Topical skin composition effective in promoting wound healing and / or scar repair - Patent Application 20070122997

Abstract

The present invention relates to a topical skin composition effective in promoting wound healing and / or scar repair, comprising (A) concentrated birch sap and (B) one or more active substances selected from burnet root extract, coptis root extract, lilac extract, allantoin, red sage extract, astragalus extract, pearl powder, and asiaticoside, wherein the concentrated birch sap is concentrated by about 1.2 to 6 times, preferably about 1.5 to 5 times, and more preferably about 1.5 to 3 times.

Description

[Technical Field]

[0001] Technical Field The present invention relates to a topical skin composition effective in promoting wound healing and / or scar repair, comprising (A) concentrated birch sap and (B) one or more active substances selected from Sanguisorba officinalis root extract, Coptis japonica root extract, Lithospermum erythrorhizon extract, allantoin, Salvia miltiorrhiza extract, Astragalus extract, pearl powder, and asiaticoside, wherein the concentrated birch sap has a concentration of about 1.2 to 6 times, preferably about 1.5 to 5 times, and more preferably about 1.5 to 3 times. [Background technology]

[0002] Background technology Skin burns, scalding, trauma, and other injuries are common skin diseases that threaten human health and life safety. The resulting skin wounds can damage the integrity of the skin epithelium and impair the structure and function of the underlying normal tissues. The wound healing process often results in pain in the affected area, delayed healing, infection and inflammation, and high body fever, causing distress to the patient. Promoting wound healing is a complex and expensive process. Research into drugs that promote wound healing is a developing field in modern biomedical science. Currently, a range of drugs containing various medicinal ingredients with wound-healing and antibacterial potential, including ointments, creams, gels, and sprays, are widely used to promote wound healing. However, problems such as low healing rates and high recurrence rates still exist.

[0003] Scars are caused by physical, biological, chemical, and other factors. Particularly after burns, scalding, and trauma, human skin and soft tissue lesions often extend beyond the original skin injury, forming persistent nodular, threadlike, or flaky masses of tissue that are elevated above the skin surface, have a hard texture, and are hyperemic. Years of scar hyperplasia often plague patients. The subsequent period of atrophy leaves patients completely disfigured and disabled, resulting in significant physical and mental disabilities. Currently, the main methods for improving or treating scars include gene therapy, intrascar injections, oral anti-scarring drugs, cryotherapy, pressure therapy, laser treatment, silicone gel membrane products, and radiation therapy. The therapeutic efficacy of various treatment methods remains uncertain. However, all of these methods suffer from the drawbacks of slow effectiveness, long treatment cycles, inherently low mobility, and severe side effects. Summary of the Invention [Problem to be solved by the invention]

[0004] Thus, there remains a need for improved methods and products in the field of promoting wound healing and / or scar repair.

[0005] Wound healing is a complex and dynamic process. The cellular structure and tissue layers of damaged tissue are restored to as normal as possible. This is a highly coordinated and regulated complex biological process involving various cells, extracellular matrix, and cytokines. Wound healing can be roughly divided into four overlapping stages: hemostasis, inflammation, proliferation, and remodeling. Hemostasis and inflammation are two distinct features of the inflammatory phase, while the proliferative phase involves epithelialization, angiogenesis, and collagen deposition. Scarring may occur during tissue wound contraction in the maturation stage. Reducing inflammation and promoting fibroblast proliferation and angiogenesis will contribute to rapid wound healing. The pathological basis of hypertrophic scarring is excessive proliferation of fibroblast-based cellular components and excessive deposition of collagen-based extracellular matrix. Controlling the inflammatory response and reducing excessive deposition of extracellular collagen can reduce and repair scarring.

[0006] Birch sap is a sap derived from the Betulaceae family that is rich in active ingredients such as polysaccharides, amino acids, vitamins, biotin, cytokinins, minerals, and trace elements required by the human body. There are records in Europe of the use of birch sap to treat burns and scalding wounds. As a natural ingredient, birch sap has great potential in promoting wound healing and repairing scars. [Means for solving the problem]

[0007] Therefore, the present inventors have completed the present invention by testing the effect of birch sap in promoting wound healing and / or scar repair. [Brief explanation of the drawings]

[0008] [Figure 1] 1 is a graph showing the cell migration rate obtained in Example 2. [Figure 2] 1 is a graph showing the degree of wound healing obtained in Example 3. DETAILED DESCRIPTION OF THE INVENTION

[0009] Summary of the Invention In one aspect, the present invention relates to the use of concentrated birch sap in a topical skin composition having the efficacy of promoting wound healing and / or scar repair, wherein the concentrated birch sap has a concentration of about 1.2 to 6 times, preferably about 1.5 to 5 times, more preferably about 1.5 to 3 times.

[0010] In another aspect, the present invention relates to a topical skin composition effective in promoting wound healing and / or scar repair, comprising (A) concentrated birch sap and (B) one or more active substances selected from burnet root extract, coptis root extract, lilac extract, allantoin, red sage extract, astragalus extract, pearl powder, and asiaticoside, wherein the concentrated birch sap has a concentration of about 1.2 to 6 times, preferably about 1.5 to 5 times, and more preferably about 1.5 to 3 times.

[0011] The birch sap involved in the present invention is obtained from the genus Betulaceae, including four varieties: Betula alba, Betula pubescens, Betula Pendula, and Betula platyphylla. Birch sap is a clear, colorless, and nutritious birch juice containing no precipitants or impurities, obtained by artificially drilling and collecting from the base of birch tree trunks from the time when snow begins to melt until the tree leaves. Birch sap is commercially available and can be used as is (referred to as undiluted birch sap in this case) and can be purchased, for example, from Daxinganling Chaoyue Wild Berry Development Co., Ltd.

[0012] The concentrated birch sap used in the present invention can be obtained by concentrating the commercially available birch sap concentrate product described above. Concentration methods are known in the art, such as heat concentration, low-temperature and vacuum concentration, and membrane concentration. In the present invention, concentration is preferably carried out by low-temperature freeze concentration or membrane concentration. For example, commercially available birch sap concentrate is introduced into a low-temperature drying device, cooled to about -40 to -70°C, and evacuated to about 0.1 to 30 Pa for the low-temperature and vacuum concentration method to obtain concentrated birch sap with different concentrations.

[0013] The content of component (A), concentrated birch sap, in the topical skin composition is about 10 to 98 wt %, preferably 20 to 98 wt %, and more preferably about 30 to 97 wt %, based on the total weight of the topical skin composition.

[0014] Furthermore, the inventors have unexpectedly discovered that, compared with the use of concentrated birch sap, burnet root extract, Coptis Rhizome root extract, lizard extract, allantoin, Red Sage extract, Astragalus extract, pearl powder, and asiaticoside alone, the use of concentrated birch sap in combination with one or more selected from burnet root extract, Coptis Rhizome root extract, lizard extract, allantoin, Red Sage extract, Astragalus extract, pearl powder, and asiaticoside exhibits significantly greater effectiveness in promoting wound healing and / or scar repair than the additive effects of these, as shown by better fibroblast migration, promotion of wound healing rate in a skin defect model rat, inhibition of inflammation-related TNF-α and IL-6 protein content, reduction in scar hyperplasia index and scar tissue collagen deposition and the degree of physical scarring in rabbits, indicating the existence of a synergistic effect between concentrated birch sap and the above active substances.

[0015] Therefore, in another aspect, the present invention relates to the use of (A) concentrated birch sap in combination with (B) one or more active substances selected from Sanguisorba officinalis root extract, Coptis japonica root extract, Lithospermum parkii extract, allantoin, Salvia miltiorrhiza extract, Astragalus ulmoides extract, pearl powder, and asiaticoside in a topical skin composition effective in promoting wound healing and / or scar repair, wherein the concentrated birch sap has a concentration of about 1.2 to 6 times, preferably about 1.5 to 5 times, and more preferably about 1.5 to 3 times.

[0016] In another aspect, the present invention relates to a topical skin composition effective in promoting wound healing and / or scar repair, comprising (A) concentrated birch sap and (B) one or more active substances selected from burnet root extract, coptis root extract, lilac extract, allantoin, red sage extract, astragalus extract, pearl powder, and asiaticoside, wherein the concentrated birch sap has a concentration of about 1.2 to 6 times, preferably about 1.5 to 5 times, and more preferably about 1.5 to 3 times.

[0017] Component (B) Sanguisorba officinalis root extract, Coptis japonica root extract, Lithospermum officinalis extract, allantoin, Salvia miltiorrhiza extract, Astragalus extract, pearl powder, and asiaticoside are known in the art, are all commercially available, and are used as is in the present invention.

[0018] The total content of component (B) in the topical skin composition is approximately 0.0005 to 30%, preferably approximately 0.001 to 10%, more preferably approximately 0.1 to 5%, and most preferably approximately 0.5 to 3%, based on the total weight of the topical skin composition.

[0019] The topical skin compositions include pharmaceutical compositions and cosmetic skin care compositions, where the pharmaceutical compositions exhibit significant wound healing and scar repair efficacy, and the cosmetic skin care compositions exhibit significant scar repair efficacy.

[0020] The topical skin composition does not contain any additionally added water, but does not exclude the water inherently contained in each of the components.

[0021] In one preferred embodiment, the topical skin composition is free of chelating agents, such as EDTA salts, sodium phosphate, sodium metaphosphate, and gluconic acid.

[0022] In addition to the above-mentioned components (A) and (B), the topical skin composition may optionally contain (C) a component commonly used in compositions for wound healing and scar repair. Examples of component (C) include, but are not limited to, a vehicle, an active ingredient, an excipient, etc. These components are known in the art, and their types and amounts can be selected by those skilled in the art as needed. For example, the content of component (C) is typically about 0 to 70 wt % based on the total weight of the topical skin composition.

[0023] Vehicles include, for example, diluents, dispersants, or carriers. Examples include, but are not limited to, ethanol, dipropylene glycol, butanediol, etc. The content of the vehicle in topical skin compositions is known in the art, and is, for example, typically about 0.5 to 20% of the total weight of component (C).

[0024] Active ingredients include antibacterial agents, anti-inflammatory agents, astringents, moisturizers, and the like. Antibacterial agents include, but are not limited to, one or more of ursolic acid, oldenlandia diffusa flavonoids, honeysuckle flower extract, tea tree essential oil, chitin, cinnamon branch extract, coral ginger volatile oil, clove extract, mushroom extract, aloe extract, mugwort leaf extract, 1-pentadecanol and its derivatives, sedalene, caryophyllene, and phyllene. The content of the antibacterial agent in a topical skin composition is known in the art, and is typically about 0.01 to 30% of the total weight of component (C).

[0025] Anti-inflammatory agents include, but are not limited to, one or more of safflower yellow, dipotassium glycyrrhizinate, cattail pollen extract, arrowroot extract, cucumber extract, garlic extract, burdock extract, resveratrol, Sophora flavescens extract, magnolia bark extract, etc. The content of the anti-inflammatory agent in a topical skin composition is known in the art, and is typically about 0.01 to 50% of the total weight of component (C).

[0026] Astringents include, but are not limited to, one or more of green tea polyphenols, witch hazel extract, vitamin A, menthol lactate, seaweed extract, blue jelly, sulfur, rehmannia root, angelica, lactic acid, tartaric acid, succinic acid, citric acid, etc. The amount of astringent in a topical skin composition is known in the art, and is typically about 0.01 to 30% of the total weight of component (C).

[0027] Examples of humectants include, but are not limited to, glycerin, diglycerin, butylene glycol, propylene glycol, 1,3-propanediol, dipropylene glycol, 1,2-pentanediol, polyethylene glycol-8, polyethylene glycol-32, methyl gluceth-10, methyl gluceth-20, PEG / PPG-17 / 6 copolymer, glycereth-7, glycereth-26, glyceryl glucoside, PPG-10 methyl glucose ether, PPG-20 methyl glucose ether Examples of humectants include one or more of: glycerol, PEG / PPG / polybutylene glycol-8 / 5 / 3 glycerol, sucrose, trehalose, rhamnose, mannose, raffinose, betaine, erythritol, xylitol, urea, glycereth-5 lactate, sodium hyaluronate, hydrolyzed sodium hyaluronate, acetylated sodium hyaluronate, sodium polyglutamate, hydrolyzed sclerotium gum, pullulan, tremellam, and tamarind seed polysaccharide. The amount of humectant in a topical skin composition is known in the art; for example, it is typically about 0.01 to 30% of the total weight of component (C).

[0028] Excipients are known in the art and include, for example, surfactants, emulsifiers, thickeners, preservatives, flavoring agents, and the like.

[0029] Examples of surfactants include, but are not limited to, cocamidopropyl betaine, sodium laureth sulfate, PEG-150 distearate, ethylene glycol distearate, ammonium laureth sulfate, sodium lauryl ether sulfate, palmamidopropyl betaine, cocamide diethanolamine, lauryl ether sulfate, ammonium lauryl sulfate (K12A), sodium fatty alcohol polyvinyl ether sulfate (AES), ammonium fatty alcohol polyvinyl ether sulfate (AESA), ammonium laureth sulfate, ammonium lauryl sulfate, sodium lauryl sulfate, ammonium lauryl polyether sulfate, ammonium dodecyl sulfate, cocomono Examples of surfactants include one or more of ethanolamides, N-fatty acyl amino acid salts, lauramidopropyl betaine, sodium cocoamphoacetate, sodium lauroamphoacetate, sodium laureth sulfate, sodium fatty alcohol polyoxyethylene ether sulfate, sodium lauryl sulfate, alkyl polyglucosides, lauryl dimethyl betaine, disodium imidazolinylamphodiacetate, sodium cocoamphoacetate, polyquaternium-7, polyquaternium-10, polyquaternium-37, hydroxypropyl guar gum hydroxypropyltrimethylammonium chloride, hydroxypropyltrimethylammonium chloride guar gum, linden seed extract, etc. The content of surfactants in topical skin compositions is known in the art and is typically about 0.1 to 50% of the total weight of component (C).

[0030] Examples of emulsifiers include, but are not limited to, cetearyl olivate, sorbitan olivate, polysorbate-60, polysorbate-80, methyl glucose sesquistearate, PEG-20 methyl glucose sesquistearate, PEG-40 hydrogenated castor oil, PPG-26-buteth-26, PEG-4 polyglyceryl-2 stearate, PEG-60 hydrogenated castor oil, steareth-2, steareth-21, PPG-13-decyltetradeceth-24, cetearyl glucoside, PEG-100 stearate, glyceryl stearate, glyceryl stearate SE, Examples of the emulsifier include one or more of coco-glucoside, ceteareth-25, PEG-40 stearate, polyglyceryl-3 methylglucose distearate, glyceryl stearate citrate, polyglyceryl-10 stearate, polyglyceryl-10 myristate, polyglyceryl-10 dioleate, polyglyceryl-10 laurate, polyglyceryl-10 isostearate, polyglyceryl-10 oleate, polyglyceryl-10 diisostearate, polyglyceryl-6 laurate, polyglyceryl-6 myristate, sucrose stearate, and sucrose polystearate. The content of the emulsifier in a topical skin composition is known in the art, and is typically 0.05 to 30% of the total weight of component (C).

[0031] Examples of thickeners include, but are not limited to, one or more of high molecular weight polymers such as carbomer, acrylates and their derivatives, xanthan gum, gum arabic, polyethylene glycol-14M, polyethylene glycol-90M, succinyl polysaccharides, hydroxyethyl cellulose, hydroxypropyl cellulose, and hydroxypropyl methylcellulose. The content of thickeners in topical skin compositions is known in the art, and is typically 0.1 to 30% by weight of the total weight of component (C).

[0032] Examples of preservatives include, but are not limited to, one or more of methyl hydroxybenzoate, ethyl hydroxybenzoate, propyl hydroxybenzoate, phenoxyethanol, benzyl alcohol, phenylethanol, bis(hydroxymethyl)imidazolidinyl urea, potassium sorbate, sodium benzoate, chlorophenesin, and sodium dehydroacetate. The content of preservatives in topical skin compositions is known in the art, and is typically 0.01 to 30% of the total weight of component (C).

[0033] The topical skin composition of the present invention can be prepared by any suitable method known in the art. For example, it can be prepared using devices commonly used in the cosmetics field, such as a dissolving tank, emulsifying pot, disperser, transfer pump, etc. During preparation, the water-soluble material is placed in the aqueous phase dissolving kettle, and the oil-soluble material is placed in the oil phase dissolving kettle. The two kettles are each heated to approximately 80°C. For easily coagulating ingredients, a disperser can be used to pre-disperse them. Once dissolution is complete, the oil and aqueous phases are transferred to the emulsifying pot and homogenized and emulsified for approximately 5 to 15 minutes. Once emulsification is complete, the bulk is cooled to room temperature, and fragrances, preservatives, etc. are optionally added, and the pH of the product is adjusted as needed. The above preparation method can be modified or adjusted depending on the requirements of the product's formulation.

[0034] Pharmaceutical or cosmetic compositions can be prepared in various dosage forms, such as liquids, lotions, ointments, creams, or gels, as needed, and cosmetic compositions can be in various forms, such as lotions, sprays, emulsions, extracts, milks, and creams. [Example]

[0035] Example The present invention will be described in more detail below with reference to examples.However, it should be understood that these examples and comparative examples are only used to specifically illustrate the present invention, and should not be understood to limit the scope of the appended claims of the present invention in any way.

[0036] Example 1: Determination of cell viability The cells used in the test were human-derived fibroblasts obtained from Guangdong Boxi Biotechnology Co., Ltd. through primary and subculture.

[0037] 1.1 Reagents Cell culture medium (Gibco), neonatal bovine serum (Sijiqing), PBS (Boster), MTT (Sigma), DMSO (Sigma).

[0038] 1.2 Main equipment CO2 incubator (Thermo), clean bench (AIRTECH), inverted microscope (Olympus), micro-oscillator (QLBER), microplate reader (BioTek), incubator (Taisite).

[0039] 1.3 Test Sample Fresh birch (Betula alba) sap stock solution purchased from Daxinganling Chaoyue Wild Berry Development Co., Ltd. was introduced into a low-temperature drying device, cooled to -65°C, evacuated to 0.1 Pa, and concentrated to 1.2, 2, 3, and 5 times the concentration.

[0040] 1.4 Determination of cell viability (1) Cell inoculation: Cells were inoculated into 96-well culture plates at a density of 1E4 cells / well and incubated overnight in an incubator (37°C, 5% CO2, 95% relative humidity); Experimental grouping: A blank control group, a positive control group, and a sample group were established. When the cell seeding rate of the 96-well plate reached 40-60%, the prepared culture medium containing the test sample was administered to the groups at a volume of 200 μL per well, with three replicate wells per group. In the positive control group, 200 μL of culture medium containing dermatan sulfate (5 μM) was added to each well. In the sample group, 200 μL of culture medium containing the corresponding concentration of the test substance was added to each well. The zero adjustment well was not inoculated with cells, but instead contained 200 μL of cell culture medium alone. After administration, the 96-well plate was placed in an incubator (37°C, 5% CO2, 95% relative humidity).

[0041] 1.5 Decision After incubating the cells for 24 hours, the supernatant was discarded, and MTT working solution (0.5 mg / mL, ready to use) was added and incubated in the dark for 4 hours at 37°C. At the end of the incubation, the supernatant was discarded, 150 μL of DMSO was added to each well, and the OD value was read at 490 nm.

[0042] The results of the test are shown in the table below.

[0043] [Table 1]

[0044] The above experimental results showed that concentrated birch sap with a concentration of 1.2 to 5 times compared with aqueous birch sap and undiluted birch sap significantly improved the proliferation ability of fibroblasts, and that the viability of fibroblasts was highly promoted, especially when the concentration was 2 to 3 times.

[0045] Example 2: Cell migration experiments 2.1 Cell sources The cells used in the test were human-derived fibroblasts obtained from Guangdong Boxi Biotechnology Co., Ltd. through primary and subculture.

[0046] 2.2 Reagents Cell culture medium (Gibco), newborn bovine serum (Sijiqing), PBS (Boster), 60 mm culture dishes (Coring), fibronectin (Sigma).

[0047] 2.3 Main equipment Phase contrast microscope (Olympus) 2.4 Experimental samples Fresh birch (Betula alba) sap stock solution purchased from Daxinganling Chaoyue Wild Berry Development Co., Ltd. was introduced into a low-temperature drying device, cooled to −65°C, evacuated to 0.1 Pa, and concentrated three times.

[0048] When loading samples, the single ingredient load was the full amount of ingredient, and the blended ingredients were half birch sap ingredient plus half 0.1% of other active ingredients. The samples tested included: 3x concentrated birch sap, burnet root extract, coptis root extract, 3x concentrated birch sap + burnet root extract, and 3x concentrated birch sap + coptis root extract.

[0049] 2.3 Experimental steps (1) 60 mm culture dishes were coated with fibronectin overnight.

[0050] (2) The coated culture dish was washed three times with PBS to remove excess fibronectin.

[0051] (3) Logarithmic phase cells were digested and counted, and then 2E6 cells were seeded onto the coated culture dish for 48 hours. After the cells covered the bottom, a straight scratch was quickly made on the bottom of the culture dish using a 200 μL pipette tip. Excess medium was aspirated, and excess cells were washed away with PBS. Cell culture medium was added again for incubation, and the test samples with different concentrations were added for stimulation.

[0052] (4) The cells were returned to a 37°C incubator for culture, and the distance of cell movement was recorded and measured every 12 hours using a phase-contrast microscope.

[0053] (5) Data processing: total migration distance of cells at each time point = distance length at time 0 - distance length at each time point.

[0054] The experimental results are shown in Figure 1. The results showed that the combination of 3x concentrated birch sap with Sanguisorba officinalis root extract or Coptis japonica root extract significantly promoted fibroblast migration compared to the use of 3x concentrated birch sap, Sanguisorba officinalis root extract, and Coptis japonica root extract alone.

[0055] Example 3: Rat full-thickness skin defect wound model 3.1 Experimental animals Six- to seven-week-old male Sprague Dawley (SD) rats, half male and half female, were purchased and fed for one week until the rats adapted to the experimental environment.

[0056] 3.2 Experimental samples Fresh birch (Betula alba) sap stock solution purchased from Daxinganling Chaoyue Wild Berry Development Co., Ltd. was introduced into a low-temperature drying device, cooled to −65°C, evacuated to 0.1 Pa, and concentrated to about 1.5 times.

[0057] When loading samples, the single ingredient load was the full amount of ingredient, and the blended ingredients were half birch sap ingredient plus half 0.1% of other active ingredients. Samples tested included: 1.5x concentrated birch sap, purple extract, allantoin, 1.5x concentrated birch sap + purple extract, and 1.5x concentrated birch sap + allantoin.

[0058] 3.3 Modeling Method (1) SD rats were randomly divided into a blank control group and an experimental group, each with six rats. The rats were anesthetized with isoflurane inhalation and the experiment was performed when they were completely relaxed, their muscles were relaxed, and their corneal reflex and pain response were abolished.

[0059] (2) The hair on the back of the rat was pierced with a shaver to expose the back, and a wound was marked on the back using a marker.

[0060] (3) Using the pre-made markings and a surgical blade and forceps, cut the area approximately 500 mm 2 An excision wound was made on the back of the rat with a surface area of ​​1000 mm and a depth of 2 mm.

[0061] (4) After disinfection with alcohol, the control group was treated with PBS or Ringer's solution, the wound surface was covered with 3M transparent film, and the test group was sprayed with different concentrations of test samples. The wounds were treated with medication once a day.

[0062] (5) Wound healing was recorded by camera, and the incision wound healing model was counted as day 0, followed by photography and recording on days 7, 14, and 21. The wound healing rate was calculated as follows: wound healing rate = (original wound area - unhealed wound area at each time point) / original wound area.

[0063] (6) After photography and recording on the 20th day, the rats were sacrificed, and the wound area and 2 mm of normal tissue surrounding the wound were collected. PBS was added at a ratio of 1:9 and homogenized using an electric grinder at room temperature to prepare a homogenate. The homogenate was centrifuged at 10,000 rpm at 4°C for 10 minutes, and the supernatant was transferred to a 1.5 mL centrifuge tube and stored at -70°C for later use. The concentrations of TNF-α and IL-6 in the wound tissue were detected by ELISA within 30 days.

[0064] The experimental results were as follows: 1. Extent of wound healing: The results are shown in Figure 2.

[0065] 2. TNF-α, IL-6 concentration

[0066] [Table 2]

[0067] The results of wound healing and TNF-α and IL-6 contents showed that the combination of 1.5x concentrated birch sap with purple extract or allantoin at the same total amount was more effective in promoting tissue regeneration, reducing inflammation, and accelerating wound healing than the use of 1.5x concentrated birch sap, purple extract, or allantoin alone.

[0068] Example 4: Scar model 4.1 Experimental animals Common white rabbits, half male and half female, weighing 2.5–3.0 kg, were housed in accordance with the regulations of the People's Republic of China for the care of laboratory animals. Food and drink were allowed ad libitum every day. The temperature of the feeding room was maintained at 25°C ± 1°C, humidity was maintained at 60–70%, and the daily light / dark cycle was 12 / 12 h per day. The rabbits were fed for 1 week.

[0069] 4.2 Main equipment Digital thickness gauge, multi-function microplate reader, high-throughput tissue grinding equipment, slicer, tissue dehydrator, tissue embedding machine, microscope.

[0070] 4.3 Experimental samples Fresh birch (Betula alba) sap stock solution purchased from Daxinganling Chaoyue Wild Berry Development Co., Ltd. was introduced into a low-temperature drying device, cooled to −65°C, evacuated to 0.1 Pa, and concentrated to about 1.5 times.

[0071] When loading samples, the load of a single ingredient was the total amount of the ingredient, and the blended ingredients were half birch sap ingredient plus half 0.1% of other active ingredients. The samples tested included: 1.5x concentrated birch sap, Red Sage extract, Astragalus extract, 1.5x concentrated birch sap + Red Sage extract, and 1.5x concentrated birch sap + Astragalus extract.

[0072] 4.4 Preparation of rabbit ear hypertrophic scar model The animals were adaptively housed for one week and anesthetized with 1.0 mL / kg (30 mg / kg) of 30 g / L pentobarbital sodium via the ear vein. The skin on the anterior surface of the rabbit ear was disinfected with iodophor and ethanol. In the medial section of the anterior surface of the ear, avoiding visible blood vessels along the longitudinal axis, 1 cm x 1 cm wounds were gently perforated with a corneal ring at intervals of more than 1 cm. Full-thickness skin was removed from the rabbit ear, and the perichondrium was completely scraped off using a spatula. Bleeding was stopped by applying pressure with four sterile cotton balls in each ear, and the wounds were exposed after surgery. After surgery, penicillin was injected intramuscularly on two consecutive days at a dose of 40,000 units / kg. The rabbits were fed normally for one week until the wounds had healed.

[0073] 4.5 Grouping and Dosing After 21 days of the surgical model, scabs and scar formation occurred on the wound surface. The model animals were divided into groups with 6 rabbits in each group: the model group was kept normally without any treatment, and the treatment groups included the model group (PBS group) and the treatment groups with different samples, which were administered by topical application of 0.1 ml to each scar once a day.

[0074] 4.6 Test Indicators (1) Measurement of hypertrophic scar index After 30 days of administration, samples were taken from the center of the scar and the surrounding tissue in the rabbit ear, and the sections were stained with hematoxylin and eosin, then photographed under a microscope, and the scar hyperplasia index was measured and calculated: scar hyperplasia index = the vertical distance from the highest point of the scar to the surface of the ear cartilage tissue / thickness of the surrounding normal skin.

[0075] (2) Scar tissue collagen (hydroxyproline HPr) content After 30 days of administration, the scar tissue of the rabbit ear was removed, and 10% tissue homogenate was prepared conventionally, and the content of hydroxyproline was determined using a kit to investigate the degree of collagen deposition and fibrosis. In addition, the same size of skin and subcutaneous tissue was cut from the other side of the model group (the ear without modeling), and tissue homogenate was prepared by the same method, and the content of hydroxyproline was determined as the normal control group.

[0076] The experimental results are shown as follows:

[0077] [Table 3]

[0078] The results of scar hyperplasia index and hydroxyproline content showed that the combination of 1.5x concentrated birch sap with Red Sage extract or Astragalus extract was more significantly effective in inhibiting scar hyperplasia and reducing scar collagen deposition than the use of 1.5x concentrated birch sap, Red Sage extract, or Astragalus extract alone.

[0079] Example 5: Observation of the therapeutic effect on the human body Scar scores were assessed using the Vancouver Scar Scale (VSS) in four aspects: color, thickness, vascularity, and softness. The total score was 15 points, with higher scores indicating more severe scarring. SPSS statistical software was used for statistical analysis. Measurement data were expressed as x±s. Paired t-tests were used for pre- and post-treatment comparisons within groups, and group t-tests were used for comparisons between groups. Chi-squared tests were used to compare count data between groups. P<0.05 was considered statistically different.

[0080] [Table 4]

[0081] The results of human trials show that the combination of double concentrated birch sap and pearl powder or asiaticoside at the same total dose shows more significant scar-improving effectiveness compared to the use of double concentrated birch sap, pearl powder or asiaticoside alone.

[0082] Example 6: Scar ointment for topical application to the skin The formulation of the scar ointment for topical application to the skin is given in the table below:

[0083] [Table 5]

[0084] The preparation method for the above scar ointment for topical application to the skin was as follows: (1) In a clean flask, add 15g of Danshen extract and 150g of double-concentrated birch sap and stir to dissolve; (2) 125 g of hydroxypropyl-β-cyclodextrin was added to the flask from step (1) and stirred to dissolve evenly; (3) The solution obtained in step (2) was kept in a refrigerator for 90 minutes, suction filtered, the filtrate was poured into a culture dish, covered with perforated plastic wrap, frozen in a refrigerator at -80°C for 24 hours, dried in a vacuum freeze dryer, and when the mixture became a white fluffy mass, it was taken out for use; (4) 160g of stearic acid, 100g of glyceryl monostearate, 35g of cetyl alcohol, 35g of stearyl alcohol, 80g of white petrolatum and 40g of lanolin were mixed, heated to melt, and kept at a temperature of 80°C; (5) 75 g of glycerol, 12 g of triethanolamine, 7 g of Tween-80, 2 g of methylparaben, 2 g of ethylparaben, and 12 g of borneol were weighed, and 150 g of double-concentrated birch sap was added thereto, and the mixture was heated to dissolve. The mixture was then slowly added to the oil phase of step (4) with stirring at 80°C, and the mixture was stirred to emulsify the base. The product of step (3) was then added, and the mixture was stirred until it reached room temperature and uniform.

[0085] Forty subjects were treated with the scar ointment of the above formula for three months and then subjectively evaluated. The results showed that 30 subjects reported that wound healing was significantly accelerated in the first month of applying the ointment, and 34 subjects reported that their skin scars were significantly softer and smaller, and the color of the scars gradually faded after two months of using the ointment. The condition of the scars was significantly improved.

[0086] Example 7: Skin Care Repair Lotion The formula for the Skin Care Repair Lotion is shown in the table below:

[0087] [Table 6]

[0088] The preparation method for the above skin care repair lotion was as follows: (1) Add aloe extract, honey and cetyl glucoside to a reactor, add half of the 2x concentrated birch sap, heat to 75°C, and dissolve with stirring for 30 minutes to obtain solution A; (2) Add pearl powder, milk powder and lemon extract to a container, add the remaining half of the double-concentrated birch sap, stir evenly, heat to 75°C, and stir for 10 minutes to dissolve, obtaining solution B; (3) Solution B was slowly added to solution A and homogenized at 5000 rpm for 3 minutes. When the temperature dropped to 70°C, bis(hydroxymethyl)imidazolidinyl urea, carbomer, antioxidant and emulsifier were added, and the mixture was stirred at 200 rpm for 1 hour, cooled to room temperature and packaged to obtain a skin care repair lotion.

[0089] Twenty subjects were treated with the above skin care repair lotion for two months and then subjectively evaluated. The results showed that 18 subjects reported that after applying the lotion, the elasticity of the skin on the scar surface increased, the scar pigment weakened, and the scar area decreased, demonstrating the scar repair effect.

[0090] The technical solutions in the above examples were preferred embodiments of the present invention. Some improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered to fall within the protection scope of the present invention.

Claims

1. 1. A topical skin composition comprising: (A) the composition for topical use on skin contains 30 to 98% by weight of concentrated birch sap based on the total weight of the composition, and the concentrated birch sap has a concentration of 1.5 to 5 times; (B) further comprising 0.001 to 10 wt. % of one or more active substances selected from Coptis japonica root extract, Lithospermum officinalis extract, allantoin, Salvia miltiorrhiza extract, Astragalus root extract, and asiaticoside, based on the total weight of the topical skin composition; The topical skin composition does not contain any additional water, but does not exclude the water inherently contained in each of all components contained in the composition.

2. 10. The topical skin composition of claim 1, wherein the concentrated birch sap has a concentration of 1.5 to 3 times.

3. 3. The topical skin composition of claim 1, wherein the total content of component (B) in the topical skin composition is 0.1 to 5%.

4. 4. The topical skin composition of claim 3, wherein the total content of component (B) in the topical skin composition is 0.5 to 3%.

5. 3. The topical skin composition of claim 1 or 2, comprising a pharmaceutical composition and a cosmetic skin care composition.

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