Application of diphenyl butyl piperidine derivative in preparation of quorum effect inhibitor

Abstract

The invention relates to an application of pimozide in preparation of a population effect inhibitor. The invention verifies that pimozide can inhibit the population effect of bacteria through a violaceum violaceum luminescence method, and further proves that pimozide can inhibit the population effect of pseudomonas aeruginosa and the generation of virulence factors through the inhibition of biological back membrane formation and the inhibition effect of virulence factor generation. The compound is expected to be developed into a clinically available group effect inhibitor, and has an important research value for solving the problems of bacterial infection and drug resistance.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical chemical industry, specifically relating to the application of pimozide in the preparation of quorum effect inhibitors. Background Technology

[0002] Bacteria communicate using small, self-generated signaling molecules called auto-inducers (AIs). During bacterial growth, these AIs are continuously produced and secreted into the surrounding environment. When the concentration of these signaling molecules reaches a certain threshold, they trigger the expression of relevant genes within the bacteria to adapt to environmental changes. This regulatory system is known as the bacterial quorum sensing signaling system, or QS system for short. QS enables single-celled bacteria to mimic multicellular organisms, performing behaviors that they cannot do as single-celled individuals.

[0003] Bacterial quorum sensing enables bacteria to regulate the expression of certain genes within a population, such as the regulation of antibiotic production, bioluminescence, nitrogen fixation gene regulation, Ti plasmid conjugation and transfer, expression of toxic genes, pigment production, bacterial swarming, and biofilm formation. In the late 1970s, scientists discovered that natural or synthetic quorum sensing modulators (including agonists and inhibitors) can interfere with signal transduction systems and regulate the expression of harmful bacterial genes.

[0004] In 1970, Nealson et al. first discovered quorum sensing in *Vibrio fischeri*, where the bacteria produce bioluminescence when the colony density reaches a high level. Further research revealed that in *V. fischeri*, N-acetyl homoserine lactones (AHL), synthesized by the LuxI protein, activate the lux operon by interacting with the transcription activator luxR. Similar regulatory systems have been found in many Gram-negative and Gram-positive bacteria. Their mechanism of action is that when bacterial colony density is low, the inducin synthase gene produces a basal level of expression, causing a small amount of self-induced signals. These signals diffuse extracellularly and are immediately diluted in the surrounding environment. However, as bacterial colony density increases and the concentration reaches a threshold, they infiltrate the cell and bind to transcriptional regulatory proteins, forming a polymer of transcriptional regulatory protein signaling molecules. This polymer can bind to a specific DNA sequence of chromosomal signaling molecules, leading to the expression of target genes, including the signaling molecule synthesis gene, and the production of more signaling molecules. Although the concept of this kind of information exchange between bacteria has been proposed for some time, systematic research has mainly focused on the last decade. This effect has now been confirmed to exist in a variety of bacteria. For example, *Chromobacterium violaceum* and *Vibrio fischeri* share the same mechanism, producing C6-HSL as a self-inducing molecule, with its receptor protein being CviR.

[0005] Pseudomonas aeruginosa (PA) is a significant opportunistic pathogen causing hospital-acquired infections, widely distributed in nature (soil, air, and water). After infecting humans, PA causes disease by secreting various virulence factors and forming a biofilm that adheres to the patient's body, damaging human tissues, inducing inflammation, and impairing the host's immune system. PA possesses multiple antibiotic resistance mechanisms, with the QS-regulated biofilm being a key component. PA's quorum effect is primarily driven by the LasI / LasR system, a quorum effect regulatory mechanism widely present in Gram-negative bacteria. Inhibiting the quorum effect of Pseudomonas aeruginosa also has a broad-spectrum inhibitory effect on Gram-negative bacterial infections. Quorum sensing modulators do not interfere with the normal physiological functions of cells in the body, and are therefore considered a new direction in the development of antibacterial drugs. Quorum sensing inhibitors can be used in combination with antibiotics to increase the sensitivity of pathogenic bacteria to antibiotics, or they can be used alone to maintain the non-toxic state of pathogens, especially opportunistic pathogens. Treatment includes, but is not limited to, diseases caused by Gram-negative bacteria such as Escherichia coli, Proteus, Shigella, Klebsiella pneumoniae, Brucella, Haemophilus influenzae, Haemophilus parainfluenzae, Moraxella catarrhalis, Acinetobacter spp., Yersinia spp., Legionella pneumophila, Bordetella pertussis, Bordetella parapertussis, Shigella spp., Pasteurella spp., Vibrio cholerae, and Bacillus parahaemolyticus, including but not limited to peritonitis, cholecystitis, cystitis, diarrhea, endocarditis, gastroenteritis, empyema, and sepsis. They are particularly effective in treating diseases caused by drug-resistant Gram-negative bacteria that are insensitive to existing antibiotics.

[0006] This invention discovers a marketed drug with quorum sensing modifiers that can be used to treat diseases caused by Gram-negative bacteria, especially those caused by drug-resistant Gram-negative bacteria. Summary of the Invention

[0007] This invention provides the pharmaceutical use of the compound pimozide (CAS 2062-78-4) in the preparation of quorum effect inhibitors, and discloses the activity of pimozide in inhibiting the quorum effect of violaceum bacillus and Pseudomonas aeruginosa.

[0008]

[0009] This invention verifies that pimozide can inhibit the quorum effects of *Pseudomonas aeruginosa* and *C. aeruginosa* through luminescence inhibition experiments and biofilm formation inhibition experiments. Furthermore, quantitative analysis of virulence factors demonstrates that pimozide can inhibit the production of virulence factors by *P. aeruginosa*, thereby reducing its pathogenicity and achieving treatment for infections caused by Gram-negative bacteria represented by *P. aeruginosa*.

[0010] The positive effects of this invention are as follows:

[0011] Pimozide is a diphenylbutylpiperidine derivative dopamine D2 antagonist that also significantly inhibits the STAT3 / STAT5 pathway. Marketed in 1969, it is primarily used clinically to treat schizophrenia, paranoia and hypochondria, Tourette syndrome, etc., and can also inhibit the proliferation of various cancer cells, such as colorectal cancer, human osteosarcoma, and hepatocellular carcinoma. No publicly available research reports have documented its antibacterial effects or its ability to inhibit bacterial quorum effects.

[0012] This invention verifies that pimozide can inhibit the quorum effects of *Pseudomonas aeruginosa* and *C. aeruginosa* through luminescence inhibition experiments and biofilm formation inhibition experiments. Further quantitative analysis of virulence factors demonstrates that pimozide can inhibit the production of virulence factors by *P. aeruginosa*, thereby reducing its pathogenicity and achieving treatment for infections caused by Gram-negative bacteria represented by *P. aeruginosa*. Because it only inhibits the production of pathogenic factors such as virulence factors and does not inhibit normal bacterial growth, it is less likely to induce drug resistance, and has further research and application value for solving the problem of classical antibiotic resistance. Detailed Implementation

[0013] Example 1: Activity evaluation of Chromobacterium violaceum quorum sensing regulator

[0014] 1. Initial screening method

[0015] 1.1 Preparation: Weigh pimozide (purchased from MedChemExpress, HY-12987) and dissolve it in 200 μl of DMSO. Weigh 5.0 mg of the inducer N-hexanoylhomoserine lactone C6-HSL (original concentration 0.125M, purchased from Sigma) and dissolve it in 200 μl of DMSO. Both compounds and inducers were stored at 4℃ for later use. Chlorella vulgaris was cultured in a shaker at 30℃ and 200 rpm.

[0016] 1.2 Screening method for compounds with inhibitory activity: The inducer C6-HSL was serially diluted 1000 times. Then, 15 μl and 400 μl of logarithmic phase Chlorobacterium violaceum were mixed evenly and added to 5 ml of melted semi-solid LB medium. The mixture of inducer, Chlorobacterium violaceum and semi-solid LB was then poured onto a solid LB plate. After the plate solidified, a grid was drawn. 1 μl of the dissolved test compound was spotted onto the plate. After the compound on the plate dried, it was placed in an oven at 30℃ and inverted for 16-18 h.

[0017] IC50 determination of compounds that inhibit quorum sensing of *Synthobacter violaceus*

[0018] 2.1 Take a 12-well plate and label each well from left to right and top to bottom as the original concentration, 2 times, 4 times, 8 times, 16 times, 32 times, 64 times, 128 times, 256 times dilute, DMSO group, and blank group.

[0019] 2.2 Select a single colony of indicator bacteria (Chroma violaceum CV026) grown on an LB solid plate and culture it in 5 ml of fresh LB liquid medium until the logarithmic growth phase. Then, take 50 μl of the colony and inoculate it into 5 ml of LB medium. Culture it until the OD is about 1.0. Then mix it with LB medium at a ratio of 1:9 (the OD is about 0.15 at this time). Then add 2 ml of the mixture to each well of a 12-well plate.

[0020] 2.3 After dissolving the compound DMSO with quorum inhibition activity, 10 μL of each compound was added to 10 μL of DMSO solution to achieve a 2-fold dilution. This process was repeated to dilute each compound to the highest factor of 256.

[0021] 2.4 Add 15 μL of inducer C6-HSL diluted 1000 times (original concentration 0.125 M, purchased from Sigma) and 8 μL of each dilution gradient of the compound to each well. Add 8 μL of DMSO to the DMSO control group and 8 μL of LB medium to the blank control group. Finally, ensure that the 2 ml of culture in the 12-well plate is thoroughly mixed.

[0022] 2.5 Place the 12-well culture plate in a shaker at 30℃ and incubate at 130 rpm for 10-12 hours.

[0023] 2.6 After the culture is completed, take 1 ml of culture from each well into a 1.5 ml EP tube, centrifuge at 12000 rpm for 10 minutes, remove the culture supernatant, add 500 μl of DMSO to each EP tube to dissolve the purple pigment, and centrifuge again at 12000 rpm for 10 minutes after the pigment is completely dissolved. Then take 200 μl of the upper pigment into a 96-well culture plate and measure its absorbance at 585 nm using a microplate reader. Plot the relationship between the absorbance and the corresponding concentration to obtain the IC50 value.

[0024] The results of the initial screening for activity are shown in the table below:

[0025] Table 1. Initial screening activity and IC50 of the compounds inhibiting CV026 quorum sensing.

[0026] compound Primary screening activity <![CDATA[IC 50 (uM)]]> Pimozzi +++ 0.387±0.008

[0027] In Table 1, “+”, “++”, and “+++” indicate that the compounds have quorum-sensing activity, and the more “+” signs there are, the higher the activity.

[0028] Example 2: Activity evaluation of a quorum sensing regulator against Pseudomonas aeruginosa

[0029] 2.1 Experimental Materials

[0030] 2.1.1 Experimental bacterial strains

[0031] In this experiment, the Pseudomonas aeruginosa PAO1 strain used was derived from a surviving strain in our laboratory.

[0032] 2.2 Experimental Equipment

[0033] 2.2.1 Preparation of experimental solutions

[0034] (1) Preparation method of LB nutrient broth medium (1L):

[0035] First, weigh out 10g of tryptone, 10g of NaCl, and 5g of yeast extract, and add them to a 1000ml beaker. Add 700mL of distilled water to dissolve them. Adjust the pH to 7.2, then bring the volume to 1L. For solid LB culture, add an additional 15g of agar.

[0036] (2) Preparation of PB medium (1L):

[0037] First, weigh out 20g of peptone, 10g of K2SO4, and 3g of MgCl2, then dissolve and dilute them in 1L of distilled water until the volume is brought to a final volume. Finally, autoclave the mixture and store it at 4°C.

[0038] (3) Preparation of PTSB medium (1L):

[0039] First, weigh out 50g of peptone, 17g of tryptone, 5g of NaCl, 2.5g of K2HPO4, 2.5g of glucose, and 3g of soybean peptone. Then, dissolve and bring the contents to a final volume with 1L of distilled water, autoclave, and store at 4℃.

[0040] (4) Preparation of PPGAS medium (1L):

[0041] Weigh out 20 mM KCl, 20 mM NH4Cl, 0.5% glucose, 1.6 mM MgSO4·7H2O, 120 mM Tris HCl, and 1.0% peptone sequentially. Dissolve and bring to volume with 1 L of distilled water, adjust pH to 7.2, and finally autoclave and store at 4°C.

[0042] (5) Preparation of TSB liquid culture medium (1L):

[0043] First, weigh out 5g of soybean peptone, 15g of tryptone, and 5g of NaCl, then dissolve and dilute them in 1000mL of distilled water, adjust the pH to 7.2±0.2, and autoclave for later use.

[0044] (6) Preparation of PGS solid culture medium (1L):

[0045] First, weigh out peptone (10g), glucose (10g), sorbitol (27.315g), NaCl (10g) and agar (10g), then dissolve and bring to a final volume in 1000mL of distilled water, and autoclave for later use.

[0046] (7) Preparation of 1M potassium phosphate buffer (100mL):

[0047] Method for preparing mother liquor:

[0048] Preparation of 1M KH2PO4: Weigh out 13.609g of KH2PO4 and dissolve and dilute to volume with 100mL of distilled water.

[0049] Preparation of 1M K2HPO4: Weigh out 22.822g of K2HPO4·3H2O, then dissolve and dilute with 100mL of distilled water.

[0050] Preparation of 1M K2HPO4 / KH2PO4 (pH=7.0): Measure 61.5 mL of 1M K2HPO4 solution and 38.5 mL of 1M KH2PO4 solution respectively, then stir and mix them evenly, and then autoclave for later use.

[0051] (8) Preparation of 0.2M PBS buffer (100mL):

[0052] Preparation of 0.2M Na2HPO4: First, weigh 7.16g of Na2HPO4·12H2O, then dissolve and dilute to volume with 100mL of distilled water.

[0053] Preparation of 0.2M NaH2PO4: First, weigh 3.12g of NaH2PO4·2H2O, then dissolve and dilute to volume with 100mL of distilled water.

[0054] Preparation of 0.2M PBS (pH=7.4): Measure 19 mL of 0.2M NaH2PO4 solution and 81 mL of 0.2M Na2HPO4 solution respectively, mix them thoroughly, and then autoclave for later use.

[0055] (9) Preparation method of 0.1% crystal violet staining solution:

[0056] Weigh 0.1g of solid crystal violet powder, add 100mL of distilled water to dissolve it, and then make up to a final volume.

[0057] (10) Preparation method of swarming medium (1L):

[0058] Weigh out 8g of nutrient broth (BD), 3g of agar (BD), and 5g of glucose, add them to 1000mL of distilled water, dissolve them, and then make up to a final volume.

[0059] (11) Preparation method of substrate elastin-Congo red (ECR) reaction buffer:

[0060] First, dilute 1M Tris-HCl (pH=7.2) with ddH2O to a concentration of 0.1M. Then, add CaCl2 to the diluted solution to bring the final concentration to 1mM. Finally, add elastin-Congo red to bring the final concentration to 20mg / mL. Store at 4℃ for later use.

[0061] (12) Preparation method of 1M Tris-HCl:

[0062] First, weigh 121.1g of Tris, then dissolve it in ddH2O, adjust the pH to 7.2 with concentrated HCl (7.6mL), and finally bring the volume to 100mL with ddH2O. Autoclave the solution and store it at 4°C.

[0063] (13) Preparation method of phenol-concentrated sulfuric acid reagent:

[0064] First, take 139 mL of 95% concentrated sulfuric acid and slowly add it along the wall of the bottle to 111 mL of distilled water while stirring. After the solution cools to 25°C, add 0.475 g of phenol and stir until it is thoroughly mixed.

[0065] (14) Preparation method of azocasein-Tris-HCl (pH=7.8) solution:

[0066] First, dissolve the azo casein in 50mM Tris-HCl solution (1M Tris-HCl diluted 20 times) to prepare a 2% (w / v) solution.

[0067] (15) Preparation of experimental reagents:

[0068] Preparation of compound solution: The compound to be tested was prepared to a concentration of 65 mM using dimethyl sulfoxide (DMSO), and then stored at -20°C for later use.

[0069] Preparation of antibiotic solution: Prepare a 6.4 mg / mL solution of tobramycin (purchased from Simga) with distilled water and store at -20°C for later use.

[0070] 2.3 Experimental Methods

[0071] 2.3.1 Quantitative Analysis of Biofilms

[0072] First, *Pseudomonas aeruginosa* PAO1 was inoculated into LB medium and then cultured to the logarithmic growth phase at 37°C and 150 rpm. After successful culture, the bacterial culture was diluted to OD using fresh LB medium. 600 =0.05. After the bacterial suspension was diluted, 98 μL of the bacterial suspension and 2 μL of different concentrations of the test compounds were added to each well of a 96-well plate, resulting in concentrations of 325 μM, 162.5 μM, 81.25 μM, 40 μM, 20 μM, 10 μM, 5 μM, and 2.5 μM, respectively. Then, 100 μL of the bacterial suspension was added to the control group. Each well was divided into three replicates, and the plate was incubated at 37 °C for 20 h. The bacterial suspension was then aspirated, and the biofilm was slowly washed twice with ddH2O. The washed biofilm was then air-dried in a fume hood. Next, 150 μL of 0.1% crystal violet solution was added to each well for staining. After staining for 10 min, the plate was washed three times with ddH2O, while simultaneously removing the crystal violet from each well with 150 μL of 33% glacial acetic acid. Finally, the plate was transferred to a new 96-well plate to measure OD. 595 Biofilm inhibition rate = OD 595 Blank-OD 595 (sample) / OD 595 100% blank.

[0073] 2.3.2 Analysis of the compound's activity on PAO1 growth

[0074] Dilute P. aeruginosa PAO1 in the logarithmic growth phase with fresh LB medium to OD. 600 =0.05. 98 μL of diluted bacterial culture and 2 μL of the test compound solution were added to each well of a 96-well plate, resulting in final compound concentrations of 162.50 μM, 81.25 μM, 40.00 μM, 20 μM, 10 μM, 5 μM, and 2.5 μM in each well, respectively. Wells without added compound served as blank controls. The plate was incubated at 37°C for 24 h, and OD values ​​were measured at 0 h, 2 h, 4 h, 6 h, 8 h, 10 h, 12 h, and 24 h. 600 .

[0075] 2.3.3 Quantitative analysis of virulence factors

[0076] (1) Quantitative analysis of Pseudomonas aeruginosa:

[0077] First, *Pseudomonas aeruginosa* PAO1 cultured in LB medium was cultured to the logarithmic development phase at 37℃ and 150 rpm, and then diluted 10-fold with fresh PB medium. Next, 4.95 mL of bacterial culture and 50 μL of the test compound were added to a 50 mL Erlenmeyer flask, resulting in final concentrations of 325.0 μM, 162.5 μM, 81.25 μM, 20 μM, 10 μM, 5 μM, and 2.5 μM, respectively. After thorough mixing, the mixture was cultured at 37℃ and 150 rpm for 16 h. After 16 h, the cultured reaction mixture was centrifuged at 5000 rpm for 10 min, and the supernatant was collected. 3 mL of chloroform was added to the supernatant for extraction. The extracted organic layer was then reacted with 1 mL of 0.2 N HCl. The organic layer was centrifuged again at 5000 rpm for 10 min, the upper layer was collected and added to a 96-well plate, and the OD was measured. 520 Inhibition rate = (OD) 520 Blank-OD 520 (sample) / OD 520 100% blank.

[0078] (2) Quantitative analysis of rhamnolipin:

[0079] a. Rhamnose lipid content - OD 421 Standard curve creation:

[0080] First, 0.01 g of rhamnose was prepared into a 1 mg / mL rhamnose standard stock solution using 10 mL of distilled water. Then, the stock solution was successively diluted to prepare different concentrations of 50 μg / mL, 100 μg / mL, 200 μg / mL, and 400 μg / mL, with distilled water serving as the control group. 100 μL of each concentration of stock solution and distilled water was taken and added to centrifuge tubes containing 900 μL of phenol-concentrated sulfuric acid reagent. After heating in an 80°C water bath for 30 min, the tubes were removed, cooled, and their OD values ​​were measured. 421 Plot the rhamnose content-OD 421 Standard curve.

[0081] b. Quantitative analysis of rhamnolipin:

[0082] First, *Pseudomonas aeruginosa* PAO1 cultured in LB medium was cultured to the logarithmic growth phase at 37°C and 150 rpm. Then, the bacterial culture was diluted to OD using PPGAS medium. 600=0.05. Add 3 mL of cultured bacterial solution and the test compound to each conical flask to achieve final compound concentrations of 325.0 μM, 162.5 μM, 81.25 μM, 20 μM, 10 μM, 5 μM, and 2.5 μM. Flasks without added compound serve as blanks. Incubate the prepared compounds at 37 °C and 150 rpm for 48 h. After 48 h of incubation, remove the bacterial solution and centrifuge at 5000 rpm for 10 min. Collect the supernatant and adjust the pH to 2 with concentrated hydrochloric acid. Take 1 mL of the supernatant and extract twice with 1 mL of ethyl acetate. Place the extract in a fume hood and wait for the ethyl acetate to evaporate. Add 500 μL of distilled water to dissolve the rhamnose, and take 100 μL of this solution. Add it to 900 μL of a phenol-sulfuric acid reagent mixture, and then heat in a water bath at 80 °C for 30 min. After 30 min, remove the solution and allow it to cool for 15 min before measuring the OD. 421 The data were substituted into the standard curve to determine the rhamnose content in the sample, and finally the rhamnose lipid content in the sample was calculated. The inhibition rate was then calculated by comparing the results with the blank control.

[0083] Conversion formula: 1 μg rhamnose corresponds to 2.5 μg rhamnolipid in the sample.

[0084] (3) Quantitative analysis of elastase:

[0085] The logarithmic phase PAO1 was diluted and added to the compound to a final concentration of 325.0 μM, 162.50 μM, 81.25 μM, 40.00 μM, 20.00 μM, 10 μM, 5 μM, and 2.5 μM. The mixture was incubated at 37°C for 24 h at 150 rpm. The mixture was then sterilized by filtration through a 0.22 μm filter, and the elastase activity was measured according to the EnzCheck™ Elastase Assay Kit method.

[0086] 2.4. Experimental Results

[0087] The results of the activity evaluation are shown in the table below:

[0088] Table 2. Inhibitory effect of pimozide on biofilm formation in Pseudomonas aeruginosa PAO1

[0089] compound concentration 325.0μM 162.5μM 81.25μM 40.0μM Inhibition rate (%) 85.5 82.2 75.5 70.4 compound concentration 20.0μM 10.0μM 5.0μM 2.5μM Inhibition rate (%) 65.8 55.3 42.5 23.3

[0090] Table 3 Effects of pimozide on PAO1 growth activity

[0091] compound concentration 162.5μM 81.25μM 40.0μM 20.0μM Growth rate (%) 99.4 100.5 99.7 101.2 compound concentration 10.0μM 5.0μM 2.5μM blank Growth rate (%) 99.8 100.3 102.1 100

[0092] Table 4. Effects of pimozide on Pyocyanin production in PAO1

[0093] compound concentration 325.0μM 162.5μM 81.25μM 40.0μM Inhibition rate (%) 95.5 92.2 82.5 75.4 compound concentration 20.0μM 10.0μM 5.0μM 2.5μM Inhibition rate (%) 68.8 60.3 55.5 43.3

[0094] Table 5. Effects of pimozide on rhamnolipid formation in PAO1.

[0095] compound concentration 325.0μM 162.5μM 81.25μM 40.0μM Inhibition rate (%) 96.8 96.2 80.5 72.4 compound concentration 20.0μM 10.0μM 5.0μM 2.5μM Inhibition rate (%) 64.8 58.3 45.5 38.3

[0096] Table 6. Effects of pimozide on elastase production in PAO1.

[0097] compound concentration 325.0μM 162.5μM 81.25μM 40.0μM Inhibition rate (%) 85.1 76.5 69.1 59.4 compound concentration 20.0μM 10.0μM 5.0μM 2.5μM Inhibition rate (%) 52.1 44.3 35.5 28.5

[0098] The results of activity evaluation showed that pimozide could significantly inhibit the quorum effect of *Pseudomonas aeruginosa*, with an IC50 of 0.387 ± 0.008 μM. Moreover, it could effectively inhibit the formation of the biofilm and the production of virulence factors of *Pseudomonas aeruginosa* PAO1 (IC50 approximately 5.0–10.0 μM), thereby reducing the pathogenicity of *Pseudomonas aeruginosa* PAO1. At the same time, it had no effect on the growth of *Pseudomonas aeruginosa* PAO1. This antibacterial infection effect that does not interfere with the normal physiological functions of cells in vivo is considered a new direction for the development of antibacterial drugs. It can be used in combination with antibiotics to increase the sensitivity of pathogenic bacteria to antibiotics, or it can be used alone to maintain the non-toxic state of pathogens, especially opportunistic pathogens. Treatment includes, but is not limited to, diseases caused by Gram-negative bacteria such as Escherichia coli, Proteus, Shigella, Klebsiella pneumoniae, Brucella, Haemophilus influenzae, Haemophilus parainfluenzae, Moraxella catarrhalis, Acinetobacter spp., Yersinia spp., Legionella pneumophila, Bordetella pertussis, Bordetella parapertussis, Shigella spp., Pasteurella spp., Vibrio cholerae, and Bacillus parahaemolyticus, including but not limited to peritonitis, cholecystitis, cystitis, diarrhea, endocarditis, gastroenteritis, empyema, and sepsis. It is particularly effective in treating diseases caused by drug-resistant Gram-negative bacteria that are not sensitive to existing antibiotics.

Claims

1. Use of pimozide or a pharmaceutically acceptable salt thereof in the preparation of an inhibitor of the group effect. The pimozide is represented by Formula I:

2. Use of pimozide for the preparation of a group effect inhibitor according to claim 1, characterized in that The pimozide is a pharmaceutically acceptable carrier.

3. Use of pimozide for the preparation of a group effect inhibitor according to claim 1, characterized in that The use is the use of pimozide in the preparation of a medicament for the treatment of an infectious disease.

4. Use of pimozide according to claim 1 for the preparation of a group effect inhibitor, characterized in that The infectious disease refers to an infection caused by bacteria.

5. The use of pimozide according to claim 1 or 3 for the preparation of a group effect inhibitor, characterized in that The bacterial infection refers to an infection caused by gram-negative bacteria such as Pseudomonas aeruginosa.

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