PL231813B1 - Method for obtaining 7-O-β-D-glucopyranosyl-5,4'-dihydroxyflavone - Google Patents

Description

Description of the invention

The subject of the invention is a process for the preparation of 7-O-1 -D-glucopyranosyl-5,4'-dihydroxy flavone of the formula 2 shown in the drawing.

This compound is biologically active and can be used in industry, food and pharmaceuticals.

The literature knows 7-0 - ^ - D-glucopyranosyl-5,4'-dihydroxy flavone naturally occurring e.g. in chamomile [Vanda Śvehlikovaa, Richard N. Bennettb, Fred A. Mellonb, Paul W. Needsb, Sonia Piarentec, Paul A Kroonb, Yongping Baob, Isolation, identification and stability of acylated derivatives of apigenin 7-O-glucoside from chamomile (Chamomilla recutita [L.] Rauschert), Phytochemistry, 2004, 65: 2323-2332] and artichoke [Katrin Schutz, Dietmar Kammerer, Reinhold Carle, and Andreas Schieber, Identification and Quantification of Caffeoylquinic Acids and Flavonoids from Artichoke (Cynara scolymus L.) Heads, Juice, and Pomace by HPLC-DAD-ESI / MS ⁿ , Journal of Agricultural and Food Chemistry, 2004, 52: 4090-4096].

There is a known method of obtaining 7-0 - ^ - D-glucopyranosyl-5,4'-dihydroxy flavone by three-stage chemical synthesis, in which the starting substrate is naringenin with an efficiency of 62% [Kin-ichi Oyamaa and Tadao Kondo, Total synthesis of apigenin 7,4'-di-O-3-glucopyranoside, a component of blue flower pigment of Salvia patens, and seven chiral analogues, Tetrahedron, 2004, 60: 2025-2034].

The essence of the invention is a method of obtaining 7-O - N-D-glucopyranosyl-5,4'-dihydroxyphlavone (consisting in that the substrate, which is 4 ', 5,7-trihydroxiflavone, is subjected to microbiological transformation, as a result of which 7-0 - ^ - D-glucopyranosyl-5,4'-dihydroxy flavone Filamentous fungi of the Absidia coerulea species are multiplied, in a liquid, microbial medium, with continuous mixing of the reagents, at a temperature of 12-40 ° C. Then, it is added to the grown culture. the starting material is removed and the process is continued until the starting material is completely consumed After the transformation is complete, the transformation solution is extracted with a water-immiscible organic solvent, the organic fraction is separated, dried over anhydrous magnesium sulfate, the solvent is evaporated and the crude product thus obtained is purified by chromatographic techniques.

Preferably the fungus is the strain Absidia coerulea AM93.

It is also preferred that the reaction is carried out at a temperature of 26 ° C.

By following the invention, a glucosylation reaction in the substrate takes place as a result of the enzyme system contained in the living cells of Absidia coerulea.

The main advantage of the invention is to obtain, under mild conditions, 7-O -N-glucopyranosyl5,4'-dihydroxy flavone as the main reaction product in Absidia coerulea culture. The reaction yield is over 8%.

The invention is explained in more detail in an embodiment.

EXAMPLE 1 To a flask of 300 cm ^3, which is 100 cm ^{3 of} sterile medium containing 3 g of glucose and 1 g aminobaku per 1 dm ³ of distilled water is introduced into filamentous fungus Absidia coerulea AM93. After 6 days, the growth of microorganisms at 26 ° C with continuous shaking, is added 15 mg of 7-0 - ^ - D-glucopyranosyl-5,4'-dihydroksyflawonu of formula 1, dissolved in 1.5 cm ^{3 of} dimethyl sulfoxide. The transformation is carried out under continuous shaking for 10 days. After this time, the culture is acidified with 1-molar hydrochloric acid to pH 4.5. Then, the resulting transformation solution was extracted three times with ethyl acetate, dried with anhydrous magnesium sulfate, and the solvent was evaporated. This gives 3 mg of crude extract which is purified by chromatography using 9: 1 by volume chloroform: methanol as eluent. After purification, 2 mg of 7-O-6-D-glucopyranosyl-5,4'-dihydroxy flavone of formula II are obtained in a yield of 8.33%.

The obtained product is characterized by the following spectral data:

¹ H NMR (600 MHz, DMSO-d6) δ: 3.18 (1H, m, H-4), 3.26 (1H, m, H-2), 3.30 (1H, m, H-3 "), 3.44 (1H, m, H-5"), 3.48 (1H, m, Ha-6), 3.71 (1H, m, Hb-6), 5.07 (1H, d "J = 7.8 Hz, H -1), 6.44 (1H, m, J = 2.1 Hz, H-8), 6.83 (1H, m, J = 2.1 Hz, H- 6), 6.87 (1H, s, H-3), 6.94 (2H, m, J = 8.6 Hz, H-3 ', H-5'), 7.96 (2H, m, J = 8.6 Hz, H-2 ', H-6');

¹³ C NMR (150 MHz, DMSO-d6) δ: 60.59 (C-6 "), 69.54 (C-4), 73.10 (C-2"), 76.43 (C3), 77 , 17 (C-5 "), 94.83 (C-6), 99.51 (C-8), 99.89 (C-1), 103.11 (C-3), 105.33 (C -10), 116.00 (C-3 ', 5'), 121.02 (C-1 '), 128.62 (C-2', 6 '), 156.94 (C-5), 161 , 10 (C-9), 161.37 (C-4 '), 162.95 (C-7), 164.25 (C-2), 182.00 (C-4).

Claims (3)

Patent claims Method for the preparation of 7-O - 4 - D-glucopyranosyl-5,4'-dihydroxy flavone, characterized in that by means of a microbial transformation reaction of the substrate, which is 4 ', 5,7-trihydroxiflavone of the formula 1, is obtained by 7 -0 - ^ - D-glucopyranosyl-5,4'-dihydroxy flavone of the formula 2 in such a way that the Absidia coerulea species are multiplied in a liquid microbial medium, characteristic of fungi, with constant mixing of the reagents at a temperature of 12- 40 ° C, then after 3 to 7 days, the substrate is added to the grown culture and the process is continued until the substrate is completely consumed, then, after completion of the transformation, the transformation solution is extracted with an organic solvent immiscible with water, the fraction is separated the organic mixture is dehydrated, the solvent is evaporated and the crude product thus obtained is purified by chromatographic techniques to give pure product which is 7-O-6-D-glucopyranosyl-5,4'-dihydroxy flavone. 2. The method according to p. The method of claim 1, wherein the fungus is Absidia coerulea AM93 strain. 3. The method according to p. The process of claim 1, wherein the process is carried out at a temperature of 26 ° C.