Application of resibufogenin in preparation of STING inhibitor

Abstract

The invention relates to an application of resibufogenin in preparation of an STING inhibitor. The invention creatively discovers that resibufogenin can be combined with STING in a targeted manner to inhibit inflammatory response. According to the application, it is found through animals that resibufogenin can quickly act on the heart after being orally taken, immune response is relieved through targeting STING, the protection effect on the non-infarction area of the heart is achieved, myocardial cell damage of a heart failure model after myocardial infarction can be relieved, inflammatory damage and tissue fibrosis can be relieved, and then the heart function and heart failure are improved. The invention provides a theoretical basis for researching the treatment strategy of the heart failure, and provides an embedding point for preparing a novel medicine for preventing or treating the heart failure. Meanwhile, the invention develops a new pharmaceutical application of resibufogenin, and provides a new strategy for development and utilization of resibufogenin.

Description

[0001] This application claims priority to a Chinese patent application filed on February 14, 2025, with application number 2025101667044, and invention name “Application of ester toad venom in the preparation of STING inhibitors”, the entire contents of which are incorporated by reference into this application. Technical Field

[0002] The present invention belongs to the technical field of biomedicine, and particularly relates to the application of bufotoxin in the preparation of a STING inhibitor. Background Art

[0003] Heart failure is the terminal stage of various heart diseases. Inflammation plays an important role in the pathogenesis of heart failure, but the results of targeted inflammatory treatment in clinical trials of heart failure are not optimistic. For example, a multicenter controlled trial of rosuvastatin for the treatment of heart failure showed that the high-sensitivity CRP of patients decreased after drug treatment, but the mortality rate of elderly patients with heart failure did not decrease. At the same time, some trials using anti-TNF drugs were abandoned due to ineffectiveness and harm to the intervention group. Drugs targeting inflammation in heart failure are urgently needed to be developed.

[0004] The stimulator of interferon genes (STING) is a key innate immune signal transduction protein that is mainly present in the endoplasmic reticulum. It is activated when it recognizes pathogen DNA (such as DNA of viruses, bacteria, etc.) in the cytoplasm or abnormal release of its own DNA (such as after cell damage). The activated STING will initiate a series of signal transductions, ultimately inducing the production of type I interferon and other proinflammatory cytokines, playing an important role in the body's antiviral and antibacterial immune responses and the occurrence and development of autoimmune diseases.

[0005] Therefore, STING inhibitors play an important role in the treatment of inflammatory and autoimmune diseases. Currently, nitrofuran derivatives such as H-151, C-178 and C-176 are highly efficient and selective small molecule inhibitors targeting STING. Studies have shown that C-178 specifically binds to mouse STING (mmSTING) and has a good inhibitory effect on this pathway. The recently developed inhibitor H-151 (indole urea) antagonizes STING palmitoylation and reaches therapeutic drug concentrations in mice. However, its toxicity is high, and its effect on the immune inflammatory response in the onset of heart failure is unclear, so it cannot be used as a potential drug for the treatment of heart failure.

[0006] Esterbufagin is a component of bufadienolide, a cardiac glycoside compound that can exert cardiotonic and anti-inflammatory effects. It is unclear whether esterbufagin can bind to STING and act as a targeted inhibitor of STING. At the same time, its therapeutic effect in heart failure is also unclear. Summary of the invention

[0007] In view of the deficiencies in the prior art, the object of the present invention is to provide a use of bufotoxin in the preparation of a STING inhibitor.

[0008] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0009] In a first aspect, the present invention provides the use of bufotoxin in the preparation of a STING inhibitor.

[0010] Esterbufagin is a component of bufadienolide, a cardiac glycoside compound (a class of steroids and their sugar ligands that can enhance myocardial contraction). The structure is as follows:

[0011]

[0012] The unsaturated lactone ring at the β position of C-17 is an important pharmacophore of cardiac glycosides. The unsaturated lactone ring at the C-17 position of cardiac glycosides can act as a hydrogen bond receptor to form hydrogen bonds with glutamine (Gln111) and asparagine (Asn122) on the transmembrane domain TM1-TM2 of the α subunit of sodium-potassium ATPase. The unsaturated lactone ring at the C-17 position may be the key pharmacophore for cardiac glycosides to exert cardiotonic and anti-inflammatory effects.

[0013] The present invention creatively discovered that ester toad venom can target and bind to STING to inhibit inflammatory response. The present invention discovered through animals that ester toad venom can act quickly on the heart by oral administration, and by targeting STING, it reduces the immune response, has a protective effect on the non-infarcted area of ​​the heart, can reduce myocardial cell damage in the heart failure model after myocardial infarction, reduce inflammatory damage and tissue fibrosis, and then improve cardiac function and heart failure. The present invention provides a theoretical basis for studying the treatment strategy of heart failure and provides an embedding point for the preparation of new drugs for preventing or treating heart failure. At the same time, the present invention develops a new drug use for ester toad venom, providing a new strategy for the development and utilization of ester toad venom.

[0014] According to the research results of the present invention, the ester toad venom can target and bind to STING and inhibit the related functions of the STING pathway. Therefore, the results show that ester toad venom can be used as an in vitro experimental reagent in the field of scientific research, such as exploring the STING activation mechanism, the STING signal transduction pathway, and diseases related to the activation of the STING signaling pathway. The STING inhibitor claimed in the present invention is not used to eliminate the cause or lesion, that is, it is an application in the preparation of STING inhibitors for non-therapeutic purposes.

[0015] In a second aspect, the present invention provides the use of bispyribac in the preparation of a drug for inhibiting the activation of the STING signaling pathway.

[0016] In a third aspect, the present invention provides the use of siritinib in the preparation of a medicament for preventing or treating a disease associated with activation of the STING signaling pathway.

[0017] In a fourth aspect, the present invention provides the use of bispyribac in the preparation of a medicament for preventing or treating a STING-mediated disease.

[0018] Preferably, the STING-mediated disease comprises an inflammatory disease or an autoimmune disease.

[0019] In a fifth aspect, the present invention provides the use of bispyribac in the preparation of a medicament for preventing or treating heart failure.

[0020] Preferably, the drug further comprises a pharmaceutically acceptable excipient.

[0021] Preferably, the pharmaceutically acceptable excipients include any one or a combination of at least two of a carrier, a diluent, a filler, a binder, a wetting agent, a disintegrant, an emulsifier, a solubilizer, a solubilizing agent, an osmotic pressure regulator, a surfactant, a coating material, a colorant, a pH regulator, an antioxidant, an antibacterial agent or a buffer.

[0022] Preferably, the dosage form of the drug is any pharmaceutically acceptable dosage form.

[0023] Preferably, the dosage form of the drug includes inhalation, injection, aerosol, nebulizer, granule, tablet, capsule, solution or granule.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] The present invention creatively discovered that ester toad venom can target and bind to STING to inhibit inflammatory response. The present invention discovered through animals that ester toad venom can act quickly on the heart by oral administration, and by targeting STING, it reduces the immune response, has a protective effect on the non-infarcted area of ​​the heart, can reduce myocardial cell damage in the heart failure model after myocardial infarction, reduce inflammatory damage and tissue fibrosis, and then improve cardiac function and heart failure. The present invention provides a theoretical basis for studying the treatment strategy of heart failure and provides an embedding point for the preparation of new drugs for preventing or treating heart failure. At the same time, the present invention develops a new drug use for ester toad venom, providing a new strategy for the development and utilization of ester toad venom. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 is the immunoblot image of STING protein and β-actin protein;

[0027] Figure 2 is the relative band intensity diagram of STING protein after treatment with bufotoxin and DMSO;

[0028] Figure 3 is the relative band intensity of β-actin protein after treatment with bufotoxin and DMSO;

[0029] Figure 4 This is the distribution map of bufotoxin in mouse heart tissue;

[0030] Figure 5 are echocardiograms of mice in different treatment groups;

[0031] Figure 6 It is the test result diagram of left ventricular ejection fraction (EF) of mice in different treatment groups;

[0032] Figure 7 It is the test result of the short axis shortening rate (FS) of mice in different treatment groups;

[0033] Figure 8 It is the test result diagram of the thickness of the interventricular septum (IVSs) of mice in different treatment groups;

[0034] Fig. 9 It is the test result of left ventricular end-systolic posterior wall thickness (LVPWs) in mice of different treatment groups;

[0035] Fig.10 It is the test result of left ventricular end-diastolic posterior wall thickness (LVPWd) in mice of different treatment groups;

[0036] Fig.11 It is the test result diagram of left ventricular end-systolic volume (LVESV) of mice in different treatment groups;

[0037] Fig.12 It is the test result of the end-diastolic thickness of the ventricular septum (IVSd) of mice in different treatment groups;

[0038] Fig.13 is the pathological staining picture of heart tissues of mice in different treatment groups;

[0039] Fig.14 is a graph of LDH expression levels in mice of different treatment groups;

[0040] Fig.15 It is a graph of the mRNA expression level of STING in mice in different treatment groups;

[0041] Fig.16 is a graph of the mRNA expression levels of TNF-α in mice in different treatment groups;

[0042] Fig.17 is the graph of NF-κB mRNA expression levels in mice of different treatment groups;

[0043] Fig.18is the graph of mRNA expression levels of TBK in mice in different treatment groups;

[0044] Fig.19 is the graph of mRNA expression levels of Tgf-β in mice in different treatment groups;

[0045] Fig. 20 is the graph of mRNA expression levels of α-SMA in mice in different treatment groups;

[0046] Fig.21 is the graph of colla1 mRNA expression levels in mice of different treatment groups;

[0047] Fig. 22 It is a graph of the mRNA expression level of IL-6 in mice under different treatment groups. DETAILED DESCRIPTION

[0048] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0049] In the following examples, unless otherwise specified, the reagents and consumables used were purchased from conventional reagent manufacturers in the field; unless otherwise specified, the experimental methods and technical means used were conventional methods and means in the field.

[0050] Example 1

[0051] In this example, the targeting of bufotoxin to STING was studied.

[0052] The binding of sediptoxin to STING protein was analyzed by using the Cell Thermal Shift Assay (CETSA) technique. When STING binds to sediptoxin, its thermal stability changes. By measuring this change, the interaction between the drug and the protein is identified, and the binding of the drug and the protein is determined.

[0053] Experimental methods:

[0054] 1) RAW cells were cultured first using 1640 medium containing 10% FBS and 1% penicillin-streptomycin (6 mL medium per culture dish). When the cell density reached 80%-90%, the cells were divided into DMSO group and 240 (40 um) group, and 6 μL DMSO and 6 μL bufotoxin stock solution (40 μM) were given for intervention for 24 h, respectively.

[0055] 2) After the intervention, transfer the cell reaction solution to two 50 mL centrifuge tubes, centrifuge at 4°C, 300 × g for 5 min to obtain cell pellets, wash the pellets twice with 20 mL PBS, and centrifuge at 300 × g for 5 min. Resuspend the cell pellets with 0.8 mL ice-cold PBS containing protease inhibitors. Take 80 μL of each and dispense into 10 0.2 mL PCR tubes (approximately 3.4 × 10 6 / tube), 10 temperatures each for the 240 group and the DMSO group (37℃, 41℃, 44℃, 47℃, 50℃, 53℃, 56℃, 59℃, 63℃, 67℃), a total of 20 tubes.

[0056] 3) Heat treatment of cell suspension, set 10 temperature points (37℃, 41℃, 44℃, 47℃, 50℃, 53℃, 56℃, 59℃, 63℃, 67℃) in the PCR instrument, heat the PCR tube in the PCR instrument for 3 minutes, and then put it on ice. Add 20μL of lysis buffer containing protease inhibitors to each sample for 20 minutes. Oscillate and mix, transfer the cell lysate to a 1.5mL centrifuge tube, centrifuge at 4℃, 20000×g for 20 minutes, place the sample on ice after centrifugation, and take the supernatant. Take 40μL of sample and add 10μL 5×SDS loading buffer, heat at 99℃ for 10 minutes. Centrifuge at 4℃, 20000×g for 20 minutes, and take the supernatant. Perform western blot.

[0057] Experimental results:

[0058] like Figure 1-3 As shown in the figure, after 40μM sedifugine intervened in RAW cells for 24h, the STING protein had a corresponding binding with sedifugine, and compared with the DMSO group, the stability at different temperatures was increased, especially at 47℃, 50℃, and 53℃. There was no statistical difference in the changes of β-actin in the control group, proving that sedifugine can bind to STING.

[0059] Example 2

[0060] In this example, the pharmacokinetics of bufotoxin were studied.

[0061] Experimental methods:

[0062] Grouping: Twelve C57 mice were divided into four groups, corresponding to four time points: 5min, 15min, 30min, and 45min, with 3 mice in each group.

[0063] Collection of heart samples: 5min, 15min, 30min and 45min after mice were gavaged with 30mg / kg toad venom solution, the mice were killed at different time points, the hearts were taken out, excess tissues were cut off, residual blood was drained, physiological saline 3 times the volume of the heart weight was added, and the heart was placed in a homogenizer for homogenization. After the homogenization was completed, the heart was centrifuged at 3000rpm for 15min, and the supernatant was transferred to another centrifuge tube.

[0064] After adding 3 times the volume of methanol solution to the homogenate supernatant, centrifuge at 12000rpm for 10min, aspirate the supernatant, concentrate in a centrifugal concentrator, re-dissolve the residue with 100μL of 80% methanol solution, centrifuge at 12000rpm for 15min at 4°C, take 50L of supernatant for LC-MS / MS analysis, record the chromatogram, and analyze the pharmacokinetics of cardiac tissue after administration.

[0065] Experimental results:

[0066] The results are as follows Figure 4 As shown, after oral administration, bufotoxin can be rapidly distributed to the heart tissue within 5 minutes to exert its effect.

[0067] Example 3

[0068] In this example, the efficacy of thiabendazole in improving heart failure was studied. A mouse model of heart failure after myocardial infarction was established, and thiabendazole and fosinopril were administered for intervention. The efficacy of thiabendazole in improving heart failure was observed by echocardiography, serum myocardial injury marker detection, and QPCR experiments.

[0069] Construction of animal model:

[0070] Animal strain: C57 mice (8 weeks old, 25-30 g)

[0071] Grouping: Sham (sham operation group), Model (model group), 240 group, Fosinprill (fosinpril group), 7 rats in each group. Among them, the model group, 240 group, and fosinpril group were divided into groups according to EF% (ejection fraction) after echocardiographic evaluation.

[0072] Administration: 240 and fosinopril were prepared with 5% sodium carboxymethylcellulose solution (prepared with pure water). The sham operation group and the model group were gavaged with 0.1 mL / 10 g of 5% sodium carboxymethylcellulose solution, the 240 group was gavaged with 0.1 mL / 10 g of bufotoxin suspension, and the fosinopril group was gavaged with 0.1 mL / 10 g of fosinopril suspension. The administration was continued for 7 days, once a day.

[0073] Modeling: After 1 week of adaptive feeding of mice, 0.15 mL / 10 g of Avertin was injected into the experimental mice intraperitoneally to maintain anesthesia. The mice were fixed on the operating table and the hair on the left chest of the mice was removed. The endotracheal tube was intubated and connected to the ventilator (the respiratory rate was set to 100 times / min and the tidal volume was 2 mL). Using the surface landmarks, the skin and muscle were cut open between the 4th and 5th ribs and the incision was about 5-6 mm long. The intercostal muscles were bluntly separated to expose the heart. The left anterior descending branch of the coronary artery was found and the anterior descending branch was ligated with a surgical needle and a 7-0 thread (the sham operation group did not perform ligation, and only the needle suture was passed through the heart). The muscles and ribs were sutured with a 5-0 thread first, and then the skin was sutured. An electrocardiogram was performed immediately after surgery to determine whether the model was successful. After 24 hours, an echocardiogram was performed to determine the cardiac function.

[0074] 3.1 Echocardiography

[0075] Test method:

[0076] Transthoracic color Doppler ultrasound examination was performed on mice in each group. Ultrasound transsternal left heart long axis section was used to measure the thickness of the interventricular septum (IVSs), left ventricular end-diastolic volume (LVESv), left ventricular end-diastolic posterior wall thickness (LVPWd), left ventricular end-systolic posterior wall thickness (LVPWs), left ventricular end-diastolic diameter (LVIDd), left ventricular end-systolic diameter interventricular septum thickness (IVSd), left ventricular ejection fraction (EF) and fractional shortening (FS).

[0077] Experimental results:

[0078] like Figure 5-12 As shown, compared with the model group, the contractile function and wall thickness of the left ventricle were improved, indicating that bispyribac can effectively improve cardiac function.

[0079] 3.2 Effects of bufotoxin on pathological parameters of post-MI heart failure mice

[0080] Test method:

[0081] The cardiac tissue in the infarct edge area was subjected to pathological staining, including HE staining, Masson staining, and WGA staining.

[0082] 1) HE staining:

[0083] Dewaxing to water: Dewaxing tissue sections with xylene for 5 minutes, twice. Then, gradient hydration was performed with anhydrous ethanol, 95% ethanol, 90% ethanol, 80% ethanol, and 70% ethanol for 2-3 minutes each, and finally, rinsed with distilled water for 3 minutes.

[0084] Hematoxylin staining: Hematoxylin staining for 15 minutes.

[0085] Color separation and blueing: Use 1.0% hydrochloric acid alcohol for color separation for 10 seconds, and quickly wash with distilled water; use light ammonia water for blueing, and wash with distilled water. Check under a microscope while differentiating until the cell nucleus is stained appropriately.

[0086] Rinse with running water: Rinse with running water for 3 minutes to wash away the alkaline water.

[0087] Eosin staining: 1% eosin staining for 8 min.

[0088] Dehydration: quick wash with distilled water, dehydration with alcohol in ascending steps, quick wash with 70% ethanol, 80% ethanol and 90% ethanol, 45s with 95% ethanol, microscopic examination; 3min with 100% alcohol, 2 times; 5min with xylene, 2 times.

[0089] Sealing: Neutral gum sealing.

[0090] Observe under the microscope.

[0091] 2) Masson staining:

[0092] Dewaxing to water: Place the paraffin sections in xylene, anhydrous ethanol, 90% ethanol, 80% ethanol, and 70% ethanol in sequence, and finally rinse with distilled water.

[0093] Mordant treatment: Immerse the sections in Bouin solution, place in a 37°C incubator for accelerated treatment for 2 hours, and then rinse with running water until the yellow color completely fades.

[0094] Azure blue staining: Use Azure blue staining solution to drip stain on the sections for 3 minutes to ensure uniform coloring, then gently rinse with distilled water to remove excess stain.

[0095] Mayer's hematoxylin counterstaining: Add Mayer's hematoxylin staining solution for 3 minutes to enhance the contrast of tissue structure. Wash lightly with distilled water to remove floating color.

[0096] Acid ethanol differentiation: Place the sections in acid ethanol differentiation solution for a few seconds to fine-tune the depth of hematoxylin staining. Rinse the sections with distilled water for about 10 minutes to ensure that the differentiation solution is completely removed.

[0097] Ponceau fuchsin staining: Add ponceau fuchsin staining solution for 10 minutes to highlight specific tissue structures. Gently rinse the sections with distilled water to remove excess staining solution.

[0098] Phosphomolybdic acid treatment: Place the sections in phosphomolybdic acid solution for about 10 minutes to enhance the contrast of subsequent staining.

[0099] Aniline blue staining: After pouring off the phosphomolybdic acid solution, add aniline blue staining solution directly and stain for 5 minutes to further enrich the color of the tissue. Treat the sections with a weak acid solution for 2 minutes to optimize the staining effect.

[0100] Dehydration and transparency: First, the sections were rapidly dehydrated with 95% ethanol, followed by three dehydration treatments with anhydrous ethanol, each lasting 10 seconds. Finally, the sections were transparentized in xylene three times, each for 2 minutes.

[0101] Sealing: Finally, seal the sections with neutral gum to complete the Masson staining process.

[0102] Observe under the microscope.

[0103] WGA staining:

[0104] Fixation: Use 40% paraformaldehyde for fixation.

[0105] Permeabilization: The samples were treated with the permeabilizing agent Triton X-100 to increase the permeability of WGA.

[0106] Staining: WGA was labeled with Alexa Fluor 488 at a concentration of 10 μg / mL and incubated for 20 min.

[0107] Washing: Gently wash the samples with PBS buffer to remove unbound WGA.

[0108] Nuclear staining: DAPI was used to stain the cell nucleus so that the cell membrane and the cell nucleus could be observed simultaneously under a fluorescence microscope.

[0109] Sealing: Sealing the stained samples and preparing for microscopic observation.

[0110] Microscope observation: Observe using a fluorescence microscope.

[0111] Experimental results:

[0112] like Fig.13 As shown in the figure, it can be seen that bispyribac venom effectively reduces the infiltration of inflammatory cells in the infarct edge area and reduces myocardial fibrosis, indicating that bispyribac venom has a protective effect on the non-infarct area of ​​the heart.

[0113] 3.3 Effects of bufotoxin on myocardial damage in mice with heart failure

[0114] Test method:

[0115] When sampling, the eyeballs were removed and blood was collected. After standing at room temperature for two hours, the blood was centrifuged at 12000rpm for 15 minutes, and the serum after centrifugation was taken for subsequent testing.

[0116] The prepared serum samples were used to detect the serum myocardial injury marker LDH using a BS-460 fully automatic biochemical analyzer.

[0117] Experimental results: The results are as follows Fig.14 As shown, compared with the model group, the levels of serum myocardial injury markers decreased, indicating that bispyribac venom can alleviate myocardial cell damage in the heart failure model after myocardial infarction and improve cardiac function.

[0118] 3.4 Effects of bufotoxin on immune inflammation and myocardial fibrosis in mice with heart failure

[0119] Test method:

[0120] Sample preparation and RNA extraction: First, RNA needs to be extracted from the sample, and processes such as heart tissue lysis, RNA isolation and purification are performed to ensure the purity and integrity of the extracted RNA.

[0121] Take out the heart tissue from the -80℃ refrigerator, thaw on ice, weigh it, add 1mL of Trizol solution to every 25mg of tissue, put it into the homogenizer for homogenization, centrifuge at 3000rpm for 10min to get the homogenate supernatant. Add chloroform solution at a volume ratio of 5:1 of supernatant: chloroform solution. Invert the centrifuge tube repeatedly to mix, let it stand for 10min, and the solution will be layered. Then centrifuge at 12000rpm. After 15min, the layering is more obvious. Take the upper clear solution, add the same volume of isopropanol solution, invert repeatedly to mix, let it stand for 10min, and then centrifuge at 12000rpm. After 10min, white precipitate can be seen on the wall of the centrifuge tube. Discard the solution, add 200μL of anhydrous ethanol, mix well, centrifuge at 12000rpm for 5min, discard the anhydrous ethanol, and let it stand in the fume hood to dry.

[0122] Add 50 μL of enzyme-free water to each centrifuge tube and oscillate to mix. Use a UV spectrophotometer to measure the mRNA concentration. Use the EZB Color Reverse Transcription Kit (A0010CGQ) to perform reverse transcription reaction according to the instructions to convert the extracted RNA into cDNA.

[0123] PCR reaction system configuration: According to the experimental requirements, prepare a reaction system containing template, primers, fluorescent dye or probe, etc. Use 2× concentrated real-time qPCR MasterMix and make 3 parallel wells for each sample.

[0124] PCR amplification: Place the prepared reaction solution into the real-time qPCR instrument, set the corresponding temperature cycle and acquisition mode, perform amplification detection, and monitor the amount of PCR products in real time through changes in fluorescence signals.

[0125] Experimental results:

[0126] like Figure 15-22 As shown in the results, esterbufant can reduce inflammatory indicators such as TNFɑ, NF-κB, TGFβ, IL6, and reduce myocardial fibrosis indicators such as ɑ-SMA and Colla1, proving that esterbufant can reduce inflammatory damage, tissue fibrosis and improve cardiac function

[0127] The applicant declares that the present invention illustrates the technical solution of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0128] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0129] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. Application of ester toad venom in the preparation of STING inhibitors.

2. Use of sirenic venom in the preparation of drugs for inhibiting the activation of STING signaling pathway.

3. Use of sirenic venom in the preparation of drugs for preventing or treating diseases associated with activation of the STING signaling pathway.

4. Use of sirenotoxin in the preparation of drugs for preventing or treating STING-mediated diseases.

5. The use according to claim 4, wherein the STING-mediated disease comprises an inflammatory disease or an autoimmune disease.

6. Use of bufotoxin in the preparation of medicines for preventing or treating heart failure.

7. The use according to claims 2-6, characterized in that: The drug also includes pharmaceutically acceptable excipients.

8. The use according to claim 7, characterized in that: The pharmaceutically acceptable excipients include any one or a combination of at least two of carriers, diluents, fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, surfactants, coating materials, colorants, pH regulators, antioxidants, antibacterial agents or buffers.

9. The use according to claims 2-6, characterized in that: The dosage form of the drug is any pharmaceutically acceptable dosage form.

10. The use according to claims 2-6, characterized in that: The dosage form of the drug includes inhalation, injection, aerosol, nebulizer, granule, tablet, capsule, solution or granule.

Images