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Application of L-rhamnose antibody in preparation of medicine for preventing and/or treating drug-resistant bacterial infection

A technology of drug-resistant bacteria and rhamnose, which is applied in the field of medicine, can solve the problems of staying in the experimental research stage, the difficulty of developing antibiotics, and the weak and incapable of bacterial invasion, so as to achieve strong inhibition and killing ability and improve L-rhamnose antibody Level, good killing effect

Pending Publication Date: 2021-11-16
SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for some super-resistant bacteria, there is often no drug available
In addition, people have conducted a lot of research on drug-resistant bacteria, and they are mainly expected to overcome drug-resistant bacterial infections in the following ways: 1) Develop new antibiotics directly targeting drug-resistant bacteria, but the speed of bacterial drug resistance far exceeds the development of new antibiotics Speed, and the development of antibiotics is becoming more and more difficult
2) To overcome the mechanism of drug resistance and restore the susceptibility of bacteria to antibacterial drugs, for example, for the β-lactamase produced by drug-resistant bacteria, research on synthetase inhibitors, the combined use of enzyme inhibitors and antibiotics, in overcoming bacterial resistance At the same time, it exerts the bactericidal effect of antibiotics, but so far, only the compound application of β-lactamase inhibitors and penicillins (or cephalosporins) is available clinically, and many other drug resistance mechanisms cannot be overcome
3) Alternative products to antibiotics, such as antibacterial peptides, phages, etc., but most of them remain in the experimental research stage and are still far away from clinical application
Weak natural antibody levels to L-rhamnose may not provide effective protection against this part of the bacterial attack

Method used

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  • Application of L-rhamnose antibody in preparation of medicine for preventing and/or treating drug-resistant bacterial infection
  • Application of L-rhamnose antibody in preparation of medicine for preventing and/or treating drug-resistant bacterial infection
  • Application of L-rhamnose antibody in preparation of medicine for preventing and/or treating drug-resistant bacterial infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] The preparation method steps of human source L-rhamnose natural antibody are as follows:

[0085] (1) Synthesis of L-rhamnose-NH 2

[0086] 1 mol of L-rhamnose was protected by acetyl group and 1.1 mol of thiophenol was glycosylated to obtain intermediate product 1, and then 1 mol of intermediate product 1 was connected with 2 mol of 3-azidopropanol to obtain intermediate product 2. Product 2 was protected by deacetylation and reduced to obtain L-rhamnose-NH 2 ;

[0087]

[0088] (2) Preparation of L-rhamnose affinity chromatography column

[0089] Take the NHS-activated agarose gel column, wash the column with 10 column volumes of pre-cooled, 1mM HCl, and wash the column with 5 column volumes of 0.2M NaHCO 3 , 0.5M NaCl solution (pH=8.3) to equilibrate the column; add 2 mL of L-rhamnose-NH with a concentration of 10 mg / mL 2 After that, mix evenly, react with slow shaking at room temperature for 4 hours, continue to add 3 column volumes of 0.1M Tris-HCl solution...

Embodiment 2

[0094] Example 2 Preparation of mouse L-rhamnose monoclonal antibody

[0095] The preparation method step of mouse source L-rhamnose monoclonal antibody is as follows:

[0096] (1) Synthesis of L-rhamnose-NHS

[0097] 1mol of L-rhamnose-NH 2 Reaction with 1.1 mol of succinyl gave intermediate 3, which was activated by 1.1 mol of 2-succinimidyl-1,1,3,3-tetramethylurea tetrafluoroborate to give L-rhamnose-NHS;

[0098]

[0099] (2) Preparation of L-rhamnose-OVA

[0100] Dissolve 10mg of L-rhamnose-NHS in 1mL 3×PBS buffer (pH=7.4); another 10mg of OVA was dissolved in 1mL 3×PBS buffer (pH=7.4); Stir for 1 hour; transfer the above reaction system to a 3kDa ultrafiltration tube, centrifuge at 4°C for 30 minutes at 3,500 rpm, add an appropriate amount of 1×PBS buffer in the tube, and centrifuge at 4°C for 30 minutes at 3,500 rpm, repeat 3 times, The solution obtained in the casing is L-rhamnose-OVA;

[0101] (3) Preparation of mouse splenocyte suspension

[0102] Take five...

Embodiment 3

[0110] Example 3 Bactericidal effect test of human L-rhamnose natural antibody

[0111] Bacterial lipopolysaccharide extraction and SDS-PAGE / silver staining detection, the specific method steps are as follows:

[0112] (1) Extracting bacterial lipopolysaccharide: inoculate 5 μL of glycerol bacteria into 5 mL of LB liquid medium, and cultivate overnight at 37° C. at 220 rpm; centrifuge to collect overnight cultured cells, and extract lipopolysaccharide with a lipopolysaccharide extraction kit; Glycerol bacteria are Escherichia coli or Pseudomonas aeruginosa;

[0113] (2) SDS-PAGE / silver staining detection: Take 5 μL of the extracted bacterial lipopolysaccharide, add 1 μL 6×SDS sample buffer, mix well and boil for 6 minutes; the obtained sample is used for 12% SDS-PAGE electrophoresis; the electrophoresis is over After that, in fixative solution (35mL H 2 O, 15mL acetic acid, 50mL isopropanol), shake slowly for 2 hours, then wash the gel twice with 200mL 7.5% glacial acetic ac...

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Abstract

The invention relates to application of an L-rhamnose antibody in preparation of a drug for preventing and / or treating drug-resistant bacterial infection. The L-rhamnose antibody is used for specifically recognizing L-rhamnose, and the L-rhamnose antibody is a human source L-rhamnose natural antibody or a mouse source L-rhamnose monoclonal antibody; the drug-resistant bacteria particularly belong to drug-resistant bacteria of which the genome contains an L-rhamnose synthesis route. According to the application disclosed by the invention, L-rhamnose synthetic pathways exist in genomes of a considerable part of drug-resistant bacteria, so that L-rhamnose exists in polysaccharide structures on the surfaces of many drug-resistant bacteria, and L-rhamnose antibodies can recognize and be combined with the L-rhamnose structures on the surfaces of the drug-resistant bacteria; further, the compound has extremely strong inhibiting and killing capability on various drug-resistant bacteria including pseudomonas aeruginosa, escherichia coli and streptococcus pneumoniae, and has the advantages of broad spectrum and good killing effect.

Description

technical field [0001] The invention relates to the application of L-rhamnose antibody in the preparation of medicines for preventing and / or treating drug-resistant bacterial infections, and belongs to the technical field of medicine. Background technique [0002] Drug-resistant bacteria refer to bacteria that are resistant to corresponding antibiotics after long-term antibiotic selection. The long-term, widespread, and excessive use of antibiotics has caused some bacteria to mutate into drug-resistant bacteria. This drug resistance will not only be passed on to the next generation, but also acquired by other bacteria. [0003] Bacterial drug resistance has become a serious issue of common concern to the global medical community. At present, clinical methods for treating drug-resistant bacterial infections are very limited, mainly through sputum culture and / or drug susceptibility testing to adjust the type or dose of antibiotics used. However, for some super drug-resistant...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61K9/08A61K9/06A61K47/10A61P31/04C07K16/44C12N5/20
CPCC07K16/44A61K9/08A61K9/0019A61K9/06A61K47/10A61P31/04A61K2039/505Y02A50/30
Inventor 马中瑞张化杰
Owner SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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