Use of an aDRB2 agonist in the manufacture of a medicament for the prevention and / or treatment of sepsis

Abstract

This invention relates to the field of pharmaceuticals, specifically to the application of an ADRB2 agonist in the preparation of drugs for the prevention and / or treatment of sepsis. Cellular and animal experiments have shown that knocking out Adrb2 in B cells leads to impaired IL-10 expression and increased mortality in sepsis model mice, while administration of an ADRB2 activator reduces mortality in these mice. This suggests that ADRB2 agonists can be used for the prevention and / or treatment of sepsis, providing a new approach for the clinical application of sepsis medication.

Description

[0001] This application is a divisional application of patent application number 2025100097429, filed on January 3, 2025. Technical Field

[0002] This invention relates to the field of pharmaceuticals, and more specifically, to the use of an ADRB2 agonist in the preparation of medicaments for the prevention and / or treatment of sepsis. Background Technology

[0003] Sepsis occurs when the immune system is overactivated in cases of severe infection of organs (such as the lungs and digestive tract) or when pathogens (such as LPS) enter the bloodstream in large quantities. During these pathological processes, the excessive release of inflammatory factors such as TNFα, IL-6, and IFN-γ, or inflammatory substances such as reactive oxygen species, by the immune response leads to multi-organ damage, disseminated intravascular coagulation, and even death. Currently, treatment options for sepsis are extremely limited, and it is virtually impossible to predict or prevent this fatal disease in clinical practice.

[0004] ADRB2 receptors are widely expressed in immune cells, and studies have suggested their potential association with various human diseases. Currently, β-receptor agonists, as a successful clinical application, have been used to improve airway obstruction caused by asthma, chronic obstructive pulmonary disease, and pulmonary fibrosis-related lung dysfunction. However, despite the significant attention given to the potential therapeutic value of ADRB2 in various diseases, there are currently no reports on its application in the treatment of sepsis. This indicates that the specific mechanism of action and therapeutic potential of ADRB2 in sepsis requires further investigation to provide possible directions for developing novel treatment strategies. Summary of the Invention

[0005] To fill the gap in the prior art, the present invention provides the use of an ADRB2 agonist in the preparation of a medicament for the prevention and / or treatment of sepsis.

[0006] In one implementation, the application includes enhancing IL-10 expression in regulatory B cells by using an ADRB2 agonist.

[0007] In one implementation, the application includes reducing a patient's susceptibility to sepsis by using an ADRB2 agonist.

[0008] In one embodiment, the application includes directly promoting the anti-inflammatory function of IL-10-positive regulatory B cells via the ADRB2 receptor.

[0009] In a preferred embodiment, the ADRB2 agonist is formoterol. Attached Figure Description

[0010] 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: Figure 1 Cd19-Cre;Adrb2 fl / fl and littermate Adrb2 fl / fl control mice were subjected to LPS-induced sepsis. The IL-10 mRNA levels in the spleen, liver and inguinal lymph nodes of mice were detected by qPCR 1 hour after LPS treatment. Figure 2 Cd19-Cre;Adrb2 fl / fl and littermate Adrb2 fl / fl control mice were subjected to LPS-induced sepsis. The plasma IL-10 protein level of the mice was detected by ELISA 1 hour after LPS treatment. Figure 3 The survival rate of Cd19-Cre;Adrb2 fl / fl and littermate Adrb2 fl / fl control mice was measured daily after LPS-induced sepsis. Figure 4 CD45+CD19+CD24hiCD27+ B cells were isolated from human spleens and divided into three groups: saline treatment group (control group), LPS treatment group (B cells under simulated sepsis conditions), and LPS+formoterol treatment group (B cells under simulated sepsis conditions and treated with formoterol). The expression levels of IL-10 mRNA and protein were detected by qPCR and ELISA.

[0011] Figure 5 C57BL / 6 wild-type mice underwent splenectomy and sympathectomy. A sepsis model was induced by LPS. The mice were then treated with saline (control group) and formoterol (experimental group). The IL-10 mRNA level in the spleen of mice 1 hour after LPS treatment was detected by qPCR. The IL-10 protein level in the plasma of mice 1 hour after LPS treatment was detected by ELISA. Figure 6 C57BL / 6 wild-type mice underwent splenectomy and sympathectomy. A sepsis model was induced using LPS. The mice were then treated with saline (control group) and formoterol (experimental group), respectively. The survival rate of the mice was measured daily. Detailed Implementation

[0012] 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.

[0013] Example 1: Establishment of a mouse sepsis model Cd19-Cre and Adrb2 mice were obtained by crossing the B-cell-specific tool mouse Cd19-Cre mouse (Cyagen Biosciences, #C001204) with Adrb2 fl / fl mice (Cyagen Biosciences, #S-CKO-01084). fl / fl Mouse strains were used to achieve specific deletion of Adrb2 in B cells.

[0014] For the LPS-induced sepsis model, LPS (dissolved in sterile PBS at a concentration of 1 mg / mL) was intravenously injected at a dose of 10 mg / kg body weight into mice meeting specific conditions (including control wild-type mice (C57BL / 6J) and CD19-Cre;Adrb2). fl / fl B-cell conditional knockout mice).

[0015] For a cecal ligation and puncture (CLP)-induced sepsis model, mice under specified conditions were anesthetized with 3% isoflurane, and the abdominal skin was shaved and treated with iodine and alcohol. A longitudinal incision was made along the midline of the skin and peritoneum to expose the cecum. The cecum was ligated with 5 / 0 silk suture approximately 1 cm distal to the ligation. The distal end of the ligated cecum was punctured twice with a 22-gauge needle. The sham surgery consisted of all steps except for the unligated or unpunctured cecum. The incisions on the peritoneum and skin were then sutured.

[0016] After establishing the animal model, cytokine levels (such as IL-10, TNF-α, and IFN-β) in the spleen and plasma were detected by qPCR and ELISA. Significant changes in IL-10 levels were an important indicator of the success of the endotoxemia model. Furthermore, changes in the spleen's immune microenvironment were observed using immunolabeling techniques (such as TH-labeled sympathetic nerves and CD169-labeled marginal zone macrophages). More importantly, the survival rate of the experimental animals was recorded to assess the severity of sepsis and the success of the model. A decreased survival rate indicated the establishment of sepsis.

[0017] Example 2: Specific knockout of Adrb2 receptors in B cells of a sepsis model mouse led to increased mortality. Compared with LPS-induced control mice (C57BL / 6J), Cd19-Cre;Adrb2 fl / fl In mice, IL-10 expression in the spleen was significantly inhibited after LPS treatment, while IL-10 expression levels in the liver or inguinal lymph nodes appeared to be unaffected. Figure 1 This defective splenic IL-10 response also leads to a decrease in plasma IL-10 levels. Figure 2 This resulted in a decrease in the survival rate of Cd19-Cre;Adrb2 fl / fl mice. Figure 3).

[0018] The above results indicate that knocking out the Adrb2 receptor on B cells in sepsis model mice leads to an increased mortality rate.

[0019] Example 3: The Adrb2 agonist formoterol promotes the expression of IL-10 in human B cells. CD45 was isolated from human spleen using flow cytometry. + CD19 + CD24 hi CD27 + B cells were divided into three groups: a saline treatment group (control group), an LPS treatment group (B cells under simulated sepsis conditions), and an LPS + formoterol treatment group (B cells under simulated sepsis conditions treated with formoterol). The expression levels of IL-10 mRNA and protein were detected by qPCR and ELISA. It was found that the ADRB2 agonist formoterol effectively enhanced the expression of IL-10 mRNA and protein production in LPS-induced human CD24hi CD27+ B cells. Figure 4 This indicates that formoterol directly promotes the anti-inflammatory function of IL-10-positive regulatory B cells through the ADRB2 receptor.

[0020] Example 4: Adrb2 agonist formoterol inhibits mortality in septic mice Immediately after LPS-induced establishment of a sepsis mouse model, formoterol (Selleckchem, dissolved in 0.5 mg / ml sterile PBS) was administered intravenously at a dose of 5 mg / kg body weight. Compared with control mice (LPS-induced C57BL / 6J wild-type mice), this Adrb2 agonist promoted the production and secretion of IL-10 in mice. Figure 5 This significantly suppressed the mortality rate in septic mice, thereby allowing for the complete restoration of disease susceptibility in these mice. Figure 6 ).

[0021] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A method for establishing a mouse sepsis model, characterized in that: The method includes: By comparing B-cell-specific tool mice, Cd19-Cre mice, with Adrb2 fl / fl Mouse hybridization yielded Cd19-Cre;Adrb2 fl / fl Mouse strains were used to achieve specific deletion of Adrb2 in B cells; LPS at a concentration of 1 mg / mL dissolved in sterile PBS was intravenously injected into Cd19-Cre at a dose of 10 mg / kg body weight; Adrb2 fl / fl B-cell conditional knockout mice were used to establish a mouse sepsis model. After obtaining the sepsis mouse model through LPS induction, the ADRB2 agonist formoterol was dissolved in 0.5 mg / mL sterile PBS and immediately injected intravenously at a dose of 5 mg / kg body weight to promote the production and secretion of IL-10 in mice, which significantly inhibited the mortality rate of sepsis mice.

2. The method according to claim 1, characterized in that: Cd19-Cre; Adrb2 fl / fl The expression of IL-10 in the spleen of mice was significantly inhibited after LPS treatment.

3. The method according to claim 2, characterized in that: Cd19-Cre; Adrb2 fl / fl Mice also showed decreased plasma IL-10 levels after LPS treatment, resulting in Cd19-Cre and Adrb2. fl / fl The survival rate of mice decreased.

4. The method according to claim 1, characterized in that: Knockout of the Adrb2 receptor in B cells of sepsis model mice led to an increased mortality rate in the mice.

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