Application of Daurisoline in preparation of psoriasis treatment medicine and medicine

Abstract

The invention discloses application of Daurisoline in preparation of a medicine for treating psoriasis and the medicine, relates to the technical field of preparation of the medicine for treating the psoriasis, provides new application of the Daurisoline and a new medicine for treating the psoriasis, and provides a new strategy for providing a new treatment method and a new treatment medicine for clinical intervention of the psoriasis.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to application of Daurisoline in preparing a drug for treating psoriasis and the drug. Background Art

[0002] Psoriasis is a common immune-mediated chronic inflammatory skin disease, accompanied by hyperkeratosis and multiple inflammatory reactions. The global incidence is about 3%, with about 25 million patients in Europe and the United States. The incidence of psoriasis in my country is 0.72%, with about 10 million patients, and 100,000 new patients each year. The disease is obviously seasonal at the onset, mostly severe in winter and mild in summer. The whole body can be affected, but it usually occurs on the scalp, finger (toe) nails, the extensor side of the limbs (especially the anterior tibia of the lower leg), and the lumbar sacral region. It is generally symmetrically distributed, and the clinical manifestations are red plaques covered with thick silvery white scales. The main skin pathological characteristics of psoriasis are: epidermal hyperplasia, obvious proliferation of keratinocytes, increased and expanded angiogenesis, and obvious inflammatory infiltration. According to its clinical manifestations and pathological characteristics, it is mainly divided into common psoriasis, pustular psoriasis, arthritis psoriasis, and erythrodermic psoriasis, among which common psoriasis is the most common, accounting for more than 97% of all patients. Psoriasis is a stubborn disease that is prone to recurrence, which causes a heavy burden on patients' lives and mental distress. However, the specific pathogenesis of psoriasis is still unclear. Studies have shown that genetic factors, environmental factors, and autoimmune factors jointly lead to the occurrence and development of psoriasis.

[0003] Treatment of psoriasis is generally divided into systemic treatment, local treatment and physical therapy. Drugs for systemic treatment of psoriasis include: retinoic acid, methotrexate, ethyl corydalis, etc. Studies have shown that systemic treatment of psoriasis is effective and easy to use, but it will cause certain adverse reactions such as nephrotoxicity, hypertension, hepatotoxicity, and hyperlipidemia. Local therapies include: topical hormones such as halometasone, tar, anthralin, retinoic acid cream, tamoxifen, vitamin D3 derivatives, etc. Physical therapy: Ultraviolet irradiation therapy is currently the best treatment for psoriasis patients, but its treatment method is relatively unchanged, and it has certain negative effects on normal skin during treatment. In addition, due to the long treatment process of psoriasis, some patients will develop drug resistance after long-term use of a certain drug. Therefore, it is urgent to find a new therapeutic drug that can bring new ideas for the treatment of psoriasis. Summary of the invention

[0004] In view of this, the present invention provides a new use of Daurisoline and a new drug for treating psoriasis, providing a new treatment method and a new strategy for clinical intervention of psoriasis.

[0005] The present invention solves the above technical problems by the following technical means:

[0006] The invention discloses application of Daurisoline in preparing a drug for treating psoriasis.

[0007] According to the above application, the concentration of Daurisoline in the medicine prepared by the present invention is 40 μM.

[0008] Furthermore, the dosage form of the drug is a solution, lotion, liniment, ointment, plaster, paste or patch.

[0009] Furthermore, the medicine is an ointment, which consists of Daurisoline and a coating base.

[0010] Since the medicament of the present invention is used to treat psoriasis, it is preferably in the form of an ointment, which is convenient for application and wiping.

[0011] Furthermore, the coating base includes vaseline, and the concentration of Daurisoline in the drug is 40 μM.

[0012] Furthermore, the medicine also includes one or more of a preservative, a buffer, a disintegrant, an antioxidant, a suspending agent, a colorant and an excipient.

[0013] Of course, excipients can be added to extend the drug's storage stability, shelf life, ease of use, etc.

[0014] Beneficial effects of the present invention:

[0015] Daurisoline treatment can significantly alleviate the IMQ-induced psoriasis-like dermatitis phenotype in mice, reduce inflammatory cell infiltration, significantly reduce scaling and redness of the mouse skin, and alleviate the level of epidermal proliferation. It has an inhibitory effect on the secretion of related inflammatory factors and is effective in the treatment of psoriasis, providing a new strategy for the clinical treatment of psoriasis. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 :Pictures of experimental skin lesions on the back of psoriasis mice and cell staining;

[0017] Figure 2 :Daurisoline inhibits the secretion of inflammatory cytokines in Mix-induced psoriasis-like HaCaT cells;

[0018] Figure 3 :Daurisoline inhibits the secretion of inflammatory cytokines in Mix-induced psoriasis-like HaCaT cells;

[0019] Figure 4 :The results of applying Daurisoline to alleviate the inflammatory phenotype associated with IMQ-induced psoriasis mice. DETAILED DESCRIPTION

[0020] The present invention will be described in detail below with reference to the embodiments and accompanying drawings:

[0021] Daurisoline, Chinese name: Daurisoline, is a chemical substance with the chemical formula C 37 H 42 N2O6, molecular weight 610.74, structural formula Figure 1 shown.

[0022] Example 1: Mouse Experiment

[0023] 1. Construction of animal experimental model: The C57 male mice used in the present invention were purchased from the Experimental Animal Center of Air Force Medical University and were raised in the SPF barrier environment of the center.

[0024] The mice were randomly divided into four groups: WT control group (coating matrix), WT treatment group (coating matrix + Daurisoline), IMQ control group (IMQ + coating matrix), and IMQ experimental group (IMQ + coating matrix + Daurisoline).

[0025] The coating base was composed of the same dose of carrier - vaseline (Vaseline; Unilever, Englewood Cliffs, USA).

[0026] The concentration of Daurisoline in the WT treatment group and the IMQ experimental group was 40 μM.

[0027] The specific treatment of wild mice is as follows:

[0028] The backs of C57 mice were depilated after two days, and different drugs were treated on the backs of the mice every morning for five consecutive days (the WT control group was smeared with the smear matrix, and the WT treatment group was smeared with the smear matrix mixed with Daurisoline), and the smear was applied evenly;

[0029] The specific treatment of the imiquimod-induced psoriasis mouse model is as follows:

[0030] The back of C57 mice was depilated. Two days later, 20 mg of IMQ (5% IMQ, INova Pharmaceuticals, Chatswood, NSW, Australia) was applied to the back of the mice once every morning for five consecutive days to prepare an imiquimod-induced psoriasis mouse model. At the same time, different drug treatments were applied in the afternoon (the IMQ control group was applied with the application matrix, and the IMQ experimental group was applied with the application matrix mixed with Daurisoline). The application was evenly applied.

[0031] The above treatments were applied continuously for 5 days before the mice were killed. During this period, the incidence of psoriasis on the back of the mice was observed and photographed, and the back skin of the mice was taken for H&E staining.

[0032] 2. Result analysis:

[0033] The results are as follows Figure 2 The results are analyzed as follows:

[0034] from Figure 2 It can be seen that for mice without IMQ modeling, the application of the matrix and Daurisoline had no effect on the back skin of the mice. For psoriasis mice with successful IMQ modeling, the topical application of the matrix mixed with Daurisoline can significantly alleviate the erythema and swelling of the skin of psoriasis mice, reduce scales, and reduce the thickness of the skin epidermis. The comparison of staining results shows that the inflammatory cell infiltration of mice in the IMQ experimental group was reduced.

[0035] Example 2: qRT-PCR detection of cells and mouse tissues

[0036] 1. Cell sample preparation:

[0037] The cells used in this example are human keratinocytes HaCaT. HaCaT cells were seeded in 6-well plates, with 1×10 6 Each group of cells had 3 replicate wells and was placed in a 37°C incubator containing 5% CO2 for routine culture. Mixed cytokines were used to simulate the cell state of psoriasis.

[0038] The experiment was divided into Control group (Daurisoline) and Mix group (Daurisoline + 50ng / mL IL-17A, 20ng / mL IL-22, 50ng / mL TNF-α, 20ng / mL IL-1α and 20ng / mL oncostatin M (OSM) mixed cytokine Mix) for corresponding stimulation. The Daurisoline added to the Control group and Mix group was designed with different concentration gradients, namely 0μM, 3.125μM, 6.25μM, 12.5μM, and 25μM, respectively. Samples were collected after 24 hours for qRT-PCR detection.

[0039] 2. Mouse tissue sample preparation:

[0040] The skin lesions of the mice in each group in Example 1 were cut into pieces and ground into powder to collect samples for qRT-PCR detection.

[0041] 3. qRT-PCR detection:

[0042] RNA was extracted from tissues and cells using the guanidine isothiocyanate-phenol-chloroform extraction method, and RNA was quantified using the reverse transcription kit PrimeScript TM RT reagent Kit is used to convert the DNA into cDNA.

[0043] According to the quantitative PCR kit TB Premix Ex Taq TM Instructions: Prepare the PCR reaction solution in a PCR tube and perform qRT-PCR experiment on the CFX384 Real-Time PCR Detection System instrument, and then perform relevant inflammatory phenotype detection.

[0044] 4. Result analysis:

[0045] (1) Figure 3 As shown, Daurisoline can significantly inhibit the mRNA expression levels of inflammatory cytokines, chemokines and antimicrobial peptide-related factors in the psoriasis-like HaCaT cell model induced by mixed cytokine Mix in a dose-dependent manner.

[0046] (2) Figure 4 As shown, Daurisoline mixed with the application matrix can significantly reduce the mRNA expression levels of inflammatory cytokines, chemokines, antimicrobial peptides and proliferation-related factors in psoriasis mice, and increase the mRNA expression levels of differentiation and barrier-related factors in psoriasis mice.

[0047] In summary, the present invention uses psoriasis-like cell models and animal models to find that Daurisoline can significantly inhibit the expression and secretion of related inflammatory factors and thus inhibit psoriasis-like skin lesions, providing a causal treatment method for clinical intervention and a new strategy for the treatment of psoriasis.

[0048] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the present invention. Although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the purpose and scope of the technical solutions of the present invention, which should be included in the scope of the claims of the present invention. The techniques, shapes, and structural parts not described in detail in the present invention are all known technologies.

Claims

1. Application of Daurisoline in the preparation of drugs for the treatment of psoriasis.

2. The use according to claim 1, characterized in that: The concentration of Daurisoline in the drug was 40 μM.

3. The medicine prepared by the application according to any one of claims 1 or 2, characterized in that: The dosage form of the drug is solution, lotion, liniment, ointment, plaster, paste or patch.

4. The medicine prepared by the application according to claim 3, characterized in that: The medicine is an ointment, which consists of Daurisoline and a coating base.

5. The medicine prepared by the application according to claim 4, characterized in that: The coating base includes petrolatum.

6. The medicine prepared by the application according to claim 3, characterized in that: The medicament may further comprise one or more of a preservative, a buffer, a disintegrant, an antioxidant, a suspending agent, a colorant and an excipient.

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