Composition for wound healing comprising sesamol and use thereof

Abstract

The present invention relates to a composition for wound healing comprising sesamol, and a use thereof, and more particularly, to a pharmaceutical composition or cosmetic composition comprising a low concentration of sesamol for skin regeneration, wound healing, or an antibacterial purpose. In the present invention, by using a low concentration of sesamol without an additional synthesis process, a cream-based composition for wound healing, which compensates for existing shortcomings and maximizes inherent healing effects, has been developed. In addition, it was confirmed that a high concentration of sesamol rather decreases wound healing effects, whereas a composition including sesamol in an amount less than 1% (w / w) has the effect of maximizing the wound healing effects and minimizing scars by acting in a complex manner throughout the entire wound healing process. Thus, the sesamol in an amount less than 1% (w / w) can be effectively applied to the composition for wound healing or the composition for skin regeneration.

Description

Wound healing composition containing sesamol and uses thereof

[0001] The present invention relates to a wound healing composition containing sesamol and its uses, and more specifically, to a pharmaceutical composition or cosmetic composition for skin regeneration, wound healing, or antibacterial purposes containing a low concentration of sesamol.

[0002]

[0003] The skin performs various functions in our body, the most important of which is its role as a barrier protecting us from the outside. A wound is defined as a condition where the normal structure of the skin is compromised. When the skin is injured in this way, the wound site becomes exposed, making it susceptible to the invasion of external bacteria. Therefore, prompt wound treatment is essential to prevent further infection and to stop excessive inflammatory responses. Excessive inflammation not only prolongs the wound healing period but can also lead to scarring.

[0004] Wound healing clinically refers to the complete closure of the skin, and general wounds, excluding chronic wounds, heal completely within approximately 3 to 14 days. Wound healing is a complex process regulated by the interaction of various types of cells, including keratinocytes, fibroblasts, endothelial cells, macrophages, and platelets.

[0005] Steroids control skin inflammation through immunosuppression, but they delay wound healing by inhibiting the proliferation of keratinocytes or inhibiting cell proliferation. As such, if wound healing is delayed due to steroids, the risk of secondary infection and other complications increases, so the development of wound healing agents remains necessary.

[0006]

[0007] Meanwhile, sesamol is a natural phenolic compound extracted from sesame seeds that has powerful antioxidant, anti-inflammatory, and antibacterial effects, making it useful for promoting wound healing. However, although sesamol possesses excellent antioxidant, anti-inflammatory, and antibiotic effects, due to the characteristic foul odor of phenols, it is essential to include development stages primarily for derivatives (Korean Patent Publication No. 10-2006-0130835).

[0008]

[0009] In this invention, as a result of diligent efforts to apply sesamol for wound treatment without an additional synthesis process, a cream formulation composition containing a low concentration of sesamol was prepared. It was confirmed that the low-concentration sesamol cream of this invention fully preserves the inherent properties of sesamol, thereby maximizing wound healing and skin protection effects and minimizing the possibility of scar formation based on high-efficiency control of reactive oxygen species levels and multifunctionality. Furthermore, the sesamol wound healing cream of this invention is characterized by having a high moisture content through an emulsification process at an intermediate temperature, which minimizes scarring that occurs in dry environments and maximizes the wound healing effect.

[0010]

[0011] Accordingly, the objective of the present invention is to provide a pharmaceutical composition or cosmetic composition for wound treatment comprising a low concentration of sesamol.

[0012] Another objective of the present invention is to provide an antibacterial pharmaceutical composition or cosmetic composition comprising a low concentration of sesamol.

[0013]

[0014] In order to achieve the aforementioned purpose,

[0015] The present invention provides a pharmaceutical composition for regeneration after skin damage, wound treatment, or prevention comprising less than 1% (w / w) of sesamol as an active ingredient.

[0016] The present invention provides a cosmetic composition for skin regeneration, skin protection, or wound improvement comprising less than 1% (w / w) of sesamol as an active ingredient.

[0017] The present invention provides an antimicrobial pharmaceutical composition comprising less than 1% (w / w) of sesamol as an active ingredient.

[0018] The present invention provides an antibacterial cosmetic composition comprising less than 1% (w / w) of sesamol as an active ingredient.

[0019] The present invention provides a wound treatment method using less than 1% (w / w) of sesamol.

[0020]

[0021] In a preferred embodiment of the present invention, the sesamol may be included in the composition at 0.1% (w / w) or less, preferably at 0.03% (w / w) to 0.08% (w / w), and more preferably at 0.04% (w / w) to 0.06% (w / w).

[0022] In another preferred embodiment of the present invention, the composition may further comprise stearic acid, cetostearyl alcohol, mineral oil, glycerin and / or triethanolamine, and preferably may further comprise 0.04% (w / w) to 0.06% (w / w) stearic acid, 0.02% (w / w) to 0.05% (w / w) cetostearyl alcohol, 0.04% (w / w) to 0.06% (w / w) mineral oil, 0.04% (w / w) to 0.06% (w / w) glycerin and / or 0.005% (w / w) to 0.006% (w / w) triethanolamine based on the total composition.

[0023] In another preferred embodiment of the present invention, the composition may additionally include one or more components selected from the group consisting of propyl paraben, methyl paraben, and tocopherol as preservatives.

[0024] In another preferred embodiment of the present invention, the composition may be in the form of a cream, emulsion, ointment, or lotion.

[0025]

[0026] In the present invention, a wound healing cream composition was developed that overcomes existing disadvantages and maximizes unique therapeutic effects by utilizing a low concentration of sesamol without an additional synthesis process. Furthermore, while high concentrations of sesamol actually reduced the wound healing effect, it was confirmed that a composition containing less than 1% (w / w) of sesamol acts in combination during the pre-wound healing stage to maximize wound healing effects and minimize scarring effects, so it can be effectively applied to wound healing compositions or skin regeneration compositions.

[0027]

[0028] Figure 1 shows the cytotoxicity of sesamol in HaCaT (immortalized human keratinocyte cell line) cells and NIH-3T3 (a fibroblast cell line that was isolated from a mouse NIH / Swiss embryo).

[0029] Figure 2 is data confirming the ability of sesamol to remove active oxygen.

[0030] Figure 3 is data confirming the cell proliferation ability by high concentration sesamol.

[0031] Figure 4 is data confirming the cell proliferation ability by low concentration sesamol.

[0032] Figure 5 is data confirming the antibacterial ability of Sesamol.

[0033] Figure 6 shows data confirming the wound healing ability of sesamol, Figure 6a shows data confirming the wound healing ability using high concentration of sesamol, and Figure 6b shows data confirming the wound healing ability using low concentration of sesamol.

[0034] Figure 7 is an image confirming the formulation of the manufactured Sesamol wound healing cream.

[0035] Figure 8 shows the acidity of Sesamol wound healing cream.

[0036] Figure 9 is an image confirming the skin protective effect of Sesamol wound healing cream.

[0037] Figure 10 shows data confirming the wound healing effect of Sesamol wound healing cream in an animal model, Figure 10a is a photograph observing the wound size after applying Sesamol wound healing cream, and Figure 10b is data quantifying the wound size.

[0038] Figure 11 is data obtained by analyzing tissue to confirm the wound healing effect of Sesamol wound healing cream in an animal model.

[0039]

[0040] The present invention will be described in detail below.

[0041]

[0042] In one aspect, the present invention relates to a pharmaceutical composition for the regeneration of skin after damage, wound treatment, or prevention comprising less than 1% (w / w) of sesamol as an active ingredient.

[0043] In another aspect, the present invention relates to a cosmetic composition for skin regeneration, skin protection, or wound improvement comprising less than 1% (w / w) of sesamol as an active ingredient.

[0044]

[0045] In the present invention, the term "active ingredient" refers to a component that exhibits the intended activity alone or can exhibit the intended activity in combination with a carrier, etc., which is inactive itself.

[0046] In the present invention, the term "treatment" refers to any act in which symptoms such as infection by bacteria, skin damage, or wounds are improved, eliminated, or benefited by the administration (including application) of the sesamol according to the present invention.

[0047] The term "prevention" in the present invention refers to any act that can inhibit or delay the progression of infection by bacteria, skin damage, or wounds by administering (including application) sesamol according to the present invention.

[0048] The term "improvement" in the present invention refers to any act in which symptoms such as infection by bacteria, skin damage, or wounds are improved or beneficially altered by administering sesamol according to the present invention.

[0049]

[0050] In the present invention, sesamol may be included in the composition at 0.1% (w / w) or less, preferably at 0.03% (w / w) to 0.08% (w / w), and more preferably at 0.04% (w / w) to 0.06% (w / w).

[0051] In the present invention, the composition may further comprise stearic acid, cetostearyl alcohol, mineral oil, glycerin and / or triethanolamine, and preferably, with respect to the total composition, may further comprise 0.04% (w / w) to 0.06% (w / w) stearic acid, 0.02% (w / w) to 0.05% (w / w) cetostearyl alcohol, 0.04% (w / w) to 0.06% (w / w) mineral oil, 0.04% (w / w) to 0.06% (w / w) glycerin and / or 0.005% (w / w) to 0.006% (w / w) triethanolamine.

[0052] In the present invention, the composition may additionally include one or more components selected from the group consisting of propyl paraben, methyl paraben, and tocopherol as preservatives.

[0053]

[0054] Sesamol has excellent antioxidant, anti-inflammatory, and antibiotic effects, but due to the characteristic odor of phenols, it is essential to include development stages primarily for derivatives. However, in the present invention, a cream formulation was created using low concentrations of sesamol without additional synthesis processes, thereby compensating for existing disadvantages and maximizing the unique therapeutic effect.

[0055] In a specific embodiment of the present invention, the cytotoxicity and proliferative capacity of sesamol were evaluated on fibroblasts and keratinocytes, and it was confirmed that sesamol possesses high biocompatibility and the ability to promote cell proliferation. This implies that sesamol can shorten the wound healing period and minimize side effects such as infection.

[0056] In addition, in an evaluation of the efficiency of removing reactive oxygen species using hydrogen peroxide, a major reactive oxygen species generated in wound sites, sesamol demonstrated excellent antioxidant ability by inhibiting the level of reactive oxygen with high efficiency ranging from about 60% to 98% at low to high concentrations, and an average of over 80% based on the lowest and highest concentrations.

[0057]

[0058] Regarding sesamol concentration, it has been reported that high concentrations of sesamol of 500 μM (approx. 70 μg / ml) or higher inhibit autophagosome formation and induce apoptosis (Zhigang Liu, et. al., Scientific Reports, 7:45728, 2017); therefore, high concentrations of sesamol refer to concentrations of 500 μM or higher. Since the low concentration of sesamol defined in this invention is an effective concentration that has been confirmed to exhibit effective wound healing efficacy without cytotoxicity, the low concentration of sesamol in this invention can be viewed as a differentiated concentration setting proven by existing research results.

[0059]

[0060] Based on the above, in another specific embodiment of the present invention, a wound healing cream containing low concentration sesamol was prepared, and in a wound healing model established using keratinocytes, low concentration sesamol showed a wound healing effect of about 20% or more compared to the control group, and from this, it was confirmed that sesamol provides complex functionality in wound healing.

[0061] Furthermore, the wound healing cream containing low concentrations of sesamol showed excellent skin protective effects within the pH range that maintains biocompatibility, and demonstrated significant wound healing effects (P≤ 0.05 and P≤ 0.01) on days 7 and 9, respectively, in an animal model of wound healing.

[0062]

[0063] It has been confirmed that the sesamol wound healing cream of the present invention possesses a high moisture content through an emulsification process at an intermediate temperature, which can maximize the wound healing effect by minimizing scarring that occurs in dry environments, and that by fully preserving the inherent properties of sesamol, it maximizes wound healing and skin protection effects and minimizes the possibility of scar formation based on high-efficiency control of active oxygen levels and multifunctionality.

[0064] In other words, the present invention compensates for the disadvantages of the raw material by using a wound healing cream containing a low concentration of sesamol without an additional synthesis process, and develops a functional wound healing cream that simultaneously possesses high-efficiency active oxygen scavenging function and skin protective effect.

[0065]

[0066] The pharmaceutical composition of the present invention may be a pharmaceutical composition for external use on the skin and may be formulated into various forms according to conventional methods. Preferably, the composition may be in the form of a cream, emulsion, ointment, or lotion. Furthermore, in each formulation, other components of the composition of the present invention may be arbitrarily selected and combined according to the formulation or purpose of use. The amount of the active ingredient may be appropriately determined according to the purpose of use (inhibition or alleviation). For example, it may include conventional adjuvants such as thickeners, stabilizers, solubilizers, vitamins, pigments, and fragrances, and carriers.

[0067]

[0068] In another aspect, the present invention relates to a quasi-drug composition for the regeneration of skin after damage, wound treatment, or prevention comprising less than 1% (w / w) of sesamol as an active ingredient.

[0069] In the present invention, "quasi-drug" refers to articles used for the purpose of diagnosing, treating, improving, alleviating, managing, or preventing diseases of humans or animals, among which the effect is milder than that of pharmaceuticals. For example, quasi-drugs under the Pharmaceutical Affairs Act are those excluding articles used for pharmaceutical purposes, and include products used for the treatment or prevention of diseases in humans / animals, products that have a mild effect on the human body or do not act directly on it, etc.

[0070]

[0071] In the cosmetic composition of the present invention, the cosmetic may be an ampoule, cream, lotion, toner, essence, or pack, but is not necessarily limited thereto.

[0072] The cosmetic composition according to the present invention may be composed of, in addition to the above-mentioned sesamol, various ingredients generally used in cosmetic compositions, such as water-soluble ingredients, powder ingredients, oils, surfactants, moisturizers, viscosity modifiers, preservatives, antioxidants, fragrances, colorants, etc., as needed and within a range that does not reduce the effect of the present invention.

[0073] In another aspect, the present invention relates to an antimicrobial pharmaceutical composition comprising less than 1% (w / w) of sesamol as an active ingredient.

[0074] In another aspect, the present invention relates to an antibacterial cosmetic composition comprising less than 1% (w / w) of sesamol as an active ingredient.

[0075] In another aspect, the present invention relates to a method for treating skin damage and / or treating wounds, comprising the step of administering or applying less than 1% (w / w) of sesamol to an individual.

[0076]

[0077] The present invention will be explained in more detail below through examples.

[0078] These examples are solely for illustrating the invention, and it will be obvious to those skilled in the art that the scope of the invention is not to be interpreted as being limited by these examples.

[0079]

[0080] Example 1: Confirmation of skin cytotoxicity of sesamol

[0081] In the present invention, the cytotoxicity of sesamol was evaluated by selecting HaCaT (immortalized human keratinocyte cell line) and NIH-3T3 (a fibroblast cell line that was isolated from a mouse NIH / Swiss embryo) as two representative cell lines constituting the skin for wound healing. Keratinocytes and fibroblasts are representative cells constituting the epidermis and dermis of the skin, and since they are the first to suffer severe damage when a wound occurs, they are cell lines generally used when evaluating developed wound healing agents in vitro.

[0082] Specifically, 1 x 10 4 Cells were seeded into 96-well plates and cultured for 24 hours at 37°C under 5% CO2 conditions. Sesamol (molecular weight = 138 g / mol) was diluted by serial dilution to a maximum concentration of 1 mg / ml, and after treatment with each cell, the cells were cultured for 24 hours, after which toxicity was evaluated using the CCK-8 cytotoxicity kit (Cell Counting Kit-8, Dojindo).

[0083]

[0084] As a result, as shown in Figure 1, it was confirmed that cell viability of over 80% was observed even at a concentration of 250 μg / ml of sesamol. In addition, it was confirmed that low concentrations of sesamol, ranging from 7.8 to 31.25 μg / ml, showed cell proliferation rates similar to or higher than those of the control group (0 μg / ml) that was not treated with sesamol.

[0085]

[0086] Example 2: Confirmation of Sesamol's Reactive Oxygen Scavenging Ability

[0087] In this invention, the reactive oxygen species scavenging ability of sesamol was evaluated using hydrogen peroxide (H2O2), a major reactive oxygen species (ROS). Specifically, sesamol solutions at concentrations ranging from 0 to 500 μg / ml were treated with a constant concentration of 50 μM hydrogen peroxide and incubated at room temperature for 30 minutes, after which the fluorescence signal was quantified using an H2O2 assay kit (λ exc 520 nm, λ emi 580 ~ 610 nm).

[0088]

[0089] As a result, as shown in Figure 2, excellent reactive oxygen species scavenging ability was confirmed starting from the lowest concentration of sesamol. The hydroxyl group (-OH) of sesamol plays a role in accepting and stabilizing free radicals of reactive oxygen species. Consequently, the hydrogen atom of the hydroxyl group of sesamol is removed, and the formed sesamol radical forms a sesamol dimer for stabilization. Reactive oxygen species are removed in the order of radical neutralization, stabilization, and dimer formation. Sesamol's low molecular weight allows it to penetrate the cell membrane with high solubility, making it effective for removing reactive oxygen species within the cell.

[0090]

[0091] In addition, the scavenging ability of sesamol against DPPH, a reactive oxygen species and a type of free radical, was evaluated. Specifically, sesamol solutions at concentrations ranging from 0 to 500 μg / ml were treated with a constant concentration of DPPH (50 μM) and incubated at room temperature for 10 minutes, and the absorbance based on color change was quantified using a DPPH assay kit (λ abs 517 nm).

[0092] As a result, as shown in Figure 2, it was confirmed that a significant decrease was observed starting from the lowest concentration of 7.81 μg / ml (EC 50 = 250 µg / ml).

[0093]

[0094] Example 3: Confirmation of Sesamol's Cell Proliferation Ability

[0095] One of the essential elements required for wound healing is the proliferative capacity of cells that make up the skin. Based on the results of the cytotoxicity evaluation in <Example 1> above, the cell proliferation capacity of two types of cell lines was evaluated using a method that evaluates cell proliferation capacity using a CCK-8 assay kit and Alamar blue with sesamol at a concentration of 0 to 500 μg / ml.

[0096] As a result, as shown in Figure 3, on the first day of culture in keratinocytes (HaCaT), a proliferation ability approximately 1.17 to 1.12 times better than the control group was observed at a sesamol concentration of 50 to 100 μg / ml, but it was confirmed that it did not have a significant effect on cell proliferation on the second and third days.

[0097] In the case of high concentration sesamol, it was observed that it did not significantly affect cell proliferation or even decreased from the second day, so the cell proliferation ability in the low concentration group of 0 to 30 μg / ml was additionally evaluated.

[0098] As a result, as shown in Figure 4, on the first day of culture in keratinocytes (HaCaT), a proliferation ability improved by about 1.02 to 1.1 times compared to the control group was observed at a sesamol concentration of 5 to 20 μg / ml, on the second day, a cell proliferation rate of more than 1.9 times was observed at 5 μg / ml sesamol, and on the third day, it was confirmed that it did not have a significant effect on cell proliferation.

[0099]

[0100] Example 4: Evaluation of the antimicrobial activity of sesamol

[0101] In this invention, a minimum inhibitory concentration (MIC) test was performed to evaluate the inherent antibacterial ability of sesamol.

[0102] It has already been reported that sesamol has an antibacterial effect, but only against some 2 to 3 types of bacteria including E. coli. We wanted to perform additional verification to see if sesamol could serve as a tool to (1) prevent product spoilage while using the product and (2) prevent secondary infection in wounds when a cream containing sesamol is manufactured.

[0103] For the MIC test, eight types of bacteria were selected, including those known as representative hospital-acquired pathogens and those that cause secondary infections in wounds. Staphylococcus aureus (SA) and Methicillin-resistant Staphylococcus aureus (MRSA) are Gram-positive bacteria, while the remaining bacteria are Gram-negative. The bacteria used for evaluation were selected based on the representative hospital-acquired pathogens announced by the National Institutes of Health and the strains most statistically isolated from wounds.

[0104]

[0105] MIC applied strainBacteriaMICE. coli(EC)2.5 mg / mLA. baumannii(AB)1.25 mg / mLP. Aeruginosa(PA)5 mg / mLP. Mirabilis(PM)1.25 mg / mLK. Pneumoniae(KP)2.5 mg / mLE. Faceium(EF)5 mg / mLS. aureus(SA)1.25 ㎎ / ㎖Methicillin-resistanceS. aureus(MRSA)5 mg / ml

[0106] The concentrations of sesamol used for the MIC test ranged from 0.625 to 10 mg / mL. Based on the maximum concentration, the sesamol was serially diluted in CAMHB (Cation-adjusted Mueller Hinton Broth) medium and dispensed into 96-well plates. Specifically, bacteria were streaked onto LB Agar and serum media, respectively, and cultured for 18 hours at 37°C. A single colony was then collected and diluted in sterile distilled water to prepare a McFarland 0.5 standard turbidity solution. This solution was then inoculated into 96-well plates at a 1 / 10 dilution ratio and cultured for approximately 18 hours at 37°C. The negative control (NC) consisted only of the medium, while the positive control (PC) consisted only of the medium and bacteria.

[0107] As a result, as shown in Figure 5, the same experimental group was divided into Round-bottom (left photo) and Flat-bottom well plates (right graph) for analysis. In each bacterial group, at a concentration of 1.25 to 2.5 mg / ml, the antibacterial effect was observed against most skin infection bacteria, and at a higher concentration of 5 mg / ml, an antibacterial effect was observed even against methicillin-resistant Staphylococcus aureus and bacteria mainly found in chronic wound infections.

[0108] Verification at the cellular level was based on the therapeutic effect when sesamol actually enters the cell, so the concentration was measured in μg / ml units; however, in this case, considering the concentration of sesamol included in the cream during manufacturing, verification was performed in mg / ml units. In conclusion, it was confirmed that sesamol is a functional ingredient for wound healing and can also help maintain the state of the cream through its antibacterial ability.

[0109]

[0110] Example 5: Evaluation of Sesamol's Wound Healing Ability

[0111] In this invention, the wound healing ability of sesamol was evaluated using HaCaT cells, which are keratinocytes located on the outermost layer of the skin.

[0112]

[0113] Specifically, attach the Wound Healing Assessment Kit (ibidi Wound Healing and Migration Assay Kit, Culture-Insert 2 Well) to a 24-well plate and 5 x 10 5 Approximately 70 µl of DMEM (Dulbecco's Modified Eagle Medium) containing HaCaT cells was dispensed into both wells of the insert and cultured for 24 hours at 37°C under 5% CO2 conditions. Then, after removing the insert, fresh 1 ml of DMEM (containing 10% FBS and 1% penicillin-streptomycin) medium was added to each well, followed by treatment with sesamol solution at concentrations ranging from 0 to 2000 µg / ml. The concentration of the treated sesamol solution refers to the final treatment concentration considering the volume of the medium.

[0114]

[0115] As a result of comparing the wound healing effects between the group not treated with sesamol and the group treated with sesamol for 0 to 24 hours, as shown in Figure 6a, it was confirmed that there was a statistically significant wound healing effect between 0 μg / ml sesamol (control group) and 20 to 40 μg / ml sesamol at 24 hours. In addition, a Live & Dead assay was performed to clearly observe the wound healing effect visually, and the same pattern was confirmed.

[0116] It was confirmed that the wound healing effect was superior in low concentrations of sesamol compared to high concentrations of sesamol, and a low concentration of sesamol solution with a concentration of 0 to 30 μg / ml was applied.

[0117] As a result of comparing the wound healing effect between a group not treated with sesamol and a group treated with sesamol for 0 to 24 hours, as shown in Figure 6b, it was confirmed that there was a statistically significant wound healing effect between 0 μg / ml sesamol (control group) and 5 to 10 μg / ml sesamol at 24 hours. In addition, a Live & Dead assay was performed to clearly observe the wound healing effect visually, and the same pattern was confirmed. In other words, the present invention confirmed that the wound healing effect of low concentrations of sesamol is excellent.

[0118]

[0119] Example 6: Preparation of Sesamol wound healing cream

[0120] Generally, considering that wound healing agents are applied topically multiple times over a short period, a wound healing cream was prepared within the low concentration range. Based on the results of cytotoxicity, cell proliferation, and reactive oxygen species scavenging ability, the factors considered for setting the concentration were to prepare a wound healing cream with a sesamol concentration of 0.05% (w / w), which is within the range of 20 μg / ml per single application, and as comparative examples, creams were prepared by including groups with a concentration 20 times higher (1%) and a concentration 100 times higher (5%).

[0121]

[0122] Sesamol Wound Healing Cream Ingredients and Content Ingredient Content (Weight%) Stearic Acid 5 g (0.05%) Cetostearyl Alcohol 3.6 g (0.036%) Mineral Oil 5 g (0.05%) Glycerin 5 g (0.05%) Triethanolamine 0.56 g (0.0056%) Sesamol 0.05-0.4 g (0.05-5%) Purified Water ~100 g (~100%)

[0123] A wound healing cream composition was prepared as shown in the composition of Table 2 above, and was prepared to include the lipid-soluble components stearic acid, mineral oil, cetostearyl alcohol (cetearyl alcohol), which is an emulsifying stabilizer and surfactant, and triethanolamine. Since sesamol has an antibacterial effect in the wound healing cream composition, there is no need to add synthetic compounds known as preservatives, but depending on the purpose, 0.0001 to 0.0003% (w / w) propyl paraben, 0.0005 to 0.0025% (w / w) methyl paraben, and 0.003 to 0.005% (w / w) tocopherol may be additionally included.

[0124]

[0125] Sesamol wound healing cream was formulated by continuously using a homogenizer while heating the water-soluble and oil-soluble components to conditions of 70 to 80°C, respectively. Specifically, 50 ml of purified water was added to glycerin, the water-soluble component (A), and homogenized under heated conditions. Then, the oil-soluble component (B), excluding the active ingredient sesamol, was homogenized at the same temperature until it was completely dissolved, at which point the two components were mixed to form an emulsion. The cream was produced by increasing the moisture content at the same temperature until it stabilized. Then, the mixture was cooled to room temperature (25°C), the active ingredient sesamol was added, and the mixture was further homogenized for about 30 minutes to 1 hour to produce the Sesamol wound healing cream. The formulation and color of the manufactured Sesamol wound healing cream are shown in Fig. 7. The cream base refers to the basic formulation of the Sesamol cream in which all components, excluding the active ingredient sesamol, are included in equal amounts.

[0126]

[0127] In addition, the acidity (pH) of the cream base and Sesamol cream was measured using biocompatible PBS with a pH of 7.4 as a control, and as shown in Figure 8, both showed a pH within the biocompatible range. Specifically, 0.5 g of the cream was dissolved in 10 ml of PBS, and the pH was evaluated repeatedly at least three times using an electronic pH meter.

[0128]

[0129] Example 7: Confirmation of the skin protective effect of Sesamol wound healing cream

[0130] To verify the skin protective effect of a wound healing cream containing sesamol, the cream was evaluated using filter paper coated with the cream. Phenolphthalein is a representative pH indicator characterized by exhibiting a color change in a strongly alkaline environment. Potassium hydroxide (KOH) is a strongly alkaline solution, and the protective ability is evaluated based on the purple color development of the phenolphthalein-coated paper when the cream has no protective ability.

[0131] Specifically, a 1% phenolphthalein ethanol solution was prepared and coated once onto a 7 x 7 cm filter paper, which was then dried in a 37°C oven for 1 hour. 0.5 g of the prepared 0.05% (w / w) sesamol wound healing cream was applied to a 2 x 2 cm filter paper and placed on top of the phenolphthalein-coated paper. After applying 20 µl of 0.1 N KOH once, the immediate skin protective effect was observed for 0 to 30 minutes. The evaluation groups included an untreated control group, a cream base, and a sesamol cream; skin protective effects were observed in all experimental groups except the control group (Fig. 9).

[0132]

[0133] Example 8: Confirmation of wound healing effect by Sesamol wound healing cream in an animal model

[0134] In this invention, the wound healing effect of Sesamol wound healing cream was to be confirmed in an animal model.

[0135] First, wounds were created in a mouse animal model using a 6 mm biopsy punch, and the size of the wounds was measured at 3, 5, 7, 9, 11, and 13 days in a control group that received only daily alcohol disinfection, and groups that received alcohol disinfection followed by cream, or 0.05% (w / w), 1% (w / w), or 5% (w / w) Sesamol wound healing cream.

[0136] As a result, as shown in Figures 10a and 10b, it was confirmed that the group treated with 0.05% (w / w) Sesamol wound healing cream recovered faster than the control group on days 7 and 9. In other words, it can be seen that the reactive oxygen species and immune response generated upon injury are improved by the antioxidant and anti-inflammatory effects of Sesamol wound healing cream, thereby confirming that low concentrations of Sesamol wound healing cream are effective for wound treatment.

[0137]

[0138] On the other hand, it was observed that the wound healing effect decreased as the concentration of sesamol increased. In fact, in Example 1 above, it was confirmed that cell proliferation was inhibited or toxicity was exhibited when the concentration of sesamol was above a certain level. While the 1% (w / w) sesamol wound healing cream showed superior recovery ability compared to the cream base, the 5% (w / w) sesamol wound healing cream was judged to have a reduced healing effect compared to the base because it was a high concentration that inhibited the proliferation rate.

[0139]

[0140] Since various factors interact in the wound healing process, the degree of recovery at the tissue level is important in addition to the size of the wound visible to the naked eye; therefore, mouse tissues from the group treated with the most effective 0.05% (w / w) Sesamol wound healing cream were obtained to evaluate the extent of wound healing at the tissue level. Histological evaluation was performed using Masson trichrome (MT) staining, in which muscle tissue is stained red, collagen fibers blue, and cell nuclei black. Additionally, collagen was quantified across the groups using MT staining.

[0141] As a result, as shown in Figure 11, a statistically significant reduction in collagen brightness and % area was observed in the 0.05% (w / w) sesamol wound healing cream group compared to the control group. On the other hand, no decreasing trend was observed in the creams containing 1% and 5% sesamol compared to the control group.

[0142] Collagen increases due to the initial immune response after a wound occurs, but it should decrease as the wound heals. If collagen is not reduced and remains constant, the wound can develop into a scar. The decrease in collagen in the group treated with 0.05% (w / w) Sesamol wound healing cream means that complete skin regeneration can occur without the wound developing into a scar.

[0143]

[0144] In the present invention, a wound healing cream composition was developed that overcomes existing disadvantages and maximizes unique therapeutic effects by utilizing a low concentration of sesamol without an additional synthesis process. Furthermore, while high concentrations of sesamol actually reduced the wound healing effect, it was confirmed that a composition containing less than 1% (w / w) of sesamol acts in combination during the pre-wound healing stage to maximize wound healing effects and minimize scarring effects, so it can be effectively applied to wound healing compositions or skin regeneration compositions.

Claims

A pharmaceutical composition for regeneration after skin damage, wound treatment, or prevention, comprising less than 1.1% (w / w) of sesamol as an active ingredient.

2. In Paragraph 1, A pharmaceutical composition for regeneration after skin damage, wound treatment, or prevention, characterized in that the above composition contains 0.1% (w / w) or less of sesamol.

3. In Paragraph 1, A pharmaceutical composition for regeneration after skin damage, wound treatment, or prevention, characterized in that the above composition further comprises one or more components selected from the group consisting of stearic acid, cetostearyl alcohol, mineral oil, glycerin, and triethanolamine.

4. In Paragraph 1, A pharmaceutical composition for regeneration after skin damage, wound treatment, or prevention, characterized in that the above composition further comprises, based on the total composition, 0.04% (w / w) to 0.06% (w / w) stearic acid, 0.02% (w / w) to 0.05% (w / w) cetostearyl alcohol, 0.04% (w / w) to 0.06% (w / w) mineral oil, 0.04% (w / w) to 0.06% (w / w) glycerin, and 0.005% (w / w) to 0.006% (w / w) triethanolamine.

5. In Paragraph 1, A pharmaceutical composition for regeneration after skin damage, wound treatment, or prevention, characterized in that the above composition is in the form of a cream, emulsion, ointment, or lotion. A cosmetic composition for skin regeneration, skin protection, or wound improvement, comprising less than 6.1% (w / w) of sesamol as an active ingredient.

7. In Paragraph 6, A cosmetic composition for skin regeneration, skin protection, or wound improvement, characterized in that the above composition contains 0.1% (w / w) or less of sesamol.

8. In Paragraph 6, A cosmetic composition for skin regeneration, skin protection, or wound improvement, characterized in that the above composition further comprises one or more components selected from the group consisting of stearic acid, cetostearyl alcohol, mineral oil, glycerin, and triethanolamine.

9. In Paragraph 6, A cosmetic composition for skin regeneration, skin protection, or wound improvement, characterized in that the above composition further comprises, based on the total composition, 0.04% (w / w) to 0.06% (w / w) stearic acid, 0.02% (w / w) to 0.05% (w / w) cetostearyl alcohol, 0.04% (w / w) to 0.06% (w / w) mineral oil, 0.04% (w / w) to 0.06% (w / w) glycerin, and 0.005% (w / w) to 0.006% (w / w) triethanolamine.

10. In Paragraph 6, A cosmetic composition for skin regeneration, skin protection, or wound improvement, characterized in that the above composition is in the form of a cream, emulsion, ointment, or lotion. A quasi-drug composition for regeneration after skin damage, wound treatment, or prevention, comprising less than 11.1% (w / w) of sesamol as an active ingredient. An antimicrobial pharmaceutical composition comprising less than 12.1% (w / w) of sesamol as an active ingredient. Antibacterial cosmetic composition comprising less than 13.1% (w / w) of sesamol as an active ingredient.

14. A wound treatment method comprising the step of administering or applying less than 1% (w / w) of sesamol to an individual.

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