The spread of antibiotic-resistant bacteria in many humans and animals has driven researches to identify and design novel antibacterial agents. In vitro inhibitory activity of (2E)-2-(4,5-dihydro-4-oxothiazol-2-yl)-2-(thiazolidin-2-ylidene) acetonitrile against many bacterial pathogens has been proven in both veterinary and human medicine. In this study, its in vivo toxic effects was studied in mice. The median lethal dose (LD50) value of 239.88 mg/kg was estimated using intraperitoneal injection in 8 groups of mice after 48 h treatment. Then, intraperitoneal injections of LD50 of oxothiazole solution into 4 other mice were done to evaluate histopathological changes in their liver and kidney tissues. The histopathological studies were identified as fatty change, hepatitis, necrosis and regeneration in liver, and fibrosis, necrosis, nephritis, hyaline cast and hyperaemia in kidney. In conclusion, the synthesized oxothiazole derivative causes renal and hepatic toxicity in mice at medium concentrations. The change of thiazole substituents and complexation may reduce its toxicity

Abbas Jamshidian

Behzad Ghasemi

Fatemeh Arab-Sahebi

Hamid Beyzaei

Maliheh Abdollahi

Mohammad Reza Hajinezhad

Nasim Amel

Seyed Hadi Hashemi

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Corresponding author: Behzad GhasemiTell/Fax: +98 54 31232186Email: behzad.ghasemi99@gmail.comIranian Journal of VeterinaryScience and Technology Received:  Accepted after revision: Published online: DOI:  10.22067/veterinary.v2i10.71626RESEARCH ARTICLEABSTRACTKeywords Abbreviationswww.IJVST.um.ac.ir The spread of antibiotic-resistant bacteria in many humans and animals has driven researches to identify and design novel antibacterial agents. In vitro inhibitory activity of (2E)-2-(4,5-di-hydro-4-oxothiazol-2-yl)-2-(thiazolidin-2-ylidene) acetonitrile against many bacterial pathogens has been proven in both veterinary and human medicine. In this study, its in vivo toxic effects was studied in mice. The median lethal dose (LD50) value of 239.88 mg/kg was estimated using intraperitoneal injection in 8 groups of mice after 48 h treatment. Then, intraperitoneal injections of LD50 of oxothiazole solution into 4 other mice were done to evaluate histopathological chang-es in their liver and kidney tissues. The histopathological studies were identified as fatty change, hepatitis, necrosis and regeneration in liver, and fibrosis, necrosis, nephritis, hyaline cast and hy-peraemia in kidney. In conclusion, the synthesized oxothiazole derivative causes renal and hepatic toxicity in mice at medium concentrations. The change of thiazole substituents and complexation may reduce its toxicityStudy of Toxic Effects of Oxothiazole Derivative as a New Antibacterial AgentHamid Beyzaei, Abbas Jamshidian, Mohammad Reza Hajinezhad, Seyed Hadi Hashemi, Nasim Amel , Maliheh Abdollahi, Fatemeh Arab-Sahebi, Behzad Ghasemi,          b          c         a        daDepartment of Chemistry, Faculty of Science, University of Zabol, Zabol, Iran. bDepartment of Pathobiology, Faculty of Veterinary Medicine, University of Zabol, Zabol, Iran. cDepartment of Basic Sciences, Faculty of Veterinary Medicine, University of Zabol, Zabol, Iran. dDepartment of Clinical Sciences, Faculty of Veterinary Medicine, Zabol University, Zabol, Iran. eStudent Research Committee, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh, Iran. fTorbat Jam Faculty of Medical Sciences, Torbat Jam, Iran. gStudent Research Committee, Sabzevar University of Medical Sciences, Sabzevar, Iran.Toxic effects, Histopathological study, Oxothiazole de-rivativeLD50: Median Lethal Dose       f        g        f       e2018-Mar-122019- Mar- 102018-Oct-12RESEARCH ARTICLEIJVST 2018-2 (19) DOI:10.22067/veterinary.v2i10.7162618Introduction    Species of pathogenic bacteria have developed resistance to current antibiotics such as peni-cillin, amoxicillin, clindamycin, tetracycline, erythro-mycin and sulfamethoxazole due to overuse of drugs in humans and animals [1]. This health problem in-creases costs of medical care [2]. In recent years, many researchers were encouraged to discover new antibac-terial agents including herbal extracts, metal nanopar-ticles and heterocyclic derivatives [3]. Thiazoles are a major class of heterocyclic com-pounds, which have been used in the treatment of dis-eases such as cancer, hypertension and lipid disorders [4]. Significant antibacterial effects were observed by thiazole derivatives. It was found that they can inhibit the activity of DNA gyrase B and ecKAS III enzymes, and were effective against the replication of DNA and the synthesis of fatty acids, in bacteria [5].In vitro inhibitory activities of thiazoles have been assessed against a variety of pathogenic bacte-ria including both Gram-positive and Gram-negative strains [6-8]. These compounds inhibit the growth of bacteria by blocking ion channels as well as inhibit-ing enzymatic and cellular activities [9-12]. However, there are a few in vivo studies on body and organ of animals.Oxothiazole derivative is a recently synthesized heterocyclic compound that its in vitro inhibitory ac-tivities were evaluated against some Gram-positive and Gram-negative bacteria such as Staphylococcus au-reus, Streptococcus agalactiae, Listeria monocytogenes, Bacillus cereus, Escherichia coli and Salmonella typh-imurium [13-16]. The present study was performed to assess the function and histology of the liver and kidney in mice exposed to the synthetic oxothiazole. Results   The toxic and therapeutic effects of oxothiazole derivative were evaluated in 8 groups of 8 mice each. The 48 h LD50 value of compound was 239.88 mg/kg (Tables 1 and 2, Figure 1). Histopathological changes including fatty change, hepatitis, necrosis and regen-Table 1 Mortality percent in each group after IP injection of oxothi-azole.TotalLivedPercentDiedlogDose mg/kg88002.201608712.512.30200862522.34220845042.38240827562.422658010082.473008010082.54350Table 2Different LD doses of  oxotiazole190.55LD10230.14LD40239.88LD50267.92LD80278.61LD90Figure 1Mortality of mice infect-ed with oxothiazoleeration in liver, and fibrosis, necrosis, nephritis, hya-line cast and hyperaemia in kidney were observed as illustrated in Figures 2 and 3.Discussion   Thiazole ring is an essential part of numerous bi-ologically active compounds. Thiazole derivatives are recognized to affect cell cycle progression, protein production and DNA replication [17-19]. Sulfathi-azole as an antibiotic containing thiazole ring was used in the treatment of gastrointestinal and respira-tory infections. Its oral LD50 in mice is 4500 mg/kg. Although toxicity of the drug is low, it is prescribed less frequently in humans due to its side effects such as high fever, severe headache, stomatitis, conjunctivi-19IJVST DOI: 10.22067/veterinary.v2i10.716262018-2 (19)RESEARCH ARTICLEtis, rhinitis, urethritis and balanitis [20]. Febuxostat is a xanthine oxidase inhibitor containing thiazole ring that lowers uric acid levels in blood. The LD50 of drug administered orally to mice is 300 mg/kg [21]. Spry-cel (dasatinib) is a chemotherapy medication used to treat chronic myelogenous leukemia (CML) and acute lymphoblastic leukemia (ALL). It has a relatively high toxicity (oral LD50 = 100 mg/kg) in rat. Its consump-tion may cause anemia, swelling, pulmonary edema, heart failure, rash and diarrhea [22].Anticoagulant and antiarrhythmic effects of some thiazole derivatives were studied by Abd El-Galil et al. in 2009 via determination of LD50 values of 292-736 mg/kg. Although oxothiazole is more toxic than these derivatives, the determination of median effec-tive dose (ED50) and therapeutic index (TI) of this heterocycle in future researches is necessary for a bet-ter conclusion [23]. Siddiqui et al. in 2010 calculated the toxic effects of 3-[4-(substituted phenyl)-1,3-thi-azol-2-ylamino]-4-(substituted phenyl)-4,5-dihydro-1H-1,2,4-triazole-5-thiones as new anticonvulsant agents, their LD50 values (834 and 936 mg/mL) are higher than those in phenytoin (a common anticon-vulsant drug with LD50 = 224 mg/mL) [24].Efficiency of some synthesized imidazo[2,1-b]thiazole derivatives as diuretic agents was proven in comparison with those of hydrochlorothiazide and furosemide, LD50 values were reported in the range of 1000 to 1500 mg/mL [25].Among 12 thiazolidines synthesized by Wang et al., only three derivatives showed significant an-ti-inflammatory activities. They can be used to treat Figure 2Histopathological changes of liver tissue in mice treated with oxothiazoleFigure 3 Histopathological changes of kidney tissue in mice treated with oxothiazolehepatic injuries according to the results obtained by further Con A-induced acute liver injury of BALB/c mice [26]. Thiazole-Zn is a chinese-created systemic fungicide that had no pathological effects on the liver and kidney of female rats when used as oral gavage, while its destructive effects were observed in the thy-roid gland [27].The toxicity of oxothiazole derivative has been preliminarily studied in mice at doses of 350 and 160 mg/kg without determination of LD50. All mice died at dose of 350 mg/kg after 24 h. The liver of mice treat-ed at dose 160 mg/kg was evaluated after 48 h [15]. To extend toxicological studies, some biochemical and physiological effects of this potential antibacterial agent were assessed via determination of LD50 value and evaluation of histopathological changes in the liv-er and kidney of mice. Renal and hepatic toxicity were observed in mice when medium doses of heterocyclic derivative were injected. New oxothiazole derivatives with low toxicity can be designed in future studies by changing thiazole substituents and complexation. Material and methods   General procedure for the synthesis of oxothi-azole compoundOxothiazole (3) was synthesized according to a previous procedure [28] as follows: a suspension of 1 mmol of each (E)-2-cyano-2-(thiazolidin-2-ylidene) ethanethioamide (1), ethyl bro-moacetate (2) and sodium bicarbonate (NaHCO3) in 1 mL DMF as solvent was stirred at room temperature for 8 h. 5 mL of water was added to the reaction mixture. The precipitate was filtered out, and washed with water (1 × 5 mL) and ethanol (1 × 5 mL), RESEARCH ARTICLEIJVST 2018-2 (19) DOI:10.22067/veterinary.v2i10.7162620respectively. The pure thiazole 3 was obtained in 75% yield after recrystallization from acetonitrile, and its chemical structure was confirmed through the elemental analysis and spectroscopic data (Figure 4).m.p. 248-252 °C; IR (KBr) ν: 3422 (NH), 2193 (C≡N), 1688 (C=O), 1573 (C=C) cm-1; 1H NMR (DMSO-d6) δ: 3.63 (t, J = 7.8 Hz, 2H, SCH2), 4.01 (s, 2H, COCH2), 4.12 (t, J = 7.8 Hz, 2H, NHCH2), 10.53 (s, 1H, NH) ppm; MS (EI, m/z): 225 (M+, 21), 176 (37), 150 (94), 109 (28), 46 (100); Anal. Calcd for C8H7N3OS2: C, 42.65; H, 3.13; N, 18.65. Found: C, 42.74; H, 3.00; N, 18.46.In vivo study68 white male BALB/c mice, weighing 26-34 g, were obtained from the Central Animal House, University of Zabol, Zabol, Iran. Mice were maintained at all stages with free access to standard food and refined water under 25 ± 2 oC temperature, 50% humid-ity, 12/12 light dark cycle [24].Assessment of median lethal dose (LD50)An up-and-down method was applied for acute toxicity testing. 64 mice were divided into 8 groups. Finally, doses includ-ing 160, 200, 220, 240, 265, 300 and 350 mg/kg of oxothiazole in 10% DMSO were selected for intraperitoneal injection (IP). 10% DMSO was also injected as negative control. Mice mortality was recorded in each group during 48 h [24]. Histopathological examination4 BALB/c were injected intraperitoneally with the median lethal doses of oxothiazole. The abdominal walls of dead and live mice were excised immediately or after ether euthanasia. The liver and kidney of mice were removed from their abdominal cavity, and stored in 10% formalin. All solutions were refreshed three times every 24 h. The histological sections were molded in 4- to 6-μm-thick paraffin according to the histopathological principles. The prepared slides were stained with hematoxylin and eosin, and examined under a light microscope [27].Acknowledgment   This work was supported by the University of Zabol under Grant number UOZ-GR-9517-15.Author Contributions   All authors contributed to the design of study, data analysis and manuscript preparation.Conflict of Interest   The authors declare that there is no conflict of in-terest.References   1. Wang D, Zhang L, Zhou X, He Y, Yong C, Shen M, et al. An-timicrobial susceptibility, virulence genes, and randomly am-plified polymorphic DNA analysis of Staphylococcus aureus recovered from bovine mastitis in Ningxia, China.J Dairy Sci. 2016;99(12):9560-9.2. Ahumada-Santos YP,  Soto-Sotomayor ME, Baez-Flores ME, Diaz-Camacho SP, Lopez-Angulo G , Eslava-Campos CA, et al. Antibacterial synergism of Echeveria subrigida (B. L. Rob & Seaton) and commercial antibiotics against multidrug re-sistant Escherichia coli and Staphylococcus aureus.Eur J Int Med. 2016;8(5):638-44.3. Santos L, Ramos F. Analytical strategies for the detection and quantification of antibiotic residues in aquaculture fishes: A review. Trends Food Sci Technol. 2016;52:16-30.4. Bakavoli M, Beyzaei H, Rahimizadeh M, Eshghi H. Regiose-lective synthesis of 2[(E)(benzo[d]thiazol2(3H)ylidene)(cya-no)methyl]thiazoles. Heterocycl Commun. 2011;17:151-4.5. Ghasemi B, Sanjarani G, Sanjarani Z, Majidiani H. Evaluation of anti-bacterial effects of some novel thiazole and imidazole derivatives against some pathogenic bacteria. Iran J Microb. 2015;7(5):281-6.6. Beyzaei H, Aryan R, Molashahi H, Zahedi MM, Samza-deh-Kermani A Ghasemi B. MgO nanoparticle-catalyzed, sol-vent-free Hantzsch synthesis and antibacterial evaluation of new substituted thiazoles. J Iran Chem Soc. 2017;14(5):1023-31.7. Pawar CD, Sarkate AP, Karnik KS, Bahekar SS, Pansare DN, Shelke RN, et al. Synthesis and antimicrobial evaluation of novel ethyl 2-(2-(4-substituted)acetamido)-4-subtituted-thi-azole-5-carboxylate derivatives. Bioorg Med Chem Lett. 2016;26(15):3525-8.8. Qin YJ, Wang PF, Makawana JA, Wang ZC, Wang ZN,Gu Y,et al. Design, synthesis and biological evaluation of metronida-zole–thiazole derivatives as antibacterial inhibitors. Bioorg Med Chem Lett. 2014;24(22):5279-83.9. Washburn DG, Holt DA, Dodson J, McAtee JJ, Terrell LR, Barton L, et al. The discovery of potent blockers of the canon-ical transient receptor channels, TRPC3 and TRPC6, based on an anilino-thiazole pharmacophore. Bioorg Med Chem Lett. 2013;23:4979-84.10. Kang SY, Song KS, Lee J, Lee SH, Lee J. Synthesis of pyri-dazine and thiazole analogs as SGLT2 inhibitors. Bioorg Med Chem Lett. 2010;18:6069-79.11. Andreani A, Granaiola M, Guardigli M, Leoni A, Locatel-li A, Morigi R, et al. Synthesis and chemiluminescent high throughput screening for inhibitionof acetylcholinesterase activity by imidazo[2,1-b]thiazole derivatives. Eur J Med Chem. 2005;40:1331-4.12. Andreani A, Granaiola M, Leoni A, Locatelli A, Morigi R, Rambaldi M. Effects of new ubiquinone-imidazo[2,1-b]thi-azoles on mitochondrial complex I (NADH-ubiquinone re-ductase) and on mitochondrial permeability transition pore. Bioorg Med Chem Lett. 2004;12:5525-32.SHNCNSNH2EtOOBrSHNCNSNO   DMF, NaHCO3+1                                2                                                     3Figure 4 Synthesis of oxothiazole derivative 3(2E)-2-(4,5-Dihydro-4-oxothiazol-2-yl)-2-(thiazolidin-2-ylidene)acetonitrile.21IJVST DOI:10.22067/veterinary.v2i10.716262018-2 (19)RESEARCH ARTICLE13. Ghasemi B, Beyzaei H, Hashemi SH, Majidiani H. Study of antibacterial effect ofnovel thiazole, imidazole and tetrahydropyridine derivatives against Escherichia coli. J Med Bacteriol. 2015;4(6):7-12.14. Ghasemi B, Beyzaei H, Hashemi SH. Study of antibacterial effect of novel thiazole, imidazole, and tetrahydro-pyrimidine derivatives against Listeria Monocytogens. Ann Mil Health Sci Res. 2015;13:101-5.15. Ghasemi B, Najimi M, Beyzaei H, Jamshidian A. Antibacterial effects of novel thiazole derivatives and the toxicity of oxothiazole on liver histopathology of mice. Med Lab J. 2015;9(4):6-12.16. Ghasemi B , Beyzaie H , Majidiani H.A comparative study on the anti-bacterial effects of some newly synthe-sized thiazole, imidazolidine and tet-rahydropyrimidine derivatives against Bacillus cereus and Salmonella typh-imurium. Pharm Sci. 2016;22(1):54-9.17. Chen H, Song J, Wang J, Xu C, Chen C, Gu W. Synthesis and biological evaluation of thiazole derivatives as novel USP7 inhibitors. Bioorg Med Chem Lett. 2017;27(4):845-9.18. Reichelt A, Bailis JM, Bartberger MD, Yao G, Shu H, Kaller MR.  Synthesis and structure-activity relationship of trisubstituted thiazoles as Cdc7 kinase inhibitors. Eur J Med Chem. 2014;80:364-82. 19. Chen YL, Cherry K, Michael L, Ebb-inghaus CF,  Gamlath CB, Liston D. Thiazole-diamides as potent γ-secre-tase inhibitors. Bioorg Med Chem Lett. 2007;17(20):5518-22.20. Prinsen MK, Romijn T, Snoeij NJ. Skin sensitization testing: the relevance of rechallenge and pretreatment with so-dium lauryl sulfate in the guinea pig maximization test. Food Chem Toxi-col. 1997;35(9):923-6.21. Love BL, Barrons R, Veverka A, Snider KM. Urate-lowering therapy for gout: focus on febuxostat. Pharmacother. 2010;30(6):594-608.22. Keating, GM. Dasatinib: A review in chronic myeloid leukaemia and Ph+ acute lymphoblastic leukaemia. Drugs. 2017;77(1):85-9.23. Abd El-Galil EA, Sabrry NM, Abdalla MM, Abdel-Wahab BF. Synthesis, an-tiarrhythmic and anticoagulant activi-ties of novel thiazolo derivatives from methyl 2-(thiazol-2-ylcarbamoyl)ace-tate. Eur J Med Chem. 2009;44:725-35.24. Siddiqui N, Ahsan W. Triazole in-corporated thiazoles as a new class of anticonvulsants: Design, synthe-sis and in vivo screening. Eur J Med Chem.2010;45:1536-43.25. Andreani A, Rambaldi M, Mascellani G, Rugarli P. Synthesis and diuret-ic activity of imidazo[2,1-b]thiazole acetohydrazones. Eur J Med Chem. 1987;22(1):19-22.26. Wang G, Deng C, Xie C, Ma L, Yang J, QiuN, et al. Synthesis and biological evaluation of novel dimethyl[1,10-bi-phenyl]-2,20-dicarboxylate derivatives containing thiazolidine-2,4-dione for the treatment of concanavalin A-in-duced acute liver injury of BALB/c mice. Eur J Med Chem. 2011;46:5941-8.27. Yang H, Zhang W, Kong Q, Liu H, Sun R, Lin B, et al. Effects of pubertal expo-sure to thiazole-Zn on thyroid func-tion and development in female rats. Food Chem Toxicol.2013;53:100-4.28. Bakavoli M, Beyzaei H, Rahimizadeh M, Eshghi H, Takjoo R. Regioselective synthesis of new 2-(E)-cyano(thiazo-lidin-2-ylidene)thiazoles. Molecules. 2009;14:4849-57.

Study of Toxic Effects of Oxothiazole Derivative as a New Antibacterial Agent

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Study of Toxic Effects of Oxothiazole Derivative as a New Antibacterial Agent

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