Use of micheliolide and formulation containing same

Abstract

The present invention relates to use of a micheliolide (MCL) or a pharmaceutically acceptable salt thereof and a formulation containing same in the preparation of a drug for treating atopic dermatitis or psoriasis. The micheliolide of the present invention can significantly improve related symptoms of psoriasis mice and has a significant therapeutic effect on psoriasis. After transdermal administration, the drug can exhibit good tissue stability and biocompatibility, thereby improving the bioavailability and efficacy, and effectively resisting psoriasis symptoms such as erythema, scaling, and thickening induced by IMQ.

Description

Use of micheliolide and preparation containing the same

[0001] This application claims the priority of the prior application filed by the applicant on July 5, 2024 with the China National Intellectual Property Office, with the patent application number 202410901812.7, and the invention name of "Use of micheliolide and drug-loaded hydrogel preparation containing the same". The content of the above-mentioned prior application is incorporated herein by reference in its entirety. TECHNICAL FIELD

[0002] The present application belongs to the field of pharmaceutical preparations, and relates to the use of micheliolide and a preparation containing the same in the preparation of a medicament for treating atopic dermatitis or psoriasis. BACKGROUND

[0003] Atopic dermatitis is a chronic, recurrent, inflammatory skin disease related to allergic constitution, and its etiology is complex, including genetic, environmental, immune and other factors. The symptoms of atopic dermatitis include skin itching, redness, dryness, scaling, etc., and even blisters, erosion and exudation in severe cases. Due to the complex etiology of atopic dermatitis, the pathogenic inducement cannot be completely eliminated, which makes it prone to recurrence and leads to incurable. At present, for mild atopic dermatitis patients, calcineurin inhibitors, glucocorticoids and other drugs can be used externally; for moderate patients, after the skin lesions are controlled by external drugs, combined drug therapy can be used for active maintenance treatment; for severe patients, systemic immunosuppressive agents, biological agents and phototherapy are considered. In the above treatment methods, both external and systemic treatments have the limitations of narrow applicable population, low cure rate, high price, incomplete clinical data, etc., and long-term use can cause serious adverse reactions such as telangiectasia, ecchymosis, skin atrophy, acne, etc.

[0004] Psoriasis is a chronic immune-mediated inflammatory skin disease that affects about 2-3% of the global population. The pathogenesis of psoriasis is still being explored, and research has confirmed that it involves complex interactions between genetic, immune and environmental factors. In the treatment of mild psoriasis, external corticosteroids, vitamin D3 derivatives, calcineurin inhibitors and keratolytic agents are commonly used, which play a role by regulating gene transcription, but long-term use can cause side effects such as skin atrophy, itching and telangiectasia.

[0005] Micheliolide (MCL) is a natural product of sesquiterpene lactone, mainly distributed in the root bark of Magnoliaceae plants, and there is little research on its activity and preparation development. Therefore, it is necessary to study its drug activity and preparation in order to develop the treatment of skin diseases that are refractory and have serious side effects after chemotherapy. SUMMARY

[0006] In order to improve the prior art, the purpose of the present application is to provide a use of michelalactone and a preparation containing the same in the preparation of a medicament for treating atopic dermatitis or psoriasis.

[0007] The purpose of the present application is achieved by the following technical solutions:

[0008] A use of michelalactone (MCL) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating atopic dermatitis or psoriasis, the michelalactone being shown as formula 1 below:

[0009] According to an embodiment of the present application, the michelalactone or the pharmaceutically acceptable salt thereof is used as a single component or in a composition with other pharmaceutically acceptable components to prepare a medicament for treating atopic dermatitis or psoriasis.

[0010] According to an embodiment of the present application, the pharmaceutically acceptable salt is a sodium salt, a potassium salt, a calcium salt, or an ammonium salt, etc.

[0011] According to an embodiment of the present application, the other pharmaceutically acceptable components can be a drug having no antagonistic effect with the michelalactone, or any one or more excipients allowed in pharmacy.

[0012] According to an embodiment of the present application, the pharmaceutical dosage form is a capsule, a tablet, an oral preparation, an injection, a spray, a gel, an ointment, a cream, or a patch, etc.

[0013] According to an embodiment of the present application, the gel is a hydrogel preparation.

[0014] According to an embodiment of the present application, the administration of the medicament is by injection, oral administration, parenteral administration, inhalation spray, or transdermal administration.

[0015] The present application also provides a drug-loaded hydrogel preparation, comprising a hydrogel matrix, a sesquiterpene lactone compound or a pharmaceutically acceptable salt thereof, a cyclodextrin or a derivative thereof, and water, wherein the content of the sesquiterpene lactone or the pharmaceutically acceptable salt thereof is 0.5-5 mg / mL, preferably 1-5 mg / mL, such as 0.5 mg / mL, 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, or 5 mg / mL, of the drug-loaded hydrogel preparation; the concentration of the hydrogel matrix is 1%-10% (w / v), such as 1% (w / v), 2% (w / v), 3% (w / v), 4% (w / v), 5% (w / v), 6% (w / v), 7% (w / v), 8% (w / v), 9% (w / v), or 10% (w / v), preferably 2% (w / v); and the molar ratio of the sesquiterpene lactone compound or the pharmaceutically acceptable salt thereof to the cyclodextrin or the derivative thereof is 1:1-1:15, such as 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, or 1:15, preferably 1:10.

[0016] According to an embodiment of the present application, in the drug-loaded hydrogel preparation, the hydrogel matrix is loaded with the sesquiterpene lactone compound included by the cyclodextrin or the derivative thereof.

[0017] According to an embodiment of the present application, the hydrogel matrix is a polysaccharide natural high-molecular material, including starch, cellulose, trehalose, hyaluronic acid, chitosan, or a modified product thereof, preferably a hyaluronic acid modified product, and the average molecular weight of the hyaluronic acid is 10-100 million Daltons, preferably 10-50 million Daltons, and more preferably 10-20 million Daltons.

[0018] According to an embodiment of the present application, the preparation method of the hyaluronic acid modified product is as follows: preparing a hyaluronic acid oxidation product, and obtaining the product after cross-linking reaction.

[0019] According to an embodiment of the present application, the sesquiterpene lactone compound is selected from Micheliae lactone (MCL) shown in formula I, the cyclodextrin is β-cyclodextrin, and the derivative of the β-cyclodextrin is hydroxypropyl-β-cyclodextrin.

[0020] According to an embodiment of the present application, the drug-loaded hydrogel preparation is for transdermal administration.

[0021] The present application also provides a preparation method of a drug-loaded hydrogel, comprising the following steps: preparing a compound shown in formula I into a MCL / HP-β-CD inclusion compound, and mixing the compound with a hydrogel matrix.

[0022] According to the embodiment of the present application, in the preparation method, the hydrogel matrix is selected from hyaluronic acid modifiers, which are obtained by preparing hyaluronic acid oxidation products and then performing cross-linking reaction.

[0023] According to the embodiment of the present application, the method specifically comprises the following steps:

[0024] (1) Preparation of hydrogel matrix: preparing hyaluronic acid oxidation products, performing cross-linking reaction to obtain hyaluronic acid modifiers;

[0025] (2) Preparation of MCL / HP-β-CD inclusion complex;

[0026] (3) Preparation of MCL@HA drug-loaded hydrogel.

[0027] According to the embodiment of the present application, in the preparation method, in step (1), hyaluronic acid (HA) is dissolved in water, an oxidizing agent is added, a terminating agent is added to obtain an oxidation product; a cross-linking agent is added to perform reaction to obtain a gel block.

[0028] According to the embodiment of the present application, in the preparation method, in step (1), the oxidation reaction is performed in the dark, and the oxidation product is purified by dialysis, freeze-dried, dissolved and then subjected to cross-linking reaction.

[0029] According to the embodiment of the present application, in the preparation method, in step (1), the oxidizing agent is selected from NaIO4, n(HA):n(NaIO4) is 1:1, the terminating agent is selected from ethylene glycol; and the cross-linking agent is selected from adipic acid dihydrazide, n(HA):n(ADH) is 1:1.

[0030] According to the embodiment of the present application, in the preparation method, in step (1), the obtained gel block is ground to obtain a powder, which is dissolved in water under water bath and cooled to obtain the hydrogel matrix.

[0031] According to the embodiment of the present application, in the preparation method, in step (1), the concentration of HA in the hydrogel is 1-10% (w / v).

[0032] According to the embodiment of the present application, in the preparation method, in step (2), HP-β-CD is precisely weighed and placed in a marver, MCL is added, and the inclusion complex is formed by grinding, PBS solution is added to dissolve and transfer to a clean EP tube, and PBS solution is added to rinse the marver, and then combined to obtain MCL / HP-B-CD inclusion complex solution.

[0033] According to the embodiment of the present application, in the preparation method, in step (2), the molar ratio of MCL to HP-B-CD is 1:1-1:15.

[0034] According to an embodiment of the present application, in the preparation method, the MCL@HA drug-loaded hydrogel is obtained by mixing and oscillating the hydrogel matrix of step (1) with the MCL / HP-β-CD inclusion complex solution of step (2).

[0035] The present application also provides a use of the drug-loaded hydrogel preparation in the preparation of a medicament for treating specific dermatitis or psoriasis.

[0036] According to an embodiment of the present application, in the use, the symptoms of psoriasis such as parakeratosis, acanthosis and extension of dermal papilla are significantly improved.

[0037] The present application also provides a forchlorfenicol ointment, comprising forchlorfenicol or a pharmaceutically acceptable salt thereof and an ointment base, wherein the content of forchlorfenicol or a pharmaceutically acceptable salt thereof is 0.01-10wt%, and the content of the ointment base is 90-99.99wt%. Preferably, the content of forchlorfenicol or a pharmaceutically acceptable salt thereof is 0.01-9wt%, 0.03-9wt%, 0.05-9wt%, 0.07-8wt%, 0.09-8wt%, 0.1-7wt%, 0.15-6wt%, 0.2-8wt%, 0.25-5wt%, 0.5-5wt%, specifically, 0.25wt%, 0.5wt%, 0.6wt%, 0.7wt%, 0.8wt%, 0.9wt%, 1.0wt%, 1.1wt%, 1.2wt%, 1.3wt%, 1.4wt%, 1.5wt%, 1.6wt%, 1.7wt%, 1.8wt%, 1.9wt%, 2.0wt%, 2.1wt%, 2.2wt%, 2.3wt%, 2.4wt%, 2.5wt%, 2.6wt%, 2.7wt%, 2.8wt%, 2.9wt%, 3.0wt%, 3.1wt%, 3.2wt%, 3.3wt%, 3.4wt%, 3.5wt%, 3.6wt%, 3.7wt%, 3.8wt%, 3.9wt%, 4.0wt%, 4.1wt%, 4.2wt%, 4.3wt%, 4.4wt%, 4.5wt%, 4.6wt%, 4.7wt%, 4.8wt%, 4.9wt%, 5.0wt%, 5.1wt%, 5.2wt%, 5.3wt%, 5.4wt%, 5.5wt%, 5.6wt%, 5.7wt%, 5.8wt%, 5.9wt%, 6.0wt%, 6.5wt%, 7.0wt%, 7.5wt%, 8.0wt%, 8.5wt%, 9.0wt%, 9.5wt%, 10.0wt%. Preferably, the content of the ointment base is 92-99.5wt%, 95-99wt%, specifically, 99.75wt%, 99.5wt%, 99wt%.

[0038] According to an embodiment of the present application, the ointment base is selected from one or two of the following: white petrolatum or liquid paraffin.

[0039] According to an embodiment of the present application, the ointment base can further comprise glyceryl monostearate, stearic acid, a preservative, a suspending agent or an anti-settling agent, etc.

[0040] According to an embodiment of the present application, the preservative in the ointment base is selected from one or more of the following: methylparaben, ethylparaben, propylparaben, phenoxyethanol.

[0041] According to an embodiment of the present application, the suspending agent in the ointment base is microcrystalline cellulose; and the anti-settling agent is silicon dioxide.

[0042] According to an embodiment of the present application, in the ointment base, the content of michelia linaloeifera lactone or a pharmaceutically acceptable salt thereof is 0.25-5 wt%, the content of white petrolatum is 50 wt%, and the content of liquid paraffin is 45-49.75 wt%.

[0043] According to an embodiment of the present application, in the ointment base, the content of michelia linaloeifera lactone or a pharmaceutically acceptable salt thereof is 1-5 wt%, the content of white petrolatum is 50-65 wt%, the content of liquid paraffin is 20-30 wt%, the content of glyceryl monostearate is 7-10 wt%, the content of stearic acid is 4-6 wt%, the content of preservative is 0.1-0.5 wt%, the content of suspending agent is 0-2.5 wt%, and the content of anti-settling agent is 0-2.8 wt%.

[0044] The present application also provides a preparation method of the ointment of michelia linaloeifera lactone (MCL), which specifically comprises the following steps:

[0045] 1) Material pretreatment: crush MCL through a 200-mesh sieve, and weigh the ointment oil phase base for standby;

[0046] 2) Main drug mixing: add the MCL fine powder into one ointment oil phase base, stir and grind to ensure uniform dispersion of the drug; then add another ointment oil phase base, and continue to stir and grind;

[0047] 3) Homogenization treatment;

[0048] 4) Filling and cooling.

[0049] According to an embodiment of the present application, in the preparation method, in step 1), the main drug MCL needs to be crushed through a 200-mesh sieve, and the white petrolatum and liquid paraffin are accurately weighed according to the prescription amount for standby.

[0050] According to the embodiment of the present application, in the preparation method, in step 2), the MCL fine powder is added into the liquid paraffin base in batches, and stirred and ground for 30 minutes to ensure uniform dispersion of the drug. Then white vaseline is added, and stirring and grinding is continued for 20 minutes.

[0051] According to the embodiment of the present application, in the preparation method, in step 3), the particle aggregation phenomenon is eliminated by circulating treatment 2-3 times through a colloid mill or a homogenizer, so that the system reaches a visually uniform state.

[0052] According to the embodiment of the present application, in the preparation method, in step 4), the filling precision needs to be controlled within an error range of ±2% to avoid stratification.

[0053] The present application also provides another preparation method of the Michelia Conferta Lactone (MCL) ointment, which specifically comprises the following steps:

[0054] 1) Raw material pretreatment: including main drug and oil phase base treatment;

[0055] 2) Oil phase preparation;

[0056] 3) Main drug addition and dispersion;

[0057] 4) Suspending agent dispersion;

[0058] 5) Cooling and homogenization;

[0059] 6) Filling.

[0060] According to the embodiment of the present application, in the preparation method, step 1) comprises:

[0061] Main drug treatment: MCL is passed through a 200-mesh sieve for standby;

[0062] Oil phase base treatment: white vaseline, liquid paraffin, glycerol monostearate, and stearic acid are sieved respectively to remove impurities.

[0063] According to the embodiment of the present application, in the preparation method, step 2) comprises:

[0064] White vaseline and liquid paraffin are added into a stainless steel container, and heated to 70-75°C in a water bath, and stirred until completely melted;

[0065] Glycerol monostearate and stearic acid are added, the temperature is kept at 70°C, and stirring is continued for 20 minutes until uniform mixing.

[0066] According to the embodiment of the present application, in the preparation method, step 3) comprises:

[0067] The MCL powder is slowly added into the melted oil phase, the temperature is kept at 40-50°C, and stirring is continued at 500-800 rpm for 30 minutes to ensure no particles.

[0068] Add bacteriostatic agent, stir for 10 minutes to dissolve.

[0069] According to the embodiment of the present application, in the preparation method, step 4) comprises: passing the suspending agent through a 120-mesh sieve, slowly sprinkling into the paste, and stirring at 300 rpm for 15 minutes to avoid agglomeration.

[0070] According to the embodiment of the present application, in the preparation method, step 5) comprises: stopping heating, naturally cooling to below 40°C, and transferring to a homogenizer (2000-3000 rpm, homogenizing for 5 minutes) to eliminate bubbles and particles.

[0071] According to the embodiment of the present application, in the preparation method, step 6) comprises: filling the paste into an aluminum tube or a plastic box while the paste is not solidified (35-40°C), and sealing and curing at room temperature.

[0072] The present application also provides use of the forchlorfenuron ointment in preparation of a medicament for treating atopic dermatitis or psoriasis.

[0073] The present application has the following beneficial effects:

[0074] The present application provides use of forchlorfenuron and its preparation in preparation of a medicament for effectively treating atopic dermatitis or psoriasis.

[0075] The forchlorfenuron of the present application can significantly improve the related symptoms of psoriasis mice, has a significant therapeutic effect on psoriasis, after transdermal administration, can play a good tissue stability and biocompatibility of the drug, improve the bioavailability and curative effect, effectively resist the erythema, scales, thickening and other psoriasis symptoms induced by IMQ. BRIEF DESCRIPTION OF DRAWINGS

[0076] Figure 1 MCL can reduce MC903-induced skin itching.

[0077] Figure 2 MCL can reduce the symptoms of atopic dermatitis.

[0078] Figure 3 MCL can reduce MC903-induced skin thickening.

[0079] Figure 4 MCL can reduce MC903-induced skin inflammation.

[0080] Figure 5 MCL can reduce the spleen index of mice.

[0081] Figure 6 MCL can reduce the number of mast cells in skin tissue.

[0082] Figure 7 Flow chart of paraffin section staining.

[0083] Figure 8 Appearance of hydrogel matrix with different HA proportions.

[0084] Figure 9 shows the appearance of MCL / HP-β-CD with different inclusion ratios and the appearance of the initial MCL.

[0085] Figure 10 shows the changes in the back symptoms of the mice in each group over time.

[0086] Figure 11 shows the back erythema, scaling, thickening, and PASI score of the mice in each group.

[0087] Figure 12 shows the changes in the ear symptoms of the mice in each group over time.

[0088] Figure 13 shows the ear erythema, scaling, thickening, and PASI score of the mice in each group.

[0089] Figure 14 shows the Western Blot analysis of the ear tissue.

[0090] Figure 15 shows the relative expression of IL-1β in each group.

[0091] Figure 16 shows the H&E staining analysis of the ear skin tissue of the mice.

[0092] Figure 17 shows the daily back pictures of the treatment of psoriasis in mice by MCL ointment.

[0093] Figure 18 shows the PASI score of the treatment of psoriasis in mice by MCL ointment (skin thickness, scabbing, erythema, and total score, respectively, “###” indicates P<0.001 compared with the blank control; “*” indicates P<0.05 compared with the model control; “**” indicates P<0.01 compared with the model control; “***” indicates P<0.001 compared with the model control. DETAILED DESCRIPTION

[0094] The present application will be further described in conjunction with specific examples. It should be understood that the following examples are merely illustrative and explanatory of the present application and should not be construed as limiting the scope of the present application. Any technology achieved based on the above description of the present application is encompassed within the scope of the present application.

[0095] MCL@HA in the present application: MCL / HP-β-CD-loaded HA drug-loaded hydrogel, i.e., the MCL@HA drug-loaded hydrogel prepared in Preparation Example 2.

[0096] Experimental materials and instruments

[0097] 1. Experimental animals

[0098] For the animal model of specific dermatitis, male C57 / BL6J mice with a body weight of 23-25 g and an age of 6-8 weeks were selected and purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (Beijing, China).

[0099] Animal models for psoriasis were selected from male BALB / c mice aged 6-8 weeks and weighing 20-25 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (Beijing, China). The mice were adaptively fed and the feeding environment during the experiment was maintained at a light cycle of 12 h alternating with day and night, a temperature of 25±2°C, a humidity of 55±5%, and balanced and sufficient standard diet and drinking water. The experimental procedure was approved by the Experimental Animal Center of Hebei University.

[0100] 2. Experimental materials

[0101] The main reagent materials required in the experiment were commercially available, wherein MCL was produced by Tianjin Shandeng Pharmaceutical Technology Co., Ltd.; hydroxypropyl-β-cyclodextrin was produced by Shanghai Yuanye Bio-Technology Co., Ltd.; calcipotriol (MC903) was produced by MedChemExpress; hyaluronic acid (MW: 10-20 million) was produced by Shanghai Macklin Biochemical Technology Co., Ltd.; depilatory cream was produced by Guangzhou Shangdu Cosmetics Co., Ltd.; and 5% imiquimod cream was produced by Sichuan Mingxin Pharmaceutical Co., Ltd.

[0102] 3. Experimental instruments

[0103] The main instruments and equipment required in the experiment were commercially available.

[0104] First part: MCL treatment of atopic dermatitis test

[0105] 1. Establishment of mouse atopic dermatitis model

[0106] Sixteen 6-8-week-old male C57 / BL6J mice were adaptively fed for 3 days, weighed, and all weighed about 25 g. MC903 was applied to the back of the mouse ears, the concentration of MC903 was 100 μM / L, 20 μL was applied to the back of each mouse ear each time, and the application was performed continuously for 7 days. On the 7th day, the mice were randomly divided into a model group and an MCL treatment group, 8 mice in each group. Starting on the 8th day, the model group was normally applied with MC903 in the morning and intraperitoneally injected with solvent in the afternoon; the treatment group was intraperitoneally injected with MCL, normally applied with MC903 in the morning, and intraperitoneally injected with MCL in the afternoon, the concentration of MCL was 10 mg / kg, 100 μL was injected per mouse, and the treatment was continuously performed for 7 days, ending on the 14th day.

[0107] 2. Scratching tracking of mouse itching behavior

[0108] In order to evaluate the treatment effect of MCL, the scratching behavior of each mouse was recorded, and the scratching frequency was statistically analyzed.

[0109] 3. PASI score of mouse ears

[0110] From the first day, the ear skin of each group of mice was observed and photographed before administration every morning, and the mouse ear thickness, dermatitis degree (bleeding / erythema, scale / dryness, edema, abrasion / erosion) was scored according to the Psoriasis Area and Severity Index (PASI) score.

[0111] 4. Material taking

[0112] On the 15th day, the mice were perfused with normal saline, and the mouse ears and spleen were taken for histomorphological observation.

[0113] Experimental results

[0114] 1. MCL can reduce MC903-induced skin itching

[0115] Psoriasis is an extremely itchy inflammatory skin disease. As shown in Figure 1, ear application of MC903 can induce significant scratching behavior in mice. Starting from the 8th day, MCL was administered by intraperitoneal injection for treatment, and compared with the model group, the MCL treatment group can reduce the number of scratching in mice, and relieve the MC903-induced skin itching.

[0116] 2. MCL can reduce MC903-induced skin inflammation

[0117] Compared with the model group, MCL can significantly reduce the symptoms of MC903-induced skin thickening, erythema, scale, etc. (Figure 2), and statistics show that MCL can reduce the skin thickening of mice (Figure 3). Further scoring of the inflammation of the mouse ear skin (Figure 4) found that MCL can reduce the degree of dermatitis induced by MC903.

[0118] 3. MCL can reduce the spleen index of mice

[0119] In order to further verify the effect of MCL on immune regulation of mice, the spleen was taken and the spleen index was calculated. As shown in Figure 5, MCL can reduce the size and index of the spleen of mice with psoriasis.

[0120] 4. MCL can reduce the number of mast cells in the skin

[0121] As shown in Figure 6, MCL can significantly reduce the number of mast cells in the skin, thereby playing a role in inhibiting psoriasis.

[0122] Preparation of drug-loaded hydrogel and its treatment of psoriasis

[0123] I. Establishment of animal model

[0124] Example 1 Establishment of IMQ-induced mouse psoriasis model

[0125] 1. Establishment of mouse psoriasis model on the back

[0126] Take 10 6-8 week old male BALB / c mice, adaptively feed for 3 days, weigh, all about 22g. Randomly divided into 3 groups, of which 2 are blank (Vehicle), 4 are model (IMQ-Model), and 4 are matrix (IMQ+HA). Use a shaver and depilatory cream to remove the hair on the back of the mice 8-9 cm 2 From the next day (Day 0), at 9 am every day, the matrix group is applied with 200 μL HA empty matrix, and the model group is rubbed with an appropriate amount of PBS solution on the back skin with cotton wool; at 5 pm, the matrix and model groups are applied with 62.5 mg / each of 5% imiquimod cream, until the end of Day 6 (Day 6).

[0127] 2. Establishment of mouse psoriasis model

[0128] Take 7 6-8 week old male BALB / c mice, adaptively feed and weigh as in the method of the experimental animals section of the experimental materials and instruments. Randomly divided into 2 groups, of which 4 are model (IMQ-Model) and 3 are treatment (IMQ+MCL@HA). From Day 0, at 9 am every day, apply 62.5 mg / each of 5% imiquimod cream to the left ear of the 7 mice, and at 5 pm apply 200 μL MCL@HA drug-loaded hydrogel to the treatment group. On Day 7 (Day 7), the mice are sacrificed by cervical dislocation, and all the left ears and normal right ears of the treatment group mice are cut off. Each ear is cut into three parts from the outside to the inside, and the middle part is placed in an EP tube containing 4% paraformaldehyde general tissue fixative and stored at 4°C; the two side parts are placed in 2 mL enzyme-free EP tubes and stored at -80°C for later use.

[0129] 3. Mouse back and ear PASI score

[0130] From Day 1, observe the back or ear skin of the mice in each group every morning before administration, and score the symptoms of erythema, scaling, and thickening according to the Psoriasis Area and Severity Index (PASI), with 0-4 points for each symptom from light to severe, and the total score is the sum of the three scores, 0-12 points, and a line graph is drawn for comparison.

[0131] II. Preparation Examples

[0132] Example 2 Preparation of MCL@HA drug-loaded hydrogel

[0133] 1. Preparation of HA-ADH gel block

[0134] Dissolve 2.0 g of hyaluronic acid (MW: 10-20 million) in 200 mL of double distilled water, stir until completely dissolved, add NaIO4 (MW: 213.89) 1.066 g, so that n(HA): n(NaIO4) = 1:1, react at room temperature for 24 h in the dark, add 2 mL of ethylene glycol to stop the reaction to obtain the oxidation product.

[0135] Cut 10-12 cm dialysis bag 5 parts in double distilled water and boil for 10 min, cool at room temperature. Transfer the above product to the pretreated dialysis bag, and place it in a measuring cup containing 2 L of double distilled water for dialysis. Replace the double distilled water at 2-4 h, 6-8 h and 10-14 h after the start of dialysis, respectively, continue dialysis for 2 h after the last liquid change, and end the dialysis when no precipitate is observed.

[0136] Concentrate the purified product to one-tenth of the original volume by evaporation, and freeze-dry overnight. Dissolve the freeze-dried product in a small amount of double distilled water, and add adipic acid dihydrazide (ADH, MW: 174.2) 0.868 g as a crosslinking agent, so that n(HA): n(ADH) = 1:1, react at 4°C overnight. Freeze-dry the final product for 24 h to obtain HA-ADH gel block, grind into powder with a maroon mortar, and freeze for later use.

[0137] 2. Preparation of hydrogel matrix with different HA proportions

[0138] Take clean 10 mL EP tubes 4, labeled 1%, 2%, 4%, 8%, and add 4 mL of pure water respectively. Take HA-ADH gel powder 0.04 g, 0.08 g, 0.16 g, 0.32 g respectively and add to the corresponding EP tubes, mix and stir at room temperature for 30 min, then heat in a 90°C water bath for 1 h to promote dissolution, cool to room temperature, and stand at 4°C overnight to obtain hydrogel matrix with different HA proportions. Transfer them respectively to colorless and transparent sample bottles, seal with lids, tilt and invert, and observe and compare the appearance and flow characteristics of hydrogel matrix with different HA proportions.

[0139] 3. Preparation of MCL / HP-β-CD inclusion complex with different inclusion ratios

[0140] Precisely weigh 1 mg of MCL (MW: 248.32) 3 times and set aside. Precisely weigh HP-β-CD (MW: 1541.54) 6.2 mg, 0.06208 g, respectively, and place in a maroon mortar. Add 1 mg of MCL and grind for 40 min to form an inclusion complex. Add 0.5 mL of PBS solution to dissolve and transfer to a clean EP tube. Add 0.5 mL of PBS solution to rinse the mortar, and combine to obtain MCL / HP-B-CD inclusion complex solution with a drug content of 1 mg / mL and a molar ratio of 1:1, 1:10, respectively.

[0141] 1 mg MCL was placed in a clean EP tube, 1 mL PBS solution was added to obtain the original MCL drug solution. The three were transferred to a colorless transparent sample bottle, and the solubility of MCL or MCL / HP-β-CD inclusion complex was observed and compared. The inclusion ratio corresponding to the clearest solution was selected as the ratio of MCL / HP-β-CD inclusion complex mother liquor (4 mL, drug content 2 mg / mL) used for subsequent preparation of drug-loaded hydrogel.

[0142] 4. Preparation of MCL@HA drug-loaded hydrogel

[0143] After evaluating the appearance and flowability of hydrogel matrices with different HA ratios, a matrix with good permeability, coating properties, stability, and aesthetic appearance was selected as the carrier for MCL / HP-β-CD inclusion complex.

[0144] According to the preparation method of hydrogel matrix with different HA ratios in Example 2, Part 2, 4 mL of HA hydrogel with double the above ratio was prepared, mixed with 4 mL of MCL / HP-β-CD mother liquor, vortexed, and 8 mL of drug-loaded hydrogel with a drug content of 1 mg / mL was obtained. Store at 4°C for later use.

[0145] III. Experiment and Effect

[0146] Example 3 Western Blot Analysis Experiment

[0147] 1. Protein extraction

[0148] Prepare the tissue lysis solution (high-efficiency RIPA lysis solution 3 mL + PMSF protease inhibitor 30 μL + phosphatase inhibitor 30 μL + Protease Inhibitor Cocktail 30 μL). Take the mouse ear samples from the -80°C refrigerator and place them on ice, add 200 μL of lysis solution to each, and homogenize with a handheld high-speed homogenizer on ice to mix the tissue and lysis solution thoroughly. Add another 100 μL of lysis solution, vortex for 30 s, and lyse on ice for 30 min, then centrifuge for 20 min (4°C, 12000 rpm), take 5 μL of supernatant in a clean EP tube, add 20 μL of ultrapure water to dilute, and store on ice.

[0149] 2. Protein concentration determination

[0150] Diluted protein samples were centrifuged for 15 s, 10 μL per well was added to a 96-well plate, and BSA protein standard solution with gradient concentrations was added, 2 replicates for each sample. BCA protein concentration assay reagent (4.4 mL of solution A and 88 μL of solution B) was prepared, and 100 μL was added to each replicate. The mixture was reacted in a 37°C oven for 30 min in the dark, and then the absorbance of each well was detected by a microplate reader. A standard curve was drawn and the protein loading amount was calculated. The calculated results were added to a clean EP tube, and the tube was vortexed and centrifuged for 15 s. The mixture was boiled for 5 min, cooled to room temperature, and stored at -20°C for standby.

[0151] 3. Gel preparation

[0152] Clean and dry glass plates were aligned on a gel holder and clamped in a gel tank. The gap between the two plates was filled with water to check for leaks for 20 min. After passing the test, the upper and lower gels were prepared according to the instructions of the PAGE gel rapid preparation kit, and then added to the gap between the two plates. The comb was inserted, and the gels were allowed to solidify for 40 min.

[0153] 4. Electrophoretic separation

[0154] The glass plates were removed and installed in the electrophoresis tank, which was then filled with electrophoresis buffer. The comb was removed, and the comb holes were arranged. Then, 5 μL of Marker and 10 μL of sample were added, respectively, and 10 μL of 1×loading was added to the comb holes at both ends to prevent band skewing.

[0155] The cover plate of the electrophoresis apparatus was covered, and the positive and negative electrodes were connected. The electrophoresis was started. The initial voltage was set to 90 V, and after 30 min, it was increased to 120 V. When the bromophenol blue moved to the lower edge of the gel, the electrophoresis was stopped.

[0156] 5. Membrane transfer

[0157] The upper part of the membrane transfer clamp was covered with an asbestos net and filter paper, and then soaked in the transfer solution cooled at 4°C. The glass plates were removed from the electrophoresis tank and carefully pried open. The upper gel and lower bromophenol blue were cut off, and the gel was cut along the edge of the Marker and placed on the upper part of the black clamp. A PVDF membrane of the same size was cut, activated in methanol for 15 s, and then soaked in the transfer solution and placed on the white clamp. The bubbles were squeezed out with a roller, and the clamps were clamped together and placed in a transfer tank containing an ice box. The cover plate was covered, and the positive and negative electrodes were connected. The membrane was transferred at 250 mA on ice for 2 h.

[0158] 6. Blocking and antibody incubation

[0159] After the transfer was completed, the PVDF membrane was clamped with tweezers into an incubation box containing an appropriate amount of skim milk blocking solution (2.5 g of skim milk powder + 50 mL of TBST), and placed on a shaker with the rotation speed adjusted to 100 rpm. Blocking was performed for 2 h. After completion, the blocking solution was discarded, and an appropriate amount of TBST was added to wash the membrane for 10 min, and the process was repeated three times.

[0160] According to the molecular weight of the protein to be tested, the required PVDF membrane is cut and placed in an incubation box containing the corresponding primary antibody. Incubate overnight at 4°C, remove the next morning, and incubate at room temperature for 2 hours on a shaker. Recover the primary antibody. Add an appropriate amount of TBST to wash the membrane for 10 minutes, repeat three times, then add the corresponding secondary antibody and incubate for 1 hour. After incubation, add an appropriate amount of TBST to wash the membrane for 10 minutes, repeat three times.

[0161] 7. Development

[0162] Carefully clamp the PVDF membrane with tweezers, and gently dab the remaining TBST on the paper. Soak the membrane evenly with freshly prepared ECL chemiluminescence solution (A solution: B solution = 1:1), and develop it on a chemiluminescence gel imaging system. β-actin is the internal reference antibody.

[0163] Table 1 Antibodies involved in Western Blot Analysis

[0164] Example 4 Real-time fluorescent quantitative PCR (qPCR) detection

[0165] 1. RNA extraction

[0166] Take the mouse ear samples from the -80°C freezer and place them on ice. Add 200 μL of RNAiso Plus to each sample, and homogenize them using a handheld high-speed homogenizer. Add an additional 300 μL of RNAiso Plus, and let it stand at room temperature for 5 minutes. Add 100 μL of chloroform, and quickly shake for 15 seconds. Let it stand at room temperature for 2 minutes, then centrifuge for 15 minutes at 4°C and 12,000 rpm. Transfer the supernatant to a clean enzyme-free EP tube, and add an equal volume of isopropanol. Mix gently, then let it stand at room temperature for 10 minutes to allow the solution to separate. Centrifuge for 10 minutes at 4°C and 12,000 rpm, discard the supernatant, and add 1 mL of 75% ethanol solution. Centrifuge for 5 minutes at 4°C and 7,500 rpm, discard the supernatant, and invert the EP tube on a clean paper for 10 minutes. Dry the ethanol, and add 10 μL of DEPC water to dissolve the RNA precipitate. Centrifuge for 15 seconds, and store for later use.

[0167] 2. RNA concentration determination, reverse transcription, and PCR amplification

[0168] Take 1.5 μL of the RNA sample and measure its concentration using a ultramicro spectrophotometer. Dilute the sample to control the concentration at 1000-3000 ng / μL, A 260 / A 280 and A 260 / A 230The value is less than 2. The volume of RNA and DEPC water required for reverse transcription is calculated according to the law, added to a 100 μL PCR tube, mixed with 4 μL of All-In-One 5X RT MasterMix, centrifuged for 10 s, placed in a PCR amplifier, the volume was set to 20 μL, 37°C, 15 min, 60°C, 10 min, 95°C, 3 min gradient operation, and the cDNA obtained after the end was stored at 4°C for standby.

[0169] 3. qPCR detection

[0170] The amplified cDNA was diluted 8 times, three replicate wells were set for each sample, 2X qPCR MasterMix 10 μL, 0.5 μL of forward and reverse primers, 7 μL of DEPC water, and 2 μL of cDNA dilution were added to each well of the PCR 8-row tube microplate, and then an enzyme-free film was used to cover it, avoid light, centrifuge the microplate in the centrifuge for 60 s, and place it in a fluorescence quantitative PCR instrument for detection. GAPDH is the internal reference gene.

[0171] Table 2 Primers involved in qPCR experiment

[0172] Example 5 H&E staining

[0173] The fixed mouse ear tissue was taken out, trimmed, washed with paraformaldehyde solution, dehydrated in different concentrations of ethanol solution, immersed in a mixture of paraffin and stearic acid to make the tissue transparent, then treated with wax, the tissue was taken out and placed in an embedding tank, and after embedding, it was quickly placed on a cold table for solidification. The paraffin sectioning machine was set to a thickness of 5 μm, and after the water was developed, the glass slide was vertically lifted, tilted and placed to dry the water, and placed in a 37°C oven overnight. The steps of staining and making slides are shown in Figure 7.

[0174] Example 6 Appearance and flowability evaluation of different HA ratio hydrogel matrices

[0175] As shown in Figure 8, the four ratio matrices were all colorless and transparent, the 1% HA-containing matrix had strong flowability and little wall sticking; the 2% HA-containing matrix had strong flowability, certain viscosity and a little wall sticking; the 4% HA-containing matrix had obvious wall sticking and viscous texture; the 8% HA-containing matrix could hardly flow spontaneously and was gel-like.

[0176] Considering the actual administration, in order to avoid uneven coating and poor skin surface stability, 2% HA matrix was selected as the drug carrier.

[0177] Example 7 MCL solubility evaluation

[0178] As shown in Figure 9, MCL itself is poorly soluble, and the solid quickly settles in the solvent; the MCL / HP-β-CD 1:1 inclusion complex is in suspension, indicating that MCL cannot be effectively dissolved to exert its efficacy at this inclusion ratio; the MCL / HP-β-CD 1:10 inclusion complex is clear and transparent, indicating that the inclusion complex is formed and the solubility of MCL is good, which helps to improve its bioavailability. Therefore, the MCL / HP-β-CD 1:10 inclusion complex mother liquor is prepared.

[0179] Example 8 Mouse back experiment excludes the therapeutic effect of HA empty matrix

[0180] IMQ was continuously applied to the backs of the model group and matrix group mice for 6 days to establish a psoriasis model, as shown in Figure 10, compared with the blank group, IMQ caused the mice to have obvious symptoms of erythema, scaling and thickening, indicating that the model was successfully established. The symptoms of each group were scored and the total PASI score was calculated, and a fold line graph was drawn using GraphPad Prism 10 and subjected to two-way ANOVA and multiple comparisons (Figure 11), it was found that the scores of the model group and the matrix group tended to be consistent over time, and there was no significant difference, indicating that the HA empty matrix had no therapeutic effect, and it was feasible to use it as a drug carrier.

[0181] Example 9 Mouse ear PASI score verifies the pharmacological effect of MCL

[0182] IMQ was continuously applied to the left ears of the model group and treatment group mice for 7 days, and the treatment group mice were given 200 μL of drug-loaded hydrogel containing 1 mg / mL of drug and 2% HA to counteract the symptoms of psoriasis, as shown in Figure 12, the symptoms of the treatment group were significantly relieved, and the score reached a low point on the 5th day, which was manifested as scaling and shedding, which may be related to the behavior and metabolic cycle of the mice. By scoring the symptoms of the two groups and calculating the total PASI score, a fold line graph was drawn and the data was analyzed (Figure 13), it was found that the scores of the treatment group and the model group had significant differences on the 5th to 7th days (p<0.0001). It is proved that MCL has a more obvious therapeutic effect on the appearance evaluation of psoriasis in mice.

[0183] Example 10 Effect of MCL on protein expression at psoriasis site

[0184] Western Blot analysis was performed on samples incubated with p-p65 and IL-1β antibodies to determine their expression, which can analyze the effect of MCL on protein expression at psoriasis site. As shown in Figure 14, the model group significantly up-regulated the expression of IL-1β, while the treatment group had no significant difference from the normal condition; the expression of p-p65 in the model group also showed an up-regulation trend compared with the blank group and the treatment group. It is proved that MCL can down-regulate the protein expression of IL-1β and p-p65 at psoriasis site, making it close to the normal level, and exerting a therapeutic effect.

[0185] Example 11 MCL improves psoriasis symptoms by inhibiting IL-1β expression

[0186] To further verify the effect of MCL on IL-1β, primers were designed for qPCR detection, with GAPDH as the internal reference. As shown in Figure 15, compared with the blank group and the treatment group, the relative gene expression of IL-1β in the model group increased significantly, and there was a significant difference (p<0.05), while there was no significant difference in the relative gene expression between the blank group and the treatment group. It is proved that MCL plays a therapeutic role by significantly down-regulating the expression of IL-1β in psoriasis.

[0187] Example 12 Histopathological morphology analysis

[0188] The pathological analysis of the paraffin section of the mouse ear tissue was carried out by H&E staining, as shown in Figure 16, compared with the blank group, the model group showed local hyperkeratosis and parakeratosis, and the phenomenon of significantly thickening of the prickle layer, and the skin protrusion could be observed to extend to the bottom, proving that the psoriasis model was successfully established. After MCL@HA drug intervention, the epidermal parakeratosis, prickle layer thickening and skin protrusion extension were significantly improved, verifying that MCL has a pharmacological effect of inhibiting the occurrence of psoriasis symptoms.

[0189] Third part: Therapeutic effect of Michelia champaca lactone (MCL) preparation (ointment) on psoriasis

[0190] I. Preparation Example:

[0191] Example 1: MCL ointment was prepared by using F-1, F-2 and F-3 formula.

[0192] F-1

[0193] F-2

[0194] F-3

[0195] The preparation method of MCL ointment is as follows:

[0196] 1. Material pretreatment

[0197] The main drug MCL needs to be crushed through a 200-mesh sieve. White vaseline and liquid paraffin are accurately weighed according to the prescription amount for standby use.

[0198] 2. Main drug mixing

[0199] Add MCL fine powder to the liquid paraffin base in batches, stir and grind for 30 minutes to ensure uniform dispersion of the drug. Then add white vaseline and continue to stir and grind for 20 minutes.

[0200] 3. Homogenization treatment

[0201] Eliminate the phenomenon of particle aggregation by circulating treatment 2-3 times with colloid mill or homogenizer, so that the system reaches the state of visible uniformity.

[0202] 4. Filling and cooling

[0203] Filling into ointment tube, the filling accuracy needs to be controlled within ±2% error range to avoid stratification.

[0204] II. Experiment of treating psoriasis with Michelia Love Latone (MCL) ointment

[0205] 1. Experimental materials

[0206] 1.1 Test samples

[0207] 1.1.1 Drug name 1: 1% MCL ointment (formula F-1);

[0208] Characteristics: milky white ointment; specifications: 10g;

[0209] Drug name 2: 0.5% MCL ointment (formula F-2);

[0210] Characteristics: milky white ointment; specifications: 10g;

[0211] Drug name 3: 0.25% MCL ointment (formula F-3);

[0212] Characteristics: milky white ointment; specifications: 10g.

[0213] 1.1.2 Positive drug: Compound Dexamethasone Acetate Cream;

[0214] Production batch number: 05523009; specifications: 20g;

[0215] Production unit: Hubei Renfu Chengtian Pharmaceutical Co., Ltd.

[0216] 1.1.3 Modeling drug: Imiquimod Cream;

[0217] Production batch number: 24104001; specifications: 3g;

[0218] Production unit: Sichuan Mingxin Pharmaceutical Co., Ltd.

[0219] 1.3 Experimental animals and feeding conditions

[0220] Balb / C mice, 6-8 weeks, male, purchased from Beijing Huafukang Experimental Animal Technology Co., Ltd. The test animals were raised in the sterile independent air supply IVC cage in the Experimental Animal Center of the Institute of Radiation Medicine, Chinese Academy of Medical Sciences, 4-5 animals per cage. The bedding was high-pressure sterilized mouse special bedding, and the animals were fed with sterilized feed specially prepared for mice and free access to pure water. The temperature in the animal laboratory was maintained at about 25℃, the relative humidity was maintained at 40-70%, and the light was on for 12 hours a day.

[0221] 2 Experimental method and grouping

[0222] 2.1 Experimental method

[0223] After 3 days of adaptive feeding, the mice were randomly divided into 6 groups according to their body weight, with 9 mice in each group. On the day of grouping, the skin on the back of the mice was depilated, and the fur was removed with depilatory cream and then wiped clean. The next day, 5% imiquimod cream (about 62.5 mg) was applied to the back of the mice, and after 5-6 hours, the back of the mice was treated, i.e., 1% MCL ointment, 0.5% MCL ointment, 0.25% MCL ointment, and compound dexamethasone acetate cream were applied, once a day for 8 consecutive days. The back of the mice was observed every morning before administration, and the Psoriasis Area and Severity Index (PASI) score was recorded every two days. At the end of the experiment, half of the back skin tissue was quick-frozen and stored at -80℃, and the other half was fixed with tissue fixative for subsequent testing.

[0224] 2.2 Experimental grouping

[0225] (1) Blank control group;

[0226] (2) Model control group;

[0227] (3) Dexamethasone cream

[0228] (4) 1% MCL ointment group;

[0229] (5) 0.5% MCL ointment group;

[0230] (6) 0.25% MCL ointment group;

[0231] Each group had 9 mice, and a total of 54 mice.

[0232] 3 Psoriasis PASI scoring criteria for mice:

[0233] 3.1. Skin thickness:

[0234] 0 points: smooth skin without wrinkles.

[0235] 1 point: slight wrinkles on the skin at the edge of the treated area.

[0236] 2 points: Slight wrinkle on the skin of the drug application area.

[0237] 3 points: Further deepening of the wrinkle on the drug application area.

[0238] 4 points: On the basis of 3 points, the mouse shows weight loss or poor condition.

[0239] 3.2. Crusting (scaling):

[0240] 0 points: Smooth skin without scaling.

[0241] 1 point: Slight scaling on the skin of the drug application area.

[0242] 2 points: The skin of the drug application area is completely covered with scales.

[0243] 3 points: Further deepening of the scaling on the drug application area.

[0244] 4 points: On the basis of 3 points, the mouse shows weight loss or poor condition.

[0245] 3.3. Erythema:

[0246] 0 points: Smooth skin.

[0247] 1 point: Slight redness on the skin of the drug application area.

[0248] 2 points: The skin of the drug application area is completely red.

[0249] 3 points: Further deepening of the redness on the drug application area.

[0250] 4 points: On the basis of 3 points, the mouse shows weight loss or poor condition.

[0251] The total score is the sum of the scores of skin thickness, crusting and erythema.

[0252] 4 Results

[0253] After the second day of modeling, the mouse back appeared scales, and after the third day of modeling, the skin of the mouse back drug application area edge appeared slight wrinkle, the skin appeared slight redness. The model control group showed stable skin thickening, scaling and erythema formation characteristics after the fifth day of modeling, and remained relatively stable within a certain period of time, proving that the mouse psoriasis modeling was successful. Details of the results are shown in Figures 17 and 18.

[0254] 5 Conclusion

[0255] The 0.25%, 0.5% and 1% MCL ointments can significantly improve the symptoms of psoriasis mice and have a significant therapeutic effect on psoriasis.

[0256] Part IV Stability Study of Xiaoshunlactone Ointment

[0257] I. Preparation Example

[0258] Example 2: Preparation of Magnolia Lactone Ointment using F-4, F-5 formula.

[0259] F-4

[0260] F-5

[0261] Preparation method as follows:

[0262] 1) Raw material pretreatment

[0263] · Main drug treatment: MCL passes 200 mesh sieve, ready for use.

[0264] · Oil phase matrix treatment: white vaseline, liquid paraffin, glycerol monostearate, stearic acid are sieved respectively to remove impurities.

[0265] 2) Oil phase preparation

[0266] · Add white vaseline and liquid paraffin to a stainless steel container, heat to 70-75°C in a water bath, and stir until completely melted.

[0267] · Add glycerol monostearate and stearic acid, maintain temperature at 70°C, and stir for 20 minutes until evenly mixed.

[0268] 3) Main drug addition and dispersion

[0269] · Slowly add MCL powder to the melted oil phase, maintain temperature at 40-50°C, and stir at 500-800 rpm for 30 minutes to ensure no particles.

[0270] · Add bacteriostatic agent, stir for 10 minutes to dissolve.

[0271] 4) Suspending agent dispersion

[0272] · Pass the suspending agent through a 120 mesh sieve, slowly sprinkle into the paste, and stir at 300 rpm for 15 minutes to avoid clumping.

[0273] 5) Cooling and homogenization

[0274] · Stop heating, naturally cool to below 40°C, and transfer to a homogenizer (2000-3000 rpm, homogenize for 5 minutes) to eliminate bubbles and particles.

[0275] 6) Filling

[0276] · Fill into aluminum tubes or plastic boxes while the paste is not solidified (35-40°C), seal and solidify at room temperature.

[0277] II. Stability test of Michelia compressa lactone ointment: accelerated stability (40℃, content and properties were investigated)

[0278] Experimental method: the ointment was put into a stability test box (40℃, 75% RH), and the sample was taken out for testing properties and content according to the specified days, and the quality change of the ointment was observed.

[0279] Experimental conclusion: under the accelerated stability test conditions, the properties and content of the sample did not change significantly, and the quality was stable. It is easy to store.

[0280] The above describes the embodiments of the present application. However, the present application is not limited to the above-described embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. Use of micheliolide (MCL) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating atopic dermatitis or psoriasis, characterized in that, The magnolia lactone is shown below as Formula I:

2. Use according to claim 1, characterized in that, The michelia lactone or its pharmaceutically acceptable salt as a single component or with other pharmaceutically acceptable components to prepare a medicine for treating specific dermatitis or psoriasis, wherein the pharmaceutically acceptable salt can be preferably sodium salt, potassium salt, calcium salt or ammonium salt, etc.; the other pharmaceutically acceptable components can be preferably a medicine without antagonism with michelia lactone, or any one or more excipients allowed in pharmacy.

3. Use according to claim 1, characterized in that, The medicine dosage form is capsule, tablet, oral preparation, injection, spray, ointment, cream, gel preparation or patch, etc.

4. Use according to claim 1, characterized in that, The medicine administration mode is injection, oral administration, parenteral administration, inhalation spray or transdermal administration.

5. A drug-loaded hydrogel formulation, characterized in that, The preparation comprises a hydrogel matrix, a sesquiterpene lactone or its pharmaceutically acceptable salt, a cyclodextrin or its derivative and water, wherein the content of the sesquiterpene lactone or its pharmaceutically acceptable salt is 0.5-5 mg / mL, preferably 1-5 mg / mL, such as 0.5 mg / mL, 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, the concentration of the hydrogel matrix is 1%-10% (w / v), such as 1% (w / v), 2% (w / v), 3% (w / v), 4% (w / v), 5% (w / v), 6% (w / v), 7% (w / v), 8% (w / v), 9% (w / v), 10% (w / v), preferably 2% (w / v); the molar ratio of the sesquiterpene lactone or its pharmaceutically acceptable salt to the cyclodextrin and its derivative is 1:1-1:15, such as 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, preferably 1:10, and the hydrogel matrix in the drug-loaded hydrogel preparation is loaded with the sesquiterpene lactone wrapped by the cyclodextrin or its derivative.

6. The preparation according to claim 5, characterized in that The sesquiterpene lactone is selected from the group consisting of magnolialactone having the following formula I: The cyclodextrin is β-cyclodextrin, the β-cyclodextrin derivative is hydroxypropyl-β-cyclodextrin, the hydrogel matrix is hyaluronic acid or its modified product, the average molecular weight of the hyaluronic acid is 10-100 million Dalton, preferably 10-50 million Dalton, more preferably 10-20 million Dalton, and the hyaluronic acid modified product is prepared by the following method: preparing a hyaluronic acid oxidation product, and then performing a cross-linking reaction.

7. A process for the preparation of the drug-loaded hydrogel formulation according to claim 6, characterized by, The preparation method of the drug-loaded hydrogel comprises the following steps: preparing the compound shown in formula I into MCL / HP-B-CD inclusion compound, and mixing with the hydrogel matrix.

8. A forbesia laurina lactone ointment characterized by, The preparation comprises a hydrogel matrix, a sesquiterpene lactone or its pharmaceutically acceptable salt, a cyclodextrin or its derivative and water, wherein the content of the sesquiterpene lactone or its pharmaceutically acceptable salt is 0.5-5 mg / mL, preferably 1-5 mg / mL, such as 0.5 mg / mL, 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, the concentration of the hydrogel matrix is 1%-10% (w / v), such as 1% (w / v), 2% (w / v), 3% (w / v), 4% (w / v), 5% (w / v), 6% (w / v), 7% (w / v), 8% (w / v), 9% (w / v), 10% (w / v), preferably 2% (w / v); the molar ratio of the sesquiterpene lactone or its pharmaceutically acceptable salt to the cyclodextrin and its derivative is 1:1-1:15, such as 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, preferably 1:10, and the hydrogel matrix in the drug-loaded hydrogel preparation is loaded with the sesquiterpene lactone wrapped by the cyclodextrin or its derivative.

9. The methyleugenol ointment according to claim 8, wherein The ointment base is selected from one or two of white petrolatum or liquid paraffin; preferably, the ointment base can further comprise glyceryl monostearate, stearic acid, a preservative, a suspending agent or an anti-settling agent, etc. Preferably, in the ointment base, the preservative is selected from one or more of nipagin, nipagin E, nipagin P or phenoxyethanol; Preferably, in the ointment base, the suspending agent is microcrystalline cellulose; the anti-settling agent is silicon dioxide; Preferably, in the ointment base, the forbesione or pharmaceutically acceptable salt thereof is 1-5 wt%, white petrolatum is 50-65 wt%, liquid paraffin is 20-30 wt%, glyceryl monostearate is 7-10 wt%, stearic acid is 4-6 wt%, the preservative is 0.1-0.5 wt%, the suspending agent is 0-2.5 wt%, and the anti-settling agent is 0-2.8 wt%.

10. Use of the drug-loaded hydrogel preparation of claim 5 or the forbesione ointment of claim 8 in the preparation of a medicament for treating specific dermatitis or psoriasis.

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