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WO2018168821A1 - Rince-bouche - Google Patents

Rince-bouche Download PDF

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Publication number
WO2018168821A1
WO2018168821A1 PCT/JP2018/009638 JP2018009638W WO2018168821A1 WO 2018168821 A1 WO2018168821 A1 WO 2018168821A1 JP 2018009638 W JP2018009638 W JP 2018009638W WO 2018168821 A1 WO2018168821 A1 WO 2018168821A1
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WIPO (PCT)
Prior art keywords
periodontal disease
bacteria
bean extract
periodontal
cocoa bean
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Ceased
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PCT/JP2018/009638
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English (en)
Japanese (ja)
Inventor
信城 浜田
悠 佐々木
清子 渡辺
文彦 吉野
彩佳 吉田
正樹 寺原
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Meiji Co Ltd
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Meiji Co Ltd
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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 OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • 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
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • 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)
    • 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)
    • A61K36/577—Malvaceae (Mallow family)
    • A61K36/5777—Theobroma, e.g. cocao or cocoa
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • 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
    • A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q11/00—Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses

Definitions

  • the present invention relates to an oral composition, and more particularly to an oral composition that is effective for inhibiting the adhesion of periodontal bacteria to animal cells and suppressing the expression of pili of periodontal bacteria.
  • Periodontal disease is a disease of periodontal tissue caused by periodontal bacteria such as Porphyromonas gingivalis. Plaques formed by periodontal bacteria cause inflammation in the periodontal tissue, eventually melting the bones that support the teeth and eventually causing tooth loss. It is known that in the case of mild periodontal disease, inflammation is improved by sterilizing and removing periodontal bacteria and removing plaque, and periodontal tissue returns to a healthy state. Therefore, it can be said that prevention and initial treatment are important for periodontal disease, and various bactericides and antibacterial agents effective for prevention and initial treatment of periodontal disease have been developed.
  • Non-Patent Document 1 cocoa extract and cocoa mass extract
  • Patent Documents 1 to 3 A cacao husk extract (Non-Patent Documents 1 to 3) derived from a product is known. It is known that various components contained in cocoa beans are changed by fermentation (Non-patent Document 4), but cocoa mass and cocoa extract made from fermented cocoa beans have been reported to have a bactericidal effect on periodontal disease bacteria. On the other hand, the bactericidal effect of periodontal disease bacteria has not been reported so far for extracts from unfermented cocoa beans.
  • Non-patent Document 5 No inhibitory effect on pili expression or adhesion to animal epithelial cells has been reported so far.
  • An object of the present invention is to provide a novel oral composition, in particular, an oral composition that is effective in inhibiting the adhesion of periodontal disease bacteria to animal cells and suppressing the development of periodontal disease bacteria.
  • Another object of the present invention is to provide an oral composition effective against periodontal disease bacteria and bad breath-causing bacteria.
  • the present inventors have now demonstrated that unfermented cocoa bean extract suppresses the pili expression of periodontal disease bacteria, and that unfermented cocoa bean extract and fermented cocoa bean extract adhere to animal cells of periodontal disease bacteria. Found to inhibit.
  • the present inventors also found that the unfermented cacao bean extract has excellent antibacterial activity against periodontal disease bacteria, and has excellent antibacterial activity against biofilms formed by periodontal disease bacteria. .
  • the present inventors have further found that periodontitis is improved when an unfermented cocoa bean extract is administered to a periodontitis model rat.
  • the present invention is based on these findings.
  • An oral composition comprising a cocoa bean extract.
  • the composition described in [1] above which is an inhibitor of periodontal disease bacteria adhesion to animal cells.
  • the composition according to the above [1] which is a pili expression inhibitor of periodontal disease bacteria.
  • the composition according to [1] above, wherein the cocoa bean extract is an unfermented cocoa bean extract.
  • the composition according to [4] above which is an antibacterial agent against periodontal disease bacteria and / or halitosis-causing bacteria.
  • the composition according to [4] or [5] above which is used for treatment, prevention or improvement of periodontal disease and related diseases.
  • composition according to the above [4] or [5] for use in suppressing bad breath [8]
  • Periodontopathic fungi are Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Treponema denticola, Tannerera forcicensis, Aggregatebacter actinomycetemcomitans, Campylobacter rectus and Florobacteria
  • the bacteria according to [5] above, wherein the bad breath causing bacteria is one or more selected from the group consisting of Fusobacterium nucleatum and Beironella disper and periodontal disease bacteria described in [8] above.
  • Composition [10] The composition according to any one of [1] to [9] above, which is a food composition.
  • a method for inhibiting the attachment of periodontal bacteria to animal cells and the pili of periodontal pathogens comprising ingesting or administering an effective amount of cacao bean extract to a subject in need thereof Expression suppression method.
  • a method for sterilizing periodontal disease bacteria and / or halitosis-causing bacteria comprising ingesting or administering an effective amount of an unfermented cacao bean extract to a subject in need thereof.
  • a method for treating, preventing or ameliorating periodontal disease and related diseases comprising ingesting or administering an effective amount of an unfermented cocoa bean extract to a subject in need thereof.
  • Inhibition of periodontal pathogens to animal cells and suppression of periodontal pathogens for the production of periodontal pathogens and inhibitors of periodontal pathogens Use of cacao bean extract as an inhibitor for adhesion to animal cells of periodontal disease bacteria and as an inhibitor of pili expression of periodontal disease bacteria.
  • a cacao bean extract for use in inhibiting adhesion of periodontal disease bacteria to animal cells or for use in suppressing the expression of pili of periodontal disease bacteria.
  • An unfermented cacao bean extract for use in sterilization of periodontal disease bacteria and / or halitosis-causing bacteria.
  • An unfermented cocoa bean extract for use in the treatment, prevention or amelioration of periodontal disease and related diseases.
  • the oral composition of the present invention contains a cacao bean extract as an active ingredient. Since the cocoa bean extract is a natural material that humans have used as a raw material for food for many years, according to the present invention, there is no concern about side effects, and a safe composition for oral cavity can be provided.
  • the oral composition of the present invention comprises a cocoa bean extract.
  • cocoa bean extract is used to mean not only fermented cocoa bean extract but also unfermented cocoa bean extract.
  • the oral composition of the present invention comprises a fermented cocoa bean extract.
  • a fermented cocoa bean extract a raw material obtained by processing a fermented cocoa bean can be used as it is, or it can be used as a raw material for an extraction process described later.
  • the processed material include those obtained by removing the skin from a cocoa bean using a separator, such as cocoa nibs, cocoa mass, defatted cocoa mass, and cocoa powder. That is, in the present invention, cacao nibs, cacao mass, defatted cacao mass, cocoa powder, and extracts thereof can be used as fermented cacao bean extracts.
  • the fermentation treatment of cocoa beans can be carried out by natural fermentation according to a conventional method.
  • Fermented cocoa bean extract can be prepared by solvent extraction of fermented cocoa beans (including those processed above).
  • Solvents that can be used include water, alcohols having 1 to 6 carbon atoms (for example, ethanol), aldehydes and ketones, and a solvent suitable for the variety of cocoa beans may be used as an extraction solvent in combination.
  • an oil extraction (expeller) step may be performed prior to the solvent extraction step.
  • a degreased extract can be obtained by carrying out the oil extraction step.
  • the extract obtained by the solvent extraction step may be filtered and concentrated, and then dried by a drying means such as a spray drying method or a freeze drying method to form a dry powder.
  • a method for preparing a fermented cocoa bean extract is known and can be prepared, for example, according to the description in JP-A-2001-69946.
  • the oral composition of the present invention comprises an unfermented cocoa bean extract.
  • An unfermented cocoa bean extract can be prepared by solvent extraction of cocoa beans (raw beans). Solvents that can be used include water, alcohols having 1 to 6 carbon atoms (for example, ethanol), aldehydes and ketones, and a solvent suitable for the variety of cocoa beans may be used as an extraction solvent in combination.
  • Solvents that can be used include water, alcohols having 1 to 6 carbon atoms (for example, ethanol), aldehydes and ketones, and a solvent suitable for the variety of cocoa beans may be used as an extraction solvent in combination.
  • a step of crushing cocoa beans or an oil extraction (expeller) step may be performed prior to the solvent extraction step.
  • a degreased extract can be obtained by carrying out the oil extraction step.
  • the extract obtained by the solvent extraction step may be filtered and concentrated, and then dried by a drying means such as a spray drying method or a freeze drying method to form a dry powder.
  • a method for preparing an unfermented cacao bean extract is known and can be prepared, for example, according to the description in JP-A No. 2014-118369.
  • unfermented cocoa bean extracts are commercially available.
  • “cocoa bean extract CBP” and “HP cocoa extract PW” manufactured by Meiji Co., Ltd. can be used in the present invention.
  • the content of the cocoa bean extract in the composition of the present invention includes the activity of inhibiting the adhesion of periodontal pathogens to animal cells, the activity of inhibiting periodontal pathogen expression, periodontal pathogens, and bad breath
  • the upper limit can be 100% by mass, 30% by mass or 10% by mass
  • the lower limit can be 0.01% by mass, 0.1% by mass. 1% by mass or 5% by mass.
  • the composition of the present invention can be used as a composition for oral cavity. Specifically, it is an inhibitor for adhesion of periodontal bacteria to animal cells (especially epithelial cells) and pili of periodontal bacteria (especially Mfa1 line). Hair and FimA pili).
  • the composition according to the invention can also be used as an antibacterial agent against periodontal bacteria and / or halitosis-causing bacteria.
  • periodontal disease bacteria include Porphyromonas bacteria such as Porphyromonas gingivalis , Prevotella intermedia , Prevotella nigrescens and other Prevotella bacteria, Treponema Dentikora (Treponema denticola), Tan'nera forsythensis (Tannella forsythensis), Aggregatibacter actinomycetemcomitans (Aggregatibacter actinomycetemcomitans), Campylobacter rectus (Campylobacter rectus), and the like Fusobacterium nucleatum (Fusobacterium nucleatum) .
  • Porphyromonas bacteria such as Porphyromonas gingivalis , Prevotella intermedia , Prevotella nigrescens and other Prevotella bacteria, Treponema Dentikora (Treponema denticola), Tan'nera forsythensis (
  • Fusobacterium genus bacteria such as Fusobacterium nucleatum (Fusobacterium nucleatum), Beironera-disper (Veillonella dispar) Beironera bacteria such as, more like the above-mentioned various periodontal bacteria.
  • composition of the present invention it is possible to inhibit periodontal disease bacteria from attaching to animal cells (especially epithelial cells), and periodontal disease fungi (especially Mfa1 and FimA pili). Expression can be suppressed.
  • the colonization of periodontal disease bacteria in the oral cavity is the basis for adhesion to cells, and cilia that are expressed on the cells are important for colonization of periodontal disease bacteria in the oral cavity and the formation of biofilms. Playing a role. Therefore, according to the composition of the present invention, it is possible to treat, prevent, and improve periodontal disease and related diseases through inhibition of periodontal disease bacteria adhesion to animal cells and suppression of periodontal disease bacteria pili expression. .
  • the total number of oral pathogens such as periodontal disease bacteria and bad breath-causing bacteria can be reduced, or the growth can be suppressed, and thus oral cavity such as periodontal disease and bad breath can be suppressed.
  • the disease can be treated, prevented or ameliorated. That is, the composition of this invention can be used for bad breath suppression, and can be used as a bad breath suppressant.
  • bad breath refers to what a person feels a bad breath component contained in exhaled breath, and is a general term for malodors emitted from the oral cavity together with exhaled breath.
  • Representative components of the malodor include volatile sulfur-containing compounds such as hydrogen sulfide and methyl mercaptan.
  • composition of the present invention can be used for the treatment, prevention or improvement of periodontal disease and related diseases. That is, according to the present invention, a therapeutic, preventive or ameliorating agent for periodontal disease and related diseases comprising unfermented cacao bean extract as an active ingredient is provided.
  • gingivitis gingivitis and periodontitis.
  • periodontal disease-related disease means a secondary disease that can occur due to periodontal disease.
  • vascular disorder for example, aspiration pneumonia, bacterial invasion into the living body
  • Angina pectoris myocardial infarction
  • premature birth diabetes and the like.
  • the form and shape are not particularly limited, and even in a solid form, liquid, semi-liquid, gel, It may be in the form of a paste or powder.
  • the composition of the present invention include food compositions such as tablets (for example, troches and chewable tablets), gums, gummi, candy, tablet confectionery, beverages, jelly, edible films, dentifrices, mouthwashes, Examples include oral care compositions such as gargles, denture cleaning agents, orally disintegrating tablets, and gels, and tablets, gums, gummi, and candy are preferred from the viewpoints of portability and retention in the oral cavity.
  • the composition of this invention can mix
  • composition of the present invention is provided as a tablet, in addition to the cocoa bean extract, sugar, excipient, sweetener, acidulant, flavor, colorant, preservative, stabilizer, emulsifier, plant extraction
  • sugar, excipient, sweetener, acidulant, flavor, colorant, preservative, stabilizer, emulsifier, plant extraction One or two or more components selected from foods, fruit juice powders and the like can be blended and manufactured by conventional methods.
  • Cocoa bean extract itself is a material that can be eaten and has antibacterial activity against periodontal disease bacteria and halitosis-causing bacteria as shown in the examples below, so that it can be provided as a pharmaceutical and taken as a daily food or supplement. It can also be provided in food. That is, the composition of the present invention can be provided as a pharmaceutical composition or a food composition. When the composition of the present invention is provided as a food composition, the composition of the present invention can be produced according to a normal production procedure of the food except that the cacao bean extract which is an active ingredient is blended.
  • the cocoa bean extract that is the active ingredient of the present invention can be prepared by containing the cocoa bean extract as it is in the food, and the food composition contains the cocoa bean extract. It is a food containing an effective amount.
  • “containing an effective amount” of the cocoa bean extract means such a content that the cocoa bean extract is ingested within a predetermined range when an amount normally consumed in each food is ingested.
  • the aspect which adds the cocoa bean extract itself which is an active ingredient of this invention, or the composition containing the same to foodstuffs (for example, drink) in a consumer, and makes it a food composition is also contained in the scope of the present invention. Shall.
  • the composition of the present invention comprises a cocoa bean extract with a single intake that is effective for the activity of inhibiting the adhesion of periodontal bacteria to animal cells, the activity of inhibiting the expression of cilia of periodontal bacteria and the antibacterial activity against periodontal bacteria. It can be provided as a composition comprising.
  • the amount of cocoa bean extract that is effective for these activities and the number of intakes per day can be determined depending on the age, symptoms, and body sensation of the user. For example, the amount of cocoa bean extract ingestion that is effective for the periodontal pathogen adhesion inhibitory activity, periodontal pathogen ciliary expression suppression activity and periodontal pathogen antibacterial activity is solid content.
  • the converted mass is 0.1 mg to 10 mg / dose, preferably 1.0 mg to 10 mg / dose.
  • the number of intakes of the cocoa bean extract effective per day for these activities is 1 to 10 times / day, preferably 3 to 5 times / day.
  • a method for inhibiting periodontal pathogen adhesion to animal cells comprising feeding or administering an effective amount of cocoa bean extract to a human or non-human animal, and a tooth.
  • a method for suppressing ciliated expression of peribacterium there is also provided a method for sterilizing periodontal disease bacteria and / or halitosis-causing bacteria, comprising ingesting or administering an effective amount of an unfermented cacao bean extract to a human or non-human animal.
  • the sterilization method, inhibition method and suppression method of the present invention can be carried out according to the description relating to the composition of the present invention.
  • treatment of periodontal disease and related diseases comprising ingesting or administering an effective amount of unfermented cocoa bean extract to a human or non-human animal.
  • a method for prevention or amelioration is provided.
  • the treatment, prevention or amelioration method of the present invention can be carried out according to the description relating to the composition of the present invention.
  • a cacao bean extract for the production of an inhibitor of periodontal pathogen adherence to animal cells and a ciliated expression inhibitor of periodontal pathogen, and periodontal pathogen Of cacao bean extract to inhibit the adhesion of animal periodontium to animal cells and to suppress the expression of fimbria of periodontal pathogens, as an inhibitor of periodontal pathogens to animal cells and as an inhibitor of periodontal pathogens Of cocoa bean extract.
  • the invention also provides the use of an unfermented cocoa bean extract for the production of antibacterial agents against periodontal and / or halitosis-causing bacteria and for the sterilization of periodontal and / or halitosis-causing fungi.
  • unfermented cocoa bean extract and the use of unfermented cocoa bean extract as an antibacterial agent against periodontal and / or halitosis-causing bacteria.
  • the use of an unfermented cocoa bean extract for the manufacture of an agent for the treatment, prevention or amelioration of periodontal disease and related diseases, and the treatment, prevention or prevention of periodontal disease and related diseases.
  • the use of unfermented cocoa bean extract for improvement and the use of unfermented cocoa bean extract as a treatment, prevention or amelioration agent for periodontal disease and related diseases is provided.
  • the use according to the invention can be carried out as described for the compositions according to the invention.
  • a cocoa bean extract for use in inhibiting adhesion of periodontal disease bacteria to animal cells or for suppressing the expression of periodontal disease bacteria.
  • the present invention also provides an unfermented cocoa bean extract for use in sterilization of periodontal disease bacteria and / or halitosis-causing bacteria.
  • the present invention also provides an unfermented cocoa bean extract for use in the treatment, prevention or amelioration of periodontal disease and related diseases.
  • the cocoa bean extract and unfermented cocoa bean extract of the present invention can be carried out as described for the composition of the present invention.
  • non-therapeutic means not including the act of operating, treating, or diagnosing a human (that is, a medical act on a human), and specifically, receiving a doctor or doctor's instructions It means that the person does not include a method of performing surgery, treatment or diagnosis on humans.
  • Example 1 Effect of unfermented cocoa bean extract on the expression of fimbria of periodontal disease bacteria (1) Preparation of unfermented cocoa bean extract-containing medium As a test sample, cocoa bean extract (polyphenol concentration of 82% by mass or less) “CBP” (manufactured by Meiji Co., Ltd.) and HP cacao extract PW (polyphenol concentration 17% by mass, hereinafter referred to as “HP-PW”) (manufactured by Meiji Co., Ltd.) were used. CBP and HP-PW are unfermented cocoa bean extracts with different extraction methods.
  • BHI-YHK medium is a brain heart infusion medium (Becton Dickinson, hereinafter referred to as “BHI medium”), 5 mg / mL yeast extract (Becton Dickinson), 5 ⁇ g / mL hemin (Wako Pure Chemical Industries, Ltd.) And 0.5 ⁇ g / mL vitamin K 1 (manufactured by Wako Pure Chemical Industries, Ltd.) was added (hereinafter the same).
  • CBP is contained by adding CBP-containing 50% ethanol solution and HP-PW-containing 50% ethanol solution to the BHI-YHK medium so that the CBP and HP-PW concentrations shown in Table 1 are obtained.
  • BHI-YHK medium and BHI-YHK medium containing HP-PW were prepared.
  • As a control group a test group was prepared in which neither CBP-containing 50% ethanol solution nor HP-PW-containing 50% ethanol solution was added to BHI-YHK medium.
  • Porphyromonas gingivalis ATCC 33277 (hereinafter the same) was used as a periodontal disease bacteria.
  • Anaerobox (Tacher-type Anaero Box ANX-3, Inc.) filled with anaerobic gas (70% N 2 , 15% H 2 , 15% CO 2 ) by inoculating Porphyromonas gingivalis into BHI-YHK medium And cultured at 37 ° C. for 18 hours. After culturing, the cells were collected by centrifugation, and a Porphyromonas gingivalis bacterial solution having an absorbance of 1.0 at a wavelength of 550 nm was prepared.
  • Example 2 Effect of cocoa bean extract on adhesion of periodontal bacteria to animal cells (1) Preparation of cocoa bean extract-containing medium CBP-containing BHI-YHK medium and HP-PW-containing BHI-YHK medium According to the procedure described in (1), the final medium concentrations of CBP and HP-PW are 0.001, 0.01, and 0.1 mg / mL, respectively, and BBP and HP-PW are used as control groups. A BHI-YHK medium without any addition was prepared.
  • CLPr polyphenol concentration 71% by mass
  • a CLPr-containing 50% ethanol solution was prepared according to the procedure described in Example 1 (1), and then the CLPr-containing BHI-YHK in which the final medium concentrations of CLPR were 0.001, 0.01, and 0.1 mg / mL, respectively.
  • a medium was prepared.
  • Periodontal disease bacteria were described in Example 1 (2) except that they were Porphyromonas gingivalis bacteria solution (10 9 CFU / mL) having an absorbance of 0.3 at a wavelength of 550 nm. Prepared according to procedure.
  • Human gingival epithelial cells were used as animal cells. Human gingival epithelial cells were cultured in a 24-well plate using 1 mL of KGM medium (manufactured by LONZA) for 48 hours under conditions of 5% CO 2 and 37 ° C. CBP-containing BHI-YHK medium, HP-PW-containing BHI-YHK medium and CLPr-containing BHI-YHK medium prepared in (1) above were added to the human gingival epithelial cells after culture at 0.0001 mg / mL, 0.001 mg / mL.
  • KGM medium manufactured by LONZA
  • Example 3 Effect of unfermented cocoa bean extract on periodontal disease bacteria (1) Preparation of unfermented cocoa bean extract-containing medium CBP-containing BHI-YHK medium and HP-PW-containing BHI-YHK medium were prepared in Example 1 (1 ) And BHI-YHK medium with CBP and HP-PW final medium concentrations of 0.01, 0.1, 1.0, and 10 mg / mL, respectively, and CBP and HP-PW as a control group. A BHI-YHK medium without any addition was prepared.
  • Example 4 Effect of unfermented cocoa bean extract on biofilm (1) Preparation of unfermented cocoa bean extract-containing medium CBP-containing BHI-YHK medium and HP-PW-containing BHI-YHK medium were prepared in Example 1 (1 ), The final medium concentrations of CBP and HP-PW are 0.1, 1.0 and 10 mg / mL, respectively, and neither CBP nor HP-PW is added as a control group. BHI-YHK medium was prepared.
  • Biofilm formation A round cover glass is placed in a 24-well microplate to which 1 mL of BHI-YHK medium has been added, and this is inoculated with 10 ⁇ L of the Porphyromonas gingivalis bacterial solution prepared in Example 1 (2). And cultured at 37 ° C. for 16.5 hours. After culturing, the floating bacteria in the well were removed with phosphate buffer (pH 7.2), replaced with fresh BHI-YHK medium, cultured at 37 ° C. for 24 hours, and Porphyromonas gingivalis on the cover glass. A biofilm was formed.
  • the CBP-containing BHI-YHK medium and the HP-PW-containing BHI-YHK medium obtained in (1) above were added to the formed biofilm and allowed to act at room temperature for 1 hour under anaerobic conditions, followed by LIVE / DEAD
  • the biofilm was stained using (trade name) BacLight (trade name) Bacterial aVaibility Kit (manufactured by Thermo Fisher Scientific) and observed with a fluorescence microscope (BZ-X700, KEYENCE). Since viable bacteria are labeled in green and dead bacteria are labeled in red, both can be distinguished under microscopic observation.
  • the fluorescence microscope photograph was image-analyzed and the ratio (ratio of viable bacteria) of the green area with respect to the whole photograph area was computed. Note that green corresponds to a portion shown in white or gray in FIG. 5, and red corresponds to a portion shown in dark gray to black in FIG.
  • Group I a control group
  • Group II Porphyromonas gingivalis-infected group
  • Group III an unfermented cocoa bean extract intake group
  • IV group unfermented cacao bean extract intake and Porphyromonas gingivalis infected group
  • CBP carboxymethylcellulose solution
  • Group II is a Porphyromonas gingivalis solution (10 9 CFU / mL) prepared with 5% CMC, Was administered.
  • Group III administered CBP solution prepared with 5% CMC to a final concentration of 10 mg / mL into the oral cavity of rats treated with Group I
  • group IV a CBP solution prepared with 5% CMC so as to have a final concentration of 10 mg / mL was administered into the oral cavity of the rat treated with group II. All rats were allowed free access to food and drinking water and were housed in an environment of a temperature 23 ° C., humidity 60% and light / dark 12 hour cycle.
  • the body weight was measured immediately after Porphyromonas gingivalis infection (4 weeks old), 1 week after Porphyromonas gingivalis infection (5 weeks old) and at the time of sacrifice (11 weeks old), and the average value of the weight of each group Asked.
  • all rats were sacrificed by decapitation blood under ether anesthesia. This animal infection experiment was conducted with the approval of the Kanagawa Dental University Experimental Animal Ethics Committee.
  • the alveolar bone resorption was measured by measuring the distance from the cement enamel border of the maxillary molar portion of the rat slaughtered in (1) above to the alveolar bone crest at seven locations. Specifically, the skull was heated at 2 atm for 10 minutes, immersed in a 3% sodium hypochlorite solution to remove soft tissue, and then the alveolar bone was stained and dried with a 1% methylene blue solution as a sample. The measurement was performed at a magnification of 40 times under a stereomicroscope.
  • the average value of the measured values at 7 locations was defined as the bone resorption amount per individual, and the average value of 6 animals in each group was expressed in millimeters as the bone resorption amount of each experimental group to obtain the standard deviation (SD).
  • SD standard deviation
  • PCR reagent GoTaq (trade name) Green Master Mix Gene (manufactured by Promega) was used, and gene amplification was performed using an iCycler (manufactured by Bio-Rad).
  • iCycler manufactured by Bio-Rad
  • a DNA sample after completion of the PCR reaction and Smart Ladder (manufactured by Wako Pure Chemical Industries, Ltd.) as a molecular weight marker were subjected to 1.5% agarose gel electrophoresis, followed by photography to detect amplified DNA fragments.

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  • Epidemiology (AREA)
  • Biotechnology (AREA)
  • Medical Informatics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Oncology (AREA)
  • Communicable Diseases (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Birds (AREA)
  • Nutrition Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Cosmetics (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)

Abstract

Le but de la présente invention est de fournir une nouvelle composition destinée à être utilisée dans la cavité buccale, en particulier un rince-bouche qui est efficace pour inhiber l'adhérence de bactéries de maladie parodontale à des cellules animales et supprimer l'expression de pilus de bactéries de maladie parodontale. La présente invention concerne une composition destinée à être utilisée dans la cavité buccale (rince-bouche) qui comprend un extrait de fèves de cacao. Ce rince-bouche peut être utilisé en tant qu'agent pour inhiber l'adhérence de bactéries de maladie parodontale à des cellules animales, un agent pour supprimer l'expression de pilus de bactéries de maladie parodontale, etc.
PCT/JP2018/009638 2017-03-14 2018-03-13 Rince-bouche Ceased WO2018168821A1 (fr)

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JP2017048818A JP6918529B2 (ja) 2017-03-14 2017-03-14 口腔用組成物
JP2017-048818 2017-03-14

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WO2018168821A1 true WO2018168821A1 (fr) 2018-09-20

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4529767A1 (fr) * 2023-09-26 2025-04-02 Berkem Developpement Utilisation d'un extrait de cacao pour la défense des plantes contre des champignons phytopathogènes

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH05944A (ja) * 1991-06-21 1993-01-08 Taiyo Kagaku Co Ltd 歯周病原因菌付着阻害用組成物
JP2000256345A (ja) * 1999-03-02 2000-09-19 T Hasegawa Co Ltd ポリフェノール類の製造方法
WO2003099304A1 (fr) * 2002-05-27 2003-12-04 Morinaga & Co., Ltd. Composition contre des bacteries parodontales, ainsi qu'aliments, boissons et eaux dentaires contenant cette composition contre des bacteries parodontales
JP2006034291A (ja) * 1996-09-06 2006-02-09 Mars Inc ココア成分、高められたポリフェノール含有量を有する食用製品、その製造法、および医学的利用法
JP2010053062A (ja) * 2008-08-28 2010-03-11 Sunstar Inc 口腔用組成物
JP2016504279A (ja) * 2012-11-09 2016-02-12 シャオウェイ・シー テアフラビン抽出物を含むオーラルケア組成物

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH05944A (ja) * 1991-06-21 1993-01-08 Taiyo Kagaku Co Ltd 歯周病原因菌付着阻害用組成物
JP2006034291A (ja) * 1996-09-06 2006-02-09 Mars Inc ココア成分、高められたポリフェノール含有量を有する食用製品、その製造法、および医学的利用法
JP2010187704A (ja) * 1996-09-06 2010-09-02 Mars Inc カカオ成分、高められたポリフェノール含有量を有する食用製品、その製造法、および医学的利用法
JP2000256345A (ja) * 1999-03-02 2000-09-19 T Hasegawa Co Ltd ポリフェノール類の製造方法
WO2003099304A1 (fr) * 2002-05-27 2003-12-04 Morinaga & Co., Ltd. Composition contre des bacteries parodontales, ainsi qu'aliments, boissons et eaux dentaires contenant cette composition contre des bacteries parodontales
JP2010053062A (ja) * 2008-08-28 2010-03-11 Sunstar Inc 口腔用組成物
JP2016504279A (ja) * 2012-11-09 2016-02-12 シャオウェイ・シー テアフラビン抽出物を含むオーラルケア組成物

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4529767A1 (fr) * 2023-09-26 2025-04-02 Berkem Developpement Utilisation d'un extrait de cacao pour la défense des plantes contre des champignons phytopathogènes

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