Application of interferon alpha combined with stiripentanol in virus infection resistance

Abstract

The invention relates to the technical field of medicines, and particularly discloses application of interferon alpha combined with stiripentanol in virus infection resistance. As an antiviral drug, interferon alpha has the problems of limited curative effect, remarkable side effect, narrow applicable crowd, inconvenience in medication and the like clinically. The invention finds that the accumulation of host lactic acid in the later stage of virus infection is a key negative factor, and lactic acid directly inhibits the antiviral activity of interferon alpha and induces inflammatory storm. Through combined use of a lactic dehydrogenase inhibitor stiripentol (Stiripentol, STP), lactic acid production is reduced, a lactic acid-mediated antiviral inhibition-inflammation promotion effect is reversed, and the antiviral activity of interferon alpha is enhanced and side effects are reduced. Experiments prove that the combined therapy can effectively inhibit virus replication and inflammatory factor expression, and a new strategy is provided for broad-spectrum antiviral therapy.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and more specifically, to the application of interferon-α combined with stethatisol in the treatment of viral infections. Background Technology

[0002] Interferon-alpha, an important antiviral drug, is mainly used clinically for the treatment of chronic hepatitis B and C. Its main molecular mechanism involves activating the JAK-STAT signaling pathway to regulate the body's immunity and enhance the immune cells' ability to clear virus-infected hepatocytes. However, its antiviral application has significant limitations: efficacy is affected by viral genotype and baseline viral load, with a response rate of only 30%-50% in hepatitis treatment; significant side effects, with over 80% of patients experiencing flu-like symptoms such as fever and myalgia; long-term use may also lead to myelosuppression, thyroid dysfunction, and neuropsychiatric problems such as depression; its applicable population is limited, and it is contraindicated in patients with decompensated cirrhosis or active autoimmune diseases; the administration method is subcutaneous or intramuscular injection, and the treatment course is as long as 6-12 months, resulting in poor patient compliance.

[0003] Interferon-alpha, as an antiviral drug, has several significant drawbacks in clinical application. Its efficacy is limited; the antiviral response rate is greatly affected by viral genotype, baseline viral load, and the patient's immune status. For example, in the treatment of chronic hepatitis B and C, the overall response rate is only 30%-50%, and some patients with certain viral genotypes may not even achieve ideal results. Side effects are frequent and involve multiple systems. Over 80% of patients experience flu-like symptoms such as fever, fatigue, and myalgia at the beginning of treatment. Long-term use can easily lead to bone marrow suppression, resulting in leukopenia and thrombocytopenia, and may also induce thyroid dysfunction, exacerbate autoimmune diseases, and cause neuropsychiatric symptoms such as depression and insomnia in some patients, requiring discontinuation of the drug in severe cases. The applicable population is narrow; it is contraindicated in patients with decompensated cirrhosis, severe cardiopulmonary insufficiency, active autoimmune diseases, and pregnancy, preventing many patients from using it due to contraindications. In addition, the medication is inconvenient to administer, requiring subcutaneous or intramuscular injection, and the treatment course lasts for 6-12 months. The cumbersome medication method and long treatment course significantly reduce patient compliance, thereby affecting the treatment effect.

[0004] This invention confirms that lactate produced by the host in the later stages of viral infection is a key factor leading to the ineffectiveness of interferon-alpha or the occurrence of severe side effects: lactate not only directly inhibits the antiviral activity of interferon-alpha, but can also induce a cytokine storm in the presence of interferon-alpha, exacerbating host damage and doubly weakening the clinical efficacy of interferon-alpha. Based on this, we developed a specific combination therapy—interferon combined with the lactate dehydrogenase inhibitor stiripentol (STP, an FDA-approved antiepileptic drug for clinical use). By reducing host lactate release, this therapy reverses the lactate-mediated "antiviral-pro-inflammatory" effect, achieving the dual benefits of interferon in "antiviral and anti-inflammatory control." This combination therapy can effectively overcome the clinical limitations of interferon, improve the treatment effect in the late stages of infection, and avoid side effects such as cytokine storms. It provides a new strategy for the development of broad-spectrum antiviral drugs and also offers new drug options and treatment directions for the prevention and control of existing epidemic viruses and the response to pandemics of unknown viruses. Summary of the Invention

[0005] The present invention first provides an antiviral drug composition comprising interferon α and stearyl alcohol.

[0006] In some embodiments, the weight ratio of interferon α to stearyl alcohol is 1:10 to 10:1.

[0007] In some embodiments, the concentration of the stearyl alcohol used is from 10 μM to 100 μM.

[0008] In some embodiments, the concentration of interferon α used is from 0.1 nM to 10 nM.

[0009] In some embodiments, the antiviral drug composition is used to inhibit viral replication, including influenza A virus (H1N1) or vesicular stomatitis virus (VSV).

[0010] In some embodiments, the antiviral pharmaceutical composition is used to reduce the expression of inflammatory factors induced by viral infection, including IL-1α, IL-6, or TNF-α.

[0011] The present invention also provides the use of interferon-α and staefenol in the preparation of antiviral drugs, said drugs being used in combination to enhance antiviral effects and reduce side effects.

[0012] In some embodiments, the combined administration method is simultaneous or sequential administration.

[0013] In some embodiments, the antiviral drug is applicable to mammals, including humans.

[0014] In some embodiments, the mammal is a human.

[0015] Compared with the prior art, the present invention has at least the following beneficial effects: (1) Enhance antiviral efficacy: The combination therapy can reverse lactate-mediated inhibition of interferon α activity and significantly enhance the ability to inhibit viral replication, especially showing broad-spectrum effects against influenza A virus and vesicular stomatitis virus.

[0016] (2) Reduce side effects: Staphylol reduces lactic acid accumulation, effectively avoids inflammatory reactions such as cytokine storms, and reduces flu-like symptoms such as fever and myalgia, as well as the risk of bone marrow suppression caused by long-term use.

[0017] (3) Expand the applicable population: This combination regimen has high safety and can be potentially used for patients with original contraindications such as decompensated cirrhosis, thus improving clinical adaptability.

[0018] (4) Improve medication convenience: Combination therapy may shorten the course of treatment or optimize the administration method, thereby enhancing patient compliance.

[0019] (5) Overall, this invention provides an innovative direction for antiviral treatment, combining high efficiency and safety. Attached Figure Description

[0020] Figure 1 This graph shows the dynamic changes in lactate content after viral infection of cells. H1N1: Influenza A virus (H1N1); VSV: Vesicular Stomatitis Virus; LAC: Lactic acid (10 mM); *** indicates p < 0.0001, indicating a statistically significant difference; **** indicates p < 0.00001, indicating a statistically significant difference.

[0021] Figure 2 This diagram illustrates the pro-inflammatory effects of lactic acid during viral infection. Figure 2 In the figure, A represents the result of VSV infection of mouse macrophages with IL-1α; Figure 2 B in the figure represents the IL-6 result of VSV infection of mouse macrophages; Figure 2 In the figure, C represents the TNF-α result of VSV infection of mouse macrophages; Figure 2 In the figure, D represents the result of IL-1α infection of mouse macrophages by H1N1; Figure 2 E in the figure represents the IL-6 result of H1N1 infection of mouse macrophages; Figure 2In the table, F represents the TNF-α result of H1N1 infection of mouse macrophages; H1N1: influenza A virus H1N1; VSV: vesicular stomatitis virus; LAC: lactic acid (10mM); ns: p>0.05, no significant difference; ** indicates p<0.001, statistically significant difference; *** indicates p<0.0001, statistically significant difference; **** indicates p<0.00001, statistically significant difference.

[0022] Figure 3 The diagram shows the inhibitory effect of the combination of stapfenol and interferon-α on inflammation. Figure 3 In the figure, A represents the result of VSV infection of mouse macrophages with IL-1α; Figure 3 B in the figure represents the IL-6 result of VSV infection of mouse macrophages; Figure 3 In the figure, C represents the TNF-α result of VSV infection of mouse macrophages; Figure 3 In the figure, D represents the result of IL-1α infection of mouse macrophages by H1N1; Figure 3 E in the figure represents the IL-6 result of H1N1 infection of mouse macrophages; Figure 3 In the table, F represents the TNF-α result of H1N1 infection of mouse macrophages; H1N1: influenza A virus H1N1; VSV: vesicular stomatitis virus; STP: lactate dehydrogenase inhibitor stiripentol (STP); * indicates p<0.05, indicating a statistically significant difference; ** indicates p<0.001, indicating a statistically significant difference; *** indicates p<0.0001, indicating a statistically significant difference; **** indicates p<0.00001, indicating a statistically significant difference.

[0023] Figure 4 The combined use of STP and IFN α can significantly inhibit viral replication. Figure 4 In this context, A represents the result of VSV infection of MLF cells; Figure 4 B in the figure represents the result of H1N1 infection of MLF cells; Figure 3In the table, C represents the TNF-α result of VSV infection of mouse macrophages; H1N1: influenza A virus H1N1; VSV: vesicular stomatitis virus; STP: lactate dehydrogenase inhibitor stiripentol (STP); * indicates p<0.05, indicating a statistically significant difference; ** indicates p<0.001, indicating a statistically significant difference; *** indicates p<0.0001, indicating a statistically significant difference; **** indicates p<0.00001, indicating a statistically significant difference. Detailed Implementation

[0024] To make the technical problems, technical solutions and advantages of the present invention clearer, a detailed description will be given below in conjunction with the accompanying drawings and specific embodiments.

[0025] Stiripentol (STP), CAS No.: 49763-96-4, purchased from Yuan Ye, product number: S85554.

[0026] Example 1: Large amounts of lactic acid are produced when cells are infected with a virus. We infected A549 cells with influenza A virus (H1N1) and vesicular stomatitis virus (VSV). There were three experimental groups: a control group (MOCK), virus-infected group 1 (H1N1), and virus-infected group 2 (VSV). The virus infection concentration was 1.0 MOI, and the lactate concentration was 10 nM. The lactate content in the cell supernatant was measured at 6, 12, 24, and 48 hours after virus infection. The results are shown below. Figure 1 As shown, it was found that after the virus infected the cells, it caused the cells to produce a large amount of lactic acid.

[0027] Example 2: Detection of the pro-inflammatory effect of lactic acid in cells infected with viruses Mouse peritoneal macrophages were extracted and divided into three experimental groups: a control group (MOCK, a simulated treatment group, used as an experimental control without virus inoculation, only supplemented with the same dose of virus preservation solution), a virus infection group (H1N1 or VSV), and a virus infection plus lactate stimulation group (H1N1+LAC or VSV+LAC). The virus infection concentration was 1.0 MOI, and the lactate concentration was 10 nM. After 24 hours, cells were collected, total RNA was extracted, and the changes in the expression levels of inflammatory factors IL-1α, IL-6, and TNF-α in cells were analyzed by quantitative real-time PCR (QRT-PCR). Figure 2 The results showed that viral infection combined with lactic acid stimulation exacerbated the expression levels of intracellular inflammatory factors.

[0028] Example 3: The combined use of the lactate dehydrogenase inhibitor stiripentol (STP) and IFN-α was found to inhibit inflammation caused by viral infection. Mouse peritoneal macrophages were extracted and divided into three experimental groups: a control group (MOCK, a simulated treatment group, used as an experimental control without virus inoculation, only adding the same dose of virus preservation solution), a virus infection group (H1N1 or VSV), and a virus infection plus drug treatment group (H1N1+STP+IFNα or VSV+STP+IFNα). The virus infection concentration was 1.0 MOI, the STP concentration was 50 μM, and the IFNα concentration was 2 nM. After 24 hours, cells were collected, total RNA was extracted, and the changes in the expression levels of inflammatory factors IL-1α, IL-6, and TNF-α in cells were analyzed by real-time quantitative PCR (QRT-PCR). Figure 3 The results showed that STP combined with IFNα treatment after viral infection could inhibit the expression of intracellular inflammatory factors.

[0029] Example 4: The combined use of the lactate dehydrogenase inhibitor stiripentol and IFN-α was detected to inhibit viral replication. Mouse lung fibroblasts (MLF) were used and divided into four experimental groups: a control group (MOCK, a simulated treatment group, serving as an experimental control without virus inoculation, only supplemented with the same dose of virus preservation solution), a virus infection group (H1N1 or VSV), a virus infection plus drug treatment group 1 (H1N1+IFNα or VSV+IFNα), and a virus infection plus drug treatment group 2 (H1N1+STP+IFNα or VSV+STP+IFNα). The virus infection concentration was 1.0 MOI, the STP concentration was 50 μM, and the IFNα concentration was 2 nM. After 24 hours, cells were collected, total RNA was extracted, and changes in viral replication levels in cells were analyzed by real-time quantitative PCR (QRT-PCR). The results showed that IFNα treatment alone could inhibit viral replication to some extent, but the combination of IFNα and STP significantly inhibited viral replication.

[0030] The above description represents the preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. An antiviral drug composition, characterized in that, It contains interferon-alpha and stearyl alcohol.

2. The antiviral pharmaceutical composition according to claim 1, characterized in that, The weight ratio of interferon α to stearyl alcohol is 1:10 to 10:

1.

3. The antiviral pharmaceutical composition according to claim 1, characterized in that, The concentration of the stearyl alcohol used is from 10 μM to 100 μM.

4. The antiviral pharmaceutical composition according to claim 1, characterized in that, The concentration of interferon α used is from 0.1 nM to 10 nM.

5. The antiviral pharmaceutical composition according to claim 1, characterized in that, The antiviral drug composition is used to inhibit viral replication, including influenza A virus (H1N1) or vesicular stomatitis virus (VSV).

6. The antiviral pharmaceutical composition according to claim 1, characterized in that, The antiviral drug composition is used to reduce the expression of inflammatory factors induced by viral infection, including IL-1α, IL-6, or TNF-α.

7. The use of interferon α and staepentol in the preparation of antiviral drugs, characterized in that, The drugs are used in combination to enhance antiviral effects and reduce side effects.

8. The use as described in claim 7, characterized in that, The combined administration method is simultaneous or sequential administration.

9. The use as described in claim 7, characterized in that, The antiviral drugs are applicable to mammals, including humans.

10. The use as described in claim 9, characterized in that, The mammal in question is a human.

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