CN110251499A - Dandelion extract and its purposes in preparation prevention and treatment woman vagina disease medicament - Google Patents
Dandelion extract and its purposes in preparation prevention and treatment woman vagina disease medicament Download PDFInfo
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- CN110251499A CN110251499A CN201910498473.1A CN201910498473A CN110251499A CN 110251499 A CN110251499 A CN 110251499A CN 201910498473 A CN201910498473 A CN 201910498473A CN 110251499 A CN110251499 A CN 110251499A
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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
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid
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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
- A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
- A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
- A61K31/216—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acids having aromatic rings, e.g. benactizyne, clofibrate
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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
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
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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
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
- A61K31/7056—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing five-membered rings with nitrogen as a ring hetero atom
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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)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/288—Taraxacum (dandelion)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/02—Drugs for genital or sexual disorders; Contraceptives for disorders of the vagina
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- 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
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- 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
- A61P31/10—Antimycotics
-
- 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
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- 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
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- 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
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- 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
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
-
- 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
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
-
- 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
Abstract
The purposes in woman vagina disease medicament is prevented and treated the invention discloses dandelion extract and its in preparation, the dandelion extract is mainly by 4- coumaric acid, ferulic acid, Quercetin pentoside, 3, bis--O- caffeoyl guinic acid of 5-, 4,5-, bis--O- caffeoylquinic acids and luteolin composition.Test result is shown, dandelion extract dialogue candidiasis and Gardnerella have good inhibiting effect, the minimum inhibitory concentration of dialogue candidiasis and Gardnerella is respectively 0.4mg/mL and 0.2mg/mL, can be used for preparing prevention and treatment by candida albicans bacterium and Gardnerella and causes woman vagina disease medicament.The raw material drawing of dandelion extract used in the present invention is convenient, and production cost is low, good product quality, cheap, environmentally protective, is suitble to promote and apply.Therefore, a kind of novel Chinese medicine vaginopathy therapeutic agent can be become.
Description
Technical field
The invention belongs to technical field of pharmaceuticals, and in particular to a kind of dandelion extract and the extract are pre- in preparation
Purposes anti-and that treatment is in the woman vagina disease medicament as caused by candida albicans bacterium and Gardnerella.
Background technique
Vaginopathy also known as colpitis are the illnesss that Out-patient Clinic of Department of Gynecology is common and frequently occurs.The healthy vagina of women is to thin
The invasion of the pathogenic microorganisms such as bacterium, fungi has natural protection ability, and when the natural defending system of healthy vagina is destroyed, disease
Substance is availed oneself of the opportunity to get in, and original ecological balance between flora and vagina, the growth conditions that certain pathogenic bacteria are suitable for herein have been broken
Under mushroom out, and then cause colpitis.
Due to diagnosed in clinical position it is incorrect, treatment specification is understood it is not thorough and to some new vagina infections, mixed
Closing womankind in infection understanding deficiency and daily life, correctly understanding, shortage are relevant certainly comprehensively to gynecological disease shortage
Body health care, a variety of bad daily life systems etc., make physiological health go from bad to worse in addition, and some women is caused to be possessed by the demon of disease throughout the year, and
Recurrent exerbation or obstinate bring inconvenience to personal normal work and study, family life.
Clinically common vaginopathy has: bacterial vaginosis BV, candidiasis vaginosis, trichomonad vaginosis, old
Year vaginosis and youngest daughter's vaginosis.Wherein bacterial vaginosis BV and candidiasis vaginosis are two kinds of common property vaginas
Disease, they are mainly microbial by Gardnerella and candida albicans respectively.In recent years, the abuse of antibiotic causes bacterium
With many bio-safety problems such as antifungal agent resistance, so finding a kind of good natural medicinal ingredients of bacteriostasis property as vaginopathy
Therapeutic agent is particularly important.
Summary of the invention
The object of the present invention is to provide a kind of dandelion extracts, and provide a kind of new use for the dandelion extract
On the way.
For above-mentioned purpose, dandelion extract of the invention includes the ingredient of following relative mass percentage compositions: 4- is fragrant
Beans acid 11%~12%, ferulic acid 3.5%~4.5%, Quercetin pentoside 10%~11%, bis--O- caffeoyl Kui Buddhist nun of 3,5-
Sour 34%~35%, bis--O- caffeoylquinic acids 3.5%~4.5% of 4,5-, luteolin 11%~12%.
Above-mentioned dandelion extract is obtained by following methods:
(1) water mentions
It takes dry dandelion herb to crush and crosses 60~80 meshes, distilled water is added according to solid-liquid ratio 1g:25~35mL,
30~40min is extracted at 50~70 DEG C, is extracted 2 times, is filtered, merging filtrate, and filtrate is concentrated 2~3 times, is centrifugated, and collects
Supernatant.
(2) preliminary purification
5~10min of ultrasound after the filter membrane in 0.45 μm of aperture of supernatant is filtered, the D101 macropore then above pre-processed
Adsorption resin column after washing 2 column volumes, is eluted with the ethanol water that volumetric concentration is 50%~70%, collects eluent,
Eluent is concentrated 4~6 times.
(3) it purifies again
The isometric ethyl acetate of eluent after concentration is extracted, after collecting extract liquor and being concentrated 5~6 times, vacuum
It is dry, obtain dandelion extract.
Dandelion extract of the present invention woman as caused by candida albicans bacterium and Gardnerella in preparation prevention and/or treatment
Purposes in cysthus tract disease drug.Test result shows, dandelion extract dialogue candidiasis of the present invention and Gardner
Bacterium has a good inhibiting effect, the minimum inhibitory concentration of dialogue candidiasis and Gardnerella be respectively 0.4mg/mL and
0.2mg/mL can be used for preparing prevention and treatment by candida albicans bacterium and Gardnerella and cause woman vagina disease medicament.
The raw material drawing of dandelion extract used in the present invention is convenient, and production cost is low, good product quality, and price is low
It is honest and clean, it is environmentally protective, it is suitble to promote and apply.Therefore, a kind of vaginopathy therapeutic agent of novel pure Chinese medicine can be become.
Detailed description of the invention
Fig. 1 is the HPLC analysis chart for the dandelion extract that embodiment 1 obtains, wherein 1 is 4- coumaric acid；2 be ferulic acid；
3 be Quercetin pentoside；4 be bis--O- caffeoyl guinic acid of 3,5-；5 be bis--O- caffeoylquinic acids of 4,5-；6 be reseda
Element.
Fig. 2 is the bacteriostatic test for the dandelion extract dialogue candidiasis that embodiment 1 obtains.
Fig. 3 is bacteriostatic test of the obtained dandelion extract of embodiment 1 to Gardnerella.
Specific embodiment
Invention is further described in detail with reference to the accompanying drawings and examples, but protection scope of the present invention not only limits
In these embodiments.
Embodiment 1
Prepare dandelion extract
(1) water mentions
It takes fresh dandelion herb to dry to constant weight in 60 DEG C of vacuum, crushed after drying and crosses 80 meshes, then according to feed liquid
Distilled water is added than 1g:30mL, 30min is extracted at 60 DEG C, extracts 2 times, filtering, merging filtrate, filtrate is concentrated into 2~3 times,
It is centrifuged 10min in 8000r/min, collects supernatant.
(2) preliminary purification
After the filter membrane nutsch filter suction filtration in 0.45 μm of aperture of the supernatant that step (1) is collected, ultrasonic 10min, then on
The D101 large pore resin absorption column pre-processed after washing 2 column volumes, is carried out with the ethanol water that volumetric concentration is 60%
Eluent is collected in elution, and eluent is concentrated 4~6 times.
(3) it purifies again
The isometric ethyl acetate of eluent after step (2) concentration is extracted, extract liquor is collected and is concentrated 5~6 times
Afterwards, it is dried in 60 DEG C of vacuum ovens, obtains dandelion extract.
Using high performance liquid chromatography (HPLC) measure gained dandelion extract composition, HPLC system by one equipped with
The Agilent1100HPLC system and diode array detector of DiKMADiamonsil C18 column (250mm × 4.6mm, 5 μm)
(DAD) it forms.The methanol aqueous solution that sample volumetric concentration is 80% dissolves, and column temperature is maintained at 40 DEG C, and mobile phase (is washed by acetonitrile
De- liquid A) and volumetric concentration be 0.1% phosphate aqueous solution (eluent B) form, the flow velocity of isocratic elution liquid is 1.0mL/min,
47min is eluted according to 20%A, 80%B, sample volume is 10 μ L.The peak area of all polyphenolic substances is surveyed at 280nm wavelength
It is fixed.As shown in Figure 1, HPLC detects 6 kinds of compounds, it is 4- coumaric acid, ferulic acid, Quercetin pentoside, 3,5-, bis--O- respectively
Caffeoyl guinic acid, 4,5-, bis--O- caffeoylquinic acids and luteolin, this 6 kinds of compounds percentage in this extract
Content is respectively 11.45%, 3.96%, 10.48%, 34.24%, 3.91% and 11.80%.
Embodiment 2
The dandelion extract dialogue candidiasis of embodiment 1 and the rejection of Gardnerella
(1) bacteriostatic activity is tested
Dandelion extract dialogue candidiasis (Candida albicans, CMCC85021) is measured by paper disk method
With the bacteriostasis of Gardnerella (Gardnerella vaginalis, ATCC14018).Experimental strain is recovered, is activated 2 times.
The passage of candida albicans bacterium cultivates 12h in husky formula fluid nutrient medium, in 37 DEG C, 190r/min；Gardnerella is trained using HTM agar
Base plate is supported in 37 DEG C of 5%CO2Under the conditions of cultivate 12h, bacterium colony is washed to get off to be prepared into bacteria suspension with fluid nutrient medium.With point
The respective thallus OD of light photometric determination600nmValue.Work as OD600nmIt is best coating concentration when=0.6~0.7；If OD600nm>0.7
When, it is suitably diluted with fluid nutrient medium, then be coated.Husky formula agar medium and HTM agar medium are prepared, respectively
Sterilize 20min at 115 DEG C and 121.3 DEG C, and sterile cozymase (NAD) solution is cooled to 50 DEG C of addition HTM agar trainings in culture medium
Base is supported, mixes, prepares plate respectively.The bacteria suspension that activated and concentration is suitable for 100 μ L of liquid-transfering gun absorption is used in planar surface
Sterile L shape glass rod smoothens bacterium solution.Experimental group: 10 μ L are drawn with the sample solution of 40% dmso solution and are added drop-wise to diameter
6mm aseptic filter paper on piece, appropriateness are toasted, and planar surface is affixed on.Blank group: sterile 40% dimethyl sulfoxide (DMSO) of 10 μ L is dripped
It is added to diameter 6mm aseptic filter paper on piece, appropriateness is toasted, and planar surface is affixed on.Respectively at 37 DEG C, 5%CO2Under the conditions of be inverted culture
18~36h, observation experiment is as a result, measurement antibacterial circle diameter size.
Test result is shown, shows dimethyl sulfoxide to fungi without inhibition zone substantially around the blank group 40%DMSO scraps of paper
With bacterial growth without influence；There is inhibition zone around experimental group dandelion extract, shows dandelion extract to candida albicans
Bacterium and Gardnerella have obvious bacteriostasis (see Fig. 2).When dandelion extract concentration be 0.40~2.28mg/mL, it is white
The antibacterial circle diameter of candidiasis is 6.00~10.53mm, when wherein dandelion extract concentration is 2.28mg/mL, dialogue
The average antibacterial circle diameter of candidiasis is 10.38 ± 0.15mm；When the concentration of dandelion extract is 0.20~3.80mg/
ML, the antibacterial circle diameter of Gardnerella is 6.00~11.85mm, when wherein dandelion extract concentration is 3.80mg/mL, dialogue
The average antibacterial circle diameter of candidiasis is 11.86 ± 0.17mm.Illustrate dandelion extract dialogue candidiasis and adds
Moral receives bacterium with good inhibitory effect.
(2) minimum inhibitory concentration
Candida albicans bacterium and Gardnerella are cultivated according to method (1), is transferred to bacterial concentration with fluid nutrient medium
107CFU/mL, and prepare 4mg/mL dandelion extract mother liquor.Pu is added into the test tube equipped with 2mL fluid nutrient medium respectively
Public English extract mother liquor, make in culture medium dandelion extract ultimate density 0,0.05,0.10,0.20,0.40,0.60,
0.80mg/mL is 10 by 4% (v/v) access concentration7The bacterium solution of CFU/mL, Gardnerella test tube use paraffin vaseline
(VParaffin oil:VVaseline=1:1 mixing) seal liquid level.Candida albicans bacterium and Gardnerella are respectively at 37 DEG C, 190r/min and 37 DEG C
It cultivates for 24 hours, as sample OD600nmThe concentration for changing the dandelion extract less than 5% for the first time is minimum inhibitory concentration, as MIC.
It is found through analysis, when the concentration of dandelion extract is greater than 0.4mg/mL, candida albicans bacterium OD600nmValue variation is less than for the first time
5%, when the concentration of dandelion extract is greater than 0.2mg/mL, Gardnerella OD600nmValue variation is for the first time less than 5%, it is seen then that Pu
The MIC value of public English extract dialogue candidiasis and Gardnerella is respectively 0.4mg/mL and 0.2mg/mL.
From the foregoing, it can be seen that dandelion extract dialogue candidiasis of the present invention and Gardnerella are imitated with good inhibition
The MIC value of fruit, dialogue candidiasis and Gardnerella is respectively 0.4mg/mL and 0.2mg/mL, can be used for preparing prevention and/
Or treatment woman vagina disease medicament as caused by candida albicans bacterium and Gardnerella.
Claims (3)
1. a kind of dandelion extract, it is characterised in that: the dandelion extract includes following relative mass percentage compositions
Ingredient: 4- coumaric acid 11%~12%, ferulic acid 3.5%~4.5%, Quercetin pentoside 10%~11%, bis--O- of 3,5-
Caffeoyl guinic acid 34%~35%, bis--O- caffeoylquinic acids 3.5%~4.5% of 4,5-, luteolin 11%~12%.
2. dandelion extract according to claim 1, it is characterised in that the dandelion extract is by following methods
It obtains:
(1) water mentions
It takes dry dandelion herb to crush and crosses 60~80 meshes, distilled water is added according to solid-liquid ratio 1g:25~35mL, in 50
30~40min is extracted at~70 DEG C, is extracted 2 times, is filtered, merging filtrate, and filtrate is concentrated 2~3 times, and supernatant is collected in centrifuge separation
Liquid；
(2) preliminary purification
5~10min of ultrasound after the filter membrane in 0.45 μm of aperture of supernatant is filtered, the D101 macroporous absorption then above pre-processed
Resin column after washing 2 column volumes, is eluted with the ethanol water that volumetric concentration is 50%~70%, collects eluent, will wash
De- liquid is concentrated 4~6 times；
(3) it purifies again
The isometric ethyl acetate of eluent after concentration is extracted, after collecting extract liquor and being concentrated 5~6 times, vacuum drying,
Obtain dandelion extract.
3. dandelion extract described in claim 1 is in preparation prevention and/or treats by candida albicans bacterium and Gardnerella
Purposes in caused woman vagina disease medicament.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113577123A (en) * | 2021-08-23 | 2021-11-02 | 江苏海洋大学 | Preparation method and application of dandelion extract |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113577123A (en) * | 2021-08-23 | 2021-11-02 | 江苏海洋大学 | Preparation method and application of dandelion extract |
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