CN109394796A - Lactobacillus crispatus preparation and its application - Google Patents
Lactobacillus crispatus preparation and its application Download PDFInfo
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- CN109394796A CN109394796A CN201811488762.5A CN201811488762A CN109394796A CN 109394796 A CN109394796 A CN 109394796A CN 201811488762 A CN201811488762 A CN 201811488762A CN 109394796 A CN109394796 A CN 109394796A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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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
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The present invention relates to a kind of probiotics and its applications, and in particular to main active includes that the probiotics of Lactobacillus crispatus and said preparation are preventing or treating the application in related disease.
Description
Technical field
The present invention relates to a kind of probiotics and its applications, and in particular to main active includes Lactobacillus crispatus
Probiotics and said preparation are preventing or are treating the application in related disease.
Background technique
Lactobacillus crispatus is the common beneficial bacterium of vagina, has played important function for maintenance vagina health, but
Due to abuse of antibiotics etc., vagina Lactobacillus crispatus is caused to largely reduce, large intestine bar occurs for vaginal dysbacteriosis
Bacterium, staphylococcus aureus or candida albicans infection cause various vaginal diseases to occur.It separates Lactobacillus crispatus and develops micro- life
State preparation becomes the hot spot studied at present, but the screening for the Lactobacillus crispatus that growth activity is high, inhibition pathogenic bacteria effect is good is mesh
Preceding difficult point.
The present inventor has found that Lactobacillus crispatus DH200 has high bioactivity after study, and main active includes
The probiotics of Lactobacillus crispatus DH200 can effectively adjust body microecological balance, prevention or treatment related disease, safety
There is no any toxic side effect, there is huge economic value and social value, and has no that correlative study is reported, spy's application the invention
Patent.
Summary of the invention
The object of the present invention is to provide a kind of probiotics, the main active of said preparation includes DH200, and preservation is compiled
Number CGMCC No.16193, and said preparation is provided and is preventing or treating the application in related disease.
Confirm that bacterial strain exists according to results such as bacterium cell form, physiological and biochemical property, 16S rRNA sequence comparative analysis
Taxonomy, the bacterial strain of discovery number DH200 are Lactobacillus crispatus；Feature of this plant of bacterium on morphology and general aspects is such as
Under:
Cellular morphology and physical and chemical experiment result:
Lactobacillus crispatus DH200 of the present invention inhibits the work of Escherichia coli, staphylococcus aureus or Candida albicans
With significant, have and prevent or the application for the treatment of Escherichia coli, staphylococcus aureus or candida albicans infection, have prevention or
Treat the application of vaginitis, vaginitis includes bacterial vaginosis BV, mycotic, senile vahinitis, and colpomycosis is into one
Step includes monilial vaginitis.
Probiotics of the invention, which also can be used as effect components and apply, to be used in compositions related prevent or control
Treat above-mentioned disease.
The present inventor is repeatedly screened and is cultivated, and isolated one plant of Lactobacillus crispatus bacterial strain DH200, the bacterial strain is compared with other
Bacterial strain has higher activity, superior adjusting body microecological balance ability and prebiotic effect.Curling cream of the present invention
Bacillus further includes the bacterial strain derived from Lactobacillus crispatus DH200, especially has high activity, or micro- with excellent adjusting body
The derivative strain of ecological balance ability and prebiotic effect.Main active includes Lactobacillus crispatus DH200 and its derivative strain
Probiotics each fall within protection scope of the present invention.
Probiotics of the present invention, main active include but is not limited to Lactobacillus crispatus DH200, deposit number
The oligosaccharide such as other viable bacterias, stachyose, oligofructose or other active components can also be added in CGMCC No.16193.
Probiotics of the invention are it using the Lactobacillus crispatus as main active or main active
One, according to certain preparation process, conventional excipient is added, flavoring agent, disintegrating agent, preservative, lubricant, wetting agent, sticks
The auxiliary materials such as mixture, solvent, thickener, solubilizer, the dosage form that manufactured any one is adapted for use with, such as tablet, suppository, capsule
The dosage forms such as agent, granule, powder, liquid preparation, pulvis.
Probiotics of the present invention include but is not limited to composite medicine, health care product, food or sterile products, disinfection
Product further comprises inhibiting-bacteria preparation.
Lactobacillus crispatus described in this probiotics refers to bion living.
Lactobacillus crispatus described in this probiotics has high bioactivity.
This probiotics, the total viable count of the Lactobacillus crispatus that solid pharmaceutical preparation includes are not less than 1 × 106CFU/g generally exists
1×107CFU/g or more can reach 1 × 1012CFU/g or 1 × 1012CFU/g or more；Or the curling cream that liquid preparation includes
The total viable count of bacillus is not less than 1 × 106CFU/mL, generally 1 × 107CFU/mL or more can reach 1 × 1012CFU/mL
Or 1 × 1012CFU/mL or more.
Specific embodiment
Preparation example explanation: it is above-mentioned that the preparation of Lactobacillus crispatus preparation is illustrated, pass through Lactobacillus crispatus here
It is specifically described for DH200, preparation method those skilled in the art of other Lactobacillus crispatus strain preparations pass through this reality
It applies example to be easy to grasp, preparation method those skilled in the art of other dosage forms are easy to grasp by this implementation, herein no longer
Explanation is described one by one.Preparation method is not limited to described in the embodiment of the present invention, the well known method that can reach preparation purpose
It can be with it is not limiting the scope of the invention that the preparation explanation of embodiment, which is the description of the invention,.
Prepare the preparation of 1 Lactobacillus crispatus pulvis of embodiment
The preparation of 1 bacterium powder and the identification of strain
The samples such as vagina secretion are taken, then sample is placed in sterilizing bottle, therefrom take 2 grams of samples that 18mL is added
It in the dilution of sterilizing, mixes well, in aseptic operating platform, carries out 10-1、10-2、10-3、 10-4, 10-5, 10-6, 10-7Gradient
Dilution, takes 10-5, 10-6, 10-7Three dilution gradients, are coated on MRS solid medium, are placed in incubator, cultivate at 37 DEG C
48 hours, the single colonie to grow fine is selected to be inoculated into MRS fluid nutrient medium, amplification cultivation 48 hours at 37 DEG C.By gained
After medium centrifugal (12000rpm) isolates thallus, by thallus vacuum freezedrying, dry bacterium powder is modulated, viable count is equal
It is 1 × 1010Then CFU/g or more carries out strain idenfication.Volume is accredited as through Physiology and biochemistry and 16S rRNA sequence comparative analysis
Bent lactobacillus is Lactobacillus crispatus DH200, deposit number CGMCC No.16193.
2 toxicity tests
2.1 animals and grouping take 20 SPF rank mouse, 6-8 week old, and weight 14-18g is randomly divided into Lactobacillus crispatus
Group and non-administered group, every group 10.
Above-mentioned Lactobacillus crispatus bacterium powder is modulated to containing bacterium number be 1 × 10 by 2.2 preparation bacterium solutions with purified water9CFU/mL
Bacterium solution.
2.3 method Lactobacillus crispatus groups and non-administered group give identical basal feed, and rearing conditions are consistent, volume
Bent lactobacillus group gavages Lactobacillus crispatus bacterium solution 0.5mL daily, and non-administered group gavages purified water 0.5mL daily, feeds 14 days,
Observe weight and toxic reaction.
2.4 result
Each group mouse does not occur abnormal conditions, and chatter, spasm, ataxia, posture exception do not occur, and no eyeball is prominent
Out, urination is normal, and skin, breathing are normal, and no death condition has no toxic reaction.
3 are prepared into tablets and other formulations
It is separated and identified according to above-mentioned steps and method, through experimental check nontoxicity, so that it may by Lactobacillus crispatus strain system
At bacterium powder, then it is added as required relevant auxiliary materials various dosage forms is made and pressed preferably according to the viable count of Lactobacillus crispatus bacterium powder
Tablet is made in ratio addition microcrystalline cellulose, sodium carboxymethyl starch, magnesium stearate etc., makes viable count not less than 1 × 107CFU/g,
Then it bottles.
The present inventor uses above-mentioned separation and preparation method, while also separation obtains Lactobacillus crispatus DH144 and curling
Lactobacillus DH134.
Application effect embodiment explanation:
For the present invention with Lactobacillus crispatus DH200, probiotics made of deposit number CGMCC No.16193 are representative
Illustrate the application effect of Lactobacillus crispatus preparation.It should be understood that these embodiments are merely to illustrate the present invention rather than limitation originally
The range of invention.To better illustrate application effect of the invention, the present inventor also uses Lactobacillus crispatus DH144, curling cream
Probiotics made of bacillus DH134 and commercially available Lactobacillus crispatus compare explanation.Commercially available volume used in the present invention
Active polypeptide lactobacillus product of the bent lactobacillus from market sale.Combined with specific embodiments below, this hair is further described
It is bright.
Application effect embodiment 1: the bioactivity of Lactobacillus crispatus
The preparation of 1 bacterium powder has been prepared Lactobacillus crispatus DH200, has been crimped newborn bar using the preparation method of above-mentioned bacterium powder
Bacterium DH144, Lactobacillus crispatus DH134 and commercially available Lactobacillus crispatus bacterium powder.
It is 1 × 10 that above-mentioned bacterium powder is diluted to containing bacterium number by 2 fermented and cultureds6The bacterium solution of CFU/mL uses liquid amplification cultivation
Base, amplification cultivation 48 hours (3 parallel) at 37 DEG C.Detect the Lactobacillus crispatus viable count in culture solution.
3 results are through 48h amplification cultivation, and viable count is significantly higher than Lactobacillus crispatus in Lactobacillus crispatus DH200 culture solution
Viable count (P < 0.05) in DH144, Lactobacillus crispatus DH134 and commercially available Lactobacillus crispatus culture solution, is shown in Table 1.
1 Lactobacillus crispatus viable count (Log of table10CFU·mL-1, n=3)
4 discuss above-mentioned the experimental results showed that Lactobacillus crispatus DH200 is obviously compared with other bacterial strains and commercially available under equal conditions
Strain growth is vigorous.Probiotic active is high, breeding is its basis for playing prebiotic effect and guarantee fastly.The experimental results showed that curling
Lactobacillus DH200 has higher bioactivity compared with other Lactobacillus crispatus bacterial strains, has very high prebiotic value.
Application effect embodiment 2: inhibition of the Lactobacillus crispatus to Escherichia coli, staphylococcus aureus or Candida albicans
Effect
1 materials and methods
Melt MH culture medium, when the culture medium wait melt is cooled to 50 DEG C or so, sensitive indicator bacterium solution is added in every 200mL
0.5mL is poured into after mixing in 9cm culture dish, is placed in plate after solidification with tweezers clamping Oxford cup, is added in each Oxford cup
Enter 250 μ L Lactobacillus crispatus culture solutions (1 × 107CFU/mL), insulating box culture for 24 hours, measures antibacterial circle diameter (mm).
2 results
The antibacterial circle diameter of Lactobacillus crispatus DH200 be noticeably greater than Lactobacillus crispatus DH144, Lactobacillus crispatus DH134 and
Viable count (P < 0.05), is shown in Table 2 in commercially available Lactobacillus crispatus culture solution.
2 Lactobacillus crispatus antibacterial situation of table
3 discuss
This is the experimental results showed that Lactobacillus crispatus DH200 inhibits Escherichia coli, staphylococcus aureus or Candida albicans
Effect be better than other Lactobacillus crispatus, show that effect of the Lactobacillus crispatus DH200 in terms of the treatment diseases such as vaginitis is also shown
It writes and is better than other Lactobacillus crispatus.
Microorganism fungus kind Lactobacillus crispatus DH200 of the present invention used in implementation process is in 08 month 2018 01
Day is in China Committee for Culture Collection of Microorganisms's common micro-organisms center (in Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Institute of microbiology, the academy of sciences, state postcode 100101) preservation.Classification naming: Lactobacillus crispatus Lactobacillus
Crispatus, deposit number CGMCC No.16193.For above-mentioned strain through the microorganism Spot detection, testing result is survival.
Claims (10)
1. a kind of probiotics, it is characterised in that said preparation main active includes Lactobacillus crispatus, the newborn bar of the curling
Bacterium bag includes Lactobacillus crispatus DH200, deposit number CGMCC No.16193.
2. probiotics according to claim 1, it is characterised in that Lactobacillus crispatus refers to bion living.
3. probiotics according to claim 1, it is characterised in that the preparation includes composite medicine, health care product, food
Product or sterile products, sterile products further comprise inhibiting-bacteria preparation.
4. probiotics according to claim 1, it is characterised in that Lactobacillus crispatus has high bioactivity.
5. probiotics described in claim 1 are in preparation prevention or treatment Escherichia coli, staphylococcus aureus or white
Application in monilial infection preparation.
6. being applied by described in claim 5, it is characterised in that the probiotics inhibit Escherichia coli, golden yellow Portugal in preparation
Application in grape coccus or Candida albicans bacteria preparation.
7. application of the probiotics described in claim 1 in preparation prevention or treatment vaginitis preparation.
8. being applied by described in claim 7, it is characterised in that the vaginitis includes bacterial vaginosis BV, mycotic, senile
Vaginitis, colpomycosis further comprise monilial vaginitis.
9. probiotics according to claim 1, it is characterised in that the Lactobacillus crispatus includes Lactobacillus crispatus DH200
And its derivative strain.
10. probiotics according to claim 1, it is characterised in that the total viable count of the Lactobacillus crispatus that solid pharmaceutical preparation includes
Not less than 1 × 106CFU/g, generally 1 × 107CFU/g or more can reach 1 × 1012CFU/g or 1 × 1012CFU/g with
On；Or the total viable count of Lactobacillus crispatus that liquid preparation includes is not less than 1 × 106CFU/mL, generally 1 × 107CFU/mL with
On, it can reach 1 × 1012CFU/mL or 1 × 1012CFU/mL or more.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111518714A (en) * | 2019-12-19 | 2020-08-11 | 江南大学 | Lactobacillus crispatus capable of preventing and/or treating candida vaginitis |
| WO2021027742A1 (en) * | 2019-08-09 | 2021-02-18 | 四川厌氧生物科技有限责任公司 | Lactobacillus crispatus and application thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021027742A1 (en) * | 2019-08-09 | 2021-02-18 | 四川厌氧生物科技有限责任公司 | Lactobacillus crispatus and application thereof |
| CN111518714A (en) * | 2019-12-19 | 2020-08-11 | 江南大学 | Lactobacillus crispatus capable of preventing and/or treating candida vaginitis |
| CN111518714B (en) * | 2019-12-19 | 2022-02-01 | 江南大学 | Lactobacillus crispatus capable of preventing and/or treating candida vaginitis |
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