LU81679A1 - 3,4-DIHYDROXYCIMALDEHYDE DERIVATIVE, THEIR PRODUCTION AND MEDICINAL PRODUCTS - Google Patents

Description

- 5 - 3,4-Dihydroxycinnamaldehyde derivative, its manufacture and pharmaceuticals

The invention relates to the compound of the formula!

Η v L JL i ..CH, OH

r '0 CHo0H I

(c oier T

/ \ ^ oh

0 = C H

and their salts. This connection is called "American".

5 This compound has a surprisingly favorable anti-hepatotoxic activity, which is a welcome addition to the spectrum of activity i of the known liver drugs.

You get this new connection z. B. by extraction and fractionation from fruits of the Phytolacca species, in particular 10 of the Phytolacca americana L., by extracting finely ground dried fruits of Phytolacca americana L. after treatment with petroleum ether with acetone, cleaning the residues of the evaporated extract with chloroform and dissolves in n-butanol, the n-butanol solution is purified with water, evaporated and 15 the residue is dissolved in acetone, the acetone solution is chromatographed on / 7 - 6 - r • Sephadex LH 20 with acetone as eluent in a column which pharmacologically virksaaen fractions combined and evaporated and the residue tm-crystallized from methanol.

5 The procedure is preferably such that a) finely ground, dried fruits of Phytolacca americana L. are three times 0.5 to 1.5 hours, preferably 1 hour, each 2.5 to 4.5 times, preferably the 3.5 times the amount by weight of petroleum ether (boiling point> 10 in the range 60-70 ° C.) at .45 to 55 ° C., preferably 50 ° C., with strong stirring, b) the drug pretreated in this way is treated 0.5 to 0.5 times 1.5 hours, preferably 1 hour, each with 2.5 to 4.5 times, preferably 3 to 5 times the amount by weight of acetone at 45 to 15 55 ° C., preferably 50 ° C., extracted with vigorous stirring, c) evaporate the acetone extract at 35 to 45 ° C, preferably 40 ° C, in vacuo at 5-50, preferably 20 mbar, d) the residue obtained is obtained twice with 1/4 to 1/2, preferably 1/3 chloroform to the weight of the original 20 drug and filtered, e) the remaining solid in a ratio of 1:10 bie (1:14, preferably 1:12, in n-butanol, f) the n-butanol solution 3 to Shake 5 times, preferably 4 times, against 1/3 to 2/3, preferably half the volume of 25 water, g) the n-butanol phase at 35 to 45 ° C., preferably 40 ° C., in vacuo at 5-50, preferably Evaporates 20 mbar, h) the residue is dissolved in 0.05 to 0.15 times, preferably 0.10 times the amount of acetone, based on the weight of the original drug, and filtered,. / ^ l

J

- 7 - i) the acetone solution is chromatographed on Sephadex LH 20 using acetone as eluent on a column, j) the resulting pharmacologically active fractions 5 are combined and are heated to 35 to 45 ° C., preferably 40 ° C., in vacuo at 5 - 50, preferably 20 mbar evaporates, k) and the resulting residue is recrystallized from methanol.

^ The product thus obtained is a light yellow crystalline ’10 substance with the following properties: melting point 246-7 ° C.

, -v »with decomposition, molecular formula C ^ gH ^ gOg, molar mass 328,

Elemental analysis C 64.98 ¢, H 4.98 ¢, 0 28.3 Rf-Vert 0.17 in thin-layer chromatography using Kiegelgel-60 plates Merck and chloroform / 15 acetone / formic acid 9/2/1 (V / V / 7) as a flow agent system, practically insoluble in water, chloroform, benzene and petroleum ether, soluble in dimethyl sulfoxide, dimethylformamide, tetrahydrofuran, dioxane and acetone, optical rotation Mi7 of + 23.7 ° and IR absorption at Λ ** 4 20 3200 cm (-0H), 1650 a (e6, ß-unsaturated C = 0), 1610 cm-1 (= CC) and 1580, 1510 and 1450 cm-1 (aromatic system).

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The IR and mass spectrum of the product is shown in Figs. 1 and 2.

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- 8 - i

This new compound can also be obtained in the following way, that 3 »4-dihydroxycinnamaldehyde and 3» 4-dihydroxycinnamyl alcohol are dimerized by means of oxidative addition.

The procedure is preferably such that 'equimolar 5 amounts of 3i4-dihydroxycinnamaldehyde and 3' 4-dihydroxycinnamon alcohol in 5 times the amount of the total reagents of a solvent such as benzene, dioxane, tetrahydrofuran or acetone at 15 to 30, preferably 23 ° C can react for 1.5 to 3 »preferably 2 hours, under the action of silver oxide 10 in a ratio of 0.5 to 0.6, in particular 0.55 mol, per 1 mol of total reagents, which brings the filtered reaction solution to dryness and the residue is separated off by column chromatography.

The product obtained synthetically for this has the same 15 characteristics as that obtained by isolation from the phytolacca

Types of product obtained.

This new compound can also be obtained in the following manner by reacting 3 '4-dihydroxycinnamaldehyde with allyl catechol in an organic solvent by means of oxidative addition, acetylating the resulting intermediate, separating it by column chromatography, oxidizing the (methyl substituent in the benzodioxane group to the alcohol function and The compound obtained is saponified with a weak alkali, 25 preferred procedures are carried out in such a way that equimolar amounts of 3 »4-dihydroxycinnamaldehyde and allyl catechol are added in 5 times the amount of the total reagents of a solvent vie / - 9 -

Benzene, dioxane, tetrahydrofuran or acetone hot 15 dis 50 ° 0 "preferably 25 ° C, during 1.5 to 3" preferably 2 hours, under the action of silheroxide in a ratio of 0.5 to 0.6, especially 0.55 Mol per mole of total reagents, can react, 5 the product obtained acetylated in known Veise, isolated by column chromatography and then oxidizing the methyl substituent on the benzodioxane group to alcohol function in known Veise.

The product obtained synthetically for this has the same 10 characteristics as the product obtained by isolation from the Phytolacca species.

(0 ^ J§X0H + W0H - 2 V ^^ OH 'HK in benzene

v Q μ with trace CHjOH

H Allylhrenzcatechin 3,4-Dihydroxyzimtaldehyd 0 v. · ’^^ 3 Γ / ^ νΤ j Oxidation - *

After acetylation and preparation, S.C, isolable ov-CH2oh

> 0 toT

Ajnericapin / - 10 -

A first structure elucidation of the compound according to the invention has the following formula! jV ”tr JL JL JL ___, ch2oh ^ c> -v \ h2oH hrv„ «C or UQf • 0 = C / XH ^

This connection is called "American".

This substance can be converted into its physiologically tolerable salts in the usual way. Weakly basic agents suitable for salt formation are, for example, alkali and alkali hydrogen carbonates and amines such as triethanolamine, trimethylamine, triethylamine, aminosugar, etc.

[The product according to the invention can be used both as a unit tract and as a crystalline product for medical purposes

Liver therapy can be used. It can be used in various pharmaceutical forms such as tablets, capsules, granules, coated tablets, suppositories or as a liquid preparation orally, anal or parenterally. The dose of dumand is 15 200 - 450 mg / day, depending on the severity of the case.

.U- · - 11 -

In the described active ingredient Americanin and its salts, surprisingly "in animal experiments, a pronounced protective effect against liver-damaging influences was found. Americanin has a particularly strong protective and stabilizing effect on the cellular and intracellular biomembranes, particularly on the liver cells, ie a strong liver protection effect and can therefore be used, for example, in the treatment of liver diseases.

To demonstrate the antihepatotoxic activity of the product according to the invention, female mice were damaged with the liver poison '* phalloidin (3 mg / kg; i.p.) (control animals) and the

Test animals additionally treated with Americanin.

The Americanin was dissolved in DMSO + polyethylene glycol (lsl), i.v. administered, namely in batches of 50 hzw. 100 mg / kg 15 hours before the phalloidin injection.

Resulti

Of the mice poisoned with phalloidin, 85 died while the tades rate by treatment with 50 hzw. 100 mg / kg Americanin to 60 hzw. 30 / £ could be reduced (observation period: ^ 20 7 days; number: 20 animals per group).

The toxicity (i.v.) of the new active ingredient according to the invention was hei male hzw. female mice examined. Values of 820 mg / ^ tg (for male mice) and 800 mg / kg (for female mice) were found.

25 The production of the product according to the invention is described below using an explanatory example:

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EXAMPLE 1 100 kg of finely ground, dried fruits of Phytolacca americana L. are extracted three times for 1 hour each with 400 l of petroleum ether (boiling range 5 60-70 ° C.) at 50 ° C. using an Ultra-Turrax device. The one so pretreated

The drug is then subjected three times to a 1-hour Turrax extraction with 400 l of acetone each at 50 ° C. The combined acetone extracts are evaporated in vacuo at 40 ° C. The residue obtained is then stirred twice with 50 1 10 chloroform. The mixture is filtered and the residual solid dissolved in 1:12 in n-butanol. The n-bntanol solution is shaken out four times with half the volume of water. The butanol phase is then evaporated in vacuo at 40 ° C. and the residue is dissolved in 15 10 1 acetone. After the undissolved portions have been separated off, the solution is chromaiographed on Sephadex LH 20 using acetone as eluent in a column.

The fractions containing the active ingredient are combined and evaporated at 40 ° C. in vacuo. The residue is recrystallized from methanol.

The end product is a substance with the following chemical and physical parameters:

Appearance: light yellow microcrystalline solid solubility; soluble in dimethyl sulfoxide, dimethylformamide 25 tetrahydrofuran, dioxane and acetone, slightly soluble in methanol, ethanol and butanol, practically insoluble in water, chloroform, benzene and petroleum ether

Melting point: 246 - 7 ° C (decomposition) '/ _ - 13 - Sïumnenf ornel t ^ 18 ^ 10 ^ 6

Molar mass: 328

Elemental analysis: C 64.98 (calc. 65.85) H 4.98 (calc. 4.9l) 4 0 28.3 (calc. 29.24)

Thin layer chromatography:

Ready-made silica gel plates (Merck)

Eluent: chloroform, acetone, formic acid 9/2 / l (V / V / V) development: under chamber saturation 10 detection: spray reagent 2,4-dinitrophenylhydrazine / sulfuric acid (lg 2,4-dinitrophenylhydrazine in 2 ml concentrated sulfuric acid and slurried with methanol made up to 100 ml)

After spraying the plate, it is heated to 15 120 ° C. for 20 minutes. The substance appears as an intense yellow zone on a light yellow background.

Ef-Vert: 0.17

The IR spectrum and mass spectrum are shown in the attached Figures 1 and 2.

/ / i - 14 - r '.

Galenic examples:

Herst e 1 lung_y ° n_Tabl e tt en: 70 kg of the active ingredient according to the invention are mixed with the following auxiliaries: 5 10,000 kg of polyvinylpyrrolidone 14,400 kg of microcrystalline cellulose 17,400 kg of Amylum tritici 6,500 kg of aerosil 10,000 kg of stearic acid 10 571,700 kg of lactose DIN 80

Then tablets of 0.50 g (containing 70 mg of active ingredient) are pressed.

Manufacture of injectables (ampoules);

To produce 10,000 ampoules, 1.5945 kg of methyl 15-glucaain salt of the active ingredient according to the invention are dissolved in 48.6 liters of physiological saline solution, to which 4 polyvinylpyrrolidone (M.G. ^ vIO.OOO) are added. The pH should not fall below 7.6. The solution is sterile filtered and filled into sterile brown 5 ml ampoules, so that the content per ampoule is 159.45 Hg N-methylglucamine salt, corresponding to * 100 mg of the active ingredient according to the invention.

Manufacture of suppositories: 318.9 g of the methylglucamine salt of the active ingredient according to the invention are ground with 500 g of melted hard fat DASS 7 25 While stirring, 1,181.1 g of melted hard fat DAB 7 are added and suppositories are poured from the mass. Each suppository of 2.0 g contains 318.9 mg of methyl g lue amine salt, corresponding to 200 mg of the active ingredient according to the invention.

/ '/ Ί * 4

Claims (6)

1. Compound of the formula rt '/ X I ^ .ch.oh c 0 CM «y 2 ° the' LA ... 0 = C H. 0H H and their salts. 2. The method according to claim 1, characterized in that a) extracted either finely ground dried fruits of Phytolacca americana L. after treatment with petroleum ether with acetone, the residues of the evaporated extract are cleaned with chloroform and dissolved in n-butanol, the n -Butanol solution is purified with water, evaporated and the residue is dissolved in acetone, the acetone solution is chromatographed on Sephadex LH 20 with acetone as eluent in a column, the pharmacologically active fractions are combined and evaporated and the residue is recrystallized from methanol, or i / -vf - 2 - r: b) 3> 4 “Dibydroxyzimtaldebyd and 3» ^ Dihydroxyzimtalkohol dimerized by means of oxidative addition, or c) 3,4-Dihydroxyzimtaldehyd reacted with allylpyrocatechol in an organic solvent by means of oxidative addition, the intermediate obtained acetylated, separated by column chromatography, the methyl substituent in the benzodioxane group to give Alk oho 1 fu nkt ion oxidized and the compound obtained saponified with a weak alkali. (' ' 1 3. The method according to claim 2a), characterized in that a) finely ground, dried fruits of Phytolacea americana L. three times 0.5 to 1.5 hours, preferably 1 hour, each with 2.5 to 4.5 times, preferably the 3i5 times the amount of petroleum ether (boiling range 60-70 ° C) at 45 to 55 ° C, preferably 50 ° C, extracted with vigorous stirring , b) the drug pretreated in this way three times 0.5 to 1.5 hours, preferably 1 hour, each with 2.5 to 4.5 times, preferably 3.5 times the amount by weight of acetone at 45 to 55 ° C., preferably 50 ° C, extracted with vigorous stirring, c) evaporating the acetone extract at 35 to 45 ° C, preferably 40 ° C, in vacuo at 5-50, preferably 20 mbar, d) the residue obtained twice with l / 4 to 1/2, preferably 1/3 chloroform based on the weight of the original drug, and filtered, e) dissolving the remaining solid in a ratio of 1:10 to 1:14, preferably 1:12, in n-butanol, L * - 3 - f) the n-butanol solution is shaken 3 to 5 times, preferably 4 times, against 1/3 to 2/3, preferably half the volume of water, g) the n-butanol phase at 35 to 45 ° C., preferably 40 ° C, evaporated in vacuo at 5-50, preferably 20 mbar, h) the residue in 0.05 to 0.15 times, preferably. 0.10 times the amount of acetone, based on the weight of the original drug, dissolved and filtered, i) the acetone solution was chromatographed on Sephadei LH 20 using <acetone as eluent in a column », j) the pharmacologically active obtained Fractions combined and evaporated at 35 to 45 ° C, preferably 40 ° C, in vacuo at 5-50, preferably 20 mbar, k) and the resulting residue recrystallized from methanol. '4. Process according to claim 2b), characterized in that equimolar amounts of 3t4-dihydroxycinnamaldehyde and 3,4-dihydroxycinnamyl alcohol are 5 times the total amount of the total reagents of a solvent such as benzene, dioxane, tetrahydro-furan or acetone at 15 to 50 , preferably 23 ° C for 1.5 to 3, preferably 2 hours, under the action of silver oxide in a ratio of 0.5 to 0.6, in particular 0.55 mol, per mole of total reagents, the filtered reaction solution dries to dryness brings and the residue separated by column chromatography. ί / v! /; a ♦ - 4 - * 5. The method according to claim 2c), characterized in that apiimolar amounts of 3> 4-dihydroxycinnamaldehyde and allyl catechol in the 5-fold amount of the total reagents of a solvent such as benzene, dioxane, tetrahydrofuran or acetone at 15 to 30 ° C, preferably 25 ° C, during 1.5 to 31 preferably 2 hours, under the action of silver oxide in a ratio of 0.5 to 0.6, in particular 0.55 mol, per mol of total reagents, react, the product obtained acetylated in a known manner, isolated by column chromatography and then the methyl substituent on the benzodioxane ('group in the alcohol function is oxidized in a known manner. 6. Medicament, characterized by a content of compound according to claim 1, usual carriers and / or diluents. \ ί '· "r \ Λ. A 1 \ f« »