Phytochemical investigation of the dichloromethane extract of the dried roots of Ferula badrakema resulted in the identification of one new and six known compounds. Known compounds were sesquiterpene coumarins: mogoltacin, feselol, badrakemin acetate, ferocaulidin, conferone and conferol acetate. The new compound was a sesquiterpene, named badrakemonin. The structures of these compounds were elucidated by extensive NMR spectroscopic methods including 1D-(1H and 13C) and 2D-NMR (HSQC, HMBC, and ROESY) as well as MS experiments.
aDepartment of Pharmacognosy, Biotechnology
Research Center, School of Pharmacy, Mashhad University of Medical Sciences,
Mashhad, Iran. bDepartment of Pharmacognosy, School of Pharmacy, Mashhad
University of Medical Sciences, Mashhad, Iran. cDipartimento di Scienze
Farmaceutiche, Universit? degli Studi di Salerno, Via Ponte Don Melillo, 84084,
Phytochemical investigation of the dichloromethane extract of the dried roots of
Ferula badrakema resulted in the identification of one new and six known
compounds. Known compounds were sesquiterpene coumarins: mogoltacin, feselol,
badrakemin acetate, ferocaulidin, conferone and conferol acetate. The new
compound was a sesquiterpene, named badrakemonin. The structures of these
compounds were elucidated by extensive NMR spectroscopic methods including
1D-(1H and 13C) and 2D-NMR (HSQC, HMBC, and ROESY) as well as MS experiments.
Genus of Ferula which belongs to tribe Peucedaneae, subfamily of Apioideae,
family of Umbelliferae with 133 species distributed throughout Mediteranean area
and central Asia, especially in the former USSR and neighboring countries such
as Iran (1-3). More than 70 species of Ferula have already been investigated
chemically (4-6). Several species of this genus have been used in folk medicine
for their antispasmodic, carminative, digestive, expectorant, sedative, antihysteric, laxative, aphrodisiac, antiseptic, and analgesic activities (7).
The Iranian flora comprises 30 species of Ferula, of which some are endemic (2,
8). The popular Persian name of the most of these species is ?Koma? (8).
Chemistry of this genus has been studied by many investigators (9) and is well
documented as a good source of biologically active compounds such as
sesquiterpene derivatives (10-16) and sulfur containing compounds (17-22). A few
sesquiterpene coumarin glycosides have also been reported from Ferula species
(6, 23). Sesquiterpene derivatives, especially sesquiterpene coumarins, were
stored in the roots of the plants; therefore the roots are better source for
isolating these compounds than the aerial parts. However, to the best of our
knowledge, there is no report about eremophilane-type sesquiterpenes from this
Ferula badrakema Kos.-Pol. (Syn = Ferulaafghanistanica) is a resinous plant
with a strong odor which is endemic to Iran (24). The roots of this plant have
been used in folk medicine as antiepileptic and antispasmodic drug (7). Similar
to other species of the genus Ferula, F. badrakema is a rich source of
sesquiterpene coumarins (25). Previously, some sesquiterpene coumarins have been
isolated from the roots of the plant (25, 26). In the present study, we have
reported the phytochemical study of the plant F. badrakema.
The roots of F. badrakema were collected from the Tandoureh park, Khorasan
Razavi, Iran, in July 2006. The plant was identified by the department of
pharmacognosy, School of pharmacy, Mashhad University of Medical Science. A
voucher specimen (No. 1002) has been deposited at the Herbarium of the School of
General experimental procedures
Melting points were determined on a Electrothermal 9100 apparatus and are
uncorrected. The optical rotation was measured on a Polax-2L ATAGO Polarimeter.
UV spectra were obtained using a Shimadzu PC-1650 spectrophotometer. ESIMS
analyses were performed using a ThermoFinnigan LCQ Deca XP Max ion-trap mass
spectrometer equipped with Xcalibur software.
NMR spectra were measured on a Bruker DRX 500 (Bruker Biospin, Rheinstetten,
Germany). 1H NMR, 13C NMR, DEPT, 1H-1H COSY, HMBC, HSQC, and ROESY spectra were
measured using an inverse-detection probe (5 mm). The operating frequencies were
500.13 MHz for acquiring 1H NMR and 125.75 MHz for 13C NMR spectra. Samples were
measured at 300 K in CDCl3 with TMS as the internal standard. Column
chromatography was conducted with silica gel 230-400 mesh (Merck). Preparative
TLC was performed on GF254s plates (20 ? 20 cm, Merck) and observation of plates
was carried out under UV CAMAG spectrometer (254 nm).
Extraction and isolation
Dried, powdered roots of F. badrakema (383.54 g) were extracted with
dichloromethane by maceration. The combined dichloromethane extracts were
concentrated in vacuo to give a red extract (21.17 g). Part of the extract
(13.37 g) was subjected to column chromatography on silica gel (5 cm?50 cm)
using gradient compositions of ethyl acetate (EtOAc)-Petroleum ether as eluent [EtOAc-Petroleum
ether (1:10, 2200 mL), (1:9, 2000 mL), (1:8, 1800 mL), (1:7, 2400 mL), (1:6,
2100 mL), (1:5, 1200 mL), (1:4, 2000 mL), (1:3, 800 mL), (1:2, 900 mL), (1:1,
2000 mL) and EtOAc (3000 mL)]. The fractions were compared by TLC (on silica gel
using EtOAc-petroleum ether as eluent ), and those giving similar spots were
combined. Twenty-three fractions were finally obtained. Fraction 1 was subjected
to silica gel PTLC (EtOAc-petroleum ether, 1:10) to give badrakemonin (4.4 mg).
Fraction 10 was also developed on silica gel PTLC (acetone-petroleum ether,
1:4.5) to yield conferol acetate (1.7 mg). Fractions 11 and 12 were subjected to
silica gel PTLC (EtOAc-petroleum ether, 2:3) to give badrakemin acetate (14.3
mg). Fractions 13, 14 and 15 also needed more purification with silica gel PTLC
(EtOAc-petroleum ether, 2:3) to afford conferone (13.5 mg), mogoltacin (38.8 mg)
and feselol (49.2 mg), respectively. Fraction 17 was further purified by silica
gel PTLC (EtOAc- petroleum ether, 3:4) to yield mogoltacin (71.2 mg). Fractions
18 and 19 were also further purified by silica gel PTLC (EtOAc-petroleum ether,
3:2) to give ferocaulidin (64.9 mg).
Results and Discussion
Normal-phase column chromatography of the dichloromethane extract of roots,
followed by preparative TLC, afforded a new sesquiterpene, badrakemonin, and six
known sesquiterpene coumarins, namely mogoltacin, feselol, badrakemin acetate,
ferocaulidin, conferone and conferol acetate (Figure 1). The structures of the
mentioned known compounds were confirmed according to the melting points, NMR
experiments and literature (6, 9, 27).
The 1H and 13C NMR resonances (Table 1) of badrakemonin were assigned by
different 2D NMR experiments. The 1H NMR spectrum showed resonances
characteristic for three methyl singlets at δΗ 1.50 (H-10), 1.83 (H-8) and 1.86
(H-9), and two methine resonances at δΗ 1.84 (H-15', 1H, septet,
J = 6.6 Hz) and
5.76 (H-11, 1H, dd, J = 10.8 and 17.4 Hz).
The 13C NMR resonances showed 15 carbon signals, which could be resolved by DEPT
and HSQC experiments into five methyls at δC 16.8 (C-14), 17.1 (C-15), 23.8
(C-8), 23.9 (C-10) and 24.9 (C-9), one tertiary alcoholic carbon at δC 80.0
characteristic for C-6, three methylenes at δC 31.4 (C-5), 37.0 (C-4) and 111.2
(C-12), two methines at δC 36.0 (C-13) and 146.0 (C-11), and five quaternary
carbons at δC 44.9 (C-3), 80.0 (C-6), 137.3 (C-2), 145.6 (C-7) and one carbonyl
function which was indicated by the downfield signal at δC 211.9 (C-1). In the
HMBC spectrum, the correlations of H-4 (δH 1.34 and 1.53) with C-3 (δC 44.9);
H-5 (δH 1.75 and 2.14) with C-6 (δC 80.0); H-8 (δH 1.83) with C-2 (δC 137.3) and
C-7 (δC 145.6); H-9 (δH 1.86) with C-2 (δC 137.3) and C-7 (δC 145.6); H-10 (δH
1.50) with C-3 (δC 44.9); H-11 (δH 5.76) with C-3 (δC 44.9) and C-12 (δC 111.2);
H-12 (δH 4.96 and 4.98) with C-11 (δC 146.0); H-13 (δH 1.84) with C-6 (δC 80.0);
H-14 (δH 0.98) with C-6 (δC 80.0) and C-13 (δC 36.0); and H-15 (δH 0.80) with
C-6 (δC 80.0) and C-13 (δC 36.0) were observed. The proposed structure was
further supported by 1H-1H COSY data.
The stereochemistry of the chemical groups at C-3 and C-6 in 1 was determined on
the basis of the ROESY experiment, in which cross-peaks were observed from
H-10/H-4β pairs and H-10/H-5β pairs (Figure 2).
This research was supported by a grant from Mashhad University of Medical
Sciences Research Council.
(1) Evans WC. Trease and Evans? Pharmacognosy.
13th ed. Bailliere Tindall, London (1989) 205-206.
Mozaffarian V. The Family of Umbelliferae in Iran- Keys and Distribution.
Research Institute of Forests and Rangelands Press, Tehran (1983) 114-116.
Heywood VH. Flowering Plants of the World. Croom Helm, London (1985) 219-221.
Diab Y, Dolmazon R and Bessiere JM. Daucane aryl esters composition from the
Lebanese Ferulahermonis Boiss. (zallooh root). Flav. Fragr. J. (2001) 16:
Iranshahi M, Amin GR and Shafiee A. A new coumarin from Ferulapersica.
Biol. (2004) 42: 440-442.
Abd El-Razek MH, Ohta S and Hirata T. Terpenoid coumarins of the genus Ferulaheterocycles.(2003) 60: 689-716.
Zargari A. Medicinal Plants. vol. 2. Tehran University Publications, Tehran
Mozaffarian V. A Dictionary of Iranian Plant Names. Farhang-e Moaser, Tehran
Murray RDH, Mendez J and Brown SA. The Natural Coumarins. John Wiley & Sons
Inc., New York (1982) 555-559.
Iranshahi M, Shahverdi AR, Mirjani R, Amin GR and Shafiee A. Umbelliprenin from
Ferulapersica roots inhibits the red pigment production in Serratiamarcescens.
Z. Naturforschung. (2004) 59c: 506-508.
Kogure K, Yamauchi I, Tokumura A, Kondou K, Tanaka N, Takaishi Y and Fukuzawa K.
Novel antioxidants isolated from plants of the genera Ferula, Inula,
Rheum collected in Uzbekistan. Phytomedicine (2004) 11: 645-651.
Tamemoto K, Takais Y, Chen B, Kawazoe K, Shibata H, Higuti T, Honda G, Ito M,
Takeda Y, Kodzhimatov OK and Ashurmetov O. Sesquiterpenoids from the fruits of
Ferulakuhistanica and antibacterial activity of the constituents of
F. kuhistanica. Phytochem. (2001) 58: 763-767.
Iranshahi M, Kalategi F, Rezaiee R, Shahverdi AR, Ito C, Furukawa H, Tokuda H
and Itoigawa M. Cancer chemopreventive activity of terpenoid coumarins from
Ferula species. Planta Med. (2008) 74: 147-150.
Iranshahi M, Arfaa P, Ramezani M, Jaafari MR, Sadeghian H, Bassarello C,
Piacente S and Pizza C. Sesquiterpene coumarins from Ferulaszowitsiana and
vitro antileishmanial activity of 7-prenyloxycoumarins against promastigotes.
Phytochem. (2007) 68: 554-561.
Shahverdi AR, Saadat F, Khorramizadeh MR, Iranshahi M and Khoshayand MR. Two
matrix metalloproteinase from Ferulapersica var. persica.
Motai T, Daikonya A and Kitanaka S. Sesquiterpene coumarins from Ferulafukanensis and nitric oxide production inhibitory effects. J. Nat. Prod. (2004)
Al-said MS, Abdel Sattar E, El-Feraly F, Nahrstedt A and Coen M. New sulphides
from Ferularutabensis. Int. J. Pharmacog. (1996) 43: 189-193.
Iranshahi M, Amin G, Amini M and Shafiee A. Sulfur containing derivatives from
Ferulapersica var. latisecta. Phytochem. (2003) 63: 965-966.
Iranshahi M, Amin G, Salehi-Sourmaghi MH, Shafiee A and Hadjiakhoondi A.
Sulphur-containing compounds in the essential oil of the root of Ferulapersica
Willd. var. persica. Flav. Fragr. J. (2006) 21: 260-261.
Rajanikanth B, Ravindranath B and Shankaranaryana ML. Volatile polysulphides of
asafoetida. Phytochem. (1984) 23: 899-900.
Takeoka G. Volatile constituents of asafetida. In: Aroma Active Compounds in
Foods. American Chemical Society, Washington (year) Pages.
Zhi-da M, Qi-fi M, Mizuno M, Tanaka T and Iinuma M. Polysulfanes in the volatile
oils of Ferula species. Planta Med. (1987) 53: 300-302.
Iranshahi M, Mojarab M, Sadeghian H, Hanafi-Bojd MY and Schneider B. Polar
secondary metabolites of Ferulapersica roots. Phytochem. (2008) 69: 473-478.
Rechinger KH, Lemond JM and Hedge IC. FloraIranica (Umbelliferae). No. 162,
Akademischev Druk-u. Verlagsanstalt, Graz (1994) 411.
Bukreeva TV and Pimenov MG. Coumarins from the roots of Ferula badrakema.
Prir. Soedin. (1991) 27: 718.
Kir?yalov NP. The coumarin, badrakemin from the roots of Ferula badrakema.
Prir. Soedin. (1967) 3: 363.
Lee E and Mabry TJ. Sesquiterpene coumarin ethers of Ferulatingitana.
Prod. (1985) 48: 326-327.