Method for preparing lappaconitine hydrobromide (versions)

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, namely to a method for preparing lappaconitine hydrobromide (versions) A method for preparing lappaconitine hydrobromide is implemented by extraction of Aconitum leucostomum root and herb or Aconitum septentrionale root and herb in methylene chloride in a continuous extraction apparatus that is followed by decontamination by means of flash chromatography (version 1), or extraction of a herbal raw material in a polar organic solvent followed by extract removal from the organic solvent (version 2), alkalinisation and extraction of the prepared extract in methylene chloride followed by decontamination of the extract by flash chromatography.

EFFECT: method for preparing lappaconitine hydrobromide provides simplifying the technological process, reducing its length and improving higher yield of the end product of officinal purity.

7 cl, 1 tbl, 9 ex

 

The invention relates to chemical-pharmaceutical industry, namely to method of obtaining (and its variant) lappaconitine hydrobromide (VFS) that is used as antiarrhythmic drug funds.

Lappaconitine hydrobromide is obtained by extraction of roots or herbs Aconite beloustogo (Aconitum leucostomum) or of roots or herbs of Northern monkshood (Aconitum septentrionale).

Known methods for producing lappaconitine based on the specified extraction of vegetable raw materials with different solvents: methanol (Marion L., L. Fonzer, Wilkins C. K., Boca Jr and J. P., F. Sandberg, R. Thorsen, Linden E. // Can J. Chem. 1967. vol. 45, pp. 969-973 [1]); 80%-ethanol (Ross A., Pelletier S. W.//Tetrahedron. 1992, vol. 48, No. 7, pp. 1183-1192 [2]; dichloroethane (T. F. Platonov, Kuzovkov, A. D., Massagetae P. S.//Doha. - 1958, p. 258-261 [3] ); chloroform (Usmanov S. K., V. Telnov, A., Yunusov, M. S., Abdullaev N. Etc., Schroeter, A. I., Filippov G. B. // HPS. - 1987, pp. 879-883 [4] ).

With slight variations of extraction conditions and extractants are described in the prior art the conventional order of lappaconitine comprising the following steps:

extraction of crushed dried vegetable raw materials with an organic solvent,

- evaporation of the resulting extract to remove the solvent,

- acidification of the residue to an acidic reaction,

- extraction of the obtained acidic solution is not miscible with water org�organic solvent to remove impurities non-core nature,

- alkalinization of the aqueous solution until alkaline reaction,

the Department released lappaconitine (by filtration or extraction),

- placing the base of lappaconitine hydrobromide in the action of Hydrobromic acid,

- clean to pharmacopoeial purity of lappaconitine hydrobromide salt by crystallization from alcohol.

In some cases additionally used column chromatography on alumina for purification of the bulk of the Foundation of lappaconitine [1] or the joint of the manifold [2].

The structure of lappaconitine contains a fragment of the ester of N-acetyl Anthranilic (o-aminobenzoic acid). Therefore, this molecule is very labile, and therefore in acidic and basic environments can quickly occur hydrolysis of both ester and N-acetyl fragment. Thus, a process for obtaining lappaconitine from plant material according to known methods requires great care in keeping the solution pH and temperature at the stage of acidification evaporated extract, at the stage of alkalization after extraction of impurities non-core nature, and upon receipt of the hydrobromide acid Hydrobromic acid base lappaconitine. In addition, to avoid hydrolysis of all of these methods should be fast and not take m�wow time to minimize side hydrolysis processes. Failure to comply with the above conditions can easily lead to a significant reduction of the yield of the target product due to hydrolysis in one or more stages of the method.

The closest to the claimed method is a method of producing lappaconitine hydrobromide by 3-5-fold extraction of roots or herb Aconite North or roots or herbs Aconite beloustogo 50-80% by volume aqueous solution of acetone, followed by distilling off acetone from the combined water-acetone extract, the removal of impurities alcaloides character from acidified with sulfuric acid water balance single by treatment with ethylene dichloride, the isolation of alkaloids from alkalized solution 2-3-fold extraction with dichloromethane or ethyl acetate, the processing of alkaloids alcoholic solution of Hydrobromic acid and a single recrystallization of lappaconitine hydrobromide from aqueous alcohol ( EN 2123347, 1998 [5] ).

The disadvantages of this method include the duration of the production cycle (approximately 60 hours: 21-28 hours spent on the extraction of vegetable raw materials by 3-5-fold extraction, some time is spent on evaporation of the extractant, spend 4 extraction dichloroethane, spent about 24 hours for crystallization of the base lappaconitine and about 10 hours on Crist�lisalu lappaconitine hydrobromide), significant amounts of resources spent on the production cycle, the possibility of occurrence of side reactions of hydrolysis is not sufficiently high yield (average yield of the target product is about 0,595% based on the dry raw material source).

The object of the present invention is to simplify the technological process, reduce its duration and increase the yield of the target product pharmacopoeial purity.

This goal is achieved by the proposed method of obtaining lappaconitine hydrobromide, which is as follows.

The roots or grass of Northern monkshood (Aconitum septentrioale) or Aconite beloustogo (Aconitum leucostomum) is subjected to extraction with methylene chloride in the apparatus for continuous extraction, the resulting extract was directly subjected to flash chromatography.

As a sorbent for flash chromatography using alumina or silica gel. Thus adsorbed alkaloids eluted with an organic solvent, such as acetone or ethyl acetate. Received ethylacetate the solution of the base lappaconitine was evaporated, dissolved in acetone and the acetone solution of the base lappaconitine treated with a solution of Hydrobromic acid in acetone, the crude hydrobromide lapponica filtered, dried and a single recrystallization from ethanol poluchitekonomiyu of lapponian pharmacopoeial purity.

As a sorbent for flash chromatography is preferably used silica gel using as eluent ethyl acetate.

As a sorbent for flash chromatography preferably using aluminum oxide using as eluent acetone.

In an apparatus for continuous extraction using conventional apparatus for extraction in the system "solid - liquid", such as a Soxhlet apparatus, the percolators.

The proposed method using the continuous extraction of the source plant material, in addition, can significantly reduce the volume of solvent used. As an extraction using methylene chloride, which is a cheap low-boiling solvent. Extraction with methylene chloride boiling does not cause gumming, because its boiling point is at 42 º C.

It should be noted that the proposed method using the continuous extraction of the initial plant raw material with methylene chloride eliminates the acidification and subsequent alkalization of the extracts, that is, to eliminate the possibility of occurrence of side reactions of hydrolysis. For purification of the extract obtained was passed through a thin layer of sorbent (flash chromatography) than is achieved sorption amounts of alkaloids and more polar impurities� on the sorbent and the removal of low-polarity of colored impurities.

This goal is achieved also by a variant of the proposed method of obtaining lappaconitine hydrobromide carried out as follows:

The roots or grass of Northern monkshood (Aconitum septentrioale) or Aconite beloustogo (Aconitum leucostomum) is subjected to extraction with a polar organic solvent selected from the group comprising ethanol, acetone and aqueous solutions, with subsequent removal from the extract polar organic solvent, alkalization and extraction of the obtained residue with methylene chloride, followed by purification of the obtained extract flash chromatography.

As a sorbent for flash chromatography using alumina or silica gel. Thus adsorbed alkaloids eluted with an organic solvent, such as acetone or ethyl acetate. Received ethylacetate the solution of the base lappaconitine was evaporated, dissolved in acetone and the acetone solution of the base lappaconitine treated with a solution of Hydrobromic acid in acetone, the crude hydrobromide lapponica filtered, dried and a single recrystallization from ethanol hydrobromide lapponica pharmacopoeial purity

As a sorbent for flash chromatography is preferably used silica gel using as eluent ethyl acetate.

As a sorbent for flash x�omatography it is preferable to use aluminum oxide using as eluent acetone.

In this method, the removal of the polar organic solvent from the extract is carried out under reduced pressure, and then the distillation residue basified to pH 8-10 and it was extracted with methylene chloride. The resulting solution of the chloride methylene continue to use the same sequence for flash chromatography, as in the case of the proposed method using continuous extraction.

Extraction of plant raw materials using polar solvent in one of the embodiments of the present method is carried out in the apparatus for continuous extraction. This embodiment of the claimed method allows to obtain an additional reduction in the duration of the process.

In the proposed methods for cleaning floridametroguide extract using flash chromatography, passing the extract through a thin layer of sorbent, due to sorption amounts of alkaloids and more polar impurities on the sorbent, and the removal of low-polarity of colored impurities.

As a sorbent for flash chromatography using alumina or silica gel. Thus adsorbed alkaloids eluted with an organic solvent, for example ethyl acetate or acetone. Evaporation ethylacetate or acetone solution yields a crude base lappaconitine, the content of the target and�of kaleida which reaches 80%. Thus, used in the method eluent (ethyl acetate or acetone) is almost completely regenerated and can be reused for the next elution.

The resulting crude base lappaconitine was dissolved in a minimal amount of acetone, cooled and carefully acidified with 48% solution of Hydrobromic acid in acetone. The precipitate was lappaconitine hydrobromide (VFS) was filtered, washed with a minimal amount of cold acetone, methylene chloride and dried.

In the case of using aluminum oxide, for elution of the crude base lappaconitine used acetone. In this case, the solution obtained is the Foundation of lappaconitine is concentrated by evaporation of the solvent under vacuum and then spend the acidification, and lappaconitine hydrobromide allocate and clear quite similarly to the case of using as eluent ethyl acetate.

The purity of the suggested methods of preparation of lappaconitine hydrobromide is over 90%. Posttreatment to pharmacopoeial purity is achieved with a single recrystallization from ethanol.

After a single crystallization from ethanol, the purity of lappaconitine hydrobromide greater than 97%.

In the proposed method using a continuous extraction of vegetable raw materials during manufacturing process�and is about 30 hours, and in the case of extraction of plant raw material polar organic solvent is about 12 hours. The average yield based on starting the dry raw material in the proposed methods is 0.73% (which is 20% higher than the average yield according to a known method).

Thus, the intended objective, namely, to simplify the process, reduce its duration and increased yield of the target product pharmacopoeial purity is achieved suggested methods of obtaining lappaconitine hydrobromide (options), based on a fundamentally new approach to cleaning lappaconitine, which is based on the method of flash chromatography for the purification of the obtained alkaloid extract by passing it through a thin layer of sorbent, due to sorption amounts of alkaloids and more polar impurities on the sorbent, and the removal of low-polarity of colored impurities. This highly effective method has never been used to obtain lappaconitine pharmacopoeial purity. This method does not require the use of special equipment and devices, allows the use of much smaller quantities of sorbent and eluent, and, in addition, the process chromatography takes less time and also the method is easily scaled.

Thus, the proposed SPO�Oba represent a very flexible and effective approach to obtaining lappaconitine hydrobromide. Depending on requirements process requirements of various equipment, as well as a broad range of solvents. No matter which sequence greatly reduces the number of stages of the process provides a high yield and degree of purity of the VFS with only one recrystallization.

The invention is further illustrated by the following specific examples of the invention.

The results of the allocation of lappaconitine hydrobromide from vegetable raw materials in the conditions of the proposed methods are summarized in the Table.

EXAMPLES

Example 1

The use of continuous extraction (methylene chloride), sorbent - silica

In a Soxhlet apparatus for continuous extraction with 500 ml load 250 g, ground to a fine powder of dry plant material containing the lappaconitine (the roots of Aconitum beloustogo). The extraction is carried out with 250 ml of methylene chloride for 24 hours (18 hours extraction is achieved about 90% of the target substance). The resulting extract was passed by gravity through a layer of silica gel on the filter with a porous plate (D = 3 cm, H layer = 4 cm, weight of silica gel ~12 g). The rest of methylene chloride, fused by gravity, pumped to vacuum to dryness. Eluent (methylene chloride) yellow-brown, aderrasi� low-polar colored impurities cast, with further regeneration of methylene chloride on a rotary evaporator.

The filter is applied 25 ml of ethyl acetate and allowed to drain by gravity. This portion of the eluent swing, sending it to the regeneration of ethyl acetate.

Target lappaconitine eluted from the silica gel ~150 ml of ethyl acetate (control - TLC, Silufol, acetone, Rflappaconitine ~0,65), and the resulting solution lappaconitine evaporated on a rotary evaporator to dryness, yielding about 1.5 g of a yellow viscous, crystallizing on standing oil. According to HPLC the content of the target lappaconitine in this balance reaches 80%.

The residue was dissolved in 3 ml of acetone and cooled to -10°C. To the obtained solution under stirring dropwise from the dropping funnel was added a solution prepared in advance ~0.3 ml of 48% Hydrobromic acid in 2 ml of acetone (control - universal indicator paper, pH ~5). Triturated with a spatula precipitated white crystalline precipitate of lappaconitine hydrobromide, leave in the freezer at -18°C for 30 minutes and then filtered on a filter with a porous plate. The filter cake was washed with a minimal amount of cold acetone (3 ml) and then washed with methylene chloride (5 ml) and dried. Get about 1.5 g of lappaconitine hydrobromide, purity according to HPLC over 90%.

For the final cleaning of lappaconitine hydrobromide it crystallized from �of pirta, the rate of 2 ml alcohol on 850 mg of hydrobromide. Get about of 1.61 g of lappaconitine hydrobromide purity more than 97%.

The total yield of extraction of lapponica from plant material in the case of this approach is 0,645%.

Example 2

The use of continuous extraction (methylene chloride), sorbent alumina

In a Soxhlet apparatus for continuous extraction with 500 ml load 250 g, ground to a fine powder of dry plant material containing the lappaconitine (the roots of Aconite North). The extraction is carried out with 250 ml of methylene chloride for 24 hours (18 hours extraction is achieved about 90% of the target substance). The resulting extract was passed by gravity through a layer of aluminum oxide on the filter with a porous plate (D = 3 cm, H layer = 3 cm, weight of aluminum oxide ~15 g). The rest of methylene chloride, fused by gravity, pumped to vacuum to dryness. Eluent (methylene chloride) yellow-brown color, containing low-polarity colored impurities cast, with further regeneration of methylene chloride on a rotary evaporator.

Target lappaconitine eluted from the alumina with ~200 ml of acetone (control - TLC, Silufol, acetone, Rflappaconitine ~0,65). The resulting solution lappaconitine concentrated on a rotary evaporator to a volume of 3-4 ml. Obtained�th residue is cooled to -10°C with stirring dropwise from the dropping funnel was added a solution prepared in advance ~0.3 ml of 48% Hydrobromic acid in 2 ml of acetone (control - universal indicator paper, pH ~5). Triturated with a spatula precipitated white crystalline precipitate of lappaconitine hydrobromide, leave in the freezer at -18°C for 30 minutes and then filtered on a filter with a porous plate. The filter cake was washed with a minimal amount of cold acetone (3 ml) and then washed with methylene chloride (5 ml) and dried. Get about 1.3 g of lappaconitine hydrobromide, purity according to HPLC over 90%.

For the final cleaning of lappaconitine hydrobromide it crystallized from alcohol, at the rate of 2 ml alcohol on 850 mg of hydrobromide. Get about 1.25 g of lappaconitine hydrobromide purity more than 97%.

The total yield of extraction of lappaconitine from plant material in the case of this approach is 0.5%.

Example 3

The method is carried out analogously to example 2, but using in the vegetable raw materials are grass Aconite beloustogo. The conditions and results of the allocation of lappaconitine hydrobromide is presented in the Table below.

Example 4

The method is carried out analogously to example 2, but using in the vegetable raw materials of the herb Aconite North. The conditions and results of the allocation of lappaconitine hydrobromide is presented in the Table below.

Example 5

The use of continuous extraction (acetone), the sorbent is silica gel.

In the Soxhlet extraction d�I continuous extraction with 500 ml load 250 g, ground to a fine powder of dried plant material containing lappaconitine (the roots of Aconite North). The extraction is carried out with 250 ml of acetone for 4 hours. The resulting extract was evaporated to dryness on a rotary evaporator, the residue dissolved in 250 methylene chloride and passed by gravity through a layer of silica gel on the filter with a porous plate (D = 3 cm, H layer = 4 cm, weight of silica gel ~12 g). The rest of methylene chloride, fused by gravity, pumped to vacuum to dryness. Eluent (methylene chloride) yellow-brown color, containing low-polarity colored impurities, is cast away, with further regeneration of methylene chloride on a rotary evaporator.

The filter is applied 25 ml of ethyl acetate and allowed to drain by gravity. This portion of the eluent swing, sending it to the regeneration of ethyl acetate.

Target lappaconitine eluted from the silica gel ~150 ml of ethyl acetate (control - TLC, Silufol, acetone, Rflappaconitine ~0,65), and the resulting solution lappaconitine evaporated on a rotary evaporator to dryness, yielding about 2.5 g of a yellow viscous, crystallizing on standing oil. According to HPLC the content of the target lappaconitine in this balance reaches 80%.

The residue was dissolved in 4 ml of acetone and cooled to -10°C. To the obtained solution under stirring dropwise from the dropping funnel was added a solution prepared in advance ~0.3 ml of 48% Hydrobromic acid in 2 ml of AC�tone (control - universal indicator paper, pH ~5). Triturated with a spatula precipitated white crystalline precipitate of lappaconitine hydrobromide, leave in the freezer at -18°C for 30 minutes and then filtered on a filter with a porous plate. The filter cake was washed with a minimal amount of cold acetone (3 ml) and then washed with methylene chloride (5 ml), and dried. Get about 1.8 g of lappaconitine hydrobromide, purity according to HPLC over 90%.

For the final cleaning of lappaconitine hydrobromide it crystallized from alcohol, at the rate of 2 ml alcohol on 850 mg of hydrobromide. Get about 2.0 g of lappaconitine hydrobromide purity more than 97%.

The total yield of extraction of lapponica from plant material in the case of this approach is 0.8%.

Example 6

The use of conventional extraction (80% ethanol), the sorbent is silica gel.

In a round bottom flask of 1 l was charged with 100 g of milled to a fine powder of dry plant material containing the lappaconitine (the roots of Aconitum beloustogo). Add 900 ml of 80% aqueous ethanol and stirred for 2 hours. Decanted the solvent through a filter paper. To the residue in the flask, add 900 ml of 80% aqueous ethanol and stirred for 2 hours. The solvent is decanted through a paper filter, combined with the previous extract and ethanol is evaporated � vacuum (50 mbar). The residue was added 20 ml of 10% aqueous sodium carbonate solution (pH ~9) and the resulting solution was extracted with methylene chloride (3 times 50 ml). The combined extract was dried with anhydrous sodium sulfate and passed by gravity through a layer of silica gel on the filter with a porous plate (D = 3 cm, H layer = 3 cm, weight of silica gel ~9 g). The rest of methylene chloride, fused by gravity, pumped to vacuum to dryness. Eluent (methylene chloride) yellow-brown color, containing low-polarity colored impurities, is cast away, with further regeneration of methylene chloride on a rotary evaporator.

Target lappaconitine eluted from the silica gel ~150 ml of ethyl acetate (control - TLC, Silufol, acetone, Rflappaconitine ~0.65) and the resulting solution is the Foundation of lappaconitine evaporated on a rotary evaporator to dryness, yielding about 1.3 g of a yellow viscous, crystallizing on standing oil.

The residue was dissolved in 3 ml of acetone and cooled to -10°C. To the obtained solution under stirring dropwise from the dropping funnel was added a solution prepared in advance ~0.3 ml of 48% Hydrobromic acid in 2 ml of acetone (control - universal indicator paper, pH ~5). Triturated with a spatula precipitated white crystalline precipitate of lappaconitine hydrobromide, leave in the freezer at-18ºC for 30 minutes and then filtered on a filter with a porous �plastinkoi. The filter cake was washed with a minimal amount of cold acetone (3 ml) and then washed with methylene chloride (5 ml) and dried. Get about 1.1 g of lappaconitine hydrobromide, purity according to HPLC over 90%.

For the final cleaning of lappaconitine hydrobromide it crystallized from alcohol, at the rate of 2 ml alcohol on 850 mg of hydrobromide. Get about 0.95 g lappaconitine hydrobromide purity more than 97%.

The yield extract lappaconitine from plant material in the case of this approach is 0.95%.

Example 7

The use of conventional extraction (80% ethanol), the sorbent is alumina.

In a round bottom flask of 1 l was charged with 100 g of milled to a fine powder of dry plant material containing the lappaconitine (the roots of Aconite North). Add 900 ml of 80% aqueous ethanol and stirred for 2 hours. Decanted the solvent through a filter paper. To the residue in the flask, add 900 ml of 80% aqueous ethanol and stirred for 2 hours. The solvent is decanted through a paper filter, combined with the previous extract and ethanol is evaporated under vacuum (50 mbar). The residue was added 20 ml of 10% aqueous sodium carbonate solution (pH ~9) and the resulting solution was extracted with methylene chloride (3 times 50 ml). The combined extract was dried with anhydrous sodium sulfate and passed� by gravity through a layer of aluminum oxide on the filter with a porous plate (D=3 cm, N. layer = 3 cm, weight of aluminum oxide ~15 g). The rest of methylene chloride, fused by gravity, pumped to vacuum to dryness. Eluent (methylene chloride) yellow-brown color, containing low-polarity colored impurities, is cast away, with further regeneration of methylene chloride on a rotary evaporator.

Target lappaconitine eluted with aluminum oxide ~200 ml of acetone (control - TLC, Silufol, acetone, Rflappaconitine ~0,65), and the resulting solution is the Foundation of lappaconitine evaporated on a rotary evaporator to a volume of 3-4 ml. Obtained residue was cooled to -10°C with stirring dropwise from the dropping funnel was added a solution prepared in advance ~0.3 ml of 48% Hydrobromic acid in 2 ml of acetone (control universal indicator paper, pH ~5). Triturated with a spatula precipitated white crystalline precipitate of lappaconitine hydrobromide, leave in the freezer at -18°C for 30 minutes and then filtered on a filter with a porous plate. The filter cake was washed with a minimal amount of cold acetone (3 ml) and then washed with methylene chloride (5 ml), and dried. Get about 0.9 g lappaconitine hydrobromide, purity according to HPLC over 90%.

For the final cleaning of lappaconitine hydrobromide it crystallized from alcohol, at the rate of 2 ml alcohol on 850 mg of hydrobromide. Get about 0.75 g of the hydrobromide La�of aconitine purity more than 97%.

The total yield of extraction of lappaconitine from plant material in the case of this approach is 0.75%.

Example 8

The use of conventional extraction (80% aqueous acetone), the sorbent is silica gel.

In a round bottom flask of 1 l was loaded with 100 g of milled to a fine powder of dry plant material containing the lappaconitine (the roots of Aconitum beloustogo). Add 900 ml of 80% aqueous acetone and stirred for 2 hours. Decanted the solvent through a filter paper. To the residue in the flask, add 900 ml of 80% aqueous acetone and stirred for another 2 hours. The solvent is decanted through a paper filter and is combined with the previous extract. Evaporated the acetone under vacuum (60 mbar). The residue was added 20 ml of 10% aqueous sodium carbonate solution (pH ~9) and the resulting solution was extracted with methylene chloride (3 times 50 ml). The combined extract was dried with anhydrous sodium sulfate and passed by gravity through a layer of silica gel on the filter with a porous plate (D = 3 cm, H layer = 3 cm, weight of silica gel ~9 g). The rest of methylene chloride, fused by gravity, pumped to vacuum to dryness. Eluent (methylene chloride) yellow-brown color, containing low-polarity colored impurities, is cast away, with further regeneration of methylene chloride on a rotary evaporator.

Alavalapati eluted from the silica gel ~150 ml of ethyl acetate (control - TLC, Silufol, acetone, Rflappaconitine ~0.65) and the resulting solution is the Foundation of lappaconitine evaporated on a rotary evaporator to dryness, yielding about 1.2 g of a yellow viscous, crystallizing on standing oil.

The residue was dissolved in 3 ml of acetone and cooled to -10°C. To the obtained solution under stirring dropwise from the dropping funnel was added a solution prepared in advance ~0.3 ml of 48% Hydrobromic acid in 2 ml of acetone (control-universal indicator paper, pH ~5). Triturated with a spatula precipitated white crystalline precipitate of lappaconitine hydrobromide, leave in the freezer at -18°C for 30 minutes and then filtered on a filter with a porous plate. The filter cake was washed with a minimal amount of cold acetone (3 ml) and then washed with methylene chloride (5 ml), and dried. Get about 1.0 g of lappaconitine hydrobromide, purity according to HPLC over 90%.

For the final cleaning of lappaconitine hydrobromide it crystallized from alcohol, at the rate of 2 ml alcohol on 850 mg of hydrobromide. Get about 0.85 g lappaconitine hydrobromide purity more than 97%.

The total yield of extraction of lappaconitine from raw materials in the case of using this approach is about 0.85%.

Example 9

The use of conventional extraction (80% aqueous acetone), the sorbent is alumina.

In a round bottom flask of 1 l loaded�up with 100 g of milled to a fine powder of dried plant material containing lappaconitine (the roots of Aconite North). Add 900 ml of 80% aqueous acetone and stirred for 2 hours. Decanted the solvent through a filter paper. To the residue in the flask, add 900 ml of 80% aqueous acetone and stirred for another 2 hours. The solvent is decanted through a paper filter and is combined with the previous extract. Evaporated the acetone under vacuum (60 mbar). The residue was added 20 ml of 10% aqueous sodium carbonate solution (pH ~9) and the resulting solution was extracted with methylene chloride (3 times 50 ml). The combined extract was dried with anhydrous sodium sulfate and passed by gravity through a layer of aluminum oxide on the filter with a porous plate (D = 3 cm, H layer = 3 cm, weight of aluminum oxide ~15 g). The rest of methylene chloride, fused by gravity, pumped to vacuum to dryness. Eluent (methylene chloride) yellow-brown color, containing low-polarity colored impurities, is cast away, with further regeneration of methylene chloride on a rotary evaporator.

Target lappaconitine eluted with aluminum oxide ~200 ml of acetone (control - TLC, Silufol, acetone, Rflappaconitine ~0.65) and the resulting solution is the Foundation of lappaconitine evaporated on a rotary evaporator to a volume of 3-4 ml. Obtained residue was cooled to -10°C and with stirring, dropwise from the dropping funnel, add cooked �the Aranea a solution of ~0.3 ml of 48% Hydrobromic acid in 2 ml of acetone (control - universal indicator paper, pH ~5). Triturated with a spatula precipitated white crystalline precipitate of lappaconitine hydrobromide, leave in the freezer at -18°C for 30 minutes and then filtered on a filter with a porous plate. The filter cake was washed with a minimal amount of cold acetone (3 ml) and then washed with methylene chloride (5 ml) and dried. Get about 0.9 g lappaconitine hydrobromide, purity according to HPLC over 90%.

For the final cleaning of lappaconitine hydrobromide it crystallized from alcohol, at the rate of 2 ml alcohol on 850 mg of hydrobromide. Get about 0.7 g of lappaconitine hydrobromide purity more than 97%.

The total yield of extraction of lappaconitine from raw material when using this approach amounts to 0.7%.

Table
The conditions and results of the allocation of lappaconitine hydrobromide from plant material
№ p/pThe raw material mass gThe type of extraction, the extractantThe output of the Pharmacopoeia of lappaconitine hydrobromide (VFS)Used vegetable raw materials
g% by weight of suhag� raw materials
1250continuous, methylene chlorideto 1.610,645The roots of Aconitum beloustogo
2250continuous, methylene chloride1,250,5The roots of Aconite North
3250continuous, methylene chloride0,8750,350Herb Aconite beloustogo
4250continuous, methylene chloride0,750,300Herb Aconite North
5250continuous,
acetone
20,8The roots of Aconite North
6100usual,
80% ethanol
0,95 0,95The roots of Aconitum beloustogo
7100usual,
80% ethanol
0,750,75The roots of Aconite North
8100common, 80% aqueous acetone0,850,85The roots of Aconitum beloustogo
9100common, 80% aqueous acetone0,70,7The roots of Aconite North

1. A method of producing lappaconitine hydrobromide by extraction of roots or herbs of Northern monkshood (Aconitum septentrioale) or Aconite beloustogo (Aconitum leucostomum) organic solvent, purification of the extract, the processing of lappaconitine with a solution of Hydrobromic acid and crystallization of the target product, characterized in that the extraction ofspecified vegetable raw materials is carried out with methylene chloride in the apparatus for continuous extraction with subsequent purification of the obtained extract flash chromatography.

2. A method according to claim 1, where the sorbent for flash chromatography�raffia using silica gel, but as eluent using ethyl acetate.

3. A method according to claim 1, where the sorbent to flash chromatography using aluminum oxide, and the eluent used is acetone.

4. A method of producing lappaconitine hydrobromide by extraction of roots or herbs of Northern monkshood (Aconitum septentrioale) or Aconite beloustogo (Aconitum leucostomum) organic solvent, purification of the extract, the processing of lappaconitine with a solution of Hydrobromic acid and crystallization of the target product, characterized in that the extraction ofspecified vegetable raw materials is carried out in polar organic solvent selected from the group comprising ethanol, acetone and aqueous solutions, with subsequent removal from the extract polar organic solvent, alkalization and extraction of the obtained residue with methylene chloride, followed by purification of the obtained extract flash chromatography.

5. A method according to claim 4, where the sorbent to flash chromatography using silica gel and as the eluent using ethyl acetate.

6. A method according to claim 4, where as a sorbent for flash chromatography using aluminum oxide, and the eluent used is acetone.

7. A method according to claim 4, wherein the extraction ofcarried out in an apparatus for continuous extraction.



 

Same patents:

FIELD: food industry.

SUBSTANCE: method for production of Siberian cedar seeds liqueur (with hepatoprotective, antioxidant, antihypoxic, hypolipidemic effect) by way of maceration with ethyl alcohol usage; whole Siberian cedar seeds are loaded into the reactor, poured with 70% ethyl alcohol water solution; extraction is performed under preset conditions. The medicinal preparation with hepatoprotective, antioxidant, antihypoxic, hypolipidemic effect contains Siberian cedar seeds liqueur. Usage of the medicinal preparation as a hepatoprotective remedy.

EFFECT: liqueur has pronounced hepatoprotective, antioxidant, antihypoxic and hypolipidemic effect.

6 cl, 3 dwg, 8 tbl

FIELD: medicine.

SUBSTANCE: method for producing a pigment complex of bisnaphthazarin for preventing inflammatory diseases, involving demineralising commercial sea urchins' crusts and needles in an organic acid solution, separating organic acid salts and protein, applying pigment solution on a chromatographic column, washing the column with diluted mineral acid and distilled water, eluting the pigment complex, combining fractions containing the pigments, removing ethanol, lyophilising concentrate in the certain environment. The complex of pigments bisnaphthazarins for preventing inflammatory diseases.

EFFECT: complex of pigments prepared by the above method is effective for preventing the inflammatory diseases.

3 cl, 2 dwg, 2 tbl, 4 ex

Vibration extractor // 2545300

FIELD: machine building.

SUBSTANCE: for extraction (leaching) of the substances extracted from the plant materials in food, chemical-pharmaceutical and other industries, for output increasing of the substances extracted from the extractable plant materials and for increasing of their concentration in the ready extraction it is suggested to provide the extractor with the extractant recirculation circuit containing devices for solid phase separation, pump, discharge tank, flowmeter, shutdown valves system, and in the extractor bottom part additionally union will be installed for continuous liquid phase supply.

EFFECT: wider possibility of return in the vessel of part of extraction after solid phase separation in specified ratio with fresh extractant, thus improving conditions of mass exchange due to decreasing of the surface tension of the liquid phase.

1 dwg

FIELD: medicine.

SUBSTANCE: method for preparing an agent possessing anti-inflammatory, diuretic and antioxidant activity, involving milling Spiraea salicifolia shoots representing a mixture of leaves, blossom and shoots, extracting them three times by gradual maceration, mixing in infusing, filtering, condensing, separating, drying in the certain environment.

EFFECT: agent shoes the pronounced anti-inflammatory, diuretic and antioxidant activity.

2 dwg, 12 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical and consumer industry in preparing dry herbal extracts used for later colouration of textile fabrics with this extract. A method for preparing the dry herbal extract of St. John's wort involves grinding herbal raw material, extracting in water, filtering, boiling down in a rotary evaporator and drying the residual product to constant weight in a drying chamber in the certain environment.

EFFECT: method described above enables preparing the extract with dry colorant weight 25% of dry herbal raw material weight.

FIELD: chemistry.

SUBSTANCE: method of obtaining sanguinarine and chelerythrine sulphates includes extraction of milled overground part of macleaya microcarpa and/or macleaya cordata with water aliphatic alcohol, removal of water aliphatic alcohol in vacuum, alkalinisation of water distillation residue with hydrophobic solvent, processing organic phase with sulphuric acid, filtration, washing and drying of target product, with extraction of milled raw material being carried out with water aliphatic alcohol in presence of methanesulphoacid, and solution of alkaloid bases in hydrophobic organic solvent is additionally filtered through layer of hydrophobic solvent, and target product is subjected to boiling in acetone.

EFFECT: method makes it possible to increase quality of finished product, simplify technology of production of sanguinarine and chelerythrine sulphates, and reduce the process duration.

8 cl, 14 ex

FIELD: chemistry.

SUBSTANCE: method of bee pollen processing consists in the fact, that bee pollen is extracted with CO2 by pumping CO2, obtained fat extract is separated, remaining oil-seed meal is subjected to enzymatic hydrolysis in presence of enzyme Distizym Protacide Extra, obtained fermentolisate is separated into solid and liquid phases, solid phase is dried, liquid phase is filtered, preservative potassium sorbate and sodium benzoate are added into filtered liquid fraction under specified conditions.

EFFECT: method makes it possible to efficiently process bee pollen with obtaining fractions, characterised by microbiological purity, high degree of bioavailability, and a long storage term.

4 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to versions of composition for heat transmission. One of composition versions contains (i) from about 20 to about 90 wt % of R-1234yf; (ii) from about 10 to about 60 wt % of R-134a and (iii) from about 1 to about 20 wt % of R-32. Invention also relates to a number of versions of composition application.

EFFECT: composition has lower value of global warming potential and at the same time is characterised by productivity and energy efficiency.

56 cl, 7 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, particularly to a method for recovering pinosylvin and methylpinosylvin from common pine (Pinus sylvestris) wood. The method for recovering pinosylvin and methylpinosylvin consists in extracting ground knots of common pine (Pinus sylvestris) wood, common pine (Pinus sylvestris) in isopropyl alcohol mixed with water, separating the extract and removing the solvent.

EFFECT: method enables a higher yield of pinosylvin and methylpinosylvin from common pine (Pinus sylvestris) wood.

3 cl, 5 ex, 2 dwg

FIELD: medicine.

SUBSTANCE: agent possessing the anti-inflammatory, antipyretic and antimicrobial action representing a dry extract of drug hedge hyssop leaves and blossom by grinding them, extracting in 96% alcohol on a water bath to a boil, and boiling, evaporating, diluting the evaporated residue by distilled water first, adding chloroform then, cooling to a room temperature and centrifuging, separating a water fraction and drying it in the certain environment.

EFFECT: agent possesses the pronounced anti-inflammatory, antipyretic and antimicrobial action.

5 dwg, 5 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: agent for treating diabetes mellitus involving dry aqueous extract of Geranium Dieisianium Knuth and dry aqueous extract of Uncaria tomentosa (Willd) D.C. bark enclosed into gelatine capsules. A method of treating diabetes mellitus provides prescribing the above agent in a daily dose of 180-360 mg after a meal. The above agent enables treating diabetes mellitus effectively by reducing dosages of blood glucose lowering drugs.

EFFECT: agent causes the positive effect on carbohydrate metabolism and possesses non-specific immunomodulatory action.

2 cl, 5 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to chemical-pharmaceutical industry and represents composition for oral cavity care, containing: a) at least one compound of formula M1-A-M2-B-M3, where: M1 and M3 represent Ti or titanium (Ti) oxide; A and B represent C2-C6 dibasic acid; and b) at least one orally acceptable solvent; where composition contains less than 5% of water.

EFFECT: invention provides creation of composition for treatment of teeth hypersensitivity, which effectively seals open dentin canals.

7 cl, 8 tbl, 12 dwg, 1 ex

FIELD: biotechnology.

SUBSTANCE: method comprises dissolving the mixture of birch bark triterpenoids in tetrahydrofuran to obtain the solution with a concentration of 5-10 g/l. Oleic acid is dissolved in an amount of 5-10% by weight of the birch bark triterpenoids. The sterilising filtration of the mixture is carried out. 25-fold excess of 0.01 M tris buffer is added, pH 9.0±0.2, when stirring. Sonication is carried out for 5-10 min. The organic solvent is removed using ultrafiltration on hollow membranes with exclusion threshold of 300 kDa at a rate of 1.0-1.2 L/min at a pressure of 0.6-0.8 atm. Cryoprotectant is added from the group of substances: mannitol, maltose, trehalose, mannose, sorbite, sucrose. The resulting concentrated mixture is frozen.

EFFECT: invention enhances the immunogenic activity of viral vaccines and provides their stability while storage.

2 cl, 2 dwg, 6 tbl, 7 ex

FIELD: medicine.

SUBSTANCE: group of inventions refers to an anti-ageing product. Moringa sp. whole seed extract for an anti-ageing effect containing oil and polyphenols representing an extract prepared of a moderately polar solvent. Using Moringa sp. whole seed extract as an active anti-ageing ingredient. Using Moringa sp. whole seed extract for enhancing and recovering a skin barrier function. A cosmetic and/or dermatologic anti-ageing composition. A cosmetic method for the anti-ageing effect in the individuals with mature skin, involving using topical or oral administration of Moringa sp. whole seed extract. A method for preparing Moringa sp. whole seed extract.

EFFECT: extract is effective for the anti-ageing effect.

12 cl, 1 tbl, 5 ex, 1 dwg

FIELD: food industry.

SUBSTANCE: method for production of Siberian cedar seeds liqueur (with hepatoprotective, antioxidant, antihypoxic, hypolipidemic effect) by way of maceration with ethyl alcohol usage; whole Siberian cedar seeds are loaded into the reactor, poured with 70% ethyl alcohol water solution; extraction is performed under preset conditions. The medicinal preparation with hepatoprotective, antioxidant, antihypoxic, hypolipidemic effect contains Siberian cedar seeds liqueur. Usage of the medicinal preparation as a hepatoprotective remedy.

EFFECT: liqueur has pronounced hepatoprotective, antioxidant, antihypoxic and hypolipidemic effect.

6 cl, 3 dwg, 8 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to cosmetology and dermatology and represents a skin care composition applicable for local skin application, wherein the above composition contains salicylic acid or its salt in a combination with glycyrrhizic acid, or its salt or its derivative, cetylhydroxyproline palmitamide, lactic acid or its salt, bisabolol and niacinamide.

EFFECT: invention provides extending the range of effective skin care agents.

41 cl, 11 ex, 11 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions concerns oral care compositions effective for treating dental hypersensitivity, and a method of treating using them. The composition contains a compound of formula I: M1-A-M2-B-M1; wherein M1 and M3 represent potassium (K); M2 represents Ti or titanium (Ti) oxide; A and B independently represent C2-C6 dibasic acid; and at least one orally acceptable solvent. The oral care composition has pH falling within the range of 2.0 to 7.0. There are also presented a version of the composition, which contains at least one additional desensitising agent, and a method of treating sensitive teeth with the use of this composition.

EFFECT: using the group of inventions provides the effective dental desensitisation by creating a protective barrier on the tooth surface and/or sealing the effective dental tubules effectively.

11 cl, 8 tbl, 12 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to the field of organic chemistry, namely to novel pyridine derivatives of the general formula

and to their pharmaceutically acceptable salts, where R1 stands for (C1-6) alkyloxy, CN or halogen, R2 stands for a hydrogen atom, R3 stands for a hydrogen atom or (C1-6) alkyl, R4, R5, R6, R7 are similar or different and stand for a hydrogen atom or halogen. The invention also relates to the cosmetic application of the formula (I) compound.

EFFECT: novel pyridine derivatives, useful in the treatment of diseases associated with a receptor of androgens, are obtained.

9 cl, 1 tbl, 16 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to health-promoting compositions and methods for preparing them. A method for preparing the composition of non-living lactic acid bacilli, possessing an ability of specific binding to Streptococcus mutans, involves the following stages: heating a cell suspension of lactic acid bacillus or a mixture of lactic acid bacilli possessing an ability of specific binding to Streptococcus mutans from an initial temperature of less than 40°C to a pasteurisation temperature of 75 to 85°C with a temperature variation within the range of 0.5 to 2°C/min, keeping the heated suspension at a pasteurisation temperature of 20 to 40 minutes and cooling the suspension to a final temperature of less than 40°C within the range of 0.5 to 2°C/min. The specific binding the cell suspension of the lactic acid bacillus or the mixture of lactic acid bacilli to Streptococcus mutans is stable to heat treatment and/or resistant to proteases and/or calcium-dependent and/or is observed within the range of pH values falling within the range of 4.5 and 8.5, and/or in the saliva environment.

EFFECT: invention enables producing the agent preventing or delaying the caries lesion formation.

9 cl, 4 ex

FIELD: medicine.

SUBSTANCE: 0.5% dihydroquercetin is instilled into the rectum of a patient with temporary colostomy until he/she starts feeling intestinal inflation. The procedure is performed twice a day, daily up until the restorative surgery.

EFFECT: method reduces a rate and a degree of colitis manifestations by the local antioxidant, anti-inflammatory effect, unfolding the excluded colon.

FIELD: medicine, oncology, amino acids.

SUBSTANCE: invention relates, in particular, to the development of an antitumor preparation based on natural substances. Invention relates to an amino acid preparation comprising at least one modified essential amino acid obtained by treatment of amino acid by ultraviolet radiation (UV) at wavelength 250-350 nm for 12-80 h at temperature 15-30oC or with ozone at temperature 15-25oC. The modified amino acid has no toxicity for health cells. Also, invention relates to a method for preparing such preparation. Invention provides the development of an antitumor preparation based on modified amino acids and expanded assortment of antitumor preparations being without cytotoxicity for normal cells.

EFFECT: valuable medicinal antitumor properties of preparation.

8 cl, 4 tbl, 2 dwg, 4 ex

Up!