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Coumarins, coumarinolignoids and terpenes from Protium heptaphyllum
Biochemical Systematics and Ecology 30 (2002) 685–687 www.elsevier.com/locate/biochemsyseco
Coumarins, coumarinolignoids and terpenes from Protium heptaphyllum Elisangela Xavier Almeida a, Lucia M. Conserva a,*, Rosangela P. Lyra Lemos b a
Departamento de Quı´mica, Universidade Federal de Alagoas, 57072-970, Maceio´, AL, Brazil b Instituto do Meio Ambiente do Estado de Alagoas, 57017-320, Maceio´, AL, Brazil Received 26 March 2001; accepted 7 July 2001
Keywords: Protium heptaphyllum; Burseraceae; Coumarins; Coumarinolignoids; Terpenes; Phenylpropanoid derivatives
1. Subject and source Protium heptaphyllum March (Burseraceae), known as “breu branco, alme´cega do Brasil, alme´sca, goma-lima˜o, etc.”, is distributed in the north and northeast regions of Brazil (Lorenzi, 1998). Stems, including barks, and leaves were collected in Marechal Deodoro, Alagoas, Brazil, in October 1994 and identified by Rosangela Pereira de Lyra Lemos of the Departamento de Botaˆnica do Instituto do Meio Ambiente do Estado de Alagoas (IMA/AL), where a voucher specimen was deposited (MAC8691).
2. Previous work Previous studies on the secondary metabolites of Protium heptaphyllum and related species have shown monoterpenes (Gottlieb et al., 1981; Siani et al., 1999), triterpenes (Maia et al., 2000), lignans (Siani et al., 1998; Siqueira et al., 1995) and simple phenylpropanoid derivatives (Pernet, 1972) being the most abundant constituents. A *
Corresponding author. Tel.: +55-82-214-1384; fax: +55-82-214-1615. E-mail address: [email protected] (L.M. Conserva).
0305-1978/02/$ - see front matter 2002 Elsevier Science Ltd. All rights reserved. PII: S 0 3 0 5 - 1 9 7 8 ( 0 1 ) 0 0 1 3 0 - 2
686
E.X. Almeida et al. / Biochemical Systematics and Ecology 30 (2002) 685–687
coumarinolignoid has been isolated from Protium opacum Swart (Zoghbi et al., 1981; Marques and Yoshida, 1990). 3. Present study The air-dried and powdered stem and bark (3290 g) and leaves (380 g) were extracted with 90% EtOH at room temperature. The crude extracts (stems and barks: 336 g, 10.2%; leaves: 43.9 g, 11.6%) were resuspended in MeOH/H2O (3:2) solution and extracted successively with n-C6H14, CHCl3 and EtOAc. The CHCl3 residue (11.5 g, 0.35%) from the stem and barks was chromatographed on a silica gel column. Elution with n-C6H14/CH2Cl2 containing increasing amounts of CH2Cl2 afforded after gel permeation (Sephadex LH-20 from MeOH) and TLC prep. [C6H6/AcOEt (8.5:1.5)] p-coumaric ethyl ester (34 mg, 0.001%), scopoletin (19 mg, 0.0005%) and fraxetin (11 mg, 0.0003%). The remaining fractions after centrifugation (1 h at 7000 rpm from MeOH) followed by repeated recrystallizations from MeOH afforded propacin (23 mg, 0.0007%), cleomiscosin A (35 mg, 0.0012%) and 3-O-β-D-glycopyranosyl-β-sitosterol (194 mg, 0.006%). The CHCl3 residue (5.6 g, 1.47%) from the leaves was suspended in aq. NaOH 4% and extracted with n-C6H14 to yield a neutral fraction (2.5 g, 0.66%). This was further chromatographed on a silica gel column. Elution with n-C6H14/EtOAc containing increasing amounts of EtOAc following recrystallizations from n-C6H14 afforded lupeol (148 mg, 0.039%) and a mixture containing sitosterol and stigmasterol (36 mg, 0.0095%). The spectral (IR, MS, 1H and 13C NMR) data of lupeol (Mahato and Kundu, 1994), coumarins (Marques and Yoshida, 1990), coumarinolignoids (Zoghbi et al., 1981; Arnoldi et al., 1984; Marques and Yoshida, 1990; Chatterjee et al., 1994) were found to be similar to those recorded earlier. 2D NMR experiments (COSY, HMQC and HMBC) were used to distinguish the possibility of the regioisomers. 4. Chemotaxonomic significance According to Dahlgren’s system, the order Rutales contains the families Rutaceae, Cneoraceae, Surianaceae, Simaroubaceae, Meliaceae and Burseraceae (Dahlgren, 1980). In this order, the majority of reports of coumarins have been from the Rutaceae. To our knowledge, no coumarin has been isolated so far from Burseraceae species. On the other hand, the occurrence of coumarinolignoid is not uncommon in the Burseraceae species. Propacin was previously reported in Protium opacum Swart (Zoghbi et al., 1981; Marques and Yoshida, 1990), but this is the first time that cleomiscosin A has been found in Burseraceae. Acknowledgements This work was supported by Conselho Nacional de Desenvolvimento Cientı´fico e Tecnolo´ gico (CNPq/PADCT II) and CAPES. The authors are grateful to Dr Mar-
E.X. Almeida et al. / Biochemical Systematics and Ecology 30 (2002) 685–687
687
celo Sobral da Silva and Dr Emı´dio de V. Cunha Leita˜ o, Laborato´ rio de Tecnologia Farmaceˆ utica da Universidade Federal da Paraı´ba, Dr Edilberto Rocha Silveira, Centro Nordestino de Aplicac¸ a˜ o e Uso de RMN da Universidade Federal do Ceara´ (CENAUREN) for the NMR spectra and MSc. Eloisa Helena A. Andrade for MS spectra.
References Arnoldi, A., Arnone, A., Merlini, L., 1984. Heterocycles 22, 1537. Chatterjee, A., Das, P.C., Joshi, P.C., Mandal, S., 1994. J. Indian Chem. Soc. 71, 475. Dahlgren, R.M.T., 1980. Bot. J. Linn. Soc. 80, 91. Gottlieb, O.R., Koketsu, M., Magalha˜ es, M.T., Maia, J.G.S., Mendes, P.H., Rocha, A.I., Silva, M.L., wilberg, V.C., 1981. Acta Amazoˆ nica 11, 143. ´ rvores Brasileiras: manual de identificac¸ a˜ o e cultivo de plantas arbo´ reas nativas Lorenzi, H. (Ed.), 1998. A do Brasil, vol. 1. Instituto Plantarum de Estudos da Flora, Nova Odessa, 76. Mahato, S.B., Kundu, A.P., 1994. Phytochemistry 37, 1517. Maia, R.M., Barbosa, P.R., Cruz, F.G., Roque, N.F., Fascio, M., 2000. Quı´mica Nova 25, 623. Marques, M.O.M., Yoshida, M., 1990. Quı´mica Nova 13, 245. Pernet, R., 1972. Lloydia 35, 281. Siani, A.C., Ramos, M.F.S., Menezes-de-Lima, O., Ribeiro-dos-Santos, R., Fernandez-Ferreira, E., Soares, R.O.A., Rosas, E.C., Susunaga, G.S., Guimara˜ es, A.C., Zoghbi, M.G.B., Henriques, M.G.M.O., 1999. J. Ethnopharm. 66, 57. Siani, A.C., Zoghbi, M.G.B., Wolter, E.L.A., Vencato, I., 1998. J. Nat. Prod. 61, 796. Siqueira, J.B.G., Zoghbi, M.G.B., Cabral, J.A., Filho, W.W., 1995. J. Nat. Prod. 58, 730. Zoghbi, M.G.B., Roque, N.F., Gottlieb, O.R., 1981. Phytochemistry 20, 180.
Coumarins, coumarinolignoids and terpenes from Protium heptaphyllum Elisangela Xavier Almeida a, Lucia M. Conserva a,*, Rosangela P. Lyra Lemos b a
Departamento de Quı´mica, Universidade Federal de Alagoas, 57072-970, Maceio´, AL, Brazil b Instituto do Meio Ambiente do Estado de Alagoas, 57017-320, Maceio´, AL, Brazil Received 26 March 2001; accepted 7 July 2001
Keywords: Protium heptaphyllum; Burseraceae; Coumarins; Coumarinolignoids; Terpenes; Phenylpropanoid derivatives
1. Subject and source Protium heptaphyllum March (Burseraceae), known as “breu branco, alme´cega do Brasil, alme´sca, goma-lima˜o, etc.”, is distributed in the north and northeast regions of Brazil (Lorenzi, 1998). Stems, including barks, and leaves were collected in Marechal Deodoro, Alagoas, Brazil, in October 1994 and identified by Rosangela Pereira de Lyra Lemos of the Departamento de Botaˆnica do Instituto do Meio Ambiente do Estado de Alagoas (IMA/AL), where a voucher specimen was deposited (MAC8691).
2. Previous work Previous studies on the secondary metabolites of Protium heptaphyllum and related species have shown monoterpenes (Gottlieb et al., 1981; Siani et al., 1999), triterpenes (Maia et al., 2000), lignans (Siani et al., 1998; Siqueira et al., 1995) and simple phenylpropanoid derivatives (Pernet, 1972) being the most abundant constituents. A *
Corresponding author. Tel.: +55-82-214-1384; fax: +55-82-214-1615. E-mail address: [email protected] (L.M. Conserva).
0305-1978/02/$ - see front matter 2002 Elsevier Science Ltd. All rights reserved. PII: S 0 3 0 5 - 1 9 7 8 ( 0 1 ) 0 0 1 3 0 - 2
686
E.X. Almeida et al. / Biochemical Systematics and Ecology 30 (2002) 685–687
coumarinolignoid has been isolated from Protium opacum Swart (Zoghbi et al., 1981; Marques and Yoshida, 1990). 3. Present study The air-dried and powdered stem and bark (3290 g) and leaves (380 g) were extracted with 90% EtOH at room temperature. The crude extracts (stems and barks: 336 g, 10.2%; leaves: 43.9 g, 11.6%) were resuspended in MeOH/H2O (3:2) solution and extracted successively with n-C6H14, CHCl3 and EtOAc. The CHCl3 residue (11.5 g, 0.35%) from the stem and barks was chromatographed on a silica gel column. Elution with n-C6H14/CH2Cl2 containing increasing amounts of CH2Cl2 afforded after gel permeation (Sephadex LH-20 from MeOH) and TLC prep. [C6H6/AcOEt (8.5:1.5)] p-coumaric ethyl ester (34 mg, 0.001%), scopoletin (19 mg, 0.0005%) and fraxetin (11 mg, 0.0003%). The remaining fractions after centrifugation (1 h at 7000 rpm from MeOH) followed by repeated recrystallizations from MeOH afforded propacin (23 mg, 0.0007%), cleomiscosin A (35 mg, 0.0012%) and 3-O-β-D-glycopyranosyl-β-sitosterol (194 mg, 0.006%). The CHCl3 residue (5.6 g, 1.47%) from the leaves was suspended in aq. NaOH 4% and extracted with n-C6H14 to yield a neutral fraction (2.5 g, 0.66%). This was further chromatographed on a silica gel column. Elution with n-C6H14/EtOAc containing increasing amounts of EtOAc following recrystallizations from n-C6H14 afforded lupeol (148 mg, 0.039%) and a mixture containing sitosterol and stigmasterol (36 mg, 0.0095%). The spectral (IR, MS, 1H and 13C NMR) data of lupeol (Mahato and Kundu, 1994), coumarins (Marques and Yoshida, 1990), coumarinolignoids (Zoghbi et al., 1981; Arnoldi et al., 1984; Marques and Yoshida, 1990; Chatterjee et al., 1994) were found to be similar to those recorded earlier. 2D NMR experiments (COSY, HMQC and HMBC) were used to distinguish the possibility of the regioisomers. 4. Chemotaxonomic significance According to Dahlgren’s system, the order Rutales contains the families Rutaceae, Cneoraceae, Surianaceae, Simaroubaceae, Meliaceae and Burseraceae (Dahlgren, 1980). In this order, the majority of reports of coumarins have been from the Rutaceae. To our knowledge, no coumarin has been isolated so far from Burseraceae species. On the other hand, the occurrence of coumarinolignoid is not uncommon in the Burseraceae species. Propacin was previously reported in Protium opacum Swart (Zoghbi et al., 1981; Marques and Yoshida, 1990), but this is the first time that cleomiscosin A has been found in Burseraceae. Acknowledgements This work was supported by Conselho Nacional de Desenvolvimento Cientı´fico e Tecnolo´ gico (CNPq/PADCT II) and CAPES. The authors are grateful to Dr Mar-
E.X. Almeida et al. / Biochemical Systematics and Ecology 30 (2002) 685–687
687
celo Sobral da Silva and Dr Emı´dio de V. Cunha Leita˜ o, Laborato´ rio de Tecnologia Farmaceˆ utica da Universidade Federal da Paraı´ba, Dr Edilberto Rocha Silveira, Centro Nordestino de Aplicac¸ a˜ o e Uso de RMN da Universidade Federal do Ceara´ (CENAUREN) for the NMR spectra and MSc. Eloisa Helena A. Andrade for MS spectra.
References Arnoldi, A., Arnone, A., Merlini, L., 1984. Heterocycles 22, 1537. Chatterjee, A., Das, P.C., Joshi, P.C., Mandal, S., 1994. J. Indian Chem. Soc. 71, 475. Dahlgren, R.M.T., 1980. Bot. J. Linn. Soc. 80, 91. Gottlieb, O.R., Koketsu, M., Magalha˜ es, M.T., Maia, J.G.S., Mendes, P.H., Rocha, A.I., Silva, M.L., wilberg, V.C., 1981. Acta Amazoˆ nica 11, 143. ´ rvores Brasileiras: manual de identificac¸ a˜ o e cultivo de plantas arbo´ reas nativas Lorenzi, H. (Ed.), 1998. A do Brasil, vol. 1. Instituto Plantarum de Estudos da Flora, Nova Odessa, 76. Mahato, S.B., Kundu, A.P., 1994. Phytochemistry 37, 1517. Maia, R.M., Barbosa, P.R., Cruz, F.G., Roque, N.F., Fascio, M., 2000. Quı´mica Nova 25, 623. Marques, M.O.M., Yoshida, M., 1990. Quı´mica Nova 13, 245. Pernet, R., 1972. Lloydia 35, 281. Siani, A.C., Ramos, M.F.S., Menezes-de-Lima, O., Ribeiro-dos-Santos, R., Fernandez-Ferreira, E., Soares, R.O.A., Rosas, E.C., Susunaga, G.S., Guimara˜ es, A.C., Zoghbi, M.G.B., Henriques, M.G.M.O., 1999. J. Ethnopharm. 66, 57. Siani, A.C., Zoghbi, M.G.B., Wolter, E.L.A., Vencato, I., 1998. J. Nat. Prod. 61, 796. Siqueira, J.B.G., Zoghbi, M.G.B., Cabral, J.A., Filho, W.W., 1995. J. Nat. Prod. 58, 730. Zoghbi, M.G.B., Roque, N.F., Gottlieb, O.R., 1981. Phytochemistry 20, 180.