotoxic effect. In the co-treatment with mitomycin C (MMC) there was a significant decrease (p< 0.05) in the frequency of MNPCE both in chalcones at 24h and in the less concentrated dose of DMF at 48h, demonstrating its antigenotoxic activity. 4-CL showed a significant decrease in the polychromatic/ normochromatic erythrocyte (PCE/ NCE) ratio at 24 and 48 h (p< 0.05), indicating cytotoxicity. However, 4-CL and DMF when co-administered with MMC showed a significant increase in the PCE/NCE ratio within 24 hours, demonstrating anticytotoxicity. Furthermore, a biphasic dose-response behavior was observed in both chalcones, 4-CL in the co-administration with SA, in the Ames Test and DMF in the co-treatment with MMC, at 48 hours of exposure, in the micronucleus assay. In this study, 4-CL and DMF showed genotoxic, cytotoxic, antigenotoxic, anticytotoxic and no mutagenic properties.

Keywords
Salmonella typhimurium, mutagenicity, genotoxicity, chlorochalcone, dimethoxychalcone, micronucleus, OECD

Article Details

Supporting Agencies
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Apoio à Pesquisa de Goiás (FAPEG) and Coordenação de Aperfeiçoamento de Pessoal de Ensino Superior - Brazil (CAPES)
How to Cite
Nunes, E., Melo, P., Barbosa, J., Veras, J., e Silva, C., Nunes, W., Chen-Chen, L., Pérez, C., Cardozo, S., Bernardes, A., & Lacerda, E. de P. (2022). In vitro and in vivo evaluation of genotoxicity, cytotoxicity, and protective effects of synthetic chalcones (E)-3-(4-chlorophenyl)-1-phenyl-2-propen-1-one (4-CL) and (E)-3-(3,4-dimethoxyphenyl)-1- phenyl-2-propen-1-one (DMF). Journal of Pharmaceutical and Biopharmaceutical Research, 3(2), 206-217. https://doi.org/10.25082/JPBR.2021.01.005

References

  1. Zhuang C, Zhang W, Sheng C, et al. Chalcone: A privileged structure in medicinal chemistry. Chemical Reviews, 2017, 117(12): 7762–7810. https://doi.org/10.1021/acs.chemrev.7b00020
  2. Jasim HA, Nahar L, Jasim MA, et al. Chalcones: Synthetic Chemistry Follows Where Nature Leads. Biomolecules, 2021, 11(8): 1203. https://doi.org/10.3390/BIOM11081203
  3. Rammohan A, Reddy JS, Sravya G, et al. Chalcone synthesis, properties and medicinal applications: a review. Environmental Chemistry Letters, 2020, 1–26. https://doi.org/10.1007/s10311-019-00959-w
  4. Kar Mahapatra D, Asati V and Bharti SK. An updated patent review of therapeutic applications of chalcone derivatives (2014-present). Expert Opinion on Therapeutic Patents, 2019, 29(5): 385–406. https://doi.org/10.1080/13543776.2019.1613374
  5. Adelusi TI, Akinbolaji GR, Yin X, et al. Neurotrophic, anti-neuroinflammatory, and redox balance mechanisms of chalcones. European Journal of Pharmacology, 2021, 891: 173695. https://doi.org/10.1016/j.ejphar.2020.173695
  6. Moreira J, Almeida J, Saraiva L, et al. Chalcones as Promising Antitumor Agents by Targeting the p53 Pathway: An Overview and New Insights in Drug-Likeness. Molecules, 2021, 26(12): 3737.
  7. Rocha S, Ribeiro D, Fernandes E, et al. A Systematic Review on Anti-diabetic Properties of Chalcones. Current Medicinal Chemistry, 2020, 27(14): 2257–2321. https://doi.org/10.2174/0929867325666181001112226
  8. Rani A, Anand A, Kumar K, et al. Recent developments in biological aspects of chalcones: the odyssey continues. Expert Opinion on Drug Discovery, 2019, 14(3): 249–288. https://doi.org/10.1080/17460441.2019.1573812
  9. Salehi B, Quispe C, Chamkhi I, et al. Pharmacological Properties of Chalcones: Efficacy, Molecular Mechanisms, Pharmacological Properties of Chalcones: A Review of Preclinical Including Molecular Mechanisms and Clinical Evidence. Frontiers in Pharmacology, 2021, 11. https://doi.org/10.3389/fphar.2020.592654
  10. Elkhalifa D, Al-hashimi I, Moustafa A Al, et al. A comprehensive review on the antiviral activities of chalcones. Journal of Drug Targeting, 2021, 29(4): 403–419. https://doi.org/10.1080/1061186X.2020.1853759
  11. López SN, Castelli MV, Zacchino SA, et al. In vitro antifungal evaluation and structure-activity relationships of a new series of chalcone derivatives and synthetic analogues, with inhibitory properties against polymers of the fungal cell wall. Bioorganic and Medicinal Chemistry, 2001, 9(8): 1999–2013. https://doi.org/10.1016/S0968-0896(01)00116-X
  12. Talniya NC and Sood P. Synthesis and antimicrobial activity of chalcones. Journal of Chemical and Pharmaceutical Research, 2016, 8(5): 610–613. https://doi.org/ISSN:0975-7384
  13. Kunthalert D, Baothong S, Khetkam P, et al. A chalcone with potent inhibiting activity against biofilm formation by nontypeable haemophilus influenzae. Microbiology and Immunology, 2014, 58(10): 581–589. https://doi.org/10.1111/1348-0421.12194
  14. Alam MS, Rahman SMM and Lee DU. Synthesis, biological evaluation, quantitative-SAR and docking studies of novel chalcone derivatives as antibacterial and antioxidant agents. Chemical Papers, 2015, 69(8): 1118–1129. https://doi.org/10.1515/chempap-2015-0113
  15. Sangwan M and Pathak DP. Synthesis and biological activity of chalcone derivatives as anti - asthmatics agents. Chemical Science Transactions, 2016, 5(3): 579–586. https://doi.org/10.7598/cst2016.1232
  16. Ramalho SD, Bernades A, Demetrius G, et al. Synthetic chalcone derivatives as inhibitors of cathepsins K and B, and their cytotoxic evaluation. Chemistry and Biodiversity, 2013, 10(11): 1999–2006. https://doi.org/10.1002/cbdv.201200344
  17. Chen G, Xie W, Nah J, et al. 3,4-Dimethoxychalcone induces autophagy through activation of the transcription factors TFE3 and TFEB. https://doi.org/10.15252/emmm.201910469
  18. Kumar A, Rajmohan TP and Ragavan K. Evaluation of anti-inflammatory , antioxidant and antiproliferative activities of halogenated chalcones. World Journal of Pharmacy and Pharmaceutical Sciences, 2016, 5(5): 978–1001. https://doi.org/10.20959/wjpps20165-6667
  19. Lunardi F, Guzela M, Rodrigues AT, et al. Trypanocidal and leishmanicidal properties of substituition - containing chalcones. Antimicrobial Agents and Chemotherapy, 2003, 47(4): 1449–1451. https://doi.org/10.1128/AAC.47.4.1449
  20. Corrêa R, Fenner BP, Buzzi FDC, et al. Antinociceptive activity and preliminary structure-activity relationship of chalcone-like compounds. Zeitschrift fur Naturforschung - Section C Journal of Biosciences, 2008, 63(11–12): 830–836.
  21. Begum NA, Roy N, Laskar RA, et al. Mosquito larvicidal studies of some chalcone analogues and their derived products: Structure-activity relationship analysis. Medicinal Chemistry Research, 2011, 20(2): 184–191. https://doi.org/10.1007/s00044-010-9305-6
  22. Targanski SK, Sousa JR, de P´adua GMS, et al. Larvicidal activity of substituted chalcones against Aedes aegypti (Diptera: Culicidae) and non-target organisms. Pest Management Science, 2021, 77(1): 325–334. https://doi.org/10.1002/PS.6021
  23. Ortolan XR, Fenner BP, Mezadri TJ, et al. Osteogenic potential of a chalcone in a critical-size defect in rat calvaria bone. Journal of Cranio-Maxillofacial Surgery, 2014, 42(5): 520–524. https://doi.org/10.1016/j.jcms.2013.07.020
  24. Gonz´alez LA, Escobar G, Upegui YA, et al. Effect of substituents in the A and B rings of chalcones on antiparasite activity. Archiv der Pharmazie, 2020, 353(12): 2000157. https://doi.org/10.1002/ardp.202000157
  25. Pati HN, Das U, Sakagami H, et al. 1,3-diaryl-2-propenones and 2-benzylidene-1,3-indandiones: a quest for compounds displaying greater toxicity to neoplasms than normal cells. Archiv der Pharmazie (Weinheim), 2010, 343(9): 535–541. https://doi.org/10.1002/ardp.200900308.1
  26. Sahin ID, Christodoulou MS, Guzelcan EA, et al. A small library of chalcones induce liver cancer cell death through Akt phosphorylation inhibition. Scientific Reports, 2020, 10(1): 1–9. https://doi.org/10.1038/s41598-020-68775-9
  27. OECD. Test guideline 471, Genetic toxicology: bacterial reverse mutation test. Ninth addendum to the OECD Guidelines for the testing of chemicals 1997. Paris: Organization for Economic Cooperation and Development.
  28. OECD. Test guideline 474, Mammalian erythrocyte micronucleus test. OECD Guidelines for the testing of chemicals 2016. Paris: Organization for Economic Cooperation and Development.
  29. Mortelmans K. Mutat Res Gen Tox En A perspective on the development of the Ames Salmonella / mammalian- microsome mutagenicity assay. Mutation research-genetic toxicology and environmental mutagenesis, 2019, 841(March): 14–16. https://doi.org/10.1016/j.mrgentox.2019.04.004
  30. Mortelmans K and Zeiger E. The Ames Salmonella/microsome mutagenicity assay. Mutation Research/ Fundamental and Molecular Mechanisms of Mutagenesis, 2000, 455(1): 29–60. https://doi.org/10.1016/S0027-5107(00)00064-6
  31. Levy DD, Zeiger E, Escobar PA, et al. Mutat Res Gen Tox En Recommended criteria for the evaluation of bacterial mutagenicity data (Ames test). Mutation research-genetic toxicology and environmental mutagenesis, 2019, 848: 403074. https://doi.org/10.1016/j.mrgentox.2019.07.004
  32. Fenech M, Dellios J, Fenech M, et al. The in vitro micronucleus technique Related papers The in vitro micronucleus technique. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, 2000, 455(1–2): 81–95.
  33. Krishna G and Hayashi M. In vivo rodent micronucleus assay: protocol, conduct and data interpretation. Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, 2000, 455(1–2): 155–166. https://doi.org/10.1016/S0027-5107(00)00117-2
  34. Maron DM and Ames BN. Revised methods for the Salmonella mutagenicity test. Mutation Research, 1983, 113(3–4): 173–215. https://doi.org/10.1016/0165-1161(83)90010-9
  35. Guvenalp Z, Ozbek H, Karadayi M, et al. Two antigenotoxic chalcone glycosides from Mentha longifolia subsp. longifolia. Pharmaceutical Biology, 2015, 53(6): 888–896. https://doi.org/10.3109/13880209.2014.948633
  36. Gulluce M, Agar G, Baris O, et al. Mutagenic and antimutagenic effects of hexane extract of some Astragalus species grown in the Eastern Anatolia region of Turkey. Phytotherapy Research, 2010, 24(7): 1014–1018. https://doi.org/10.1002/ptr.3059
  37. Horn RC and Vargas VMF. Antimutagenic activity of extracts of natural substances in the Salmonella/ microsome assay. Mutagenesis, 2003, 18(2): 113–118. https://doi.org/10.1093/mutage/18.2.113
  38. Cabrera M, Lavaggi ML, Croce F, et al. Identification of chalcones as in vivo liver monofunctional phase II enzymes inducers. Bioorganic and Medicinal Chemistry, 2010, 18(14): 5391–5399. https://doi.org/10.1016/j.bmc.2010.05.033
  39. Kr¨amer A, Pudil J, Frank H, et al. Some substrates and inhibitors of cytosolic epoxide hydrolase induce mutations in Salmonella typhimurium and V79 cells. Mutation Research, 1993, 290(2): 165–174. https://doi.org/10.1016/0027-5107(93)90156-A
  40. Lapchak PA. Drug-like property profiling of novel neuroprotective compounds to treat acute ischemic stroke: Guidelines to develop pleiotropic molecules. Translational Stroke Research, 2013, 4(3): 328–342. https://doi.org/10.1007/s12975-012-0200-y
  41. Lima DCDS, Vale CR do, Ve´eras JH, et al. Absence of genotoxic effects of the chalcone (E)-1- (2-hydroxyphenyl)-3-(4-methylphenyl)-prop-2-en-1-one) and its potential chemoprevention against DNA damage using in vitro and in vivo assays. PLoS ONE, 2017, 12(2): 1-15. https://doi.org/10.1371/journal.pone.0171224
  42. Rashid KA, Mullin CCA and Mumma RRO. Structure-mutagenicity relationships of chalcones and their oxides in the Salmonella assay. Mutation Research, 1986, 169(3): 71-79. https://doi.org/10.1016/0165-1218(86)90086-8
  43. Nazir S, Ansari FL, Hussain T, et al. Brine shrimp lethality assay “an effective prescreen”: Microwaveassisted synthesis, BSL toxicity and 3DQSAR studies-based designing, docking and antitumor evaluation of potent chalcones. Pharmaceutical Biology, 2013, 51(9): 1091-1103. https://doi.org/10.3109/13880209.2013.777930
  44. Cabrera M, Simoens M, Falchi G, et al. Synthetic chalcones, flavanones, and flavones as antitumoral agents: Biological evaluation and structure-activity relationships. Bioorganic and Medicinal Chemistry, 2007, 15(10): 3356-3367. https://doi.org/10.1016/j.bmc.2007.03.031
  45. Karamana I, Gezegenb H, G¨urdereb MB, et al. Screening of biological activities of a series of chalcone derivatives against human pathogenic microorganisms. Chemistry & Biodiversity, 2010, 7(2): 400–408. https://doi.org/10.1002/cbdv.200900027
  46. Ritter M, Martins RM, Rosa SA, et al. Green synthesis of chalcones and microbiological evaluation. Journal of the Brazilian Chemical Society, 2015, 26(6): 1201-1210. https://doi.org/10.5935/0103-5053.20150084
  47. Ventura TLB, Calixto SD, Abrahim-Vieira B de A, et al. Antimycobacterial and anti-inflammatory activities of substituted chalcones focusing on an anti-tuberculosis dual treatment approach. Molecules, 2015, 20(5): 8072-8093. https://doi.org/10.3390/molecules20058072
  48. Ansari FL, Nazir S, Noureen H, et al. Combinatorial synthesis and antibacterial evaluation of an indexed chalcone library. Chemistry and Biodiversity, 2005, 2(12): 1656-1664. https://doi.org/10.1002/cbdv.200590135
  49. Konda RK, Nuthakki VK and Babu M. P. Synthesis, characterization and biological screening of novel substituted chalcones. Der Pharmacia Lettre, 2014, 6(4): 1-4. https://doi.org/ISSN:0975-5071
  50. Go M, Wu X and Liu X. Chalcones: An update on cytotoxic and chemoprotective properties. Current Medicinal Chemistry, 2005, 12(4): 483–499. https://doi.org/10.2174/0929867053363153
  51. Mirzaei S, Hadizadeh F, Eisvand F, et al. Synthesis, structure-activity relationship and molecular docking studies of novel quinoline-chalcone hybrids as potential anticancer agents and tubulin inhibitors. Journal of Molecular Structure, 2020, 1202: 127310. https://doi.org/10.1016/j.molstruc.2019.127310
  52. Pereira D, Lima RT, Palmeira A, et al. Design and synthesis of new inhibitors of p53–MDM2 interaction with a chalcone scaffold. Arabian Journal of Chemistry, 2019, 12(8): 4150-4161. https://doi.org/10.1016/j.arabjc.2016.04.015
  53. Xu F, Li W, Shuai W, et al. Design, synthesis and biological evaluation of pyridine-chalcone derivatives as novel microtubule-destabilizing agents. European Journal of Medicinal Chemistry, 2019, 173: 1-14. https://doi.org/10.1016/j.ejmech.2019.04.008
  54. Iftikhar S, Khan S, Bilal A, et al. Synthesis and evaluation of modified chalcone based p53 stabilizing agents. Bioorganic and Medicinal Chemistry Letters, 2017, 27(17): 4101-4106. https://doi.org/10.1016/j.bmcl.2017.07.042
  55. Sotibr´an ANC, Ordaz-T´ellez MG and Rodr´ıguez-Arnaiz, R. Flavonoids and oxidative stress in Drosophila melanogaster. Mutation Research - Genetic Toxicology and Environmental Mutagenesis, 2011, 726(1): 60-65. https://doi.org/10.1016/j.mrgentox.2011.08.005
  56. Rozmer Z and Perj´esi P. Naturally occurring chalcones and their biological activities. Phytochemistry Reviews, 2016, 15(1): 87-120. https://doi.org/10.1007/s11101-014-9387-8
  57. Tahir SK, Han EKH, Credo B, et al. A-204197, a new tubulin-binding agent with antimitotic activity in tumor cell lines resistant to known microtubule inhibitors. Cancer Research, 2001, 61(14): 5480-5485.
  58. Decordier I, Cundari E and Kirsch-Volders M. Survival of aneuploid, micronucleated and/or polyploid cells: Crosstalk between ploidy control and apoptosis. Mutation Research - Genetic Toxicology and Environmental Mutagenesis, 2008, 651(1-2): 30-39. https://doi.org/10.1016/j.mrgentox.2007.10.016
  59. Marrs KA. The functions and regulation of glutathione S-transferases in plants. Annual Review of Plant Physiology and Plant Molecular Biology, 1996, 47(1): 127-158. https://doi.org/10.1146/annurev.arplant.47.1.127
  60. Miyamoto T, Silva M and Hammock BD. Inhibition of epoxide hydrolases and glutathione Stransferases by 2-, 3-, and 4-substituted derivatives of 4’-phenylchalcone and its oxide. Archives of Biochemistry and Biophysics, 1987, 254(1): 203-213. https://doi.org/10.1016/0003-9861(87)90096-8
  61. Koleva YK, Madden JC and Cronin MTD. Formation of categories from structure-activity relationships to allow read-across for risk assessment: toxicity of α,β-unsaturated carbonyl compounds. Chemical Research in Toxicology, 2008, 21(12): 2300-2312. https://doi.org/10.1021/tx8002438
  62. Ivanova A, Batovska D, Engi H, et al. MDR-reversal activity of chalcones. In Vivo, 2008, 22(3): 379-384.
  63. Di Pietro A, Dayan G, Conseil G, et al. P-glycoprotein-mediated resistance to chemotherapy in cancer cells: Using recombinant cytosolic domains to establish structure-function relationships. Brazilian Journal of Medical and Biological Research, 1999, 32(8): 925-939. https://doi.org/10.1590/S0100-879X1999000800001
  64. Matsunaga H, Katano M, Saita T, et al. Potentiation of cytotoxicity of mitomycin C by a polyacetylenic alcohol, panaxytriol. Cancer Chemotherapy and Pharmacology, 1994, 33(4): 291-297. https://doi.org/10.1007/s002800050055
  65. Szliszka E, Czuba ZP, Mazur B, et al. Chalcones enhance TRAIL-induced apoptosis in prostate cancer cells. International Journal of Molecular Sciences, 2010, 11(1): 1-13. https://doi.org/10.3390/ijms11010001
  66. van der Oost R, Beyer J and Vermeulen NPE. Fish bioaccumulation and biomarkers in environmental risk assessment: a review. Environmental Toxicology and Pharmacology, 2003, 13(2): 57-149. https://doi.org/10.1016/S1382-6689(02)00126-6
  67. Arora A, Byrem TM, Nair MG, et al. Modulation of liposomal membrane fluidity by flavonoids and isoflavonoids. Archives of Biochemistry and Biophysics, 2000, 373(1): 102-109. https://doi.org/10.1006/abbi.1999.1525
  68. Kanojia D and Vaidya MM. 4-Nitroquinoline-1-oxide induced experimental oral carcinogenesis. Oral Oncology, 2006, 42(7): 655-667. https://doi.org/10.1016/j.oraloncology.2005.10.013
  69. Al-Qurainy F and Khan S. Mutagenic effects of sodium azide and its application in crop improvement. World Applied Sciences Journal, 2009, 6(12): 1589-1601. http://www.idosi.org/wasj/wasj6(12)/1.pdf
  70. Sadiq MF and Owais WM. Mutagenicity of sodium azide and its metabolite azidoalanine in Drosophila melanogaster. Mutation Research - Genetic Toxicology and Environmental Mutagenesis, 2000, 469(2): 253-257. https://doi.org/10.1016/S1383-5718(00)00079-6
  71. Torigoe T, Arisawa M, Itoh S, et al. Anti-mutagenic chalcones: antagonizing the mutagenicity of benzo(a)pyrene on Salmonella typhimurium. Biochemical and Biophysical Research Communications, 1983, 112(3): 833-842.
  72. Wang D, Lin Z,Wang T, et al. An analogous wood barrel theory to explain the occurrence of hormesis: A case study of sulfonamides and erythromycin on Escherichia coli growth. PLoS ONE, 2017, 12(7): e0181321. https://doi.org/10.1371/journal.pone.0181321
  73. Calabrese EJ. Biphasic dose responses in biology, toxicology and medicine: Accounting for their generalizability and quantitative features. Environmental Pollution, 2013, 182: 452-460. https://doi.org/10.1016/j.envpol.2013.07.046
  74. Ntuli SSBN, Gelderblom WCA and Katerere DR. The mutagenic and antimutagenic activity of Sutherlandia frutescens extracts and marker compounds. BMC Complementary and Alternative Medicine, 2018, 18(93): 1-10. https://doi.org/10.1186/s12906-018-2159-z
  75. Zeiger E. Mutagens that are not carcinogens: Faulty theory or faulty tests? Mutation Research - Genetic Toxicology and Environmental Mutagenesis, 2001, 492(1-2): 29-38. https://doi.org/10.1016/S1383-5718(01)00153-X
  76. Calabrese EJ. Hormesis: Principles and applications. Homeopathy, 2015, 104(2): 69-82. https://doi.org/10.1016/j.homp.2015.02.007
  77. Cushnie TPT and Lamb AJ. Antimicrobial activity of flavonoids. International Journal of Antimicrobial Agents, 2005, 26(5): 343-356. https://doi.org/10.1016/j.ijantimicag.2005.09.002
  78. Kirkland D, Reeve L, Gatehouse D, et al. A core in vitro genotoxicity battery comprising the Ames test plus the in vitro micronucleus test is sufficient to detect rodent carcinogens and in vivo genotoxins. Mutation Research - Genetic Toxicology and Environmental Mutagenesis, 2011, 721(1): 27-73. https://doi.org/10.1016/j.mrgentox.2010.12.015
  79. Nesslany F. The current limitations of in vitro genotoxicity testing and their relevance to the in vivo situation. Food and Chemical Toxicology, 2017, 106: 609-615. https://doi.org/10.1016/j.fct.2016.08.035