TSLP expression level regulator, medicine for treating psoriasis and application

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a TSLP expression level regulator, a medicine for treating psoriasis and application. The invention provides a TSLP expression level regulator, the TSLP expression level regulator is a TSLP inhibitor, the inhibitor is composed of a chlorguanidine substance and bacillus subtilis, and the mass ratio of the chlorguanidine substance to the bacillus subtilis is (20-50) mg: (1-1.5) g. The invention finds that the bacillus subtilis can enhance the effect of treating psoriasis by the chlorguanidine, the chlorguanidine and the bacillus subtilis have a synergistic effect, and the composition of the chlorguanidine and the bacillus subtilis can well inhibit the TSLP expression level.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and specifically relates to a TSLP expression level regulator and a drug for treating psoriasis and its application. Background Art

[0002] Psoriasis is a chronic inflammatory skin disease that is difficult to treat and recurs repeatedly. In recent years, targeted drugs for psoriasis have developed rapidly, as follows.

[0003] TNF-α inhibitors: Etanercept, infliximab, adalimumab and pecilizumab have been used clinically for many years with definite efficacy, but some patients may experience problems such as immunogenic reactions and increased risk of infection.

[0004] IL-17 inhibitors: Secukinumab, Ixekizumab, etc. IL-17 inhibitors have shown good efficacy and safety in the treatment of psoriasis, especially for patients with moderate to severe plaque psoriasis. The current research focus is on exploring its optimal application in different types of psoriasis, as well as combination treatment strategies with other drugs.

[0005] IL-23 inhibitors: ustekinumab, guselkumab, etc. IL-23 inhibitors can effectively control psoriasis inflammation and have good long-term efficacy and safety. Future research directions include further optimizing the drug dosing regimen and developing more new inhibitors targeting different IL-23 subunits.

[0006] IL-36 inhibitors: Pesorimab is approved for the treatment of pustular psoriasis, providing a new option for the treatment of this particular type of psoriasis. Researchers are studying its potential application value in other types of psoriasis and the possibility of combining it with other drugs.

[0007] All of the above inhibitors have targeted effects and provide a reliable approach for the treatment of psoriasis. However, due to the large number of psoriasis types and patients, it is necessary to continue to develop new therapeutic targets or therapeutic drugs.

[0008] Thymic stromal lymphopoietin, abbreviated as TSLP, is an IL-7-like inflammatory factor. Studies have shown that TSLP is associated with a variety of diseases. For example, in allergic diseases such as asthma, atopic dermatitis, and food allergies, TSLP expression is usually upregulated; in autoimmune diseases such as rheumatoid arthritis and systemic lupus erythematosus, TSLP expression may also be abnormal; in a variety of tumors, TSLP expression is closely related to the occurrence, development, and prognosis of tumors. In cells, the expression of TSLP is regulated by a variety of factors. When stimulated by exogenous antigens such as bacteria, viruses, and Toll-like receptor agonists, TSLP in the blood will increase rapidly. Other studies have shown that dendritic cells, abbreviated as DC, are the main cell source of TNF-α and IL-23, and DC activation plays an important role in the initial stage of psoriasis. DC is the main effector cell of TSLP, and the expression level of TSLP is also related to psoriasis.

[0009] Therefore, from the perspective of treating diseases, it is necessary to develop a TSLP expression level regulator. Summary of the invention

[0010] In order to solve the above technical problems, the present invention provides a TSLP expression level regulator.

[0011] The purpose of the present invention is to provide a TSLP expression level regulator, which is a TSLP inhibitor. The inhibitor consists of chlorguanil and Bacillus subtilis, and the mass ratio of the two is 20mg~50mg:1g~1.5g.

[0012] The structural formula of proguanil is Figure 1 , is a TSLP inhibitor, which inhibits the secretion of TSLP from the patient's mast cells and has an improving effect on atopic dermatitis. When treating atopic dermatitis, the dosage is about 50mg~500mg. However, atopic dermatitis is mainly caused by allergic reactions. Atopic dermatitis is different from psoriasis. The pathogenesis of psoriasis is complex, so the treatment of psoriasis is relatively difficult. The present invention mainly discusses the therapeutic effect of proguanil on psoriasis. Bacillus subtilis is a probiotic that can regulate intestinal microecology. The existing technology is often used to improve digestion ability. The present invention combines proguanil and Bacillus subtilis to synergistically enhance the effect to inhibit the expression of TSLP, thereby inhibiting psoriasis caused by TSLP activation of DC.

[0013] Preferably, the TSLP expression level regulator, the proguanil-like substance is proguanil or a pharmaceutically acceptable salt thereof, such as the proguanil hydrochloride and proguanil sulfate, wherein the formation of hydrochloride and sulfate increases the water solubility of the drug, which is very important for the preparation of drug preparations.

[0014] Preferably, the TSLP expression level regulator and the inhibitor are used to prepare a drug for treating psoriasis.

[0015] Preferably, the TSLP expression level regulator is used for psoriasis-like skin inflammation caused by abnormal TSLP expression. More specifically, the psoriasis is psoriasis-like skin inflammation caused by upregulated TSLP expression induced by IMQ.

[0016] Preferably, the TSLP expression level regulator, the inhibitor is used to inhibit the JAK1 / SYK / TRAF6 / NF-κB pathway.

[0017] Preferably, the TSLP expression level regulator and the inhibitor are used to reduce the expression level of IL-17A.

[0018] Preferably, the TSLP expression level regulator and the inhibitor are used to inhibit DC activity.

[0019] Preferably, the TSLP expression level regulator, the inhibitor is used to inhibit CD4 + T cells differentiate into Th17 cells.

[0020] The present invention provides a drug for treating psoriasis, which uses the proguanil substance as an active ingredient and is prepared with a first excipient acceptable in pharmaceutical science to prepare a proguanil substance drug; The Bacillus subtilis is used as an active ingredient and a pharmaceutically acceptable second auxiliary material is added to prepare a Bacillus subtilis medicine; The mass ratio of chlorguanil substances to Bacillus subtilis is 20mg~50mg:1g~1.5g, calculated based on the mass of chlorguanil substances in chlorguanil drugs and the mass ratio of Bacillus subtilis in Bacillus subtilis drugs.

[0021] Preferably, the drug for treating psoriasis is an oral preparation.

[0022] Preferably, the first excipient and the second excipient used in the oral preparation are the same, and are at least one of a filler, a disintegrant, a lubricant and a solubilizing agent.

[0023] The filler includes at least one of microcrystalline cellulose, lactose, and starch, wherein the mass percentage of microcrystalline cellulose in the oral preparation is 20% to 90%, the mass percentage of lactose in the oral preparation is 10% to 70%, and the mass percentage of starch in the oral preparation is 5% to 30%.

[0024] The disintegrant comprises at least one of sodium carboxymethyl starch and cross-linked polyvinylpyrrolidone, wherein the mass percentage of sodium carboxymethyl starch in the oral preparation is 2% to 8%, and the mass percentage of cross-linked polyvinylpyrrolidone in the oral preparation is 1% to 5%.

[0025] The lubricant includes at least one of magnesium stearate and talc, wherein the mass percentage of magnesium stearate in the oral preparation is 0.1% to 1%, and the mass percentage of talc in the oral preparation is 0.5% to 2%.

[0026] The cosolvent includes at least one of polyethylene glycol and sodium lauryl sulfate, wherein the mass percentage of polyethylene glycol in the oral preparation is 5% to 30%, and the mass percentage of sodium lauryl sulfate in the oral preparation is 0.1% to 2%.

[0027] It should be noted that the above excipients are only specific embodiments introduced for illustrative purposes. Oral preparations made by those skilled in the art using other excipients and the active ingredients of the present invention, or drugs in other dosage forms, all fall within the inventive concept of the present application.

[0028] Compared with the prior art, the present invention has the following beneficial effects: The present invention first discovered that the inhibitor composed of proguanil and Bacillus subtilis can reduce the expression level of TSLP. Bacillus subtilis can enhance the effect of proguanil in treating psoriasis, and the two have a synergistic effect. In order to avoid proguanil inhibiting the activity of Bacillus subtilis, the mass ratio of proguanil to Bacillus subtilis is 20mg~50mg:1g~1.5g.

[0029] The recommended time interval for using proguanil to inhibit Bacillus subtilis is 2h~4h.

[0030] The research results of the present invention show that the inhibitor composed of proguanil and Bacillus subtilis can inhibit the JAK1 / SYK / TRAF6 / NF-κB pathway, inhibit DC activity, reduce the expression level of IL-17A, inhibit CD4 + T cells differentiate into Th17 cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is the structural formula of proguanil.

[0032] Figure 2 The mean fluorescence intensity detection results of MHCII-FITC, CD80-FITC and CD86-FITC; Among them, A is MHCII, B is CD80-FITC, and C is CD86-FITC.

[0033] Figure 3 The relative expression detection results of JAK1, SYK, NF-κB p65, and TRAF6; Among them, A is p-JAK1 / JAK1, B is p-SYK / SYK, C is p-NF-κB p65 / NF-κB p65, and D is TRAF6 / GAPDH.

[0034] Figure 4 Different treatments for CD4 + The impact of T cell differentiation into Th17 cells; Among them, A is the relative expression of IL17A / β-actin, and B is the expression level of IL-17A.

[0035] Figure 5 The expression levels of p-STAT3 and RORγt-related proteins under different treatments; Among them, A is the relative expression level of p-STAT3 / STAT3, and B is the relative expression level of RORγt / β-actin.

[0036] Figure 2~Figure 5 Different lowercase letters above the bars represent significant differences between the groups. P <0.01. DETAILED DESCRIPTION

[0037] In order to enable those skilled in the art to better understand and implement the technical solution of the present invention, the present invention is further described below in conjunction with specific embodiments and drawings.

[0038] In the description of the present invention, unless otherwise specified, all reagents used are commercially available and all methods used are conventional techniques in the art.

[0039] Example 1 A TSLP expression level regulator is a TSLP inhibitor, the inhibitor consists of proguanil and Bacillus subtilis, the mass ratio of proguanil to Bacillus subtilis is 20mg:1g, and the use time interval between proguanil and Bacillus subtilis is 2h.

[0040] Experiment 1: Effect of TSLP expression level regulator on activation of mouse bone marrow-derived dendritic cells, where the English abbreviation of mouse bone marrow-derived dendritic cells is BMDC.

[0041] 1. Reagents PBS buffer: KCl 0.2 g, NaCl 8 g, KH2PO4 0.2 g, Na2HPO4·12H2O 2.89 g, and deionized water to make up to 1000 mL.

[0042] RPMI-1640 culture medium: from Gibco, USA.

[0043] Fetal bovine serum: from Thermo Fisher Scientific, USA.

[0044] Fetal bovine serum albumin: English abbreviation BSA, comes from Beijing Solarbio Company.

[0045] BCA protein quantification kit: Shanghai Biotech Biotechnology Co., Ltd.

[0046] Mouse IL-4 recombinant protein: English abbreviation IL-4, from Suzhou Jinan Bioprotein Technology Co., Ltd.

[0047] Anti-CD11c-PE antibody: full name hamster anti-mouse CD11c-coupled PE monoclonal antibody, from Santa Cruz Biotechnology (Shanghai) Co., Ltd.

[0048] Anti-MHCII-FITC antibody: full name anti-mouse MHCII-conjugated FITC monoclonal antibody, from Abcam, USA.

[0049] Rabbit anti-mouse GAPDH polyclonal antibody: from Hangzhou Huaan Biotechnology Co., Ltd.

[0050] Anti-CD80-FITC antibody: full name anti-mouse CD80-conjugated FITC monoclonal antibody, from Abcam, USA.

[0051] Anti-CD86-FITC antibody: full name anti-mouse CD86-conjugated FITC monoclonal antibody, from Abcam, USA.

[0052] RIPA protein lysis buffer: abbreviated as RIPA lysis buffer, comes from Beijing Solarbio Company.

[0053] Rabbit anti-mouse JAKl Phospho-Tyr1022 polyclonal antibody: from Signalway Antibody Company, USA.

[0054] Rabbit anti-mouse SYK Phospho-Tyr348 polyclonal antibody: from Signalway Antibody Company, USA.

[0055] Rabbit anti-mouse NFkB p65 Phospho-Ser536 polyclonal antibody: from Cell Signaling Technology, USA.

[0056] Rabbit anti-mouse TRAF6 polyclonal antibody: from Wuhan ABclonal Biotechnology Co., Ltd.

[0057] TSLP protein: It is the mouse TSLP recombinant protein, which comes from Suzhou Jinan Bioprotein Technology Co., Ltd.

[0058] GM-CSF: It is the mouse GM-CSF recombinant protein, which comes from Suzhou Jinan Bioprotein Technology Co., Ltd.

[0059] Proguanil: from Aladdin, CAS number 500-92-5.

[0060] Bacillus subtilis: 10 billion CFU / g, sourced from Weikehaisi (Shandong) Bioengineering Co., Ltd.

[0061] Imiquimod: full name Imiquimod cream.

[0062] TSLP expression level regulator: the formulation of Example 1.

[0063] 2. Methods and Results (1) Flow cytometry detection of the effects of different treatments on BMDC activation Femurs and tibiae of 6-week-old mice were washed with PBS buffer. After erythrocyte lysis, mixed cells from the limbs were washed in RPMI-1640 medium and then seeded in 6-well culture dishes at 1 × 10 cells per well. 7 DCs were induced for 8 consecutive days by adding RPMI-1640 medium containing 10% fetal bovine serum, 20ng / mL IL-4 and 20ng / mL granulocyte-macrophage colony-stimulating factor, and the supplementary medium was replaced every day. Granulocyte-macrophage colony-stimulating factor is called granulocyte-macrophage colony-stimulating factor, or GM-CSF for short.

[0064] On the third day of induction, when the cells adhered firmly to the wall, the culture medium was removed and replaced with new culture medium to remove the non-adherent monocytes and red blood cells; on the sixth day of induction, a large number of cells were observed to be suspended, or between the adherent and suspended states. At this time, the suspended cells were collected and the adherent cells were gently blown off with a gun tip. The target DC cells were blown off. The collected suspended DC cells were placed in a new culture medium; 20ng / mL IL-4 and 20ng / mL GM-CSF were continued to be given for 48h of induction to finally obtain BMDC.

[0065] After induction, BMDCs cultured in vitro were treated with 0 ng / mL TSLP protein as a control group, 10 ng / mL TSLP protein as a TSLP treatment group, and 10 ng / mL TSLP protein + the expression level regulator of Example 1 for 48 h. The expression level regulator of Example 1 was used to add 20 ng / mL proguanil while adding TSLP protein, and 1 μg / mL Bacillus subtilis was added 2 h later, which was recorded as the drug treatment group.

[0066] Anti-CD11c-PE, anti-MHC II-FITC, anti-CD80-FITC and anti-CD86-FITC antibodies were used to identify the maturation and activation levels of BMDCs. Flow cytometry was used to detect the fluorescence intensity of BMDC maturation-related protein markers MHCII, CD80 and CD86. Three parallels were performed in each group, and the test results were subjected to a one-way ANOVA test. The results are shown in Figure 2 Compared with the control group, the fluorescence intensity of MHCII, CD80 and CD86 in the TSLP-treated group increased significantly. P <0.01, while the fluorescence intensity of the drug-treated group was significantly lower than that of the TSLP-treated group. P <0.01, indicating that the inhibitor of Example 1 can inhibit the effect of TSLP.

[0067] (2) Western blot detection of the regulatory effects of different treatments on pathway proteins After the induction was completed as described in (1), 0 ng / mL TSLP protein was given as a control group, 10 ng / mL TSLP protein was given as a TSLP treatment group, and 10 ng / mL TSLP protein + the expression level regulator of Example 1 were given to the BMDC cultured in vitro for 48 hours. Among them, the expression level regulator of Example 1 was used to add 20 ng / mL proguanil while adding TSLP protein, and 1 μg / mL Bacillus subtilis was added 2 hours later, which was recorded as the drug treatment group. Three parallels were made for each group, and the test results were subjected to a one-way ANOVA test.

[0068] Collect cell samples with different treatments, add RIPA lysis buffer, add 5μL protease inhibitor mixture, 5μL PMSF and 5μL phosphatase inhibitor mixture to 1mL RIPA lysis buffer, and place in ice for lysis for 30min; centrifuge at 12000rpm, 4℃ for 15min, and aspirate the supernatant into a new 1.5mL EP tube; take part of the protein for BCA quantitative protein concentration: gradient dilution of fetal bovine serum albumin standard to 15μg / μL, 10μg / μL, 5μg / μL, 2.5μg / μL, 1μg / μL, prepare BCA working solution, BCA working solution includes A solution: BCA alkaline solution, B solution: 4% volume fraction of copper sulfate, mix A solution and B solution at a volume ratio of 50:1, add 5μL protein to a 96-well plate, add 200μL BCA mixture, place at 37℃ for 30min, cool to room temperature, measure the absorbance at 562nm with an enzyme reader, and make a standard curve. According to the absorbance of the sample to be tested, compare the standard curve and calculate the protein concentration. The volume of the solution containing 50 μg of protein is the loading amount. The loading sample is placed in a 0.5 mL centrifuge tube and 5× SDS loading buffer is added to a final concentration of 1×. The sample is heated at 100°C for 10 minutes for denaturation.

[0069] When JAK1 and SYK are activated, the levels of phosphorylated JAK1 Y1022 and phosphorylated SYK Y348 will increase. The present invention determines whether JAK1 and SYK are activated by quantitatively analyzing the relative expression levels of p-JAK1, p-SYK, p-NF-κB p65 and TRAF6, wherein NF-κB p65 is an important member of the NF-κB family.

[0070] Results Figure 3 Compared with the control group, p-JAK1, p-SYK, p-NF-κB p65 and TRAF6 in the TSLP-treated group increased significantly. P <0.01, while p-JAK1, p-SYK, p-NF-κB p65 and TRAF6 in the drug-treated group were significantly decreased compared with those in the TSLP-treated group. P <0.01, indicating that the inhibitor of Example 1 can inhibit the effect of TSLP on the JAK1 / SYK / TRAF6 / NF-κB pathway.

[0071] (3) Effects of different treatments on CD4 + Effect of T cells on differentiation into Th17 cells The present invention investigates the effect of TSLP on CD4 + The effect of T cells on their differentiation into Th17 cells.

[0072] The experimental method is to separate mouse spleen cells and obtain mouse CD4+ T lymphocyte isolation kit to obtain CD4 + T lymphocytes, CD4 + T cells were cultured with BMDC cell culture supernatant, BMDC culture medium stimulated by TSLP, and BMDC culture medium co-incubated with TSLP and JAK-SYK pathway inhibitor; the JAK-SYK pathway inhibitor was the TSLP expression level regulator of Example 1. qPCR and ELISA were used to detect the expression level of IL-17A.

[0073] The concentration of BMDC+TSLP stimulation was as follows: CD4 + T cells.

[0074] The concentration of the drug for BMDC+Example 1 stimulation was: 100 ng / mL TSLP protein + TSLP expression level regulator of Example 1 were used for co-treatment for 72 hours, and then BMDC cell culture medium was used for CD4 + T cells. The method of using the TSLP expression level regulator of Example 1 is to add 200 ng / mL proguanil while adding TSLP protein, and then add 10 μg / mL Bacillus subtilis 2 hours later.

[0075] BMDC group: CD4 + T cells were cultured using BMDC cell culture medium.

[0076] CD4 without any stimulation + The T cell group served as the control group.

[0077] Each group was tested in triplicate and the results were subjected to one-way ANOVA test.

[0078] Results Figure 4 . Figure 4 The results showed that the treatment of BMDC+Example 1 could inhibit and reduce the expression levels of IL-17A and IL-17, which was significantly different from the other groups. P <0.01.

[0079] (4) Detection of p-STAT3, the main transcription factor for Th17 cell differentiation, and RORγt protein expression levels Western blot detection of CD4 + The phosphorylation level of STAT3 protein and the expression level of RORγt protein, which are the hallmark proteins of Th17 cell differentiation in T cells, are both important nuclear transcription factors of Th17 cells.

[0080] The concentration of BMDC+TSLP stimulation was as follows: CD4 + T cells.

[0081] BMDC + TSLP + Example 1 stimulated drug concentrations: CD4 + T cells were cultured in BMDC cell culture medium treated with 100 ng / mL TSLP protein and the TSLP expression level regulator of Example 1 for 72 hours. P <0.01. The usage of the TSLP expression level regulator in Example 1 is to add 200 ng / mL proguanil while adding TSLP protein, and then add 10 μg / mL Bacillus subtilis 2 hours later.

[0082] BMDC group, CD4 + T cells were cultured using BMDC cell culture medium.

[0083] CD4 without any stimulation + The T cell group served as the control group.

[0084] Each group was tested in triplicate and the results were subjected to one-way ANOVA test.

[0085] Results Figure 5 . Figure 5 The results showed that the treatment of BMDC+Example 1 could reduce the expression levels of p-STAT3 and RORγt proteins, which was significantly different from the results of other groups. P <0.01.

[0086] Experiment 2: Experiment on the treatment of psoriasis Eight-week-old BalB / C male mice were selected and adaptively raised for a period of time to ensure that the mice were in good condition and had no obvious diseases or abnormal behaviors.

[0087] The mice were randomly divided into 3 groups, 4 mice in each group, and the specific treatment process was as follows: Imiquimod group: This group was used to construct a skin inflammation model. Before modeling, the back of the mouse was carefully shaved with a shaver in an area of ​​about 2×3 cm. 2 The hair of the mice was shaved, and the skin of the mice was damaged as much as possible during the operation. After shaving, imiquimod cream was evenly applied to the back skin of the mice at a dose of 62.5 mg per day for 9 consecutive days. During this period, the back skin of the mice was observed, and the model was considered successful when obvious erythema and scaling appeared.

[0088] Example 1 group: The back of the mice was shaved before modeling, and the area was the same as that of the imiquimod group. Six days before imiquimod induction, the mice were gavaged once a day with the TSLP expression level regulator of Example 1, first gavaged with 100 μL of 200 ng / mL proguanil, and then gavaged with 100 μL of 10 μg / mL Bacillus subtilis 2h later. Starting from the seventh day, imiquimod cream was applied to the back skin of the mice every day in the same dosage and frequency as the imiquimod group, that is, 62.5 mg per day for each mouse, and applied continuously for 9 days.

[0089] Control group: As a normal control, no special treatment related to modeling was performed, and the same dose of vaseline was applied to the back.

[0090] Each group was tested in triplicate and the results were subjected to one-way ANOVA test.

[0091] The BALB / c mouse psoriasis animal model was established with imiquimod cream. After the modeling, the severity of psoriasis lesions on the mouse skin, including erythema, scaling, and lesion thickness, were scored and compared. The results showed that on the 9th day of modeling, imiquimod could induce psoriasis-like lesions in BALB / c mice, and after pretreatment with the TSLP expression level regulator of Example 1, the expression of TSLP was reduced, which could reduce the severity of the disease.

[0092] The scoring criteria are as follows: Erythema: no obvious erythema on the epidermis was scored as 0 points; a small amount of erythema with light red color was scored as 1 point; a large amount of erythema with red color was scored as 2 points; a large amount of erythema with dark red color was scored as 3 points; a large amount of erythema with extremely dark color was scored as 4 points.

[0093] Scaling: No obvious scaling was scored as 0 points; small thin scales were visible on part of the skin surface, scored as 1 point; thick flaky scales were visible on most of the skin surface, scored as 2 points; most of the skin was covered with thick large scales, scored as 3 points; the entire skin was completely covered with thick scales, scored as 4 points.

[0094] Thickness: 0 points if the thickness is basically the same as normal skin, 1 point if the thickness of the skin surface is slightly higher than the normal skin surface, 2 points if the thickness is moderately raised, 3 points if the thickness is obviously raised, and 4 points if the thickness is highly raised.

[0095] The scores of each group are shown in Tables 1 to 3. The results of Tables 1 to 3 show that after treatment with the TSLP expression level regulator of Example 1, the scores were lower than those of the imiquimod group, indicating that the TSLP expression level regulator of Example 1 can treat psoriasis.

[0096] Table 1 Erythema scores of each group Table 2 Squamous scores of each group Table 3 Thickness scores of each group Experiment 3 Effects of different drug doses on psoriasis Experimental group settings: Example 1: A TSLP expression level regulator, which is a TSLP inhibitor. The inhibitor is composed of proguanil and Bacillus subtilis, the mass ratio of the two is 20 mg: 1 g, and the time interval between the use of the two is 2 hours.

[0097] Example 2: A TSLP expression level regulator, which is a TSLP inhibitor. The inhibitor is composed of proguanil and Bacillus subtilis, the mass ratio of the two is 50 mg: 1 g, and the time interval between the use of the two is 2 hours.

[0098] Example 3: A TSLP expression level regulator, which is a TSLP inhibitor. The inhibitor is composed of proguanil and Bacillus subtilis, the mass ratio of the two is 20 mg:1.5 g, and the time interval between the use of the two is 2 hours.

[0099] Control 1: A TSLP expression level regulator, which is a TSLP inhibitor. The inhibitor is composed of proguanil and Bacillus subtilis, the mass ratio of the two is 10 mg: 1 g, and the time interval between the use of the two is 2 hours.

[0100] Control 2: A TSLP expression level regulator, which is a TSLP inhibitor. The inhibitor is composed of proguanil and Bacillus subtilis, with a mass ratio of 50 mg:0.8 g, and the time interval between their use is 2 hours.

[0101] The relevant treatment methods refer to Experiment 2, and the scores of each group are counted. The results are shown in Table 4. The results in Table 4 show that after treatment, Examples 1 to 3 can treat psoriasis. The dosage of proguanil in Control 1 is too low, and the dosage of Bacillus subtilis in Control 2 is too low, and the effect of treating psoriasis is not significant.

[0102] Table 4 Scoring results of each group Experiment 4 Effects of different intervals on psoriasis Example 1: A TSLP expression level regulator, which is a TSLP inhibitor. The inhibitor is composed of proguanil and Bacillus subtilis, the mass ratio of the two is 20 mg: 1 g, and the time interval between the use of the two is 2 hours.

[0103] Example 4: A TSLP expression level regulator, which is a TSLP inhibitor. The inhibitor is composed of proguanil and Bacillus subtilis, the mass ratio of the two is 20 mg: 1 g, and the time interval between the use of the two is 3 hours.

[0104] Example 5: A TSLP expression level regulator, which is a TSLP inhibitor. The inhibitor consists of proguanil and Bacillus subtilis, the mass ratio of the two is 20 mg: 1 g, and the time interval between their use is 4 hours.

[0105] Example 6: A TSLP expression level regulator, which is a TSLP inhibitor. The inhibitor is composed of proguanil and Bacillus subtilis, the mass ratio of the two is 20 mg: 1 g, and the time interval between the use of the two is 0 h.

[0106] Control 3: A TSLP expression level regulator, which is a TSLP inhibitor. The inhibitor consists of proguanil and Bacillus subtilis, with a mass ratio of 50 mg:1 g, and the time interval between their addition is 6 hours.

[0107] The relevant treatment methods refer to Experiment 2, and the scores of each group are counted. The results are shown in Table 5. The results in Table 5 show that the treatment time of Example 1, Example 4 to Example 6 after treatment can effectively treat psoriasis. If the interval time is too long, the effect of treating psoriasis is not significant. When the interval time is 2h~4h, the effect of treating psoriasis is most significant.

[0108] Table 5 Scoring results of each group II It should be noted that when the present invention involves a numerical range, it should be understood that the two endpoints of each numerical range and any value between the two endpoints can be selected. Since the steps and methods used are the same as those in the embodiments, in order to avoid redundancy, the present invention describes a preferred embodiment. Although the preferred embodiments of the present invention have been described, once those skilled in the art know the basic inventive concept, they can make other changes and modifications to these embodiments, and these changes and modifications all fall within the scope of the present invention.

[0109] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. If these modifications and variations of the present invention fall within the scope of the equivalent technology of the present invention, the present invention is also intended to include these modifications and variations.

Claims

1. A TSLP expression level regulator, characterized in that: The TSLP expression level regulator is a TSLP inhibitor, which is composed of proguanil substances and Bacillus subtilis, and the mass ratio of the two is 20mg~50mg:1g~1.5g.

2. The TSLP expression level regulator according to claim 1, characterized in that The proguanil substance is proguanil or a pharmaceutically acceptable salt thereof.

3. The use of the TSLP expression level regulator according to claim 1, characterized in that: The inhibitor is used for preparing medicine for treating psoriasis.

4. The use of the TSLP expression level regulator according to claim 3, characterized in that: The psoriasis is a psoriasis-like skin inflammation caused by abnormal TSLP expression.

5. The use of the TSLP expression level regulator according to claim 3, characterized in that: The inhibitor is used to inhibit the JAK1 / SYK / TRAF6 / NF-κB pathway.

6. The use of the TSLP expression level regulator according to claim 3, characterized in that: The inhibitor is used to reduce the expression levels of IL-17A and IL-17.

7. The use of the TSLP expression level regulator according to claim 3, characterized in that: The inhibitor is used to inhibit the activity of dendritic cells.

8. The TSLP expression level regulator according to claim 3, characterized in that The inhibitor is used to inhibit CD4 + T cells differentiate into Th17 cells.

9. A drug for treating psoriasis, characterized in that: The proguanil substance of claim 1 is used as an active ingredient and is prepared with a pharmaceutically acceptable first auxiliary material to prepare a proguanil substance drug; The Bacillus subtilis is used as an active ingredient and a pharmaceutically acceptable second auxiliary material is added to prepare a Bacillus subtilis medicine; The mass ratio of chlorguanil substances to Bacillus subtilis is 50mg~70mg:1g~1.5g, calculated based on the mass of chlorguanil substances in chlorguanil drugs and the mass ratio of Bacillus subtilis in Bacillus subtilis drugs.

10. The drug for treating psoriasis according to claim 9, characterized in that: The medicine is an oral preparation.

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