The treatment of multidrug-resistant tuberculosis (MDR-TB) is a challenge to be overcome. The increase of resistant isolates associated with serious side effects during therapy leads to the search for substances that have anti-TB activity, which make treatment less toxic, and also act in the macrophage acidic environment promoted by the infection.
Objective:
The aim of this study was to investigate lapachol and β-lapachone activities in combination with other drugs against Mycobacterium tuberculosis at neutral and acidic pH and its cytotoxicity.
Design:
Inhibitory and bactericidal activities against M. tuberculosis and clinical isolates were determined. Drug combination and cytotoxicity assay were carried out using standard TB drugs and/or N-acetylcysteine (NAC).
Results:
Both naphthoquinones presented activity against MDR clinical isolates. The combinations with the first-line TB drugs demonstrated an additive effect and β-lapachone+NAC were synergic against H37Rv. Lapachol activity at acidic pH and its association with NAC improved the selectivity index. Lapachol and β-lapachone produced cell morphological changes in bacilli at pH 6.0 and 6.8, respectively.
Conclusion:
Lapachol revealed promising anti-TB activity, especially associated with NAC.
Ministry of Health. 2011. Manual of recommendations for tuberculosis control in Brazil. Ministry of Health, Health Surveillance, Secretary, Brasilia.
4.
ZhangY., and YewW.. 2015. Mechanisms of drug resistance in Mycobacterium tuberculosis: update. Int. J. Lung. Dis. 19:1276–1289.
5.
Mitchison, D.A. 1985. The action of antituberculosis drugs in short-course chemotherapy. Tubercle, 66:219–225.
6.
VandalO.H., NathanC.F, and EhrtS.. 2009. Acid resistance in Mycobacterium tuberculosis. J. Bacteriol. 191:4714–4721.
7.
ZhangY., PermarS., and SunZ.. 2002. Conditions that may affect the results of susceptibility testing of Mycobacterium tuberculosis to pyrazinamide. J. Med. Microbiol. 51:42–49.
8.
EyongK.O., KumarS.P, KueteV., FolecfocN., LangmiH., Marion LallN., and BaskaranS.. 2012. Cobalt mediated ring contraction reaction of lapachol and initial antibacterial evaluation of naphthoquinones derived from lapachol. Med. Chem. Res. 21:2117–2122.
9.
da SilvaJ.L., MesquitaA.R.C, and XimenesE.A.. 2009. In vitro synergic effect of β-lapachone and isoniazid on the growth of Mycobacterium fortuitum and Mycobacterium smegmatis. Mem. Inst. Oswaldo Cruz. 104:580–582.
10.
PinkJ.J., PlanchonS.M, TagliarinoC., VarnesM.E, SiegelD., and BoothmanD.A.. 2000. NAD(P)H:Quinone oxidoreductase activity is the principal determinant of β-lapachone cytotoxicity. J. Biol. Chem. 275:5416–5424.
11.
SiegelD., YanC., and RossD.. 2012. NAD(P)H:quinone oxidoreductase 1 (NQO1) in the sensitivity and resistance to antitumor quinones. Biochem. Pharmacol. 83:1033–1040.
12.
CossarizzaA., FranceschiC., MontiD., SalvioliS., BellesiaE., RivabeneR., BionoL., RainaldiG., TinariA., and MalorniW.. 1995. Protective effect of N-acetylcysteine in tumor necrosis factor-α-induced apoptosis in U937 cells: the role of mitochondria. Exp. Cell. Res. 220:232–240.
13.
BhusalY., ShiohiraC.M, and YamaneN.. 2005. Determination of in vitro synergy when three antimicrobial agents are combined against Mycobacterium tuberculosis; Int. J. Antimicrob. Agents. 26:292–297.
14.
GarcíaA., Bocanegra-GarcíaV., Palma-NicolásJ.P, and RiveraG.. 2012. Recent advances in antitubercular natural products. Eur. J. Med. Chem. 49:1–23.
15.
Ferreira, V.F. 1996. Learning about the concepts of acid and base. Quim. Nova. na Esc. 4:35–6.
16.
BarbosaT.P., and NetoH.D.. 2012. Preparation of lapachol derivatives in acid and basic media: an experiment proposal for the discipline of experimental organic chemistry. Quim. Nova. 36:1540–1551.
17.
PalominoJ., MartinA., CamachoM., GuerraH., SwingsJ., and PortaelsF.. 2002. Resazurin microtiter assay plate: simple and inexpensive method for detection of drug resistance in Mycobacterium tuberculosis. Antimicrobial Agents Chemother. 46:2720–2722.
Caleffi-FerracioliK.R., MaltempeF.G, SiqueiraV.L.D, and CardosoR.F.. 2013. Fast detection of drug interaction in Mycobacterium tuberculosis by a checkerboard resazurin method. Tuberculosis, 93:660–663.
20.
Odds, F.C. 2003. Synergy, antagonism, and what the chequerboard puts between them. J. Antimicrob. Chemother. 52:1.
21.
PiresC.T.A., BrenzanM.A, ScodroR.B.L, CortezD.A, LopesL.D, SiqueiraV.L.D, and CardosoR.F.. 2014. Anti-Mycobacterium tuberculosis activity and cytotoxicity of Calophyllum brasiliense Cambess (Clusiaceae), Memórias do Inst. Oswaldo Cruz. 109:324–329.
22.
ZhangY., WadeM.M, ScorpioA., ZhangH., and SunZ.. 2003. Mode of action of pyrazinamide: disruption of Mycobacterium tuberculosis membrane transport and energetics by pyrazinoic acid. J. Antimicrob. Chemother. 52:790–795.
23.
ZhangL.Y., and AmzelL.M.. 2002. Tuberculosis drug targets. Curr. Drug Targets, 3:131–154.
24.
CarneiroP.F., M.D.PintoC.F.R, CoelhoT.S, CavalcantiB.C, PessoaC., de SimoneC.A, NunesI.K, de OliveiraN.M, de AlmeidaR.G, PintoA.V, de MouraK.C, da SilvaP.A, and da Silva JúniorE.N.. 2011. Quinonoid and phenazine compounds: synthesis and evaluation against H37Rv, rifampicin and isoniazid-resistance strains of Mycobacterium tuberculosis, Eur. J. Med. Chem. 46:4521–4529.
25.
OliveiraR.A.S., Azevedo-XimenesE., LuzzatiR., and GarciaR.C.. 2010. The hydroxy-naphthoquinone lapachol arrests mycobacterial growth and immunomodulates host macrophages. Int. Immunopharmacol. 10:1463–1473.
26.
SilvaM.N., FerreiraV.F, and M.SouzaC.B.V. 2003. An overview of the chemistry and pharmacology of naphthoquinones with emphasis on β-lapachone and derivatives. Quím. Nova. 26:407–416.
27.
DelaraminaM., NicolettiC.D, MoraesM.C, FuturoD.O, BühlM., SilvaF.C, FerreiraV.F, an Md CarneiroJ.W. 2019. α- and β-lapachone isomerization in acid media: insights from experimental and implicit/explicit solvation approaches. Chem. Eur. 84:52–61.
28.
CoelhoT.S., SilvaR.S.F, PintoA.V, PintoM.C, ScainiC.J, MouraK.C, and da SilvaA.P.. 2010. Activity of β-lapachone derivatives against rifampicin-susceptible and -resistant strains of Mycobacterium tuberculosis.Tuberculosis. 90:293–297.
29.
PankeyG. A., and SabathL.D.. 2018. Clinical relevance of bacteriostatic versus bactericidal mechanisms of action in the treatment of Gram-positive bacterial infections. Clin. Infect. Dis. 38:864–870.
30.
NemethJ., OeschG., and KusterS.P.. 2018. Bacteriostatic versus bactericidal antibiotics for patients with serious bacterial infections: systematic review and meta-analysis. J. Antimicrob. Chemother. 70:382–395.
31.
JainS.N., VishwanathaT., ReenaV., DivyashreeB.C, AishwaryaS., SiddhalingeshwarK.G, VenugopalN., and RameshI.. 2011. Antibiotic synergy test: checkerboard method on multi drug resistant Pseudomonas aeruginosa. Int. Res. J. Pharm. 2:196–198.
32.
AmaralE.P., ConceiçãoE.L, CostaD.L, RochaM.S, MarinhoJ.M, Cordeiro-SantosM., D'Império-LimaM.R, BarbosaT., SherA., and AndradeB.B.. 2016. N-acetylcysteine exhibits potent anti-mycobacterial activity in addition to its known anti-oxidative functions. Biomed. Cent. Microbiol. 16:251.
33.
BaniasadiS., EftekhariP., TabarsiP., FahimiF., RaoufyM.R, MasjediM.R, and VelayatiA.A.. 2010. Protective effect of N-acetylcysteine on antituberculosis drug-induced hepatotoxicity. Eur. J. Gastroenterol. Hepatol. 22:1235–1238.
34.
KranzerK., ElaminW.F, CoxH., SeddonJ.A, FordN., and DrobniewskiF.. 2015. A systematic review and meta-analysis of the efficacy and safety of N-acetylcysteine in preventing aminoglycoside-induced ototoxicity: implications for the treatment of multidrug-resistant TB. Thorax, 70:1070–1071.
JuhaszA.L., SmithE., NelsonC., ThomasD.J, and BradhamK.. 2014. Variability associated with as in vivo-in vitro correlations when using different bioaccessibility methodologies. Environ. Sci. Technol. 48:11646–11653.
37.
SunasseeS.N., VealeC.G.L, Shunmoogam-GoundenN., OsoniyiO., HendricksD.T, CairaM.R, de la MareJ.A, EdkinsA.L, PintoA.V, da Silva JúniorE.N, and Davies-ColemanM.T.. 2013. Cytotoxicity of lapachol, β-lapachone and related synthetic 1,4-naphthoquinones against oesophageal cancer cells. Eur. J. Med. Chem. 62:98–110.
38.
PereiraE.M., MachadoT.D.B, LealI.C, JesusD.M, DamasoC.R, PintoA.V, Giambiagi-de MarvalM., KusterR.M, and dos SantosK.R.N. 2006. Tabebuia avellanedae naphthoquinones: activity against methicillin-resistant staphylococcal strains, cytotoxic activity and in vivo dermal irritability analysis. Ann. Clin. Microbiol. Antimicrob. 5:1–7.
39.
CostaE.V.S., BrígidoC.H.P, J.V.daS, Silva, Coelho-FerreiraM.R, BrandãoG.C, and DolabelaM.F.. 2017. Antileishmanial activity of Handroanthus serratifolius (Vahl) S. Grose (Bignoniaceae). Evid. Based Complement. Altern. Med. 2:1–6.
40.
OrmeI.2001. Search for new drugs for treatment of tuberculosis. Antimicrobial Agents Chemother. 45:1943–1946.
41.
RochaM.N., NogueiraP.M, DemicheliC., OliveiraL.G, SilvaM.M, FrézardF., MeloM.N, and SoaresR.P.. 2013. Cytotoxicity and in vitro antileishmanial activity of antimony (V), bismuth (V), and tin (IV) complexes of lapachol. Bioinorg. Chem. Appl. 2013:961783.
42.
TabriziL., TalaieF., and ChiniforoshanH.. 2017. Copper(II), cobalt(II) and nickel(II) complexes of lapachol: synthesis, DNA interaction, and cytotoxicity. J. Biomol. Struct. Dyn. 35:3330–3341.
43.
SalustianoE.J.S., NettoC.D, FernandesR.F, da SilvaA.J, BacelarT.S, CastroC.P, BuarqueC.D, MaiaR.C, RumjanekV.M, and CostaP.R.. 2010. Comparison of the cytotoxic effect of lapachol, β-lapachone and pentacyclic 1,4-naphthoquinones on human leukemic cells. Invest. New Drugs, 28:139–144.
44.
Kerksick, C. and WilloughbyD.. 2005. The antioxidant role of glutathione and n-acetyl-cysteine supplements and exercise-induced oxidative stress. J. Int. Soc. Sports Nutr. 2:38–44.
45.
SunX., LiY., LiW., ZhangB., WangA.J, SunJ., MikuleK., JiangZ., and LiC.J.. 2006. Selective induction of necrotic cell death in cancer cells by β-lapachone through activation of DNA damage response pathway. Cell Cycle. 5:2029–2035.
46.
MaX., HuangX., HuangG., LiL., WangY., LuoX., BoothmanD.A, and GaoJ.. 2014. Prodrug strategy to achieve lyophilizable, high drug loading micelle formulations through diester derivatives of β-lapachone. Adv. Healthc. Mater. 3:1210–1216.
47.
KungH.N., WengT.Y, LiuY.L, LuK., Sand ChauY.P. 2014. Sulindac compounds facilitate the cytotoxicity of β-lapachone by up-regulation of NAD(P)H quinone oxidoreductase in human lung cancer cells. PLoS One, 9:e88122.
48.
da Silva JúniorE.N., de DeusC.F, CavalcantiB.C, PessoaC., Costa-LotufoL.V, MontenegroR.C, de MoraesM.O, M.d.PintoC., de SimoneC.A, FerreiraV.F, GoulartM.O, AndradeC.K, and PintoA.V.. 2010. 3-arylamino and 3-alkoxy-nor-β-lapachone derivatives: synthesis and cytotoxicity against cancer cell lines. J. Med. Chem. 53:504–508.
49.
FlickH.E., LaLondeJ.M, MalachowskiW.P, and MullerA.J.. 2013. The tumor-selective cytotoxic agent β-lapachone is a potent inhibitor of IDO1. Int. J. Tryptophan Res. 6:35–45.
50.
PaludoC.R., da Silva-JuniorE.A, SilvaE.O, VessecchiR., LopesN.P, PupoM.T, F.da EmeryS., dos SN.. Gonçalves, dos SantosR.A, and N.FurtadoA.J.C. 2017. Inactivation of β-lapachone cytotoxicity by filamentous fungi that mimic the human blood metabolism. Eur. J. Drug Metab. Pharmacokinet. 42:213–220.
51.
Silva EO., de CarvalhoT.C, ParshikovI.A, SantosR.A, EmeryS.F, and N.FurtadoA.J.C. 2014. Cytotoxicity of lapachol metabolites produced by probiotics, Lett. Appl. Microbiol. 59:108–114.
52.
LeeJ.I., ChoiD.Y, ChungH.S, SeoH.G, WooH.J, ChoiB.T, and ChoiY.H.. 2006. β-lapachone induces growth inhibition and apoptosis in bladder cancer cells by modulation of Bcl-2 family and activation of caspases. Exp. Oncol. 28:30–35.
53.
LiuT.J., LinS.Y, and ChauY.P.. 2002. Inhibition of poly(ADP-ribose) polymerase activation attenuates β-lapachone-induced necrotic cell death in human osteosarcoma cells. Toxicol. Appl. Pharmacol. 182:116–125.
54.
ReinickeK.E., BeyE.A, BentleM.S, PinkJ.J, IngallsS.T, HoppelC.L, MisicoR.I, ArzacG.M, BurtonG., BornmannW.G, SuttonD., GaoJ., and BoothmanD.A.. 2005. Development of β-lapachone prodrugs for therapy against human cancer cells with elevated NAD(P)H:Quinone oxidoreductase 1 levels. Clin. Cancer Res. 11:3055–3064.
55.
RauthA., GoldbergZ., and MisraV.. 1997. DT-diaphorase: possible roles in cancer chemotherapy and carcinogenesis. Oncol. Res. 9:339–349.
56.
Campanerut-SáP.A.Z., Ghiraldi-LopesL.D, MeneguelloJ.E, FioriniA., EvaristoG.P, SiqueiraV.L.D, ScodroR.B.L, PatussiE.V, DonattiL., SouzaE.M, and CardosoR.F.. 2016. Proteomic and morphological changes produced by subinhibitory concentration of isoniazid in Mycobacterium tuberculosis.Future Microbiol. 11:1123–1132.
57.
Ghiraldi-LopesL.D., Campanerut-SáP.A.Z, MeneguelloJ.E, SeixasF.A, Lopes-OrtizM.A, ScodroR.B.L, PiresC.T, da SilvaR.Z, SiqueiraV.L.D, NakamuraS.C.V, and CardosoR.F.. 2017. Proteomic profile of Mycobacterium tuberculosis after eupomatenoid-5 induction reveals potential drug targets. Future Microbiol. 12:867–879.
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