Arrabidaea harleyi A.H. Gentry (Bignoniaceae) is an ornamental plant found in some regions of the Atlantic forest in Brazil. From its bark a mixture of verbascoside and isoverbascoside was isolated. This mixture was shown to be active againstStaphylococcus aureus, Micrococcus luteus, Bacillus subtilis, Bacillus mycoides, Enterecoccus faecalis, Escherichia coli, Serratia marcensisand Candida albicans. The minimal inhibitory concentration (MIC) was established by diffusion method.Arrabidaea harleyi A.H. Gentry (Bignoniaceae) é uma planta ornamental, encontrada em algumas regiões da Mata Atlântica do Brasil. A partir das cascas do caule foi isolada a mistura dos isômeros verbascosídeo e isoverbascosídeo. A mistura mostrou-se ativa frente a Staphylococcus aureus, Micrococcus luteus, Bacillus subtilis, Bacillus mycoides, Enterococcus faecalis, Escherichia coli, Serratia marcensis e Candida albicans. Foi estabelecida a concentração mínima inibitória (CMI) através do método de difusão em meio sólido

Agra, Maria de Fátima

Amorim, Elba Lúcia Cavalcanti de

Barbosa-Filho, José Maria

Chiappeta, Alda de Andrade

da-Cunha, Emidio Vasconcelos Leitão

Lima, Cláudia Sampaio de Andrade

Nunes, Xirley Pereira

Sena, Kêsia Xisto da Fonseca Ribeiro de

Silva, Marcelo Sobral da

English

Arrabidaea harleyi A.H. Gentry (Bignoniaceae) é uma planta ornamental, encontrada em algumas regiões da Mata Atlântica do Brasil. A partir das cascas do caule foi isolada a mistura dos isômeros verbascosídeo e isoverbascosídeo. A mistura mostrou-se ativa frente a Staphylococcus aureus, Micrococcus luteus, Bacillus subtilis, Bacillus mycoides, Enterococcus faecalis, Escherichia coli, Serratia marcensis e Candida albicans. Foi estabelecida a concentração mínima inibitória (CMI) através do método de difusão em meio sólido.Arrabidaea harleyi A.H. Gentry (Bignoniaceae) is an ornamental plant found in some regions of the Atlantic forest in Brazil. From its bark a mixture of verbascoside and isoverbascoside was isolated. This mixture was shown to be active againstStaphylococcus aureus, Micrococcus luteus, Bacillus subtilis, Bacillus mycoides, Enterecoccus faecalis, Escherichia coli, Serratia marcensisand Candida albicans. The minimal inhibitory concentration (MIC) was established by diffusion method

Cadernos Espinosanos (E-Journal)

Revista Brasileira de Ciências FarmacêuticasBrazilian Journal of Pharmaceutical Sciencesvol. 39, n. 1, jan./mar., 2003Antimicrobial activity of a mixture of two isomeric phenylpropanoidglycosides from Arrabidaea harleyi A.H. Gentry (Bignoniaceae)Cláudia Sampaio de Andrade Lima1, Elba Lúcia Cavalcanti de Amorim2, Kêsia Xisto da FonsecaRibeiro de Sena3, Alda de Andrade Chiappeta3, Xirley Pereira Nunes4, Maria de Fátima Agra4,Emidio Vasconcelos Leitão da-Cunha4,5, Marcelo Sobral da Silva4 and José Maria Barbosa-Filho4*1Departamento de Biofísica, Universidade Federal de Pernambuco, 2Departamento de Ciências Farmacêuticas,Universidade Federal de Pernambuco, 3Departamento de Antibióticos, Universidade Federal de Pernambuco,4Laboratório de Tecnologia Farmacêutica, Universidade Federal da Paraíba, 5Departamento de Farmácia eBiologia, Universidade Estadual da ParaíbaArrabidaea harleyi A.H. Gentry (Bignoniaceae) is an ornamentalplant found in some regions of the Atlantic forest in Brazil. Fromits bark a mixture of verbascoside and isoverbascoside was isolated.This mixture was shown to be active against Staphylococcus aureus,Micrococcus luteus, Bacillus subtilis, Bacillus mycoides,Enterecoccus faecalis, Escherichia coli, Serratia marcensis andCandida albicans. The minimal inhibitory concentration (MIC) wasestablished by diffusion method.Uniterms:• Arrabidaea harleyi• Bignoniaceae• Verbascoside• Isoverbascoside• Antimicrobial activity*Correspondence:J. M. Barbosa-FilhoLaboratório de TecnologiaFarmacêuticaUniversidade Federal da ParaíbaCaixa Postal 500958051-970 - João Pessoa - PBBrasilE-mail: jbarbosa@ltf.ufpb.brINTRODUCTIONThe family Bignoniaceae is essentially tropical, withmore than 100 genera and nearly 800 species encounteredmainly in the North part of South America. The plants aremostly trees and lianas, rarely as herbs (Brummitt, 1992).Only a few Brazilian species have been investigated so far.The genus Arrabidaea occurs in tropical America, fromMexico to Argentina. In Brazil most of species ofArrabidaea as A. triplinervia and A. pulchera are found inthe cerrado region. Other species e.g. A. agnus-castushave been found in the semi-desert regions ofNortheastern Brazil (Noronha, 1964), whereas the otherse.g. A. bilabiata and A. chica have been found in Amazo-nas, (Takemura et al., 1995; Zorn et al., 2001).Arrabidaea harleyi known as “cipó-do-mato” is aligneous vine, used in folk medicine as a fungicide,especially in the treatment of dandruff. It occurs along themargins of the Atlantic Forest, from the Brazilian state ofPiaui to Minas Gerais. This is the first report of thechemical and biological studies of this species and the firstreport of the title compounds in plants of this genus.MATERIAL AND METHODSPlant materialThe bark of Arrabidaea harleyi A.H. Gentry wascollected in March 2001 in a forest reserve in the city ofJoão Pessoa, state of Paraiba. A voucher specimen isdeposited in the Herbarium of the Laboratório deTecnologia Farmacêutica, Universidade Federal daParaiba, under the nº Agra 0364 (JPB).Extraction and IsolationAir dried ground bark of A. harleyi (3.5 Kg) wasexhaustively extracted with 5.0 L of 95% ethanol. TheC. S. A. Lima et al.78resulting extract (120.0 g) was dissolved in H2O:MeOH(7:3 v/v) and submitted to a partition between hexane (9.0g), CHCl3 (6.0 g), AcOEt (10.0 g) and n-BuOH (20.0 g).The AcOEt fraction was subjected to column chromato-graphy using silica gel (Merck) 70-230 mesh. Thesolvents were P.A. Merck: chloroform - methanol gradient(5 - 20%). 93 fractions of 100 mL were collected. Afteranalysis by TLC aluminium sheets silica gel-60 F254(Merck) developed with I2 or Neu/Peg reagent, somefractions were reunited yielding 19 fractions. The mainfraction (2.0 g) was an amorphous pale yellow powder. Its13C-APT NMR (CDCl3, 75 MHz) showed pairs of signalswith the same amplitude and these were denominated Aand B. (A): 131.1 (C1), 117.0 (C2), 144.0 (C3), 145.6(C4), 116.3 (C5), 121.0 (C6), 36.1 (C7), 71.5 (C8), 103.7(C1’), 76.0 (C2’), 81.0 (C3’), 70.1 (C4’), 75.6 (C5’), 62.1(C6’), 102.0 (C1”), 72.0 (C2”), 71.8 (C3”), 73.5 (C4”),70.0 (C5”), 18.4 (C6”), 127.3 (C1”’), 115.9 (C2’”), 146.1(C3”’), 149.0 (C4”’), 116.8 (C5”’), 123.0 (C6”’), 147.0(C7”’), 115.1 (C8”’), 167.6 (C=O). (B): 131.0 (C1), 117.0(C2), 144.0 (C3), 145.6 (C4), 116.3 (C5), 121.0 (C6), 36.0(C7), 71.4 (C8), 103.5 (C1’), 74.9 (C2’), 83.9 (C3’), 69.5(C4’), 74.7 (C5’), 64.2 (C6’), 101.9 (C1”), 72.0 (C2”),71.8 (C3”), 73.4 (C4”), 69.5 (C5”), 18.0 (C6”), 127.4(C1”’), 114.8 (C2”’), 146.1 (C3”’), 148.8 (C4”’), 115.9(C5”’), 122.7 (C6”’), 146.1 (C7”’), 115.0 (C8”’), 167.3(C=O).Antimicrobial qualitative assayA modified diffusion test (Bauer et al., 1966) wasused to determine the antimicrobial activity. The isomermixture was dissolved in DMSO and tested againstStaphylococcus aureus DAUFPE 01, Bacillus subtilisDAUFPE 16, Bacillus mycoides DAUFPE 14,Micrococcus luteus DAUFPE 06, Enterococcus faecalisDAUFPE 138 ,  Escherichia coli DAUFPE 224,Pseudomonas aeruginosa DAUFPE 39, Serratiamarcescens DAUFPE 398, Mycobacterium smegmatisDAUFPE 71, and Candida albicans DAUFPE 1007.These microorganisms were obtained from the CultureCollection of the Departamento de Antibióticos (Méllo,1988). From the fresh cultures of the microorganismsstandardized suspensions were prepared in physiologicalsolutions to comparison of 0.5 to MacFarland scale,equivalent at 107 UFC (Murray, 1995; Washington, 1972).Antimicrobial activities were evaluated by the diffusiontest on paper discs over Müller Hinton agar, glucose-yeastextract agar and sabouraud-dextrose media. Thesuspensions were spread on a surface of the medium inPetri dishes with Drigalsky’s loop (100 μL per dish). Paperdiscs at Whatman no 2 (6.0 mm diameter) were moistenedwith 20 μL of the mixture of isomers at the concentrationof 30 mg/mL, which is equivalent to 600 μg/disc. After aproper incubation at 30 or 35 ºC for 24 or 48 h, theinhibition zones around the discs were measured (Bauer etal., 1966). The tests were performed in triplicate and theresults were expressed in mm as the arithmetic media ofdiameters of the inhibition zones. The blank control wasperformed with DMSO. A standard solution of cephalexinand cetoconazol were used as a positive control to bacteriaand yeast, respectively.Minimal Inhibitory Concentration (MIC)A solution of 6 000 μg/mL of the mixture of A andB, was prepared and distributed in Petri dishes in differentvolumes (1.0, 0.5, 0.25, 0.125, 0.06, 0.03, 0.015 and0.007 mL), which contained 10 mL of the proper culturemedium. The microorganisms were spread and the disheswere incubated for 24 or 48 hours at 30 or 35 ºC, asshowed in Table 1 (Méllo et al., 1988).RESULTS AND DISCUSSIONPhytochemical analysis of EtOAc extract ofArrabidaea harleyi yielded a chromatographic fraction(F6), which when eluted in a TLC chromatogram andsprayed with Neu/Peg reagent or in the presence of I2 va-por showed only one spot. Analysis of the 300 MHz 1HNMR spectrum of this material, showed the signalscharacteristic of aromatic compounds linked to acarbohydrate group. The analysis of the 13C-APT NMRspectrum showed that all the signals were present in pairs,which suggested that the fraction was a mixture of twoisomers. A detailed spectral analysis and also a compa-rison with the literature data (Krebs, Habermehl, 1992),showed that these compounds were verbascoside(acteoside) (A), a glycosylated phenylpropanoid, andisoverbacoside (isoacteoside) (B). Microbiological assayswith this mixture showed that it possesses a wide spectrumof activity against Gram positive and Gram negativebacteria and also against yeast (Table I). The minimalinhibition concentrations (MIC) are also showed in TableI. Phytochemical studies of the family Bignoniaceae showthe presence of verbascoside and isoverbascoside in somegenera such as Deplanchea (Davioud et al., 1989),Jacaranda, Mussatia (Jimenez et al., 1987, 1988, 1989)Tecoma (Guiso, 1997) and Newboldia (Gafner, 1997), butthis is the first report of these compounds in the genusArrabidaea. Earlier reports describe the antineoplasicaction of verbascoside and isoverbascoside againstAntimicrobial activity of a mixture of two isomeric phenylpropanoid glycosides 79murine-388 (PS) lymphocytic leukemia (Pettit et al.,1990). It was determined the antiviral properties of thesecompounds obtained from Markhamia lutea (Kernan etal ., 1998). The pharmacological effects of thesecompounds obtained from Mussatia was described byCano et al.(1990).CONCLUSIONSOn a inseparable mixture of, two isomericcompounds was isolated. These compounds wereidentified as verbascoside (A) and isoverbascoside (B).Five Gram positive bacteria (Staphylococcus aureus,Micrococcus luteus, Bacillus subtilis, Bacillus mycoides,Enterococcus faecalis), two Gram negative bacteria(Escherichia coli and Serratia marcensis), and one yeast(Candida albicans) were shown to be sensitive to thismixture (A/B), whereas Pseudomonas aeruginosa (Gramnegative bacterium) and Mycobacterium smegmatis(alcohol-acid resistant bacterium) were found to beresistant. These results may support some traditional usesof this plant by local population.TABLE I - Inhibition zones and MIC of a mixture of verbascoside and isoverbascoside (A/B) against Gram positive, Gramnegative, alcohol-acid bacteria and yeastMicroorganisms  Inhibition zone (mm) 600 µg/disc  MIC (µg/mL)Bacteria A/B Cephalexin DMSO(30 µg)Staphylococcus aureus DAUFPE 01 (G +) 15 26 0 600Micrococcus luteus DAUFPE 06 (G +) 30 47 0 300Bacillus subtilis DAUFPE 16 (G +) 19 34 0 600Bacillus mycoides DAUFPE 14 (G +) 11 24 0 600Pseudomonas aeruginosa DAUFPE 39 (G -) 0 0 0 >600Mycobacterium smegmatis DAUFPE 71 (AAR) 0 0 0 >600Enterococcus faecalis DAUFPE 138 (G +) 15 11 0 600Escherichia coli DAUFPE 224 (G -) 15 18 0 600Serratia marcescens DAUFPE 398 (G -) 9 0 0 600Yeast Cetoconazol(300 µg)Candida albicans DAUFPE 1007 18 24 0 300(A/B) – not separated isomers mixture; (G +) - Gram positive bacterium; (G -) - Gram negative bacterium; (AAR) -Alcohol-acid resistant bacterium; (MIC) Minimal Inhibitory Concentration.C. S. A. Lima et al.80RESUMOAtividade antimicrobiana de uma mistura de doisisômeros de glicosídeos fenilpropanóides deArrabidaea harleyi A.H. Gentry (Bignoniaceae)Arrabidaea harleyi A.H. Gentry (Bignoniaceae) é umaplanta ornamental, encontrada em algumas regiões daMata Atlântica do Brasil. A partir das cascas do caule foiisolada a mistura dos isômeros verbascosídeo eisoverbascosídeo. A mistura mostrou-se ativa frente aStaphylococcus aureus, Micrococcus luteus, Bacillussubtilis, Bacillus mycoides, Enterococcus faecalis,Escherichia coli, Serratia marcensis e Candida albicans.Foi estabelecida a concentração mínima inibitória (CMI)através do método de difusão em meio sólido.UNITERMOS: Arrabidaea harleyi. Bignoniaceae.Verbascosídeo. Isoverbascosídeo. Atividade antimicro-biana.REFERENCESBAUER, A. W.; KIRBY, W. M. M.; SHERRIS, J. C.; TURC,M. Antibiotic susceptibility testing by a standardizedsingle disk method. Amer. J. Clin. Pathol., Philadelphia,v. 45, p. 493-496, 1966.BRUMMITT, R. K. Vascular plant families and genera.London: Royal Botanic Garden, 1992. p. 505–506.CANO, E.; VEIGA, M.; JIMENEZ, C.; RIGUERA, R.Pharmacological effects of three phenylpropanoidglycosides from Mussatia. Planta Med., Stuttgart, v. 56,n. 1, p. 24-26, 1990.DAVIOUD, E.; BAILLEUL, F.; DELAVEAU, P.; DEBRAY,M. M. Iridoid glycosides and phenylpropanoidglycosides from Deplanchea speciosa. Planta Med.,Stuttgart, v. 55, n. 1, p. 87, 1989.GAFNER, S.; WOLFENDER, J. L.; NIANGA, M.;HOSTETTMANN, K. Phenylpropanoid glycosidesfrom Newbouldia laevis roots. Phytochemistry, NewYork, v. 44, n. 4, p. 687-690, 1997.GUISO, M.; MARRA, C.; PICCIONI, F.; NICOLETTI, M.Iridoid and phenylpropanoid glycosides from Tecomacapensis. Phytochemistry, New York, v. 45, n. 1, p. 193-194, 1997.JIMENEZ, C.; VILLAVERDE, M. C.; RIGUERA, R.;CASTEDO, L.; STERMITZ, F. R. Three phenyl-propanoid glycosides from Mussatia. Phytochemistry,New York, v. 26, n. 6, p. 1805-1810, 1987.JIMENEZ, C.; VILLAVERDE, M. C.; RIGUERA, R.;CASTEDO, L.; STERMITZ, F. R. Five phenylpropanoidglycosides from Mussatia. Phytochemistry, New York, v.27, n. 9, p. 2947-2951, 1988.JIMENEZ, C.; VILLAVERDE, M. C.; RIGUERA, R.;CASTEDO, L.; STERMITZ, F. R. Phenylpropanoidglycosides from Mussatia hyacinthina. J. Nat. Prod.,Columbus, v. 52, n. 2, p. 408-410, 1989.KERNAN, M. R.; AMARQUAVE, A.; CHEN, J. L.; CHAN,J.; SESIN, D. F.; PARKINSON, N.; YE, Z. J.;BARRETT, M.; BALES, C.; STODDART, C. A.;SLOAN, B.; BLANC, P.; LIMBACH, C.; MRISHO, S.;ROZHON, E. J. Antiviral phenylpropanoid glycosidesfrom the medicinal plant Markhamia lutea. J. Nat. Prod.,Columbus, v. 61, n. 5, p. 564-570, 1998.KREBS, H. C.; HABERMEHL, G. G. Two dimensionalNMR spectroscopy of glycosides. Magn. Reson. Chem.,Chichestor, v. 30, p. 556-559, 1992.MÉLLO, B. R. Catálogo da coleção de microrganismos.2.ed. Recife: UFPE/Departamento de Antibióticos, 1988.p. 45.MURRAY, P. R.; BARON, E. J.; PFALLER, M. A.;TENOVER, F. C.; YOLKEN, R. H. Manual of clinicalmicrobiology. 6. ed. Washington: American Society forMicrobiology, 1995. p.165.NORONHA, H. A propósito dos cipós medicinais do Brasil.Rev. Bras. Farm., Rio de Janeiro, v. 45, n. 3, p. 151-159,1964.PETTIT, G. R.; NUMATA, A.; TAKEMURA, T.; ODE, R.H.; NARULA, A. S.; CRAGG, G. M.; PASE, C. P.Antineoplasic agents 107. Isolation of acteoside andisoacteoside from Castilleja linariaefolia. J. Nat. Prod.,Columbus, v. 53, n. 2, p. 456-458, 1990.TAKEMURA, O. S.; IINUMA, M.; TOSA, H.; MIGUEL, O.G.; MOREIRA, E. A.; NOZAWA, Y. A. Flavone fromleaves of Arrabidaea chica f. cuprea. Phytochemistry,New York, v. 38, n. 5, p. 1299-1300, 1995.Antimicrobial activity of a mixture of two isomeric phenylpropanoid glycosides 81WASHINGTON, J. A.; WARREN, E.; KARLSON, A. G.Stability of barium sulfate turbidity standards. Appl.Microbiol., Washington, v. 24, p. 1013, 1972.ZORN, B.; GARCÍA-PIÑERES, A. J.; CASTRO, V.;MURILLO, R.; MORA, G.; MERFORT, I. 3-Desoxyanthocyanidins from Arrabidaea chica.Phytochemistry, New York, v. 56, n. 8, p. 831-835, 2001.Recebido para publicação em 04 de março de 2002.

Atividade antimicrobiana de uma mistura de dois isômeros de glicosídeos fenilpropanóides de Arrabidaea harleyi A.H. Gentry (Bignoniaceae)

https://www.revistas.usp.br/rbcf/article/download/43842/47463

Atividade antimicrobiana de uma mistura de dois isômeros de glicosídeos fenilpropanóides de Arrabidaea harleyi A.H. Gentry (Bignoniaceae)

Abstract

Similar works

Full text

Available Versions

Cadernos Espinosanos (E-Journal)