CN103717227A - Clostridium difficile growth inhibitor - Google Patents
Clostridium difficile growth inhibitor Download PDFInfo
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- CN103717227A CN103717227A CN201280024085.6A CN201280024085A CN103717227A CN 103717227 A CN103717227 A CN 103717227A CN 201280024085 A CN201280024085 A CN 201280024085A CN 103717227 A CN103717227 A CN 103717227A
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- clostridium difficile
- propionibacterium freudenreichii
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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
- 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
- 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
- A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
- A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
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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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
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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
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
Abstract
Provided are: a Clostridium difficile growth inhibitor which comprises, as the active ingredient, a propionic acid bacterium and/or a fermentation product thereof; and a prebiotics composition which has an effect of inhibiting the growth of Clostridium difficile. Also provided are: a Clostridium difficile growth inhibitor which comprises, as the active ingredient, 1,4-dihydroxy-2-naphthoic acid (DHNA) and/or an organic acid; a prebiotics composition which has an effect of inhibiting the growth of Clostridium difficile; and a food composition comprising the same. Also provided is a method for inhibiting the growth of Clostridium difficile, said method comprising taking or administering the aforesaid substances.
Description
Technical field
The present invention relates to clostridium difficile (Clostridium difficile, also referred to as " clostridium difficile ") antiblastic and there is prebiotics (prebiotecs) compositions of clostridium difficile inhibited proliferation and contain drink food compositions and the clostridium difficile propagation inhibition method that this prebiotic compositions forms.
Background technology
Clostridium difficile (Clostridium difficile) is that antimicrobial drug is the Main Pathogenic Bacteria of dysentery or the enteritis of inducement, and known be the pathogenic bacterium of pseudomembranous colitis.The symptom characteristic that clostridium difficile infects disease is the wide region from slight dysentery symptom to the serious symptom morbid state as lethal such as intestinal obstruction.In addition, this infection disease usually recurs, is difficult to treatment, and also very important as the pathogenic bacterium of nosocomial infection, often has the report that collective occurs in the person of the moving in intermediate court of inpatient or nursing homes.Clinical scene at this moment, treats owing to being difficult to medicament, therefore strongly needs utilize diet product or the prebiotics of use especially easily to come relief of symptoms, inhibition or prevention infection sick.
Up to the present, studied the reply way that various clostridium difficiles is infected to disease.For example, in patent documentation 1, disclose by treating the not digestible oligosaccharide that contains fructose oligosaccharides of effective dose and suppressed the sick method of mammiferous infection that clostridium difficile causes.In addition, in patent documentation 2, disclose by adding in drink Foods or drinks or giving Flos lupuli (Flos Humuli Lupuli) extract etc. and suppressed the propagation of clostridium difficile, and then the disease for the treatment of or preventing to bring out thus.In addition, a kind of turcin CD(turicine CD that is called is disclosed in patent documentation 3) bacteriocin, it is accompanied by the increase that nosocomial infection's sexuality that clostridium difficile etc. causes is caught an illness, novel antibacterial compounds as these diseases of reply, has clostridium difficile proliferation inhibiting effect.
Prior art document
Patent documentation
Patent documentation 1: No. 5688777 description of United States Patent (USP)
Patent documentation 2: Japanese kokai publication hei 11-221064 communique
Patent documentation 3: Japanese Unexamined Patent Application Publication 2011-505126 communique
Summary of the invention
The problem that invention wish solves
But, in the disclosed technology of above-mentioned patent documentation, the prebiotics and the prebiotics that when not being disclosed in the propagation that promotes useful bacillus bifidus, suppress the propagation etc. of clostridium difficile (Clostridium difficile) carry out derived components, and up to the present also do not know the example that it is reported.
Therefore, still wish infection disease or the technology to its prevention that continual exploitation can suppress the propagation of clostridium difficile (Clostridium difficile) and realize thus remission and then suppress to be caused by it with easier method and high effect.
For solving the means of problem
The present inventors are in view of above-mentioned problem in the past conducts in-depth research, and found that the culture supernatant of propionibacterium freudenreichii (Propionibacterium freudenreichii), particularly ET-3 strain or ATCC9614T strain etc. can suppress the propagation of clostridium difficile.And found the propagation that a large amount of Isosorbide-5-Nitrae-dihydroxy-2-benzoic acid (DHNA) that produce such as propionibacterium freudenreichii ET-3 strain or ATCC9614T strain and/or organic acid suppress clostridium difficile significantly.
Therefore, the invention provides do not affect the useful antibacterial such as bacillus bifidus, lactobacillus and suppress significantly clostridium difficile propagation prebiotic compositions and contain drink food compositions and clostridium difficile (Clostridium difficile) the propagation inhibition method that this prebiotic compositions forms.
The preferred mode of the present invention relates to a kind of clostridium difficile (Clostridium difficile) antiblastic, and it contains bacterium acidi propionici and/or its fermented product as effective ingredient.
In addition, a mode of the present invention relates to a kind of prebiotic compositions with clostridium difficile (Clostridium difficile) inhibited proliferation, and it contains bacterium acidi propionici and/or its fermented product forms.
As the described bacterium acidi propionici in above-mentioned antiblastic or prebiotic compositions, be preferably propionibacterium freudenreichii ET-3(Propionibacterium freudenreichii ET-3) or propionibacterium freudenreichii ATCC9614T(Propionibacterium freudenreichii ATCC9614T).
The preferred mode of the present invention relates to a kind of clostridium difficile (Clostridium difficile) antiblastic, and it contains Isosorbide-5-Nitrae-dihydroxy-2-benzoic acid (DHNA) and/or organic acid as effective ingredient.
In addition, another way of the present invention relates to a kind of prebiotic compositions with clostridium difficile (Clostridium difficile) inhibited proliferation, and it contains Isosorbide-5-Nitrae-dihydroxy-2-benzoic acid (DHNA) and/or organic acid forms.
As described 1 in above-mentioned antiblastic or prebiotic compositions, 4-dihydroxy-2-benzoic acid (DHNA), is preferably the ET-3 by propionibacterium freudenreichii ET-3(Propionibacterium freudenreichii) or propionibacterium freudenreichii ATCC9614T(Propionibacterium freudenreichii ATCC9614T) the DHNA that obtains of fermented product or be synthetic DHNA.DHNA can be by well known to a person skilled in the art that maneuver suitably synthesizes.And described organic acid can list acetic acid, formic acid, propanoic acid etc., is particularly preferably propanoic acid.
A mode more of the present invention relates to a kind of drink food compositions with clostridium difficile (Clostridium difficile) inhibited proliferation, and it is that the bacterium acidi propionici and/or its fermented product that contain effective dose form.
As the described bacterium acidi propionici in above-mentioned drink food compositions, be preferably propionibacterium freudenreichii ET-3(Propionibacterium freudenreichii ET-3) or propionibacterium freudenreichii ATCC9614T(Propionibacterium freudenreichii ATCC9614T).
In addition, another mode of the present invention relates to a kind of drink food compositions with clostridium difficile (Clostridium difficile) inhibited proliferation, it is that the Isosorbide-5-Nitrae-dihydroxy-2-benzoic acid (DHNA) and/or the organic acid that contain effective dose form.
As described 1 in above-mentioned drink food compositions, 4-dihydroxy-2-benzoic acid (DHNA), preferably by propionibacterium freudenreichii ET-3(Propionibacterium freudenreichii ET-3) or propionibacterium freudenreichii ATCC9614T(Propionibacterium freudenreichii ATCC9614T) the DHNA that obtains of fermented product.
Another mode of the present invention relates to a kind of clostridium difficile (Clostridium difficile) propagation inhibition method, it is characterized in that, absorbs or prebiotic compositions that the bacterium acidi propionici that contains effective dose and/or its fermented product form.
Described bacterium acidi propionici as in above-mentioned clostridium difficile (Clostridium difficile) propagation inhibition method, is preferably propionibacterium freudenreichii ET-3(Propionibacterium freudenreichii ET-3) or propionibacterium freudenreichii ATCC9614T(Propionibacterium freudenreichii ATCC9614T).
In addition, another mode of the present invention relates to a kind of clostridium difficile (Clostridium difficile) propagation inhibition method, it is characterized in that the prebiotic compositions that the Isosorbide-5-Nitrae-dihydroxy that absorbs or contain effective dose-2-benzoic acid (DHNA) and/or organic acid form.As giving object, can list such as people or non-human mammal etc.In addition, above-mentioned propagation inhibition method of the present invention comprises for example method of interior (in vivo), external (in vitro) of body, in vitro (ex vivo).
As described 1 in above-mentioned clostridium difficile (Clostridium difficile) propagation inhibition method, 4-dihydroxy-2-benzoic acid (DHNA), is preferably the ET-3 by propionibacterium freudenreichii ET-3(Propionibacterium freudenreichii) or propionibacterium freudenreichii ATCC9614T(Propionibacterium freudenreichii ATCC9614T) the DHNA that obtains of fermented product.
In addition, clostridium difficile antiblastic of the present invention as described later, has the useful antibacterial such as bacillus bifidus, lactobacillus is not had to the effect that affects and significantly suppress the propagation of clostridium difficile.Therefore, as the mode of utilizing of antiblastic of the present invention, can list such as with the propagation of the useful antibacterials such as bacillus bifidus, lactobacillus is there is to the antibiotic of influence or the mode of antimicrobial drug use.That is, antiblastic of the present invention as with antibiotic or antimicrobial drug and with agent, be useful.
As can with the antibiotic of antiblastic of the present invention use, can list such as lincomycin is that antibiotic, beta-lactam are that antibiotic, aminoglycoside are that antibiotic, macrolide antibiotic, terracycline antibiotics, chloromycetin are that antibiotic, peptide are antibiotic, fosfomycin etc.; As antimicrobial drug of the present invention, can list such as sulphonamides, antitubercular agent, quinolinone is that antimicrobial drug, new quinolinone are antimicrobial drug, antiviral agents, antifungal agent etc.These antibiotic or antimicrobial drug, owing to may causing the propagation of clostridium difficile, therefore should come and use for the object that suppresses the propagation of clostridium difficile.In addition, metronidazole (metronidazole), vancomycin (vancomycin), feldamycin (Fidaxomicin) etc. to the effective antibiotic of clostridium difficile with the form of supplementary function and clostridium difficile antiblastic of the present invention and with being also contained in scope of the present invention.
In addition, the invention still further relates to bacterium acidi propionici and/or its fermented product purposes in manufacturing clostridium difficile (Clostridium difficile) antiblastic.
In addition, the invention still further relates to Isosorbide-5-Nitrae-dihydroxy-2-benzoic acid (DHNA) and/or the organic acid purposes in manufacturing clostridium difficile (Clostridium difficile) antiblastic.
In addition, the invention still further relates to bacterium acidi propionici and/or its fermented product suppressing for clostridium difficile (Clostridium difficile) propagation.
In addition, the invention still further relates to the Isosorbide-5-Nitrae-dihydroxy-2-benzoic acid (DHNA) suppressing for clostridium difficile (Clostridium difficile) propagation.
Invention effect
Prebiotic compositions of the present invention promotes the propagation as the bacillus bifidus of the useful antibacterial of enteral, and does not affect the propagation of lactobacillus.Therefore, the prebiotic compositions of the present invention that contains propionibacterium freudenreichii ET-3 strain and ATCC9614T strain and DHNA etc. not only can promote the propagation of bacillus bifidus etc., and can suppress the propagation as the clostridium difficile (Clostridium difficile) of extremely harmful antibacterial.Therefore, the present invention can provide and have the prebiotic compositions of very useful clostridium difficile inhibited proliferation and contain drink food compositions and the clostridium difficile propagation inhibition method that this prebiotic compositions forms.
Accompanying drawing explanation
Fig. 1 is for representing the curve chart of prebiotic compositions of the present invention to the proliferation inhibition activity of clostridium difficile.
Fig. 2 is for representing the curve chart of prebiotic compositions of the present invention to the proliferation inhibition activity of clostridium difficile.
Fig. 3 is for representing the curve chart of prebiotic compositions of the present invention on the impact of the propagation of bacillus bifidus (Bifidobacterium longum OLB6001 strain).
Fig. 4 is for representing the curve chart of prebiotic compositions of the present invention on the impact of the propagation of lactobacillus (L.bulgaricus).
Fig. 5 is for representing the curve chart of prebiotic compositions of the present invention on the impact of the propagation of lactobacillus (S.thermophilus OLS3059 strain).
Fig. 6 is for representing the curve chart of propanoic acid of the present invention on the impact of the propagation of clostridium difficile.
Fig. 7 is deposit number FERM BP-8115(propionibacterium freudenreichii ET-3 strain) the copy of preservation proof.
Fig. 8 is deposit number FERM BP-8115(propionibacterium freudenreichii ET-3 strain) the copy of survival proof.
The specific embodiment
Below the present invention is at length described, but the present invention is not subject to the restriction of each mode of the following stated.
Bacterium acidi propionici is used as the leaven that Ai Manta draws cheese etc. from ancient times, and its safety is firmly established.Example as the bacterium acidi propionici containing in clostridium difficile antiblastic of the present invention or prebiotic compositions, can list propionibacterium freudenreichii ET-3(Propionibacterium freudenreichii ET-3), propionibacterium freudenreichii ATCC9614T(Propionibacterium freudenreichii ATCC9614T), propionibacterium freudenreichii IFO12424(Propionibacterium freudenreichii IFO12424), propionibacterium jensenii (Propionibacterium jensenii), product propionibacterium acide-propionici (Propionibacterium acidipropionici) etc. belongs to the microorganism of propionibacterium.Wherein, particularly preferably propionibacterium freudenreichii ET-3 strain, propionibacterium freudenreichii ATCC9614T strain.
These thalline can also can be used by multiple combination separately.
As the bacterium acidi propionici using in clostridium difficile antiblastic of the present invention or prebiotic compositions and/or its fermented product, can be by described propionibacterium microorganism belonging to genus be cultivated and is obtained according to conventional method.The fermented product obtaining itself and/or its handled thing can be used as to antiblastic of the present invention or prebiotic compositions.Handled thing can use such as the fermented product that contains thalline, the filtrate that fermented product filtration or degerming are obtained, with specific solvent extract and the extract that obtains, concentrate fermented product, its filtrate, extract, supernatant concentration being obtained with vaporizer etc., they the process treatment process such as dry thing (lyophilization or other) and the handled thing that obtains.
As the addition of preferred bacterium acidi propionici and/or its fermented product, according to the difference of enrichment of fermented product etc. and difference.For example, in the situation that the fermented product of the bacterium acidi propionici of the method manufacture of recording in the embodiment 1 by International Publication pamphlet WO2005/033323 description is concentrated into DHNA concentration, reach 100 μ g/ml, its addition is 0.1～10 % by weight, is preferably 0.3～5 % by weight with respect to antiblastic of the present invention or prebiotic compositions.
As in clostridium difficile antiblastic of the present invention or prebiotic compositions, contain 1,4-dihydroxy-2-benzoic acid (DHNA), can use any in the DHNA obtaining by chemosynthesis or biosynthesis, be preferably the DHNA obtaining by biosynthesis.In the present invention, particularly preferably use propionibacterium freudenreichii (Propionibacterium freudenreichii) ET-3 strain or propionibacterium freudenreichii ATCC9614T(Propionibacterium freudenreichii ATCC9614T) DHNA that produces in a large number.For example, can will be used as DHNA by propionibacterium microorganism belonging to genus is cultivated to the fermented product itself and/or its handled thing that obtain according to conventional method.Handled thing can use such as the fermented product that contains thalline, by fermented product filtration or degerming and the filtrate obtaining, utilize regulation solvent extraction and the rough things such as extract or their the refining thing that obtain, utilize vaporizer etc. by fermented product, its filtrate, extract, supernatant concentration and the concentrate obtaining, they the process treatment process such as dry thing (lyophilization or other) and the handled thing that obtains.As the concentration of DHNA, can suitably adjust according to purposes etc., be 1～250 μ g/ml, be preferably 1～5 μ g/ml.
The present invention also provides the drink that contains prebiotic compositions food compositions.
The drink food compositions that contains prebiotic compositions of the present invention particularly, can drink food compositions such as milk, refreshment drink, fermentation milk (Yoghourt, boruga), cheese, bread, cookies, cracknel, pizza, formula milk, liquid food, patient with food, child with the food such as milk powder, nursing women with interpolation prebiotic compositions of the present invention in the food such as milk powder, nutraceutical etc., then by its picked-up.In addition, about its character, can adopt the state of normally used diet product, such as solid, shaped (powder, graininess or other), pasty state, aqueous to the common negotiable so-called drink shape of food such as suspension, liquid food shape, fruit jelly shape, tablet shape, graininess, capsule shape.Wherein, from the angle that can absorb easily, absorbefacient angle grade, drink food compositions of the present invention preferably provides with the form of beverage.
About other compositions, be not particularly limited, in the situation that containing prebiotic compositions of the present invention at drink in food compositions and using, water, albumen, saccharic, lipid, vitamins, minerals, organic acid, organic base, fruit juice, flavoring agent class etc. can be used as main constituent.As albumen, can list of animal or plant nature albumen, their hydrolysates such as whole milk powder, defatted milk powder, partially skimmed milk powder, casein, whey powder, lactalbumin, Lactalbumin concentrate, lactalbumin isolate, alpha-casein, beta-casein, κ-casein, beta lactoglobulin, alpha-lactalbumin, lactoferrin, soybean protein, Chicken Albumin, meat albumen; The various milk-derived one-tenth such as butter, whey mineral, butter, milk surum, amino acids, sialic acid, phospholipid, lactose grade.As saccharic, can list common saccharide, producing starch (except dextrin, also having soluble starch, british starch, Oxytarch, starch ester, starch ether etc.), dietary fiber etc.As lipid, can list such as Adeps Sus domestica, fish oil etc., their separating oil, hydrogenated oil and fat, ester exchange wet goods animal raw fat; Petiolus Trachycarpi oil, safflower oil, Semen Maydis oil, Oleum Brassicae campestris, Oleum Cocois, their separating oil, hydrogenated oil and fat, ester exchange wet goods vegetative grease etc.As vitamins, can list such as vitamin A, carotenoid, vitamin B complex, vitamin C, vitamin D family, vitamin E, vitamin K family, Citrin, CoenzymeQ10, nicotinic acid, nicotinic acid, pantothenic acid, biotin, inositol, choline, folic acid etc.; As minerals, can list such as calcium, phosphorus, potassium, magnesium, chlorine, sodium, ketone, ferrum, manganese, zinc, selenium, chromium, molybdenum, vanadium, whey mineral etc.As organic acid, can list such as malic acid, citric acid, lactic acid, tartaric acid etc.These compositions can also can two or more be used in combination separately, can use composite and/or contain in a large number their diet product.
Above-mentioned drink food compositions can be by mixing prebiotic compositions of the present invention according to conventional method and manufacture with the appropriate amount of above-mentioned neccessary composition.
The content of the prebiotic compositions in drink food compositions of the present invention can at random be determined according to its object, purposes etc.The present invention is not limited thereto, as its content, with respect to total amount, more than being generally 5 % by weight, more preferably more than 10 % by weight.By by its picked-up or give, can carry out easily clostridium difficile (Clostridium difficile) propagation and suppress, even can relief of symptoms, inhibition or prevention infection sick.
It should be noted that, whole prior art documents of quoting in this description are included in this description by reference.
Embodiment
Below enumerate embodiment the present invention is described in further detail, but the present invention is not limited thereto.Wherein, in embodiment, the statement of " % " represents % by weight except the situation of special instruction.
[embodiment 1]
The preparation of the bacterium acidi propionici culture supernatant as prebiotic compositions of the present invention
By propionibacterium freudenreichii ET-3 strain (be preserved in Japanese Independent Administrative Leged Industrial Technology Complex Inst patent biological preservation center (day national thatch city Ken つく ば city East 1 fourth order a kind ground 1 central 6(Postal just designation 305-8566) with FERM BP-8115 August 9 calendar year 2001), propionibacterium freudenreichii ATCC9614T strain (propionibacterium freudenreichii benchmark strain, from American Type Culture Collection(ATCC) buy), from commercially available Ai Manta, draw cheese the separated propionibacterium freudenreichii strain (No.32-1) obtaining to activate under 30 ℃ of anaerobic conditions and preculture 48 hours, then further under 30 ℃ of anaerobic conditions, cultivate 72 hours.By culture centrifugalize (centrifugalize condition: 8000rpm, 4 ℃, 10 minutes), reclaim culture supernatant, filter sterilised, using the culture supernatant obtaining as prebiotic compositions of the present invention, is kept at-80 ℃.
The cultivation of clostridium difficile
Clostridium difficile JCM1296T strain (strain of clostridium difficile benchmark) is buied for grinding BioResource Center from reason, according to appended handbook, restores.
Inhibition test
The clostridium difficile of as above preparation is inoculated in the BHI culture medium that keeps in advance anaerobic state with 1%.Add wherein prebiotic compositions of the present invention or DHNA, under 37 ℃ of anaerobic conditions, cultivate 24 hours.After culture fluid is fully stirred, measure OD
650(Shimadzu spectrophotometer BioSpec1600, Shimadzu Scisakusho Ltd's system).Result is made to curve chart and be shown in Fig. 1.Wherein, to the data that obtain, utilize Tukey-Kramer method to carry out multiple comparisons check, each organizes n=3, * is that p<0.05, * * are p<0.01.Wherein, propionibacterium freudenreichii ATCC9614T and propionibacterium freudenreichii ET-3 are p<0.01 with respect to matched group.
[result]
In having added the system of 5% prebiotic compositions of the present invention, the OD after 24 hours cultivating
650significantly reduce the propagation of hence one can see that prebiotic compositions of the present invention suppresses effectively clostridium difficile.
[embodiment 2]
The preparation of contained DHNA in prebiotic compositions of the present invention
By DHNA(Sigma-Aldrich system) 100mg is dissolved in the DMSO of 0.5ml, makes the DMSO solution of 1M.With 10 times of BHI culture medium dilutions, make 100mM, then filter sterilised.Meanwhile, by 10 times of BHI culture medium dilutions for DMSO, make solution, filter sterilised.Thereafter dilution is all carried out 10 times of dilutions with DMSO solution, makes to contain 10% DMSO in the DHNA solution of total concentration.Due to these DHNA solution of 0.1% are added to, in bacteria culture fluid, make prebiotic compositions of the present invention, so the ultimate density of DMSO is 0.01%.
Inhibition test
Proliferation inhibiting effect about prebiotic compositions of the present invention to the clostridium difficile using in embodiment 1, carries out inhibition test similarly to Example 1.Particularly, for each sample, the prebiotic compositions of the present invention that contains DHNA is added to (1 μ M, 10 μ M, 20 μ M, 50 μ M) with various concentration.The result obtaining is as shown in the curve chart of Fig. 2.Wherein, utilize Tukey-Kramer method to carry out multiple comparisons check, each organizes n=3, * is p<0.05.Wherein, for 10 μ M～50 μ M, with respect to DMSO group, for * * is p<0.01.
As shown in Figure 2, by adding DHNA more than 10 μ M, the propagation of clostridium difficile is significantly suppressed.
[embodiment 3]
The preparation of bacillus bifidus and lactobacillus culture supernatant
By from bacillus bifidus (B.longum) OLB6001 strain (FERM P-13610) and " Mingzhi bulgaria Yoghourt " (trade name: the Bulgarian Lactobacillus (L.bulgaricus) that company of VAAM system) separation obtains is cultivated 18 hours under 37 ℃ of anaerobic conditions.Streptococcus thermophilus (S.thermophilus) OLS3059 strain (FERM P-15487) is cultivated 30 hours under 30 ℃ of anaerobic conditions.By culture centrifugalize (centrifugalize condition: 8000rpm, 4 ℃, 10 minutes), reclaim culture supernatant, filter sterilised, by the culture supernatant obtaining as a comparison compositions be stored in-80 ℃ (lactobacillus is used in embodiment 4).
The impact of prebiotic compositions of the present invention on the propagation of bacillus bifidus
In bacillus bifidus (Bifidobacterium longum) OLB6001 strain, add DHNA or the 0.01%DMSO(contrast of using in embodiment 2), under 37 ℃ of anaerobic conditions, cultivate 12 hours.After culture fluid is fully stirred, measure OD
650(Shimadzu spectrophotometer BioSpec1600, Shimadzu Scisakusho Ltd's system).Result is made to curve chart and be shown in Fig. 3.Wherein, utilize Tukey-Kramer method to carry out multiple comparisons check, each organizes n=3, * is p<0.05.Wherein, for 10 μ M～50 μ M, with respect to DMSO group, for * * is p<0.01.
[result]
Known by adding the prebiotic compositions of the DHNA of containing of the present invention, the propagation of OLB6001 strain is significantly promoted.Represent, not only prebiotic compositions of the present invention does not affect useful antibacterial and has a propagation facilitation effect.Therefore known, when the propagation of carrying out clostridium difficile suppresses, on not impact (thering is on the contrary propagation facilitation effect) of the propagation of useful antibacterial.
[embodiment 4]
The impact of prebiotic compositions of the present invention on the propagation of lactobacillus
In order to study prebiotic compositions (the DHNA concentration: 1 μ M～50 μ M) on the L.bulgaricus using in embodiment 3 and the impact of S.thermophilus OLS3059 strain of the DHNA of containing of the present invention, test similarly to Example 3, show the result in Fig. 4 and Fig. 5.Known similarly to Example 3, by adding the prebiotic compositions of the DHNA of containing of the present invention, on the not impact of the propagation of L.bulgaricus and S.thermophilus OLS3059 strain.Therefore known, when the propagation of carrying out clostridium difficile suppresses, on not impact of the propagation of useful antibacterial.
[embodiment 5]
The impact of propanoic acid of the present invention on the propagation of clostridium difficile
By 1% cultivation 24 hours, OD
650reach 1.2 through propagation clostridium difficile (C.difficile JCM1296 strain) culture fluid add in culture medium, add various organic acid simultaneously, make final concentration reach 100 μ M, measure the OD after 24 hours
650value.Result is made to curve chart to be shown in shown in Fig. 6.
As shown in Figure 6, by adding propanoic acid, the propagation of clostridium difficile is inhibited.
Utilizability in industry
The propagation that clostridium difficile antiblastic of the present invention or prebiotic compositions are bacillus bifidus or lactobacillus on the useful antibacterial of enteral does not affect or is effectively to propagation, and the antibacterial that can suppress to be extremely harmful to is on the other hand the propagation of clostridium difficile (Clostridium difficile).Therefore, prebiotic compositions of the present invention based on containing propionibacterium freudenreichii ET-3 strain and ATCC9614T strain and DHNA etc., can provide and have the prebiotic compositions of very useful clostridium difficile inhibited proliferation and contain drink food compositions and the clostridium difficile propagation inhibition method that this prebiotic compositions forms, the utilizability in industry is very high.
Claims (9)
1. clostridium difficile (Clostridium difficile) antiblastic, it contains bacterium acidi propionici and/or its fermented product as effective ingredient.
2. medicament according to claim 1, wherein, described bacterium acidi propionici is propionibacterium freudenreichii ET-3(Propionibacterium freudenreichii ET-3) or propionibacterium freudenreichii ATCC9614T(Propionibacterium freudenreichii ATCC9614T).
3. clostridium difficile (Clostridium difficile) antiblastic, it contains Isosorbide-5-Nitrae-dihydroxy-2-benzoic acid is that DHNA and/or organic acid are as effective ingredient.
4. medicament according to claim 3, wherein, described Isosorbide-5-Nitrae-dihydroxy-2-benzoic acid is that DHNA is from propionibacterium freudenreichii ET-3(Propionibacterium freudenreichii ET-3) or propionibacterium freudenreichii ATCC9614T(Propionibacterium freudenreichii ATCC9614T) fermented product obtain.
With antibiotic or antimicrobial drug use and with an agent, it contains the medicament described in any one in claim 1～4.
6. clostridium difficile (Clostridium difficile) propagation inhibition method, is characterized in that, absorbs or prebiotic compositions that the bacterium acidi propionici that contains effective dose and/or its fermented product form.
7. clostridium difficile according to claim 6 (Clostridium difficile) is bred inhibition method, it is characterized in that, described bacterium acidi propionici is propionibacterium freudenreichii ET-3(Propionibacterium freudenreichii ET-3) or propionibacterium freudenreichii ATCC9614T(Propionibacterium freudenreichii ATCC9614T).
8. clostridium difficile (Clostridium difficile) propagation inhibition method, is characterized in that, the Isosorbide-5-Nitrae-dihydroxy that absorbs or contain effective dose-2-benzoic acid is the prebiotic compositions that DHNA and/or organic acid form.
9. clostridium difficile according to claim 8 (Clostridium difficile) is bred inhibition method, it is characterized in that, described Isosorbide-5-Nitrae-dihydroxy-2-benzoic acid is that DHNA is by propionibacterium freudenreichii ET-3(Propionibacterium freudenreichii ET-3) or propionibacterium freudenreichii ATCC9614T(Propionibacterium freudenreichii ATCC9614T) fermented product obtain.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2011072462 | 2011-03-29 | ||
| JP2011-072462 | 2011-03-29 | ||
| PCT/JP2012/056628 WO2012132913A1 (en) | 2011-03-29 | 2012-03-15 | Clostridium difficile growth inhibitor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103717227A true CN103717227A (en) | 2014-04-09 |
| CN103717227B CN103717227B (en) | 2016-08-17 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201280024085.6A Active CN103717227B (en) | 2011-03-29 | 2012-03-15 | Clostridium difficile growth inhibitor |
Country Status (5)
| Country | Link |
|---|---|
| JP (2) | JP6029242B2 (en) |
| CN (1) | CN103717227B (en) |
| HK (1) | HK1195874A1 (en) |
| TW (1) | TW201245441A (en) |
| WO (1) | WO2012132913A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109803668A (en) * | 2016-10-14 | 2019-05-24 | 株式会社明治 | For inhibiting the composition of lactobacillus lactic acid bacteria reduction in enteron aisle |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040170617A1 (en) * | 2000-06-05 | 2004-09-02 | Finegold Sydney M. | Method of treating diseases associated with abnormal gastrointestinal flora |
| WO2006098447A1 (en) * | 2005-03-18 | 2006-09-21 | Meiji Feed. Co., Ltd. | Feeding stuff composition comprising proliferation promoting agent for bifidus bacteria and use of the same |
| JP2010280582A (en) * | 2009-06-02 | 2010-12-16 | Miyarisan Pharmaceutical Co Ltd | Anti-helicobacter agent |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101155735B1 (en) * | 2004-03-31 | 2012-06-12 | 가부시키가이샤 메이지 | Antibacterial compositions |
-
2012
- 2012-03-15 JP JP2013507369A patent/JP6029242B2/en active Active
- 2012-03-15 CN CN201280024085.6A patent/CN103717227B/en active Active
- 2012-03-15 WO PCT/JP2012/056628 patent/WO2012132913A1/en active Application Filing
- 2012-03-28 TW TW101110734A patent/TW201245441A/en unknown
-
2014
- 2014-09-16 HK HK14109329.7A patent/HK1195874A1/en not_active IP Right Cessation
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2016
- 2016-08-30 JP JP2016167531A patent/JP2017019833A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040170617A1 (en) * | 2000-06-05 | 2004-09-02 | Finegold Sydney M. | Method of treating diseases associated with abnormal gastrointestinal flora |
| WO2006098447A1 (en) * | 2005-03-18 | 2006-09-21 | Meiji Feed. Co., Ltd. | Feeding stuff composition comprising proliferation promoting agent for bifidus bacteria and use of the same |
| JP2010280582A (en) * | 2009-06-02 | 2010-12-16 | Miyarisan Pharmaceutical Co Ltd | Anti-helicobacter agent |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109803668A (en) * | 2016-10-14 | 2019-05-24 | 株式会社明治 | For inhibiting the composition of lactobacillus lactic acid bacteria reduction in enteron aisle |
| TWI745454B (en) * | 2016-10-14 | 2021-11-11 | 日商明治股份有限公司 | Composition for inhibiting the reduction of Lactobacillus spp. lactic acid bacteria in the intestinal tract |
| CN109803668B (en) * | 2016-10-14 | 2024-01-09 | 株式会社明治 | Composition for inhibiting the reduction of Lactobacillus in the intestinal tract |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2017019833A (en) | 2017-01-26 |
| JP6029242B2 (en) | 2016-11-24 |
| TW201245441A (en) | 2012-11-16 |
| JPWO2012132913A1 (en) | 2014-07-28 |
| TWI560270B (en) | 2016-12-01 |
| CN103717227B (en) | 2016-08-17 |
| HK1195874A1 (en) | 2014-11-28 |
| WO2012132913A1 (en) | 2012-10-04 |
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