Application of apocynin in preparation of medicine for preventing and / or treating depigmentation dermatosis

Abstract

The invention provides application of apocynin in preparation of a medicine for preventing and / or treating depigmentation skin diseases, and belongs to the technical field of skin disease medicine manufacturing. The invention discloses application of apocynin or an apocynin derivative in preparation of a medicine for preventing and / or treating depigmentation skin diseases, especially vitiligo. The medicine prepared from the apocynin can effectively recover leucoderma model mouse skin pigments, effectively reduce expression of model mouse skin inflammatory factors and reduce infiltration of CD8 + T cells. Therefore, the apocynin or the apocynin derivative achieves the purpose of treating depigmentation skin diseases by relieving skin inflammation, protecting melanocytes from being oxidized and damaged and promoting melanogenesis. The invention not only expands the medical application of apocynin, but also provides a new way for clinical treatment of depigmentation dermatosis.

Description

Technical Field

[0001] This invention belongs to the field of dermatological drug manufacturing technology, specifically relating to the application of Apocynum venetum in the preparation of drugs for the prevention and / or treatment of depigmenting skin diseases. Background Technology

[0002] Depigmentative skin diseases are a collective term for a group of diseases caused by abnormal function (reduction or loss) or a decrease in the number of melanocytes in the skin, resulting in white patches on the skin. All types of diseases revolve around "disorders of melanin synthesis." Melanin is produced by melanocytes in the skin; once these cells are damaged, their function is impaired, or they cannot migrate normally, the skin turns white due to a lack of pigment. Causes include genetic factors (such as albinism and piebaldism), autoimmune diseases (such as vitiligo), inflammation or trauma (such as post-inflammatory hypopigmentation), and environmental stimuli (such as excessive sun exposure and chemical contact). Vitiligo is a skin disease caused by chronic inflammation, characterized by progressive depigmentation of the skin and mucous membranes. The core pathology of vitiligo is the absence or dysfunction of functional melanocytes in the skin, leading to localized or generalized white patches on the skin.

[0003] Existing treatments for depigmenting skin diseases mainly include glucocorticoids, calcineurin inhibitors, and narrowband UVB phototherapy. Although these methods have some efficacy, they also have problems such as significant side effects and incomplete treatment. Summary of the Invention

[0004] This invention provides the use of Apocynum venetum in the preparation of medicaments for the prevention and / or treatment of depigmenting skin diseases.

[0005] This invention provides the use of apocynum or apocynum derivatives in the preparation of medicaments for the prevention and / or treatment of depigmenting skin diseases.

[0006] Preferably, the depigmenting skin disease includes vitiligo.

[0007] Preferably, the treatment of depigmented skin diseases includes at least one of the following: promoting the increase of melanocytes in the skin, promoting melanin synthesis, inhibiting melanocyte apoptosis, and protecting melanocytes from oxidative damage.

[0008] Preferably, the dosage form of the drug includes at least one of the following external dosage forms: cream, ointment, gel, lotion, liniment, and lotion.

[0009] Preferably, the drug further includes pharmaceutically acceptable excipients.

[0010] Preferably, the apocynum derivative includes apocynum salt derivatives.

[0011] Preferably, the mass percentage of apocynum or apocynum derivative in the drug is 0.1% to 10%.

[0012] This invention provides a cream for treating pigmented skin diseases, comprising the following components in parts by weight: 0.5-1.5 parts of Apocynum venetum, 2-4 parts of glyceryl mono- and di-stearyl esters, 4-6 parts of fatty alcohol, 3-5 parts of liquid paraffin, 0.08-0.12 parts of ethylparaben, 1.8-2.2 parts of polyoxyethylene (21) stearyl ether, 1.8-2.2 parts of polyoxyethylene (2) stearyl ether, 1.8-2.2 parts of polyglycerol oleate, 55-60 parts of ultrapure water, 7-9 parts of glycerol, and 13-17 parts of diethylene glycol monoethyl ether.

[0013] Preferably, the components include the following parts by weight: 1 part of Apocynum venetum, 3 parts of glyceryl mono- and di-stearyl esters, 5 parts of fatty alcohol, 4 parts of liquid paraffin, 0.1 parts of ethylparaben, 2 parts of polyoxyethylene (21) stearyl alcohol ether, 2 parts of polyoxyethylene (2) stearyl alcohol ether, 2 parts of polyglycerol oleate, 58 parts of ultrapure water, 8 parts of glycerol, and 15 parts of diethylene glycol monoethyl ether.

[0014] Preferably, the fatty alcohol includes hexadecyl alcohol and / or octadecyl alcohol.

[0015] This invention provides the application of Apocynum venetum or its derivatives in the preparation of drugs for the prevention and / or treatment of depigmenting skin diseases. The Apocynum venetum or its derivatives have the effect of restoring the number of melanocytes in the skin, providing a new approach for the treatment of depigmenting skin diseases whose pathological core is the loss or dysfunction of functional melanocytes in the skin. Experimental results of this invention show that the drug prepared from Apocynum venetum can effectively restore skin pigmentation in vitiligo model mice, thus having a therapeutic effect on vitiligo; simultaneously, Apocynum venetum can effectively reduce the expression of inflammatory factors (IFN-γ, TNF-α, IL-1β, IL-6) in the skin of model mice, and also reduce CD8... + T-cell infiltration and melanocyte-specific marker detection results indicate a significant increasing trend in melanocytes. Therefore, Apocynum venetum or its derivatives can achieve the therapeutic goal of treating depigmenting skin diseases by reducing skin inflammation, protecting melanocytes from oxidative damage, and promoting melanin production. Attached Figure Description

[0016] Figure 1 Images of the back skin of mice in the model group and treatment group at different time points; Figure 2 The results of serum inflammatory factor detection in mice in the model group and treatment group are shown below; where A is the result of plasma IFN-γ detection, B is the result of plasma TNF-α detection, C is the result of plasma IL-1β detection, and D is the result of plasma IL-6 detection; Note: ns indicates no significant difference between the two groups. Indicates a significant difference between the two groups. P <0.05; P Indicates a significant difference between the two groups. P <0.0001; Figure 3 HE and immunohistochemical staining results of mouse tissues from the model group and treatment group; scale bar is 100 μM. Figure 4 Masson staining results for model and treated mouse tissues; scale bar is 100 μM. Detailed Implementation

[0017] This invention provides the use of apocynum or apocynum derivatives in the preparation of medicaments for the prevention and / or treatment of depigmenting skin diseases.

[0018] In this invention, the molecular formula of the apocynum venetum is C9H10O3, the molecular weight is 166.17, and the structural formula is shown in Formula I. In this embodiment of the invention, the apocynum venetum was purchased from Nantong Feiyu Biotechnology Co., Ltd., No: FY26199; Lot: DGA133.

[0019] Formula I In this invention, the apocynum derivative preferably includes apocynum salt derivatives, such as inorganic acid salts and organic acid salts. The inorganic acid salts include hydrochloride, sulfate, and nitrate formed by the reaction of apocynum with hydrochloric acid, sulfate, and nitrate. The organic acid salts include acetate, maleate, tartrate, and citrate formed by the reaction of apocynum with acetic acid, maleic acid, tartaric acid, and citric acid.

[0020] In this invention, the depigmenting skin disease preferably includes vitiligo. Vitiligo is a skin disease caused by chronic inflammation, characterized by depigmentation, with the loss or dysfunction of functional melanocytes as the main pathological mechanism. Because the activity of NADPH oxidase (NOX) is significantly upregulated in the skin lesions of vitiligo patients, leading to a large production of ROS, high levels of ROS have direct cytotoxicity to melanocytes, inducing their apoptosis; simultaneously, ROS also acts as a signaling molecule to activate dendritic cells, presenting melanocyte-associated antigens, thereby activating cytotoxic T cells (CD8+). + NOX attacks and destroys melanocytes; in addition, ROS can inhibit the activity of key enzymes in melanin synthesis (such as tyrosinase) and impair skin barrier function. Therefore, inhibiting NOX activity and blocking ROS synthesis are important pathways to suppress melanocyte destruction and synthesis disorders in vitiligo.

[0021] In this invention, the treatment of depigmented skin diseases preferably includes at least one of the following: promoting the increase of melanocytes in the skin, promoting melanin synthesis, inhibiting melanocyte apoptosis, and protecting melanocytes from oxidative damage.

[0022] In one embodiment of the present invention, a drug with Apocynum venetum as the active ingredient was applied topically to a hydroquinone-induced vitiligo model mouse. The results showed that repigmentation of the back skin was clearly observed in the Apocynum venetum treatment group, indicating that Apocynum venetum has an effective therapeutic effect on vitiligo. Melan-A staining and SOX10 staining were used to detect the expression of melanocyte-specific markers. The results showed that the Apocynum venetum treatment group could reverse the reduction of melanocyte-specific markers in the model group mice. The increase in melanocyte-specific markers in the Apocynum venetum treatment group indicates that Apocynum venetum can increase the number of melanocytes in the skin and promote melanin synthesis.

[0023] In this invention, the described Apocynum venetum glycoside has the activity of inhibiting NADPH oxidase (NOX). NADPH oxidase initiates an oxidative stress cascade by reducing molecular oxygen to produce ROS, and is one of the main enzyme systems that produce ROS in vivo. ROS can not only induce melanocyte apoptosis, but also attack melanocytes and inhibit the activity of key enzymes in melanin synthesis (such as tyrosinase), thus hindering melanin synthesis and damaging the skin barrier function. Therefore, based on the NOX-inhibiting activity of the described Apocynum venetum glycoside, the production of ROS can be effectively inhibited, thereby inhibiting melanocyte apoptosis, protecting melanocytes from oxidative damage, and promoting melanin synthesis.

[0024] In this invention, since depigmenting skin diseases include the formation of inflammatory inducing factors, and vitiligo is a skin disease caused by chronic inflammation, inhibiting the skin inflammatory response is also an important way to prevent or treat depigmenting skin diseases or vitiligo. In one embodiment of this invention, ELISA detection of inflammatory factor expression in skin tissue showed that, compared with the model group, the expression of inflammatory factors IFN-γ, TNF-α, IL-1β, and IL-6 in the serum of mice treated with Apocynum venetum significantly decreased, indicating that Apocynum venetum has the effect of reducing skin inflammatory response. This suggests that Apocynum venetum can also treat vitiligo by inhibiting skin inflammatory response.

[0025] In this invention, the dosage form of the drug preferably includes at least one of the following external dosage forms: cream, ointment, gel, emulsion, liniment, and lotion. The drug preferably also includes pharmaceutically acceptable excipients. Pharmaceutically acceptable carriers include diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorbents, lubricants, etc., conventional in the pharmaceutical field. "Pharmaceuticalally acceptable" refers to ligands, materials, compositions, and / or dosage forms that are suitable for administration to patients within the bounds of reasonable medical judgment and that are proportionate to a reasonable benefit / risk ratio. The drugs of all the above dosage forms can be prepared according to conventional methods in the pharmaceutical field.

[0026] The mass percentage of apocynum or apocynum derivative in the drug is preferably 0.1-10%, but can be 0.5-5% or 1-3%.

[0027] This invention provides a cream for treating pigmented skin diseases, comprising the following components in parts by weight: 0.5-1.5 parts of Apocynum venetum, 2-4 parts of glyceryl mono- and di-stearyl esters, 4-6 parts of fatty alcohol, 3-5 parts of liquid paraffin, 0.08-0.12 parts of ethylparaben, 1.8-2.2 parts of polyoxyethylene (21) stearyl ether, 1.8-2.2 parts of polyoxyethylene (2) stearyl ether, 1.8-2.2 parts of polyglycerol oleate, 55-60 parts of ultrapure water, 7-9 parts of glycerol, and 13-17 parts of diethylene glycol monoethyl ether.

[0028] In this invention, the cream preferably comprises the following components in parts by weight: 1 part Apocynum venetum, 3 parts glyceryl mono- and di-stearyl esters, 5 parts fatty alcohol, 4 parts liquid paraffin, 0.1 part ethylparaben, 2 parts polyoxyethylene (21) stearyl ether, 2 parts polyoxyethylene (2) stearyl ether, 2 parts polyglycerol oleate, 58 parts ultrapure water, 8 parts glycerol, and 15 parts diethylene glycol monoethyl ether. The fatty alcohol preferably includes hexadecyl alcohol and / or octadecyl alcohol.

[0029] In this invention, the method for preparing the cream preferably includes the following steps: Component A is obtained by mixing glyceryl monostearate and glyceryl distearate, fatty alcohol, liquid paraffin, ethylparaben, polyoxyethylene (21) stearyl alcohol ether, polyoxyethylene (2) stearyl alcohol ether, and polyglyceryl oleate. Ultrapure water and glycerol were mixed to obtain component B; Diethylene glycol monoethyl ether and Apocynum venetum were mixed and dissolved to obtain component C; The components A and B are mixed and homogenized under heating conditions to obtain a mixture; The mixture and component C are mixed and homogenized to obtain a paste.

[0030] In this invention, the preferred heating and dissolving temperature is 80-90°C, and can be 85°C. The preferred mixing time is 2-4 minutes, and can be 3 minutes. The homogenization of the mixture with component C is preferably carried out when the mixture is cooled to below 60°C. The ointment prepared by the above method has a uniform texture, good stability, and can maintain the efficacy of Apocynum venetum for a long time. It can also improve the adhesion of the ointment to the skin and improve the drug utilization rate of Apocynum venetum.

[0031] The following detailed description, in conjunction with embodiments, illustrates the application of the Apocynum venetum provided by the present invention in the preparation of medicaments for the prevention and / or treatment of pigmentary depigmentation skin diseases. However, these descriptions should not be construed as limiting the scope of protection of the present invention.

[0032] In this embodiment, the ELISA kits were purchased from Shanghai Weiao Biotechnology Co., Ltd.: Mouse IFN-γ ELISA kit (High Sensitivity), catalog number EM30253M; Mouse TNF-α ELISA kit, catalog number EM30536M; and Mouse IL-6 ELISA kit, catalog number EM30325M.

[0033] Example 1 Preparation method of 1% Apocynum Venetin Cream 1) Accurately weigh each medicine according to the following weights: 3 kg of mono- and di-stearyl glycerol 5 kg of stearyl alcohol 4 kg of liquid paraffin Ethylparaben 0.1 kg Polyoxyethylene (21) stearyl alcohol ether 2 kg Polyoxyethylene (2) stearyl alcohol ether 2 kg 2 kg of polyglycerol oleate 58 kg of ultrapure water 8 kg of glycerol 15 kg of diethylene glycol monoethyl ether Apocynum venetum 1 kg.

[0034] 2) Mix and dissolve glyceryl mono- and di-stearate, octadecanol, liquid paraffin, ethylparaben, polyoxyethylene (21) stearyl alcohol ether, polyoxyethylene (2) stearyl alcohol ether and polyglycerol oleate as component A; Weigh out ultrapure water and glycerol and mix them together as component B; Weigh out diethylene glycol monoethyl ether and apocynum venetum glycoside, mix them, and dissolve them completely to obtain component C; After heating components A and B to 85°C respectively, add components A and B while stirring to mix them thoroughly and homogenize for 3 minutes. After cooling the mixture of components A and B to 60°C, component C is added, and homogenization continues for 3 minutes. The homogenized liquid was cooled to approximately 40°C to form a paste, yielding 1% Apocynum venetum cream.

[0035] Example 2 Preparation method of 0.5% Apocynum venetum cream 1) Accurately weigh each medicine according to the following weights: 3 kg of mono- and di-stearyl glycerol 5 kg of stearyl alcohol 4 kg of liquid paraffin Ethylparaben 0.1 kg Polyoxyethylene (21) stearyl alcohol ether 2 kg Polyoxyethylene (2) stearyl alcohol ether 2 kg 2 kg of polyglycerol oleate 58 kg of ultrapure water 8 kg of glycerol 15 kg of diethylene glycol monoethyl ether Apocynum venetum 0.5 kg.

[0036] 0.5% Apocynum venetum cream was prepared according to the preparation method in Example 1.

[0037] Example 3 Effects of 0.5% and 1% Apocynum venetum cream on a hydroquinone-induced mouse model of vitiligo 1. Experimental Methods 1.1 Materials Test drug: 1% Apocynum venetum cream prepared in Example 1; Positive treatment drug: 0.05% dexamethasone acetate cream (manufacturer: Shanghai Skin Disease Hospital); Blank matrix: Cream prepared by omitting the active ingredient Apocynum venetum in Example 1; Modeling agent: 3% hydroquinone cream; Laboratory animals: SPF grade C57BL / 6J mice, 20-25mg, female, 8 weeks old, obtained from Shanghai Spaford Laboratory Animal Co., Ltd.

[0038] 2. Animal grouping and treatment Experimental animals were randomly divided into a blank control group, a model group, a positive drug group, a 0.5% Apocynum venetum treatment group, and a 1% Apocynum venetum treatment group. From day 1 to day 20 of modeling, at 9:00 AM each day, the blank control group was treated with 62.5 mg / animal of blank matrix, the model group was treated with 62.5 mg / animal of 3% hydroquinone cream, the positive control group was treated with 62.5 mg / animal of 3% hydroquinone cream, and the Apocynum venetum treatment group was treated with 62.5 mg / animal of 3% hydroquinone cream. At 5:00 PM each day, the blank control group received no treatment, the model group was treated with 62.5 mg / animal of cream matrix, the positive control group was treated with 62.5 mg / animal of dexamethasone acetate cream, the 0.5% Apocynum venetum treatment group was treated with 62.5 mg / animal of 0.5% Apocynum venetum cream, and the 1% Apocynum venetum treatment group was treated with 62.5 mg / animal of 1% Apocynum venetum cream.

[0039] On day 21 of modeling, at 9:00 AM, the blank control group was treated with the cream matrix, while the model group and the Apocynum venetum treatment group received no treatment. At 1:00 PM, blood was drawn from the orbital fossa for ELISA testing of inflammatory factors. After cervical dislocation, back tissue was harvested for HE staining and CD8 analysis. + T staining, Melan-A staining, SOX10 staining, and Masson staining were used to observe the condition of melanocytes.

[0040] 3. Experimental Results Figure 1 The study showed the skin depigmentation and repigmentation in mice after modeling and drug administration. The modeling group showed obvious depigmentation and patch formation on the back skin of the mice, indicating successful modeling. The positive drug group and the Apocynum venetum treatment group showed obvious repigmentation on the back skin, indicating that Apocynum venetum has an effective effect in treating vitiligo.

[0041] Figure 2 The expression of inflammatory factors IFN-γ, TNF-α, IL-1β, and IL-6 in mouse serum was analyzed. A significant decrease was observed in the treatment group. Based on the key role of inflammatory factor release in vitiligo, Apocynum venetum has a significant effect in reducing inflammatory factors, indicating that this natural extract that inhibits NOX activity has a prominent role in the treatment of vitiligo, an autoimmune skin disease.

[0042] Figure 3 To observe the staining status of melanocyte-specific markers (such as Melan-A and SOX10) and CD8 in the blank control group, model group, and Apocynum venetum treatment group. + T cell infiltration is clearly observed in the model group in the figure. + Increased T cell infiltration and decreased melanocyte-specific markers indicate successful modeling; correspondingly, CD8+ in the treatment group... + The significant reduction in T-cell infiltration and the increase in melanocyte-specific markers indicate that the Apocynum venetum ointment has an effective therapeutic effect on vitiligo.

[0043] Figure 4 The results showed significant differences between the model group and the treatment group, with a marked increase in melanocytes after treatment. These findings demonstrate the remarkable efficacy of the described Apocynum venetum ointment in treating vitiligo.

[0044] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. The use of Apocynum venetum or its derivatives in the preparation of drugs for the prevention and / or treatment of depigmenting skin diseases.

2. The application according to claim 1, characterized in that, The depigmentation skin diseases mentioned include vitiligo.

3. The application according to claim 1, characterized in that, The treatment of depigmented skin diseases includes at least one of the following: promoting the increase of melanocytes in the skin, promoting melanin synthesis, inhibiting melanocyte apoptosis, and protecting melanocytes from oxidative damage.

4. The application according to claim 1, characterized in that, The dosage form of the drug includes at least one of the following external dosage forms: cream, ointment, gel, lotion, liniment, and lotion.

5. The application according to claim 4, characterized in that, The drug also includes pharmaceutically acceptable excipients.

6. The application according to claim 1, characterized in that, The apocynum derivatives include apocynum salt derivatives.

7. The application according to claim 1 or 6, characterized in that, The mass percentage of apocynum or apocynum derivatives in the drug is 0.1% to 10%.

8. A cream for treating hypopigmentation skin diseases, characterized in that, The product comprises the following components in parts by weight: 0.5-1.5 parts of Apocynum venetum, 2-4 parts of glyceryl mono- and di-stearyl esters, 4-6 parts of fatty alcohol, 3-5 parts of liquid paraffin, 0.08-0.12 parts of ethylparaben, 1.8-2.2 parts of polyoxyethylene (21) stearyl alcohol ether, 1.8-2.2 parts of polyoxyethylene (2) stearyl alcohol ether, 1.8-2.2 parts of polyglycerol oleate, 55-60 parts of ultrapure water, 7-9 parts of glycerol, and 13-17 parts of diethylene glycol monoethyl ether.

9. The cream for treating depigmenting skin diseases according to claim 8, characterized in that, The components include the following parts by weight: 1 part of Apocynum venetum, 3 parts of glyceryl mono- and di-stearyl esters, 5 parts of fatty alcohol, 4 parts of liquid paraffin, 0.1 parts of ethylparaben, 2 parts of polyoxyethylene (21) stearyl alcohol ether, 2 parts of polyoxyethylene (2) stearyl alcohol ether, 2 parts of polyglycerol oleate, 58 parts of ultrapure water, 8 parts of glycerol, and 15 parts of diethylene glycol monoethyl ether.

10. The cream for treating depigmenting skin diseases according to claim 8 or 9, characterized in that, The fatty alcohols include hexadecyl alcohol and / or octadecyl alcohol.

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