CN111658629A - Application of Fusariulin M and derivatives and pharmaceutically acceptable salts thereof - Google Patents

Abstract

The invention relates to the field of pharmacy, in particular to application of Fusariuin M and derivatives and pharmaceutically acceptable salts thereof, wherein the compound Fusariuin M has strong inhibitory activity on mycobacterium tuberculosis tyrosine phosphatase B and IC of the compound₅₀The concentration was 1.05. + -. 0.08. mu.M. The compound Fusariuelin M can effectively inhibit dephosphorylation activity of mycobacterium tuberculosis tyrosine phosphatase B, has low cytotoxicity, has obvious inhibition effect on proliferation of mycobacterium in macrophages, and has clinical application potential for tuberculosis treatment. In addition, the compound Fusarielin M is a natural metabolic activity small molecule product separated from microorganisms, the structure of the compound is analyzed, the compound can be produced by large-scale fermentation and separation, the source is rich, the production cost is low, and the possibility of drug formation is high.

Description

Application of Fusariulin M and derivatives and pharmaceutically acceptable salts thereof

Technical Field

The invention relates to the field of pharmacy, and in particular relates to Fusariuin M and application of derivatives and pharmaceutically acceptable salts thereof.

Technical Field

Tuberculosis is a chronic infectious disease caused by Mycobacterium tuberculosis (Mtb), one of the most prominent fatal infectious diseases worldwide. According to the world health organization 2019 report on global tuberculosis control, in 2018, there are 1000 million new patients, and about 120 million people die from Tuberculosis (TB). China is a country with high burden of tuberculosis and drug-resistant tuberculosis, and the research and development of anti-tuberculosis drugs are urgently!due to the continuous emergence of drug-resistant bacteria!

Mycobacterium tuberculosis tyrosine phosphatase B (MptpB) is a secreted virulence factor necessary for Mycobacterium tuberculosis to infect host cells, and has tyrosine phosphatase activity. MptpB is also a non-essential gene for the in vitro growth of Mtb, and deletion of MptpB does not affect the growth of Mtb in culture but severely inhibits the proliferation of Mtb in IFN-y activated macrophages and impairs the ability of Mtb to infect guinea pigs. MptpB has only 6% homology to human PTP1B, and lower homology increases the specificity of anti-tuberculosis drugs targeting MptpB. Therefore, the method for screening the antituberculosis drugs by taking the MptpB as a target point is an important strategy for developing a novel powerful low-toxicity antituberculosis drug.

Disclosure of Invention

The invention aims to overcome the problems in the prior art and provides application of Fusariuin M and derivatives and pharmaceutically acceptable salts thereof.

The purpose of the invention is realized by the following technical scheme:

the use of Fusarielin M and its derivatives or pharmaceutically acceptable salts thereof as tyrosine phosphatase inhibitors.

Use of Fusarielin M or a derivative or pharmaceutically acceptable salt thereof for the preparation of a medicament for inhibiting tyrosine phosphatase.

Preferably, the tyrosine phosphatase is mycobacterium tuberculosis tyrosine phosphatase B.

The use of Fusariuin M and derivatives or pharmaceutically acceptable salts thereof in the preparation of anti-tuberculosis drugs.

The molecular structural formula of the Fusarielin M is shown as the formula (I):

preferably, the concentration of the Fusarium M or the derivative or the pharmaceutically acceptable salt thereof is 0.95-1.12 mu M.

Compared with the prior art, the invention has the following technical effects:

the invention provides application of a compound Fusariuin M or a derivative or pharmaceutically acceptable salt thereof, wherein the compound Fusariuin M has strong inhibitory activity on mycobacterium tuberculosis tyrosine phosphatase B and IC of the compound₅₀The concentration was 1.05. + -. 0.08. mu.M. The compound Fusariuelin M can effectively inhibit dephosphorylation activity of mycobacterium tuberculosis tyrosine phosphatase B, has low cytotoxicity, has obvious inhibition effect on proliferation of mycobacterium in macrophages, and has clinical application potential for tuberculosis treatment. In addition, the compound Fusarielin M is a natural metabolic activity small molecule product separated from microorganisms, the structure of the compound is analyzed, the compound can be produced by large-scale fermentation and separation, the source is rich, the production cost is low, and the possibility of drug formation is high.

Drawings

FIG. 1 shows SDS-PAGE electrophoresis of purified Mycobacterium tuberculosis tyrosine phosphatase B protein; wherein lane M is a protein molecular weight standard (Thermo 26616), lanes 1 and 2 are purified MptpB (loading 1. mu.g of purified protein per well);

FIG. 2 IC inhibition of Mycobacterium tuberculosis tyrosine phosphatase B enzyme Activity by Fusariuin M₅₀A curve;

FIG. 3 identification of the type of inhibition of Mycobacterium tuberculosis tyrosine phosphatase B by the compound Fusariuin M;

FIG. 4 shows western blot assay results of the compound Fusariuilin M on intracellular MptpB-mediated activation of the MAPK pathway;

FIG. 5 intracellular Mycobacterium tuberculosis count at different concentrations of compound Fusariuin M treatment;

FIG. 6 shows the molecular structure of Fusariuin M.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be described in detail with reference to specific embodiments. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the examples given herein without any inventive step, are within the scope of the present invention.

Unless otherwise specified, the devices used in this example are all conventional experimental devices, the materials and reagents used are commercially available, and the experimental methods without specific descriptions are also conventional experimental methods.

Example 1

Expression and purification of Mycobacterium tuberculosis tyrosine phosphatase B (MptpB)

Escherichia coli BL21(DE3) T-strain expressing MptpB was inoculated into LB medium containing kanamycin (50. mu.g/mL) and shake-cultured at 37 ℃ overnight to obtain a culture. 2mL of the above culture was inoculated into 100mL of LB medium containing kanamycin and shake-cultured at 37 ℃ to OD₆₀₀Adding IPTG with final concentration of 0.1mM, shaking and culturing at 20 ℃, 180rpm overnight to induce the engineered bacteria to express protein MptpB, centrifuging the overnight-induced engineered bacteria expressing mycobacterium tuberculosis tyrosine phosphatase B (5000 × g, 4 ℃, 5min), collecting the bacteria, suspending the bacteria in 20mL of precooled lysis buffer (25mM Tris, 20mM imidazole, 500mM NaCl, pH 7.8), centrifuging again, repeating the above steps once to wash off the culture medium, performing the following steps on ice, precooling the used buffer and centrifuge to 4 ℃, resuspending the bacteria in 10mL of lysis buffer, adding TritonX-100 with final concentration of 1%, 5mM DTT, 1 × protease inhibitor, performing ultrasonic water bath for 15min to break the bacteria, performing centrifugation (11000rpm, 20 kDa, 4 ℃) on the broken bacteria, collecting the supernatant for purification, purifying by NiSepharose 6 Fast gel affinity chromatography, adding the supernatant to the nickel-based on the agarose gel chromatography, adding the nickel-loaded agarose column, washing the eluted protein with the eluent, washing, and eluting the protein eluting with 500 mL of 10 mM, and transferring the eluent to the eluent, wherein the eluent is 10 mM NaCl, and the eluent is 10 mMThe method comprises the steps of filtering a tube, centrifuging the tube for 15min at the temperature of 5000 × g and 4 ℃ for concentration, removing liquid in an outer-layer collecting tube, adding 5-10mL of preservation buffer (25mM Tris, 100mM NaCl, pH 7.8)5000 × g, centrifuging the tube for 15min at the temperature of 4 ℃ and repeating the steps for several times until imidazole in eluted protein is replaced, centrifuging the tube until the volume of the ultrafiltration concentrated liquid is 2mL, subpackaging the tube at the temperature of 4 ℃, determining the concentration of the purified MptpB protein according to a BCA protein quantification kit (Pierce), and analyzing the purified protein by SDS-PAGE electrophoresis to prove that the MptpB enzyme (shown in figure 1) with correct size and purity of more than 90 percent is obtained in the experiment.

Second, analysis of enzyme inhibitory Activity of Fusariulin M on Mycobacterium tuberculosis tyrosine phosphatase B (MptpB)

An inhibition experiment of the compound for inhibiting the mycobacterium tuberculosis tyrosine phosphatase B is carried out in a reaction buffer solution (50mM Tris, 100mM NaCl, pH7.0) by using the paranitrophenyl disodium phosphate (pNPP) as a substrate, and the mycobacterium tuberculosis tyrosine phosphatase B is the recombinant mycobacterium tuberculosis tyrosine phosphatase B protein obtained by the purification.

(1) Semi-inhibitory concentration IC₅₀Determination of value

The reaction was performed in a total volume of 200. mu.L in 3 parallel wells, and 1.5. mu.g of MptpB, a gradient of the compound (final concentration: 0. mu.M, 0.05. mu.M, 0.1. mu.M, 0.5. mu.M, 1. mu.M, 5. mu.M, 10. mu.M), the compound and MptpB were added to a 96-well plate in this order and premixed at room temperature for 5 min. After the incubation, pNPP was added to the final concentration of 1.3mM, the reaction was carried out at 37 ℃ for 5min, and the absorbance was read at 405nm for each well. At the same time, a blank (DMSO or compound only) was set up for background subtraction. The inhibitory activity of the compounds against the mppbb enzyme was calculated from the absorbance values according to the following calculation method: inhibition (%) [1- (experimental-blank)/(negative control-blank)]× 100% on the abscissa, the logarithm of the concentration of the compound and the inhibition rate are plotted on the ordinate, and IC is calculated by Origin Pro 8 software fitting₅₀Results are expressed as mean ± standard deviation.

(2) Inhibition type analysis

As described above, assay differentiationAbsorbance values corresponding to concentration gradients of pNPP substrate (0.1mM, 0.2mM, 0.4mM, 0.8mM, 1.6mM, 3.2mM) at compound concentrations (0. mu.M, 0.5. mu.M, 1. mu.M, 2.5. mu.M), and double reciprocal plots were performed with the reciprocal of the substrate concentration as the abscissa and the reciprocal of the reaction rate as the ordinate, to determine the type of inhibition and to calculate the inhibition constant K_i. Each sample was assayed in triplicate.

(3) Enzyme inhibition specificity assay for compounds

The enzyme inhibition activity of the compound on homologous proteins of mycobacterium tuberculosis tyrosine phosphatase A (MptpA) and human tyrosine phosphatase 1B (PTP1B) is determined, and the reaction system is the same as MptpB. Differently, 0.75. mu.g of MptpA was added to the reaction, the compound concentrations were 0. mu.M, 1.5625. mu.M, 3.125. mu.M, 6.25. mu.M, 12.5. mu.M, 25. mu.M, 50. mu.M, and the compound was preincubated with MptpA for 5min before adding the substrate at a final concentration of 1.2 mM; mu.g of PTP1B was added to the reaction at compound concentrations of 0. mu.M, 1.5625. mu.M, 3.125. mu.M, 6.25. mu.M, 12.5. mu.M, 25. mu.M, 50. mu.M, and 2mM substrate was added to the final concentration after 5min of preincubation of the compound with PTP 1B. Each sample was assayed in 3 parallel wells and the half inhibitory concentration IC of compound against MptpA and PTP1B was calculated₅₀。

(4) Results of the experiment

The half-inhibitory concentration IC of the compound Fusariuilin M on mycobacterium tuberculosis tyrosine phosphatase B is determined by experiment₅₀1.05. + -. 0.08. mu.M (see Table 1, FIG. 2); the compound has better specificity to MptpB enzyme inhibition, and has selectivity of more than 15 times compared with homologous proteins of mycobacterium tuberculosis MptpA and human PTP1B (see Table 1). Further characterization of the type of inhibition of MptpB by compound Fusariuin M with four straight lines intersecting the vertical axis (see FIG. 3) at different concentrations of inhibitor indicated that compound Fusariuin M is a competitive inhibitor, and simultaneous determination of the inhibition constant K for compound Fusariuin M_iIt was 1.03. + -. 0.39. mu.M (Table 2).

TABLE 1 results for in vitro enzyme inhibitory Activity of the Compound Fusariuin M (IC)₅₀，μM)^a

SI＝selectivity index.SI^*＝IC₅₀ ^MptpA/IC₅₀ ^MptpB；SI^**＝IC₅₀ ^PTP1B/IC₅₀ ^MptpB.

^aThe experiment was repeated three times and the results are expressed as mean ± standard deviation.

TABLE 2 measurement of MptpB enzyme inhibition constant by Fusarelin M compound (K)_i，μM)^a

Compound (I)	MptpB(Ki，μM)
		Fusarielin M	1.03±0.39

^aThe experiment was repeated three times and the results are expressed as mean. + -. standard deviation

Cell proliferation toxicity test of compound Fusariuelin M

(1) Cell culture

The human lung cancer cell strain A549 is cultured in RPMI 1640 culture medium containing 10% FBS and double antibody (penicillin and streptomycin), and mouse alveolar macrophage J774A.1 and human hepatoma cell HepG2 are cultured in DMEM culture medium containing 10% FBS and double antibody (penicillin and streptomycin).

(2) Cell plating

When the adherence concentration of mouse alveolar macrophage J774A.1 reaches 80% -90%, abandoning the culture medium, washing the cells twice with PBS, adding pancreatin for digestion, terminating digestion of the culture medium, transferring to a centrifuge tube, centrifuging at 1000rpm for 3min, abandoning the supernatant, suspending the cells with the culture medium, taking a small amount of cell suspension for counting, adjusting the density of the cell suspension to 5 × 10 with the culture medium⁴cells/mL, 0.1 mL/well diluted cells were inoculated96-well plates. Place the cell culture plate in CO at 37 ℃₂And (5) an incubator for overnight culture.

(3) Treatment of compounds

Culture medium (25, 50, 100, 120, 150, 180, 200 μ M) containing different concentrations of compounds was added to the 96-well plates inoculated with cells, while a negative control (1% DMSO in combination) and a blank control (medium only) were set up, three parallel wells. Place the cell culture plate in CO at 37 ℃₂And (5) culturing for 72 hours in an incubator.

(4) MTS processing

The medium was discarded from the wells, 0.1mL of medium and 20. mu.L of MTS (promega) were added to each well, and the culture was continued for 0.5-1 h. The plates were removed and the absorbance of each well was measured at 490 nM. The inhibition activity of the compound treatment with different concentrations on cell proliferation is calculated according to the absorbance value according to the following calculation method: inhibition (%) [1- (experimental-blank)/(negative control-blank)]× 100% in each case, the logarithm of the concentration of the compound is plotted on the abscissa and the inhibition ratio on the ordinate, and iC is calculated by Origin Pro 8 software fitting₅₀Results are expressed as mean ± standard deviation.

(5) Results of the experiment

The experimental result shows that the compound Fusariuelin M has no strong cytotoxicity to human lung cancer cell strain A549, human liver cancer cell strain HepG2 and mouse macrophage strain J774A.1, and iC₅₀The values were all greater than 100. mu.M (see Table 3).

TABLE 3 cell proliferation toxicity test results (IC) for compound Fusariuin M₅₀，μM)

Cell line	IC of the Compound50Value (μ M)
		Human lung cancer cell A549	120.16±3.12
Human liver cancer cell HepG2	169.84±6.29
		Mouse macrophage J774A.1	127.03±2.06

Example 2

Determination of in vitro anti-tubercle bacillus growth activity of compound Fusariuelin M

The experiment adopts resazurin as an indicator reagent for bacterial growth, the in vitro antibacterial activity of a compound Fusarielin M on 4 strains of mycobacterium (m.smegmatis, m.marinum, m.bovis, m.tubericosis), 1 strain of Staphylococcus aureus (Staphylococcus aureus) and 1 strain of Escherichia coli (Escherichia coli) is measured, and the antibacterial activity of the compound is evaluated by taking Minimum Inhibitory Concentration (MIC) as an index.

(1) Bacterial strains

The mycobacterium: tuberculosis H₃₇Ra，M.bovis BCG，M.smegmatis mc²155, m. marinumbaa-535, in 7H9 (containing 0.2% glycerol, 0.05% Tween-80, 10% OADC) liquid medium (BD).

Staphylococcus aureus (Staphylococcus aureus ATCC6538) and Escherichia coli (Escherichia coli ATCC25922) were cultured in LB medium.

(2) The in vitro bacteriostatic activity of the compound was tested using resazurin (aladdin) as an indicator of bacterial growth. In a 96-well plate, a gradient dilution of the compound (100. mu.L) was co-cultured with the bacteria (100. mu.L) for a certain period of time, then 30. mu.L of a 0.01% resazurin solution was added to each well, the incubation was continued overnight, the fluorescence value of each well (excitation wavelength 530nm, emission wavelength 590nm) was measured, and the Minimal Inhibitory Concentration (MIC) of the compound against each bacteria was calculated.

(3) The experimental results are as follows: compound Fusariuilin M for binding mycobacterium tuberculosis H₃₇The Ra strain has good in-vitro antibacterial activity, the MIC value is 12.3mg/L, and the resistance to golden yellow grape ballsThe bacteria had a weak bacteriostatic activity (MIC of 19mg/L), but had no significant in vitro growth inhibitory activity against mycobacterium smegmatis, mycobacterium marinum, mycobacterium bovis and escherichia coli (see table 4).

TABLE 4 in vitro antibacterial Activity (MIC, mg/L) of the Compound Fusariuin M

Second, analysis of intracellular MptpB inhibitory Activity of Compound Fusariulin M

When tubercle bacillus infects cells, the virulence protein MptpB can inhibit apoptosis and down regulate immune response by activating cell Akt signals, inhibiting activation of ERK1/2 and p 38. If compound Fusariuelin M is able to inhibit intracellular MptpB, it is possible to reverse the regulation of the cellular pathway by MptpB, leading to the opposite change in the above-mentioned criteria, i.e.a decrease in the activation of the Akt pathway and an increase in the activation of ERK1/2 and p 38. Accordingly, MPtpB (wild type) and MPtpB/C160S (enzyme activity deletion type) were transiently transfected into macrophage Raw264.7, the cell immunity was stimulated with interferon IFN-. gamma.and compounds were added for treatment, and total cell protein was collected and extracted, and Western blot was performed to examine the protein expression amounts of pERK1/2, ERK1/2, pp38, and p38 in the cells.

(1) Mouse macrophage Raw264.7 culture

Mouse macrophage Raw264.7 was cultured in DMEM medium containing 10% FBS and diabodies (penicillin and streptomycin) at 37 deg.C in CO₂Culturing in incubator, when the convergence of cell adherent growth reaches 80%, performing passage and plating, and performing 3 × 10 a day before cell transfection⁶Cells were plated into 6-well cell culture plates per well. Place the cell culture plate in CO at 37 ℃₂The incubator was overnight to allow the cells to adhere to the wall sufficiently.

(2) Transient transfection of cells to express MptpB

150. mu.L of Opti-MEM containing 3ug of expression plasmid (MptpB, MptpB/C160S, Vector) was prepared, and 9. mu. LFu was added

HD TransThe transfection Reagent mixture was mixed gently and incubated at room temperature for 15min to form a plasmid-transfection Reagent mixture. Slowly dripping the transfection compound into the cells, slightly shaking and uniformly mixing, and culturing in an incubator for 24h to express the target protein.

(3) Treatment of compounds

After 24h of transfection, fresh medium (0,10, 20. mu.M) containing different concentrations of the compound was replaced and incubated at 37 ℃ for 1 h. IFN-gamma with a final concentration of 40ng/mL is added to each well, and cells are collected for protein extraction after further 2h of culture.

(4) Extraction of Total cellular protein

The culture supernatant was discarded and the cells were washed twice with PBS (2 mL/well). Add IP lysine/washbuffer (containing 1 Xprotease inhibitor, Pierce) at 200. mu.L/well, mix well, and place on ice to lyse for 15 min. 13000 Xg, centrifuging for 10min at 4 ℃, taking supernatant to carry out protein concentration determination and Western blot experiment.

Detecting an antibody: rabbit anti-p-ERK 1/2(Thr202/Tyr204), rabbit anti-ERK 1/2, rabbit anti-p 38(Thr180/Tyr182), rabbit anti-p 38 purchased from CST corporation; murine anti-alpha-tubulin, a goat anti-mouse IgG secondary antibody labeled with Horse Radish Peroxidase (HRP), and a goat anti-rabbit secondary antibody labeled with HRP were purchased from Proteintech.

(5) The experimental results are as follows:

MptpB can inhibit the activation of MAPK pathway in cells under the stimulation of IFN-gamma, and reduce the phosphorylation level of p38 and ERK 1/2; MptpB was dependent on the phosphatase activity for inhibition of the MAPK pathway, so the point mutation deletion enzyme MptpB/C160S lost inhibition of MAPK, consistent with transfection unloading (FIG. 4). Therefore, in this experiment, the inhibitory effect of MptpB on intracellular MAPK pathway was consistent with literature reports. After the compound Fusariulin M inhibits the intracellular MptpB, p-p38 and p-ERK1/2 are increased and are concentration-dependent; in cells transfected with MptpB/C160S deleted by the empty and point mutant enzymes, the compound Fusariulin M did not affect the expression levels of-pp 38 and p-ERK1/2 (FIG. 4). Thus, Fusarielin M was able to block the inhibitory effect of MptpB on the intracellular MAPK pathway.

Growth inhibitory Activity of Compound Fusariuin M on Mycobacterium macrophage

In view of the good in vitro and in vivo MptpB inhibitory activity of the compound Fusariuin M, the anti-tubercular activity of the compound at the cellular level was further investigated. Mouse alveolar macrophage J774A.1 was used as a host cell, and the cell was infected with Mycobacterium bovis (M.bovis BCG) strain, and the activity of Fusariuin M in inhibiting the proliferation of M.intracellulare was measured.

(1) Culture of Mycobacterium bovis M.bovis BCG strain

7H9 (containing 0.2% glycerol, 0.05% Tween-80, 10% OADC) liquid medium, 7H10 (containing 0.2% glycerol, 10% OADC) solid medium (BD).

Inoculating frozen Mycobacterium bovis BCG strain at-80 deg.C to 7H10 solid culture medium, and performing inverted culture at 37 deg.C for 2-3 weeks until a layer of lawn grows on the plate, and can be used for cell infection or passage.

(2) J774A.1 cell culture and plating

Mouse alveolar macrophage J774A.1 is cultured in DMEM medium containing 10% FBS and double antibody (penicillin and streptomycin) and placed in CO at 37 DEG C₂Culturing in incubator, when the convergence of cell adherent growth reaches 80%, performing passage and plating, one day before cell infection, according to 2 × 10⁵Per well, cells were plated into 24-well cell culture plates. Place the cell culture plate in CO at 37 ℃₂The incubator was overnight to allow the cells to adhere to the wall sufficiently.

(3) Infection of cells

Scraping 2-3 weeks old BCG lawn into 50mL centrifuge tube, resuspending with 20mL preheated PBS buffer solution, adding 5mL glass beads, vortex shaking for 10min, centrifuging at 600rpm for 10min, sucking supernatant, and diluting the bacterial solution to 1 × 10% with 10% bovine serum-containing DMEM culture medium⁶CFU/mL, 200 μ L/well of diluted bacterial suspension was added to a 24-well cell culture plate (MOI ═ 1), and cultured for 4 hours. After 4h of infection, the culture medium was discarded, and the cells were washed 2 times with 1 mL/well of pre-warmed PBS, to wash out bacteria that were not engulfed into the cells. 2mL of cell culture medium containing different concentrations of compound or 0.8% DMSO were added, respectively, and incubation was continued for 72 h. Each experimental group was provided with 3 parallel wells.

After three days, the cell culture supernatant was carefully discarded, and the cells were washed 2 times with pre-warmed PBS at 1 mL/well. 0.2mL of 0.05% SDS was added to each well, and the mixture was allowed to stand at 37 ℃ for 5min to lyse the cells and release viable intracellular bacteria. Transferring the cell lysate to a 1.5mL centrifuge tube, vortexing and shaking for 5min, fully lysing cells and resuspending thalli. And (3) diluting the lysate by 100 times by using 7H9 culture medium, coating 200 mu L of the diluent on a 7H10 plate, performing inverted culture at 37 ℃ for 3-4 weeks, counting growing colonies on the plate, and calculating the bacteriostasis rate.

(4) Results of the experiment

The compound Fusariuelin M has an inhibiting effect on the proliferation of BCG in macrophage J774A.1, and is in a compound concentration dependence, as shown in figure 5, when the compound concentrations are respectively 5 muM, 10 muM and 20 muM, the number of viable bacteria in cells is respectively reduced by 42%, 58% and 62%. Previous experiments prove that 40 mu M of compound Fusariuelin M has no obvious cytotoxicity on J774A.1, so the intracellular bacteriostatic effect observed here is caused by the antibacterial activity of the compound, but the apoptosis is caused by the toxicity of the compound, and the viable count is reduced due to the damage to the living environment of bacteria.

Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention, and not for limiting the protection scope of the present invention, although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims (5)

1. Use of Fusarielin M or a derivative or pharmaceutically acceptable salt thereof as a tyrosine phosphatase inhibitor.
2. Use of Fusarielin M or a derivative or pharmaceutically acceptable salt thereof for the preparation of a medicament for inhibiting tyrosine phosphatase.
3. 3. The use according to claim 1 or 2, said tyrosine phosphatase being mycobacterium tuberculosis tyrosine phosphatase B.
4. Use of FusariuelinM and derivatives or pharmaceutically acceptable salts thereof in the preparation of anti-tuberculosis drugs.
5. 5. The use of any one of claims 1, 2 or 4, wherein the concentration of Fusarium M or its derivative or pharmaceutically acceptable salt thereof is 0.95-1.12 μ M.

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