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US 9,308,229 B2Granted
Method of inhibiting angiogenesis
AssigneesHiroko Itoh, Powerful Healthy Food Corporation, Sun Chlorella Corp.
InventorsHitoshi Ito, Hiroko Itoh, Masaki Fujishima, Yukari Arakawa, Fukuyoshi Nakada
Life of this patent
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Abstract
A method of inhibiting angiogenesis by applying a kind of protein polysaccharide of a (1→6)-β-D-glucan obtained from a residue of extraction of crushed dried fruiting body of Agaricus blazei Murill (himematsutake) with 80% ethanol aqueous solution and hot water at 100 degrees C.
Claims8
A method of inhibiting angiogenesis by applying a substance obtained by the steps of:
- extracting a fruiting body of Agaricus blazei Murill (himematsutake) with a lower aliphatic alcohol or a lower aliphatic alcohol containing not more than 20% of water to obtain Residue S,
- extracting Residue S with hot water to obtain Residue I,
- extracting Residue I with 1% ammonium oxalate aqueous solution to obtain Residue II,
- extracting Residue II with 5% NaOH aqueous solution containing 0.05% NaBH4 and filtering to obtain Filtrate I,
- neutralizing Filtrate I with acetic acid to pH of 5 to 6 to obtain Supernatant I,
- adding ethanol to Supernatant I to obtain Precipitate I,
- discarding compounds having molecular weight lower than 10,000 from Precipitate I and drying to obtain Fraction I,
- extracting Fraction I with 80% formic acid aqueous solution and filtering to obtain Filtrate II,
- fractionating Filtrate II by CHCl3-n-butanol (5:1, v/v) to CHCl3-n-butanol layer and Aqueous layer I,
- extracting Aqueous layer I with 5% lithium chloride-dimethylacetamide aqueous solution to obtain Precipitate II, and
- further fractionating Precipitate II by ethanol precipitation.
8 more elements
A method of inhibiting angiogenesis by applying a substance obtained by the steps of:
- extracting a crushed dried fruiting body of Agaricus blazei Murill (himematsutake) with 80% ethanol aqueous solution to obtain Residue S,
- extracting Residue S with hot water at 100 degrees C. to obtain Residue I,
- extracting Residue I with 1% ammonium oxalate aqueous solution at 100 degrees C. to obtain Residue II,
- extracting Residue II with 5% NaOH aqueous solution containing 0.05% NaBH4 at 30 degrees C. and filtering to obtain Filtrate I,
- neutralizing Filtrate I with acetic acid to pH of 5 to 6 to obtain Supernatant I,
- adding a five times volume of ethanol to Supernatant I to obtain Precipitate I,
- dialyzing Precipitate I to discard compounds having molecular weight lower than 10,000 and lyophilizing to obtain Fraction I,
- extracting Fraction I with 80% formic acid aqueous solution at 85 degree C. and filtering to obtain Filtrate II,
- fractionating Filtrate II by CHCl3-n-butanol (5:1, v/v) to CHCl3-n-butanol layer and Aqueous layer I,
- extracting Aqueous layer I with 5% lithium chloride-dimethylacetamide aqueous solution at 30 degrees C. to obtain Precipitate II, and
- further fractionating Precipitate II by ethanol precipitation.
8 more elements
Dependencies are inferred from the claim text.