Paclitaxel-loaded nanofiber sponges for the treatment of inflammatory diseases

Abstract

The present application relates to the technical field of medicine, in particular to a paeonia lactiflora extracellular vesicle nano-preparation for treating inflammatory diseases, which active ingredient is an extracellular vesicle extracted from paeonia lactiflora tissues.The paeonia lactiflora extracellular vesicle nano-preparation for treating inflammatory diseases, the present application uses paeonia lactiflora, a traditional medicinal plant, as the source of extracellular vesicles, successfully prepares a nano-preparation with clear activity, compared with the vesicles from animal or human cells which are currently studied more, the plant source vesicles completely avoid the risk of animal source pathogen transmission and potential immune rejection.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a white peony extracellular vesicle nanoformation for the treatment of inflammatory diseases. Background Technology

[0002] White peony is a traditional Chinese medicine with anti-inflammatory and immunomodulatory effects. However, its active ingredients are mostly small molecule compounds with a single mechanism of action. Inflammatory diseases, such as inflammatory bowel disease, are chronic diseases caused by abnormal activation of the immune system.

[0003] Currently, commonly used clinical drugs such as corticosteroids and immunosuppressants have problems such as large side effects and easy relapse. Moreover, there are no reports of using extracellular vesicles derived from white peony for the treatment of inflammatory diseases.

[0004] To address the aforementioned issues, we have made improvements by proposing a peony extracellular vesicle nanoformation for the treatment of inflammatory diseases. Summary of the Invention

[0005] To solve the above-mentioned technical problems, the present invention provides the following technical solution:

[0006] This invention provides a white peony extracellular vesicle nanoformation for the treatment of inflammatory diseases, the active ingredient of which is extracellular vesicles extracted from white peony tissue.

[0007] As a preferred embodiment of the present invention, the extracellular vesicles have a particle size of 50-200 nm.

[0008] As a preferred embodiment of the present invention, the extracellular vesicles can promote the differentiation of regulatory T cells and inhibit the production of inflammatory cytokines IFN-γ, IL-17A and TNF-α.

[0009] A method for preparing extracellular vesicle nanoparticles of Paeonia lactiflora includes the following steps: homogenizing Paeonia lactiflora tissue, removing impurities by differential centrifugation, and enriching and purifying extracellular vesicles by ultracentrifugation or chromatography.

[0010] Application of Paeonia lactiflora extracellular vesicle nanoforms in the preparation of drugs for the prevention or treatment of inflammatory diseases.

[0011] In a preferred embodiment of the present invention, the inflammatory disease is inflammatory bowel disease.

[0012] The beneficial effects of this invention are:

[0013] A nano-formulation of peony extracellular vesicles for the treatment of inflammatory diseases. This invention uses peony, a traditional medicinal plant, as the source of extracellular vesicles and successfully prepares a nano-formulation with definite activity. Compared with the vesicles derived from animal or human cells that are currently the most studied, plant-derived vesicles completely avoid the risk of transmission of animal-derived pathogens and potential immune rejection reactions.

[0014] A nano-formulation of peony extracellular vesicles for the treatment of inflammatory diseases. Peony extracellular vesicles are not simply carriers, but possess strong biological activity. They can simultaneously upregulate the proportion of regulatory T cells and significantly inhibit the production of multiple key pro-inflammatory cytokines. They can more comprehensively and fundamentally correct immune imbalances, potentially leading to more lasting therapeutic effects and reducing the risk of drug resistance. Attached Figure Description

[0015] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used in conjunction with embodiments of the invention to explain the invention and do not constitute a limitation thereof. In the drawings:

[0016] Figure 1 This invention relates to a flow cytometry analysis of CD4 levels in a peony extracellular vesicle nanoparticle formulation for the treatment of inflammatory diseases, after treatment with peony extracellular vesicles. + Foxp3 in T cells + Comparison chart;

[0017] Figure 2 This invention relates to a flow cytometry analysis of CD4 levels in a peony extracellular vesicle nanoparticle formulation for the treatment of inflammatory diseases, after treatment with peony extracellular vesicles. + IFN-γ in T cells + Comparison chart;

[0018] Figure 3 This invention relates to a flow cytometry analysis of CD4 levels in a peony extracellular vesicle nanoparticle formulation for the treatment of inflammatory diseases, after treatment with peony extracellular vesicles. + IL-17A in T cells + Comparison chart;

[0019] Figure 4 This invention relates to a flow cytometry analysis of CD4 levels in a peony extracellular vesicle nanoparticle formulation for the treatment of inflammatory diseases, after treatment with peony extracellular vesicles. + TNF-α in T cells + Comparison chart;

[0020] Figure 5 This is a comparison chart of DAI scores in colitis mice using a white peony extracellular vesicle nanoformation for the treatment of inflammatory diseases according to the present invention.

[0021] Figure 6This is a comparative image of the colorectal length in mice of a white peony extracellular vesicle nanoformation for the treatment of inflammatory diseases according to the present invention. Detailed Implementation

[0022] The preferred embodiments of the present invention will be described below with reference to the accompanying drawings. It should be understood that the preferred embodiments described herein are for illustration and explanation only and are not intended to limit the present invention.

[0023] Example 1: As Figures 1-6 As shown, a peony extracellular vesicle nanoparticle formulation for the treatment of inflammatory diseases has an active ingredient that is an extracellular vesicle extracted from peony tissue.

[0024] Furthermore, the extracellular vesicles have a particle size of 50-200 nm.

[0025] Furthermore, the extracellular vesicles can promote the differentiation of regulatory T cells and inhibit the production of inflammatory cytokines IFN-γ, IL-17A and TNF-α.

[0026] Mouse spleen cells were isolated, activated by stimulation with anti-CD3 / CD28 antibody, and co-cultured with Paeonia lactiflora EVs for 48 hours. Flow cytometry analysis showed that, compared with the control group, the Foxp3 level in the Paeonia lactiflora EVs-treated group was significantly higher. + CD4 + The proportion of T cells increased significantly, while IFN-γ + IL-17A + TNF-α + CD4 + The proportion of T cells decreased significantly.

[0027]

[0028] Table 1. Data on in vitro immunomodulatory activity validation

[0029] Example 2: A method for preparing extracellular vesicle nanoparticles of Paeonia lactiflora, comprising the following steps: homogenizing Paeonia lactiflora tissue, removing impurities by differential centrifugation, and enriching and purifying extracellular vesicles by ultracentrifugation or chromatography;

[0030] Material selection and pretreatment: Fresh or frozen white peony tissue was taken, washed, and homogenized at low temperature;

[0031] The coarse extraction and homogenization are centrifuged at low speed to remove tissue fragments, and the supernatant is then centrifuged at high speed (e.g., 10,000-20,000g) to remove large particulate impurities.

[0032] EVs were enriched by ultracentrifugation (100,000-200,000 g, 1-2 hours) or size exclusion chromatography.

[0033] Purification was further performed using sucrose density gradient centrifugation or tangential flow filtration techniques to obtain high-purity white peony extracellular vesicles.

[0034] Formulation involves mixing purified EVs with pharmaceutical excipients, then lyophilizing, dispensing, or formulating them into the desired dosage form.

[0035] Example 3: As Figure 5 and Figure 6 The application of white peony extracellular vesicle nanoparticles in the preparation of drugs for the prevention or treatment of inflammatory diseases, namely inflammatory bowel disease.

[0036] An acute colitis model was established in C57BL / 6 mice by drinking 3% DSS solution for 5 days. Starting from day 1, mice were administered 50 mg / kg of Paeonia lactiflora EVs by gavage daily. PBS was used as a negative control and mesalazine as a positive control. Body weight, stool characteristics, and occult blood were recorded daily, and the DAI score was calculated. On day 7, the mice were sacrificed, colon length was measured, and colon tissue was collected for H&E staining and CD4 immunohistochemical analysis. The results showed that the DAI score of the Paeonia lactiflora EVs treatment group was significantly lower than that of the PBS group, colon length shortening was significantly improved, and colonic inflammatory infiltration and damage were reduced.

[0037]

[0038] Table 2. Experimental data on the treatment of colitis model

[0039] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A peony extracellular vesicle nanoformation for the treatment of inflammatory diseases, characterized in that, Its active ingredient is extracellular vesicles extracted from white peony tissue.

2. The peony extracellular vesicle nanoformation for the treatment of inflammatory diseases according to claim 1, characterized in that, The extracellular vesicles have a particle size of 50-200 nm.

3. The peony extracellular vesicle nanoformation for the treatment of inflammatory diseases according to claim 1, characterized in that, The extracellular vesicles can promote the differentiation of regulatory T cells and inhibit the production of inflammatory cytokines IFN-γ, IL-17A and TNF-α.

4. A method for preparing the Paeonia lactiflora extracellular vesicle nanoformation as described in any one of claims 1-3, characterized in that, Includes the following steps: White peony tissue homogenate, differential centrifugation to remove impurities, ultracentrifugation or chromatography to enrich and purify extracellular vesicles.

5. The use of the white peony extracellular vesicle nanoformation as described in any one of claims 1-3 in the preparation of a medicament for the prevention or treatment of inflammatory diseases.

6. The application according to claim 5, characterized in that, The inflammatory disease mentioned is inflammatory bowel disease.

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