Application of BGT226 maleate in preparation of medicine for resisting novel coronavirus

Abstract

The invention discloses application of BGT226 maleate in preparation of novel coronavirus resisting drugs, and belongs to the technical field of antiviral drugs. According to the application of the BGT226 maleate in preparation of the novel coronavirus resisting medicine, tests find that the IC50 of the BGT226 maleate for an SARS-CoV-2 original strain is 0.116 mu M, the IC50 of the BGT226 maleate for an Omicro variant (JN.1) is 0.084 mu M, and the BGT226 maleate has good biological safety, can be used for research and development of the novel coronavirus infection resisting medicine, is a potential medicine for treating novel coronavirus infection, and has wide application prospects. And a new direction and possibility are provided for the treatment of the novel coronavirus and the variant thereof.

Description

TECHNICAL FIELD

[0001] The application relates to the field of antiviral drugs, in particular to application of BGT226 maleate in preparation of an anti-2019-nCoV drug. BACKGROUND

[0002] At present, the research and development strategies of anti-2019-nCoV drugs mainly include target drug design and development aiming at key virus molecules (such as Mpro, RdRP, PLpro, etc.) and host factors (such as ACE2, TMPRSS2, etc.), and drug screening based on a virus infection model. The former develops small molecule inhibitors, polypeptides, aptamers, neutralizing antibodies, etc. through virtual screening or experimental screening based on molecular interaction, and develops neutralizing antibodies through antigen immune screening or antibody gene sequencing. The latter screens drugs or compounds that can inhibit virus replication by testing various existing drugs and compound libraries (including known active compound libraries, natural product compound libraries, etc.).

[0003] BGT226 maleate is a maleate modified form of BGT226, and the modification can improve the water solubility of the drug molecule. BGT226 is a PI3K / mTOR dual inhibitor developed by Novartis, with IC50 of 4 nM, 63 nM and 38 nM for PI3Kα, PI3Kβ and PI3Kγ, respectively. Since the PI3K / Akt / mTOR pathway plays an important role in cell proliferation and survival, inhibition of the pathway causes cell cycle arrest and apoptosis, thus producing a tumor cell-killing effect. Studies have shown that BGT226 has strong growth inhibitory activity on various tumor cells. The results of clinical phase I trials show that it has good pharmacokinetics and dose range. However, the therapeutic effect of BGT226 maleate in 2019-nCoV infection has not been studied. SUMMARY

[0004] The purpose of the present application is to provide the application of BGT226 maleate in the preparation of an anti-2019-nCoV drug, which is a new application of BGT226 maleate in 2019-nCoV infection and has good development prospects.

[0005] To achieve the above purpose, the application provides the application of BGT226 maleate in the preparation of an anti-2019-nCoV drug, the molecular formula of BGT226 maleate is C 32 H 29 F3N6O6, and the structural formula is as follows: .

[0006] Preferably, the 2019-nCoV includes SARS-CoV-2 original strain and Omicron variant JN.1.

[0007] Preferably, the IC50 of BGT226 maleate against the original strain of SARS-CoV-2 is 0.116 μM.

[0008] Preferably, the IC50 of BGT226 maleate against the Omicron variant (JN.1) is 0.084 μM.

[0009] Therefore, the application of BGT226 maleate in the preparation of anti-novel coronavirus drugs, the test found that the IC50 of BGT226 maleate against the original strain of SARS-CoV-2 is 0.116 μM, and the IC50 of BGT226 maleate against the Omicron variant (JN.1) is 0.084 μM, and it has good biological safety, which reveals that it can be used for the development of anti-novel coronavirus infection drugs, and is a potential drug for the treatment of novel coronavirus infection, and provides a new direction and possibility for the treatment of novel coronavirus and its variants.

[0010] The technical solutions of the present application will be further described below with the aid of the accompanying drawings and examples. BRIEF DESCRIPTION OF DRAWINGS

[0011] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings needed to be used in the description of the embodiments of the present application will be briefly introduced. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can also be obtained by those skilled in the art without creative labor.

[0012] Figure 1 The graph of virus inhibition percentage for different treatment groups; Figure 2 The graph of cell morphology observation for different treatment groups. DETAILED DESCRIPTION

[0013] The technical solutions of the present application will be further described below with the aid of the accompanying drawings and examples.

[0014] In order to make the purpose, technical solutions and advantages of the present application more clear, thorough and complete, the technical solutions of the present application will be described clearly and completely below with the aid of the drawings and examples. The following detailed description is a description of the embodiments, which aims to provide further detailed description of the present application. Unless otherwise specified, all technical terms used in the present application have the same meaning as generally understood by those skilled in the art to which the present application belongs.

[0015] The instruments and reagents used in the examples were obtained through commercial channels.

[0016] The molecular formula of BGT226 maleate used is: C 32 H29 F3N6O6, the structural formula of which is: .

[0017] Examples Constructing a new coronavirus drug screening model: (1) African green monkey kidney Vero cells were inoculated in DMEM medium and placed in a 37°C, 5% CO2 incubator for culture. One day before drug screening, 2.5 x 10 4 A number of Vero cells were inoculated in a 96-well plate so that the density reached more than 90% the next day. The initial concentration of the small molecule kinase inhibitor library was 10 mM, which was first diluted to 100 μM in DMEM medium, and then diluted to 7 concentrations with a 2-fold gradient. 8 μL of the diluted drug was aspirated and added to the Vero cells in the culture plate, with a medium content of 150 μL per well.

[0018] (2) Infection with the original strain and Omicron variant (JN.1) of SARS-CoV-2 was performed in a biosafety level 3 laboratory. The original virus liquid was diluted with DMEM medium at an appropriate ratio, and 50 μL of the diluted virus liquid was added to the culture well so that the final infection titer reached 100 times TCID50, and the final drug concentration reached 4, 2, 1, 0.5, 0.25, 0.125, 0.0625 and 0.03125 μM, respectively. At the same time, a virus-free infection negative control and a virus addition without drug positive control were set up, each with 3 replicates.

[0019] (3) After 3 days of viral infection, the viral load in the culture supernatant was detected using a new coronavirus RNA detection kit (Shanghai Biomerieux Medical Technology Co., Ltd., Catalog No.: ZC-HX-201-02, National Medical Equipment Accreditation 20203400065) to evaluate the inhibitory effect of the drug on viral replication, and the cell pathology was observed under a microscope. If high-concentration drugs are found to cause cell pathology and death, the drug concentration is considered to be ineffective.

[0020] The test procedure is as follows: After culture, 100 µL of culture supernatant was collected and viral nucleic acid was extracted using a magnetic bead nucleic acid extraction kit and an automated nucleic acid extractor (Shanghai ZJ Biotechnology). The viral RNA level was detected using a novel coronavirus nucleic acid detection kit (fluorescent PCR method): Specific primers and probes were designed for the ORF1a / b and N genes of the novel coronavirus RNA (NC_045512.2) using a one-step RT-PCR method combined with TaqMan technology. The ORF1a / b gene probe was labeled with FAM fluorescein, the N gene probe with VIC fluorescein, and the internal control probe with ROX fluorescein. After 45 cycles of PCR denaturation (97℃, 5s), annealing, and extension (58℃, 30s), the light signals emitted after hydrolysis of the probes labeled with different fluoresceins were collected by the PCR instrument, displaying amplification curves on the instrument, thus enabling the detection of the novel coronavirus ORF1a / b and N genes.

[0021] The level of the virus is represented by the Ct value, through 2 -ΔΔCt The method calculates the relative amount of viral nucleic acid in each drug-treated group and the drug-free group to obtain the percentage of viral inhibition, and evaluates the drug's activity against the novel coronavirus.

[0022] Virus inhibition percentage such as Figure 1 As shown, the microscopic images of the cells are as follows: Figure 2 As shown, 2, 1, 0.5, 0.25, 0.125, 0.0625, and 0.03125 μM BGT226 maleate can inhibit viral replication without significantly altering cell morphology. Although 4 μM BGT226 maleate inhibited viral replication, it significantly altered cell morphology, indicating that this drug concentration caused considerable cytotoxicity and therefore was not included in the effective concentration.

[0023] In summary, the IC50 of BGT226 maleate against the original SARS-CoV-2 strain is 0.116 μM, and the IC50 against the Omicron variant (JN.1) is 0.084 μM.

[0024] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. The application of BGT226 maleate in the preparation of drugs against the novel coronavirus, characterized in that, BGT226 maleate includes its pharmaceutically acceptable salts and derivatives. The molecular formula of BGT226 maleate is C226. 32 H 29 F3N6O6, the structural formula is shown below: 。 2. The application of BGT226 maleate according to claim 1 in the preparation of anti-novel coronavirus drugs, characterized in that: The novel coronavirus includes the original SARS-CoV-2 strain and the Omicron variant JN.

1.

3. The application of BGT226 maleate according to claim 2 in the preparation of anti-novel coronavirus drugs, characterized in that: The IC50 of BGT226 maleate against the original SARS-CoV-2 strain is 0.116 μM.

4. The application of BGT226 maleate according to claim 2 in the preparation of anti-novel coronavirus drugs, characterized in that: The IC50 of BGT226 maleate against the Omicron mutant JN.1 is 0.084 μM.

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