Diaminoguanidine derivative as well as preparation method and application thereof

Abstract

The invention relates to the technical field of drug synthesis, and particularly discloses a diaminoguanidine derivative and a structural general formula of the derivative. The invention also provides a preparation method and application thereof. The derivative has a good inhibiting and killing effect on pseudomonas aeruginosa, the preparation method is simple, reaction conditions are mild, and the derivative has the possibility of being developed into drugs for inhibiting and killing pseudomonas aeruginosa and drugs for treating diseases caused by pseudomonas aeruginosa infection.

Description

Technical Field

[0001] The invention belongs to the technical field of drug synthesis, and specifically relates to a diaminoguanidine derivative and a preparation method and application thereof. Background Art

[0002] Pseudomonas aeruginosa (PA), commonly known as Pseudomonas aeruginosa, belongs to the Pseudomonas family and is widely distributed in the environment. It can also be detected on the body surface and intestines of normal humans and animals. Pseudomonas aeruginosa is a Gram-negative bacillus with a body size of (0.5-0.7) μm × (1.5-3.0) μm. It is single, double or short chain, with a single flagellum at the end. It can be observed under a dark field microscope or phase contrast microscope that the bacteria are active in movement. It is an aerobic bacterium and does not produce spores. The optimal growth temperature is 30-37°C. It does not grow at 4°C but can grow at 42°C. This feature can be used for identification. It can grow on ordinary culture media. Most strains can produce water-soluble pigments, such as blue pyocyanin and blue-green fluorescein, and transparent hemolytic rings will appear on blood plates. The bacteria contain O antigen, H antigen, sandwich antigen, R antigen, etc. 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 adaptability and invasion ability. It poses a great threat to people with low immunity or damage and a variety of animals, and can widely invade multiple organs and tissues of the body. Pseudomonas aeruginosa can cause trauma infection, burn infection, sepsis, 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 infections in Spain, accounting for 10.5% of clinically isolated bacterial infections; Pseudomonas aeruginosa is the fourth most common pathogen in European hospitals, causing almost 9% of nosocomial infections; in the United States, 7.1% of healthcare-associated infections are caused by Pseudomonas aeruginosa; data from the National Bacterial Resistance Monitoring Network (CHINET) show that Pseudomonas aeruginosa is the fourth most common pathogen of nosocomial infections in China.

[0004] Pseudomonas aeruginosa has intrinsic resistance to most antibiotics by limiting outer membrane permeability, efflux pumps, producing inactivating enzymes, etc., and it has the ability to quickly acquire resistance, making the treatment of Pseudomonas aeruginosa infection more challenging. Pseudomonas aeruginosa is 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 resistance as a "critical" pathogen in 2017 and a "high concern" pathogen in 2024. New antibacterial drugs are urgently needed in clinical practice. Therefore, it is necessary to find new anti-Pseudomonas aeruginosa drugs. Summary of the invention

[0005] The first purpose of the present invention is to solve the above problems and provide a biaminoguanidine derivative capable of inhibiting and killing Pseudomonas aeruginosa.

[0006] The second object of the present invention is to provide a method for preparing the above derivatives.

[0007] The third object of the present invention is to provide the use of the above-mentioned bisaminoguanidine derivatives.

[0008] The purpose of the present invention is specifically achieved through the following technical solutions:

[0009] A bisaminoguanidine derivative, characterized in that the derivative has the following general structural formula:

[0010]

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

[0012] Preferably, in the general formula, R is 2-chlorobenzyl 3-Chlorobenzyl 4-Chlorobenzyl 2,4-Dichlorobenzyl 3,4-Dichlorobenzyl or n-heptyl

[0013] Preferably, the derivative is (2E,2'E)-2,2'-((5-(2-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine),

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

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

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

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

[0018] or (2E,2'E)-2,2'-((5-(heptyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine). The preparation method of the above-mentioned bisaminoguanidine derivatives comprises the following steps:

[0019] (1) Under alkaline conditions and in the presence of KI, R-Br reacts with 5-hydroxyisophthalaldehyde in an organic solvent, and after cooling, water is added to quench the reaction, and the obtained solid;

[0020] (2) Under acidic conditions, With overdose The reaction is heated in an organic solvent II, and a solid is precipitated after cooling or after adding hydrochloric acid, and the obtained solid is washed and dried to obtain a bisaminoguanidine derivative;

[0021] R is chlorobenzyl or alkyl.

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

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

[0024] Preferably, in step (2), the acid used in the acidic condition is hydrochloric acid, the second organic solvent is anhydrous ethanol, and the reaction is carried out in an oil bath at 80° C. for 1 h.

[0025] Application of the above-mentioned bisaminoguanidine derivatives in the preparation of drugs for inhibiting or killing Pseudomonas aeruginosa.

[0026] The application of the above-mentioned bisaminoguanidine derivatives in the preparation of drugs for preventing and treating diseases caused by Pseudomonas aeruginosa infection.

[0027] Furthermore, the disease is wound infection, burn infection, sepsis, or pneumonia caused by Pseudomonas aeruginosa in humans or animals.

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

[0029] The bisaminoguanidine derivatives of the invention have good inhibitory and killing effects on Pseudomonas aeruginosa, and the preparation method thereof is simple and the reaction conditions are mild. The derivatives have the possibility of being developed into drugs for inhibiting and killing Pseudomonas aeruginosa and drugs for treating diseases caused by Pseudomonas aeruginosa infection. BRIEF DESCRIPTION OF THE DRAWINGS

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

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

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

[0033] The preparation method of the bisaminoguanidine derivatives of the present invention mainly comprises the following steps:

[0034] (1) Under alkaline conditions and in the presence of KI, R-Br and 5-hydroxyisophthalaldehyde are heated in anhydrous acetonitrile at a molar ratio of 1:1, and after cooling, water is added to quench the reaction, and the obtained Solid; potassium carbonate was used as alkaline condition, and the heating reaction was carried out in an oil bath at 80°C;

[0035] (2) Under acidic conditions, With overdose Heat the reaction in anhydrous ethanol, precipitate solid after cooling or add hydrochloric acid to precipitate solid, wash and dry the obtained solid to obtain the bisaminoguanidine derivative The acidic condition was hydrochloric acid, and the heating reaction was carried out in an oil bath at 80°C;

[0036] Wherein R is chlorobenzyl or alkyl.

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

[0038] Example 1

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

[0040] Synthesis of 5-(2-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 added to 5 mL of anhydrous acetonitrile in sequence. The reaction solution was stirred and refluxed in an 80°C oil bath for 40 min, then cooled to room temperature, and 60 ml of water was added to quench the reaction, generating a large amount of white precipitate, which was filtered and dried to obtain white solid 5-(2-chlorobenzyloxy)isophthalaldehyde with a yield of 89%.

[0041] Synthesis of (2E,2'E)-2,2'-((5-(2-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine): 245 mg of 5-(2-chlorobenzyloxy)isophthalaldehyde and 332 mg of aminoguanidine hydrochloride (3 mmol) were suspended in anhydrous ethanol, wherein the aminoguanidine hydrochloride was in excess, 15 drops of 37wt% concentrated hydrochloric acid were added dropwise, and the mixture was stirred and refluxed in an 80°C oil bath for 1 hour. The mixture was cooled to room temperature, and a large amount of white solid was precipitated. The solid was separated, washed twice with cold ethanol, and then washed twice with water, and dried to obtain a white solid, namely (2E,2'E)-2,2'-((5-(2-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine), with a yield of 83%.

[0042] Example 2

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

[0044] Synthesis of 5-(3-chlorobenzyloxy)isophthalaldehyde: Basically the same as Example 1, except that 2-chlorobenzyl bromide is replaced by 3-chlorobenzyl bromide.

[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 obtain 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 is 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 obtain 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 is 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 obtain 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-chlorobenzyloxy)isophthalaldehyde: The method is basically the same as that in Example 1, except that 2-chlorobenzyl bromide is replaced by 3,4-chlorobenzyl bromide in an amount of 240 mg (1 mmol).

[0057] Synthesis of (2E,2'E)-2,2'-((5-(3,4-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-(3,4-chlorobenzyloxy)isophthalaldehyde in an amount of 298 mg to obtain a lavender solid with a yield of 51.3%.

[0058] Example 6

[0059] Preparation of (2E,2'E)-2,2'-((5-(heptyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine)

[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-heptane bromide were added to 5 mL of anhydrous acetonitrile in sequence. The reaction solution was stirred and refluxed in an oil bath at 80°C for 5 h, then cooled to room temperature and 60 ml of water was added to generate a large amount of white precipitate. The precipitate was filtered and dried to obtain 5-(heptyloxy)isophthalaldehyde as a white solid with a yield of 87%.

[0061] Synthesis of (2E,2'E)-2,2'-((5-(heptyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine): 227 mg of the obtained 5-(heptyloxy)isophthalaldehyde and 332 mg of aminoguanidine hydrochloride were suspended in anhydrous ethanol, wherein the aminoguanidine hydrochloride was in excess, 15 drops of 37wt% concentrated hydrochloric acid were added dropwise, and the reaction was stirred and refluxed in an 80°C oil bath for 1 hour. The reaction was cooled to room temperature, and 1M hydrochloric acid solution was added dropwise to generate a white flocculent precipitate. The solid was separated, washed with petroleum ether twice, and dried to obtain a slightly yellow solid, namely (2E,2'E)-2,2'-((5-(heptyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine), with a yield of 77.1%.

[0062] A series of bisaminoguanidine derivatives were prepared using the above synthesis method. The structural formula, molecular formula and chemical name of each compound are shown in Table 1, and the corresponding H NMR spectrum, C NMR spectrum and high-resolution mass spectrum data are shown in Table 2.

[0063] Table 1 Structures, molecular formulas and corresponding chemical names of bisaminoguanidine derivatives

[0064]

[0065]

[0066] Table 2 H NMR, C NMR and high resolution mass spectrometry data of bisaminoguanidine derivatives

[0067]

[0068]

[0069] Example 7

[0070] Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) test of bisaminoguanidine derivatives

[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 the microbroth dilution method. Compounds 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 MH broth in a 96-well plate. 100 μL of the mixture was diluted two-fold in a 96-well plate, and 100 μL of the 1000-fold diluted bacterial solution was inoculated. After constant temperature incubation at 37°C for 16-20 hours, the clarity of the mixture was observed with the naked eye. The lowest concentration that was clear and had 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 evenly spread on MH agar medium. After incubation at 37°C for 24 hours, the concentration at which no bacterial growth was observed was the minimum bactericidal concentration of the compound. MH broth without adding the compound was used as a blank control group, and ceftazidime and levofloxacin were used as positive control drugs. The results of minimum inhibitory concentration and minimum bactericidal concentration (MIC / MBC) are shown in Table 3.

[0072] Table 3 Minimum inhibitory concentration / minimum bactericidal concentration (MIC / MBC) of bisaminoguanidine derivatives against four strains of Pseudomonas aeruginosa

[0073]

[0074] As can be seen from Table 3, the bisaminoguanidine derivatives prepared in the present invention have a strong inhibitory effect on Pseudomonas aeruginosa; the minimum bactericidal concentration (MBC) of most compounds is within the range of 1-4 times the minimum inhibitory concentration (MIC), showing a bactericidal effect; among them, 9L performed the best.

[0075] Example 8

[0076] Bactericidal kinetics test of 9L

[0077] The time-killing curve of different concentrations of 9L against Pseudomonas aeruginosa ATCC 27853 was drawn. ATCC27853 was cultured in MH broth medium to the logarithmic growth phase and then diluted to about 1×10 6 CFU / mL bacterial suspension. Compound 9L (final concentrations of 4, 8, 16 μg / mL, i.e. 2MIC, 4MIC, 8MIC) and levofloxacin (final concentration of 1 μg / Ml, i.e. 1MBC)) were added to the bacterial suspension, and an equal amount of DMSO was used as a blank control, and cultured at 37°C with shaking. After the specified time intervals (0, 2, 4, 8, 12, 24h), samples were taken and counted using the 10-fold dilution plate method. The time-killing curve was drawn with the logarithm of the number of bacteria per milliliter as the ordinate and the culture time as the abscissa. Figure 1 .

[0078] Depend on Figure 1 It can be seen that the number of bacteria at 1 μg / mL of levofloxacin at 24 hours was greater than the inoculation amount and less than the blank control group, which had an antibacterial effect. The bactericidal effect of 9L on ATCC 27853 was concentration-dependent. 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. Among them, the number of bacteria at 2×MIC and 4×MIC at 24 hours was greater than the inoculation amount and less than the blank control group, which had an inhibitory effect; 8×MIC was less than the inoculation amount at 24 hours, which had a significant bactericidal effect.

[0079] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention is described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A bisaminoguanidine derivative, characterized in that: The derivative has the following general structural formula: In the general formula, R is a chlorobenzyl group or an alkyl group.

2. The bisaminoguanidine derivative according to claim 1, characterized in that: In the general formula, R is 2-chlorobenzyl, 3-chlorobenzyl, 4-chlorobenzyl, 2,4-dichlorobenzyl, 3,4-dichlorobenzyl or n-heptyl.

3. The bisaminoguanidine derivative according to claim 2, characterized in that: The derivative is (2E,2'E)-2,2'-((5-(2-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine), or (2E,2'E)-2,2'-((5-(3-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine), or (2E,2'E)-2,2'-((5-(4-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine), or (2E,2'E)-2,2'-((5-(2,4-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine); or (2E,2'E)-2,2'-((5-(3,4-chlorobenzyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine); or (2E,2'E)-2,2'-((5-(heptyloxy)-1,3-phenylene)bis(methylene))bis(hydrazino-1-amidine).

4. The method for preparing the bisaminoguanidine derivatives according to claim 1, characterized in that: The method comprises the following steps: (1) Under alkaline conditions and in the presence of KI, R-Br reacts with 5-hydroxyisophthalaldehyde in an organic solvent, and after cooling, water is added to quench the reaction, and the obtained solid; (2) Under acidic conditions, With overdose The reaction is heated in an organic solvent II, and a solid is precipitated after cooling or after adding hydrochloric acid, and the obtained solid is washed and dried to obtain a bisaminoguanidine derivative; R is chlorobenzyl or alkyl.

5. The method for preparing the bisaminoguanidine derivatives according to claim 4, characterized in that: R is 2-chlorobenzyl, 3-chlorobenzyl, 4-chlorobenzyl, 2,4-dichlorobenzyl, 3,4-dichlorobenzyl or n-heptyl.

6. The method for preparing the bisaminoguanidine derivatives according to claim 5, characterized in that: In step (1), the base used in the alkaline condition is potassium carbonate, the molar ratio of R-Br to 5-hydroxyisophthalaldehyde is 1:1, the first organic solvent is anhydrous acetonitrile, the reaction is carried out in an oil bath at 80°C, and the reaction time is 40min-5h.

7. The method for preparing the bisaminoguanidine derivatives according to claim 6, characterized in that: In step (2), the acid used in the acidic condition is hydrochloric acid, the second organic solvent is anhydrous ethanol, and the reaction is carried out in an oil bath at 80° C. for 1 h.

8. Use of the bisaminoguanidine derivatives according to claim 1 in the preparation of drugs for inhibiting or killing Pseudomonas aeruginosa.

9. Use of the bisaminoguanidine derivatives according to claim 1 in the preparation of drugs for preventing and treating diseases caused by Pseudomonas aeruginosa infection.

10. The use according to claim 9, characterized in that: The disease is wound infection, burn infection, sepsis, pneumonia caused by Pseudomonas aeruginosa in humans or animals.

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