Aryl dicarboxylic acid compound and antibacterial application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to an aryl dicarboxylic acid compound, a preparation method and application of the aryl dicarboxylic acid compound as an antibacterial agent. The compound is shown as a formula (1), wherein R1, R2, R3, R4, R5, R6, R7, R8, R9 and R10 are independently selected from hydrogen and halogen. Two ends of the molecule are connected through an N-phenylpropanamide connecting arm, the molecule is a compound with a novel structure, and the series of compounds have a relatively strong inhibition effect on acinetobacter baumannii and are worthy of further research and development.

Description

Technical Field

[0001] This invention belongs to the field of antibacterial drug technology, specifically relating to aryl dicarboxylic acid compounds, their preparation methods, and their uses as antibacterial drugs. Background Technology

[0002] In recent years, the emergence of drug-resistant bacteria and pathogens such as mycoplasma has led to a gradual decrease in the availability of effective antibiotics. On the other hand, antimicrobial drug development has been relatively slow: the number of approved antibiotics is small, and most are still of known structural types, making them prone to cross-resistance. Statistics show that in the past 20 years, only three new structural types of antimicrobial drugs have been approved globally, and these only target Gram-positive bacteria such as Staphylococcus aureus, without any new structural types targeting Gram-negative bacteria such as Acinetobacter baumannii. Therefore, antimicrobial drugs with new structural types or new mechanisms of action address significant clinical needs.

[0003] The compound involved in this patent belongs to the aryl dicarboxylic acid class of compounds. Due to the N-phenylpropionamide linking arm connecting both ends of the molecule, it is a novel compound structure. This compound exhibits strong antibacterial activity against Acinetobacter baumannii, with a minimum inhibitory concentration (MIC) of 0.5 μg / mL against the standard strain ATCC19606. This activity value is close to that of the clinical drug levofloxacin (MIC: 0.25 μg / mL). Therefore, the compound involved in this patent has the potential for further development into a clinical antibacterial drug. Summary of the Invention

[0004] The technical problem addressed by this patent is to overcome the shortcomings of existing antimicrobial drugs for drug-resistant bacteria by providing compounds with novel structures or their pharmaceutically acceptable salts, which exhibit strong antibacterial activity. This patent provides aryl dicarboxylic acid compounds, their preparation methods, and their uses as antimicrobial drugs.

[0005] To solve the technical problem of this invention, the present invention provides the following technical solution:

[0006] In the first aspect, an aryl dicarboxylic acid compound is provided, which is a compound as shown in formula (1):

[0007]

[0008] Among them: R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 Independently selected from hydrogen and halogens;

[0009] The halogen is selected from fluorine, chlorine, bromine, and iodine.

[0010] Preferably, the compound provided by the present invention is:

[0011] 5-[3-(3-{3-carboxy-4-[5-(4-chlorophenyl)thiophene-2-sulfonamido]phenyl}propamido)phenyl]-2-[5-(4-chlorophenyl)thiophene-2-sulfonamido]benzoic acid:

[0012]

[0013] Secondly, the present invention provides the antibacterial use of the above-mentioned aryl dicarboxylic acid compounds or their use in the preparation of antibacterial drugs.

[0014] Preferably, the antibacterial use is for the treatment and prevention of infectious diseases in humans or animals caused by pathogenic microorganisms such as bacteria, mycoplasma, chlamydia, rickettsia, spirochetes, and fungi.

[0015] More preferably, the bacteria are Acinetobacter baumannii, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Enterococcus faecium, Clostridium difficile, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, etc.; the mycoplasma are Mycoplasma pneumoniae, Ureaplasma urealyticum, Mycoplasma hominis, Mycoplasma genitalium, etc.; and the chlamydia are Chlamydia pneumoniae, Chlamydia psittaci, Chlamydia trachomatis, and Chlamydia bovis, etc.

[0016] More preferably, the Acinetobacter baumannii includes drug-sensitive Acinetobacter baumannii, multidrug-resistant Acinetobacter baumannii, pan-drug-resistant Acinetobacter baumannii, carbapenem-resistant Acinetobacter baumannii, and pan-drug-resistant Acinetobacter baumannii; the Staphylococcus aureus includes methicillin-sensitive Staphylococcus aureus and methicillin-resistant Staphylococcus aureus; the Staphylococcus epidermidis includes methicillin-sensitive Staphylococcus epidermidis and methicillin-resistant Staphylococcus epidermidis; the Enterococcus faecalis includes vancomycin-sensitive Enterococcus faecalis and vancomycin-resistant Enterococcus faecalis; and the Enterococcus faecium includes vancomycin-sensitive Enterococcus faecium and vancomycin-resistant Enterococcus faecium; and the Mycoplasma is selected from drug-sensitive Mycoplasma pneumoniae or drug-resistant Mycoplasma pneumoniae.

[0017] Thirdly, the present invention provides an antibacterial pharmaceutical composition comprising the above-mentioned aryl dicarboxylic acid compound or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers.

[0018] Furthermore, the carrier includes excipients, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorbents, lubricants, etc., commonly used in the pharmaceutical field.

[0019] Preferably, the drug is administered in the form of an injection, tablet, pill, capsule, suspension, emulsion, or ointment, and the route of administration is selected from intravenous or intramuscular injection, oral administration, transdermal administration, mucosal administration, rectal administration, vaginal administration, etc.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] The aryl dicarboxylic acid compounds of this invention are novel antibacterial drugs. Experiments have confirmed that these compounds have a strong inhibitory effect on Acinetobacter baumannii. Because this structural type is different from clinically used antibacterial drugs, it is expected to solve the problem of bacterial resistance and has the value for in-depth research and the potential for clinical application. Detailed Implementation

[0022] The present invention will be further described below with reference to specific embodiments. However, the embodiments are only for illustrating the present invention and are not intended to limit the present invention. Unless otherwise specified, the experimental methods used in the embodiments are conventional methods; unless otherwise specified, the materials and reagents used are commercially available.

[0023] I. Preparation and Detection of New Compounds

[0024] Example 1: 5-[3-(3-{3-carboxyl-4-[5-(4-chlorophenyl)thiophene-2-sulfonamido]phenyl}propamido)phenyl]-2-[5-(4-chlorophenyl)thiophene-2-sulfonamido]benzoic acid (In-1)

[0025]

[0026] To a 500 mL round-bottom flask, add 25.0 g (104.8 mmol, 1.2 eq) of 4-chlorophenylboronic acid pinacol ester, 18.34 g (87.3 mmol, 1.0 eq) of 2-iodothiophene, 18.1 g (157.2 mmol, 1.5 eq) of potassium carbonate, 200 mL of 1,4-dioxane, and 50 mL of water, and purge with nitrogen three times. Add tetra(triphenylphosphine)palladium, and purge with nitrogen three times. Heat the reaction mixture under reflux at 130 °C for 4 h, remove the solvent by rotary evaporation, extract with ethyl acetate and water, wash the organic phase with saturated brine, dry to anhydrous sodium sulfate, separate by silica gel column chromatography, and remove the solvent by rotary evaporation to give intermediate C-1 (16.5 g, yield: 97.4%). White solid. ESI-MS (m / z): 195.04 [M+H] + . 1 HNMR (500MHz, Chloroform-d) δ7.56–7.52(m,2H),7.37–7.33(m,2H),7.29(dd,J=3.8,2.4Hz,2H),7.08(dd,J=5.0,3.7Hz,1H).

[0027] Intermediate C-1 (10.0 g, 51.54 mmol, 1.0 eq), ethyl acetate (100 mL), and acetic anhydride (9.68 mL, 103.08 mmol, 2.0 eq) were added to a 250 mL round-bottom flask. Concentrated sulfuric acid (5.49 mL, 103.08 mmol, 2.0 eq) was slowly added dropwise under ice bath conditions. After the addition was complete, the mixture was moved to room temperature and stirred for 24 h. A white solid precipitated upon the addition of saturated brine (150 mL) and ethyl acetate (100 mL). The organic phase was filtered and dried to give intermediate C-2 (14.3 g, yield: 101.3%). White solid. ESI-MS (m / z): 272.99 [MH] - . 1 H NMR (500MHz, Deuterium Oxide) δ7.60 (d, J = 8.7 Hz, 2H), 7.49–7.40 (m, 3H), 7.30 (d, J = 3.9 Hz, 1H).

[0028] Intermediate C-2 (14.3 g, 52.20 mmol, 1.0 eq), thionyl chloride (80 mL, 1.1 mol, 21 eq), and 1 drop of N,N-dimethylformamide were added to a 250 mL round-bottom flask. The reaction mixture was heated under reflux at 85 °C for 8 h, cooled to room temperature, and slowly poured into crushed ice (300 g), producing a large amount of gas. Dichloromethane was then added for extraction, and the organic phase was dried over anhydrous sodium sulfate. The solvent was removed by rotary evaporation to obtain intermediate C-3 (9.3 g, yield: 60.6%), a yellow-green solid. 1 HNMR (500MHz, Chloroform-d) δ7.84 (d, J = 4.1 Hz, 1H), 7.58–7.54 (m, 2H), 7.46–7.42 (m, 2H), 7.29 (d, J = 4.1 Hz, 1H).

[0029] To a 100 mL round-bottom flask, add methyl 2-amino-5-iodobenzoate (2 g, 7.22 mmol, 1.0 eq), dichloromethane (20 mL), and pyridine (1.16 mL, 14.44 mmol, 2.0 eq). Dissolve intermediate C-3 (3.16 g, 10.83 mmol, 1.5 eq) in dichloromethane (20 mL) and add it dropwise to the reaction mixture. Stir the mixture at room temperature for 12 h. Remove the solvent by rotary evaporation, extract with dilute hydrochloric acid (1 M, 20 mL), dry to anhydrous sodium sulfate, separate by silica gel column chromatography, and remove the solvent by rotary evaporation to obtain intermediate C-4 (3.8 g, yield: 98.8%). White solid. ESI-MS (m / z): 531.46 [MH] - . 1HNMR(500MHz,Chloroform-d)δ10.73(s,1H),8.27(d,J=2.2Hz,1H),7.79(dd,J=8.8,2.2Hz,1H),7 .61–7.54(m,2H),7.46(d,J=8.6Hz,2H),7.37(d,J=8.6Hz,2H),7.15(d,J=3.9Hz,1H),3.90(s,3H).

[0030] To a 50 mL round-bottom flask, add intermediate C-4 (1 g, 1.88 mmol, 1.0 eq), acrylic acid (516 μL, 7.52 mmol, 4.0 eq), triethylamine (2.6 mL, 18.8 mmol, 10 eq), and N,N-dimethylformamide (15 mL). Purge the mixture three times with nitrogen. Add 1,1-bis(diphenylphosphine-ferrocene)palladium dichloride (273 mg, 0.376 mmol, 0.2 eq), and purge the mixture three times with nitrogen. Heat the reaction mixture at 100 °C for 2 h. Add dilute hydrochloric acid, extract with ethyl acetate, dry the organic phase to anhydrous sodium sulfate, separate by silica gel column chromatography, and remove the solvent by rotary evaporation to obtain intermediate C-5 (480 mg, yield: 53.5%). White solid. ESI-MS (m / z): 476.21 [MH] - . 1 H NMR (500MHz, DMSO-d6) δ8.07(s,1H),7.99–7.89(m,1H),7.70(t,J=11.1Hz,3H),7.60–7.47(m,5H),6.49(d,J=16.0Hz,1H),3.83(s,3H).

[0031] Intermediate C-5 (450 mg, 0.94 mmol, 1.0 eq) was dissolved in dichloromethane (5 mL) and methanol (5 mL), and dry palladium on carbon (100 mg, palladium content: 10%) was added. The mixture was placed in a hydrogenation reactor (H2: 0.4 MPa) and stirred for 4 h. The mixture was filtered through diatomaceous earth, the filtrate was evaporated to dryness, and the residue was separated by silica gel column chromatography. The solvent was removed by rotary evaporation to obtain intermediate C-6 (400 mg, yield: 88.5%). It was a white solid. ESI-MS (m / z): 478.24 [MH] - . 1H NMR (500MHz, Pyridine-d5) δ8.07(d,J=8.5Hz,1H),7.96(d,J=2.2Hz,1H),7.83(d,J=4.0Hz,1H),7.59(s,1H),7.47( d,J=8.6Hz,2H),7.39–7.33(m,2H),7.28(d,J=4.0Hz,1H),3.73(s,3H),3.05(t,J=7.5Hz,2H),2.81(t,J=7.5Hz,2H).

[0032] To a 50 mL round-bottom flask, add intermediate C-4 (1 g, 1.88 mmol, 1.0 eq), 3-aminophenylboronic acid (386 mg, 2.82 mmol, 1.5 eq), potassium carbonate (779 mg, 5.64 mmol, 3.0 eq), 1,4-dioxane (20 mL), and water (4 mL), and purge with nitrogen three times. Add tetra(triphenylphosphine)palladium (220 mg, 0.19 mmol, 0.1 eq), purge with nitrogen three times, heat the reaction mixture at 60 °C for 2 h, add dilute hydrochloric acid, extract with ethyl acetate, dry the organic phase to anhydrous sodium sulfate, separate by silica gel column chromatography, and remove the solvent by rotary evaporation to obtain intermediate C-7 (530 mg, yield: 56.6%). White solid. ESI-MS (m / z): 499.45 [M+H] + . 1 HNMR (500MHz, DMSO-d6) δ10.54(s,1H),8.01(s,1H),7.84(d,J=8.6Hz,1H),7.70(dd,J=24.7,6.1Hz,3H),7.60(dd,J=21.4,6.3Hz,2H ), 7.50 (d, J = 8.2Hz, 2H), 7.10 (t, J = 7.8Hz, 1H), 6.82 (s, 1H), 6.76 (d, J = 7.6Hz, 1H), 6.58 (d, J = 8.0Hz, 1H), 5.45 (s, 2H), 3.83 (s, 3H).

[0033] To a 50 mL round-bottom flask, add intermediate C-6 (100 mg, 0.21 mmol, 1.0 eq), intermediate C-7 (133 mg, 0.25 mmol, 1.2 eq), N-methylimidazole (60 μL, 0.375 mmol, 3.5 eq), and dichloromethane (5 mL), and stir for 5 minutes. Add N,N,N,N-tetramethylchloromethanesulfonyl hexafluorophosphate (88 mg, 0.315 mmol, 1.5 eq), and stir the reaction mixture at room temperature for 12 hours. Add 10 mL of water, extract with dichloromethane, and remove the solvent by rotary evaporation of the organic phase to obtain intermediate C-8. Tetrahydrofuran (12.5 mL), methanol (500 μL), and saturated lithium hydroxide solution (2 mL) were added. The reaction mixture was stirred at room temperature for 12 h. The pH was adjusted to acidic by adding dilute hydrochloric acid (1 M, 20 mL). Extraction was performed with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and separated by silica gel column chromatography. The solvent was removed by rotary evaporation to give the final product In-1 (103 mg, yield: 52.7%). White solid. ESI-MS (m / z): 930.05 [MH] - . 1 H NMR (700MHz, DMSO-d6) δ10.14(d,J=5.4Hz,1H),8.13(d,J=2.4Hz,1H),7.81(d,J=3 .3Hz,2H),7.68(d,J=8.6Hz,3H),7.65(d,J=8.6Hz,2H),7.61(d,J=8.7Hz,1H),7.58 (d,J=3.9Hz,1H),7.56–7.52(m,2H),7.51–7.49(m,1H),7.49–7.43(m,6H),7.31(t ,J=7.9Hz,1H),7.25(d,J=8.2Hz,1H),2.86(t,J=7.7Hz,2H),2.59(t,J=7.7Hz,2H).

[0034] II. Antibacterial Activity Assay

[0035] Example 1: Determination of the inhibitory activity of the compound against Acinetobacter baumannii ATCC 19606

[0036] 1. Preparation of test bacterial suspension

[0037] Acinetobacter baumannii standard strain ATCC 19606 was inoculated into 20 mL of tryptone soybean broth and incubated at 37°C with shaking at 200 rpm for 18 hours. When the clear medium becomes turbid, it indicates significant bacterial proliferation and vigorous growth. At this point, the bacterial suspension was diluted with fresh tryptone soybean broth to achieve an OD600 value between 0.3 and 0.5, and then further diluted 10 times with fresh tryptone soybean broth.5 The test bacterial solution was prepared by doubling the amount of the solution.

[0038] 2. Determine the minimum inhibitory concentration.

[0039] Take a clean, sterile 96-well cell culture plate. Add 200 μL of the prepared test bacterial solution to each well in the first column, and 100 μL of the test bacterial solution to each well in columns two through twelve. Add 4 μL of the pre-prepared 1.6 mg / mL DMSO solution of the test sample to each well in column one (three replicates per sample). Set up a positive control group (i.e., 4 μL of levofloxacin at the same concentration) and a blank control (i.e., no drug added). Starting from the wells in the first column, use an 8-channel micropipette to sequentially add 100 μL of the sample from the previous column to each well in the next column for a 2-fold serial dilution, setting up 12 different concentrations of the compound solution: 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125, 0.0625, 0.03125, and 0.015625 μg / mL. The 96-well cell culture plates were placed in an incubator at 37°C and cultured for 18 hours. The growth of Acinetobacter baumannii in each well was then observed. For each compound, the concentration of the compound in wells where no bacterial growth was observed was the minimum inhibitory concentration (MIC) of that compound. Levofloxacin was used as a positive control.

[0040] 3. The minimum inhibitory concentration (MIC) of compound In-1 against the Acinetobacter baumannii standard strain ATCC19606 was 0.5 μg / mL, and the MIC of levofloxacin was 0.25 μg / mL. Specific data are shown in Table 1.

[0041] Table 1. Minimum inhibitory concentrations of compounds against Acinetobacter baumannii

[0042]

Claims

1. A compound of formula (1) or a pharmaceutically acceptable salt thereof, wherein: R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 independently selected from hydrogen, halogen.

2. The compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, the halogen is selected from fluorine, chlorine, bromine, iodine.

3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from 5-[3-(3-{3-carboxy-4-[5-(4-chlorophenyl)thiophene-2- sulfonamido]phenyl}propionamido)phenyl]-2-[5-(4-chlorophenyl)thiophene-2-sulfonamido]benzoic acid:

4. A pharmaceutical composition, characterized by, which comprises the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 or 2, a pharmaceutically acceptable carrier.

5. Pharmaceutical composition according to claim 4, characterized in that the pharmaceutical composition is selected from injection, tablet, pill, capsule, suspension, emulsion or ointment, and the administration route is selected from intravenous or intramuscular injection, oral administration, transdermal administration, mucosal administration, rectal administration, vaginal administration.

6. Use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 or 2 in the preparation of an antibacterial drug.

7. Use according to claim 6, characterised in that, the use in the preparation of an antibacterial drug refers to the use in the preparation of a drug for treating or preventing infectious diseases of human or animal caused by pathogenic microorganisms such as bacteria, mycoplasma, chlamydia, rickettsia, spirochete and fungus.

8. Use according to claim 7, characterised in that, the bacteria include Acinetobacter baumannii, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Enterococcus faecium, Clostridium difficile, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, the mycoplasma includes Mycoplasma pneumoniae, Ureaplasma urealyticum, Mycoplasma hominis, Mycoplasma genitalium, the chlamydia includes Chlamydia pneumoniae, Chlamydia psittaci, Chlamydia trachomatis and Chlamydia abortus.

9. Use according to claim 8, characterised in that, the Acinetobacter baumannii includes drug-sensitive Acinetobacter baumannii, multi-drug resistant Acinetobacter baumannii, pan-drug resistant Acinetobacter baumannii, carbapenem-resistant Acinetobacter baumannii, and pandrug-resistant Acinetobacter baumannii, the Staphylococcus aureus includes methicillin-sensitive Staphylococcus aureus and methicillin-resistant Staphylococcus aureus, the Staphylococcus epidermidis includes methicillin-sensitive Staphylococcus epidermidis and methicillin-resistant Staphylococcus epidermidis, the Enterococcus faecalis includes vancomycin-sensitive Enterococcus faecalis and vancomycin-resistant Enterococcus faecalis, the Enterococcus faecium includes vancomycin-sensitive Enterococcus faecium and vancomycin-resistant Enterococcus faecium, and the mycoplasma is selected from drug-sensitive Mycoplasma pneumoniae or drug-resistant Mycoplasma pneumoniae.

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