RU2712079C1 - Liposomal drug for treating local radiation damages of skin - Google Patents

Abstract

FIELD: pharmaceutics.SUBSTANCE: invention refers to pharmaceutics, particularly to agents for treating local radiation damages of skin. Medicinal agent according to the invention in the form of liposomes includes an aqueous solution of recombinant human alpha-fetoprotein (rchAFP) with its content in amount of not less than 1.0 mg/ml, solution of phospholipids Lipoid S80 in propylene glycol, preserving agents – methyl paraben and propylparaben, and butylhydroxytoluene, in quantities specified in the patent claim.EFFECT: according to the invention, the liposomal drug is stable during storage and is characterized by high aggregation resistance both during manufacture and storage.1 cl, 3 dwg, 5 tbl

Description

The invention relates to pharmaceuticals, in particular, to agents for treating local radiation lesions of the skin with liposomes containing human alpha-fetoprotein, and can be used in pharmaceuticals and medicine.

Known liposomal drug for the treatment of local radiation lesions of the skin, presented in p. RF No. 2642957 by class. A61K 31/00, 2039/5555, c. 03/11/2016., Op. 01/29/2018, and selected as a prototype.

The known product includes an aqueous solution of human alpha-fetaprotein (rhAFP) with a content of 0.1 mg / ml to 1.0 mg / ml, a solution of phospholipids Lipoid S80 in propylene glycol and preservatives (parabens, methyl paraben and propyl paraben, amount not indicated). Additionally, the tool may contain from 0.004 mg / ml to 0.06 mg / ml granulocyte colony stimulating factor person. The resulting composition is an emulsion

A disadvantage of the known agent is that it does not have aggregation stability (stability). Meanwhile, aggregation stability is one of the key characteristics of any emulsion preparations, including liposomal ones.

Aggregation resistance is an urgent problem in the preparation of liposome dispersions. Even if it is supposed to store the drug in the form of a lyophilisate, it is necessary that during the time that the liposome dispersion awaits lyophilization in the production cycle, liposome aggregation is minimal

The objective is to ensure the aggregation stability (stability) of the drug.

The problem is solved in that in a liposome drug for the treatment of local radiation injuries of the skin, which includes an aqueous solution of recombinant human alpha-fetoprotein (rhAFP) with a content of at least 1.0 mg / ml, a solution of phospholipids Lipoid S80 in propylene glycol and preservatives - methyl paraben and propyl paraben, ACCORDING TO THE INVENTION, butyl hydroxytoluene is additionally introduced in the following ratio of components:

Recombinant Alpha-Fetoprotein  (102 ± 1) mg Phospholipid LipoidS80  (1 ± 0.05) g Propylene glycol  (10.0 ± 1.0) g Methyl parahydroxybenzoate (methyl paraben)  (200 ± 5) mg Propyl parahydroxybenzoate (propyl paraben)  (75 ± 5) mg Butylhydroxytoluene  (5 ± 0.5) mg Purified water up to 100 g

The introduction of a stabilizer such as butyl hydroxytoluene, which is an antioxidant preservative, into the liposomal drug for the treatment of local radiation injuries of the skin, inhibits the oxidation processes, therefore, prolongs their shelf life and shelf life, in the above amount, it can significantly increase the aggregation stability of the liposomal composition in the manufacture and in further ensure the stability of the liposome composition during storage.

The technical result is an increase in the aggregation resistance of a liposome drug.

The inventive liposomal drug has a novelty in comparison with the prototype, differing from it by such essential features as the introduction of butylhydroxytoluene as the aggregation stability stabilizer and the above quantitative ratio of the main components, which together provide the desired result.

The applicant does not know the technical solutions possessing the indicated distinguishing features, which together ensure the achievement of the desired result, therefore, he believes that the claimed liposome drug for the treatment of local radiation injuries of the skin meets the criterion "and" from the inventive step ".

The inventive liposomal drug for the treatment of local radiation injuries of the skin can be widely used in medicine and therefore meets the criterion of "industrial applicability".

The invention is illustrated in tables and pictures, which presents the following test results of the claimed funds for aggregation stability and test results of its therapeutic effectiveness:

tab. 1 - the results of studies of the stability of the obtained samples;

tab. 2 - aggregation stability of the obtained liposomes;

tab. 3 - assessment of the effect of a liposomal drug on the course of skin radiation burn in mice CD1 with local irradiation at a dose of 60 Gy according to clinical indicators, (M ± m) day;

tab. 4 - assessment of the effect of a liposomal drug on the skin condition in the area of the burn wound in mice 7, 21, and 35 days after local skin irradiation at a dose of 60 Gy with histological parameters (M ± m);

- tab. 5 is an assessment of the effect of a liposomal drug on mouse stem cells;

- FIG. 1 - depilated area of the skin of the back in the mouse before irradiation

- FIG. 2 - phased fixation of the animal and the formation of skin folds on the back to obtain a local radiation burn;

- FIG. 3 is a view of a radiation burn wound at various stages of mouse pathogenesis.

The inventive liposomal drug contains an aqueous solution of recombinant human alpha-fetoprotein (rhAFP), a solution of Lipoid S80 phospholipids in propylene glycol, preservatives - methylparaben and propylparaben, and butylhydroxytoluene in the following ratio of components:

Recombinant Alpha-Fetoprotein  (102 ± 1) mg Phospholipid LipoidS80  (1 ± 0.05) g Propylene glycol  (10.0 ± 1.0) g Methyl parahydroxybenzoate (methyl paraben)  (200 ± 5) mg Propyl parahydroxybenzoate (propyl paraben)  (75 ± 5) mg Butylhydroxytoluene  (5 ± 0.5) mg Purified water up to 100 g

To assess the effect of butylhydroxytoluene on the stability and aggregation stability of the liposomal drug, the applicant conducted experiments.

In the experiments, a comparative assessment was made of the stability and aggregation stability of the initial liposome composition and the liposome composition containing butylhydroxytoluene.

As mentioned above, stability is one of the key characteristics of any emulsion preparations, including liposomal ones.

The stability of the proposed liposome compositions was determined according to GOST 29188.3-91 "Cosmetic products. Methods for determining the stability of an emulsion ”The method is based on separation of the emulsion by centrifugation. Equipment used

Laboratory centrifuge Eppendorf and a set of test tubes; Water bath.

General purpose laboratory balance BIOSAN of the 2nd accuracy class with the largest weighing limit of 200 g according to GOST 24104;

Liquid glass thermometer according to GOST 28498 and normative and technical documentation with an interval of measured temperatures from 0 to 100 ° C with a division price of 1 ° C;

Two tubes were filled into 2/3 of the volume with the studied liposome compositions and weighed, the result was recorded to the second decimal place. The mass difference of the tubes with the emulsion should not exceed 0.2 g. The tubes were placed in a water bath and incubated for 20 min at a temperature of 22-25 ° C. Then the tubes were removed, wiped them dry from the outside and centrifuged for 5 min at a speed of 100 s ^-1 .

Then determined the stability of the compositions. If no clear delamination was observed, the contents of the tube were carefully poured onto a piece of thick white paper and the presence or absence of delamination of the composition was noted.

A liposomal composition was considered stable if, after centrifugation in tubes, no more than a drop of the aqueous phase or an oil phase layer of no more than 0.5 cm was observed, and also there was no delamination of the composition on a white paper sheet.

Aggregation resistance is an urgent problem in the preparation of liposome dispersions. Even if it is supposed to store the drug in the form of a lyophilisate, it is necessary that during the time that the liposome dispersion awaits lyophilization in the production cycle, the aggregation of liposomes is minimal. Therefore, for the hydration of lipids upon receipt of liposomes, various stabilizers are introduced into the aqueous phase that increase aggregation stability, such as urea (in a concentration from 0.1% to 5%), sucrose (up to 3%), maltose (up to 5%), and others. stability was evaluated by the relative change in the optical density of the liposome dispersion sample (8) at a wavelength of 650 nm before (D1) and after (D2) centrifugation at 11000g for 10 minutes:

Obviously, the smaller the relative change in the optical density of the sample, the more stable the estimated dispersion.

The discussion of the results.

Data on the stability of the obtained samples are shown below in table 1.

Thus, the use of butylhydroxytoluene improves the stability of the liposomal composition.

The results of determining the aggregation stability of the obtained liposome preparations are presented in table 2.

Thus, in comparison with the prototype, the use of butyl hydroxytoluene in the inventive drug allows to increase the aggregation stability of the liposomal composition by 34% during manufacture and to further ensure the stability of the liposomal composition during storage.

At the same time, the effectiveness of the action of a liposomal drug with butylhydrotoxitoluene introduced into it for the treatment of local radiation lesions of the skin was maintained, which was confirmed by experiments.

The purpose of these experiments was to study the specific pharmacological activity of a liposomal drug based on phospholipids, including recombinant human alpha-fetoprotein (rhAFP), during the course of radiation burn of the skin of the ShA degree according to clinical indicators and indicators of the state of stem cells of hair follicles of the skin.

A liposomal drug is (per 100 g of product):

Component Name Concentration Purified water (FS.2.2.0020.15) Up to 100 g Propylene glycol (TU 6-09-2434-81) (10.0 ± 1.0) g Phospholipid Lipoid S80 (TU 9154-004-77972323-200-2005) (1.0 ± 0.05) g Methyl Parahydroxybenzoate (200 ± 5) mg Propyl parahydroxybenzoate (75 ± 5) mg Butylhydroxytoluene (New Component) (5 ± 0.5) mg Recombinant Human Alpha-Fetoprotein (102 ± 1) mg

(LLC NPP Enzymatic Technologies)

Studies performed on mice. The experiment used healthy animals that had not previously been subjected to other experimental procedures. Groups were formed by the continuous sampling method. The studies were performed on male CD1 runoff mice (age 3 months at the beginning of the experiment, weight 24-28 g). The conditions of detention and the experimental procedure corresponded to the general rules of work using experimental animals [10].

Animals of each species were kept under conditions of a conventional type of vivarium under artificial lighting. The animals received high-grade granular food (BioPro LLC Alliance and PZK 90 OJSC Bogdanovichsky feed mill) and drinking water ad libitum. The basic rules for maintenance and care were in accordance with the “Sanitary and Epidemiological Requirements for the Design, Equipment and Maintenance of Experimental Biological Clinics (Vivariums)”, approved by the Decree of the Chief Sanitary Doctor of the Russian Federation of 08.29.2014, as well as the rules adopted by the European Convention for the Protection of Vertebrate Animals (Strasbourg, 1986).

For each animal species, two groups were formed: a control group, where animals were injected with water for injection to the site of radiation damage to the skin, and an experimental group, where the liposome drug was applied to the site of skin radiation damage. Groups of mice consisted of 35 animals each. Of these, 20 animals were used to record the clinical picture of a radiation burn, 15 animals for histological studies (5 animals on the 7th, 21st, 35th day after irradiation).

Getting a skin burn in mice.

The optimal model for the experimental assessment of radiation damage to the skin is a IIIA degree burn, since this causes damage to the epidermis and the skin itself, but the epithelial elements of the skin are preserved, which are the starting material for self-healing of the wound, and, at the same time, the clinical manifestations of the burn look bright enough. It was previously found that to obtain a IIIA degree burn, the skin of mice must be irradiated at a dose of 60 Gy. The procedure for obtaining a local radiation burn in mice and rabbits was carried out in accordance with known methods.

The day before irradiation in animals, depilation was performed on a skin site in the center of the back with an area of 9 cm in mice (Fig. 1), which shows the depilated area of the back skin in mice before irradiation.

Before irradiation, the mice were fixed on the tablet in the back-up position, a skin fold was formed on the depilated portion of the back using a special clamp (Fig. 2), then local targeted skin irradiation was performed in the fold area. In FIG. 2 shows the fixation of the animal and the formation of skin folds on the back to obtain a local radiation burn

Skin irradiation in mice was carried out using a gamma-ray device of the State Chelyabinsk Regional Clinical Oncological Dispensary Theratron® Equinox. This installation uses a gamma radiation source containing cobalt-60. For local skin irradiation in animals, additional collimators with corresponding holes were used. The dose rate in the area of the skin fold in mice was 1.87 Gy / min. The area of the formed burn in mice was 2 ± 0.2 cm ² .

The scheme for applying a liposomal drug to the site of radiation damage to the skin is as follows.

Transdermal application of a liposomal drug and water for injection at the site of radiation exposure was performed 5 minutes after irradiation, then daily for 5 days and then 7, 9, 11, 13 days after irradiation (10 applications in total) using a dispenser. In mice, the drug was applied in a volume of 0.04 ml per day (in total for 10 applications - 0.4 ml per animal).

The animals of the control group were injected with water for injection in place of the radiation exposure according to the same scheme and in the same volumes as the animals of the experimental group.

Observations of the clinical picture of the course of skin radiation burn

In each group, animals were assigned an individual number: mice were labeled using standard methods.

A clinical examination of each animal was performed daily for 60 days after irradiation. The animal was examined, visual signs of local radiation damage to the skin were evaluated and recorded in the journal. Every day, photographs were taken of the radiation burn zone in animals with the goal of objectively documenting the clinical manifestations of radiation damage to the skin. Photographing was performed using a SonyA-37 camera (Japan) using a TamronAF 90 mmF / 2.8 macro lens and a YongnuoYN460 flash. The following settings were used: aperture f 9-11, ISO 100 sensitivity, shutter speed 1/160. Focusing was carried out manually on the camera display using electronic zoom. The shooting scale was 1: 3. To achieve uniform lighting, a diffuser was used. Cataloging, viewing and processing of photographs was carried out using the program AdobePhotoshop LightroomCC 2014.

For each animal, the duration of the destructive and reparative phases, as well as the individual periods of these phases, were calculated from visual observations and photographic materials. The destructive phase includes a latent period, a period of dry epidermitis, a period of wet epidermitis; the reparative phase includes the period of the scab, the period of epithelization of the burn wound.

The duration of the latent period was calculated from the day of exposure to the appearance of erythema minus 1 day. The time from the appearance of erythema to the day of formation of wet exudate is the duration of the period of dry epidermitis. The time from the day the moist exudate appears to the day the scab forms is the length of the period of wet epidermitis. The time from the first day of formation of the scab to the day of the last detachment of the scab is the length of the scab period. The time of complete disappearance of a skin defect is the end of the epithelization process. The time from the last crust falling away until the epithelium appears, similar in structure and pattern to the epithelium of the unaffected area of the skin - the length of the period of restoration of the epithelium.

The clinical picture of skin radiation burn in mice under local irradiation at a dose of 60 Gy is presented in FIG. 3, where a view of a radiation burn wound at various stages of pathogenesis in a mouse is presented.

Here A is the latent period; B - dry epidermitis (erythema, edema); B - wet epidermitis; G is a scab; D - the clinical picture of a radiation burn after falling off the last crust (scab); E - epithelization of a burn wound.

After a latent period (Fig. 3A), peeling and mild swelling appeared on the depilated area of the back skin — dry epidermitis (Fig. 3B), followed by an exudation period (Fig. 3B), characterized by the formation of separate foci of serous-hemorrhagic discharge, shrinking into thin brown crusts. Subsequently, the foci of exudation merged with each other, forming a continuous crust (scab) of light brown color (Fig. 3G). Subsequently, after the last scab fall, the epithelization stage began (Fig. 3E).

Histological examination of the skin

In experimental studies, the protective effect of a liposomal preparation containing recombinant human alpha-fetoprotein (rhAFP) on mouse epidermal stem cells was evaluated in a local radiation burn model of IIIA severity level.

Histological studies of the skin in mice were performed on the 7th (destructive phase of local radiation injuries of the skin), 21st (reparative phase of local radiation injuries of the skin) and 35th (period of epithelization of the reparative phase) days after irradiation of animals using routine methods. The selection of animals for each examination period was carried out by random sampling. In this case, the selection of material for histological studies was performed after euthanasia of animals (by the method of cervical vertebra dislocation). The material was fixed in a 10% buffered formalin solution (pH = 7.2) and poured into a special Histomyx medium. Serial plane-parallel sections 4-5 microns thick were made from each block-cassette. Serial histological sections were dewaxed, stained with hematoxylin-eosin according to standard routine methods.

On histological preparations, the thickness, the number of epidermal cell layers in the burn wound zone and the number of hair follicles were determined. Analysis of linear values and areas of the studied structures and counting of hair follicles was carried out using a hardware-software complex for analysis and processing of images in microscopy - “VideoTest - Morphology 5.0”.

The area of the visual field was measured automatically using a hardware-software complex for analyzing and processing images in microscopy, consisting of a Motic VA-400T trinocular microscope, a digital color image input system based on a ProgRes SZ digital camera, 1 / 1.8 '' high resolution (not less than 2080 × 1542) and the software "VideoTest - Morphology 5.0", and amounted to 1.2 mm ² . The study parameters (thickness, number of epidermal layers in the burn wound zone and the number of hair follicles) were calculated in 10 fields of view at a magnification of × 400.

The epidermal stem cells were determined using an immunohistochemical reaction using Dako serum CD34, 1/65 dilution. The amount of CD34 ⁺ and CD34 ^- cells was determined in the preparations. The calculation of the studied parameters was carried out in 10 fields of view with an increase of × 400, the area of the field of view was 0.34 mm ² .

Statistical analysis. For the studied parameters, the arithmetic mean value and its standard error (M ± m) were calculated. The statistical significance of differences in the experimental groups was determined using t-student test. Differences were considered statistically significant at p≤0.05.

The results of the study. The criteria for the effectiveness of the tested chemicals are a reduction in the duration of radiation exposure of the skin (for example, faster recovery), or a decrease in the severity of damage (for example, the formation of a milder degree of burn), a longer latent period, the absence of periods of wet epidermitis, scab in groups of irradiated animals receiving the test substance with respect to the irradiated untreated animal.

The results of a study of the effect of a liposomal drug on the course of a radiation burn of the skin.

The results of clinical observations made using visual and photo-observation of a radiation burn of the skin of the IIIA degree in stock CD1 mice are presented in table 3.

The complete healing of the burn wound from the moment of irradiation to the restoration of the epithelium in the irradiated control group was 45.4 ± 1.7 days. The duration of the latent period was 9.2 ± 0.4 days, dry epidermitis - 1.9 ± 0.2 days, the duration of wet epidermitis - 7.9 ± 0.3 days, the duration of the scab period - 6.4 ± 0.8 days . The duration of the destructive phase in animals of the control group was 19.2 ± 0.3 days. Starting from 26.5 ± 0.8 days, the period of epithelization of the burn wound (restoration of the epithelium) was observed in animals, which lasted 18.9 ± 1.3 days. The duration of the reparative phase in animals of this group was 26.2 ± 1.8 days.

When applying a liposome drug to the skin in the area of radiation exposure, there was a statistically significant shortening of the scab period by 50% with respect to the indicator in the irradiated control (t = 3.45; p = 0.0014). The duration of the destructive phase in animals of this group statistically significantly decreased relative to the control by 6% (t = 2.07; p = 0.046), the duration of the reparative phase decreased by 14%, but this difference did not reach statistical significance. The total healing time of a radiation burn of the skin was 41.1 ± 1.4 days, which is 6% less than in the control, but the level of statistical significance at t _article = 1.9 was p = 0.065.

Thus, according to the indicators of the duration of the periods of the course of radiation burn of the skin of the IIIA degree in mice, the study showed that the use of a liposomal drug had a beneficial effect on the course of the radiation burn, which was manifested in a reduction in the duration of the destructive phase and a reduction in the scab period compared to the parameters in the irradiated control group , earlier onset of epithelization of a burn wound.

In histological studies, the thickness and number of rows of epidermal cells were determined, the number of hair follicles in the field of view. Table 2 presents the results of histological studies of the skin condition in mice.

In mice of the biological control group, the number of rows of cells in the epidermis was 2.07 ± 0.20, the thickness of the epidermis was 19.1 ± 1.8 μm, the number of hair follicles was 13.3 ± 0.7 in the field of view of 1.2 mm ² . Irradiation at a dose of 60 Gy led to a change in these indicators (group control radiation + water). On the 7th, 21st and 35th, an increase in the epidermal layers was recorded 1.4 times, respectively (t = 2.56; p = 0.03), 4.5 (t = 14.86; p <0, 0001) and 2 times (t = 5.17; p = 0.0009) in relation to biological control. The epidermal thickness was increased on the 7th, 21st and 35th days, respectively, 2 times (t = 3.58; p = 0.0071), 9 times (t = 16.75; p <0.0001) and 3 times (t = 6.01; p = 0.0003) with respect to the control. The number of hair follicles in the field of view (1.2 mm ² ), on the contrary, was reduced by 8 times on the 7th, 21st and 35th days (t = 16.07; p <0.0001), 3 times (t = 8.84; p <0.0001) and 9 times (t = 15.49; p = p <0.0001), respectively.

When transdermal application of a liposome drug on a burn wound 5 minutes after irradiation, within 5 days and on 7, 9, 11, 13 days after irradiation, on the 7th and 21st day of the examination, the indicators in the group “Irradiation + drug ”Did not differ from the indicators in the“ Irradiation + water ”group. However, on the 35th day after the irradiation, statistically significant differences in the histological indices in the animals treated were recorded from the irradiated untreated control (control group (irradiation + water)): an increase in thickness of 2.2 times was recorded, the number of epidermal layers was 1.9 times, which can be interpreted as an increase in the proliferative activity of skin cells at the site of radiation damage. The number of hair follicles in the field of view of mice treated with a liposome drug was 2.6 times higher than that of untreated mice (table 4).

The results of evaluating the effect of liposome drug on stem cells of the skin of mice are presented in table 5.

In mice in the biological control group, the number of CD34 + cells was 33.1 ± 4.3 in the field of view, the number of CD34- cells was 121 ± 8.0, i.e., in the epidermis, stem cells make up about a third of the total number - - 0.28 ± 0 , 03. Irradiation at a dose of 60 Gy led to a statistically significant (t = 5.43; p = 0.0006) decrease in the number of CD34 + cells to 60% of the parameter in the biological control on the 7th knock after irradiation, later their number was restored to the level control. The number of CD34 cells on the 7th and 35th day did not differ from the control, and on the 21st day after irradiation, a statistically significant (t = 5.43; p = 0.0006) increase of 2.3 times was recorded.

In the dermis, changes caused by irradiation in the stem cell population of hair follicles were more pronounced than in the epidermis. The number of CD34 + cells at all stages of the study was statistically significant (respectively, t = 10.38, p <0.0001; t = 10.33, p <0.0001; t = 10.45, p <0.0001) and accounted for 2-3% of the indicator of biological control. A 2-fold decrease in CD34 cells on the 7th day was also recorded (but did not reach statistical significance), on the 21st and 35th days, 5 (t = 3.29; p = 0.01) and 10 (t = 3.73; p = 0.01) times, respectively.

In general, in the skin (epidermis + dermis) after irradiation at a dose of 60 Gy, a significant decrease in CD34 + cells by 15 times (t = 10.71; p <0.0001) is recorded on the 7th day, 4 times (5 t = 8 , 11; p <0.0001) on 21 and 3 times (t = 7.52; p = 0.0001) on the 35th day after irradiation. The number of CD34 cells was statistically significantly reduced only on day 35 (t = 3.84; p = 0.005).

The transdermal use of a liposomal drug for the treatment of radiation damage to the skin in mice led to a statistically significant increase in CD34 + cells by 4 times in the epidermis on the 35th day, by 4 times in the dermis (in the hair follicles), 21st in 6.7 times and 35th by 14 times relative to indicators in mice with a local untreated burn. In general, in the skin (epidermis + dermis) in irradiated mice treated, with respect to untreated mice, a statistically significant increase in the content of CD34 + cells was observed at all stages of the study. At the same time, the increase in CD34 + cells is more pronounced not on the 7th day after skin irradiation, but in the longer term - on the 21st and 35th days. On the basis of what, it can be concluded that it is not so much the protective effect of the liposomal drug that is triggered, as stimulating the proliferation of stem cells. At the same time, a 1.5-fold increase in the amount of CD34-β in the skin (epidermis + dermis) is recorded.

In well-known works it is shown that both stem cells of the epidermis and stem cells of hair follicles participate in the repair of skin lesions. These cells possess the properties of clonogenicity, are multipotent and migrate into the epidermis both physiologically and during regeneration. Thus, treatment with a liposomal drug provides, when the skin is irradiated, greater preservation and restoration of hair follicles, including stem cells, which contributes to faster skin restoration and is a positive effect of the drug.

Thus, immunohistochemical studies showed that, in general, in the skin (epidermis + dermis) in irradiated mice treated with a liposomal drug, in relation to untreated mice, a statistically significant increase in the content of CD34 + cells was observed at all stages of the study. Moreover, an increase in CD34 + cells is expressed at all times after irradiation of the skin, and in a more distant time, an increase in the number of CD34- is more pronounced. In rabbits, a similar pattern is observed. Based on what, it can be concluded that not only the protective effect of the liposomal drug on skin stem cells is triggered, but also stimulates their proliferation, which leads to an increase in their production - CD34-cells.

Based on the results of the experiments, the following conclusions were made:

1. In experiments on mice, it was found that, according to clinical indicators, the claimed liposomal drug has a therapeutic effect on the course of radiation burn of the IIIA degree skin in mice with transdermal administration within 14 days after irradiation at the site of radiation damage to the skin, which is expressed in a reduction in the duration of the wet period epidermitis, the period of the scab, the onset of the epithelialization stage and, thus, can be used to treat skin lesions (radiation dermatitis) caused by the action of ionizing radiation

2. According to histological indicators, the use of the inventive liposomal drug for the treatment of radiation burn leads to an increase in the proliferative activity of epidermal cells (in terms of thickness and number of layers of epidermal cells), provides greater safety and restoration of hair follicles, as well as stem cells in the skin.

Thus, the inventive liposomal drug for the treatment of local radiation injuries of the skin with an aggregation stability stabilizer introduced in it in the form of butylhydroxytoluene is aggregatively stable in comparison with the prototype and retains its therapeutic effectiveness.

Claims (2)

Liposomal drug for the treatment of local radiation injuries of the skin, including an aqueous solution of recombinant human alpha-fetoprotein (rhAFP) with a content of at least 1.0 mg / ml, a solution of phospholipids Lipoid S80 in propylene glycol and preservatives - methyl paraben and propyl paraben, characterized in that butylhydroxytoluene is additionally introduced into it in the following ratio of components: Recombinant Alpha-Fetoprotein  (102 ± 1) mg Phospholipid LipoidS80  (1 ± 0.05) g Propylene glycol  (10.0 ± 1.0) g Methyl parahydroxybenzoate (methyl paraben)  (200 ± 5) mg Propyl parahydroxybenzoate (propyl paraben)  (75 ± 5) mg Butylhydroxytoluene  (5 ± 0.5) mg Purified water up to 100 g

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