Traditional Chinese medicine nanoparticles for treating psoriasis and their applications

Abstract

The present invention relates to the field of biomedicine, and specifically to a traditional Chinese medicine nanoparticle for treating psoriasis and its application. The nanoparticles are prepared by mixing an aqueous solution of Dictamnus cortex with a buffer solution, adding Pluronic F-127 to coat the mixture, and then coupling the mixture with ferric chloride hexahydrate to obtain the nanoparticles. These nanoparticles have significantly improved water solubility, are well absorbed by keratinocytes and mice, and exhibit a significant therapeutic effect on psoriasis.

Description

Technical Field

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

[0002] Psoriasis is a skin disease associated with immune activation. Its pathological mechanisms involve a complex interplay between the innate and adaptive immune systems. Histologically, it is characterized by excessive proliferation and abnormal differentiation of keratinocytes (KCs), dermal inflammatory infiltration, and angiogenesis. Psoriasis has a high incidence and is increasing annually. Its recurrence rate can reach as high as 98.4%, and it is often accompanied by chronic illness, disfigurement, disability, and related complications, severely impacting and burdening patients' physical and mental health.

[0003] Current treatments for psoriasis mainly include topical medications, phototherapy, systemic therapy, traditional Chinese medicine, and other adjuvant therapies. 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. However, biological agents still have problems such as poor efficacy in some patients, increased risk of infection, 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 dosage and course of treatment must be controlled. Therefore, the key to psoriasis research is to develop targeted drugs to improve patients' health and quality of life in response to the medical needs of psoriasis.

[0004] my country has a history of using traditional Chinese medicine (TCM) to intervene in the progression of psoriasis spanning thousands of years, accumulating extensive experience and developing unique TCM theories for treating psoriasis. Numerous traditional Chinese herbal remedies have been reported to effectively alleviate the development and progression of psoriasis, including by inhibiting the rapid proliferation of keratinocytes and reducing the expression of inflammatory factors. However, unclear targets and inter-batch variability in composition hinder the integration of TCM into the global healthcare system. Dictamnus cortex is a traditional Chinese medicine, documented in the Shennong Bencao Jing (Classic of Materia Medica), and the Chinese Pharmacopoeia lists it in dozens of prescriptions for treating skin diseases. Dictamnus cortex can be taken orally, applied topically, or injected locally. It has been shown to clear heat, detoxify, remove dampness, and alleviate itching in skin diseases. It is widely used to treat tinea pedis, urticaria, eczema, psoriasis, flat warts, and skin cancer. However, its poor water solubility has hampered its development and application. Summary of the Invention

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

[0006] The specific technical solutions provided by the present invention are as follows:

[0007] In a first aspect, the present invention provides a Chinese medicine nanoparticle for treating psoriasis, which is prepared according to the following steps:

[0008] The aqueous solution of Dictamnus cortex is mixed with a buffer solution, Pluronic F-127 is added to coat the mixture, and then the mixture is coupled with ferric chloride hexahydrate to obtain the traditional Chinese medicine nanoparticles.

[0009] The buffer solution is used to adjust the pH value to control the reaction rate and particle stability.

[0010] As a preferred embodiment of the present invention, the aqueous solution of Dictamnus cortex is prepared according to the following steps: Dictamnus cortex particles are uniformly dispersed in water.

[0011] As a preferred embodiment of the present invention, the buffer solution is Tris buffer solution or 4-hydroxyethylpiperazineethanesulfonic acid solution or PBS solution or borate solution.

[0012] As a preferred embodiment of the present invention, after mixing a Tris buffer solution with an aqueous solution of Dictamnus cortex, an aqueous solution of ferric chloride hexahydrate and a Pluronic F-127 solution are gradually added, mixed and dispersed to obtain the traditional Chinese medicine nanoparticles;

[0013] The concentration of Tris buffer is 10 mM~100 mM, the pH is 8.0~9.0, the concentration of the aqueous solution of Dictamnus cortex is 5 mg / mL~20 mg / mL, the concentration of the aqueous solution of ferric chloride hexahydrate is 5 mg / mL~50 mg / mL, the mass concentration of Pluronic F-127 is 0.1%~5%, and the volume ratio of Tris buffer, aqueous solution of Dictamnus cortex, aqueous solution of ferric chloride hexahydrate and Pluronic F-127 solution is 25:1:1:4.

[0014] As a preferred embodiment of the present invention, the dispersion is performed by ultrasonic treatment at 30% power for 2 to 3 times. After ultrasonic treatment, the particles can be evenly distributed and agglomeration and precipitation of the particles can be prevented.

[0015] As a preferred embodiment of the present invention, the particle size of the traditional Chinese medicine nanoparticles is 34.94-35.58 nm.

[0016] The second aspect of the present invention provides a use of the traditional Chinese medicine nanoparticles in preparing a drug for treating psoriasis.

[0017] In a third aspect, the present invention provides a drug for treating psoriasis, wherein the drug contains the traditional Chinese medicine nanoparticles as the sole active ingredient.

[0018] As a preferred embodiment of the present invention, the medicine is prepared by compounding the traditional Chinese medicine nanoparticles with pharmaceutically acceptable excipients.

[0019] As a preferred embodiment of the present invention, the drug is an oral preparation, an external preparation or an injection preparation.

[0020] Compared with the prior art, the beneficial effects of the present invention are:

[0021] (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 placed at room temperature. The addition of Pluronic F-127 greatly enhances the water solubility and bioavailability, and can continue to maintain stability in physiological environments such as PBS and culture medium. At the same time, ferric chloride hexahydrate (hexacoordinated) is used to couple the 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 - It objectively exists in the internal body environment, so the use of ferric chloride hexahydrate will not introduce other elements and is highly safe.

[0022] (2) The Chinese medicine nanoparticles provided by the present invention for the treatment of psoriasis 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 the therapeutic effects and related molecular mechanisms of psoriasis. Based on the functions of this Chinese medicine nanotechnology, its application can be extended to clinical practice.

[0023] (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 psoriatic skin.

[0024] (4) The present invention uses experiments combined with bioinformatics analysis to clarify the mechanism of action of Dictamnus cortex in the treatment of psoriasis. It was verified that after Dictamnus cortex acts on keratinocytes or mice, it causes a decrease in HSP90AB1 and its binding protein CDC37. HSP90AB1 is a subunit of HSP90. HSP90 is widely overexpressed in inflammatory diseases and cancers, and its client proteins are as many 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

[0025] Figure 1 Characterization of Dictamni cortex (DBC)-loaded nanoparticles (NB). A. Changes in the dissolution of Dictamni cortex in water over 72 hours; B. Schematic diagram of NB synthesis; C. TEM image of NB particles; D. Lorenz TEM image (scale = 100 nm, located on the left side of the first horizontal row) and energy dispersive spectrometry (EDS) of NB particles; E-F. Hydrated particle size (E) and zeta potential (F) of NB; G-H. Changes in NB particle size over 7 days in PBS solution (G) and culture medium (H).

[0026] Figure 2 Characterization of Dictamni cortex (NB)-loaded nanoparticles. A. Fourier transform infrared (FTIR) spectrum of NB particles; B. UV spectrum of NB particles; C-D. X-ray photoelectron spectroscopy (XPS) spectra of NB particles, including the Fe2p spectrum (C) and O1s spectrum (D); E. Fe2p spectrum of FeCl3; F. O1s spectrum of Dictamni cortex.

[0027] Figure 3 Characterization of Dictamni cortex (NB)-loaded nanoparticles. A-B. Images (A) and fluorescence spectra (B) of NB particles induced by UV light. C-D. Cumulative release of Dictamni cortex (C) and Fe from NB particles at different time points under different pH conditions (pH = 5.5, 6.8, 7.4). 3+ The data represent the mean ± SEM from at least three independent experiments. ***p < 0.001, **p < 0.01, *p < 0.05.

[0028] Figure 4Dictamnus cortex inhibits M5-induced HEKa cell proliferation and inflammatory response. A. Historical origin of Dictamnus cortex. B. Effects of different concentrations of Dictamni peel treatment for 24 h on HEKa cell viability; C. 20 mg / L Dictamni peel inhibited 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 with IL-6 / TNF; F. Venn diagram analysis of the common molecular targets of luteolin, wogonin, and quercetin; G. Molecular docking models of luteolin, wogonin, and quercetin with IL-6 and TNF; H. Western blotting analysis of p-NF-κB / NF-κB and p-STAT3 / STAT3 in M5- and Dictamni peel-treated groups; I. Immunofluorescence localization of p-STAT3 after Dictamni peel and M5 co-treatment for 24 h (Scale bar = 10 μm); J. Relative mRNA expression levels of inflammatory factors (IL-1α, IL-1β, IL-17A, IL-17F, TNFα); K. Relative mRNA expression levels of antioxidant system (HO-1, NQO-1, SOD1 / 2); 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 analysis of variance with Bonferroni correction.

[0029] Figure 5 NB inhibits abnormal cell proliferation and inflammatory responses. A. Schematic diagram of the experimental flow chart for HEKa cells treated with M5 and NB; B. Quantitative analysis of cell proliferation in different treatment groups at 24 / 48 / 72 h by CCK8 assay; C. Western blot analysis of p-NF-κB / NF-κB and p-STAT3 / STAT3 in the M5 / NB treatment groups; D. DHE fluorescence staining to track intracellular ROS levels; E. DCFH-DA probe assay to detect ROS production; F. Relative mRNA expression levels of inflammatory cytokines IL-1α, IL-1β, IL-6, and TNFα. Data are expressed as mean ± standard error (n = 3 or 6 independent biological samples). *p < 0.05, **p < 0.01, ***p < 0.001, n.s., not statistically significant. Two-tailed t-test / two-way ANOVA with Bonferroni correction.

[0030] Figure 6Subcutaneous administration of NB alleviates psoriatic dermatitis in mice. A. Schematic diagram of the experimental process; B. Typical images of the phenotype of mice in each group on treatment days 0, 3, and 5; C. Fluorescence microscopy of skin tissue in the NB-treated group; D. Psoriasis area and severity index scores; E. Curves of mouse weight changes during the experiment; F. H&E-stained pathological sections of the dorsal skin of mice in each group; 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-to-body ratio (spleen weight / body weight) (normalized to the control group); J. Colorimetric determination of total iron content (OD 593nm K. Immunohistochemical staining of p-STAT3 (first row) and colocalization of PCNA with immunofluorescence (second row) in lesional tissue; L. Western blot analysis of p-Akt / Akt and p-STAT3 / STAT3 in skin tissue; M. Relative mRNA expression of psoriasis-related cytokines (IL-6, IL-1α, IL-1β, IL-17A, IL-17F, IL-22, IL-23, TNFα). Scale bar = 100 μm. Data are expressed as mean ± standard error (n = 6 animals / group). *p < 0.05, **p < 0.01, ***p < 0.001, n.s., not statistically significant. Two-tailed t-test / two-way ANOVA with Bonferroni correction.

[0031] Figure 7NB-triggered nascent proteomic changes identify 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. Heat map (C) and volcano plot (D) of nascent proteins significantly altered after NB treatment compared with the Ctrl group; E. Molecular docking simulations of HSP90AB1 with luteolin, wogonin, and quercetin. The upper row shows the structural schematics of luteolin, wogonin, and quercetin; the lower row shows molecular docking simulations. The right side of each panel shows the magnified image within the corresponding red box on the left; F. Immunoblotting of HSP90AB1 in IMQ-induced mouse skin (vs. control); G. HSP90AB1 expression levels from the GEO database (GSE13355, GSE114286, GSE53431); H. Immunofluorescence staining of HSP90AB1 in skin tissue from healthy donors and patients with psoriasis. The right panel shows the magnified image within the white box on the left. Scale bar = 100 μm. Data are presented as mean ± SEM (n = 3 biologically independent cell samples or 6 mice per group). Mass spectrometry analysis was thresholded at |log2 fold change| ≥ 1 and p value < 0.05. *p < 0.05, **p < 0.01, ***p < 0.001, ns indicates not significant. Statistical analysis was performed using a two-tailed Student's t-test or two-way analysis of variance with a Bonferroni post hoc test.

[0032] Figure 8 The HSP90 inhibitor AUY922 improves IMQ-induced psoriasis in mice. A. Representative images of skin lesions in the four groups of mice on days 0, 3, and 5; B. Changes in mouse body weight and (C) Psoriasis Area and Severity Index (PASI) scores (Control, IMQ, IMQ+NB, and IMQ+AUY922 groups); D. Gross images of the spleen and (E) spleen weight / body weight ratio (normalized to the control group); F. H&E staining (first row), immunohistochemical staining for HSP90AB1 (second row), and immunofluorescence analysis of HSP90AB1 (third row) and PCNA (fourth row) expression levels in skin tissue from the 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 not significant. Statistical analysis was performed using a two-tailed t-test. DETAILED DESCRIPTION

[0033] The present invention will be further described in detail below with reference to 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.

[0034] 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%. 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.

[0035] Current treatments for psoriasis mainly include topical medications, phototherapy, systemic therapy, traditional Chinese medicine, and other adjuvant therapies. 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. However, biological agents still have problems such as poor efficacy in some patients, increased risk of infection, 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 dosage and course of treatment must be controlled. Therefore, the key to psoriasis research is to develop targeted drugs to improve patients' health and quality of life in response to the medical needs of psoriasis.

[0036] Dictamnus cortex is a traditional Chinese medicine that has been shown to clear away heat and toxic substances, as well as to relieve dampness and itching in the treatment of skin diseases. It is widely used to treat tinea pedis, urticaria, eczema, psoriasis, flat warts, and skin cancer. However, Dictamnus cortex's poor water solubility has hampered its development and application.

[0037] Based on this, the present invention provides a Chinese medicine nanoparticle for treating psoriasis, which is prepared according to the following steps:

[0038] The aqueous solution of Dictamnus cortex is mixed with a buffer solution, Pluronic F-127 is added to coat the mixture, and then the mixture is coupled with ferric chloride hexahydrate to obtain the traditional Chinese medicine nanoparticles.

[0039] Pluronic F-127 is a nonionic surfactant polyol that promotes the dissolution of other substances in physiological media. In this invention, it is used to encapsulate water-soluble drug particles derived from Dictamni cortex. Dictamni cortex has poor water solubility and precipitates upon standing at room temperature. The addition of Pluronic F-127 significantly enhances its water solubility and bioavailability, and allows it to maintain stability in physiological environments such as PBS and culture medium. Furthermore, ferric chloride hexahydrate (hexacoordinated) is used to couple the Pluronic F-127 encapsulated with Dictamni cortex. Iron ion levels are reduced in the skin and blood of psoriasis patients. By coupling iron ions, the skin iron content in a psoriasis mouse model can be significantly increased.

[0040] Example 1

[0041] Nanoparticles loaded with Dictamnus cortex are synthesized using Dictamnus cortex (traditional Chinese medicine), Pluronic F-127, ferric chloride hexahydrate, and Tris buffer solution as raw materials, including the following preparation steps (taking 5 ml system as an example): Figure 1 As shown in B:

[0042] (1) Place 5 ml of 10 mM Tris buffer on a magnetic stirrer and add 200 μL of 10 mg / mL Dictamnus cortex aqueous solution while rotating;

[0043] (2) Add 200 μL of 10 mg / mL ferric chloride hexahydrate dropwise;

[0044] (3) Add 200 μL of 2% Pluronic F-127 dropwise;

[0045] (4) 30% power ultrasound 3 times;

[0046] (5) Continue stirring for 1 to 2 days.

[0047] The Dictamnus cortex aqueous solution was prepared as follows: 50 mg of Dictamnus cortex particles were weighed using an analytical balance, 5 mL of ddH2O was added, and the mixture was thoroughly shaken to uniformly disperse in water. Figure 1 As shown in A.

[0048] Afterwards, dialyze using a MWCO 3500 ultrafiltration membrane or ultrafiltration tubing to a residual volume of approximately 1 mL. After sonication, store the solution. For use, dissolve it in a 0.9% NaCl aqueous solution and filter it through a 0.45 μm pore size filter membrane. After filtration, it can be directly injected locally. The resulting Dictamnus cortex-loaded nanoparticles are labeled NB particles.

[0049] Example 2

[0050] Nanoparticles loaded with Dictamnus cortex are prepared according to the following steps:

[0051] (1) Place 5 ml of 10 mM Tris buffer on a magnetic stirrer and add 200 μL of 5 mg / mL Dictamnus cortex aqueous solution while rotating;

[0052] (2) Add 200 μL of 50 mg / mL ferric chloride hexahydrate dropwise;

[0053] (3) Add 200 μL of 0.1% Pluronic F-127 dropwise;

[0054] (4) 30% power ultrasound 3 times;

[0055] (5) Continue stirring for 1 to 2 days.

[0056] The Dictamnus cortex aqueous solution was prepared according to the following steps: 50 mg of Dictamnus cortex granules were weighed using an analytical balance, 5 mL of ddH2O was added, and the mixture was fully shaken to be uniformly dispersed in water.

[0057] Afterwards, dialyze using a MWCO 3500 ultrafiltration membrane or ultrafiltration tubing to a residual volume of approximately 1 mL. The solution is then stored after sonication. Upon use, it is dissolved in a 0.9% NaCl aqueous solution and filtered through a 0.45 μm pore size filter membrane. After filtration, it can be directly injected locally. The resulting Dictamnus cortex-loaded nanoparticles are labeled NB particles.

[0058] Example 3

[0059] Nanoparticles loaded with Dictamnus cortex are prepared according to the following steps:

[0060] (1) Place 5 ml of 20 mM Tris buffer on a magnetic stirrer and add 200 μL of 10 mg / mL Dictamnus cortex aqueous solution while rotating;

[0061] (2) Add 200 μL of 50 mg / mL ferric chloride hexahydrate dropwise;

[0062] (3) Add 200 μL of 5% Pluronic F-127 dropwise;

[0063] (4) 30% power ultrasound 3 times;

[0064] (5) Continue stirring for 1 to 2 days.

[0065] The Dictamnus cortex aqueous solution was prepared according to the following steps: 50 mg of Dictamnus cortex granules were weighed using an analytical balance, 5 mL of ddH2O was added, and the mixture was fully shaken to be uniformly dispersed in water.

[0066] Afterwards, dialyze using a MWCO 3500 ultrafiltration membrane or ultrafiltration tubing to a residual volume of approximately 1 mL. The solution is then stored after sonication. Upon use, it is dissolved in a 0.9% NaCl aqueous solution and filtered through a 0.45 μm pore size filter membrane. After filtration, it can be directly injected locally. The resulting Dictamnus cortex-loaded nanoparticles are labeled NB particles.

[0067] Example 4

[0068] Nanoparticles loaded with Dictamnus cortex are prepared according to the following steps:

[0069] (1) Place 5 ml of 50 mM Tris buffer on a magnetic stirrer and add 200 μL of 10 mg / mL Dictamnus cortex aqueous solution while rotating;

[0070] (2) Add 200 μL of 25 mg / mL ferric chloride hexahydrate dropwise;

[0071] (3) Add 200 μL of 2% Pluronic F-127 dropwise;

[0072] (4) 30% power ultrasound 3 times;

[0073] (5) Continue stirring for 1 to 2 days.

[0074] The Dictamnus cortex aqueous solution was prepared according to the following steps: 50 mg of Dictamnus cortex granules were weighed using an analytical balance, 5 mL of ddH2O was added, and the mixture was fully shaken to be uniformly dispersed in water.

[0075] Afterwards, dialyze using a MWCO 3500 ultrafiltration membrane or ultrafiltration tubing to a residual volume of approximately 1 mL. The solution is then stored after sonication. Upon use, it is dissolved in a 0.9% NaCl aqueous solution and filtered through a 0.45 μm pore size filter membrane. After filtration, it can be directly injected locally. The resulting Dictamnus cortex-loaded nanoparticles are labeled NB particles.

[0076] 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.

[0077] TEM images ( Figure 1 The results of C) and dynamic light scattering (DLS) showed that the NB particles were spherical or nearly spherical, with relatively uniform distribution and no obvious agglomeration. The NB particles were 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 well-crystalline nanoparticles. EDS spectrum ( Figure 1 D) shows that the main elements are Fe and O, and the Fe 2+ / Fe 3+ The mixed valence states are consistent. No impurity elements such as C and Si were detected, indicating that the sample is of high purity.

[0078] The hydrated particle size and Zeta potential of NB and Dictamnus cortex were further analyzed ( Figure 1 DLS further proves that BXP and Fe 3+The zeta potentials of NB were -19.5 mV and 10.3 mV, respectively. After the cations were coordinated with BXP, NB exhibited a narrower distribution at -11.8 mV, indicating good dispersion of NB in ​​the solution and no significant aggregation, consistent with TEM observations. Furthermore, Lorenz TEM and energy dispersive spectroscopy (EDS) results demonstrated uniform distribution of carbon (C), oxygen (O), and iron (Fe) within the NB.

[0079] During the cell culture process, the particle size of NB in ​​PBS and DMEM culture medium was detected, and the results showed that NB could remain stable for 7 consecutive days ( Figure 1 (G~H).

[0080] Compared with Dictamnus cortex 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 presence of BXP in NB was confirmed by Fourier transform infrared spectroscopy (FT-IR). Figure 2 A in the middle). For example, 2883 cm -1 The peak at indicates that the proton is absorbed by Fe 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 in 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 vibrations (e.g., carbonate or carboxylate groups), but the peak intensity is weak, possibly related to trace organic contamination or precursor residues on the surface. The absence of the Oh-CO peak associated with 332.5 eV (combined with XPS data) suggests that the surface modification is primarily hydroxyl groups, with no significant organic ligand coverage.

[0081] Ultraviolet-visible (UV-Vis) absorption spectroscopy confirmed the presence of BXP in NB ( Figure 2 B), and the Dictamnus cortex itself has strong UV-visible absorption, which will cause signal interference in biological effect experiments. The UV-visible absorption is reduced through NB assembly and can be maintained stable.

[0082] In addition, X-ray photoelectron spectroscopy (XPS) was used to analyze the energy level changes of iron in NB and Dictamnus cortex. Fe2p is the most common Auger spectrum peak of iron. In addition to the main peak, there is also a half peak (satellite). 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 in the middle). 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 2 The high-resolution O 1S spectrum of NB shows characteristic peaks at 533.52 eV and 532.13 eV ( Figure 2 In the middle (D), the high-resolution O 1S spectrum of BXP shows characteristic peaks at 530.98 eV and 532.73 eV ( Figure 2 F in the middle). This indicates that Fe + The electron pair of O atoms is shared with BXP, and the O atoms are closely related 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).

[0083] Dictamnus peel and NB can produce autofluorescence after ultraviolet excitation, which may be caused by some compounds in Dictamnus peel. The fluorescence spectrum detected the maximum excitation wavelength (Ex) of Dictamnus peel and NB at 405 nm and the maximum emission wavelength (Em) at 468 nm, see Figure 3 Middle A~B.

[0084] The release of Dictamnus chinensis and Fe 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 Dictamnus and Fe 3+ The release of NB was significantly enhanced, with more than 80% of the drug released in the same time, which was much higher than that 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.

[0085] Experimental Example 1

[0086] Bioinformatics analysis

[0087] (1) Molecular docking

[0088] The molecular structures of woflavin, luteolin, and quercetin, the core components of Dictamnus cortex, were obtained from the PubChem database (https: / / pubchem.ncbi.nlm.nih.gov / ) and energy minimized using Chem3D software.

[0089] 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).

[0090] 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, the docking results were visualized using PYMOL software.

[0091] (2) Cluster analysis

[0092] 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.

[0093] Through the analysis of multiple component targets of Dictamnus cortex, the most significant changes in several GO biological processes were summarized ( Figure 4 Middle D) and key change pathways ( Figure 4 The common molecular targets of the three key ingredients, luteolin, wogonin, and quercetin, were further analyzed using a Venn diagram ( Figure 4 F in the middle). Molecular docking revealed that luteolin, wogonin, and quercetin can all dock with IL-6 and TNF ( Figure 4 Middle G).

[0094] Experimental Example 2

[0095] Cell experiments verify the therapeutic effect of nanoparticles loaded with Dictamnus cortex on psoriasis

[0096] 1. Cell culture and treatment

[0097] HEKa cells were cultured in DMEM complete medium supplemented with 10% fetal bovine serum (10% by volume) at 37°C in a 5% CO2 incubator. Psoriatic cellular inflammation was simulated using 10 ng / ml M5, which contains IL-1α, IL-17A, IL-22, oncostatin M, and tumor necrosis factor-α.

[0098] 2. CCK-8 cell viability assay

[0099] The experimental process is shown in Figure 5 Middle A.

[0100] (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.

[0101] (2) Drug treatment: discard the original culture medium and add fresh culture medium containing different concentrations of Dictamni or NB nanoparticles. Set up 6 replicate wells in each group and treat for 24, 48, and 72 h, respectively.

[0102] (3) Color development reaction: Add 10 μL of CCK-8 solution to each well and incubate in the dark for 2 h.

[0103] (4) Absorbance detection: The absorbance at 450 nm was measured by an enzyme-linked microplate reader with a reference wavelength of 650 nm, and the background value of the blank well was subtracted.

[0104] (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.

[0105] 3. Total RNA extraction, reverse transcription and qRT-PCR from cell tissues

[0106] Total RNA was extracted from cell tissues and reverse transcribed. The reverse transcribed products were then subjected to PCR to detect specific gene expression. The reverse transcription system is shown in Table 1, and the PCR reaction system is shown in Table 2.

[0107] Table 1 Reverse transcription system

[0108]

[0109] Reverse transcription conditions were: 25°C, 10 min; 42°C, 30 min; 85°C, 5 min;

[0110] Table 2 PCR system

[0111]

[0112] PCR reaction conditions are as follows:

[0113]

[0114] 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.

[0115] 4. Total cell protein extraction and Western blot detection

[0116] Total protein was extracted by IP lysis, followed by BCA quantification and protein concentration adjustment. Western blot analysis was then performed using primary and secondary antibodies against p-NF-κB / NF-κB and p-STAT3 / STAT3. Finally, ECL luminescence was used to detect the grayscale values ​​of protein bands, and the results were quantitatively analyzed.

[0117] 5. Cell Immunofluorescence Staining

[0118] (1) Inoculate 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 for 3 minutes each time.

[0119] (2) Fix with 4% paraformaldehyde and rinse with PBS three times, 3 minutes each time.

[0120] (3) 0.5% Triton X-100, permeabilization for 20 minutes.

[0121] (4) Wash with PBS three times, 3 minutes each time, aspirate the PBS, add goat serum, and block for 30 minutes.

[0122] (5) Aspirate the blocking solution, add primary antibody without washing, and incubate at 4°C overnight.

[0123] (6) Wash with PBST three times, 3 minutes each time, aspirate the liquid, add fluorescent secondary antibody, and protect from light.

[0124] (7) Incubate in a wet box at 37°C for 1 hour, and then rinse the sections with PBST three times for 3 minutes each time.

[0125] (8) Add DAPI and incubate in the dark for 5 minutes. Wash off excess DAPI with PBST for 5 minutes x 4 times.

[0126] (9) Aspirate the liquid, seal the slides with anti-fluorescence quencher sealing solution, and observe under a fluorescence microscope.

[0127] 6. Detection of intracellular ROS levels

[0128] (1) DHE staining

[0129] Dihydroethidium reacts with superoxide anions in cells, dehydrogenating to form ethidium bromide. Ethidium bromide can bind to RNA or DNA, producing red fluorescence.

[0130] a) Dissolve DHE in DMSO to a 10 mM stock solution and store at -20°C.

[0131] b) Cells were seeded in 12-well plates on glass slides.

[0132] c) Dilute DHE to 10 μM in serum-free medium, add 300 μL to each well, and stain in a 37°C incubator for 30 min.

[0133] d) The slides were washed three times with PBS, and the coverslips were removed and placed on glass slides. The generation and distribution of ROS in living cells were observed under a fluorescence microscope.

[0134] (2) DCFH-DA staining

[0135] Dichlorofluorescein diacetate (DCFH-DA) is a lipophilic substance that is non-fluorescent. Intracellular ROS oxidizes DCFH-DA into the fluorescent substance DCF.

[0136] a) Prepare cell lysis buffer according to the table below, adjust the pH to 7.5, and store at 4°C.

[0137] Table 3 Cell lysate

[0138]

[0139] b) Dissolve DCFH-DA in DMSO to make a 10 mM stock solution. Store at -20°C in the dark.

[0140] c) Dilute the DCFH-DA stock solution to a 10 μM working solution in serum-free medium and mix thoroughly.

[0141] 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.

[0142] e) Discard the DCFH-DA working solution, wash three times with pre-chilled PBS in the dark, add 300 μL of cell lysis buffer to each well, and incubate on ice for 10 min.

[0143] f) Scrape the cells and centrifuge at 13,000 g at 4°C for 10 min. Transfer 200 μL of the supernatant to a 96-well plate. Use the cell lysate as a blank control. Excitation is at 485 nm, and emission is at 538 nm.

[0144] g) Each value was quantified by the corresponding porin content.

[0145] 7. Results

[0146] The results showed that different concentrations of Dictamnus 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 Dictamnus cortex could inhibit the excessive proliferation of cells ( Figure 4 Middle C). Both Dictamni and NB can inhibit the abnormal proliferation of HEKa cells ( Figure 4 Middle C, Figure 5 Middle B).

[0147] Under M5 treatment, Dictamnus cortex 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).

[0148] In addition to its 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, while Dictamnus edulis significantly suppressed the expression levels. Consistent with the results of Dictamnus edulis, NB treatment also showed antioxidant capacity detected by DHE staining and DCFH-DA ( Figure 5 As expected, NB treatment significantly blocked the expression of cytokines IL-1α, IL-1β, IL-6, and TNFα ( Figure 5 Middle F).

[0149] Experimental Example 3

[0150] In vivo experiments verify the therapeutic effect of Dictamnus cortex-loaded nanoparticles on psoriasis

[0151] 1. Establishment of IMQ-induced psoriasis mouse model

[0152] C57BL / 6J mice were randomly divided into 3 groups, with 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 Center (A) shows the experimental procedure. Hair was removed from the back of mice using electric clippers and depilatory cream, covering an area of ​​2 cm x 2 cm. A psoriasis mouse model was established after depilation. Photos of the mouse back skin were taken after depilation and before and after treatment. After treatment, the mice were sacrificed by cervical dislocation. The back skin was removed, washed with cold PBS, and fixed in 4% paraformaldehyde. The spleen was opened and removed, washed with cold PBS, dried, weighed, and photographed.

[0153] The mouse psoriasis lesion area and severity index (PASI) score was used to assess the severity of psoriasis. The PASI score includes three indicators: erythema, infiltration, and epidermal scaling / scaling. The severity of each indicator is still rated on a scale of 0 to 4, with 0 = none, 1 = mild, 2 = moderate, 3 = severe, and 4 = extremely severe. The total score is the PASI score. Photos were taken and the PASI score was calculated for each group of mice.

[0154] 2. HE staining of mouse tissue

[0155] (1) Processing of mouse skin specimens

[0156] 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 h.

[0157] (2) Slide processing

[0158] 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;

[0159] b) After removing from the acid tank, rinse with distilled water until completely clean, and then dry in an oven at 60°C overnight;

[0160] c) To prevent tissue from falling off, 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 make a working solution. Place the washed slides in the newly prepared 3-aminopropyl-3-ethoxysilane working solution for 30 seconds before removing.

[0161] d) Remove the slide, wait for 10 seconds, and then place it in acetone solution for 30 seconds to rinse off unbound 3-aminopropyl-3-ethoxysilane;

[0162] e) Dry in the oven for 2 hours, then pack in boxes and set aside.

[0163] (3) Paraffin embedding and tissue sectioning

[0164] a) Embedding tissue: First, add some liquid paraffin to the mold. After it cools slightly, place the tissue to be embedded in the paraffin and arrange it neatly. Then, close the plastic mold box. Finally, add a small amount of liquid paraffin and cool it until it becomes solid.

[0165] b) Sectioning: Remove the embedded tissue from the mold and place it on a paraffin microtome. Adjust the slice thickness to 4 μm and slice continuously.

[0166] c) Bake the slices in a 60°C oven for 90 min, then transfer to a 37°C oven overnight.

[0167] (4) HE staining steps

[0168] a) Place the sections in a hematoxylin solution for several minutes, and separate the colors in acid and ammonia water for several seconds each.

[0169] b) Rinse with running water for 1 hour and then put into distilled water for a while;

[0170] c) Dehydrate in 70% and 90% alcohol for 10 minutes each;

[0171] d) Stain in alcohol eosin solution for 2-3 minutes;

[0172] e) The sections were dehydrated with ethanol, cleared with xylene, and mounted with neutral gum.

[0173] (5) Determination of HE staining results, microscopic examination, and image acquisition and analysis.

[0174] 3. Extraction of total protein from mouse skin tissue

[0175] (1) Pre-cool the tissue grinder, label the grinding tube and EP tube, mix RIPA lysis buffer and PMSF at a ratio of 100:1, and place on ice.

[0176] (2) Weigh about 20 mg of tumor tissue and place it in a grinding tube. Add 300 μL of lysis buffer and place 2 grinding beads in each grinding tube.

[0177] (3) Place in a grinder with the grinding parameters of 4°C, 60 Hz, and 45 s.

[0178] (4) After the tissue is fully ground, aspirate the supernatant into a new EP tube.

[0179] The subsequent steps are the same as those for cell protein extraction.

[0180] 4. Extraction of total RNA from mouse skin tissue

[0181] (1) Cut the mouse back skin tissue to a size of approximately 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 small pieces as much as possible, and place it in a -80°C refrigerator for freezing and lysis;

[0182] (2) After thawing, remove the tissue and grind it in a grinder. Add an appropriate amount of Trizol according to the amount of ground tissue and thaw it on a shaker for 15 minutes.

[0183] (3) Add 1 / 5 volume of Trizol-based 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 separates.

[0184] Subsequent steps are the same as above for cell RNA extraction

[0185] 5. IHC and IF detection of mouse skin tissue

[0186] (1) Section preparation: The tissue was embedded in paraffin and cut into 4 μm thickness and placed on an adhesive slide.

[0187] (2) Dewaxing and hydration: First, use a slide baker at 62°C for 1 hour 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 minutes in sequence;

[0188] Wash with ultrapure water three times, 1 min each time. This step can remove paraffin and organic solvents used in the sectioning process.

[0189] (3) Antigen retrieval: The specimen fixation and sectioning process may destroy the spatial conformation of the antigen, requiring heat treatment to restore the immunogenicity of the antigen. Heat the Tris-EDTA antigen retrieval solution in a microwave oven until it boils. Then carefully place the slide in the microwave oven and continue heating on medium heat for 15 minutes. Allow the slide in the antigen retrieval solution to cool to room temperature. Rinse with ultrapure water three times, 1 minute each time.

[0190] (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.

[0191] (5) Blocking: Add 5% BSA in PBS to the tissue and incubate at room temperature for 30 min to prevent nonspecific binding. Shake off without washing.

[0192] (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 three times, 3 minutes each time.

[0193] (7) IHC testing

[0194] a) Add secondary antibody: Add an appropriate amount of secondary antibody to the tissue and incubate at 37°C for 20 min. Rinse three times with PBS buffer, each time for 3 min.

[0195] b) Color development: Add freshly prepared DAB color development solution and incubate at room temperature for 90 seconds. Rinse with tap water.

[0196] c) Counterstaining: Add hematoxylin solution and incubate for 1 min. After differentiation, rinse with tap water to restore the blue color.

[0197] d) Dehydration and transparent mounting: Dehydrate the slides by soaking them in a gradient ethanol solution (60%, 80%, 95%, 100%, 100%) for 5 minutes each. Then, soak them in xylene twice for 5 minutes each. Then, mount the slides with neutral gum to avoid large bubbles in the tissue.

[0198] e) Result Interpretation: Two pathologists independently observed and interpreted the results under a light microscope. Staining results were assessed based on both the extent and intensity of staining. Staining within <5% was assigned a score of 0; staining within 5% to 25% was assigned a score of 1; staining within 26% to 50% was assigned a score of 2; staining within 51% to 75% was assigned a score of 3; and staining within >75% was assigned a score of 4. Staining intensity was scored as follows: no staining was assigned a score of 0; light yellow staining was assigned a score of 1; brownish yellow staining was assigned a score of 2; and brownish brown staining was assigned a score of 3. The total score was calculated by multiplying the two scores. The total score was: 0, negative (-); 1-4, weakly positive (+); 5-8, moderately positive (++); and 9-12, strongly positive (+++).

[0199] (8) IF detection

[0200] a) Incubation with secondary antibodies: Depending on the species of the primary antibody, add the corresponding fluorescent secondary antibody to the tissue surface and incubate at room temperature in the dark for 1 hour. Rinse three times with PBS buffer, 5 minutes each time.

[0201] b) Add DAPI dye to the tissue, protect from light, and incubate at room temperature for 10 minutes. Wash the sections three times with PBS buffer, 5 minutes each time, also protecting from light during washing.

[0202] c) After slightly drying, add anti-fluorescence quenching mounting medium and seal the slides, taking care to avoid air bubbles.

[0203] d) The sections were observed under a fluorescence microscope and photographed.

[0204] 6. Aha-labeled cell proteome analysis

[0205] HEKa cells were treated with methionine-free H-DMEM (Sigma, cat# D0422) to minimize their initial methionine reserves, i.e., methionine starvation for 2 hours. Cells were then treated with 4 mM Aha and 4 mM methionine for an additional 4 hours. After labeling, cells were washed three times with 1× PBS, and total cellular protein was collected. The Click reaction system was as follows: 100 μM tert-butyl 2,2,2-trichloroacetamidate, 2 mM tris(2-carboxyethyl)phosphine, and 2 mM CuSO₄. Mixing was performed by vortexing after each reagent addition. The precipitated and isolated proteins were digested with trypsin to generate peptides. Finally, Aha-labeled peptides were enriched using NeutrAvidin Beads for mass spectrometry analysis.

[0206] 7. Results

[0207] 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 Chinese F~I).

[0208] In addition, since NB is an iron complex, the present invention detected the iron content of the skin samples using 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).

[0209] Experimental Example 4

[0210] Study on expressed genes related to psoriasis

[0211] 1. Bioinformatics database analysis

[0212] GEO Database

[0213] The study was conducted using the Gene Expression Omnibus datasets GSE13355, GSE114286, and GSE53541 from the GEO database. Quality control and normalization were performed after downloading the gene expression data. All sample data were open access and no ethical approval was required. HSP90AB1 Expression level.

[0214] 2. Results

[0215] The sequencing data from GEO databases GSE13355, GSE114286, and GSE53541 showed 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.

[0216] 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.

[0217] Next, the present invention evaluated the protein and mRNA levels of HSP90AB1 in IMQ-stimulated mouse skin and found that the expression of HSP90AB1 in the skin of mice was increased ( Figure 7 Medium F and Figure 8 Middle F).

[0218] Experimental Example 5

[0219] To investigate the possible targets of NB, the present invention utilized the methionine analogue azido-alanine (AHA) to characterize the newly synthesized proteins after NB treatment in HEKa cells ( Figure 7 After incubation with AHA to replace methionine, the nascent protein was labeled and then conjugated to Alkynl-Biotin via click chemistry. Further detection was performed using streptavidin beads and mass spectrometry enrichment. Several major biological processes were enriched, including protein folding, position maintenance, cellular response to interleukin-4, and small molecule catabolism ( 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 Dictamnus cortex ( Figure 7 (C, D).

[0220] AUY922 is an N-terminus-targeted HSP90 inhibitor with IC values ​​of HSP90α and HSP90β. 50 s were 7.8 and 21 nM. Next, the present invention studied the comparative effects of the HSP90AB1 inhibitor AUY922 and NB nanoparticles in alleviating IMQ-induced psoriasis phenotype in mice. The specific process is as follows:

[0221] An IMQ model was established using pathogen-free, 8-week-old male C57BL / 6 mice (IMQ modeling was performed as described in Example 3). Mice were randomly divided into four groups (n = 6 / group): a Ctrl control group, an IMQ-treated group, a NB-treated group, and an AUY922-treated group. Thirty minutes prior to IMQ application, the NB-treated group (20 mg / kg) and the AUY922-treated group (50 mg / kg) were subcutaneously injected.

[0222] The results show that ( Figure 8 (A-E) Subcutaneous injection of AUY922 can improve the psoriatic phenotype to some extent, reduce PASI, and inhibit weight loss and splenomegaly. Skin sections from the back of mice were obtained and subjected to H&E staining, immunohistochemistry (HSP90AB1 antibody), and immunofluorescence staining (HSP90AB1 and PCNA antibodies). Compared with the NB group, the AUY922 group showed weak resistance to IMQ stimulation, as shown in Figure 2. Figure 8 Middle F. Overall, AUY922 did not alter the expression level of HSP90AB1 as observed after NB injection, which significantly decreased HSP90AB1.

[0223] In summary, the therapeutic efficacy of AUY922 is inferior to that of NB.

[0224] 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 Dictamnus cortex is mixed with a buffer solution, Pluronic F-127 is added to coat the mixture, and then the mixture is coupled with ferric chloride hexahydrate to obtain the traditional Chinese medicine nanoparticles; The aqueous solution of Dictamnus cortex is obtained by uniformly dispersing Dictamnus cortex particles in water; after mixing Tris buffer with the aqueous solution of Dictamnus cortex, gradually adding ferric chloride hexahydrate aqueous solution and Pluronic F-127 solution, and ultrasonically treating 2 to 3 times at 30% power; the concentration of Tris buffer is 10 mM to 100 mM, the concentration of the aqueous solution of Dictamnus cortex is 5 mg / mL to 20 mg / mL, the concentration of the aqueous solution of ferric chloride hexahydrate is 5 mg / mL to 50 mg / mL, the mass concentration of Pluronic F-127 solution is 0.1% to 5%, and the volume ratio of Tris buffer, the aqueous solution of Dictamnus cortex, the aqueous solution of ferric chloride hexahydrate and the Pluronic F-127 solution is 25:1:1:4; The particle size of the traditional Chinese medicine nanoparticles is 34.94-35.58 nm.

2. Use of the traditional Chinese medicine nanoparticles according to claim 1 in the preparation of a medicament for treating psoriasis.

3. 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.

4. The drug according to claim 3, wherein the drug is prepared by compounding the traditional Chinese medicine nanoparticles with pharmaceutically acceptable excipients. The drug according to claim 4 , which is an injection preparation.

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