Application of isovaleric acid in treatment of psoriasis

Abstract

The invention belongs to the technical field of biology, and relates to application of isovaleric acid in treatment of psoriasis. In order to solve the problems that existing psoriasis treatment drugs (such as biological agents) have side effects, the curative effect is reduced, and relapse occurs after drug withdrawal, the invention provides a psoriasis treatment scheme taking isovaleric acid as an active component, and the imiquimod-induced psoriasis-like skin lesion can be remarkably relieved by locally smearing the isovaleric acid with the concentration of 2mM; the epidermis excessive proliferation and the expression of inflammatory factors such as IL-17A, IL-17F and CXCL1 are reduced. The isovaleric acid disclosed by the invention is a small molecule compound, is simple to prepare, convenient to administrate and high in safety, fills the application blank of the isovaleric acid in the field of psoriasis treatment, and provides a new effective way for psoriasis treatment.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to the application of isovaleric acid in the treatment of psoriasis. Background Technology

[0002] Psoriasis is a common, chronic, relapsing, inflammatory skin disease, clinically characterized by well-defined red plaques covered with silvery-white scales. Its pathophysiological process is complex, involving excessive proliferation and abnormal differentiation of keratinocytes, dermal angiogenesis, and immune cell infiltration dominated by T lymphocytes (especially Th17 cells) and the release of inflammatory factors (such as IL-17, IL-22, and TNF-α).

[0003] Currently, there are various treatment strategies for psoriasis, including topical medications (such as corticosteroids, vitamin D3 derivatives, and calcineurin inhibitors), phototherapy, traditional systemic therapies (such as methotrexate, cyclosporine, and acitretin), and biologics targeting specific cytokines (such as anti-TNF-α, anti-IL-12 / 23, and anti-IL-17 drugs). Although biologics are significantly effective, their high cost, the need for injection, potential infection risks, and unknown long-term safety limit their widespread use. For mild to moderate cases, topical treatment remains the first choice; however, existing topical medications still have limitations such as insufficient efficacy, local irritation, skin atrophy, or relapse after discontinuation. Therefore, the development of novel, highly effective, safe, and easy-to-use topical medications, especially small molecule drugs based on novel mechanisms of action, has significant clinical need and market value.

[0004] Isovaleric acid, also known as 3-methylbutyric acid, is a naturally occurring short-chain fatty acid found in various plants such as valerian and angelica, as well as in human metabolism. Studies have shown that short-chain fatty acids play a role in regulating immune homeostasis and inhibiting inflammatory responses. However, to date, there have been no reports on the application of isovaleric acid in the treatment of skin diseases, particularly psoriasis, and whether it can exert a therapeutic effect by regulating key pathological processes in psoriasis remains unknown. Summary of the Invention

[0005] To address the problems existing in the prior art, this invention provides the application of isovaleric acid in the treatment of psoriasis. It is the first time that isovaleric acid has been applied to IMQ (imiquimod)-induced psoriasis-like mice. Through gross observation of the mice and statistical analysis of epidermal thickness using HE staining, it is demonstrated that isovaleric acid can alleviate the severity of the disease in psoriasis-like mice.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solutions.

[0007] This invention discloses the use of isovaleric acid in the preparation of medicaments for treating or alleviating psoriasis.

[0008] Furthermore, the dosage form of the drug is a pharmaceutically acceptable dosage form.

[0009] Furthermore, the dosage of the drug is a pharmaceutically acceptable dosage.

[0010] The present invention also discloses a pharmaceutical composition for treating or alleviating psoriasis, characterized in that it comprises a therapeutically effective amount of isovaleric acid or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable topical carrier.

[0011] Furthermore, the dosage form of the drug is a pharmaceutically acceptable dosage form.

[0012] Furthermore, the dosage of the drug is a pharmaceutically acceptable dosage.

[0013] Compared with the prior art, the beneficial effects of the present invention are as follows.

[0014] This invention is the first to discover and verify the definite efficacy of isovaleric acid in treating psoriasis, particularly in improving the superficial symptoms of psoriatic lesions and inhibiting the expression of related inflammatory factors, thus opening up a new pharmaceutical application for this known small molecule compound. Animal models have demonstrated that isovaleric acid not only improves pathological phenotypes such as epidermal hyperplasia but also specifically downregulates the expression of key psoriasis-causing factors such as IL-17A and IL-17F, suggesting that it may exert its effects by intervening in Th17-related inflammatory pathways, providing a theoretical basis for further development. Isovalleric acid is suitable for formulation into topical preparations such as creams and gels, which can be applied directly to the affected area, making it convenient to use, with high patient compliance, and avoiding the potential systemic side effects of systemic administration. Isovalleric acid is a naturally occurring short-chain fatty acid with a clear metabolic pathway in vivo, and its expected safety profile for topical application is good. The preparation process of isovaleric acid is mature, and the raw materials are readily available. Compared with complex drugs such as biological agents, it has a significant cost advantage, facilitating clinical promotion and application. Attached Figure Description

[0015] Figure 1 Verification of the therapeutic effect of isovaleric acid on an imiquimod-induced psoriasis-like mouse model. A: Gross photograph of the skin on the back of the mouse. B: Skin stained with hematoxylin and eosin (HE). C: Statistical analysis of epidermal thickness in Figure B. D: Expression of several inflammatory factors in the skin. Detailed Implementation

[0016] The present invention will now be described in further detail with reference to specific embodiments. However, this should not be construed as limiting the scope of the present invention to the following embodiments; all technologies implemented based on the content of the present invention fall within the scope of the present invention.

[0017] Unless otherwise specified, all reagents and materials used in this invention are commercially available.

[0018] I. Experimental reagents, instruments, and methods.

[0019] 1. Experimental animals: Balb / c female mice from Beijing Huafukang Company, 6-8 weeks old, weighing about 20 grams.

[0020] 2. Experimental reagents: 5% imiquimod cream (China, Sichuan Mingxinlidi Pharmaceutical Co., Ltd.), isovaleric acid (USA, MCE Company), 10% neutral formalin solution (China, Solarbio Science & Technology Co., Ltd.), ethanol (China, Solarbio Science & Technology Co., Ltd.), dewaxing and clearing agent, hematoxylin staining solution, eosin staining solution.

[0021] II. Experimental Methods.

[0022] 1. Imiquimod cream (IMQ) induces mouse model.

[0023] (1) Balb / C mice, female, 6-8 weeks old, weighing 20 grams.

[0024] (2) After feeding the mice for about a week to adapt to the new environment, the mice were divided into the IMQ group and the IMQ+isovalerate group.

[0025] (3) Shave the long hair on the back of the mouse, about 3 cm long and 2 cm wide, and then use hair removal cream to remove the downy hair. During this process, be careful not to damage the mouse's skin.

[0026] (4) Apply 62.5 mg of 5% imiquimod cream evenly to the skin on the back of each mouse for 7 days, once a day.

[0027] (5) Drug administration to mice: IVA was prepared to a concentration of 2 mM using PBS, and 200 μL was applied topically to the modeling area on the back of mice. The interval between IVA administration and IMQ modeling was 12 hours, and the process was continued for 6 days.

[0028] (6) Mouse tissue sampling: Mice were euthanized by cervical dislocation, and skin 1 cm long × 0.5 cm wide was cut from the middle of the back of the mouse. The skin was placed in 10% neutral formalin solution and fixed at room temperature for 24 h. It was then used for paraffin embedding and H&E staining.

[0029] 2. Paraffin embedding and sectioning of mouse skin tissue: (1) Fixation: Cut a piece of skin 1 cm long × 0.5 cm wide from the midline area of ​​the mouse back and place it in 10% formalin solution for 24 h at room temperature; (2) Dehydration: Mouse skin tissue was placed in 75%, 85%, 95% and 100% ethanol in sequence, and each was left to stand for 10 min; (3) Transparency: For environmentally friendly transparent agents I and II, each should be left to stand for 10 minutes; (4) Wax impregnation: Immerse in liquid paraffin and leave at room temperature for 1 hour; (5) Paraffin embedding: Place the paraffin-impregnated tissue block into a paraffin mold and cool at room temperature until the paraffin becomes solid; (6) Paraffin sections: Thin sections with a thickness of 5 μm were cut using a microtome; (7) Applying the sheet: Apply the sheet to the glass slide and place it on the heat preservation table to make it adhere firmly.

[0030] 3. Hematoxylin-eosin staining: (1) Bake in a 65℃ oven for 4-6 hours; (2) Dewaxing and hydration: Clearing agent I and clearing agent II for 15 min each, anhydrous ethanol I, anhydrous ethanol II, 95% ethanol, 85% ethanol and 75% ethanol for 5 min each, and PBS for 3 min each, followed by 3 washes. (3) Hematoxylin staining: hematoxylin staining solution for 5 min, then rinse slowly with running water for 5 min; (4) Hydrochloric acid-alcohol differentiation: immerse in 1% hydrochloric acid-alcohol for 5 seconds, then rinse slowly with running water for 1 minute; (5) Eosin staining: Place in eosin staining solution for 1 min, then wash with distilled water for 10 s; (6) Dehydration and clearing: 5% ethanol for 1 min, 85% ethanol for 1 min, 95% ethanol for 1 min, anhydrous ethanol II for 5 min, anhydrous ethanol I for 5 min, clearing agent II for 15 min, clearing agent I for 15 min; (7) Neutral resin sealing.

[0031] 4. Statistical Analysis: GraphPad Prism 8.0 software was used for analysis and graphing. T-tests were used for statistical analysis between the two groups. A p-value less than 0.05 was considered statistically significant. The results of epidermal thickness were quantified using Image ProPlus 6.0 software. In the statistical tests of this experiment, * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001, and ns indicates no statistical difference. All data are expressed as mean ± standard deviation.

[0032] 5. Qpcr.

[0033] RNA extraction and quantitative real-time PCR: mRNA was collected using the miRNeasy Mini Kit (Qiagen, Germany) and measured using an ND1000 nanometer. cDNA was synthesized using the GoScript reverse transcription kit (Promega, USA). RT-PCR was then performed using the RTSYBR Green qPCR Mastermix (Promega, USA), and detected using a 7900HT Fast Real-Time PCR System (Applied Biosystems, USA). The reaction conditions were set at 95°C for 2 min, followed by a 15 s extension at 95°C and a 1 min extension at 60°C for 40 cycles. Melting curves for each PCR product were calculated to confirm synthesis specificity.

[0034] III. Experimental Results.

[0035] As shown in the figure, gross observation of the skin on the backs of mice revealed that, compared to the control group, the isovaleric acid (IVA) group showed reduced skin lesions. HE staining also showed a decrease in epidermal thickness and inflammatory factors in the isovaleric acid group.

[0036] The above description is merely a preferred embodiment of the present invention and is not intended to limit the patent scope of the present invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. The use of isovaleric acid in the preparation of drugs for the treatment or relief of psoriasis.

2. The application according to claim 1, characterized in that, The dosage form of the drug is pharmaceutically acceptable.

3. The application according to claim 1, characterized in that, The dosage of the drug is pharmaceutically acceptable.

4. A pharmaceutical composition for treating or alleviating psoriasis, characterized in that, It contains a therapeutically effective amount of isovaleric acid or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable topical carrier.

5. The pharmaceutical composition according to claim 4, characterized in that, The dosage form of the drug is pharmaceutically acceptable.

6. The pharmaceutical composition according to claim 4, characterized in that, The dosage of the drug is pharmaceutically acceptable.

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