Maciej Włodarczyk

Immunomodulatory and anti-inflammatory activity of selected osthole derivatives.

I From osthole [7-methoxy-8-(3-methyl-but-2-enyl)-chromen-2-one] (), obtained by selective extraction of Peucedanum ostruthium (L.) W. Koch roots, ostholic acid (II) was synthetized as a result of its oxidation with chromium trioxide. From ostholic acid, through its chloride, four amides were obtained: the morpholide 1, the p-chloro-benzylamide 2, the piperidine 3 and the N-methyl-piperazide 4. Except for 1, other compounds have not been described before. The amides 1 - 4 and their precursor osthole (I) were tested for their potential activities in selected immunological assays. The compounds showed moderate inhibitory activity in the humoral immune response to sheep erythrocytes in mice in vitro, and 4 was the most suppressive. The effects of 1 and 3 on concanavalin A- and pokeweed mitogen-induced mouse splenocyte proliferation were inhibitory and those of 4 stimulatory. The compounds were also tested for their activity on tumour necrosis factor α and interleukin 6 production, induced by lipopolysaccharide, in cultures of rat peritoneal cells and human peripheral blood mononuclear cells. Compounds 1, 3 and 4 inhibited tumour necrosis factor α (rat cells), whereas compound 2 stimulated the production of both cytokines. Compounds 1, 2 and 3 were also strongly inhibitory on tumour necrosis factor α production in human blood cells (73, 78 and 80% inhibition at 10 μg/ml, respectively). On the other hand, 2 and 4 stimulated the interleukin 6 production (2- to 3-fold stimulation). In addition, 2 and 4 suppressed the carrageenan-induced inflammation in mice (56.5% and 68.3% inhibition, respectively). In summary, the compounds predominantly displayed suppressive and anti inflammatory activities in the investigated models

Determination of Ursolic and Oleanolic Acid in Sambuci Fructus

Elderberries are used in the preparation of pie, jelly, punch, wine, or liqueur, as well as in many herbal remedies and food supplements. Elderberry products may provide diaphoretic, diuretic, antioxidant, and immunostimulant activities that offer protection against cold and flu. Herein, we report for the first time the qualitative and quantitative evaluation of two isomeric triterpenoids isolated from Sambuci fructus. The analysis revealed that ursolic acid and oleanolic acid are present in Sambuci fructus. The average concentration of ursolic acid was ca. three times higher than the concentration of oleanolic acid. The triterpenoids were detected and quantified using chromatographic methods such as TLC and HPLC. Spectroscopic techniques, including HR‐MS and 2D‐NMR, allowed unequivocal structure determination.

TLC as a rapid and convenient method for saponin investigation

At an early stage of investigation natural compounds in plants it is essential to have suitable analytical methods available for rapid detection and characterization of the constituents present. Identification of metabolites and their levels at this early stage is also important because it enables collection of the right plant material and acts as a guide to rational and selective isolation procedures.

In vitro evaluation of the antioxidant and antimicrobial activity of DIMBOA [2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one]

Abstract The aim of this study was to evaluate the in vitro antioxidant and antimicrobial properties of the natural cyclic hydroxamic acid: 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA). Antioxidant activity of the isolated DIMBOA was examined using DPPH, FRAP and ABTS tests. It was found that DIMBOA exhibits a potent free-radical scavenging activity and a weaker iron (III) ions reducing activity. Antimicrobial activity against selected G(+), G(–) bacterial strains and against yeasts-like reference strains of fungi was investigated using disk-diffusion method. It has been shown that DIMBOA possess growth inhibitory properties against many strains of studied bacteria and fungi, such as Staphylococcus aureus, Escherichia coli as well as against Saccharomyces cerevisiae. Graphical abstract

Rapid Densitometric Quantitative Screening of the Myricitrin Content of Crude Methanolic Extracts of Leaves from a Variety of Acer Species

Thin-layer chromatography with densitometric detection has been used for qualitative and quantitative analysis of the myricitrin content of extracts of the leaves of 20 species of the Aceraceae family. The presence of myricitrin was confirmed in five of the samples. The mobile phase was optimized for quantitative analysis of myricitrin without extract purification. Myricitrin standard was obtained from one of the extracts; its identity was confirmed chromatographically and by use of spectral methods. Implications for chemotaxonomic and phytochemical research are discussed.

β-Aescin at subinhibitory concentration (sub-MIC) enhances susceptibility of Candida glabrata clinical isolates to nystatin

Aescin (escin) derived from the seeds of horse chestnut (Aesculus hippocastanum L.) is a natural mixture of triterpene saponins exhibiting a wide variety of pharmacological properties, including antiinflammatory, analgesic, and antipyretic activities. However, data concerning antifungal activities of these compounds are limited. This study aims to evaluate the in vitro antifungal susceptibility of Candida glabrata clinical isolates to α-aescin sodium, β-aescin crystalline and β-aescin sodium using the disk diffusion (DD) and broth microdilution (BMD) methods. Moreover, the influence of subinhibitory concentration (0.5×MIC) of β-aescins on the nystatin MIC was also studied. In general, the results obtained by the DD assay correlated well with those obtained by the BMD method. Both β-aescins effectively inhibited the growth of all 24 strains tested. The minimum inhibitory concentration (MIC) values ranging from 8 to 32 μg/ml for β-aescin crystalline, whereas those of β-aescin sodium were slightly lower and ranged from 4 to 16 μg/ml. In contrast, α-aescin sodium was found to be completely ineffective against the strains studied. MIC values of nystatin were reduced 2-16-fold and 2-4-fold in the presence of subinhibitory concentration of β-aescin crystalline and β-aescin sodium, respectively. Results of the present study may suggest the additive interaction between β-aescin and nystatin.

Aescin determination in seeds and capsules of three pure and hybrid Aesculus species

0009-3130/11/4701-0142 2011 Springer Science+Business Media, Inc. 1) Department of Pharmacognosy, Wroclaw Medical University, pl. Nankiera 1, 50-140 Wroclaw, Poland, fax: +48 71 7840218, e-mail: michalg@farmgn.am.wroc.pl; 2) Universito Ricercatore di Biologia Farmaceutica Dipartimento di Scienze Farmaceutiche Universita Degli studi di Salerno, via Ponte don Melillo, 84084-Fisciano, Italy. Published in Khimiya Prirodnykh Soedinenii, No. 1, p. 126–128, January–February, 2011. Original article submitted September 9, 2009. Chemistry of Natural Compounds, Vol. 47, No. 1, March, 2011 [Russian original No. 1, January–February, 2011]

A Major Saponin from Leaves Extract of Acer velutinum

A major triterpenoid saponin was isolated from the leaves extract of Acer velutinum. This compound was characterized as 21β ,22α-O-diangeloylprotoaescigenin 3-O-[β -D-glucopyranosyl- (1→2)][β -D-glucopyranosyl-(1→4)]-β -D-glucuronopyranosyl acid. Its structure was elucidated by 1H, gCOSY, ROESY, gHSQC, gHMBC NMR as well as ESI-MS experiments. Isolated saponin exhibited in vitro cytotoxicity against HL-60, B16-F0 and BALB/3T3 cell lines. Graphical Abstract A Major Saponin from Leaves Extract of Acer velutinum

Nitrogen compounds in Phacelia tanacetifolia Benth. honey: First time report on occurrence of (−)-5-epi-lithospermoside, uridine, adenine and xanthine in honey

Lacy phacelia (Phacelia tanacetifolia Borkh.) honey composition was screened by UHPLC-DAD-QqTOF-MS. The targeted analysis revealed 6 major nitrogen compounds including aromatic amino acids (tyrosine, phenylalanine), purine derivatives (adenine, xanthine), nucleoside (uridine) and rare non-cyanogenic cyanoglucoside, (-)-5-epi-lithospermoside ((2Z)-2-[(4R,5R,6S)-4,5-dihydroxy-6-(β-d-glucopyranosyl)oxycyclohex-2-en-1-ylidene]acetonitrile). Their identity was confirmed by different analytical tools: HRMS, co-chromatography with standard compound or comprehensive NMR experiments. All the compounds, except amino acids, were reported and determined in honey for the first time. The amount of the compounds was quantified in 16 unifloral phacelia samples: adenine (18.45 ± 4.63 mg/kg), xanthine (10.53 ± 2.98 mg/kg), uridine (42.84 ± 9.26 mg/kg), tyrosine (14.66 ± 10.22 mg/kg), (-)-5-epi-lithospermoside (70.61 ± 31.37 mg/kg) and phenylalanine (20.41 ± 11.99 mg/kg). The (-)-5-epi-lithospermoside content is significantly correlated with P. tanacetifolia pollen percentage (R2 = 0.5612, p < 0.001) and it is proposed as a potential marker of botanical origin for phacelia honey.

Triterpenoid Acids as Important Antiproliferative Constituents of European Elderberry Fruits

ABSTRACT In Europe, both the fruits and flowers of Sambucus nigra L. have been used against cold, as well as laxative, diaphoretic, and diuretic remedies. There are also a number of commercially available food products that contain elderberry juice, puréed or dried elderberries. Recent comprehensive literature data on pharmacology and chemistry of Sambuci fructus have encouraged us to screen extracts with different polarities from this plant material against cancer cell lines. The cytotoxic activity of the ethyl acetate and aqueous acetone extracts from elderberries as well as detected triterpenoids on human colon adenocarcinoma cell line (LoVo) and human breast cancer cell line (MCF-7) was investigated by sulforhodamine B assay. Moreover, cell migration assay was conducted for triterpenoid fraction and pure compounds. Aqueous acetone extract possessed much lower IC50 value in cancer cell lines compared to ethyl acetate extract. The latter manifested high cytotoxicity against studied cell lines, suggesting that nonpolar compounds are responsible for the cytotoxic activity. Indeed, the phytochemical analysis revealed that ursolic and oleanolic acids are the main triterpenoids in the mentioned extract of which ursolic acid showed the highest activity with IC50 values of 10.7 µg/mL on MCF-7 and 7.7 µg/mL on LoVo cells.