CN105998153B - Ramulus Et Folium Pithecellobii Lucidi extract is preparing the application in anti-ESBLs-producing bacteria Escherichia coli drug - Google Patents
Ramulus Et Folium Pithecellobii Lucidi extract is preparing the application in anti-ESBLs-producing bacteria Escherichia coli drug Download PDFInfo
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- 239000000284 extract Substances 0.000 title claims abstract description 115
- 239000003814 drug Substances 0.000 title claims abstract description 66
- 241000588724 Escherichia coli Species 0.000 title claims abstract description 58
- 229940079593 drug Drugs 0.000 title abstract description 29
- 230000003115 biocidal effect Effects 0.000 claims abstract description 41
- 229960004821 amikacin Drugs 0.000 claims abstract description 39
- LKCWBDHBTVXHDL-RMDFUYIESA-N amikacin Chemical compound O([C@@H]1[C@@H](N)C[C@H]([C@@H]([C@H]1O)O[C@@H]1[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O1)O)NC(=O)[C@@H](O)CCN)[C@H]1O[C@H](CN)[C@@H](O)[C@H](O)[C@H]1O LKCWBDHBTVXHDL-RMDFUYIESA-N 0.000 claims abstract description 39
- 150000001875 compounds Chemical class 0.000 claims abstract description 32
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 138
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
- 238000000034 method Methods 0.000 claims description 20
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 5
- 241000282693 Cercopithecidae Species 0.000 claims description 2
- JEGUKCSWCFPDGT-UHFFFAOYSA-N h2o hydrate Chemical compound O.O JEGUKCSWCFPDGT-UHFFFAOYSA-N 0.000 claims description 2
- 230000000844 anti-bacterial effect Effects 0.000 abstract description 29
- 238000012360 testing method Methods 0.000 abstract description 24
- 230000002195 synergetic effect Effects 0.000 abstract description 14
- 206010059866 Drug resistance Diseases 0.000 abstract description 8
- 229940088710 antibiotic agent Drugs 0.000 abstract description 6
- 201000010099 disease Diseases 0.000 abstract description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 4
- 238000013459 approach Methods 0.000 abstract description 2
- 244000033373 Pithecellobium clypearia Species 0.000 description 79
- 241000894006 Bacteria Species 0.000 description 47
- 239000000469 ethanolic extract Substances 0.000 description 46
- 238000004519 manufacturing process Methods 0.000 description 23
- 230000000694 effects Effects 0.000 description 16
- 230000002401 inhibitory effect Effects 0.000 description 15
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- 230000008485 antagonism Effects 0.000 description 12
- 241000196324 Embryophyta Species 0.000 description 10
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- 238000004659 sterilization and disinfection Methods 0.000 description 10
- 239000000243 solution Substances 0.000 description 9
- 206010070834 Sensitisation Diseases 0.000 description 8
- 238000004500 asepsis Methods 0.000 description 8
- 238000010790 dilution Methods 0.000 description 8
- 239000012895 dilution Substances 0.000 description 8
- 238000005259 measurement Methods 0.000 description 8
- 230000008313 sensitization Effects 0.000 description 8
- 238000007619 statistical method Methods 0.000 description 8
- 239000000725 suspension Substances 0.000 description 8
- 235000019441 ethanol Nutrition 0.000 description 7
- 230000035945 sensitivity Effects 0.000 description 6
- 238000009826 distribution Methods 0.000 description 5
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 239000011248 coating agent Substances 0.000 description 4
- 238000000576 coating method Methods 0.000 description 4
- 230000000249 desinfective effect Effects 0.000 description 4
- 230000002708 enhancing effect Effects 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 239000003112 inhibitor Substances 0.000 description 4
- 238000013383 initial experiment Methods 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 230000003993 interaction Effects 0.000 description 4
- 238000011005 laboratory method Methods 0.000 description 4
- 210000002429 large intestine Anatomy 0.000 description 4
- 230000001376 precipitating effect Effects 0.000 description 4
- 230000000452 restraining effect Effects 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 241000589291 Acinetobacter Species 0.000 description 3
- 241000193830 Bacillus <bacterium> Species 0.000 description 3
- 206010041925 Staphylococcal infections Diseases 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 235000013399 edible fruits Nutrition 0.000 description 3
- 208000015688 methicillin-resistant staphylococcus aureus infectious disease Diseases 0.000 description 3
- 206010034133 Pathogen resistance Diseases 0.000 description 2
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 2
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 235000013372 meat Nutrition 0.000 description 2
- 238000005142 microbroth dilution method Methods 0.000 description 2
- 238000003908 quality control method Methods 0.000 description 2
- 238000010992 reflux Methods 0.000 description 2
- 238000012552 review Methods 0.000 description 2
- 235000014347 soups Nutrition 0.000 description 2
- 241000218202 Coptis Species 0.000 description 1
- 235000002991 Coptis groenlandica Nutrition 0.000 description 1
- 206010011409 Cross infection Diseases 0.000 description 1
- 241000588747 Klebsiella pneumoniae Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 208000037942 Methicillin-resistant Staphylococcus aureus infection Diseases 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- WKDDRNSBRWANNC-UHFFFAOYSA-N Thienamycin Natural products C1C(SCCN)=C(C(O)=O)N2C(=O)C(C(O)C)C21 WKDDRNSBRWANNC-UHFFFAOYSA-N 0.000 description 1
- 241001125929 Trisopterus luscus Species 0.000 description 1
- UGAPHEBNTGUMBB-UHFFFAOYSA-N acetic acid;ethyl acetate Chemical compound CC(O)=O.CCOC(C)=O UGAPHEBNTGUMBB-UHFFFAOYSA-N 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 230000002155 anti-virotic effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 239000002024 ethyl acetate extract Substances 0.000 description 1
- XYIBRDXRRQCHLP-UHFFFAOYSA-N ethyl acetoacetate Chemical compound CCOC(=O)CC(C)=O XYIBRDXRRQCHLP-UHFFFAOYSA-N 0.000 description 1
- 125000004494 ethyl ester group Chemical group 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 229960002182 imipenem Drugs 0.000 description 1
- ZSKVGTPCRGIANV-ZXFLCMHBSA-N imipenem Chemical compound C1C(SCC\N=C\N)=C(C(O)=O)N2C(=O)[C@H]([C@H](O)C)[C@H]21 ZSKVGTPCRGIANV-ZXFLCMHBSA-N 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229960002260 meropenem Drugs 0.000 description 1
- DMJNNHOOLUXYBV-PQTSNVLCSA-N meropenem Chemical compound C=1([C@H](C)[C@@H]2[C@H](C(N2C=1C(O)=O)=O)[C@H](O)C)S[C@@H]1CN[C@H](C(=O)N(C)C)C1 DMJNNHOOLUXYBV-PQTSNVLCSA-N 0.000 description 1
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/63—Compounds containing para-N-benzenesulfonyl-N-groups, e.g. sulfanilamide, p-nitrobenzenesulfonyl hydrazide
- A61K31/635—Compounds containing para-N-benzenesulfonyl-N-groups, e.g. sulfanilamide, p-nitrobenzenesulfonyl hydrazide having a heterocyclic ring, e.g. sulfadiazine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/7036—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin having at least one amino group directly attached to the carbocyclic ring, e.g. streptomycin, gentamycin, amikacin, validamycin, fortimicins
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention discloses Ramulus Et Folium Pithecellobii Lucidi extracts to prepare the application in anti-ESBLs-producing bacteria Escherichia coli drug.And it further discloses Ramulus Et Folium Pithecellobii Lucidi extract and is preparing the application in the anti-ESBLs-producing bacteria Escherichia coli hypersitization medicine of antibiotic.Especially as the application in amikacin and the anti-ESBLs-producing bacteria Escherichia coli hypersitization medicine of Compound New Nomin.Tests prove that Ramulus Et Folium Pithecellobii Lucidi extract and amikacin or Compound New Nomin combination are in synergistic effect.It is combined when Ramulus Et Folium Pithecellobii Lucidi extract of the present invention is less than or equal to 1/2MIC, is applied alone dosage to reduce by 75% than amikacin；It is applied alone dosage to reduce by 99.3% than Compound New Nomin.It can be used as natural antibacterials or antibiotic sensitizer, be applied to disease treatment caused by as producing ESBL Escherichia coli.New approach and alternative medicine are provided to solve the drug resistance problems of such antibiotic.
Description
Technical field
The invention belongs to field of traditional Chinese medicine pharmacy, are related to the new use of pithecellobium clypearia (Pithecellobium clypearia) extract
On the way.
Background technique
21 century is the epoch of multi-drug resistant bacteria, after antibiotic clinical application 60 years, more and more nosocomial infections and more
Weight drug-fast bacteria infection becomes a great problem of clinical antibacterial therapy at present, and Jerons in 1961 reports the 1st clinic resistance to methoxy west
Woods staphylococcus aureus (MRSA) infected patient, at present MRSA infection gradually extend over the entire globe；Chinese bacterial resistance in 2011
Property monitoring clinic in main drug-fast bacteria distribution, Escherichia coli, Klebsiella Pneumoniae ESBLs-producing bacteria (ESBL) strain
Average out to 50.7%, 38.5% respectively, the transition of drug resistance and status are concerned.In addition, being supervised according to the Ministry of Public Health, China drug resistance
2010 annual report of cooperative groups is surveyed, Acinetobacter bauamnnii and pseudomonas aeruginosa rank first in the pathogen that hospital ICU is separated,
Acinetobacter bauamnnii is up to 60.4% and 61.4% to the resistant rate of Imipenem and Meropenem respectively.In conclusion
MRSA, all kinds of bacteriums, the Acinetobacter bauamnnii of multidrug resistant and the clinical occupation rate of pseudomonas aeruginosa for producing ESBL are climbed year by year
It rises, under the pressure that resistant rate is constantly climbed to a higher point, antibiotic treatment is faced with huge challenge.For avoid bacterial resistance phenomenon into one
Step deteriorates, and experts and scholars try hard to the new inhibition bacterial growth of discovery and treatment bacterium causes the new method of disease.Existing research
Report proves that the Chinese tradition Chinese medicine such as coptis, radix scutellariae and Fructus Forsythiae etc. have certain inhibitory effect to different drug-resistant bacterias, further
What Study of Traditional Chinese Medicine inhibited drug-fast bacteria growth focuses on finding that new bacteriostasis is stronger, the wider array of Chinese medicine of drug resistance antimicrobial spectrum.
Pithecellobium clypearia (Pithecellobium clypearia Benth), scientific name bib tree are Mimosaceae pithecellobium clypearia categories
The drying sprout and leaf of plant pithecellobium clypearia, nature and flavor bitterness are cold, and it is a variety of heat toxin diseases for the treatment of that effect is clearing heat and detoxicating, hygroscopic sore
Wait unique southern medicinal material.
Having document to disclose pithecellobium clypearia and its extract at present has antivirus action.Chinese patent CN103385912A is public
The extract for having opened pithecellobium clypearia has anti-MRSA effect and antibiotic sensitization, but the patent does not refer to the Ramulus Et Folium Pithecellobii Lucidi extract
Antibacterial action to ESBLs-producing bacteria Escherichia coli (referred to as " produce ESBL Escherichia coli ") and with antibiotic associated with
Sensitization.
Summary of the invention
Present invention aims at exploitation pithecellobium clypearias to prepare the application in drug-resistance bacteria medicine.
The invention discloses Ramulus Et Folium Pithecellobii Lucidi extracts to prepare answering in anti-ESBLs-producing bacteria Escherichia coli drug
With.
And it further discloses Ramulus Et Folium Pithecellobii Lucidi extract and is preparing the anti-ESBLs-producing bacteria Escherichia coli enhanced sensitivity of antibiotic
Application in drug.Especially as amikacin and the anti-ESBLs-producing bacteria Escherichia coli enhanced sensitivity medicine of Compound New Nomin
Application in object.Present invention can apply to human medicine or medicine for animal.
Ramulus Et Folium Pithecellobii Lucidi extract of the present invention refers to pithecellobium clypearia water extract or pithecellobium clypearia ethanol extract.The extract
Can be prepared by the following method: pithecellobium clypearia coarse powder water or ethanol water extract, and resulting extracting solution is extracted with ethyl acetate again
Taking resulting extract is target product.
The beneficial effects of the present invention are: present invention firstly discloses Ramulus Et Folium Pithecellobii Lucidi extracts to resist to ESBL Escherichia coli are produced
Bacterium effect and the sensitization to such antibiotic.Tests prove that Ramulus Et Folium Pithecellobii Lucidi extract and amikacin or Compound New Nomin
Combination is in synergistic effect.It is combined when Ramulus Et Folium Pithecellobii Lucidi extract of the present invention is less than or equal to 1/2MIC, dosage is applied alone than amikacin
Reduce by 75%；It is applied alone dosage to reduce by 99.3% than Compound New Nomin.It can be used as natural antibacterials or antibiotic sensitizer, answer
For as producing disease treatment caused by ESBL Escherichia coli.New approach is provided to solve the drug resistance problems of such antibiotic
And alternative medicine.
Specific embodiment
The present invention is by following embodiments to the pharmacology of the anti-production ESBL Escherichia coli of pithecellobium clypearia water or ethanol extract
Effect is screened.
With micro-broth dilution method Ramulus Et Folium Pithecellobii Lucidi extract to produce ESBL Escherichia coli minimum inhibitory concentration (MIC) and
Minimum bactericidal concentration (MBC).
Bacterial strain: 20 plants of production ESBL Escherichia coli (ECO, number E1-E20)；Escherichia coli Quality-control strains (ECO,
ATCC25922 it) is provided by laboratory medicine portion, No.1 Hospital Affiliated to Zhongshan Univ. clinical microbiology laboratory technique room, it is attached through Zhongshan University
Belong to the detection of First Hospital clinical microbiology laboratory technique room and confirms its drug resistance.
MH broth bouillon: MH meat soup dry powder (Britain OXOID LTD.) 2.1g is settled to 100ml, NAOH adjust pH to
7.0, it is spare to set 4 DEG C of refrigerators for high pressure sterilization.
Method: the micro broth dilution method operation recommended referring to the US National clinical test standardization committee (NCCLS).
The present invention is by following methods to Ramulus Et Folium Pithecellobii Lucidi extract to amikacin, the anti-production ESBL Escherichia coli of Compound New Nomin
Sensitization investigated.
Bacterial strain: 20 plants of production ESBLs Escherichia coli (ECO, number E1-E20)；Escherichia coli Quality-control strains (ECO,
ATCC25922 it) is provided by laboratory medicine portion, No.1 Hospital Affiliated to Zhongshan Univ. clinical microbiology laboratory technique room, it is attached through Zhongshan University
Belong to the detection of First Hospital clinical microbiology laboratory technique room and confirms its drug resistance.
MH broth bouillon: MH meat soup dry powder (Britain OXOID LTD.) 2.1g is settled to 100ml, NAOH adjust pH to
7.0, it is spare to set 4 DEG C of refrigerators for high pressure sterilization.
Method: the checkerboard method operation recommended referring to the US National clinical test standardization committee (NCCLS).
The preparation method of Ramulus Et Folium Pithecellobii Lucidi extract: pithecellobium clypearia (Pithecellobium clypearia Benth) is by Guangzhou
Flower city pharmaceutical factory provides.It takes appropriate pithecellobium clypearia to break into coarse powder, with water or alcohol reflux 2 times, 2 hours every time, filters；Merge
Filtrate is concentrated to give medicinal extract (water or ethanol extract).It after taking medicinal extract to be suspended with water, is extracted with ethyl acetate, extraction three times, merges
Acetic acid ethyl acetate extract is concentrated to give acetic acid ethyl ester extract.The ethanol extract can be made by 10-95% alcohol reflux.
The technical solution that present invention will be further explained by specific examples below.
Embodiment 1
The acetic acid ethyl ester extract of pithecellobium clypearia water extract it is anti-produce ESBL Escherichia coli restraining and sterilizing bacteria test and respectively with
The united enhanced sensitivity Effect tests of amikacin, Compound New Nomin.
1. experimental method
1) measurement of minimum inhibitory concentration (MIC):
The acetic acid ethyl ester extract of pithecellobium clypearia water extract, amikacin (AMK), Compound New Nomin (SXT) are respectively in MH
A series of doubling dilutions are carried out in broth bouillon, every 50 μ l of hole, adjusting inoculation bacterium is 1.0 × 106CFU/ml, every 50 μ l bacterium of hole
Liquid.35 DEG C of cultures；24 hours, the concentration for the minimum antibacterials that no precipitating occurs was its minimum inhibitory concentration (MIC).
2) measurement of minimum bactericidal concentration (MBC):
Counting method is coated with using plate, is drawn on 50ul bacteria suspension to blood plate from the 1) hole of item asepsis growth, uniformly
Coating, 35 DEG C are cultivated 24 hours, and bacterium colony counts, minimum anti-required for so that initial experiment viable count is reduced 99.9% or more
The concentration of bacterium drug is its minimum bactericidal concentration (MBC).
By measuring the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of drug, and data count
MIC50, MIC90, MBC50, MBC90 out, to evaluate the anti-effect for producing ESBL Escherichia coli of drug.
3) checkerboard method:
Checkerboard method carries out in 96 hole sterile culture plates, by the acetic acid ethyl ester extract of pithecellobium clypearia water extract and Ah
Doubling dilution is at series of concentrations in MH broth bouillon respectively for meter Ka Xing (AMK), Compound New Nomin (SXT), respectively with two medicines
1/4MIC to 4MIC is combined respectively, and every hole A medicine B medicine adds 25 μ L, and the concentration for adjusting bacteria suspension is 1.0 × 106CFU/ml,
Every hole is inoculated with 50 μ L bacterium solutions, and 35 DEG C are incubated for for 24 hours after observation A medicine B medicine joint afterwards to the minimum inhibitory concentration for producing ESBLs Escherichia coli
(MIC)。
The calculating of FIC index:Pass through the antibacterial finger of calculating section
Number (FIC) evaluates the acetic acid ethyl ester extract of pithecellobium clypearia water extract and the interaction of Antibiotic combination antibacterial, and FIC≤
0.5 is synergistic effect, and 0.5 FIC≤1 ﹤ is summation action, and 1 FIC≤2 ﹤ are unrelated effect, and FIC ﹥ 2 is antagonism；And according to
The acetic acid ethyl ester extract of pithecellobium clypearia water extract and the best concentration ratio of antibiotic are found out in asepsis growth hole, finally evaluate monkey
The effect of the acetic acid ethyl ester extract enhancing antibiotic effect of earrings water extract.
2. experimental result
The acetic acid ethyl ester extract of pithecellobium clypearia water extract and two kinds of antibiotic (amikacin, Compound New Nomin) are to production
The extracorporeal bacteria inhibitor test of ESBL Escherichia coli the results are shown in Table 1.
The acetic acid ethyl ester extract of pithecellobium clypearia water extract is shown in Table the extracorporeal disinfecting test result for producing ESBLs Escherichia coli
2。
By statistical analysis, the acetic acid ethyl ester extract of pithecellobium clypearia water extract and two kinds of antibiotic are to production ESBL large intestine bar
The In Vitro Bacteriostasis of bacterium and MIC50, MIC90 of sterilization are shown in Table 3.
By statistical analysis, Ramulus Et Folium Pithecellobii Lucidi extract is shown in Table 4 to MBC50, MBC90 of the sterilization for producing ESBL Escherichia coli.
The FIC value and FIC of the Combination susceptibility testing of the acetic acid ethyl ester extract of pithecellobium clypearia water extract and two kinds of antibiotic
The distribution statistics of value the results are shown in Table 5, table 6.
The acetic acid ethyl ester extract of pithecellobium clypearia water extract to its MIC50 after the sensitization of two kinds of antibiotic and combination,
MIC90 is shown in Table 7-9.
Table 1
Table 2
Table 3
Table 4
Table 5
Table 6
Table 7
Table 8
Table 9
The experimental results showed that the acetic acid ethyl ester extract of pithecellobium clypearia water extract is applied alone to the MIC50 for producing ESBL Escherichia coli
It is 1600 μ g/ml, MBC50 for 1600 μ g/ml, MIC90 be 3200 μ g/ml, MBC90 is 3200 μ g/ml；
To 20 plants of production ESBL Escherichia coli, associated with the acetic acid ethyl ester extract and amikacin of pithecellobium clypearia water extract
FIC≤2 shows two medicines without antagonism, wherein there is value≤0.5 50%FIC in synergistic effect；The second of pithecellobium clypearia water extract
Acetoacetic ester extract makes amikacin MIC50 be down to 4 μ g/ml from 16 μ g/ml being applied alone when being less than or equal to 1/2MIC, reduces
75%；MIC90 reduces by 75% from being applied alone 64 μ g/ml to be down to 16 μ g/ml；
The acetic acid ethyl ester extract and Compound New Nomin of pithecellobium clypearia water extract are combined to 20 plants of production ESBL Escherichia coli FIC
≤ 2 show two medicines, two medicine without antagonism, wherein value≤0.5 25%FIC is in synergistic effect.The acetic acid of pithecellobium clypearia water extract
Ethyl ester extract makes Compound New Nomin MIC50 and MIC90 from 2432/128 μ g/ml being applied alone when being less than or equal to and MIC being applied alone
It is down to 19/1 μ g/ml, reduces 99.3%.
Embodiment 2
The acetic acid ethyl ester extract of 10% ethanol extract of pithecellobium clypearia it is anti-produce ESBL Escherichia coli restraining and sterilizing bacteria test and
Respectively with amikacin, the united enhanced sensitivity Effect tests of Compound New Nomin.
1. experimental method
1) measurement of minimum inhibitory concentration (MIC):
The acetic acid ethyl ester extract of 10% ethanol extract of pithecellobium clypearia, amikacin (AMK), Compound New Nomin (SXT) point
A series of doubling dilutions are not carried out in MH broth bouillon, every 50 μ l of hole, adjusting inoculation bacterium is 1.0 × 106CFU/ml, every hole
50 μ l bacterium solutions.35 DEG C of cultures；24 hours, the concentration for the minimum antibacterials that no precipitating occurs was its minimum inhibitory concentration (MIC).
2) measurement of minimum bactericidal concentration (MBC):
Counting method is coated with using plate, is drawn on 50ul bacteria suspension to blood plate from the 1) hole of item asepsis growth, uniformly
Coating, 35 DEG C are cultivated 24 hours, and bacterium colony counts, minimum anti-required for so that initial experiment viable count is reduced 99.9% or more
The concentration of bacterium drug is its minimum bactericidal concentration (MBC).
By measuring the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of drug, and data count
MIC50, MIC90, MBC50, MBC90 out, to evaluate the anti-effect for producing ESBL Escherichia coli of drug.
3) checkerboard method:
Checkerboard method carries out in 96 hole sterile culture plates, and the ethyl acetate of 10% ethanol extract of pithecellobium clypearia is extracted
Doubling dilution is at series of concentrations in MH broth bouillon respectively for object and amikacin (AMK), Compound New Nomin (SXT), with two
Respectively 1/4MIC to 4MIC is combined medicine respectively, and every hole A medicine B medicine adds 25 μ L, adjust bacteria suspension concentration be 1.0 ×
106CFU/ml, every hole are inoculated with 50 μ L bacterium solutions, and 35 DEG C are incubated for for 24 hours after observation A medicine B medicine joint afterwards to production ESBLs Escherichia coli most
Low Mlc (MIC).
The calculating of FIC index:Pass through the antibacterial index of calculating section
(FIC) acetic acid ethyl ester extract of 10% ethanol extract of pithecellobium clypearia and the interaction of Antibiotic combination antibacterial, FIC are evaluated
≤ 0.5 is synergistic effect, and 0.5 FIC≤1 ﹤ is summation action, and 1 FIC≤2 ﹤ are unrelated effect, and FIC ﹥ 2 is antagonism；And root
The acetic acid ethyl ester extract of 10% ethanol extract of pithecellobium clypearia and the best concentration ratio of antibiotic are found out according to asepsis growth hole, most
The effect of the acetic acid ethyl ester extract enhancing antibiotic effect of 10% ethanol extract of final review valence pithecellobium clypearia.
2. experimental result
The acetic acid ethyl ester extract of 10% ethanol extract of pithecellobium clypearia and two kinds of antibiotic (amikacin, Compound New Nomin)
10 the results are shown in Table to the extracorporeal bacteria inhibitor test for producing ESBL Escherichia coli.
The acetic acid ethyl ester extract of 10% ethanol extract of pithecellobium clypearia tests knot to the extracorporeal disinfecting for producing ESBLs Escherichia coli
Fruit is shown in Table 11.
By statistical analysis, the acetic acid ethyl ester extract of 10% ethanol extract of pithecellobium clypearia and two kinds of antibiotic are to production ESBL
The In Vitro Bacteriostasis of Escherichia coli and MIC50, MIC90 of sterilization are shown in Table 12.
By statistical analysis, Ramulus Et Folium Pithecellobii Lucidi extract is shown in Table 13 to MBC50, MBC90 of the sterilization for producing ESBL Escherichia coli.
The FIC value of the Combination susceptibility testing of the acetic acid ethyl ester extract and two kinds of antibiotic of 10% ethanol extract of pithecellobium clypearia
And the distribution statistics of FIC value the results are shown in Table 14, table 15.
The acetic acid ethyl ester extract of 10% ethanol extract of pithecellobium clypearia to after the sensitization of two kinds of antibiotic and combination its
MIC50, MIC90 are shown in Table 16-18.
Table 10
Table 11
Table 12
Table 13
Table 14
Table 15
Table 16
Table 17
Table 18
The experimental results showed that the acetic acid ethyl ester extract of 10% ethanol extract of pithecellobium clypearia is applied alone to production ESBL Escherichia coli
MIC50 be 1600 μ g/ml, it be 3200 μ g/ml, MBC90 is 3200 μ g/ml that MIC90, which is 1600 μ g/ml, MBC50,；
To 20 plants of production ESBL Escherichia coli, the acetic acid ethyl ester extract and amikacin of 10% ethanol extract of pithecellobium clypearia join
FIC≤2 shows two medicines without antagonism, wherein there is value≤0.5 50%FIC in synergistic effect；10% ethyl alcohol of pithecellobium clypearia
The acetic acid ethyl ester extract of extract makes amikacin MIC50 be down to 4 μ from 16 μ g/ml being applied alone when being less than or equal to 1/2MIC
G/ml reduces 75%；MIC90 reduces by 75% from being applied alone 64 μ g/ml to be down to 16 μ g/ml；
The acetic acid ethyl ester extract and Compound New Nomin of 10% ethanol extract of pithecellobium clypearia are combined to 20 plants of production ESBL large intestines
Bacillus FIC≤2 shows two medicines, two medicine without antagonism, and wherein value≤0.5 20%FIC is in synergistic effect.Pithecellobium clypearia water extracts
The acetic acid ethyl ester extract of object makes Compound New Nomin MIC50 and MIC90 from 2432/ be applied alone when being less than or equal to and MIC being applied alone
128 μ g/ml are down to 19/1 μ g/ml, reduce 99.3%.
Embodiment 3
The acetic acid ethyl ester extract of 60% ethanol extract of pithecellobium clypearia it is anti-produce ESBL Escherichia coli restraining and sterilizing bacteria test and
Respectively with amikacin, the united enhanced sensitivity Effect tests of Compound New Nomin.
1. experimental method
1) measurement of minimum inhibitory concentration (MIC):
The acetic acid ethyl ester extract of 60% ethanol extract of pithecellobium clypearia, amikacin (AMK), Compound New Nomin (SXT) point
A series of doubling dilutions are not carried out in MH broth bouillon, every 50 μ l of hole, adjusting inoculation bacterium is 1.0 × 106CFU/ml, every hole
50 μ l bacterium solutions.35 DEG C of cultures；24 hours, the concentration for the minimum antibacterials that no precipitating occurs was its minimum inhibitory concentration (MIC).
2) measurement of minimum bactericidal concentration (MBC):
Counting method is coated with using plate, is drawn on 50ul bacteria suspension to blood plate from the 1) hole of item asepsis growth, uniformly
Coating, 35 DEG C are cultivated 24 hours, and bacterium colony counts, minimum anti-required for so that initial experiment viable count is reduced 99.9% or more
The concentration of bacterium drug is its minimum bactericidal concentration (MBC).
By measuring the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of drug, and data count
MIC50, MIC90, MBC50, MBC90 out, to evaluate the anti-effect for producing ESBL Escherichia coli of drug.
3) checkerboard method:
Checkerboard method carries out in 96 hole sterile culture plates, and the ethyl acetate of 60% ethanol extract of pithecellobium clypearia is extracted
Doubling dilution is at series of concentrations in MH broth bouillon respectively for object and amikacin (AMK), Compound New Nomin (SXT), with two
Respectively 1/4MIC to 4MIC is combined medicine respectively, and every hole A medicine B medicine adds 25 μ L, adjust bacteria suspension concentration be 1.0 ×
106CFU/ml, every hole are inoculated with 50 μ L bacterium solutions, and 35 DEG C are incubated for for 24 hours after observation A medicine B medicine joint afterwards to production ESBLs Escherichia coli most
Low Mlc (MIC).
The calculating of FIC index:Pass through the antibacterial index of calculating section
(FIC) acetic acid ethyl ester extract of 60% ethanol extract of pithecellobium clypearia and the interaction of Antibiotic combination antibacterial, FIC are evaluated
≤ 0.5 is synergistic effect, and 0.5 FIC≤1 ﹤ is summation action, and 1 FIC≤2 ﹤ are unrelated effect, and FIC ﹥ 2 is antagonism；And root
The acetic acid ethyl ester extract of 60% ethanol extract of pithecellobium clypearia and the best concentration ratio of antibiotic are found out according to asepsis growth hole, most
The effect of the acetic acid ethyl ester extract enhancing antibiotic effect of 60% ethanol extract of final review valence pithecellobium clypearia.
2. experimental result
The acetic acid ethyl ester extract of 60% ethanol extract of pithecellobium clypearia and two kinds of antibiotic (amikacin, Compound New Nomin)
19 the results are shown in Table to the extracorporeal bacteria inhibitor test for producing ESBL Escherichia coli.
The acetic acid ethyl ester extract of 60% ethanol extract of pithecellobium clypearia tests knot to the extracorporeal disinfecting for producing ESBLs Escherichia coli
Fruit is shown in Table 20.
By statistical analysis, the acetic acid ethyl ester extract of 60% ethanol extract of pithecellobium clypearia and two kinds of antibiotic are to production ESBL
The In Vitro Bacteriostasis of Escherichia coli and MIC50, MIC90 of sterilization are shown in Table 21.
By statistical analysis, Ramulus Et Folium Pithecellobii Lucidi extract is shown in Table 22 to MBC50, MBC90 of the sterilization for producing ESBL Escherichia coli.
The FIC value of the Combination susceptibility testing of the acetic acid ethyl ester extract and two kinds of antibiotic of 60% ethanol extract of pithecellobium clypearia
And the distribution statistics of FIC value the results are shown in Table 23, table 24.
The acetic acid ethyl ester extract of 60% ethanol extract of pithecellobium clypearia to after the sensitization of two kinds of antibiotic and combination its
MIC50, MIC90 are shown in Table 25-27.
Table 19
Table 20
Table 21
Table 22
Table 23
Table 24
Table 25
Table 26
Table 27
The experimental results showed that the acetic acid ethyl ester extract of 60% ethanol extract of pithecellobium clypearia is applied alone to production ESBL Escherichia coli
MIC50 be 800 μ g/ml, it be 1600 μ g/ml, MBC90 is 1600 μ g/ml that MIC90, which is 800 μ g/ml, MBC50,；
To 20 plants of production ESBL Escherichia coli, the acetic acid ethyl ester extract and amikacin of 60% ethanol extract of pithecellobium clypearia join
FIC≤2 shows two medicines without antagonism, wherein there is value≤0.5 70%FIC in synergistic effect；60% ethyl alcohol of pithecellobium clypearia
The acetic acid ethyl ester extract of extract makes amikacin MIC50 be down to 4 μ from 16 μ g/ml being applied alone when being less than or equal to 1/4MIC
G/ml reduces 75%；MIC90 is down to 8 μ g/ml from 64 μ g/ml are applied alone；
The acetic acid ethyl ester extract and Compound New Nomin of 60% ethanol extract of pithecellobium clypearia are combined to 20 plants of production ESBL large intestines
Bacillus FIC≤2 shows two medicines, two medicine without antagonism, and wherein value≤0.5 30%FIC is in synergistic effect.60% second of pithecellobium clypearia
The acetic acid ethyl ester extract of alcohol extracting thing makes Compound New Nomin MIC50 and MIC90 from being applied alone when being less than or equal to and MIC being applied alone
2432/128 μ g/ml is down to 19/1 μ g/ml, reduces 99.3%.
Embodiment 4
The acetic acid ethyl ester extract of 95% ethanol extract of pithecellobium clypearia it is anti-produce ESBL Escherichia coli restraining and sterilizing bacteria test and
Respectively with amikacin, the united enhanced sensitivity Effect tests of Compound New Nomin.
1. experimental method
1) measurement of minimum inhibitory concentration (MIC):
The acetic acid ethyl ester extract of 95% ethanol extract of pithecellobium clypearia, amikacin (AMK), Compound New Nomin (SXT) point
A series of doubling dilutions are not carried out in MH broth bouillon, every 50 μ l of hole, adjusting inoculation bacterium is 1.0 × 106CFU/ml, every hole
50 μ l bacterium solutions.35 DEG C of cultures；24 hours, the concentration for the minimum antibacterials that no precipitating occurs was its minimum inhibitory concentration (MIC).
2) measurement of minimum bactericidal concentration (MBC):
Counting method is coated with using plate, is drawn on 50ul bacteria suspension to blood plate from the 1) hole of item asepsis growth, uniformly
Coating, 35 DEG C are cultivated 24 hours, and bacterium colony counts, minimum anti-required for so that initial experiment viable count is reduced 99.9% or more
The concentration of bacterium drug is its minimum bactericidal concentration (MBC).
By measuring the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of drug, and data count
MIC50, MIC90, MBC50, MBC90 out, to evaluate the anti-effect for producing ESBL Escherichia coli of drug.
3) checkerboard method:
Checkerboard method carries out in 96 hole sterile culture plates, and the ethyl acetate of 95% ethanol extract of pithecellobium clypearia is extracted
Doubling dilution is at series of concentrations in MH broth bouillon respectively for object and amikacin (AMK), Compound New Nomin (SXT), with two
Respectively 1/4MIC to 4MIC is combined medicine respectively, and every hole A medicine B medicine adds 25 μ L, adjust bacteria suspension concentration be 1.0 ×
106CFU/ml, every hole are inoculated with 50 μ L bacterium solutions, and 35 DEG C are incubated for for 24 hours after observation A medicine B medicine joint afterwards to production ESBLs Escherichia coli most
Low Mlc (MIC).
The calculating of FIC index:Pass through the antibacterial finger of calculating section
Number (FIC) evaluates the acetic acid ethyl ester extract of 95% ethanol extract of pithecellobium clypearia and the interaction of Antibiotic combination antibacterial,
FIC≤0.5 is synergistic effect, and 0.5 FIC≤1 ﹤ is summation action, and 1 FIC≤2 ﹤ are unrelated effect, and FIC ﹥ 2 is antagonism；And
The acetic acid ethyl ester extract of 95% ethanol extract of pithecellobium clypearia and the best concentration ratio of antibiotic are found out according to asepsis growth hole,
The effect of the acetic acid ethyl ester extract enhancing antibiotic effect of final evaluation 95% ethanol extract of pithecellobium clypearia.
2. experimental result
The acetic acid ethyl ester extract of 95% ethanol extract of pithecellobium clypearia and two kinds of antibiotic (amikacin, Compound New Nomin)
28 the results are shown in Table to the extracorporeal bacteria inhibitor test for producing ESBL Escherichia coli.
The acetic acid ethyl ester extract of 95% ethanol extract of pithecellobium clypearia tests knot to the extracorporeal disinfecting for producing ESBLs Escherichia coli
Fruit is shown in Table 29.
By statistical analysis, the acetic acid ethyl ester extract of 95% ethanol extract of pithecellobium clypearia and two kinds of antibiotic are to production ESBL
The In Vitro Bacteriostasis of Escherichia coli and MIC50, MIC90 of sterilization are shown in Table 30.
By statistical analysis, Ramulus Et Folium Pithecellobii Lucidi extract is shown in Table 31 to MBC50, MBC90 of the sterilization for producing ESBL Escherichia coli.
The FIC value of the Combination susceptibility testing of the acetic acid ethyl ester extract and two kinds of antibiotic of 95% ethanol extract of pithecellobium clypearia
And the distribution statistics of FIC value the results are shown in Table 32, table 33.
The acetic acid ethyl ester extract of 95% ethanol extract of pithecellobium clypearia to after the sensitization of two kinds of antibiotic and combination its
MIC50, MIC90 are shown in Table 34-36.
Table 28
Table 29
Table 30
Table 31
Table 32
Table 33
Table 34
Table 35
Table 36
The experimental results showed that the acetic acid ethyl ester extract of 95% ethanol extract of pithecellobium clypearia is applied alone to production ESBL Escherichia coli
MIC50 be 1600 μ g/ml, it be 3200 μ g/ml, MBC90 is 3200 μ g/ml that MIC90, which is 1600 μ g/ml, MBC50,；
To 20 plants of production ESBL Escherichia coli, the acetic acid ethyl ester extract and amikacin of 95% ethanol extract of pithecellobium clypearia join
FIC≤2 shows two medicines without antagonism, wherein there is value≤0.5 50%FIC in synergistic effect；95% ethyl alcohol of pithecellobium clypearia
The acetic acid ethyl ester extract of extract makes amikacin MIC50 be down to 4 μ from 16 μ g/ml being applied alone when being less than or equal to 1/2MIC
G/ml reduces 75%；MIC90 reduces by 75% from being applied alone 64 μ g/ml to be down to 16 μ g/ml；
The acetic acid ethyl ester extract and Compound New Nomin of 95% ethanol extract of pithecellobium clypearia are combined to 20 plants of production ESBL large intestines
Bacillus FIC≤2 shows two medicines, two medicine without antagonism, and wherein value≤0.5 20%FIC is in synergistic effect.95% second of pithecellobium clypearia
The acetic acid ethyl ester extract of alcohol extracting thing makes Compound New Nomin MIC50 and MIC90 from being applied alone when being less than or equal to and MIC being applied alone
2432/128 μ g/ml is down to 19/1 μ g/ml, reduces 99.3%.
Claims (4)
1. Ramulus Et Folium Pithecellobii Lucidi extract is preparing the application in the anti-ESBLs-producing bacteria Escherichia coli hypersitization medicine of antibiotic, institute
The antibiotic stated is amikacin or Compound New Nomin.
2. application as described in claim 1, which is characterized in that the Ramulus Et Folium Pithecellobii Lucidi extract is prepared by following methods: monkey ear
Ring coarse powder water or ethanol water extract, and resulting extracting solution is extracted with ethyl acetate again, and resulting extract is target
Product.
3. application as claimed in claim 2, which is characterized in that the ethanol water is to count by volume, and concentration is
The ethanol water of 10%-95%.
4. application as claimed in claim 3, which is characterized in that the ethanol water is to count by volume, and concentration is
60% ethanol water.
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| CN201610308718.6A CN105998153B (en) | 2016-05-10 | 2016-05-10 | Ramulus Et Folium Pithecellobii Lucidi extract is preparing the application in anti-ESBLs-producing bacteria Escherichia coli drug |
| PCT/CN2017/071671 WO2017193635A1 (en) | 2016-05-10 | 2017-01-19 | Pithecellobium clypearia benth. extract and application for preparing anti-microbial agent |
| EP17795261.1A EP3456335B1 (en) | 2016-05-10 | 2017-01-19 | Pithecellobium clypearia benth extract and application for preparing anti-microbial agent |
| US15/920,480 US11154582B2 (en) | 2016-05-10 | 2018-03-14 | Method of chinese herbal medicine extract used for treating multiple diseases caused by drug resistant bacteria infection |
| US17/129,885 US20210106640A1 (en) | 2016-05-10 | 2020-12-21 | Method of chinese herbal medicine extract used for treating multiple diseases caused by drug resistant bacteria infection |
| US17/129,901 US20210106641A1 (en) | 2016-05-10 | 2020-12-21 | Method of chinese herbal medicine extract used for treating multiple diseases caused by drug resistant bacteria infection |
| US17/129,853 US11491198B2 (en) | 2016-05-10 | 2020-12-21 | Method of chinese herbal medicine extract used for treating multiple diseases caused by drug resistant bacteria infection |
| US17/494,850 US11654174B2 (en) | 2016-05-10 | 2021-10-06 | Method of chinese herbal medicine extract used for treating multiple diseases caused by drug resistant bacteria infection |
| US17/494,858 US11793849B2 (en) | 2016-05-10 | 2021-10-06 | Method of Chinese herbal medicine extract used for treating multiple diseases caused by drug resistant bacteria infection |
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| CN1565455A (en) * | 2003-07-04 | 2005-01-19 | 成都博瑞医药科技开发有限公司 | Antibiotics compositions for curing broad spectrum beta-lactamase bacteria infection |
| CN1795888A (en) * | 2004-12-29 | 2006-07-05 | 广州莱泰制药有限公司 | Application of Extractive of Abarema elliptica in use for preparing antiallergic medication, food and cosmetic |
| CN101032547A (en) * | 2006-03-07 | 2007-09-12 | 广东奇方药业有限公司 | Anti-inflammatory and antivirotic medicine composition |
| CN103385912A (en) * | 2013-07-24 | 2013-11-13 | 中山大学 | Pithecellobium clypearia extracts and application of extract in preparation of medicines for treating methicillin-resistant staphylococcus aureus |
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| CN1795888A (en) * | 2004-12-29 | 2006-07-05 | 广州莱泰制药有限公司 | Application of Extractive of Abarema elliptica in use for preparing antiallergic medication, food and cosmetic |
| CN101032547A (en) * | 2006-03-07 | 2007-09-12 | 广东奇方药业有限公司 | Anti-inflammatory and antivirotic medicine composition |
| CN103385912A (en) * | 2013-07-24 | 2013-11-13 | 中山大学 | Pithecellobium clypearia extracts and application of extract in preparation of medicines for treating methicillin-resistant staphylococcus aureus |
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