CN102988456A - Application of honeysuckle extractive from active components of Qingkailing for preparing multiple-resistant bacteria resistant medicine - Google Patents
Application of honeysuckle extractive from active components of Qingkailing for preparing multiple-resistant bacteria resistant medicine Download PDFInfo
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Classifications
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- 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
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- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a honeysuckle extractive from active components of Qingkailing, which has inbibitional effects on Acinetobacter calcoaceticus, Acinetobacter baumannii, A.baumannii, Stenotrophomonasmaltophilia and Escherichia coli containing NDM-1 drug resistant genes, therefore, the honeysuckle extractive can be used for inhibiting the bacterial diseases and curing and/or preventing diseases caused by bacteria.
Description
Technical field
The present invention relates to a kind of new medical usage of Flos Lonicerae, specifically prepare application in the anti-multi-drug resistant bacteria medicine at Flos Lonicerae extract.
Background technology
Multi-drug resistant bacteria (multiple resistant bacteria) refers to the pathogen of multi-drug resistant, i.e. a kind of microorganism is to three classes (such as aminoglycoside, erythromycin, beta-lactam class) or drug resistance of above antibiotic while of three classes.Wherein the most common is Acinetobacter bauamnnii and the Pseudomonas aeruginosa that occurs among NDM-1, MDR-TB, MDR-MRSA and the ICU that is everlasting.NDM-1(New Delhi metallo-β-lactamase-1, the New Delhi metallo-β-lactamase), have another name called the NDM-1 antibacterial, be a kind of enzyme of Carbapenem-resistant class.The NDM-1 gene is often carried by plasmid, but also is incorporated on the chromosome of antibacterial, is easy to propagate between different bacterial strains and diffusion, and the protein monomer molecular weight of NDM-1 coding is 28kDa, and the sequence similarity of its sequence and other MBL is very low.NDM-1 mainly is distributed in enterobacteriaceae and the Acinetobacter bauamnnii, and the antibacterial of carrying NDM-1 not only comprises conditioned pathogen, also comprises pathogenic bacterium, such as common shigella, cholera, pseudomonas aeruginosa etc.It has the height drug resistance, almost can be hydrolyzed clinical line antibacterials commonly used, comprises beta-lactam, carbapenems, fluoroquinolones, aminoglycoside etc.For a long time, the abuse of antibiotic medicine is the major reason that expedites the emergence of multi-drug resistant bacteria.At present, multi-drug resistant bacteria infects and presents growth trend, but the Antimicrobial drug choice scheme seldom, and therefore heavy drug-fast bacteria infection has become global thorny problem.
Flos Lonicerae (dry flower of caprifoliaceae plant Radix Ophiopogonis Lonicera japonica Thunb., Flos Lonicerae Lonicera hypoglauca Miq., Flos Lonicerae Lonicera confusa DC. or hair style Radix Ophiopogonis Lonicera dasystyla Rehd. or the flower that band is just opened) is the primary raw material medicine of heat-clearing and toxic substances removing herbal mixture QINKAILING ZHUSHEYE, effect with dispelling wind-heat, removing summer-heat blood poison, to various febrile diseases, has remarkable therapeutic effect such as cards such as fever of the body, dermexanthesis, a speckle, pyretic toxicity carbuncle sore, laryngopharynx swelling and pain.The how better medical efficacy of R and D Flos Lonicerae is one of the important subject in many Chinese medicine and pharmacies field.
Summary of the invention
Purpose of the present invention is exactly to develop the novel medical use of Flos Lonicerae extract, provides a kind of effectively medicine of antagonism multi-drug resistant bacteria for clinical simultaneously.
The present inventor finds that through studying for a long period of time Flos Lonicerae extract has significantly inhibitory action to containing NDM-1 drug resistant gene antibacterial, and be directly proportional with drug level, thereby invented a kind of new medical usage of Flos Lonicerae extract, namely finished the application of Flos Lonicerae extract in the anti-multi-drug resistant bacteria medicine of preparation.
Flos Lonicerae extract of the present invention can carry out according to the Traditional Chinese medicine extraction method of routine.
The present invention is as follows in the method that this provides:
(1) extracting honeysuckle is an amount of, adds water boil twice, filters, and merges filtrate twice;
(2) gained filtrate is concentrated into 1～2 times of amount of crude drug volume, adds ethanol, and cold preservation is filtered, and filtrate recycling ethanol is nothing alcohol flavor extremely, thin up, and regulating pH value is to faintly acid, filtration;
(3) with the upper macroporous adsorbent resin of gained filtrate, water, ethanol elution are collected eluent successively, filter, and decompression recycling ethanol gets Flos Lonicerae extract, cold preservation to without the alcohol flavor.
Wherein, step adds water boil in (1), adds 10 times of amounts of water for the first time, boils 1 hour, adds 8 times of amounts of water for the second time, boils 0.5 hour; Add ethanol in the step (2) and make and contain alcohol amount and reach 80%, cold preservation 2～4 days, thin up to relative density is 1.05～1.2(60 ℃), described pH value is 5; In the step (3), the consumption of eluant water is 6～8 times of amount column volumes, and ethanol is 20% ethanol of 10～12 times of amount column volumes.
Flos Lonicerae extract described in the present invention can be prepared into according to the described method of prior art the pharmaceutical preparatioies such as Flos Lonicerae granule, Flos Lonicerae electuary, Flos Lonicerae injection, Flos Lonicerae capsule.
Multi-drug resistant bacteria described in the present invention refers to contain the multi-drug resistant bacteria of NMD-1 gene, comprise acinetobacter calcoaceticus (Acinetobacter calcoaceticus), Acinetobacter bauamnnii (Acinetobacter baumannii, A.baumannii), have a liking for the narrow food Zymomonas mobilis of Fructus Hordei Germinatus oligotrophy (Stenotrophomonas maltophilia) or contain NDM-1 drug resistant gene escherichia coli (Escherichia coli).
It is gram negative bacteria that the above-mentioned NDM-1 of containing drug resistant gene detects bacterial strain, has stronger pathogenic and antibiotic resistance.Wherein, acinetobacter calcoaceticus and Acinetobacter bauamnnii can cause pulmonary infection, wound and skin infection, urogenital infections, bacteremia bacteremia, meningitis etc., the narrow food Zymomonas mobilis of Fructus Hordei Germinatus oligotrophy can cause serious respiratory system infection etc., contains NDM-1 drug resistant gene escherichia coli and can cause to intestinal infection and to antibiotic and have Drug resistance.
Further, the present invention also provides the application of Flos Lonicerae extract in the medicine of the anti-acinetobacter calcoaceticus of preparation, and namely Flos Lonicerae extract treats and/or prevents application in the medicine of the disease that acinetobacter calcoaceticus causes in preparation.
Further, the present invention also provides the application of Flos Lonicerae extract in the medicine of the anti-Acinetobacter bauamnnii of preparation, and namely Flos Lonicerae extract treats and/or prevents application in the medicine of the disease that Acinetobacter bauamnnii causes in preparation.
Further, the present invention also provides the application of Flos Lonicerae extract in the anti-medicine of having a liking for the narrow food Zymomonas mobilis of Fructus Hordei Germinatus oligotrophy of preparation, and namely Flos Lonicerae extract treats and/or prevents application in the medicine of having a liking for the microbial disease of the narrow food unit cell of Fructus Hordei Germinatus oligotrophy in preparation.
Further, the present invention also provides Flos Lonicerae extract preparing the application that resists in the medicine that contains NDM-1 drug resistant gene antibacterial.
Further, the present invention also provides Flos Lonicerae extract in the anti-application that contains in the colibacillary medicine of NDM-1 drug resistant gene of preparation, and namely Flos Lonicerae extract treats and/or prevents application in the medicine that contains the disease that NDM-1 drug resistant gene escherichia coli cause in preparation.
Flos Lonicerae preparation of the present invention can be used for treating above-mentioned disease by containing the initiation of NDM-1 drug resistant gene multidrug resistant antibacterial, as contain intestinal infection, respiratory system infection, wound and skin infection that NDM-1 drug resistant gene multidrug resistant antibacterial causes, urogenital infections, bacteremia bacteremia, meningitis etc., especially be applicable to treat by acinetobacter calcoaceticus, Acinetobacter bauamnnii, have a liking for the narrow food Zymomonas mobilis of Fructus Hordei Germinatus oligotrophy or contain the disease that NDM-1 drug resistant gene escherichia coli cause.
Pharmaceutical preparation of the present invention can be determined according to clinical experiment and patient's concrete condition during its concrete consumption when using as purposes of the present invention.
Specific embodiments
Further set forth by the following examples the beneficial effect of medicine of the present invention with test example.
Embodiment 1
The preparation method of Flos Lonicerae extract
(1) extracting honeysuckle is an amount of, adds water boil twice, adds the water of 10 times of amounts for the first time, boils 1 hour, adds the water of 8 times of amounts for the second time, boils 0.5 hour, filters, and merges filtrate twice; When (2) gained filtrate is concentrated into 1～2 times of crude drug amount volume, adds ethanol and make and contain the alcohol amount and reach 80%, cold preservation 2～4 days is filtered, and filtrate recycling ethanol is to without the alcohol flavor, and thin up is 1.05～1.2(60 ℃ to relative density), regulating pH value is 5, filtration; (3) with the upper macroporous adsorbent resin of gained filtrate, with the water elution of 6～8 times of amount column volumes, 20% ethanol elution of 10～12 times of amount column volumes, collect eluent successively, filter, decompression recycling ethanol gets Flos Lonicerae extract, cold preservation to without the alcohol flavor.
Pharmacodynamics test
Test example 1
Flos Lonicerae extract is to containing the fungistatic effect research of NDM-1 drug resistant gene antibacterial
The detection bacterial strain that this pharmacodynamics test adopts: acinetobacter calcoaceticus (Acinetobacter calcoaceticus, A.calcoaceticus), Acinetobacter bauamnnii (Acinetobacter baumannii, A.baumannii) and have a liking for narrow food Zymomonas mobilis (the Stenotrophomonas maltophilia of Fructus Hordei Germinatus oligotrophy, S.maltophilia), all contain the blaNDM-1 drug resistant gene; PGEX-4T-NDM1-DH5 α is the escherichia coli (Escherichia coli, E.coli) that contain the NDM-1 recombiant plasmid; PGEX-4T-NDM-1-BL21(expresses NDM-1 in e. coli bl21) be the escherichia coli (Escherichia coli, E.coli) that contain the NDM-1 recombiant plasmid, on its plasmid with the GST label.Above-mentioned bacterial strains provides by Diseases Preventing and Controlling Institute.
Preparation LB agar culture medium, when high pressure was cooled to 60 ℃, the extracting honeysuckle extract was pressed the 1:9(medicine: LB, V/V) ratio preparation variable concentrations medicine (0.1g/ml, 0.2g/ml, 0.3g/ml) flat board, agar thickness 3-4mm, the preparation medicine is dull and stereotyped, for subsequent use.
The research Flos Lonicerae carries out bacteriostatic experiment to containing NDM-1 drug resistant gene antibacterial:
4 strains are contained NDM-1 gene Resistant strain acinetobacter calcoaceticus, Acinetobacter bauamnnii, have a liking for the narrow food Zymomonas mobilis of Fructus Hordei Germinatus oligotrophy and escherichia coli pGEX-4T-NDM1-DH5 α is inoculated in respectively in the LB fluid medium that contains 16 μ g/ml imipenums, 37 ℃, 200rpm overnight incubation, after next day, bacterium liquid diluted 10 times with aseptic PBS buffer, drawing respectively 2 μ l is inoculated in the medicine agar plate surface or adopts the method for scoring inoculation, flat board places 37 ℃ to hatch about 24h, observe each strain growth situation, get the biocidal property measurement result of Flos Lonicerae extract, see Table 1.
Adopt agar dilution that the minimal inhibitory concentration (MIC) of Flos Lonicerae extract is measured, Resistant strain is inoculated on the agar plate of different pharmaceutical concentration, hatch about 24h for 37 ℃, observe each strain growth situation, the minimum drug level (MIC value) that the inhibition bacterial strain can not be grown is respectively acinetobacter calcoaceticus 0.2g/ml, Acinetobacter bauamnnii 0.2g/ml, has a liking for the narrow food Zymomonas mobilis of Fructus Hordei Germinatus oligotrophy 0.1g/ml, pGEX-4T-NDM-1-BL21 0.2g/ml.
Experimental result shows: Flos Lonicerae all has inhibitory action to Resistant strain, and along with the increase of drug level, inhibitory action obviously strengthens, and illustrate that Flos Lonicerae has inhibitory action to containing NDM-1 drug resistant gene fastbacteria, and fungistatic effect is obvious; Medicine is different to the antibacterial ability that difference detects bacterial strain, and MIC is difference to some extent also, has the anti-ability that contains NDM-1 drug resistant gene fastbacteria of stronger wide spectrum.
Table 1 Flos Lonicerae extract concentration is on the impact of bacterial strain
Annotate: +++expression does not suppress growth, ++ expression suppresses not obvious, and+expression obviously suppresses, and-expression suppresses fully, does not grow.
Claims (7)
1. the application of Flos Lonicerae extract in the medicine of the anti-multi-drug resistant bacteria of preparation.
2. Flos Lonicerae extract is preparing the application that resists in the medicine that contains NDM-1 drug resistant gene antibacterial.
3. the application of Flos Lonicerae extract in the medicine of the anti-acinetobacter calcoaceticus of preparation.
4. the application of Flos Lonicerae extract in the medicine of the anti-Acinetobacter bauamnnii of preparation.
5. Flos Lonicerae extract is preparing the application that resists in the medicine of having a liking for the narrow food Zymomonas mobilis of Fructus Hordei Germinatus oligotrophy.
6. Flos Lonicerae extract is in the anti-application that contains in the colibacillary medicine of NDM-1 drug resistant gene of preparation.
7. the described application of any one according to claim 1-6, it is characterized in that, described Flos Lonicerae extract is to add water boil twice by Chinese medicine honeysuckle, filter, merge filtrate twice, add ethanol after gained filtrate is concentrated and make and contain the alcohol amount and reach 80%, cold preservation, filter, filtrate recycling ethanol is to distinguishing the flavor of without alcohol, and thin up is 1.05～1.2(60 ℃ to relative density), regulating pH value is 5, filter, with the upper macroporous adsorbent resin of gained filtrate, successively with 6～8 times of water elutions of measuring column volumes, 20% ethanol elution of 10～12 times of amount column volumes, collect eluent, filter, decompression recycling ethanol to nothing alcohol is distinguished the flavor of, and get final product.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105878470A (en) * | 2016-06-19 | 2016-08-24 | 神威药业集团有限公司 | Qingkailing pharmaceutical composition |
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| CN102114216A (en) * | 2010-12-03 | 2011-07-06 | 郑延明 | Externally applied antibacterial and antiphlogistic medicament and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105878470A (en) * | 2016-06-19 | 2016-08-24 | 神威药业集团有限公司 | Qingkailing pharmaceutical composition |
| CN105878470B (en) * | 2016-06-19 | 2019-07-05 | 神威药业集团有限公司 | A kind of Qing kailing pharmaceutical composition |
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