146
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Cardona, M.L., Fernández, I., Pedro, J.R., Serrano, A., 1990. Xanthones from
Hypericum reflexum. Phytochemistry 29, 3003–3006.
Cheminat, A., Zawatzky, R., Becker, H., Brouillard, R., 1988. Caffeoyl conjugates from
Echinacea species: structure and biological activity. Phytochemistry 27, 2787–
2794.
Chen, M.-T., Chen, C.-M., 1985. Xanthones from Hypericum sampsonii. Heterocycles
23, 2543–2548.
Corthout, J., Pieters, L., Claeys, M., Vanden Berghe, D., Vlietinck, A., 1992. Plant
antiviral agents. 9. Antiviral caffeoyl esters from Spondias mombin.
Phytochemistry 31, 1979–1981.
Faizi, S., Zikr-ur-Rehman, S., Ali, M., Naz, A., 2006. Temperature and solvent
dependent NMR studies on magniferin and complete NMR spectral assignments
of its acyl and methyl derivatives. Magn. Reson. Chem. 44, 838–844.
Frahm, A.W., Chaudhuri, R.K., 1979. 13C NMR spectroscopy of substituted
xanthones-II: 13C NMR spectral study of polyhydroxy xanthones. Tetrahedron
35, 2035–2038.
J = 2.0 Hz), 6.08 (1H, ddd, J = 17.2, 10.8, 5.2 Hz), 5.45 (1H, dd,
J = 17.2, 1.2 Hz), 5.29 (1H, dd, J = 10.8, 1.2 Hz), 4.67 (2H, d,
J = 5.2 Hz), 1.93 (3H, s). 7-Dimethylallyloxy-5-hydroxy-3-meth-
ylchromone (6f): (26 mg, 65% yield). Amorphous powder. HRE-
SIMS: m/z 283.0927, [M+Na]+ (calcd for C15H16O4Na, 283.0946);
1H NMR (acetone-d6): dH 12.83 (1H, s), 8.03 (1H, d, J = 0.8 Hz),
6.47 (1H, d, J = 1.2 Hz), 6.31 (1H, d, J = 1.2 Hz), 5.47 (1H, td, J = 6.8,
0.8 Hz), 4.66 (2H, d, J = 6.8 Hz), 1.94 (3H, d, J = 0.8 Hz), 1.79 (3H,
d, J = 6.8 Hz), 1.77 (3H, s).
4.14. Anti-Helicobacter pylori activity
The disc-diffusion method was used to screen compounds
against H. pylori ATCC43504 and H. pylori SS-1 as described previ-
ously (Ochi et al., 2005).
Foo, L.Y., Lu, Y., Molan, A.L., Woodfield, D.R., MacNabb, W.C., 2000. The phenols and
prodelphinidins of white clover flowers. Phytochemistry 54, 539–548.
Hashida, W., Tanaka, N., Takaishi, Y., 2007. Prenylated xanthones from Hypericum
ascyron. J. Nat. Med. 61, 371–374.
Jürgenliemk, G., Nahrstedt, A., 2002. Phenolic compounds from Hypericum
perforatum. Planta Med. 68, 88–91.
4.15. Cell Lines and Cell Culture
Lee, E.H., Kim, H.J., Song, Y.S., Jin, C., Lee, K.-T., Cho, J., Lee, Y.S., 2003. Constituents of
the stems and fruits of Opuntia ficus-indica var. Saboten. Arch. Pharmacal. Res.
26, 1018–1023.
KB (human epidermoid carcinoma of the nasopharynx), MCF-7
(breast carcinoma), COLO205 (colon carcinoma), K-562 (leukemia),
and K562/Adr (multidrug-resistant human erythromyelogenous
leukemia) cells were obtained from the Cell Resource Center for
Biomedical Research (Tohoku University). Multidrug-resistant hu-
man epidermoid carcinoma KB-C2 cells were kindly provided by
Prof. Shin-ichi Akiyama (Kagoshima University, Japan). KB cells
were cultured in Dulbecco’s modified Eagles medium (DMEM)
with 10% fetal bovine serum (FBS). KB-C2 cells were maintained
Markham, K.R., Ternai, B., 1976. 13C NMR of flavonoids-II: flavonoids other then
flavone and flavonol aglycones. Tetrahedron 32, 2607–2612.
Markham, K.R., Ternai, B., Stanley, R., Geiger, H., Mabry, T.J., 1978. Carbon-13 NMR
studies of flavonoids-III: naturally occurring flavonoid glycosides and their
acylated derivatives. Tetrahedron 34, 1389–1397.
Matsuhisa, M., Shikishima, Y., Takaishi, Y., Honda, G., Ito, M., Takeda, Y., Shibata, H.,
Higuti, T., Kodzhimatov, O.K., Ashurmetov, O., 2002. Benzoylphloroglucinol
derivatives from Hypericum scabrum. J. Nat. Prod. 65, 290–294.
Morikawa, H., Kasai, R., Otsuka, H., Hirata, E., Shinzato, T., Aramoto, M., Takeda, Y.,
2004. Terpenic and phenolic glycosides from leaves of Breynia officinalis Hemsl.
Chem. Pharm. Bull. 52, 1086–1090.
Nedialkov, P.T., Kitanov, G.M., 2002. Two benzophenone O-arabinosides and a
chromone from Hypericum annulatum. Phytochemistry 59, 867–871.
Ochi, T., Shibata, H., Higuti, T., Kodama, K., Kusumi, T., Takaishi, Y., 2005. Anti-
Helicobacter pylori compounds from Santalum album. J. Nat. Prod. 68, 819–824.
Peng, L.-Y., Mei, S.-X., Jiang, B., Zhou, H., Sun, H.-D., 2000. Constituents from Lonicera
japonica. Fitoterapia 71, 713–715.
in DMEM medium in the presence of 10% FBS and 2 lg/ml colchi-
cine. MCF-7, COLO205, and K-562 cells were cultured in RPMI1640
supplemented with 10% FBS. K-562/Adr (doxorubicin-resistant
K562 cell line) cells were cultured in RPMI1640 medium contain-
ing 10% FBS and 0.5 lM doxorubicin. All cells were incubated at
Piao, M.S., Kim, M.-R., Lee, D.G., Park, Y., Hahm, K.-S., Moon, Y.-H., Woo, E.-R., 2003.
Antioxidative constituents from Buddlea officinalis. Arch. Pharmacal. Res. 26,
453–457.
Pinaros, A., 2005. A survey on the Hypericum genus: secondary metabolites and
bioactivity. Stud. Nat. Chem. 30, 603–634.
37ꢀC in a humidified atmosphere with 5% CO2 to 95% air.
4.16. Cytotoxicity Assays
Robeson, D.J., Ingham, J.L., Harborne, B., 1980. Identification of two chromone
phytoalexins in the sweet pea, Lathyrus odoratus. Phytochemistry 19, 2171–
2173.
Cells in exponential growth were trypsinized, dispersed in a sin-
gle cell suspension, and dispensed in 100 ll volumes into 96-well
plates. For each assay, 1 ꢁ 104 cells/well for K562 and K562/Adr,
Runbero-Sanchez, A., Vazquez, P., 1991. Quinic acid esters from Isertia haenkeana.
Phytochemistry 30, 311–313.
Sakushima, A., Coskun, M., Maoka, T., 1995. Hydroxybenzoic acids from Boreava
orientalis. Phytochemistry 40, 257–261.
5 ꢁ 103 cells /well for KB and KB-C2, or 5 ꢁ 103 cells for MCF-7,
and COLO205 were inoculated in 100 ll medium containing 10%
FBS and incubated for 24 h. Test samples were dissolved in small
amounts of DMSO and diluted in the appropriate culture medium
(final concentration of DMSO <0.5%). After removal of pre-incu-
Satake, T., Kamiya, K., Saiki, Y., Hama, T., Fujimoto, Y., Endang, H., Umar, M., 1999.
Chromone C-glycosides from Baeckea frutescens. Phytochemistry 50, 303–306.
Tanaka, N., Takaishi, Y., Shikishima, Y., Nakanishi, Y., Bastow, K., Lee, K.-H., Honda,
G., Ito, M., Takeda, Y., Kodzhimatov, O.K., Ashurmetov, O., 2004. Prenylated
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1875.
Tanaka, N., Okasaka, M., Ishimaru, Y., Takaishi, Y., Sato, M., Okamoto, M., Oshikawa,
T., Ahmed, S.U., Consentino, L.M., Lee, K.-H., 2005. Biyouyanagin A, an anti-HIV
agent from Hypericum chinense L. var. salicifolium. Org. Lett. 7, 2997–2999.
Tanaka, N., Takaishi, Y., 2006. Xanthones from Hypericum chinense. Phytochemistry
67, 2146–2151.
bated culture medium, 100 ll of medium containing various con-
centrations of each test compound were added and further
incubated for 48 h. Cell proliferation was determined by a 3-(4,5-
dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT)
assay. IC50 values are defined as the concentration of each test
samples that reduced absorbance to 50% of vehicle-treated
controls.
Tanaka, N., Takaishi, Y., 2007. Xanthones from stems of Hypericum chinense. Chem.
Pharm. Bull. 55, 19–21.
Ushiyama, M., Kumagai, S., Furuya, T., 1989. Biotransformation of phenylcarboxylic
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Wu, Q.-L., Wang, S.-P., Du, L.-J., Zhang, S.-M., Yang, J.-S., Xiao, P.-G., 1998. Chromone
glycosides and flavonoids from Hypericum japonicum. Phytochemistry 49,
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Zhang, W., Ha, T.B.T., Chen, W., Kong, D., Li, H., Wang, Y., Isabella, F., 2001. Study on
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