Application of STING agonist MSA-2 manganese salt in preparation of product for activating immune effect

Abstract

The invention provides application of STING agonist MSA-2 manganese salt in preparation of a product for activating an immune effect, and relates to the technical field of biomedicine. The invention provides an STING agonist MSA-2 manganese salt and regulation and control of immune response generated by combination of the STING agonist MSA-2 manganese salt and radiotherapy. According to the method provided by the invention, the STING agonist MSA-2 is modified by using metal cations Mn, and the immune response is efficiently activated by exogenously adding Mn, so that the generation of anti-tumor effect related factors is remarkably increased. The STING agonist drug MSA-2-Mn containing the cation Mn provided by the invention not only stimulates the regulation and control of the spleen on innate immunity and adaptive immunity, but also can significantly enhance the immune activation effect generated by radiotherapy and reduce toxic and side effects, and develops a good prospect for a tumor treatment method.

Description

Technical Field

[0001] The present application belongs to the field of biomedical technology, and specifically relates to the application of STING agonist MSA-2 manganese salt in the preparation of products for activating immune effects. Background Art

[0002] Immunotherapy has good anti-tumor effects and has occupied an important position in the clinical treatment of tumors. Studies have confirmed that the non-nucleotide STING agonist MSA-2, due to its special non-covalent dimer structure and acidic properties, can efficiently activate the STING signaling pathway, significantly induce the increase of type I interferon (IFN-β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in tumors and plasma, thereby playing an anti-tumor immune effect. The STING agonist MSA-2 has been used in extensive preclinical studies, and its application prospects are promising.

[0003] Manganese ion (Mn 2+ ) is highly sensitive to the cGAS-STING pathway and can directly activate cGAS, enhancing the immune response by promoting antigen uptake and antigen presentation. 2+ Combination therapy with STING agonists showed more effective anti-tumor immune responses. Therefore, we synthesized the manganese salt of the STING agonist MSA-2 (MSA-2-Mn) and studied its immune activation effects in animals.

[0004] At the same time, radiotherapy has always been the main treatment for many solid tumors, and radiotherapy combined with immunotherapy has a good therapeutic effect. Therefore, we also studied the immune activation effect of MSA-2-Mn on animals after radiotherapy. Summary of the invention

[0005] The present application provides the use of STING agonist MSA-2 manganese salt in the preparation of products that activate immune effects, so as to prepare products that activate immune effects.

[0006] In order to solve the above technical problems, a technical solution adopted in this application is:

[0007] Use of STING agonists in the preparation of products for activating immune effects.

[0008] Preferably, the STING agonist is a manganese salt of MSA-2, wherein the manganese salt of the STING agonist MSA-2 is MSA-2-Mn.

[0009] Preferably, the chemical formula of the MSA-2-Mn is as follows:

[0010]

[0011] Preferably, the MSA-2-Mn has the function of activating the cGAS-STIG signaling pathway and increasing the secretion of IL-6, IFN-γ and TNF-α.

[0012] Preferably, the MSA-2-Mn has the function of activating the cGAS-STING signaling pathway and increasing the secretion of IL-6, IFN-γ and TNF-α after combined radiotherapy.

[0013] Preferably, the STING agonist MSA-2-Mn is included.

[0014] Preferably, the product is an injection or an infusion.

[0015] The MSA-2-Mn in this application can efficiently activate the cGAS-STING signaling pathway, significantly increase the secretion of IL-6, IFN-γ and TNF-α, and enhance the spleen's regulation of innate immunity and adaptive immunity. At the same time, this application can achieve that MSA-2-Mn can still efficiently activate the cGAS-STING signaling pathway after radiotherapy, significantly increase the secretion of IL-6, IFN-γ and TNF-α, and enhance the spleen's regulation of innate immunity and adaptive immunity.

[0016] In summary, the STING agonist drug MSA-2-Mn containing cationic Mn provided in the present application not only stimulates the spleen's regulation of innate immunity and adaptive immunity, but also can significantly enhance the immune activation effect produced by radiotherapy and reduce toxic side effects, opening up good prospects for tumor treatment methods. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is a diagram of the process of injection experiments in male mice using different methods;

[0018] Figure 2 This is a graph showing the expression level of IL-6 in mouse serum detected by ELISA;

[0019] Figure 3 This is a graph showing the expression level of IFN-γ in mouse serum detected by ELISA;

[0020] Figure 4 This is a graph showing the expression level of TNF-α in mouse serum detected by ELISA;

[0021] Figure 5 This is the histomorphological staining (HE staining) of the spleen after treatment with MSA-2 and MSA-2-Mn;

[0022] Figure 6 The spleen index of mice treated with MSA-2 and MSA-2-Mn. DETAILED DESCRIPTION

[0023] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention is further described in detail below in conjunction with specific embodiments.

[0024] In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention may also be implemented in other ways than those described herein. Therefore, the present invention is not limited to the specific embodiments of the following disclosure.

[0025] The use of a STING agonist in the preparation of a product for activating immune effects, wherein the STING agonist is a manganese salt of MSA-2, wherein the manganese salt of the STING agonist MSA-2 is MSA-2-Mn. The chemical formula of the MSA-2-Mn is as follows:

[0026]

[0027] Preferably, the synthesis path of the MSA-2-Mn is as follows:

[0028]

[0029] Preferably, the product comprises the STING agonist MSA-2-Mn; the product is an injection or an infusion. The preparation method of the MSA-2-Mn is as follows:

[0030] S1: Weigh 58.8 mg (0.2 mmol) of 4-(5,6-dimethoxybenzo[B]thiophen-2-yl)-4-oxobutanoic acid and 8.0 mg (0.2 mmol) of sodium hydroxide into a reactor, add purified water, react at room temperature for 12 hours, stop the reaction, and keep it for the next step without further treatment;

[0031] S2: Weigh 12.6 mg (0.1 mmol) of manganese chloride, dissolve it in purified water and add it to the solution of the first step. React at room temperature for 24 hours. Stop the reaction after the reaction is complete. Filter out the solid, recrystallize it with water and dry it to obtain 52.56 mg of the compound, recorded as MSA-2-Mn.

[0032] Preferably, MSA-2-Mn efficiently activates the cGAS-STING signaling pathway, significantly increases the secretion of IL-6, IFN-γ and TNF-α, and enhances the spleen's regulation of innate and adaptive immunity. It is achieved that after radiotherapy, MSA-2-Mn can still efficiently activate the cGAS-STING signaling pathway, significantly increase the secretion of IL-6, IFN-γ and TNF-α, and enhance the spleen's regulation of innate and adaptive immunity.

[0033] Embodiment 1:

[0034] The immune effects of STING agonists MSA-2 and MSA-2-Mn are activated, and their immune effects after radiotherapy are evaluated.

[0035] Nine-week-old C57BL / 6 male mice were selected and divided into two groups: Treatment 1 group (control group) and Treatment 2 group (radiotherapy group). The mice in Treatment 1 group were divided into three subgroups: Control, MSA-2, MSA-2-Mn (n = 5 per group), and the mice in Treatment 2 group were divided into three subgroups: IR, IR-MSA-2, IR-MSA-2-Mn (n = 5 per group).

[0036] On day 0, the Treatment 1 group received no treatment, while the Treatment 2 group received 4 Gy X-ray irradiation radiotherapy. Subsequently, on days 1, 4, 7, 10, and 13, the mice were subcutaneously injected with a 50 mg / kg dose of vehicle, MSA-2, and MSA-2-Mn, respectively.

[0037] See also Figure 1 , Figure 1 This is a process diagram of injection experiments in male mice by different methods. The mice were cultured in the same environment and killed on the 14th day. Treatment method 1 for male C57BL / 6 mice: The mice were divided into three groups: Control, MSA-2 (50 mg / kg) and MSA-2-Mn (50 mg / kg). Control, 50 mg / kg MSA-2, 50 mg / kg MSA-2-Mn were injected subcutaneously once every 3 days for 5 consecutive injections, and the mice were killed 1 day later. Treatment method 2 for male C57BL / 6 mice: The mice were divided into three groups: IR+Control, IR+MSA-2 (50 mg / kg) and IR+MSA-2-Mn (50 mg / kg). First, a dose of 4Gy of X-rays was used for irradiation. Then, Control, 50 mg / kg MSA-2, 50 mg / kg MSA-2-Mn were injected subcutaneously once every 3 days for 7 consecutive injections, and the mice were killed 1 day later.

[0038] See also Figure 2-4 , Figure 2-4The expression levels of IL-6, IFN-γ and TNF-α in mouse serum were detected by ELISA. The levels of TNF-α, IFN-γ and IL-6 in the serum of each group were determined by enzyme-linked immunosorbent assay (ELISA). The experimental results showed that compared with the control group, the levels of TNF-α, IFN-γ and IL-6 in the serum of the experimental groups treated with MSA-2 and MSA-2-Mn were significantly increased. In addition, whether in the case of single treatment or combined radiotherapy, the levels of IFN-γ and TNF-α in the serum of the MSA-2-Mn-treated group increased more significantly than those of the experimental group treated with MSA-2. At the same time, this also confirms that the use of MSA-2 and MSA-2-Mn can effectively activate the immune system, and after combined radiotherapy, MSA-2 and MSA-2-Mn still effectively activate the immune system.

[0039] See also Figure 5-6 , Figure 5 This is the histomorphological staining (HE staining) of the spleen after treatment with MSA-2 and MSA-2-Mn; Figure 6 The spleen index of mice treated with MSA-2 and MSA-2-Mn was shown in Figure 2. The spleen tissue of mice was weighed and histologically analyzed. The spleen index (i.e., the ratio of spleen weight to body weight) after treatment with MSA-2 and MSA-2-Mn increased significantly, whether in the case of single treatment or combined treatment after radiotherapy. Under the light microscope, the red pulp area of ​​the spleen was significantly enlarged, accompanied by lymph follicle hyperplasia, splenic congestion and sinusoid dilatation, and splenic corpuscle hyperplasia.

[0040] In summary, the STING agonist MSA-2 manganese salt provided in this application and the immune activation effect produced by its combined treatment with radiotherapy promote the body's secretion of TNF-α, IFN-γ and IL-6 to activate immunity. The STING agonist drug MSA-2-Mn containing cationic Mn provided in this application not only stimulates the spleen's regulation of innate immunity and adaptive immunity, but also can significantly enhance the immune activation effect produced by radiotherapy and reduce toxic side effects, opening up good prospects for tumor treatment methods.

[0041] The above description is only an embodiment of the present application and does not limit the patent scope of the present application. Any equivalent structure or equivalent process transformation made using the contents of the present application specification and drawings, or directly or indirectly used in other related technical fields, are also included in the patent protection scope of the present application.

Claims

1. Use of a STING agonist in the preparation of a product for activating immune effects, characterized in that: The STING agonist is a manganese salt of MSA-2, wherein the manganese salt of the STING agonist MSA-2 is MSA-2-Mn.

2. The use according to claim 1, characterized in that: The chemical formula of the MSA-2-Mn is as follows:

3. The use according to claim 1, characterized in that: The MSA-2-Mn has the function of activating the cGAS-STING signaling pathway and increasing the secretion of IL-6, IFN-γ and TNF-α.

4. The use according to claim 1, characterized in that: The MSA-2-Mn has the function of activating the cGAS-STING signaling pathway and increasing the secretion of IL-6, IFN-γ and TNF-α after combined radiotherapy.

5. A product for activating immune effects, characterized in that: Contains the STING agonist MSA-2-Mn.

6. A product for activating immune effects according to claim 5, characterized in that: The product is an injection or an infusion.

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