Use of IdeS

Abstract

The application provides an IdeS application for preparing a product for coping with skin photoaging, the product including preparation of a skin care product and a medicine for preventing and / or treating skin photoaging, a dosage form of the medicine including an ointment, and the skin care product is applied to assist in preventing and treating skin photoaging. The IdeS treatment can significantly improve a large number of erythema, wrinkles, desquamation, inflammation and keratin layer thickening and other performances presented by the skin of a photoaging model mouse, has been developed as a new type of biological treatment medicine, has strong pharmacological effect, small side effect, and can avoid causing an allergic reaction, and therefore provides a basis for clinically treating skin photoaging and photo-damage.

Description

Technical Field

[0001] This invention belongs to the field of skin disease biomedical technology, specifically involving the application of an IdeS. Background Technology

[0002] The skin is the largest organ in the human body and the first line of defense against external physical, chemical, and biological stimuli. Photoaging, also known as extrinsic aging, refers to premature aging caused by prolonged and repeated exposure to sunlight, especially ultraviolet radiation. Unlike natural aging (intrinsic aging), photoaging mainly manifests as roughness, thickening, dryness, sagging, deepened and coarser wrinkles, uneven pigmentation (such as age spots), and telangiectasia on exposed areas of the skin (such as the face, neck, and back of the hands). In severe cases, it can even induce skin tumors such as actinic keratosis, squamous cell carcinoma, and basal cell carcinoma. Epidemiological surveys show that over 80% of facial skin aging can be attributed to sun exposure. Photoaging has become a global public health problem, not only affecting patients' appearance and psychology but also significantly increasing the risk of skin cancer.

[0003] Currently, clinical methods for preventing or treating photoaging of the skin are mainly divided into three categories. The first category is physical / chemical protection, such as using sunscreen and umbrellas to reduce ultraviolet radiation exposure. However, sun protection measures can only prevent or delay the onset of photoaging; damage caused by photoaging that has already occurred is almost irreversible. The second category is drug intervention, among which retinoids (such as all-trans retinoic acid and tazarotene) are currently the only topical medications approved by the US FDA for treating photoaged skin. These drugs improve wrinkles and pigmentation by regulating epidermal differentiation, inhibiting matrix metalloproteinase (MMP) activity, and promoting collagen synthesis. However, retinoids have strong skin irritation; almost all patients experience adverse reactions such as erythema, desquamation, burning sensation, and increased photosensitivity in the early stages of treatment, leading to poor patient compliance. Furthermore, oral retinoids may cause systemic side effects such as teratogenicity, dyslipidemia, and liver damage, limiting their long-term use. The third category is physical / phototherapy, such as fractional laser, intense pulsed light, radiofrequency ablation, and photodynamic therapy. These methods stimulate dermal collagen remodeling through the "limited damage and reconstruction" mechanism, which can improve wrinkles and skin texture to some extent. However, the treatment process is painful and expensive, and there are risks of postoperative erythema, edema, pigmentation, and even scar formation. Multiple treatments and strict postoperative care are usually required.

[0004] While the aforementioned existing technologies can alleviate the clinical manifestations of photoaging to some extent, their limitations are obvious. Furthermore, these treatments fail to simultaneously address multiple core pathological aspects of photoaging at the molecular level: UV-induced DNA damage, collagen degradation due to MMP-1 overexpression, downregulation of epidermal barrier proteins (E-Cadherin, ZO-1), and the accumulation of cellular aging markers (elevated p21, decreased Lamin B1). Therefore, there is a need to design an application for IdeS. Summary of the Invention

[0005] To overcome the shortcomings of existing technologies, an application of IdeS is provided.

[0006] To achieve the above objectives, the present invention provides the following technical solution:

[0007] An application of IdeS, wherein the IdeS is used to prepare products that address photoaging of the skin.

[0008] The amino acid sequence of the IdeS is shown in SEQ ID NO.1.

[0009] The products include skin care products and pharmaceuticals for the prevention and / or treatment of photoaging of the skin.

[0010] The dosage form of the drug includes ointment.

[0011] The ointment includes oil-based ointments, emulsion-based ointments, and water-soluble base ointments.

[0012] The skincare products mentioned are used to help prevent and treat photoaging of the skin.

[0013] The dosage of IdeS is not less than 0.12 mg / kg.

[0014] The route of administration is subcutaneous injection.

[0015] Compared with the prior art, the advantages and beneficial effects of the present invention are as follows:

[0016] This invention provides the application of IdeS, a protein with the structure shown in Formula I, in the preparation of skincare products and drugs for the prevention and / or treatment of photoaging. Experiments show that IdeS treatment significantly improves the extensive erythema, wrinkles, desquamation, inflammation, and thickening of the stratum corneum observed in photoaged mouse skin. IdeS treatment significantly increases the expression of E-Cadherin, ZO-1, and Collagen I in the skin tissue of photoaged mice. IdeS treatment significantly reduces the expression of DNA damage, MMP-1, and the aging marker p21 in the skin tissue of photoaged mouse models, and significantly increases the expression of the aging marker Lamin B1. Furthermore, IdeS has been developed as a novel biological therapeutic drug with strong pharmacological effects, few side effects, and avoidance of allergic reactions, thus providing a basis for the clinical treatment of photoaging and photodamage. Attached Figure Description

[0017] Figure 1 The results of skin pathology in chronically photoaged mice treated with IdeS (0.12 mg / kg subcutaneously) for 7 weeks are shown.

[0018] Figure 2 HE staining results of skin treated with IdeS (0.12 mg / kg subcutaneously) for 7 weeks in chronically photoaged mice;

[0019] Figure 3 To detect the immunofluorescence of E-Cadherin and ZO-1 proteins in the skin of chronically photoaged mice after IdeS treatment (0.12 mg / kg subcutaneously) for 7 weeks;

[0020] Figure 4 Immunohistochemical assay of skin MMP-1 and Collagen I proteins in chronically photoaged mice treated with IdeS (subcutaneous injection of 0.12 mg / kg) for 7 weeks.

[0021] Figure 5 To detect the immunofluorescence of skin DNA damage markers (γ-H2AX protein) in chronically photoaged mice after 7 weeks of IdeS treatment (subcutaneous injection of 0.12 mg / kg);

[0022] Figure 6 Immunohistochemical detection of skin p21 protein in chronically photoaged mice treated with IdeS (0.12 mg / kg subcutaneously) for 7 weeks;

[0023] Figure 7 Immunofluorescence assay of skin Lamin B1 protein in chronically photoaged mice treated with IdeS (0.12 mg / kg subcutaneously) for 7 weeks;

[0024] Figure 8This is a schematic diagram of the protein structure of IdeS described in this application. Detailed Implementation

[0025] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0026] The technical solution of this application is as follows:

[0027] An application of an IdeS, wherein the IdeS is used to prepare products addressing skin photoaging. This invention provides a protein structure having, as... Figure 8 The IdeS shown is used in the preparation of skin care products and medicines for the prevention and / or treatment of photoaging of the skin.

[0028] The amino acid sequence of the IdeS is shown in SEQ ID NO.1.

[0029] The SEQ ID NO.1 is:

[0030] DSFSANQEIRYSEVTPYHVTSVWTKGVTPPAKFTQGEDVFHAPYVANQGWYDITKTFNGKDDLLCGAATAGNMLHWWFDQNKEKIEAYLKKHPDKQKIMFGDQELLDVRKVINTKGDQTNSELFNYFRDKAFPGLSARRIGVMPDLVLDMFINGYY LNVYKTQTTDVNRTYQEKDRRGGIFDAVFTRGDQSKLLTSRHDFKEKNLKEISDLIKKELTEGKALGLSHTYANVRINHVINLWGADFDSNGNLKAIYVTDSDSNASIGMKKYFVGVNSAGKVAISAKEIKEDNIGAQVLGLFTSTGQDSWNQTN.

[0031] The products include skin care products and pharmaceuticals for the prevention and / or treatment of photoaging of the skin.

[0032] The dosage form of the drug includes ointment.

[0033] The ointment includes oil-based ointments, emulsion-based ointments, and water-soluble base ointments.

[0034] The skincare products mentioned are used to help prevent and treat photoaging of the skin.

[0035] The dosage of IdeS is not less than 0.12 mg / kg. The route of administration is subcutaneous injection.

[0036] In this invention, IdeS is a highly specific IgG endopeptidase evolved from Streptococcus pyogenes. This invention does not have any particular restrictions on the source of IdeS; any source of IdeS known in the art can be used. In this embodiment, the IdeS was purchased from MedChemExpress (MCE China), catalog number HY-P74867.

[0037] This invention uses Balb / c mice to construct a photoaging mouse model of skin using a sunlight simulator, and intervenes with IdeS drug administration. Results show that IdeS intervention significantly improves the pathological phenotypes of photoaging in the mouse skin, including extensive erythema, wrinkles, desquamation, inflammation, and thickened stratum corneum. IdeS intervention significantly increases the expression of E-Cadherin, ZO-1, and Collagen I in the skin tissue of the photoaging mouse model. IdeS intervention significantly decreases the expression of MMP-1, DNA damage, and the aging marker p21 in the skin tissue of the photoaging mouse model. IdeS intervention significantly increases the expression of the aging marker Lamin B1 in the skin tissue of the photoaging mouse model. Furthermore, IdeS is a highly specific IgG degrading enzyme, used for the intervention of antibody-mediated immune diseases, exhibiting strong pharmacological effects and few side effects, thus providing a basis for the clinical development of skin care products and drugs for treating photoaging.

[0038] This invention provides the application of IdeS (Immunoglubulin G-degrading enzyme of Streptococcus pyogenes), a specific protease derived from Streptococcus pyogenes, in the preparation of skin care products and drugs for the prevention and / or treatment of photoaging. It can eliminate autoimmune reactions, has fewer side effects, and targets mainly MMP-1, DNA damage, and aging markers. IdeS shows great promise in the clinical treatment of photoaging skin diseases.

[0039] The present invention will be described in detail below through examples and comparative examples, but the scope of protection of the present invention is not limited to these examples. Unless otherwise specified, the chemical reagents and raw materials used in the following examples and comparative examples are all conventional commercially available products.

[0040] Example 1

[0041] Establishment of a mouse model of chronic photoaging

[0042] 1. Feeding methods for mice

[0043] Six- to eight-week-old female Balb / c mice, specific pathogen free (SPF) grade, weighing 20±2 g, were purchased from the Experimental Animal Center of Yunnan University (SCXK(Dian) K2021-0001). The mice were housed at 6 mice per cage in a temperature-controlled environment with a 12 h light-dark cycle. The temperature was 26±2 °C, the relative humidity was 55%±5%, and the mice had free access to food and drinking water. The drinking water for experimental animals was tap water that was autoclaved and then aliquoted. All animal experiments in this example were carried out in accordance with the regulations for animal experiments. The animal ethics review of this study was reviewed and approved by the Animal Ethics Committee of Kunming Medical University.

[0044] 2. Construction of a mouse model of skin photoaging using a solar simulator

[0045] Two days before the experiment, mice were anesthetized by intraperitoneal injection of 1.25% Avertin (Aibei Biotech) at a dose of 0.2 ml / 10 g. After anesthesia, the hair on the back of the mice was removed completely with an eyebrow razor. The hair removal should be performed gently to avoid artificial damage to the skin. Depilatory cream is not used for hair removal to avoid the influence of chemical reagents on the experimental results.

[0046] Irradiation treatment: The Sigma solar simulator (Shanghai Sigma SUV1000) was used for photoaging ultraviolet irradiation: UVA+UVB filters were used to simulate the dose of ultraviolet in solar radiation, that is, UVA accounts for 95%±3% and UVB accounts for 5%±3%. Irradiation was performed 5 times a week for a total of 9 weeks: The initial irradiation dose was 1 MED (approximately 0.07 J / cm2 for UVB and approximately 1.37 J / cm2 for UVA), and the irradiation time was 34 s per time; thereafter, the dose was increased by 0.5 MED every week, that is, the irradiation dose was 1.5 MED in the second week, 2 MED in the third week, and so on until the end of modeling. Before each irradiation, an ultraviolet radiation detector was used to detect the energy of UVA and UVB, so as to ensure that the ultraviolet radiation energy was constant in each irradiation and reduce errors.

[0047] Ultraviolet irradiation dose

[0048] WeekSingle irradiation durationSingle irradiation doseCumulative irradiation dose

[0049] Week 134 sUVB: 0.07 J / cm2+UVA: 1.37 J / cm2UVB: 0.35 J / cm2+UVA: 6.85 J / cm2

[0050] Week 250 sUVB: 0.105 J / cm2+UVA: 2.055 J / cm2UVB: 0.875 J / cm2+UVA: 17.125 J / cm2

[0051] Week 3, 67 sUVB: 0.14 J / cm2 +UVA: 2.74 J / cm2;UVB: 1.575 J / cm2 +UVA: 30.825 J / cm2

[0052] Week 4: 84 sUVB: 0.175 J / cm² + UVA: 3.425 J / cm² UVB: 2.45 J / cm² + UVA: 47.95 J / cm²

[0053] Week 5: 100 sUVB: 0.21 J / cm² + UVA: 4.11 J / cm²; UVB: 3.5 J / cm² + UVA: 68.5 J / cm²

[0054] Week 6 sUVB: 0.245 J / cm² + UVA: 4.795 J / cm² UVB: 4.725 J / cm² + UVA: 92.475 J / cm²

[0055] Week 7: 134 sUVB: 0.28 J / cm² + UVA: 5.48 J / cm²; UVB: 6.125 J / cm² + UVA: 119.875 J / cm²

[0056] Week 8: 150 sUVB: 0.315 J / cm² + UVA: 6.165 J / cm²; UVB: 7.7 J / cm² + UVA: 150.7 J / cm²

[0057] Week 9: 167 sUVB: 0.35 J / cm² + UVA: 6.85 J / cm² UVB: 9.45 J / cm² + UVA: 184.95 J / cm²

[0058] The photoaging phenotype of mice skin showed numerous erythema, loosening, roughness, and inflammation.

[0059] Experiments have shown that IdeS treatment can significantly improve the skin manifestations of photoaging model mice, such as extensive erythema, wrinkles, desquamation, inflammation, and thickening of the stratum corneum.

[0060] Example 2

[0061] A method for treating a mouse model of skin photoaging with IdeS drug delivery.

[0062] 1. Grouping method for control group and treatment group

[0063] The mice were divided into three groups: a normal mouse group, a photo-aged mouse model group, and a drug-treated group (photo-aged mice + IdeS), with six mice in each group and three replicates per group. In the drug-treated group, administration began two weeks after the onset of photoaging. IdeS was administered subcutaneously at a standard rate of 0.12 mg / kg once a week for seven weeks. The normal mouse group and the photo-aged mouse model group continued to be fed according to their respective diets. The mice were monitored in real-time throughout the experiment.

[0064] See results Figure 1 Subcutaneous injection of 0.12 mg / kg of IdeS treatment significantly improved photoaging phenotypes in mice, such as erythema, wrinkles, and desquamation.

[0065] See results Figure 2 Subcutaneous injection of 0.12 mg / kg of IdeS treatment improved the inflammatory condition of the skin in mice.

[0066] 2. Detection of biochemical indicators in mouse skin tissue

[0067] Mice in each group were sacrificed, and skin tissue was isolated. After fixation in paraformaldehyde, the tissue was embedded in paraffin and sectioned. Antibodies were used as probes to locate and qualitatively analyze specific antigens within the tissue. Finally, immunofluorescence / histochemical experiments were performed on E-Cadherin, ZO-1, Collagen I, MMP-1, γ-H2AX, Lamin B1, and p21.

[0068] like Figure 3 As shown, IdeS treatment, with subcutaneous injection of 0.12 mg / kg, significantly increased the expression of E-Cadherin and ZO-1 in the skin tissue of photoaged mice, stabilizing the skin barrier.

[0069] like Figure 4 As shown, IdeS treatment, with subcutaneous injection of 0.12 mg / kg, reduced the expression of MMP-1 in the skin tissue of photoaged mice and significantly inhibited the degradation of Collagen I, thus reducing the formation of wrinkles.

[0070] like Figure 5 As shown, IdeS treatment, with subcutaneous injection of 0.12 mg / kg, significantly reduced DNA damage (γ-H2AX) in the skin tissue of photoaged mice.

[0071] like Figure 6 As shown, IdeS treatment, with subcutaneous injection of 0.12 mg / kg, significantly reduced the expression of p21 in the skin tissue of photoaged mice.

[0072] like Figure 7As shown, IdeS treatment, with subcutaneous injection of 0.12 mg / kg, significantly increased the expression of Lamin B1 in the skin tissue of photoaged mice.

[0073] IdeS is used in the preparation of skincare products and drugs for the prevention and / or treatment of photoaging by improving erythema, wrinkles, desquamation, inflammation, and thickening of the stratum corneum in mouse skin. IdeS is also used in the preparation of skincare products and drugs for the prevention and / or treatment of photoaging by increasing the expression of E-Cadherin, ZO-1, and Collagen I proteins in mouse skin tissue. Furthermore, IdeS is used in the preparation of skincare products and drugs for the prevention and / or treatment of photoaging by decreasing the expression of p21, a DNA damage and aging marker, in mouse skin tissue. IdeS is also used in the preparation of skincare products and drugs for the prevention and / or treatment of photoaging by increasing the expression of Lamin B1, an aging marker, in mouse skin tissue. The drug dosage forms include ointments, including oil-based ointments, emulsion-based ointments, and water-soluble ointments. IdeS is used in the research and development of skincare products that aid in the prevention and treatment of photoaging.

[0074] In summary, the application of IdeS in the preparation of skin care products and drugs for the prevention and / or treatment of photoaging of the skin, IdeS is a metabolite of the transsulfurization pathway in the human body with strong pharmacological effects, few side effects, and avoidance of allergic reactions, providing a basis for the clinical treatment of photoaging and photodamage of the skin.

[0075] The above description represents the preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. An application of IdeS, characterized in that, The IdeS is used to prepare products for the prevention and / or treatment of skin photoaging; the amino acid sequence of the IdeS is shown in SEQ ID NO.

1.

2. An application of IdeS according to claim 1, characterized in that, The products include skincare products and pharmaceuticals.

3. An application of IdeS according to claim 2, characterized in that, The dosage form of the drug includes ointment.

4. An application of IdeS according to claim 3, characterized in that, The ointment includes oil-based ointments, emulsion-based ointments, and water-soluble base ointments.

5. An application of IdeS according to claim 2, characterized in that, The skincare products mentioned are used to help prevent and treat photoaging of the skin.

6. An application of IdeS according to claim 2, characterized in that, The dosage of IdeS is not less than 0.12 mg / kg.

7. An application of IdeS according to claim 2, characterized in that, The route of administration is subcutaneous injection.

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