RU2186563C2 - Preparation for treating a burn wound - Google Patents

Abstract

FIELD: medicine, pharmacology. SUBSTANCE: preparation is a mixture of fatty alcohols at 5-100% total content of the following alcohols: octadecatetraenic, eicosapentaenic, docosapentaenic and docosahexaenic. The usage of such a preparation one should reveal improved inhibition of inflammatory processes that decreases the severity of damage and creates conditions for both lower volumes of necessary reparation and its stimulation, as well. As a result, quicker detachment of scabs is observed at earlier stages of their development along with complete wound healing at no complications and associated side phenomena. EFFECT: broadened number of preparations. 1 ex, 1 tbl, 6 dwg

Description

 The invention relates to medicines containing organic active ingredients, namely higher fatty alcohols.

 In Russia, the following drugs and groups of drugs are used as anti-burn agents: sulfanilamide preparations (streptocide, etc.), antiseptic agents (potassium permanganate, hydrogen peroxide solution, etc.), biogenic stimulants (aloe liniment, Kalanchoe juice), vinylin, linoleum, linz of tezan, tannin, enzymatic preparations (trypsin, chymotrypsin, etc.), as well as antibiotics, anabolic substances, vitamins and a number of others (Mashkovsky I.D. Medicines. - Kharkov: Torsing, 1997. - T. 1 and 2) .

 Of the aliphatic compounds that are similar in structure (but not identical), omega-3 marine fatty acids have a pronounced biological effect (Kinsella JE, Lokesh B., Stone RA Dietary n-3 polyunsaturated fatty acids and amelioration of cardiovascular disease: possible mechanisms // Am. J. Clin. Nutr. - 1990. - Vol. 52, N 1. - P.1-28), including for burn injuries (Pat. 1805961 of the Russian Federation, MKI A 61 K 31/20. Means showing anti-burn activity / Latyshev N.A., Kasyanov S.P., Dobrynchenko I.V., Kirillov O.I. - 1993).

 It is known to use a mixture of higher fatty alcohols in detergents (shampoos, soaps, etc.) as surfactants (Pletnev M. Yu. Cosmetic and hygienic detergents. - M .: Chemistry, 1990. - 272 p. )

 To date, it is not known the use of agents containing higher fatty alcohols omega-3 series in the treatment of burn wounds.

 In order to expand the arsenal of means of influencing a living organism, higher omega-3 fatty alcohols are used as an anti-burn agent in the present invention.

 The use of omega-3 fatty alcohols in case of burn injury gave an unexpected result in qualitative and quantitative characteristics, superior to the use of the commercial anti-burn drug “Linetol” (a preparation based on fatty acids of flax oil, contains up to 55% - linolenic acid - omega-3 fatty acid series of plant origin).

 Linetol, as a drug, was chosen as a comparison when testing the effectiveness of the action of omega-3 alcohols, as the official drug closest in chemical structure recommended by the Pharmacopoeia for use in these pathologies.

 Mixtures of fatty alcohols, including omega-3 alcohols, are an oily mobile liquid of a slightly yellowish color (to colorless) with a peculiar soft smell. The mixture does not lose its liquid consistency even at low temperatures; not combustible and does not contain any solvents. Hereinafter, the mixture of higher fatty alcohols of the omega-3 series will be referred to as a product containing omega-3 fatty alcohols.

Препарат стандартизировали по сумме четырех омега-3 спиртов: (1) октадекатетраеновый спирт, (2) эйкозапентаеновый спирт, (3) докозапентаеновый спирт, (4) докозагексаеновый спирт. Ниже приведены их химические формулы:

цис-6,9,12,15-октадекатетраеновый спирт (спирт 18:4ω3)

цис-5,8,11,14,17-эйкозапентаеновый спирт (спирт 20:5ω3)

цис-7,10,13,16,19-докозапентаеновый спирт (спирт 22:5ω3)

цис-4,7,10,13,16,19-докозагексаеновый спирт (спирт 22:6ω3)
Омега-3 или ω3 обозначает расположение первой двойной связи в молекуле от концевой метильной группы. Остальные связи упорядочены и располагаются через метиленовые мостики. Под названием каждого из спиртов (по системе IUPAC) приводится его сокращенная формула: первая цифра обозначает количество атомов углерода, вторая - двойных связей.In nature, omega-3 fatty alcohols are found mainly in marine organisms (fish, plankton, etc.). Terrestrial animals and plants are, in this case, a rare exception. But even in marine objects, the concentrations of these substances are insignificant; as a result, it seems difficult to isolate any significant quantities of these alcohols. Therefore, in our work, we prepared them directly from omega-3 series marine polyenoic fatty acids, which were obtained by known methods (Pat. 1581737 RF, MKI C 11 C 1/00, C 11 B 7/00. A method for producing a concentrate of ethyl esters eicosapentaenoic and docosahexaenoic acids / Latyshev I. A., Kasyanov S. P., Glushenko T. V. - 1993; Pat. .A., Imbs A.B., Gaidai N.V., Kuklev D.V., Kasyanov S.P. - 1993). The resulting fatty acids were reduced to alcohols by the standard procedure (Carrer P. Course in Organic Chemistry. - L .: State Scientific and Technical Publishing House of Chemical Literature, 1960. - P.109-113). General scheme for obtaining:

The drug was standardized by the sum of four omega-3 alcohols: (1) octadecatetraenoic alcohol, (2) eicosapentaenoic alcohol, (3) docosapentaenoic alcohol, (4) docosahexaenoic alcohol. Below are their chemical formulas:

cis-6,9,12,15-octadecatetetraene alcohol (alcohol 18: 4ω3)

cis-5,8,11,14,17-eicosapentaenoic alcohol (alcohol 20: 5ω3)

cis-7,10,13,16,19-docosapentaenoic alcohol (alcohol 22: 5ω3)

cis-4,7,10,13,16,19-docosahexaenoic alcohol (alcohol 22: 6ω3)
Omega-3 or ω3 denotes the location of the first double bond in the molecule from the terminal methyl group. The remaining bonds are ordered and located through methylene bridges. Under the name of each of the alcohols (according to the IUPAC system) its abbreviated formula is given: the first digit indicates the number of carbon atoms, the second - double bonds.

 The content in the preparation of an effective beginning - the amount of these alcohols can range from 5 to 100%. The lower limit is determined on the basis that a decrease in the total content of these alcohols below 5% significantly or completely reduces the effectiveness of the product due to the high "ballast" content of other fatty alcohols that do not have such an effect. The upper limit corresponds to the moment when the tool practically contains only the indicated alcohols. Such a tool has the greatest activity. However, due to certain difficulties in obtaining highly enriched concentrates of omega-3 acids, from which the corresponding alcohols are prepared, their concentration in a ready-to-use agent is, on average, 20-40%, while the ratio of alcohols in the mixture does not matter, since each of them possesses anti-burn activity comparable to others, and only their total amount equal to 5-100% is important.

 Under the influence of a thermal stimulus, a multiphase inflammatory reaction develops (Chernukh A.M., Frolov E.P. Kozha. - M .: Medicine, 1982. - 336 p.) And according to the time of occurrence of some morphological elements and their successive change to others, an opportunity to judge the dynamics of inflammatory phenomena in the lesion.

 Thus, the structure of the skin of rats is basically similar to human and also represents a three-component system formed by the epidermis, dermis and subcutaneous fatty tissue, most pronounced in the back (Gambaryan P.P., Dukelskaya N.M. Rat. - M. : Soviet science, 1955. - 197 p.), But the epidermis of the vast majority of the skin surface, with thickly growing hair, is less complex and, instead of five layers in humans, in animals consists of only three: basal, prickly and horny (Ivanov I. .F., Kovalsky P.A. Cytology, histology, embryo iologiya. - M .: Kolos, 1976.- 448 p.). In this regard, during the evolution of a burn wound in rats, the formation of such a very typical morphological element as a bubble is not observed. In addition, the Wistar rats used in our experiments are albinos in which the specific function of melanocytes located in the basal layer of the epidermis is impaired, as a result of which they are not able to synthesize melanin pigment. Nevertheless, in these animals, the area and especially the depth of skin lesions, like human skin, depend both on the parameters of the heat source (size, temperature, heat capacity, duration of exposure), and on the moment of active therapy, its effectiveness in counteracting the development of the inflammatory reaction. Moreover, the severity of pathological changes decreases in the direction from the surface of the skin to its deeper layers and from the center of the burn area to the periphery.

 Externally, the zone of coagulation necrosis immediately after exposure to the temperature stimulus chosen by us instantly turns pale, and then stands out with its more pink color from the surrounding intact skin. After a day, as a result of a two-phase reaction of the preserved microvasculature, leading to the expansion of small vessels, their permeability increases, which is accompanied by the release of the liquid part of the blood, its formed elements and protein into the intercellular space (Chernukh A.M. Inflammation. - M .: Medicine , 1979.- 447 p.).

 The erythrocytes that came out in this way (by diapedesis) are hemolized, which is very typical for burns, and the hemoglobin released due to this, is subsequently converted into a series of products, turns into hemosiderin. It forms during the first 24 hours from the time of the hemorrhage, which gives the tissues a reddish-brown color (Vikhreev B.S., Burmistrov V.M. Ozhogi. - L .: Medicine, 1986. - 272 p.). On the surface of the skin, this manifests itself in the form of a primary morphological element - a spot. The spots are numerous, irregular in shape and tend to gradually merge with each other, and the accumulation of hemosiderin leads to the formation of a secondary morphological element - brownish hyperpigmentation (Vladimirov V.V., Zudin B.I. Skin and venereal diseases. Atlas. - M .: Medicine , 1980. - P. 15-36). Since, in our experiments, albino rats were used, in which, as indicated above. the formation of melanin pigment is impaired, it is more likely to say that it is hemosiderin that determines the color of the formed new pathological element.

 In parallel, the exit of the liquid part of the blood with an extensive set of proteins leads to the impregnation of the surrounding tissues, and the evaporation of moisture from the surface of the burn enhanced by 16-20 times compared with healthy skin (B. Vikhreev, V. M. Burmistrov Burns. - L .: Medicine, 1986. - 272 p.) Causes the drying of the resulting necrotic mass and the formation of the next pathological element - a light brown scab. The scab protrudes above the surface of healthy skin, is elastic upon palpation.

 Damage to the endothelium of the blood vessel initiates the process of hemostasis. The vital activity of cell populations ceases and the necrotic zone only slightly expands, and mainly deepens its size. The stasis zone moves deeper into the hyperemia zone and new larger blood vessels are involved in the exudation process. The clinical manifestation of this is a significant darkening and thickening of the scab, which remains elastic. The appearance of such a dark red scab on human skin occurs with burns of the IIIa degree, and a darker, dry and dense scab (in rats it resembles a coin), with a pattern of dilated blood vessels in a stasis state and often fused with the underlying tissues corresponds to IIIb degree ( Dolinin V.A. Thermal lesions.In the book: Field Surgery / Edited by Lisitsin K.M., Shaposhnikov Yu.G. - M .: Medicine, 1982 - S. 153-190). Theoretical extrapolation of the pathological anatomy of third-degree burns of human skin of rats to the skin of rats suggests that the latter have lesions of the skin integument throughout its thickness, including subcutaneous fatty tissue.

 At the same time, the formation of such a dense dark brown scab, although fraught with a secondary violation of microcirculation in adjacent tissues, in albino rats marks the actual completion of pathological changes and the transition to reparative processes. In principle, the termination of the destructive process is possible at any stage, but practically no scab formation was observed before. This is preceded by a slowdown in the evolution of a burn wound, that is, a stop on a certain pathological element, without the appearance of the next one to two days. Then, along the periphery of the scab, its edge is detached from the underlying skin in the form of a narrow strip, which gradually expands towards the center of the scab until it is completely rejected.

 The final resolution of the wound was considered to be its condition in which a clean skin surface appeared under the scab without visually detectable differences with intact skin. But in some animals, burn complications were observed, manifested in the appearance of secondary wounds and scabs, suppuration, a significant thickening of the skin in the area of injury, as well as the formation of hypertrophic scars, changes in skin color (pallor, whitish areas, redness) and / or skin atrophy phenomena (violation normal skin pattern, thinned, sagging areas) in place of the fallen scab.

Ultimately, the four most typical and consecutive stages in the studied clinical picture were taken as defining landmarks:
I - the skin is elastic throughout the entire thickness, there are spots due to hemolysis and the formation of hemosiderin;
II - the skin is elastic throughout the entire thickness, together with the soft light brown scab available on its surface;
III - the entire thickness of the skin is elastic together with the red-brown scab on its surface;
IV - a dense, coin-like, dark brown scab, often with a different pattern of dilated blood vessels in a stasis state.

 To exclude intermediate stages during the transition of stages and for randomization of observations according to Avtandilov (G. Avtandilov, Medical morphometry. Guide. - M .: Medicine, 1990. - 384 p.), We used the transparent grid proposed by him with equidistant test points, according to which we measured on the affected skin surface the areas occupied by successive pathological elements successively replacing each other. With the predominance of one of them, an animal with such a wound was assigned to the corresponding stage. According to the total data, histograms were constructed where the size of each column was determined by the number of rats having wounds at a particular stage of evolution, and the number of columns corresponded to the presence of specific stages in this experimental group. Rats were examined daily and continued to redistribute in the usual manner as their wounds evolved. For clarity, stage IV is highlighted as the most unfavorable.

где Χ² - критерий соответствия (критерий согласия);
f₁ и f₂ - частоты классов первого и второго рядов;
n₁ и n₂ - число особей в каждом из них, с последующим определением величин вероятности по номограмме критерия Χ² и тесту совпадения значения Χ². Статистически достоверными считали различия при Р≤0,05.Statistical processing of rat distribution histograms was carried out using the consent criterion Χ ² (Rokitsky PF Basics of variation statistics for biologists. - Minsk: Publishing house of the Belarusian State University, 1961. - 222 p.) According to the formula:

where Χ ² - criterion of compliance (criterion of agreement);
f ₁ and f ₂ are the frequencies of the classes of the first and second rows;
n ₁ and n ₂ are the number of individuals in each of them, with the subsequent determination of the probability values from the nomogram of the criterion Χ ² and the test of coincidence of the value Χ ² . The differences were considered statistically significant at P≤0.05.

 Example 1. The treatment for burn wounds, including omega-3 fatty alcohols.

 In the experiment, 126 female Wistar rats (from the Stolbovaya RAMS cattery) weighing 130-150 g were used, which were divided into 6 groups of 21 animals each. Rats of all groups were in cages of 7 individuals. The animals were kept for the entire period on a standard diet adopted in the nursery, water ad libitum.

Group 1 consisted of untreated rats; it served as a control for the 3rd (Lynetol) and 6th (ballast alcohols) groups;
The 2nd group of rats after the burn was treated with medical vaseline, the rats of this group served as a control for the 4th (omega-3 alcohols, 59%) and 5th (omega-3 alcohols, 100%) groups;
Group 3 was treated with a commercial anti-burn agent Lynetol;
Group 4 was treated with a test agent omega-3 alcohols of the following composition,%:
Octadecatetraenoic alcohol - 7
Eicosapentaenoic Alcohol - 25
Docosapentaenoic alcohol - 5
Docosahexaenoic Alcohol - 22
The total total content is 59%. The remaining fatty alcohols, as minor components, accounted for 41%.

The 5th group was treated with the test agent omega-3 alcohols of the following composition,%:
Octadecatetraenoic alcohol - 12
Eicosapentaenoic Alcohol - 35
Docosapentaenoic alcohol - 9
Docosahexaenoic alcohol - 44
The total total content is 100%. The remaining fatty alcohols were absent.

 This group 5, like the next 6, was taken in an experiment to determine the effective concentration of omega-3 alcohols in the preparation.

6 group was treated with a mixture of "ballast" fatty alcohols with a low content of active omega-3 alcohols of the following composition,%:
Octadecatetraenyl alcohol - 0.5
Eicosapentaenoic Alcohol - 1.3
Docosapentaenoic alcohol - 0.3
Docosahexaene alcohol - 2.9
The total total content of omega-3 alcohols is 5%. The remaining "ballast" fatty alcohols accounted for 95%.

Before the experiment, under ether anesthesia in rats, wool was removed in the middle third of the back from a 25–25 mm skin area. The burn was caused within 3 seconds by applying a massive (500 g) metal object to the skin, which had a round, even surface with a diameter of 20 mm at a temperature of 80 ^o C. at the contact point. To obtain a superficial thermal injury, immediately before the burn, a syringe was introduced under dehydrated skin until 3 cm ^{3 of} air to form a uniform air cushion at the contact point. This manipulation made it possible to ensure complete and uniform contact with a physical agent - a source of excess heat. Thus, only the skin with all its appendages and systems was exposed to temperature effects, and the underlying tissues remained almost intact. After removing the injection needle, the introduced air quickly escaped and the “pillow” subsided. After a burn, 150 mg per test wound was applied to the affected skin surface at the test temperature. Until complete absorption of the preparations, the rats were kept separately from each other in order to avoid mutual licking of the wounds. The experiment was carried out from 9 to 12 hours, the preparations were repeated at 18-19 hours. On the following days, the treatment time remained the same. At the same time, the time of detection at the injury site, visually and by palpation of specific objective changes in the skin - the so-called morphological elements, which are an external manifestation of the corresponding pathological processes in the skin, was recorded.

 The survey results are shown in digital material in the table, as well as for better visualization in the form of histograms in figures 1-6. Stage IV is shaded on the histograms as the most unfavorable.

 The results of the studies demonstrated the effective local effect of agents, including the higher omega-3 fatty alcohols in thermal skin lesions. When using these funds, there was a clear inhibition of inflammatory processes, which reduced the severity and created conditions for both a decrease in the volume of necessary repair and its stimulation. As a result, everything was expressed in the soon rejection of scabs at the earlier stages of their formation, with complete healing of the wounds without complications and associated side effects. Thus, omega-3 fatty alcohols must be considered as highly effective agents for topical treatment of burn lesions.

Claims (1)

 The treatment for burn wounds, characterized in that it is a mixture of fatty alcohols, including 5-100% - higher omega-3 fatty alcohols with a chain length of 18-22 carbon atoms and 4-6 double bonds.

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