WO2017193635A1 - 猴耳环提取物在制备抗菌药物中的应用 - Google Patents

猴耳环提取物在制备抗菌药物中的应用 Download PDF

Info

Publication number
WO2017193635A1
WO2017193635A1 PCT/CN2017/071671 CN2017071671W WO2017193635A1 WO 2017193635 A1 WO2017193635 A1 WO 2017193635A1 CN 2017071671 W CN2017071671 W CN 2017071671W WO 2017193635 A1 WO2017193635 A1 WO 2017193635A1
Authority
WO
WIPO (PCT)
Prior art keywords
monkey
mic
earrings
extract
ethanol extract
Prior art date
Application number
PCT/CN2017/071671
Other languages
English (en)
French (fr)
Other versions
WO2017193635A9 (zh
Inventor
苏薇薇
刘翀
周倩
李沛波
彭维
王永刚
Original Assignee
中山大学
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from CN201610308207.4A external-priority patent/CN105920082B/zh
Priority claimed from CN201610308717.1A external-priority patent/CN105816511B/zh
Priority claimed from CN201610308718.6A external-priority patent/CN105998153B/zh
Priority claimed from CN201610308716.7A external-priority patent/CN105963339B/zh
Priority to EP17795261.1A priority Critical patent/EP3456335B1/en
Application filed by 中山大学 filed Critical 中山大学
Publication of WO2017193635A1 publication Critical patent/WO2017193635A1/zh
Publication of WO2017193635A9 publication Critical patent/WO2017193635A9/zh
Priority to US15/920,480 priority patent/US11154582B2/en
Priority to US17/129,885 priority patent/US20210106640A1/en
Priority to US17/129,853 priority patent/US11491198B2/en
Priority to US17/129,901 priority patent/US20210106641A1/en
Priority to US17/494,850 priority patent/US11654174B2/en
Priority to US17/494,858 priority patent/US11793849B2/en

Links

Classifications

    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • 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

Definitions

  • the invention relates to a new use of monkey earring extract, in particular to the application of monkey earring extract in preparing various antibiotic resistant drugs.
  • Acinetobacter baumannii and Pseudomonas aeruginosa are the first among the pathogens isolated from hospital ICU, and the resistance rate of Acinetobacter baumannii to imipenem and meropenem Up to 60.4% and 61.4% respectively.
  • the clinical market share of MRSA, ESBL-producing bacteria, multi-drug resistant Acinetobacter baumannii and Pseudomonas aeruginosa has increased year by year, and antibiotic treatment is facing pressure under the pressure of increasing resistance. huge challenge.
  • the present invention discloses the application of the monkey earring extract in preparing anti-resistant bacteria and its sensitizing effect in combination with similar antibiotics, specifically:
  • monkey earring extract in the preparation of anti-multidrug-resistant Acinetobacter baumannii drugs or multi-drug resistant Acinetobacter baumannii antibiotic sensitizing drugs.
  • Monkey earring extract combined with antibiotic imipenem or tetracycline or polymyxin B or ceftazidime or levofloxacin showed significant sensitization.
  • monkey earring extract in the preparation of anti-multidrug-resistant Pseudomonas aeruginosa drugs or anti-multidrug-resistant Pseudomonas aeruginosa antibiotic sensitizing drugs.
  • Monkey earring extract combined with antibiotic levofloxacin or imipenem or amikacin or ceftazidime or cefoperazone showed significant sensitization. .
  • monkey earring extract in the preparation of an antibiotic-producing antibiotic-producing antibiotic-promoting drug for the production of extended-spectrum ⁇ -lactamase-producing Escherichia coli or anti-producing extended-spectrum ⁇ -lactamase.
  • Monkey earring extract combined with antibiotic amikacin or compound sulfamethoxazole showed significant sensitization.
  • the monkey earring extract in the preparation of a drug resistant to extended-spectrum ⁇ -lactamase Klebsiella pneumoniae.
  • the monkey earring extract has obvious antibacterial effect on the production of extended-spectrum ⁇ -lactamase Klebsiella pneumoniae.
  • the monkey earring extract is preferably a monkey earring water extract or a monkey earring ethanol extract.
  • the monkey earring extract is prepared by using a monkey earring coarse powder with water or by volume.
  • the aqueous extract was extracted with 10% to 95% of ethanol, and the obtained extract was extracted with ethyl acetate, and the obtained extract was the target product.
  • the aqueous ethanol solution is preferably an aqueous ethanol solution having a concentration of 60% by volume.
  • the drugs referred to in the present invention include human or animal drugs, or soil treatment drugs or related preparations.
  • the present invention discloses for the first time that the monkey earring extract is resistant to multi-drug resistant Acinetobacter baumannii, multi-drug resistant Pseudomonas aeruginosa, extended-spectrum ⁇ -lactamase-producing Escherichia coli, and extended-spectrum ⁇ -lactamase producing pneumonia.
  • the combination of monkey earring extract and levofloxacin or imipenem or amikacin or ceftazidime or cefoperazone has a synergistic effect against multidrug-resistant Pseudomonas aeruginosa, reducing the use of antibiotics by 50% to 99.2%. .
  • the combination of monkey earring extract and amikacin or compound sulfamethoxazole has synergistic effect against E. coli produced by extended-spectrum ⁇ -lactamase, which reduces the amount of antibiotics by 75% to 99.3%.
  • the monkey earring extract of the present invention can be used as a sensitizer for a natural antibacterial agent or a similar antibiotic of the above-mentioned resistant bacteria, and is applied to the treatment of diseases caused by the above bacteria.
  • the present invention provides new ways and alternative drugs for solving the problem of drug resistance of such antibiotics. It is suitable for human medicine, other animal medicines and soil treatment for corresponding infectious bacteria. It is a natural plant extract with no side effects and the extraction method is simple and environmentally friendly.
  • the invention further clarifies the pharmacological action of the monkey earring extract anti-resistant bacteria and the sensitizing effect on the same antibiotic by the following embodiments.
  • EA Monkey Earring Extract
  • the monkey earring (Pithecellobium clypearia Benth) was supplied by Guangzhou Huacheng Pharmaceutical Factory. Take appropriate amount of monkey earrings into a coarse powder, reflux twice with water or 10%-95% aqueous ethanol solution for 2 hours each time, filter; combine the filtrate and concentrate to obtain extract (ie monkey earring water or ethanol extract).
  • the extract was suspended with water, extracted with ethyl acetate and extracted three times.
  • the extract obtained by refluxing with a 10% aqueous solution of ethanol is hereinafter referred to as a 10% ethanol extract of monkey earrings, and the extract obtained by refluxing other concentrations of aqueous ethanol solution is so analogous.
  • test strains of the present invention were all provided by the clinical microbiology laboratory of the First affiliated Hospital of Sun Yat-sen University, and the drug resistance was confirmed by the clinical microbiology laboratory of the First affiliated Hospital of Sun Yat-sen University.
  • MH broth medium 2.1 g of MH broth dry powder (OXOID LTD., UK), the volume was adjusted to 100 ml, the pH of NAOH was adjusted to 7.0, autoclaved, and placed in a refrigerator at 4 °C for use.
  • MIC minimum inhibitory concentration
  • MMC minimum bactericidal concentration
  • NCLS National Committee for Clinical Trial Standardization
  • Monkey earring extract, imipenem (IMP), tetracycline (TE), polymyxin B (POLB), ceftazidime (CAZ), levofloxacin (LVX) were separately diluted in MH broth medium. 50 ⁇ l per well, the inoculum was adjusted to 1.0 ⁇ 10 6 CFU/ml, and 50 ⁇ l of the bacterial solution was added to each well. Culture at 35 ° C; 24 hours, the concentration of the lowest antibacterial drug without precipitation is its minimum inhibitory concentration (MIC).
  • MIC minimum inhibitory concentration
  • the concentration of the minimum antimicrobial agent required or above is its minimum bactericidal concentration (MBC).
  • the anti-multidrug-resistant Acinetobacter baumannii was evaluated by determining the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the drug and counting the data to obtain MIC 50 , MIC 90 , MBC 50 , and MBC 90 .
  • MIC minimum inhibitory concentration
  • MBC minimum bactericidal concentration
  • the checkerboard dilution method was carried out in a 96-well sterile culture plate.
  • the monkey earring extract and imipenem (IMP), tetracycline (TE), polymyxin B (POLB), and ceftazidime (CAZ) were respectively in MH broth.
  • the medium was diluted to a series concentration, and each of the two drugs was combined with 1/4 MIC to 4 MIC, and 25 ⁇ L of each drug A was added, and the concentration of the bacterial suspension was adjusted to 1.0 ⁇ 10 6 CFU/ml.
  • 50 ⁇ L of bacterial solution was inoculated and incubated at 35 ° C for 24 h to observe the minimum inhibitory concentration (MIC) of multidrug-resistant Acinetobacter baumannii after combination of drug A and drug B.
  • MIC minimum inhibitory concentration
  • FIC index The interaction between the monkey earring extract and the antibiotic combined antibacterial was evaluated by calculating the partial inhibition index (FIC).
  • FIC ⁇ 0.5 is synergistic, 0.5 ⁇ FIC ⁇ 1 is additive, 1 ⁇ FIC ⁇ 2 is irrelevant, FIC >2 is antagonistic; and the optimal concentration ratio of monkey earring extract and antibiotic is found according to the sterile growth hole, and finally the effect of monkey earring extract on enhancing the antibiotic efficacy is evaluated.
  • test articles of the present example include: monkey earring water extract, monkey earrings 10% ethanol extract, monkey earrings 60% ethanol extract, monkey earrings 95% ethanol extract.
  • Multidrug-resistant Acinetobacter baumannii (MDRAB), numbered A1-A20.
  • the MIC 50 and MIC 90 statistical analysis of the in vitro bacteriostasis and bactericidal action of monkey earring water extract and five antibiotics against multidrug-resistant Acinetobacter baumannii are shown in Table A3.
  • the FIC of the monkey earring water extract combined with imipenem ⁇ 1 indicates that the two drugs have a synergistic effect or a partial synergistic effect, wherein 35% FIC value ⁇ 0.5 is synergistic; 20 multi-drug resistant Bauman does not move Bacillus, monkey earring water extract reduced the imipenem MIC 50 from 32 ⁇ g/ml for single use to 8 ⁇ g/ml at a concentration of ⁇ MIC alone; the MIC 90 decreased from 32 ⁇ g/ml to 16 ⁇ g/ Ml, reduced by 50%;
  • the combination of monkey earring water extract and tetracycline on 20 multi-drug resistant Acinetobacter baumannii FIC ⁇ 1 indicates that the two drugs have synergistic effect or partial synergistic effect, 35% FIC value ⁇ 0.5 synergistic effect;
  • the drug-resistant Acinetobacter baumannii, monkey earring water extract reduced the tetracycline MIC 50 to 256 ⁇ g/ml to 64 ⁇ g/ml, which was reduced by 75%, and the MIC 90 decreased from 512 ⁇ g/ml to 128 ⁇ g/ Ml, reduced by 75%;
  • the combination of water extract of monkey earrings and polymyxin B with FIC ⁇ 2 indicates that the two drugs have no antagonistic effect, 25% of which have a synergistic effect of FIC ⁇ 0.5;
  • the drug-resistant Acinetobacter baumannii, monkey earring water extract reduced the polymyxin BMIC 50 to 4 ⁇ g/ml by 4 ⁇ g/ml, reduced by 75%, and the MIC 90 decreased from 16 ⁇ g/ml. Up to 7 ⁇ g/ml, reduced by 75%;
  • monkey earring water extract can be ⁇ when used alone MIC, ceftazidime MIC 50 so reduced 128 ⁇ g / ml from 512 ⁇ g / ml, decreased by 75%; MIC 90 reduced to 256 ⁇ g / ml from 512 ⁇ g / ml, a 50% decrease;
  • the monkey earring water extract combined with levofloxacin against 20 strains of multi-drug resistant Acinetobacter baumannii FIC ⁇ 2 indicates that the two drugs have no antagonistic effect, and 35% of the FIC value ⁇ 0.5 indicates that the two drugs have a certain degree of synergistic effect;
  • the monkey earring water extract reduced the levofloxacin MIC 50 to 8 ⁇ g/ml to 2 ⁇ g/ml, a reduction of 75%, and the MIC 90 from 32 ⁇ g/ml. Dropped to 8 ⁇ g/ml, reduced by 75%.
  • the MIC 50 and MIC 90 statistical analysis of the in vitro bacteriostasis and bactericidal activity of the monkey earrings 10% ethanol extract and five antibiotics against the multi-drug resistant Acinetobacter baumannii are shown in Table A15.
  • the monkey earring extract against the multi-drug resistant Acinetobacter baumannii bactericidal MBC 50 , MBC 90 statistical analysis is shown in Table A16.
  • the FIC of the 10% ethanol extract of monkey earrings combined with imipenem ⁇ 1 indicates that the two drugs have synergistic effect or partial synergistic effect, 45% of which have a synergistic effect of FIC value ⁇ 0.5; 20 pairs of multi-drug resistant Bauman Acinetobacter, monkey earrings 10% ethanol extract at a concentration of ⁇ 1 / 2 the MIC alone, imipenem MIC 50 so reduced 8 ⁇ g / ml from a single use of 32 ⁇ g / ml, decreased 75%; MIC 90 from 32 ⁇ g/ml reduced to 16 ⁇ g/ml, reduced by 50%;
  • the combination of 10% ethanol extract of monkey earrings and ceftazidime on 20 multi-drug resistant Acinetobacter baumannii FIC ⁇ 2 indicates that there is no antagonistic effect of the two drugs, 30% FIC ⁇ 0.5 synergistic effect; monkey earrings 10% ethanol extraction
  • the ethyl acetate extract of the product can reduce the ceftazidime MIC 50 from 512 ⁇ g/ml to 128 ⁇ g/ml, which is reduced by 75% when ⁇ 1/2 of the MIC alone; the MIC 90 decreases from 512 ⁇ g/ml to 256 ⁇ g/ml. 50%;
  • the combination of 10% ethanol extract of monkey earrings and levofloxacin combined with anti-20 strains of multi-drug resistant Acinetobacter baumannii FIC ⁇ 2 indicates that the two drugs have no antagonistic effect, and 20% FIC value ⁇ 0.5 indicates that the two drugs have a certain degree of synergy.
  • the MIC 50 and MIC 90 statistical analysis of the in vitro antibacterial and bactericidal effects of monkey earrings 60% ethanol extract and five antibiotics against multidrug-resistant Acinetobacter baumannii are shown in Table A27.
  • the FIC of the 60% ethanol extract of monkey earrings combined with imipenem ⁇ 1 indicates that the two drugs have a synergistic effect or a partial synergistic effect, of which 70% FIC value ⁇ 0.5 is synergistic; 20 multi-drug resistant Bauman Acinetobacter, monkey earrings 60% ethanol extract at a concentration of ⁇ 1 / 2 the MIC alone, imipenem MIC 50 so reduced 8 ⁇ g / ml from a single use of 32 ⁇ g / ml, decreased 75%; MIC 90 from 32 ⁇ g/ml reduced to 16 ⁇ g/ml, reduced by 50%;
  • monkey earrings 60% ethanol extraction composition may be at ⁇ 1 / 2 alone MIC, ceftazidime MIC 50 so reduced 128 ⁇ g / ml from 512 ⁇ g / ml, decreased by 75%; MIC 90 reduced to 256 ⁇ g / ml from 512 ⁇ g / ml, a 50% decrease;
  • Monkey earrings 60% ethanol extract combined with levofloxacin anti-20 strains of multi-drug resistant Acinetobacter baumannii FIC ⁇ 2 indicates that the two drugs have no antagonistic effect, of which 5% FIC value ⁇ 0.5 indicates that the two drugs have a certain degree of synergy Role; for 20 multi-drug resistant Acinetobacter baumannii, 60% ethanol extract of monkey earrings reduced the levofloxacin MIC 50 to 8 ⁇ g/ml by 4 ⁇ g/ml, less than or equal to MIC alone, by 50%, MIC 90 was reduced from 32 ⁇ g/ml to 8 ⁇ g/ml, a 75% reduction.
  • the MIC 50 and MIC 90 statistical analysis of the in vitro antibacterial and bactericidal effects of monkey earrings 95% ethanol extract and five antibiotics against multidrug-resistant Acinetobacter baumannii are shown in Table A39.
  • the monkey earring extract against the multi-drug resistant Acinetobacter baumannii bactericidal MBC 50 , MBC 90 statistical analysis is shown in Table A40.
  • the FIC of the 95% ethanol extract of monkey earrings combined with imipenem ⁇ 1 indicates that the two drugs have synergistic or partial synergistic effects, of which 55% FIC value ⁇ 0.5 is synergistic; 20 multi-drug resistant Bauman Acinetobacter, monkey earrings 95% ethanol extract reduced imipenem MIC 50 from 32 ⁇ g/ml for single use to 8 ⁇ g/ml at a concentration of ⁇ MIC alone; 75% reduction; MIC 90 from 32 ⁇ g/ml Reduced to 16 ⁇ g/ml, reduced by 50%;
  • the monkey earring water extract alone has an MIC 50 of 800 ⁇ g/ml for multidrug-resistant Pseudomonas aeruginosa, 1600 ⁇ g/ml for MIC 90, 1600 ⁇ g/ml for MBC 50, and 1600 ⁇ g/ml for MBC 90 ;
  • monkey earring water extract reduced the cefoperazone MIC 50 from 64 ⁇ g/ml for single use to 0.5 ⁇ g/ml at a concentration of ⁇ MIC alone, which was reduced by 99.2%; MIC 90 from 512 ⁇ g/ml to 256 ⁇ g/ml, a 50% reduction;
  • the combination of monkey earring water extract and amikacin FIC ⁇ 2 indicates that the two drugs have no antagonistic effect, 35% of FIC ⁇ 0.5 synergistic effect; 20 strains of multi-drug resistant multi-drug resistant Pseudomonas aeruginosa, monkey earrings
  • the water extract reduced the amikacin MIC 50 from 1 ⁇ g/ml for single use to 0.0625 ⁇ g/ml, which was reduced by 93.75%, and the MIC 90 decreased from 32 ⁇ g/ml to 2 ⁇ g/ml. 93.75%;
  • the FIC of the monkey earring water extract combined with imipenem ⁇ 2 indicates that the two drugs have no antagonistic effect, 40% of which has a synergistic effect of FIC ⁇ 0.5; 20 strains of multi-drug resistant multi-drug resistant Pseudomonas aeruginosa, monkey
  • the water extract of earrings reduced the imipenem MIC 50 from 32 ⁇ g/ml for single use to 2 ⁇ g/ml, which was reduced by 93.75%, and the MIC 90 decreased from 64 ⁇ g/ml to 16 ⁇ g/ml. 75%;
  • 20 multi-drug resistant multidrug-resistant Pseudomonas aeruginosa the water extract of monkey earrings reduced the levofloxacin MIC 50 from 4 ⁇ g/ml to 1 ⁇ g/ml, a reduction of 75% at a concentration of ⁇ MIC alone; MIC 90 was reduced from 32 ⁇ g/ml to 8 ⁇ g/ml, which was reduced by 75%.
  • the MIC 50 and MIC 90 of the antibacterial and bactericidal activity of the multi-drug resistant Pseudomonas aeruginosa 10% ethanol extract and five antibiotics in monkey earrings are shown in Table P15.
  • the MBC 50 and MBC 90 of the monkey earring extract for sterilizing multi-drug resistant P. aeruginosa are shown in Table P16.
  • the monkey earring 10% ethanol extract alone has an MIC 50 of 800 ⁇ g/ml for multidrug-resistant Pseudomonas aeruginosa, 1600 ⁇ g/ml for MIC 90, 1600 ⁇ g/ml for MBC 50, and 1600 ⁇ g/ml for MBC 90 ;
  • Multi-drug resistant multi-drug resistant P. aeruginosa monkey earrings 10% ethanol extract at a concentration of ⁇ MIC alone, reducing the ceftazidime MIC 50 from 32 ⁇ g/ml for single use to 16 ⁇ g/ml, a 50% reduction; MIC 90 decreased from 256 ⁇ g/ml to 64 ⁇ g/ml, a 75% reduction;
  • the combination of 10% ethanol extract of monkey earrings and imipenem after FIC ⁇ 2 indicates that the two drugs have no antagonistic effect, 50% of FIC ⁇ 0.5 synergistic effect; 20 strains of multi-drug resistant multi-drug resistant Pseudomonas aeruginosa
  • the monkey earrings 10% ethanol extract reduced the imipenem MIC 50 from 32 ⁇ g/ml for single use to 2 ⁇ g/ml at a concentration of ⁇ MIC, which was reduced by 93.75%; the MIC 90 decreased from 64 ⁇ g/ml to 16 ⁇ g. /ml, reduced by 75%;
  • the MIC 50 and MIC 90 statistical analysis of the in vitro bacteriostasis and bactericidal activity of the monkey earrings 60% ethanol extract and five antibiotics against multidrug-resistant Pseudomonas aeruginosa are shown in Table P27.
  • the MBC 50 and MBC 90 statistical analysis of the sterilization of the monkey earring extract against the multidrug-resistant Pseudomonas aeruginosa are shown in Table P28.
  • Monkey earrings 60% ethanol extract alone has a MIC 50 of 400 ⁇ g/ml for multidrug-resistant Pseudomonas aeruginosa, 800 ⁇ g/ml for MIC 90, 1600 ⁇ g/ml for MBC 50 and 1600 ⁇ g/ml for MBC 90 ;
  • Multi-drug resistant multidrug-resistant Pseudomonas aeruginosa 60% ethanol extract of monkey earrings reduced ceftazidime MIC 50 from 32 ⁇ g/ml for single use to 8 ⁇ g/ml for 75% at a concentration of MIC; MIC 90 decreased from 256 ⁇ g/ml to 64 ⁇ g/ml, a 75% reduction;
  • the 60% ethanol extract of monkey earrings combined with amikacin FIC ⁇ 2 indicates that the two drugs have no antagonistic effect, 20% of FIC ⁇ 0.5 synergistic effect; for 20 multi-drug resistant multi-drug resistant Pseudomonas aeruginosa, Monkey earrings 60% ethanol extract reduced amikacin MIC 50 from 1 ⁇ g/ml for single use to 0.0625 ⁇ g/ml for 93.75% at a concentration of ⁇ MIC alone; MIC 90 decreased from 32 ⁇ g/ml to 2 ⁇ g /ml, reduced by 93.75%;
  • the combination of 60% ethanol extract of monkey earrings and imipenem after FIC ⁇ 2 indicates that the two drugs have no antagonistic effect, 15% of FIC ⁇ 0.5 synergistic effect; 20 strains of multi-drug resistant multi-drug resistant Pseudomonas aeruginosa
  • the monkey earrings 60% ethanol extract reduced the imipenem MIC 50 from 32 ⁇ g/ml for single use to 2 ⁇ g/ml at a concentration of ⁇ MIC, which was reduced by 93.75%; the MIC 90 decreased from 64 ⁇ g/ml to 16 ⁇ g. /ml, reduced by 75%;
  • the 60% ethanol extract of monkey earrings reduced the levofloxacin MIC 50 from 4 ⁇ g/ml to 1 ⁇ g/ml, at a concentration of ⁇ MIC alone, which was reduced by 75. %; MIC 90 decreased from 32 ⁇ g/ml to 8 ⁇ g/ml, a 75% decrease.
  • the MIC 50 and MIC 90 statistical analysis of the in vitro bacteriostasis and bactericidal action of monkey earrings 95% ethanol extract and five antibiotics against multidrug-resistant Pseudomonas aeruginosa are shown in Table P39.
  • Monkey earrings 95% ethanol extract alone has a MIC 50 of 800 ⁇ g/ml for multidrug-resistant Pseudomonas aeruginosa, 800 ⁇ g/ml for MIC 90, 1600 ⁇ g/ml for MBC 50 and 1600 ⁇ g/ml for MBC 90 ;
  • the combination of 95% ethanol extract of monkey earrings and imipenem has an FIC of ⁇ 2, indicating that the two drugs have no antagonistic effect, 40% of which has a synergistic effect of FIC ⁇ 0.5; and 20 multi-drug resistant multi-drug resistant Pseudomonas aeruginosa
  • the monkey earrings 95% ethanol extract reduced the imipenem MIC 50 from 32 ⁇ g/ml for single use to 4 ⁇ g/ml at a concentration of ⁇ MIC alone; the MIC 90 decreased from 64 ⁇ g/ml to 16 ⁇ g. /ml, reduced by 75%;
  • the 15% ethanol extract of monkey earrings reduced the levofloxacin MIC 50 from 4 ⁇ g/ml to 1 ⁇ g/ml at a concentration of ⁇ MIC alone. 75%; MIC 90 decreased from 32 ⁇ g/ml to 4 ⁇ g/ml, a decrease of 87.5%.
  • ESBL-producing Escherichia coli Anti-bacterial bactericidal test of monkey earring extract against extended-spectrum ⁇ -lactamase Escherichia coli (hereinafter referred to as ESBL-producing Escherichia coli) and sensitizing efficacy combined with amikacin and cotrimoxazole respectively Inspection.
  • the MIC 50 and MIC 90 statistical analysis of the in vitro antibacterial and bactericidal activity of monkey earring water extract and two antibiotics against ESBL-producing Escherichia coli are shown in Table E3.
  • the experimental results showed that the water extract of monkey earrings alone had an MIC 50 of 1600 ⁇ g/ml for ESBL-producing Escherichia coli, 1600 ⁇ g/ml for MIC 90, 3200 ⁇ g/ml for MBC 50, and 3200 ⁇ g/ml for MBC 90 ;
  • the FIC of monkey earring water extract combined with amikacin was ⁇ 2, indicating that there was no antagonistic effect of the two drugs, 50% of which had a FIC value of ⁇ 0.5, and the water extract of monkey earrings was
  • the MIC is less than or equal to 1/2 MIC
  • the MIC 50 of amikacin is reduced from 16 ⁇ g/ml used alone to 4 ⁇ g/ml, which is reduced by 75%
  • the MIC 90 is reduced from 64 ⁇ g/ml to 16 ⁇ g/ml, which is 75% lower;
  • the combination of monkey earring water extract and compound sulfamethoxazole on 20 strains of ESBL-producing Escherichia coli ⁇ 2 indicates that there is no antagonistic effect between the two drugs, and 25% FIC value ⁇ 0.5 is synergistic.
  • the monkey earring water extract reduced the compound sulfamethoxazole MIC 50 and MIC 90 from 1932/128 ⁇ g/ml for single use to 19/1 ⁇ g/ml at a ratio of 99.3% or less when the MIC was used alone or less.
  • the experimental results showed that the 10% ethanol extract of monkey earrings alone had an MIC 50 of 1600 ⁇ g/ml for ESBL-producing Escherichia coli, 1600 ⁇ g/ml for MIC 90, 3200 ⁇ g/ml for MBC 50, and 3200 ⁇ g/ml for MBC 90 ;
  • the combination of 10% ethanol extract of monkey earrings and compound sulfamethoxazole on 20 strains of ESBL-producing Escherichia coli ⁇ 2 showed no antagonistic effect of the two drugs, and 20% FIC value ⁇ 0.5 showed synergistic effect.
  • the monkey earrings 10% ethanol extract reduced the sulfamethoxazole MIC 50 and MIC 90 from 1932/128 ⁇ g/ml for single use to 19/1 ⁇ g/ml, which was reduced by 99.3%, when the MIC was less than or equal to MIC.
  • the MIC 50 and MIC 90 statistical analysis of the in vitro antibacterial and bactericidal activity of the monkey earrings 60% ethanol extract and two antibiotics against ESBL-producing Escherichia coli are shown in Table E21.
  • the experimental results show that the 60% ethanol extract of monkey earrings has an MIC 50 of 800 ⁇ g/ml for ESBL-producing Escherichia coli, 800 ⁇ g/ml for MIC 90, 1600 ⁇ g/ml for MBC 50 and 1600 ⁇ g/ml for MBC 90 ;
  • the 60% ethanol extract of monkey earrings combined with the compound sulfamethoxazole on the 20 strains of ESBL-producing Escherichia coli FIC ⁇ 2 indicates that the two drugs have no antagonistic effect, and 30% FIC value ⁇ 0.5 is synergistic.
  • the monkey earrings 60% ethanol extract reduced the sulfamethoxazole MIC 50 and MIC 90 from 1932/128 ⁇ g/ml for single use to 19/1 ⁇ g/ml, which was reduced by 99.3%, when the MIC was less than or equal to MIC.
  • the MIC 50 and MIC 90 statistical analysis of the in vitro antibacterial and bactericidal activity of monkey earrings 95% ethanol extract and two antibiotics against ESBL-producing Escherichia coli are shown in Table E30.
  • the 95% ethanol extract of monkey earrings was sterilized against ESBL-producing Escherichia coli.
  • the statistical analysis of MBC 50 and MBC 90 is shown in Table E31.
  • the experimental results showed that the 95% ethanol extract of monkey earrings alone had an MIC 50 of 1600 ⁇ g/ml for ESBL-producing Escherichia coli, 1600 ⁇ g/ml for MIC 90, 3200 ⁇ g/ml for MBC 50, and 3200 ⁇ g/ml for MBC 90 ;
  • the 95% ethanol extract of monkey earrings combined with Amikacin FIC ⁇ 2 indicates that there is no antagonistic effect of the two drugs, 50% of which have FIC value ⁇ 0.5 synergistic effect; monkey earrings 95% Ethanol extract reduced amikacin MIC 50 from 16 ⁇ g/ml used alone to 4 ⁇ g/ml at a ratio of 1/2 MIC, which was reduced by 75%; MIC 90 decreased from 64 ⁇ g/ml to 16 ⁇ g/ml. 75%;
  • the combination of 95% ethanol extract of monkey earrings and compound sulfamethoxazole on 20 strains of ESBL-producing Escherichia coli ⁇ 2 indicates that there is no antagonistic effect between the two drugs, and 20% FIC value ⁇ 0.5 is synergistic.
  • the monkey earrings 95% ethanol extract reduced the sulfamethoxazole MIC 50 and MIC 90 from 1932/128 ⁇ g/ml for single use to 19/1 ⁇ g/ml, which was 99.3% lower than the MIC alone.
  • Monkey earring extract is resistant to the antibacterial bactericidal test of extended-spectrum ⁇ -lactamase Klebsiella pneumoniae (hereinafter referred to as ESBL-producing Klebsiella pneumoniae).
  • the MIC 50 , MIC 90 , MBC 50 , and MBC 90 statistical analysis of the in vitro bacteriostasis and bactericidal action of monkey earring water extract on ESBLs-producing Klebsiella pneumoniae are shown in Table K2 and Table K3.
  • the MIC 50 , MIC 90 , MBC 50 and MBC 90 statistical analysis of the 10% ethanol extract of monkey earrings against the in vitro bacteriostasis and bactericidal activity of ESBLs-producing Klebsiella pneumoniae are shown in Table K5 and Table K6.

Landscapes

  • Health & Medical Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • Biotechnology (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Epidemiology (AREA)
  • Medical Informatics (AREA)
  • Botany (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Engineering & Computer Science (AREA)
  • Oncology (AREA)
  • Communicable Diseases (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

猴耳环水或乙醇提取物在制备抗耐药菌药物及与同类抗生素联用的增敏药物中的应用，所述耐药菌为多重耐药鲍曼不动杆菌、多重耐药铜绿假单胞菌、产超广谱β-内酰胺酶大肠杆菌、及产超广谱β-内酰胺酶肺炎克雷伯菌。

Description

一种中药提取物在制备抗菌药物中的应用 技术领域
本发明涉及猴耳环提取物的新用途，具体是猴耳环提取物在制备多种抗耐药菌药物中的应用。
背景技术
21世纪是多重耐药菌的时代，抗生素临床应用60年后，越来越多的院内感染和多重耐药菌感染成为目前临床抗菌治疗的一大难题，1961年Jerons报告第1例临床耐甲氧西林金黄色葡萄球菌(MRSA)感染患者，目前MRSA感染已逐渐遍及全球；2011年中国细菌耐药性监测临床中显示主要耐药菌的分布，大肠杆菌、肺炎克雷伯菌产超广谱β-内酰胺酶(ESBL)株分别为50.7％、38.5％，其耐药性的变迁和现状备受关注。另外据我国卫生部耐药监测协作组2010年度报告，鲍曼不动杆菌和铜绿假单胞菌在医院ICU分离的病原菌中居首位，鲍曼不动杆菌对亚胺培南和美罗培南的耐药率分别高达60.4％和61.4％。综上所述，在MRSA、产ESBL的各类细菌、多重耐药的鲍曼不动杆菌和铜绿假单胞菌的临床占有率逐年攀升，耐药率不断攀高的压力下，抗生素治疗面临着巨大的挑战。为避免细菌耐药现象的进一步恶化，专家学者力图发现新的抑制细菌生长以及治疗细菌引起疾病的新方法。已有研究报道证明中国传统中药如黄连、黄芩和连翘等对不同耐药细菌有一定的抑制效果，进一步研究中药抑制耐药菌生长的重点在于发现新的抑菌能力更强、耐药抑菌谱更广的中药。
猴耳环(Pithecellobium clypearia Benth)，学名围涎树，是含羞草科猴耳环属植物猴耳环的干燥幼枝和叶，其性味苦涩寒，功效清热解毒、 收湿敛疮，是治疗多种热毒症候独特的南方药材。
目前有文献公开猴耳环以及其提取物具有抗病毒作用。但缺乏猴耳环及其提取物在抗耐药菌方面的作用研究。
发明内容
为克服以上缺陷，本发明公开了猴耳环提取物在制备抗耐药菌及其与同类抗生素联用增敏作用方面的应用，具体为：
1、猴耳环提取物在制备抗多重耐药鲍曼不动杆菌药物或抗多重耐药鲍曼不动杆菌抗生素增敏药物中的应用。猴耳环提取物与抗生素亚胺培南或四环素或多粘菌素B或头孢他啶或左氧氟沙星联用，均体现明显的增敏作用。
2、猴耳环提取物在制备抗多重耐药铜绿假单胞菌药物或抗多重耐药铜绿假单胞菌抗生素增敏药物中的应用。猴耳环提取物与抗生素左氧氟沙星或亚胺培南或阿米卡星或头孢他啶或头孢哌酮联用，均体现明显的增敏作用。。
3、猴耳环提取物在制备抗产超广谱β-内酰胺酶大肠杆菌药物或抗产超广谱β-内酰胺酶大肠杆菌抗生素增敏药物中的应用。猴耳环提取物与抗生素阿米卡星或复方新诺明联用，均体现明显的增敏作用。
4、猴耳环提取物在制备抗产超广谱β-内酰胺酶肺炎克雷伯菌药物中的应用。猴耳环提取物对产超广谱β-内酰胺酶肺炎克雷伯菌体现明显的抑菌作用。
所述的猴耳环提取物优选猴耳环水提取物或猴耳环乙醇提取物。
猴耳环提取物的制备方法为：猴耳环粗粉用水或按体积比计浓度为 10％-95％的乙醇水溶液提取，所得的提取液再用乙酸乙酯萃取，所得的萃取物即为目标产品。所述的乙醇水溶液优选：按体积比计浓度为60％的乙醇水溶液。
本发明所称药物，包括人用药或动物用药，或土壤治理用药或相关制剂。
本发明的有益效果：
本发明首次公开了猴耳环提取物对多重耐药鲍曼不动杆菌、多重耐药铜绿假单胞菌、产超广谱β-内酰胺酶大肠杆菌、产超广谱β-内酰胺酶肺炎克雷伯菌的抗菌作用及其与该类抗生素联用的增敏作用。
经试验证明，猴耳环提取物与亚胺培南或四环素或多粘菌素B或头孢他啶或左氧氟沙星联用对抗多重耐药鲍曼不动杆菌的作用呈协同作用，较单用减少抗生素用量50％到87％。
猴耳环提取物与左氧氟沙星或亚胺培南或阿米卡星或头孢他啶或头孢哌酮联用对抗多重耐药铜绿假单胞菌的作用呈协同作用，较单用减少抗生素用量50％到99.2％。
猴耳环提取物与阿米卡星或复方新诺明联用对抗产超广谱β-内酰胺酶大肠杆菌的作用呈协同作用，较单用减少抗生素用量75％到99.3％。
本发明的猴耳环提取物可作为上述耐药菌天然的抗菌药物或其类似抗生素的增敏剂，应用于上述细菌引起的疾病治疗。本发明为解决该类抗生素的耐药性问题提供了新的途径和替代药物。适用于人用药、其他动物用药及对应感染菌的土壤治理。为天然植物提取物，无副作用，提取方法简单环保。
具体实施方式
本发明通过下述实施方案对猴耳环提取物抗耐药菌及对同类抗生素增敏效果的药理作用做进一步说明。
猴耳环提取物(EA)的制备：猴耳环(Pithecellobium clypearia Benth)由广州市花城制药厂提供。取适量猴耳环药材打成粗粉，用水或10％-95％的乙醇水溶液回流2次，每次2小时，过滤；合并滤液，浓缩得浸膏(即猴耳环水或乙醇提取物)。取浸膏用水混悬后，用乙酸乙酯萃取，萃取三次，合并乙酸乙酯萃取液，浓缩得乙酸乙酯萃取物。用10％乙醇水溶液回流所得提取物以下称猴耳环10％乙醇提取物，其他浓度乙醇水溶液回流所得提取物如此类推。
菌株：本发明的试验菌株，均由中山大学附属第一医院检验医学部临床微生物检验室提供，经中山大学附属第一医院临床微生物检验室检测确认其耐药性。
MH肉汤培养基：MH肉汤干粉(英国OXOID LTD.)2.1g，定容至100ml，NAOH调节pH至7.0，高压灭菌，置4℃冰箱备用。
供试品最低抑菌浓度(MIC)和最低杀菌浓度(MBC)的测定方法：以微量肉汤稀释法测定猴耳环水或乙醇提取物对多重耐药鲍曼不动杆菌的最低抑菌浓度(MIC)和最低杀菌浓度(MBC)。参照美国国立临床试验标准化委员会(NCCLS)推荐的微量肉汤稀释法操作。
供试品对同类抗生素增敏作用的考察方法：参照美国国立临床试验标准化委员会(NCCLS)推荐的棋盘稀释法操作。
一、猴耳环提取物抗多重耐药鲍曼不动杆菌的抑菌杀菌试验及分别与 亚胺培南、四环素、多粘菌素B、头孢他啶、左氧氟沙星联合增敏药效考察。
实施例1
1.实验方法
1)最低抑菌浓度(MIC)的测定：
猴耳环提取物、亚胺培南(IMP)、四环素(TE)、多粘菌素B(POLB)、头孢他啶(CAZ)、左氧氟沙星(LVX)分别在MH肉汤培养基中进行一系列倍比稀释，每孔50μl，调节接种菌为1.0×106CFU/ml，每孔加入50μl菌液。35℃培养；24小时，无沉淀出现的最低抗菌药物的浓度为其最低抑菌浓度(MIC)。
2)最低杀菌浓度(MBC)的测定：
采用平板涂布计数法，从1)项无菌生长的孔中吸取50ul菌悬液至血平板上，均匀涂布，35℃培养24小时，菌落计数，使最初的实验活菌数减少99.9％或以上所需要的最低抗菌药物的浓度为其最低杀菌浓度(MBC)。
通过测定药物的最低抑菌浓度(MIC)和最低杀菌浓度(MBC)，并对数据进行统计得出MIC50、MIC90、MBC50、MBC90，来评价药物抗多重耐药鲍曼不动杆菌的作用。
3)棋盘稀释法：
棋盘稀释法在96孔无菌培养板中进行，将猴耳环提取物及亚胺培南(IMP)、四环素(TE)、多粘菌素B(POLB)、头孢他啶(CAZ)分别在MH肉汤培养基中倍比稀释成系列浓度，以两药各自1/4MIC到4MIC分别进行联合，每孔A药B药各加25μL，调整菌悬液的浓度为1.0×106CFU/ml，每孔接种50μL菌液，35℃孵育24h后观察A药B药联合后对多重耐药鲍曼不 动杆菌的最低抑菌浓度(MIC)。
FIC指数的计算：
Figure PCTCN2017071671-appb-000001
通过计算部分抑菌指数(FIC)来评价猴耳环提取物与抗生素联合抗菌的相互作用，FIC≤0.5为协同作用，0.5﹤FIC≤1为相加作用，1﹤FIC≤2为无关作用，FIC﹥2为拮抗作用；并根据无菌生长孔找出猴耳环提取物和抗生素的最佳浓度配比，最终评价猴耳环提取物增强抗生素效力的作用。
本实施例的供试品包括：猴耳环水提取物、猴耳环10％乙醇提取物、猴耳环60％乙醇提取物、猴耳环95％乙醇提取物。
多重耐药鲍曼不动杆菌(MDRAB)，编号A1-A20。
2.实验结果
2.1猴耳环水提取物及五种抗生素(亚胺培南、四环素、多粘菌素B、头孢他啶、左氧氟沙星)对多重耐药鲍曼不动杆菌的体外抑菌试验结果见表A1。
猴耳环水提取物对多重耐药鲍曼不动杆菌的体外杀菌试验结果见表A2。
猴耳环水提取物及五种抗生素对多重耐药鲍曼不动杆菌的体外抑菌和杀菌的MIC50、MIC90统计分析见表A3。
猴耳环水提取物对多重耐药鲍曼不动杆菌杀菌MBC50、MBC90统计分析见表A4。
猴耳环水提取物与五种抗生素的联合药敏试验的FIC值及FIC值的分布统计结果见表A5、表A6。
猴耳环水提取物对五种抗生素的增敏作用及联用后MIC50、MIC90见表A7- 表A12。
表A1猴耳环水提取物及抗生素对多重耐药鲍曼不动杆菌体外抑菌结果
Figure PCTCN2017071671-appb-000002
表A2猴耳环水提取物对多重耐药鲍曼不动杆菌的体外杀菌试验结果
Figure PCTCN2017071671-appb-000003
表A3猴耳环水提取物及抗生素对MDRAB菌的体外抑菌MIC50、MIC90统计结果
Figure PCTCN2017071671-appb-000004
Figure PCTCN2017071671-appb-000005
表A4猴耳环水提取物对多重耐药鲍曼不动杆菌杀菌MBC50、MBC90
Figure PCTCN2017071671-appb-000006
表A5猴耳环水提取物与五种抗生素的联合药敏试验FIC值
Figure PCTCN2017071671-appb-000007
表A6猴耳环水提取物与五种抗生素的联合药敏试验FIC值分布统计
Figure PCTCN2017071671-appb-000008
Figure PCTCN2017071671-appb-000009
表A7猴耳环水提取物对亚胺培南的增敏作用及联用后MIC
Figure PCTCN2017071671-appb-000010
表A8猴耳环水提取物对四环素的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000011
Figure PCTCN2017071671-appb-000012
表A9猴耳环水提取物对多粘菌素B的增敏作用及联用后MIC
Figure PCTCN2017071671-appb-000013
表A10猴耳环水提取物对头孢他啶的增敏作用及联用后MIC
Figure PCTCN2017071671-appb-000014
Figure PCTCN2017071671-appb-000015
表A11猴耳环水提取物对左氧氟沙星的增敏作用及联用后MIC
Figure PCTCN2017071671-appb-000016
表A12联用后五种抗生素的MIC50、MIC90
Figure PCTCN2017071671-appb-000017
猴耳环水提取物单用对多重耐药鲍曼不动杆菌的MIC50＝600μg/ml，MI C90＝600μg/ml，MBC50＝1200μg/ml，MBC90＝1200μg/ml；
猴耳环水提取物与亚胺培南联用的FIC均≤1表明两药呈协同作用或部分协同作用，其中有35％FIC值≤0.5呈协同作用；对20株多重耐药鲍曼不动杆菌，猴耳环水提取物在浓度≤单用MIC时，使亚胺培南MIC50从单用的32μg/ml降至8μg/ml，降低了75％；MIC90从32μg/ml降至16μg/ml，降低50％；
猴耳环水提取物与四环素联用对20株多重耐药鲍曼不动杆菌FIC均≤1表明两药呈协同作用或部分协同作用，其中35％FIC值≤0.5呈协同作用；对20株多重耐药鲍曼不动杆菌，猴耳环水提取物在≤单用MIC时，使四环素MIC50单用256μg/ml降至64μg/ml，降低了75％；MIC90从512μg/ml降至128μg/ml，降低75％；
对20株多重耐药鲍曼不动杆菌，猴耳环水提取物与多粘菌素B联用FIC均≤2表明两药没有拮抗作用，其中25％FIC≤0.5呈协同作用；对20株多重耐药鲍曼不动杆菌，猴耳环水提取物在≤单用MIC时，使多粘菌素BMIC50单用4μg/ml降至1μg/ml，降低了75％，MIC90从16μg/ml降至4μg/ml，降低了75％；
猴耳环水提取物与头孢他啶联用后对20株多重耐药鲍曼不动杆菌FIC均≤2表明两药没有拮抗作用，其中20％FIC≤0.5呈协同作用；猴耳环水提取物可在≤单用MIC时，使头孢他啶MIC50从512μg/ml降至128μg/ml，下降了75％；MIC90从512μg/ml降至256μg/ml，下降了50％；
猴耳环水提取物与左氧氟沙星联用抗20株多重耐药鲍曼不动杆菌试验 FIC均≤2表明两药没有拮抗作用，其中有35％FIC值≤0.5表明两药有一定程度的协同作用；对20株多重耐药鲍曼不动杆菌，猴耳环水提取物在≤单用MIC时，使左氧氟沙星MIC50单用8μg/ml降至2μg/ml，降低了75％，MIC90从32μg/ml降至8μg/ml，降低了75％。
2.2猴耳环10％乙醇提取物及五种抗生素(亚胺培南、四环素、多粘菌素B、头孢他啶、左氧氟沙星)对多重耐药鲍曼不动杆菌的体外抑菌试验结果见表A13。
猴耳猴10％乙醇提取物对多重耐药鲍曼不动杆菌体外杀菌试验结果见表A14。
猴耳环10％乙醇提取物及五种抗生素对多重耐药鲍曼不动杆菌的体外抑菌和杀菌的MIC50、MIC90统计分析见表A15。
猴耳环提取物对多重耐药鲍曼不动杆菌杀菌MBC50、MBC90统计分析见表A16。
猴耳环10％乙醇提取物与五种抗生素的联合药敏试验的FIC值及FIC值的分布统计结果见表A17、表A18。
猴耳环10％乙醇提取物对五种抗生素的增敏作用及联用后其MIC50、MIC90见表A19-表A24。
表A13猴耳环10％乙醇提取物及五种抗生素对MDRAB体外抑菌试验结果
Figure PCTCN2017071671-appb-000018
Figure PCTCN2017071671-appb-000019
表A14猴耳环10％乙醇提取物对多重耐药鲍曼不动杆菌的体外杀菌试验结果
Figure PCTCN2017071671-appb-000020
表A15猴耳环10％乙醇提取物及抗生素对MDRAB体外抑菌MIC50、MIC90统计结果
Figure PCTCN2017071671-appb-000021
表A16猴耳环10％乙醇提取物对多重耐药鲍曼不动杆菌杀菌MBC50、MBC90
Figure PCTCN2017071671-appb-000022
表A17猴耳环10％乙醇提取物与五种抗生素的联合药敏试验的FIC值
Figure PCTCN2017071671-appb-000023
表A18猴耳环10％乙醇提取物与五种抗生素的联合药敏试验FIC值的分布统计
Figure PCTCN2017071671-appb-000024
表A19猴耳环10％乙醇提取物对亚胺培南的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000025
Figure PCTCN2017071671-appb-000026
表A20猴耳环10％乙醇提取物对四环素的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000027
表A21猴耳环10％乙醇提取物对多粘菌素B的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000028
Figure PCTCN2017071671-appb-000029
表A22猴耳环10％乙醇提取物对头孢他啶的增敏作用及联用后MIC
Figure PCTCN2017071671-appb-000030
表A23猴耳环10％乙醇提取物对左氧氟沙星的增敏作用及联用后MIC
Figure PCTCN2017071671-appb-000031
表A24联用后五种抗生素的MIC50、MIC90
Figure PCTCN2017071671-appb-000032
猴耳环10％乙醇提取物单用对多重耐药鲍曼不动杆菌的MIC50＝600μg/ml，MIC90＝600μg/ml，MBC50＝1200μg/ml，MBC90＝1200μg/ml；
猴耳环10％乙醇提取物与亚胺培南联用的FIC均≤1表明两药呈协同作用或部分协同作用，其中有45％FIC值≤0.5呈协同作用；对20株多重耐药鲍曼不动杆菌，猴耳环10％乙醇提取物在浓度≤1/2单用MIC时，使亚胺培 南MIC50从单用的32μg/ml降至8μg/ml，降低了75％；MIC90从32μg/ml降至16μg/ml，降低50％；
猴耳环10％乙醇提取物与四环素联用对20株多重耐药鲍曼不动杆菌FIC均≤1表明两药呈协同作用或部分协同作用，其中45％FIC值≤0.5呈协同作用；对20株多重耐药鲍曼不动杆菌，猴耳环10％乙醇提取物在小于等于1/2单用MIC时，使四环素MIC50单用256μg/ml降至64μg/ml，降低了75％；MIC90从512μg/ml降至256μg/ml，降低50％；
对20株多重耐药鲍曼不动杆菌，猴耳环10％乙醇提取物与多粘菌素B联用FIC均≤2表明两药没有拮抗作用，其中15％FIC≤0.5呈协同作用；对20株多重耐药鲍曼不动杆菌，猴耳环10％乙醇提取物在小于等于单用MIC时，使多粘菌素BMIC50单用4μg/ml降至1μg/ml，降低了75％，MIC90从16μg/ml降至4μg/ml，降低了75％；
猴耳环10％乙醇提取物与头孢他啶联用后对20株多重耐药鲍曼不动杆菌FIC均≤2表明两药没有拮抗作用，其中30％FIC≤0.5呈协同作用；猴耳环10％乙醇提取物的乙酸乙酯萃取物可在≤1/2单用MIC时，使头孢他啶MIC50从512μg/ml降至128μg/ml，下降了75％；MIC90从512μg/ml降至256μg/ml，下降了50％；
猴耳环10％乙醇提取物与左氧氟沙星联用抗20株多重耐药鲍曼不动杆菌试验FIC均≤2表明两药没有拮抗作用，其中有20％FIC值≤0.5表明两药有一定程度的协同作用；对20株多重耐药鲍曼不动杆菌，猴耳环10％乙醇提取物在小于等于单用MIC时，使左氧氟沙星MIC50单用8μg/ml降至4μg/ml，降低了50％，MIC90从32μg/ml降至8μg/ml，降低了75％。
2.3猴耳环60％乙醇提取物及五种抗生素(亚胺培南、四环素、多粘菌素B、头孢他啶、左氧氟沙星)对多重耐药鲍曼不动杆菌的体外抑菌试验结果见表A25。
猴耳环60％乙醇提取物对多重耐药鲍曼不动杆菌的体外杀菌试验结果见表A26。
猴耳环60％乙醇提取物及五种抗生素对多重耐药鲍曼不动杆菌的体外抑菌和杀菌的MIC50、MIC90统计分析见表A27。
猴耳环提取物对多重耐药鲍曼不动杆菌MBC50、MBC90统计分析见表28。
猴耳环60％乙醇提取物与五种抗生素的联合药敏试验的FIC值及FIC值的分布统计结果见表A29、表A30。
猴耳环60％乙醇提取物对五种抗生素的增敏作用及联用后其MIC50、MIC90见表A31-表A36。
表A25猴耳环60％乙醇提取物及五种抗生素对MDRAB体外抑菌试验结果
Figure PCTCN2017071671-appb-000033
Figure PCTCN2017071671-appb-000034
表A26猴耳环60％乙醇提取物对多重耐药鲍曼不动杆菌体外杀菌试验结果
Figure PCTCN2017071671-appb-000035
表A27猴耳环60％乙醇提取物及抗生素对MDRAB不动杆菌体外抑菌MIC50、MIC90统计
Figure PCTCN2017071671-appb-000036
表A28猴耳环60％乙醇提取物对多重耐药鲍曼不动杆菌的杀菌MBC50、MBC90
Figure PCTCN2017071671-appb-000037
表A29猴耳环60％乙醇提取物与五种抗生素的联合药敏试验的FIC值
Figure PCTCN2017071671-appb-000038
Figure PCTCN2017071671-appb-000039
表A30猴耳环60％乙醇提取物与五种抗生素的联合药敏试验FIC值的分布统计
Figure PCTCN2017071671-appb-000040
表A31猴耳环60％乙醇提取物对亚胺培南的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000041
Figure PCTCN2017071671-appb-000042
表A32猴耳环60％乙醇提取物对四环素的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000043
表A33猴耳环60％乙醇提取物对多粘菌素B的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000044
Figure PCTCN2017071671-appb-000045
表A34猴耳环60％乙醇提取物对头孢他啶的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000046
表A35猴耳环60％乙醇提取物对左氧氟沙星的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000047
Figure PCTCN2017071671-appb-000048
表A36联用后五种抗生素的MIC50、MIC90
Figure PCTCN2017071671-appb-000049
猴耳环60％乙醇提取物单用对多重耐药鲍曼不动杆菌的MIC50＝300μg/ml，MIC90＝600μg/ml，MBC50＝600μg/ml，MBC90＝1200μg/ml；
猴耳环60％乙醇提取物与亚胺培南联用的FIC均≤1表明两药呈协同作用或部分协同作用，其中有70％FIC值≤0.5呈协同作用；对20株多重耐药鲍曼不动杆菌，猴耳环60％乙醇提取物在浓度≤1/2单用MIC时，使亚胺培南MIC50从单用的32μg/ml降至8μg/ml，降低了75％；MIC90从32μg/ml降至16μg/ml，降低50％；
猴耳环60％乙醇提取物与四环素联用对20株多重耐药鲍曼不动杆菌FIC均≤1表明两药呈协同作用或部分协同作用，其中50％FIC值≤0.5呈协同作用；对20株多重耐药鲍曼不动杆菌，猴耳环60％乙醇提取物在小于等于1/2单用MIC时，使四环素MIC50单用256μg/ml降至32μg/ml，降低了87.5％；MIC90从512μg/ml降至128μg/ml，降低75％；
对20株多重耐药鲍曼不动杆菌，猴耳环60％乙醇提取物与多粘菌素B联用FIC均≤2表明两药没有拮抗作用，其中15％FIC≤0.5呈协同作用；对20株多重耐药鲍曼不动杆菌，猴耳环60％乙醇提取物在小于等于单用MIC时，使多粘菌素BMIC50单用4μg/ml降至0.5μg/ml，降低了87.5％，MIC90从16μg/ml降至4μg/ml，降低了75％；
猴耳环60％乙醇提取物与头孢他啶联用后对20株多重耐药鲍曼不动杆菌FIC均≤2表明两药没有拮抗作用，其中10％FIC≤0.5呈协同作用；猴耳环60％乙醇提取物可在≤1/2单用MIC时，使头孢他啶MIC50从512μg/ml降至128μg/ml，下降了75％；MIC90从512μg/ml降至256μg/ml，下降了50％；
猴耳环60％乙醇提取物与左氧氟沙星联用抗20株多重耐药鲍曼不动杆菌试验FIC均≤2表明两药没有拮抗作用，其中有5％FIC值≤0.5表明两药有一定程度的协同作用；对20株多重耐药鲍曼不动杆菌，猴耳环60％乙醇提取物在小于等于单用MIC时，使左氧氟沙星MIC50单用8μg/ml降至4μg/ml，降低了50％，MIC90从32μg/ml降至8μg/ml，降低了75％。
2.4猴耳环95％乙醇提取物及五种抗生素(亚胺培南、四环素、多粘菌素B、头孢他啶、左氧氟沙星)对多重耐药鲍曼不动杆菌的体外抑菌试验结果见表A37。
猴耳环95％乙醇提取物对多重耐药鲍曼不动杆菌体外杀菌试验结果见表A38。
猴耳环95％乙醇提取物及五种抗生素对多重耐药鲍曼不动杆菌的体外抑菌和杀菌的MIC50、MIC90统计分析见表A39。
猴耳环提取物对多重耐药鲍曼不动杆菌杀菌MBC50、MBC90统计分析见表A40。
猴耳环95％乙醇提取物与五种抗生素的联合药敏试验的FIC值及FIC值的分布统计结果见表A41、表A42。
猴耳环95％乙醇提取物对五种抗生素的增敏作用及联用后其MIC50、MIC90见表A43-表A48。
表A37猴耳环95％乙醇提取物及五种抗生素对MDRAB体外抑菌试验结果
Figure PCTCN2017071671-appb-000050
表A38猴耳环95％乙醇提取物对多重耐药鲍曼不动杆菌体外杀菌试验结果
Figure PCTCN2017071671-appb-000051
Figure PCTCN2017071671-appb-000052
表A39猴耳环95％乙醇提取物及抗生素对MDRAB体外抑菌MIC50、MIC90统计结果
Figure PCTCN2017071671-appb-000053
表A40猴耳环95％乙醇提取物对多重耐药鲍曼不动杆菌的杀菌MBC50、MBC90
Figure PCTCN2017071671-appb-000054
表A41猴耳环95％乙醇提取物与五种抗生素的联合药敏试验的FIC值
Figure PCTCN2017071671-appb-000055
Figure PCTCN2017071671-appb-000056
表A42猴耳环95％乙醇提取物与五种抗生素的联合药敏试验FIC值的分布统计
Figure PCTCN2017071671-appb-000057
表A43猴耳环95％乙醇提取物对亚胺培南的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000058
表A44猴耳环95％乙醇提取物对四环素的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000059
Figure PCTCN2017071671-appb-000060
表A45猴耳环95％乙醇提取物对多粘菌素B的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000061
表A46猴耳环95％乙醇提取物对头孢他啶的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000062
Figure PCTCN2017071671-appb-000063
表A47猴耳环95％乙醇提取物对左氧氟沙星的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000064
表A48联用后五种抗生素的MIC50、MIC90
Figure PCTCN2017071671-appb-000065
Figure PCTCN2017071671-appb-000066
猴耳环95％乙醇提取物单用对多重耐药鲍曼不动杆菌的MIC50＝600μg/ml，MIC90＝600μg/ml，MBC50＝1200μg/ml，MBC90＝1200μg/ml；
猴耳环95％乙醇提取物与亚胺培南联用的FIC均≤1表明两药呈协同作用或部分协同作用，其中有55％FIC值≤0.5呈协同作用；对20株多重耐药鲍曼不动杆菌，猴耳环95％乙醇提取物在浓度≤单用MIC时，使亚胺培南MIC50从单用的32μg/ml降至8μg/ml，降低了75％；MIC90从32μg/ml降至16μg/ml，降低50％；
猴耳环95％乙醇提取物与四环素联用对20株多重耐药鲍曼不动杆菌FIC均≤1表明两药呈协同作用或部分协同作用，其中30％FIC值≤0.5呈协同作用；对20株多重耐药鲍曼不动杆菌，猴耳环95％乙醇提取物在≤单用MIC时，使四环素MIC50单用256μg/ml降至32μg/ml，降低了87.5％；MIC90从512μg/ml降至128μg/ml，降低75％；
对20株多重耐药鲍曼不动杆菌，猴耳环95％乙醇提取物与多粘菌素B联用FIC均≤2表明两药没有拮抗作用，其中20％FIC≤0.5呈协同作用；对20株多重耐药鲍曼不动杆菌，猴耳环95％乙醇提取物在≤单用MIC时，使多粘菌素BMIC50单用4μg/ml降至1μg/ml，降低了75％，MIC90从16μg/ml降至4μg/ml，降低了75％；
猴耳环95％乙醇提取物与头孢他啶联用后对20株多重耐药鲍曼不动杆菌FIC均≤2表明两药没有拮抗作用，其中10％FIC≤0.5呈协同作用；猴 耳环95％乙醇提取物可在≤单用MIC时，使头孢他啶MIC50从512μg/ml降至128μg/ml，下降了75％；MIC90从512μg/ml降至256μg/ml，下降了50％；
猴耳环95％乙醇提取物与左氧氟沙星联用抗20株多重耐药鲍曼不动杆菌试验FIC均≤2表明两药没有拮抗作用，其中有20％FIC值≤0.5表明两药有一定程度的协同作用；对20株多重耐药鲍曼不动杆菌，猴耳环95％乙醇提取物在小于等于单用MIC时，使左氧氟沙星MIC50单用8μg/ml降至4μg/ml，降低了50％，MIC90从32μg/ml降至8μg/ml，降低了75％。
二、猴耳环提取物抗多重耐药铜绿假单胞菌的抑菌杀菌试验及分别与左氧氟沙星、亚胺培南、阿米卡星、头孢他啶、头孢哌酮联合的增敏药效考察。
实施例2
1.实验方法：多重耐药铜绿假单胞菌(MDRPA，编号P1-P20)菌株,参照本发明实施例1项下方法测定并评价。
2.实验结果：
2.1猴耳环水提取物及五种抗生素(左氧氟沙星、亚胺培南、阿米卡星、头孢他啶、头孢哌酮)对多重耐药铜绿假单胞菌的体外抑菌试验结果见表P1。
猴耳环水提取物对多重耐药铜绿假单胞菌体外杀菌试验结果见表P2。
猴耳环水提取物及五种抗生素对多重耐药铜绿假单胞菌的体外抑菌和杀菌的MIC50、MIC90统计分析见表P3。
猴耳环提取物对多重耐药铜绿假单胞菌MBC50、MBC90统计分析见表P4。
猴耳环水提取物与五种抗生素的联合药敏试验的FIC值及FIC值的分 布统计结果见表P5、表P6。
猴耳环水提取物对五种抗生素的增敏作用及联用后其MIC50、MIC90见表P7-表P12。
表P1猴耳环水提取物及五种抗生素对多重耐药铜绿假单胞菌体外抑菌试验结果
Figure PCTCN2017071671-appb-000067
表P2猴耳环水提取物对多重耐药铜绿假单胞菌的体外杀菌试验结果
Figure PCTCN2017071671-appb-000068
表P3猴耳环水提取物及五种抗生素对MDRPA体外抑菌MIC50、MIC90统计结果
Figure PCTCN2017071671-appb-000069
表P4猴耳环水提取物对多重耐药铜绿假单胞菌的杀菌的MBC50、MBC90
Figure PCTCN2017071671-appb-000070
表P5猴耳环水提取物与五种抗生素的联合药敏试验的FIC值
Figure PCTCN2017071671-appb-000071
表P6猴耳环水提取物与五种抗生素的联合药敏试验FIC值的分布统计结果
Figure PCTCN2017071671-appb-000072
Figure PCTCN2017071671-appb-000073
表P7猴耳环水提取物对头孢他啶的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000074
表P8猴耳环水提取物对头孢哌酮的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000075
Figure PCTCN2017071671-appb-000076
表P9猴耳环水提取物对阿米卡星的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000077
表P10猴耳环水提取物对亚胺培南的增敏作用及联用后MIC
Figure PCTCN2017071671-appb-000078
Figure PCTCN2017071671-appb-000079
表P11猴耳环水提取物对左氧氟沙星的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000080
表P12联用后五种抗生素的MIC50、MIC90
Figure PCTCN2017071671-appb-000081
猴耳环水提取物单用对多重耐药铜绿假单胞菌的MIC50为800μg/ml，MIC90为1600μg/ml，MBC50为1600μg/ml，MBC90为1600μg/ml；
对20株多重耐药铜绿假单胞菌，猴耳环水提取物与头孢他啶联用的FIC均≤2表明两药无拮抗作用，其中有45％FIC值≤0.5呈协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环水提取物在浓度≤单用MIC时，使头孢他啶MIC50从单用的32μg/ml降至8μg/ml，降低了75％；MIC90从256μg/ml降至64μg/ml，降低75％；
猴耳环水提取物与头孢哌酮联用对20株多重耐药铜绿假单胞菌FIC均≤2表明两药两药无拮抗作用，其中35％FIC值≤0.5呈协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环水提取物在浓度≤单用MIC时，使头孢哌酮MIC50从单用的64μg/ml降至0.5μg/ml，降低了99.2％；MIC90从512μg/ml降至256μg/ml，降低50％；
猴耳环水提取物与阿米卡星联用FIC均≤2表明两药没有拮抗作用，其中35％FIC≤0.5呈协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环水提取物在浓度≤单用MIC时，使阿米卡星MIC50从单用的1μg/ml降至0.0625μg/ml，降低了93.75％；MIC90从32μg/ml降至2μg/ml，降低93.75％；
猴耳环水提取物与亚胺培南联用后FIC均≤2表明两药没有拮抗作用，其中40％FIC≤0.5呈协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环水提取物在浓度≤单用MIC时，使亚胺培南MIC50从单用的32μg/ml降至2μg/ml，降低了93.75％；MIC90从64μg/ml降至16μg/ml，降低75％；
猴耳环水提取物与左氧氟沙星联用抗20株多重耐药铜绿假单胞菌，FIC 均≤2表明两药无拮抗作用，其中15％FIC值≤0.5表明两药有一定程度的协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环水提取物在浓度≤单用MIC时，使左氧氟沙星MIC50从单用的4μg/ml降至1μg/ml，降低了75％；MIC90从32μg/ml降至8μg/ml，降低75％。
2.2猴耳环10％乙醇提取物及五种抗生素(左氧氟沙星、亚胺培南、阿米卡星、头孢他啶、头孢哌酮)对多重耐药铜绿假单胞菌的体外抑菌试验结果见表P13。
猴耳环10％乙醇提取物对多重耐药铜绿假单胞菌的体外杀菌试验结果见表P14。
经过统计分析，猴耳环10％乙醇提取物及五种抗生素对多重耐药铜绿假单胞菌的体外抑菌和杀菌的MIC50、MIC90见表P15。
经过统计分析，猴耳环提取物对多重耐药铜绿假单胞菌的杀菌的MBC50、MBC90见表P16。
猴耳环10％乙醇提取物与五种抗生素的联合药敏试验的FIC值及FIC值的分布统计结果见表P17、表P18。
猴耳环10％乙醇提取物对五种抗生素的增敏作用及联用后其MIC50、MIC90见表P19-表P24。
表P13猴耳环10％乙醇提取物及五抗生素对MDRPA体外抑菌试验结果
Figure PCTCN2017071671-appb-000082
Figure PCTCN2017071671-appb-000083
表P14猴耳环10％乙醇提取物对多重耐药铜绿假单胞菌的体外杀菌试验结果
Figure PCTCN2017071671-appb-000084
表P15猴耳环10％乙醇提取物及抗生素对MDRPA体外抑菌MIC50、MIC90统计结果
Figure PCTCN2017071671-appb-000085
表P16猴耳环10％乙醇提取物对多重耐药铜绿假单胞菌的杀菌的MBC50、MBC90
Figure PCTCN2017071671-appb-000086
表P17猴耳环10％乙醇提取物与五种抗生素的联合药敏试验的FIC值
Figure PCTCN2017071671-appb-000087
表18猴耳环10％乙醇提取物与五种抗生素联合药敏试验FIC值分布统计结果
Figure PCTCN2017071671-appb-000088
表P19猴耳环10％乙醇提取物对头孢他啶的增敏作用及联用后MIC
Figure PCTCN2017071671-appb-000089
Figure PCTCN2017071671-appb-000090
表P20猴耳环10％乙醇提取物对头孢哌酮的增敏作用及联用后MIC
Figure PCTCN2017071671-appb-000091
Figure PCTCN2017071671-appb-000092
表P21猴耳环10％乙醇提取物对阿米卡星的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000093
表P22猴耳环10％乙醇提取物对亚胺培南的增敏作用及联用后MIC
Figure PCTCN2017071671-appb-000094
Figure PCTCN2017071671-appb-000095
表P23猴耳环10％乙醇提取物对左氧氟沙星的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000096
Figure PCTCN2017071671-appb-000097
表P24联用后五种抗生素的MIC50、MIC90
Figure PCTCN2017071671-appb-000098
猴耳环10％乙醇提取物单用对多重耐药铜绿假单胞菌的MIC50为800μg/ml，MIC90为1600μg/ml，MBC50为1600μg/ml，MBC90为1600μg/ml；
对20株多重耐药铜绿假单胞菌，猴耳环10％乙醇提取物与头孢他啶联用的FIC均≤2表明两药无拮抗作用，其中有35％FIC值≤0.5呈协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环10％乙醇提取物的在浓度≤单用MIC时，使头孢他啶MIC50从单用的32μg/ml降至16μg/ml，降低了50％；MIC90从256μg/ml降至64μg/ml，降低75％；
猴耳环10％乙醇提取物与头孢哌酮联用对20株多重耐药铜绿假单胞菌FIC均≤2表明两药两药无拮抗作用，其中35％FIC值≤0.5呈协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环10％乙醇提取物在浓度≤单用MIC时，使头孢哌酮MIC50从单用的64μg/ml降至4μg/ml，降低了93.75％；MIC90从512μg/ml降至256μg/ml，降低50％；
猴耳环10％乙醇提取物与阿米卡星联用FIC均≤2表明两药没有拮抗作用，其中30％FIC≤0.5呈协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环10％乙醇提取物在浓度≤单用MIC时，使阿米卡星MIC50从单用的1μg/ml降至0.0625μg/ml，降低了93.75％；MIC90从32μg/ml降至 4μg/ml，降低87.5％；
猴耳环10％乙醇提取物与亚胺培南联用后FIC均≤2表明两药没有拮抗作用，其中50％FIC≤0.5呈协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环10％乙醇提取物在浓度≤单用MIC时，使亚胺培南MIC50从单用的32μg/ml降至2μg/ml，降低了93.75％；MIC90从64μg/ml降至16μg/ml，降低75％；
猴耳环10％乙醇提取物与左氧氟沙星联用抗20株多重耐药铜绿假单胞菌，FIC均≤2表明两药无拮抗作用，其中15％FIC值≤0.5表明两药有一定程度的协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环15％乙醇提取物在浓度≤单用MIC时，使左氧氟沙星MIC50从单用的4μg/ml降至1μg/ml，降低了75％；MIC90从32μg/ml降至4μg/ml，降低87.5％。
2.3猴耳环60％乙醇提取物及五种抗生素(左氧氟沙星、亚胺培南、阿米卡星、头孢他啶、头孢哌酮)对多重耐药铜绿假单胞菌的体外抑菌试验结果见P表25。
猴耳环60％乙醇提取物对多重耐药铜绿假单胞菌体外杀菌试验结果见表P26。
猴耳环60％乙醇提取物及五种抗生素对多重耐药铜绿假单胞菌的体外抑菌和杀菌的MIC50、MIC90统计分析见表P27。
猴耳环提取物对多重耐药铜绿假单胞菌的杀菌的MBC50、MBC90统计分析见表P28。
猴耳环60％乙醇提取物与五种抗生素的联合药敏试验的FIC值及FIC值的分布统计结果见表P29、表P30。
猴耳环60％乙醇提取物对五种抗生素的增敏作用及联用后其MIC50、MIC90见表P31-表P36。
表P25猴耳环60％乙醇提取物及五种抗生素对MDRPA体外抑菌试验结果
Figure PCTCN2017071671-appb-000099
表P26猴耳环60％乙醇提取物对多重耐药铜绿假单胞菌的体外杀菌试验结果
Figure PCTCN2017071671-appb-000100
Figure PCTCN2017071671-appb-000101
表P27猴耳环60％乙醇提取物及抗生素对MDRPA的体外抑菌MIC50、MIC90统计结果
Figure PCTCN2017071671-appb-000102
表P28猴耳环60％乙醇提取物对多重耐药铜绿假单胞菌杀菌MBC50、MBC90
Figure PCTCN2017071671-appb-000103
表P29猴耳环60％乙醇提取物与五种抗生素的联合药敏试验FIC值
Figure PCTCN2017071671-appb-000104
Figure PCTCN2017071671-appb-000105
表P30猴耳环60％乙醇提取物与抗生素的联合药敏试验FIC值分布统计结果
Figure PCTCN2017071671-appb-000106
表P31猴耳环60％乙醇提取物对头孢他啶的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000107
表P32猴耳环60％乙醇提取物对头孢哌酮的增敏作用及联用后MIC
Figure PCTCN2017071671-appb-000108
Figure PCTCN2017071671-appb-000109
表P33猴耳环60％乙醇提取物对阿米卡星的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000110
Figure PCTCN2017071671-appb-000111
表P34猴耳环60％乙醇提取物对亚胺培南的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000112
表P35猴耳环60％乙醇提取物对左氧氟沙星的增敏作用及联用后MIC
Figure PCTCN2017071671-appb-000113
Figure PCTCN2017071671-appb-000114
表P36联用后五种抗生素的MIC50、MIC90
Figure PCTCN2017071671-appb-000115
猴耳环60％乙醇提取物单用对多重耐药铜绿假单胞菌的MIC50为400μg/ml，MIC90为800μg/ml，MBC50为1600μg/ml，MBC90为1600μg/ml；
对20株多重耐药铜绿假单胞菌，猴耳环60％乙醇提取物与头孢他啶联用的FIC均≤2表明两药无拮抗作用，其中有40％FIC值≤0.5呈协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环60％乙醇提取物在浓度≤单用MIC时，使头孢他啶MIC50从单用的32μg/ml降至8μg/ml，降低了75％； MIC90从256μg/ml降至64μg/ml，降低75％；
猴耳环60％乙醇提取物与头孢哌酮联用对20株多重耐药铜绿假单胞菌FIC均≤2表明两药两药无拮抗作用，其中20％FIC值≤0.5呈协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环60％乙醇提取物在浓度≤单用MIC时，使头孢哌酮MIC50从单用的64μg/ml降至4μg/ml，降低了93.75％；MIC90从512μg/ml降至256μg/ml，降低50％；
猴耳环60％乙醇提取物与阿米卡星联用FIC均≤2表明两药没有拮抗作用，其中20％FIC≤0.5呈协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环60％乙醇提取物在浓度≤单用MIC时，使阿米卡星MIC50从单用的1μg/ml降至0.0625μg/ml，降低了93.75％；MIC90从32μg/ml降至2μg/ml，降低93.75％；
猴耳环60％乙醇提取物与亚胺培南联用后FIC均≤2表明两药没有拮抗作用，其中15％FIC≤0.5呈协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环60％乙醇提取物在浓度≤单用MIC时，使亚胺培南MIC50从单用的32μg/ml降至2μg/ml，降低了93.75％；MIC90从64μg/ml降至16μg/ml，降低75％；
猴耳环60％乙醇提取物与左氧氟沙星联用抗20株多重耐药铜绿假单胞菌，有4株FIC大于2呈拮抗作用，其中10％FIC值≤0.5表明两药有一定程度的协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环60％乙醇提取物在浓度≤单用MIC时，使左氧氟沙星MIC50从单用的4μg/ml降至1μg/ml，降低了75％；MIC90从32μg/ml降至8μg/ml，降低75％。
2.4猴耳环95％乙醇提取物及五种抗生素(左氧氟沙星、亚胺培南、阿 米卡星、头孢他啶、头孢哌酮)对多重耐药铜绿假单胞菌的体外抑菌试验结果见表P37。
猴耳环95％乙醇提取物对多重耐药铜绿假单胞菌的体外杀菌试验结果见表P38。
猴耳环95％乙醇提取物及五种抗生素对多重耐药铜绿假单胞菌的体外抑菌和杀菌的MIC50、MIC90统计分析见表P39。
猴耳环提取物对多重耐药铜绿假单胞菌杀菌MBC50、MBC90统计分析见表P40。
猴耳环95％乙醇提取物与五种抗生素的联合药敏试验的FIC值及FIC值的分布统计结果见表P41、表P42。
猴耳环95％乙醇提取物对五种抗生素的增敏作用及联用后其MIC50、MIC90见表P43-表P48。
表P37猴耳环95％乙醇提取物及五种抗生素MDRPA体外抑菌试验结果
Figure PCTCN2017071671-appb-000116
Figure PCTCN2017071671-appb-000117
表P38猴耳环95％乙醇提取物对多重耐药铜绿假单胞菌的体外杀菌试验结果
Figure PCTCN2017071671-appb-000118
表P39猴耳环95％乙醇提取物及抗生素对MDRPA体外抑菌MIC50、MIC90统计结果
Figure PCTCN2017071671-appb-000119
表P40猴耳环95％乙醇提取物对多重耐药铜绿假单胞菌的杀菌MBC50、MBC90
Figure PCTCN2017071671-appb-000120
表P41猴耳环95％乙醇提取物与五种抗生素的联合药敏试验的FIC值
Figure PCTCN2017071671-appb-000121
Figure PCTCN2017071671-appb-000122
表P42猴耳环95％乙醇提取物与五种抗生素联合药敏试验FIC值分布统计结果
Figure PCTCN2017071671-appb-000123
表P43猴耳环10％乙醇提取物对头孢他啶的增敏作用及联用后MIC
Figure PCTCN2017071671-appb-000124
Figure PCTCN2017071671-appb-000125
表P44猴耳环95％乙醇提取物对头孢哌酮的增敏作用及联用后MIC
Figure PCTCN2017071671-appb-000126
表P45猴耳环95％乙醇提取物对阿米卡星的增敏作用及联用后的MIC
Figure PCTCN2017071671-appb-000127
Figure PCTCN2017071671-appb-000128
表P46猴耳环95％乙醇提取物对亚胺培南的增敏作用及联用后MIC
Figure PCTCN2017071671-appb-000129
表P47猴耳环95％乙醇提取物对左氧氟沙星的增敏作用及联用后MIC
Figure PCTCN2017071671-appb-000130
Figure PCTCN2017071671-appb-000131
表P48联用后五种抗生素的MIC50、MIC90
Figure PCTCN2017071671-appb-000132
猴耳环95％乙醇提取物单用对多重耐药铜绿假单胞菌的MIC50为800μg/ml，MIC90为800μg/ml，MBC50为1600μg/ml，MBC90为1600μg/ml；
对20株多重耐药铜绿假单胞菌，猴耳环95％乙醇提取物与头孢他啶联用的FIC均≤2表明两药无拮抗作用，其中有25％FIC值≤0.5呈协同作用；
对20株多重耐药多重耐药铜绿假单胞菌，猴耳环95％乙醇提取在浓度≤单用MIC时，使头孢他啶MIC50从单用的32μg/ml降至16μg/ml，降低了50％；MIC90从256μg/ml降至128μg/ml，降低50％；
猴耳环95％乙醇提取物与头孢哌酮联用对20株多重耐药铜绿假单胞菌FIC均≤2表明两药两药无拮抗作用，其中40％FIC值≤0.5呈协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环95％乙醇提取物在浓度≤单用MIC时，使头孢哌酮MIC50从单用的64μg/ml降至4μg/ml，降低了 93.75％；MIC90从512μg/ml降至128μg/ml，降低75％；
猴耳环95％乙醇提取物与阿米卡星联用FIC均≤2表明两药没有拮抗作用，其中40％FIC≤0.5呈协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环95％乙醇提取物在浓度≤单用MIC时，使阿米卡星MIC50从单用的1μg/ml降至0.12μg/ml，降低了87.5％；MIC90从32μg/ml降至4μg/ml，降低87.5％；
猴耳环95％乙醇提取物与亚胺培南联用后FIC均≤2表明两药没有拮抗作用，其中40％FIC≤0.5呈协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环95％乙醇提取物在浓度≤单用MIC时，使亚胺培南MIC50从单用的32μg/ml降至4μg/ml，降低了75％；MIC90从64μg/ml降至16μg/ml，降低75％；
猴耳环95％乙醇提取物与左氧氟沙星联用抗20株多重耐药铜绿假单胞菌，FIC均≤2表明两药无拮抗作用，其中10％FIC值≤0.5表明两药有一定程度的协同作用；对20株多重耐药多重耐药铜绿假单胞菌，猴耳环15％乙醇提取物在浓度≤单用MIC时，使左氧氟沙星MIC50从单用的4μg/ml降至1μg/ml，降低了75％；MIC90从32μg/ml降至4μg/ml，降低87.5％。
三、猴耳环提取物抗产超广谱β-内酰胺酶大肠杆菌(以下简称：产ESBL大肠杆菌)的抑菌杀菌试验及分别与阿米卡星、复方新诺明联合的增敏药效考察。
实施例3
1.实验方法：产超广谱β-内酰胺酶大肠杆菌(ECO，编号E1-E20)菌株；参照本发明实施例1项下方法测定并评价。
2.实验结果：
2.1猴耳环水提取物及两种抗生素(阿米卡星、复方新诺明)对产ESBL大肠杆菌的体外抑菌试验结果见表E1。
猴耳环水提取物对产ESBLs大肠杆菌的体外杀菌试验结果见表E2。
猴耳环水提取物及两种抗生素对产ESBL大肠杆菌的体外抑菌和杀菌的MIC50、MIC90统计分析见表E3。
猴耳环提取物对产ESBL大肠杆菌的杀菌的MBC50、MBC90统计分析见表E4。
猴耳环水提取物与两种抗生素的联合药敏试验的FIC值及FIC值的分布统计结果见表E5、表E6。
猴耳环水提取物对两种抗生素的增敏作用及联用后其MIC50、MIC90见表E7-E9。
表E1
Figure PCTCN2017071671-appb-000133
Figure PCTCN2017071671-appb-000134
表E2
Figure PCTCN2017071671-appb-000135
表E3
Figure PCTCN2017071671-appb-000136
表E4
Figure PCTCN2017071671-appb-000137
表E5
Figure PCTCN2017071671-appb-000138
Figure PCTCN2017071671-appb-000139
表E6
Figure PCTCN2017071671-appb-000140
表E7
Figure PCTCN2017071671-appb-000141
表E8
Figure PCTCN2017071671-appb-000142
表E9
Figure PCTCN2017071671-appb-000143
实验结果表明猴耳环水提取物单用对产ESBL大肠杆菌的MIC50为1600μg/ml，MIC90为1600μg/ml，MBC50为3200μg/ml，MBC90为3200μg/ml；
对20株产ESBL大肠杆菌，猴耳环水提取物与阿米卡星联用的FIC均≤2表明两药无拮抗作用，其中有50％FIC值≤0.5呈协同作用；猴耳环水提取物在小于等于1/2MIC时使阿米卡星MIC50从单用的16μg/ml降至4μg/ml，降低了75％；MIC90从单用64μg/ml降至16μg/ml，降低75％；
猴耳环水提取物与复方新诺明联用对20株产ESBL大肠杆菌FIC均≤2表明两药两药无拮抗作用，其中25％FIC值≤0.5呈协同作用。猴耳环水提取物在小于等于单用MIC时使复方新诺明MIC50和MIC90均从单用的2432/128μg/ml降至19/1μg/ml，降低了99.3％。
2.2猴耳环10％乙醇提取物及两种抗生素(阿米卡星、复方新诺明)对产ESBL大肠杆菌的体外抑菌试验结果见表E10。
猴耳环10％乙醇提取物对产ESBLs大肠杆菌的体外杀菌试验结果见表E11。
猴耳环10％乙醇提取物及两种抗生素对产ESBL大肠杆菌的体外抑菌和杀菌MIC50、MIC90统计分析见表E12。
猴耳环10％乙醇提取物对产ESBL大肠杆菌的杀菌的MBC50、MBC90统计分析见表E13。
猴耳环10％乙醇提取物与两种抗生素的联合药敏试验的FIC值及FIC值的分布统计结果见表E14、表E15。
猴耳环10％乙醇提取物对两种抗生素的增敏作用及联用后其MIC50、MIC90见表E16-E18。
表E10
Figure PCTCN2017071671-appb-000144
Figure PCTCN2017071671-appb-000145
表E11
Figure PCTCN2017071671-appb-000146
表E12
Figure PCTCN2017071671-appb-000147
表E13
Figure PCTCN2017071671-appb-000148
表E14
Figure PCTCN2017071671-appb-000149
Figure PCTCN2017071671-appb-000150
表E15
Figure PCTCN2017071671-appb-000151
表E16
Figure PCTCN2017071671-appb-000152
Figure PCTCN2017071671-appb-000153
表E17
Figure PCTCN2017071671-appb-000154
表E18
Figure PCTCN2017071671-appb-000155
实验结果表明猴耳环10％乙醇提取物单用对产ESBL大肠杆菌的MIC50为1600μg/ml，MIC90为1600μg/ml，MBC50为3200μg/ml，MBC90为3200μg/ml；
对20株产ESBL大肠杆菌，猴耳环10％乙醇提取物与阿米卡星联用的FIC均≤2表明两药无拮抗作用，其中有50％FIC值≤0.5呈协同作用；猴 耳环10％乙醇提取物在小于等于1/2MIC时使阿米卡星MIC50从单用的16μg/ml降至4μg/ml，降低了75％；MIC90从单用64μg/ml降至16μg/ml，降低75％；
猴耳环10％乙醇提取物与复方新诺明联用对20株产ESBL大肠杆菌FIC均≤2表明两药两药无拮抗作用，其中20％FIC值≤0.5呈协同作用。猴耳环10％乙醇提取物在小于等于单用MIC时使复方新诺明MIC50和MIC90均从单用的2432/128μg/ml降至19/1μg/ml，降低了99.3％。
2.3猴耳环60％乙醇提取物及两种抗生素(阿米卡星、复方新诺明)对产ESBL大肠杆菌的体外抑菌试验结果见表E19。
猴耳环60％乙醇提取物对产ESBLs大肠杆菌的体外杀菌试验结果见表E20。
猴耳环60％乙醇提取物及两种抗生素对产ESBL大肠杆菌的体外抑菌和杀菌的MIC50、MIC90统计分析见表E21。
猴耳环60％乙醇提取物对产ESBL大肠杆菌的杀菌的MBC50、MBC90统计分析见表E22。
猴耳环60％乙醇提取物与两种抗生素的联合药敏试验的FIC值及FIC值的分布统计结果见表E23、表E24。
猴耳环60％乙醇提取物对两种抗生素的增敏作用及联用后其MIC50、MIC90见表E25-E27。
表E19
Figure PCTCN2017071671-appb-000156
Figure PCTCN2017071671-appb-000157
表E20
Figure PCTCN2017071671-appb-000158
表E21
Figure PCTCN2017071671-appb-000159
表E22
Figure PCTCN2017071671-appb-000160
表E23
Figure PCTCN2017071671-appb-000161
表E24
Figure PCTCN2017071671-appb-000162
表E25
Figure PCTCN2017071671-appb-000163
Figure PCTCN2017071671-appb-000164
表E26
Figure PCTCN2017071671-appb-000165
表E27
Figure PCTCN2017071671-appb-000166
实验结果表明猴耳环60％乙醇提取物单用对产ESBL大肠杆菌的MIC50为800μg/ml，MIC90为800μg/ml，MBC50为1600μg/ml，MBC90为1600μg/ml；
对20株产ESBL大肠杆菌，猴耳环60％乙醇提取物与阿米卡星联用的FIC均≤2表明两药无拮抗作用，其中有70％FIC值≤0.5呈协同作用；猴耳环60％乙醇提取物在小于等于1/4MIC时使阿米卡星MIC50从单用的16μg/ml降至4μg/ml，降低了75％；MIC90从单用64μg/ml降至8μg/ml；
猴耳环60％乙醇提取物与复方新诺明联用对20株产ESBL大肠杆菌FIC均≤2表明两药两药无拮抗作用，其中30％FIC值≤0.5呈协同作用。猴耳环60％乙醇提取物在小于等于单用MIC时使复方新诺明MIC50和MIC90均从单用的2432/128μg/ml降至19/1μg/ml，降低了99.3％。
2.4猴耳环95％乙醇提取物及两种抗生素(阿米卡星、复方新诺明)对产ESBL大肠杆菌的体外抑菌试验结果见表E28。
猴耳环95％乙醇提取物对产ESBLs大肠杆菌的体外杀菌试验结果见表E29。
猴耳环95％乙醇提取物及两种抗生素对产ESBL大肠杆菌的体外抑菌和杀菌的MIC50、MIC90统计分析见表E30。
猴耳环95％乙醇提取物对产ESBL大肠杆菌杀菌MBC50、MBC90统计分析见表E31。
猴耳环95％乙醇提取物与两种抗生素的联合药敏试验的FIC值及FIC值的分布统计结果见表E32、表E33。
猴耳环95％乙醇提取物对两种抗生素的增敏作用及联用后其MIC50、MIC90见表E34-E36。
表E28
Figure PCTCN2017071671-appb-000167
Figure PCTCN2017071671-appb-000168
表E29
Figure PCTCN2017071671-appb-000169
表E30
Figure PCTCN2017071671-appb-000170
表E31
Figure PCTCN2017071671-appb-000171
表E32
Figure PCTCN2017071671-appb-000172
表E33
Figure PCTCN2017071671-appb-000173
表E34
Figure PCTCN2017071671-appb-000174
Figure PCTCN2017071671-appb-000175
表E35
Figure PCTCN2017071671-appb-000176
表E36
Figure PCTCN2017071671-appb-000177
Figure PCTCN2017071671-appb-000178
实验结果表明猴耳环95％乙醇提取物单用对产ESBL大肠杆菌的MIC50为1600μg/ml，MIC90为1600μg/ml，MBC50为3200μg/ml，MBC90为3200μg/ml；
对20株产ESBL大肠杆菌，猴耳环95％乙醇提取物与阿米卡星联用的FIC均≤2表明两药无拮抗作用，其中有50％FIC值≤0.5呈协同作用；猴耳环95％乙醇提取物在小于等于1/2MIC时使阿米卡星MIC50从单用的16μg/ml降至4μg/ml，降低了75％；MIC90从单用64μg/ml降至16μg/ml，降低75％；
猴耳环95％乙醇提取物与复方新诺明联用对20株产ESBL大肠杆菌FIC均≤2表明两药两药无拮抗作用，其中20％FIC值≤0.5呈协同作用。猴耳环95％乙醇提取物在小于等于单用MIC时使复方新诺明MIC50和MIC90均从单用的2432/128μg/ml降至19/1μg/ml，降低了99.3％。
四、猴耳环提取物抗产超广谱β-内酰胺酶肺炎克雷伯菌(以下称：产ESBL肺炎克雷伯菌)的抑菌杀菌试验。
实施例4
1.实验方法：产ESBL肺炎克雷伯菌(KPN，编号K1-K20)菌株；参照本发明实施例1项下抑菌杀菌试验方法测定并评价。
2.实验结果：
猴耳环水提取物对产ESBL肺炎克雷伯菌的体外抑菌杀菌试验结果见表K1。
猴耳环水提取物对产ESBLs肺炎克雷伯菌的体外抑菌和杀菌的MIC50、 MIC90、MBC50、MBC90统计分析见表K2、表K3。
猴耳环10％乙醇提取物对产ESBL肺炎克雷伯菌的体外抑菌杀菌试验结果见表K4。
猴耳环10％乙醇提取物对产ESBLs肺炎克雷伯菌的体外抑菌和杀菌的MIC50、MIC90、MBC50、MBC90统计分析见表K5、表K6。
猴耳环60％乙醇提取物对产ESBL肺炎克雷伯菌的体外抑菌杀菌试验结果见表K7。
猴耳环60％乙醇提取物对产ESBLs肺炎克雷伯菌的体外抑菌和杀菌的MIC50、MIC90、MBC50、MBC90统计分析见表K8、表K9。
猴耳环95％乙醇提取物对产ESBL肺炎克雷伯菌的体外抑菌杀菌试验结果见表K10。
猴耳环95％乙醇提取物对产ESBLs肺炎克雷伯菌的体外抑菌和杀菌的MIC50、MIC90、MBC50、MBC90统计分析见表K11、表K12。
表K1猴耳环水提取物对产ESBL肺炎克雷伯菌体外抑菌杀菌试验结果
Figure PCTCN2017071671-appb-000179
Figure PCTCN2017071671-appb-000180
表K2猴耳环水提取物对产ESBL肺炎克雷伯菌的体外抑菌MIC50、MIC90
Figure PCTCN2017071671-appb-000181
表K3猴耳环水提取物对产ESBL肺炎克雷伯菌的体外抑菌MBC50、MBC90
Figure PCTCN2017071671-appb-000182
表K4猴耳环10％乙醇提取物对产ESBL肺炎克雷伯菌体外抑菌杀菌试验结果
Figure PCTCN2017071671-appb-000183
表K5猴耳环10％乙醇提取物对产ESBL肺炎克雷伯菌体外抑菌MIC50、MIC90
Figure PCTCN2017071671-appb-000184
表K6猴耳环10％乙醇提取物对产ESBL肺炎克雷伯菌体外抑菌MBC50、MBC90
Figure PCTCN2017071671-appb-000185
表K7猴耳环60％乙醇提取物对产ESBL肺炎克雷伯菌体外抑菌杀菌试验结果
Figure PCTCN2017071671-appb-000186
表K8猴耳环60％乙醇提取物对产ESBL肺炎克雷伯菌体外抑菌MIC50、MIC90
Figure PCTCN2017071671-appb-000187
表9猴耳环60％乙醇提取物对产ESBL肺炎克雷伯菌体外抑菌MBC50、MBC90
Figure PCTCN2017071671-appb-000188
表K10猴耳环95％乙醇提取物对产ESBL肺炎克雷伯菌体外抑菌杀菌试验结果
Figure PCTCN2017071671-appb-000189
Figure PCTCN2017071671-appb-000190
表K11猴耳环95％乙醇提取物对产ESBL肺炎克雷伯菌体外抑菌MIC50、MIC90
Figure PCTCN2017071671-appb-000191
表K12猴耳环95％乙醇提取物对产ESBL肺炎克雷伯菌体外抑菌MBC50、MBC90
Figure PCTCN2017071671-appb-000192
猴耳环水提取物单用对产ESBL肺炎克雷伯菌的MIC50＝1600μg/ml，MIC90＝1600μg/ml，MBC50及MBC90均大于1600μg/ml。
猴耳环10％乙醇提取物单用对产ESBL肺炎克雷伯菌的MIC50＝1600μg/ml，MIC90＝1600μg/ml，MBC50及MBC90均大于1600μg/ml。
猴耳环60％乙醇提取物单用对产ESBL肺炎克雷伯菌的MIC50＝800μg/ml，MIC90＝1600μg/ml，MBC50及MBC990均大于1600μg/ml。
猴耳环95％乙醇提取物单用对产ESBL肺炎克雷伯菌的MIC50＝1600μg/ml，MIC90＝1600μg/ml，MBC50及MBC90均大于1600μg/ml。

Claims (10)

  1. 猴耳环提取物在制备抗多重耐药鲍曼不动杆菌药物或抗多重耐药鲍曼不动杆菌抗生素增敏药物中的应用。
  2. 如权利要求1所述的应用，其特征在于，所述的抗生素是亚胺培南或四环素或多粘菌素B或头孢他啶或左氧氟沙星。
  3. 猴耳环提取物在制备抗多重耐药铜绿假单胞菌药物或抗多重耐药铜绿假单胞菌抗生素增敏药物中的应用。
  4. 如权利要求3所述的应用，其特征在于，所述的抗生素是左氧氟沙星或亚胺培南或阿米卡星或头孢他啶或头孢哌酮。
  5. 猴耳环提取物在制备抗产超广谱β-内酰胺酶大肠杆菌药物或抗产超广谱β-内酰胺酶大肠杆菌抗生素增敏药物中的应用。
  6. 如权利要求5所述的应用，其特征在于，所述的抗生素是阿米卡星或复方新诺明。
  7. 猴耳环提取物在制备抗产超广谱β-内酰胺酶肺炎克雷伯菌药物中的应用。
  8. 如权利要求1到7所述的应用，其特征在于，所述的猴耳环提取物是猴耳环水提取物或猴耳环乙醇提取物。
  9. 如权利要求8所述的应用，其特征在于，所述的猴耳环水提取物水或猴耳环乙醇提取物由以下方法制备：猴耳环粗粉用水或按体积比计浓度为10％-95％的乙醇水溶液提取，所得的提取液再用乙酸乙酯萃取，所得的萃取物即为目标产品。
  10. 权利要求9所述的应用，其特征在于，所述的乙醇水溶液是按体积比计浓度为60％的乙醇水溶液。
PCT/CN2017/071671 2016-05-10 2017-01-19 猴耳环提取物在制备抗菌药物中的应用 WO2017193635A1 (zh)

Priority Applications (7)

Application Number Priority Date Filing Date Title
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
US17/494,858 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
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

Applications Claiming Priority (8)

Application Number Priority Date Filing Date Title
CN201610308716.7 2016-05-10
CN201610308718.6 2016-05-10
CN201610308717.1 2016-05-10
CN201610308716.7A CN105963339B (zh) 2016-05-10 2016-05-10 猴耳环提取物在制备抗多重耐药铜绿假单胞菌药物中的应用
CN201610308207.4 2016-05-10
CN201610308718.6A CN105998153B (zh) 2016-05-10 2016-05-10 猴耳环提取物在制备抗产超广谱β-内酰胺酶大肠杆菌药物中的应用
CN201610308717.1A CN105816511B (zh) 2016-05-10 2016-05-10 猴耳环提取物在制备抗多重耐药鲍曼不动杆菌药物中的应用
CN201610308207.4A CN105920082B (zh) 2016-05-10 2016-05-10 猴耳环提取物在制备抗产超广谱β-内酰胺酶肺炎克雷伯菌药物中的应用

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US15/920,480 Continuation 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

Publications (2)

Publication Number Publication Date
WO2017193635A1 true WO2017193635A1 (zh) 2017-11-16
WO2017193635A9 WO2017193635A9 (zh) 2017-12-14

Family

ID=60267889

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2017/071671 WO2017193635A1 (zh) 2016-05-10 2017-01-19 猴耳环提取物在制备抗菌药物中的应用

Country Status (3)

Country Link
US (6) US11154582B2 (zh)
EP (1) EP3456335B1 (zh)
WO (1) WO2017193635A1 (zh)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114129588B (zh) * 2021-10-28 2024-01-23 湖南农业大学 含黄芪提取物的抗菌组合物及其应用
CN114042100B (zh) * 2021-10-28 2023-05-02 湖南农业大学 含中药提取物的抗菌组合物及其应用

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1718222A (zh) * 2004-07-06 2006-01-11 广州莱泰制药有限公司 一种消炎口含片
CN103385912A (zh) * 2013-07-24 2013-11-13 中山大学 猴耳环提取物及其在制备抗耐甲氧西林金黄色葡萄球菌药物中的应用
CN105816511A (zh) * 2016-05-10 2016-08-03 中山大学 猴耳环提取物在制备抗多重耐药鲍曼不动杆菌药物中的应用
CN105920082A (zh) * 2016-05-10 2016-09-07 中山大学 猴耳环提取物在制备抗产超广谱β-内酰胺酶肺炎克雷伯菌药物中的应用
CN105963339A (zh) * 2016-05-10 2016-09-28 中山大学 猴耳环提取物在制备抗多重耐药铜绿假单胞菌药物中的应用
CN105998153A (zh) * 2016-05-10 2016-10-12 中山大学 猴耳环提取物在制备抗产超广谱β-内酰胺酶大肠杆菌药物中的应用

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101032547A (zh) 2006-03-07 2007-09-12 广东奇方药业有限公司 一种抗炎抗病毒的药物组合物
CN103860627A (zh) * 2012-12-10 2014-06-18 广东高山动物药业有限公司 一种猴耳环散抗菌兽药

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1718222A (zh) * 2004-07-06 2006-01-11 广州莱泰制药有限公司 一种消炎口含片
CN103385912A (zh) * 2013-07-24 2013-11-13 中山大学 猴耳环提取物及其在制备抗耐甲氧西林金黄色葡萄球菌药物中的应用
CN105816511A (zh) * 2016-05-10 2016-08-03 中山大学 猴耳环提取物在制备抗多重耐药鲍曼不动杆菌药物中的应用
CN105920082A (zh) * 2016-05-10 2016-09-07 中山大学 猴耳环提取物在制备抗产超广谱β-内酰胺酶肺炎克雷伯菌药物中的应用
CN105963339A (zh) * 2016-05-10 2016-09-28 中山大学 猴耳环提取物在制备抗多重耐药铜绿假单胞菌药物中的应用
CN105998153A (zh) * 2016-05-10 2016-10-12 中山大学 猴耳环提取物在制备抗产超广谱β-内酰胺酶大肠杆菌药物中的应用

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
SHA, JINGSHU ET AL.: "Pithecellobium Clypearia", CHINESE PHARMACEUTICAL BULLETIN, vol. 21, no. 11, 30 November 1986 (1986-11-30), pages 665, XP009510155 *

Also Published As

Publication number Publication date
EP3456335A1 (en) 2019-03-20
US11654174B2 (en) 2023-05-23
EP3456335A4 (en) 2019-05-08
US20220031789A1 (en) 2022-02-03
US11793849B2 (en) 2023-10-24
EP3456335B1 (en) 2020-10-14
US20210106641A1 (en) 2021-04-15
US11154582B2 (en) 2021-10-26
US20220023369A1 (en) 2022-01-27
US11491198B2 (en) 2022-11-08
US20210106639A1 (en) 2021-04-15
US20180200319A1 (en) 2018-07-19
WO2017193635A9 (zh) 2017-12-14
US20210106640A1 (en) 2021-04-15

Similar Documents

Publication Publication Date Title
Kambiz et al. Extracts from Aloe ferox and Withania somnifera inhibit Candida albicans and Neisseria gonorrhoea
Bag et al. Evaluation of antibacterial properties of Chebulic myrobalan (fruit of Terminalia chebula Retz.) extracts against methicillin resistant Staphylococcus aureus and trimethoprim-sulphamethoxazole resistant uropathogenic Escherichia coli
US11793849B2 (en) Method of Chinese herbal medicine extract used for treating multiple diseases caused by drug resistant bacteria infection
Shahwany et al. Antibacterial and anti-biofilm activity of three phenolic plant extracts and silver nanoparticles on Staphylococcus aureus and Klebsiella pneumoniae
CN105816511B (zh) 猴耳环提取物在制备抗多重耐药鲍曼不动杆菌药物中的应用
Coopoosamy et al. Assessing the potential of Tetradenia riparia in treatment of common skin conditions in rural communities of South Africa
CN105920082B (zh) 猴耳环提取物在制备抗产超广谱β-内酰胺酶肺炎克雷伯菌药物中的应用
Al-Hayanni The Antibacterial and anti-biofilm effects of Sumac (Rhus coriaria L) fruits extracts against some multidrug-resistant pathogenic bacteria
CN111701011A (zh) 宿主防御肽组合物及其在制备抗菌药物中的应用
Fayyad et al. Evaluation of synergistic effect of nicotinic acid with imipenem as antibiofilm for clinical Pseudomonas Aeruginosa isolates
CN113082026B (zh) 一种青蒿素衍生物在制备多粘菌素抗菌增效剂中的应用
CN110974819B (zh) 一种用于上呼吸道感染的组合物及其应用
Rajesh et al. Evaluation of antimicrobial activity of different herbal plant extracts
Goudarzi et al. Antimicrobial activity of Peganum harmala against methicillin-resistant Staphylococcus aureus strains and assessment of its cytotoxicity effect on HEK-293 cells
CN113855669A (zh) 一种抗菌的联合用药物
WO2002022147A1 (en) Antibacterial combination comprising neem plant extract
Verma et al. Antibacterial properties of seabuckthorn (Hippophae rhamnoides L.) leaf extracts against common skin and wound bacteria
PL221245B1 (pl) Zastosowanie olejku eterycznego z lawendy wąskolistnej odmiany ,,Blue River"
JP2023519453A (ja) 青銭柳葉フラボン抽出物の抗菌薬物および／または抗菌剤の製造における使用
CN110772518A (zh) 血根碱在抑制路邓葡萄球菌生长中的应用
Haque et al. Antibacterial effect of Dimethyl sulfoxide extract of Aloe vera (Aloe barbadensis) leaf gel against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae
CN110237058B (zh) 柠檬醛在抑制多重耐药霍氏肠杆菌生长中的应用
KR102330947B1 (ko) 하고초 추출물을 함유하는 바이오필름 형성 억제용 조성물
Nasser Anti-bactericidal and anti-biofilm activities of silver nanoparticles against multidrug-resistant Gram-negative bacilli isolated from burn wound infections
CN103860589A (zh) 一类具有抗细菌和真菌作用的稀土化合物

Legal Events

Date Code Title Description
NENP Non-entry into the national phase

Ref country code: DE

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17795261

Country of ref document: EP

Kind code of ref document: A1