FI63418C - FOERFARANDE FOER TILLVARATAGANDE AV REN KRISTALLIN BETA-AESKINUR FROEN AV HAESTKASTANJER - Google Patents

Description

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Patent neridelat - (51) Kv.ik? /Int.a. C 07 G 3/00, C 07 J 63/00 ENGLISH - EN N LAN D (21) hteotdhikemu · —NtwdwekiHi »g 761957 (22) HakMnltpiivI —AMeimInfAg O5.O7.76 '(13) Alkupllvl — GUtightMdig O5. O7.76 (41) Tullut JulkiMkd - Blivlt j_2 01 77

Patent and Register Holders (44) NihUvUutpwton j «heardJuUuiMn and others -

Patent · och regitteratyrelien AmMum uthifd oeh utLskrtfun publkwrad 28.02.83 (32) (33) (31) Pjnrftny Muoik «u-4« «ftrd prtorlut 11.07-75

Federal Republic of Germany-Förbundsrepubliken Tyskland (DE) P 253091 + 1.5 Proof-Styrkt (71) Dr. Madaue & Co., Postfach 93 20 01, 5 Cologne 91 (Merheim),

Federal Republic of Germany-Förbundsrepubliken Tyskland (DE) (72) Rolf Madaus, Cologne-Briick, Federal Republic of Germany-Förbundsrepubliken Tyskland (DE) (7 ^) Leitzinger Oy (5 ^) Procedure for recovering pure crystalline The present invention relates to a process for recovering pure crystalline 8-aeskin (specific rotation -27 °) from seeds of horse chestnut (Aesculus hippoeastanum) which are extracted with an aqueous solution of the lower alkyl alcohol. with, the resulting eluate is evaporated by distillation after removal of the cation exchanger, water is added, the resulting crystalline precipitate is removed by filtration or centrifugation.

Until now, it has been hypothesized that the crystallizable acidic saponin is bound to the herb via hitherto unknown groups and can be dissolved out of this natural or native complex by weak acid hydrolysis. Although the water solubility of the native saponin complex is still satisfactory at approximately neutral pH, the water solubility of aeskin released therefrom is very low.

U.S. Pat. No. 3,238,190 discloses an extraction method described in the introduction for recovering a crystalline escin mixture from horse chestnut seeds. The sesquin mixture contains both the ά and 2,641 8/3 isomers. However, it has now been found that the pharmacological effect of the o <-isomer of aeskin is rather small, which adversely affects the overall effect of the aeskin mixture.

With the process according to the invention, pure β-aescin can now be recovered in high yield. The process according to the invention is characterized in that the weight ratio of extractable to solvent is about 1:10 and the extraction solvent is a methanol / water mixture containing 65% by volume of methanol, that the extraction is carried out by vortex extraction, the extracts are to a pH of about 3.5, not more than 3.8, the crude aeskin is dissolved in methanol, treated with activated carbon and mixed from the methanol solution by adding water, and the resulting aesic acid is converted, if necessary, with the pharmacologically acceptable base into the corresponding salt.

The composition of the aeskin obtained by the process according to the invention is always the same in percentage, as evidenced by the numerous batches carried out and confirmed by X-ray analysis, NMR spectrography, chromatographic separation, optical rotation measurement and hemolytic index determination. In the process of the invention, the defatted and finely ground drug raw material is mixed with a mixture of methanol and water and extracted using vortex flow extraction at room temperature. The mixture ratio is 1 part of drug raw material per 10 parts of extraction liquid and the extraction time is 15 minutes. In beakers and separators, the drug raw material is separated from the primary extract. The primary extract is then filtered using polished filters. The ion exchange then takes place, in an appropriate manner, so that, after the column process, a strongly acidic cation exchanger is used in its H + form. Flow occurs from the bottom up through the column. The pH of the escaping escapine solution must not exceed 3.5. Regeneration of the ion exchanger takes place with 10% aqueous hydrochloric acid at 70 ° C.

The aesquin solution passed through the ion exchanger is evaporated to dryness by distilling off the solvent, so that the evaporation ratio is between 2: 1 and 4: 1 and preferably 2.5: 1, which means that the solution containing 10 liters of aeskin is evaporated to 4 liters. the raw aeskin crystallizes out. After removal of the crude aeskin 6341 8, it is washed with hot water, centrifuged once more and dried in a vacuum oven at 80 ° C.

For recrystallization, the crude sesame is dissolved in 10 times the amount of methanol. 5% of ground activated carbon, based on the amount of crude aescin, is added to the mixture, after which the solution is heated to boiling under a vertical condenser, filtered and evaporated to half volume. After repeated filtration, twice the amount of mineral water is added to the metaphiol solution at 90 [deg.] C. and, after cooling at room temperature, the crystalline escine separated is separated by a chamber separator and dried in vacuo at 80 [deg.] C., the yield is about 1.5-2.0%. degree of purity 97 - 100% acein.

The resulting esquin can be converted to a salt in solution with a suitable base, care being taken to ensure that the pH does not exceed 7 on neutralization. An inefficient esquinol is formed in the alcoholic solution. Preferred salt-forming agents are, for example, alkali metal bicarbonates, which are added in solid form to an alcoholic sesquin solution. In addition, mono-, di- and trialkanolamines, especially triethanolamine, are preferred.

The resulting acein has a pharmacologically selective anti-edematous, anti-swelling, anti-inflammatory and antithrombotic effect. It can be used in the usual pharmaceutical preparations.

The acid form is sparingly soluble in water, as a result of which it is preferably converted into the form of an alcoholic solution or gel 6341 8 4 for use as a topical active ingredient. The salts, especially the sodium salt, are highly soluble in water and can be used as tablets, granules, solutions, gels, ampoules. The oral dosage, calculated as acidic sesquin, is 60 to 120 mg. For example, the intravenous dosage of sodium sesquinate averages 5 to 10 mg per day calculated as acidic sesquine.

Acute toxicity figures (DLmin - DL ^ q) vary depending on the animal species and experimental conditions (breed, mode of treatment, nutritional status). Following intravenous administration, the following values were obtained (all figures are in mg / kg): mouse: 1.5 to 9.3 rat: 2.0 to 16.8 guinea pig: 7.5 to 9.1 rabbit: 3.0 to 5.0 pig: 4.0 dog: 3.0

Examination of chronic toxicity in animal experiments did not show any pathological changes in the animals. Accurate studies on foetotoxicity and malformation have shown that aescin does not cause any malformations.

The pharmacological effects of aeskin have been studied in more than 30,000 individual trials. The results obtained provide a broad and secure basis for its therapeutic use.

5 6341 8

More specifically, for example, the effect of aeskin on inflammatory edematous conditions - from mere oozing to severe tissue necrosis - has been studied, which can be provoked by the following substances: ovalbumin, dextran, local arthus phenomenon, carrageenan, hyaluronidase, bradykinin, compound 48/80 , aerosol, kaolin, bee venom, baker's yeast and formalin. In all cases, the experimental model was a well-measurable rat paw edema. Most clearly, there are significant effects on edema in which the infiltration of blood fluid into the extracapillary region is characteristic. In addition to paw edema, aeskin can also prevent guinea pig pulmonary edema provoked by ovalbumin or histamine. Very comparable to human conditions is traumatic swelling of the rat paw through local, artificial hemorrhage.

Good results have been obtained with aeskin in the treatment of spinal pain syndromes. Vertebral-related pain conditions, shoulder and arm syndrome (zervikal syndrome), LVS syndrome (lumbago, sciatica, etc.) have been the subject of increasing research. Use of acein, leaves. that the occurrence of venous blood arrest in dilated venous mopunus in the area of the spinal canal has originally been involved in pathological events. The so-called radical edema and the effect of venous blood flow disorders on the elementary scaffold can be eliminated with the help of aeskin.

In more than 800 cases, it has been found that even pain conditions that cannot be cured with continuous treatment, which are accompanied by considerably large limitations of spinal mobility combined with nerve root irritation, respond surprisingly well to treatment with aeskin.

Compared to the usual treatment methods so far, a significant shortening of the treatment period and a significantly better improvement rate (90%) have been observed. It is expedient for the treatment to be carried out for at least one week with ampoule ampoules (sodium escinate 5 mg) as a daily treatment and, depending on the state of recovery, thereafter 2-3 times a week for 1 ampoule at a time. Aeschine granules can be added to the treatment afterwards or the treatment can later be switched entirely on them to achieve the results of the treatment.

In brain dysfunction due to edema, including acute and life-threatening cases, aeskin has been shown to be very useful 6341 8 6.

Aeskin has been widely used in numerous accidental brain injury conditions (commotation, contusion) that occur in traffic accidents.

When accidents are cured, it is substantially easier to remove post-accident and post-operative edema from the patient. Aeskin is then suitable for healing wounds, fractures, dislocations, sprains and bruises.

The use of pre-operative injections of 2 ampoules of sodium escinate a 5 mg, administered appropriately repeated at 12-hour intervals during the first 2-3 days, makes it possible in many cases to prevent the development of post-operative edema.

7 „. ,,. „6341 8

Examples Example 1 10 kg of seeds of the plant Aesculus hippocastanum were finely ground under a vortex flow extraction in 100 liters of a methanol-water mixture (65% by volume). Prior seeding or degreasing was not necessary. After removal of the drug raw material by decantation and separators, the extract passes through a column packed with a strongly acidic cation exchanger in the H + form (eg Lewatit S 100) and preferably from the bottom upwards, giving a pec value of 3.5 for the escaping aeskin solution. chapter 3.8. The extract is then evaporated in a ratio of 10: 4 and the crystallized crude aescin is dried in vacuo at 80 ° C. The crude aescin is then dissolved in 10 times the amount of methanol, 5% powder based on the amount of crude aescin is added to the solution, and the mixture is heated under a vertical condenser. After filtration, the amount of mineral water obtained (90 ° C) is added to the hot methanol solution and the separated crystalline aeskin is removed and dried in vacuo at 80 ° C. The resulting aeskin has a melting point of 222-227 ° C, a rotation of -28 ° + 2 ° and pKs U.7.

Example 2 1000 g of aeskin, recrystallized 3 times from methanol with 5% activated carbon are dissolved in 15 liters of methyl ethyl ketone-denatured ethanol with a maximum temperature of 70 ° C with constant and efficient stirring and a stoichiometric amount of sodium hydrogen carbonate p.a. serially. During the addition of sodium bicarbonate, the temperature of the mixture must not exceed 50-60 ° C.

Salt formation in the heterogeneous phase requires 4 to 5 hours.

During this time, any added sodium bicarbonate dissolves.

After the above time, 8 l of ethanol are then removed by distillation at a maximum temperature of 50 ° C and in vacuo. The remaining solution is filtered. The resulting sodium escinate crystallizes overnight as colorless needles. The crystal porridge is removed from the mother liquor and dried in a vacuum oven at 50 ° C for 15 hours.

The free acid content of the ground product is analyzed analytically.

The analyzed sodium sesquinate is redissolved in 15 liters of ethanol denatured with methyl ethyl ketone and reacted again with the calculated amount of sodium hydrogen carbonate p.a. as described above.

In this case, it is necessary to note that the minimum alcohol content added is 96% by weight, as otherwise there could be a "risk of the alkaline reaction sympathizing with p ^ 8, resulting in the decomposition of aeskin and its conversion to aeskinol.

The solution is treated as described above. After grinding, the sodium sesquinate is post-dried at 70 ° C in a vacuum oven for 24 hours. The yield is 715 g = 70.2% of theory.

After two reactions, the product is about 97%, with about 3% acid.

Example 3 1000 g of aeskin are dissolved in 15 liters of methanol with vigorous and continuous stirring. A stoichiometric amount of sodium bicarbonate corresponding to the concentration of aesquin is added to it in solid form and at a temperature of about 50 ° C with vigorous stirring. After completion of the reaction, the resulting sodium escinate solution is transferred to spray drying in an inert gas phase (nitrogen).