Material for treating inflammatory skin diseases, preparation method and application

Abstract

The invention discloses a material for treating inflammatory skin diseases, a preparation method and application, the inflammatory skin diseases comprise psoriasis and atopic dermatitis, and the material comprises cerium substituted molybdenum-based polyoxometallate. The application has the advantages that the CMB has reliable biological safety and excellent active oxygen scavenging capacity, and can effectively inhibit the level of ROS (reactive oxygen species) in cells at lower concentration, so that the inflammation activation of psoriasis or atopic dermatitis is inhibited, and the effect of treating psoriasis or atopic dermatitis is achieved; compared with a water-insoluble inorganic nano material, the CMB material can be dissolved in water, is not easy to retain in local tissues, is more easily metabolized by an organism, and is better in safety.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a material, a preparation method and an application thereof for treating inflammatory skin diseases. Background Art

[0002] Psoriasis, commonly known as psoriasis, is one of the most common chronic recurrent inflammatory skin diseases. It has a long course and is prone to relapse. The clinical manifestations are mainly erythema and scaling. It can occur all over the body, but is more common on the scalp and extensor side of the limbs. It often worsens in winter. The etiology is complex, and the pathogenesis of psoriasis is not very clear. Current studies have found that the onset of psoriasis is related to tumor necrosis factor-α and its related cytokines. A large number of studies have also shown that the occurrence of psoriasis is closely related to the production of reactive oxygen species. At present, it has been reported that natural products or antioxidants can be used to reduce the level of reactive oxygen species in psoriasis for the treatment of psoriasis.

[0003] Atopic dermatitis is a chronic, recurrent, inflammatory skin disease, with recurrent chronic eczematous rashes as the main clinical manifestation, accompanied by significant dryness and itching of the skin. Because patients often have other atopic diseases such as allergic rhinitis and asthma, it is considered a systemic disease. Atopic dermatitis is currently incurable and can seriously affect the quality of life. However, effective and standardized treatment can relieve symptoms and reduce recurrences.

[0004] In recent years, polyoxometalates (POMs) have developed rapidly in the field of biological applications. Polyoxometalates refer to a class of metal oxygen cluster compounds with special properties and structures formed by the aggregation of some transition metal elements. Based on this, many literatures have applied POMs to inflammatory bowel disease, acute kidney injury and other inflammatory-related diseases, and have achieved good therapeutic effects. Molybdenum-based polyoxometalate (POM) nanoclusters are polyoxometalates containing at least molybdenum ions. Under certain reducing or oxidizing conditions, the Mo ions in POMs are in the form of Mo 5+ and Mo 6+ The valence states of these molecules change at any time, and they can react quickly with reactive oxygen species (ROS) to remove ROS in local tissues, thereby alleviating inflammation and tissue damage caused by inflammation. However, a higher concentration is required to effectively inhibit the level of ROS in cells, so the treatment efficiency is not high. Summary of the invention

[0005] The purpose of the present invention is to provide a material, preparation method and application for treating inflammatory skin diseases in view of the deficiencies in the prior art, so as to solve the problem of low treatment efficiency existing in the related art.

[0006] To achieve the above object, the technical solution adopted by the present invention is:

[0007] In a first aspect, a material for treating inflammatory skin diseases is provided, comprising a cerium-substituted molybdenum-based polyoxometalate (CMB);

[0008] Wherein, the inflammatory skin diseases include psoriasis and atopic dermatitis.

[0009] In a second aspect, a preparation method is provided for preparing the material for treating inflammatory skin diseases as described in the first aspect, comprising:

[0010] preparing a mixed solution containing an inorganic cerium salt and a molybdate, wherein the mixed solution is acidic;

[0011] The mixed solution is dried to obtain cerium-substituted molybdenum-based polyoxometalate.

[0012] In some embodiments, the inorganic cerium salt includes cerium chloride, cerium nitrate, and cerium acetate.

[0013] In some embodiments, the molybdate includes sodium molybdate, potassium molybdate, and ammonium molybdate.

[0014] In some embodiments, the molar ratio of the inorganic cerium salt to the molybdate is 1:4 to 1:10.

[0015] In some embodiments, drying the mixed solution comprises:

[0016] The mixed solution is placed in a controlled evaporation state, and a dark blue CMB solution is obtained after 2-4 days;

[0017] The CMB solution was freeze-dried and stored.

[0018] In some embodiments, drying the mixed solution comprises:

[0019] Take 5-6 ml of the mixed solution and transfer it to a 20 ml vial sealed with a metal cap having a rubber septum, pierce the rubber septum with a hollow needle with an inner diameter of 1.2 mm, and then place the vial in an oven and adjust the oven temperature to 95-100°C.

[0020] In some embodiments, drying the mixed solution comprises:

[0021] 5.4 ml of the mixed solution was transferred to a 20 ml vial sealed with a metal cap having a rubber septum, the rubber septum was pierced with a hollow needle with an inner diameter of 1.2 mm, and the vial was placed in an oven, and the oven temperature was adjusted to 100°C.

[0022] In some of the embodiments, including:

[0023] preparing a first mixed solution containing an inorganic cerium salt and a reducing agent;

[0024] adding an inorganic acid to the first mixed solution to obtain a second mixed solution;

[0025] adding molybdate to the second mixed solution to obtain a mixed solution;

[0026] The mixed solution is dried to obtain cerium-substituted molybdenum-based polyoxometalate.

[0027] In some of the embodiments, including

[0028] CeCl 3 7H 2 O and hydrazine hydrochloride are dissolved in deionized water to obtain a first mixed solution, wherein CeCl 3 7H 2 The ratio of the mass of O to the volume of deionized water is (3.6-4.4):1, and the ratio of the mass of hydrazine hydrochloride to the volume of deionized water is (2.7-3.3):5;

[0029] adding HCl to the first mixed solution to obtain a second mixed solution, wherein the concentration of the added HCl is 0.07-0.08 M;

[0030] Adding Na 2 MoO 4 ·2H 2 O to the second mixed solution to obtain a mixed solution, wherein Na 2 MoO 4 ·2H 2 The ratio of the mass of O to the volume of deionized water is (9-11):1;

[0031] The mixed solution is dried to obtain cerium-substituted molybdenum-based polyoxometalate.

[0032] In some of the embodiments, CeCl 3 7H 2 The ratio of the mass of O to the volume of deionized water is 4:1.

[0033] In some of the embodiments, the ratio of the mass of hydrazine hydrochloride to the volume of deionized water is 3:5.

[0034] In some of the embodiments, Na 2 MoO 4 ·2H 2 The ratio of the mass of O to the volume of deionized water is 10:1.

[0035] In some of the embodiments, including:

[0036] preparing a first solution containing an inorganic cerium salt;

[0037] preparing a second solution containing a reducing agent;

[0038] preparing a third solution containing molybdate;

[0039] mixing the first solution and the second solution to obtain a first mixed solution;

[0040] adding an inorganic acid to the first mixed solution to obtain a second mixed solution;

[0041] mixing the third solution with the second mixed solution to obtain a mixed solution;

[0042] The mixed solution is dried to obtain cerium-substituted molybdenum-based polyoxometalate.

[0043] In some of the embodiments, including:

[0044] CeCl 3 7H 2 O is dissolved in deionized water to obtain a first solution, wherein CeCl 3 7H 2 The ratio of the mass of O to the volume of deionized water is (18-22):1;

[0045] Dissolving hydrazine hydrochloride in deionized water to obtain a second solution, wherein the ratio of the mass of hydrazine hydrochloride to the volume of deionized water is (2.7-3.3):1;

[0046] Will Na 2 MoO 4 ·2H 2 O is dissolved in deionized water to obtain a third solution, wherein Na 2 MoO 4 ·2H 2 The ratio of the mass of O to the volume of deionized water is (45-55):1;

[0047] The first solution, the second solution and deionized water are mixed to obtain a first mixed solution, wherein the ratio of the volume of the deionized water, the volume of the first solution and the volume of the second solution is (1.8-2.2):(0.9-1.1):1;

[0048] adding HCl to the first mixed solution to obtain a second mixed solution, wherein the concentration of the added HCl is 0.09-0.11 M;

[0049] Mixing the third solution with the second mixed solution to obtain a mixed solution, wherein the ratio of the volume of the third solution to the volume of deionized water is (0.9-1.1):2;

[0050] The mixed solution is dried to obtain cerium-substituted molybdenum-based polyoxometalate.

[0051] In some of the embodiments, CeCl 3 7H 2 The ratio of the mass of O to the volume of deionized water is 20:1.

[0052] In some of the embodiments, the ratio of the mass of hydrazine hydrochloride to the volume of deionized water is 3:1.

[0053] In some of the embodiments, Na 2 MoO 4 ·2H 2 The ratio of the mass of O to the volume of deionized water is 50:1.

[0054] In some embodiments, the ratio of the volume of deionized water, the volume of the first solution, and the volume of the second solution is 2:1:1.

[0055] In some embodiments, the ratio of the volume of the third solution to the volume of deionized water is 1:2.

[0056] In a third aspect, there is provided a use of the material as described in the first aspect or the material prepared by the preparation method as described in the second aspect in the preparation of a drug for treating inflammatory skin diseases.

[0057] The present invention adopts the above technical solution, and has the following technical effects compared with the prior art:

[0058] The present invention provides a material, preparation method and application for treating inflammatory skin diseases. CMB has reliable biosafety and excellent reactive oxygen scavenging ability, can effectively inhibit the intracellular ROS level at a relatively low concentration, and thus inhibit the inflammatory activation of psoriasis or atopic dermatitis, thereby playing a role in treating psoriasis or atopic dermatitis. Compared with water-insoluble inorganic nanomaterials, CMB materials are soluble in water, are not easily retained in local tissues, are more easily metabolized by the body, and have better safety. BRIEF DESCRIPTION OF THE DRAWINGS

[0059] Figure 1 The morphology of the CMB solid prepared in Example 2 under a microscope;

[0060] Figure 2 TEM image of the CMB solution prepared in Example 2 (left) and element distribution diagram of Mo, Ce and O (right);

[0061] Figure 3 This is the XPS spectrum of the Mo element of the CMB prepared in Example 2;

[0062] Figure 4 The XPS spectrum of Ce element of CMB prepared in Example 2;

[0063] Figure 5 It is the DPPH test result in Example 5;

[0064] Figure 6 The ABTS test results in Example 5;

[0065] Figure 7 The ESR spectra of CMB in Example 5 were used to measure the effect of superoxide anion free radicals (O 2 · - )’s clearance capacity;

[0066] Figure 8 The results of the ESR spectrum measurement of the scavenging ability of CMB on intracellular hydroxyl free radicals (·OH) in Example 5;

[0067] Fig. 9 The results of using the DCFH-DA fluorescent probe to detect intracellular ROS levels in Example 5;

[0068] Fig.10 The results of the experiment on the inhibition of inflammatory response mediated by TNF-α in Example 6 are as follows;

[0069] Fig.11 This is a photograph showing the progress of IMQ-induced psoriasis-like skin inflammation in mice inhibited by subcutaneous injection of CMB in Example 7;

[0070] Fig.12 The results of H&E staining of skin sections of mice on the fifth day in Example 7;

[0071] Fig.13 The photographs show that subcutaneous injection of CMB inhibited the progression of MC903-induced atopic dermatitis in mice in Example 8;

[0072] Fig.14 This is the H&E staining result of the skin sections of mice on the sixth day in Example 8. DETAILED DESCRIPTION

[0073] In order to make the purpose, technical solutions and advantages of the present application clearer, the present application is described and illustrated below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and are not intended to limit the present application. Based on the embodiments provided in the present application, all other embodiments obtained by ordinary technicians in the field without making creative work are within the scope of protection of the present application.

[0074] Example 1

[0075] This embodiment relates to the materials, preparation methods and applications of the present invention for treating inflammatory skin diseases.

[0076] A material for treating inflammatory skin diseases, comprising cerium-substituted molybdenum-based polyoxometalate (CMB).

[0077] In the present invention, inflammatory skin diseases include psoriasis and atopic dermatitis.

[0078] For the above materials, the preparation method is as follows:

[0079] S1. Preparing a mixed solution containing an inorganic cerium salt and a molybdate, wherein the mixed solution is acidic;

[0080] S2. Dry the mixed solution to obtain cerium-substituted molybdenum-based polyoxometalate.

[0081] In step S1, the inorganic cerium salt includes cerium chloride, cerium nitrate, and cerium acetate.

[0082] In step S1, the molybdate includes sodium molybdate, potassium molybdate and ammonium molybdate.

[0083] In step S1, the molar ratio of the inorganic cerium salt to the molybdate is 1:4 to 1:10.

[0084] Furthermore, in step S1, it also includes:

[0085] A mixed solution containing an inorganic cerium salt, an inorganic acid and a molybdate is prepared.

[0086] In step S1, the inorganic acid includes but is not limited to HCl.

[0087] Furthermore, in step S1, it also includes:

[0088] A mixed solution containing an inorganic cerium salt, a reducing agent and a molybdate is prepared.

[0089] In step S1, the reducing agent includes but is not limited to hydrazine hydrochloride.

[0090] For step S1, it includes two implementation modes, which are as follows:

[0091] (I) First Implementation Method

[0092] S111. Preparing a first mixed solution containing an inorganic cerium salt and a reducing agent;

[0093] S112. Adding an inorganic acid to the first mixed solution to obtain a second mixed solution;

[0094] S113. Add molybdate to the second mixed solution to obtain a mixed solution.

[0095] (II) Second Implementation Method

[0096] S121. Preparing a first solution containing an inorganic cerium salt;

[0097] S122. Preparing a second solution containing a reducing agent;

[0098] S123. Preparing a third solution containing molybdate;

[0099] S124. Mixing the first solution with the second solution to obtain a first mixed solution;

[0100] S125. Adding an inorganic acid to the first mixed solution to obtain a second mixed solution;

[0101] S126. Mix the third solution with the second mixed solution to obtain a mixed solution.

[0102] In some of the embodiments, for the first implementation, including:

[0103] S111. CeCl 3 7H 2 O and hydrazine hydrochloride are dissolved in deionized water to obtain a first mixed solution, wherein CeCl 3 7H 2 The ratio of the mass of O to the volume of deionized water is (3.6-4.4):1, and the ratio of the mass of hydrazine hydrochloride to the volume of deionized water is (2.7-3.3):5;

[0104] S112. adding HCl to the first mixed solution to obtain a second mixed solution, wherein the concentration of HCl after addition is 0.07-0.08 M;

[0105] S113. Add Na 2 MoO 4 ·2H 2 O to the second mixed solution to obtain a mixed solution, wherein Na 2 MoO 4 ·2H 2 The ratio of the mass of O to the volume of deionized water is (9-11):1.

[0106] Preferably, in step S111, CeCl 3 7H 2 The ratio of the mass of O to the volume of deionized water is 4:1.

[0107] Preferably, in step S111, the ratio of the mass of hydrazine hydrochloride to the volume of deionized water is 3:5.

[0108] Preferably, in step S113, Na 2 MoO 4 ·2H2 The ratio of the mass of O to the volume of deionized water is 10:1.

[0109] In some of the embodiments, the second implementation includes:

[0110] S121. CeCl 3 7H 2 O is dissolved in deionized water to obtain a first solution, wherein CeCl 3 7H 2 The ratio of the mass of O to the volume of deionized water is (18-22):1;

[0111] S122. Dissolving hydrazine hydrochloride in deionized water to obtain a second solution, wherein the ratio of the mass of hydrazine hydrochloride to the volume of deionized water is (2.7-3.3):1;

[0112] S123. Na 2 MoO 4 ·2H 2 O is dissolved in deionized water to obtain a third solution, wherein Na 2 MoO 4 ·2H 2 The ratio of the mass of O to the volume of deionized water is (45-55):1;

[0113] S124. Mixing the first solution, the second solution and deionized water to obtain a first mixed solution, wherein the ratio of the volume of the deionized water, the volume of the first solution and the volume of the second solution is (1.8-2.2):(0.9-1.1):1;

[0114] S125. adding HCl to the first mixed solution to obtain a second mixed solution, wherein the concentration of HCl after addition is 0.09-0.11 M;

[0115] S126. Mix the third solution with the second mixed solution to obtain a mixed solution, wherein the ratio of the volume of the third solution to the volume of deionized water is (0.9-1.1):2.

[0116] Preferably, in step S121, CeCl 3 7H 2 The ratio of the mass of O to the volume of deionized water is 20:1.

[0117] Preferably, in step S122, the ratio of the mass of hydrazine hydrochloride to the volume of deionized water is 3:1.

[0118] Preferably, in step S123, Na 2 MoO 4 ·2H 2The ratio of the mass of O to the volume of deionized water is 50:1.

[0119] Preferably, in step S124, the ratio of the volume of deionized water, the volume of the first solution, and the volume of the second solution is 2:1:1.

[0120] Preferably, in step S126, the ratio of the volume of the third solution to the volume of the deionized water is 1:2. The deionized water is the deionized water in step S124.

[0121] Further, step S2 includes:

[0122] Step S21. The mixed solution is placed in a controlled evaporation state, and a dark blue CMB solution is obtained after 2-4 days;

[0123] Step S22: freeze-dry the CMB solution and store it.

[0124] Preferably, in step S21, a dark blue CMB solution is obtained after 3 days.

[0125] In some embodiments, step S21 includes:

[0126] Take 5-6 ml of the mixed solution and transfer it to a 20 ml vial sealed with a metal cap having a rubber septum. Pierce the rubber septum with a hollow needle with an inner diameter of 1.2 mm. Place the vial in an oven and adjust the oven temperature to 95-100°C.

[0127] Preferably, 5.4 ml of the mixed solution is transferred to a 20 ml vial sealed with a metal cap having a rubber septum, the rubber septum is pierced with a hollow needle with an inner diameter of 1.2 mm, and the vial is placed in an oven, and the oven temperature is adjusted to 100°C.

[0128] The embodiment of the present invention also provides a use of the cerium-substituted molybdenum-based polyoxometalate as described above in the preparation of a drug for treating inflammatory diseases.

[0129] Specifically, the inflammatory diseases include, but are not limited to: dermatomyositis, Grave's disease, multiple sclerosis, myasthenia gravis, systemic lupus erythematosus (SLE), sarcoidosis, Sjögren's syndrome, amyloidosis, Hashimoto's thyroiditis, vasculitis, rheumatoid arthritis, reactive arthritis, polymyositis, scleroderma, Addison's disease, vitiligo, pernicious anemia, glomerulonephritis, severe celiac disease, type 1 diabetes, psoriasis and pulmonary fibrosis, ulcerative colitis, Crohn's disease, atopic dermatitis.

[0130] Preferably, the inflammatory disease is psoriasis or atopic dermatitis.

[0131] In particular, the inflammatory disease is associated with elevated levels of inflammatory factors that stimulate cells to generate reactive oxygen species (ROS).

[0132] Specifically, the inflammatory factor is one or more of CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL8 and IL36G.

[0133] Specifically, the increased level of the inflammatory factor is caused by the stimulation of TNF-α.

[0134] Example 2 Preparation of Ce-Substituted Molybdenum-Based Polyoxometalate (CMB)

[0135] 20 mg CeCl 3 7H 2 O and 3 mg of hydrazine hydrochloride were dissolved in 5 ml of water, and then 0.4 ml of 1 M HCl was added with stirring;

[0136] While stirring, 50 mg Na 2 Mo 4 ·2H 2 O;

[0137] Continue stirring for 10-20 minutes to obtain a clear dark blue solution;

[0138] Then it was transferred to a 20 ml vial sealed with a metal cap having a rubber septum, and the rubber septum was pierced with a hollow needle with an inner diameter of 1.2 mm, so that the clear dark blue solution transferred to the bottle was placed in a state of controlled evaporation and heated at 100 °C in an oven to obtain a dark blue CMB solution after 3 days;

[0139] Store it freeze-dried.

[0140] Example 3 Preparation of cerium-substituted molybdenum-based polyoxometalate (CMB)

[0141] Weigh 20 mg of CeCl 3 7H 2 O, 3 mg hydrazine hydrochloride and 50 mg Na 2 MoO 4 ·2H 2 O, respectively dissolved in 1 ml of water to obtain 1 mL of each of the three solutions;

[0142] Take 2 mL of deionized water and add 1 mL of the aforementioned CeCl 3 solution and 1 mL of the aforementioned hydrazine hydrochloride solution;

[0143] While maintaining stirring, add 0.4 mL of 1 M HCl;

[0144] While maintaining stirring, add the aforementioned Na 2 MoO 4 1 mL of aqueous solution;

[0145] Continue stirring for 10-20 minutes to obtain a clear dark blue solution;

[0146] This was transferred to a 20-mL vial sealed with a metal cap with a rubber septum;

[0147] The rubber septum was pierced with a hollow needle with an inner diameter of 1.2 mm to put the sample in a state of controlled evaporation;

[0148] After heating at 100 °C in an oven for 3 days, a dark blue solution containing CMB was obtained;

[0149] It was freeze-dried and stored for later use.

[0150] Example 4 Characterization of Ce-substituted Molybdenum-based Polyoxometalates

[0151] The CMB solid stored after freeze-drying in Example 2 was observed under a light microscope. It can be observed through the microscope that the obtained CMB crystal has a typical rhombus morphology, which is consistent with the typical morphology of molybdenum-based POM crystals ( Figure 1 ).

[0152] Subsequently, CMB was dissolved in physiological saline to prepare CMB solution. TEM and element mapping were used to characterize the CMB solution. It can be seen that CMB spontaneously assembled into a round vesicle structure in the solution. Due to the drying step in the TEM sample preparation process, the vesicle structure often collapsed during observation, but the wrinkles on the circular surface left by the vesicle structure can still be observed. The circular edge observed by element mapping has high signals of Mo and Ce elements ( Figure 2 ).

[0153] Furthermore, XPS was used to characterize the elemental valence state of CMB solids, and it can be seen that the Mo element in CMB exists in a mixed valence state of +5 and +6 ( Figure 3 ), has potential free radical scavenging ability, while Ce is in a stable +3 valence state in CMB ( Figure 4 ).

[0154] Example 5 Evaluation of ROS scavenging activity of cerium-substituted molybdenum-based polyoxometalates

[0155] 1. The ROS scavenging activity of CMB was evaluated by DPPH and ABTS free radical scavenging assays using UV-visible spectroscopy:

[0156] DPPH test: pre-prepare 0.04 mg mL -1DPPH ethanol solution, then 2 mL of CMB solution with different concentrations was mixed with 2 mL of DPPH solution for 30 min, and then the absorbance at 519 nm was measured by UV-visible spectroscopy.

[0157] ABTS detection: 0.2 mL ABTS solution (7.4 mmol L -1 ) and 0.2 mL potassium persulfate solution (2.6 mmol L -1 ) and react in the dark for 24 h to generate ABTS. + ; then diluted 50 times with PBS buffer and reacted with CMB, 0.6 mL of CMB solution with different concentrations was mixed with 2.4 mL of ABTS· + After the solution was mixed and reacted for 10 min, the absorbance was measured at a wavelength of 734 nm.

[0158] Results: In DPPH( Figure 5 ) and ABTS test ( Figure 6 ) were observed to have a decreased absorbance at the characteristic peaks of free radicals (DPPH at 519 nm; ABTS at 724 nm), indicating that CMB has good free radical scavenging ability.

[0159] 2. The scavenging ability of CMBs on representative intracellular ROS, including superoxide anion radicals (O 2 · - )( Figure 7 ) and hydroxyl radicals (·OH)( Figure 8 ):

[0160] ESR spectrum detection: OH is generated by FeSO 4 and H 2 O 2 The Fenton reaction between 2 · - Produced by xanthine / xanthine oxidase reaction, TEMP and DMPO were selected as ·OH and O 2 · - of spin trapping agents.

[0161] Results: At 0, 5 and 10 minutes after the start of the reaction, O 2 · - After incubation with 100 μg / mL CMB for 10 min (reaction end point), the O 2 · - The characteristic peaks of ·OH and ·OH were sharply weakened to near the baseline level, indicating that CMB has a strong ROS elimination efficiency.

[0162] 3. CMB reduces intracellular ROS levels and inhibits the release of inflammatory factors:

[0163] The inhibitory effect of CMB on intracellular ROS level and inflammation was verified in keratinocytes (KC). CMB was co-incubated with KC cells and then treated with 400 μM H 2 O 2 The cells were treated for 1 h to establish the KC oxidative stress model, and then the DCFH-DA fluorescent probe was used to detect the ROS levels in KC cells with different treatments.

[0164] result( Fig. 9 ): In H 2 O 2 Under the induced oxidative stress conditions, CMB could significantly inhibit the increase of ROS levels in KC cells.

[0165] Example 6 Inhibition experiment of TNF-α mediated inflammatory response

[0166] HaCaT cells were evenly seeded in a 24-well plate. When the cells were attached and the density reached 60%, they were treated with different concentrations of CMB (30, 60, 90 μg / mL). Cell RNA was extracted after 1, 3, and 6 h of TNF-α (50 ng / mL) treatment, and the RNA levels of various inflammatory factors (CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL8 and IL36G) in the cells were detected by RT-qPCR.

[0167] result( Fig.10 ): It can be seen that the level of inflammatory factors in HaCaT cells treated with CMB was significantly inhibited after stimulation with TNF-α, and basically as the CMB dose increased, the RNA level of inflammatory factors gradually decreased, indicating that the excellent ROS scavenging ability of CMB can effectively inhibit TNF-α-mediated inflammatory response in vivo, and can effectively alleviate cell oxidative stress caused by external stimuli and inhibit the release of inflammatory factors.

[0168] Example 7 Ce-substituted molybdenum-based polyoxometalates inhibit IMQ-induced psoriasis-like skin inflammation in mice

[0169] First, an IMQ-induced psoriasis mouse model was constructed: male C57 mice aged 6-8 weeks were selected, and after depilation of the back, IMQ was applied topically at a dose of 30 mg per day, once every 24 hours, for a total of 5 days (day 0-day 4). On the 5th day, the mice were killed and samples were collected.

[0170] From the first day of application, the experimental group began to receive subcutaneous injection of CMB. The method was: CMB was injected subcutaneously at the site of IMQ application (diluted with saline, the concentration was 120μg / mL, 100μL was injected subcutaneously to each mouse, and the needle was inserted at multiple points during injection to try to make CMB evenly distributed subcutaneously); the control group was given sterile saline subcutaneous injection in the same way and frequency. The time interval between administration and modeling was at least 6h.

[0171] Results: From day 1 to day 5, subcutaneous injection of CMB effectively inhibited the progression of IMQ-induced psoriasis-like skin inflammation ( Fig.11 ). H&E staining of the skin sections of mice on day 5 also showed that the stratum corneum of the skin of the control mice was significantly thickened, while the thickness of the stratum corneum of the skin of the CMB-treated mice was significantly reduced ( Fig.12 ). Compared with the POM material, the dosage of CMB of the present invention is only 120 μg / mL, and each mouse is given a subcutaneous injection of 100 μL, which is much lower than the dosage mentioned in POM, indicating that the CMB of the present invention has a very strong free radical scavenging ability and a corresponding psoriasis treatment effect.

[0172] Example 8 Ce-substituted molybdenum-based polyoxometalates inhibit MC903-induced atopic dermatitis in mice

[0173] First, an MC903-induced atopic dermatitis mouse model was constructed: male C57 mice aged 6-8 weeks were selected, and after depilation of the back, IMQ was applied topically at 4 nmol per day, once every 24 hours, for a total of 5 days (Day 0-Day 4). On the 5th day, the mice were killed and samples were collected.

[0174] From the first day of application, the experimental group began to receive CMB subcutaneous injection treatment. The method was: CMB was subcutaneously injected at the MC903 application site (diluted with saline, the concentration was 120μg / mL, 100μL was subcutaneously injected to each mouse, and the needle was inserted at multiple points during the injection to try to make CMB evenly distributed under the skin); the control group was given sterile saline subcutaneously with the same administration method and frequency. The time interval between administration and modeling was at least 6h.

[0175] Results: From day 1 to day 6, subcutaneous injection of CMB effectively inhibited the progression of MC903-induced atopic dermatitis-like skin inflammation ( Fig.13 ). H&E staining of the skin sections of mice on day 6 also showed that the epidermis of the skin of the control mice was significantly thickened, while the thickness of the epidermis of the skin of the CMB-treated mice was significantly reduced ( Fig.14). Compared with the POM material, the dosage of the CMB of the present invention is only 120 μg / mL, and each mouse is given a subcutaneous injection of 100 μL, which is much lower than the dosage mentioned in POM, indicating that the CMB of the present invention has a very strong free radical scavenging ability and a corresponding atopic dermatitis treatment effect.

[0176] The above description is only a preferred embodiment of the present invention, and does not limit the implementation mode and protection scope of the present invention. For those skilled in the art, it should be aware that all solutions obtained by equivalent substitutions and obvious changes made using the description and illustrations of the present invention should be included in the protection scope of the present invention.

Claims

1. A material for treating inflammatory skin diseases, characterized in that: including cerium-substituted molybdenum-based polyoxometalates; Wherein, the inflammatory skin diseases include psoriasis and atopic dermatitis.

2. A preparation method for preparing the material for treating inflammatory skin diseases as claimed in claim 1, characterized in that: include: preparing a mixed solution containing an inorganic cerium salt and a molybdate, wherein the mixed solution is acidic; The mixed solution is dried to obtain cerium-substituted molybdenum-based polyoxometalate.

3. The preparation method according to claim 2, characterized in that: include: preparing a first mixed solution containing an inorganic cerium salt and a reducing agent; adding an inorganic acid to the first mixed solution to obtain a second mixed solution; adding molybdate to the second mixed solution to obtain a mixed solution; The mixed solution is dried to obtain cerium-substituted molybdenum-based polyoxometalate.

4. The preparation method according to claim 3, characterized in that: include Dissolving CeCl3·7H2O and hydrazine hydrochloride in deionized water to obtain a first mixed solution, wherein the ratio of the mass of CeCl3·7H2O to the volume of deionized water is (3.6-4.4):1, and the ratio of the mass of hydrazine hydrochloride to the volume of deionized water is (2.7-3.3):5; adding HCl to the first mixed solution to obtain a second mixed solution, wherein the concentration of the added HCl is 0.07-0.08 M; Adding Na2MoO4·2H2O to the second mixed solution to obtain a mixed solution, wherein the ratio of the mass of Na2MoO4·2H2O to the volume of deionized water is (9-11):1; The mixed solution is dried to obtain cerium-substituted molybdenum-based polyoxometalate.

5. The preparation method according to claim 2, characterized in that: include: preparing a first solution containing an inorganic cerium salt; preparing a second solution containing a reducing agent; preparing a third solution containing molybdate; mixing the first solution and the second solution to obtain a first mixed solution; adding an inorganic acid to the first mixed solution to obtain a second mixed solution; mixing the third solution with the second mixed solution to obtain a mixed solution; The mixed solution is dried to obtain cerium-substituted molybdenum-based polyoxometalate.

6. The preparation method according to claim 5, characterized in that: include: Dissolving CeCl3·7H2O in deionized water to obtain a first solution, wherein the ratio of the mass of CeCl3·7H2O to the volume of the deionized water is (18-22):1; Dissolving hydrazine hydrochloride in deionized water to obtain a second solution, wherein the ratio of the mass of hydrazine hydrochloride to the volume of deionized water is (2.7-3.3):1; Dissolving Na2MoO4·2H2O in deionized water to obtain a third solution, wherein the ratio of the mass of Na2MoO4·2H2O to the volume of the deionized water is (45-55):1; The first solution, the second solution and deionized water are mixed to obtain a first mixed solution, wherein the ratio of the volume of the deionized water, the volume of the first solution and the volume of the second solution is (1.8-2.2):(0.9-1.1):1; adding HCl to the first mixed solution to obtain a second mixed solution, wherein the concentration of the added HCl is 0.09-0.11 M; Mixing the third solution with the second mixed solution to obtain a mixed solution, wherein the ratio of the volume of the third solution to the volume of deionized water is (0.9-1.1):2; The mixed solution is dried to obtain cerium-substituted molybdenum-based polyoxometalate.

7. The preparation method according to any one of claims 2 to 6, characterized in that: Drying the mixed solution comprises: The mixed solution is placed in a controlled evaporation state, and a dark blue CMB solution is obtained after 2-4 days; The CMB solution was freeze-dried and stored.

8. The preparation method according to claim 7, characterized in that: Drying the mixed solution comprises: Take 5-6 ml of the mixed solution and transfer it to a 20 ml vial sealed with a metal cap having a rubber septum, pierce the rubber septum with a hollow needle with an inner diameter of 1.2 mm, and then place the vial in an oven and adjust the oven temperature to 95-100°C.

9. The preparation method according to any one of claims 2 to 6, characterized in that: The inorganic cerium salt includes cerium chloride, cerium nitrate, cerium acetate; and / or The molybdate includes sodium molybdate, potassium molybdate, ammonium molybdate; and / or The molar ratio of the inorganic cerium salt to the molybdate is 1:4 to 1:

10.

10. Use of the material according to claim 1 or the material prepared by the preparation method according to any one of claims 2 to 9 in preparing a drug for treating inflammatory skin diseases.

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