Cecilia Noccioli

Pterocarpans from Bituminaria morisiana and Bituminaria bituminosa.

The aerial parts of Mediterranean papilionaceous plants Bituminaria morisiana and B. bituminosa afforded, along with known phenolics, the prenylated pterocarpans bitucarpin A and B, whose structure was elucidated by spectroscopic techniques. A known isoflavonoid (8-prenyldaidzein) was also obtained for the first time as a genuine plant constituent. The accumulation of pterocarpans at the expense of biogenetically more primitive shikimate metabolites like furanocoumarins or isoflavonoids supports the inclusion of this plant, once part of the genus Psoralea, into the distinct genus Bituminaria.

Estradiol-antagonistic Activity of Phenolic Compounds from Leguminous Plants

Natural flavonoids are currently receiving much attention because of their estrogenic and antiestrogenic properties. Six isoflavones (isoprunetin, isoprunetin 7‐O‐β‐d‐glucopyranoside, isoprunetin 4′,7‐di‐O‐β‐d‐glucopyranoside, genistein, genistein 7‐O‐β‐d‐glucopyranoside, daidzein), four flavones (luteolin, luteolin 7‐O‐β‐d‐glucopyranoside, luteolin 4′‐O‐β‐d‐glucopyranoside, licoflavone C), isolated from Genista morisii and G. ephedroides (two Leguminosae plants of the Mediterranean area) together with two structurally related pterocarpans, bitucarpin A and erybraedyn C, isolated from Bituminaria bituminosa (Leguminosae), were tested for the antagonist activity by a yeast based estrogen receptor assay (Saccharomyces cerevisiae RMY326 ER‐ERE). Most compounds inhibited the estradiol‐induced transcriptional activity in a concentration dependent manner. In particular, for the flavone luteolin 77% inhibition of the induced β‐galactosidase activity was observed. Interestingly, licoflavone C exhibited a dose‐dependent antagonistic activity at concentrations up to 10−4 m, but stimulated β‐galactosidase expression at higher concentrations resulting in a U‐shaped‐like dose‐response curve. Copyright

Lupane-triterpenes from Bupleurum flavum

Two new triterpenoids, lup-20(29)-en-3α,28-diol (1) and lup-20(29)-en-3α,28,30-triol (2), have been isolated from aerial parts of Bupleurum flavum, a native plant of the Eastern Mediterranean area (Bulgaria, Greece, and Turkey). Their structures were elucidated by spectral analysis. The known compounds betulin, betulinic acid, jasminol, together with the lignan nemerosin and eight flavonoids (kaempferol, isokaempferide, gossipetin, quercetin, luteolin, isorhamnetin 3-O-β-D-glucopyranoside, isorhamnetin 3-rutinoside, and rutin) were also characterized.

Chemical composition and volatile constituents of Anthyllis barba-jovis

Six flavonol glycosides were isolated and identified from the aerial parts of Anthyllis barba-jovis L. (Leguminosae), together with two coumarins and D-pinitol, a taxonomic marker of Leguminosae family. The structural elucidation of all compounds was based on their 1H- and 13C-NMR spectral data and bidimensional experiments. The total flavonoid content was also determined, according to the method described in the Italian pharmacopoeia. In order to complete the phytochemical investigation on A. barba-jovis, the essential oils from flowers and seeds were obtained by hydrodistillation and analysed by GC-MS; fresh flowers were analysed also directly by solid phase micro extraction (SPME). §Dedicated to the memory of Prof. I. Morelli (1940–2005).

Chemical Composition of Essential Oils and Aromatic Waters from Different Italian Anthemis maritima Populations

The chemical composition of the essential oils and aromatic waters isolated from six Italian Anthemis maritima populations was determined by GC‐FID and GC/MS analyses. In total, 122 and 100 chemical compounds were identified in the essential oils and the aromatic waters, respectively. The main compound classes represented in the oils were monoterpene hydrocarbons, oxygenated monoterpenes, sesquiterpene hydrocarbons, oxygenated sesquiterpenes, and terpene esters. Multivariate chemometric techniques such as cluster analysis (CA) and principal coordinate analysis (PCO) were used to classify the samples according to the geographical origin. Statistical analysis allowed the attribution of the analyzed populations to different chemotype groups.

Flavonoids from two Italian Genista species: Genista cilentina and Genista sulcitana

0009-3130/12/4804-0672 2012 Springer Science+Business Media, Inc. 1) Dipartimento di Chimica Scienze Farmaceutiche-sede Chimica Biorganica e Biofarmacia, Universita di Pisa, via Bonanno 33, 56126 Pisa, Italia, e-mail: cecilia.noccioli@for.pi.it; 2) Faculty of Pharmacy, Al-Zaytoonah Private University of Jordan, P. O. Box 130, Amman 11733, Jordan; 3) Dipartimento di Scienze Botaniche, Universita di Cagliari, V. le S. Ignazio da Laconi 13, 09100 Cagliari, Italia. Published in Khimiya Prirodnykh Soedinenii, No. 4, July–August, 2012, pp. 601–602. Original article submitted April 11, 2011. Chemistry of Natural Compounds, Vol. 48, No. 4, September, 2012 [Russian original No. 4, July–August, 2012]

Cytisus aeolicus Guss. ex Lindl. in vitro cultures and genistin production

Cytisus aeolicus Guss. ex Lindl. (Fabaceae family, subfamily Faboideae) is an endangered endemic species of the Aeolian Islands, Sicily. In vitro multiplication of C. aeolicus shoots was described in this work and cell cultures were established from cotyledons and hypocotyls to investigate their potential production of isoflavones. Aseptically germinated seeds, cultivated on LS modified basal medium, gave the initial explants used both to induce axillary propagation and callus cultures. The LS (Linsmaier and Skoog) basal medium, supplemented with 0.1 mg L−1 of 6-benzylaminopurine were used to induce axillary propagation. The callus induction was performed using the basal medium added with 5 mg L−1 2,4-dichlorophenoxy acetic acid and 5 mg L−1 kinetin (control medium). Basal medium was also added with 2000 mg L−1 casein hydrolysate (CH) or 900 mg L−1myo-inositol (MI). C. aeolicus callus cultures on CH and MI media produced an unique compound, the isoflavone genistein 7-O-ß-D-glucopyranoside (genistin), which has not previously been isolated from wild plants. Callus cultures grown on the medium containing myo-inositol produced the greatest amount of genistin. C. aeolicus tissue culture procedures could provide suitable plant material both for germplasm preservation (by micropropagation) and for biotechnological selective isoflavone production (by callus culture).

Antioxidant flavonol glycosides from Dorycnium hirsutum

Seven flavonol glycosides were isolated and identified from the aerial parts of Dorycnium hirsutum, together with catechin, D-pinitol, β-sitosterol, and stearic acid. The extracts and the isolated flavonol glycosides were evaluated for their antioxidant activity, using the DPPH test (radical scavenging) and the lipoxygenase assay (lipid peroxidation).

ISOLATION AND ANTIOXIDANT ACTIVITY OF FLAVONOIDS FROM Coriaria myrtifolia METHANOLIC EXTRACT

where A blank is the absorbance of the blank and A sample is the absorbance of the sample. The air-dried leaves of C. myrtifolia (30 g) were extracted with methanol. The methanolic residue was washed with n-hexane until discoloration to remove fatty acids. The polar fraction (PF1) was extracted first with ethyl acetate, yielding two fractions: ethyl acetate residue (EA; 12.5 g) and the polar fraction (PF2). The latter was extracted with n-butanol to obtain the n-butanol fraction (B; 6.15 g). The four fractions (PF1, EA, PF2, and B) were tested to evaluate their free radical scavenging activities. All fractions tested showed antioxidant activity higher than the standard (BHT, butylated hydroxyltoluene, 5.98 # 0.5 mg/mL) with IC 50 values of 1.68 # 0.5 mg/mL (B), 2.45 # 1.5 mg/mL (PF2), 2.81 # 1.0 mg/mL (PF1), and 3.68 # 1.26 mg/mL (EA). Fraction EA and B were further purified by repeated chromatographies (Sephadex LH 20, SPE, and HPLC in RP8 reverse phase) to obtain four pure flavonol glycosides: kaempferol 3-O-rhamnoside (1, 3.7 mg), quercetin 3-O-rhamnoside (2, 23.2 mg), quercetin 3-O-galactoside (3, 24.5 mg), and quercetin 3-O-glucoside (4, 11.4 mg), known as afzelin, quercitrin, hyperoside, and isoquercitrin, respectively. All these constituents gave yellow-orange fluorescence at 366 nm on TLC after spraying with NTS-PEG reagent. The identification of compounds 1‐4 was established by comparing their 1 H and 13 C NMR chemical shifts and proton coupling constants with those reported in the literature [10‐12]. These pure compounds were screened for their free radical scavenging properties according to the method used for the extracts. The IC 50 of the pure isolated flavonol glycosides 1‐4 ranged between 1.98 # 1.0 mg/mL of kaempferol 3-O-rhamnoside (1) to 3.52 # 1.0 mg/mL of quercetin 3-O-galactoside (3), with similar intermediate values of compounds 4 and 2 (2.45 # 1.5 and 2.68 # 1.0 mg/mL, respectively).

HPLC-DAD-MS ANALYSIS AND ANTIVIRAL ACTIVITY OF DIFFERENT EXTRACTS AND ISOLATED CONSTITUENTS FROM Bituminaria bituminosa

Herpes simplex virus (HSV) is one of the most common viral diseases in humans. Herpes simplex virus 1 (HSV-1) and Herpes simplex virus 2 (HSV-2) have been distinguished by clinical manifestations and biological and serological criteria. Several drugs have proved useful in the treatment of many specific infections, but viral strains resistant to these drugs have been increasingly identified [1]. In order to find less toxic antiviral agents, many researchers have addressed plant products. Indeed, many plant extracts and their secondary metabolites are known in the literature for their antiviral activity [2, 3]. Bituminaria bituminosa L. (syn. Psoralea bituminosa L., Leguminosae) is a perennial herb widespread in the Mediterranean area, known in Italy as “trifoglio bituminoso”; the fresh leaves are used in folk medicine as vulnerary, cicatrizing, and disinfectant agent [4, 5]. Plants of the genus Bituminaria are known as source of furanocoumarins and flavonoids [6, 7]. In our previous phytochemical study [8], three pterocarpans (erybraedin C and the new bitucarpins A and B) together with plicatin B were isolated and identified from B. bituminosa aerial parts. Erybraedin C and bitucarpin A showed cytotoxicity in different cell lines [9], induced apoptosis in colon carcinoma cell lines, and exhibited anti-clastogenic activity in lymphocytes [10, 11]; erybraedin C showed inhibitory activity against human topoisomerase I [12] and significant antioxidant activity [13]. The antiviral activity of the pterocarpan compounds, isolated from different plant sources, has been reported in the literature [14, 15], and some of these derivatives were active against HSV-2 [16]. In the present investigation, the antiviral activity of a few B. bituminosa extracts and their isolates erybraedin C (1), bitucarpin A (2), and plicatin B (3) is reported for the first time. The tested extracts were characterized in their quantitative chemical composition by HPLC-DAD-MS analysis.