Application of alpha-ketobutyrate in preparing skin care products and medicines for preventing and or treating skin photoaging

Abstract

The present invention discloses the use of α-ketobutyrate in the preparation of skin care products and drugs for preventing and or treating skin photoaging. Balb / c mice are used to construct a skin photoaging mouse model using a sunlight simulator. α-ketobutyrate is administered for intervention. The results show that α-ketobutyrate intervention can significantly improve the pathological phenotypes such as a large number of erythema, wrinkles, desquamation, inflammation and thickening of the stratum corneum presented by the skin of the photoaging model mice. α-ketobutyrate intervention can significantly increase the expression of E-Cadherin, ZO-1 and Collagen I in the skin tissue of the photoaging model mice. α-ketobutyrate intervention can significantly reduce the expression of DNA damage and p53 and p21 aging markers in the skin tissue of the photoaging model mice. At the same time, α-ketobutyrate is an endogenous metabolite with strong pharmacological effects and small side effects, thus providing a basis for the clinical development of skin care products and drugs for the treatment of skin photoaging.

Description

Technical Field

[0001] The present invention belongs to the technical field of skin disease biomedicine, and particularly relates to the use of α-ketobutyrate in the preparation of skin care products and medicines for preventing and or treating skin photoaging. Background Art

[0002] Skin, the largest organ in the human body, is inevitably exposed to the external environment. Over time, and due to the accumulation of various stimuli, it undergoes natural aging and photoaging. Skin photoaging is damage to the skin caused by long-term exposure to sunlight. It manifests as roughness, thickening, and dryness in exposed areas of the skin, sagging skin, deeper and coarser wrinkles, localized excessive pigmentation or capillary dilation, and even the development of various benign or malignant tumors (such as solar keratosis, squamous cell carcinoma, and malignant melanoma). Skin photoaging and light damage are currently the most common skin diseases in the world and have become a global health issue.

[0003] Despite steady progress in recent years in elucidating the pathogenesis of photoaging skin diseases, identifying therapeutic targets, and advancing drug development, significant unresolved challenges remain. Over the past few decades, basic and translational research has advanced our understanding of the pathogenesis of photoaging skin diseases and identified several potential therapeutic targets. Currently, the main treatments for photoaging skin diseases include sunscreen, oral or topical tretinoin preparations, moisturizing, sunscreen, and antioxidants, and phototherapy. However, these treatments, while primarily preventative or physical, often face the body's own immune responses and significant side effects, potentially hindering their clinical application. Summary of the Invention

[0004] The purpose of the present invention is to overcome the shortcomings of the prior art and provide the use of α-ketobutyrate in the preparation of skin care products and medicines for preventing and or treating skin photoaging. α-ketobutyrate is an endogenous metabolite in the human body that can eliminate autoimmune reactions and has relatively few side effects. The targets are mainly DNA damage and aging markers. α-ketobutyrate has great prospects in the clinical treatment of photoaging skin diseases.

[0005] The technical solution to achieve the above purpose is: use of α-ketobutyrate in the preparation of skin care products and medicines for preventing and or treating skin photoaging, wherein the structural formula of the α-ketobutyrate is:

[0006]

[0007] The α-ketobutyrate is used in the preparation of skin care products and medicines for preventing and or treating skin photoaging, wherein the α-ketobutyrate improves erythema, wrinkles, desquamation, inflammation and stratum corneum thickening on the skin of mice.

[0008] The α-ketobutyrate is used in the preparation of skin care products and medicines for preventing and or treating skin photoaging, wherein the α-ketobutyrate increases the expression of E-Cadherin protein in mouse skin tissue.

[0009] The α-ketobutyrate is used in the preparation of skin care products and medicines for preventing and or treating skin photoaging, wherein the α-ketobutyrate increases the expression of ZO-1 protein in mouse skin tissue.

[0010] The α-ketobutyrate is used in the preparation of skin care products and medicines for preventing and or treating skin photoaging, wherein the α-ketobutyrate increases the expression of Collagen I protein in mouse skin tissue.

[0011] The α-ketobutyrate is used in the preparation of skin care products and medicines for preventing and or treating skin photoaging, wherein the α-ketobutyrate reduces DNA damage in mouse skin tissue.

[0012] The α-ketobutyrate is used in the preparation of skin care products and medicines for preventing and or treating skin photoaging, wherein the α-ketobutyrate reduces the expression of aging markers in mouse skin tissue.

[0013] The above-mentioned α-ketobutyrate is used in the preparation of skin care products and medicines for preventing and or treating skin photoaging, wherein the aging markers include p53 and p21 proteins.

[0014] The above-mentioned α-ketobutyrate is used in the preparation of skin care products and medicines for preventing and or treating skin photoaging, wherein the dosage forms of the medicines include oily matrix ointments, emulsion matrix ointments and water-soluble matrix ointments.

[0015] The use of the above-mentioned α-ketobutyrate in the preparation of skin care products and medicines for preventing and or treating skin photoaging, wherein the dosage of the α-ketobutyrate is not less than 0.37 mg / cm 2 .

[0016] The present invention discloses an application of α-ketobutyrate in the preparation of skin care products and medicines for preventing and or treating photoaging of the skin. α-ketobutyrate is an endogenous metabolite in the human body. Experiments have shown that α-ketobutyrate treatment can significantly improve the skin manifestations of photoaging model mice, such as a large number of erythema, wrinkles, desquamation, inflammation, and thickening of the stratum corneum. α-ketobutyrate treatment can significantly increase the expression of E-Cadherin, ZO-1, and Collagen I in the skin tissue of photoaging model mice. α-ketobutyrate treatment can significantly reduce the expression of DNA damage and p53 and P21 aging markers in the skin tissue of photoaging model mice. At the same time, α-ketobutyric acid is a metabolite of the transsulfurization pathway in the human body, has strong pharmacological effects, few side effects, and avoids causing allergic reactions. Therefore, it provides a basis for the clinical treatment of skin photoaging and light damage. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 In chronic photoaging mice, α-ketobutyrate treatment (0.37 mg / cm 2 ) Skin pathological phenotype results at 7 weeks;

[0018] Figure 2 In chronic photoaging mice, α-ketobutyrate treatment (0.74 mg / cm 2 and smear 1.11mg / cm 2 ) Skin pathological phenotype results at 7 weeks;

[0019] Figure 3 In chronic photoaging mice, α-ketobutyrate treatment (0.74 mg / cm 2 and smear 1.11mg / cm 2 ) HE staining results of skin at 7 weeks;

[0020] Figure 4 A is a mouse model of chronic photoaging treated with α-ketobutyrate (applying 0.37 mg / cm 2 ) Western Blot detection of skin p21 protein at 7 weeks;

[0021] Figure 4 B is a picture of a mouse with chronic photoaging treated with α-ketobutyrate (applying 0.37 mg / cm 2 ) Immunohistochemical detection results of skin p21 protein at 7 weeks;

[0022] Figure 5 In chronic photoaging mice, α-ketobutyrate treatment (0.74 mg / cm 2 ) Western Blot analysis of skin E-Cadherin, ZO-1, and Collagen I proteins at 7 weeks;

[0023] Figure 6 In chronic photoaging mice, α-ketobutyrate treatment (0.74 mg / cm 2 ) Western Blot analysis of skin γH2Ax protein at 7 weeks;

[0024] Figure 7 In chronic photoaging mice, α-ketobutyrate treatment (0.74 mg / cm 2 ) Western Blot detection results of skin p53 and p21 proteins at 7 weeks. DETAILED DESCRIPTION

[0025] In order to enable those skilled in the art to better understand the technical solution of the present invention, the specific implementation methods thereof are described in detail below with reference to the accompanying drawings:

[0026] The present invention provides the use of α-ketobutyrate in the preparation of skin care products and medicines for preventing and or treating skin photoaging. The structural formula of α-ketobutyrate is:

[0027]

[0028] In the present invention, the molecular formula of α-ketobutyrate is CH3CH2COCOONa, and the molecular weight of α-ketobutyrate is 124.07. The present invention does not particularly limit the source of α-ketobutyrate, and any source of α-ketobutyrate known in the art can be used. In the present embodiment, the α-ketobutyrate was purchased from Shanghai MacLean Biochemical Technology Co., Ltd. under the product number S844402.

[0029] The present invention uses Balb / c mice to construct a skin photoaging mouse model using a solar simulator as the subject, and α-ketobutyrate is applied topically for 7 weeks. The results show that α-ketobutyrate treatment can significantly improve the skin symptoms of photoaging model mice, such as a large number of erythema, wrinkles, desquamation, inflammation, and thickening of the stratum corneum. α-ketobutyrate treatment can significantly increase the expression of E-Cadherin, ZO-1, and Collagen I in the skin tissue of photoaging model mice. α-ketobutyrate treatment can significantly reduce DNA damage and the expression of p53 and p21 aging markers in the skin tissue of photoaging model mice. Therefore, the present invention preferably provides the use of α-ketobutyrate in the preparation of skin care products and drugs for preventing and / or treating skin photoaging.

[0030] In the present invention, the preferred dosage form of the drug includes ointments including oily base ointments, emulsion base ointments and water-soluble base ointments. The dosage of the α-ketobutyrate is preferably not less than 0.37 mg / cm 2 , more preferably 0.74 to 1.11 mg / cm 2.

[0031] The application of the α-ketobutyrate provided by the present invention in the preparation of skin care products and medicines for preventing and / or treating skin photoaging is described in detail below with reference to the examples, but they should not be construed as limiting the scope of protection of the present invention.

[0032] Example 1

[0033] Establishment of a chronic photoaging mouse model

[0034] 1. Feeding Methods for Mice

[0035] 6-8 weeks old Balb / c female mice, SPF grade, weighing 20±2g, were purchased from the Department of Experimental Animals of Kunming Medical University (SCXK(Yunnan)K2020-0004). 6 mice / cage were kept in a 12-h light-dark cycle and temperature-controlled environment. The temperature was 26±2°C, the relative humidity was 55%±5%, and the animals had free access to food and water. The drinking water for the experimental animals was tap water that was packaged after autoclaving. All animal experiments in this example were carried out in accordance with the regulations for animal experiments. The Animal Ethics Committee of Kunming Medical University reviewed and approved the animal ethics of this study.

[0036] 2. Construction of a Skin Photoaging Mouse Model Using a Solar Simulator

[0037] Two days before the experiment, mice were anesthetized with 1.25% Avertin (Aibei Biotechnology) via intraperitoneal injection at a dose of 0.2 mL / 10 g. After anesthesia, the hair on the back of the mice was removed using an eyebrow razor. Gentle movements were performed to avoid damage to the skin. Depilatory creams were not used to prevent chemical agents from affecting the experimental results.

[0038] Photoirradiation: A Sigma solar simulator (Shanghai Sigma SUV1000) was used to perform photoaging ultraviolet irradiation. UVA and UVB filters were used to simulate the UV dose of solar radiation, with UVA accounting for 95% ± 3% and UVB accounting for 5% ± 3%. Exposure was performed five times per week for a total of nine weeks. The UV dose table is shown in Table 1: The initial dose was 1 MED (UVB approximately 0.07 J / cm 2 , UVA is about 1.37J / cm 2 ), with an irradiation time of 34 seconds per exposure. The dose was then increased by 0.5 MED per week, i.e., the second week's irradiation dose was 1.5 MED, the third week's irradiation dose was 2 MED, and so on until the model was complete. Before each irradiation, an ultraviolet radiation detection instrument was used to measure the energy of UVA and UVB to ensure that the ultraviolet radiation energy was constant during each exposure and reduce errors.

[0039] Table 1, UV radiation dose table:

[0040]

[0041] The photoaging phenotype indicators of the skin of photoaging model mice are a large number of erythema, loosening, roughness, inflammation and other phenotypes on the skin appearance.

[0042] Experiments have shown that α-ketobutyrate treatment can significantly improve the skin symptoms of photoaging model mice, such as numerous erythema, wrinkles, desquamation, inflammation, and thickening of the stratum corneum.

[0043] Example 2

[0044] Methods for the treatment of skin photoaging mouse model with α-ketobutyrate (α-KB) administration.

[0045] 1. Grouping method of control group and treatment group

[0046] The mice were divided into a normal mouse group, a photoaging mouse model group, and a drug-treated group (photoaging mice + α-KB), with 6 mice in each group and repeated 3 times. The drug-treated group was started after 2 weeks of modeling, and α-ketobutyrate was administered at a rate of 0.37 mg / cm 2 , 0.74mg / cm 2 and 1.11 mg / cm 2 The standard dosage of the drug was administered by applying it to the back once daily for 7 weeks. The normal mice and the photoaging model mice continued to be fed their respective diets. The mice were observed in real time during the experiment.

[0047] See the results Figure 1 α-ketobutyrate treatment was performed, applying 0.37 mg / cm 2 dose can improve the photoaging phenotype of mouse skin.

[0048] See the results Figure 2 α-ketobutyrate treatment was performed, applying 0.74 mg / cm 2 Dose and 1.11mg / cm 2 The dose can significantly improve the photoaging phenotype of mouse skin.

[0049] See the results Figure 3 α-ketobutyric acid treatment was performed, applying 0.74 mg / cm 2 Dose and 1.11mg / cm 2 The dose can improve the inflammatory condition of the mouse skin.

[0050] 2. Detection of Biochemical Indices in Mouse Skin Tissue

[0051] Mice in each group were sacrificed, and skin tissues were isolated. Skin protein samples were extracted and assayed for protein concentration using the BCA assay. Proteins were separated by SDS-PAGE electrophoresis. Immunoblotting was performed for E-cadherin, ZO-1, Collagen I, γH2Ax, p53, and p21. GAPDH was used as an internal control protein.

[0052] See also Figure 4 , α-ketobutyrate treatment was performed, applying 0.37mg / cm 2 The dose can significantly reduce the expression of P21 in the skin tissue of photoaged mice.

[0053] See also Figure 5 , α-ketobutyrate treatment was performed, applying 0.74mg / cm 2 Dose and 1.11mg / cm 2 The dose can significantly increase the expression of E-Cadherin, ZO-1 and Collagen I in the skin tissue of photoaged mice.

[0054] See also Figure 6 , α-ketobutyrate treatment was performed, applying 0.74mg / cm 2 Dose and 1.11mg / cm 2 The dose can significantly reduce DNA damage (γH2Ax) in the skin tissue of photoaging model mice.

[0055] See also Figure 7 , α-ketobutyrate treatment was performed, applying 0.74mg / cm 2 Dose and 1.11mg / cm 2 The dose can significantly reduce the expression of p53 and p21 aging markers in the skin tissue of photoaging model mice.

[0056] α-Ketobutyrate is used in the preparation of skin care products and drugs for preventing and / or treating skin photoaging by improving erythema, wrinkles, desquamation, inflammation, and stratum corneum thickening in mouse skin. α-Ketobutyrate is used in the preparation of skin care products and drugs for preventing and / or treating skin photoaging by increasing the expression of E-Cadherin protein, ZO-1 protein, and Collagen I protein in mouse skin tissue. α-Ketobutyrate is used in the preparation of skin care products and drugs for preventing and / or treating skin photoaging by reducing DNA damage and the expression of p53 and P21 aging markers in mouse skin tissue. The dosage forms of the drug include ointments, including oily base ointments, emulsion base ointments, and water-soluble base ointments. α-Ketobutyrate is used in the development of skin care products that assist in the prevention and treatment of photoaging.

[0057] In summary, the use of α-ketobutyrate of the present invention in the preparation of skin care products and medicines for preventing and or treating skin photoaging. α-ketobutyrate is a metabolite of the transsulfurization pathway in the human body, has strong pharmacological effects, few side effects, and avoids causing allergic reactions, providing a basis for the clinical treatment of skin photoaging and light damage.

[0058] Those skilled in the art should recognize that the above embodiments are merely intended to illustrate the present invention and are not intended to limit the present invention. As long as they are within the spirit of the present invention, any changes or modifications to the above embodiments will fall within the scope of the claims of the present invention.

Claims

1. Use of α-ketobutyrate in the preparation of skin care products and medicines for preventing and or treating skin photoaging, characterized in that: The structural formula of the α-ketobutyrate is:

2. The use of α-ketobutyrate according to claim 1 in the preparation of skin care products and medicines for preventing and or treating skin photoaging, characterized in that: The α-ketobutyrate improves erythema, wrinkles, desquamation, inflammation and stratum corneum thickening of mouse skin.

3. The use of α-ketobutyrate according to claim 1 in the preparation of skin care products and medicines for preventing and or treating skin photoaging, characterized in that: The α-ketobutyrate increases the expression of E-Cadherin protein in mouse skin tissue.

4. The use of α-ketobutyrate according to claim 1 in the preparation of skin care products and medicines for preventing and or treating skin photoaging, characterized in that: The α-ketobutyrate increases the expression of ZO-1 protein in mouse skin tissue.

5. The use of α-ketobutyrate according to claim 1 in the preparation of skin care products and medicines for preventing and or treating skin photoaging, characterized in that: The α-ketobutyrate increases the expression of Collagen I protein in mouse skin tissue.

6. The use of α-ketobutyrate according to claim 1 in the preparation of skin care products and medicines for preventing and or treating skin photoaging, characterized in that: The α-ketobutyrate reduces DNA damage in mouse skin tissue.

7. The use of α-ketobutyrate according to claim 1 in the preparation of skin care products and medicines for preventing and or treating skin photoaging, characterized in that: The α-ketobutyrate reduces the expression of aging markers in mouse skin tissue.

8. The use of α-ketobutyrate according to claim 7 in the preparation of skin care products and medicines for preventing and or treating skin photoaging, characterized in that: The aging markers include p53 and p21 proteins.

9. The use of α-ketobutyrate according to claim 1 in the preparation of skin care products and medicines for preventing and or treating skin photoaging, characterized in that: The dosage forms of the medicine include oily base ointment, emulsion base ointment and water-soluble base ointment.

10. The use of α-ketobutyrate according to claim 1 in preparing skin care products and medicines for preventing and or treating skin photoaging, characterized in that: The dosage of the α-ketobutyrate is not less than 0.37 mg / cm 2 .

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