RO127000A2 - 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 present invention relates to a bioactive complex of triterpenic acids, as such or as Na, K or NH4 salts, to the process of obtaining Salvia species, and to medicinal products based thereon, applicable in wide-ranging therapeutic fields. for both human and veterinary use.

The bioactive complex realized according to the invention, has a continuum of triterpenic acids 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) it is known that they, although isomers, 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, the 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, the dependence 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 the substances influence their solubility in biological environments. Knowing this fact allows the prognosis of the behavior of medicinal substances from the point of view of bioavailability, in different regions of the gastrointestinal tract. The information provided by the bioavailability studies can determine the modification of the final product formulation and its manufacturing process as well as the extension of the therapeutic indications.

A main factor that can modify 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 directed for the improvement of their bioavailability.

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

a) the sources of plant raw materials:

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

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

b) the finished product that constitutes the main objective of the patented process:

- pure triterpenic acids, as such or as salts;

- mixture of triterpenic acids, triglycerides, sugars.etc. obtained mainly from fruit wastes.

Following are the patents of the invention and recent patent applications, which present processes for obtaining triterpenic acids (especially ursolic and / or oleanolic):

- Patent GR 10011738 entitled "Method of separation of ursolic and oleanolic acids from leaves of European O / ea, is a process at laboratory level, which envisages the extraction of dried leaves of European O / ea with ethyl alcohol 95 ^c , at temperature environment, for 15 days, 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. We present a method of analysis by thin layer chromatography, which helps to qualitatively evaluate the sediment purified with petroleum ether, as

being made of ursolic acid, and the petroleum ether extracts, the concentrate contains oleanolic acid.

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There is no mention of the quantitative evaluation of the ursolic and oleanolic acids thus separated, 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 oleanolic acid and / or olive acid", presents a method of preparing these acids as such or in the form of physiologically acceptable salts, using as a raw material the Olea europaea plant from which process the remaining material after the extraction of the olive oil. Hydrophilic organic solvents (alcohols, acetone) or lipophiles (hexane, chloroform, ethyl acetate) are used as extraction solvents, and the resulting extracts containing oleanolic acid or olive oil are purified by liquid extraction or chromatography on silica gel column.

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%.

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

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

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

- Patent RU 2108107, entitled: "Method for preparing a biologically active total of salts of triterpenic acids", presents a process for obtaining triterpenic acids of species of white fir (Abies sp.), In the form of a total of salts of Na of triterpenic acids, which provides for the extraction of plant raw material with petroleum ether mixture - 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 aqueous alkaline extract, with pH = 7 ... 8 is concentrated in vacuo, until a dry powder extract is obtained, which constitutes the total of triterpenic acids, as a salt of Nâ <

38 to 45% content.

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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 of 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, treating organic extract with alkaline agent (aqueous solution 2% NaOH), 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 organic solvents ( methyl-butyl ether; mixture: benzene-ethyl acetate; mixture: petroleum ether-ethyl acetate), 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 based on which the possibilities can be evaluated. 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.

EP patent 1161879] and US application 2002037882], present 2 laboratory and pilot procedures - which extraction of dried apple shells with acetone at reflux (5O ... 58 ° C),

patent provide

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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 presented 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 (H3PO4) at pH = 2 and with aqueous alkaline solutions (Na2CC> 3) at pH = 8-12, followed by extraction with acetone at Soxhlet and concentration of the extractive solution at sicc residue, which is the concentrate of ursolic and oleanolic acid 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.

- Patent CN 1358733 presents a process for the extraction of 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, concentration of the extract to the residue, which is washed with water, dried and the extract dry it is dissolved in ethyl alcohol and it is discolored, then concentrated and diluted with water, resulting in a precipitate; adjust the pH of the suspension to 2 ... 2.5, after which the precipitate is separated, washed with water at neutral pH, and dried under vacuum.

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

- CN1508149, presents a method of extraction of total triterpene acids from kudingcha llex with ethyl alcohol as solvent through a process of reflux distillation or microwave extraction, followed by a series of separation steps by ultrafiltration, nanofiltration, chromatography with resins macroporous and purification by discoloration with activated carbon and separation using ether or an organic acid, obtaining in / inal a total triterpenic acid concentration> 95%.

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- CN1453289, 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 acetone and recrystallization 3 times with ethanol or anhydrous methanol.

- W02005075614 (A1) - shows the extraction of the olive leaves with hexane, ethyl alcohol, followed by the concentration operations, separation on an extraction column in countercurrent supercritical carbon dioxide.

- CN1557831-presents a process of extraction of ursolic acid from rosemary through Soxhlet extraction, concentration, dissolution in ethanol, discoloration with coal, crystallization, filtration after 3-6 hours obtaining crystals of ursolic acid of about 90% purity.

- CN101318985- reports a method of obtaining ursolic acid from the leaves of the plant loquat with ethyl alcohol 95% as solvent by ultrasonication, followed by filtration through the nanofiltration micromembrane, recovery of ethanol and obtaining ursolic acid by mixing with honey, in the form of a cream ursolic acid content> 75%.

- CN101538271- presents a method of simultaneous extraction of picrinin and ursolic acid by extraction from the aqueous acidic solution of the leaves of the plant called ultrasound, the extract obtained is volatilized, the concentrate is alkalized and subjected to extraction with chloroform from which by separation picrinin and ursolic acid.

- CN101537029- relates a method of preparing the extract of the horn powder which is subjected to the hydrolysis with yeast; fermentation lasts 4-8 days, the extract obtained is subjected to ultrasonication, filtration and horn powder so prepared is subjected to extraction with 75-95% ethanol by ultrasonication, filtration, concentration, obtaining a solid in the form of scales containing ursolic acid.

- RO120950 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 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, resulting in a powder that represents the active substance that can be used when conditioning some medicinal products for internal use.

From the data presented above, regarding the state of the art in the field of obtaining triterpenic acids used as the drug substance, by extracting the different sources of plant raw materials, the following results

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- most processes related to obtaining pure ursolic and oleanolic acids, do not mention the purity of the substance and its evaluation by quantitative analytical methods (HPLC, GC and others).

- the processes that claim the production of triterpenic acids concentrates, based on a mixture of ursolic and oleanolic acid, although they present their quantitative evaluation by HPLC, have 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;

- in addition, it is known that fruit wastes have a low content of triterpenic acids (<0.5%) which negatively affects the profitability of the technologies at 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 amorphous structure, the amorphous state being thermodynamically unstable, it influences the stability of the structure of the drug substance;

- none of the presented processes has technological data similar to the process that is the subject of the present invention nor pharmacological data regarding the specific biological activity of the claimed triterpenic acid mixture, in order to evaluate the fields of therapeutic applicability;

- there are no patents and data in the specialized 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 the drug 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, antirriicrobial.

Somova L.l. et al. highlights the cardiovascular, antihypertensive, antihyperlipidemic, antiaterosclerotic and antioxidant activity of oleanolic acids isolated from Olea europea.

And Kuttan G. et al. mentions the ursolic, oleanolic and glycirrhizic immunomodulatory activity. >

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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.

US Patent Application 10/488682] published with US no. 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 compounds of the terpenes class (mono-, di-, tri-, sequiterpene).

Patent application JP 05-211065] published under No. 07-048260 / 1995 presents a bioproduct that can be used as a blood cell growth agent (erythrocytes) based on ursolic acid, administered in daily doses of 1-1000 mg and having a prolongation effect in the radiotherapy treatment of different forms of cancer.

Patent applications EP 1495754 A2 published on 12.01.2005 and EP 0943620 A2 published on 22.09.1999, present a method and composition based on betulinic acid derivatives applicable for the prevention or inhibition of malignancies growth. There is presented a method of obtaining betulinic acid from the bark of the plant Ziziphus mauritiana (Rhamnacaee fam.), "In vitro studies on the cytotoxicity of betulinic acid against different cancer cell lines (melanoma, fibrosarcoma, breast and colon cancer, epidermoid carcinoma). and results of "in vivo" mouse experiments, which confirm the quality of betulinic acid as an antineoplastic agent.

U.S. Patent No. 5985924 discloses a novel low-toxicity metastasis suppressor, based on ursolic acid, as such or as salts, which may be administered to patients by bone or injectable, in the post-operative treatment of different forms of cancer.

The patent application JP 63166195 published no. 02017121 A / 1990 presents an medicinal product for external use, which can be used to prevent the cancer of epithelial cells of the skin, based on ursolic or oleanolic acid, as such or in combination.

The patent application JP 09-199323 published with the PG No. 11-029467 / 1999 presents a product with inhibitory effects on the protease, based on ursolic acid or its salts, applicable for the treatment or prevention of various skin dysfunctions (dermatitis). and conditioned in the form of ointment, cream, lotion and emulsion. it mentions the dermatoprotective effects of the product: antioxidants, hydrates and skin bleaching.

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

60 ... 70%.

From the present data on the state of the art in the field of pharmacology of triterpenic acids, the following results:

- all pharmacological studies related to the specific activity of these acids have been performed separately for each acid, with a multitude of pharmacological effects that, although largely similar, are not identical;

- none of the mentioned studies presents pharmacological tests performed on a mixture of triterpenic acids, in order to highlight the synergism and potentiate the pharmacological effects, which may be superior to those of each component;

- the preclinical or clinical pharmacological experiments presented in the mentioned studies were not performed in comparison with young, adult or aged organisms, although it is known that the biochemical and hematological parameters differ significantly depending on age, which influences the therapeutic efficacy of the drug used.

The technical problem solved by the present invention consisted in the elaboration of process conditions by which a bioactive complex with a specific design for a certain type of crystalline structure is obtained, which favors its solubility, resulting in an improvement of the bioavailability of the active substance. This fact in turn allows to increase the efficacy of the product and to a potential expansion of the therapeutic area of the drug.

The bioactive complex of triterpenic acids, consists in the fact that it has a content of 100% triterpenic acids, consisting of at least 80% ursolic acid and max. 20% oleanolic, hydroxyursolic, hydroxyoleanolic, betulinic, dehydrobetulinic acid, obtained in the form of state acids. free or as salts of Na, K or NH ₄ by extraction from different species of Salvia, having the following pharmacological effects:

- activity of bioregulation of protein metabolism, carbohydrate and lipid, especially on aging organisms, with applications in preventive and curative therapy of metabolic and cardiovascular diseases with atherosclerotic etiology, especially in geriatrics; //

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- immunoprotective and hepatoprotective activity, to counteract the immunosuppressive and hepatotoxic adverse effects, which occurred after treatment with cytostatic glucocorticosteroids;

- activity to delay the evolution of tumors;

- antioxidant activity;

- local anti-inflammatory activity.

and the process for obtaining the bioactive complex of triterpenic acids consisted in the fact 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 ^c , for

8 .. .24h, at 15 ... 80 ° C, resulting in the extraction solution which, for the purpose of purification, is passed on a chromatographic column with activated carbon granulated with acid pH, then concentrated under vacuum at 35 ... 40 ° C up to 1/10 ... 1/20 of the initial volume obtaining a suspension that is maintained for 12 .... 24 h at

5 .. ..10 ° C, then filtered 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, is 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 or 90 ^c 95 ... 95% acetone; the solution obtained is purified by adsorption on activated carbon with acid pH, added in the ratio of 0.1 to 0.3% m / v, filtered, concentrated under vacuum at 35 ... 40 ° C to 1 /10...1/20 from the initial volume, and the resulting suspension was left 12 ... 24 h at 5 .... 10 ° C, filtered under vacuum, the precipitate obtained was washed with 0.3 .... 0.5 I distilled water, up to neutral pH, is dried at 105 ° C for a minimum of 3 hours, sprayed, obtaining the sprayed product with a concentration of at least 90% triterpenic acid subjected to crystallization from ethyl alcohol, acetone or ethyl acetate by dissolution under reflux under vacuum, 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 a bath of ice quickly, under stirring, is filtered immediately under vacuum and dried at 90-110 ° C in hot air stream, obtaining the bioactive complex of triterpenic acids 100% of which ursolic acid min. 80% and m shaft. 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? tablet ^ -2010-00578-0 5 -07- 2010

or soft and / or soft gelatin capsules, gel, cream or ointment, external use spray or spray, suppositories or ova.

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

- the preclinical pharmacological studies "in vivo", performed on the bioactive complex of triterpenic acids, have shown that it has a superior pharmacological activity than that of each separate component, due to synergistic effects of mutual potentiation, used in obtaining new medicinal products;

- expects an increase of 10 ... 15% of the extraction efficiency;

- there is a high degree of selectivity of the extraction process;

- 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 its content or components;]

- the modeling phase 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 the dissolution process, but 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, presents a content of 100% standardized, having a marked and diversified pharmacological activity, exerted at the level of some 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 as well; capsules, tablets, injectable solutions, oral and for external use; eye drops, spray, gel, cream, ointment, suppositories, ova, intended for new therapeutic applications, compared to those currently known.

The process according to the invention consists in that the dried and pulverized vegetable raw material, with 2 ... 4% triterpenic acids, consisting of Salvia spec / es herb, is extracted by dynamic maceration in the ratio 1/15 ... 1 / 30 m / v, plant mass / soil, no extraction solvents represented by the mixtures constituted

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structured water "I" with 85 ... 95% acetone or 85 ... 95% ethyl alcohol 95 ^c , for 8 ... 24 h, at 15 ... 80 ° C, resulting in the extractive solution, which in the purpose of the purification is passed on a chromatographic column with activated carbon granulated with acid pH, then concentrated in vacuo, at a temperature of 35 ... 40 ° C, up to 1/10 .... 1/20 of the initial volume, obtaining -is a suspension which is kept for 12 ... 24 h at 5 ... 10 ° C, then is filtered under vacuum, the substance obtained after drying at 105 ° C, for 3 h, constituting the bioactive complex of triterpenic acids gross 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 one of the solvents represented by the mixtures consisting of: 5 ... 10% structured water "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 0.1 ... 0.3% m / v, filtered, concentrated in vacuo at 35 ... 40 ° C to 1/10 ... 1/20 of the initial volume, the resulting suspension is left 12 ... 24h at 5 ... 10 ° C, filtered under vacuum, the precipitate obtained is washed with 0.3 ... 0.5 I distilled water , to neutral pH, is dried at 105 ° C for a minimum of 3 hours and sprayed, obtaining the sprayed product with a concentration of at least 80% triterpenic acids which is subjected to crystallization from ethyl alcohol, acetone or ethyl acetate by dissolution at under reflux under vacuum, with the rising of the vapor temperature to 50 ° .... 60 ° C, concentration under vacuum up to 1/10 -1/20 of the initial volume, the crystalline suspension obtained cools rapidly on the ice bath, with stirring, it is filtered immediately under vacuum and dried at 90-110 ° C in hot air stream, obtaining the bioactive complex of 100% triterpenic acids of which ursolic acid min. 80% and max. 20% mixture of oleanolic, hydroxyursolic, hydroxyoleanolic, betulinic, dehydrobetulinic acid ...

In order to obtain the complex of triterpenic acids in the form of salts of Na, K, or NH _4, the sprayed product with a concentration of at least 80% triterpenic acids dissolves in the ratio 1/100 ... 1/200 m / v in the solvent mixture with Alkaline pH: 90 ... 95% ethyl alcohol 95 ^c with 5 ... 10% structured "S" water, which also contains 1% NaOH or 1% KOH or 5% NH ₄ OH 25%, followed by the concentration of the resulting solution and obtaining the bioactive complex of triterpenic acids as Na, K or NH ₄ salt, which is subjected to crystallization from ethyl alcohol, acetone or ethyl acetate by dissolution under vacuum under vacuum, 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, immediately filtered under vacuum and dried at 90-110 ° C. in hot air stream, obtaining the bioactive complex of triterpenic acids as Na salt, K s with a minimum content of 9 0%, evaluated and a phase yield of 95%. f \ Γ '· ¹ '; B1OTEHNOS12 0

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CV 2 0 1 0 - 0 0 5 7 8 -0 5 -07- 2010 The following are examples of embodiments of the invention, which refer to the process of obtaining the bioactive complex of triterpenic acids and to the conditioned medicinal products.

The embodiment example no. 1 - Variant A. Obtaining the bioactive complex of triterpenic acids at the industrial scale of the grass Salvia species (Salvia) kg dry and pulverized vegetable raw material, consisting of the grass Salvia species (Salvia officinalis, Salvia lavandulifolia, Salvia triloba,), with a content of 3 ... 4% triterpenic acids, extracted with 500 I extraction solvent consisting of: 90 volumes (450 I) acetone + 10 volumes (50 I) structured water "I", with pH = 1.8 ... 2 , 6, by dynamic maceration, for 12 hours, at ambient temperature (15 ... 30 ° C), after which a minimum of 400 I extraction solution is collected which is passed on a chromatographic column with 2 kg acid granulated activated carbon, resulting 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, after which filter under vacuum, and the precipitate on the filter is dried at 105 ° C for 3 h and spray. A minimum of 1200 g of light greenish yellow powder is obtained, which constitutes the bioactive complex of crude triterpenic acids, with a content of at least 70%.

For the purification of the product, the solvent mixture with acid pH consisting of 90 volumes (108 I) ethyl alcohol 95 ^c +10 volumes (12 I) structured water "I", with pH = 1.8 ... 2.6, is used. to which is added for dissolution 1200 g powder consisting of the bioactive complex of crude triterpenic acids (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 kept 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 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, immediately filtered under vacuum and dried at 90-110 ° C in hot air stream, obtaining 600 g white crystalline substance representing the bioactive complex of triterpenes 100% of which ur acid ask for min. 80% and max. 20% mixture / of cy2 C rings - 0 5 7 8 -o -07- 2010 triterpenic acids, consisting of one or more of the acids: hydroxyursolic oleanolic acid, hydroxyoleanolic acid, betulinic, dehydrobetulinic acid, representing a final yield of at least 50 % related to the triterpenic acid content of the processed vegetable 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 500 I extraction solvent, consisting of: 90 volumes (450 I) ethyl alcohol 95 ^c +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 400 I extractive solution is collected, which is passed on a chromatographic column with 2 kg acid granulated activated carbon, resulting in a purified extractive solution, which is concentrated in vacuo , at a temperature of 35 ... 40 ° C, up to a volume of 40 I, giving a suspension which is kept for 24 hours at a temperature of 5 ... 10 ° C, after which it is filtered under vacuum and the precipitate on The filter was dried at 105 ° C for 3 hours and sprayed. At least 1200 g of yellow powder is obtained, slightly greenish, which constitutes the complex of crude triterpenic acids, with a content of minimum 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. add, for dissolution, 1200 g of powder consisting of the crude triterpenic acid complex (calculated for 1 g of substance - 100 ml of 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 5 ... 10 ° C to complete the crystallization, after which it is filtered under vacuum, the precipitate is washed with 0.5 I distilled water, until neutral pH, dried at 105 ° C, for 3 hours it is sprayed. the complex of triterpenic acids purified with a content of at least 80% which 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 quickly, with stirring, immediately filtered under vacuum and dried at 90-110 ° C in hot air stream, yielding 610 g of the whitish white substance represents the bioactive complex of 100% triterpenic acids of which ursoleic acid

/

_λ -2910-00578-0 5 -07- 2010 max. 19% mixture of other triterpenic acids, consisting of one or more of the acids: hydroxyursolic oleanolic acid, hydroxyoleanolic, betulinic, dehydrobetulinic acid, which represents a final yield of at least 50% related to the triterpenic acid content of the processed plant raw material.

The embodiment example no. 2. Obtaining the bioactive complex of triterpenic acids in the form of Na salt

200 g purified bioactive complex powder of triterpenic acids, with a content of at least 80%, obtained according to the working method described in the embodiment no. 1 is dissolved in 20 I (1/100 m / v ratio) solvent mixture consisting of: 90 volumes (18 I) ethyl alcohol 95 ^c + 10 volumes (2 I) structured water "S" (pH = 10 ... 12), containing 1% NaOH, by 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 microcrystalline suspension, which is maintained for 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 L 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 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, immediately filtered under vacuum and dried at 90-110 ° C in hot air stream, obtaining 180 g of the white crystalline substance representing the complex bioactive triterpene acids in the form of Na salt of ursolic, oleanolic acids and other triterpenic acids consisting of one or more of the acids: hydroxyursolic, hydroxyoleanolic, betulinic, dehydrobetulinic, with a content of min 90%, which represents a yield of min. phase of 95%.

The embodiment example no. 3. Obtaining the bioactive complex of triterpenic acids in the form of K salt

200 g bioactive complex purified of triterpenic acids, with a minimum content

80%, obtained according to the working mode described in the embodiment no. 1, dissolves in 20 I solvent mixture consisting of: 90 volumes (18 I) ethyl alcohol 95 ^c + 10 volumes (2 I) structured water "S" (pH = 10 ... 12), containing 1% KOH , by heating under reflux (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 for 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 I neutral distilled water, dried at 105 ° C for 3 h, sprayed and

• BOTEHNOS · (Χ-2 0 1 0-00570-0 5 -07 “2010 of a 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, immediately filtered under vacuum and dried at 90-110 ° C in hot air stream , obtaining 180 g of the white crystalline substance 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, dehydrobethulin, with a content of min. 90%, which represents a phase yield of 95%.

The embodiment example no. 4. Obtaining the bioactive complex of triterpenic acids in the form of NH4 salt

200 g bioactive complex of purified triterpenic acids, with a content of at least 80%, obtained according to the working method described in the embodiment no. 1, dissolves in 20 L solvent mixture consisting of: 90 volumes (18 I) ethyl alcohol 95 ^c + 10 volumes (2 I) structured water "S" (pH = 10 ... 12), containing 5% NH4OH 25% by heating under reflux (80 ° C); The resulting clear solution is concentrated in vacuo at

40 ... 45 ° C, up to 1/4 of the initial volume (5 I), resulting in a microcrystalline suspension, which is maintained 24 h, at 5 ... 10 ° C, in order to complete the crystallization, after which filter under vacuum, the precipitate on the filter is washed with minimum 0.1 L distilled water to neutral pH, dried at 105 ° C for 3 h, sprayed and subjected to crystallization from organic solvents, acetate ethyl by dissolution under 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 cools rapidly on the ice bath , with stirring, it is filtered immediately in vacuo and dried at 90-110 ° C in hot air stream, obtaining 180 g of white crystalline substance representing the bioactive complex of triterpenic acids in the form of NH ₄ salt of ursolic acids, oleanolic and other triterpenic acids consisting of one or more of the acids: hi droxiursolic, hydroxyoleanolic, betulinic, dehydrobetulinic, with a content of at least 90%, which represents a phase yield of 95%.

The embodiment example no. 5 - Variant A. Obtaining the bioactive complex of triterpenic acids on the industrial scale from the grass Salvia species (Salvia) kg dry and pulverized vegetable raw material, consisting of the grass 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 J45îȚJ) = acetone + 10 volumes (50 I) structured water "I", with pH = 1.8 ... 2.6, Prix / macerating

O 1 O - 0 O 5 7 8 'O 5 -07- 2010

dynamic, 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 2 kg of acidic granulated activated carbon, resulting in a purified extractive solution, which are concentrated in vacuo at 35 ... 40 ° C, up to the volume of 40 I, obtaining a suspension which is kept for 24 hours at 5 ... 10 ° C, after which it is filtered under vacuum, and the precipitate on the filter was dried at 105 ° C for 3 h and sprayed. A minimum of 1200 g of light greenish yellow powder is obtained, which constitutes the bioactive complex of crude triterpenic acids, with a content of at least 70%.

For the purification of the product, the mixture of solvents with acid pH consisting of 90 volumes (108 I) ethyl alcohol 95 ^c + 10 volumes (12 I) structured water "I", with pH = 1.8 ... 2.6, is used. to which is added for dissolution 1200 g powder consisting of the bioactive complex of crude triterpenic acids (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 kept 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 crystallization from organic solvents, ethyl alcohol by dissolution at reflux under vacuum, with raising the vapor temperature to 50 ° .... 60 ° C, concentration under vacuum until at 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, obtaining 550 g white crystalline substance representing the bioactive complex of triterpenic acids 100% of which ac ursolic id min.80% and max. 20% mixture of other triterpenic acids, consisting of one or more of the acids: hydroxyursolic oleanolic acid, hydroxyoleanolic, betulinic, dehydrobetulinic 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 500 I extraction solvent, consisting of: 90 volumes (450 I) ethyl alcohol 95 ^c + 10 volumes (5 structured "I" (pH = 1.8 ... 2.6), by dynamic maceration, for 12 hours, at tşfţ;

i

cv 2 O 1 0 - OO 5 7 8 - O 5 -07- 2010 ambient (15 ... 30 ° C), after which a minimum of 400 I extractive solution is collected, which is passed on a chromatographic column with 2 kg of activated carbon acid granulate, resulting in a purified extractive solution, which is concentrated in vacuo, at a temperature of 35 ... 40 ° C, up to a volume of 40 l, giving a suspension which is maintained for 24 hours at a temperature of 5 ... 10 ° C, then filtered under vacuum, and the precipitate on the filter was dried at 105 ° C for 3 h and sprayed. At least 1200 g of yellow powder is obtained, slightly greenish, which constitutes the complex of crude triterpenic acids, with a content of minimum 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. add, for dissolution, 1200 g of powder consisting of the crude triterpenic acid complex (calculated for 1 g of substance - 100 ml of 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 5 ... 10 ° C to complete the crystallization, after which it is filtered under vacuum, the precipitate is washed with 0.5 I distilled water, until neutral pH, dried at 105 ° C, for 3 hours it is sprayed. the complex of triterpenic acids purified with a content of at least 80% which is subjected to crystallization from organic solvents, ethyl alcohol by dissolution at reflux under vacuum, with raising the vapor temperature to 50 ° .... 60 ° C, concentration under vacuum until at 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 560 g white crystalline substance representing the bioactive complex d is triterpenic acids 100% of which ursolic acid min.81% and max. 19% mixture of other triterpenic acids, consisting of one or more of the 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.

The embodiment example no. 6 - Variant A. Obtaining the bioactive complex of triterpenic acids at the industrial scale of the grass Salvia species (Salvia) kg dry and pulverized vegetable raw material, consisting of the grass Salvia species (Salvia officinalis, Salvia lavandulifolia, Salvia triloba, etc.), with 3χ4 % triterpenic acids, extracted with 500 l extraction solvent constituted

^ -2 0 1 0 - 0 0 5 7 9 -Q 5 -07- 2010 (450 I) acetone + 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 400 l of extractive solution is collected which is passed on a chromatographic column with 2 kg of acid granulated activated carbon, resulting in an extractive solution purified, which is concentrated in vacuo at 35 ... 40 ° C, up to the volume of 40 I, having obtained a suspension which is kept for 24 hours at 5 ... 10 ° C, after which it is filtered in vacuo , and the precipitate on the filter is dried at 105 ° C for 3 hours and sprayed. A minimum of 1200 g of light greenish yellow powder is obtained, which constitutes the bioactive complex of crude triterpenic acids, with a content of at least 70%.

For the purification of the product, the mixture of solvents with acid pH consisting of 90 volumes (108 I) ethyl alcohol 95 ^c + 10 volumes (12 I) structured water "I", with pH = 1.8 ... 2.6, is used. to which is added for dissolution 1200 g powder consisting of the bioactive complex of crude triterpenic acids (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 kept 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, 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, immediately filtered under vacuum and dried at 90-110 ° C in hot air stream, obtaining

580 g white crystalline substance representing the bioactive complex of triterpenic acids 100% of which ursolic acid min. 80% and max. 20% mixture of other triterpenic acids, consisting of one or more of the 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.

Variant B. 30 kg plant raw material, dried and sprayed, consisting of grass Salvia species (Salvia officinalis, Saivia lavandulifolia, Salvia triloba, Salvia sclarea etc.), with a content of 3-4% triterpenic acids, extracted with 500 I ground

extraction, consisting of: 90 volumes (450 l) ethyl alcohol 95 ^c + 10 volumes

.. - λ

The water: biotechnos 4

... to ι _f 49 / il ../ Λ Z—

Λ-2 Ο 1 Ο - Ο Ο 5 7 8 - Ο 5 -07- 2010 structured "I" (ρΗ = 1.8 ... 2.6), by 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 2 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, giving 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 yellow powder is obtained, slightly greenish, which constitutes the complex of crude triterpenic acids, with a content of minimum 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. add, for dissolution, 1200 g of powder consisting of the crude triterpenic acid complex (calculated for 1 g of substance - 100 ml of 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 5 ... 10 ° C to complete the crystallization, after which it is filtered under vacuum, the precipitate is washed with 0.5 I distilled water, until neutral pH, dried at 105 ° C, for 3 hours it is sprayed. the complex of triterpenic acids purified with a content of minimum 80% which is subjected to a crystallization of acetone 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, it is filtered immediately under vacuum and dried at 90-110 ° C in hot air stream, yielding 590 g of the white crystalline substance. represents the bioactive complex of 100% triterpenic acids d in which ursolic acid min.81% and max. 19% mixture of other triterpenic acids, consisting of one or more of the 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.

(^ -2 0 1 0 - 0 0 5 7 0 -

Ο 5 -Ο? - 2010

EXAMPLES OF CONDITIONING OF THE BIOACTIVE COMPLEX OF TRITERPENIC ACIDS UNDER THE FORM OF MEDICINAL PRODUCTS based on the results of the pharmacological tests carried out, the bioactive complex of triterpenic acids as such, or in the form of salts of Na, K or NH4, 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 therapeutics. The medicinal products described below, according to the invention, are administered in human or veterinary therapy, by injection, oral or local, for 1 day 24 months, depending on the evolution of the disease, in daily doses of 10-1000 mg / kg. m.c., divided into 16 sockets. The examples of formulations presented below do not exclude the possibility of making other conditioning formulas, given the diversity and multitude of excipients known and used in the pharmaceutical industry.

The embodiment example no. 7

MEDICINAL PRODUCT CONDITIONED IN CAPSUMS CONTAINING 100 mg BIOACTIVE COMPLEX OF TRITERPENIC ACIDS / CAPSULA

Conditioner / capsule formula:

- bioactive complex of triterpenic acids -100% 105 mg

- lactose monohydrate 150 mg

- sodium starch glycol9.0 mg

- polyvinylpyrrolidone K30 6.5mg

- sodium lauryl sulfate 2.0mg

- aerosil 1,47mg

- talc 7.25mg

- magnesium stearate 2.9mg

Method of preparation: The bioactive complex of triterpenic acids is located with the excipients: lactose monohydrate and aerosil on a size II screen. Over the mixture obtained, pour a solution obtained by dissolving the polyvinylpyrrolidone K 30 in ethyl alcohol, and homogenize. Granules are obtained which are dried at a temperature of

35 ... 45 ° C and then reduced to the size conferred by sieve V; add sodium starch glycolate, sodium lauryl sulphate, 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 anti-ulcer, <r ~

\\ / 1 // <BIOTEHNOS 2Ls.a.

') Pharmacological effects: antitumorigenic, immunoprotective, hepatoprptector hypoglycemic antiinflamatoZ t0nMarit ^{AA' v} u

vascular, especially at coronary level, with therapeutic uses in the following diseases:

neoplasm, depletion of the immune system, acute and chronic hepatitis, liver cirrhosis, ulcer disease, diabetes, cardiovascular disorders with atherosclerotic etiology, degeneration of vascular, tissue and dermal collagen: psoriasis, neurodermitis, radiodermitis, varicose veins syndrome and different , retinal vascular diseases, applicable also in geriatrics to prevent and slow down aging processes.

The embodiment example no. 8

MEDICINAL PRODUCT CONDITIONED IN THE TABLE OF CONTENTS 100 mg BIOACTIVE COMPLEX OF TRITERPENIC ACIDS / TABLE

Conditioner / tablet formula: - bioactive complex of triterpenic acids -100% - lactose monohydrate 105.00 mg 81.20 mg - microcrystalline cellulose 74.00 mg - corn starch - sodium starch glycolate - polividona - magnesium stearate - aerosil 16.20 mg 5.00 mg 5.60 mg 2.00 mg 1.00 mg - talc 5.00 mg

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 the various manifestations of chronic venous insufficiency, retinal vascular diseases, also applicable in geriatrics for the prevention and) discontinuation of aging processes.

r

^ - 2 9 1 0-OQ5 7B- Ο 5 -07- 2010

The embodiment example no. 9

MEDICAMENTAL PRODUCT CONDITIONED IN THE FORM OF GEL - DERMATOLOGICAL USE CONTAINING 1% BIOACTIVE COMPLEX OF TRITERPENIC ACIDS - Salt of Na Conditioning formula:

bioactive complex of triterpenic acids - minimum 90% Na salt 10.5 g Carbopol 940 20 g glycerin 60 g Sodium hydroxide solution 15% 10g propylene glycol 60 g methyl p-hydroxybenzoate Ig 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 triterpenic acid bioactive complex, 60 g propylene glycol and 10 g 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 , articular and dermal, anti-inflammatory, anti-hyaluroriidazine and anti-elastic, 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 conditions, 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.

The embodiment example no. 10

MEDICINAL PRODUCT CONDITIONED IN THE FORM OF CREAM

CONTAINING 1% BIOACTIVE COMPLEX OF TRITERPENIC ACIDS - Na salt

Conditioning formula:

- bioactive complex of triterpenic acids - Na salt minimum 90% 10J £ Îj

r '

W> BIOTEHNO5 · \

SADDLE. 'That in./

Ή

^ - 2 0 1 0 - 0 0 5 7 8 0 5 -07- 2010 cetyl alcohol 200 g white Vaseline 200 g glycerin 120 g Tween 80 70 g methyl p-hydroxybenzoate ig propyl p-hydroxybenzoate 2g structured water "I" (pH = 1.8-2.7) 1000 g

Method of preparation: 10.5 g of bioactive triterpenic acid - Na salt is dissolved in 120 g of glycerin, and the resulting solution is incorporated into the ointment mass, obtained by melting at 60 ° C of cetyl alcohol and vaseline, followed by emulsification by adding 70 g Tween 80 and at least 400 g structured water "I" in which the preservatives provided in the formula have been dissolved previously. The mass of ointment is stirred for emulsification and homogenization, then packaged in tubes of 50 g.

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

The embodiment example no. 11

MEDICAMENTAL PRODUCT CONDITIONED IN THE FORM OF SUPPOSERS CONTAINING 1% BIOACTIVE COMPLEX OF TRITERPENIC ACIDS

Conditioning formula: - bioactive complex of triterpenic acids 100% 1.05 g - polyethylene glycol 400 5g - methyl para-hydroxybenzoate 0.1 g - propyl para-hydroxybenzoate 0.03 g - Supplementation (mixture of semisynthesis triglycerides) ad. 100 g Preparation: dissolve the bioactive complex of triterpenic acids in

polyethylene glycol, by heating to 80 ° C. The solution obtained is cooled to 40 ° C

'II · BIOTEHNOS i ₍ \ * 24.A. 'I,

Z '/ 2.

^ 2010-00570-Ο 5 ~ 07 ~ 2010

over the Supocir table which was previously heated to 4 ° C. Add the mixture of preservatives, mix and pour into the suppositories that weigh every 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 effects: immunoprotective, anti-inflammatory, trophic and regenerative of collagen. vascular, tissue and joint, with uses in the following diseases: immunosuppressive, especially hepatotoxic, occurring after treatment with glucocorticosteroids, cytostatic, inflammatory rheumatic diseases, hemorrhoidal syndrome.

Claims (8)

1. Bioactive complex of triterpenic acids, characterized in that it has a crystalline structure, improved solubility and a continuum of 100% acids, made up of min 80% ursolic acid and max. 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 , by extraction of the plant Salvia species having the following pharmacological effects: - bioregulation activity of protein metabolism, carbohydrate and lipid metabolism, especially on aging organisms, with applications in preventive and curative therapy of metabolic and cardiovascular diseases with atherosclerotic etiology; - immunoprotective and hepatoprotective activity, to counteract the immunosuppressive and hepatotoxic adverse effects, which occurred after treatment with cytostatic glucocorticosteroids; - antioxidant activity; - local anti-inflammatory activity. 2. The bioactive complex of triterpene acids according to claim 1, characterized in that, in order to increase bioavailability and solubility of the product in each of the conditioning solvent is obtained and used in the form of salts of Na, K or NH ₄ , of minimum 90% purity, consisting of: minimum 75% Na or K or NH4 ursolate, 10 ... 15% Na or K or NH or NH ₄ and 4 ... 10% Na, K or NH ₄ of other triterpenic acids consisting of one or more of the 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 ^c , for 8 ... 24h, at 15 ... 80 ° C, resulting in the extraction solution which, for the purpose of purification, is passed on a column chromatography with activated carbon granulated with acid pH, then concentrated under vacuum at 35 ... 40 ° C up to 1/10 ... 1/20 of the initial volume giving a suspension which is maintained 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%, where / n CV 2 Ο 1 Ο - Ο 0 5 7 8 - Ο 5 -07- 2010 purification, dissolves at reflux- 80 ° C in the ratio 1/100 ... 1/200 m / v in one of the solvents represented by the mixtures formed of: 5 ... 10% structured water "l" or "S" with 90 ... 95% ethyl alcohol 95 ^c or 90 ... 95% acetone; the obtained solution is purified by adsorption on activated carbon with acid pH, added in the ratio of 0.1 to 0.3% m / v, filtered, concentrated in vacuo at 35 .... 40 ° C until 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 I distilled water, up to neutral pH, is dried at 105 ° C for a minimum of 3 hours, sprayed, obtaining the purified product with a concentration of at least 90% triterpenic acid which is crystallized from ethyl alcohol, acetone or 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 the bath of ice quickly, under stirring, it is filtered immediately in vacuo and dried at 90-110 ° C in hot air stream, obtaining the bioactive complex of triterpenic acids 100% of which ursolic acid min. 80% and m shaft. 20% mixture of oleanolic, hydroxyursolic, hydroxyoleanolic, betulinic, dehydrobetulinic acid. 4. Process according to claim 3, characterized in that in order to obtain the bioactive complex of triterpenic acids in the form of Na, K and NH ₄ salts, as described in claim 2, the bioactive complex of triterpenic acids crystallized with a content of at least 90. % dissolves under 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 ^c with 5 ... 10% structured water "S", 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 to 35. .40 ° C to 1/5 ... 1/10 of the initial volume, the resulting suspension is maintained 12 ... 24h at 5 ... 10 ° C to complete crystallization, followed by vacuum filtration, washing the precipitate with 0 , 1 ... 0,3 I distilled water to neutral pH, drying at 105 ° C, for 3 hours and spraying, resulting in the purified triterpenic acid bioactive complex, as Na or K or NH ₄ salt, with a content of at least 90% consisting of: minimum 75% Na or K or NH ₄ ursolate, 10 ... 15% Na or K or NH or ₄ or 10 ... 10% Na salts K or NH ₄ of other triterpenic acids consisting of one or more of the acids: hydroxyursolic, hydroxyoleanolic betulinic, dehydrobetulinic. The product contains described medicinal products for human and / or veterinary use, characterized in that, as a pharmaceutically active substance, the bioactive acid complex Jriter in claims 1 and 2, conditioned in the form of capsules o Λ / \ / ^ - 2010-00570-O 5 -07- 2010 and / or soft gelatin containing between 50 ... 100 mg bioactive complex of triterpenic acids / capsule and administered in doses of 3 ... 6 capsules / day, for 1 ... 24 months, having the following pharmacological effects: antiaterosclerotic, vascular toning, especially at the coronary level, hypoglycemic, hypolipidemic, hepatoprotective, immunoprotective, antioxidant, anti-inflammatory, with therapeutic uses in the following diseases: cardiovascular disorders with atherosclerotic etiology, diabetes degeneration of vascular, tissue and dermal collagen, varicose syndrome and various manifestations of chronic venous insufficiency, retinal vascular disease. 6. Medicinal product characterized in that it contains, as a pharmaceutically active substance, the bioactive complex of triterpenic acids described in claims 1 and 2, conditioned in the form of tablets containing between 50 ... 100 mg bioactive complex of triterpenic acids / tablet and which are it is administered in doses of 3 ... 6 tablets / day, during 1 ... 24 months, having the following pharmacological effects: antiaterosclerotic, vascular toning, especially at the coronary level, hypoglycemic, hypolipidemic, hepatoprotective, immunoprotective, antioxidant, anti-inflammatory, with therapeutic uses in the following diseases: cardiovascular disorders with atherosclerotic etiology, diabetes degeneration of vascular, tissue and dermal collagen, varicose syndrome and various manifestations of chronic venous insufficiency, retinal vascular disease. 7. A medicinal product for human and / or veterinary use, characterized in that it contains, as a pharmaceutically active substance, the bioactive complex of triterpenic acids described in claims 1 and 2, conditioned in the form of gel or cream containing 0.1 ... 1. % bioactive complex of triterpenic acids, as such or in the form of salts that are administered in the dose of 10 ... 30 mg / day, for 1 ... 6 months, having the following pharmacological effects: trophic and regenerative of vascular collagen, tissue and dermal, anti-inflammatory, antioxidant, antihyaluronidase and anti-elastic, with therapeutic uses in the following diseases: degeneration of dermal, vascular, tissue collagen such as, inflammatory and pruritic dermatoses, varicose and post-thrombotic syndrome, chronic venous insufficiency, chronic venous insufficiency, fragility inflammatory. 8. A medicinal product for human and / or veterinary use, characterized in that it contains, as a pharmaceutically active substance, the bioactive triterpenic acid complex described in claims 1 and 2, conditioned in the form of suppositories containing 0.5 ... 1% bioactive complex of triterpenic acids, administered in doses of 50 ... 100 mg / day for 1 to 6 months, having the following pharmacological effects: immunoprotective, anti-inflammatory, antioxidant, trophic and regenerative of collagenesis xice \ .BIOTEHNOS: vascular, with uses in the following immunosuppressive states, especially h ocl o 1 o - o O 5 7 8 - O 5 Ό7-2010, occurring after treatment with glucocorticosteroids and cytostatics, inflammatory processes, hemorrhoidal syndrome.