CN111773216A

CN111773216A - Application of C-JUN N-terminal kinase inhibitor SU3327

Info

Publication number: CN111773216A
Application number: CN202010744897.4A
Authority: CN
Inventors: 陈洪亮
Original assignee: Individual
Current assignee: Individual
Priority date: 2020-07-29
Filing date: 2020-07-29
Publication date: 2020-10-16
Also published as: CN113491692B; CN113491692A

Abstract

The invention relates to application of a C-JUN N-terminal kinase (JNK) inhibitor SU3327, wherein the C-JUN N-terminal kinase inhibitor SU3327 is used for bacteriostasis of different animal and human species. The C-JUN N-terminal kinase inhibitor SU3327 is used for killing and inhibiting bacteria of a strain obtained by separating human and/or animal bodies or clinically separating the strain from infected persons and then culturing the strain.

Description

Application of C-JUN N-terminal kinase inhibitor SU3327

Technical Field

The invention relates to application of a C-JUN N-terminal kinase (JNK) inhibitor SU 3327.

Background

Antibiotics are the most effective drugs found in humans from soil and metabolites of various microorganisms to treat bacterial infections. With the abuse of antibiotics, bacteria begin to have drug resistance to the antibiotics and become drug-resistant bacteria, thereby improving the difficulty of treating diseases of human and animals. China releases a feed 'banning' order (No. 194 bulletin) in 2019, definitely prohibits the use of feed additives containing growth-promoting drugs in the breeding industry, actively promotes the development direction of realizing feed 'banning', breeding 'resistance reduction' and product 'no resistance' in the breeding industry, and encourages the research and development of safe and efficient novel research and development of antibacterial drugs to replace traditional antibiotics. The development of antibiotics requires screening in soil and various products of microorganisms therein, which requires high cost and a very long time, and the growth rate of bacterial resistance is much faster than the development rate of new antibiotics.

SU3327, a c-Jun N-terminal kinase (JNK) inhibitor, is useful for the treatment of diabetes. The molecular formula is C5H3N5O2S3, and the structural formula of the molecular weight 261.313 is shown in figure 1.

SU3327 is disclosed as a reference for preclinical study drugs for the treatment of diabetes as follows:

j Med Chem journal (2009 Apr 9), titled Design, Synthesis, and Structure-Activity Relationship of Substrate compatibility, Selective, and in Vivo Activity triazole and clinical inhibitors of the c-Jun N-Terminal Kinase, the main contents of which are: SU3327 is named as compound 9 in the article and is an artificially synthesized JNK inhibitor. Protein kinases account for 2% of the mammalian genome and catalyze the transfer of gamma-phosphoryl of ATP to specific protein substrates. The c-Jun N-terminal kinase (JNK) is a series of serine/threonine protein kinases of the mitogen-activated protein kinase (MAPK) family. Upregulation of JNK activity has been associated with a number of diseases, such as type-2 diabetes, obesity, cancer, and stroke. JNK inhibitors can inhibit JNK activity, and thus, JNK inhibitors have effects against various diseases.

Acta Pharmacol Sin journal (2014 Mar) titled TNF- α indeces CXCL1 chemokinexpression and release in human vascular endothelial cells in vitro via twomodicidin signaling pathwalls, the main contents of which are: the article found that TNF- α (2, 5 ng/mL) induced the release of CXCL1 and mRNA expression in cells in a concentration and time dependent manner. TNF- α causes JNK, p38 MAPK, PI3K and Akt activation, whereas pretreatment with JNK inhibitors (SP 600125 and SU 3327), p38 MAPK inhibitors (SB 202190) or PI-3K inhibitors (LY 294002) significantly inhibited TNF- α -induced release of CXCL1 from cells, in which SU3327 was used as a JNK inhibitor.

Microvasc Res journal (2012 Sep) titled Post-translational adjustment of plant growth factor mRNA by hydrogen peroxide, the main contents of which are: oxidative stressors such as hydrogen peroxide activate multiple kinase pathways, which affect transcription, RNA stability and translation processes. The kinase pathway contributes to the increase of peroxide-induced PLGF mRNA levels, and SU3327, as a JNK inhibitor, can significantly inhibit the increase of peroxide-induced PLGF mRNA levels.

Redox Biol journal (2015 Dec) titled Critical roll of c-jun N-tertiary protein kinase in promoting mitotic dysfunction and ace lever in j oury, the main contents of which are: the article mainly studies the key role of c-jun N-terminal protein kinase (JNK) in promoting mitochondrial dysfunction and acute liver injury. The article uses SU3327 treated mice as a control, i.e. JNK activity is inhibited, whereby it was found that the levels of p-JNK, mitochondrial phosphoprotein and liver damage in CCl4 exposed mice were significantly reduced under SU3327 pretreatment conditions.

Disclosure of Invention

The invention provides application of a C-JUN N-terminal kinase (JNK) inhibitor SU3327, for example, the C-JUN N-terminal kinase inhibitor SU3327 is used for sterilization and bacteriostasis of different animal and human species.

Further: the C-JUN N-terminal kinase inhibitor SU3327 is used for killing and inhibiting bacteria of a strain obtained by separating human and/or animal bodies or clinically separating the strain from infected persons and then culturing the strain.

In some embodiments, the SU3327 has bactericidal activity against a plurality of bacteria including escherichia coli ATCC25922, staphylococcus aureus cmcc (b) 26003, staphylococcus aureus MRSA ATCC43300, acinetobacter baumannii ATCC19606, clostridium sporogenes cmcc (b) 64941, bacillus subtilis cmcc (b) 63501, candida albicans ATCC 10231. Moreover, compared with the aminoglycoside broad-spectrum antibiotic of gentamicin sulfate, SU3327 has better bacteriostatic effect.

Escherichia coli ATCC25922, Staphylococcus aureus CMCC (B) 26003, Staphylococcus aureus MRSAATCC43300, Acinetobacter baumannii ATCC19606, Clostridium sporogenes CMCC (B) 64941, Bacillus subtilis CMCC (B) 63501 and Candida albicans ATCC10231 are all derived from strains isolated from human or/and animal bodies or cultured after being isolated from infected persons clinically.

Drawings

FIG. 1 is a structural formula of an N-terminal kinase inhibitor SU3327 according to the present invention.

Detailed Description

The invention is based on deep learning technology, and the model autonomously finds out potential novel antibiotics from the existing safe medicine database or the safe medicine database in the clinical test stage. The activity of SU3327 with broad-spectrum antibiotics is confirmed by bacterial drug sensitivity experiments. The invention provides a novel antibiotic, which further brings important influence to the field of medicine.

The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of SU3327 against Escherichia coli ATCC25922, Staphylococcus aureus CMCC (B) 26003, Staphylococcus aureus MRSA ATCC43300, Acinetobacter baumannii ATCC19606, Clostridium sporogenes CMCC (B) 64941, Bacillus subtilis CMCC (B) 63501, Candida albicans ATCC10231 were determined using bacterial susceptibility experiments. According to experimental results, SU3327 has good bacteriostatic ability, can be used as a broad-spectrum antibiotic, and effectively reduces the fatality rate of patients infected by pathogenic bacteria.

The following description will mainly describe the drug sensitivity test of SU3327 to various bacteria, i.e., the determination of the Minimum Inhibitory Concentration (MIC) and the Minimum Bactericidal Concentration (MBC), for explaining the present invention. It should be noted that the specific test protocol described below is for illustrative purposes only and does not specifically limit the scope of the present invention.

In the following examples, the protocols used are conventional unless otherwise specified.

In the following examples, materials, reagents and the like used were obtained from commercial sources unless otherwise specified.

1, culture of strain and adjustment of concentration

Selecting pure bacteria solution cultured for 18-24h, dissolving in 2-5ml sterile physiological saline, and adjusting turbidity to 0.5 McLeeb turbidimetric tube.

2, determination of Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC)

(1) Experimental equipment

The device comprises a 96-well plate, a test tube, a 5uL-50uL micropipette, a 100uL-1000uL micropipette, a matched gun head, a 10mL centrifugal tube, a Mach turbidimetric tube, an ultra-clean workbench, an autoclave, a forced air drying box, a biochemical incubator, a MH broth culture medium, a PBS buffer solution and sterile water.

(2) Experimental methods

The method comprises the following steps: placing the prepared MH culture medium, PBS buffer solution, sterile water, a matched gun head and a centrifuge tube into an autoclave for sterilization, and sterilizing for 25min at 121 ℃. The test tubes are put into a forced air drying oven for sterilization at 170 ℃ for 2 h. And (4) carrying out ultraviolet sterilization on the 96-well plate on an ultra-clean workbench for more than 30 min.

Step two: taking out a strain to be tested from a refrigerator, activating for 0.5h, adding 5mL PBS buffer solution to the slant, slightly shaking the slant for 80 times on the palm to completely wash down the strain, and pouring the strain into a test tube to be gently shaken uniformly; sucking 1mL of bacterial liquid, adding the bacterial liquid into 4mL of PBS buffer solution, sucking 1mL of diluted bacterial suspension, sequentially carrying out 5-fold gradient dilution, and selecting 10⁸Bacterial suspension of about CFU/mL or 10⁶CFU/mL spore suspension (compare 0.5 McLeod turbiditube), ten times serial dilution of the selected bacterial suspension, and selecting the second test tube (i.e. concentration of about 10)⁶Bacterial suspension of about CFU/mL or 10⁴CFU/mL spore suspension), 0.5mL is sucked and added into 4.5mL MH culture medium, and the mixture is blown evenly by a gun head for standby.

Step three: preparing SU3327 medicinal liquid with concentration of 10.5 mg/ml by using dimethyl sulfoxide; SU3327 liquid medicine, bacterial liquid and MH broth culture medium are added into a 96-well culture plate by using a test tube double dilution method for overnight culture, and liquid medicine groups with different concentrations are all three in parallel, so that the reliability of experimental data is ensured. And (3) placing the 96-well culture plate in an incubator at 37 ℃ for incubation for 16h, and observing the turbid condition of the liquid in the wells, wherein the minimum concentration capable of completely inhibiting the growth of bacteria is the MIC value. After 100. mu.l of the culture was taken out from each well and cultured overnight in an incubator at 37 ℃ in a dilution-coated solid medium, colony growth was observed, and the minimum concentration at which the bacteria were completely killed was MBC value.

The experimental results of the above experiment are shown in table 1, and as shown in table 1, the bacteriostatic effect and bactericidal effect of SU3327 on the strains used in the experiment are better than those of gentamicin sulfate.

It should be noted that:

1. the two-fold dilution method for determining the MIC value of the bacteria can be replaced by a paper diffusion method, namely, a paper piece which is stained with liquid medicine with certain concentration is placed on a bacteria culture medium, and the inhibition zone formed between a bacteria growth area on the culture medium and the paper piece is measured to determine the drug sensitive MIC value of the bacteria;

2. the above MH broth medium may be replaced with other media such as agar medium;

3. the double dilution method for determining the MBC value can be replaced by the intrabasal inoculation method.

TABLE 1 MIC and MBC values of SU3327 and gentamicin sulfate for different strains

As shown in the table 1, the bacteriostatic effect and bactericidal effect of SU3327 on gram-negative pathogenic bacteria and gram-positive pathogenic bacteria are better than those of gentamicin sulfate.

Although the present invention has been described with reference to the preferred embodiments, it is not intended to be limited thereto, and variations and modifications may be made by those skilled in the art without departing from the spirit and scope of the present invention.

Claims

The C-JUN N-terminal kinase inhibitor SU3327 has antibiotic use.
2. The use of the C-JUN N-terminal kinase inhibitor SU3327 according to claim 1, characterized in that: the C-JUN N-terminal kinase inhibitor SU3327 is used for killing and inhibiting bacteria of different animal and human species.
3. The use of the C-JUN N-terminal kinase inhibitor SU3327 according to claim 2, characterized in that: the C-JUN N-terminal kinase inhibitor SU3327 is used for killing and inhibiting bacteria of a strain obtained by separating human and/or animal bodies or clinically separating the strain from infected persons and then culturing the strain.
4. The use of the C-JUN N-terminal kinase inhibitor SU3327 according to claim 1 or 2, characterized in that: the C-JUN N-terminal kinase inhibitor SU3327 is used for inhibiting and killing pseudomonas aeruginosa.
5. The use of the C-JUN N-terminal kinase inhibitor SU3327 according to claim 1 or 2, characterized in that: the C-JUN N-terminal kinase inhibitor SU3327 is used for inhibiting and killing staphylococcus aureus.
6. The use of the C-JUN N-terminal kinase inhibitor SU3327 according to claim 1 or 2, characterized in that: the C-JUN N-terminal kinase inhibitor SU3327 is used for inhibiting and killing bacillus subtilis.
7. The use of the C-JUN N-terminal kinase inhibitor SU3327 according to claim 1 or 2, characterized in that: the C-JUN N-terminal kinase inhibitor SU3327 is used for inhibiting and killing Escherichia coli.
8. The use of the C-JUN N-terminal kinase inhibitor SU3327 according to claim 1 or 2, characterized in that: the C-JUN N-terminal kinase inhibitor SU3327 is used for inhibiting and killing Candida albicans.
9. The use of the C-JUN N-terminal kinase inhibitor SU3327 according to claim 1 or 2, characterized in that: the C-JUN N-terminal kinase inhibitor SU3327 is used for inhibiting and killing acinetobacter baumannii.
10. The use of the C-JUN N-terminal kinase inhibitor SU3327 according to claim 1 or 2, characterized in that: the C-JUN N-terminal kinase inhibitor SU3327 is used for inhibiting and killing clostridium sporogenes.