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Triterpenoid saponins and iridoid glycosides from the aerial parts of Cephalaria pastricensis
Biochemical Systematics and Ecology 34 (2006) 890e893 www.elsevier.com/locate/biochemsyseco
Triterpenoid saponins and iridoid glycosides from the aerial parts of Cephalaria pastricensis Dejan GoCevac a, Boris Mandic´ b, Vlatka Vajs a, Vele Tesˇevic´ b, Nebojsˇa Menkovic´ c, PeCa Janac´kovic´ d, Slobodan Milosavljevic´ b,* a
Institute for Chemistry, Technology and Metallurgy, Njegosˇeva 12, Belgrade, Serbia and Montenegro b Faculty of Chemistry, Studentski trg 16, Belgrade, Serbia and Montenegro c Institute for Medicinal Plant Research ‘‘Dr. Josif Pancic´’’, Tadeusˇa Kosˇc´usˇka 1, Belgrade, Serbia and Montenegro d Faculty of Biology, Studentski trg 16, Belgrade, Serbia and Montenegro Received 22 June 2005; accepted 22 May 2006
Keywords: Cephalaria pastricensis; Dipsacaceae; Triterpenoid saponins; Iridoid glycosides
1. Subject and source The genus Cephalaria Schrad. (Dipsacaceae family) comprises about 30 species, distributed mainly in the Mediterranean region, Balkan peninsula and the Middle East. Cephalaria pastricensis Do¨rfler & Hayek is an endemic species widespread on mountain parts of North Albania, Bosnia and Herzegovina and Serbia and Montenegro (Diklic´, 1973). The plant material was collected in July 2000 at Veliki Stolac, mountain Zvijezda, SerbianeBosnian border. A voucher specimen (CP28072003) was deposited in the Herbarium of the Faculty of Biology, University of Belgrade.
2. Previous work Two luteolin type flavonoid glycosides with antiradical activity were isolated from flowers of C. pastricensis (GoCevac et al., 2004).
* Corresponding author. Tel.: þ381 11 630 474; fax: þ381 11 636 061. E-mail address: [email protected] (S. Milosavljevic´). 0305-1978/$ - see front matter Ó 2006 Elsevier Ltd. All rights reserved. doi:10.1016/j.bse.2006.05.015
D. GoC evac et al. / Biochemical Systematics and Ecology 34 (2006) 890e893
891
Investigation of an additional seven species of the genus revealed triterpenoid saponins of hederagenin or oleanolic acid type (Movsumov et al., 1975; Zviadadze et al., 1976, 1980; Alankus-Caliskan and Anil, 1994, 1995; Kirmizigul and Anil, 1994, 2002; Kirmizigul et al., 1996; Kirmizigul and Rose, 1997), quercetin and luteolin type flavonoid glycosides (Zemtsova and Bandyukova, 1970; Bouillant et al., 1972; Movsumov and Aliev, 1975), as well as alkaloids (Aliev and Movsumov, 1975). From Cephalaria ambrosioides, Pasi et al. (2002) isolated iridoid glycosides, hydroxycinnamic esters and an unusual lignan with two tertiary hydroxyl groups.
3. Present study The air-dried leaves of C. pastricensis (450 g) were extracted with 90% MeOH, (3 600 ml) at room temperature. The crude, slurry extract was partitioned between H2O and CH3Cl. Water layer was then extracted with n-BuOH, giving 35 g of residue, subsequently subjected to Sephadex LH-20 CC eluting with MeOH. A saponin rich fraction was submitted to silica gel CC, and eluted with CH3Cl/MeOH/H2O (13/7/1) to yield 118 fractions (frs1e118). Further purification of frs15e25 on silica gel CC using CH3Cl/MeOH/H2O (20/10/1) afforded 150 mg of 3-O-[b-D-xylop yranosyl(1 / 3)-a-L-rhamnopyranosyl(1 / 2)-a-L-arabinopyranosyl]-28-O-[b-D-glucopyranosyl]-hederagenin (1), named decaisoside D (Kong et al., 1993). From frs40e52 70 mg of 3-O-[a-L-rhamnopyranosyl(1 / 2)-a-L-arabinopyranosyl]-28-O-[b-D-glucopyranosyl(1 / 6)-b-D-glucopyranosyl]-hederagenin (2), named dipsacoside B (Mukhamedziev et al., 1971; Kong et al., 1993) was isolated. Purification of frs61e99 using Low-pressure LC (LiChroprepÒ Si 60 (40e63 mm) using CH3Cl/MeOH/H2O (20/10/1.9) yielded 50 mg of 3-O-[b-D-xylopyranosyl(1 / 3)-a-L-rhamnopyranosyl(1 / 2)-a-L-arabinopyranosyl]-28-O-[b-D-glucopyranosyl(1 / 6)-b-D-glucopyranosyl]-hederagenin (3), named decaisoside E (Kong et al., 1993), and 30 mg of 3-O-[b-D-glucopyranosyl(1 / 3)-a-L-rhamnopyranosyl(1 / 2)-a-L-arabinopyranosyl]-28-O-[b-D-glucopyranosyl(1 / 6)-b-D-glucopyranosyl]-hederagenin, 4 (Penders and Delaude, 1994). For structural elucidations of these compounds, both chemical and instrumental methods were used. Identification of sugars was carried out after acid and base hydrolysis. Configurations, positions of sugar connection and aglycon structure (hederagenin) were determined by spectroscopic methods (IR, ESI or MALDI MS, 1H and 13C NMR, 2D NMR), and confirmed by comparison with the data in the literature. The air-dried flowers of C. pastricensis (150 g) were extracted with 90% MeOH (3 600 ml). Residue was subjected to Sephadex LH-20 CC, and eluted with MeOH. Fractions rich in iridoid glycosides were further purified on silica gel CC, then eluted with CH3Cl/MeOH/H2O (40/13/1), and with the same solvents using increasing polarity ratios of 40/20/2 and 40/22/4. Combined fractions eluted with 40/13/1 ratio were rechromatographed on a Low-pressure LC (LiChroprepÒ Si 60, 40e63 mm), using CH3Cl/MeOH/H2O (40/10/1) solvent system, to yield 15 mg of secoiridoid glycoside sweroside (5) and 80 mg of iridoid glycoside loganin (6). From the fractions eluted with 40/22/4 ratio of solvents, 14 mg of loganic acid (7) was isolated. Structures of these iridoids were established according to 1H and 13C NMR data, and confirmed by comparison with those in the literature (Chapelle, 1976; Bianco et al., 1983). The hemolytic activity of saponins 1e4 was tested on bovine erythrocytes (Lemmich et al., 1995), as well as their cytotoxic activity using brine shrimps (Artemia salina) bioassay (Meyer et al., 1982). Saponins 1e4 showed 58%, 42%, 60% and 84% of hemolysis at concentration of 1 mg/ml, respectively. None of them showed cytotoxic activity on brine shrimps at the same concentrations.
4. Chemotaxonomic significance The genus Cephalaria is a member of Dipsacaceae, a taxonomically complex family (Verlaque, 1984). All the isolated iridoides were found for the first time in C. pastricensis. Sweroside, loganin and loganic acid characterize the family Dipsacaceae and could be of value as chemotaxonomic markers of the family (Jensen et al., 1979; Tomita and Mouri, 1996; Perdetzoglou et al., 2000; Horn et al., 2001; Tomassini et al., 2004). Saponins 1e4 were found for the first time in C. pastricensis. With the exception of dipsacoside B (2) this is the first report of their presence in Dipsacaceae family.
D. GoC evac et al. / Biochemical Systematics and Ecology 34 (2006) 890e893
892
O
OR2
R1O CH2OH
1 2 3 4
R1=
-D-Xyl(1-3)- -L-Rha(1-2)- -L-Ara-
R2=
-D-Glc-
R1=
-L-Rha(1-2)- -L-Ara-
R2=
-D-Glc(1-6)- -D-Glc-
R1=
-D-Xyl(1-3)- -L-Rha(1-2)- -L-Ara-
R2=
-D-Glc(1-6)- -D-Glc-
R1=
-D-Glc(1-3)- -L-Rha(1-2)- -L-Ara-
R2=
-D-Glc(1-6)- -D-Glc-
O
O
O
H
O
O
H
H
HO
HO O
O
H
O
H OGlc
OGlc
5
OH
6
OGlc
7
References Alankus-Caliskan, O., Anil, H., 1994. J. Nat. Prod. 57, 1001. Alankus-Caliskan, O., Anil, H., 1995. Phytochemistry 38, 1493. Aliev, A.M., Movsumov, I.S., 1975. Khim. Prir. Soedin. 11, 667. Bianco, A., Passacantilli, P., Polidori, G., Nicoletti, M., Messana, I., 1983. Gazz. Chim. Ital. 113, 829. Bouillant, M.L., Chopin, J., Plouvier, V., 1972. Phytochemistry 11, 1858. Chapelle, J.P., 1976. Planta Med. 29, 268. Diklic´, N., 1973. Dipsacaceae. In: Josifovic´, M. (Ed.), Flore de la Republique Socialiste de Serbie V. Academie Serbe des Sciences et des Arts, Belgrade, pp. 547e548. GoCevac, D., Vajs, V., Menkovic´, N., Tesˇevic´, V., Janac´kovic´, P., Milosavljevic´, S., 2004. J. Serb. Chem. Soc. 69, 883. Horn, M.M., Drewes, S.E., Brown, N.J., Munro, O.Q., Meyer, J.J.M., Mathekga, A.D.M., 2001. Phytochemistry 57, 51. Jensen, S.R., Lyse-Petersen, S.E., Nielsen, B.J., 1979. Phytochemistry 18, 273. Kirmizigul, S., Anil, H., 1994. Phytochemistry 36, 1555. Kirmizigul, S., Anil, H., 2002. Turk. J. Chem. 26, 947. Kirmizigul, S., Anil, H., Rose, M., 1996. Phytochemistry 39, 1171. Kirmizigul, S., Rose, M., 1997. Planta Med. 63, 51. Kong, J., Li, X., Wei, B., Yang, C., 1993. Phytochemistry 33, 427.
D. GoC evac et al. / Biochemical Systematics and Ecology 34 (2006) 890e893
893
Lemmich, E., Cornett, C., Jørstian, C., Knudsen, A.D., Thiilborg, A., Furu, P., Olsen, C., Salih, A., 1995. Phytochemistry 39, 63. Meyer, B.N., Ferrigni, N.R., Putnam, J.E., Jacobsen, L.B., Nichols, D.E., McLaughlin, J.L., 1982. Planta Med. 45, 31. Movsumov, I.S., Aliev, A.M., 1975. Khim. Prir. Soedin. 11, 804. Movsumov, I.S., Aliev, A.M., Kondratenko, E.S., Abubakirov, N.K., 1975. Khim. Prir. Soedin. 11, 519. Mukhamedziev, M.M., Alimbaeva, P.K., Gorovits, T.T., Abubakirov, N.K., 1971. Khim. Prir. Soedin. 7, 153. Pasi, S., Aligiannis, N., Skaltsounis, A., Chinou, I., 2002. Nat. Prod. Lett. 16, 365. Penders, A., Delaude, C., 1994. Carbohydr. Res. 263, 79. Perdetzoglou, D.K., Efthimiopoulos, C., Harvala, C., 2000. Plant Biosys. 134, 67. Tomassini, L., Foddai, S., Nicoletti, M., 2004. Biochem. Syst. Ecol. 32, 1083. Tomita, H., Mouri, Y., 1996. Phytochemistry 42, 239. Verlaque, R., 1984. A biosystematic and phylogenetic study of the Dipsacaceae. In: Grant, W. (Ed.), Plant Biosystematics. Academic Press, Toronto, pp. 307e320. Zemtsova, G.N., Bandyukova, V.A., 1970. Khim. Prir. Soedin. 6, 630. Zviadadze, L.D., Dekanosidze, G.E., Kemertelidze, E.P., 1976. Khim. Prir. Soedin. 13, 423. Zviadadze, L.D., Dekanosidze, G.E., Kemertelidze, E.P., 1980. Khim. Prir. Soedin. 3, 423.
Triterpenoid saponins and iridoid glycosides from the aerial parts of Cephalaria pastricensis Dejan GoCevac a, Boris Mandic´ b, Vlatka Vajs a, Vele Tesˇevic´ b, Nebojsˇa Menkovic´ c, PeCa Janac´kovic´ d, Slobodan Milosavljevic´ b,* a
Institute for Chemistry, Technology and Metallurgy, Njegosˇeva 12, Belgrade, Serbia and Montenegro b Faculty of Chemistry, Studentski trg 16, Belgrade, Serbia and Montenegro c Institute for Medicinal Plant Research ‘‘Dr. Josif Pancic´’’, Tadeusˇa Kosˇc´usˇka 1, Belgrade, Serbia and Montenegro d Faculty of Biology, Studentski trg 16, Belgrade, Serbia and Montenegro Received 22 June 2005; accepted 22 May 2006
Keywords: Cephalaria pastricensis; Dipsacaceae; Triterpenoid saponins; Iridoid glycosides
1. Subject and source The genus Cephalaria Schrad. (Dipsacaceae family) comprises about 30 species, distributed mainly in the Mediterranean region, Balkan peninsula and the Middle East. Cephalaria pastricensis Do¨rfler & Hayek is an endemic species widespread on mountain parts of North Albania, Bosnia and Herzegovina and Serbia and Montenegro (Diklic´, 1973). The plant material was collected in July 2000 at Veliki Stolac, mountain Zvijezda, SerbianeBosnian border. A voucher specimen (CP28072003) was deposited in the Herbarium of the Faculty of Biology, University of Belgrade.
2. Previous work Two luteolin type flavonoid glycosides with antiradical activity were isolated from flowers of C. pastricensis (GoCevac et al., 2004).
* Corresponding author. Tel.: þ381 11 630 474; fax: þ381 11 636 061. E-mail address: [email protected] (S. Milosavljevic´). 0305-1978/$ - see front matter Ó 2006 Elsevier Ltd. All rights reserved. doi:10.1016/j.bse.2006.05.015
D. GoC evac et al. / Biochemical Systematics and Ecology 34 (2006) 890e893
891
Investigation of an additional seven species of the genus revealed triterpenoid saponins of hederagenin or oleanolic acid type (Movsumov et al., 1975; Zviadadze et al., 1976, 1980; Alankus-Caliskan and Anil, 1994, 1995; Kirmizigul and Anil, 1994, 2002; Kirmizigul et al., 1996; Kirmizigul and Rose, 1997), quercetin and luteolin type flavonoid glycosides (Zemtsova and Bandyukova, 1970; Bouillant et al., 1972; Movsumov and Aliev, 1975), as well as alkaloids (Aliev and Movsumov, 1975). From Cephalaria ambrosioides, Pasi et al. (2002) isolated iridoid glycosides, hydroxycinnamic esters and an unusual lignan with two tertiary hydroxyl groups.
3. Present study The air-dried leaves of C. pastricensis (450 g) were extracted with 90% MeOH, (3 600 ml) at room temperature. The crude, slurry extract was partitioned between H2O and CH3Cl. Water layer was then extracted with n-BuOH, giving 35 g of residue, subsequently subjected to Sephadex LH-20 CC eluting with MeOH. A saponin rich fraction was submitted to silica gel CC, and eluted with CH3Cl/MeOH/H2O (13/7/1) to yield 118 fractions (frs1e118). Further purification of frs15e25 on silica gel CC using CH3Cl/MeOH/H2O (20/10/1) afforded 150 mg of 3-O-[b-D-xylop yranosyl(1 / 3)-a-L-rhamnopyranosyl(1 / 2)-a-L-arabinopyranosyl]-28-O-[b-D-glucopyranosyl]-hederagenin (1), named decaisoside D (Kong et al., 1993). From frs40e52 70 mg of 3-O-[a-L-rhamnopyranosyl(1 / 2)-a-L-arabinopyranosyl]-28-O-[b-D-glucopyranosyl(1 / 6)-b-D-glucopyranosyl]-hederagenin (2), named dipsacoside B (Mukhamedziev et al., 1971; Kong et al., 1993) was isolated. Purification of frs61e99 using Low-pressure LC (LiChroprepÒ Si 60 (40e63 mm) using CH3Cl/MeOH/H2O (20/10/1.9) yielded 50 mg of 3-O-[b-D-xylopyranosyl(1 / 3)-a-L-rhamnopyranosyl(1 / 2)-a-L-arabinopyranosyl]-28-O-[b-D-glucopyranosyl(1 / 6)-b-D-glucopyranosyl]-hederagenin (3), named decaisoside E (Kong et al., 1993), and 30 mg of 3-O-[b-D-glucopyranosyl(1 / 3)-a-L-rhamnopyranosyl(1 / 2)-a-L-arabinopyranosyl]-28-O-[b-D-glucopyranosyl(1 / 6)-b-D-glucopyranosyl]-hederagenin, 4 (Penders and Delaude, 1994). For structural elucidations of these compounds, both chemical and instrumental methods were used. Identification of sugars was carried out after acid and base hydrolysis. Configurations, positions of sugar connection and aglycon structure (hederagenin) were determined by spectroscopic methods (IR, ESI or MALDI MS, 1H and 13C NMR, 2D NMR), and confirmed by comparison with the data in the literature. The air-dried flowers of C. pastricensis (150 g) were extracted with 90% MeOH (3 600 ml). Residue was subjected to Sephadex LH-20 CC, and eluted with MeOH. Fractions rich in iridoid glycosides were further purified on silica gel CC, then eluted with CH3Cl/MeOH/H2O (40/13/1), and with the same solvents using increasing polarity ratios of 40/20/2 and 40/22/4. Combined fractions eluted with 40/13/1 ratio were rechromatographed on a Low-pressure LC (LiChroprepÒ Si 60, 40e63 mm), using CH3Cl/MeOH/H2O (40/10/1) solvent system, to yield 15 mg of secoiridoid glycoside sweroside (5) and 80 mg of iridoid glycoside loganin (6). From the fractions eluted with 40/22/4 ratio of solvents, 14 mg of loganic acid (7) was isolated. Structures of these iridoids were established according to 1H and 13C NMR data, and confirmed by comparison with those in the literature (Chapelle, 1976; Bianco et al., 1983). The hemolytic activity of saponins 1e4 was tested on bovine erythrocytes (Lemmich et al., 1995), as well as their cytotoxic activity using brine shrimps (Artemia salina) bioassay (Meyer et al., 1982). Saponins 1e4 showed 58%, 42%, 60% and 84% of hemolysis at concentration of 1 mg/ml, respectively. None of them showed cytotoxic activity on brine shrimps at the same concentrations.
4. Chemotaxonomic significance The genus Cephalaria is a member of Dipsacaceae, a taxonomically complex family (Verlaque, 1984). All the isolated iridoides were found for the first time in C. pastricensis. Sweroside, loganin and loganic acid characterize the family Dipsacaceae and could be of value as chemotaxonomic markers of the family (Jensen et al., 1979; Tomita and Mouri, 1996; Perdetzoglou et al., 2000; Horn et al., 2001; Tomassini et al., 2004). Saponins 1e4 were found for the first time in C. pastricensis. With the exception of dipsacoside B (2) this is the first report of their presence in Dipsacaceae family.
D. GoC evac et al. / Biochemical Systematics and Ecology 34 (2006) 890e893
892
O
OR2
R1O CH2OH
1 2 3 4
R1=
-D-Xyl(1-3)- -L-Rha(1-2)- -L-Ara-
R2=
-D-Glc-
R1=
-L-Rha(1-2)- -L-Ara-
R2=
-D-Glc(1-6)- -D-Glc-
R1=
-D-Xyl(1-3)- -L-Rha(1-2)- -L-Ara-
R2=
-D-Glc(1-6)- -D-Glc-
R1=
-D-Glc(1-3)- -L-Rha(1-2)- -L-Ara-
R2=
-D-Glc(1-6)- -D-Glc-
O
O
O
H
O
O
H
H
HO
HO O
O
H
O
H OGlc
OGlc
5
OH
6
OGlc
7
References Alankus-Caliskan, O., Anil, H., 1994. J. Nat. Prod. 57, 1001. Alankus-Caliskan, O., Anil, H., 1995. Phytochemistry 38, 1493. Aliev, A.M., Movsumov, I.S., 1975. Khim. Prir. Soedin. 11, 667. Bianco, A., Passacantilli, P., Polidori, G., Nicoletti, M., Messana, I., 1983. Gazz. Chim. Ital. 113, 829. Bouillant, M.L., Chopin, J., Plouvier, V., 1972. Phytochemistry 11, 1858. Chapelle, J.P., 1976. Planta Med. 29, 268. Diklic´, N., 1973. Dipsacaceae. In: Josifovic´, M. (Ed.), Flore de la Republique Socialiste de Serbie V. Academie Serbe des Sciences et des Arts, Belgrade, pp. 547e548. GoCevac, D., Vajs, V., Menkovic´, N., Tesˇevic´, V., Janac´kovic´, P., Milosavljevic´, S., 2004. J. Serb. Chem. Soc. 69, 883. Horn, M.M., Drewes, S.E., Brown, N.J., Munro, O.Q., Meyer, J.J.M., Mathekga, A.D.M., 2001. Phytochemistry 57, 51. Jensen, S.R., Lyse-Petersen, S.E., Nielsen, B.J., 1979. Phytochemistry 18, 273. Kirmizigul, S., Anil, H., 1994. Phytochemistry 36, 1555. Kirmizigul, S., Anil, H., 2002. Turk. J. Chem. 26, 947. Kirmizigul, S., Anil, H., Rose, M., 1996. Phytochemistry 39, 1171. Kirmizigul, S., Rose, M., 1997. Planta Med. 63, 51. Kong, J., Li, X., Wei, B., Yang, C., 1993. Phytochemistry 33, 427.
D. GoC evac et al. / Biochemical Systematics and Ecology 34 (2006) 890e893
893
Lemmich, E., Cornett, C., Jørstian, C., Knudsen, A.D., Thiilborg, A., Furu, P., Olsen, C., Salih, A., 1995. Phytochemistry 39, 63. Meyer, B.N., Ferrigni, N.R., Putnam, J.E., Jacobsen, L.B., Nichols, D.E., McLaughlin, J.L., 1982. Planta Med. 45, 31. Movsumov, I.S., Aliev, A.M., 1975. Khim. Prir. Soedin. 11, 804. Movsumov, I.S., Aliev, A.M., Kondratenko, E.S., Abubakirov, N.K., 1975. Khim. Prir. Soedin. 11, 519. Mukhamedziev, M.M., Alimbaeva, P.K., Gorovits, T.T., Abubakirov, N.K., 1971. Khim. Prir. Soedin. 7, 153. Pasi, S., Aligiannis, N., Skaltsounis, A., Chinou, I., 2002. Nat. Prod. Lett. 16, 365. Penders, A., Delaude, C., 1994. Carbohydr. Res. 263, 79. Perdetzoglou, D.K., Efthimiopoulos, C., Harvala, C., 2000. Plant Biosys. 134, 67. Tomassini, L., Foddai, S., Nicoletti, M., 2004. Biochem. Syst. Ecol. 32, 1083. Tomita, H., Mouri, Y., 1996. Phytochemistry 42, 239. Verlaque, R., 1984. A biosystematic and phylogenetic study of the Dipsacaceae. In: Grant, W. (Ed.), Plant Biosystematics. Academic Press, Toronto, pp. 307e320. Zemtsova, G.N., Bandyukova, V.A., 1970. Khim. Prir. Soedin. 6, 630. Zviadadze, L.D., Dekanosidze, G.E., Kemertelidze, E.P., 1976. Khim. Prir. Soedin. 13, 423. Zviadadze, L.D., Dekanosidze, G.E., Kemertelidze, E.P., 1980. Khim. Prir. Soedin. 3, 423.