New milk thistle extract, method for preparing and using it. RU patent 2489161.

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a method for preparing a milk thistle extract having a higher degree of silymarin release. A method for preparing a milk thistle extract, wherein the extract containing 15-85 wt %, particularly 30-65 wt % of silymarin is treated with anhydrous C1-C4 alcohol, optionally filtered and concentrated, then dried and optionally ground, wherein a degree of silymarin release is 80% or higher (versions). The milk thistle extract for treating and preventing liver and gallbladder dysfunctions. A pharmaceutical composition for treating and preventing liver and gallbladder dysfunctions.

EFFECT: extract is characterised by a higher degree of silymarin release and effective for treating and preventing the liver and gallbladder dysfunctions.

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The present invention relates to a method for the extract of the fruit milk Thistle, in particular, the composition, containing , which has a higher degree of release and improved absorbability, and application, of the composition, in particular for the treatment and prevention of liver diseases.

Milk Thistle (Silybum marianum or Carduus marianus) is a plant that bred, in particular, in the South-Western and Central Europe (Austria, Hungary) and that in Eurasia, North America, South America and Australia. The area of distribution is also found in China.

Well known for the effectiveness of drugs derived from milk Thistle (from seeds and fruits), prevention and treatment of various disorders of the liver and gallbladder. Medicine consists of ripe fruits, of which removed the flesh, with a minimum content of silymarin is equal to 1.5% (Pharmacopoea Europaea (Ph. Eur.), 2007). Tincture (usually alcohol extracts of medicinal substance) of milk Thistle known since ancient times. Especially widely used dedicated silymarin (see, for example, DE 1923982, DE 1767666 (Madaus)).

Silymarin is a mixture of , namely, . Silymarin was first isolated from plants in the 1960s (Dissertation, Janiak Bernhard, June 1960, Berlin University of Applied Sciences (DE 2020407), Pelter A., Hänsel R., Tetrahedron Letters, 25, (1968)).

Silymarin consists of a mixture of complexes I-IV; more specifically, its main components are four : silybin (silibinin) (silymarin I), (silymarin II), (silymarin III), as well as (silymarin IV). In these Taxifolin is connected with alcohol.

Other well-known secondary components are , 3-, (), , , and .

To extract use the fruits of the milk Thistle. Such extracts of milk Thistle and methods of their obtaining are described in the level of equipment, for example, DE 1923982, DE 2914330 (Madaus).

Also known dry extract of the fruits of the milk Thistle (Extr. cardui mariae fruct. siccum), which is derived from medicinal plants, using, inter alia, extracting agent ethyl acetate and lead in compliance with the standards Ph.Eur (Pharmacopoeia Calibration Standards).

In according to the specified requirements dry extract must contain preferably between 30 and 65% of the mass of silymarin (other ranges of content), silymarin contains the following fractions:

40-65% by weight: silybin(Jn) ( mixture, WITH 25 N 22 10 HE , with a molecular weight (MM) 482,4 g/mol)

10-20% by weight: ( mixture WITH 25 N 22 HE 10 , MM 482,4 g/mol)

20-45% by weight: and silidianin silikristin (WITH 25 N 22 HE 10 , MM 482,4 g/mol)

To extract the source material (in this case, a medicinal plant) usually degreased, subjected to extraction, filtered, concentrate and purify.

At the specified continuous extraction with the use of ethyl acetate/ethanol/acetone/methanol (optional in aqueous solution or water mixture with the solvents, usually spend filtering with the subsequent isolation. Next, perform a cleanup with ethanol and hexane (further degreasing)will get a silymarin the above composition.

Such a composition provides release from 30 to approximately 40% silymarin (measured in accordance with Ph. Eur. 5.7; 2.9.3 (01/2006:20903 revised), for example, using the method of rotating baskets» (basket method) or apparatus with the paddle mixers).

However, there is a great need in increasing the degree of redundant silymarin extract from the original.

It is known that these have low solubility or even insoluble in water (solubility net of silymarin is approximately 0.08 mg/ml at pH 6.9). Due to such values, the degree of solubility of the release of these compounds, and there are actually bioavailability/absorption in the body of humans and mammals, is not sufficient.

To increase the share of release of attempts were made to obtain derivatives , for example, using polyatomic alcohols, amino sugars or esters or receiving complexes connections inclusion, such as a cyclodextrin (EP 0422497 B1 (Madaus)or complexing connections-for example, phosphatidylcholine.

However, the disadvantages of these methods is optional education physiologically foreign substances, causing side effects.

The prior art is also known that the degree of redundant silymarin can be increased with the use of substances-media, such as 1-vinyl-2-pyrrolidone, mannitol and other (EP 0722918 B1, US 5906991 (Madaus)). In addition, the use of wetting agents, such as (). The EP 1021198 B1 (Madaus) describes a method of co-precipitation of silymarin with PEG. However, the drawbacks of all the above methods is shortness of dosing, as well as the possibility of formation of foreign substances, which can lead to unexpected side effects.

The prior art is also known document JP 2003135023 And that describes the process of extraction of the seeds of the milk Thistle (Silybum spotted) by using anhydrous ethanol with subsequent spray drying.

Based on the foregoing, the present invention is getting improved extract from the fruits of the milk Thistle, in particular, extract which has a much higher degree of release of silymarin, while preserving the natural properties. An additional objective is to obtain the extract without the use of additives, additives, chemicals media or wetting additives.

Problem is solved by the method of obtaining of extract from the fruits of the milk Thistle, which includes the following stages:

(a medicinal plant is subjected to extraction using solvent, having moderate polarity (e.g. ethyl acetate, ethanol, acetone, methanol, not necessarily containing aqueous fractions), preferably with 40-80°With, especially preferably with 50-70 Degrees;

(b) is separated, preferably filtered;

(c) concentrate, preferably in a vacuum at stirring, at a temperature of less than 60 Celsius degrees, preferably less than 40 C, and C') optional washed with hot water;

(d) mixed with ethanol, preferably 96% or more of pure ethanol or a similar solvent polarity, then mixed with hexane or similar solvent polarity and concentrate, preferably at a pressure of 1 ... 100 mbar (of 7.5-750 mm Hg), the resulting phase of ethanol-water is removed;

(e) dry and need not crushed;

(f) process anhydrous alcohol, preferably ethanol;

(g) optional filtered and concentrated; and

(h) dry and optional crushed.

Suddenly it was found that the implementation of additional stages (f) leads to a significant increase in the degree of liberation of silymarin to 80% (see sample). This is a major advantage, because with this according to the present invention is achieved by reduction of dosage extract of milk Thistle. Another advantage is that the opportunity to achieve the same quality of the received product, which in previous studies it was not only in the use of additives, additives, chemicals media and wetting agents.

The term «anhydrous alcohol» on stage (f) preferably includes C1-C4 alcohols, in particular, preferably ethanol, 99% or even 99,5%.

Based on the foregoing, the invention also relates to a method for the extract of the fruit milk Thistle, has a degree of liberation of silymarin is equal to 80% or above, according to which the extract containing silymarin in the amount of 15-85%, by mass, in particular, 30-65% by mass, process anhydrous alcohol, optionally filtered and concentrated, then dried and optional crushed.

In the framework of the present invention, «silymarin» means a mixture of substances containing (at least) four substances: silybin, , and in different concentrations. The content of these substances in relation to each other, as well as the presence of a mixture of additives is not essential. However, it is preferable that these substances meet the requirements of Ph.Eur or DAB in the amended version that observed according to the present invention.

«The degree of release of silymarin 80% or higher» means that the active substances of at least 80% soluble in aqueous solution (standard Ph.Eur., see examples).

This advantage allows to achieve improved .

The obtained extract of milk Thistle is particularly suitable for the purposes of the present invention, as a result of stage (f) significantly reduces the number of crystalline factions formed in the extract, the extract of the fruit milk Thistle has essentially amorphous structure.

Thus, the invention refers to a new extract of the fruits of the milk Thistle or mixture having essentially amorphous structure (see the comparative tests of x-ray analysis, presented on Fig.1).

In one of the most preferred variant of realization of the invention belongs to a new extract of the fruits of the milk Thistle or composition, containing having amorphous structure, in which the content of the crystalline fraction is less than 20%, preferably less than 10%, it is most preferable to less than 7 percent, even 5%.

Next invention relates to medicine made from the extract of the fruits of the milk Thistle, corresponding to the invention, or its use for the treatment and prevention of disorders of the liver and gallbladder, in particular toxic lesions of the liver (alcohol, fatty liver), , for example, in case of poisoning mushrooms, acute liver failure, necrosis of the liver dystrophy liver cirrhosis, liver fibrosis, hepatomegaly, fatty liver, functional liver failure, hepatitis, in particular hepatitis C.

Also the invention relates to the pharmaceutical composition containing medicinal substance according to the present invention, consisting of the extract of the fruit milk Thistle according to the present invention.

Extracts of milk Thistle according to the present invention may be obtained in the form of pharmaceutical preparations in dosage forms. This means that the drug may exist in the form of individual servings, for example, tablets, pills, capsules, pills, suppositories and ampoules, content of active substances which may be one or multiple single doses. Dosage form may contain, for example, 1, 2, 3, or 4 single doses or 1/2, 1/3 or 1/4 of a single dose. A single dose, preferably contains the number of active substances, which corresponds to a single intake of the medicine and which typically corresponds to a day dose or half, a third or a quarter of the daily dose.

Under non-toxic, inert, pharmaceutically acceptable substances native understand solid, semi-solid or liquid diluents, excipients and adjuvants to obtain all types of compositions.

Preferred pharmaceutical forms include tablets, pills, capsules, pills, granules, suppositories, solutions, syrups, suspensions and emulsions.

Tablets, pills, capsules, pills, granules can contain an active substance or a substance with traditional chemicals carriers, such as a) excipients and diluents, such as starch, lactose, saccharose, glucose, mannitol and silicic acid; (b) binder, for example, carboxymethylcellulose, alginates, gelatin and polivinilpirrolidon; humidifiers, for example, glycerin; (d) raising agents, for example, an agar-agar, calcium carbonate and carbonate sodium; (e) retardants dissolution, for example, waxes, f) accelerators absorption, such as Quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol, glycerol monostearate; (h) adsorbents, such as kaolin and bentonite; and i) lubrication, for example, talc, stearaty calcium and magnesium, solid polyethylene glycols; or mixture of substances) (i).

Tablets, pills, capsules, pills, granules can be covered by the standard shells and coatings, not necessarily containing opalescent components, and can have such a composition, in which the active substance or substances arrive only in the intestines or, preferably in a local region of the intestine, not necessarily with the provision of prolonged action, which polymers and polymeric wax, for example, can be used as encapsulating compounds.

Active substance or substances may also exist in form, optional with one or more of the foregoing vehicles.

In addition to the active substance or substances candles may contain traditional water-soluble or water-insoluble particulates matter-media, for example, polyethylene glycols, fats, for example, cocoa butter and higher esters (for example, ether C14 alcohol and C16 fatty acid) or mixtures of these substances.

In addition to the active substance or substances, solutions and emulsions can contain traditional substances-media, such as solvents, soljubilizatory, emulgatory, for example, water, ethanol, isopropanol, , ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-, dimethylformamide, oil, especially cotton-seed oil, peanut oil, corn oil, olive oil, castor oil and sesame oil, glycerin, formaldehyde, , polyethylene glycols and esters of fatty acids and sorbitol, or mixtures of these substances.

Together with the active substance or substances suspension may contain traditional substances-media, such as liquid diluents, for example, water, ethanol and propylene glycol, agents, for example, ethoxylated , and esters sorbitol, cellulose, aluminium, bentonite, agar-agar and gum, or mixtures of these substances. These dosage forms can also contain dyes, preservatives and perfumes and flavorings, such as oil, peppermint oil and eucalyptus oil, and sweeteners, such as saccharin.

EXAMPLES AND A SHORT DESCRIPTION OF

The following examples are provided for illustration of the present invention.

Example 1

Comparative tests on the release of silymarin:

Comparative extracts (silibinin Ch.-B.: 194051, Ch.- B.: 7085i) were prepared in accordance with this description, the silibinin Ch.-B.: 7085i received by the method with extra stage (f).

Data on the release of the active substance (silymarin) at pH 7.5 and conditions specified in Ph.Eur. (Dissolution test solids, Ph.Eur 5.7; 2.9.3 (01/2006:20903)) are presented below:

Sample

Time sampling

number in %

Silibinin Ch.-B.: 194051

After 30 min

4,16

Silibinin Ch.-B.: 7085i

After 30 min

49,13

The results show that the treatment of anhydrous ethanol on stage (f) leads to the fact that previously poorly soluble mixture silibinina, consisting of amorphous and crystalline structures, goes into amorphous state (see Figure 1) (i.e. changes in the lattice), leading to an increase in solubility and release of the active substance.

This stage allows to receive the above extract, which has a value of release of the active substance equal to 80% from the total number of silymarin is calculated for silibinina, (HPLC - Ph.Eur. 01/2007:2071), after 30 minutes, because that way improves the solubility not only silibinina, but also other isomers of silymarin.

Figure 1 shows the comparison of x-ray analysis data, for silibinina Ch.-B.: 194051 and silibinina Ch.-B.: 7085i, reflecting the change of crystal structure (conditions correspond example 2).

Radiographic analysis was carried out on the diffractometer X'pert Pro MPD from PANalytical B.V. with a focus on the method of Bragg-Brentano and using a detector of X Celerator.

Further comparative tests are described below.

Example 2

Methodology

a) sample Preparation

Prepared two powdered sample products:

no 7233i obtained by the method according to the present invention, includes the stage (f); and

no 7232i obtained without the use stage (f)

a) Conducted analysis by the method of powder

Introduction portions of powder substances carried in glass capillaries with a diameter of 0.5 Lindemann mm

b) the Equipment and the experimental conditions

Diffractometer PANalytical X'pert PRO MPD with θ/θ , having a radius of 240 mm, with a parallel lens with a hybrid monochromator and transfer geometry with holders for capillaries and a mixer.

Radiation: Cu-K (k=1,5406 & A).

Power supply: 45-40mA.

Entrance slit with fixing beam when the 0.19 mm

Crack Soller 0,02 radian for the incident beam and the diffracted beam.

c) Detector X Celerator having a working length 2122.

Range 2: from 3 up to 60 c step 0.017 and time of measurement 1500 seconds per step.

d) Task

Obtaining x-ray structural chart using the method of powder. Determination of the degree of crystallization.

e) Methodology

The degree of crystallization is a massive share of the crystalline phase in a mixture of crystalline and amorphous phases defined by the index of crystallization Ci:

Ci=100[Xc(Xa+Xc)], where XC is a massive share of the amorphous phase.

Value XC determined by summing the squares of all the narrow peaks (in the case of the crystalline phase) in the range of angular range according to the study. Values Ha received by the determination of the area of the broad peaks or «» (amorphous phase).

Results

Figure 2 and 3 presents the graphs obtained in all of the specified measuring range. Used a sample of mixtures of crystalline and amorphous phases.

Totals Ci amounted to 7% for the sample №7233i (see Figure 2) and 24% for the sample №7232i (see figure 3).

1. Method of obtaining of extract the fruits of the milk Thistle, wherein the extract containing 15-85%, by mass, in particular 30-65%, by mass, of silymarin, process anhydrous C1-C4 alcohol, optionally filtered and concentrated, then dried and optional ground, where the degree of liberation of silymarin is 80% or higher.

2. Method of receipt of the extract of the fruit milk Thistle, wherein a medicinal plant is subjected to extraction by solvent, having moderate polarity (e.g. ethyl acetate, ethanol, acetone, methanol, not necessarily containing aqueous fractions), preferably with 40-80°, most preferably with 50-70 Degrees; b) is separated, preferably filtered c) concentrate, preferably in a vacuum with stirring, at a temperature of less than 60 Celsius degrees, preferably less than 40 C and C') optional washed with hot water; (d) mixed with ethanol preferably 96% or more of pure ethanol or a similar solvent polarity, then mixed with hexane or similar solvent polarity and concentrate, preferably at a pressure of 1 ... 100 mbar formed phase of ethanol-water is removed; e) dry and need not crushed; f) process C1-C4 anhydrous alcohol, preferably ethanol; g) optional filtered and concentrated; and (h) dry and need not crush, and degree of liberation of silymarin is 80% or above.

3. Method of receipt of the extract of the fruit milk Thistle, which has a share of release of silymarin 80 percent or higher, according to claim 2, characterized in that at the stage of (a) the solvent with moderate polarity choose from the group consisting of ethyl acetate, ethanol, and methanol, preferably ethyl acetate.

4. Fruit extract of milk Thistle for treatment and prevention of disorders of the liver and gall-bladder, where the extract receive according to the mode on any one of claims 1 to 3.

5. Fruit extract of milk Thistle according to claim 4, wherein the specified fruit extract of milk Thistle, essentially, has an amorphous structure.

6. Fruit extract of milk Thistle according to claim 4, wherein the specified fruit extract of milk Thistle, essentially, has an amorphous structure, the content of the crystalline fraction is less than 20%, less than 10%, preferably less than 7%.