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Flavonoids from Physalis minima
C031-9422188 %300+000 N” 1988 Pergamon Press plc
Phytochemstry. Vol 27, No 11, pp 3708 3709, 198X Prmted I” Great Bntam
FLAVONOIDS
FROM
PH YSALZS MZNIMA
NG ANG SER Department of Chemistry, Natmnal Unlverslty of Smgapore, (Reterwd
VI reomd
form
19 April 1988)
Key Word Index --Phq’sulr~ mmma. Solanaceae, X6.7-trlmethoxyflavone,
Abstract-5-Methoxy-6,7-methylenedloxyflavone compound, 5,6,7-trlmethoxyflavone
was Isolated
Kent Rkdge, Smgapore
from
Smethoxy-6,7-methylenedloxyflavone
Physalls
mmrna
INTRODUCTION
together
with
the known
OMe
Physalzs mmzma IS a common weed of vegetable farms m Singapore and Malaysia In studying the chemical constituents of the plant collected locally, two flavonold substances, 5,6,7-trrmethoxyflavone (1) and 5-methoxy6,7-methylenedloxyflavone (2) were isolated from the plant extract 5,6,7-Trlmethoxyflavone has been reported from a number of different plants [l- 41 while 5-methoxy6,7-methylenedloxyflavone has not been reported prev1ously
/O W
m/z
194
EXPERIMENTAL
RESULTS AYD DISCUSSIO’V Compound 1 was Identified as 5,6,7-tnmethoxyflavone by comparison of spectral data with literature values [lG4] The molecular formula of compound 2 was established as C,,H120, (M’ 296 0685, reyun-ed 296.06847) by high resolution mass spectrometry The IR C=O), UV (215 6, 270 8, 307 0 nm) and (1645 cm-‘, ‘HNMR (66 73, s, lH, C-3 hydrogen) spectra indicated that It was a flavone. The presence of a methylenedloxy group was suggested by the presence of a 2H slgnal at 66.05 (s) m its ‘H NMR and a 13C slgnal at 6 102 12 (t) m its 13CNMR spectra This was further supported by a posltlve Hansen [S] colour reaction for a methylenedloxy group given by 2 The ‘H NMR signals at h7 7 -7 9 (2H, m) and 7.4-7 6 (3H, m) suggested that there was no B rmg substltuents so that the methylenedloxy group must be attached to the A rmg of the flavonold molecule. Examination of Its mass spectrum indicated the usual fragmentation pattern for a flavone [6, 71 A retro-Dlels-Alder reaction of 2 would produce a m/z 194 peak corresponding to the followmg radical Ion The base peak at m/z 166 was attributed to the loss of a CO unit from the above radical ion These were consistent with the suggestion that the methylenedloxy was attached to the A rmg. Furthermore, by comparison with the ‘H NMR of 5,2’-dlmethoxy-6,7-methylenedloxyisofavone [S, 93, the singlet slgnal at 86 64 (7, lH, C-8 proton) indicated that the methylenedloxy group was at C-6 and C-7 Thus, 2 IS ldentlfied as 5-methoxy-6,7methylenedloxyflavone
Mps were uncorr UV spectra were recorded I~I 95% EtOH, ‘H NMR and 13CNMR spectra were recorded m CDCI, usmg TMS as mt standard and mass spectra were recorded by direct inlet at 70eV with VG Micromass 7035 Kleselgel 60 (Merck) was used for CC and Kleselgel Cr. HF254+3hh for TLC P/ant materral Frultmg plant\ of Physal~c mmw~o (H N Rldley, The Floru oJ Malu~ Pemnwlu) were collected m Smgapore, July 1985 and ldentlfied by Professor H Keng Voucher specimen was deposited at the herbarium of the Natlonal Umversity of Singapore lsohon The chopped aerml parts of freshly collected whole plants (8OOg) were extracted with MeOH m two 2 1 round bottom flasks for 2 days with occdslonal shaking The MeOH extract was filtered and the plant material re-extracted with fresh MeOH x 2 The combined MeOH extract was cone to ca SO ml and the coned soln extracted with CHCI, Removal ofthe CHCI, gave 30 g of restdue, Hhlch was washed wtth hexane to remove most of the dark green material It was then separated mto fractions using CC with heuane, hexane-Et,0 and Et,0 as eluents The fraction contammg the flavones was then run on TLC (C,H,-EtOAc 4 I) to give 1 and 2 which were recryst from CH,Cl,-MeOH Compound 1 has identified as 5,6.7trlmethoxyflavone by comparison of Its mp. UV. IR, ‘H NMR and MS with literature values [l-4] Compound 2, S-methoxy6,7-methylenedloxyflavone (60 mg) mp 278-280 . high resolution MS M’ =296 0685, C, ,H, ,O, requires 296 06847. LJV I.?,“,” nm 215 6. 270 8, 307 0 (no changes were observed when NaOMe, AlCI,, AlCI,-HCI, NaOAc and NaOAc--H,BO, were ‘H NMR added), IR Y::: cm -’ 1645 (C-O). 920 (GCH,m0).
3708
3709
Short Reports (CDCI,) 67 70-7 90 (m, 2H), 7 40-7 60 (m, 3H), 6 73 (s, IH), 6 64 (s, 1W, 6 05 (s, 2H), 4 12 (s, 3H), ‘jC NMR (CDCI,): 6 I 77 5 (s, C=Q, C-4), 160 8 (s, C-2 or C-9), 154 7 (s, c-2 or C-9), I 529 (s, c5), 141 4(s, C-7), 134 9 (s, C-6), 131 4 (s, C-l’), 131.2 (d, C-4’), 128 9 (d, C-2’ or C-6’), 125 9 (d, C-3’or C-5’), 112 9 (s, C-lo), 108.3(d, C31,1021 (t, O-W-O), 93 2 (d, C-S), 61 1 (q, OMe), EIMS m/z (rel IntensW) 296 (M + 34 I), 268 (55 5, M-co), 250 (54 s), 222 (lO6)> 237 (106). l94(10 l), 166 (100), 164(967). 136 (339), 105 (17 S), 102 (62 1)
Acknowledgements-1 wish to thank Professor H Keng for ldentlfymg the plant and the Natlonal Umverslty of Smgapore for financial support.
REFERENCES
1 Hosozawa S, Kato, N and Munakata, K (1972) Phytochemistry 11, 2362 Kutney, J P and Hansson, H W (1971) Phytochemrstry 10, 3298 3 Pmar, M (1973) Phytochemrstry 12, 3014 4 Pamchpol, K. and Waterman, P G (1978) Phytochemtstry 17, 1363 5 Hansen, 0 R. (1953) Acfa Chem Stand 7, 1125 6 Kmgston, D. G I (1971) Tetrahedron 27, 2691 7. (1975) The Flavonolds (Harbone, J B, Mabry, T J. and Mabry, H , eds) Chapman & Hall, London. 8 Gelgert, J , Stermltz, F. R., Johnson, G., Maag, D D and Johnson, D K. (1973) Tetrahedron 29,2703. 9. ChlJl, H., Arakawa, Y, Ueda, S, Kuroda, M. and Igawa, M (1986) Phytochemzstry 25, 281
Phytockmrstry, Vol. 27, No 11, pp 3709-3711, 1988 0
Pnnted In Great Bntam
0031-9422/88 $3.00+0 00 1988 PergamonPress plc
%HYDROXYTRICETIN ‘I-GLUCURONIDE, A /3-GLUCURONIDASE INHIBITOR FROM SCOPARIA DULCIS MASARU
KAWASAKI,
TOSHIMITSU HAYASHI,
MUNEHISA ARISAWA,
NAOKATA
MORITA
and Lurs H BERGANZA*
Faculty of Pharmaceutical Sciences, Toyama Me&al and Pharmaceutical Umverslty, 2630 Sugltam, Toyama 930-01, Japan, *Facultad de Clenclas Qmmlcas, Umversidad National de Asun&n Casllla de Correo 1055, AsuncGn, Paraguay (Received m revned form 12 Aprd 1988)
Key Word Index--Scoparla d&s, romde, b-glucuromdase mhlbitor
Scrophulanaceae, flavonmds, 5,7,8,3’,4’,5’-hexahydroxyflavone
7-0-/3-D-ghCU-
Abstract-A new flavone glycoslde has been Isolated from Scoparia dulcis together with 11 known compounds. The new glycoside was determmed as 5,7,8,3’,4’,5’-hexahydroxyflavone glucuromde by spectral analysis. The new glycoside and isovitexm showed inhlbltory actlvlty agamst fi-glucuromdase
INTRODUCTION
Scoparza dulcrs L is a perennial herb which has been used for the treatment of stomach disease and hepatosis m Paraguay, as a cure for hypertension in Taiwan [ 1,2] and for toothache, blennorrhagia and stomach problems m India [3]. From Indian S. d&is, an antldiabetlc compound named amellm was Isolated by Nath [4] Earlier phytochemlcal studies on this medicmal plant resulted m isolation of hexacosanol, D-manmtol, sitosterol [3] and 6-methoxybenzoxazolmone [S] as well as trlterpenoids [5, 63 and flavonolds [7] Previously, we reported the lsolatlon and structural elucidation of five new diterpene acids from the 70% ethanohc extract of a plant collected m Paraguay [8-lo] In a continuation of this work, we have exammed the water-soluble fraction which showed
mild mhlbltory activity against fi-glucuronidase. This paper deals with isolation of flavonoids from this fraction and their mhlbitory activity against p-glucuromdase.
RESULTS AND DISCUSSION
Eleven flavonoids (l-11) and a phenylpropanoid (12) were isolated from the water-soluble fraction of a 70% ethanolic extract of S. dulcis. The compounds, l-10, 12, were ldentlfied as apigenin (l), scutellarein (2), luteolm (3), vlcenm-2 (4), lmarm (5), vitexm (6), lsovltexin (7), scutellarm (S), scutellarm methyl ester (9), luteolm 7-glucoside (10) and p-coumarlc acid (12) by direct comparison of their physical and spectroscopic properties (mp, IR, LJV, ‘H NMR and 13C NMR) with those of authentic samples,
Phytochemstry. Vol 27, No 11, pp 3708 3709, 198X Prmted I” Great Bntam
FLAVONOIDS
FROM
PH YSALZS MZNIMA
NG ANG SER Department of Chemistry, Natmnal Unlverslty of Smgapore, (Reterwd
VI reomd
form
19 April 1988)
Key Word Index --Phq’sulr~ mmma. Solanaceae, X6.7-trlmethoxyflavone,
Abstract-5-Methoxy-6,7-methylenedloxyflavone compound, 5,6,7-trlmethoxyflavone
was Isolated
Kent Rkdge, Smgapore
from
Smethoxy-6,7-methylenedloxyflavone
Physalls
mmrna
INTRODUCTION
together
with
the known
OMe
Physalzs mmzma IS a common weed of vegetable farms m Singapore and Malaysia In studying the chemical constituents of the plant collected locally, two flavonold substances, 5,6,7-trrmethoxyflavone (1) and 5-methoxy6,7-methylenedloxyflavone (2) were isolated from the plant extract 5,6,7-Trlmethoxyflavone has been reported from a number of different plants [l- 41 while 5-methoxy6,7-methylenedloxyflavone has not been reported prev1ously
/O W
m/z
194
EXPERIMENTAL
RESULTS AYD DISCUSSIO’V Compound 1 was Identified as 5,6,7-tnmethoxyflavone by comparison of spectral data with literature values [lG4] The molecular formula of compound 2 was established as C,,H120, (M’ 296 0685, reyun-ed 296.06847) by high resolution mass spectrometry The IR C=O), UV (215 6, 270 8, 307 0 nm) and (1645 cm-‘, ‘HNMR (66 73, s, lH, C-3 hydrogen) spectra indicated that It was a flavone. The presence of a methylenedloxy group was suggested by the presence of a 2H slgnal at 66.05 (s) m its ‘H NMR and a 13C slgnal at 6 102 12 (t) m its 13CNMR spectra This was further supported by a posltlve Hansen [S] colour reaction for a methylenedloxy group given by 2 The ‘H NMR signals at h7 7 -7 9 (2H, m) and 7.4-7 6 (3H, m) suggested that there was no B rmg substltuents so that the methylenedloxy group must be attached to the A rmg of the flavonold molecule. Examination of Its mass spectrum indicated the usual fragmentation pattern for a flavone [6, 71 A retro-Dlels-Alder reaction of 2 would produce a m/z 194 peak corresponding to the followmg radical Ion The base peak at m/z 166 was attributed to the loss of a CO unit from the above radical ion These were consistent with the suggestion that the methylenedloxy was attached to the A rmg. Furthermore, by comparison with the ‘H NMR of 5,2’-dlmethoxy-6,7-methylenedloxyisofavone [S, 93, the singlet slgnal at 86 64 (7, lH, C-8 proton) indicated that the methylenedloxy group was at C-6 and C-7 Thus, 2 IS ldentlfied as 5-methoxy-6,7methylenedloxyflavone
Mps were uncorr UV spectra were recorded I~I 95% EtOH, ‘H NMR and 13CNMR spectra were recorded m CDCI, usmg TMS as mt standard and mass spectra were recorded by direct inlet at 70eV with VG Micromass 7035 Kleselgel 60 (Merck) was used for CC and Kleselgel Cr. HF254+3hh for TLC P/ant materral Frultmg plant\ of Physal~c mmw~o (H N Rldley, The Floru oJ Malu~ Pemnwlu) were collected m Smgapore, July 1985 and ldentlfied by Professor H Keng Voucher specimen was deposited at the herbarium of the Natlonal Umversity of Singapore lsohon The chopped aerml parts of freshly collected whole plants (8OOg) were extracted with MeOH m two 2 1 round bottom flasks for 2 days with occdslonal shaking The MeOH extract was filtered and the plant material re-extracted with fresh MeOH x 2 The combined MeOH extract was cone to ca SO ml and the coned soln extracted with CHCI, Removal ofthe CHCI, gave 30 g of restdue, Hhlch was washed wtth hexane to remove most of the dark green material It was then separated mto fractions using CC with heuane, hexane-Et,0 and Et,0 as eluents The fraction contammg the flavones was then run on TLC (C,H,-EtOAc 4 I) to give 1 and 2 which were recryst from CH,Cl,-MeOH Compound 1 has identified as 5,6.7trlmethoxyflavone by comparison of Its mp. UV. IR, ‘H NMR and MS with literature values [l-4] Compound 2, S-methoxy6,7-methylenedloxyflavone (60 mg) mp 278-280 . high resolution MS M’ =296 0685, C, ,H, ,O, requires 296 06847. LJV I.?,“,” nm 215 6. 270 8, 307 0 (no changes were observed when NaOMe, AlCI,, AlCI,-HCI, NaOAc and NaOAc--H,BO, were ‘H NMR added), IR Y::: cm -’ 1645 (C-O). 920 (GCH,m0).
3708
3709
Short Reports (CDCI,) 67 70-7 90 (m, 2H), 7 40-7 60 (m, 3H), 6 73 (s, IH), 6 64 (s, 1W, 6 05 (s, 2H), 4 12 (s, 3H), ‘jC NMR (CDCI,): 6 I 77 5 (s, C=Q, C-4), 160 8 (s, C-2 or C-9), 154 7 (s, c-2 or C-9), I 529 (s, c5), 141 4(s, C-7), 134 9 (s, C-6), 131 4 (s, C-l’), 131.2 (d, C-4’), 128 9 (d, C-2’ or C-6’), 125 9 (d, C-3’or C-5’), 112 9 (s, C-lo), 108.3(d, C31,1021 (t, O-W-O), 93 2 (d, C-S), 61 1 (q, OMe), EIMS m/z (rel IntensW) 296 (M + 34 I), 268 (55 5, M-co), 250 (54 s), 222 (lO6)> 237 (106). l94(10 l), 166 (100), 164(967). 136 (339), 105 (17 S), 102 (62 1)
Acknowledgements-1 wish to thank Professor H Keng for ldentlfymg the plant and the Natlonal Umverslty of Smgapore for financial support.
REFERENCES
1 Hosozawa S, Kato, N and Munakata, K (1972) Phytochemistry 11, 2362 Kutney, J P and Hansson, H W (1971) Phytochemrstry 10, 3298 3 Pmar, M (1973) Phytochemrstry 12, 3014 4 Pamchpol, K. and Waterman, P G (1978) Phytochemtstry 17, 1363 5 Hansen, 0 R. (1953) Acfa Chem Stand 7, 1125 6 Kmgston, D. G I (1971) Tetrahedron 27, 2691 7. (1975) The Flavonolds (Harbone, J B, Mabry, T J. and Mabry, H , eds) Chapman & Hall, London. 8 Gelgert, J , Stermltz, F. R., Johnson, G., Maag, D D and Johnson, D K. (1973) Tetrahedron 29,2703. 9. ChlJl, H., Arakawa, Y, Ueda, S, Kuroda, M. and Igawa, M (1986) Phytochemzstry 25, 281
Phytockmrstry, Vol. 27, No 11, pp 3709-3711, 1988 0
Pnnted In Great Bntam
0031-9422/88 $3.00+0 00 1988 PergamonPress plc
%HYDROXYTRICETIN ‘I-GLUCURONIDE, A /3-GLUCURONIDASE INHIBITOR FROM SCOPARIA DULCIS MASARU
KAWASAKI,
TOSHIMITSU HAYASHI,
MUNEHISA ARISAWA,
NAOKATA
MORITA
and Lurs H BERGANZA*
Faculty of Pharmaceutical Sciences, Toyama Me&al and Pharmaceutical Umverslty, 2630 Sugltam, Toyama 930-01, Japan, *Facultad de Clenclas Qmmlcas, Umversidad National de Asun&n Casllla de Correo 1055, AsuncGn, Paraguay (Received m revned form 12 Aprd 1988)
Key Word Index--Scoparla d&s, romde, b-glucuromdase mhlbitor
Scrophulanaceae, flavonmds, 5,7,8,3’,4’,5’-hexahydroxyflavone
7-0-/3-D-ghCU-
Abstract-A new flavone glycoslde has been Isolated from Scoparia dulcis together with 11 known compounds. The new glycoside was determmed as 5,7,8,3’,4’,5’-hexahydroxyflavone glucuromde by spectral analysis. The new glycoside and isovitexm showed inhlbltory actlvlty agamst fi-glucuromdase
INTRODUCTION
Scoparza dulcrs L is a perennial herb which has been used for the treatment of stomach disease and hepatosis m Paraguay, as a cure for hypertension in Taiwan [ 1,2] and for toothache, blennorrhagia and stomach problems m India [3]. From Indian S. d&is, an antldiabetlc compound named amellm was Isolated by Nath [4] Earlier phytochemlcal studies on this medicmal plant resulted m isolation of hexacosanol, D-manmtol, sitosterol [3] and 6-methoxybenzoxazolmone [S] as well as trlterpenoids [5, 63 and flavonolds [7] Previously, we reported the lsolatlon and structural elucidation of five new diterpene acids from the 70% ethanohc extract of a plant collected m Paraguay [8-lo] In a continuation of this work, we have exammed the water-soluble fraction which showed
mild mhlbltory activity against fi-glucuronidase. This paper deals with isolation of flavonoids from this fraction and their mhlbitory activity against p-glucuromdase.
RESULTS AND DISCUSSION
Eleven flavonoids (l-11) and a phenylpropanoid (12) were isolated from the water-soluble fraction of a 70% ethanolic extract of S. dulcis. The compounds, l-10, 12, were ldentlfied as apigenin (l), scutellarein (2), luteolm (3), vlcenm-2 (4), lmarm (5), vitexm (6), lsovltexin (7), scutellarm (S), scutellarm methyl ester (9), luteolm 7-glucoside (10) and p-coumarlc acid (12) by direct comparison of their physical and spectroscopic properties (mp, IR, LJV, ‘H NMR and 13C NMR) with those of authentic samples,