KR101168981B1 - Composition of therapeutical agents for a burn containing Citric acid, Zincand L-Arginine - Google Patents

Abstract

The present invention is to remove the poison by verifying the healing effect of the skin coating composition for treating burns containing test substance citric acid, zinc and L-arginine for burns by surgical treatment using rats of the Sprague-Dawley strain At the same time, the present invention relates to a composition for treating burns comprising citric acid, zinc and L-arginine, which can be used as a burn treatment agent capable of treating burns by assisting skin regeneration.

Description

Composition of therapeutical agents for a burn containing Citric acid, Zincand L-Arginine}

The present invention relates to a composition for treating burns comprising citric acid, zinc, and L-arginine, and more particularly, to a test material cisar concentrate and cigar for burns by surgical treatment using rats of the Sprague-Dawley strain. To verify the healing effect of the skin application of Le Coniferous Stock Solution B and to provide a composition for the treatment of burns comprising citric acid, zinc and L-arginine which can be used as a burn treatment agent.

Skin is the largest organ and complex organ in the body, acting more than a physical defense against infection and dehydration. It also helps regulate body temperature by controlling blood flow and evaporation of sweat, and it is also a very important sensory organ.

The thickness of the skin varies with age and body parts, but on average, it ranges from 1mm to 2mm and has two layers, regardless of thickness. The outer thin epidermis receives nutrition from the thicker and more sensitive dermis located inside it. The outer surface of the epidermis is made up of dead cells. When these cells fall off, they are replaced by new cells that grow underneath. The innermost part of the epidermis is made up of rapidly dividing cells, called keratinocytes, which produce tough proteins called keratin, which also contain unique fatty substances that make the skin waterproof. have.

Blood vessels, lymphatic vessels and nerves in the skin are in the dermis. Hair follicles, sweat glands, and oil glands are also deep in this layer, mostly connective tissue. Collagen, the most common protein in the body, gives skin flexibility and support structure, and is mainly in the form of fibroblasts. The dermis acts to prevent wound shrinkage and scarring.

Images can be classified as follows depending on the cause.

It refers to a case where a burn is caused by a fire or intense heat due to a fire accident or an explosion of propane gas or LPG gas, and it is usually caused by a flame burn that is deep and may cause damage to the respiratory tract, and hot water, cooking oil, steam, etc. In most cases, burns are more than 2 degrees, and hot water burns in children and electric shocks can cause burns even at low voltage used in homes, often causing severe sequelae. It is a burn caused by contact with an electric burn, acid, alkali, or general organic solvent, which can cause serious obstacles in some cases, and it can be worn when exposed to strong sun in summer, and sunscreen is applied to the skin. Sunburn, hot iron plate, iron, all For the most part as an image generated as the skin is exposed to contact pad is divided into images that can be conducted with more than three degree burns.

The degree of burn is often divided into three to four stages ranging from the first to the third degree or the fourth degree according to the symptoms of the burn, and the first degree burn is accompanied by pain such as redness of the skin and tingling. It causes damage to the epidermis, the outermost of the skin layers, and swells with pain and erythema. The symptoms disappear in a few days, but there may be a slight fallout and pigmentation in place. No scars remain after recovery. Sun burn is an example of the most common first degree burns.

Second-degree burns also affect both the epidermis and dermis, causing erythema, pain, edema, and blistering within 24 hours of the accident. This burn also affects the gland and pores. Awakening is burning and painful. When the blisters burst, they show erosions and a large amount of secretion. Special care should be taken when the burned area reaches more than about 15-30% of the body surface area. Heals within a few weeks, but pigmentation and discoloration often remain in place. The secondary infection causes more local symptoms and lasts longer. Third-degree burns affect the epidermis, dermis, and hypodermis, resulting in darkening or translucent whitening of the skin and clotting of blood vessels just below the surface of the skin. The burn may become numb, but the patient feels extreme pain, necrosis of the skin tissue and structure, which takes a lot of time to heal and scars. Two weeks after the accident, the scab peels off and an ulcer surface appears. There is a lot of secretion and easy to bleed, but gradually new tissue develops, the epidermis regenerates, leaving scars and healing. In cases of deep skin necrosis or secondary infections, healing may be delayed and the surface of the scar may become irregular, resulting in keloids or deformations or movement disorders. Special care should be taken when the burned area covers more than 10% of the body surface area. In addition, a 4th degree burn is a case where the burnt part tissue carbonized and turned black, and is a severe case of a 3rd degree burn. Burn therapy is a remedy for regenerating skin tissue within a short period of time without leaving asymptomatic as possible, and care should be taken not to cause bacterial infection.

Commercially available burn treatments include formulations based on fucidin acid or sodium fucidate (trade name: fucidin ointment, cream, gel type), formulations containing silver sulfadiazine as the main component (trade name: thread margin), β- Formulation containing two major components (Crecord): Cytosterol as a main ingredient (trade name: Unbottled Ointment), Calcipotriol (trade name: Divonex), Gentamicin and Betamethasone Valerate, The formulation (brand name: carbate) etc. which contain prednica carbate as a main component are marketed.

Applicant disclosed in Patent Invention No. 0428587 containing a composition comprising a citric acid, zinc and L-arginine together with a pharmaceutically acceptable carrier, while the active ingredient of citric acid, zinc and L-arginine is effective in killing cancer cells. In addition, even users of existing anticancer drugs have been able to use anticancer drugs as a factor to drastically reduce the problem of normal cell destruction.

Accordingly, the present inventors have tried to improve the present invention by confirming that a composition containing citric acid, zinc, and L-arginine has been studied for burn treatment along with the burn treatment agent of the present invention. do.

The present invention has been made to solve the above problems, its purpose is to treat burns containing test substance citric acid, zinc and L-arginine for burns by surgical treatment using rats of the Sprague-Dawley strain The present invention provides a composition for treating burns, including citric acid, zinc, and L-arginine, which can be used as a burn treatment agent by verifying the healing effect of skin coating.

According to a first aspect of the present invention for achieving the above object, there is provided a composition for the treatment of burns containing citric acid, zinc and L-arginine as active ingredients in a therapeutically effective amount together with a pharmaceutically acceptable carrier and diluent. .

At this time, according to an additional feature of the present invention, it is preferable that the active ingredient contains 0.02 to 20 wt% citric acid, 0.0001 to 1.0 wt% zinc and 0.025 to 25 wt% L-arginine as the total weight of the composition. .

In addition, according to an additional feature of the present invention, it is preferred to combine the composition with a pharmaceutically acceptable ointment.

In addition, according to an additional feature of the present invention, the ointment is preferably selected from petrolatum, paraffin, vegetable oil, lard, wax, and sweet ointment as an oil-based base.

The composition for the treatment of burns comprising citric acid, zinc and L-arginine according to the present invention is a test material for the burn by surgical treatment using rats of the Sprague-Dawley strain sterilized with citric acid, zinc and L-arginine 100 The composition for the treatment of burns by mixing the% composition amount can be used as a burn treatment agent that can help not only burn treatment but also human life by eliminating poisoning and assisting skin regeneration by verifying the healing effect by skin application.

In addition, even if the skin is not irritated at all and used for various methods such as application and adhesion to the soft skin, it does not cause skin trouble, and even if the burn treatment is completed by assisting the regeneration of damaged skin tissue or other muscle tissues at the same time as it quickly repairs the burn. It can be used as a burn treatment to remove the remaining burns.

1 is an image model of the czar concentrate according to the present invention (image section length)
Figures 2a to 2e is a three-week skin coating burn treatment numerical table of the burn length
Figure 3 is an image model (image length vertical) of the czar concentrate according to the present invention
Figures 4a to 4e is a three-week skin application burn treatment numerical table of the length of the burn
Fig. 5 is an image model of a czar concentrate according to the present invention (image front part)
6a to 6e is a three-week skin application burn treatment numerical table of the burner front part
FIG. 7 is a graph depicting histopathological records of cisar conjugates according to the present invention.
8a to 8e is a burn table according to histopathological records
Figure 9 is a diagram showing the histopathological state of the untreated group (G1) an embodiment of the present invention.
Figure 10 is a diagram showing the histopathological state of concentrate (G3) that is one embodiment of the present invention.
Figure 11 is a diagram showing the histopathological state of concentrate undiluted solution B (G4) of one embodiment of the present invention.
Figure 12 is a diagram showing the histopathological state of the positive control group (G5) of one embodiment of the present invention.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

The present invention relates to citric acid, zinc and L as a linked invention of an anticancer composition containing the applicant's patent invention No. 0925516 containing zinc and L-arginine and an anticancer composition comprising the citric acid, zinc and L-arginine of Patent Invention 0428587. Using a composition comprising each active ingredient of citric acid, zinc and L-arginine to provide an arginine burn agent composition through a comparative experiment group through animal experiments (mouse experiment) to present the treatment and effects of burns As a feature of the burner composition, a test article containing the active ingredients of citric acid, zinc and L-arginine of the present invention is named "cisar concentrate (G3)" to describe the technical part and efficacy in detail. It was.

Citric acid may be included in the pharmaceutical composition according to the present invention at 0.02% to 20% by weight, preferably 0.01% to 1.0% by weight, most preferably 0.02% to 0.2% by weight, based on the total weight of the composition.

Citric acid used in the present invention may be included by extracting citric acid present in the free state in the seeds or juice of various plants. Citric acid is also prepared by the surface fermentation process or submerged fermentation process using the fungus Aspergillus niger, which can also be used in the composition of the present invention. In the production process of citric acid, molasses is used as a sugar-containing raw material and various centrifuges such as nozzle type centrifuge, automatic discharge type centrifuge, and decanta are used.

Zinc in the pharmaceutical composition according to the present invention may be included at 0.001% to 1.0% by weight, preferably 0.0005% to 0.05% by weight, most preferably 0.001% to 0.01% by weight, based on the total weight of the composition.

Zinc is present in traces in our diet. In particular, zinc is found in sufficient concentrations in animal foods such as oysters, crustaceans, fish, and red meat, as well as in various plant foods such as grains, beans, nuts, and seeds.

L-arginine in the pharmaceutical composition according to the present invention may be included at 0.025% to 25% by weight, preferably 0.0125% to 1.25% by weight, most preferably 0.025% to 5.0% by weight, based on the total weight of the composition. .

L-arginine exists as one of the amino acids that make up a protein, which is a non-essential amino acid for adults but an essential amino acid for infants. It belongs to the protein protamine in fish sperm, and in herring, salmon, etc., about 70% of the constituent amino acids are arginine. L-arginine also exists freely in plant seeds. As a metabolic pathway in vivo, it is a constituent of the ornithine cycle discovered by HA crebes and the like, and is broken down into urea and ornithine by the action of arginase and is produced from citrulline and aspartic acid in vivo. L-arginine has a function of protecting the human body from the poisoning effects of ammonia and large amounts of amino acids.

Carriers used in the pharmaceutical compositions of the present invention include carriers, adjuvants and vehicles commonly used in the pharmaceutical art and are collectively referred to as “pharmaceutically acceptable carriers”. Pharmaceutically acceptable carriers that may be used in the pharmaceutical compositions of the invention include, but are not limited to, ion exchange, alumina, aluminum stearate, lecithin, serum proteins (eg, human serum albumin), buffer materials (eg, Various phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids), water, salts or electrolytes (e.g. protamine sulfate, disodium hydrogen phosphate, hydrogen carbonate, sodium chloride and zinc salts) Silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substrates, polyethylene glycols, sodium carboxymethylcellulose, polyarylates, waxes, polyethylene-polyoxypropylene-blocking polymers, polyethylene glycols and wool, and the like.

The pharmaceutical composition of the present invention may be administered via any general route as long as it can reach the desired tissue. Accordingly, the pharmaceutical compositions of the present invention may be administered topically, orally, parenterally, intraocularly, transdermally, rectally, intestine, and the like, and may be formulated into solutions, suspensions, tablets, pills, capsules, sustained release preparations and the like.

In one embodiment, when the pharmaceutical composition of the present invention is formulated as an oral solid, diluents (e.g., lactose, dextrose, sucrose, cellulose, corn starch or potato starch), active agents together with the active ingredient (E.g. silica, talc, stearic acid, magnesium or calcium stearate and / or polyethylene glycol), binders (e.g. starch, arabic gum, gelatin methylcellulose, carboxymethylcellulose or polyvinyl pyrrolidone), boron Release (e.g., starch, alginic acid, alginate or sodium starch glycolate), formal mixtures, dyes, sweeteners, wetting agents (e.g. lecithin, polysorbates, laurylsulfate) and generally used in pharmaceuticals It may contain pharmaceutically inert substances. These agents can be prepared by known methods, for example by means of mixing, granulating, tableting, dragging or film-coating processes.

The composition of citric acid, zinc and arginine according to the present invention is usually composed of a ratio of 20: 1: 25 and synthesized the composition and the additional material to carry out the experiment with the test substance cisar concentrate according to the present invention. .

The invention will be more specifically illustrated by the following examples and experimental examples. However, these examples and experimental examples are only embodiments of the present invention and do not limit the scope of the present invention.

≪ Example 1 >

Sizar Concentrate Tablets Manufacturing

In a suitable container, 30% by weight of citric acid, zinc and L-arginine (pH buffering) and lactose, 5% by weight of magnesium stearate, 10% by weight of sodium starch glycolate, and 100% of sterile water in a suitable container After mixing the amounts, it was stirred for 1 hour while maintaining at 30 to 60 ℃, cooled to room temperature and then compressed according to the conventional method for preparing a tablet to prepare a tablet containing 350mg each composition.

When the composition consisting of citric acid, zinc and L-arginine is named "cisar concentrate" according to the procedure described above, using a Sprague-Dawley rat, causing burns and applying cisar concentrate to the skin. The effect of healing and its efficacy will be presented in detail through the experimental group.

Test group configuration and setup

1.Test Group Configuration and Dose Settings

Sprague-Dawley strain rats according to the present invention was experimented by setting up five test groups to determine the healing effect by the skin application of cisar concentrate by causing burns. That is, it was classified into G1 untreated group, G2 sterilized injection water treatment group (negative control), G3 to G4 test substance treatment group, and G5 positive control group.

More specifically, as shown in the following table, the constituent dose of the test group was set to confirm the healing effect according to the burn of the present invention.

(1) Composition of test group

 Configuration group gender Number of animals Animal Number Applicable Material Coating amount (ml / head) G1 Male 10 1 to 10 - G2 Male 10 11 to 20 Sterile Injection Water 0.1 G3 Male 10 21-30   Outlet rate 0.1 G4 Male 10 31-40   Concentrate undiluted solution B 0.1 G5 Male 10 41-50 Margin 0.1

G1 ~ G5: Burn-induced group, G1: Untreated group, G2: Sterile injectable water treatment group (negative control), G3 ~ G4: Test substance treatment group, G5: Positive control group

(2) Test group selection

Rats used in the present invention are small in size, easy to handle, have low seasonal variability, are less susceptible to stress, and have a stronger resistance to disease. In addition, the response to the drug is constant, so there is little difference in sex response according to the male and female, and abundant test basis data are accumulated. It is widely used in safety pharmacology test because extrapolation of experimenter is possible.

(3) separation of test group

Among the animals determined to be healthy during the acclimation period, hair removal was performed on the day of group separation, and 50 animals of good skin condition were selected.

(4) administration

The skin coating was selected by the investigator as a clinical route, and the test substance was applied to the burn-inducing site at a dose of 0.1 ml / head for 1 time / day, 7 times / week, and 3 weeks.

(5) Observation and Inspection Items

 -Burns

(a) Ketamine and rumoons are mixed 4: 1 and intraperitoneally administered at 1 ml / kg, followed by anesthesia using a hair removal machine to remove a certain area (approximately 3.0 cm x 3.0 cm) on the back and finely using an electric razor. Epilation.

(b) After earning 70% alcohol on the epilated area, space the center of the vertebral line about 1 cm and place a 1.5 cm x 1.5 cm square plate attached to the electric iron on the back at about 80 ° C. Contact occurred for ˜ 20 seconds to cause burns.

(c) The test substance, the positive control material and the negative control material provided by the sponsor were applied to the burn-induced site.

-Observation and Inspection Items

(a) After the image induction, the size of the image was measured and photographed. Image area measurement and photographing were performed twice a week.

(b) The condition and death of the burned site were visually observed and photographed on the day of the measurement of the burned area.

(c) An autopsy was performed 21 days after administration, and histopathological specimens were made after fixing the administration site (skin) of all animals in 10% neutral buffered formalin solution.

(d) In histopathological examination, burns were examined for six items: epithelialization, inflammation, angiogenesis, fibroblast activation, collagen deposition and wound contraction.

-Test result display

The measurement results showed the mean ± standard error of the width, length, and area of the burned area. The histopathological results were divided into five stages.

(6) statistical method

The comparison between the test substance administration group and the positive control group administration group for the excipient control group was verified by one-way ANOVA.In this case, if the significance was recognized, the Duncan test was recognized if the equal dispersion was recognized. If not, a post test was performed using Dunnett's test. The significance of the significance was p <0.05 or less, and SPSS 10.1, a commercial statistics program, was used according to the SOP for statistical processing.

Experimental Example 1

According to the test group configuration and setting described above, the present invention induced a burn by surgical treatment on the rat, and then applied the test material for 3 weeks to verify the healing effect on the burn.

The test group consisted of the untreated group (G1), the negative control group (G2), the test substance cisar concentrate (G3), and the czar concentrate undiluted solution B (G4) administration group, and the positive control group with predominantly 1% cream margin ( G5). After burnout, burn length measurement and photographing were performed twice a week, and histopathological examination was performed on day 21 after administration. The test results are as follows.

Experiment Result 1

(1) As a result of observation of general symptoms according to the administration and test of the test substance, no systemic symptom change related to administration of the test substance was observed.

(2) As a result of measuring the length of the image and taking the photographs according to the above test, the width and length of the image were measured as shown in FIGS. 13 to 19, and the horizontal length was absent on the 16th and 20th days. The czar congestion rate (G3) was reduced compared to the treatment group (G1), and decreased at 20 days compared to the negative control group (G2). Sisar concentrate undiluted solution B (G4) had a smaller wound size than the untreated group (G1) and negative control group (G2) at 20 days.

On the other hand, the vertical length was smaller in the czar concentrate (G3) than in the untreated group (G1) and the negative control group (G2) on the 16th and 20th days, and the czar concentrate undiluted solution B (G4) was not treated on the 20th day. Decreased values than the group (G1).

That is, as a result of calculating the area using the measured horizontal and vertical values, the czar concentrate (G3) was smaller in size than the untreated group (G1) and the negative control group (G2) on the 16th and 20th days, and the concentrate undiluted solution B (G4) showed a value smaller than the no treatment group (G1) on the 20th day.

On the other hand, the positive control group (G5) was significantly larger than the untreated group (G1) and negative control group (G2) at the 16th and 20th days, and the wound healing rate was delayed.

3) As a result of histopathological examination, the degree of epithelialization was higher in the czar concentrate (G3) and the uncontained undiluted solution B (G4) than the untreated group (G1), and the wound contraction was negative control group (G1). G2) and concentrate undiluted solution B (G4) were increased.

In addition, the positive control group (G5) had significantly reduced inflammation compared to the untreated group (G1).

Based on the above results, it is thought that the czar concentrate (G3) and the czar concentrate undiluted solution B have an effect of reducing the size of the window surface and promoting the progression of epithelialization in the burn-inducing model.

Test Result 2

1) General symptoms and death of animals

No specific symptoms or deaths were observed by administration of the test animals.

2) Result of burn size measurement

-The results were evaluated by comparing the measured horizontal and vertical lengths and the calculated area values with the untreated group G1 and the negative control group G2.

-As a result of measuring the horizontal and vertical lengths of the burned area, the size increased as shown in FIG.

3) Measurement result of the image width.

The measurement results of the transverse lengths showed the effect that the image area of G3 is smaller than that of G1 and G2 as shown in FIG. 1 and Table 2 below.

Concentration effect (image side size) of mouse image model Changes of the horizontall size (mm) Groups Day 0 Days 2 Days 6 Days 9 Days 13 Days 16 Days 20 G1 14.66
± 0.00 15.00
± 0.25 14.24
± 0.21 13.78
± 0.26 12.59
± 0.46 10.53
± 0.64 7.51
± 0.58 G2 14.66
± 0.00 15.05
± 0.29 14.22
± 0.19 13.33
± 0.15 12.22
± 0.25 9.04
± 0.53 6.51
± 0.50 G3 14.66
± 0.00 15.25
± 0.35 14.64
± 0.17 13.88
± 0.13 12.33
± 0.71 7.70
± 0.67 ^## 4.49
± 0.26 ^{**, ##} G4 14.66
± 0.00 15.24
± 0.24 14.38
± 0.17 13.45
± 0.20 11.57
± 0.57 8.82
± 0.74 4.90
± 0.27 ^{**, ##} G5 14.66
± 0.00 15.50
± 0.30 15.29
± 0.47 13.56
± 0.45 ^# 13.19
± 0.46 11.32
± 0.44 ^## 9.19
± 0.66 ^{**, ##}

Data were expressed as Mean ± S.E.M. The results were statistically analyzed by One-way ANOVA.

*: significantly different from G1, P <0.05;

**: significantly different from G1, P <0.01

# significantly different from G2, P <0.05;

##: significantly different from G2, P <0.01;

G1: Untreated control

G2: Negative control (Distilled water 0.1 ㎖ / head)

G3: Test article 1 (Concentrate 0.1 ml / head)

G4: Test article II (concentrate B 0.1 ml / head)

G5: Positive control (SLIMAZIN 1% CREAM 0.1 ㎖ / head)

That is, as shown in FIGS. 2A to 2E, the horizontal length was statistically significantly smaller than that of the untreated group G1 on the 16th and 20th days (P <0.01). The size was decreased on the 20th day compared to the negative control group (G2) (P <0.01).

Also, as shown in the diagram of FIG. 2D, the concentrate B-administered group (G4) had a smaller wound size than the G1 and G2 on the 20th day (P <0.01 or P <0.05). The positive control group (G5) showed a larger horizontal length than G2 on the 16th and 20th days as illustrated in FIG. 2E (P <0.05 or P <0.01), and the difference was greater on the 20th day compared to G1. (P <0.05)

4) Vertical Length Results

The measurement results of the longitudinal lengths showed the effect that the image area of G3 is smaller than G1 and G2 as shown in FIG. 3 and Table 3 below.

Concentrate effect (image vertical size) of mouse image model Changes of the vertical size (mm) Groups Day 0 Days 2 Days 6 Days 9 Days 13 Days 16 Days 20 G1 14.66
± 0.00 14.83
± 0.34 13.98
± 0.24 13.68
± 0.18 12.53
± 0.39 10.22
± 0.46 7.51
± 0.58 G2 14.66
± 0.00 14.98
± 0.24 14.51
± 0.16 13.76
± 0.15 12.94
± 0.16 9.83
± 0.46 6.51
± 0.50 G3 14.66
± 0.00 15.34
± 0.19 14.57
± 0.18 13.83
± 0.11 12.28
± 0.57 8.26
± 0.49 ^{**, #} 4.49
± 0.26 ^{**, #} G4 14.66
± 0.00 15.02
± 0.29 14.52
± 0.24 13.43
± 0.16 12.03
± 0.37 9.90
± 0.50 4.90
± 0.27 ^{**, ##} G5 14.66
± 0.00 14.77
± 0.13 14.36
± 0.19 13.71
± 0.17 12.99
± 0.25 11.77
± 0.38 ^{*, ##} 9.67
± 0.62 ^{*, ##}

Data were expressed as Mean ± S.E.M. The results were statistically analyzed by One-way ANOVA.

*: significantly different from G1, P <0.05;

**: significantly different from G1, P <0.01

#: significantly different from G2, P <0.05;

##: significantly different from G2, P <0.01;

G1: Untreated control

G2: Negative control (Distilled water 0.1 ㎖ / head)

G3: Test article 1 (Concentrate 0.1 ml / head)

G4: Test article II (concentrate B 0.1 ml / head)

G5: Positive control (SLIMAZIN 1% CREAM 0.1 ㎖ / head)

That is, as shown in the accompanying Figures 4a to 4c the image area of G3 showed a smaller effect on the 16th and 20th day compared to G1 and G2 (P <0.05 or P <0.01),

As shown in the diagram of FIG. 4D, the concentrate B-administered group (G4) had a smaller wound size than the G1 on the 20th day (P <0.05). Positive control group (G5) showed a difference in the healing rate of the wound to a significantly larger value than the G1 and G2 on days 16 and 20 as shown in Figure 4e (P <0.05 or P <0.01).

5) Burnout front result

The measurement results on the front of the image part showed the effect that the image area of G3 is smaller than G1 and G2 as shown in FIG. 5 and Table 4 below.

Concentrate effect (the image front part) of mouse image model Changes of the area size (mm ² ) Groups Day 0 Days 2 Days 6 Days 9 Days 13 Days 16 Days 20 G1 214.92
± 0.00 222.60
± 7.08 198.94
± 3.70 188.49
± 4.47 158.33
± 8.53 108.76
± 9.83 62.19
± 9.04 G2 214.92
± 0.00 225.62
± 7.75 206.15
± 2.28 183.53
± 3.03 158.97
± 4.73 90.82
± 9.28 49.51
± 5.54 G3 214.92
± 0.00 233.75
5.87 213.33
± 4.20 ^* 191.91
± 2.22 154.59
± 13.97 66.20
± 10.80 ^** 25.84
± 3.08 ^{**, #} G4 214.92
± 0.00 229.27
± 7.43 208.77
± 4.02 180.57
± 3.74 140.88
± 10.70 90.23
± 12.29 31.29
± 3.19 ^** G5 214.92
± 0.00 228.97
± 5.14 219.54
± 7.29 ^** 199.96
± 7.67 171.78
± 7.90 134.62
± 8.95 ^## 91.59
± 11.13 ^{**, ##}

Data were expressed as Mean ± S.E.M. The results were statistically analyzed by One-way ANOVA.

*: significantly different from G1, P <0.05;

**: significantly different from G1, P <0.01

# significantly different from G2, P <0.05;

##: significantly different from G2, P <0.01;

G1: Untreated control

G2: Negative control (Distilled water 0.1 ㎖ / head)

G3: Test article 1 (Concentrate 0.1 ml / head)

G4: Test article II (concentrate B 0.1 ml / head)

G5: Positive control (SLIMAZIN 1% CREAM 0.1 ㎖ / head)

In other words, the area values calculated from the horizontal and vertical values measured as shown in the accompanying diagrams of FIGS. 5A to 5C showed greater values than G3 and G1 on day 6, and compared to G1 and G2 on day 16 and 20. The size was small (P <0.05 or P <0.01).

As shown in the diagram of FIG. 5, the concentrate B-administered group (G4) was significantly smaller than the G1 on the 20th day (P <0.01). The positive control group (G5) showed a difference greater than G1 and G2 on days 6, 16, and 20 as shown in FIG. 5 (P <0.01)

6) Histopathology test result

-Evaluation of skin burn cure is shown in Fig. 7 epithelialization, inflammation, neovascularization, activation of fibroblasts, deposition of collagen and wound contraction. (wound contraction) and the detailed effects are shown in Table 5.

Histopathological scoring in a burn mouse model Histopathological criteria for the evaluation of cutaneous burn Groups Epithelia
lization Inflammat
ion Neovascul
arization Activation of
fivroblasts Collagen
deposition Wound
contraction G1 0.2 ± 0.2 2.8 ± 0.1 2.5 ± 0.2 2.7 ± 0.2 2.1 ± 0.1 0.9 ± 0.4 G2 1.6 ± 0.2 2.9 ± 0.1 2.7 ± 0.2 3.1 ± 0.1 2.1 ± 0.1 2.2 ± 0.4 ^** G3 3.1
± 0.2 ^{**, ##} 2.4 ± 0.2 1.9 ± 0.2 2.2 ± 0.2 ^# 2.4 ± 0.2 1.5 ± 0.2 G4 3.0
± 0.3 ^{**, ##} 2.6 ± 0.2 2.0 ± 0.3 2.7 ± 0.2 2.4 ± 0.2 2.3 ± 0.2 ^** G5 1.3 ± 0.2 1.7
± 0.2 ^{**, ##} 2.0 ± 0.1 ^# 2.1
± 0.1 ^{*, ##} 2.0 ± 0.1 1.8 ± 0.2

* 0: No microscopic findings; 1: Minimal; 2: Mild; 3: Moderate 4: Marked

Data were expressed as Mean ± S.E.M. The results were statistically analyzed by One-way ANOVA.

*: significantly different from G1, P <0.05;

**: significantly different from G1, P <0.01

# significantly different from G2, P <0.05;

##: significantly different from G2, P <0.01;

G1: Untreated control

G2: Negative control (Distilled water 0.1 ㎖ / head)

G3: Test article 1 (Concentrate 0.1 ml / head)

G4: Test article II (concentrate B 0.1 ml / head)

G5: Positive control (SLIMAZIN 1% CREAM 0.1 ㎖ / head)

That is, the degree of formation of the observed findings was divided into five stages, ranging from 0 points without findings to 4 points showing the most significant findings.

-As a result of comparing the average value of histopathological criteria for the evaluation of burn healing in each group, as shown in FIGS. 8A to 8D, the degree of epithelialization was higher in the G3 and G4 test group compared to G1 and G2. Higher (P <0.01)

Inflammation was not different between the untreated group (G1) and the negative control group (G2), and the degree of neovascularization was not found in the cisar concentrate (G3) and concentrate stock solution B (G4) groups. A slight tendency appeared.

In addition, activation of fibroblasts was lower than that of the negative control group (G2) (P <0.05), and collagen deposition did not show any difference.

Wound contraction showed an increase in negative control group (G2) and concentrate undiluted solution B (G4) compared to untreated bacteria (G1) (P <0.01).

As shown in FIG. 8E, the positive control group G5 did not show any difference from the untreated bacteria (G1) in the degree of epithelialization, but the inflammation was significantly reduced compared to the untreated bacteria (G1) and the negative control group (G2). Laziness (P <0.01).

Neovascularization and Activation of Fibroblasts were found to be lower (P <0.05 or P, 0.01) than collagen (G1) and negative control (G2) and collagen deposition. ), There was no difference.

On the other hand, the positive control group (G5) showed a slight increase in wound contraction compared to the untreated group (G1).

Test Result 3

In the present invention, a Sparague-Dawley-based rat was used to verify the healing effect of the test substance cisar concentrate and concentrate undiluted solution B on the burn model induced by surgical treatment.

The healing process for skin damage proceeds in the order of three stages: the Inflammatory phase, the Proliferative phase, and the Remodeling phase (Kumar V, Abbas Ak, Fausto N. (2005)). Robbins and Cotran pathologic basis of disease.Elsevier Inc .; Jones TC, Hunt RD, King NW. (1997): Veterinary pathology.Willians & Wilkins).

 In general, the inflammatory phase is a process in which fibrin is formed by platelet activity, and a crust is formed, and neutrophils are initially infiltrated with hemostasis, and macrophage infiltration is shown as time passes. (Cohen IK, Diegelmann RF, Lindblad WJ. (1992): Wound healing: Biochemical & clinical aspects.WB Saunders, Harcourt Brace Jovanovich Inc.)

 The middle age is the beginning of granulation tissue formation. As inflammatory findings decrease, fibroblast proliferation, angiogenesis and collagen deposition are observed in the damaged area. As the process of the Chinese food progresses, the amount of collagen in the damaged area gradually increases, and epidermalization of the epidermis proceeds. The remodeling phase gradually repairs the epidermal layer to normal thickness and damage as collagen deposition is properly controlled.

In some cases, excessive collagen deposition, epithelial hyperplasia, or contraction of the wound may result in scars that do not recover and scars remain. (Cohen IK, Diegelmann RF, Lindblad WJ. (1992): Wound healing: Biochemical & clinical aspect.WB Saunders, Harcourt Brace Jovanovich Inc.)

Therefore, the present invention was applied to the burn site for 3 weeks after causing the burn in the rat, while measuring the size of the burn site of each animal during the healing progress, and histopathological examination after the end of the treatment, the results are not treated group (G1) and negative control (2).

In the present invention, histopathological examination is generally performed 21 days after the induction of burns, and histopathologically related to the remodeling stage. Persistence and angiogenesis, fibroblast activation and collagen accumulation were assessed separately. The wound site contraction was also compared.

As a result of measuring the size of the burn site, the congestion group according to the present invention showed significantly smaller values than the untreated group and the negative control group on the 16th and 20th days after the induction. Histopathologically, the progression of epithelialization was significantly accelerated compared to the untreated group.

This showed better wound healing compared to the positive control group. In other words, there was no difference in inflammation reduction, neovascularization, collagen deposition, and wound contraction with no treatment group, and there was a decrease in fibroblast activation.

Outlet rain stock solution B, another test substance of the present invention, had an effect of reducing the size of the wound on the 20th day, and histopathological examination showed a high degree of epithelialization and wound contraction.

It may be considered that the large wound contraction in wound healing may indicate scars, but it is difficult to have great toxicological meaning because the deviation of the untreated group is large and the negative control group is increased.

In other words, the test substance had no significant effect on the inhibition of inflammation or the proliferation and recovery of the dermal layer at the time of necropsy.

In contrast, the positive control group had no effect on epithelialization, but the inflammatory response was significantly suppressed. Silmargin 1% cream, used as a positive control, is a silver suifadiazine and is the most prescribed drug to prevent secondary infection of the burned area. However, due to this anti-inflammatory action is known to cause side effects that delay the healing of wounds.

Taken together, it was confirmed that the concentrate and the concentrate undiluted solution B had an effect of reducing the size of the window and promoting the progression of epithelialization in the burn-inducing model.

The invention being thus described, it will be obvious that the same way may be varied in many ways. Such modifications are intended to be within the spirit and scope of the invention as defined by the appended claims.

Claims (4)

A composition for treating burns, comprising citrate, zinc and L-arginine as active ingredients in a therapeutically effective amount together with a pharmaceutically acceptable carrier and diluent.
The method of claim 1,
The composition for treating burns, comprising 0.02 to 20% by weight of citric acid, 0.0001 to 1.0% by weight of zinc and 0.025 to 25% by weight of L-arginine as the active ingredient.
The method according to claim 1 or 2,
A composition for treating burns, comprising a pharmaceutically acceptable ointment in addition to the composition.
The method of claim 3, wherein
The ointment is burn fat composition, characterized in that selected from petrolatum, paraffin, vegetable oil, lard, wax and sweet ointment.

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