Application of salvianolic acid B in the preparation of drugs for treating IPEX

Abstract

The present invention relates to the field of medical technology, and specifically to the use of salvianolic acid B in the preparation of drugs for treating IPEX. The present invention proves through in vivo animal experiments that salvianolic acid B has a good anti-inflammatory effect and can effectively alleviate the autoimmune reaction of Treg-deficient SF mice. In the present invention, the cations in the salvianolic acid B salts can be iron ions, calcium ions, sodium ions, potassium ions or magnesium ions, preferably magnesium ions; the dosage form of the drug for treating IPEX can be any dosage form described in pharmacy, and can be an oral liquid form, an injection form, a tablet form or a capsule form. By utilizing the key role of salvianolic acid B in IPEX disease, it can be prepared into a drug, providing a theoretical basis and a new treatment idea for the treatment of primary immunodeficiency diseases with traditional Chinese medicine; salvianolic acid B is easy to obtain, can be quickly put into production and obtain a high-efficiency preparation, and has important application value.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to application of salvianolic acid B in preparing a medicine for treating IPEX. Background Art

[0002] IPEX (immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome) is a rare, X-linked recessive immunodeficiency disorder. IPEX is characterized by multi-organ autoimmunity, manifesting as multiple endocrine autoimmune disorders, including rash, intractable diarrhea, type 1 diabetes, developmental failure, and autoimmune thyroiditis. Without prompt treatment, children with IPEX often die within one to two years of age.

[0003] IPEX is caused by mutations in the FOXP3 gene, which leads to developmental and differentiation disorders and dysfunction of regulatory T cells (Treg). Treg cells are characterized by the continuous expression of CD25 and FOP3 transcription factors, accounting for CD4 + T cells, which make up 5%-10% of the immune system, play a crucial role in maintaining immune homeostasis and mediating autoimmune tolerance. Treg-deficient SF mice can exhibit a disease process similar to IPEX. Male mice carrying the FOXP3 gene mutation exhibit Treg function deficiency, growth retardation, a scaly rash on the skin, lymphoproliferation (including enlarged liver, spleen, and lymph nodes), elevated plasma inflammatory cytokines, and a series of autoimmune inflammatory symptoms. They die within one month of birth, making them an ideal animal model for studying IPEX.

[0004] To date, immunosuppressive drugs combined with total parenteral nutrition and intravenous blood transfusions may alleviate clinical symptoms in children with IPEX. Hematopoietic stem cell transplantation is considered the only effective treatment for IPEX, but finding a suitable donor is often time-consuming and often leads to transplant-related chronic diseases. Therefore, before finding a suitable donor for transplantation, initial immunosuppressive therapy targeting T cells can alleviate clinical symptoms and increase the chance of survival.

[0005] Salvia miltiorrhiza (Danshen) is a traditional Chinese medicinal material, recorded in the Shennong's Herbal Classic. It has a bitter taste and slightly cold properties, and enters the heart and liver meridians. It has the effects of promoting blood circulation, removing blood stasis, relieving pain, cooling blood, and eliminating carbuncles. Salvianolic acid B (Sal-B) is the most pharmacologically active component of the water-soluble substances in Danshen. As an oligomeric compound, it contains several phenolic hydroxyl groups and therefore has strong antioxidant properties, can scavenge oxygen free radicals, and inhibit lipid peroxidation. It is widely used in the preparation of drugs for the prevention and treatment of cardiovascular diseases. In the prior art, most pharmaceutical companies use danshensu and protocatechuic aldehyde as quality control indicators for Danshen injections. Although Salvianolic acid B salts are also used in drugs for the prevention of cardiovascular and cerebrovascular diseases, the prevention of Alzheimer's disease, the treatment of renal failure, the improvement of uremic symptoms, antioxidant, anti-thrombotic, anti-cardiocerebral ischemia, liver protection, and blood lipid regulation, its use in the preparation of drugs for the treatment of IPEX has not been seen. Summary of the Invention

[0006] In view of the shortcomings of the prior art, the present invention aims to provide an application of salvianolic acid B in the preparation of a drug for treating IPEX.

[0007] To achieve the above object, the technical solution adopted by the present invention is as follows:

[0008] Application of salvianolic acid B in the preparation of drugs for treating IPEX.

[0009] Furthermore, in the salt of salvianolic acid B, the cation is iron ion, sodium ion, potassium ion or magnesium ion, preferably magnesium ion.

[0010] Furthermore, the dosage form of the drug for treating IPEX can be any dosage form described in pharmacy, such as oral liquid form, injection form, tablet form or capsule form.

[0011] Furthermore, the drug for treating IPEX can be used to treat one or more of the following IPEX symptoms: growth retardation, rash, intractable diarrhea, type 1 diabetes, stunted growth, autoimmune thyroiditis, enlargement of the liver, spleen, and lymph nodes, and elevated plasma inflammatory cytokines. Beneficial effects

[0012] The present invention demonstrates through in vivo animal experiments that, after Treg-deficient SF mice are treated with salvianolic acid B, their body size, spleen and peripheral lymph nodes are significantly reduced compared with SF control mice, the spleen index is reduced by 40.90%, and the lymphocyte infiltration areas of the liver and lung are reduced by 69.15% and 92.66%, respectively. In addition, the expression levels of IFN-r, IL-4 and IL-6 cytokines in Treg-deficient SF mice treated with salvianolic acid B are reduced by 64.81%, 77.97% and 56.47%, respectively, compared with SF control mice.

[0013] 2. The key role of salvianolic acid B in the present invention in IPEX disease can be utilized to prepare it into a medicine for the treatment of IPEX disease, providing a theoretical basis and new treatment ideas for the treatment of primary immunodeficiency diseases with traditional Chinese medicine. In addition, salvianolic acid B is easy to obtain, can be quickly put into production and obtain a highly effective preparation, and has important application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 It is a statistical graph of mouse survival time;

[0015] Figure 2 It is a phenotypic diagram of mouse body size, spleen, and peripheral lymph nodes;

[0016] Figure 3 This is a statistical graph of mouse spleen index data;

[0017] Figure 4 This is a statistical diagram of HE staining and infiltration area of ​​mouse liver and lung lymphocytes; Figure 4 A is the HE staining of mouse liver and lung lymphocytes. Figure 4 B is the statistical diagram of the lymphocyte infiltration area of ​​mouse liver. Figure 4 C is a statistical graph of the lymphocyte infiltration area in mouse lungs;

[0018] Figure 5 It is a statistical chart of the expression levels of mouse IFN-r, IL-4 and IL-6 cytokines; Figure 5 A is a statistical graph of mouse IFN-r cytokine expression levels, Figure 5 B is the statistical graph of mouse IL-4 cytokine expression levels, Figure 5 C is a statistical graph of mouse IL-6 cytokine expression levels. DETAILED DESCRIPTION

[0019] The technical solutions of the present invention are further described below with reference to the following examples, but the scope of protection of the present invention is not limited thereto. Unless otherwise defined, the technical terms used in the following examples have the same meanings as commonly understood by those skilled in the art to which the present invention belongs. The experimental reagents used in the following examples, unless otherwise specified, are all conventional biochemical reagents; the experimental methods described, unless otherwise specified, are all conventional methods.

[0020] In the present invention, the salvianolic acid B can be isolated and extracted from plants by various methods known in the art or modified methods, synthesized or prepared by semi-synthetic methods, or can be directly purchased commercially. The salvianolic acid B used in the present invention was purchased from Shanghai Ronghe Pharmaceutical Technology Development Co., Ltd.

[0021] The Treg-deficient SF mice involved in the examples all met the diagnostic criteria, and all indicators of the healthy control group were normal.

[0022] Normal male C57BL / 6J mice used in the examples were purchased from Beijing Huafukang Biotechnology Co., Ltd., and heterozygous female B6.Cg-Foxp3sf / J mice carrying a FOXP3 gene mutation were purchased from the Jackson Laboratory in the United States. Among the offspring mice bred after this, male mice carrying the FOXP3 gene mutation developed Treg function deficiency, which became the SF mice used in the present invention.

[0023] Example 1: Effects of salvianolic acid B on various indicators of Treg-deficient SF mice

[0024] Starting from the 8th day after birth, SF mice were randomly divided into 4 groups (6 mice in each group), and salvianolic acid B was dissolved in dH2O to prepare three different concentrations of salvianolic acid B solutions: 100 mg / kg, 300 mg / kg, and 500 mg / kg. Each group of mice was given dH2O and different concentrations of drugs for gavage treatment once a day for two weeks. Through weight monitoring, clinical scores and survival time observation, 500 mg / kg was screened as the therapeutic drug concentration for subsequent experiments. The survival time of mice after salvianolic acid B treatment was statistically analyzed, and a survival curve was drawn. The survival time of mice in each experimental group is shown in Figure 2. Figure 1 As shown, the results showed that compared with SF control mice, the survival time of the drug intervention group was significantly prolonged, ranging from 42 days to 80 days. There was no decrease in survival rate in the normal mice treated with salvianolic acid B and the blank control group, indicating that salvianolic acid B can prolong the survival time of Treg-deficient SF mice and will not affect the survival time of normal mice.

[0025] The experiment was conducted on normal mice and SF mice from the same batch. The normal mice were randomly divided into two groups, one group was blank control, and the other group was normal mice treated with salvianolic acid B. The SF mice were randomly divided into two groups, one group was SF control group, and the other group was SF mice treated with salvianolic acid B (6 mice in each group). The normal mice treated with salvianolic acid B and the SF mice treated with salvianolic acid B were treated by gavage with 500 mg / kg salvianolic acid B solution, while the blank control and SF control groups were treated by gavage with dH2O equal to the amount of salvianolic acid B. After the mice were treated with gavage once a day for two weeks, various indicators were measured. The mice in each experimental group were anesthetized with chloral hydrate and then killed. The spleen, liver, lung, small intestine and other tissues were taken for comparative observation. The results are as follows: Figure 2The results showed that compared with blank control mice, normal mice treated with salvianolic acid B lost weight, had delayed development, and had obvious dermatitis on the ears and tails. In addition, the spleen and peripheral lymph nodes were significantly enlarged. However, after Treg-deficient SF mice were treated with salvianolic acid B, their body size, spleen and peripheral lymph nodes were significantly reduced compared with SF control mice. The body weight and spleen weight of mice were weighed, and the spleen index of each group of mice was calculated, as shown in Figure 5. Figure 3 As shown in Figure 2, compared with normal mice, the spleen index of SF mice increased significantly. Compared with SF control mice, the spleen index of SF mice after drug treatment decreased by 40.90%. In order to compare the inflammatory cell infiltration of animals in different groups, the spleens and livers of mice in each experimental group were embedded in paraffin and then sectioned, and then HE staining was performed. The staining results and data statistics are shown in Figure 2. Figure 4 , Figure 4 A shows that compared with the blank control mice, the liver and lungs of the normal mice treated with salvianolic acid B showed obvious lymphocyte infiltration. Compared with the SF control mice, the lymphocyte infiltration area of ​​the liver and lungs of the SF mice after drug treatment was significantly reduced. The statistical data showed that the reduction was 69.15% ( Figure 4 B), 92.66% ( Figure 4 C); The serum cytokine levels of mice in each experimental group were measured by enzyme-linked immunosorbent assay (ELISA). The results were as follows Figure 5 As shown in the figure, compared with the blank control mice, the expression levels of IFN-r, IL-4, and IL-6 cytokines in the normal mice treated with salvianolic acid B were significantly increased, and the expression levels of IFN-r ( Figure 5 A), IL-4 ( Figure 5 B), IL-6 ( Figure 5 C) Cytokine expression levels were reduced by 64.81%, 77.97%, and 56.47% compared with those in SF control mice. Figure 2-Figure 5 As shown, there was no significant difference in various indicators between the normal mice treated with salvianolic acid B and the blank control group, indicating that salvianolic acid B has no toxic side effects.

[0026] Our laboratory conducted multiple experiments using the procedures of Example 1, demonstrating that the expression levels of IFN-r, IL-4, and IL-6 cytokines in Treg-deficient SF mice treated with salvianolic acid B were significantly reduced compared to those in SF control mice. Neither the salvianolic acid B-treated nor the blank control groups showed a decrease in survival rate in normal mice, indicating that salvianolic acid B can prolong the survival of Treg-deficient SF mice without affecting the survival of normal mice. These results demonstrate that salvianolic acid B has a potent anti-inflammatory effect and can effectively alleviate autoimmune responses in Treg-deficient SF mice.

Claims

1. Application of salvianolic acid B in the preparation of drugs for the treatment of X-linked polyendocrine enteropathy with immune dysregulation syndrome.

2. The use according to claim 1, characterized in that The salvianolic acid B also includes salvianolic acid B salt, wherein the cation of the salvianolic acid B salt is iron ion, calcium ion, sodium ion, potassium ion or magnesium ion.

3. The use according to claim 2, characterized in that The cation of the salvianolic acid B salt is a magnesium ion.

4. The use according to any one of claims 1 to 3, characterized in that The dosage form of the drug is any dosage form described in pharmacy.

5. The use according to claim 4, characterized in that The dosage form of the medicine is oral liquid, injection, tablet or capsule.

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