RU2320360C1 - Method for preparing medicinal preparation comprising copper chlorophyll analogues - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method involves extraction of rod-shaped gum-tree leaves with 95-96% ethanol at temperature 18-24°C for 1-2 days without stirring for 1-2 days in the mass ratio of rod-shaped gum-tee to ethanol = 1:(5-6) followed by settling of prepared product at temperature 6-8°C, not above, for 2-3 days, its filtration and the following treatment of prepared extract with copper salt at temperature 18-24°C for 6-24 h. As copper salt method involves using cuprous dichloride dihydrate or cuprous dichloride dihydrate 5-10% alcoholic solution in the mole ratio chlorophyll in prepared extract to cuprous dichloride dihydrate = 1:(3-4). Invention provides preparing the more active and stable product.

EFFECT: improved preparing method of drug.

10 tbl, 6 dwg, 6 ex

Description

The claimed invention relates to methods for producing drugs by the method of alcohol extraction of natural raw materials, namely rod eucalyptus (Eucaliptus viminalis. L), belonging to the myrtle family (Myrtaceae), and subsequent processing of the alcohol extract with a divalent copper salt to obtain a native drug complex containing copper analogues chlorophyll.

The drug obtained according to the claimed method has the trade name "Galenophyllipt" (registered trademark according to the certificate of the Russian Federation No. 286561, priority date 06/30/2004) and is an alcohol solution for oral and topical use, intended for use in medicine as an effective antimicrobial agents, in particular in the presence of antibiotic-resistant strains of Staphylococcus aureus (Staphyllococcus aureus).

Known methods for producing various drugs and biologically active additives (BAA) containing copper analogues of chlorophyll, by extraction of various types of natural raw materials, such as needles (Russian patent No. 200440266, priority date 12/29/1992), aspen bark (Russian patent No. 20034555, date priority 13.02.1992), propolis - raw and multicolor daisy grass (Russian patent No. 2066191, priority date 06.06.1992), kelp algae (USSR copyright certificate No. 1782603, priority date 11.16.1990), mint leaves (USSR copyright certificate No. 288869 , date pr priorities 12/08/1968 Inventor's Certificate of the USSR №955929, priority date 24.09.1980).

There is also a known method for producing metal-containing derivatives of bacteriochlorophyll and a pharmaceutical composition based on them (Russian patent No. 2193038, priority date 11.24.1996).

There is also known a method for producing metal-containing derivatives of chlorophyll and pharmaceutical compositions based on it (US patent No. 5,744,598, date publ. 04/28/1998).

Of the several types of eucalyptus (myrtle family) cultivated in Russia, medicine uses rod Eucalyptus and Eucalyptus ball, used to obtain antimicrobial, antiseptic, anti-inflammatory and other drugs (MD Mashkovsky. "Medicines." - M., vol. 1, 2, 2002, p. 316, 399 "Medicinal plant material. Pharmacognosy." / Edited by G.P. Yakovlev and K.F. Blinova, St. Petersburg: SpecLit, 2004, p.149-152).

The main biologically active substances extracted from eucalyptus leaves by extraction with 95% ethanol and other methods are essential oil (the main components are cineole and pinenes), phenol aldehydes, chlorophylls, etc.

In medicine, as an anti-inflammatory and antiseptic, tincture of eucalyptus (Tinctura Eucalypti) is used, obtained at a ratio of raw materials and solvent (1: 5) on 70% ethanol (MD Mashkovsky. “Medicines.” - M., 2002, t .1, p. 316).

From the leaves and shoots of eucalyptus pterygoid, the preparation Eucalymin containing phenolaldehydes is obtained, which is used in medical practice as a drug with a bacteriostatic effect on staphylococci, streptococci, diphtheria bacillus. In acute suppurative inflammatory processes, a 1% alcohol solution of Eucalymin is used (M.D. Mashkovsky. “Medicines.” - M., 2002, v. 2, p. 399 and “Medicinal plant material. Pharmacognosy.” Under Edited by G.P. Yakovlev and K.F. Blinova. - St. Petersburg: SpecLit, 2004, p. 152).

Known drug under the trade name "Chlorophyllipt", which is obtained from the leaves of ball eucalyptus (Eucalyptus globulus L) or rod eucalyptus (Eucaliptus viminalis. L) by extraction with 95% ethanol (MD Mashkovsky. "Medicines". - M., 2002, v.2, p. 399 and USSR author's certificate No. 801341, priority date, 04/14/1979, column 1, paragraph 2, above), which has antibacterial activity. In medical practice, use 1% alcohol or 2% oily solutions of Chlorophyllipt. The patent literature describes methods of using “Chlorophyllipt,” a 1% alcohol solution in medical practice (USSR copyright certificate No. 240932, priority date 10/22/1965 and USSR copyright certificate No. 463706, priority date 05/18/1973).

The method of obtaining “Chlorophyllipt” (1% alcohol solution) from rod-shaped eucalyptus (Eucaliptus viminalis. L) is described in USSR author's certificate No. 801341, priority date 04/14/1979, as well as on the Internet at the Innovation Technology Center MSTU im. N.E.Bauman at http://www.mgtu-sistema.ru/patents/1111215275/3.php, which is selected as a prototype of the claimed invention.

1% alcohol solution of “Chlorophyllipt” is obtained according to USSR author's certificate No. 801341 by extraction of leaves of the eucalyptus rod (Eucaliptus viminalis. L) with 96% ethanol and subsequent processing of the obtained concentrated extract with a divalent copper salt, namely 4% copper sulfate solution, followed by extraction of the target product - the amount of hydrophobic components, including copper analogues of chlorophyll, benzene. Extraction with benzene is carried out three times with stirring each time for 15 minutes and settling for 5-6 hours. The settled benzene extract is washed 3 times with purified water and evaporated under vacuum at a temperature of 40 ° C. According to the description of the USSR author's certificate No. 801341 (column 2, paragraph 1 below), “Chlorophyllipt” (1% alcohol solution), obtained from rod eucalyptus by the same technology, is 2 times more active bactericidal drug compared to “ Chlorophyllipt ”with a 1% alcohol solution isolated from ball eucalyptus.

As can be seen from the description of the production technology of “Chlorophyllipt” (1% alcohol solution) according to USSR author's certificate No. 801341 during the preparation of the preparation, the intermediate product of the extraction of rod-shaped eucalyptus containing chlorophyll is subjected to both heat treatment and chemical treatment with a poisonous substance - benzene, as well as mechanical processing (mixing), which leads to a decrease in the quality of the target product due to the loss of its native properties, as well as a decrease in the stability of the drug due to thermal destruction of the extracted chlorophyll and its copper analogues and a decrease in the antibacterial activity of the drug, especially in relation to Staphylococcus aureus.

The fact of the destruction of chlorophyll as a result of thermal exposure is known from the prior art, namely from RF patent No. 2034555 (page 3, paragraph 2 below). The fact of the low activity of "Chlorophyllipt" (alcohol solution 1%) in relation to Staphylococcus aureus is also known from the prior art, for example, from RF patent No. 2066191 (page 1, paragraph 3 above).

In addition, as a result of studies of a 1% alcohol solution of the drug “Chlorophyllipt” obtained by the method according to the copyright certificate No. 801341, carried out by the authors of the present invention, it was experimentally established that when processing the intermediate product extract, the consumption of copper sulfate CuSO ₄ · H ₂ O in 4 times exceeds its actual need for the chemical reaction of copper plating of chlorophyll.

Thus, the authors of the present invention found that in the prototype in the reaction mixture, copper sulfate is in excess, and therefore it can under certain conditions, for example at high temperatures, react not only with chlorophyll, but also with other components of the alcoholic leaf extract rod-shaped eucalyptus, for example phenolic aldehydes, components of essential oil and others, which leads to a decrease in the native (natural) properties of the target product.

The objective of the present invention is to provide a new method of obtaining a medicinal product from the leaves of eucalyptus rod, in which the resulting medicinal product would retain all the main biologically active components, that is, had a high degree of nativeness, was stable even during prolonged storage, and was obtained by technology that excludes the use of toxic solvents as well as thermal, chemical and mechanical effects that can lead to changes and destruction of the active components of the drug That and a decrease in its activity.

The problem is solved in that the drug is obtained by extraction of leaves of eucalyptus rod-like ethanol and subsequent processing of the extract with a salt of divalent copper. New is that the extraction of the leaves of eucalyptus rod is carried out with 95-96% ethanol for 1-2 days without stirring at a temperature of (18-24) ° C with a mass ratio of leaves of eucalyptus rod-shaped to ethanol 1: (5-6) with sedimentation of the obtained product for 2-3 days at a temperature not exceeding (6-8) ° C, filtering it and subsequent processing of the extract with a salt of divalent copper for 6-24 hours at a temperature of (18-24) ° C, while as a salt copper use copper dichloride 2-water or 5-10% alcohol solution of copper dloris that 2-aqueous with a molar ratio of chlorophyll contained in the obtained extract to copper dichloride 2-aqueous 1: (3-4).

The whole set of new features in the claimed invention is essential to achieve the task.

Carrying out the technological process of obtaining the drug in the absence of heating of the extract of leaves of eucalyptus rod-like (intermediate product), the absence of mechanical influences when mixing the reagents during the extraction and washing, as well as in the process of carrying out the chemical reaction of copper-plating of chlorophylls, allows to maintain a high degree of nativeness of the obtained drug “Galenofillipta "And its high biological activity due to the exclusion of factors, significantly influenced chlorophylls decomposing.

The authors of the present invention conducted comparative studies of the preparations “Chlorophyllipt” (alcohol solution 1%) and “Galenofillipt”, during which the electronic absorption spectra of both drugs, the pH value, the effect of irradiation with daylight on the optical density of both drugs, as well as their qualitative and quantitative composition by chromatography in a thin layer of sorbent. In addition, the content of the main active substances: phenol aldehydes, cineole, pinene, copper analogues of chlorophyll was determined in the compared preparations.

As a result of experimental studies of the drug "Chlorophyllipt" (alcohol solution 1%), carried out by the authors of the present invention, it was found that, unlike "Galenophyllipt," there is no pinene, a valuable biologically active substance extracted with ethanol from the leaves of eucalyptus rod-like, and the content copper analogues of chlorophyll, cineole and phenol aldehydes are reduced. Comparative data on quality indicators (preserving the drug’s nativeness) of “Chlorophyllipt” (alcohol solution 1%) and “Galenophyllipt” are shown in table 1.

As can be seen from table 1, in the preparation "Galenofillipt" obtained in accordance with the claimed method, the content of the main biologically active substances: phenol aldehydes, cineols, copper analogues of chlorophyll is 2-3 times higher than in the preparation "Chlorophyllipt" (alcohol solution 1% ) obtained by the method selected as a prototype. In addition, the pH of Galenophyllipt is 1.4 times greater than the pH of Chlorophyllipt (1% alcohol solution). The latter can accelerate the decomposition of chlorophylls, because it is known that at low pH values the stability of chlorophylls decreases (Berezin B.D. Coordination compounds of porphyrins and phthalocyanine. - M .: Nauka, 1978).

Table 2 shows the results of a study of the effect of irradiation with daylight on the optical density of the alcohol extract from the leaves of the eucalyptus rod and Galenophyllipt and Chlorophyllipt (1% alcohol solution). We studied two preparations “Chlorophyllipt” (alcohol solution 1%) (manufacturer Ukraine) and “Chlorophyllipt” (alcohol solution 1%) (manufacturer Russia), presented on the Russian market and obtained in the same way using the prototype technology. At the same time, the Chlorophyllipt drug (alcohol solution 1%) of the series No. 490705 manufactured in Ukraine and the Chlorophyllipt drug (alcohol solution 1%) of the series No. 71205 made in Russia were used for comparative studies.

Compared samples of the preparations were irradiated with daylight for 7 days. As can be seen from table 2, the decrease in optical density in the drug "Galenofillipt" after 7 days of exposure to daylight was 9%, for the drug "Chlorophyllipt" (alcohol solution 1%) (Ukraine) - 26%, for the drug "Chlorophyllipt" (alcohol solution 1%) (Russia) - 35%. The data obtained indicate that the stability of the drug "Galenophyllipt" obtained by the claimed method is 2.8-3.8 times higher than both compared drugs obtained by the method of the prototype.

It is known from the prior art that upon the interaction of divalent copper salts with individual chlorophyll in alcohol solutions, a hyperchromic and hypochromic shift of the first absorption band of chlorophyll from (662 ± 3) nm to (650 ± 3) nm is observed in the electronic absorption spectrum of the mixture, which is typical for copper chlorophyll analogues (Journal of the Uspekhi Khimii RAS Magazine No. 73 (2) 2004, article by BD Berezin et al. “Chemical transformations of chlorophyll and its use to create environmentally friendly dyes of a new generation”, p. 202, Tables 4, 3 line above).

In the process of implementing the present invention, it was of scientific and practical interest to establish how this interaction proceeds in multicomponent alcohol solutions containing, along with chlorophyll, also other chemicals capable of various reactions with divalent copper. Such systems include alcoholic extract of the leaves of eucalyptus rod, containing, in addition to chlorophylls, phenolaldehydes, essential oil and other components.

When adding increasing amounts of an alcoholic solution of copper dichloride 2-water with a known concentration to the alcoholic extract of eucalyptus leaves with a known chlorophyll content, we found in the mixture (Fig. 1) a gradual decrease and then the disappearance of the first absorption band of chlorophyll at (662 ± 3 ) nm and a simultaneous gradual increase in the absorption maximum at (650 ± 3) nm (copper analogues of chlorophyll).

Unexpectedly, the authors of the present invention found that this process ends when the molar ratio of chlorophyll and copper dichloride 2-water 1: (3-4), respectively. Further addition of copper dichloride 2-water (1: 4 and 1: 5) does not change the form of the electronic absorption spectrum (figure 1).

For all ratios of chlorophyll and copper dichloride 2-water, there was no change in the electronic absorption spectrum of the mixture in the range 220-290 nm, characteristic of phenol-aldehydes (figure 2). These data allow us to conclude that in the technological process for the preparation of the drug Galenophyllipt according to the claimed invention, the interaction of copper dichloride 2-water only with chlorophyll occurs, and other components of the extract of eucalyptus leaves, such as phenolaldehydes, components of essential oil (cineole and pinene ) do not react with copper dichloride 2-water. The unexpectedly established phenomenon described above allows one to obtain a medicinal product according to a new technology while preserving all the beneficial properties of the main biologically active components of the eucalyptus leaves.

It is known from the prior art that the purification of chlorophyll derivatives in the manufacture of drugs is a very labor-intensive technological process requiring a large consumption of solvents and high energy consumption (RF patent No. 2147203, p. 1, paragraph 4, above). The optimal selection of technological parameters of the proposed method for producing the drug "Galenophyllipt" eliminates the stage of purification of the extract with an extremely dangerous solvent - benzene, and also eliminates the stage of removing excess copper salt by washing with water, benzene residues - using repeated heating, extraction and azeotropic distillation of benzene with alcohol, which allows you to maintain high biologically active properties of the resulting drug by preserving the native properties of its components s and to ensure its stability.

The essence of the invention is illustrated by the following drawings:

figure 1 - electronic absorption spectra of an alcoholic extract of leaves of eucalyptus rod-like and products of copper plating at various molar ratios of chlorophyll and copper dichloride 2-aqueous;

figure 2 - electronic absorption spectra of an alcoholic extract of leaves of eucalyptus rod-like and copper-plating products at various molar ratios of chlorophyll to copper dichloride 2-aqueous in the region of (200-280) nm;

figure 3 - diagram of the chromatograms of alcoholic extract of eucalyptus rod (E), "Galenofillipta" (G), "Chlorophyllipt" (alcohol solution 1%) (X) on the plates Sorbfil PTSX-P-A-UV in the solvent system acetone-petroleum ether (3: 7);

figure 4 - electronic absorption spectra of chlorophylls contained in alcoholic extracts from the spots of the chromatogram of leaf extract of eucalyptus rod-shaped;

figure 5 - electronic absorption spectra of chlorophylls contained in alcoholic extracts from the spots of the chromatogram "Galenophyllipt";

6 - electronic absorption spectra of chlorophylls contained in alcoholic extracts from the spots of the chromatogram "Chlorophyllipt" (alcohol solution 1%).

Here are examples of specific implementation of the proposed method of the invention.

Example 1. The technology for the preparation of the drug "Galenophyllipt", a solution for oral and topical use of alcohol, with a molar ratio of chlorophyll in the extract-intermediate and copper dichloride 2-aqueous 1: 3, respectively.

To prepare 1000 l of the drug "Galenophyllipt" according to the claimed method, 200 kg of crushed leaves of eucalyptus rod (FSP 42-0039-3375-02) are taken, loaded into the extractor and filled with 95% ethanol (FS 42-3072-00) with a mass ratio of eucalyptus leaves rod-like to ethanol 1: 5 in the amount of 1175 liters, taking into account the coefficient of alcohol absorption of raw materials to obtain 1000 l of the intermediate product (extract-intermediate), and produce extraction at room temperature (18-24 ° C) for 24 hours without stirring. The resulting intermediate extract must be stored in the dark.

After 24 hours, the obtained extract-intermediate was poured into a reactor cooled by brine and defended for 2 days at a temperature of + 8 ° C. Then, a sample is taken from the thus obtained extract-intermediate, and the content of chlorophyll, phenol aldehydes, solids, alcohol, pH value is determined, the electronic absorption spectrum is taken (Fig. 1, curve 1 “extract”). Upon receipt of the analysis data that meet the requirements for the extract-intermediate, the latter is filtered through a corrugated cardboard on a filter press. In the electronic spectrum of the filtered extract-intermediate there is only one absorption band with a maximum (662 ± 3) nm, characteristic of chlorophyll (Fig. 1 curve 1 “extract”).

The filtered extract-intermediate is copper-plated with the calculated amount of a 10% alcohol solution of copper dichloride 2-aqueous at a molar ratio of chlorophyll contained in the extract-intermediate to copper dichloride 2-aqueous 1: 3, respectively, which is calculated on the basis of the mass ratio of the extract-intermediate to copper dichloride 2-water is 1000: 0.076. Then the copper-plated extract-intermediate is maintained at a temperature of from 18 to 24 ° C for 24 hours, after which the electronic absorption spectrum of the obtained copper-coated product is removed in the region from 600 to 700 nm.

In the electronic absorption spectrum, an absorption band is observed with a maximum at (650 ± 3) nm (copper analogues of chlorophyll), as well as an increase in optical density to 0.36 (Fig. 1, curve 5 (1: 3)).

After 24 hours, a sample of the obtained target product “Galenophyllipt” is taken and analyzed in accordance with the requirements of the pharmacopeia article in the sections: description, authenticity, pH, heavy metals, alcohol content, dry residue, microbiological purity, quantitative determination of phenol aldehydes, copper content, toxicity.

Upon receipt of satisfactory results of the analysis, the obtained "Galenofillipt" is filtered through a corrugated cardboard on a filter press. Then "Galenofillipt" is transferred to packaging in bottles of dark glass and packaging in a pack of cardboard.

As a result of the implementation of the proposed method receive the drug "Galenophyllipt", a solution for oral administration and local use of alcohol of the following composition:

Eucalyptus twig leaves - 200 g Copper Dichloride 2-Aqueous - 0.06 g Ethyl alcohol 95% to receiving 1000 ml of the drug

Example 2. A method of obtaining a medicinal product containing copper analogues of chlorophyll, with a molar ratio of chlorophyll in the extract-intermediate to copper dichloride 2-aqueous as 1: 0.5.

200 kg of eucalyptus leaves are extracted by 95% ethanol as described in Example 1 in this application, prior to the stage of copper-plating of the intermediate product extract with copper 2-hydrous chloride. The filtered extract-intermediate is copper-plated with 2-hydrous copper at a ratio of chlorophyll contained in the resulting extract-intermediate to 2-aqueous copper dichloride as 1: 0.5, which, in terms of the mass ratio of the extract-intermediate and copper dichloride 2-aqueous, is 1000 : 0.0126. Then the copper-plated extract-intermediate is maintained at a temperature of from 18 to 24 ° C for 24 hours.

Then remove the electronic absorption spectrum in the region from 600 to 700 nm.

A wide absorption band is observed in the electronic absorption spectrum, including two weakly expressed absorption maxima: chlorophyll at (662 ± 3) nm and the copper analog of chlorophyll at (650 ± 3) nm, which indicates only partial binding of chlorophyll to copper dichloride 2-water due to insufficient the amount of the latter (Fig. 1, curve 2 (1: 0.5)).

Example 3. A method of obtaining a medicinal product containing copper derivatives of chlorophyll, with a molar ratio of chlorophyll in the extract-intermediate to copper dichloride 2-aqueous as 1: 1.

200 kg of eucalyptus leaves are extracted by 95% ethanol as described in Example 1 in this application, prior to the stage of copper-plating of the intermediate product extract with copper 2-hydrous chloride. The filtered extract-intermediate is copper-plated with 2-hydrous copper at a ratio of chlorophyll contained in the obtained extract-intermediate to 2-hydrous copper of 1: 1, which, in terms of the mass ratio of the extract-intermediate to copper 2-hydrous, is 1000: 0 , 0253. Then the copper-plated extract-intermediate is maintained at a temperature of from 18 to 24 ° C for 24 hours.

Then remove the electronic absorption spectrum in the region from 600 to 700 nm.

In the electronic absorption spectrum, a decrease in the intensity of the absorption band of chlorophyll in the region of (662 ± 3) nm, as well as an increase in the optical density of absorption at (650 ± 3) nm as compared with curve 2 (Fig. 1) 0.24 to 0.27, which indicates an increase in the yield of copper analogues of chlorophyll, formed with an increase in the added amount of copper dichloride 2-aqueous (figure 1, curve 3 (1: 1)).

Example 4. A method of obtaining a medicinal product containing copper analogues of chlorophyll, with a molar ratio of chlorophyll in the extract-intermediate to copper dichloride 2-aqueous as 1: 2.

200 kg of eucalyptus leaves are extracted by 95% ethanol as described in Example 1 in this application, prior to the stage of copper-plating of the intermediate product extract with copper 2-hydrous chloride. The filtered extract-intermediate is copper-plated with 2-hydrous copper at a ratio of chlorophyll contained in the obtained extract-intermediate to 2-hydrous copper of 1: 2, which in terms of the mass ratio of the extract-intermediate to copper 2-hydrous is 1000: 0.051 . Then the copper-plated extract-intermediate is maintained at a temperature of from 18 to 24 ° C for 24 hours.

After that, the electronic absorption spectrum is removed in the region from 600 to 700 nm.

In the electronic absorption spectrum, there is a sharp decrease in optical density at (662 ± 3) nm and an increase in absorbance at (650 ± 3) nm, from 0.27 to 0.35 (Fig. 1, curve 4 (1: 2)) , which indicates a significant increase in the content of copper analogues of chlorophylls.

Example 5. A method of obtaining a medicinal product containing copper analogues of chlorophyll, with a molar ratio of chlorophyll in the extract-intermediate to copper dichloride 2-aqueous as 1: 4.

200 kg of eucalyptus leaves are extracted by 95% ethanol as described in Example 1 in this application, prior to the stage of copper-plating of the intermediate product extract with copper 2-hydrous chloride. The filtered extract-intermediate is copper-plated with 2-aqueous copper chloride at a ratio of chlorophyll contained in the obtained intermediate product to 2-aqueous copper dichloride as 1: 4, which in terms of the mass ratio of the extract-intermediate product to copper 2-aqueous copper is 1000: 0.101 . Then the copper-plated extract-intermediate is maintained at a temperature of from 18 to 24 ° C for 24 hours.

Then remove the electronic absorption spectrum in the region from 600 to 700 nm.

In the electronic absorption spectrum, there is a maximum absorption at (650 ± 3) nm, its optical density does not change (0.36), which indicates the complete binding of all chlorophyll to copper 2-aqueous dichloride (Fig. 1, curve 6 (1: 4 )).

Example 6. A method of obtaining a medicinal product containing copper analogues of chlorophyll, with a molar ratio of chlorophyll in the extract-intermediate to copper dichloride 2-aqueous as 1: 5.

200 kg of eucalyptus leaves are extracted by 95% ethanol as described in Example 1 in this application, prior to the stage of copper-plating of the intermediate product extract with copper 2-hydrous chloride. The filtered extract-intermediate is copper-plated with 2-hydrous copper at a ratio of chlorophyll contained in the obtained extract-intermediate to 2-hydrous copper of 1: 5, which, in terms of the mass ratio of the extract-intermediate to copper 2-hydrous, is 1000: 0.127 . Then the copper-plated extract-intermediate is maintained at a temperature of from 18 to 24 ° C for 24 hours.

Then remove the electronic absorption spectrum in the region from 600 to 700 nm.

Compared with the previous example, no changes in the electronic absorption spectrum are observed. In the electronic absorption spectrum, there is a maximum absorption at (650 ± 3) nm, its optical density does not change 0.36 (Fig. 1, curve 7), that is, the further addition of copper dichloride 2-water does not increase the yield of copper chlorophyll analogs. From the data obtained (examples 1-6) it follows that for complete copper plating of chlorophylls contained in the extract from the leaves of eucalyptus rod-like, it is necessary to use 3-4 equivalents of copper dichloride 2-aqueous.

The authors of the present invention conducted a comparative study of the chromatograms of the alcoholic extract of the leaves of eucalyptus rod, "Galenophyllipt" obtained by the claimed method, as well as "Chlorophyllipt" (alcohol solution 1%) (prototype).

When comparing the properties of the extract of leaves of eucalyptus rod with Galenophyllipt obtained from it, as well as with Chlorophyllipt (1% alcohol solution) on the chromatogram in a thin layer of sorbent, chlorophyll spots are detected in the extract (visually and in UV light with a maximum radiation of 365 nm). On the chromatograms of Galenophyllipt and Chlorophyllipt (alcohol solution 1%), visually emerald spots of copper analogues of chlorophyll with various Rf values are found - the dimensionless ratio of the distance that the spot of the photographed substance passed from the start to the distance traveled by the solvent system from start to finish ( figure 3). Moreover, in the chromatograms, the positions of the chlorophyll spots of the extract and the spots of copper analogues of chlorophyll in Galenofillipt coincide and are in the upper parts of the chromatograms, which indicates that the structure of the molecule after the replacement of Mg by Cu is preserved and indirectly confirms that the obtained substance is a copper analog of chlorophyll . In the case of “Chlorophyllipt” (1% alcohol solution), the number of spots of copper chlorophyll analogs is greater than the number of spots on the Galenophyllipt chromatogram and they have significantly lower Rf values, which indicates that during the production of “Chlorophyllipt” under chemical influence excessive amounts of copper sulfate and low pH, as well as intensive stirring during triple extraction with benzene and heating, the structure of chlorophylls changes (partial degradation) with the formation of chromium atograficheski less motile derivatives, which indicates the receipt of drug with significantly lower native properties.

Further, the authors of the present invention obtained and studied electronic absorption spectra of alcoholic extracts from individual spots of chromatograms of the eucalyptus rod, Galenophyllipt and Chlorophyllipt extract (1% alcohol solution) (Figs. 4, 5, 6).

As can be seen from figure 4, in the electronic absorption spectrum of alcoholic extracts from the spots of the chromotogram of the eucalyptus leaf extract, absorption maxima at 662, 655 and 651 nm, characteristic of chlorophylls, are detected.

In the electronic absorption spectrum of alcoholic extracts from spots of Galenophyllipt chromatograms, spots of copper chlorophyll analogues with maxima of 650, 640 and 647 nm are found, and in the electronic absorption spectra of alcoholic extracts from spots of chromatograms of Chlorophyllipt (1% alcohol solution) with maxima at 650 (2 spots) and 640 nm.

The number of copper chlorophyll analogs present in Galenophyllipt, calculated from the area of the absorption bands of the spectra, is about 3 times greater than in Chlorophyllipt (1% alcohol solution). The data obtained indicate that the content of copper analogues of chlorophyll in "Galenophyllipt" is significantly higher than in "Chlorophyllipt" (alcohol solution 1%), which suggests a higher biological activity of the drug "Galenophyllipt" obtained according to the claimed method, in comparison with the drug "Chlorophyllipt" (alcohol solution 1%) (prototype).

We present the results of preclinical comparative studies of the antimicrobial activity of the preparations "Galenofillipta" and "Chlorophyllipta" (alcohol solution 1%).

The study of antibacterial and antifungal properties in experiments in vitro.

The studies were carried out in accordance with the “Methodological guidelines for the study of the antimicrobial activity of pharmacological substances”, “Methodological guidelines for the study of the antifungal activity of pharmacological substances” (in the book “Manual on the experimental (preclinical) study of new pharmacological substances.” - M .: Medicine, 2005, p. 515-531 and p. 582-588).

Quantitative assessment of the antimicrobial effect of the drug Galenophyllipt in comparison with the drug "Chlorophyllipt" (alcohol solution 1%) and known antibiotics.

To identify the benefits of the Galenophyllipt drug, a quantitative assessment of its antibacterial effect was carried out in comparison with Chlorophyllipt (alcohol solution 1%) and six antibiotics using the dilution method.

Used museum strains of microorganisms occupying a different systematic position: gram-positive cocci - Stafilococcus aureus-209 P; gram-negative sticks - Escherichia coli-88; spore-forming bacilli - Clostridium dificille-4938 and Candida albicans-674 fungi, as well as field strains of Pseudomonas aeruginosa and Salmonella typhi suis.

The results obtained, presented in table 3, indicate that the minimum inhibitory concentration of "Galenophyllipt" for all tested strains of microorganisms is from 0.2 to 2.0 μg / ml, i.e. comparable to the effect of known antibiotics. However, unlike them, "Galenofillipt" has a much wider spectrum of action, as it is able to suppress the vital activity of fungi (C.albicans). In the case of “Chlorophyllipt” alcohol solution of 1%, the minimum inhibitory concentration is much larger and ranges from 1.3 to 8.0 μg / ml, i.e. the activity of Chlorophyllipt (alcohol solution of 1%) is significantly lower than Galenophyllipt.

Therefore, the drug "Galenophyllipt" has a pronounced antibacterial effect in concentrations comparable to effective doses of antibiotics. It has a wider spectrum of action compared to most antibiotics used in medical practice.

Compared with the drug "Chlorophyllipt" (alcohol solution 1%) (prototype), the drug "Galenofillipt" is about 2 times more effective in antibacterial activity.

The study of the antibacterial action of the compared drugs by agar culture (GF XI, issue 2, p. 188 and “Change” No. 2 to it dated 01.01.2002)

Three series of studies were carried out by inoculation on agar (in Petri dishes with a diameter of 90 mm) and liquid culture media (in test tubes) in relation to the control strains: E. Coli (ATCC 25922), St.aureus (ATCC 6538-P), Ps. aemgmosa (ATCC 9027), Vas. Subtilis (ATCC 6633), Vas. cereus (ATCC 10702), and C. albicans mushrooms (ATCC 885-653). For this, dilutions of 1:10, 1:20, 1:50, 1: 100, 1: 200, 1: 500 were prepared from 10 ml of each preparation using a phosphate buffer solution. 1 ml of preparations and their dilution were added to cups and test tubes, where growth of test microorganisms was taken into account after incubation. Similar volumes of distilled water were added to the control.

The research results are presented in table 4.

As can be seen from table 4, in the control samples there was an active growth of test microorganisms.

The experimental results indicate a wide spectrum of antimicrobial activity in the drug "Galenofillipt", which turned out to be significantly more active than the comparison product "Chlorophyllipt" (alcohol solution 1%).

The study of the biocidal action of the compared drugs.

In the course of the study, the effect of the compared preparations "Galenofillipt" and "Chlorophyllipt" (alcohol solution 1%) on the culture of microorganisms was studied.

Test cultures were selected according to the principle of their different sensitivity to adverse environmental influences, including to the studied drugs, due to: the type of structure of the cell wall (gracile, pharmicut, yeast); the presence on the cell wall of additional surface layers (capsules, etc.); the origin of the strain (museum or clinical isolate); individual characteristics of the microorganism; the density of the population of microorganisms experiencing the "pressure" of an adverse factor.

As an additional indicator, the belonging of the test culture to normal (NM) or opportunistic (UPM) microflora was taken into account.

The list and characteristics of test cultures are shown in table 5.

Test cultures were grown in appropriate culture media (Lactobacillus acidophilus in MPC-1; Escherichia coli M-17, Klebsiella pneumonia, Salmonella enteritidis, Staphylococcus aureus 209 in nutrient broth; Candida albicans in maltose broth) for 24 hours at 37 ° FROM. Standard suspensions with a density of 103-108 CFU / ml were prepared from liquid cultures and 100 μl were applied to the surface of similar agar media containing 0.025-0.5% of the studied substances. Crops were incubated at 37 ° C for 5 days, scanning daily, after which colonies were counted. The results were expressed in lgKOE / ml (table 6).

In the absence of colony growth, it was believed that 100 μl of the sample contains less than 1 colony, i.e. less than 10 colonies in 1 ml. Accordingly, lgKOE / MA = l is correctly represented as lgKOE / ml <1.

As can be seen from table 6, the presented results indicate that Galenophyllipt has a wide spectrum of antimicrobial activity against yeast and bacteria, regardless of the type of structure of their cell wall and population density. The effective concentration of the Galenophyllipt drug correlates with the degree of resistance of microorganisms to adverse external influences (0.05% for moderately stable and 0.1% for resistant microorganisms). Gracillicut bacteria and yeast in the studied concentration range are resistant to Galenophyllipt, regardless of the density of the microbial population.

Therefore, the drug "Galenophyllipt" has a biocidal effect on representatives of both normal and conditionally pathogenic microflora, superior in this respect to the comparison drug "Chlorophyllipt" (alcohol solution 1%).

Investigation of the specific anti-burn and antimicrobial activity of the Galenophyllipt and Chlorophyllipt preparations (1% alcohol solution) in a model of localized purulent-inflammatory infection in in vivo experiments.

The study was carried out in accordance with the "Methodological recommendations for experimental (preclinical) study of drugs for the local treatment of purulent wounds and burns." - M.: Ministry of Health of the USSR, 1989, 48 p.

The studies were carried out in the form of comparing the effectiveness of the therapeutic effect of Galenophyllipt, Chlorophyllipt (1% alcohol solution) and Dermazin, an antibacterial cream with silver sulfadiazine (Lek Pharmaceutical and Chemical Company dd, Slovenia), which is widely used in practice for local treatment of burns (registration number 005398 dated 10/14/94 in the Register of Medicinal Products of Russia; ND: 42-298-94) on the nature and rate of post-burn regeneration. In an experiment on laboratory animals (white rats, males), burns of the same depth and area were simulated. The differences between the compared series were only in the applied therapeutic agents. Objectivity of the research results was achieved by identical conditions for all animals for their animals, treatment, as well as a unified methodology for evaluating the data obtained.

Modeling of thermal burns was carried out as follows.

Limited in area, skin burns were modeled in rats using light radiation using a special device developed by specialists of the Military Medical Academy named after S.M. Kirov and allowing accurate dosing of the area and degree (depth) of a burn lesion. Our research used its modified version. On the clipped skin of the back of each rat (symmetrically to the midline of the back) using the installation, two sections of the burn 2.5 × 2 cm each were created. The total area of symmetrically located burns was 10 cm ² , that is, about 4% of the body surface area. The depth (degree) of the burn is 3 "a" and "b" degrees.

The methods of using the compared drugs in all animals were the same - the application of Galenophyllipt and Chlorophyllipt (1% alcohol solution) in the form of tampons soaked with them, or Dermazin cream in a dose of 150 mg / 10 cm ² or 800-900 mg / kg was carried out on the burned surface in a day throughout the entire period of healing of the wound surface. Dry sterile wipes were applied to the control animals on the burn sites, which changed after 1-2 days, and all manipulations (including applying the burn) were performed under short ketamine anesthesia.

The following were recorded: indicators of the general condition of the animals (food and motor activity); indicators of the course of the wound process (burn area and the end of epithelization). Be sure to carry out histological monitoring of the healing process.

As can be seen from the results of the studies, the motor and nutritional activities of all animals did not change significantly, which can be explained by the small size of the burn, and therefore the low severity of the developing general changes. Therefore, the main object of study, as required by the objectives of the study, was the dynamics of local changes in burn wounds.

Evaluation of the results of treating burns, presented for clarity in the form of tables 7 and 8, indicates that, compared with untreated animals (control), Galenofillipt, Chlorophyllipt (alcohol solution 1%) and Dermazin had a clear positive effect on rate of regeneration.

A comparative assessment of the rate of rejection of a burn scab showed that, according to this indicator, the effectiveness of the compared products increased in the series: “without treatment” <“Chlorophyllipt” (alcohol solution 1%) <“Dermazin” <“Galenofillipt”. A similar trend was observed in other indicators of clinical efficacy.

It should be noted that in all respects, Galenophyllipt was significantly more effective than the control, and in terms of the rate of rejection of the scab and the rate of complete epithelization, it significantly exceeded both Dermazin and Chlorophyllipt (alcohol solution 1%). In addition, while with the use of Dermazin cream every second animal showed the formation of a keloid scar, then in animals treated with Galenophyllipt, the formation of keloids was not observed and the burns healed by first intention.

Table 7 shows the results of the complete healing of burn wounds at different periods of observation. By the 25th day, complete epithelization of burn wounds in rats treated with Galenophyllipt occurred - in 76%; “Dermazin” - in 55%; “Chlorophyllipt” (alcohol solution 1%) - in 50%; in untreated - in 18%. By the 30th day of treatment, these figures were respectively: 93%, 85%, 79% and 25%.

Consequently, the regularity noted above, i.e. the therapeutic effectiveness of the tested drugs in terms of complete healing of the burn increased in the series: "without treatment" <"Chlorophyllipt" <"Dermazin" <"Galenofillipt".

The results are confirmed by pathomorphological studies of burn wounds.

In the group of animals without treatment, a scar consisting of dense collagenic connective tissue fibers is visible at the burn site on days 35-37.

The morphological data are confirmed visually by the dynamics of the repair of burn wounds, which is presented in photographs of experimental animals.

According to a histological study, the use of the “Galenophyllipt” preparation in conditions of light burn of 3 “a” and “b” degrees contributes to faster and more complete epithelization of the burn surface, while in conditions of burn without the use of the drug, a keloid scar forms and recovery appendages of the skin does not occur.

In the process of using the compared preparations and the course of the burn process on day 5, 10 and 20, the general condition of the animals was evaluated, including 11 standard indicators: body weight, urine protein, urine specific gravity, blood sugar, serum protein, ALT, AcT activity, white blood cell count, erythrocyte count, hemoglobin content, ESR.

The results of the study indicate the presence of dynamic changes characteristic of the course of inflammatory and reparative processes: at the beginning of burn intoxication, the rate of weight gain is slowed down, the content of red blood cells, hemoglobin is reduced, leukocytosis and ESR are increased. In the future, these indicators gradually return to normal levels, and there is a tendency to faster normalization in the treatment of burns "Galenofilliptom."

The study of antimicrobial activity of the compared drugs.

The study of the bactericidal and bacteriostatic properties of the studied preparations was carried out by agar diffusion (disk method) and by introducing it into the agar well in relation to the control strains: E. Coli (ATCC 25922), St.aureus (ATCC 653 8-P), Ps. aemginosa (ATCC 9027), Vas. Subtilis (ATCC 6633), Vas. cereus (ATCC 10702), and C. albicans mushrooms (ATCC 885-653). In accordance with the Global Fund XI, issue 2, p. 187, 3 series of studies were conducted.

The results are presented in table 9.

As can be seen from table 9, in the control samples (with physiological saline) around the disks there was a continuous growth of test microorganisms.

When using the compared drugs, growth retardation of microorganisms was observed, which indicates the sensitivity of microorganisms to them.

The sterility of the studied preparations was also controlled by direct inoculation on Saburo liquid medium (State Pharmacopia XI, issue 2, p. 188). At the same time, prolonged incubation did not lead to clouding of the nutrient medium, which indicates the sterility and bactericidal properties of the compared preparations.

In addition, seeding of wound burn surfaces was sown on blood agar, Chistovich, Saburo, Endo and meat and peptone agar. In this case, growth of staphylococci and gram-negative rods was revealed (table 10), which disappeared in the case of “Chlorophyllipt” (alcohol solution 1%) only by the 20th day, and in the case of “Galenofillipt” - by the 10th day.

As can be seen from the data in experiments with a non-extensive burn (3 "a" and "b" degrees) on an area of about 4% of the body surface, without having a noticeable effect on the general condition, the Galenofillipt preparation contributed to the optimization of the reparative process to a greater extent. than "Dermazin" and "Chlorophyllipt" (alcohol solution 1%) used in practical medicine as a means of local treatment.

Acceleration of the start and complete rejection of the scab, the beginning and end of epithelialization compared with the course of the same processes when using “Dermazin” and “Chlorophyllipt” (alcohol solution 1%), allows us to note the great effectiveness of the drug “Galenofillipt” in achieving normal, close to optimal flow of reparative regeneration processes.

"Galenofillipt" can be used as an agent in the treatment of burns of 3 "a" and "b" degrees, point or circular deep burns or linear deep burns with an area of not more than 4% of the body surface. "Galenofillipt" is not inferior, and in many indicators of the clinical course of the burn wound, it surpasses the prototype drug "Chlorophyllipt" and ointment "Dermazin".

Thus, as can be seen from the above data, it has been reliably established that the drug "Galenophyllipt" obtained according to the claimed method has a higher biological activity, higher nativeness and higher stability of the dosage form compared with the drug "Chlorophyllipt" (alcohol solution 1 %) obtained by the prototype method.

Table 1 Comparison of quality indicators (nativeness) of “Chlorophyllipt” and “Galenofillipta” Quality indicators (native) of the drug Galenophyllipt Chlorophyllipt Galenophyllipt / Chlorophyll Comparison Results
lipt Copper derivatives of chlorophyll (D = 650 nm) ~ 0,045 ~ 0.012 > 2-3 times Phenolaldehydes,% ~ 0.82 ~ 0.46 > 2 times Cineol,% 0.12 0,03 > 2-3 times Pinen,% 0,03 - yes / no pH 4.3 3.1-3.2 > 1.4 times

table 2 The effect of irradiation with daylight on the optical density of the drug "Galenofillipt" and the drug
“Chlorophyllipt” (alcohol solution 1%, production Ukraine) and “Chlorophyllipt” (alcohol solution 1%, production Russia). Drug name Original optical density Optical density after 7 days of exposure Residual density Density drop Galenophyllipt at 650 nm 0.391 at 650 nm 0.356 91% 9% Chlorophyllipt, alcohol solution 1% (Ukraine) at 650 nm 0.233 at 650 nm 0.173 74% 26% Chlorophyllipt, alcohol solution 1% (Russia) at 650 nm 0.139 at 650 nm 0.090 65% 35%

Table 3 The minimum inhibitory concentration (MIC) for the compared in vitro preparations (μg / ml) A drug Test microbe E.coli Ps. St. Cl. C. S.typhis Galenophyllipt 2.0 2.0 0.2 0.2 2.0 2.0 "Chlorophyllip" (alcohol solution 1%) 8.0. 4.0 1.5 1.3 4.8 5.0 Benzylpenicillin 15.0-60.0 * stable 0.003-60.0 0.018-6.0 stable 1.2-12.0 Erythromycin 10.0-100.0 * stable 0.1-100.0 * 0.1-5.0 stable 43.0 Oleandomycin stable stable 0.3-100.0 * 1.0-20.0 stable stable Streptomycin 0.5-100.0 0.5-200.0 * 0.5-200.0 1.0-200.0 stable 0.004-20.0 Tetracycline 0.5-100.0 * 3.0-100.0 * 0.1-100.0 * 0.1-100.0 stable 1.0-2.5 Chloramphenicol 0.5-200.0 * 10.0-200.0 * 0.5-100.0 * 1.0-50.0 stable 0.8-5.0 * - and more; the second interval of minimal inhibitory concentrations of antibiotics is given for resistant strains of pathogens.

Table 4 The results of inoculation of test microorganisms when using the preparations "Galenofillipt" and "Chlorophyllipt" (alcohol solution 1%) Microor test
ganisms Dilution of the drug The control 1:10 1:20 1:50 1: 100 1: 200 1: 500 ". Galageno phillipt Chloro phillipt Galageno phillipt Chloro phillipt Galageno phillipt Chloro phillipt Galageno phillipt Chloro phillipt Galageno phillipt Chloro phillipt Galageno phillipt Chloro phillipt E.Coli +++ - - - - - - - ± - + --- ++ St.aureus +++ - - - - - - - + - ++ --- ++ Ps.aeruginosa +++ - - - - - ± - + - ++ --- ++ You.Subtilis +++ - - - - - - - + - ++ --- ++ You. Cereus +++ - - - - - - - + - ++ + + Mushrooms C. albicans +++ - - - - - ± - + - ++ ++ +++ Designations: +++ - continuous growth; ++ - growth within 100 colonies; + - growth of single colonies; ± - weak or slow growth; - - lack of growth.

Table 5 Characterization of the used test cultures Test culture Type of cell wall structure Representative of NM or UPM Museum strain or clinical isolate Resistance to adverse environmental influences Escherichia coli Gracilicut NM Museum moderate M-17 Klebsiella Gracilicut UPM Clinical high pneumonia isolate (capsules) Salmonella Gracilicut UPM Museum weak enteritidis Staphylococcus Firmikutny UPM Museum moderate aureus 209 Lactobacillus Firmikutny NM Museum moderate acidophilus D75 Lactobacillus Firmikutny NM Museum moderate acidophilus D76 Candida Yeast UPM Clinical high albicans isolate

Table 6 The survival of microorganisms in the presence of compared drugs, depending on the concentration of drugs and density
microbial population Test culture The control Galenophyllipt,% Chlorophyllipt,% 0.02 0.05 0.1 0.25 0.5 0.0 0.05 0.1 0.25 0.5 Escherichia coli M-17 8 8 one one one one 8 3 9 one one 7 6 one one one one 6 2 one one one 6 5 one one one one 5 2 one one one 5 four one one one one four 2 one one one four 2 one one one one 2 one one one one 3 one one one one one one one one one one Klebsiella pneumonia 8 8 8 3 one one 8 8 5 3 2 7 7 7 2 one one 7 7 four 2 one 6 6 6 one one one 6 6 four 2 one 5 5 four one one one 5 four 3 2 one four four 3 one one one four 3 3 2 one 3 3 2 one one one 3 3 2 one one Salmonella enteritidis 8 7 one one one one 8 four 3 2 2 6 5 one one one one 5 3 2 2 one four 3 one one one one 3 2 2 one one Staphylococcus aureus 209 8 8 one one one one 8 5 3 2 2 7 7 one one one one 7 5 2 2 one 6 6 one one one one 6 2 one one one 5 5 one one one one 5 2 2 2 one four four one one one one four 2 2 one one 3 3 one one one one 3 2 2 one one Lactobacillus acidophilus D-75; 7 one one one one one 3 2 one one one 6 one one one one one 3 2 one one one 5 one one one one one 2 2 one one one four one one one one one 2 one one one one 3 one one one one one 2 one one one one Lactobacillus acidophilus D-76 8 8 one one one one 8 5 3 2 2 7 7 one one one one 7 3 2 one one 6 6 one one one one 6 9 2 one one 5 5 one one one one 5 2 2 one one four four one one one one four 9 one one one 3 3 one one one one 3 9 one one one Candida albicans 8 8 7 one one one 8 7 5 3 2 7 7 6 one one one 7 6 four 2 2 6 6 5 one one one 6 5 3 9 2 5 5 four one one one 5 four 2 2 one four four 3 one one one four 3 2 one one 3 3 one one one one 3 2 one one one

Table 7 Clinical criteria for the effectiveness of the compared drugs in a burn wound - the healing time of burn wounds in rats, days (M ± m) Experimental group Elimination of perifocal reaction Wound cleansing (scab rejection) The appearance of granulation Beginning of marginal epithelization End of healing (complete epithelization) The control 4.5 ± 1.0 12.5 ± 1.0 17.2 ± 1.4 17.0 ± 1.9 46.4 ± 3.6 Dermazin 2.2 ± 0.4 7.9 ± 0.6 * 13.1 ± 2.0 13.2 ± 1.1 31.5 ± 1.9 * "Chlorophyllipt" (solution 2.5 ± 0.3 8.1 ± 0.4 * 12.1 ± 1.6 14.4 ± 1.3 33.5 ± 1.7 * alcohol 1%) Galenophyllipt 1.4 ± 0.4 * 5.2 ± 1.3 * 6.9 ± 1.1 * 8.3 ± 0.9 * 22.0 ± 1.5 * * - significant differences from control (p <0.05)

Table 8 Percentage of fully epithelized burn wounds when using comparable drugs Experimental 25th day of observation 30th day of observation Group Burn without treatment - dry sterile dressing eighteen% 25% Dermazin 55% 85% "Chlorophyllipt" (alcohol solution 1%) fifty% 79% Galenophyllipt 76% 93%

Table 9 Zones of growth inhibition of microorganisms (mm) when introduced into the culture discs with the studied drugs Microorganism "Chlorophyllipt" (alcohol solution 1%) Galenophyllipt Exceeding the delay zone,% E.Coli 18 ± 2 22 ± 3 22 St.aureus 15 ± 1 17 ± 4 13 Ps.aeruginosa 17 ± 4 23 ± 3 35 You.Subtilis 19 ± 2 25 ± 5 31 You. Cereus 13 ± 2 23 ± 3 77 Mushrooms C. albicans 9 ± 2 15 ± 4 67

Table 10 The results of crops detachable burn surface Experimental series Day after the burn 2nd 5th 10th The 20th Without treatment +++ ++ ++ ++ Dermazin ++ ++ - - "Chlorophyllipt" (alcohol solution 1%) +++ +++ + - Galenophyllipt +++ ++ - - where the notation is used: +++ - continuous growth; ++ - growth within 100 colonies; + - growth of single colonies; - - lack of growth.

Claims (1)

A method of obtaining a medicinal product containing copper analogues of chlorophyll by extraction of leaves of eucalyptus with ethanol and subsequent processing of the extract with a copper salt, characterized in that the extraction of leaves of eucalyptus with rod is carried out with 95-96% ethanol for 1-2 days without stirring at a temperature (18-24) ° C with a mass ratio of leaves of eucalyptus rod-shaped to ethanol 1: (5-6) with sedimentation of the obtained product for 2-3 days at a temperature not higher than (6-8) ° C, its filtration and subsequent processing the obtained extract with a copper salt for 6-24 hours at a temperature of (18-24) ° С, while copper dichloride 2-aqueous or 5-10% alcohol solution of copper dichloride 2-aqueous is used as a copper salt in a molar ratio chlorophyll contained in the obtained extract, to copper dichloride 2-aqueous 1: (3-4).

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