Use of bictegravir for the preparation of a medicament and medicaments

Abstract

The application discloses application of Bictegravir in preparation of a medicine and the medicine. In the application of Bictegravir in preparation of the medicine, the medicine is used for inhibiting Mycobacterium tuberculosis. It is found for the first time that Bictegravir has the effect of inhibiting intracellular survival of Mycobacterium tuberculosis at a human macrophage level, in the application, in the application of Bictegravir in preparation of the medicine, the medicine can effectively inhibit Mycobacterium tuberculosis, Bictegravir can be used for preparing a medicine for preventing and / or treating tuberculosis, and Bictegravir can be used for preparing a medicine for preventing and / or treating tuberculosis and AIDS.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to the application of Bictegravir in the preparation of drugs and the drugs themselves. Background Technology

[0002] Acquired immunodeficiency syndrome (AIDS) is a significant public health problem threatening public health. AIDS patients are immunocompromised and often have multiple opportunistic infections, among which Mycobacterium tuberculosis (MTB) is a common pathogen, and the disease is often progressive. Individuals with HIV infection and latent Mycobacterium tuberculosis infection (LTBI) are 30 times more likely to progress to active tuberculosis than HIV-negative individuals. Therefore, HIV and MTB interact and influence each other, making diagnosis and treatment complex and challenging. Summary of the Invention

[0003] The main objective of this invention is to propose an application of Bictegravir in the preparation of a drug and a drug that can effectively fight HIV while also effectively inhibiting Mycobacterium tuberculosis.

[0004] To achieve the above objectives, in a first aspect, the present invention proposes the use of Bictegravir in the preparation of a drug for inhibiting Mycobacterium tuberculosis.

[0005] Optionally, the drug is used to inhibit Mycobacterium tuberculosis H37Rv and / or H37Ra.

[0006] Secondly, the present invention proposes the use of Bictegravir in the preparation of a drug for the prevention and / or treatment of a disease, or for reducing the risk of a disease, wherein the disease is tuberculosis.

[0007] Thirdly, the present invention proposes the use of Bictegravir in the preparation of a drug for the prevention and / or treatment of a disease, or for reducing the risk of a disease; the disease being tuberculosis and AIDS.

[0008] Fourthly, the present invention provides a drug for inhibiting Mycobacterium tuberculosis, the drug comprising Bictegravir.

[0009] Fifthly, the present invention provides a medicament for the prevention and / or treatment of tuberculosis, the medicament comprising Bictegravir.

[0010] Optionally, the drug may further comprise pharmaceutically acceptable excipients, carriers, and / or diluents.

[0011] In the technical solution of this invention, it is discovered for the first time that Bictegravir has the effect of inhibiting the intracellular survival of Mycobacterium tuberculosis at the human macrophage level. In the application of Bictegravir in the preparation of drugs, the drug can effectively inhibit Mycobacterium tuberculosis. Furthermore, Bictegravir can be used to prepare drugs for the prevention and / or treatment of tuberculosis. Furthermore, Bictegravir can be used to prepare drugs for the prevention and / or treatment of tuberculosis and AIDS. Attached Figure Description

[0012] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the structures shown in these drawings without creative effort.

[0013] Figure 1 This is a graph showing the cell viability of THP-1 cells treated with different concentrations of Bictegravir in Example 1 of this invention.

[0014] Figure 2 This describes the inhibitory effect of 10 μM Bictegravir on Mycobacterium tuberculosis H37Rv at the THP-1 macrophage level in Example 2 of this invention.

[0015] Figure 3 This is an example of the inhibitory effect of 10 μM Bictegravir on Mycobacterium tuberculosis H37Ra at the THP-1 macrophage level in Example 3 of the present invention.

[0016] The realization of the objective, functional features and advantages of the present invention will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0017] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Where the manufacturers of reagents or instruments are not specified, they are all conventional products that can be purchased commercially. Furthermore, the meaning of "and / or" throughout the text includes three parallel solutions; for example, "A and / or B" includes solution A, or solution B, or a solution where both A and B are satisfied simultaneously. In addition, the technical solutions of the various embodiments can be combined with each other, but this must be based on the ability of those skilled in the art to implement them. When the combination of technical solutions is contradictory or cannot be implemented, it should be considered that such a combination of technical solutions does not exist and is not within the scope of protection claimed by the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0018] Acquired immunodeficiency syndrome (AIDS) is a significant public health problem threatening public health. AIDS patients are immunocompromised and often have multiple opportunistic infections, among which Mycobacterium tuberculosis (MTB) is a common pathogen, and the disease is often progressive. Individuals with HIV infection and latent Mycobacterium tuberculosis infection (LTBI) are 30 times more likely to progress to active tuberculosis than HIV-negative individuals. Therefore, HIV and MTB interact and influence each other, making diagnosis and treatment complex and challenging.

[0019] In view of this, embodiments of the present invention propose an application of Bictegravir in the preparation of a drug and a drug thereof.

[0020] In a first aspect, embodiments of the present invention propose the application of Bictegravir in the preparation of a drug for inhibiting Mycobacterium tuberculosis.

[0021] Bictegravir (BIC) is a second-generation integrase strand transfer inhibitor (INSTI) used in combination with emtricitabine and tenofovir alafenamide at a fixed dose for the treatment of HIV. It exhibits potent antiviral activity in vitro against wild-type HIV and strains resistant to first-generation integrase strand transfer inhibitors. This invention is the first to discover that Bictegravir inhibits the intracellular survival of Mycobacterium tuberculosis at the human macrophage level.

[0022] In some embodiments, the drug is used to inhibit Mycobacterium tuberculosis H37Rv and / or H37Ra.

[0023] Secondly, the present invention proposes the use of Bictegravir in the preparation of a drug for the prevention and / or treatment of a disease, or for reducing the risk of a disease, wherein the disease is tuberculosis.

[0024] In this invention, the term "prevention and / or treatment" includes not only the prevention and / or treatment of disease, but also generally includes reducing or inhibiting the number of pathogens associated with the disease, preventing the onset of the disease, slowing or reversing the progression of the disease, preventing or slowing the onset of one or more symptoms associated with the disease, reducing and / or alleviating one or more symptoms associated with the disease, reducing the severity and / or duration of the disease and / or any symptoms associated with it and / or preventing a further increase in the severity of the disease and / or any symptoms associated with it, preventing, reducing or reversing any physiological damage caused by the disease, and any pharmacological effects that are generally beneficial to the patient being treated. The pharmaceuticals of this application do not need to achieve a complete cure or eradication of any symptoms or manifestations of the disease to be considered useful therapeutic agents. As recognized in the relevant art, pharmaceuticals used as therapeutic agents may reduce the severity of a given disease state, but do not need to eliminate every manifestation of the disease to be considered useful therapeutic agents. Similarly, preventatively applied treatments constituting feasible preventative agents do not need to completely and effectively prevent the onset of the disease. Simply reducing the impact of the disease on the subject (e.g., by reducing the number or severity of their symptoms, or by increasing the effectiveness of another treatment, or by producing another beneficial effect), or reducing the likelihood of the disease occurring or worsening, is sufficient.

[0025] Thirdly, the present invention proposes the use of Bictegravir in the preparation of a drug for the prevention and / or treatment of a disease, or for reducing the risk of a disease; the disease being tuberculosis and AIDS.

[0026] Bictegravir, when used in combination with emtricitabine and tenofovir alafenamide at a fixed dose to treat HIV, exhibits potent antiviral activity in vitro against wild-type HIV and strains resistant to first-generation integrase strand transfer inhibitors. This embodiment describes the application of Bictegravir in the preparation of a drug for the prevention and / or treatment of diseases, or for reducing the risk of diseases such as tuberculosis and AIDS. This achieves effective anti-HIV action while simultaneously inhibiting Mycobacterium tuberculosis.

[0027] Fourthly, the present invention provides a drug for inhibiting Mycobacterium tuberculosis, the drug comprising Bictegravir.

[0028] Fifthly, the present invention provides a medicament for the prevention and / or treatment of tuberculosis, the medicament comprising Bictegravir.

[0029] In some embodiments, the drug further comprises pharmaceutically acceptable excipients, carriers, and / or diluents.

[0030] The technical solution of the present invention will be further described in detail below with reference to specific embodiments and accompanying drawings. It should be understood that the following embodiments are only used to explain the present invention and are not intended to limit the present invention.

[0031] Example 1:

[0032] Differentiation of THP-1 macrophages: THP-1 cells were purchased from the Cell Bank of the Chinese Academy of Sciences and cultured in 1640 medium containing 10% fetal bovine serum at 37°C in a cell culture incubator with 5% CO2. The cells were seeded in 96-well plates at a density of 2.5 × 10⁴ cells per well and stimulated overnight with a final concentration of 50 ng / ml of PMA to differentiate into macrophages. After 24 hours, the medium was replaced with complete medium.

[0033] Cell viability determination: After culturing the induced macrophages for another 24 h, Bictegravir was added at concentrations of 5 μM, 10 μM, 20 μM, and 30 μM, with three replicates for each concentration. DMSO was used as the control wells, and cell-free medium was used as the blank wells. The cells were cultured at 37°C and 5% CO2 for another 48 h. Cell viability was detected using the CCK-8 assay kit. 10 μl of CCK-8 was added to each well, and the cells were incubated for 2 h. The absorbance at OD450 nm was then measured using a microplate reader.

[0034] Cell viability is calculated using the following formula:

[0035] Cell viability = [(As-Ab) / (Ac-Ab)] × 100%

[0036] As: Experimental wells (containing cell culture medium, CCK-8, and different concentrations of Bictegravir)

[0037] Ac: Compare with (culture medium containing cells, CCK-8, DMSO)

[0038] Ab: Blank wells (cell-free culture medium, CCK-8)

[0039] After calculating the cell viability at different concentrations of Bictegravir, bar charts were generated using GraphPad software.

[0040] See Figure 1 , Figure 1 The cell viability of THP-1 cells treated with different concentrations of Bictegravir was measured. The results showed that Bictegravir at concentrations of 0-30 μM did not significantly inhibit the activity of THP-1 cells.

[0041] Example 2:

[0042] Mycobacterium tuberculosis infection: THP-1 cells were used in 24-well plates at 5 × 10⁶ cells per well. 5 The number of cells was induced to become macrophages using the method described above. One hour before infection, 10 μM Bictegravir was added for pretreatment, and DMSO was added as a control. Mycobacterium tuberculosis H37Rv was used to infect the cells at an MOI of 10. After 4 hours, the cells were washed three times with PBS and cultured in 1640 complete medium at 37°C in a 5% CO2 incubator. During this process, Bictegravir or DMSO was continuously added.

[0043] CFU count by plating: 4 and 72 hours after infection, cells were lysed with 0.025% SDS and counted at 10⁻⁶ ppm. 2 10 3 Dilute the sample and plate it. Incubate at 37°C for approximately three weeks to count CFU.

[0044] See Figure 2 , Figure 2 The study investigated the inhibitory effect of 10 μM Bictegravir on Mycobacterium tuberculosis H37Rv at the THP-1 macrophage level. The results showed that Bictegravir could inhibit the growth of Mycobacterium tuberculosis in macrophages, and the survival rate of Mycobacterium tuberculosis was significantly reduced after the addition of Bictegravir compared with the DMSO control group.

[0045] Example 3:

[0046] Mycobacterium tuberculosis infection: THP-1 cells were used in 24-well plates at 5 × 10⁶ cells per well. 5 The number of cells was induced to become macrophages using the method described above. One hour before infection, 10 μM Bictegravir was added for pretreatment, and DMSO was added as a control. Mycobacterium tuberculosis H37Ra was used to infect the cells at an MOI of 10. After 4 hours, the cells were washed three times with PBS and cultured in 1640 complete medium at 37°C in a 5% CO2 incubator. During this process, Bictegravir or DMSO was continuously added.

[0047] CFU count by plating: 4 and 72 hours after infection, cells were lysed with 0.025% SDS and counted at 10⁻⁶ ppm. 2 10 3 Dilute the sample and plate it. Incubate at 37°C for approximately three weeks to count CFU.

[0048] See Figure 3 , Figure 3The study investigated the inhibitory effect of 10 μM Bictegravir on Mycobacterium tuberculosis H37Ra at the THP-1 macrophage level. The results showed that Bictegravir could inhibit the growth of Mycobacterium tuberculosis in macrophages, and the survival rate of Mycobacterium tuberculosis was significantly reduced after the addition of Bictegravir compared with the DMSO control group.

[0049] Example 4:

[0050] Minimum inhibitory concentration (MIC) assay: Take a 96-well plate. Add 100 μL ddH2O to each well on the outermost edge. Add 98 μL 7H9-OADC complete medium to each well of B2-G2. Add 50 μL 7H9-OADC complete medium to each of the remaining wells. Add 2 μL LB ictegravir to each well of B2-D2 and 2 μL LINH (isoniacinamide) to each well of E2-G2 as positive controls. Two-fold dilutions are applied to wells B2-G2 to B10-G10. Wells B11-G11 are negative controls without drug treatment. Finally, add 2 × 10⁻⁶ HCl to each well (except the ddH2O well). 5 A 50 μL solution of Mycobacterium tuberculosis H37Rv was sealed with sealing film and incubated at 37°C for 10-14 days. After incubation, the bacterial inhibition at different drug concentrations was observed, and the MIC values ​​were recorded.

[0051] Table 1 shows the minimum inhibitory concentrations (MICs) of Bictegravir and INH against Mycobacterium tuberculosis H37Rv. The results indicate that the minimum inhibitory concentration of Bictegravir against Mycobacterium tuberculosis in vitro is greater than 100 μM, significantly higher than the concentration used at the cellular level. This suggests that the anti-tuberculosis effect of Bictegravir at the macrophage level does not act directly on bacteria as an antibiotic, but rather inhibits intracellular survival of Mycobacterium tuberculosis by targeting the host and regulating the host's immune response. The minimum inhibitory concentration of INH against Mycobacterium tuberculosis is 0.02 μg / mL, consistent with the concentration range reported in the literature, serving as a positive control to demonstrate the reliability of the experimental data.

[0052] Table 1

[0053] Bictegravir >100μM Isoniazid (INH) 0.02 μg / mL

[0054] The above are merely preferred embodiments of the present invention and do not limit the patent scope of the present invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the patent protection scope of the present invention.

Claims

1. The use of Bictegravir as the sole active ingredient in the preparation of a medicament for inhibiting Mycobacterium tuberculosis H37Rv and / or H37Ra, characterized in that, The concentration of Bictegravir was 10 μM.

2. The use of Bictegravir as the sole active ingredient in the preparation of a medicament for treating tuberculosis caused by Mycobacterium tuberculosis H37Rv and / or H37Ra, characterized in that, The concentration of Bictegravir was 10 μM.

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