RU2465899C1 - Medication for treating purulent wounds - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to pharmaceutical industry, in particular to medication for treatment of purulent wounds by method of photodynamic therapy. Medication for treating purulent wounds by method of photodynamic therapy in form of gel, which includes zinc polycholinyl phthalocyanine, hydroxyethylcellulose, dimethylsulfoxide or benzalconium chloride (alkylbenzyldimethylammonium chloride), block-copolymer of ethylene oxide and propylene oxide or polyethyleneglycol, water, taken in specified quantity. Medication for treatment of purulent wounds by method of photodynamic therapy, representing film, obtained by drying of gel described above.

EFFECT: medication is convenient in application and ensures healing of purulent wounds in minimal possible terms.

2 cl, 4 tbl, 11 ex

Description

The invention relates to pharmaceutical chemistry, namely to means for antimicrobial photodynamic therapy (AFDT).

The treatment of local bacterial infections is one of the complex and far from solved medical problems. As a result, in many cases, treatment of local infections and purulent wounds requires a long time and is ineffective. The use of anti-inflammatory and antibacterial drugs, including antibiotics, for the treatment of diseases of microbial etiology leads to the emergence of chemotherapy-resistant microorganisms at a rate faster than the introduction of new effective drugs into practice. In this regard, AFDT is of great importance in the treatment of local infectious processes. The advantage of the AFDT method using phthalocyanine dyes is the absence of the development of resistance of microorganisms to this type of treatment. The combined effect of the photosensitizer and activating radiation, accompanied by the formation of singlet oxygen, leads to selective oxidative destruction of pathogenic microorganisms through multiple oxidation processes of various subcellular structures. The photodynamic damage caused by the cytotoxic effect of singlet oxygen leads to the death of viruses, fungi and bacteria, including antibiotic-resistant strains of Staphylococcus aureus, Pseudomonas aeruginosa and others. The antimicrobial effect of APDT does not decrease with time with long-term treatment of chronic local infectious processes, while the bactericidal effect has a local character and does not have a systemic destructive effect on the saprophytic flora of the body.

Effective AFDT sensitizers are known - phthalocyanines with cationic substituents, which have an affinity for a wide range of bacteria and are also used for the treatment of purulent wounds (RF patent No. 2282647, 47B 47/32, 08/27/2006, prototype). One of these photosensitizers is zinc polycholinylphthalocyanine with a degree of substitution of 6-8, a water-soluble dye having a maximum long-wavelength absorption in water at 680 nm. Describes the use of its solutions applied to damaged areas using a solution of gauze soaked in a solution. This method in many cases makes it difficult to accurately dose the drug. The introduced dose depends on the humidity of the napkin, on its tightness to all the treated areas and the loss of solution during its spreading. It should be borne in mind that a certain part of the drug is adsorbed on a napkin and does not enter the affected area.

The objective of the invention is to develop tools for the treatment of local infections of purulent wounds, convenient to use, ensuring the penetration of the photosensitizer into the treated tissue in a calculated dose at an economical consumption of the drug.

The solution to this problem is achieved by obtaining a gel or film based on it for the treatment of local infections of purulent wounds by the method of AFDT, including zinc polycholinylphthalocyanine, water-soluble polymers and excipients allowed for use in medicine in the following proportions of components, wt.%:

zinc polycholinylphthalocyanine 0.02-1.00

hydroxyethyl cellulose 1.3-1.6

dimethyl sulfoxide or benzalkonium chloride

(alkylbenzyldimethylammonium chloride) 0.2-3.0

block copolymer of ethylene oxide and propylene oxide or polyethylene glycol 3.2-4.0

propylene glycol 4.0-7.7

water is the rest.

Such ratios of the ingredients of the composition provide the greatest therapeutic effect of the resulting gels and films.

The tool was obtained by adding zinc polycholinylphthalocyanine synthesized according to the method described in the prototype and auxiliary components into a gel base prepared in advance by dissolving hydroxyethyl cellulose in water. In the case of obtaining a film, a gel containing zinc polycholinylphthalocyanine was applied evenly on a flat surface and dried.

The efficiency of the photosensitizer from gels and films to the treated tissues was determined by measuring their exogenous fluorescence using a contact method. The data obtained showed that the developed compositions quickly release zinc polycholinylphthalocyanine, which enters the wound tissue. Moreover, in experiments on mice, it was shown that the photosensitizer is mainly localized in the wound and surrounding tissues, enters the bloodstream within 2 hours after the start of application of the gel. In distant tissues, trace amounts of zinc polycholinylphthalocyanine are determined after 2, 4, and 6 hours of gel exposure. In the liver, with an increase in the application time, the normalized fluorescence (Ф _n ) of zinc polycholinylphthalocyanine increases, and in the kidneys it is recorded in trace amounts at all exposure times. In the spleen F _{n of the} drug is registered only after a six-hour application of the gel.

Using the claimed funds was conducted AFDT for a long time not healing purulent wounds.

The technique for the treatment of purulent wounds consisted of applying a gel to the wound surface for a period of 30 minutes to 3 hours. Then, the affected tissues were irradiated with a semiconductor laser with an exposure of 2 to 10 minutes. The use of laser light sources (with a wavelength of 650-670 nm) for the excitation of the photodynamic reaction and inactivation of purulent flora is the most effective. This is expressed in the rapid cleansing of wounds from the decay products and vital functions of microorganisms, the earlier appearance of healthy granulations and the relief of inflammatory phenomena in the wound. Of no less importance is the cosmetic aspect - the absence of gross cicatricial deformities with large soft tissue defects.

The procedure was carried out daily or every other day until the wound was completely cleansed of purulent discharge and wound detritus, from 3 to 6 procedures per treatment course, depending on the severity and prevalence of the purulent process.

Complete purification of the purulent wound occurred, as a rule, after 5-7 days with daily use of AFDT, while under normal conditions and the use of standard antibacterial and local treatment, these periods are on average from 2 to 3 weeks, and sometimes more.

The invention is illustrated by the following examples, but is not limited to.

Example 1

To 87.6 g of water (or 87.3 g in case of using a brand of 250 MR hydroxyethyl cellulose), 3.2 g of a block copolymer of ethylene oxide and propylene oxide (emuxol-268) are added with stirring, the mixture is stirred at 60 ° C, then cooled to room temperature, 1.3 g (Natrosol 250 type HHR) or 1.6 g (type 250 MR) are added to the hydroxyethyl cellulose and stirred for 2 hours, 7.7 g of propylene glycol, 0.1 g of benzalkonium chloride are added and mixed again . 0.02 g of zinc polycholinylphthalocyanine with a degree of substitution of 7.5 was added to the obtained gel base, and the mixture was stirred for 15 minutes. The compositions of the obtained gels are given in table 1.

Examples 2 and 3

The gels were obtained according to the method of example 1, but zinc polycholinyl phthalocyanine was taken in amounts of 0.2 and 1 g with a corresponding decrease in the amount of water. The compositions of the obtained gels are given in table 1.

Example 4

1.3 g of hydroxyethyl cellulose (Natrosol 250 type MR) was added to 87.7 g of water with vigorous stirring and the mixture was stirred for 30 minutes at 50 ° C. Then the mass is cooled to room temperature, 4.0 g of propylene glycol, 4.0 g of polyethylene glycol 400, 3.0 g of dimethyl sulfoxide are successively added to the resulting gel with stirring and stirred for another 15 minutes. Then, 0.02 g of zinc polycholinylphthalocyanine with a degree of substitution of 7.5 is introduced into the composition and the mass is mixed again. The composition of the obtained gel is given in table 1.

Examples 5 and 6

Films were prepared according to the method of Example 4, but zinc polycholinyl phthalocyanine was taken in amounts of 0.2 and 1 g with a corresponding decrease in the mass of water. The compositions of the obtained gels are given in table 1.

Table 1 The composition of the gels (wt.%) In examples 1-6 Example chol-pczn SCE (type) DMSO BH PEPO PEG PG Water one 0.02 1.3 (HHR) - 0.2 3.2 - 7.7 87.6 0.02 1.6 (MR) - 0.2 3.2 - 7.7 87.3 2 0.2 1.3 (HHR) - 0.2 3.2 - 7.7 87.4 3 one 1.3 (HHR) - 0.2 3.2 - 7.7 86.6 four 0.02 1.3 (MR) 3 - - four four 87.7 5 0.2 1.3 (MR) 3 - - four four 87.5 6 one 1.3 (MR) 3 - - four four 86.7 Notes
1. chol-PcZn - zinc polycholinylphthalocyanine, HEC - hydroxyethyl cellulose, DMSO - dimethyl sulfoxide, BH - benzalkonium chloride, PEPO - block copolymer of ethylene oxide and propylene oxide (emuxol-268), PEG - polyethylene glycol, PG - propylene.
2. Used SCE brand Natrosol 250.

Example 7

The gels of examples 4-6 are applied to a teflon surface with a layer of 2 mm thick and placed in an oven for 8 hours at 50 ° C. As a result, elastic transparent films of green color with a thickness of about 0.2 mm are obtained.

Example 8. Determination of the rate of release of polycholinylphthalocyanine zinc from the claimed films in the wound

The photosensitizer release was studied as follows. Pre-selected animals (BDF ₁ mice) a day before the study, wool from the thigh surface was removed using a depilatory cream. Before the start of the study, the animals were anesthetized (0.1 ml 0.25% droperidoli). To obtain a wound, a piece of skin was cut out on a mouse thigh with a diameter of 0.5 cm. An applicator film with zinc polycholinylphthalocyanine was applied on the wound surface and surrounding skin for 1-2 hours. After this time, the composition was removed from the wound surface of the animal. After 3-5 minutes, the mice were killed by dislocation of the cervical vertebrae, then tissue samples (wound surface, skin, muscle) were removed. Exogenous tissue fluorescence was measured ex vivo immediately after the killing of the animal. For each batch of the drug, material obtained from three animals was examined, and the data were compared with data obtained in a similar way for animals not exposed to the claimed films (control).

Fluorescence was recorded by the contact method on a LESA-06 laser electron-spectral setup. Fluorescence was excited by He-Ne laser radiation (wavelength 632.8 nm). The integrated intensity received by the fluorescence spectra in the range 645-736 nm was normalized by the integrated intensity of the diffuse scattering inverse signal of the exciting laser radiation fabric (λ _max = 632,8 nm), thus determining normalized fluorescence (F _m) of tissues.

From the data of table 2 it can be seen that the drug mainly accumulates in the wound, adjacent muscle and skin and is not found in the liver and distant muscles and skin. Increasing the time of application of the film of 1 to 2 hours leads to an increase in P _n photosensitizer in the wound, a similar pattern is observed in the surrounding skin.

Thus, it is shown that the developed carrier effectively releases the active substance into the wound surface, while the application time and concentration allow you to adjust the amount of its entry into the tissue.

table 2 Normalized fluorescence in organs and tissues of mice after application of zinc polycholinylphthalocyanine in the composition of the films according to example 7 Organs and Tissues gel film 0.02% according to example 4 gel film 0.2% according to example 5 The control 1 hour 2 hours 1 hour 2 hours Wound 10.6 ± 0.8 18.5 ± 5.0 > 38 > 39 1.2 ± 0.1 Skin adjacent 2.7 ± 0.5 3.5 ± 0.9 3.1 ± 0.8 4.0 ± 0.8 1.1 ± 0.1 Muscle adjacent 2.1 ± 0.7 3.0 ± 1.0 2.2 ± 0.6 3.0 ± 0.8 1.2 ± 0.1 Muscle distant 1.2 ± 0.1 1.2 ± 0.1 1.3 ± 0.1 1.2 ± 0.1 1.2 ± 0.1 Distant skin 1.0 ± 0.2 1.1 ± 0.1 1.3 ± 0.1 1.3 ± 0.2 1.1 ± 0.1 Liver 5.4 ± 0.3 4.7 ± 0.4 5.0 ± 0.4 4.8 ± 0.1 5.2 ± 0.6

Example 9. Determination of the rate of release of zinc polycholinylphthalocyanine when applying the inventive gel to a surface wound

The study was conducted in mice according to the method of example 8, applying 0.05 ml of gel to the wound instead of the film of example 2. As can be seen from table 3, the accumulation of the photosensitizer in the wound in the case of using a gel containing 0.2% zinc polycholinylphthalocyanine after 15 minutes reaches a significant value, while in remote areas of the skin and muscles it is found in small quantities. In the liver, with an increase in the time of application, F _n increases, in the kidneys the drug is registered in trace amounts, and in the spleen only after 6 hours.

Table 3 Normalized fluorescence in the organs and tissues of mice after application of the gel of example 2 with a content of 0.2% zinc polychinylphthalocyanine on the surface wound Organs Application time The control 15 minutes 2 hours 4 hours 6 o'clock Surface wound 68.3 ± 6.1 95.1 ± 3.6 93.2 ± 3.2 93.8 ± 4.5 2.5 ± 0.2 Surrounding skin 2.3 ± 0.6 5.6 ± 0.9 7.7 ± 1.1 7.5 ± 07 2.0 ± 0.2 Surrounding muscle 3.2 ± 0.9 11.1 ± 1.1 10.8 ± 1.6 9.9 ± 2.7 2.5 ± 0.2 Skin removed 1.7 ± 0.3 3.3 ± 0.7 4.1 ± 0.5 2.7 ± 0.8 2.0 ± 0.2 Remote muscle 2.3 ± 0.2 3.5 ± 0.4 3.8 ± 0.2 3.0 ± 0.3 2.5 ± 0.2 Fat tissue 2.3 ± 0.5 2.8 ± 0.4 3.2 ± 0.3 3.0 ± 0.6 2.2 ± 0.3 Liver 7.9 ± 1.8 10.9 ± 2.0 11.0 ± 1.1 12.0 ± 0.7 5.9 ± 0.7 Kidney 5.4 ± 0.3 5.9 ± 0.6 5.7 ± 0.9 5.8 ± 0.8 4.3 ± 0.4 Spleen 9.2 ± 2.3 9.1 ± 1.2 8.9 ± 0.8 14.3 ± 1.8 9.1 ± 0.5

Example 10. Determination of the release rate of zinc polycholinylphthalocyanine when applying the inventive gel to a deep wound

The study was carried out analogously to example 9. The drug was applied to the wound obtained by removing not only the skin, but also partially the muscles. As can be seen from table 4, the accumulation of zinc polycholinylphthalocyanian in a deep wound is very fast, while in remote areas of the muscle and skin, the drug is found in small quantities. In the liver and kidneys, with an increase in the time of application, F _n increases, and in the spleen the drug is registered only after 6 hours.

Table 4 Normalized fluorescence in organs and tissues of mice after application of the gel of example 2 with a content of 0.2% zinc polycholinylphthalocyanine on a deep wound Organs Application time The control 15 minutes 2 hours 4 hours 6 o'clock Wound > 100 > 100 > 100 > 100 2.5 ± 0.2 Surrounding skin 2.5 ± 0.2 6.6 ± 1.8 7.7 ± 2.2 7.6 ± 1.5 2.0 ± 0.2 Surrounding muscle 4.2 ± 0.5 12.4 ± 1.7 12.4 ± 1.2 10.6 ± 2.5 2.5 ± 0.2 Skin removed 2.2 ± 0.2 3.4 ± 0.6 4.0 ± 0.5 3.2 ± 0.6 2.0 ± 0.2 Remote muscle 2.6 ± 0.3 3.6 ± 0.5 3.9 ± 0.5 3.0 ± 0.5 2.5 ± 0.2 Fat tissue 2.2 ± 0.5 3.1 ± 0.5 3.5 ± 0.5 3.1 ± 0.5 2.2 ± 0.3 Liver 8.3 ± 0.9 10.6 ± 1.7 11.0 ± 1.8 12.8 ± 2.1 5.9 ± 0.7 Kidney 5.8 ± 0.5 5.7 ± 1.0 5.7 ± 1.0 6.2 ± 1.0 4.3 ± 0.4 Spleen 9.3 ± 2.9 9.0 ± 1.4 9.3 ± 1.8 14.5 ± 2.2 9.1 ± 0.5

Example 11

Patient A., 54 years old. Diagnosis "Non-healing wound of the left lower leg against type I diabetes mellitus". Objectively: on the left lower leg there is a purulent wound with an area of 8 cm ² . A gel obtained according to the procedure of Example 2 (exposure time 1 hour) was applied to the surface of the purulent wound, and laser irradiation was carried out daily for 15 minutes. For complete suppression of the purulent process complicated by type I diabetes mellitus, 6 AFDT procedures were required.

When exposed to AFDT, a quick cleansing of the wound from purulent necrotic masses was noted. Significantly reduced the appearance of granulation and epithelialization of the wound. When studying local microcirculation in the area of a non-healing wound, a decrease in edema and an improvement in blood flow in microvessels were noted. The next day, necrotic masses were actively rejected, there was practically no purulent discharge from the wound. Already on the 3rd day after the completion of the AFDT sessions, almost complete cleansing of the wound surface from purulent-necrotic exudate was noted, which was confirmed by clinical observations and cytological examination data. The size of the wound was reduced due to epithelialization and contraction of scar tissue. It should be noted that in smears from the surface of the wound after the first AFDT session, the number of neutrophils significantly decreased, which is probably due to the accumulation of a photosensitizer in them and their subsequent destruction under the influence of laser radiation. By the 7th day, marginal growth of immature epithelial cells was noted, and by the 14th day a mature epithelial layer was formed with complete healing of the wound.

Example 12

Patient B., 77 years old. The diagnosis of postthrombophlebitis disease. Trophic ulcer of the back and outside of the left foot. ” Objectively: on the outer and back side of the left foot there are two foci of trophic ulcers with an area of 3 and 5 cm ² , respectively. The gel obtained in example 2 was applied directly to the wound surface, covered with wax paper and a lightproof dressing. The gel exposure time was 2 hours. Laser irradiation sessions were carried out immediately after application of the gel. The radiation energy density is 60 J / cm ² , the irradiation time is 7 min per zone. Conducted 4 treatment sessions with an interval of 24 hours

Treatment has reduced the time for all phases of ulcer healing. Complete epithelialization was observed 21 days after the start of treatment.

Thus, the developed tool is convenient to use, ensures the penetration of the photosensitizer into the treated tissue at an economical consumption of the drug and complete healing of wounds in a minimum time.

Claims (2)

1. The tool for the treatment of purulent wounds by the method of photodynamic therapy, including zinc polycholinylphthalocyanine and water, characterized in that it is a gel of the following composition, wt.%:
zinc polycholinylphthalocyanine 0.02-1.00 hydroxyethyl cellulose 1.3-1.6 dimethyl sulfoxide or benzalkonium chloride (alkylbenzyldimethylammonium chloride) 0.2-3.0 block copolymer of ethylene oxide and propylene oxide or polyethylene glycol 3.2-4.0 propylene glycol 4.0-7.7 water rest 2. An agent for treating purulent wounds by photodynamic therapy, including zinc polycholinylphthalocyanine and water, characterized in that it is a film obtained by drying the gel according to claim 1.