US20030096029A1 - Remedies for allergic diseases and process for producing the same - Google Patents

Remedies for allergic diseases and process for producing the same Download PDF

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US20030096029A1
US20030096029A1 US10/169,874 US16987402A US2003096029A1 US 20030096029 A1 US20030096029 A1 US 20030096029A1 US 16987402 A US16987402 A US 16987402A US 2003096029 A1 US2003096029 A1 US 2003096029A1
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remedy
allergic diseases
sugar
fermentation
producing
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Harui Fukuda
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Priority to US10/389,430 priority Critical patent/US7048933B2/en
Publication of US20030096029A1 publication Critical patent/US20030096029A1/en
Priority to US10/674,752 priority patent/US20040062825A1/en
Priority to US11/327,720 priority patent/US20060269632A1/en
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/145—Fungal isolates
    • 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
    • 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
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • 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
    • A23L33/14—Yeasts or derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7016—Disaccharides, e.g. lactose, lactulose
    • 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
    • A61K36/13—Coniferophyta (gymnosperms)
    • A61K36/15—Pinaceae (Pine family), e.g. pine or cedar
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/06—Antiasthmatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/04—Antipruritics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A61P27/14—Decongestants or antiallergics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/16—Otologicals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
    • C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00—Technologies for the production of fuel of non-fossil origin
    • Y02E50/10—Biofuels, e.g. bio-diesel

Definitions

  • the present invention relates to a remedy for allergic diseases, for example, asthma, atopic dermatitis, conjunctivitis, rhinitis and food allergy, and to a method of producing the same. Further, the present invention relates to yeast isolated from the above-mentioned remedy.
  • a human body has an immune system which is a defense mechanism which, when extraneous substances such as bacteria and viruses invade body, antagonizes them and protects body. Allergies are caused due to excess action of this immune system. Recently, increasing number of people are suffering from allergies possibly because of, though the details are not clear, air pollution, change of dietary life, physical or mental stress increase, environmental changes such as room pollution and the like due to change in resident circumstances, or change in human body constitution.
  • the allergic diseases include asthma, atopic dermatitis, allergic conjunctivitis, allergic rhinitis such as pollenosis, food allergy and the like.
  • the first remedy for these allergic diseases is to avoid the allergens, though the remedy is rather passive.
  • the house When dusts or mites in a house are the allergens, the house must be cleaned to remove the allergens, and when a pollen is the allergen, in the season of the pollen floatation, going out should be held or a mask should be worn to prevent suction of the allergen, and in the case of a food allergy, the food causing the allergy should not be eaten.
  • a passive remedy is troublesome to the patient and significantly restricts the activities of the patient.
  • atopic dermatitis causes distresses not only to the patient but also to the parents of the patient. Though the most of the patients are cured before maturity, there is a case where the dermatitis lasts to the adulthood. In such a case, adolescent men and women are distressed by thickening of the skin of face, breast, inside of elbow and knee and by severe itching. As the remedy for this, topical treatments are the major methods to treat the patient, and depending on the symptom, adrenal cortical hormones, antihistamine drugs and other anti-inflammatory agents are used. When itching is strong, systemic application of an antihistamine agent would be necessary.
  • the object of the present invention is to provide a remedy which can completely cure allergies by administrating the remedy in a short period of time without causing side effects, and a method of producing such a remedy.
  • the remedy for allergic diseases (allergies) of the present invention can be obtained by mixing shoots of plant belonging to the family Pinaceae with water and saccharides followed by spontaneous fermentation, and the remedy is effective for treating allergic diseases such as asthma, atopic dermatitis and the like.
  • the above-mentioned plant belonging to the family Pinaceae plant belonging to the genus Pinus is preferable, and as the above-mentioned saccharides, sugar is preferable.
  • the remedy for allergic diseases of the present invention can be obtained by mixing shoots of pine leaves with water and sugar followed by spontaneous fermentation, and the remedy is effective for treating allergic diseases such as asthma, atopic dermatitis and the like.
  • the present invention relates to a method of producing the above-mentioned remedy for allergic diseases
  • the method of producing the remedy for allergic diseases comprises (1) a step of dissolving saccharides in sterilized water to prepare a saccharides solution, and (2) a step of adding shoots of plant belonging to the family Pinaceae to said saccharides solution followed by spontaneous fermentation.
  • the above-mentioned plant belonging to the family Pinaceae a plant belonging to the genus Pinus is preferable, and as the above-mentioned saccharides, sugar is preferable.
  • the method of producing a remedy for allergic diseases of the present invention comprises (1) a step of dissolving sugar in sterilized water to prepare a solution containing dissolved sugar, and (2) a step of adding shoots of pine leaves to said solution containing dissolved sugar followed by spontaneous fermentation.
  • the above-mentioned spontaneous fermentation is preferably conducted under anaerobic conditions at 10 to 70° C., preferably 20 to 60° C. for about 3 to 9 months, preferably 4 to 8 months.
  • the more specific method of producing a remedy for allergic diseases comprises (1) a step of dissolving sugar in hot water to prepare a sugar aqueous solution then the solution being cooled to ambient temperature, and (2) a step of adding water-washed shoots of pine leaves to said sugar aqueous solution, placing the solution into a vessel and sealing the vessel followed by spontaneous fermentation, and the spontaneous fermentation is preferably conducted by placing the sealed vessel at a place receiving direct sunlight until early winter.
  • the present invention also contemplates a health food obtained by mixing shoots of plant belonging to the family Pinaceae with water and saccharides followed by spontaneous fermentation, and the health food is preferably obtained by mixing shoots of pine leaves with water and sugar followed by spontaneous fermentation. Further, the present invention encompasses a fermentation products obtained by the spontaneous fermentation set forth above, namely, a novel yeast isolated from the remedy for allergies.
  • the yeast has been deposited as HARUISAN A-3 with Independent Administrative Agency, National Institute of Advanced Industrial Science and Technology (AIST), International Patent Organism Depositary, Chuo No.
  • Higashi 1-1-1, Tsukuba City, Ibaraki Prefecture old name: National Institute of Bioscience and Human-Technology National institute of Advanced Industrial Science and Technology, Higashi 1-1-3, Tsukuba City, Ibaraki Prefecture, name has been changed on Apr. 1, 2001) on Mar. 12, 2001, and specified by deposit No. FERM BP-7499.
  • a series of yeasts having the microbiological properties equivalent to the deposited yeast are also included in the present invention.
  • a fermentation product obtained by the fermentation using the above-mentioned yeast is included, and as the fermentation product, pharmaceutical preparations such as allergic curative medicines, health foods, health drinks, and raw materials of cosmetics, and the like are exemplified.
  • the health food and health drink indicate foods and drinks such as supplements and the like used for the purpose of improving body constitution and maintaining health.
  • FIG. 1 is a microscopic photograph (differential interference, X 2400) showing an example of the ascospore of isolated yeast isolated from the remedy for allergies (fermentation product) of the present invention.
  • the medium malt extract agar medium was used.
  • FIG. 2 shows the results of the mutagenicity screening tests made on a remedy for allergies of the present invention, where base pair-substituted type strains are used (TA100: ⁇ , TA1535: ⁇ , WP2uvrA: ⁇ ).
  • A represents a result in the case of utilizing no metabolism activation ( ⁇ S9)
  • B represents a result in the case of utilizing metabolism activation (+S9), respectively.
  • FIG. 3 is a graph showing the results of the mutagenicity screening test of on a remedy for allergies of the present invention, and shows the results obtained by using a frame shift type strain (TA98: ⁇ , TA1537: ⁇ ).
  • TA98 frame shift type strain
  • A represents a result in the case of utilizing no metabolism activation ( ⁇ S9)
  • B represents a result in the case of utilizing metabolism activation (+S9), respectively.
  • FIG. 4 is a graph showing the drug effect of a remedy for allergies of the present invention.
  • the contraction ratio of an extracted trachea from a guinea pig sensitized with ovalbumin (OVA) is plotted against the dose of OVA when a remedy for allergies of the present invention is applied to the organ bath.
  • OVA ovalbumin
  • “A” represents a result in group I
  • “B” represents a result in group II.
  • ⁇ represents control (injection water)
  • * represent a result in the case of addition of 0.02% of a remedy for allergies of the present invention.
  • * represents p ⁇ 0.05, significant as compared with the control.
  • FIG. 5 is a graph showing the drug effect of a remedy for allergies of the present invention, and shows the result of an ear swelling test (ear edema thickness measuring test).
  • DNCB 1-chloro-2,4-dinitrobenzene
  • HAR a remedy for allergies of the present invention
  • injection water distilled water: DW
  • HAR acetone-olive oil mixture
  • * * represents p ⁇ 0.01, significant for group III
  • # represents p ⁇ 0.05, significant for group II.
  • the remedy for allergies of the present invention is obtained by mixing shoots of plant belonging to the family Pinaceae with water and saccharides followed by spontaneous fermentation.
  • G Don Larix gmelini (Rupr.) Kuzeneva, Larix Kaempferi (Lamb.) Carriere, Picea abies (L.) Karst., Picea glehnii (Friedr. Schmidt) M. T. Masters, Picea jezoensis (Sieb. & Zucc.) Carriere var. hondoensis, Picea jezoensis (Sieb. & Zucc.) Carriere, Picea koyamae Shirasawa, Picea polita (Sieb. & Zucc.) Carriere, Pinus x densi-thunbergii Uyeki, Pinus densiflora Sieb.
  • sucrose invert sugar, maltose and the like are exemplified.
  • sucrose is preferable from the standpoint of easy availability, and as the sucrose used, any sugar such as white sugar, black sugar, yellow soft sugar, beet sugar, millet sugar and the like can be used, and white sugar is preferable.
  • water can be used, previously sterilized water is preferably used to prevent proliferation of saprophytic bacteria, and as the sterilization method, any known methods generally employed to sterilize water can be used, for example, water can be sterilized by boiling and the like.
  • the remedy for allergies of the present invention can be obtained by dissolving saccharides in the above-mentioned sterilized water to prepare a saccharides aqueous solution, adding to the saccharides aqueous solution a shoots of plant belonging to the family Pinaceae, and subjecting the mixture to spontaneous fermentation.
  • the shoots of plant belonging to the family Pinaceae to be added to said aqueous solution may be a shoots collected from any kind of plants belonging to the family Pinaceae, and particularly, a shoots collected from a plant belonging to the genus Pinus is preferable.
  • the preferable season for collecting the shoots is the season after completion of blooming of the plant, and the shoots collected in this season has the highest effectiveness as a remedy for allergies and therefore is preferable.
  • the shoots at the peak of the branch are preferably collected and used when blooming of these flowers is completed.
  • the starting solution used in the fermentation For preparing the starting solution used in the fermentation, about 0.5 kg of saccharide is dissolved in 1 liter of water. Then, to the resulting solution, about 25 shoots of plant belonging to the family Pinaceae are added. In this case, the saccharide is not required to be completely dissolved and there is no problem if the saccharide presents remaining undissolved in the solution.
  • the starting solution to be fermented may advantageously contain shoots of plants belonging to the family Pinaceae such as shoots of pine leaves in the ratio set forth in the above, and there is no problem if it contains leaves and flowers other than the shoots.
  • the spontaneous fermentation can be conducted under anaerobic conditions and the fermentation is effected by allowing the starting solution to stand still at 10 to 70° C., preferably 20 to 60° C. for 3 to 9 months, preferably 4 to 8 months.
  • the starting solution is filled into a light-shielded vessel and the like and the vessel is sealed air tightly.
  • the fermentation is realized by placing this sealed vessel at a place receiving direct sunlight until around early winter.
  • the vessel is opened, and the solid materials such as shoots of plant belonging to the family Pinaceae are removed to obtain a remedy for allergies of the present invention.
  • the above-mentioned early winter is determined in consideration of the shoots collecting period, namely, charging period, and if the blooming period is from early April to early May and if the shoots collection period after completion of blooming is from middle May to early June in the producing area, the period of fermentation completion shall be early winter (middle November), however, this is only one example, and this period can optionally be changed.
  • the remedy for allergies of the present invention is effective for allergic diseases, particularly, asthma and atopic dermatitis, and in view of the mode of the action, the remedy is believed to be effective for improvement and treatment of, in addition to the above-mentioned diseases, type I allergic diseases such as anaphylactic shock by drugs, allergic rhinitis, acute urticaria, pollenosis, food allergy and the like, type II allergic diseases such as autoimmune hemolytic anemia, haemolysis and thrombocytopenia due to drug allergy, type III allergic diseases such as serum diseases, SEL, acute nephritis, polyarteritis and the like, type IV allergic diseases such as contact dermatitis, allergic encephalitis and the like.
  • type I allergic diseases such as anaphylactic shock by drugs, allergic rhinitis, acute urticaria, pollenosis, food allergy and the like
  • type II allergic diseases such as autoimmune hemolytic anemia, haemolysis and thrombocytopenia due to drug allergy
  • the remedy for allergies of the present invention uses a fermentation product per se obtained by the spontaneous fermentation, however, a sweetener and flavoring agent may be added to the product to improve the acceptance of the product in drinking, or various additives such as a preservative and the like may be added for storing the product for a long period of time or for other purposes.
  • the remedy for allergies of the present invention in the case of adult, in general, about 30 to 50 ml of the remedy is administered twice or three times a day. In the case of child, half dosage of that in the adult administration may be used. Since the remedy for allergies of the present invention has no toxicity and no mutagenicity and therefore is safe, there is no problem if an amount over the above-mentioned dosage is used.
  • the fermentation product of the present invention obtained by the spontaneous fermentation can be used not only as a remedy for allergies but also as health food or health drink.
  • the health food or health drink are not used mainly for remedy, but used to improve body constitution and maintain healthy condition.
  • the present invention includes yeast isolated from the above-mentioned fermentation product. This yeast is isolated as described below.
  • a fermentation product which is the remedy for allergies of the present invention was used as the specimen.
  • GPLP agar plate culture method was used to culture the microorganisms on the plate, and the colonies dominantly grown on the culture plate was picked up to obtain the isolated yeast.
  • the morphological and physiological observations were performed, and the yeast was identified by consulting literatures (Kurtzm an, C. P. et at., “The yeasts, ATaxonomic Study” 4-th edition (1998), Elsevier Science B. V.; Barnett, J. A. et al., “Yeasts: Characteristics and identification”, 3-rd edition (2000), Cambridge University Press, these literatures are incorporated as parts of the specification of the instant application by reference).
  • the total number of yeast in the fermentation product was 1.4 ⁇ 10 5 /g.
  • the results form the morphological and physiological observations on the isolated yeast are shown in Table 1 and an example of ascospore of the isolated yeast is shown in FIG. 1.
  • the isolated yeast is identified as Zygosaccharomyces bisporus from the morphological and physiological properties.
  • the Zygosaccharomyces bisporus belongs to ascomycetous yeast, mates between the individual cells and forms 1 to 4 ascospores of sphere to ellipse form. Further, Zygosaccharomyces bisporus is osmosis resistant yeast and isolated from fermentation foods, soft drinks and the like.
  • yeast of the present invention is identified as Zygosaccharomyces bisporus from the morphological and physiological properties, DNA sequence itself is close to Zygosaccharomyces bailii rather than Zygosaccharomyces bisporus . Accordingly, it is recognized that the yeast of the present invention is a novel strain differing from both type strains. The applicant named this novel yeast as “HARUISAN A-3” which is deduced to belong to Zygosaccharomyces genus. Presently, it is not clear whether this novel yeast “HARUISAN A-3” is just a novel strain or is a novel species or genus.
  • the isolated novel yeast of the present invention is believed to significantly contribute to effectiveness of the remedy for allergies together with shoots of plants belonging to the family Pinaceae, and therefore is extremely useful yeast.
  • the remedy for allergies of the present invention can be produced, for example, by the following method.
  • White sugar was added and dissolved into hot water, and cooled to around room temperature, and the resulting sugar water and water-washed shoots of pine leaves were placed in a vessel, for example, a plastic vessel, the vessel is sealed, and the vessel is placed at a place receiving direct sunlight until early winter (around middle November, in Fukushima pref.) to cause spontaneous fermentation for the production.
  • the vessel was opened around early winter, and the pine leaves were removed to obtain the remedy for allergies of the present invention.
  • yeast was present in the order of 10 2 to 10 5 /g in the fermentation product, though the yeast concentration was slightly irregular depending on the lot of the fermentation product.
  • the yeast existed in the above fermentation products corresponded to the deposited novel yeast.
  • the oral toxicity of the remedy for allergies of the present invention was examined.
  • the stock solution of the remedy for allergies of the present invention and that obtained by adding honey (10 wt %) to the stock solution were used as the specimens.
  • Two test groups each consisting of 5 male and 5 female SD [Crj: CD(SD)IGS] rats were administered the stock solution or the honey added stock solution at a rate of 2000 mg/kg and the control group was administered the injection water (dose: 0 mg/kg) alone.
  • the specimens and the injection water were forcibly administered to the rats orally by using a disposable syringe (volume: 1 mL) equipped with a per os stomach conductor.
  • the toxic symptom and approximate lethal dose were investigated over the period of 15 days after the administration (including administration day).
  • the mutagenicity of the remedy for allergies of the present invention was investigated.
  • the remedy for allergies was applied to the histidine-dependent Salmonella typhimurium, TA98, TA100, TA1535 and TA1537 strains, and tryptophan-dependent Escherichia coli,WP2uvrA strain according to the revised plate method of Ames et al. (Maron, D. M. et al., “Revised methods for the Salmonella mutagenicity test”, Mutation Res., Vol. 113, pp. 173 to 215 (1983), this literature is incorporated as parts of the specification of the instant application by reference), and the mutagenicity was investigated under the presence of the metabolism activation or the absence of the metabolism activation.
  • FIGS. 2 and 3 show the result in the base pair-substituted type strain (TA100: ⁇ , TA1535: ⁇ , WP2uvrA: ⁇ ).
  • FIG. 3 shows the results in the frame shift type strain (TA98: ⁇ , TA1537: ⁇ ).
  • “A” represents the result in the absence of the metabolism activation
  • “B” represents the result in the presence of the metabolism activation to which S9 mix was added.
  • As the negative control substance distilled injection water that was used as the solvent to prepare the specimen was administered.
  • AF-2 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide
  • 2-Aminoanthracene (2-AA) 2-Aminoanthracene (2-AA)
  • SA sodium azide
  • 9-Aminoacridine (9-AA) 9-AA
  • AF-2, 9AA and 2-AA were dissolved in DMSO and SA was dissolved in distilled injection water, respectively, and used depending on the strain and on the presence or absence of the metabolism activation.
  • Ovalbumin (OVA) used as an allergy inducing substance was dissolved in physiological saline.
  • OVA and Freund's complete adjuvant (FCA) were mixed at equivalent amount, to give an administration specimen.
  • OVA or physiological saline for sensitization was administered subcutaneously twice a week for two weeks, 4 time in total.
  • the specimen i.e., the remedy for allergies (fermentation product) of the present invention or injection water was orally administered to the animals for 3 days, i.e., preceding 3 days before the day of tracheal extraction (isolation).
  • Trachea was extracted from one animal from each group over 5 days from 11 day after the final sensitization administration, and a smooth muscle contraction test was conducted. Further, in extracting trachea, abdominal section was carried out and the blood was collected from inferior vena cava to obtain the serum, and the serum anti-OVA IgG antibody value was measured to confirm the establishment of sensitization, and an influence by oral administration of the above-mentioned remedy for allergies of the present invention was also investigated.
  • a micro titre plate (NUNC) on which OVA had been fixed was blocked with Block Ace (manufactured by Dainippon Pharmaceutical Co., Ltd.) and the blocked plate was used for the measurement.
  • OVA-induced contraction in group II showed a suppression tendency by applying the remedy for allergies of the present invention to the organ bath (concentration in bath: 0.02%) (when OVA 10 ng/mL, p ⁇ 0.05, see FIG. 4).
  • the remedy for allergies of the present invention showed from no influence to suppression tendency in all groups (for example, in group III, when histamine 10 ⁇ 6 M, p ⁇ 0.05).
  • the mean absorbance+3 SD (standard deviation) in group IV was 0.243.
  • the mean absorbance of 10 4 -fold diluted sera in group I and group II conducted the sensitization-administration of OVA were 1.465 and 1.488, respectively, confirming the accomplishment of OVA sensitization.
  • the remedy for allergies of the present invention suppresses the liberation of inflammatory substances such as histamine through the stabilizing action on the plasma membrane and the like to show the anti-allergic action.
  • the remedy for allergies of the present invention was administered at morning and night, twice a day, each in an amount of about 30 to 50 mL. After around one month administration, the symptom became lighter than before, and around about three months, the symptom became very well.
  • the remedy for allergies of the present invention was administered at morning and night, twice a day, each in an amount of about 30 to 50 mL, then, after about two weeks administration, the symptom was improved in both females.
  • the remedy for allergies of the present invention was administered twice to three times a day, each in an amount of about 30 to 50 mL.
  • the inflammation on the rear surface of knee was remedied and original skin became visible, and 3 weeks after, the wound at neck disappeared, then, the inflammation at the left arm was remedied, and two months after, the inflammation at the right arm was also remedied, beautiful skin could be recovered.
  • the remedy for allergies of the present invention can completely cure allergies, particularly, asthma and atopic dermatitis, by administration in a short period of time without causing side effects as compared with conventional allergic remedies.
  • the present invention provides an extremely useful remedy for allergies.

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US10/674,752 US20040062825A1 (en) 2000-06-14 2003-09-30 Remedies for allergic diseases and process for producing the same
US11/327,720 US20060269632A1 (en) 2000-06-14 2006-01-05 Remedies for allergic diseases and process for producing the same

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US10/674,752 Abandoned US20040062825A1 (en) 2000-06-14 2003-09-30 Remedies for allergic diseases and process for producing the same
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KR101252548B1 (ko) * 2006-01-19 2013-04-08 (주)아모레퍼시픽 해송자 추출물을 유효성분으로 함유하는 화장료 조성물
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KR20030031000A (ko) 2003-04-18
KR100539627B1 (ko) 2005-12-28
CA2412652A1 (en) 2002-12-12
US20060269632A1 (en) 2006-11-30
AU2001274526A1 (en) 2001-12-24
EP1297837A1 (en) 2003-04-02
US20030175258A1 (en) 2003-09-18
JP3471789B2 (ja) 2003-12-02
US20040062825A1 (en) 2004-04-01
KR20050072157A (ko) 2005-07-08
US7048933B2 (en) 2006-05-23

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