Application of Voxtalisib in the preparation of anti-ADV7 virus drugs

Abstract

This patent discloses the use of voxtalisib in the preparation of anti-ADV7 drugs. Experimental studies of voxtalisib's anti-ADV7 activity have shown that the compound voxtalisib inhibits the cytopathic effect (CPE) produced by ADV7 in Hela cells, enhancing cell survival and reducing progeny virus production, suggesting its potential application in the preparation of anti-ADV7 drugs.

Description

Technical Field

[0001] The present invention relates to the technical field of antiviral drugs and relates to the use of Voxtalisib in the preparation of anti-ADV7 virus drugs. Background Art

[0002] Human adenovirus type 7 (ADV7) is a widespread, non-enveloped, double-stranded DNA virus with a diameter of 90-100 nm and a dihedral capsid. Its replication is independent of host cell division. Human adenovirus (ADV) infections are common and can cause a variety of diseases, primarily respiratory tract infections. Three types of adenovirus are primarily associated with respiratory diseases: ADV-3, ADV-5, and ADV-7. Risk factors for severe disease are known to be relatively low. Commonly, it can cause acute respiratory fever, but it is particularly prevalent in children, with severe cases potentially leading to pneumonia in children. ADV7 has been shown to be a significant cause of pneumonia in children. ADV7 can cause severe or fatal respiratory tract infections in children under seven years of age, and can easily lead to outbreaks, posing a health threat to infants and young children. Currently, there is a lack of clinically specific drugs with minimal side effects against ADV7 infection, and symptomatic treatment is currently the only approach. Therefore, the development of specific anti-ADV7 drugs is imperative.

[0003] Voxtalisib is the first orally available dual inhibitor of PI3K and mTOR, with IC50 values ​​of 39, 113, 9, 43, and ~150 nM for p110α, β, γ, δ, and mTOR, respectively. In vitro, voxtalisib reduces cell viability in a concentration-dependent manner. Oral administration of voxtalisib to mice resulted in a 12-fold reduction in the number of metastatic tumors compared to the control group. Voxtalisib has demonstrated activity both alone and in combination with other agents against various GBM xenografts. Voxtalisib downregulates the expression of PI3K downstream phosphoproteins pAKT and pPRAS40, as well as mTOR downstream phosphoproteins pS6 and p4EBP1, in cultured human xenografts. However, the use of voxtalisib for antiviral therapy has been rarely reported.

[0004] We evaluated the inhibitory activity of voxtalisib against ADV7 virus. Summary of the Invention

[0005] The purpose of the present invention is to provide Voxtalisib as an effective inhibitor of ADV7 virus in response to the above situation.

[0006] The purpose of the present invention is achieved by testing the activity of Voxtalisib using a standard viral activity test method.

[0007] Through extensive biological experiments, the applicant has discovered that voxtalisib exhibits anti-ADV7 activity. Specifically, it can inhibit the cytopathic effect caused by ADV7, enhance the survival rate of infected cells, inhibit ADV7 replication and proliferation within cells, and reduce progeny virus production. This suggests that this compound has the potential to be used as a specific therapeutic against ADV7 infection and has great clinical application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0008] Figure 1 The effect of Voxtalisib on the survival rate of Hela cells treated with ADV7.

[0009] Figure 2 It is the inhibitory effect of Voxtalisib on CPE of Hela cells caused by ADV7.

[0010] Figure 3 It is the inhibitory effect of Voxtalisib on the production of ADV7 progeny virus. DETAILED DESCRIPTION

[0011] Voxtalisib used in the present invention was purchased from a reagent company.

[0012] The present invention relates to the application of Voxtalisib in the preparation of anti-ADV7 virus drugs.

[0013] The application refers to the addition of pharmaceutically acceptable excipients and carriers to Voxtalisib for the preparation of anti-ADV7 virus preparations.

[0014] The preparation is granules, tablets, pills, capsules, injections or dispersions.

[0015] The present invention uses voxtalisib as an anti-ADV7 virus, and found that voxtalisib has a significant anti-ADV7 virus effect, which shows that this compound has the potential to prepare a specific therapeutic drug for anti-ADV7 infection and has great clinical application prospects.

[0016] The applicant has conducted an anti-ADV7 activity study experiment on the above compounds, and the experimental results are as follows: In the following, unless otherwise specified, the materials and operating methods used in the present invention are well known in the art.

[0017] 1. Test content:

[0018] Analysis of compound anti-ADV7 activity: The present invention combines cytopathic effect analysis and MTT cell viability assay to evaluate the anti-ADV7 activity of Voxtalisib.

[0019] 2. Test method:

[0020] 2.1.1 Toxicity of the compounds to host Hela cells

[0021] Hele cells were plated in 96-well plates and cultured in a 37°C, 5% CO2 incubator until a confluent monolayer was formed. The cell culture medium was discarded and cell maintenance medium containing different concentrations of the test compound was added and cultured continuously. After 48 hours, the cytotoxicity was observed under a microscope and recorded. The cell survival rate was determined by MTT assay. The median cytotoxic concentration (CC) of the drug for the cells was calculated using SPSS 11.5 software. 50 ). Cell viability = (mean OD of drug group 492 Value / average OD of cell control group 492 value) × 100%.

[0022] 2.1.2 Inhibitory activity of compounds against ADV7

[0023] HeLa cells were plated in 96-well plates and cultured in a 37°C, 5% CO2 incubator until confluent monolayers were achieved. The culture medium was then discarded and the cells were infected with 100 TCID50 of ADV7 virus for 1.5 hours. The cells were then incubated with varying concentrations of the test compound (ribavirin was used as a positive control). After approximately 48 hours of incubation, when approximately 90% of the virus control wells showed CPE, the cells were observed microscopically. CPE was recorded as follows: no cytopathic effect (CPE) was scored as -, less than 25% cytopathic effect (CPE) as +, 25%-50% cytopathic effect (CPE) as ++, 50%-75% cytopathic effect (CPE) as +++, and greater than 75% cytopathic effect (CPE) as ++++.

[0024] After the CPE observation was completed, the MTT method was used to detect the inhibitory rate of the drug on ADV7. The specific steps were as follows: 50 μL of MTT (5 mg mL -1 After incubation for 3-4 h, the supernatant was removed and an equal volume of DMSO was added to dissolve the precipitate. The corresponding absorbance (OD) was read at 492 nm using a microplate reader. 492 The inhibition rate of the drug on ADV7 was calculated using the following formula. The concentration for 50% of maximal effect (EC) was calculated using SPSS 11.5 software. 50 ).

[0025]

[0026] 2.1.3 Therapeutic Index (TI) of Drugs

[0027] TI = CC 50 / EC 50 The higher the therapeutic index, the greater the antiviral potential.

[0028] 3. Experimental results

[0029] Table 1 Voxtalisib cytotoxicity and anti-ADV7 activity

[0030]

[0031] The results of the compound cytotoxicity and anti-ADV7 activity tests are shown in Table 1. The concentration-dependent effects of the compounds on the survival rate of Hela cells treated with ADV7 are shown in Table 1. Figure 1 It was found that at 25µM, the maximum inhibition rate of Voxtalisib was approximately 91.1%.

[0032] The applicant has conducted in-depth research on the anti-ADV7 activity of Voxtalisib and conducted a test on the compound's inhibitory effect on the production of ADV7 progeny viruses. The test results are as follows:

[0033] 1. Test content

[0034] The inhibitory effect of the compound on the production of ADV7 progeny virus was detected after ADV7 infected Hela cells.

[0035] 2. Test methods

[0036] HeLa cells in the logarithmic growth phase were plated in 24-well plates. After the cells grew into a confluent monolayer, 100 TCID 50 ADV7 infected cells, incubated at 37℃ for 1.5 h, removed the virus solution, washed three times with PBS, and added cell maintenance medium containing 6.25μM concentration. After 48 h, cells and supernatant culture medium were collected and lysed three times at -20℃ and 37℃, and TCID 50 Methods The ADV7 virus titer was determined.

[0037] 3. Test results

[0038] like Figure 2 As shown, the cell survival rate of Hela cells infected with ADV7 treated with Voxtalisib was significantly increased compared with the virus control group, indicating that the compound can significantly inhibit the activity of ADV7.

[0039] like Figure 3 As shown, the virus titer of Hela cells treated with the compound was significantly decreased compared with the virus control group, which was decreased by 4.24 log compared with the virus control.

[0040] In summary, Voxtalisib has a strong inhibitory activity against ADV7, can inhibit Hela cell death caused by ADV7 virus, and can be prepared into a clinically effective drug against ADV7 infection.

[0041] It should be understood that the above embodiments are only intended to illustrate the present invention and are not intended to limit the scope of protection of the present invention. In addition, it should be understood that after reading the contents of the present invention, those skilled in the art may make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

Claims

1. Application of Voxtalisib in the preparation of anti-ADV7 virus drugs.

2. The use of Voxtalisib in the preparation of anti-ADV7 virus drugs according to claim 1, characterized in that: Voxtalisib inhibited ADV7 by approximately 91.1% at 25µM.

3. The use of Voxtalisib in the preparation of anti-ADV7 virus drugs according to claim 1, characterized in that: The application refers to adding pharmaceutically acceptable excipients to this compound to prepare an anti-ADV7 virus preparation.

4. The use of Voxtalisib in the preparation of anti-ADV7 virus drugs according to claim 3, characterized in that: The preparation is in the form of granules, tablets, pills, capsules, injections or dispersions.

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