Preparation of copper salt of STING agonist MSA-2 and lipid nanomaterial combined with copper salt, and application of copper salt and lipid nanomaterial in tumor treatment
The combined application of STING mRNA delivery through the copper salt of MSA-2 and lipid nanomaterials to activate the cGAS-STING signaling pathway, solving the problem of limited effectiveness of existing MSA-2 in tumor immunotherapy, significantly reducing tumor growth and improving survival , providing strategies for enhancing tumor immunotherapy.
Patent Information
- Application Number
- CN202311729623.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-12-15
- Publication Date
- 2024-12-13
AI Technical Summary
The existing STING agonist MSA-2 has limited effect in tumor immunotherapy, and it is difficult to effectively activate the cGAS-STING signaling pathway, and its stability and targeting ability are insufficient.
The STING mRNA (1AP27@STING mRNA) delivered by MSA-2 copper salt (MSA-2-Cu) combined with lipid nanomaterials was used to activate the cGAS-STING signaling pathway through Cu, and the expression of STING protein was improved by phospholipid nanoparticles. Enhance tumor immune response.
It significantly reduces tumor growth, improves the survival rate of mice, improves the anti-tumor effect of MSA-2, activates a strong immune response, and provides a strategy to enhance tumor immunotherapy.
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Figure CN119112943A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of biomedical technology, and in particular to the preparation of a copper salt of a STING agonist MSA-2 and its application in tumor treatment drugs, as well as the preparation of a lipid nanomaterial 1AP27@STING mRNA and its application in further enhancing tumor immunotherapy. Background Art
[0002] Cancer immunotherapy has revolutionized cancer treatment strategies, from innate immunity to adaptive immunity through immune checkpoint blockade therapy, adoptive cell therapy, and vaccines. Activating cGAS / STING-dependent signal transduction to initiate innate immunity is becoming a promising anti-tumor strategy in clinical applications. In recent years, several STING agonists have shown good tumor immunotherapy effects, including small molecule STING agonists, cyclic dinucleotide (CDN) STING agonists, and metal ions (Zn 2+ and Mn 2+ ) and other STING agonists.
[0003] In recent years, the small molecule MSA-2 (benzothiopheneoxybutyric acid) has been developed as a non-nucleotide STING agonist with good antitumor activity. It is an orally available human STING agonist that can be used for systemic administration in clinical practice. A modified MSA-2 with selenium replacing sulfur has shown to be a viable antitumor drug. Nanoliposome vesicles encapsulating MSA-2 have improved stability and antitumor effects. Therefore, strategies to enhance the immunotherapy effect of MS A-2 are valuable for clinical applications. Summary of the invention
[0004] The purpose of the present invention is to enhance the immunotherapy effect of MSA-2.
[0005] In order to achieve the above object, the present invention adopts the following technical solutions:
[0006] The application of MSA-2-Cu in the preparation of tumor treatment products is characterized in that the MSA-2-Cu is the copper salt of MSA-2.
[0007] Preferably, the MSA-2-Cu releases Cu and MSA-2 at the tumor site, and Cu efficiently activates the cGAS-STING signaling pathway, while MSA-2 can selectively target tumor tissues and exhibit a higher ability to activate the STING pathway.
[0008] Preferably, the chemical formula of the MSA-2-Cu is as follows:
[0009]
[0010] Preferably, the synthesis path of the MSA-2-Cu is as follows:
[0011]
[0012] Preferably, the specific preparation method of the MSA-2-Cu is as follows:
[0013] S1: 4-(5,6-dimethoxybenzo[B]thiophen-2-yl)-4-oxobutanoic acid and sodium hydroxide were placed in a reactor, purified water was added, and the reaction was carried out at room temperature for 12 hours. The reaction was stopped and the reaction was not further processed and was reserved for the next step;
[0014] S2: Take copper sulfate, dissolve it in purified water and add it to the solution in the first step. Let it 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 a compound, recorded as MSA-2-Cu.
[0015] Preferably, the product further comprises STING mRNA delivered by lipid nanomaterials, and the lipid nanomaterials are phospholipid nanoparticles.
[0016] Preferably, the phospholipid nanocompound is 1AP27, and the STING mRNA delivered by the lipid nanomaterial is 1AP27@STING mRNA.
[0017] Preferably, the preparation method of the phospholipid nanocompound is as follows:
[0018] S1. 2-Chloro-1,3,2-dioxaphospholane-2-oxide was slowly added to a beaker containing alcohol and Et3N and stirred at -15°C for 5 minutes; then, the reaction was continued at 25°C for 12 hours; the precipitate was filtered and rinsed with ether three times. The mixture was filtered to remove triethylamine hydrochloride, and the filtrate was concentrated by rotary evaporation to obtain the alkylated dioxaphospholane molecule; the yield of all syntheses exceeded 90%;
[0019] S2. In a round-bottom flask, dissolve the alkylated dioxaphospholane molecules and amine into CH3CN;
[0020] The resulting mixture was heated to 70°C and stirred for 3d-5d; the reaction was monitored by TLC analysis and fractionated on a silica gel column with a solvent gradient of 4% methanol in dichloromethane to 20% methanol in dichloromethane; the final ionizable phospholipids were concentrated by rotary evaporation and dried under vacuum for 24h.
[0021] Preferably, lipid nanoparticles are synthesized for the delivery of STING mRNA according to the molar ratio of phospholipid nanocompound (1AP27): dioleoyl L-α-phosphatidylethanolamine (DOPE): cholesterol (Cholesterol): 1,2-dimyristoyl-RAC-glycero-3-methoxypolyethylene glycol 2000 (DMG-PEG2000) = 50:40:20:0.25.
[0022] In this application, two strategies for improving the anti-tumor therapy of MSA-2 are adopted. First, copper-modified MSA-2 is synthesized in this application, and the method shows that the cytotoxicity of Cu enhances the STING pathway. 81 ionizable phospholipid nanoparticles are also synthesized in this application, among which the phospholipid nanoparticles 1AP27@STING mRNA can effectively deliver STING-mRNA and increase the expression of STING protein. Combining 1AP27@STING mRNA with MSA-2-Cu can significantly reduce the growth of B16F10 tumors and increase the survival rate of mice. The 1AP27@STING mRNA+MSA-2-Cu treatment in this application stimulates a strong immune response by activating the cGAS-STING pathway, improves the anti-tumor effect of MSA-2, and provides a promising strategy for enhancing tumor immunotherapy. BRIEF DESCRIPTION OF THE DRAWINGS
[0023] Figure 1 A strategy for combining lipid nanomaterials to deliver STING mRNA (1AP27@STING mRNA) and MSA-2-Cu for tumor treatment.
[0024] Figure 2 The invention relates to the synthesis of ionizable phospholipid compounds; (A) Synthesis scheme of ionizable phospholipid compounds: ionizable phospholipids (nAPm) are obtained by coupling (B) amine (nA) with alkylated dioxaphospholane oxide molecules (Pm).
[0025] Figure 3 Optimization process of lipid nanomaterials; (A) Detection of cell viability after 24h treatment of MC38 cells with lipid nanomaterials delivering GFP plasmid at different concentrations (0.25, 0.5, 1.0 μg / mL). (B) Fluorescence images of MC38 cells treated with different lipid nanomaterials nAPm delivering GFP plasmid for 24h (three replicates per group). (C) Percentage of GFP-positive cells = (GFP-positive cells / total cells)%.
[0026] Figure 4To synthesize and test the in vitro toxicity of MSA-2-Cu; (A) Synthesis scheme of MSA-2-Cu. (B) Cell viability of MC38 cells after 24 h of treatment with MSA-2 and MSA-2-Cu. Mean±SD, (n=3). (C) Cell viability of B16F10 cells after treatment with MSA-2 and MSA-2-Cu. Mean±SD, (n=3). (D) Microscopic images of B16F10 cells after 24 h of treatment with 100μM MSA-2 and MSA-2-Cu (n=3).
[0027] Figure 5 The expression levels of p-TBK1, p-STING, and STING were detected by Western blot after RAW264.7 cells were treated with different concentrations of MSA-2 and MSA-2-Cu for 24 hours. β-actin was the internal reference protein.
[0028] Figure 6 1AP27@STING mRNA combined with MSA-2-Cu for tumor treatment: (A) Schematic diagram of the anti-tumor effect after intratumoral injection of Control (drug solvent), MSA-2, MSA-2-Cu, and 1AP27@STING mRNA + MSA-2-Cu: MSA-2, MSA-2-Cu group was injected with 150μg MSA-2, MSA-2-Cu, and 1AP27@STING mRNA + MSA-2-Cu group was first injected with 3μg 1AP27@STING mRNA, and 6 hours later, 150μg MSA-2-Cu was injected for a total of three times. (B) Individual mouse tumors in the Control, MSA-2, MSA-2-Cu, and 1AP27@STING + MSA-2-Cu groups (n = 10). (C) Mouse tumors in the Control, MSA-2, MSA-2-Cu, and 1AP27@STING mRNA + MSA-2-Cu groups (n = 10). (D) Changes in the survival rate of mice in the Control, MSA-2, MSA-2-Cu, and 1AP27@STINGmRNA+MSA-2-Cu groups (n=10 in each group). (*P<0.05; **P<0.01; ***P<0.001; ****P<0.001.).
[0029] Figure 7The anti-tumor effect of 1AP27@STING mRNA combined with MSA-2-Cu: (A) Schematic diagram of the mechanism of tumor treatment: Control, MSA-2, MSA-2-Cu and 1AP27@STING mRNA+MSA-2-Cu were intratumorally administered three times. The MSA-2 and MSA-2-Cu groups were given 150μg MSA-2 or MSA-2-Cu. The 1AP27@STING mRNA+MSA-2-Cu group was first given 3μg 1AP27@STING mRNA, and 150μg MSA-2-Cu was injected intratumorally 6h later. (B) Control, MSA-2, MSA-2-Cu, 1AP27@STING mRNA+MSA-2-Cu. The levels of IFN-β and Il-6 in mouse serum were detected by ELISA (n=5). (C) HE staining of tumor tissues in the Control, MSA-2, MSA-2-Cu, and 1AP27@STING+MSA-2-Cu groups after treatment (n=5). (D) Immunofluorescence images of CD4+ and CD8+ T cells in the Control, MSA-2, MSA-2-Cu, and 1AP27@STING+MSA-2-Cu groups after treatment (n=5). *p<0.05, **p<0.01, ***p<0.0001, ns indicates no statistical significance). DETAILED DESCRIPTION
[0030] The present invention is further described in detail below in conjunction with specific embodiments.
[0031] See also Figure 1 The present application provides the use of MSA-2-Cu in the preparation of a product for treating tumors, wherein the MSA-2-Cu is a copper salt of MSA-2, and the MSA-2-Cu releases Cu and MSA-2 at the tumor site, and efficiently activates the cGAS-STING signaling pathway through Cu, while MSA-2 can selectively target tumor tissues and show a higher ability to activate the STING pathway.
[0032] In one embodiment, the MSA-2-Cu chemical formula is as follows:
[0033]
[0034] The synthesis path of the MSA-2-Cu is as follows:
[0035]
[0036] The specific preparation method of the MSA-2-Cu is as follows:
[0037] S1: 4-(5,6-dimethoxybenzo[B]thiophen-2-yl)-4-oxobutanoic acid and sodium hydroxide were placed in a reactor, purified water was added, and the reaction was carried out at room temperature for 12 hours. The reaction was stopped and the reaction was not further processed and was reserved for the next step;
[0038] S2: Take copper sulfate, dissolve it in purified water and add it to the solution in the first step. Let it 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 a compound, recorded as MSA-2-Cu.
[0039] In one embodiment, the product further comprises STING mRNA delivered by lipid nanomaterials.
[0040] The lipid nanomaterial is a phospholipid nanoparticle. Preferably, the phospholipid nanoparticle is 1AP27, and the STING mRNA delivered by the lipid nanomaterial is 1AP27@STING mRNA.
[0041] See also Figure 2 The preparation method of the phospholipid nanocompound 1AP27 is as follows:
[0042] S1. 2-Chloro-1,3,2-dioxaphospholane-2-oxide (1.1 eq.) was slowly added to a beaker containing alcohol (1.0 eq.) and Et3N (1.1 eq.) and stirred at -15°C for 5 minutes. Afterwards, the reaction was continued at 25°C for 12 hours. The precipitate was filtered and rinsed with ether three times. The mixture was filtered to remove triethylamine hydrochloride and the filtrate was concentrated by rotary evaporation to obtain the alkylated dioxaphospholane molecule. The yields of all syntheses were over 90%.
[0043] S2. In a round-bottom flask, the alkylated dioxaphospholane molecule (1.0 equivalent) and the amine (1.0 equivalent) were dissolved into CH3CN. The resulting mixture was heated to 70°C and stirred for 3d-5d. The reaction was monitored by TLC analysis and fractionated on a silica gel column with a solvent gradient of 4% methanol in dichloromethane to 20% methanol in dichloromethane. The final ionizable phospholipids were concentrated by rotary evaporation and dried under vacuum for 24h.
[0044] The preparation method of the 1AP27@STING mRNA is as follows:
[0045] Lipid nanomaterials were synthesized for the delivery of STING mRNA according to the molar ratio of phospholipid nanocompound (1AP27): dioleoyl L-α-phosphatidylethanolamine (DOPE): cholesterol (Cholesterol): 1,2-dimyristoyl-RAC-glycero-3-methoxypolyethylene glycol 2000 (DMG-PEG2000) = 50:40:20:0.25.
[0046] Example 1: Preparation of MSA-2-Cu:
[0047] 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;
[0048] S2: Weigh 16.0 mg (0.1 mmol) of copper sulfate, dissolve it in purified water and add it to the solution in 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 a compound, which is recorded as MSA-2-Cu.
[0049] The characterization data of the nuclear magnetic resonance hydrogen spectrum of the MSA-2-Cu are as follows: 1 H NMR (400MHz, DMSO) δ8.32(s,2H),7.64(s,2H),7.60(s,2H),3.88(s,6H),3.83(s,6H),2.68-2.66(m,4H),2.33-2.32(m,4H).
[0050] 13 C NMR (100MHz, DMSO) δ183.16,151.62,151.23,149.09,148.97,144.51,142.09,141.92,139.41,136.91,136.07,13 3.09,132.98,131.83,130.84,130.80,107.11,107.02,104.79,104.73,56.38,56.31,56.30,56.06,33.58,33.15.
[0051] IR(KBr,ν,cm -1 ):2938,2836,1651,1510,1432,1360,1255,1059,987,645;
[0052] HRMS(ESI-TOF):m / z calcd for:C 28 H 27 CuO 10 S2,650.0341[M+H] + ; found:650.0380.
[0053] Example 2: Synthesis of ionizable phospholipid compounds
[0054] See also Figure 2 , synthesized 81 ionizable phospholipids with chemical formulas and synthesis pathways such as Figure 2 shown.
[0055] S1. 2-Chloro-1,3,2-dioxaphospholane-2-oxide (1.1 eq.) was slowly added to a beaker containing alcohol (1.0 eq.) and Et3N (1.1 eq.) and stirred at -15°C for 5 minutes. Afterwards, the reaction was continued at 25°C for 12 hours. The precipitate was filtered and rinsed with ether three times. The mixture was filtered to remove triethylamine hydrochloride and the filtrate was concentrated by rotary evaporation to obtain the alkylated dioxaphospholane molecule. The yields of all syntheses were over 90%.
[0056] S2. In a round-bottom flask, the alkylated dioxaphospholane molecule (1.0 equivalent) and the amine (1.0 equivalent) were dissolved into CH3CN. The resulting mixture was heated to 70°C and stirred for 3d-5d. The reaction was monitored by TLC analysis and fractionated on a silica gel column with a solvent gradient of 4% methanol in dichloromethane to 20% methanol in dichloromethane. The final ionizable phospholipids were concentrated by rotary evaporation and dried under vacuum for 24h.
[0057] Example 3: Optimization of phospholipid nanoparticles
[0058] MC38 cells were seeded at 5×10 4 24-well plates were filled with 10 cells / well. After 24 hours, MC38 cells were treated with Control (PBS) and 34 phospholipid nanoparticles (81 phospholipid nanoparticles in total) were used to deliver GFP plasmid. After 24 hours, cells were fixed with PFA, cell nuclei were detected with DAPI staining, and cells were captured using a fluorescence microscope. Figure 3 , 1AP27 phospholipid nanoparticles have the characteristics of low cytotoxicity and high transfection efficiency.
[0059] Example 4.1 Synthesis and preparation of AP27@STING mRNA
[0060] Before starting, ensure that all supplies, reagents, and working environment are RNase-free. STING mRNA (lipid / mRNA = 10 / 1, mass ratio), phospholipid nanocompound (1AP27): dioleoyl L-α-phosphatidylethanolamine (DOPE): cholesterol (Cholesterol): 1,2-dimyristoyl-RAC-glycero-3-methoxypolyethylene glycol 2000 (DMG-PEG2000) = 50:40:20:0.25 molar ratio preparation. Mix the ethanol lipid mixture with the acidic aqueous buffer containing the oligonucleotide, refer to Figure 1 A ratio of 1:3 of ethanol lipid mixture to water buffer was used, and the mixing process used an automated microfluidic mixing device, resulting in 1 ml of 1AP27@STING mRNA as the final product.
[0061] Example 5: Verification of the in vitro anti-tumor effect of MSA-2-Cu
[0062] Cell viability assay
[0063] MC38 and B16F10 cells were seeded into 96-well plates (10 cells per well). 4 cells), and 24 hours later, MC38 cells were treated with Control (DMSO), 25, 50, 75, 100, 200, 300 μM MSA-2-Cu. MC38 cells were treated with Control (DMSO), 37.5, 75, 150 μM MSA-2-Cu. After 24 hours, 10 μL CCK8 test solution was added to each well, incubated at 37°C for 1 hour, and the absorbance at a wavelength of 450 nm was measured using an ELISA reader. Please refer to Figure 4 B, 4C, MSA-2-Cu significantly induced cell death in MC38 cells and B16F10 cells at 50 μM in a dose-dependent manner. However, MSA-2 had no obvious toxicity to MC38 cells and B16F10 cells in vitro.
[0064] B16F10 cells were seeded into 24-well plates at 1 × 10 cells per well. 5 After 24 h, B16F10 cells were treated with DMSO, 75 μM MSA-2 and MSA-2-Cu for 12 and 24 h. Cell images were taken under an inverted microscope. Figure 4 D, Images show that MSA-2-Cu can induce cell death.
[0065] 5.2. Western Blot method to detect changes in protein content:
[0066] See also Figure 5, treated the mouse macrophage cell line RAW264.7 with different concentrations of MSA-2 and MSA-2-Cu. It was found that MSA-2-Cu could induce the phosphorylation of STING (P-STING) and increase the phosphorylation of TBK1 (P-TBK1). The results showed that MSA-Cu could activate the STING pathway similar to MSA-2.
[0067] Example 6: Verification of the in vitro anti-tumor effect of MSA-2-Cu combined with lipid nanomaterial STING mRNA
[0068] Female C57BL / 6 mice were subcutaneously inoculated with B16F10 cells (2×10 5 When the tumor size is about 100 mm 3 At the same time, the mice were divided into 4 groups, control group, MSA-2 group, MSA-2-Cu group and 1AP27@STING mRNA+MSA-2-Cu group. The mice were injected intratumorally with drugs three times at intervals of 2 days and 4 days. The MSA-2 and MSA-2-Cu groups were treated with 150μg MSA-2 or MSA-2-Cu, and the 1AP27@STING mRNA+MSA-2-Cu group was injected with 3μg 1AP27@STING mRNA, and 150μg MS A-2-Cu was injected 6 hours later. The control group was treated with the same volume of drug solvent, and the tumor size was measured with a digital caliper every 3 days and its area was calculated (length x width x width / 2). The mice were killed when the tumor diameter reached 1.6 cm. After the injection, the mice were cultured in the same environment and the survival rate and tumor volume of the mice were observed.
[0069] See also Figure 6 BC, tumor growth was significantly reduced after mice were treated with MSA-2, MSA-2-Cu, and 1AP27@STING mRNA+MSA-2-Cu ( Figure 6 B, C), especially the 1AP27@STING mRNA+MSA-2-Cu group.
[0070] See also Figure 6 D, The survival time of mice in the 1AP27@STING mRNA+MSA-2-Cu group was significantly prolonged compared with that in other groups.
[0071] Example 7: Study on the anti-tumor mechanism of 1AP27@STING mRNA+MSA-2-Cu:
[0072] Female C57BL / 6 mice were subcutaneously inoculated with B16F10 cells (2×10 5 / mouse). The tumor size was approximately 100 mm 3,The mice were divided into 4 groups: control group, MSA-2 group, MSA-2-Cu group and 1AP27@STING mRNA+MSA-2-Cu group. One day after the last administration, the mice were anesthetized and orbital blood was taken, and the mice were killed to extract tumor tissues.
[0073] 7.1. Detection of immune factors IFN-β and IL-6:
[0074] After anesthesia, blood was collected from the mouse's orbits, and the blood was centrifuged after standing at room temperature for 4 hours. The supernatant was used to detect the level of INF-β in serum using a mouse IFN-β ELISA Kit. The level of IL-6 in serum was detected using a mouse IFN-β ELISA Kit.
[0075] See also Figure 7 B, After treatment with MSA-2, MSA-2-Cu, and 1AP27@STIN mRNA + MSA-2-Cu, the proinflammatory cytokines IFN-β and IL-6 in serum increased, especially 1AP27@STING mRNA + MSA-2-Cu treatment induced increased levels of IFN-β and IL-6.
[0076] 7.2. Histological Analysis:
[0077] Mouse tumors were fixed with MDF fixative, then embedded in paraffin, sectioned, and stained with a hematoxylin-eosin (H&E) staining kit. Images were obtained using an optical microscope. Figure 7 C, Histomorphology of tumors showed that 1AP27@STING mRNA+MSA-2-Cu treatment induced more cell death than other groups.
[0078] 7.3. Immunofluorescence staining:
[0079] (1) Mouse tumor tissue was fixed with MDF fixative, paraffin embedded, and sectioned. (2) Paraffin was removed and tissue was rehydrated. (3) After antigen retrieval, the tissue was permeabilized with 0.1% TritonX-100. (4) The blocking reagent was blocked with 5% BSA at room temperature for 1 hour. Primary antibody staining: CD4 antibody, CD8 antibody. Secondary antibody: Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody was placed at room temperature for 2 hours. Cell nuclei were stained with DAPI. Images were obtained using a fluorescence microscope.
[0080] See also Figure 7D, CD8+ T cells in MSA-2-Cu and 1AP27@STING mRNA+MSA-2-Cu treatments were effectively activated. CD4+ T cells treated with 1AP27@STING mRNA+MSA-2-Cu were slightly activated compared with the control group, MSA-2, and MSA2-Cu.
[0081] In summary, two strategies for improving the anti-tumor therapy of MSA-2 are adopted in this application. First, copper-modified MSA-2 is synthesized in this application, and the method shows that the cytotoxicity of Cu enhances the STING pathway. 81 ionizable phospholipid nanoparticles are also synthesized in this application, among which the phospholipid nanoparticles 1AP27@STING mRNA can effectively deliver STIING-mRNA and increase the expression of STING protein. Combining 1AP27@STING mRNA with MSA-2-Cu can significantly reduce the growth of B16F10 tumors and increase the survival rate of mice. The 1AP27@STING mRNA+MSA-2-Cu treatment in this application stimulates a strong immune response by activating the cGAS-STING pathway, improves the anti-tumor effect of MSA-2, and provides a promising strategy for enhancing tumor immunotherapy.
Claims
1. Application of MSA-2-Cu in the preparation of tumor treatment products, Features: The MSA-2-Cu is the copper salt of MSA-2.
2. Use of the MSA-2-Cu according to claim 1 in preparing a product for treating tumors, Features: The MSA-2-Cu releases Cu and MSA-2 at the tumor site, and efficiently activates the cGAS-STING signaling pathway through Cu, while MSA-2 can selectively target tumor tissues and show a higher ability to activate the STING pathway.
3. Use of the MSA-2-Cu according to claim 1 in preparing a product for treating tumors, Features The chemical formula of MSA-2-Cu is as follows:
4. Use of the MSA-2-Cu according to claim 1 in preparing a product for treating tumors, Features: The synthesis path of the MSA-2-Cu is as follows:
5. Use of the MSA-2-Cu according to claim 1 in preparing a product for treating tumors, Features: The specific preparation method of the MSA-2-Cu is as follows: S1: 4-(5,6-dimethoxybenzo[B]thiophen-2-yl)-4-oxobutanoic acid and sodium hydroxide were placed in a reactor, purified water was added, and the reaction was carried out at room temperature for 12 hours. The reaction was stopped and the reaction was not further processed and was reserved for the next step; S2: Take copper sulfate, dissolve it in purified water and add it to the solution in the first step. Let it 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 a compound, recorded as MSA-2-Cu.
6. Use of the MSA-2-Cu according to claim 1 in preparing a product for treating tumors, Features: The product also includes STING mRNA delivered by lipid nanomaterials, wherein the lipid nanomaterials are phospholipid nanoparticles.
7. Use of the MSA-2-Cu according to claim 6 in preparing a product for treating tumors, Features: The phospholipid nanocompound is 1AP27, and the STING mRNA delivered by the lipid nanoparticles is 1AP27@STING mRNA.
8. Use of the MSA-2-Cu according to claim 7 in preparing a product for treating tumors, Features: The preparation method of 1AP27 is as follows: S1. 2-chloro-1,3,2-dioxaphospholane-2-oxide was slowly added to a beaker containing alcohol and Et3N and stirred at -15°C for 5 minutes; Afterwards, the reaction was continued at 25°C for 12 hours; The precipitate was filtered and rinsed three times with ether; the mixture was filtered to remove triethylamine hydrochloride, and the filtrate was concentrated by rotary evaporation to obtain the alkylated dioxaphospholane molecule; the yield of all syntheses exceeded 90%; S2. In a round-bottom flask, dissolve the alkylated dioxaphospholane molecules and amine into CH3CN; The resulting mixture was heated to 70°C and stirred for 3d-5d; the reaction was monitored by TLC analysis and fractionated on a silica gel column with a solvent gradient of 4% methanol in dichloromethane to 20% methanol in dichloromethane; the final ionizable phospholipids were concentrated by rotary evaporation and dried under vacuum for 24h.
9. Use of the MSA-2-Cu according to claim 8 in preparing a product for treating tumors, Features: The 1AP27@STING mRNA lipid nanoparticles are synthesized according to the molar ratio of phospholipid nanocompound (1AP27): dioleoyl L-α-phosphatidylethanolamine (DOPE): cholesterol (Cholesterol): 1,2-dimyristoyl-RAC-glycero-3-methoxypolyethylene glycol 2000 (DMG-PEG2000) = 50:40:20:0.25 for the delivery of STING mRNA.