Traditional Chinese medicine nanoparticles for treating psoriasis and application thereof

Abstract

The invention relates to the field of biological medicine, in particular to traditional Chinese medicine nano-particles for treating psoriasis and application of the traditional Chinese medicine nano-particles. The traditional Chinese medicine nanoparticles are prepared according to the following steps: mixing an aqueous solution of cortex dictamni with a buffer solution, adding Pluronic F-127 to wrap the mixture, and then coupling with ferric chloride hexahydrate to obtain the traditional Chinese medicine nanoparticles. The water solubility of the traditional Chinese medicine nano-particles is remarkably improved, the traditional Chinese medicine nano-particles can be well absorbed by keratinocytes and mice, and a good psoriasis treatment effect is achieved.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a kind of traditional Chinese medicine nanoparticles for treating psoriasis and application thereof. Background Art

[0002] Psoriasis is a skin disease associated with immune activation. Its pathological mechanism involves a complex interaction between the innate and adaptive immune systems. Histological features include excessive proliferation and abnormal differentiation of keratinocytes (KC), dermal inflammatory infiltration, and angiogenesis. Psoriasis has a high incidence rate and is increasing year by year. Its recurrence rate can be as high as 98.4%, and it is often accompanied by chronic diseases, disfigurement, disability, and related complications, which has a serious impact and burden on the physical and mental health of patients.

[0003] At present, the treatment methods of psoriasis mainly include topical medications, phototherapy, systemic treatment, traditional Chinese medicine treatment and other auxiliary treatments. Topical medications include glucocorticoids, vitamin D3 derivatives, retinoic acid drugs, calcineurin inhibitors, etc., which are suitable for mild to moderate patients and can relieve symptoms, but biological agents still have problems such as poor effect in some patients, increased risk of infection, formation of immune tolerance, long-term tumor risk, and monoclonal antibody-related psoriasis outbreaks and recurrences. Phototherapy includes ultraviolet B phototherapy, PUVA therapy and excimer laser, which improve skin lesions through light irradiation, but the dose and course of treatment need to be controlled. Therefore, in response to the medical needs of psoriasis, the development of targeted drugs to improve patients' health status and quality of life is the key to psoriasis research.

[0004] my country has used traditional Chinese medicine to intervene in the development of psoriasis for thousands of years, and during this period, it has accumulated rich experience and formed a unique theory of TCM treatment of psoriasis. A variety of traditional Chinese herbal medicines have been reported to effectively alleviate the occurrence and development of psoriasis, including inhibiting the rapid proliferation of keratinocytes and reducing the expression of inflammatory factors. However, the unclear targets of traditional Chinese medicine and the differences in ingredients between different batches are the key obstacles to the entry of traditional Chinese medicine into the world's medical system. Dictamni is a traditional Chinese medicine in my country. Related records have been made in "Shennong's Herbal Classic". In the Chinese Pharmacopoeia, there are dozens of medicines containing Dictamni for the treatment of skin diseases. Dictamni can be taken orally, applied externally or locally by injection. It has been proven to have the effects of clearing away heat and detoxifying, removing dampness and relieving itching in skin diseases. It is widely used to treat tinea pedis, urticaria, eczema, psoriasis, flat warts, skin cancer, etc. See. Figure 2 A. However, the poor water solubility of Dictamni affects its development and application. Summary of the invention

[0005] Based on the above technical problems, the present invention coordinates the Dictamni cortex drug particles with iron ions and forms a Dictamni cortex loaded nanoparticle carrier under the action of Pluronic F-127, which has significantly improved water solubility, can be well absorbed by keratinocytes and mice, and exerts a good psoriasis treatment effect.

[0006] The specific technical solutions provided by the present invention are as follows: In a first aspect, the present invention provides a Chinese medicine nanoparticle for treating psoriasis, which is prepared according to the following steps: The aqueous solution of Dictamni cortex is mixed with a buffer solution, Pluronic F-127 is added to encapsulate the mixture, and then the mixture is coupled with ferric chloride hexahydrate to obtain the traditional Chinese medicine nanoparticles.

[0007] The buffer solution is used to adjust the pH value to control the reaction rate and particle stability.

[0008] As a preferred embodiment of the present invention, the aqueous solution of Dictamni cortex is prepared according to the following steps: Dictamni cortex particles are uniformly dispersed in water.

[0009] As a preferred embodiment of the present invention, the buffer is Tris buffer or 4-hydroxyethylpiperazineethanesulfonic acid or PBS solution or borate solution.

[0010] As a preferred embodiment of the present invention, after mixing the Tris buffer with the aqueous solution of Dictamni cortex, gradually adding the ferric chloride hexahydrate aqueous solution and the Pluronic F-127 solution, mixing and dispersing, to obtain the traditional Chinese medicine nanoparticles; The concentration of Tris buffer is 10mM~100mM, the pH is 8.0~9.0, the concentration of the aqueous solution of Dictamni cortex is 5mg / mL~20mg / mL, the concentration of the aqueous solution of ferric chloride hexahydrate is 5mg / mL~50mg / mL, the mass concentration of Pluronic F-127 is 0.1%~5%, and the volume ratio of Tris buffer, aqueous solution of Dictamni cortex, aqueous solution of ferric chloride hexahydrate and Pluronic F-127 solution is 25:1:1:4.

[0011] As a preferred embodiment of the present invention, the dispersion is carried out by ultrasonic treatment at 30% power for 2 to 3 times. After ultrasonic treatment, the particles can be evenly distributed to prevent agglomeration and precipitation of particles.

[0012] As a preferred embodiment of the present invention, the particle size of the traditional Chinese medicine nanoparticles is 34.94-35.58 nm.

[0013] The second aspect of the present invention provides a use of the traditional Chinese medicine nanoparticles in preparing a drug for treating psoriasis.

[0014] The third aspect of the present invention provides a drug for treating psoriasis, wherein the drug contains the traditional Chinese medicine nanoparticles as the only active ingredient.

[0015] As a preferred embodiment of the present invention, the drug is prepared by compounding the traditional Chinese medicine nanoparticles with pharmaceutically acceptable excipients.

[0016] As a preferred embodiment of the present invention, the drug is an oral preparation, an external preparation or an injection preparation.

[0017] Compared with the prior art, the beneficial effects of the present invention are: (1) Pluronic F-127 is a nonionic surfactant polyol that can promote the dissolution of other substances in physiological media. In the present invention, it is used to wrap the water-soluble drug particles of Dictamni cortex. Dictamni cortex has poor water solubility and will precipitate after being left at room temperature. The addition of Pluronic F-127 greatly enhances the water solubility and bioavailability, and can continue to remain stable in physiological environments such as PBS and culture medium. At the same time, ferric chloride hexahydrate (hexacoordinated) is used to couple Pluronic F-127 wrapped with Dictamni cortex. The iron ion content in the skin and blood of psoriasis patients is reduced. By coupling iron ions, the iron content in the skin of the psoriasis mouse model can be significantly increased. In addition, Cl - The internal environment of the body objectively exists, so the use of ferric chloride hexahydrate will not introduce other elements and is highly safe.

[0018] (2) The Chinese medicine nanoparticles for treating psoriasis provided by the present invention have the effects of clearing away heat and detoxification, anti-inflammatory, and antipruritic. The present invention uses imiquimod-induced psoriasis mice and M5 (a mixture of multiple cytokines)-induced keratinocyte psoriasis cell models as disease models to verify its therapeutic effect on psoriasis and related molecular mechanisms. Based on the functions of this Chinese medicine nanotechnology, its application can be extended to clinical practice.

[0019] (3) The Chinese medicine nanoparticles for treating psoriasis provided by the present invention have good safety and no toxic side effects on the body. The coordinated iron ions can effectively improve the skin problems caused by iron ion deficiency in psoriasis skin.

[0020] (4) The present invention uses experiments combined with bioinformatics analysis to clarify the mechanism of action of Dictamni cortex in the treatment of psoriasis. It is verified that after Dictamni cortex acts on keratinocytes or mice, it will cause a decrease in HSP90AB1 and its binding protein CDC37. HSP90AB1 is a subunit of HSP90. HSP90 is widely highly expressed in inflammatory diseases and cancers, and its client proteins are as high as 100. The reduction of HSP90AB1 reduces the phosphorylation of client proteins Akt and stat3, effectively alleviating the occurrence of inflammation. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 : Characterization of loaded Dictamni cortex nanoparticles (NB). A. Changes in dissolution of Dictamni cortex in water within 72 h; B. Schematic diagram of NB synthesis; C. TEM image of NB particles; D. Lorenz TEM image of NB particles (scale = 100 nm, located on the left side of the first horizontal row) and energy dispersive spectrometer (EDS); E~F. Water and particle size (E) and Zeta potential (F) of NB; G~H. Changes in particle size of NB particles in PBS solution (G) and culture medium (H) within 7 days;

[0022] Figure 2 : Characterization of loaded Dictamni cortex nanoparticles (NB). A. Fourier transform infrared (FTIR) spectrum of NB particles; B. UV spectrum of NB particles; C~D. X-ray photoelectron spectroscopy (XPS) of NB particles, including Fe2p spectrum (C) and O1s spectrum (D); E. O1s spectrum of Dictamni cortex; F. Fe2p spectrum of FeCl3;

[0023] Figure 3 :Characterization of Dictamni cortex nanoparticles (NB). A~B. Images of spontaneous fluorescence of NB particles under ultraviolet light excitation (A) and fluorescence spectrum (B); C~D. Cumulative release of Dictamni cortex (C) and Fe by NB particles at different time points under different pH environments (pH=5.5, 6.8, 7.4). 3+ Quantification (D). Data represent mean ± SEM from at least three independent experiments. ***p < 0.001, **p < 0.01, *p < 0.05.

[0024] Figure 4: Dictamni cortex inhibits M5-induced HEKa cell proliferation and inflammatory response. A. The historical origin of Dictamni cortex and the current status of Chinese patent applications. B. Effects of different concentrations of Dictamni peel on HEKa cell viability after 24 h treatment; C. 20 mg / L Dictamni peel can inhibit M5-induced cell proliferation; D. Core Gene Ontology (GO) biological processes of luteolin, wogonin and quercetin, the main components of Dictamni peel; E. Molecular docking analysis of luteolin, wogonin and quercetin, the main components of Dictamni peel, with IL-6 / TNF; F. Venn diagram analysis of common molecular targets of luteolin, wogonin and quercetin; G. Molecular docking model of luteolin, wogonin and quercetin with IL-6 and TNF; H. Western blotting detection of p-NF-κB / NF-κB and p-STAT3 / STAT3 in M5 and Dictamni peel treatment groups; I. Immunofluorescence localization of p-STAT3 after Dictamni peel and M5 co-treatment for 24 h (scale bar = 10 μm); J. Relative expression levels of inflammatory factors (IL-1α, IL-1β, IL-17A, IL-17F, TNFα) mRNA; K. Relative expression levels of antioxidant system (HO-1, NQO-1, SOD1 / 2) mRNA; Data are expressed as mean ± standard error (n=3 or 6 groups of independent biological samples), *p<0.05, **p<0.01, ***p<0.001, ns, no statistical difference, two-tailed t-test / two-way analysis of variance combined with Bonferroni correction.

[0025] Figure 5 NB inhibits abnormal cell proliferation and inflammatory response. A. Schematic diagram of the experimental process of HEKa cells treated with M5 and NB; B. Quantitative analysis of cell proliferation in different treatment groups at 24 / 48 / 72 h by CCK8 method; C. Immunoblotting detection of p-NF-κB / NF-κB and p-STAT3 / STAT3 in M5 / NB treatment group; D. DHE fluorescence staining to trace intracellular ROS level, scale bar = 50 μm; E. DCFH-DA probe method to detect ROS generation; F. Relative expression levels of inflammatory-related cytokines IL-1α, IL-1β, IL-6, and TNFα mRNA; Data are expressed as mean ± standard error (n = 3 or 6 independent biological samples), *p < 0.05, **p < 0.01, ***p < 0.001, ns, no statistical difference, two-tailed t test / two-way ANOVA combined with Bonferroni correction.

[0026] Figure 6: Subcutaneous administration of NB alleviates psoriatic dermatitis in mice. A. Schematic diagram of the experimental process; B. Typical images of the phenotype of each group of mice on day 0 / 3 / 5 of treatment; C. Fluorescence microscopic imaging of skin tissue in the NB-treated group; D. Psoriasis area and severity index score; E. Curve of mouse weight changes during the experiment; F. H&E stained pathological sections of the back skin of each group of mice; G. Quantitative analysis of epidermal thickness based on H&E results; H. Morphological observation of spleen tissue in each group; I. Relative changes in spleen-body ratio (spleen weight / body weight) (standardized to the control group); J. Colorimetric detection of total iron content (OD 593nm )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。 )。

[0027] Figure 7: NB-triggered changes in nascent proteomics identified HSP90AB1 as a target; A. Schematic diagram of Aha labeling of nascent proteins (left) and mass spectrometry detection (right); B. Major biological processes of the enriched nascent proteome; C~D. Heatmaps© and volcano plots (D) of nascent proteins significantly changed after NB treatment compared with the Ctrl group; E. Molecular docking simulations of HSP90AB1 with luteolin, wogonin and quercetin, the upper row is the structural schematics of luteolin, wogonin and quercetin; the lower row is the molecular docking simulation, and the right side of each small figure is the enlarged image of the corresponding red frame on the left; F. Immunoblotting results of HSP90AB1 protein in IMQ-induced mouse skin (vs. control group); G. HSP90AB1 expression level results in the GEO database (GSE13355, GSE114286, GSE53431); H. Immunofluorescence staining of HSP90AB1 in skin tissues of healthy donors and psoriasis patients. The right panel is an enlarged view of the white frame on the left, and the scale bar = 100 μm. Data are presented as mean ± SEM (n = 3 biologically independent cell samples or 6 mice per group). The threshold for mass spectrometry analysis was set as |log2 fold change| ≥ 1 and p value < 0.05. *p < 0.05, **p < 0.01, ***p < 0.001, ns indicates no significance. Statistical analysis was performed using a two-tailed Student t test or a two-way ANOVA combined with a Bonferroni post hoc test.

[0028] Figure 8 : HSP90 inhibitor AUY922 improves IMQ-induced psoriasis in mice. A. Representative skin lesion images of four groups of mice on day 0, day 3, and day 5; B. Changes in mouse body weight and (C) Psoriasis Area and Severity Index (PASI) scores (divided into Ctrl group, IMQ group, IMQ+NB group, and IMQ+AUY922 group); D. Gross images of spleen and (E) spleen weight / body weight ratio (normalized to control group); F. H&E staining (first row), HSP90AB1 immunohistochemical staining (second row), and immunofluorescence analysis of the expression levels of HSP90AB1 (third row) and PCNA (fourth row) in skin tissues of four groups of mice; scale bar = 100 μm, data are expressed as mean ± SEM (n = 6 mice per group). *p < 0.05, **p < 0.01, ***p < 0.001, ns indicates no significance, and two-tailed t-test was used for statistical analysis. DETAILED DESCRIPTION

[0029] The present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. The embodiments are only used to explain the purpose, technical solutions and advantages of the present invention, but are not intended to limit the scope of protection of the present invention.

[0030] Psoriasis is a skin disease associated with immune activation. It has a high incidence rate and is increasing year by year. The recurrence rate can be as high as 98.4%, and it is often accompanied by chronic diseases, disfigurement, disability and related complications, which has a serious impact and burden on the physical and mental health of patients.

[0031] At present, the treatment methods of psoriasis mainly include topical medications, phototherapy, systemic treatment, traditional Chinese medicine treatment and other auxiliary treatments. Topical medications include glucocorticoids, vitamin D3 derivatives, retinoic acid drugs, calcineurin inhibitors, etc., which are suitable for mild to moderate patients and can relieve symptoms, but biological agents still have problems such as poor effect in some patients, increased risk of infection, formation of immune tolerance, long-term tumor risk, and monoclonal antibody-related psoriasis outbreaks and recurrences. Phototherapy includes ultraviolet B phototherapy, PUVA therapy and excimer laser, which improve skin lesions through light irradiation, but the dose and course of treatment need to be controlled. Therefore, in response to the medical needs of psoriasis, the development of targeted drugs to improve patients' health status and quality of life is the key to psoriasis research.

[0032] Dictamni is a traditional Chinese medicine in my country. It has been proven to have the effects of clearing away heat and detoxifying, removing dampness and relieving itching in skin diseases. It is widely used to treat tinea pedis, urticaria, eczema, psoriasis, flat warts, skin cancer, etc. However, the poor water solubility of Dictamni affects its development and application.

[0033] Based on this, the present invention provides a Chinese medicine nanoparticle for treating psoriasis, which is prepared according to the following steps: The aqueous solution of Dictamni cortex is mixed with a buffer solution, Pluronic F-127 is added to encapsulate the mixture, and then the mixture is coupled with ferric chloride hexahydrate to obtain the traditional Chinese medicine nanoparticles.

[0034] Pluronic F-127 is a nonionic surfactant polyol that can promote the dissolution of other substances in physiological media. In the present invention, it is used to wrap the water-soluble drug particles of Dictamni cortex. Dictamni cortex has poor water solubility and will precipitate after being placed at room temperature. After adding Pluronic F-127, the water solubility and bioavailability are greatly enhanced, and it can continue to remain stable under physiological environments such as PBS and culture medium. At the same time, ferric chloride hexahydrate (hexacoordination) is used to couple Pluronic F-127 wrapped with Dictamni cortex. The iron ion content in the skin and blood of psoriasis patients is reduced. By coupling iron ions, the iron content in the skin of the psoriasis mouse model can be significantly increased.

[0035] Example 1 Nanoparticles loaded with Dictamni cortex are synthesized using Dictamni cortex (traditional Chinese medicine), Pluronic F-127, ferric chloride hexahydrate, and Tris buffer system as raw materials, including the following preparation steps (taking 5 ml system as an example), such as Figure 1 As shown in B: (1) Place 5 ml of 10 mM Tris buffer on a magnetic stirrer and add 200 μL of 10 mg / mL Dictamni aqueous solution while rotating; (2) Add 200 μL of 10 mg / mL ferric chloride hexahydrate dropwise; (3) Add 200 μL of 2% Pluronic F-127 dropwise; (4) 30% power ultrasound 3 times; (3) Continue stirring for 1 to 2 days.

[0036] The Dictamni cortex aqueous solution was prepared according to the following steps: 50 mg of Dictamni cortex particles were weighed using an analytical balance, 5 mL of ddH2O was added, and the mixture was fully shaken to be uniformly dispersed in water. Figure 1 As shown in A.

[0037] After that, dialyze with MWCO3500 ultrafiltration membrane or ultrafiltration tube to make the remaining system about 1mL, and store it after ultrasound. When used, dissolve it in 0.9% NaCl aqueous solution and filter it with 0.45μm pore size filter membrane. After filtration, it can be directly injected locally. The obtained nanoparticles loaded with Dictamni are marked as NB particles.

[0038] Example 2 Nanoparticles loaded with Dictamni cortex are prepared according to the following steps: (1) Place 5 ml of 10 mM Tris buffer on a magnetic stirrer and add 200 μL of 5 mg / mL Dictamni aqueous solution while rotating; (2) Add 200 μL of 50 mg / mL ferric chloride hexahydrate dropwise; (3) Add 200 μL of 0.1% Pluronic F-127 dropwise; (4) 30% power ultrasound 3 times; (5) Continue stirring for 1 to 2 days.

[0039] The Dictamni Cortex aqueous solution was prepared according to the following steps: 50 mg of Dictamni Cortex particles were weighed using an analytical balance, 5 mL of ddH2O was added, and the mixture was fully shaken to be uniformly dispersed in the water.

[0040] After that, dialyze with MWCO3500 ultrafiltration membrane or ultrafiltration tube to make the remaining system about 1mL, and store it after ultrasound. When used, dissolve it in 0.9% NaCl aqueous solution and filter it with 0.45μm pore size filter membrane. After filtration, it can be directly injected locally. The obtained nanoparticles loaded with Dictamni are marked as NB particles.

[0041] Example 3 Nanoparticles loaded with Dictamni cortex are prepared according to the following steps: (1) Place 5 ml of 20 mM Tris buffer on a magnetic stirrer and add 200 μL of 10 mg / mL Dictamni aqueous solution while rotating; (2) Add 200 μL of 50 mg / mL ferric chloride hexahydrate dropwise; (3) Add 200 μL of 5% Pluronic F-127 dropwise; (4) 30% power ultrasound 3 times; (5) Continue stirring for 1 to 2 days.

[0042] The Dictamni Cortex aqueous solution was prepared according to the following steps: 50 mg of Dictamni Cortex particles were weighed using an analytical balance, 5 mL of ddH2O was added, and the mixture was fully shaken to be uniformly dispersed in the water.

[0043] After that, dialyze with MWCO3500 ultrafiltration membrane or ultrafiltration tube to make the remaining system about 1mL, and store it after ultrasound. When used, dissolve it in 0.9% NaCl aqueous solution and filter it with 0.45μm pore size filter membrane. After filtration, it can be directly injected locally. The obtained nanoparticles loaded with Dictamni are marked as NB particles.

[0044] Example 4 Nanoparticles loaded with Dictamni cortex are prepared according to the following steps: (1) Place 5 ml of 50 mM Tris buffer on a magnetic stirrer and add 200 μL of 10 mg / mL Dictamni aqueous solution while rotating; (2) Add 200 μL of 25 mg / mL ferric chloride hexahydrate dropwise; (3) Add 200 μL of 2% Pluronic F-127 dropwise; (4) 30% power ultrasound 3 times; (5) Continue stirring for 1 to 2 days.

[0045] The Dictamni Cortex aqueous solution was prepared according to the following steps: 50 mg of Dictamni Cortex particles were weighed using an analytical balance, 5 mL of ddH2O was added, and the mixture was fully shaken to be uniformly dispersed in the water.

[0046] After that, dialyze with MWCO3500 ultrafiltration membrane or ultrafiltration tube to make the remaining system about 1mL, and store it after ultrasound. When used, dissolve it in 0.9% NaCl aqueous solution and filter it with 0.45μm pore size filter membrane. After filtration, it can be directly injected locally. The obtained nanoparticles loaded with Dictamni are marked as NB particles.

[0047] Since the structures, properties and effects of the NB particles prepared in Examples 1 to 4 are basically similar, the present invention will be described below using only the NB particles prepared in Example 1 as an example.

[0048] TEM images ( Figure 1 The results of C) and dynamic light scattering (DLS) show that the NB particles are spherical or nearly spherical, with a relatively uniform distribution and no obvious agglomeration. The NB particles are bean-shaped with a hydrodynamic diameter of 35.26 ± 0.32 nm (consistent with the "NB (35.26 ± 0.32 nm)" marked in E). Lorenz TEM image ( Figure 1 Middle D) further shows the lattice fringes of the particles, indicating that they are nanoparticles with good crystallinity. EDS spectrum ( Figure 1 D) shows that the main elements are Fe and O, and the Fe 2+ / Fe 3+ The mixed valence state is consistent. No impurity elements such as C and Si were detected, indicating that the sample is of high purity.

[0049] The hydrated particle size and Zeta potential of NB and Dictamni were further analyzed. Figure 1 DLS further proves that BXP and Fe 3+ The Zeta potentials of NB and BXP were -19.5 mV and 10.3 mV, respectively, while the NB showed a narrow distribution of -11.8 mV after the positive ions were coordinated with BXP, indicating that NB was well dispersed in the solution and no significant agglomeration occurred, which was consistent with the TEM observation results. In addition, the Lorenz TEM and energy dispersive spectroscopy (EDS) results showed that carbon (C), oxygen (O) and iron (Fe) were uniformly distributed in NB.

[0050] The particle size of NB in ​​PBS and DMEM culture medium was tested during cell culture, and the results showed that NB could remain stable for 7 consecutive days ( Figure 1 (middle G~H).

[0051] Compared with Dictamni itself, the Fourier transform infrared (FTIR) spectrum of NB confirmed the presence of BXP in NB, such as 1103 cm -1 and 1119 cm -1 The peak at 400 nm confirmed the presence of BXP in NB. Figure 2 For example, 2883 cm -1 The peak at indicates that the proton is 3+ The OH stretching vibration is significantly weakened, while the 1103cm -1 The CO peak at 3+ During coordination, the electron cloud of the oxygen atom moves toward the metal ion, resulting in a decrease in the polarity of the CO bond, a shift of the peak position to a lower wave number, and a decrease in the peak intensity. ~580 cm -1 :Fe-O vibration peak, corresponding to the characteristic absorption of iron oxide. ~1630 cm -1 and 3400 cm -1 : Corresponding to the OH bending vibration and stretching vibration of adsorbed water, respectively, indicating the presence of hydroxyl groups (-OH) or adsorbed water molecules on the particle surface. ~1400 cm -1 : Possible CO stretching vibration (such as carbonate or carboxylate groups), but the peak intensity is weak, which may be related to trace organic contamination or precursor residues on the surface. No Oh-CO peak related to 332.5 eV was observed (combined with XPS data), indicating that the surface modification is mainly hydroxyl groups without significant organic ligand coverage.

[0052] Ultraviolet-visible (UV-Vis) absorption spectroscopy reconfirmed the presence of BXP in NB ( Figure 2 In addition, the Dictamni cortex itself has strong UV-visible absorption, which will cause signal interference in the biological effect experiment. The UV-visible absorption is reduced through NB assembly and can be kept stable.

[0053] In addition, X-ray photoelectron spectroscopy (XPS) was used to analyze the energy level changes of iron in NB and Dictamni cortex. Fe2p is the most common Auger spectrum peak of iron. In addition to the main peak, there is also a half peak (satellite). The high-resolution Fe 2p spectrum shows that Fe 2p 3 / 2 and Fe 2p 1 / 2 are 710.19 eV and 724.65 eV in NB, respectively, which indicates that Fe is widely present in NB. 3+ ( Figure 2 C). The high-resolution Fe 2p spectrum of FeCl3 shows that Fe 2p 3 / 2 and Fe 2p 1 / 2 are at 711.82 eV and 725.25 eV ( Figure 2The high-resolution O 1S spectrum of NB shows characteristic peaks at 533.52 eV and 532.13 eV ( Figure 2 (D), the high-resolution O 1S spectrum of BXP shows characteristic peaks at 530.98 eV and 532.73 eV ( Figure 2 F). This indicates that Fe + The electron pair of O atom is shared with BXP, and O atom is also coordinated to Fe 3+ The electron donation effect of ions results in a higher binding energy for O 1s and a lower binding energy for Fe 2p (the NB spectrum shows a higher binding energy for Fe 3+ , indicating the existence of a strong ligand field effect (hexacoordinated ferric chloride hexahydrate).

[0054] Dictamni and NB can produce autofluorescence after ultraviolet excitation, which may be caused by some compounds in Dictamni. The fluorescence spectrum detected the maximum excitation wavelength (Ex) of Dictamni and NB at 405 nm and the maximum emission wavelength (Em) at 468 nm, see Figure 3 Middle A~B.

[0055] The release of Dictamni and Fe by NB at different pH environments of pH = 5.5, 6.8, and 7.4 was further tested. 3+ In the psoriasis lesion area, lactic acid accumulates to maintain the inflammatory acidic environment. The results showed that ( Figure 3 (C-D) At pH 5.5, NB releases Dictamni and Fe 3+ Significant enhancement, with more than 80% of the drug released in the same amount of time, was much higher than in pH 6.8 and pH 7.4 environments, confirming the pH responsiveness of NB, indicating that it remains stable in a neutral pH environment and has targeting effects and anti-inflammatory properties.

[0056] Experimental Example 1 Bioinformatics analysis (1) Molecular docking The molecular structures of woflavin, luteolin, and quercetin, the core components of D. difficile cortex, were obtained from the PubChem database (https: / / pubchem.ncbi.nlm.nih.gov / ) and energy minimized using Chem3D software.

[0057] The 3D X-ray crystal structures of the key targets HSP90AB1, TNFα, and IL-6 were obtained from the PDB database (http: / / www.rcsb.org).

[0058] Both component and protein files were converted to PDBQT format. Molecular docking was performed using AutodockVina v1.2.2 software to obtain binding energy results. Finally, PYMOL software was used to visualize the docking results.

[0059] (2) Cluster analysis The online database Metascape was used to perform cluster analysis on the target proteins of woflavin, luteolin, and quercetin, and the GO results and pathway enrichment results were output.

[0060] Through the analysis of various component targets of Dictamni cortex, the most significant changes in several GO biological processes were summarized ( Figure 4 D) and key pathways of change ( Figure 4 Middle E). The common molecular targets of the three key ingredients, luteolin, wogonin and quercetin, were further analyzed using a Venn diagram ( Figure 4 F). Through molecular docking, it was found that luteolin, wogonin and quercetin can dock with IL-6 and TNF ( Figure 4 Middle G).

[0061] Experimental Example 2 Cell experiments verify the therapeutic effect of nanoparticles loaded with Dictamni on psoriasis 1. Cell culture and treatment HEKa cells were cultured in DMEM complete medium containing 10% fetal bovine serum (10% by volume) in an incubator at 37°C and 5% CO2. 10 ng / ml M5 was used to simulate psoriasis cell inflammation, which included IL-1α, IL-17A, IL-22, oncostatin M, and tumor necrosis factor-α.

[0062] 2. CCK-8 cell viability assay The experimental process is shown in Figure 5 Middle A.

[0063] (1) Cell inoculation: HEKa cells in the logarithmic growth phase were adjusted to a density of 5 × 10³ cells / well and inoculated into a 96-well plate with a volume of 100 μL per well. The cells were pre-cultured in a 37°C, 5% CO2 incubator for 12 h.

[0064] (2) Drug treatment: The original culture medium was discarded and fresh culture medium containing different concentrations of Dictamni or NB nanoparticles was added. Six replicate wells were set up in each group and the cells were treated for 24, 48, and 72 h, respectively.

[0065] (3) Color development reaction: Add 10 μL of CCK-8 solution to each well and incubate for 2 h in the dark.

[0066] (4) Absorbance detection: The absorbance at 450 nm was measured by an ELISA instrument with a reference wavelength of 650 nm, and the background value of the blank well was subtracted.

[0067] (5) Data analysis: Cell viability (%) = (OD value of experimental group - OD value of blank) / (OD value of control group - OD value of blank) × 100%. The results are expressed as mean ± SEM.

[0068] 3. Total RNA extraction, reverse transcription and qRT-PCR from cell tissues Total RNA of cell tissues was extracted and reverse transcribed, and the reverse transcribed products were subjected to PCR to detect the expression of specific genes. The reverse transcription system is shown in Table 1, and the PCR reaction system is shown in Table 2.

[0069] Table 1 Reverse transcription system Reverse transcription conditions were: 25°C, 10 min; 42°C, 30 min; 85°C, 5 min; Table 2 PCR system The PCR reaction conditions are as follows: ⑨ Obtain the melting curve of the corresponding gene through the process of ⑥~⑧ and normalize the gene expression value to GAPDH / Gapdh, and use 2 −∆∆Ct Method for analysis.

[0070] 4. Total cell protein extraction and Western blot detection IP lysis was used to extract total cell protein, followed by BCA quantification and protein concentration adjustment. After that, the protein was detected by Western blot, using primary antibodies and corresponding secondary antibodies against p-NF-κB / NF-κB and p-STAT3 / STAT3. Finally, ECL luminescence was used to detect the grayscale value of protein bands, and the results were quantitatively analyzed.

[0071] 5. Cell Immunofluorescence Staining (1) Inoculate the cells into a well plate with a glass slide. When the cells grow to an appropriate density, wash the glass slide with PBS three times, 3 minutes each time.

[0072] (2) Fix with 4% paraformaldehyde and wash with PBS three times, 3 minutes each time.

[0073] (3) 0.5% Triton X-100, permeabilization for 20 minutes.

[0074] (4) Wash with PBS three times, 3 minutes each time, aspirate the PBS, add goat serum, and block for 30 minutes.

[0075] (5) Aspirate the blocking solution, add primary antibody without washing, and incubate at 4°C overnight.

[0076] (6) Wash with PBST three times for 3 minutes each time, aspirate the liquid, add fluorescent secondary antibody, and protect from light.

[0077] (7) Incubate in a wet box at 37°C for 1 hour and rinse the sections three times with PBST for 3 minutes each time.

[0078] (8) Add DAPI and incubate in the dark for 5 minutes. Wash off excess DAPI with PBST for 5 minutes x 4 times.

[0079] (9) Aspirate the liquid, seal the slides with anti-fluorescence quencher sealing solution, and observe under a fluorescence microscope.

[0080] 6. Detection of intracellular ROS levels (1) DHE staining Dihydroethidium reacts with superoxide anions in cells to generate ethidium bromide. Ethidium bromide can bind to RNA or DNA to produce red fluorescence.

[0081] a) Dissolve DHE in DMSO to a 10 mM stock solution and store at -20°C.

[0082] b) Cells were seeded in 12-well plates on glass slides.

[0083] c) Dilute DHE to 10 μM with serum-free medium, add 300 μL to each well, and stain in a 37°C incubator for 30 min.

[0084] d) Wash the cells three times with PBS, remove the coverslip, and place it on a glass slide. The generation and distribution of ROS in living cells were observed under a fluorescence microscope.

[0085] (2) DCFH-DA staining Dichlorofluorescein diacetate DCFH-DA is a lipophilic substance that has no fluorescence itself. ROS in cells oxidizes DCFH-DA into a fluorescent substance DCF.

[0086] a) Prepare cell lysis buffer according to the table below, adjust its pH to 7.5, and store at 4°C.

[0087] Table 3 Cell lysate b) Dissolve DCFH-DA in DMSO to make a 10 mM stock solution. Store at -20°C in the dark.

[0088] c) Dilute the DCFH-DA stock solution to a 10 μM working solution using serum-free medium and mix thoroughly.

[0089] d) Discard the culture medium in the 6-well plate, add 1 mL of DCFH-DA working solution to each well, and incubate at 37°C in the dark for 30 min.

[0090] e) Discard the DCFH-DA working solution, wash three times with pre-cooled PBS in a dark place, add 300 μL of cell lysis solution to each well, and incubate on ice for 10 min.

[0091] f) Scrape the cells and centrifuge at 13000 g at 4°C for 10 min. Take 200 μL of the supernatant and transfer it to a 96-well plate. Use the cell lysate as a blank control, with an excitation light of 485 nm and an emission light of 538 nm.

[0092] g) Each value was quantified by the corresponding porin content.

[0093] 7. Results The results showed that different concentrations of Dictamni cortex (0.1-200 mg / L) had limited effects on HEKa cell viability within 24 hours ( Figure 4 Middle B). When cells were treated with M5, 20 mg / L Dictamni could inhibit excessive cell proliferation ( Figure 4 Middle C). Both Dictamni and NB can inhibit the abnormal proliferation of HEKa cells ( Figure 4 Middle C, Figure 5 Middle B).

[0094] Under M5 treatment, Dictamni and NB could inhibit the phosphorylation of Stat3 and NF-κB ( Figure 4 Middle H and Figure 5 Middle C). After treatment with Dictamni peel, immunofluorescence showed that the fluorescence intensity of P-STAT3 was significantly reduced ( Figure 4 In addition, the mRNA levels of cytokines IL-1α, IL-1β, IL-17A, IL-17F, and TNFα were significantly blocked ( Figure 4 J and Figure 5 Middle F).

[0095] In addition to the anti-inflammatory properties, the present invention also detected the mRNA levels of the antioxidant systems HO-1, NQO-1, and SOD1 / 2. The results showed that ( Figure 4 Middle K), M5 treatment upregulated HO-1, NQO-1 and SOD1 / 2, disrupting the redox balance, and Dictamni peel significantly suppressed the expression levels. Consistent with the results of Dictamni peel, NB treatment also showed antioxidant capacity detected by DHE staining and DCFH-DA ( Figure 5As expected, NB treatment significantly blocked the expression of cytokines IL-1α, IL-1β, IL-6, and TNFα ( Figure 5 Middle F).

[0096] Experimental Example 3 In vivo experiments verify the therapeutic effect of nanoparticles loaded with Dictamni on psoriasis 1. Establishment of IMQ-induced psoriasis mouse model C57BL / 6J mice were randomly divided into 3 groups, 6 mice in each group: Group 1: negative control group; Group 2: model group, marked as IMQ group; Group 3: NB group. Flowchart ( Figure 6 A) shows the experimental process. The mouse back was depilated with an electric clipper and depilatory cream. The depilatory area was 2 cm × 2 cm. After depilation, the psoriasis mouse model was established. The back skin of the mouse was photographed after depilation and before and after treatment. After the treatment, the mouse was killed by cervical dislocation. After taking the back skin, the skin tissue was washed with cold PBS and fixed in 4% paraformaldehyde. The abdominal cavity was opened to take the spleen, which was washed with cold PBS, wiped dry, weighed, and photographed.

[0097] The area and severity index of psoriasis in mice, namely the PASI score, includes three indicators: erythema, infiltration, and epidermal desquamation / squamation. The severity of each item is still scored from 0 to 4 points, 0 = none, 1 = mild, 2 = moderate, 3 = severe, and 4 = extremely severe. The total score is the PASI score, and the photos were taken and the PASI scores of each group of mice were calculated.

[0098] 2. HE staining of mouse tissue (1) Processing of mouse skin specimens The mouse skin tissue, liver tissue, and kidney tissue obtained during the experiment were thoroughly rinsed with physiological saline to remove blood and contaminants, and then the tissue blocks were placed in 4% paraformaldehyde and fixed for 24 hours.

[0099] (2) Slide processing a) After ultrasonic cleaning of the glass slide with detergent, soak it in a mixture of potassium dichromate and concentrated sulfuric acid for 24 hours; b) After taking it out of the acid tank, rinse it with distilled water until it is completely clean, and dry it in an oven at 60°C overnight; c) To prevent tissue from falling off, the slides need to be treated with 3-aminopropyl-3-ethoxysilane. Dilute the 3-aminopropyl-3-ethoxysilane stock solution with acetone at a volume ratio of 1:50 to form a working solution. Place the washed slides in the newly prepared 3-aminopropyl-3-ethoxysilane working solution and leave for 30 seconds before taking them out. d) Take out the slide, wait for 10 seconds, and then put it into acetone solution for 30 seconds to rinse off the unbound 3-aminopropyl-3-ethoxysilane; e) Dry in the oven for 2 hours and pack into boxes for later use.

[0100] (3) Paraffin embedding and tissue sectioning a) Embedding tissue: First, add some liquid paraffin into the mold. After it cools down slightly, place the tissue to be embedded in the paraffin and arrange them neatly. Then, cover the plastic mold box. Finally, add a little liquid paraffin and cool it to make it solid. b) Sectioning: Remove the embedded tissue from the mold, place it on a paraffin slicer, adjust the slice thickness to 4 μm, and slice continuously; c) Bake the slices in a 60°C oven for 90 min and then transfer to a 37°C oven overnight.

[0101] (4) HE staining steps a) Place the sections in a hematoxylin solution for several minutes, and separate the colors in acid and ammonia water for several seconds each; b) Rinse with running water for 1 hour and then put into distilled water for a while; c) Dehydrate in 70% and 90% alcohol for 10 minutes each; d) Stain in alcohol eosin staining solution for 2-3 minutes; e) The sections were dehydrated with ethanol, transparentized with xylene, and mounted with neutral gum.

[0102] (5) Determination of HE staining results, microscopic examination, and image acquisition and analysis.

[0103] 3. Extraction of total protein from mouse skin tissue (1) Pre-cool the tissue grinder, label the grinding tubes and EP tubes, mix RIPA lysis buffer and PMSF in a ratio of 100:1, and place on ice.

[0104] (2) Weigh about 20 mg of tumor tissue into a grinding tube, add 300 μL of lysis buffer, and place 2 grinding beads in each grinding tube.

[0105] (3) Place the mixture in a grinder with the grinding parameters set to 4°C, 60 Hz, and 45 s.

[0106] (4) After the tissue is fully ground, aspirate the supernatant into a new EP tube.

[0107] The subsequent steps are the same as the extraction of cell proteins.

[0108] 4. Extraction of total RNA from mouse skin tissue (1) Cut the mouse back skin tissue to a size of about 0.5 cm × 0.5 cm, add it to a 1.5 mL RNase-Free Eppendorf tube, add 800 μL Trizol lysis buffer, use surgical scissors to cut the tissue into pieces as much as possible, and place it in a -80°C refrigerator for freezing and lysis; (2) Take out some of the tissue and thaw it, grind it with a grinder, add an appropriate amount of Trizol according to the amount of ground tissue, and thaw it on a shaker for 15 minutes; (3) Add 1 / 5 volume of Trizol in chloroform, vortex for 30 seconds, let stand at room temperature for 5 minutes, and centrifuge at 12,000 rpm for 15 minutes at 4°C after the liquid surface is separated; Subsequent steps are the same as above for cell RNA extraction 5. IHC and IF detection of mouse skin tissue (1) Section preparation: After paraffin embedding, the tissue was cut into 4 μm thickness and placed on an adhesive slide.

[0109] (2) Dewaxing and hydration: First, use a slide baking machine to bake at 62°C for 1 h to promote the melting of the paraffin on the slide. Then, soak in fresh xylene I, fresh xylene II, 100%, 100%, 95%, 80%, and 60% ethanol for 10 min in sequence;

[0110] Wash with ultrapure water 3 times, 1 min each time. This step can remove paraffin and organic solvents in the sectioning process.

[0111] (3) Antigen repair: The fixation and sectioning process of the specimen may destroy the spatial conformation of the antigen, and heat treatment is required to restore the immunogenicity of the antigen. After the Tris-EDTA antigen repair solution is heated in a microwave oven until it boils, carefully put the slide in and continue to heat it at medium heat for 15 minutes, waiting for the slide in the antigen repair solution to naturally cool to room temperature. Wash with ultrapure water 3 times, 1 minute each time.

[0112] (4) Block endogenous peroxidase: Use an immunohistochemistry pen to draw a circle around the tissue to prevent liquid extravasation, add the blocking agent and wait for 10 minutes at room temperature; wash with ultrapure water three times, 1 minute each time.

[0113] (5) Blocking: Add PBS solution containing 5% BSA to the tissue and incubate at room temperature for 30 min to prevent nonspecific binding. Shake off without washing.

[0114] (6) Add primary antibody: Add an appropriate amount of primary antibody according to the size of the tissue and incubate overnight at 4°C to promote specific binding reaction. Rinse with PBS buffer 3 times, 3 min each time.

[0115] (7) IHC detection a) Add secondary antibody: Add appropriate amount of secondary antibody to the tissue and incubate at 37℃ for 20 min. Rinse with PBS buffer 3 times, 3 min each time.

[0116] b) Color development: Add freshly prepared DAB color development solution and incubate at room temperature for 90 seconds. Rinse with tap water.

[0117] c) Re-staining: add hematoxylin solution and incubate for 1 min; differentiate, rinse with tap water to turn blue.

[0118] d) Dehydration and transparent mounting: The slides were immersed in gradient ethanol solutions (60%, 80%, 95%, 100%, 100%) for 5 min in sequence for dehydration, and immersed in xylene solution twice for 5 min each time. Afterwards, neutral gum was used for mounting to avoid large bubbles in the tissue.

[0119] e) Result determination: Two pathologists independently observed and interpreted the results under an optical microscope. The staining results were determined based on the staining range and intensity. Staining in the range of <5% was scored as 0 points; staining in the range of 5% to 25% was scored as 1 point; staining in the range of 26% to 50% was scored as 2 points; staining in the range of 51% to 75% was scored as 3 points; staining in the range of >75% was scored as 4 points. Staining intensity score: no staining was scored as 0 points; staining in light yellow was scored as 1 point; staining in brown was scored as 2 points; staining in brown was scored as 3 points. The two were multiplied to obtain the total score. Total score: 0, negative (-); 1~4, weakly positive (+); 5~8, moderately positive (++); 9~12, strongly positive (+++).

[0120] (8) IF detection a) Incubate with secondary antibody: According to the species of primary antibody, add corresponding fluorescent secondary antibody to the tissue surface and incubate at room temperature for 1 hour in the dark. Rinse with PBS buffer 3 times, 5 minutes each time.

[0121] b) Add DAPI dye to the tissue, protect from light, incubate at room temperature for 10 min, and wash the slices three times with PBS buffer, 5 min each time. Note that light should also be protected during washing.

[0122] c) After slightly drying, add anti-fluorescence quenching sealing agent and seal the slides, taking care not to have bubbles.

[0123] d) The sections were observed under a fluorescence microscope and photographed.

[0124] 6. Aha-labeled cell proteome analysis HEKa cells were treated with methionine-free H-DMEM (Sigma, cat# D0422) to minimize the original reserve of methionine, i.e., methionine starvation for 2 hours. Cells were then treated with 4 mM Aha and 4 mM methionine for another 4 hours. After labeling, cells were washed 3 times with 1× PBS and total cell protein was collected. The Click reaction system was as follows: 100 μM tert-butyl 2,2,2-trichloroacetamidoacetate, 2 mM tris(2-carboxyethyl)phosphine, and 2 mM CuSO4. Add and vortex to mix after each addition of reagents. The precipitated and separated proteins were digested into peptides by trypsin. Finally, NeutrAvidin Beads were used to enrich Aha-labeled peptides for mass spectrometry analysis.

[0125] 7. Results In vivo results showed that NB significantly reduced the clinical scores of psoriasis area and severity index ( Figure 6 B-D), and protected mice from IMQ-induced weight loss ( Figure 6 Middle E). NB reduced IMQ-induced epidermal hyperplasia and splenomegaly ( Figure 6 (F~I).

[0126] In addition, since NB is an iron complex, the present invention detected the iron content of the skin sample by a total iron colorimetric assay kit and found that NB increased the iron content ( Figure 6 Regarding the anti-inflammatory and anti-proliferative properties, lower levels of P-STAT3, P-AKT, and PCNA were observed in the IMQ+NB group ( Figure 6 K, L), and lower levels of psoriasis mRNA, IL-23, TNFα ( Figure 6 Middle M).

[0127] Experimental Example 4 Study on the expression genes related to psoriasis 1. Bioinformatics database analysis GEO Database The Gene Expression Omnibus, i.e., the GSE13355, GSE114286, and GSE53541 datasets in the GEO database, was used for the study. Quality control and normalization were performed after the gene expression data were downloaded. All sample data were open access and no ethical approval was required. HSP90AB1 Expression amount.

[0128] 2. Results The sequencing data from GEO databases GSE13355, GSE114286, and GSE53541 show that ( Figure 7 (G, H) Compared with normal controls and UV-treated patients, the lesional skin of psoriasis patients showed HSP90AB1 The expression of β-catenin was significantly increased, and ultraviolet treatment could reduce the expression.

[0129] Based on the above results, the present invention collected skin slices from psoriasis patients and found that compared with the normal population, the epidermis HSP90AB1 Significantly higher.

[0130] Next, the present invention evaluated the protein and mRNA levels of HSP90AB1 in IMQ-stimulated mouse skin and found that the increase ( Figure 7 Medium F and Figure 8 Middle F).

[0131] Experimental Example 5 To investigate the possible targets of NB, the present invention utilized the azido-alanine AHA, an analog of methionine, to characterize the newly synthesized proteins after NB treatment in HEKa cells ( Figure 7 After incubation with AHA to replace methionine, the nascent proteins were labeled and then conjugated to Alkynl-Biotin via click chemistry. They were further detected using streptavidin beads and mass spectrometry enrichment. Several major biological processes were enriched, including protein folding, maintenance of position, cellular response to interleukin 4, and small molecule decomposition processes ( Figure 7 Middle B). Among the 156 proteins that changed significantly after NB treatment, only one protein, HSP90AB1, could overlap and dock with the target proteins of the three chemicals from Dictamni ( Figure 7 (C, D).

[0132] AUY922 is an N-terminus-targeted HSP90 inhibitor with IC values ​​of HSP90α and HSP90β 50 s is 7.8 and 21 nM. Next, the present invention studied the comparative effect of AUY922, an inhibitor of HSP90AB1, on alleviating the IMQ-induced psoriasis phenotype in mice and NB nanoparticles. The specific process is as follows:

[0133] The IMQ model was established using pathogen-free, 8-week-old male C57BL / 6 mice (IMQ modeling method was the same as in Example 3). The mice were randomly divided into 4 groups (n=6 / group): Ctrl control group, IMQ treatment group, NB intervention group and AUY922 intervention group. 30 min before applying IMQ, the NB intervention group (20 mg / kg) and AUY922 (50 mg / kg) intervention group were injected subcutaneously.

[0134] The results show that ( Figure 8 (A-E) Subcutaneous injection of AUY922 can improve the psoriasis phenotype to a certain extent, reduce PASI, and inhibit weight loss and splenomegaly. Skin sections from the back of mice were obtained for H&E staining, immunohistochemistry staining (HSP90AB1 antibody) and immunofluorescence staining (HSP90AB1 and PCNA antibodies). Compared with the NB group, the AUY922 group showed a weak function against IMQ stimulation, see Figure 8 Middle F. Overall, AUY922 did not alter the expression level of HSP90AB1 as much as NB injection, which significantly decreased after NB administration.

[0135] In summary, the therapeutic efficacy of AUY922 is inferior to that of NB.

[0136] Those skilled in the art should be aware that the scope of the present invention is not limited to the technical solutions formed by a specific combination of the technical features described in the present invention, but should also cover other technical solutions formed by any combination of the technical features described in the present invention or their equivalent features.

Claims

1. A Chinese medicine nanoparticle for treating psoriasis, characterized in that: It is prepared according to the following steps: The aqueous solution of Dictamni cortex is mixed with a buffer solution, Pluronic F-127 is added to encapsulate the mixture, and then the mixture is coupled with ferric chloride hexahydrate to obtain the traditional Chinese medicine nanoparticles.

2. The Chinese medicine nanoparticles according to claim 1, characterized in that: The aqueous solution of Dictamni cortex is obtained by uniformly dispersing Dictamni cortex particles in water.

3. The Chinese medicine nanoparticles according to claim 1, characterized in that: The buffer is Tris buffer or 4-hydroxyethylpiperazineethanesulfonic acid or PBS solution or borate solution.

4. The Chinese medicine nanoparticles according to claim 3, characterized in that: After mixing the Tris buffer and the aqueous solution of Dictamni cortex, gradually adding the aqueous solution of ferric chloride hexahydrate and the Pluronic F-127 solution, mixing and dispersing, to obtain the traditional Chinese medicine nanoparticles; The concentration of Tris buffer is 10mM~100mM, the concentration of Dictamni cortex aqueous solution is 5mg / mL~20mg / mL, the concentration of ferric chloride hexahydrate aqueous solution is 5mg / mL~50mg / mL, the mass concentration of Pluronic F-127 solution is 0.1%~5%, and the volume ratio of Tris buffer, Dictamni cortex aqueous solution, ferric chloride hexahydrate aqueous solution and Pluronic F-127 solution is 25:1:1:

4.

5. The Chinese medicine nanoparticles according to claim 1, characterized in that: The dispersion was carried out by ultrasonic treatment at 30% power for 2 to 3 times.

6. The Chinese medicine nanoparticles according to claim 1, characterized in that: The particle size of the traditional Chinese medicine nanoparticles is 34.94-35.58 nm.

7. Use of the traditional Chinese medicine nanoparticles according to claim 1 in preparing a drug for treating psoriasis.

8. A drug for treating psoriasis, characterized in that: The drug contains the traditional Chinese medicine nanoparticles according to claim 1 as the only active ingredient.

9. The drug according to claim 8, wherein the drug is prepared by compounding the traditional Chinese medicine nanoparticles with pharmaceutically acceptable excipients.

10. The drug according to claim 9, which is an oral preparation, an external preparation or an injection preparation.

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