A bis-aminoguanidine derivative, its preparation and use

Abstract

The application relates to the technical field of drug synthesis, and particularly discloses a diamino guanidine derivative, discloses a structural general formula of the derivative, and provides a preparation method and application of the derivative. The derivative has good inhibiting and killing effects on pseudomonas aeruginosa, the preparation method is simple, the reaction condition is mild, and the derivative has the possibility of being developed into a drug for inhibiting and killing pseudomonas aeruginosa and treating diseases caused by pseudomonas aeruginosa infection.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of drug synthesis, and particularly relates to a diamino guanidine derivative and a preparation method and application thereof. BACKGROUND

[0002] Pseudomonas aeruginosa (PA), commonly known as green pyocyanic bacillus, is widely distributed in the environment and can be detected on the body surface and intestinal tract of normal humans and animals. Pseudomonas aeruginosa is a gram-negative bacillus with a size of (0.5-0.7) μm x (1.5-3.0) μm, and can form single, double or short chain, endogenous single flagellum. It is active under dark field microscopy or phase contrast microscopy, is an aerobic bacterium, does not produce spores, and has an optimal growth temperature of 30-37℃. The characteristics that it does not grow at 4℃ but can grow at 42℃ can be used for identification. It can grow on ordinary culture medium, and most strains can produce water-soluble pigments such as blue pyocyanin and blue-green fluorescein, and a transparent hemolytic ring appears on blood plates. The bacterium contains O antigen, H antigen, sandwich antigen and R antigen. The O antigen can be used for typing.

[0003] Pseudomonas aeruginosa is an opportunistic pathogen with multiple virulence factors, including LPS, quorum sensing, two-component system, six secretion systems, outer membrane vesicles (OMV), CRISPR-Cas, etc., which enable it to have strong adaptation and invasion ability, and it poses a great threat to people and various animals with low immunity or damage, and can widely invade multiple organs and tissues of the body. Pseudomonas aeruginosa can cause wound infection, burn infection, septicemia, pneumonia, etc., and is considered to be the main cause of morbidity and mortality in patients with cystic fibrosis (CF), and is also one of the main causes of nosocomial infection. The threat of Pseudomonas aeruginosa is global: Pseudomonas aeruginosa is the most common cause of hospital-acquired infection in Spain, and Pseudomonas aeruginosa accounts for 10.5% of clinical isolated bacterial infections; Pseudomonas aeruginosa is the fourth most common pathogen in Europe, and almost causes 9% of nosocomial infections; in the United States, 7.1% of healthcare-associated infections are caused by Pseudomonas aeruginosa; data from the National Bacterial Drug Resistance Monitoring Network (CHINET) shows that Pseudomonas aeruginosa is the fourth most common nosocomial infection pathogen in China.

[0004] Pseudomonas aeruginosa has intrinsic resistance to most antibiotics by limiting outer membrane permeability, efflux pumps, producing inactivating enzymes, etc., and has the ability to rapidly acquire drug resistance, making it more challenging to treat Pseudomonas aeruginosa infection. As one of the ESKAPE pathogens (including Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species), the World Health Organization listed Pseudomonas aeruginosa with carbapenem antibiotic resistance as a "key" pathogen in 2017 and a "high concern" pathogen in 2024, and there is an urgent need for new antibacterial drugs in the clinic. Therefore, it is necessary to find new drugs against Pseudomonas aeruginosa. SUMMARY

[0005] The first object of the present application is to provide a diamino guanidine derivative capable of inhibiting and killing Pseudomonas aeruginosa to solve the above problems.

[0006] The second object of the present application is to provide a preparation method of the above derivative.

[0007] The third object of the present application is to provide the use of the above diamino guanidine derivative.

[0008] The object of the present application is realized by the following technical solutions:

[0009] A diamino guanidine derivative, characterized in that the derivative has the following general structure:

[0010]

[0011] In the general formula, R is chlorobenzyl or alkyl.

[0012] As a preferred, in the general formula, R is 2-chlorobenzyl 3-chlorobenzyl 4-chlorobenzyl 2,4-dichlorobenzyl 3,4-dichlorobenzyl or n-heptyl

[0013] As a preferred, the derivative is (2E, 2'E)-2,2'-((5-(2-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carboxamide),

[0014] or (2E, 2'E)-2,2'-((5-(3-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodimide),

[0015] or (2E, 2'E)-2,2'-((5-(4-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodimide),

[0016] or (2E, 2'E)-2,2'-((5-(2,4-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodimide);

[0017] or (2E, 2'E)-2,2'-((5-(3,4-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodimide);

[0018] or (2E, 2'E)-2,2'-((5-(heptyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodimide). The method for preparing the above-mentioned bisaminoguanidine derivative comprises the following steps:

[0019] (1) under basic conditions and in the presence of KI, R-Br is reacted with 5-hydroxyisophthalaldehyde in an organic solvent 1, after cooling, the reaction is quenched by adding water, and the separated solid is obtained; solid;

[0020] (2) under acidic conditions, and excess are added in an organic solvent 2, the reaction is heated, after cooling, the solid is separated or after adding hydrochloric acid, the solid is separated, and the obtained solid is washed and dried to obtain the bisaminoguanidine derivative;

[0021] R is chlorobenzyl or alkyl.

[0022] As preferred, R is 2-chlorobenzyl, 3-chlorobenzyl, 4-chlorobenzyl, 2,4-dichlorobenzyl, 3,4-dichlorobenzyl or n-heptyl.

[0023] As preferred, in step (1), the base used in the basic conditions is potassium carbonate, the molar ratio of R-Br to 5-hydroxyisophthalaldehyde is 1:1, the organic solvent 1 is anhydrous acetonitrile, the reaction is carried out under an oil bath at 80°C, and the reaction time is 40 min-5 h.

[0024] As preferred, in step (2), the acid used in the acidic conditions is hydrochloric acid, the organic solvent 2 is anhydrous ethanol, the reaction is carried out under an oil bath at 80°C, and the reaction time is 1 h.

[0025] The above-mentioned bisaminoguanidine derivative is used in the preparation of a drug for inhibiting or killing Pseudomonas aeruginosa.

[0026] Use of the above-mentioned diamino guanidine derivative in the preparation of a drug for preventing and treating diseases caused by Pseudomonas aeruginosa infection.

[0027] Further, the diseases are wound infection, burn infection, septicemia, pneumonia caused by Pseudomonas aeruginosa in human or animals.

[0028] Compared with the prior art, the present application has the following advantages:

[0029] The diamino guanidine derivative of the present application has good inhibitory and killing effects on Pseudomonas aeruginosa, and has the possibility of being developed into a drug for inhibiting and killing Pseudomonas aeruginosa and a drug for treating diseases caused by Pseudomonas aeruginosa infection, and the preparation method is simple and the reaction conditions are mild. BRIEF DESCRIPTION OF DRAWINGS

[0030] Figure 1 is the time-kill curve of 9L ((2E, 2'E)-2,2'-((5-(2-chlorobenzyloxy)-1,3-phenylene) bis(methylene)) bis(hydrazino-1-carboxamide))。 DETAILED DESCRIPTION

[0031] The preferred embodiments of the present application are described below, and it should be understood that the preferred embodiments described herein are only used to illustrate and explain the present application, and are not used to limit the present application.

[0032] Unless otherwise specified, the raw materials in the embodiments of the present application are purchased through commercial channels, and the specifications of some of the raw materials are as follows: 5-hydroxyisophthalaldehyde: Aladdin, 1g, 97%; 2-chlorobenzyl bromide: Aladdin, 5g, >98.0% (GC); 3-chlorobenzyl bromide: Aladdin, 10g, 97%; 4-chlorobenzyl bromide: Aladdin, 5g, >97.0% (GC); 2,4-dichlorobenzyl bromide: Aladdin, 5g, 98%; 3,4-dichlorobenzyl bromide: Aladdin, 5g, 98%; aminoguanidine hydrochloride: Aladdin, 25g, >98.0% (T).

[0033] The preparation method of the diamino guanidine derivative of the present application mainly includes the following steps:

[0034] (1) under the presence of KI and alkaline conditions, R-Br is reacted with 5-hydroxyisophthalaldehyde in anhydrous acetonitrile at a molar ratio of 1:1, and after cooling, water is added to quench the reaction, and is separated to obtain solid; the alkaline condition uses potassium carbonate, and the heating reaction is carried out at 80℃ oil bath;

[0035] (2) under acid conditions, and excess The reaction is heated in anhydrous ethanol, and solid is precipitated after cooling or after adding hydrochloric acid, and the obtained solid is washed and dried to obtain the diamino guanidine derivative The acid condition uses hydrochloric acid, and the reaction is heated at 80°C in an oil bath.

[0036] In the formula, R is chlorobenzyl or alkyl.

[0037] Specifically, R is 2-chlorobenzyl, 3-chlorobenzyl, 4-chlorobenzyl, 2,4-dichlorobenzyl, 3,4-dichlorobenzyl, or n-heptane.

[0038] Example 1

[0039] Preparation of (2E, 2'E)-2,2'-((5-(2-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodithioate)

[0040] Synthesis of 5-(3-chlorobenzyloxy)isophthalaldehyde: 150 mg (1 mmol) of 5-hydroxyisophthalaldehyde, 553 mg (4 mmol) of potassium carbonate, 17 mg (0.1 mmol) of KI, and 205 mg (1 mmol) of 2-chlorobenzyl bromide were sequentially added to 5 mL of anhydrous acetonitrile, and the reaction solution was stirred and refluxed at 80°C in an oil bath for 40 min. After cooling to room temperature, 60 mL of water was added to quench the reaction, and a large amount of white precipitate was generated. After suction filtration and drying, 5-(3-chlorobenzyloxy)isophthalaldehyde was obtained as a white solid in a yield of 89%.

[0041] Synthesis of (2E, 2'E)-2,2'-((5-(2-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodithioate): 245 mg of 5-(2-chlorobenzyloxy)isophthalaldehyde and 332 mg of aminoguanidine hydrochloride (3 mmol) were suspended in anhydrous ethanol, and the aminoguanidine hydrochloride was in excess. Fifteen drops of 37 wt% concentrated hydrochloric acid were added dropwise, and the reaction was stirred and refluxed at 80°C in an oil bath for 1 h. After cooling to room temperature, a large amount of white solid was precipitated, and the solid was separated. After being washed with cold ethanol twice and then with water twice, the white solid was dried to obtain (2E, 2'E)-2,2'-((5-(2-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodithioate) in a yield of 83%.

[0042] Example 2

[0043] Preparation of (2E, 2'E)-2,2'-((5-(3-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodithioate)

[0044] Synthesis of 5-(3-chlorobenzyloxy)isophthalaldehyde: 150 mg (1 mmol) of 5-hydroxyisophthalaldehyde, 553 mg (4 mmol) of potassium carbonate, 17 mg (0.1 mmol) of KI, and 205 mg (1 mmol) of 2-chlorobenzyl bromide were sequentially added to 5 mL of anhydrous acetonitrile, and the reaction solution was stirred and refluxed at 80°C in an oil bath for 40 min. After cooling to room temperature, 60 mL of water was added to quench the reaction, and a large amount of white precipitate was generated. After suction filtration and drying, 5-(3-chlorobenzyloxy)isophthalaldehyde was obtained as a white solid in a yield of 89%.

[0045] Synthesis of (2E,2'E)-2,2'-((5-(3-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine): Basically the same as Example 1, except that 5-(2-chlorobenzyloxy)isophthalaldehyde was replaced with 5-(3-chlorobenzyloxy)isophthalaldehyde in an amount of 215 mg to give a white solid with a yield of 96%.

[0046] Example 3

[0047] Preparation of (2E,2'E)-2,2'-((5-(4-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine)

[0048] Synthesis of 5-(4-chlorobenzyloxy)isophthalaldehyde: basically the same as Example 1, except that 2-chlorobenzyl bromide was replaced by 4-chlorobenzyl bromide.

[0049] Synthesis of (2E,2'E)-2,2'-((5-(4-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine): Basically the same as Example 1, except that 5-(2-chlorobenzyloxy)isophthalaldehyde was replaced with 5-(4-chlorobenzyloxy)isophthalaldehyde in an amount of 250 mg to give a white solid with a yield of 93.92%.

[0050] Example 4

[0051] Preparation of (2E,2'E)-2,2'-((5-(2,4-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine)

[0052] Synthesis of 5-(2,4-chlorobenzyloxy)isophthalaldehyde: basically the same as Example 1, except that 2-chlorobenzyl bromide was replaced by 2,4-chlorobenzyl bromide in an amount of 240 mg (1 mmol).

[0053] Synthesis of (2E,2'E)-2,2'-((5-(2,4-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine): Basically the same as Example 1, except that 5-(2-chlorobenzyloxy)isophthalaldehyde was replaced with 5-(2,4-chlorobenzyloxy)isophthalaldehyde in an amount of 277 mg to give a white solid with a yield of 94.5%.

[0054] Example 5

[0055] Preparation of (2E,2'E)-2,2'-((5-(3,4-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine)

[0056] Synthesis of 5-(3,4-dichlorobenzyloxy)isophthalaldehyde: substantially the same as Example 1, except that 2-chlorobenzyl bromide was replaced by 3,4-dichlorobenzyl bromide, which was used in an amount of 240 mg (1 mmol).

[0057] Synthesis of (2E,2'E)-2,2'-((5-(3,4-dichlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodithioate): substantially the same as Example 1, except that 5-(2-chlorobenzyloxy)isophthalaldehyde was replaced by 5-(3,4-dichlorobenzyloxy)isophthalaldehyde, which was used in an amount of 298 mg, to obtain a light purple solid with a yield of 51.3%.

[0058] Example 6

[0059] Synthesis of (2E,2'E)-2,2'-((5-(heptyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodithioate)

[0060] Synthesis of 5-(heptyloxy)isophthalaldehyde: 150 mg (1 mmol) of 5-hydroxyisophthalaldehyde, 553 mg (4 mmol) of potassium carbonate, 17 mg (0.1 mmol) of KI and 90 mg (1 mmol) of n-heptyl bromide were sequentially added to 5 mL of anhydrous acetonitrile, and the reaction solution was stirred in an 80°C oil bath for reflux reaction for 5 h, and then cooled to room temperature. After 60 mL of water was added, a large amount of white precipitate was generated, which was suction filtered and air dried to obtain white solid 5-(heptyloxy)isophthalaldehyde with a yield of 87%.

[0061] Synthesis of (2E,2'E)-2,2'-((5-(heptyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodithioate): 227 mg of the obtained 5-(heptyloxy)isophthalaldehyde and 332 mg of aminoguanidine hydrochloride were suspended in anhydrous ethanol, with the aminoguanidine hydrochloride being in excess. Fifteen drops of 37 wt% concentrated hydrochloric acid were added dropwise, and the reaction was stirred in an 80°C oil bath for reflux reaction for 1 h. After cooling to room temperature, 1 M hydrochloric acid solution was added dropwise, and white flocculent precipitate was generated. The solid was separated, washed with petroleum ether for 2 times, and oven dried to obtain yellowish solid (2E,2'E)-2,2'-((5-(heptyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodithioate) with a yield of 77.1%.

[0062] Using the above synthesis method, a series of bisaminoguanidine derivatives were prepared. The structural formula, molecular formula and chemical name of each compound are shown in Table 1, and the corresponding nuclear magnetic resonance hydrogen spectrum and nuclear magnetic resonance carbon spectrum and high resolution mass spectrum data are shown in Table 2.

[0063] Table 1 Structure, molecular formula and corresponding chemical name of bisaminoguanidine derivatives

[0064]

[0065]

[0066] Table 2 Nuclear magnetic resonance hydrogen spectrum, nuclear magnetic resonance carbon spectrum and high resolution mass spectrum data of the diamino guanidine derivative

[0067]

[0068]

[0069] Example 7

[0070] Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) test of the diamino guanidine derivative

[0071] The minimum inhibitory concentration and minimum bactericidal concentration of Pseudomonas aeruginosa (ATCC 27853, ATCC 9027, CMCC (B) 10104, clinical strain 4531) were determined by micro-broth dilution method. Compound 9L, 9M, 9N, 9O, 9P and 9X were prepared into 1280 μg / mL DMSO solution, 10 μL of the compound solution was mixed with 190 μL of MH broth in a 96-well plate, 100 μL of the mixture was serially diluted by two times in a 96-well plate, 100 μL of 1000-fold diluted bacterial solution was inoculated, and after incubation at 37°C for 16-20 hours, the clarity of the mixture was observed by naked eye, and the lowest concentration with clear mixture and no bacterial precipitation at the bottom of the well was the minimum inhibitory concentration of the compound. 100 μL of the bacterial solution without bacterial growth was uniformly coated on the MH agar medium, and after incubation at 37°C for 24 hours, the concentration without bacterial growth was the minimum bactericidal concentration of the compound. The MH broth without the addition of the compound was used as a blank control group, and ceftazidime and levofloxacin were used as positive control drugs. The minimum inhibitory concentration and minimum bactericidal concentration (MIC / MBC) results are shown in Table 3.

[0072] Table 3 Minimum inhibitory concentration / minimum bactericidal concentration (MIC / MBC) of the diamino guanidine derivative on four strains of Pseudomonas aeruginosa

[0073]

[0074] As can be seen from Table 3, each diamino guanidine derivative prepared in the present application has a strong inhibitory effect on Pseudomonas aeruginosa; the minimum bactericidal concentration (MBC) of most of the compounds is in the range of 1-4 times the minimum inhibitory concentration (MIC), showing bactericidal effect; among them, 9L performs the best.

[0075] Example 8

[0076] Bactericidal kinetics test of 9L

[0077] The time-kill curves of different concentrations of 9L against Pseudomonas aeruginosa ATCC 27853 were detected. After ATCC 27853 was cultured in MH broth to the logarithmic growth phase, it was diluted into a bacterial suspension of about 1 x 10 6 CFU / mL. Compound 9L (4, 8, 16 μg / mL, i.e. 2 MIC, 4 MIC, 8 MIC, respectively) and levofloxacin (1 μg / mL, i.e. 1 MBC) were added to the bacterial suspension, respectively, and an equal amount of DMSO was used as a blank control, and they were cultured at 37°C with shaking. At the specified time intervals (0, 2, 4, 8, 12, 24 h), samples were taken, respectively, and counted using a 10-fold dilution plate method. The time-kill curves were plotted with the logarithm of the number of bacteria per milliliter as the ordinate and the culture time as the abscissa as shown in Figure 1 .

[0078] As can be seen from Figure 1 , the number of bacteria at 24 h was greater than the inoculum and less than the blank control group for 1 μg / mL of levofloxacin, which had a bacteriostatic effect. The bactericidal effect of 9L on ATCC 27853 was concentration-dependent, and different concentrations of 9L (4, 8, 16 μg / mL, i.e. 2 MIC, 4 MIC, 8 MIC) had a significant inhibitory or killing effect on ATCC 27853, of which 2 MIC and 4 MIC had an inhibitory effect with the number of bacteria at 24 h being greater than the inoculum and less than the blank control group; 8 MIC had a significant bactericidal effect with the number of bacteria at 24 h being less than the inoculum.

[0079] The above only describes preferred embodiments of the present application and is not intended to limit the present application. Although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art will still be able to modify the technical solutions described in the foregoing embodiments or make equivalent replacements to some technical features. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A bis-amino guanidine derivative, characterized by, The derivative has the following general structure: In the general formula, R is 2-chlorobenzyl, 3-chlorobenzyl, 4-chlorobenzyl, 2,4-dichlorobenzyl, 3,4-dichlorobenzyl or n-heptyl.

2. The bis-aminoguanidine derivative according to claim 1, characterized in that, The derivative is (2E, 2'E)-2,2'-((5-(2-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodithioate), or (2E, 2'E)-2,2'-((5-(3-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodithioate), or (2E, 2'E)-2,2'-((5-(4-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodithioate), or (2E, 2'E)-2,2'-((5-(2,4-dichlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodithioate); or (2E, 2'E)-2,2'-((5-(3,4-dichlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodithioate); or (2E, 2'E)-2,2'-((5-(heptyloxy)-1,3-phenylene)bis(methylene))bis(hydrazinyl-1-carbodithioate).

3. The method of producing the bisaminoguanidine derivative according to claim 1, characterized by, The method comprises the following steps: (1) R-Br and 5-hydroxyisophthalaldehyde were heated in an organic solvent in the presence of KI under basic conditions, and after cooling, water was added to quench the reaction, and the product was separated solid; (2) under acidic conditions, with excess The reaction is heated in organic solvent II, and solid is precipitated after cooling or after adding hydrochloric acid. The obtained solid is washed and dried to obtain the diamino guanidine derivative. R is 2-chlorobenzyl, 3-chlorobenzyl, 4-chlorobenzyl, 2,4-dichlorobenzyl, 3,4-dichlorobenzyl or n-heptyl.

4. The method of claim 3, wherein the method is characterized by, In step (1), the base used in the basic condition is potassium carbonate, the molar ratio of R-Br to 5-hydroxyisophthalaldehyde is 1:1, the organic solvent I is anhydrous acetonitrile, and the reaction is carried out in an 80°C oil bath, and the reaction time is 40 min-5 h.

5. The method for preparing the bisaminoguanidine derivatives according to claim 4, characterized in that: In step (2), the acid used in the acidic condition is hydrochloric acid, the organic solvent II is anhydrous ethanol, and the reaction is carried out in an 80°C oil bath, and the reaction time is 1 h.

6. The use of the bisaminoguanidine derivative of claim 1 in the preparation of a drug for inhibiting or killing Pseudomonas aeruginosa.

7. The use of the bisaminoguanidine derivative of claim 1 in the preparation of a drug for preventing and treating diseases caused by Pseudomonas aeruginosa infection.

8. Use according to claim 7, characterized in that, The diseases are wound infection, burn infection, septicemia, pneumonia caused by Pseudomonas aeruginosa in humans or animals.

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