KR102602837B1 - An antipathogenic compound - Google Patents

Abstract

The present invention relates to a novel pyrazole-based compound that has selective antibacterial activity against Gram-negative pathogens having OxyR, an active oxygen sensor. In particular, the present invention relates to a novel anti-virulence compound targeting OxyR, an active oxygen sensor required for virulence of Pseudomonas aeruginosa, an opportunistic human pathogen.

Description

An antipathogenic compound}

The present invention relates to a novel pyrazole-based compound that has selective antibacterial activity against Gram-negative pathogens that have OxyR as an active oxygen sensor. In particular, the present invention relates to a novel compound that has anti-virulence activity by targeting OxyR, which is essential for the pathogenicity of Pseudomonas aeruginosa, a human opportunistic pathogen.

Antibiotics have brought about a major revolution in the treatment of bacterial infectious diseases. However, some pathogenic bacteria have begun to develop resistance to antibiotics over time. In particular, there are cases where multidrug resistance occurs, in which bacteria have more systematic and widespread resistance to two or more antibiotics.

The rapid evolution and spread of antimicrobial resistance (AMR) poses a significant threat to public health around the world. Considering the complex nature of infectious diseases and the fluid pathophysiology of the host environment, it is becoming increasingly difficult to treat infectious diseases using traditional antibacterial agents. To address AMR and its related complex infectious diseases, multidimensional efforts are needed to discover and develop new antibacterial agents in a way that mitigates the emergence of resistance to antimicrobial agents. As part of this effort, it is important to note that although it does not affect bacterial growth, it does not affect bacterial pathogenicity. Research to develop antivirulents (antipathogenics, antivirulents) is actively underway.

Structural details of the OxyR peroxidesensing mechanism, PNAS (Proceedings of the National Academy of Sciences of the United States of America) 2015, 19;112(20):6443-6448.

The purpose of the present invention is to develop a candidate selective antibacterial or antiviral agent against Gram-negative pathogens that has OxyR as an active oxygen sensor.

The present invention provides a pyrazole-based compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof.

[Formula I]

The present invention provides an antibacterial or anti-venom agent against bacteria containing a pyrazole-based compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof.

[Formula I]

In the present invention, the compound of formula (I) has excellent anti-virulence activity under infection conditions against Gram-negative pathogens that have OxyR as an active oxygen sensor. The compound of formula I reduces the viability of Gram-negative pathogens by targeting OxyR, an active oxygen sensor required for the virulence of Gram-negative pathogens.

Figure 1 is a diagram showing that the OxyR function of Pseudomonas aeruginosa (PA14, WT) is inhibited by a compound of formula I (comp I), thereby inhibiting continuous dilution growth of Pseudomonas aeruginosa under aerobic conditions.
Figures 2a and 2b are diagrams showing a docking model for how a compound of Formula I binds to the OxyR regulatory site.
Figure 3 is a diagram showing how (a) the oxidation-reduction state of OxyR and (b) the transcription of katA , an OxyR target gene, are changed by a compound of Formula I.
Figure 4 is a diagram showing the change in mortality rate of fruit flies infected with Pseudomonas aeruginosa PA14 or Staphylococcus aureus SA3 by compounds of formula I.

In the present invention, the compound of formula (I) has excellent anti-virulence activity against Gram-negative pathogens that have OxyR as an active oxygen sensor. In particular, it can exhibit anti-virulence activity by interfering with the process necessary for bacteria to cause disease in an infected host.

This anti-virulence strategy of the present invention is a new approach in that it inhibits the virulence factors that bacteria use to cause disease in an infected host. Unlike existing antibiotics, these can specifically inhibit the virulence of infected pathogens rather than directly targeting bacterial growth. For this reason, the advantage of the anti-virulence compound of the present invention is that it is unlikely to promote the evolution of bacterial resistance, which inevitably occurs in the process of inhibiting bacterial growth.

Additionally, unlike existing antibiotics that damage the body's normal microbial flora, it selectively inhibits the virulence of specific Gram-negative pathogens, so it has the advantage of reducing such concerns. Until recently, there have been results on anti-virulence agents targeting bacterial virulence factors such as quorum sensing and secretion systems, but the compound of formula I is the first anti-virulence agent discovered by targeting OxyR, an important virulence factor. It is an OxyR compound.

When infected with pathogens, the human body attacks the pathogens in various ways, and the pathogens respond in various ways. Since free radicals are the main attack substances produced by the body's immune cells, all pathogens have a defense system that can survive attacks by free radicals.

That is, in Gram-negative pathogens that have OxyR as an active oxygen sensor, OxyR is a regulator that recognizes H ₂ O ₂ , a type of active oxygen generated from oxygen, and regulates the expression of antioxidant proteins, and is a regulator that regulates the expression of antioxidant proteins in bacteria under conditions where active oxygen is present. It also allows you to survive. OxyR is recognized as an important virulence factor because it is essential for the opportunistic bacterium Pseudomonas aeruginosa to survive within the host after infection. In particular, OxyR regulates the transcription of KatA, a catalase important for acute virulence and intrinsic AMR in some Pseudomonas aeruginosa strains.

The regulatory mechanism of OxyR can be explained by structural changes in the redox switch based on conserved cysteine residues involved in ROS binding. The present invention sought to develop a new anti-virulence candidate targeting OxyR by identifying a new compound that inhibits the function of OxyR in response to active oxygen.

The present invention performed computer-based virtual screening to identify new compounds that bind to the OxyR regulatory region based on the known structural and functional characteristics of OxyR of Pseudomonas aeruginosa, a Gram-negative pathogen. As a result, the compound of Formula I of the present invention was derived. did. By measuring the aerobic dilution sensitivity, which is the phenotype of the oxyR deletion mutant, among the effective substances that cause loss of function of OxyR, the compounds of the present invention affect both the transcriptional regulation induced by OxyR and the redox state related to disulfide bond formation. It was confirmed to have an effect. Most importantly, the compounds of the present invention selectively attenuated the pathogenicity of Pseudomonas aeruginosa, especially in the Drosophila systemic infection model.

Therefore, in the present invention, the compound of formula I can be used as an antibacterial or anti-venom agent against Gram-negative pathogens that have OxyR as an active oxygen sensor. The compound of Formula I can be produced through organic synthesis using commercially available raw materials.

In one embodiment, the pharmaceutically acceptable salt of the compound represented by Formula I of the present invention may be a salt of a carboxyl group, a metal ion, ammonia, and an organic base.

Specifically, metal salts include alkali metal salts, including sodium, potassium, and lithium salts, or alkaline earth metal salts, including calcium, magnesium, and barium salts. Salts with organic bases include salts with amines, especially aliphatic amines such as trimethylamine, triethylamine, cyclohexylamine, tert-butylamine, N-(phenylmethyl)benzene-ethylamine, N,N'-di. Benzyl ethylenediamine, choline, 2-(dimethylamino)ethanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine , morpholine, 4-(2-hydroxy-ethyl)morpholine, piperazine, 1-(2-hydroxyethyl)pyrrolidine, triethanolamine, 2-amino-2-(hydroxymethyl)propano -1,3-diols, amino acids such as arginine, lysine, histidine or ornithine, aromatic amines such as aniline, methylaniline, naphthyl-amine, or heterocyclic amines such as for example 1H-imidazole or pyridine. Includes.

The present invention provides an anti-venom agent against Gram-negative pathogens, which includes a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof, and has OxyR as an active oxygen sensor.

[Formula I]

Below, preferred embodiments are presented to aid understanding of the present invention. However, the following examples are provided only to make the present invention easier to understand, and the content of the present invention is not limited by the examples.

Example 1. Synthesis of 1-benzyl-4-(2-mecaptoacetamido)-1H-pyrazole-3-carboxylic acid (Formula I)

A compound of Formula I was synthesized through the same process as Scheme 1 below.

Scheme 1.

1-1. Preparation of 2-(tritylthio)acetic acid (Compound 2)

Trifluoroacetic acid was added to a solution of mercaptoacetic acid (2.30 g, 1.74 mL, 25 mmol, 1 eq) and triphenylmethanol (6.5 g, 25 mmol, 1 eq) in methylene chloride (50mL). (5 ml) was added over 5 minutes. After stirring at room temperature for 1 hour, the solvent was removed under vacuum and the product was obtained. The obtained product was recrystallized using a 1/2 solvent of CH ₂ Cl ₂ /Hexane to obtain 2-tritylthioacetic acid (7.52 g, yield 90%) as a white solid.

1-2. Synthesis of methyl 4-nitro-1H-pyrazole-3-carboxylate (Compound 4)

A solution of 4-nitro-1H-pyrazole-3-carboxylic acid (Compound 3, 4-nitro-1H-pyrazole-3-carboxylic acid, 2.5 g, 15.92 mmol, 1 eq) in methanol (30 mL) was dissolved in H ₂ SO ₄ ( 1.7 mL, 3.12 g, 31.84 mmol, 2 eq) was added and refluxed for 1 hour. Afterwards, the reaction solution was concentrated, dissolved in EtOAc, and washed with saturated NaHCO ₃ and brine. The organic layer was mixed and dehydrated with Na ₂ SO ₄ and the solvent was removed under vacuum to obtain methyl 4-nitro-1H-pyrazole-3-carboxylate (2.66 g, 97.79%) as a white solid.

1-3. Synthesis of methyl 1-benzyl-4-nitro-1H-pyrazole-3-carboxylate (compound 5)

An acetone solution of methyl 4-nitro-1H-pyrazole-3-carboxylate (Compound 4, 2.62 g, 15.32 mmol, 1 eq) was added with potassium carbonate (3.70 g, 26.81 mmol, 1.75 eq) at a temperature of 0 °C. Benzyl bromide (3.14 g, 18.38 mmol, 1.2 eq) was added. The reaction solution was stirred at room temperature for 12 hours. Afterwards, the reaction solution was filtered using Celite, and the filtrate was concentrated under reduced pressure. Impurities were removed from the residue using gel chromatography (using a 20% hexane solvent in EtOAc), and methyl 1-benzyl-4-nitro-1H-pyrazole-3-carboxylate (3 g, 53.19%) was obtained as a pale yellow solid. ) was obtained.

1-4. Synthesis of methyl 4-amino-1-benzyl-1H-pyrazole-3-carboxylate (compound 6)

Methyl 1-benzyl-4-nitro-1H-pyrazole-3-carboxylate (Compound 5, 3 g, 11.49 mmol, 1 eq) was dissolved in MeOH (50 mL). 5% Pd/C (300 mg, 0.1% w/w) was added to the solution. The hydrogenation reaction was performed on the mixture for 3 hours in a Parr apparatus at 30 psi conditions. Afterwards, the catalyst was removed, the solvent was removed under reduced pressure, and the residue was obtained. Impurities were removed from the residue using gel chromatography (using 50% hexane solvent of EtOAc), and solid methyl 4-amino-1-benzyl-1H-pyrazole-3-carboxylate was obtained.

1-5. Methyl 1-benzyl-4-(2-(tritylthio)acetamido)-1H-pyrazole-3-carboxylate (methyl 1-benzyl-4-(2-(tritylthio)acetamido)-1H-pyrazole- 3-carboxylate) (Compound 7) synthesis

2-(tritylthio)acetic acid (Compound 2 ) (1.73 g, 5.19 mmol, 1.2 eq), EDCI (N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride, 2.06 g, 10.82 mmol, 2.5 eq), HOBt (Hydroxybenzotriazole, 1.46 g, 10.82 mmol, 2.5 eq) , DIPEA (N,N-Diisopropylethylamine, 1.68 g (2.3 mL), 12.99 mmol, 3 eq) was added to 10mL CH ₂ Cl ₂ and stirred for 15 minutes. Methyl 4-amino-1-benzyl-1H-pyrazole-3-carboxylate (Compound 6, 1 g, 4.33 mmol, 1 eq) was added to the solution and stirred at room temperature for 12 hours. The reaction solution was diluted with CH ₂ Cl ₂ and washed with brine. The organic layer was recovered, dried over sodium sulfate, and concentrated under reduced pressure to obtain a residue. Impurities were removed from the residue using gel chromatography (using 50% hexane solvent of EtOAc), and solid methyl 1-benzyl-4-(2-(tritylthio)acetamido)-1H-pyrazole-3 was obtained. -Carboxylate (1.33 g, 56.35 %) was obtained.

1-6. Methyl 1-benzyl-4-(2-mercaptoacetamido)-1H-pyrazole-3-carboxylate (methyl 1-benzyl-4-(2-mercaptoacetamido)-1H-pyrazole-3-carboxylate) (compound 8) Synthesis

Methyl 1-benzyl-4-(2-(tritylthio)acetamido)-1H-pyrazole-3-carboxylate (Compound 7, 1.9 g, 3.47 mmol, 1 eq) was added to methylene chloride (20 mL) under argon atmosphere. ) and cooled to 0 ⁰ C in an ice bath. Trifluoroacetic acid (5 mL) was added to the colorless solution. Afterwards, triethylsilane (2.5 mL) was quickly added to the reaction solution. The reaction was monitored by TLC and was terminated after 30 minutes. The reaction was terminated by adding water, the pH was adjusted using K ₂ CO ₃ and washed with water (20 mL) and brine (20 mL). The organic layer was concentrated under reduced pressure to obtain a residue, impurities were removed from the residue using gel chromatography (using 40% hexane solvent of EtOAc), and solid methyl 1-benzyl-4-(2-mecaptoacetamido) was obtained. -1H-pyrazole-3-carboxylate (880 mg, 83.01 %) was obtained.

1-7. 1-benzyl-4-(2-mercaptoacetamido)-1H-pyrazole-3-carboxylic acid (Formula I Compound) ) synthesis

A solution of methyl 1-benzyl-4-(2-mecaptoacetamido)-1H-pyrazole-3-carboxylate (Compound 8, 870 mg, 2.85 mmol, 1 eq) in THF (10 mL) was added with NaOH (288 mg). , 5.70 mmol, 2 eq) in 3 mL of water was added and stirred for 12 hours. After removing THF and diluting with water, the pH was adjusted with HCl, extracted with CH ₂ Cl ₂ and dried over sorbium sulfate. The solvent was removed from the solution under reduced pressure to obtain yellow solid 1-benzyl-4-(2-mecaptoacetamido)-1H-pyrazole-3-carboxylic acid (770 mg, 92.77%).

The structure of the compound of formula I (1-benzyl-4-(2-mecaptoacetamido)-1H-pyrazole-3-carboxylic acid) was confirmed through proton NMR spectrum and high-resolution mass spectrometry (HRMS) spectrum.

[Formula I]

MF: C ₁₃ H ₁₃ N ₃ O ₃ S, ¹ HNMR (400 MHz, DMSO-d ₆ ) d 9.92 (s, 1H), 9.73 (s, 1H), 8.38 (s, 1H), 7.55-7.26 (m , 5H), 5.38 (s, 2H), 3.76 (s, 2H). ¹³ C NMR (100 MHz, DMSO-d ₆ ) d 165.61, 164.36, 136.63, 131.08, 128.68, 127.97, 127.87, 127.83, 123.81, 122.62, 55.76, 41.51. Mass (ESI) m/z; 314 (M+Na).

Example 2. Inhibition of Pseudomonas aeruginosa OxyR function by compounds of formula I (Figure 1)

2-1. preparation

Spotting analysis was performed to investigate the viability of serial dilutions after compound treatment. After Pseudomonas aeruginosa was grown to the logarithmic growth phase, an aliquot (3 μl) of the culture serially diluted 10-fold with LB liquid medium with or without Formula I compound (100 mM) was spotted on LB solid medium. Plates were incubated at 37°C under aerobic (18 hours) and anaerobic (32 hours) conditions.

2-2. Serial dilution growth defect experiment of Pseudomonas aeruginosa

Compounds of formula I were tested to determine whether they inhibit OxyR function. oxyR deletion mutant strains and some point mutants show a phenotype of loss of viability upon serial dilution under aerobic conditions. Compounds of formula I affected growth in serial dilution cultures under aerobic conditions of the wild type, similar to that of the o xyR deletion mutant.

The results are shown in Figure 1.

Example 3. Molecular docking model of compounds of formula I and OxyR (Figure 2)

The crystal structure of Pseudomonas aeruginosa OxyR from the Protein Data Bank (PDB code: 4Y0M), which represents the reduced structure of the regulatory domain (RD; amino acids 88–310) of OxyR, was selected as the target region, and missing hydrogens and uncharged atoms were identified. After correction, it was prepared for molecular docking studies by performing energy minimization by applying the AHARMm force until a satisfactory gradient tolerance was obtained (using Discovery Studio 2019 program).

Molecular docking analysis using the crystal structure of OxyR revealed a relatively stable interaction between the compound of formula I and OxyR RD.

The results are shown in Figure 2. In the best docked model, as shown in Figure 2A, compounds of formula I show a total of six interactions with OxyR. This is associated with participation in three hydrogen-bond interactions with Thr129, His130, and Gly197 around the pyrazole center (Figure 2b). In particular, the thiol moiety penetrated deep into the H ₂ O ₂ -binding region and formed pi-sulfur interactions with His198, which is noteworthy given that His198 is important for the reactivity of cysteine peroxide (Cys199). It should also be noted that the pyrazole group and the benzyl group participated in two pi-alkyl interactions with the thiol group of Cys199. The relatively robust packing between the Formula I compound and surrounding amino acid residues contributed significantly to the overall safety of the interaction between the Formula I compound and OxyR. These results suggest that compounds of formula I can bind to two regions (Thr129-His130 and Gly197-His198-Cys199) of OxyR RD, in a manner that mimics the phenotype of the oxyR deletion mutant strain more than that of the oxyR RD point mutant. may damage its function.

Example 4. Changes in the regulatory function of OxyR by compounds of formula I (Figure 3)

4-1. Experiment to confirm the redox state of OxyR (Figure 3a)

4-1-1. preparation

Changes in OxyR redox state were investigated using a thiol group capture method using 4-acetamido-4'-maleimidylstilbene-2,2'-disulfonic acid (AMS). First, the oxyR deletion mutant strain carrying a plasmid expressing FLAG-tagged OxyR protein was grown to logarithmic growth phase in LB broth with or without Formula I compound (200 μM) at 37°C. And after treatment with 1mM H ₂ O ₂ , 110 μl of 100% (wt/vol) trichloroacetic acid (TCA) was added to an aliquot (1 ml) of the culture. The cell pellet was resuspended in 400 μl of 10% TCA, and the cells were disrupted by sonicating. After centrifugation, the pellet was mixed with 40 μl of AMS buffer (0.5 M Tris-HCl [pH 8.0], 20 mM AMS, 2% SDS, 100 mM NaCl, 1 mM EDTA, and 5% glycerol) and prepared for cell extraction. After incubation for 1 hour in the dark, Western blot analysis was performed using anti-FLAG M2 antibody (Sigma).

4-1-2. Analysis of redox state of OxyR through electrophoresis

The potential effects of compounds of formula I on the redox cycle of OxyR were investigated under oxidative stress conditions. We previously utilized the thiol alkylation method using AMS on FLAG-tagged OxyR protein at the C terminus, and observed three oxidation states of OxyR: two species I and II, which migrate faster than reduced OxyR, and ∼50 -kDa species (III).

The results are shown in Figure 3. As shown in Figure 3a, three oxidized OxyR species were clearly observed under non-reducing conditions without 2-mercaptoethanol (β-ME) treatment, and most of the oxidized species were released after 10 min of H ₂ O ₂ exposure. decreased rapidly in vivo. Since two prominent bands were observed under reducing conditions (i.e., β-ME treatment), species I may possess three free thiols, species II and III may possess two free thiols, and both It is likely to migrate faster than the reduced species with four free thiols. Although detailed characterization of the oxidized species requires more extensive chemical confirmation based on mass spectrometry, it is clear that more than one type of disulfide bond is formed during OxyR peroxide sensing.

However, in contrast, the oxidized state of the OxyR protein in the presence of compounds of formula I appeared to be significantly different in at least three aspects: first, species III was not observed at all; Second, faster moving bands were observed; Finally, no drastic decrease was observed at 10 min. Most importantly, there is no apparent difference in the band profile between non-reducing and reducing conditions, suggesting that no disulfide bonds were formed in the presence of compounds of formula I during H ₂ O ₂ treatment. These results and the proposed direct interaction between the Formula I compound and the OxyR RD suggest that the Formula I compound indeed binds OxyR and that ligand binding disrupts the formation of a disulfide bond involving the OxyR RD, which, by an unknown mechanism, inhibits OxyR function. You can see that it can cause damage.

4-2. Expression pattern analysis experiment of OxyR regulated genes (Figure 3b)

4-2-1. preparation

To measure katA transcriptional activity, the katA promoter- lacZ (β-galactosidase) transcriptional fusion plasmid (based on pQF50) was introduced into Pseudomonas aeruginosa by electroporation, and the culture was grown to mid-logarithmic (OD ₆₀₀ of 0.3) growth phase. β-galactosidase activity was measured.

4-2-2. using a reporter katA Analysis of transcriptional regulation patterns of genes

Given that OxyR acts as both an activator and repressor for the katA gene, the katA promoter -lacZ transcription fusion was used to determine the effect of compounds of formula I on OxyR function. As shown in Figure 3b, transcription of katA was partially reduced in the presence of compound I, which is consistent with the observation that treatment with compound I resulted in the phenotype of the oxyR deletion mutant strain (Figure 1).

Point mutants for peroxidized and degraded cysteines were found to behave either as a persistently active form (Cys199 mutation) or as an unresponsive (i.e. locked) inhibitory form (Cys208 mutation). It should be noted that compounds of formula I target OxyR RD in a way that affects the redox cycle of OxyR, but may reduce OxyR function (Figures 1 and 3). This needs to be addressed systematically, not only with regard to modifications of cysteine residues, but also with regard to the function of the amino acid residues (Thr129, His130, Gly197, His198 and Cys199) that are believed to interact with compounds of formula I and the function of OxyR that recognizes peroxides. Greater clarity can be achieved through a more precise understanding of the complex molecular details in the redox cycle.

Example 5. Selective antibacterial efficacy of compounds of formula I (Figure 4)

5-1. preparation

Drosophila systemic infection was performed to evaluate antibacterial efficacy. Drosophila melanogaster Oregon R fruit flies were grown on cornmeal-dextrose medium [0.93% agar, 6.24% dry yeast, 4.08% cornmeal, 8.62% dextrose, 0.1% methyl paraben, and 0.45% ( v/v) propionic acid] was used to grow and maintain. PBS-diluted bacterial suspensions containing Pseudomonas aeruginosa PA 14 (10 ⁷ cfu/ml) or Staphylococcus aureus SA3 (10 ⁸ cfu/ml) grown to an OD ₆₀₀ of 3.0 in the presence or absence of Formula I compound (200 μM). 4-5 day old adult female flies were systemically infected through the ventral thorax using a 0.4 mm needle. Survival of infected flies was monitored for up to 48 hours after infection. Flies that died within 12 hours were excluded from mortality determinations. Mortality assays were repeated at least three times.

5-2. Experiment on the antibacterial efficacy of Pseudomonas aeruginosa using a Drosophila infection model

Since OxyR is required for virulence of Pseudomonas aeruginosa, compounds of formula I may affect the virulence properties of Pseudomonas aeruginosa. To test this possibility, we used a Drosophila systemic infection model to evaluate the antibacterial efficacy of Formula I compounds, as utilized in previous studies. Considering the solubility of the compounds in Drosophila rearing medium, bacterial cells pretreated with compounds of formula I were pricked and infected as described in the methods section.

The results are shown in Figure 4. As shown in Figure 4A, a reduction in toxicity was clearly observed by treatment with compounds of formula I on flies infected with Pseudomonas aeruginosa under the experimental conditions of the present invention.

The present invention also noted that treatment with Formula I compounds did not show antibacterial efficacy against methacillin-resistant Staphylococcus aureus SA3 (Figure 4b). SA3 or other Gram-positive pathogenic bacteria have a Fur paralog, PerR, as a peroxide sensor, while OxyR is widely found in Gram-negative bacteria. Antibacterial efficacy against SA3 was not confirmed under any of the experimental infection conditions tested by the present invention, and because the compound of formula I specifically impaired OxyR function, it was used against Gram-negative pathogenic bacteria (e.g., Pseudomonas aeruginosa) that have OxyR as a master peroxide sensor. It can be concluded that it specifically targets ).

Claims (5)

A pyrazole-based compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof:
[Formula I]

The method according to claim 1, wherein the pharmaceutically acceptable salt is a pyrazole-based compound or a pharmaceutically acceptable salt thereof, characterized in that it is a salt with a metal ion, ammonia, or an organic base. An anti-toxin agent containing a pyrazole-based compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof:
[Formula I]
The anti-virulence agent according to claim 3, which exhibits selective activity against Gram-negative bacterial pathogens having OxyR, an active oxygen sensor.
The method of claim 3, wherein the anti-venom agent against Pseudomonas aeruginosa