Application of Endosidin-2 in the preparation of antiviral drugs

Abstract

This invention belongs to the field of pharmaceutical technology, specifically relating to the application of Endosidin-2 in the preparation of antiviral drugs. This invention is the first to discover that the compound Endosidin-2 can effectively inhibit the replication of various viruses, including vesicular stomatitis virus, herpes simplex virus type I, bovine herpesvirus type I, and bovine parainfluenza virus type 3, blocking viral release, and exhibiting relatively low cytotoxicity. This indicates its potential for development into antiviral drugs against these viruses, thus opening up new pharmaceutical applications for Endosidin-2 and laying an experimental foundation and providing new perspectives for the development of highly effective and specific antiviral drugs. Therefore, it has significant practical application value.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to the application of Endosidin-2 in the preparation of antiviral drugs. Background Technology

[0002] The information disclosed in this background section is intended only to enhance understanding of the overall background of the invention and is not necessarily to be construed as an admission or in any way implying that such information constitutes prior art known to those skilled in the art.

[0003] Viruses are non-cellular organisms that parasitize host cells; their replication, transcription, and translation must all take place within the host cell. Infectious diseases caused by viruses are characterized by their wide variety, high infectivity, high morbidity, and high mortality, seriously endangering the lives and health of humans and animals. The development of drugs for viral diseases is both a challenge and a hot topic in global drug development. Developing safe and effective drugs and exploring new antiviral applications for existing drugs are crucial for advancing antiviral drug research and development, providing a reference for developing more effective antiviral drugs.

[0004] The small molecule drug Endosidin-2 (ES2, CAS: 1839524-44-5) has been found to improve the efficacy of cisplatin in clinical chemotherapy for epithelial ovarian cancer and can also be used to treat type 2 diabetes. Therefore, ES2 holds promise as a target drug for treating human diseases such as diabetes and cancer. However, the antiviral effects of ES2 remain unclear. Summary of the Invention

[0005] To address the shortcomings of the existing technologies, the inventors, through long-term technical and practical exploration, have developed an application of Endosidin-2 in the preparation of antiviral drugs. This invention, through research, has for the first time discovered and confirmed that Endosidin-2 has antiviral effects against vesicular stomatitis virus, herpes simplex virus type I, bovine herpesvirus type I, and bovine parainfluenza virus type 3, thus showing promise for development into antiviral drugs. Based on the above research results, this invention has been completed.

[0006] To achieve the above technical objectives, the present invention adopts the following technical solution:

[0007] A first aspect of the invention provides the use of Endosidin-2 in the preparation of antiviral products; wherein the virus includes at least one of vesicular stomatitis virus, herpes simplex virus type I, bovine herpesvirus type I, and bovine parainfluenza virus type 3. Therefore, Endosidin-2 is effective in preventing and / or treating diseases associated with the aforementioned viruses.

[0008] A second aspect of the present invention provides a pharmaceutical composition comprising Endosidin-2 with at least one other pharmaceutically active ingredient and / or at least one other non-pharmaceutical active ingredient.

[0009] A third aspect of the present invention provides a method for preventing and / or treating diseases related to the aforementioned viral infection, the method comprising administering the aforementioned Endosidin-2 or the aforementioned pharmaceutical composition to a subject.

[0010] Compared with existing technical solutions, one or more of the above technical solutions have the following beneficial effects:

[0011] The above-mentioned technical solution is the first to discover that the compound Endosidin-2 can effectively inhibit the replication of a variety of viruses, including vesicular stomatitis virus, herpes simplex virus type I, bovine herpesvirus type I, and bovine parainfluenza virus type 3, and block viral release. Moreover, it has relatively low cytotoxicity, indicating that it has the potential to be developed into an antiviral drug for the above-mentioned viruses. This opens up new drug applications for Endosidin-2 and lays an experimental foundation and provides a new perspective for the development of highly effective and specific antiviral drugs. Therefore, it has good practical application value. Attached Figure Description

[0012] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an improper limitation of the invention.

[0013] Figure 1 CC50 of Endosidin-2 in HeLa cells, Vero cells, and MDBK cells in this embodiment of the invention.

[0014] Figure 2 The IC50 values ​​of Endosidin-2 against VSV, HSV-1, BoHV-1 and BPIV3 viruses in this embodiment of the invention are shown.

[0015] Figure 3 The SI values ​​of Endosidin-2 applied to VSV, HSV-1, BoHV-1 and BPIV3 viruses in this embodiment of the invention are shown.

[0016] Figure 4 This invention demonstrates the effect of Endosidin-2 in inhibiting the replication of VSV, HSV-1, BoHV-1, and BPIV3 in this embodiment. Detailed Implementation

[0017] It should be noted that the following detailed descriptions are illustrative and intended to provide further explanation of this application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains.

[0018] It should be noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the exemplary embodiments according to this application. As used herein, the singular form is intended to include the plural form as well, unless the context clearly indicates otherwise. Furthermore, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, devices, components, and / or combinations thereof.

[0019] As mentioned earlier, there has been no discovery of the application of Endosidin-2 in antiviral applications.

[0020] In view of this, in a typical embodiment of the present invention, Endosidin-2 is used in the preparation of antiviral products. The virus includes at least one of vesicular stomatitis virus, herpes simplex virus type I, bovine herpesvirus type I, and bovine parainfluenza virus type 3. Therefore, Endosidin-2 is effective in preventing and / or treating diseases related to the aforementioned viruses.

[0021] It should be noted that this application is being made public for the first time and is not the same as its known medicinal uses.

[0022] The antiviral activity is at least characterized by the ability to directly inactivate the aforementioned viruses and / or inhibit their replication.

[0023] The product may be a drug or an experimental reagent, which is intended for use in basic research and can be used to construct relevant cell or animal models.

[0024] Therefore, in another specific embodiment of the present invention, the following is provided:

[0025] (a) Application of Endosidin-2 in the preparation of inactivated virus products;

[0026] (b) Application of Endosidin-2 in the preparation of products that inhibit viral replication.

[0027] The virus includes at least one of vesicular stomatitis virus, herpes simplex virus type I, bovine herpesvirus type I, and bovine parainfluenza virus type 3.

[0028] According to the present invention, not only is the use of Endosidin-2 in the preparation of antiviral drugs disclosed, but also that the effect can be enhanced when Endosidin-2 is administered in combination with at least one other pharmaceutically active ingredient. Endosidin-2 can also be used in combination with other non-pharmaceutical active ingredients as a substitute or supplement to other pharmaceutically active ingredients.

[0029] In another specific embodiment of the present invention, a pharmaceutical composition is provided, the pharmaceutical composition comprising Endosidin-2 and at least one other pharmaceutical active ingredient and / or at least one other non-pharmaceutical active ingredient.

[0030] The other active pharmaceutical ingredients include substances that inhibit and / or kill the aforementioned viruses.

[0031] The non-pharmaceutical active ingredients include pharmaceutically acceptable carriers, excipients, and / or diluents.

[0032] In another specific embodiment of the present invention, the non-pharmaceutical active ingredient includes:

[0033] Pharmaceutically compatible inorganic or organic acids or bases, polymers, block copolymers, monosaccharides, polysaccharides, ionic and nonionic surfactants or lipids;

[0034] Pharmacologically harmless salts, flavorings, vitamins, antioxidants, as well as stabilizers and / or preservatives.

[0035] The dosage forms of the pharmaceutical composition include: liquid dosage forms, solid dosage forms, topical preparations, and sprays;

[0036] In another specific embodiment of the present invention, the following dosage forms are included: true solutions, colloids, microparticles, emulsions, vortexes, tablets, capsules, pellets, aerosols, pills, powders, solutions, suspensions, emulsions, granules, suppositories, lyophilized powder injections, inclusion complexes, fillers, patches, and liniments.

[0037] In another specific embodiment of the present invention, a method for preventing and / or treating diseases related to the aforementioned virus is provided, the method comprising administering to a subject a therapeutically effective dose of Endosidin-2 or the aforementioned pharmaceutical composition.

[0038] The viruses include at least one of vesicular stomatitis virus, herpes simplex virus type I, bovine herpesvirus type I, and bovine parainfluenza virus type 3.

[0039] The subject refers to an animal that is already being treated, observed, or experimented on, preferably a mammal, and more preferably a ruminant, such as a cow. "Effective therapeutic dose" refers to the amount that leads to improvement in any parameter or clinical symptom. The actual dose may vary from subject to subject and does not necessarily refer to the total amount that eliminates all disease symptoms; it can be determined using methods known in the art, which will not be elaborated upon here.

[0040] The present invention will be further illustrated below with specific examples. These examples are for illustrative purposes only and do not limit the scope of the invention. Any simple modifications, equivalent variations, and alterations made to the embodiments based on the technical essence of the present invention shall fall within the scope of the present invention.

[0041] Example

[0042] 1. Materials

[0043] HeLa cells, Vero cells, MDBK cells, MDCK cells, and vesicular stomatitis virus (VSV), herpes simplex virus type I (HSV-1), bovine herpesvirus type I (BoHV-1), and bovine parainfluenza virus type 3 (BPIV3) were all preserved by the Ruminant Animal Disease Research Center of Shandong Normal University.

[0044] ES2 was purchased from SparkJade; DMSO was purchased from SolarBio; DMEM medium, PBS, fetal bovine serum and other cell culture reagents were all purchased from BI.

[0045] 2. Methods and Results

[0046] 2.1 Cytotoxicity assay of ES2

[0047] The CCK-8 assay kit was used to investigate the cytotoxicity of ES2 cells against HeLa, Vero, and MDBK cells. Cells that had grown into confluent monolayers were dispersed by trypsin digestion, counted, and 100 μL of the cell suspension was seeded into 96-well plates at 5 × 10⁻⁶ cells / well. 5 Cells per well. Cells were cultured in a 37℃, 5% CO2 incubator for 12-24 hours. After observing good cell adhesion, the culture medium was aspirated from each well. The experimental groups were added to DMEM medium containing different concentrations of ES2, while the control group was added to medium without ES2. The 96-well plates were then incubated in a 37℃, 5% CO2 incubator for 48 hours. 10 μL of CCK-8 solution was added directly to each well, and the plates were incubated in a 37℃, 5% CO2 incubator for 1 hour. The 96-well plates were then removed, and the OD value of each well at 450 nm was measured using a microplate reader. The data were analyzed, and cell viability was calculated: Cell viability % = (OD of drug-treated cells / OD of control cells) × 100%. The drug concentration (CC50) at T / C = 50% was determined, and a proliferation curve was plotted.

[0048] The results showed that the CC50 values ​​of ES2 in HeLa cells, Vero cells, and MDBK cells were 163.6 μM, 582.8 μM, and 314.6 μM, respectively. Figure 1 ).

[0049] 2.2 ES2 antiviral capability test

[0050] HeLa cells, Vero cells, and MDBK cells were passaged in 96-well culture plates, and VSV, HSV-1, BPIV3, and BoHV-1 (0.001 MOI / well) were seeded, respectively. The cells were incubated in a 37°C, 5% CO2 cell culture incubator for 1 h. Cell maintenance medium containing 2% serum with different concentrations of ES2 was added, and the cells were observed for 4-7 days. The IC50 and SI values ​​of ES2 on the virus were calculated.

[0051] The results showed that the IC50 values ​​of ES2 against VSV, HSV-1, BoHV-1, and BPIV3 were 73.7 μM, 142.3 μM, 66.82 μM, and 91.24 μM, respectively. Figure 2 The SI values ​​are all greater than 1, indicating that ES2 has the potential to resist the above four viruses. Figure 3 ).

[0052] 2.3ES2 inhibits viral replication

[0053] HeLa cells, Vero cells, and MDBK cells were passaged into 6-well culture plates. After reaching confluent monolayers, VSV, HSV-1, BoHV-1, and BPIV3 (0.1 MOI / well) were seeded, respectively. The cells were incubated at 37°C in a 5% CO2 incubator for 1 h. Cell maintenance media containing 2% serum (ES2 and DMSO, respectively) were added. After 24 h, the virus was harvested, and the TCID of the viral fluid was measured. 50 .

[0054] The results are as follows Figure 4 The results showed that the viral titers of VSV, HSV-1, BoHV-1 and BPIV3 were significantly lower after ES2 treatment than in the control DMSO treatment group, indicating that ES2 inhibited the replication of VSV, HSV-1, BoHV-1 and BPIV3.

[0055] The above description is merely a preferred embodiment of this application and is not intended to limit this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. Use of Endosidin-2 in the manufacture of an antiviral product; wherein, The virus is a bovine parainfluenza type 3 virus.

2. The use according to claim 1, characterized in that, The antiviral at least exhibits the ability to inactivate the virus and / or inhibit viral replication.

3. The use according to claim 1, characterized in that, The product is a pharmaceutical or a test reagent for use in basic research.

4. Use of Endosidin-2 for the preparation of a product for inactivating a virus, which is a bovine parainfluenza type 3 virus.

5. Use of Endosidin-2 for the preparation of a product for inhibiting viral replication, which is a bovine parainfluenza type 3 virus.

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