Coumarins from the fruits of Seseli devenyense

Article abstract of DOI:10.1021/np050209w

Eight new coumarins were isolated from the fruits of Seseli devenyense Simonkai. Their structures were established from NMR and mass data and their absolute configurations from chemical degradation correlation reactions. The new structures are the decanoic and dodecanoic esters of (+)-lomatin (3, 4), the decanoates of (+)-cis-khellactone at positions 4′ (5) and 3′ (6) as well as the 2′S epimer of 8-(2,3-dihydroxy-3-methylbutyl)-7- hydroxychromen-2-one (7) named devenyol, its two O-monoglucosides at positions 3′ and 7 named devenyosides A (8) and B (9), and the corresponding 3′- and 7-O-diglucoside named devenyoside C (10). This plant is an interesting example of stereochemical diversity based on biodiversity given that other members of the Apiaceae family produce exclusively the 2′R epimers of compounds 7-9.

Full text of DOI:10.1021/np050209w

J. Nat. Prod. 2005, 68, 1637-1641
1637
Coumarins from the Fruits of Seseli devenyense
Jarek Widelski,†,‡ Eleni Melliou,† Nikolas Fokialakis,† Prokopios Magiatis,† Kazimierz Glowniak,‡ and
Ioanna Chinou*,†
Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, University of Athens,
Panepistimiopolis Zografou, Athens GR-15771, Greece, and Department of Pharmacognosy, Faculty of Pharmacy,
Skubiszewski Medicinal University of Lublin, Lublin, Poland
Received June 11, 2005
Eight new coumarins were isolated from the fruits of Seseli devenyense Simonkai. Their structures were
established from NMR and mass data and their absolute configurations from chemical degradation
correlation reactions. The new structures are the decanoic and dodecanoic esters of (+)-lomatin (3, 4),
the decanoates of (+)-cis-khellactone at positions 4′ (5) and 3′ (6) as well as the 2′S epimer of 8-(2,3-
dihydroxy-3-methylbutyl)-7-hydroxychromen-2-one (7) named devenyol, its two O-monoglucosides at
positions 3′ and 7 named devenyosides A (8) and B (9), and the corresponding 3′- and 7-O-diglucoside
named devenyoside C (10). This plant is an interesting example of stereochemical diversity based on
biodiversity given that other members of the Apiaceae family produce exclusively the 2′R epimers of
compounds 7-9.
Coumarins represent a category of heterocyclic natural
compounds characterized by extensive chemodiversity, with
more than 1800 reported structures and a variety of
pharmacological activities.1 One very interesting subclass
of this category is linear or angular pyranocoumarins that
possess antiproliferative,2 antiviral,3 or antibacterial4 ac-
tivities.
The genus Seseli (Apiaceae) is a well-known source of
the aforementioned subclass of coumarins, and it contains
numerous species that have been used in folk medicine
since ancient times.5
Having as target the investigation of the chemodiversity
of the natural pyranocoumarins and their related metabo-
lites, we studied the fruits of Seseli devenyense Simonkai,
a plant widespread in Eastern and Central Europe6 that
has not been previously studied. The plants were cultivated
in the Botanical Garden of the Department of Pharmacog-
nosy of the Medical University of Lublin, Poland.
Results and Discussion
Fourteen coumarins were isolated by fractionation of the
MeOH extract of the ripe fruits of S. devenyense. Four
known coumarins were identified, (+)-cis-khellactone7 and
its derivatives laserpitine,8 3′-O-angeloyl-cis-khellactone,9
and praeroside II.10 The MeOH extract also contained a
variety of medium-chain (C6-C12) aliphatic esters of (+)-
lomatin and (+)-cis-khellactone. The hexanoate and oc-
tanoate of lomatin (1, 2) had been previously reported as
The empirical formula of compound 3 was established
by accurate mass measurement as C24H32O5. The UV
spectrum was suggestive of a 7-oxygenated coumarin
chromophore.13 The 1H NMR spectrum displayed two pairs
of doublets at 6.24 and 7.63 ppm (J ) 9.5 Hz) and at 6.79
and 7.26 ppm (J ) 8.6 Hz) typical for H-3, H-4, H-6, and
H-5 of a 7,8-disubstituted coumarin.2,13 Other high-field
signals accounted for a -CH2CH- system (5.13, t, J ) 5.0
Hz, H-3′, 3.19, dd, J ) 15.1, 5.0 Hz, H-4′a, 2.97, dd, J )
15.1, 5.0 Hz, H-4′b), two methyls (1.37, 1.34 ppm), and a
set of signals corresponding to a linear aliphatic ester. The
NMR profile of 3 was very similar to that of previously
reported lomatin esters.11 Indeed, the carbonyl of the
aliphatic ester showed a cross-peak in the HMBC spectrum
with the proton at 5.13 ppm (H-3′), while this proton was
constituents of S. gummiferum,11 while 2 had also been
reported from Libanotis lehmannae.12 The decanoate and
dodecanoate of lomatin (3, 4) as well as the decanoate of
(+)-cis-khellactone at positions 4′ (5) and 3′ (6) are reported
herein for the first time. Additionally, a new coumarin
aglycone, the 2′S epimer of 8-(2,3-dihydroxy-3-methylbu-
tyl)-7-hydroxychromen-2-one (7), and three new glucosides,
the O-monoglucosides at positions 3′ (8) and 7 (9) and the
corresponding 3′- and 7-O-diglucoside (10), were isolated
and identified.
* To whom correspondence should be addressed. Fax: +30210 7274115.
Tel: +30210 7274595. E-mail: ichinou@pharm.uoa.gr.
† University of Athens.
‡ University of Lublin.
10.1021/np050209w CCC: $30.25
© 2005 American Chemical Society and American Society of Pharmacognosy
Published on Web 10/19/2005

1638 Journal of Natural Products, 2005, Vol. 68, No. 11
Widelski et al.
correlated with the geminal methyls of the pyrano ring.
The length of the aliphatic ester was deduced by the
alkaline hydrolysis of 3 that afforded decanoic acid, which
was identified by GC-MS. Thus, the structure of compound
3 was established as decanoyllomatin. Alkaline hydrolysis
of 3 also afforded (+)-lomatin, which was identified by
NMR, MS, and [R]D data,14,15 revealing that the configu-
ration at C-3′ of 3 is R. The absolute configuration of the
prepared lomatin was unequivocally confirmed using Mosh-
er ester methodology as previously described.16
In a similar fashion, compound 4 was identified as
dodecanoyllomatin. The UV, IR, and NMR spectra were
almost identical with 3, but the HRMS data showed that
this compound contained an additional C2H4 group obvi-
ously corresponding to two more methylenes on the ali-
phatic side chain. Indeed hydrolysis of 4 afforded (+)-
lomatin and dodecanoic acid and confirmed the 3′R
configuration.
chromic shift with NaOAc). The 1H NMR and 13C NMR
spectra showed a clear resemblance to 7, but in the case of
8 an additional set of peaks corresponding to a glucose
moiety (anomeric proton at 4.58 ppm, J ) 7.7 Hz) was
observed. The identification of â-D-glucose was based on
(1) the enzymatic hydrolysis of 8 by â-D-glucosidase and
(2) the presence of characteristic coupling constants of H-1′′,
2′′, 3′′, and 4′′, which were more clearly observed in the
peracetylated derivative 8a. The enzymatic hydrolysis
afforded as aglycone compound 7, identified by NMR, MS,
and specific rotation data, suggesting that 8 is a glucoside
derivative of 7 with the same configuration (S) at position
2′. The site of attachment of glucose was revealed by the
HMBC data, which showed a 3J correlation of the anomeric
proton to the quaternary oxygenated carbon (C-3′) of the
isoprenyl side chain. Consequently, 8 is the 2′S epimer of
tortuoside,17 named devenyoside A.
The HRMS analysis of compound 9 gave the same
molecular formula as 8. An important difference with 8 was
observed in the UV spectrum, where the addition of NaOAc
did not induce a bathochromic shift, suggesting that the
hydroxyl at position 7 was substituted. As in the case of 8
the 1H NMR and 13C NMR spectra showed the typical
signals of a simple 7,8-bisubstituted coumarin and a
glucose moiety. Some important differences in the 1H NMR
spectrum as compared with 8 were the downfield shift of
the H-6 (+0.54 ppm) and of the anomeric proton (+0.38
ppm), suggesting the attachment of the sugar to the
phenolic hydroxyl. Enzymatic hydrolysis of 9, as in the case
of 8, afforded as aglycone compound 7, identified by NMR,
MS, and specific rotation data, suggesting that 9 was also
a glucoside derivative of 7 with the same configuration (S)
at position 2′. The site of attachment of glucose was
identified from the HMBC spectrum, where a 3J correlation
of the anomeric proton with C-7 was observed, confirming
the initial observation of the UV spectrum. Consequently,
9 is the (-)-7-O-â-D-glucopyranoside of 8-[(2S),3-dihydroxy-
3-methylbutyl]-7-hydroxychromen-2-one, named devenyo-
side B. The 2′ epimer of devenyoside B had been previously
reported from Diplolophium buchananii (Apiaceae).18
Finally, compound 10 was shown to have the molecular
formula C26H36O15, an increase of C6H10O5 when compared
with 8 or 9, suggesting the presence of an additional
hexosyl moiety The UV spectrum of 10, as in the case of 9,
suggested that 7-OH was substituted. The NMR spectra
of 10 were similar to those of 8 and 9, but the observation
of two anomeric protons indicated that it was a diglycoside.
The enzymatic hydrolysis of 10, as in the case of 8 and 9,
afforded compound 7, previously identified, suggesting that
10 was a diglucoside derivative of 7 once again possessing
the same configuration (S) at position 2′. The sites of
attachment of the two glucosyl moieties were indicated by
the HMBC spectrum, where the first anomeric proton was
correlated with C-7 and the second anomeric proton with
C-3′ of the isoprenyl side chain. Consequently, 10 is (-)-
3′-O-â-D-glucopyranosyl-8-[(2S),3-dihydroxy-3-methylbutyl]-
7-hydroxychromen-2-one 7-O-â-D-glucopyranoside, named
devenyoside C.
Accurate mass measurement of compound 5 indicated a
molecular formula of C24H32O6. Analysis of the NMR
spectra indicated that it was an esterified derivative of cis-
khellactone. When compared with the spectrum of cis-
khellactone, the NMR spectra of 5 showed additional
resonances for one long linear aliphatic ester (with 10
carbon atoms), while the chemical shift of H-4′ (6.41 ppm,
J ) 4.9 Hz) suggested esterification at this site. In addition,
the HMBC spectrum showed that H-4′ (identified by its
correlations with C-2′, C-8a, and C-7) was also correlated
with the carbonyl of the aliphatic moiety. Acid hydrolysis
in MeOH afforded decanoic acid, identified by GC-MS, (+)-
cis-methylkhellactone (resolved using Mosher ester meth-
odology), and (-)-trans-methylkhellactone as previously
reported for 4′-esters of (+)-cis-khellactone16 and confirming
the absolute configuration as 3′R, 4′R.
Compound 6 had the same molecular formula as 5, with
their UV, IR, and NMR spectra being very similar. The
1
only important difference in the H NMR spectrum was
the chemical shifts of H-3′ and H-4′, suggesting a change
in the position of esterification in comparison with 5.
Indeed the HMBC spectrum showed that the carbonyl of
the decanoyl moiety this time was correlated with H-3′.
Alkaline hydrolysis of 6 gave decanoic acid, identified by
GC-MS, and exclusively (+)-cis-khellactone (identified as
previously) as expected for the case of homobenzylic esters,
confirming the absolute configuration as 3′R, 4′R.
The molecular formula of compound 7 was found to be
C14H16O5 by HRMS. The IR and UV spectra were sugges-
tive of a simple coumarin with a free hydroxyl at position
1
7 (bathochromic shift with NaOAc). The H NMR and 13C
NMR spectra, apart from the typical set of signals corre-
sponding to positions 3, 4, 5, and 6, revealed the presence
of an isoprenyl chain dihydroxylated at positions 2′ and
3′. The NMR data were identical with those described for
tortuoside aglycone17 prepared by hydrolysis of tortuoside
from Seseli tortuosum. The only difference between 7 and
tortuoside aglycone was related to the sign of the specific
rotation. The absolute configuration of that compound had
been established as 2′R on the basis of chemical correlation
reactions. The opposite sign of the optical rotation found
for 7 suggests that it is the 2′ epimer of tortuoside aglycone.
Consequently, compound 7 is 8-[(2S),3-dihydroxy-3-meth-
ylbutyl]-7-hydroxychromen-2-one, for which the name de-
venyol is proposed.
The most interesting aspect of this work is the number
of new coumarins isolated from a single plant. Considering
the large number of coumarins already known,1 this indeed
represents a significant find. It should also be noted that
this plant is an interesting example of stereochemical
diversity based on biodiversity given that other members
of the Apiaceae family produce exclusively the 2R epimers
of compounds 7-9.
Compound 8 was obtained as an amorphous solid with
molecular formula C20H26O10 given by HRMS. The IR and
UV spectra were similar to those of 7, also suggesting a
simple coumarin with a free hydroxyl at position 7 (batho-

Coumarins from the Fruits of Seseli devenyense
Journal of Natural Products, 2005, Vol. 68, No. 11 1639
Experimental Section
fractions of 20 mL each: fractions C1-C20, H2O eluted; C21-
C59, H2O/MeOH (90:10); C60-C85, H2O/MeOH (80:20) eluted;
C86-C130, H2O/MeOH (70:30) eluted. Fractions C60-C65
afforded devenyoside C (10) (10 mg), C78-C84 devenyoside B
(9) (20 mg), C91-C100 devenyoside A (8) (80 mg), and C105-
C115 praeroside II (5 mg).
General Experimental Procedures. Specific rotations
were measured with a Perkin-Elmer 341 polarimeter. UV
spectra were recorded on a Shimadzu-160A spectrophotometer.
The IR spectra were obtained on a Perkin-Elmer Paragon 500
instrument. NMR spectra were recorded at 25 °C on Bruker
DRX 400 and Bruker AC 200 spectrometers [1H (400 MHz)
and 13C (50 MHz)]; chemical shifts are expressed in ppm
downfield to TMS, which was used as internal standard. The
2D NMR experiments (COSY, HMQC, and HMBC) were
performed using standard Bruker microprograms with gradi-
ent pulse sequences. EIMS were determined on an HP-6890
spectrometer, FABMS were obtained using a ZAB HF instru-
ment, HREIMS and HRFABMS were obtained on an AEI MS-
902 mass spectrometer. Liquid chromatography was performed
on columns containing Si gel 60 Merck (40-63 µm). The GC-
MS analyses were carried out using a Hewlett-Packard 6890-
5973 GC-MS system operating in EI mode (equipped with a
HP 5MS 30 m × 0.25 mm, 0.25 µm film thickness capillary
column). He (1 mL/min) was used as carrier gas. The initial
temperature of the column was 60 °C, and then it was heated
to 280 °C at a rate of of 3 °C/min. The identification of
compounds by GC-MS was based on comparison of their
retention indices (RI), obtained using n-alkanes (C9-C25), by
comparison of their EI-mass spectra with the NIST/NBS, Wiley
library spectra, and literature19 as well as by co-injection with
authentic samples. Medium-pressure liquid chromatography
(MPLC) was performed with a Bu¨chi model 688 apparatus on
columns containing RP-18 Si gel 60 Merck (20-40 µm).
Reversed-phase high-performance liquid chromatography (RP-
HPLC) was performed with a THERMO Finnigan SPECTRA
system.
Plant Material. Ripe fruits of S. devenyense were collected
in October 2003, in the Botanical Garden of the Medical
University of Lublin, Poland. A voucher sample (2080) is kept
in Index Seminum-Anno 2003, Hortus Botanicus Universi-
tatis Mariae Curie-Sklodowska, Lublin, Poland. The plant
material was identified by M. Fransczak-Byc and K. Dab-
rowska.
Extraction and Isolation. The air-dried and powdered
plant material (31 g) was extracted in a Soxhlet apparatus
for 48 h with petroleum ether and then with MeOH. After
evaporation of the solvents, the obtained extracts were 4.3 and
1.6 g, respectively. The MeOH extract was submitted to column
chromatography (3.0 cm) on silica gel 60 Merck (65 g, 40-63
µm) with CH2Cl2/MeOH (from 100:0 to 50:50 gradient) as
eluents to afford 350 fraction of 10 mL each: fractions A1-
A34, CH2Cl2; A35-A46, CH2Cl2/MeOH (99.9:0.1); A47-A61,
CH2Cl2/MeOH (99.8:0.2); A62-A74, CH2Cl2/MeOH (99.6:0.4);
A75-A209, CH2Cl2/MeOH (99.2:0.8); A210-A222, CH2Cl2/
MeOH (99:1); A223-A235, CH2Cl2/MeOH (98.8:1.2); A236-
A248, CH2Cl2/MeOH (98.5:1.5); A249-A262, CH2Cl2/MeOH
(98:2); A263-A332, CH2Cl2/MeOH (95:5); A333-A344, CH2-
Cl2/MeOH (92:8); A345-A352, CH2Cl2/MeOH (85:15); A353-
A360, CH2Cl2/MeOH (50:50).
(+)-Hexanoyllomatin (1): colorless oil; [R]D25 +30.3° (CHCl3,
c 0.1); UV (MeOH) λmax 326 (3.32), 289 (sh) nm; IR (CHCl3)
νmax 2980, 1732, 1606, 1144 cm-1; 1H NMR (CDCl3, 400 MHz)
δ 7.63 (1H, d, J ) 9.5 Hz, H-4), 7.26 (1H, d, J ) 8.6 Hz, H-5),
6.79 (1H, d, J ) 8.6 Hz, H-6), 6.24 (1H, d, J ) 9.5 Hz, H-3),
5.13 (1H, t, J ) 5.0 Hz, H-3′), 3.19 (1H, dd, J ) 15.1, 5.0 Hz,
H-4′a), 2.97 (1H, dd, J ) 15.1, 5.0 Hz, H-4′b), 2.30 (2H, t, J )
7.5 Hz, H-2′′), 1.59 (2H, quint, J ) 7.5 Hz, H-3′′), 1.37 (3H, s,
2′b), 1.34 (3H, s, CH3-2′a), 1.24 (4H, m, H-4′′,5′′), 0.86 (3H, t,
J ) 7 Hz, H-6′′); EIMS m/z 344 (5), 228 (30), 213 (100), 187
(10), 176 (15); HREIMS m/z 344.1630 (cacld for [C20H24O5]+,
344.1624).
25
(+)-Octanoyllomatin (2): amorphous solid; [R]D +30.1°
(CHCl3, c 0.1); UV (MeOH) λmax 326 (3.32), 289 (sh) nm; IR
(CHCl3) νmax 2980, 1731, 1606, 1144 cm-1 1H NMR (CDCl3,
;
400 MHz) δ 7.63 (1H, d, J ) 9.5 Hz, H-4), 7.26 (1H, d, J ) 8.6
Hz, H-5), 6.79 (1H, d, J ) 8.6 Hz, H-6), 6.24 (1H, d, J ) 9.5
Hz, H-3), 5.13 (1H, t, J ) 5.0 Hz, H-3′), 3.19 (1H, dd, J ) 15.1,
5.0 Hz, H-4′a), 2.97 (1H, dd, J ) 15.1, 5.0 Hz, H-4′b), 2.30 (2H,
t, J ) 7.5 Hz, H-2′′), 1.59 (2H, quint, J ) 7.5 Hz, H-3′′), 1.37
(3H, s, CH3-2′b), 1.34 (3H, s, CH3-2′a), 1.24 (8H, m, H-4′′-7′′),
0.86 (3H, t, J ) 7 Hz, H-8′′); 13C NMR (CDCl3) δ 14.0 (C-8′′),
22.6 (C-7′′), 22.9 (C-2′a), 23.0 (C-4′), 24.6 (C-2′b), 24.9 (C-3′′),
28.9 (C-4′′), 29.0 (C-5′′), 31.6 (C-6′′), 34.3 (C-2′′), 69.1 (C-3′),
76.8 (C-2′), 106.9 (C-8), 112.1 (C-4a), 112.5 (C-3), 114.3 (C-6),
126.7 (C-5), 143.8 (C-4), 153.3 (C-8a), 156.2 (C-7), 161.2 (C-2),
173.0 (C-1′′); EIMS m/z 372 (5), 228 (30), 213 (100), 187 (10),
176 (15); HREIMS m/z 372.1931 (calcd for [C22H28O5]+,
372.1937).
25
(+)-Decanoyllomatin (3): amorphous solid; [R]D +29.5°
(CHCl3, c 0.1); UV (MeOH) λmax 326 (3.32), 289 (sh) nm; IR
(CHCl3) νmax 2980, 1730, 1606, 1144 cm-1 1H NMR (CDCl3,
;
400 MHz) δ 7.63 (1H, d, J ) 9.5 Hz, H-4), 7.26 (1H, d, J ) 8.6
Hz, H-5), 6.79 (1H, d, J ) 8.6 Hz, H-6), 6.24 (1H, d, J ) 9.5
Hz, H-3), 5.13 (1H, t, J ) 5.0 Hz, H-3′), 3.19 (1H, dd, J ) 15.1,
5.0 Hz, H-4′a), 2.97 (1H, dd, J ) 15.1, 5.0 Hz, H-4′b), 2.30 (2H,
t, J ) 7.5 Hz, H-2′′), 1.59 (2H, quint, J ) 7.5 Hz, H-3′′), 1.37
(3H, s, CH3-2′b), 1.34 (3H, s, CH3-2′a), 1.24 (12H, m, H-4′′-9′′),
0.86 (3H, t, J ) 7 Hz, H-10′′); 13C NMR (CDCl3) δ 14.1 (C-
10′′), 22.6 (C-9′′), 22.9 (C-2′b), 23.0 (C-4′), 24.6 (C-2′a), 24.9
(C-3′′), 29.0 (C-4′′), 29.2, 29.4 (C-5′′,6′′,7′′), 31.8 (C-8′′), 34.3 (C-
2′′), 69.1 (C-3′), 76.8 (C-2′), 106.9 (C-8), 112.1 (C-4a), 112.5 (C-
3), 114.3 (C-6), 126.7 (C-5), 143.9 (C-4), 153.3 (C-8a), 156.2 (C-
7), 161.2 (C-2), 173.1 (C-1′′); EIMS m/z 400 (5), 228 (30), 213
(100), 187 (10), 176 (15); HREIMS m/z 400.2255 (calcd for
[C24H32O5]+, 400.2250).
(+)-Dodecanoyllomatin (4): amorphous solid; [R]D25 +29.3°
(CHCl3, c 0.1); UV (MeOH) λmax 326 (3.32), 289 (sh) nm; IR
(CHCl3) νmax 2980, 1730, 1606, 1144 cm-1 1H NMR (CDCl3,
;
400 MHz) δ 7.63 (1H, d, J ) 9.5 Hz, H-4), 7.26 (1H, d, J ) 8.6
Hz, H-5), 6.79 (1H, d, J ) 8.6 Hz, H-6), 6.24 (1H, d, J ) 9.5
Hz, H-3), 5.13 (1H, t, J ) 5.0 Hz, H-3′), 3.19 (1H, dd, J ) 15.1,
5.0 Hz, H-4′a), 2.97 (1H, dd, J ) 15.1, 5.0 Hz, H-4′b), 2.30 (2H,
t, J ) 7.5 Hz, H-2′′), 1.59 (2H, quint, J ) 7.5 Hz, H-3′′), 1.37
(3H, s, CH3-2′b), 1.34 (3H, s, CH3-2′a), 1.24 (16H, m, H-4′′-
11′′), 0.86 (3H, t, J ) 7 Hz, H-12′′); 13C NMR (CDCl3) δ 14.1
(C-12′′), 22.6 (C-11′′), 22.9 (C-2′a), 23.0 (C-4′), 24.6 (C-2′b), 24.9
(C-3′′), 29.0 (C-4′′), 29.2-29.4 (C-5′′-9′′), 31.8 (C-10′′), 34.3 (C-
2′′), 69.1 (C-3′), 76.7 (C-2′), 106.9 (C-8), 112.1 (C-4a), 112.5 (C-
3), 114.3 (C-6), 126.7 (C-5), 143.8 (C-4), 153.3 (C-8a), 156.2 (C-
7), 161.2 (C-2), 173.1 (C-1′′); EIMS m/z 428 (5), 228 (30), 213
(100), 187 (10), 176 (15); HREIMS m/z 428.2558 (cacld for
[C26H36O5]+, 428.2562).
Fractions A35-A133 (38 mg) were rechromatographed in
HPLC [Nucleosil C18, 100-7 (7 µm, 250 × 21 mm), H2O/MeOH
(60:40 to 40:60 in 60 min), flow rate 1 mL/min] to afford four
lomatin esters: 1 (2 mg), 2 (13 mg), 3 (15 mg), and 4 (5 mg).
Fractions A134-A144 (96 mg) were rechromatographed by
column chromatography (1.3 cm, 40-63 µm) on Merck silica
gel 60 (4.0 g, 40-63 µm) with CH2Cl2/MeOH (from 100:0 to
99.5:0.5 gradient) as eluents to afford 60 fractions of 10 mL
each: Fractions B10-B15 afforded (+)-4′-decanoyl-cis-khel-
lactone (5) (15 mg), B17-B22 (+)-3′-decanoyl-cis-khellactone
(6) (10 mg), B25-B28 laserpitin (2 mg), and B32-B42 3′-
angeloyl-cis-khellactone (3 mg).
Fractions A176-A185 afforded (+)-cis-khellactone (3.6 mg).
Fractions A276-A284 afforded devenyol (7) (7 mg).
Fractions A353-A360 (400 mg) were rechromatographed by
MPLC on RP-18 silica gel 60 Merck (20-40 µm) with H2O/
MeOH (from 100:0 to 70:30 gradient) as eluents to afford 130
(+)-4′-Decanoyl-cis-khellactone (5): amorphous solid;
[R]D25 +47.2° (CHCl3, c 0.1); UV (MeOH) λmax (log ꢀ) 327 (3.17),
288 (sh) nm; IR (MeOH, CaF2) νmax 1725, 1607 cm-1; 1H NMR
(CDCl3 400 MHz) δ 7.60 (1H, d, J ) 9.5 Hz, H-4), 7.35 (1H, d,
J ) 8.6 Hz, H-5), 6.79 (1H, d, J ) 8.6 Hz, H-6), 6.41 (1H, d, J
) 4.9 Hz, H-4′), 6.23 (1H, d, J ) 9.6 Hz, H-3), 4.02 (1H, d, J

1640 Journal of Natural Products, 2005, Vol. 68, No. 11
Widelski et al.
) 4.9 Hz, H-3′), 2.43 (2H, m, H-2′′), 1.67 (2H, m, H-3′′), 1.46
(3H, s, CH3-2′b), 1.43 (3H, s, CH3-2′a), 1.25 (12H, br s, H-4′′-
H-9′′), 0.86 (3H, t, J ) 7 Hz, H-10′′); 13C NMR (CDCl3) δ 14.1
(C-10′′), 21.2 (C-2′b), 22.6 (C-9′′), 24.8 (C-3′′), 25.4 (C-2′a), 29.1
(C-4′′), 29.2, 29.4 (C-5′′,6′′,7′′), 31.8 (C-8′′), 34.3 (C-2′′), 63.6 (C-
4′), 71.3 (C-3′), 78.6 (C-2′), 106.9 (C-8), 112.3 (C-4a), 113.0 (C-
3), 114.5 (C-6), 129.3 (C-5), 143.4 (C-4), 154.1 (C-8a), 156.9 (C-
7), 160.0 (C-2), 175.1 (C-1′′); HREIMS m/z 416.2255 (cacld for
[C24H32O6]+, 416.2250).
(+)-3′-Decanoyl-cis-khellactone (6): amorphous solid;
[R]D25 +45.2° (CHCl3, c 0.1); UV (MeOH) λmax (logꢀ) 327 (3.17),
288 (sh) nm; IR (MeOH, CaF2) νmax 1725, 1607 cm-1; 1H NMR
(CDCl3 400 MHz) δ 7.64 (1H, d, J ) 9.5 Hz, H-4), 7.33 (1H, d,
J ) 8.6 Hz, H-5), 6.79 (1H, d, J ) 8.6 Hz, H-6), 6.25 (1H, d, J
) 9.6 Hz, H-3), 5.41 (1H, d, J ) 4.9 Hz, H-4′), 5.17 (1H, d, J
) 4.9 Hz, H-3′), 2.35 (2H, m, H-2′′), 1.67 (2H, m, H-3′′), 1.47
(3H, s, H-CH3-2′a), 1.39 (3H, s, CH3-2′b), 1.25 (12H, br s, H-4′′-
H-9′′), 0.86 (3H, t, J ) 7 Hz, H-10′′); 13C NMR (CDCl3) δ 14.1
(C-10′′), 22.5 (C-2′a), 22.6 (C-9′′), 24.9 (C-3′′), 25.4 (C-2′b), 29.1
(C-4′′), 29.2, 29.4 (C-5′′,6′′,7′′), 31.6 (C-8′′), 34.2 (C-2′′), 60.0 (C-
4′), 72.1 (C-3′), 77.7 (C-2′), 110.7 (C-8), 112.3 (C-4a), 112.6 (C-
3), 114.6 (C-6), 128.8 (C-5), 144.0 (C-4), 154.1 (C-8a), 156.0 (C-
7), 160.7 (C-2), 173.2 (C-1′′); HREIMS m/z 416.2258 (cacld for
[C24H32O6]+, 416.2250).
amorphous solid; [R]25 -169.5° (MeOH, c 0.4); UV (MeOH)
D
λmax (log ꢀ) 312 (3.85), 256 (sh) nm, no shift with NaOAc; IR
(MeOH, CaF2) νmax 1710, 1606, 1250 cm-1; 1H NMR (CD3OD)
δ 1.302 (3H, s, H-4′), 1.305 (3H, s, H-5′), 3.12 (2H, m, H-1′a,
1′b), 3.39 (1H, t, J ) 9.0 Hz, H-4′′), 3.47 (1H, t, J ) 9.0 Hz,
H-3′′), 3.49 (1H, m, H-5′′), 3.56 (1H, t, J ) 9.0, 7.6 Hz, H-2′′),
3.63 (1H, dd, J ) 9.4, 4.1 Hz, H-2′), 3.71 (1H, dd, J ) 12.0, 5.8
Hz, H-6b′′), 3.93 (1H, dd, J ) 12.0, 2.2 Hz, H-6a′′), 4.96 (1H,
d, J ) 7.6 Hz, H-1′′), 6.29 (1H, d, J ) 9.5 Hz, H-3), 7.26 (1H,
d, J ) 8.6 Hz, H-6), 7.49 (1H, d, J ) 8.6 Hz, H-5), 7.91 (1H, d,
J ) 9.5 Hz, H-4); 13C NMR (CD3OD) δ 25.1 (C-4′), 25.8 (C-1′),
25.9 (C-5′), 62.5 (C-6′′), 71.2 (C-4′′), 75.1 (C-2′′), 77.9 (C-5′′),
78.5 (C-3′′), 79.2 (C-2′), 74.0 (C-3′), 103.2 (C-1′′), 113.3 (C-6),
114.0 (C-3), 115.4 (C-4a), 118.2 (C-8), 128.4 (C-5), 146.2 (C-4),
154.4 (C-8a), 161.0 (C-7), 163.2 (C-2); HRFABMS m/z 427.1612
(cacld for [C20H26O10 + H]+, 427.1604).
Devenyoside C: (-)-3′-O-â-D-Glucopyranosyl 8-[(2S),3-
dihydroxy-3-methylbutyl]-7-hydroxychromen-2-one 7-O-
â-D-glucopyranoside (10): amorphous solid; [R]25 -36°
D
(MeOH, c 0.4); UV (MeOH) λmax (log ꢀ) 313 (3.67), 256 (sh) nm,
no shift with NaOAc; IR (MeOH, CaF2) νmax 1710,1607, 1250
cm-1; 1H NMR (CD3OD) δ 1.40 (3H, s, H-4′), 1.43 (3H, s, H-5′),
3.12 (2H, m, H-1′a,b), 3.21 (1H, dd, J ) 9.0, 7.7 Hz, H-2′′),
3.29 (1H, m, H-5′′), 3.32 (1H, t, J ) 9.0 Hz, H-4′′), 3.39 (2H, t,
J ) 9.0 Hz, H-3′′, 4′′′), 3.48 (2H, t, J ) 8.9 Hz, H-3′′′), 3.49
(1H, m, H-5′′′), 3.56 (1H, dd, J ) 8.9, 7.6 Hz, H-2′′′), 3.65 (1H,
dd, J ) 12.2, 5.4 Hz, H-6b′′), 3.71 (1H, dd, J ) 12.2, 5.8 Hz,
H-6b′′′), 3.79 (1H, dd, J ) 12.0, 2.0 Hz, H-6a′′), 3.81 (1H, dd,
J ) 9.0, 3.8 Hz, H-2′), 3.94 (1H, dd, J ) 12.2, 2.2 Hz, H-6a′′′),
4.59 (1H, d, J ) 7.7 Hz, H-1′′), 4.98 (1H, d, J ) 7.6 Hz, H-1′′′),
6.29 (1H, d, J ) 9.6 Hz, H-3), 7.24 (1H, d, J ) 8.7 Hz, H-6),
7.49 (1H, d, J ) 8.7 Hz, H-5), 7.91 (1H, d, J ) 9.6 Hz, H-4);
13C NMR (CD3OD) δ 21.8 (C-4′), 23.6 (C-5′), 25.2 (C-1′), 62.5
(C-6′′, 6′′′), 71.2 (C-4′′, 4′′′), 74.8 (C-2′′′), 75.0 (C-2′′), 77.0 (C-
5′′′), 77.5 (C-5′′), 77.9 (C-3′′), 78.0 (C-3′′′), 78.2 (C-2′), 81.6 (C-
3′), 98.2 (C-1′′), 102.8 (C-1′′′), 112.8 (C-6), 113.7 (C-3), 115.4
(C-4a), 118.1 (C-8), 128.1 (C-5), 146.0 (C-4), 154.3 (C-8a), 161.2
(C-7), 163.2 (C-2); HRFABMS m/z 589.2139 (cacld for [C26H36O15
+ H]+, 589.2132).
Devenyol: (-)-8-[(2S),3-Dihydroxy-3-methylbutyl]-7-
hydroxychromen-2-one (7): amorphous solid; [R]D25 -44.6°
(MeOH, c 0.1); UV (MeOH) λmax (log ꢀ) 326 (3.55), 256 (sh) nm;
UV (MeOH+NaOAc) λmax (log ꢀ) 371 (3.35) nm; IR (MeOH,
CaF2) νmax 1711, 1607, 1250 cm-1 1H NMR (CD3OD) δ 1.29
;
(3H, s, H-4′), 1.30 (3H, s, H-5′), 2.93 (1H, dd, J ) 13.8, 10.1
Hz, H-1′a), 3.17 (1H, dd, J ) 13.8, 1.5 Hz, H-1′b), 3.68 (1H,
dd, J ) 10.1, 1.5 Hz, H-2′), 6.19 (1H, d, J ) 9.3 Hz, H-3), 6.84
(1H, d, J ) 8.5 Hz, H-6), 7.35 (1H, d, J ) 8.5 Hz, H-5), 7.86
(1H, d, J ) 9.3 Hz, H-4); 13C NMR (CD3OD) δ 25.4 (C-4′), 25.6
(C-5′), 26.4 (C-1′), 74.0 (C-3′), 79.5 (C-2’), 111.9 (C-3), 113.4
(C-4a), 115.5 (C-8), 114.2 (C-6), 128.3 (C-5), 146.7 (C-4), 155.1
(C-8a), 163.8 (C-2), 161.3 (C-7); CI-MS (CH4) m/z 265 (M +
H)+; HRFABMS m/z 265.1069 (cacld for [C14H16O5 + H]+,
265.1076).
Devenyoside A: 2′-epi-Tortuoside: (-)-3′-O-â-D-gluco-
pyranoside of 8-[(2S),3-dihydroxy-3-methylbutyl]-7-hy-
Alkaline Hydrolysis of Lomatin Esters (3, 4) and 3′-
Decanoyl-cis-khellactone (6). To a solution of 3 (10 mg) or
4 (5 mg) or 6 (10 mg) in MeOH (2 mL) was added methanolic
KOH (2 mL, 1 N), and the reaction mixture was refluxed for
1 h. The reaction mixture was diluted with H2O, concentrated
to remove organic solvent, acidified with H2SO4, and extracted
with EtOAc. The EtOAc phase was washed with NaHCO3 and
submitted to preparative TLC (cyclohexane/EtOAc) to give
either (+)-lomatin in the case of 3 and 4 or (+)-cis-khellactone
in the case of 6. The aqueous phase was acidified with H2SO4,
extracted with Et2O, and submitted to GC-MS analysis (for
the identification of decanoic and dodecanoic acid).
Acid Hydrolysis of 4′-Decanoyl-cis-khellactone (5). To
a solution of 5 (10 mg) in MeOH (3 mL) was added methanolic
HCl (3 mL, 1 N), and the reaction mixture was refluxed for
24 h. The mixture was diluted with H2O, concentrated to
remove organic solvent, alkalized with NaHCO3, and extracted
with EtOAc. The EtOAc phase was washed with H2O and
subjected to preparative TLC (cyclohexane/EtOAc) to give (+)-
cis-methylkhellactone and (-)-trans-methylkhellactone, identi-
fied by 1H NMR and specific rotation, identical with previously
reported data.16 The aqueous NaHCO3 phase was acidified
with H2SO4, extracted with Et2O, and subjected to GC-MS
analysis (for the identification of decanoic acid).
25
droxychromen-2-one (8): amorphous solid; [R]D -38.8°
(MeOH, c 0.4); UV (MeOH) λmax (log ꢀ) 326 (3.63), 256 (3.26)
nm; UV (MeOH+NaOAc) λmax (log ꢀ) 374 (4.09) nm; IR (MeOH,
1
CaF2) νmax 1709, 1606, 1583, 1211 cm-1; H NMR (CD3OD) δ
1.36 (3H, s, H-4′), 1.40 (3H, s, H-5′), 2.96 (1H, dd, J ) 13.8,
10.1 Hz, H-1′a), 3.13 (1H, dd, J ) 13.8, 1.5 Hz, H-1′b), 3.23
(1H, dd, J ) 8.5, 7.7 Hz, H-2′′), 3.29 (1H, m, H-5′′), 3.32 (1H,
t, J ) 8.5 Hz, H-4′′), 3.41 (1H, t, J ) 8.5 Hz, H-3′′), 3.62 (1H,
dd, J ) 12.0, 5.2 Hz, H-6b′′), 3.79 (1H, dd, J ) 10.1, 1.5 Hz,
H-2′), 3.80 (1H, dd, J ) 12.0, 2.0 Hz, H-6a′′), 4.58 (1H, d, J )
7.7 Hz, H-1′′), 6.02 (1H, d, J ) 9.3 Hz, H-3), 6.72 (1H, d, J )
8.5 Hz, H-6), 7.23 (1H, d, J ) 8.5 Hz, H-5), 7.76 (1H, d, J )
9.3 Hz, H-4); 13C NMR (CD3OD) δ 21.9 (C-4′), 23.9 (C-5′), 26.3
(C-1′), 62.5 (C-6′′), 71.5 (C-4′′), 75.1 (C-2′′), 77.5 (C-5′′), 77.9
(C-3′′), 79.6 (C-2′), 81.9 (C-3′), 98.6 (C-1′′), 109.0 (C-3), 111.6
(C-4a), 115.6 (C-8), 116.3 (C-6), 128.3 (C-5), 147.1 (C-4), 155.5
(C-8a), 164.9 (C-2), 167.3 (C-7); HRFABMS m/z 427.1610 (cacld
for [C20H26O10 + H]+, 427.1604).
Acetylation of Devenyoside A (2′-epi-Tortuoside) (8a).
A solution of 8 (5 mg) and Ac2O (1 mL) in dry pyridine (1 mL)
was stirred for 12 h at room temperature. Evaporation of the
reagents under reduced pressure afforded the hexaacetyl
derivative 8a: 1H NMR (CDCl3) δ 1.33 (3H, s, H-4′), 1.37 (3H,
s, H-5′), 1.83 (3H, s, 2′-OAc), 2.00 (3H, s, OAc), 2.01 (3H, s,
OAc), 2.03 (3H, s, OAc), 2.07 (3H, s, OAc), 2.42 (3H, s, 7-OAc),
3.04 (2H, m, H-1′), 3.73 (1H, m, H-5′′), 4.11 (1H, dd, J ) 12.0,
2.4 Hz, H-6b′′), 4.22 (1H, dd, J ) 12.0, 5.8 Hz, H-6a′′), 4.81
(1H, d, J ) 7.9 Hz, H-1′′), 4.98 (1H, t, J ) 9.6 Hz, H-2′′), 5.03
(1H, t, J ) 9.6 Hz, H-4′′), 5.21 (1H, dd, J ) 7.5, 5.5 Hz, H-2′),
5.26 (1H, t, J ) 9.6 Hz, H-3′′), 6.38 (1H, d, J ) 9.6 Hz, H-3),
7.01 (1H, d, J ) 8.4 Hz, H-6), 7.36 (1H, d, J ) 8.4 Hz, H-5),
7.66 (1H, d, J ) 9.6 Hz, H-4).
Enzymatic Hydrolysis of Devenyosides A-C (8-10).
To a solution of 8, 9, or 10 (10 mg) in H2O (2 mL) was added
â-D-glucosidase (15 mg), and the reaction mixture was stirred
for 48 h at 37 °C. The mixture was then extracted with EtOAc,
and the organic phase was evaporated to dryness. Purification
of the residue with pTLC afforded in all cases devenyol (7),
1
identified by H NMR and specific rotation.
References and Notes
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DevenyosideB: (-)-7-O-â-D-Glucopyranosideof8-[(2S),3-
dihydroxy-3-methylbutyl]-7-hydroxychromen-2-one (9):

Coumarins from the Fruits of Seseli devenyense
Journal of Natural Products, 2005, Vol. 68, No. 11 1641
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