RO127000B1 - Bioactive complex of triterpene acids, process of preparation and medicinal products - Google Patents

Abstract

The invention relates to a bioactive complex of triterpene acids, used as pharmacologically bioactive component in human and veterinary therapy and also to a process for preparing the same. The claimed complex is based on Salvia species herba, having crystalline structure, enhanced solubility and an acid content of 100%, consisting of at least 80% ursolic acid and ut most 20% of a mixture consisting of one or more of the acids: oleanoic, hydroxyursolic, hydroxyoleanolic, betulinic and dihydrobetulinic as free acids or as Na, K, or NHsalts. The claimed process consists in the extraction, by dynamic maceration of the dry and powdered Salvia species herba plant with 5...15% structured water with 85...95% acetone or 85...95% of 95% ethyl alcohol, for 8...24 h, at a temeparture of 15...80°C, followed by processing and concentrating the extract and possibly converting the acid mixture from the extract in Na, K or NHsalts.

Description

The invention relates to a bioactive complex of triterpenic acids, as such or as Na, K or NH ₄ salts, to the process of obtaining Salvia species from the plant, and to medicinal products for human and veterinary use, made on the basis thereof.

The bioactive complex, made according to the invention, has a triterpenic acid content of 100%, being characterized physico-chemical and pharmacotoxicological. The evaluation of the specific activity and the safety in administration allows its use as a basic bioactive component of some medicinal products for internal or external use, in human and veterinary therapeutics.

Based on the recently published pharmacological studies, regarding triterpenic acids (especially ursolic and oleanolic), they are known, although isomers, to exhibit some structural differences that cause a differentiation of the pharmacological effects which, although largely similar, are not identical. . Therefore, especially when considering the therapeutic use of a mixture of triterpenic acids, it is very important and strictly necessary to know their proportions precisely, because a variability in the content of the mixture can cause a variability of the pharmacological activity and the bioavailability of the product. [1], [2], [3], [4], [5],

Also, pharmacological studies carried out on different types of medicinal substances have shown the direct relationship between the physical-chemical properties of the active principles and the therapeutic activity, depending on the rate of release of the active substance from the pharmaceutical form, the intensity and the duration of the effects. The physico-chemical properties of substances influence their solubility in biological environments. Knowing this fact allows predicting the behavior of drug substances from the point of view of bioavailability, in different regions of the gastrointestinal tract. The information provided by bioavailability studies may lead to changes in the formulation of the finished product and its manufacturing process, as well as the extension of therapeutic indications. A main factor that can alter the bioavailability of the drug is the design of the active principle, represented by the crystallization form. By modifying the technological steps to make the substances of interest, their crystalline structure can be obtained by directing, in order to improve their solubility and bioavailability. Triterpenic acids being hydrophobic substances, present major problems in their solubilization. According to "The Merck Index - An Encyclopedia of Chemicals, Drugs, and Biologicals", - O'Neil Ed., Maryadele. J. - 2006, p. 1699, the solubility of ursolic acid in ethyl alcohol is 1: 178 (m: m).

Analyzing the patented procedures for obtaining pure or mixed triterpenic acids, it is found that they are presented mostly at the laboratory level and do not present the complex physico-chemical characterization for the obtained product (analytical methods for determining the purity, structure). crystalline or amorphous, diffraction studies, IR, etc.). It is only in recent years that patents for the production of ursolic and oleanolic acid based bioproducts in the form of low purity concentrates (30 ... 65%) have been published, evaluated analytically by HPLC or GC and intended use as nutritional supplements.

Overall evaluation of patents of invention and recent patent applications, which claim processes for obtaining triterpenic acids (especially ursolic and oleanolic), in pure state or in mixture, as such or as salts, with inorganic or organic bases , leads to their classification according to the following criteria:

- the sources of vegetable raw materials:

- leaves and grass, harvested from plants of spontaneous flora or crops, whose processing leads to obtaining purified products, with therapeutic applications;

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- fruit wastes, resulting as waste in the food industry, through the processing of which 1 are obtained concentrates with a low content of triterpenic acids (30 ... 65%), usable for obtaining nutritional supplements; 3

- the finished product that constitutes the main objective of the patented process:

- pure triterpenic acids, as such or as salts, 5

- a mixture of triterpenic acids, triglycerides, sugars, etc. obtained mainly from fruit wastes.7 The patents of the invention and recent patent applications are presented below, which present processes for obtaining triterpenic acids (especially ursolic and / or oleanolic). 9

- Patent GR 10011738, entitled "Method of separation of the ursolic and oleanolic acids from the leaves of Olea europaea is a process at laboratory level, which 11 envisages the extraction of the dried leaves of Olea europaea with ethyl alcohol 95 ° C, at ambient temperature, for 15 days, the precipitation of triterpenic acids from the alcoholic extract by diluting it with water, separating and drying the sediment, followed by its purification by successive extractions with petroleum ether. There is presented a method of analysis by chromatography in 15 thin layers, by means of which the sediment purified with petroleum ether, as constituted of ursolic acid, is qualitatively evaluated, and the extracts of petroleum ether, concentrated in sico, contain 17 oleanolic acid.

There is no mention of the quantitative evaluation of the ursolic and oleanolic acids thus separated, 19 in order to be able to assess the purity of the substances and the efficiency of the separation process.

- Patent US 6740778, entitled "Method of preparation of olea-21olic acid and / or olive acid", presents a method of preparing these acids as such or in the form of physiologically acceptable salts, using as the starting material the Olea 23 plant europaea from which the material left after the extraction of the olive oil is processed. As extraction solvents, hydrophilic organic solvents (alcohols, acetone) or lipophiles 25 (hexane, chloroform, ethyl acetate) are used, and the resulting extracts containing oleanolic acid or olive oil are purified by liquid-liquid extraction or chromatography on silica gel column. 27

It is mentioned that the triterpenic acids thus obtained have a purity of 85 ... 100%. and the respective salts have a purity of at least 90%. 29

Although the method leads to obtaining high purity substances, it has the disadvantage of technological flows that are difficult to apply on an industrial scale, both through the purification methods 31 envisaged and through the multitude of organic solvents used.

- US Patent 18186346, entitled "Method of obtaining 33 ursolic acid", presents a process for obtaining ursolic acid from fruit wastes of Oxycoccus quadripetelus G, by extraction with isopropyl alcohol, at ambient temperature, 35 for 24 hours, concentration of the alcoholic extract under vacuum to the residue, which is purified by extraction with petroleum ether and successive transformations into Na salt and purified ursolic acid. 37

An increase in the processing efficiency and the quantity of the obtained substance is mentioned, without specifying its purity. 39

- Patent RU 2108107, entitled: "Method for preparing a biologically active total of salts of triterpenic acids", presents a process for obtaining 41 triterpenic acids of species of white fir (Abiessp.), In the form of a total of salts of Na of triterpenic acids, which provides for the extraction of vegetable raw material with a mixture of 43 petroleum ether - ethyl acetate, the concentration of extractive solutions in the residue, which is dissolved in ethyl acetate, and the organic solution is alkalized with 2% NaOH solution at pH = 9, after which the alkaline aqueous extract 45, with pH = 7 ... 8 is concentrated in vacuo, until a dry powder extract is obtained, which constitutes the total of triterpenic acids, as Na salt, with a content of 47 38 ... 45%.

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The chemical structure and the name of the triterpenic acids obtained according to the patented procedure are not specified, but from the literature data, it is known that the species of conifers contain triterpenic compounds with chemical structures other than those of ursolic and oleanolic acids.

- Patent RU 2108803, entitled: "Method for the preparation of a total of biologically active triterpenic acids", presents a process for obtaining a total of triterpenic acids from the needles of white fir, by extraction with methyl-butyl ether, treatment of the extract organic with alkaline agent (2% aqueous NaOH solution), separation of the aqueous alkaline solution, acidification with 10% HCl solution, at pH = 2 and isolation of the finished product from the acid solution by extraction with methyl-butyl ether.

As in the previous patent, the name and chemical structure of the triterpenic acids thus isolated are not specified, mentioning only the amount of the resulting substance, without indicating its purity.

- Patent RU 2151139, entitled: "Method of obtaining a total of biologically active triterpenic acids", presents a process for extracting triterpenic acids from fresh needles or white fir bark, previously treated with steam, then extracted with solvents organic (methyl-butyl ether; mixture: benzene-ethyl acetate; mixture: petroleum-ethyl acetate ether), followed by the separation of total triterpenic acids by successively treating the organic extract with basic aqueous and acidic solutions.

The quantity of product obtained shall be mentioned, without specifying the chemical name of the triterpenic acids and the purity of the product.

In addition, all the 3 patents from the Russian Federation, presented, although they refer to obtaining a total of biologically active triterpenic acids, do not present any kind of pharmacological tests that show the biological activity of the obtained products on the basis of which it is possible to evaluate the possibilities of application in therapeutics.

A group of patents belonging to the company UNILEVER presents different processes for the preparation of concentrates from fruit wastes, which contain a mixture of ursolic, oleanolic acids and other components, being intended for use as nutritional supplements.

- Patent EP 1161879 and US patent application 2002037882 present 2 laboratory and pilot procedures - which provide for the extraction of dried apples with acetone at reflux (5O ... 58 ° C), the concentration of the extractive solution at the residue and its crystallization from 50/50 acetone-water mixture to 95/5 (laboratory process) or hexane (pilot process).

A mixture of ursolic and oleanolic acid is obtained in varying proportions, which also contains other polar and apolar components specific to apple shells. There is also a process for conditioning the triterpenic mixture by association with mono-, di-, triglycerides and oils containing unsaturated fatty acids, for use as nutritional supplements.

- Patent EP 1250852 and US patent application 2003049365 present a laboratory procedure for obtaining triterpenic acid concentrates (ursolic and oleanolic), as such or in the form of salts, which provides for the processing of apple shells previously treated with aqueous solutions acids (H ₃ PO ₄ ) at pH = 2 and with aqueous alkaline solutions (Na ₂ CO ₃ ) at pH = 8-12, followed by extraction with acetone at Soxhlet and concentration of extractive solution at sicc residue, which constitutes the ursolic acid concentrate and oleanolic with a content of 30 ... 65% in the ratio 4: 6, together with other components: sugars, glycerides and other triterpenic compounds.

The use of concentrates thus obtained in the preparation of food supplements in the form of capsules or other forms of conditioning is mentioned.

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- Patent CN 1358733 presents a process for the extraction of 1 ursolic acid from the leaves of the Ligustrum lucidum plant, which includes the following phases: extraction of the leaves in counter current with ethyl alcohol at reflux, the concentration of the extract at the residue, which is washed with water and dried. , and the dry extract is dissolved in ethyl alcohol and discolored, then concentrated and diluted with water, resulting in a precipitate; adjust the pH of the suspension 5 to 2 ... 2.5, then the precipitate is separated, washed with neutral lapH water, and dried under vacuum.

It is mentioned to obtain a crude extract, having as main component 7 ursolic acid, without indicating the quantitative evaluation and the purity of the product, as well as the other compounds existing next to the ursolic acid. 9

- CN 1508149 presents a method of extraction of total triterpenic acids from // ex kudingcha with ethyl alcohol as solvent by a process of reflux distillation or extraction with 11 microwaves, followed by a series of steps of separation by ultrafiltration, nanofiltration, macroporous resin chromatography and purification by decolorization with activated carbon and separation using ether or an organic acid, finally obtaining a total triterpenic acid concentration> 95%. 15

CN 1453289 presents a process of extraction and separation of ursolic acid from the paulownia plant leaves by extraction at reflux with methanol, filtration, precipitation, washing with 17 tons and 3 times recrystallization with ethanol or anhydrous methanol.

- WO 2005075614 (A1) shows the extraction of olive leaves with hexane, ethyl alcohol, 19 followed by the operations of concentration, separation on an extraction column in countercurrent supercritical carbon dioxide. 21

- CN 1557831 presents a process of extraction of ursolic acid from rosemary by Soxhlet extraction, concentration, dissolution in ethanol, discoloration with coal, crystallization, filtration 23 after 3 ... 6 hours obtaining crystals of ursolic acid of about 90% purity. .

- CN 101318985 reports a method for obtaining ursolic acid from the leaves of the plant 25 loquat with 95% ethyl alcohol as solvent by ultrasonication, followed by filtration through the nanofiltration micromembrane, recovery of ethanol and obtaining ursolic acid by mixing with honey, 27 in the form of a cream with a ursolic acid content> 75%.

- CN 101538271 presents a method for simultaneous extraction of picrinin and ursolic acid 29 by extraction from aqueous aqueous solution of the leaves of the said plant subjected to ultrasound, the extract obtained is volatilized, the concentrate is alkalized and subjected to 31 extraction with chloroform, of which, by separation, picrinin and ursolic acid are obtained.

- CN 101537029 reports a method of preparing the extract of the horn powder 33 which is subjected to the yeast hydrolysis; the fermentation lasts 4-8 days, the extract obtained is subjected to ultrasound, filtration and the horn powder thus prepared is subjected to extraction with 75-95% ethanol 35 by ultrasonication, filtration, concentration, obtaining a solid in the form of scales containing ursolic acid. 37

- RO 120950 B1 presents a process for obtaining a bioactive complex of triterpenic acids of at least 90%, starting from the vegetable raw material through a series of extraction steps 39 with a mixture of ethyl alcohol: structured water, followed by concentration, filtration, purification with activated carbon, concentration, precipitation, washing with distilled water, up to neutral pH 41, resulting in a powder which represents the active substance that can be used in the conditioning of medicines for internal use. 43

From the data presented above, regarding the state of the art in the field of obtaining triterpenic acids used as a medicinal substance, by extracting the different sources of 45 plant raw materials, the patents have the following disadvantages:

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- uses numerous light flammable solvents - WO 2005075614 (A1) - hexane, CN 101538271 - chloroform, US 6740778 - hexane, chloroform, US 18186346 - petroleum ether, RU 2108803 - methyl-butyl ether, RU 2151139 - benzene, petroleum ether ; or expensive SU 18186346 isopropyl alcohol;

- most of the processes - SU 18186346, RU 2108803, RU 2151139, CN 1358733, regarding the obtaining of ursolic and oleanolic acids in purified state, does not mention the purity of the substance and its evaluation by quantitative analytical methods (HPLC, GC and others);

- the plant raw materials are different from those used by us in the patent proposal, namely: olive leaves - GR 10011738, US 670778; Oxycoccus quadiipetelus SU 18186346; different species of fir-needles or bark RU 2108107, RU 2108803, RU 2151139 or plants from the Asian area Ligustrum lucidum, // ex kudingcha, Paulownia, Dita, Loquat,

- the processes that claim the obtaining of triterpenic acids concentrates, based on a mixture of ursolic and oleanolic acid from fruit wastes - EP1161879, US 20022037882, EP 1250852, US 2003049365, has the disadvantage of obtaining products with a low content (30 ... 65 %), which cannot be applied in therapeutics, conditioned as medicines, but only as nutritional supplements;

- fruit wastes or fir needles have a low content of triterpenic acids (<0.5%), which negatively affects the profitability of technologies at the industrial level compared to the technologies of processing the usual plant raw materials (medicinal and aromatic plants);

- according to the procedure described in the patent R0120950 B1, the final stage of precipitation and washing with water determines the obtaining of a substance with an amorphous structure, the amorphous state being thermodynamically unstable influences the stability of the structure of the drug substance;

- none of the presented processes has technological data similar to the process which is the subject of the present invention;

- there are no patents and data in the literature related to the correlation between the process of obtaining, the physical-chemical properties, and the bioavailability of triterpenic acids ursolic acid / oleanolic acid, as a medicinal substance.

The analysis of the state of the art in the field of pharmacology of triterpenic acids shows that most of their pharmacological effects have been demonstrated by experimental scientific research whose results have been published in the form of articles or have been the subject of patent applications and patents.

Thus, J. Liu presents the following pharmacological effects evidenced in oleanolic and ursolic acids: anti-inflammatory, hepatoprotective, antihyperlipidemic, antiulcerous, hypoglycemic, antitumor, anti HIV, antimicrobial.

Somova L. I. et al. highlights the cardiovascular, antihypertensive, antihyperlipidemic, antiaterosclerotic and antioxidant activity of oleanolic and ursolic acids isolated from European O / ea.

And Kuttan G. et al. mentions the immunomodulatory activity of ursolic, oleanolic and glycyrrhizic acids.

US patent 4752606 discloses a pharmaceutical composition based on oleanolic acid, as such or as physiologically acceptable salts, conditioned as tablets and usable for prophylactic or curative treatment of various ulcerogenic dysfunctions of the stomach or intestines.

Patent application US 10/488682, published with no. US 2004/0235785 A1, presents a pharmaceutical composition that can be used as a dietary supplement in the treatment or prevention of cancer, as a potentiator of the antitumor agents used in immunotherapy and oncology. According to the invention, the composition is based on terpene compounds (mono-, di-, tri-, sequiterpene).

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Patent application JP 05-211065, published under no. 07-048260 / 1995, 1 shows a bioproduct that can be used as a blood cell growth agent (erythrocytes) based on ursolic acid, given in daily doses of 1-1000 mg and having a 3-life prolongation effect in radiation therapy. different forms of cancer.

Patent applications EP1495754A2, published on 12.01.2005, and EP 0943620 A2, 5 published on 22.09.1999, present a method and composition based on betulinic acid derivatives applicable for preventing or inhibiting the growth of malignancies. There is presented a 7 method of obtaining betulinic acid from the bark of the plant Ziziphus mauritiana (Rhamnacaee fam.), Invitro studies on the cytotoxicity of betulinic acid to different cancer cell lines 9 (melanoma, fibrosarcoma, breast and colon cancer, epidermoid carcinoma) and the results of "in vivo" experiments in mice, which confirm the quality of betulinic acid as an anti-neoplastic agent.

US patent 5985924 discloses a novel metastasis suppressant, 13 with a low degree of toxicity, based on ursolic acid, as such or in the form of salts, which may be administered to patients by bone or injectable, in the postoperative treatment of 15 different forms of cancer.

Patent application JP 63166195, published with no. 02017121 A / 1990, 17 discloses an medicinal product for external use, which can be used to prevent cancer of epithelial cells of the skin, based on ursolic or oleanolic acid, alone or in combination. 19

Patent application JP 09-199323, published with no. JP 11-029467 / 1999, presents a product with inhibitory effects on the protease, based on ursolic acid or its salts 21, applicable for the treatment or prevention of various skin dysfunctions (dermatitis) and conditioned in the form of ointment, cream, lotion and emulsion. The 23 dermatoprotective effects of the product are mentioned: antioxidants, moisturizers and bleaching of the skin.

U.S. Patent No. 4857554 discloses a method for the treatment of psoriasis, 25 based on the daily use of an ointment containing ursolic and oleanolic acids in a 3: 1 ratio, dispersed in a fat base with lanolin and vaseline. The action of the product is demonstrated 27 by clinical experiment for 3 weeks, which revealed that the serious forms of psoriasis were cured in a proportion of 60 ... 70%. 29

The technical problem that the present invention solves consists of:

- elaboration of process conditions through which a bioactive complex with 31 specific design for a certain type of crystalline structure is obtained, which favors its solubility increase at 1: 118 (m: m) or 1: 150 (m: v), resulting in an improvement of the bioavailability 33 of the active substance, which allows to increase the efficacy of the product and a potential widening of the therapeutic area of the drug; 35

- the bioactive complex of triterpenic acids consists in that it contains 100% triterpenic acids, consisting of at least 80% ursolic acid and maximum 20% oleanolic acids, 37 hydroxyursolic, hydroxyoleanolic, betulinic, dehydrobetulinic, in free state or as Na salts. , K or NH ₄ , with a solubility of 1: 118 (m: m) in ethyl alcohol, obtained by extraction from 39 different Salvia species;

- the process of obtaining the bioactive complex of triterpenic acids consists in that 41 that the dried and pulverized vegetable raw material, consisting of Salvia species herba, is extracted by dynamic maceration in the ratio 1/15 ... 1/30 m / v, vegetal mass / solvent, with one of the 43 extraction solvents represented by the mixtures consisting of: 5 ... 15% structured water, "!" with 85 ... 95% acetone or 85 ... 95% ethyl alcohol 95 °, time from 8 ... 24 h, at 15 ... 80 ° C, 45 resulting in the extraction solution, which for purification, is passed on a chromatographic column with activated carbon granulated with acid pH, then concentrated under vacuum at 47

35 ... 40 ° C to 1/10 ... 1/20 of the initial volume, obtaining a suspension that is maintained

RO 127000 Β1 for 12 ... 24 h at 5 ... 10 ° C, then filtered under vacuum, the substance obtained after drying at 105 ° C, for 3 h, constituting the bioactive complex of crude triterpenic acids, with a content of at least 70%, which for purification, dissolves at reflux - 80 ° C, in the ratio 1/100 ... 1/200 m / v, in one of the solvents represented by the mixtures consisting of: 5 ... 10 % structured water "l" or "S" with 90 ... 95% ethyl alcohol 95 ° C or 90 ... 95% acetone; the solution obtained is purified by adsorption on activated carbon with acid pH, added in the ratio of 0.1 ... 0.3% m / v, filtered, concentrated in vacuo at 35 ... 40 ° C to 1 / 10 ... 1/20 from the initial volume, and the resulting suspension is left 12 ... 24 h at 5 ... 10 ° C, filtered under vacuum, the precipitate obtained was washed with 0.3 ... 0.5 The distilled water, to neutral pH, is dried at 105 ° C for a minimum of 3 hours, sprayed to obtain the sprayed product with a concentration of at least 90% triterpenic acid, which is subjected to crystallization from ethyl alcohol, acetone or acetate. ethyl by dissolution at reflux under vacuum, with raising the vapor temperature to 5O ... 6O ° C, concentration under vacuum up to 1/10 ... 1/20 of the initial volume, the crystalline suspension obtained cools rapidly on the ice bath , under stirring, it is filtered immediately under vacuum and dried at 90 ... 110 ° C in hot air, obtaining the bioactive complex of 100% triterpenic acids, of which u rsolic minimum 80% and maximum 20% mixture of oleanolic, hydroxyursolic, hydroxyoleanolic, betulinic, dehydrobetulinic acid, resulting in the active substance for medicinal products for human and / or veterinary use, conditioned in the form of: solution for injection or emulsion for infusion, tablets or capsules soft and / or gelatinous soft, gel, cream or ointment, solution for external use or spray, suppositories or ova.

The advantages of the invention compared to the prior art are the following:

- The process according to the invention is applied at industrial level, using solvents with a medium flammability and vegetable raw materials existing in the national geographical area Salvia officinalis with an average content of triterpenic acids of 3 ... 4%;

- the application of the process of obtaining according to the invention envisages an increase with

10 ... 15% of the extraction yield of triterpenic acids;

- due to the mixture of extraction solvents, a high degree of process selectivity is achieved;

- the purification of the bioactive complex of triterpenic acids ensures an advanced purity of the finished product (100% triterpenic acids), as well as a standardization of the content or in components;

- the modeling stage of the physical structure of the obtained powder ensures the obtaining of the bioactive complex in crystalline form, with the improvement of the properties related to stability, but also to the dissolution process, with maintaining a balance between bioavailability and chemical stability;

- the technological process also exploits other triterpenic acids present in the extractive solutions, which confers an increase of the bioavailability of the whole complex;

- the bioactive complex of triterpenic acids has a 100% standardized content, having a marked and diversified pharmacological activity, exerted at the level of vital functions, namely: activity of bioregulation of the protein metabolism, carbohydrate and lipid, especially on the aged organisms, activity immunoprotective and hepatoprotective, to counteract the immunosuppressive and hepatotoxic adverse effects, after treatment with glucocorticosteroids, cytostatic, antiviral and radiotherapy; tumor retardation activity; local anti-inflammatory activity;

- based on these pharmacological effects, the bioactive triterpenic acid complex, obtained according to the invention, has been conditioned in the form of new medicinal products for human and / or veterinary use such as: capsules, tablets, injectable solutions, oral and external use: spray, gel, cream, ointment, suppositories, ova, intended for new therapeutic applications compared to those currently known.

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The process according to the invention consists in that the dried and 1 sprayed vegetable raw material, with 3 ... 4% triterpenic acids, consisting of Salvia species herba, is extracted by dynamic maceration in the ratio 1/15 ... 1/30 m / v vegetal mass / solvent, with one of the 3 extraction solvents represented by the mixtures consisting of: 5 ... 15% structured water "I" with

85 .. .95% acetone or 85 ... 95% ethyl alcohol 95 °, for 8 ... 24 h, at 15 ... 80 ° C, resulting in 5 extractive solution, which for purification is passed on a chromatographic column with activated carbon granulated with acid pH, then concentrated in vacuo, at 35 ... 40 ° C, up to 7 to 1/10 ... 1/20 of the initial volume, giving a suspension which it is kept for 12 ... 24 h at 5 ... 10 ° C, then filtered under vacuum, the substance obtained after drying at 105 ° C, for 3 h, 9 constituting the bioactive complex of crude triterpenic acids, with a content of at least 70%. for purification, the crude product is dissolved at reflux - 80 ° C - in the ratio 1/100 ... 1/200 m / v in 11 one of the solvents represented by the mixtures consisting of: 5 ... 10% structured water "I "With 90 ... 95% ethyl alcohol 95 ° or 90 ... 95% acetone. 13

The solution obtained is purified by adsorption on activated carbon with acid pH, added in the ratio 0.1 ... 0.3% m / v, filtered, concentrated in vacuo at 35 ... 40 ° C to 1/10. ..1 / 20 15 from the initial volume, the resulting suspension is left 12 ... 24 h at 5 ... 10 ° C, filtered under vacuum, the precipitate obtained is washed with 0.3 ... 0.51 distilled water , to neutral pH, is dried at 105 ° C for a minimum of 3 hours and sprayed to obtain the sprayed product with a concentration of at least 80% triterpenic acid, which is subjected to crystallization from ethyl alcohol, acetone or 19 ethyl acetate. by dissolving under reflux under vacuum, with raising the vapor temperature to 50 ... 60 ° C, concentrating under vacuum up to 1/10 -1/20 of the initial volume, the crystalline suspension obtained is cooled to an ice bath rapidly, below shake, filter immediately under vacuum and dry at

90 .. .110 ° C in hot air flow, obtaining the bioactive complex of 100% triterpenic acids, 23 of which ursolic acid minimum 80% and maximum 20% mixture of oleanolic, hydroxyursolic, hydroxyoleanolic, betulinic, dehydrobetulinic acid. 25

To obtain the complex of triterpenic acids in the form of Na, K, or salts

NH ₄ , the product sprayed with a concentration of at least 80% triterpenic acids dissolves in the ratio 27 1/100 ... 1/200 m / v in the solvent mixture with alkaline pH: 90 ... 95% ethyl alcohol 95 ° with 5 ... 10% structured water "S", which also contains 1% NaOH or 1% KOH or 5% NH ₄ OH 25%, 29 followed by the concentration of the resulting solution and obtaining the bioactive complex of triterpenic acids as Na salt, K or NH ₄ , which is subjected to crystallization from ethyl alcohol, acetone 31 or ethyl acetate by dissolution at reflux under vacuum, raising the vapor temperature to

50 .. .60 ° C, concentration in vacuo to 1/10 ... 1/20 of the initial volume, the crystalline suspension 33 obtained is cooled on an ice-bath rapidly, with stirring, filtered immediately under vacuum and dried at 90 ... 110 ° C in hot air stream, obtaining the bioactive complex of 35 triterpenic acids as Na, K or NH ₄ salt, with a content of at least 90%, evaluated and a phase yield of 95 %. 37 further described are examples of embodiments of the invention, which refer to the process for obtaining the bioactive complex of triterpenic acids and to the conditioning drug products 39.

Example 1.41

Variant A. Obtaining the bioactive complex of triterpenic acids on an industrial scale from grass Salvia species (Salvia) 43 kg dry and pulverized vegetable raw material, consisting of grass Salvia species (Salvia officinalis, Salvia lavandulifolia, Salvia triloba), with a content of 3. .4% triterpenic acids45, are extracted with 500 l of extraction solvent consisting of: 90 volumes (450 I) acetone + 10 volumes (501) structured water ,,! ”, CupH = 1.8 ... 2.6, by dynamic maceration, for 12 hours, 47 at ambient temperature (15 ... 30 ° C), after which a minimum 400 I solution is collected

RO 127000 Β1 extract which is passed on a chromatographic column with 2 kg of acidic granulated activated carbon, resulting in a purified extractive solution, which is concentrated under vacuum at room temperature.

35 ... 40 ° C, up to the volume of 40 I, obtaining a suspension which is kept for 24 hours at 5 ... 10 ° C, then filtered under vacuum and the precipitate on the filter is dried at 105 ° C, for 3 hours and sprayed. At least 1200 g of light greenish yellow powder is obtained, which is the bioactive complex of crude triterpenic acids, with a content of at least 70%.

For the purpose of product purification, the solvent mixture with acid pH consisting of 90 volumes (1081) ethyl alcohol 95 ° + 10 volumes (12 I) structured water, "!", with pH = 1.8 ... 2.6, is used. to which is added, for dissolution, 1200 g of powder consisting of the bioactive complex of crude triterpenic acids (calculated for 1 g of substance -100 ml of solvent mixture); the suspension is heated to 80 ° C for 1 h until the substance is completely dissolved, and the resulting yellow solution is treated with 0.1 ... 0.3% m / v acidic charcoal; The slurry is filtered, the solution obtained is concentrated in vacuo at 35.40 ° C to 1/10 of the initial volume (121), when a microcrystalline slurry results, which is maintained for 24 hours at 5. .. 10 ° C, then filtered under vacuum, the precipitate was washed with 0.5 l of distilled water, until neutral pH, dried at 105 ° C, for 3 h and sprayed, which constitutes the complex. triterpenic acids, with a content of at least 80%, it is subjected to crystallization from organic solvents, ethyl acetate by dissolution at reflux under vacuum, with raising the vapor temperature to 5O ... 6O ° C, concentration under vacuum up to 1 / 10 -1/20 of the initial volume, the crystalline suspension obtained is cooled on an ice bath rapidly, with stirring, immediately filtered under vacuum and dried at 90 ... 110 ° C in hot air stream, yielding 600 g white crystalline substance, which represents the bioactive complex of triterpenic acids 10 0%, of which ursolic acid minimum 80% and maximum 20% mixture of other triterpenic acids, consisting of one or more of the acids: hydroxyursolic oleanolic acid, hydroxyoleanolic acid, betulinic, dehydrobetulinic, which represents a final yield of at least 50% reported to the triterpenic acid content of the processed vegetable raw material.

Variant B.

kg raw plant material, dried and sprayed, consisting of grass Salvia species (Salvia officinalis, Salvialavandulifolia, Salvia triloba, Salvia sclarea etc.), with a content of 3-4% triterpenic acids, extracted with 5001 extraction solvent, consisting of : 90 volumes (450 I) ethyl alcohol 95 ° + 10 volumes (50 I) structured water "I" (pH = 1.8 ... 2.6), by dynamic maceration, for 12 hours, at ambient temperature ( 15 ... 30 ° C), after which a minimum of 4001 extraction solution is collected, which is passed on a chromatographic column with 2 kg of acidic granulated activated carbon, resulting in a purified extractive solution, which is concentrated in vacuo, at 35. ..40 ° C, up to the volume of 40 I, obtaining a suspension which is kept 24 h at 5 ... 10 ° C, after which it is filtered under vacuum and the precipitate on the filter is dried at 105 ° C for 3 hours and sprayed. At least 1200 g of yellow powder is obtained, slightly greenish, which is the complex of crude triterpenic acids, with a content of at least 70%.

For the purification of the crude product, the solvent mixture with acid pH, consisting of 90 volumes (108 I) acetone +10 volumes (12 I) structured water "I" with a pH = 1.8 ... 2.6, is used in which is added, for dissolution, 1200 g powder consisting of the crude triterpenic acid complex (calculated for 1 g substance -100 ml solvent mixture); the slurry is heated at 80 ° C for 1 h, until the substance is completely dissolved, and the resulting yellow solution is treated with 0.1 ... 0.3% m / v acidic charcoal; the suspension is filtered, the solution obtained is concentrated in vacuo, at a temperature of 35 ... 40 ° C, up to 1/10 of the initial volume (12 I), when a microcrystalline suspension is obtained, which is kept for 24 hours at

RO 127000 Β1 temperature of 5 ... 10 ° C to complete the crystallization, after which it is filtered under vacuum, 1 the precipitate is washed with 0.5 I distilled water, until neutral pH, dried at room temperature.

105 ° C, for 3 hours, is sprayed, it constitutes the complex of purified triterpenic acids 3 with a content of at least 80%, which is subjected to crystallization from organic solvents, ethyl acetate by dissolution under reflux under vacuum, with raising the temperature. of vapors at 50 ... 60 ° C, 5 concentration under vacuum up to 1/10 ... 1/20 of the initial volume, the crystalline suspension obtained is cooled on an ice bath rapidly, with stirring, filtered immediately under vacuum and dry at 7

90 ... 110 ° C in hot air stream, yielding 610 g of white crystalline substance, which represents the bioactive complex of 100% triterpenic acids, of which ursolic acid minimum 81% and maximum 9 19% mixture of other triterpenic acids, consisting of one or more of the acids: oleanolic hydroxyursolic acid, hydroxyoleanolic, betulinic, dehydrobetulinic acid, which represents a final final yield of at least 50% based on the triterpenic acid content of the processed plant raw material. 13

Example 2. Obtaining the bioactive complex of triterpenic acids as Na 15 salt

200 g powder of bioactive complex purified of triterpenic acids, with a content of at least 80%, obtained according to the working method described in embodiment 1, is dissolved in 20 l (ratio 1/100 m / v) solvent mixture consisting of : 90 volumes (18 L) ethyl alcohol 95 ° + 10 volumes (2 L) structured 'S' water (pH = 10 ... 12), containing 1% NaOH, through 19 reflux heating (80 ° C), obtaining a clear solution which is concentrated in vacuo at 4O ... 45 ° C, up to 1/4 of the initial volume (5 I), resulting in a 21 microcrystalline suspension, which is maintained 24 hours, at 5 ... 10 ° C, in order to complete the crystallization, after which it is filtered under vacuum, the precipitate on the filter is washed with minimum 0.1 I 23 distilled water to neutral pH, dried at 105 ° C for 3 h and is sprayed and subjected to crystallization from organic solvents, ethyl acetate by dissolving 25 under reflux under vacuum, with raising temples vapor ratios at 5O ... 6O ° C, vacuum concentration up to 1/10 ... 1/20 of the initial volume, the crystalline suspension obtained is cooled on ice bath 27 rapidly, with stirring, filtered immediately under vacuum and is dried at 90 ... 110 ° C in hot air, yielding 180 g of white crystalline substance, which represents the bioactive complex of 29 triterpenic acids in the form of Na salt of ursolic, oleanolic and other triterpenic acids, consisting of one or more of the acids: hydroxyursolic, hydroxyoleanolic, betulinic, 31 dehydrobetulinic, with a content of at least 90%, which represents a phase yield of 95%. 33

Example 3. Obtaining the bioactive complex of triterpenic acids in the form of K 35 salt

200 g purified bioactive complex of triterpenic acids, with a content of at least 80%, 37 obtained according to the working method described in embodiment 1, is dissolved in 20 I solvent mixture consisting of: 90 volumes (18 I) ethyl alcohol 95 ° + 10 volumes (2 L) structured water "S" (pH = 10 ... 12), containing 1% KOH, by reflux heating (80 ° C); The resulting clear solution is concentrated in vacuo at 4O ... 45 ° C, up to 1/4 of the initial volume (5 I), resulting in a microcrystalline suspension, which is maintained 24 hours, at 5 ... 10 ° C, in order to complete the crystallization, after which it is filtered in vacuo, the precipitate on the filter is washed with a minimum of 0.1 I distilled water to neutral pH, dried at 105 ° C for 3 h, pulverizes and is subjected to crystallization from 45 organic solvents, ethyl acetate by dissolution at reflux under vacuum, with raising the vapor temperature to 50 ... 60 ° C, concentration under vacuum up to 1/10 ... 1/20 of the initial volume, the crystalline suspension 47 obtained is cooled on an ice-bath rapidly, with stirring, filtered immediately under vacuum and dried at 90 ... 110 ° C in hot air, yielding 180 g of white substance.

RO 127000 Β1 crystalline representing the bioactive complex of triterpenic acids in the form of K salt of ursolic, oleanolic acids and other triterpenic acids, consisting of one or more of the acids: hydroxyursolic, hydroxyoleanolic, betulinic, dehydrobetulinic, with a minimum content

90%, which represents a phase yield of 95%.

Example 4. Obtaining the bioactive complex of triterpenic acids in the form of NH ₄ salt

200 g bioactive complex of purified triterpenic acids, with a content of at least 80%, obtained according to the working method described in embodiment 1, is dissolved in 20 I solvent mixture consisting of: 90 volumes (18 I) ethyl alcohol 95 ° + 10 volumes (2 I) "S" structured water (pH = 10 ... 12), containing 5% NH ₄ OH 25%, by reflux heating (80 ° C); The resulting clear solution is concentrated in vacuo at 4O ... 45 ° C, up to 1/4 of the initial volume (5 I), resulting in a microcrystalline suspension, which is kept 24 hours, at 5 ... 10 ° C, in order to complete the crystallization, after which it is filtered under vacuum, the precipitate on the filter is washed with a minimum of 0.1 L distilled water to neutral pH, dried at 105 ° C for 3 h, sprayed and is subjected to crystallization from organic solvents, ethyl acetate by dissolution at reflux under vacuum, with raising the vapor temperature to 50 ... 60 ° C, concentration under vacuum up to 1/10 ... 1/20 of the initial volume, the crystalline suspension obtained is cooled on an ice-bath rapidly, with stirring, filtered immediately under vacuum and dried at 90 ... 110 ° C in hot air, yielding 180 g of white crystalline substance, which represents the bioactive complex of triterpenic acids in the form of NH ₄ salt of ursolic acids, olean olic and other triterpenic acids, consisting of one or more of the acids: hydroxyursolic, hydroxyoleanolic, betulinic, dehydrobetulinic, with a content of at least 90%, which represents a phase yield of 95%.

Example 5. Variant A. Obtaining the bioactive complex of triterpenic acids on the industrial scale of Sal via species (Salvia) kg dry and pulverized vegetable raw material, consisting of Salvia species (Salvia officinalis, Salvia lavandulifolia, Salvia triloba), with a content of 3 ... 4% triterpenic acids, are extracted with 500 I extraction solvent consisting of: 90 volumes (450 I) acetone + 10 volumes (501) structured water "I", with pH = 1.8 ... 2, 6, by dynamic maceration, for 12 hours, at ambient temperature (15 ... 30 ° C), after which at least 400 l of extractive solution is collected which is passed on a chromatographic column with 2 kg of acid-granulated activated carbon, resulting in a purified extractive solution, which is concentrated in vacuo at room temperature

35 ... 40 ° C, up to the volume of 40 I, obtaining a suspension which is kept for 24 hours at 5 ... 10 ° C, then filtered under vacuum and the precipitate on the filter is dried at 105 ° C, for 3 hours and sprayed. At least 1200 g of light greenish yellow powder is obtained, which is the bioactive complex of crude triterpenic acids, with a content of at least 70%.

For the purpose of product purification, the solvent mixture with acid pH consisting of 90 volumes (108 I) ethyl alcohol 95 ° + 10 volumes (12 I) structured water "I", with pH = 1.8 ... 2.6 is used. , to which is added, for dissolution, 1200 g of powder consisting of the bioactive complex of crude triterpenic acids (calculated for 1 g of substance -100 ml of solvent mixture); the suspension is heated to 80 ° C for 1 h until the substance is completely dissolved, and the resulting yellow solution is treated with 0.1 ... 0.3% m / v acidic charcoal; the suspension is filtered, the solution obtained is concentrated in vacuo, at a temperature of 35, 40 ° C, up to 1/10 of the initial volume (12 I), when a microcrystalline suspension is obtained, which is maintained for 24 hours at 5 ... 10 ° C, then filtered under vacuum, the precipitate was washed with 0.5 l of distilled water, until neutral pH, dried at 105 ° C for 3 hours and sprayed, which constitutes the complex of triterpenic acids, with a minimum content

RO 127000 Β1

80%, it is subjected to a crystallization from organic solvents, ethyl alcohol by dissolution at 1 reflux under vacuum, with raising the vapor temperature to 5O ... 6O ° C, concentration under vacuum up to 1/10 ... 1/20 from the initial volume, the crystalline suspension obtained is cooled on an ice bath 3 rapidly, with stirring, immediately filtered under vacuum and dried at 9O ... 11O ° In hot air stream, obtaining 550 g of white crystalline substance, which represents the bioactive complex of 5 triterpenic acids 100%, of which ursolic acid minimum 80% and maximum 20% mixture of other triterpenic acids, consisting of one or more of the acids: hydroxyursolic oleanolic acid, 7 hydroxyoleanolic acid, betulinic, dehydrobetulinic acid, which represents a final yield of at least 50% based on the triterpenic acid content of the processed plant raw material. 9 Variant B. 30 kg plant raw material, dried and sprayed, consisting of grass

Salvia species (Salvia officinalis, Salvia lavandulifolia, Salvia triloba, Salvia sclarea etc), with 11 containing 3 ... 4% triterpenic acids, are extracted with 5001 extraction solvent, consisting of:

volumes (450 I) ethyl alcohol 95 ° + 10 volumes (50 I) structured water "I" (pH = 1.8 ... 2.6), through 13 dynamic maceration, for 12 hours, at ambient temperature (15 ... 30 ° C), after which a minimum of 400 l of extractive solution is collected, which is passed on a chromatographic column with 15 kg of acidic granulated activated carbon, resulting in a purified extractive solution, which is concentrated in vacuo, at a temperature of 35. ..40 ° C, up to the volume of 40 I, obtaining a super-17 pension which is maintained 24 hours at 5 ... 10 ° C, after which it is filtered under vacuum, and the precipitate on the filter is dry at 105 ° C for 3 hours and spray. A minimum of 19 1200 g of yellow powder is obtained, slightly greenish, which constitutes the complex of crude triterpenic acids, with a content of at least 70%. 21

For the purification of the crude product, the mixture of solvents with acid pH, consisting of 90 volumes (108 I) acetone + 10 volumes (12 I) structured water "I" with a pH 23 = 1.8 ... 2.6 in which is added, for dissolution, 1200 g powder consisting of the crude triterpenic acid complex (calculated for 1 g substance -100 ml solvent mixture); The suspension is heated to 80 ° C for 1 h, until the substance is completely dissolved, and the resulting yellow solution is treated with 0.1 ... 0.3% m / v acidic charcoal; the suspension is filtered, the solution obtained is concentrated in vacuo, at a temperature of 35 ... 40 ° C, up to 1/10 of the initial volume (12 I), when a microcrystalline suspension is obtained, which is maintained for 24 hours at 29 from 5 to 10 ° C, to complete the crystallization, then filtered under vacuum, the precipitate was washed with 0.5 l of distilled water, until neutral pH, dried at 31 105 ° C, for 3 h. , it is sprayed, it constitutes the complex of purified triterpenic acids, with a content of at least 80%, which is subjected to crystallization from organic solvents, ethyl alcohol 33 by dissolution at reflux under vacuum, raising the vapor temperature to 50 ... 60 ° C, concentration in vacuo to 1/10 ... 1/20 of the initial volume, the crystalline suspension obtained is cooled rapidly on ice bath, with stirring, immediately filtered under vacuum and dried at

90 ... 110 ° C in hot air stream, yielding 560 g of white crystalline substance, representing 37 bioactive complex of 100% triterpenic acids, of which ursolic acid minimum 81% and maximum 19% mixture of other triterpenic acids, consisting of one or more of the acids: 39-oleanolic hydroxyursolic acid, hydroxyoleanolic, betulinic, dehydrobetulinic acid, which represents a final yield of at least 50% based on the triterpenic acid content of the processed vegetable raw material 41.

Example 6. Variant A. Obtaining the bioactive complex of triterpenic acids on industrial scale 43 from grass Salvia species (Salvia) kg dry and pulverized vegetable raw material, consisting of grass Salvia 45 species (Salvia officinalis, Salvia lavandulifolia, Salvia triloba etc.), with with a content of 3 ... 4% triterpenic acids, is extracted with 500 I extraction solvent consisting of: 90 volumes (450 I) 47 acetone + 10 volumes (501) structured water "I", cupH = 1.8. .2.6, by dynamic maceration, time

RO 127000 Β1 of 12 h, at ambient temperature (15 ... 30 ° C), after which a minimum of 4001 extraction solution is collected, which is passed on a chromatographic column with 2 kg of acid granulated activated carbon, resulting in a purified extractive solution, which is concentrated in vacuo at

35 .. .40 ° C, up to the volume of 40 I, obtaining a suspension which is maintained for 24 hours at 5 ... 10 ° C, then filtered under vacuum and the precipitate on the filter is dried at 105 ° C, for 3 hours and sprayed. At least 1200 g of light greenish yellow powder is obtained, which is the bioactive complex of crude triterpenic acids, with a content of at least 70%.

For the purpose of product purification, the solvent mixture with acid pH, consisting of 90 volumes (1081) ethyl alcohol 95 ° + 10 volumes (12 I) structured water "I", with pH = 1.8 ... 2.6 is used. , to which is added, for dissolution, 1200 g of powder consisting of the bioactive complex of crude triterpenic acids (calculated for 1 g of substance -100 ml of solvent mixture); the suspension is heated to 80 ° C for 1 h until the substance is completely dissolved, and the resulting yellow solution is treated with 0.1 ... 0.3% m / v acidic charcoal; the suspension is filtered, the solution obtained is concentrated in vacuo, at a temperature of 35, 40 ° C, up to 1/10 of the initial volume (12 I), when a microcrystalline suspension is obtained, which is maintained for 24 hours at 5 ... 10 ° C, then filtered under vacuum, the precipitate was washed with 0.5 l of distilled water, until neutral pH, dried at 105 ° C for 3 hours and sprayed, which constitutes the complex of triterpenic acids, with a content of at least 80%, it is subjected to a crystallization of acetone by dissolution at reflux under vacuum, raising the vapor temperature to

50 .. .60 ° C, concentration in vacuo to 1/10 ... 1/20 of the initial volume, the crystalline suspension obtained is cooled on an ice-bath rapidly, with stirring, filtered immediately under vacuum and dried at 90 ... 110 ° C in hot air stream, yielding 580 g white crystalline substance, which represents the bioactive complex of 100% triterpenic acids, of which ursolic acid minimum 80% and maximum 20% mixture of other triterpenic acids, consisting of one or more of the acids: hydroxyursolic, hydroxyoleanolic, betulinic, dehydrobetulinic oleanolic acid, which represents a final yield of at least 50% based on the triterpenic acid content of the processed plant raw material.

Variant B. 30 kg plant raw material, dried and sprayed, consisting of grass Salvia species (Salvia officinalis, Salvia lavandulifolia, Salvia triloba, Salvia sclarea etc.), with a content of 3 ... 4% triterpenic acids, extracted with 5001 extraction solvent, consisting of: 90 volumes (450 I) ethyl alcohol 95 ° + 10 volumes (50 I) structured water "I" (pH = 1.8 ... 2.6), by dynamic maceration, during 12 h, at ambient temperature (15 ... 30 ° C), after which a minimum of 400 l of extractive solution is collected, which is passed on a chromatographic column with 2 kg of acid granulated activated carbon, resulting in a purified extractive solution, which is concentrated under vacuum, at a temperature of 35 ... 40 ° C, up to a volume of 401, giving a suspension which is kept for 24 hours at 5 ... 10 ° C, after which it is filtered under vacuum and the precipitate the filter is dried at 105 ° C for 3 hours and sprayed. At least 1200 g of yellow powder is obtained, slightly greenish, which is the complex of crude triterpenic acids, with a content of at least 70%.

For the purification of the crude product, the solvent mixture with acid pH, consisting of 90 volumes (108 I) acetone +10 volumes (12 I) structured water "I" with a pH = 1.8 ... 2.6, is used in which is added, for dissolution, 1200 g powder consisting of the crude triterpenic acid complex (calculated for 1 g substance -100 ml solvent mixture); the slurry is heated at 80 ° C for 1 h, until the substance is completely dissolved, and the resulting yellow solution is treated with 0.1 ... 0.3% m / v acidic charcoal; the suspension is filtered, the solution obtained is concentrated in vacuo, at a temperature of 35 ... 40 ° C, up to 1/10 of the initial volume (12 I), when a microcrystalline suspension is obtained, which is kept for 24 hours at

RO 127000 Β1 temperature of 5 ... 10 ° C to complete the crystallization, after which it is filtered under vacuum, the precipitate is washed with 0.5 I distilled water, to neutral pH, dried at 105 ° C , for 3 hours, it is sprayed, it constitutes the complex of purified triterpenic acids, with a 3 content of at least 80%, which is subjected to a crystallization of acetone by dissolution under reflux under vacuum, raising the vapor temperature to 5O ... 6O ° C, concentration under vacuum up to 5 1/10 ... 1/20 of the initial volume, the crystalline suspension obtained is cooled on an ice-bath rapidly, with stirring, immediately filtered under vacuum and dried at 9O ... 11O ° In hot air stream, 7 yielding 590 g white crystalline substance, representing the bioactive complex of 100% triterpenic acids, of which ursolic acid minimum 81% and maximum 19% mixture of other 9 triterpenic acids, consisting of one or several acids: hydroxyursolic oleanolic acid, hydroxyoleanolic, betulinic, dehydrobetulinic, which represents a final yield of at least 50% based on the triterpenic acid content of the processed plant raw material.

Examples of conditioning of the bioactive complex of triterpenic acids in the form of 13 medicinal products based on the results of the pharmacological tests performed, the bioactive complex of 15 triterpenic acids as such or in the form of Na, K or NH ₄ salts can be conditioned in the form of medicinal products for internal or external use, intended for clinical areas with wide social incidence belonging to human or veterinary therapists. The medicinal products described below, according to the invention, are administered in human or 19 veterinary therapeutics, by injection, oral or local route, for 1 day ... 24 months, depending on the evolution of the disease, in daily doses of 10 ... 1000 mg / kg. mc, divided into 1 ... 6 prizes. The examples of formulations 21 presented below do not exclude the possibility of making other conditioning formulas, given the diversity and the multitude of excipients known and used in the pharmaceutical industry 23.

Example 7. Condensed medicinal product in capsule form containing 25 100 mg bioactive triterpenic acid / capsule complex

Packaging formula / capsule: 27

- bioactive complex of triterpenic acids - 100% ...................... 105mg

- lactose monohydrate .......................................... 150mg 29

- starch sodium glycolate ...................................... 9.0mg

- polyvinylpyrrolidone K30 ........................................ 6.5 mg31

- sodium lauryl sulfate ......................................... 2.0mg

- aerosil .................................................... ... 1.47 mg33

- talc ................................................ ..... 7.25mg

- magnesium stearate ......................................... 2.9 mg35

Method of preparation: The bioactive complex of triterpenic acids is located with the excipients: lactose monohydrate and aerosil on a sieve of size 11. Over the mixture obtained, pour a solution made by dissolving polyvinylpyrrolidone K30 in ethyl alcohol, and homogenize.

The granules are obtained which are dried at a temperature of 35 ... 45 ° C and then reduced to 39 the size conferred by the sieve V; add sodium starch glycolate, sodium lauryl sodium sulfate, talc and magnesium stearate, homogenize and the powder thus obtained is encapsulated in capsules of size 1. The product is administered in doses of 3 ... 6 capsules / day, for 1 ... 24 months, having the following pharmacological effects: antitumor, immunoprotective, hepa- 43 toprotector, antiulcer, hypoglycemic, hypolipidemic, antiaterosclerotic, anti-inflammatory, vascular toning, especially at coronary level, with therapeutic uses in the following 45 diseases: neoplasm, immune system depletion, acute and chronic hepatitis, liver cirrhosis, ulcer disease, diabetes, cardiovascular disorders with atherosclerotic etiology, degeneration of 47 vascular collagen, dermal tissues: psoriasis, neurodermitis, radiodermitis, varicose syndrome

RO 127000 ale1 and the various manifestations of chronic venous insufficiency, retinal vascular diseases, applicable also in geriatrics to prevent and slow down aging processes.

Example 8. Medicated product in the form of tablets containing

100 mg bioactive complex of triterpenic acids / tablet

Conditioner / tablet formula:

- bioactive complex of triterpenic acids-100% .................... 105.00mg

- lactose monohydrate ........................................ 81.20mg

- microcrystalline cellulose ..................................... 74.00mg

- corn starch ........................................ 16.20mg

- starch sodium glycolate ...................................... 5.00mg

- polividone ................................................ .5,60mg

- magnesium stearate ........................................ 2.00mg

- aerosil .................................................... ... 1,00mg

- talc ................................................ ...... 5,00mg

Method of preparation: the usual procedures known for conditioning the pharmaceutical products for internal use in the form of tablets on industrial level installations are applied, in compliance with the rules of good manufacturing practice for medicines, provided by the European legislation.

The product is administered in doses of 3 ... 6 tablets / day, for 1 ... 24 months, having the following pharmacological effects: antitumor, immunoprotective, hepatoprotective, antiulcerous, hypoglycemic, hypolipidemic, antiaterosclerotic, anti-inflammatory, vascular toning, especially at coronary level, with therapeutic uses in the following diseases: neoplasm, immune system depletions, acute and chronic hepatitis, liver cirrhosis, ulcer disease, diabetes, cardiovascular disorders with atherosclerotic etiology, degeneration of vascular, articular and dermal collagen: psoriasis, psoriasis, radiodermitis, varicose syndrome and various manifestations of chronic venous insufficiency, retinal vascular diseases, applicable also in geriatrics to prevent and slow down aging processes.

Example 9. Gel-conditioned drug product - dermatological use containing 1% bioactive complex of triterpenic acids - Na salt

Conditioning formula:

- bioactive complex of triterpenic acids - minimum 90% Na salt ....... 10.5g

- Carbopol 940 ............................................... .. 20g

- Glycerin ................................................ 60g .....

- Sodium hydroxide solution 15% .................................... 10g

- propylene glycol ................................................ .60g

- Methyl p-hydroxybenzoate ......................................... 1g

- propyl p-hydroxybenzoate ........................................ 2g

- structured water "I" (pH = 1.8 ... 2.7) ............................ ad. 1000 g

Method of preparation: The gel mass is prepared by dispersing 20 g Carbopol 940 in the mixture consisting of 60 g glycerin and 837 g structured water "I", in which the preservative mixture mentioned in the formula is dissolved previously. Shake gently and leave to stand at 15 ... 25 ° C, then add the mixture consisting of 10.5 g of triterpenic acid bioactive complex, 60 g of propylene glycol and 10 g of 15% sodium hydroxide solution under slow stirring. adjusting the pH to around 7.0 and continuing stirring for a good homogenization for 30 minutes, then packing in tubes containing 50 g / tube.

The product contains 1% bioactive complex of triterpenic acids - Na salt and is administered in doses of 10 ... 30 mg / day, for 1 ... 6 months, having the following pharmacological effects: trophic and regenerative of vascular collagen, tissue , joint and dermal, anti-inflammatory,

RO 127000 Β1 antihyaluronidase and antielastase, with therapeutic uses in the following diseases: burns, 1 post-burn and post-operative keloids, ladders, atonic wounds of different etiologies, degeneration of dermal, vascular, tissue and joint collagen such as: benign conditions and malignancies of 3 skin, psoriasis, inflammatory and pruritic dermatoses, neurodermitis, radiodermitis, varicose and post-thrombotic syndrome, chronic venous insufficiency, vascular fragility, 5 rheumatic inflammatory states, as well as diabetic or endocrine obesity.

Example 10. Cream-based drug product containing 1% 7 bioactive triterpenic acid complex - Na salt

Conditioning formula: 9

- bioactive complex of triterpenic acids - minimum 90% Na salt ....... 10.5g

- cetyl alcohol ............................................... 200 g11

- White Vaseline ............................................... 200g

-glycerin ................................................ ... 120 g13

- Tween 80 ............................................... .... 70g

- Methyl p-hydroxybenzoate ....................................... 1g15

- propyl p-hydroxybenzoate ....................................... 2g

- structured water "I" (pH = 1.8 ... 2.7) ........................... ad. 1000g17

Method of preparation: dissolve 10.5 g bioactive complex of triterpenic acids - salt

Na, in 120 g glycerin, and the resulting solution is incorporated into the mass of ointment, obtained by 19 melting at 60 ° C of cetyl alcohol and vaseline, followed by emulsification by adding 70 g of Tween 80 and at least 400 g of structured water "I ”In which 21 preservatives mentioned in the formula were dissolved previously. The mass of ointment is stirred for emulsification and homogenization, then packaged in tubes of 50 g. 23

The product is in the form of creamy, homogeneous, white cream, with rapid absorption, containing 1% bioactive complex of triterpenic acids as Na salt, and is administered 25 in doses of 10 ... 30 mg / day, for 1. ..6 months, having the following pharmacological effects: trophic and regenerative of vascular, tissue, articular and dermal collagen, anti-inflammatory, 27 antihyaluronidase and anti-elastase, with therapeutic uses in the following diseases: burns, post-burn keloids and post-operative, stairs, atonic wounds of different etiologies, degeneration of dermal, vascular, tissue and joint collagen such as: benign and malignant skin disorders, psoriasis, inflammatory and pruritic dermatoses, neurodermitis, radiodermitis, varicose and post-thrombotic syndrome, chronic venous insufficiency vascular fragility, rheumatic inflammatory conditions, as well as diabetic or endocrine obesity.33

Example 11. Medicinal product conditioned in the form of suppositories containing% bioactive complex of triterpenic acids35

Conditioning formula:

- bioactive complex of 100% triterpenic acids ........................ 1.05 g37

- polyethylene glycol 400 ............................................. 5g

- methyl para-hydroxybenzoate .................................... 0.1 g39

- propyl para-hydroxybenzoate ................................... 0.03g

- Suppocire (mixture of semisynthesis triglycerides) ............... ad. 100 g 41

Method of preparation: the bioactive complex of triterpenic acids is dissolved in polyethylene glycol, by heating to 80 ° C. The solution obtained is cooled to 40 ° C and is added over table 43 of Suppocire which has been previously heated to 40 ° C. Add the mixture of preservatives, mix and pour into the suppositories, each weighing 2.5 g.

The product contains 25 mg bioactive complex of triterpenic acids / suppository and can be administered in doses of 50 ... 100 mg / day, for 1 ... 6 months, having the following pharmacological-47 logical effects: immunoprotective, anti-inflammatory, trophic and regenerator of vascular, tissue and collagen

RO 127000 Β1 articular, with uses in the following diseases: immunosuppressive, especially hepatotoxic, occurring after treatment with glucocorticosteroids, cytostatic, inflammatory rheumatic diseases, hemorrhoidal syndrome.

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CN 1453289/2003 - Wang Junxian et al. 3

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- An Encyclopedia of Chemicals, Drugs, and Biologicals ”O'Neil, Maryadele. J. - 2006, p. 169911

Claims (7)

claims 1. Bioactive complex of triterpenic acids, obtained from Salvia officinalis, characterized in that it has a crystalline structure and a solubility of 1: 118 (m: m) or 1: 150 (m: v) in ethyl alcohol, consisting of a minimum of 80 % ursolic acid and maximum 20% mixture consisting of one or more acids: oleanolic, hydroxyursolic, hydroxyoleanolic, betulinic, dehydrobetulinic, obtained as free acids or as salts of Na, K or NH ₄ . 2. Bioactive complex of triterpenic acids, according to claim 1, characterized in that it is obtained and used in the form of salts of Na, K or NH ₄ , of minimum 90% purity, consisting of: minimum 75% ursolate of Na or K or NH ₄ , 10 ... 15% Na or K or oleanolatde NH ₄ and 4 ... 10% Na salts, K or NH ₄ from the other triterpene acids consisting of one or more of the following acids: hydroxyursolic, hydroxyoleanolic, betulinic, dehydrobetulinic. 3. Process for obtaining the bioactive complex of triterpenic acids, defined in claim 1, characterized in that the dried and pulverized vegetable raw material, consisting of Salvia species herba, is extracted by dynamic maceration in the ratio 1/15 ... 1/30 m / v, vegetal mass / solvent, with one of the extraction solvents represented by the mixtures consisting of: 5 ... 15% structured water "I" with 85 .. .95% acetone or 85 ... 95% ethyl alcohol 95 °, for 8 ... 24 h, at 15 ... 80 ° C, resulting in the extraction solution, which for purification, is passed on a chromatographic column with activated carbon granulated with acid pH, then concentrated under vacuum to temperature from 35 ... 40 ° C to 1/10 ... 1/20 of the initial volume, obtaining a suspension which is maintained for 12 ... 24 h at 5 ... 10 ° C, then filter under vacuum, the substance obtained after drying at 105 ° C, for 3 hours, constituting the bioactive complex of crude triterpenic acids with a content of minimum 70%, which for purification, dissolves at reflux - 80 ° C in the ratio 1/100 ... 1/200 m / v in one of the solvents represented by the mixtures consisting of: 5 ... 10% structured water "L" or "S" with 90 ... 95% ethyl alcohol 95 ° or 90 ... 95% acetone; the solution obtained is purified by adsorption on activated carbon with acid pH, added in the ratio of 0, 1 ... 0,3% m / v, filtered, concentrated in vacuo at 35 ... 40 ° C up to 1/10. .1 / 20 of the initial volume and the resulting suspension is left 12 ... 24 h at 5 ... 10 ° C, filtered under vacuum, the precipitate obtained is washed with 0.3 ... 0.51 distilled water, to neutral LapH, dry at 105 ° C for a minimum of 3 hours, spray, obtaining the purified product with a concentration of at least 90% triterpenic acid, which is subjected to crystallization from ethyl alcohol, acetone or ethyl acetate by reflux dissolution. under vacuum, with the vapor temperature rising to 5O ... 6O ° C, vacuum concentration up to 1/10 ... 1/20 of the initial volume, the crystalline suspension obtained is cooled on an ice bath rapidly, with stirring, if filter immediately under vacuum and dry at 90 ... 110 ° C in hot air stream, obtaining the bioactive complex of 100% triterpenic acids, of which ursolic acid c minimum 80% and maximum 20% mixture of oleanolic, hydroxyursolic, hydroxyoleanolic, betulinic, dehydrobetulinic acid. Process according to claim 3, characterized in that the bioactive complex of triterpenic acids crystallized with a content of at least 90% dissolves at reflux at 80 ° C in the ratio 1/100 ... 1/200 m / v in the solvent mixture. with alkaline pH: 90 ... 95% ethyl alcohol 95 ° with 5 ... 10% structured "S" water, with pH = 10 ... 12, which also contains 1% NaOH or 1% KOH or 5% NH ₄ OH 25%, the solution obtained is concentrated in vacuo at 35 ... 40 ° C up to 1/5 ... 1/10 of the initial volume, the resulting suspension is maintained 12 ... 24 h at 5 ... 10 ° C, to complete the crystallization, followed by vacuum filtration, washing the precipitate with 0.1 ... 0.31 distilled water to neutral pH, drying at 105 ° C, for 3 hours and spraying, resulting in the bioactive acid complex. purified triterpenic acid, as Na or K or NH ₄ salt, with a content of at least 90%, RO 127000 Β1 consisting of: minimum 75% Na or Ksau ursolate of NH ₄ , 10 ... 15% oleanolate of Na 1 or K or NH ₄ and 4 ... 10% salts of Na, K or of NH ₄ of other triterpenic acids, consisting of one or more of the acids: hydroxyursolic, hydroxyoleanolic betulinic, 3 dehydrobetulinic. 5. Medicinal product for human and / or veterinary use, characterized in that 5 contains 20 ... 40% pharmaceutically active substance, represented by the complex described in claims 1 and 2, conditioned in the form of tablets, hard operculated capsules and / or ice cream - 7 soft nuts. 6. Medicinal product for human and / or veterinary use, characterized in that 9 contains, as a pharmaceutically active substance, the complex described in claims 1 and 2, conditioned in the form of gel or cream, in an amount of 0,1 ... 1%. 11 7. Medicinal product for human and / or veterinary use, characterized in that it contains, as a pharmaceutically active substance, the complex described in claims 1 and 2, 13 in an amount of 0.5 ... 1%, conditioned as suppositories. .