CN111973598B

CN111973598B - Application of dabrafenib and/or pharmaceutically acceptable derivatives thereof in preparation of anti-saxavirus drugs

Info

Publication number: CN111973598B
Application number: CN202010939268.7A
Authority: CN
Inventors: 徐国东; 刘愈杰; 幸晓莹
Original assignee: Canvest Wuhan Biotechnology Co ltd
Current assignee: Canvest Wuhan Biotechnology Co ltd
Priority date: 2020-09-09
Filing date: 2020-09-09
Publication date: 2021-04-13
Anticipated expiration: 2040-09-09
Also published as: CN111973598A

Abstract

The invention provides application of dabrafenib and/or pharmaceutically acceptable derivatives thereof in preparation of a medicament for resisting a sandparticle virus. The invention detects the cytotoxicity of dabrafenib on Vero cells and BHK-21 cells and the antiviral activity of dabrafenib on arenaviruses, shows that the dabrafenib has stronger effect of resisting the arenaviruses at a cellular level, and provides a new direction for preventing and treating arenavirus infectious diseases.

Description

Application of dabrafenib and/or pharmaceutically acceptable derivatives thereof in preparation of anti-saxavirus drugs

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to application of dabrafenib and/or pharmaceutically acceptable derivatives thereof in preparation of a medicine for resisting arenavirus.

Background

Mammalian arenaviruses (Mammarenavirus) belong to the Arenaviridae (Bunyavirales) family of bunyaviridae (Arenaviridae), and consist mainly of old and new world arenaviruses, including many virulent viruses that pose a significant threat to humans, such as Lassa virus (LASV), vernonia virus (Jun virus, JUNV), machupulo virus (Machupo virus, MACV), and the like. Wherein LASV is prevalent in West Africa countries such as Nigeria, Libiria, Serioleon, and Guinea, where 10-30 million people infect each year and about 5000 people die. JUNV is prevalent mainly in the argentina Pampa area, causing argentina hemorrhagic fever with a mortality rate of 15% -30%. In addition, Lymphocytic choriomeningitis virus (LCMV) exists in our country, mainly in mice, and humans are infected by contact with mouse excreta and outbreaks in immunocompromised individuals, causing central nervous system infections, one of the important pathogens that has been overlooked. Among the pathogenic microorganisms of the interpersonal infection established by the ministry of health of the people's republic of China, the arenavirus family accounts for 8.

Arenaviruses are widespread and seriously threaten human health, and there is currently no FDA-approved vaccine or drug specific for arenaviruses. The current main treatment method is mainly limited to ribavirin, but the treatment effect is limited and the ribavirin has strong side effects. Therefore, it is of great importance to research and develop effective anti-arenavirus infection drugs.

Dabrafenib is a drug for the treatment of skin cancer melanoma. The Food and Drug Administration (FDA) approved trametinib in combination with dabrafenib for the treatment of patients with unresectable (inoperable) and metastatic advanced melanoma on 1/10 d/2014. In 8 months of 2018, the joint research group of university of shenhu, osaka university, and tokyo university in japan showed that a pharmaceutical ingredient capable of inhibiting neuronal necrosis was found in "dalafenib", or that it will be a therapeutic agent for parkinson's disease.

Disclosure of Invention

The invention aims to solve the problems of limited treatment effect and strong side effect of the existing anti-arenavirus medicine.

Therefore, the invention provides the application of the dabrafenib and/or the medicinal derivatives thereof in preparing the anti-arenavirus medicament.

Further, the arenavirus is lymphocytic choriomeningitis virus.

Further, the anti-sandparticle virus activity concentration of the dabrafenib is 0.5-128 mu M.

Further, the anti-saxavirus medicine comprises an active ingredient and pharmaceutically acceptable auxiliary materials, wherein the active ingredient comprises dabrafenib and/or pharmaceutically acceptable derivatives thereof.

In the invention, the dalafenib (trade name: Tafinalar, common name: Dabrafinib) is an existing drug for treating skin cancer melanoma, the active ingredient of the dalafenib is dalafenib mesylate, and the chemical structural formula is as follows:

。

the invention has the beneficial effects that:

the invention detects the cytotoxicity of dabrafenib on Vero cells and BHK-21 cells and the antiviral activity of dabrafenib on arenaviruses, shows that the dabrafenib has stronger effect of resisting the arenaviruses at a cellular level, and provides a new direction for preventing and treating arenavirus infectious diseases.

The present invention will be described in further detail below with reference to the accompanying drawings.

Drawings

FIG. 1 is a graph of cell viability on Vero cells for different concentrations of dabrafenib in example 1 of the present invention;

FIG. 2 is a graph showing the cell viability of dabrafenib on BHK-21 cells at various concentrations in example 1 of the present invention;

FIG. 3 is a graph of cell viability on Vero cells for different concentrations of ribavirin according to example 1 of the present invention;

FIG. 4 is a graph of cell viability for different concentrations of ribavirin on BHK-21 cells in example 1 of the present invention;

FIG. 5 is a graph of inhibition of LCMV viral replication on Vero cells at various concentrations of dabrafenib in example 2 of the invention;

FIG. 6 is a graph of the inhibition rate of different concentrations of dabrafenib on BHK-21 cells for inhibiting LCMV viral replication in example 2 of the present invention;

FIG. 7 is a graph of inhibition of LCMV viral replication on Vero cells by different concentrations of ribavirin in example 2 of the invention;

FIG. 8 is a graph of the inhibition rate of different concentrations of ribavirin on BHK-21 cells for LCMV viral replication in example 2 of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1:

this example investigated the cytotoxicity of the target compound, dabrafenib, in the following specific procedure:

BHK-21 cells and Vero cells were 1.5X 10 cells, respectively⁴Cells/well were seeded in 96-well cell culture plates at 37 ℃ in 5% CO₂After culturing for 12-16 h in an incubator, firstly changing a DMEM culture medium containing 2% FBS, then treating with Dalafinil with the concentration of 0.5 MuM, 1 MuM, 2 MuM, 4 MuM, 8 MuM, 16 MuM, 32 MuM, 64 MuM and 128 MuM, adding DMSO with the same volume into a negative control, setting three multiple wells for each concentration, incubating for 36h at 37 ℃, and simultaneously setting a zero adjusting well only with the culture medium without cells. Discarding the supernatant, washing with PBS once, adding 50 μ l of 0.5% MTT PBS solution into each well, standing at 37 deg.C for 4h in a dark condition, observing blue-purple crystals on the bottom of the plate, terminating the culture, carefully sucking the supernatant, adding 100 μ l DMSO into each well, and placing on a shaking table to slowly shake for 10min to fully dissolve the crystals. The absorbance of each well was measured using a microplate reader at OD570nm, the cytotoxicity of each well was calculated from the values of DMSO control wells after zeroing the wells, as shown in FIGS. 1 and 2, respectively, and the CC of Dalafinil on Vero and BHK-21 cells was calculated by plotting the cell viability curves at different concentrations₅₀88.87 μ M and 106.3 μ M, respectively.

In addition, the comparative experiments of treating BHK-21 cells and Vero cells with ribavirin were conducted in the same manner as described above with respect to the dabrafenib treatment except that the concentrations of ribavirin were 1. mu.M, 3.3. mu.M, 10. mu.M, 33. mu.M, 100. mu.M, 330. mu.M and 1000. mu.M; the absorbance of each well was measured using a microplate reader at OD570nm, the cytotoxicity of each well was calculated from the values of the DMSO control wells after zeroing the wells, as shown in FIGS. 3 and 4, respectively, and the CC of ribavirin on Vero cells and BHK-21 cells was calculated by plotting the cell viability curves at different concentrations₅₀Are respectively as>1000. mu.M and 100.4. mu.M.

Example 2:

this example studies the antiviral activity of the target compound dabrafenib on arenavirus (LCMV) by the following specific procedure:

BHK-21 cells and Vero cells were 1.5X 10 cells, respectively⁴Cells/well were seeded in 96-well cell culture plates at 37 ℃ in 5% CO₂After culturing for 12-16 h in the incubator, the culture medium is replaced by DMEM containing 2% FBS, the Dalanib with the concentration of 0.5 mu M, 1 mu M, 2 mu M, 4 mu M, 8 mu M, 16 mu M, 32 mu M, 64 mu M and 128 mu M is used for treatment, the same volume of DMSO is added into a negative control, and three multiple wells are arranged at each concentration. After 1h incubation, cells were infected with LCMV. After 36h, the effect of dabrafenib on inhibiting virus replication was tested by measuring the titer of the supernatant, and the results on two cells are shown in fig. 5 and fig. 6, respectively, and the compound dabrafenib has a significant inhibition effect on LCMV under different concentration conditions. By drawing the inhibition rate curve of the rafenib inhibiting virus under different concentrations, the IC of the darafinil inhibiting LCMV in Vero and BHK-21 cells is calculated₅₀3.863 μ M and 15.99 μ M, respectively.

In addition, the comparative experiment was conducted with respect to antiviral activity of ribavirin against arenavirus (LCMV), and the treatment process was substantially identical to that of the above-described Darafenib treatment process except that the treatment concentrations of ribavirin were 1. mu.M, 3.3. mu.M, 10. mu.M, 33. mu.M, 100. mu.M, 330. mu.M and 1000. mu.M. The results of the inhibition of virus replication by ribavirin are shown in FIGS. 7 and 8, respectively, and the IC of ribavirin on Vero cells and BHK-21 cells was calculated by plotting the inhibition rate curves of ribavirin against virus at different concentrations₅₀40.13 μ M and 4.26 μ M, respectively.

The above examples are merely illustrative of the present invention and should not be construed as limiting the scope of the invention, which is intended to be covered by the claims and any design similar or equivalent to the scope of the invention.

Claims

1. Application of dabrafenib in preparing medicines for resisting lymphocyte choriomeningitis virus.

2. The use of claim 1, wherein: the active concentration of the dalafenib anti-lymphocyte choriomeningitis virus is 0.5-128 mu M.

3. The use of claim 1, wherein: the lymphocyte choriomeningitis virus resisting medicine comprises an active ingredient and pharmaceutically acceptable auxiliary materials, wherein the active ingredient comprises dabrafenib.