CN116115664A - Coli antibacterial compound for inhibiting simultaneous carrying of multiple new Deril metal-beta-lactamase and application thereof - Google Patents
Coli antibacterial compound for inhibiting simultaneous carrying of multiple new Deril metal-beta-lactamase and application thereof Download PDFInfo
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- CN116115664A CN116115664A CN202310110432.7A CN202310110432A CN116115664A CN 116115664 A CN116115664 A CN 116115664A CN 202310110432 A CN202310110432 A CN 202310110432A CN 116115664 A CN116115664 A CN 116115664A
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- beta
- lactamase
- hypericum perforatum
- extract
- antibacterial
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Abstract
The invention discloses a natural plant-derived Hypericum perforatum antibacterial extract and application thereof, which can be used as an escherichia coli antibacterial compound carrying novel Derilieri metal-beta-lactamase-NDM 4, NDM5 and beta-lactamase TEM, ampC and application thereof, and is used for treating hydrocarbon mould enase drug-resistant bacteria infection. The antibacterial circle of the Hypericum perforatum extract on the escherichia coli carrying the novel Derileis metal-beta-lactamase-NDM 4, NDM5 and beta-lactamase TEM and AmpC is 30mm, and the minimum sterilization concentration is 0.01mg/mL. This will be helpful in the treatment of super-and drug-resistant bacterial infectious diseases caused by neodril metallo-beta-lactamase and beta-lactamase. The Hypericum perforatum has the advantages of convenient source, capability of being obtained and prepared in large quantity and good pharmaceutical application prospect.
Description
Technical Field
The invention relates to the field of biotechnology, in particular to application of a Hypericum perforatum extract from natural plants as a novel New Derildimetal-beta-lactamase bacteriostatic agent in the field of antibiosis.
Background
Coli is a bacterial Disease commonly occurring in pig farms and can cause a number of diseases in pig flocks, including neonatal diarrhea, weaned diarrhea/weaned bowel colibacillosis (Postweaning Enterotoxigenic e.coli, PWD), oedema/bowel oedema/visceral oedema (Edema Disease e.coli, ED), sepsis, multiple serositis, escherichia coli mastitis (coloriormMastis e.coli, CM), and urinary tract infections (Nonspecific Urinary Tract Infection e.coli, UTI), among others. Diarrhea caused by E.coli can lead to serious economic losses. Coli-based PWD and ED may occur simultaneously in one outbreak or in the same herd of pigs, or may occur independently. There is a local popularity in many pig farm PWDs. In the case of sow mastitis, the isolated pathogenic bacteria are mainly E.coli and E.coli groups. In addition, certain E.coli, especially O157: H7 serotype, O26 serotype and other non-O157 E.coli, may be scattered in the gut and feces of pigs and co-affected by humans and animals.
In recent years, a relatively serious escherichia coli infection is found in clinical cases, the severe escherichia coli infection is mainly represented by sudden death or severe diarrhea occurring 2-3 weeks after weaning, and escherichia coli susceptibility tests separated from the cases find that the escherichia coli infection has strong drug resistance and can resist various antibiotics. Overuse and abuse of antibiotics are the root cause of bacterial resistance, and multiple resistant strains and super resistant strains are clinically developed. The production of inactivated or modified enzymes is the primary mechanism of resistance development in E.coli, whereas the enzymes carbapenemase or beta-lactamase are the most abundant of these enzymes. To date, bacterial-produced beta-lactamases have been nearly 500. The most important enzymes that clinically lead to the resistance of gram-negative bacteria to carbapenems or beta-lactam antibiotics are the ultra-broad spectrum beta-lactamases (Extended spectrum beta-kactanases, ESBLs), the AmpC beta-lactamases (AmpC beta-lactamases, ampCs) and the neodri metallo beta-lactamases (New Delhi metallo beta-lactamases, NDM). The NDM genes mainly comprise tens of subtypes of NDM-1 to NDM-50, wherein the prevalence of the genes of E.coli NDM4 and NDM-5 is increasing. TEM type ESBLs are a series of enzyme proteins formed by mutating a coding gene of broad-spectrum penicillin enzyme TEM-1 or TEM-2 by 1-4 amino acids, and are the most abundant beta-lactamase at present. AmpC beta-lactamase is gram-negative bacteria produced by not being clavulanic acid inhibited "serine" cephalosporin enzyme, it can be both chromosome-mediated and plasmid-mediated, the preferred hydrolysis substrate for such enzyme is cephalosporin, belonging to class I of the Bush-J-M class, class C of the Ambler class.
With the wide clinical application of carbapenem and beta-lactam antibiotics, the escherichia coli has drug resistance to carbapenem and beta-lactam antibiotics, and the drug resistance tends to be gradually increased, in particular to the emergence of escherichia coli carrying NDM genes. The abuse of antibiotics has led to the massive emergence and global spread of multi-drug resistant bacteria and "superbacteria". Such bacteria are very resistant to almost all known antibiotics. In recent years, the types and numbers of superbacteria clinically isolated are increasing, and the main bacteria that have been detected to produce NDM genes include Klebsiella pneumoniae, morganella morganii, escherichia coli, pseudomonas aeruginosa, klebsiella pneumoniae, enterobacter cloacae, citrobacter, acinetobacter baumannii, klebsiella oxytoca, pseudomonas aeruginosa, proteus, nonfermentation gram-negative bacteria, salmonella, citrobacter freundii, providencia and the like. More NDM-1 was clinically isolated, followed by successive reports of NDM-2, NDM-3, NDM-4, NDM-5, NDM-6, etc. The mechanism by which NDM and its subtype super bacteria develop resistance is that resistant strains produce new deli metallo beta-lactamase (NDM), an important carbapenemase enzyme capable of hydrolysing carbapenem antibiotics. NDM-4 was first found in e.coli and was resistant to imipenem and ertapenem. There are studies reporting that carrying blaNDM-4 is on IncFIA and IncF type plasmids. The currently popular strains are E.coli, klebsiella pneumoniae and Enterobacter cloacae. NDM-5 was first reported in 2011 in the united kingdom, and this strain exhibited a multi-drug resistant phenotype, isolated from a human strain of escherichia coli. NDM-5 has better enzyme activity than NDM-1, and can hydrolyze cefotaxime, ceftiofur and ceftazidime with high efficiency. BlaNDM-5 has been found on plasmids such as IncF, incFII, incN and IncX3, and the major epidemic strains are Shigella, klebsiella pneumoniae and Escherichia coli, and epidemic areas are prevalent in multiple countries worldwide.
Because the antibiotics are applied to livestock breeding, huge hidden danger and food safety hidden danger are brought while animal epidemic diseases are prevented and controlled and animal production performance is improved, the drug resistance of animal clinical bacteria is enhanced, multiple drug resistant bacteria and super drug resistant bacteria appear, the animal clinical common antibiotics are resistant, and the dilemma of drug-free selection appears. The plant extract has antibacterial, antiviral and anti-inflammatory functions, is not easy to generate drug resistance, has the effects of improving animal immunity and animal production performance, and is the optimal drug for replacing antibiotics.
Hypericum perforatum (Hypericum perforatum Linn), also known as Hypericum perforatum, st. John's grass, is a perennial herb of Hypericum genus of Guttiferae, and is the most widely distributed natural plant in the world. The Hypericum perforatum extract is used as a purely natural plant medicine, has definite curative effect in the aspects of treating mild-moderate depression, anxiety disorder, combined treatment of insomnia, alcohol dependence syndrome, benign paroxysmal positional dizziness, cardiovascular neurosis, digestive system diseases and the like, and has good safety and tolerance. However, hypericum perforatum extract as novel New Deril Metal inhibitor the novel use of beta-lactamase bacteriostats has not been reported.
In 2002, li Hong et al reported that the total Hypericum perforatum extract has antibacterial and bactericidal effects on Staphylococcus aureus (gram positive bacteria), and its antibacterial or bactericidal effects are related to its concentration.
In 2002, li Hong et al reported the antibacterial effect of the total extract of Hypericum perforatum on Corynebacterium glutamicum (gram positive bacteria) and Alcaligenes faecalis (gram negative bacteria), and the results indicate that the total extract has strong antibacterial and bactericidal effects on Corynebacterium glutamicum; the bacillus caldus has no bactericidal effect but has bacteriostatic effect, and the effect intensity is related to the concentration.
In 2004, li Hong et al reported the antibacterial effect of Hypericum perforatum total extract on Bacillus thuringiensis 7216 (gram positive bacteria), and the results indicate that the extract has strong antibacterial and bactericidal effects on 7216, and that these two effects are related to their concentrations.
In 2004, li Hong et al reported the antibacterial effect of Hypericum perforatum total extract on Pseudomonas aeruginosa (gram positive bacteria), and the results indicate that the extract has only weak antibacterial effect on Pseudomonas aeruginosa and no bactericidal effect.
In 2005, li Hong et al reported the antibacterial effect of Hypericum perforatum total extract on Brevibacterium flavum (gram positive bacteria), and the results indicate that the extract has strong antibacterial and bactericidal effects on the bacteria, and the two effects are related to the concentration thereof.
In 2006, li Hong et al reported the antibacterial effect of Hypericum perforatum total extract on 11 pathogenic bacteria (gram positive bacteria), and the results indicate that the extract has strong antibacterial and bactericidal effects on all strains, and the results suggest that the extract can be used for controlling infections caused by gram positive strains which are clinically resistant to various antibiotics.
In 2007, li Hong and the like report the antibacterial effect of the Hypericum perforatum total extract on pathogenic bacteria, and the result shows that the extract has strong antibacterial and bactericidal effects on tested gram-positive bacterial strains, weak antibacterial effects on a few gram-negative bacterial strains, and no bactericidal effect.
As known in the prior art, the Hypericum perforatum total extract has strong antibacterial and bactericidal effects on gram-positive bacterial strains. However, the new application of Hypericum perforatum as a gram negative escherichia coli bacteriostatic agent for inhibiting novel Derildimetal-beta-lactamase is not reported yet.
Disclosure of Invention
The invention aims at the application of Hypericum perforatum extract as a bactericide of escherichia coli carrying novel metal beta-lactamase in the antibacterial field.
The application of herba Hyperici perforati extract in preparing antibacterial and bactericide of Escherichia coli carrying metal beta-lactamase is provided.
The metallo-beta-lactamase is at least one or more of NDM4 type beta-lactamase, NDM5 type beta-lactamase, TEM type beta-lactamase and AmpC type beta-lactamase.
Use of Hypericum perforatum extract in the preparation of antibacterial and bactericidal agent for treating Escherichia coli carrying or simultaneously carrying metallo-beta-lactamase-NDM 4, NDM5, TEM and AmpC.
An antibacterial compound comprises herba Hyperici perforati extract, wherein the content of total hypericin in herba Hyperici perforati extract is 0.3%.
The Hypericum perforatum extract inhibits escherichia coli carrying metallo-beta-lactamase, wherein the dosage of the Hypericum perforatum extract is greater than or equal to 10 mug/mL.
Hypericum perforatum extract contains hypericin and pharmaceutically acceptable drug carrier.
The medicinal preparation is powder, tablet, capsule, injection, etc.
Use of a compound for the preparation of a medicament for the treatment of an antibacterial agent.
Use of Hypericum perforatum extract for the treatment of E.coli infections carrying metallo-beta-lactamase.
The Hypericum perforatum extract is used for treating infection caused by escherichia coli carrying metallo-beta-lactamase-NDM 4, NDM5, TEM and AmpC simultaneously, and the dosage of the Hypericum perforatum extract is more than or equal to 10 mug/mL.
The invention has the advantages and innovation points that: the new application of the Hypericum perforatum extract is found, and the application of the bactericide for the escherichia coli carrying the novel metal beta-lactamase in the antibacterial field is realized. Experiments prove that the Hypericum perforatum extract has better antibacterial activity and bactericidal activity of metal-beta-lactamase (NDM 4, NDM5, TEM and AmpC), the diameter of a bacteriostasis circle is 30mm, and the minimum bactericidal concentration is 0.01mg/mL. Therefore, the Hypericum perforatum extract can be used as a candidate drug of a novel metal-beta-lactamase-producing bactericide, and has good pharmaceutical application prospect. Provides a new choice for clinically treating infectious diseases caused by hydrocarbon zymone gram-negative drug-resistant bacteria. Provides reference for the prevention and treatment of gram-negative multi-drug resistant bacteria and super-drug resistant bacteria in future, and lays a foundation for the reduction of resistance, the replacement of resistance and the cultivation without resistance.
Drawings
FIG.1 is a sample evolution analysis based on 16S rRNA, fig.1. Physical tree of16S rRNA genes.
FIG.2 is a HN2187 genome circle map, fig.2 colorimeter circles.
FIG.3 is a circle of pNDM4-HN2187 (Plasmid B), FIG.3pNDM4-HN2187 (Plasmid B) circle.
FIG. 4 is a pNDM4-HN2187 (PlasmidB) treelet analysis, FIG. 4Phylogenetic tree of. pNDM4-HN2187 (Plasmid B).
FIG.5 is a circle of pNDM5-HN2187 (plasmid A), FIG.5pNDM5-HN2187 (plasmid A) circle.
FIG. 6 is a graph of 20types of antibiotic resistance genes, FIG. 620 and FIG. types ofantibiotic resistance genes. Fig. 7 shows the result of the inhibition of HN2187 by Hypericum perforatum with zone 30mm,Fig 1Antibacterial results ofPerilla frutescens,the antibacterial zone was 30mm.
Detailed Description
The invention is further illustrated by the following examples, which are not intended to limit the invention in any way, and any modifications or alterations to the invention, which would be readily apparent to a person of ordinary skill in the art, without departing from the technical solutions of the invention, are intended to fall within the scope of the claims of the invention.
EXAMPLE 1 porcine E.coli whole genome sequencing analysis and drug susceptibility experiments carrying New Deril metallobeta-lactamase NDM4, NDM5 and beta-lactamase blaTEM and AmpC enzymes simultaneously
1.1 materials
1.1.1 Strain sources 1 multiple drug resistant strains separated and identified in 91 E.coli strains separated and identified in the clinical pig diarrhea cases in 2021 of the laboratory, drug resistant to imipenem, ertapenem, cefoxitin, penicillin and the like, and the strains are named as follows: escherichia coli is preserved in China general microbiological culture Collection center (CGMCC) with the address of North Xielu No. 1, 3 in the Korean area of Beijing, and the preservation date of 2021, 8 months and 26 days, and the preservation number of CGMCC No.23303. The escherichia coli carrying NDM, blaTEM, ampC genes is determined through PCR detection and whole genome sequencing and is preserved at-80 ℃ for standby.
1.1.2 reagents and equipment imipenem, ertapenem and cefoxitin were purchased from Shanghai source leaf biotechnology Co., ltd; the drug sensitive paper such as penicillin G, cefotaxime, ceftizoxime, ceftriaxone, cefoperazone, ampicillin, amoxicillin, oxacillin, cefalotin, cefprozil, cefadroxil, cefaclor, cefuroxime, cloxacillin, ceftazidime, ceftiofur, ceftizoxime, cefquinome and the like is purchased from Beijing Tiantan drug biotechnology development Co, and the reagents required for primer synthesis and molecular experiments are purchased from Bao Ri doctor technology (Beijing) Co.
1.2 method
1.2.1 sample evolution analysis based on 16S rRNA A phylogenetic tree was constructed by selecting 19 strains closest at the species level based on the 16S rRNA sequence by comparison with a local database, and by MEGA 6.0 software selection NJ (Neighbor-training) method.
1.2.2 screening of drug resistance genes carbapenemases NDM4, NDM5 and beta-lactamase blaTEM and AmpC enzyme drug resistance genes were reviewed according to the literature, and the gene primer information is shown in Table 1.
TABLE 1NDM-5 drug resistance Gene primer sequence Table 1Primer sequence ofNDM-5resistance gene
1.2.3 drug susceptibility testing drug resistance phenotyping was performed using a drug susceptibility paper sheet method.
1.2.4 genome sequencing and analysis full genome sequencing was performed using a second-generation + third-generation sequencing approach, i.e., illumina Hiseq + pacbrio.
1.3 results and analysis
1.3.1 sample evolution analysis based on 16S rRNA
HN2187 belongs to one branch with Escherichia coli by comparison with phylogenetic tree construction, and is Escherichia coli (FIG. 1). FIG.1 is a sample evolution analysis based on 16S rRNA, fig.1. Phytologic tree of16S rRNA genes.
1.3.2 drug sensitivity of Escherichia coli HN2187
Drug susceptibility tests were performed on clinically usual beta-lactam antibiotics and carbapenem antibiotics, and the drug susceptibility results are shown in Table 2. The strain is totally resistant to carbapenem and beta-lactam antibiotics commonly used in clinic.
TABLE 2 drug sensitivity test results Table 2The results ofthe drug susceptibility testing
Note that: r, drug resistance
Note：R，drug resistance
1.3.3 genomic features of E.coli HN2187 genome circles were mapped by whole genome sequencing (FIG. 2). The genomic map of E.coli HN2187 contained 5270080bp with a G+C (%) content of 50.63%. Plasmid identification was performed on the bacterial genome assembly results using plasmid flow (https:// github. Com/smaegol/plasmid flow) software, and plasmid annotation was performed using BLAST (https:// BLAST. Ncbi. Lm. Gov/BLAST. Cgi) software and PLSDB database (https:// ccb-micro. Cs. Uni-saarland. De/PLSDB /). Analysis showed that HN2187 contained 2 plasmids (Plasmid A, plasmid B), 2 types, type IncHI and type IncFIC (FII), respectively. FIG.2 is a HN2187 genome circle map, fig.2 colorimeter circles
1.3.4pNDM4-HN2187 and blaTEM-HN2187 (Plasmid B) Plasmid analysis pNDM4-HN2187 (Plasmid B) was 159194bp in length and 50.86% GC content (FIG. 3). FIG.3 is a circle of pNDM4-HN2187 (Plasmid B), FIG.3pNDM4-HN2187 (Plasmid B) circle.
And analysis shows that the plasmid contains 2 drug resistance genes, namely bla NDM-4 And bla TEM-1 Which code for the products NDM-4 and bla TEM-1 Mainly resistant to carbapenem antibiotics, cefoxitin and beta-lactam antibiotics. Bla TEM-1 The gene ID is pB_gene0140, which is located at Plasmid B135324-136184,
the size is 861bp. PlasFlow analysis showed that this plasmid type was IncFIC (FII). BLAST alignment of the complete sequence of pNDM4-HN2187 (Plasmid B) with the GenBank database revealed that this Plasmid was matched with the 1 Plasmid recently reported (p-NDM-4,AccessionNo CP050167.1) (51% coverage and 99.99% sequence similarity, this Plasmid was derived from E.coli isolated from hong Kong in China). To further determine the genotype of pNDM4-HN2187 (Plasmid B), the 9 genes were phylogenetic tree-typed with MEGA software, and the typing results were shown (FIG. 4), FIG. 4 is
pNDM4-HN2187 (Plasmid dB) treelet analysis, FIG. 4Phylogenetic tree of pNDM4-HN2187 (Plasmid B). The genotype of pNDM4-HN2187 (Plasmid B) was matched to (p-NDM-4,AccessionNo CP050167.1) (51% coverage and 99.99% sequence similarity, a Plasmid derived from E.coli isolated from hong Kong China) and was recently pooled into the same branch.
1.3.5pNDM5-HN2187 (PlasmidA) plasmid analysis pNDM5-HN2187 (PlasmidA) was 216628bp in length and 45.6% GC content (FIG. 5). FIG.5 is a circle of pNDM5-HN2187 (plasmid A), FIG.5pNDM5-HN2187 (plasmid A) circle. And analysis shows that the plasmid only contains 1 drug resistance gene bla NDM5 The coded product NDM is mainly involved in the tolerance to carbapenem antibiotics. Bla NDM5 Gene ID pA_gene0009, position PlasmidA: compplement (5743-6555). PlasFlow analysis showed that the plasmid type was of the IncHI type. BLAST comparison of the complete sequence of the pNDM5-HN2187 (Plasmid B) gene with the GenBank database revealed that this Plasmid had a high homology to the 68 recently reported NDM5 plasmids, reaching 100% (Table 3).
TABLE 3 partial plasmid with higher homology to pNDM5-HN2187 (plasmid) NDM5 gene
Table 3Partial plasmid with high homology to PNDM5-HN2187(plasmid)
NDM5 gene
1.3.6AmpC chromosomal Gene analysis the AmpC gene was located on the chromosome of HN2187, with two AmpC genes located at different positions on the chromosome, with gene IDs gene1306 and gene4188, respectively, at positions (1357262-1358566) and 4373156-4374289 on the chromosome, respectively, of sizes 1305bp and 1134bp, respectively.
1.3.7 E.coli HN2187 resistance Gene prediction comprehensive analysis of the database of antibiotic resistance genes showed that (ComprehensiveAntibiotic Resistance Database, CARD) HN2187 contained 20types (269) of antibiotic resistance genes (FIG. 6), 620types of antibiotic resistance genes, FIG. 620types ofantibiotic resistance genes. Including macrolides, tetracyclines, fluoroquinolones, penicillanes, cephalosporins, phenylpropanoids, cephalosporins, aminoglycosides, carbapenems, rifamycins, peptides, monoamines, enzymes, aminocoumarins, glycylcyclines, glycopeptides, triclosan, acridine dyes, lincomides, pleuromutilins, and others.
1.4 isolation of 91 E.coli (E.coli) from 2021 (1-12 months) clinical cases of porcine diarrhea 1 E.coli carrying both the New Derilmetal beta-lactamase NDM4, NDM5 and the beta-lactamase blaTEM, ampC enzymes was identified. 11 antibiotic drug susceptibility tests were performed, and the results showed that HN2187 strain was resistant to both. The strain produces B-type enzyme, carries NDM4, NDM5, blaTEM and AmpC enzyme drug resistance genes, wherein blaNDM-4 is positioned on Plasmid pNDM4-HN2187 (Plasmid B), blaNDM-5 is positioned on Plasmid pNDM5-HN2187 (Plasmid A), blaTEM1 is positioned on Plasmid pTEM1-HN2187 (Plasmid B), and AmpC enzyme drug resistance genes are positioned on chromosomes, and mediate blaNDM-4, blaNDM-5, blaTEM and AmpC drug resistance genes respectively, and are important vectors for drug resistant bacteria transmission.
Example 2 Hypericum perforatum extract extraction process
(1) Crushing Hypericum perforatum, and then using a feed liquid ratio of 1:10, soaking the mixture in 75 to 80 percent of ethanol for 2 hours, ultrasonically extracting the mixture for 2 hours by using an ultrasonic extraction method, filtering the mixture, and collecting supernatant;
(2) Filter residue was purified by 1: extracting with water at 60deg.C for 1 hr, filtering, and collecting supernatant;
(3) Mixing the two supernatants, concentrating, recovering ethanol, and concentrating to obtain paste;
(4) Adding pharmaceutically acceptable adjuvants, concocting into paste, spraying powder at high temperature, measuring total hypericin content, and concocting with adjuvants to obtain hypericin content of 0.3%.
Example 3 Hypericum perforatum extract bacteriostasis experiment
The method of measuring the inhibition zone is adopted. Taking Hypericum perforatum extract, adjusting the concentration to 1g/mL, taking 100 mu L to do bacteriostasis experiment, respectively doing three repetitions, and taking the average value. The diameter of the inhibition zone is 30mm (see figure 7). As can be seen from FIG.1, the Hypericum perforatum extract has a good antibacterial effect on gram-negative bacteria and carries Escherichia coli carrying NDM4 type beta-lactamase, NDM5 type beta-lactamase, TEM type beta-lactamase and AmpC type beta-lactamase, and can be used as a bacteriostatic agent for the gram-negative bacteria.
Test example 4 Hypericum perforatum extract sterilization experiment
The method of measuring the minimum sterilization concentration is adopted. The initial concentration of Hypericum perforatum was 1g/mL, then 10-fold diluted, 10 serial dilutions were made. Three replicates were performed separately and averaged. The minimum sterilization concentration of the Hypericum perforatum is 0.01mg/mL, which shows that the Hypericum perforatum has better antibacterial effect on gram-negative bacteria and simultaneously carries escherichia coli carrying NDM4 type beta-lactamase, NDM5 type beta-lactamase, TEM type beta-lactamase and AmpC type beta-lactamase, and can be used as a bactericide of the gram-negative bacteria.
Test example 5 toxicity counteracting and therapeutic experiments
Preparing coliform suspension with NDM4 type beta-lactamase, NDM5 type beta-lactamase, TEM type beta-lactamase and AmpC type beta-lactamase to obtain bacterial suspension with bacterial concentration of1×10 9 . The weaned pigs with 20 heads and consistent body weight and size are selected and divided into an experimental group and a control group, wherein each head of the weaned pigs in the experimental group attacks 0.5mL of escherichia coli, the control group is injected with 0.5mL of physiological saline for 24 hours and clinical symptoms appear, the experimental group is treated by taking Hypericum perforatum extract (20 g per day), the control group is treated by antibiotics, and the survival rate and the death rate are counted after 3 d. The experimental group treated by the Hypericum perforatum had all survived, the survival rate was 100%, the control group treated by the antibiotics had 8 deaths, 2 survived, and the survival rate was 20%. The Hypericum perforatum extract has better treatment effect on the infection of the cases of the escherichia coli carrying the NDM4 type beta-lactamase, the NDM5 type beta-lactamase, the TEM type beta-lactamase and the AmpC type beta-lactamase, and can be used as a medicament for treating the escherichia coli.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (10)
1. The application of herba Hyperici perforati extract in preparing antibacterial and bactericide of Escherichia coli carrying metal beta-lactamase is provided.
2. The use according to claim 1, wherein the metallo-beta-lactamase is at least one or more of NDM4 beta-lactamase, NDM5 beta-lactamase, TEM beta-lactamase and AmpC beta-lactamase carried simultaneously.
3. Use of the Hypericum perforatum extract according to claim 1 for the preparation of an E.coli antibacterial and bactericidal agent carrying or simultaneously carrying metallo-beta-lactamase-NDM 4, NDM5, TEM and AmpC.
4. An antibacterial compound comprising Hypericum perforatum extract, wherein Hypericum perforatum extract has a total hypericin content of 0.3%.
5. The antibacterial compound according to claim 4, wherein the extract of Hypericum perforatum inhibits Escherichia coli harboring metallo-beta-lactamase, and wherein the amount of Hypericum perforatum extract is 10 μg/mL or more.
6. The antibacterial compound of claim 4, wherein the Hypericum perforatum extract comprises hypericin and a pharmaceutically acceptable pharmaceutical carrier thereof.
7. The antibacterial compound of claim 6, wherein the pharmaceutical formulation is a powder, tablet, capsule, injection, or the like.
8. Use of a compound according to claim 6 or 7 for the preparation of a medicament for the treatment of an antibacterial agent.
9. Use of Hypericum perforatum extract for the treatment of E.coli infections carrying metallo-beta-lactamase.
10. The use according to claim 9, wherein the Hypericum perforatum extract is used for the treatment of infections caused by E.coli harboring metallo-beta-lactamase-NDM 4, NDM5, TEM and AmpC simultaneously, in an amount of greater than or equal to 10 μg/mL.
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1689596A (en) * | 2004-04-21 | 2005-11-02 | 上海市中药研究所 | Hypericum perforatum extract and its preparation process |
| CN101797374A (en) * | 2010-03-24 | 2010-08-11 | 天津生机集团股份有限公司 | Compound minocycline hydrochloride soluble powder for treating respiratory diseases of poultry |
| CN104474030A (en) * | 2014-11-21 | 2015-04-01 | 河南省科高植物天然产物开发工程技术有限公司 | Glucoside composition from forsythia suspense leaves and application thereof |
| WO2015157618A1 (en) * | 2014-04-11 | 2015-10-15 | The Texas A&M University System | Novel inhibitors of the new delhi metallo beta lactamase (ndm-1) |
| CN105748689A (en) * | 2014-12-16 | 2016-07-13 | 天津德瑞祥科技有限公司 | Traditional Chinese veterinary medicinal compound composition for preventing and treating serious Escherichia coli infection of livestock and poultry, and preparation method thereof |
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| CN114272298A (en) * | 2021-11-12 | 2022-04-05 | 河南省农业科学院畜牧兽医研究所 | Willow bark extract, compound preparation and application thereof |
| CN115120584A (en) * | 2021-03-26 | 2022-09-30 | 中国医学科学院医药生物技术研究所 | New Delhi metallo-beta-lactamase-1 inhibitors |
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2023
- 2023-02-14 CN CN202310110432.7A patent/CN116115664A/en active Pending
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| CN1689596A (en) * | 2004-04-21 | 2005-11-02 | 上海市中药研究所 | Hypericum perforatum extract and its preparation process |
| CN101797374A (en) * | 2010-03-24 | 2010-08-11 | 天津生机集团股份有限公司 | Compound minocycline hydrochloride soluble powder for treating respiratory diseases of poultry |
| WO2015157618A1 (en) * | 2014-04-11 | 2015-10-15 | The Texas A&M University System | Novel inhibitors of the new delhi metallo beta lactamase (ndm-1) |
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| WO2016169491A1 (en) * | 2015-04-23 | 2016-10-27 | 富力 | Use of forsythin, derivatives thereof and composition of forsythin/forsythiaside in the preparation of drug for treating bacterial infections |
| US20200093814A1 (en) * | 2016-12-21 | 2020-03-26 | Aicuris Gmbh & Co. Kg | Combination therapy with amidine substituted beta-lactam compounds and beta-lactamase inhibitors for infections with antibiotic resistant bacterial strains |
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| OKAY NAZLI: "Antimicrobial and antibiofilm activity of polyurethane/Hypericum perforatum extract (PHPE) composite", BIOORGANIC CHEMISTRY, vol. 82, pages 224 - 228 * |
| SEBASTIÃO D. SANTOS-FILHO: "Infl uence of an aqueous extract of Hypericum perforatum (Hypericin) on the survival of Escherichia coli AB1157 and on the electrophoretic mobility of pBSK plasmid DNA", REVISTA BRASILEIRA DE FARMACOGNOSIA BRAZILIAN JOURNAL OF PHARMACOGNOSY, vol. 18, no. 3, pages 326 - 330 * |
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