Drug – bio-affecting and body treating compositions – Plant material or plant extract of undetermined constitution... – Containing or obtained from leguminosae
Reexamination Certificate
2000-04-24
2001-10-09
Naff, David M. (Department: 1651)
Drug, bio-affecting and body treating compositions
Plant material or plant extract of undetermined constitution...
Containing or obtained from leguminosae
C435S183000, C435S200000
Reexamination Certificate
active
06299911
ABSTRACT:
TECHNICAL FIELD
The present invention relates to an &agr;-glucosidase inhibitor containing Touchi as an active ingredient, which can be used in pharmaceuticals, food products, health foods, particular health care foods and the like.
BACKGROUND OF THE INVENTION
It is described that an &agr;-glucosidase inhibitor inhibits an &agr;-glucosidase which is localized at a fine villus in a small intestine, and controls a rapid increase in blood sugar after meal and next increase in insulin level (Diabate Medicine, 10, 688 (1993)). Since it suppresses metabolism of a carbohydrate (especially oligosaccharide derived from starch, sucrose and the like) also in human and the other animals, and exhibits an inhibitory effect of blood sugar increase, it is effective in improving a hyperglycemic condition as well as various diseases induced by a hyperglycemia such as obesity and diabetes. And a food product obtained by adding &agr;-glucosidase thereto is a suitable meal for a patient having a symptom and also for a healthy human on the purpose of prophylaxis against such symptom.
As an &agr;-glucosidase inhibitor derived from a food, for example, a enzymatic hydrolysate of an animal protein or a vegetable protein were disclosed in Japanese Unexamined Patent Publication No. 9-65836(1997) and tea polyphenol was disclosed in Japanese Unexamined Patent Publication No. 5-17364(1993).
However, the &agr;-glucosidase inhibitor disclosed in Japanese Unexamined Patent Publication No. 9-65836(1997) had a problem that it should be taken in a large amount as a food for exerting its activity. And the &agr;-glucosidase inhibitor disclosed in Japanese Unexamined Patent Publication No. 5-17364(1993) had problems that it was complicated to purify polyphenol and it was necessary to take a large amount in case of routinely consumed tea. An object of the present invention is to provide an excellent &agr;-glucosidase inhibitor, which has a high activity and can readily be ingested.
DISCLOSURE OF THE INVENTION
We made an effort to solve the problems described above and finally discovered that Touchi, which is used as an ingredient of Chinese food, had an excellent &agr;-glucosidase inhibitory activity to complete the present invention.
Namely, the present invention relates to an &agr;-glucosidase inhibitor comprising Touchi as an active ingredient (Claim
1
);
the &agr;-glucosidase inhibitor of Claim
1
, characterized in an extract obtained by extraction of Touchi with water and/or alcohol is used as Touchi (Claim
2
), and
the &agr;-glucosidase inhibitor of Claim
2
, wherein an amount of a component having a molecular weight of at least 3000 in said extract obtained by extraction with water and/or alcohol is at most 20% by weight (Claim
3
).
BEST MODE FOR CARRYING OUT THE INVENTION
Touchi employed in the present invention is usually one obtained by steaming a soybean and fermenting it with koji (Aspergillus). And Touchi includes Daitokuji natto, Hama natto, Tera natto, Shiokara natto, Ikkyuji natto that are fermented soybeans and the like in Japan.
While an ordinary fermentation is performed by preserving in salt, a product similarly produced without salt or a product desalted at completion of the fermentation may also be employed in the present invention.
Touchi is used in an ordinary form, and powder obtained by pulverizing after drying and slurry obtained by pulverizing in water also exhibit a glucosidase inhibitory effect. But extracts obtained by extraction of Touchi with water and/or alcohol are preferable from the viewpoint of exhibiting a potent &agr;-glucosidase inhibitory activity.
Examples of the alcohol employed for extraction are methanol, ethanol, propanol, butanol and the like. Methanol and ethanol are preferable. Examples of the extraction method are stirring extraction, preserving extraction, supercritical extraction and the like. Usually stirring extraction or immersing extraction is employed. When water and alcohol are employed in a combination use, a ratio (weight) of water to alcohol is preferably 1/10 to 10/1, more preferably 1/5 to 5/1.
When the extraction is conducted with water, to Touchi as it is or Touchi obtained by pulverizing is added water in an amount of 1 to 20 times, and Touchi is extracted by stirring extraction at 40 to 60° C. for 12 to 20 hours, or at 100 to 140° C. for 1 to 3 hours.
When the extraction is conducted with alcohol or a combination of water and alcohol, to Touchi as it is or Touchi obtained by pulverizing is added alcohol or a mixture of water and alcohol in an amount of 1 to 15 times, and Touchi is extracted by immersing extraction at 40 to 60° C. for 3 to 10 hours, or at a room temperature for 10 hours to 7 days. If necessary, water and alcohol can be distilled off to adjust a concentration thereof.
An extract obtained after the above extraction step can be used as it is. Alternatively, it may be used in the form of powder after distilling water and alcohol.
The extract usually contains 30 to 90% by weight of the components having a molecular weight of at least 3000, as discussed later.
In order to obtain an effect by ingestion in a small amount, an extract may be subjected to a solvent fractionation using a polar organic solvent such as methanol, ethanol, propanol, butanol, chloroform, ethyl acetate, toluene, hexane or benzene to concentrate an active component, or may be purified by at least one suitable purification method selected from the group consisting of alumina column chromatography, silica gel column chromatography, ion exchange chromatography, hydrophobic chromatography, and high performance liquid chromatography.
In the invention, it is preferable that an amount of the components having a molecular weight of at least 3000 is at most 20% by weight in the extract obtained by water and/or alcohol extraction of Touchi described above from the viewpoint of exhibiting an excellent glucosidase inhibitory activity. Concretely, it is preferable to use a product obtained by removing any components having a molecular weight of at least 3000 from the above extract to be in an amount of at most 20% by weight thereof (hereinafter referred to as “purified product”), more preferably 0.1 to 15% by weight.
The above-mentioned molecular weight of the sample is measured by a gel filtration chromatography. Concretely, using an amino acid, a peptide and a protein [Ala (molecular weight 89), Ile-Tyr (molecular weight 330), Asp-Gly-Leu-Tyr-Pro (molecular weight 563), neurotensin (molecular weight 1637), and ribonuclease (molecular weight 13700) as s standard reference, which have the known molecular weights, an extract is eluted in the following conditions to determine a retention time, and a molecular weight is determined by using a calibration curve obtained by plotting the molecular weights on the ordinate and the retention times on the abscissa logarithmically.
In the invention, an amount “% by weight” of the components having a molecular weight of at least 3000 is represented as “% area” of the peaks corresponding to components having a molecular weight of at least 3000 after dividing the peaks at the position of the molecular weight 3000 on an elution chromatogram.
(Fraction Conditions)
Column: Protein Pak60 (made by Waters) 5×250 mm
Mobile phase: 50% by volume acetonitrile aqueous solution containing
0.1% by volume TFA (trifluoroacetic acid)
Flow rate: 1 mL/min
Detection: RI
Sample amount: 1 mg
TABLE 1
Relationship between molecular weight and elution time
Elution time
Standard substance
Molecular weight
(minutes)
Ala
89
37
Ile—Tyr
330
33
Asp—Gly—Leu—Tyr—Pro
563
31
Neurotensin
1637
26
Ribonuclease
13700
19
In the above-mentioned conditions, since a substance having a molecular weight of 3000 is eluted at 25 minutes, any substance eluted at not more than 25 minutes is regarded as components having a molecular weight of at least 3000.
The followings are examples of a method for removing a component having a molecular weight of at least 3000.
[1] a method for removing fractions having a molecular weight of at least 3000 by means of a gel filtrati
Fujita Hiroyuki
Fukushima Nobuhiro
Kasai Takanori
Kawabata Jun
Armstrong Westerman Hattori McLeland & Naughton LLP
Meller Mike
Naff David M.
The Nippon Synthetic Chemical Industry Co., Ltd.
LandOfFree
Extract of Touchi containing an &agr;-glucosidase inhibitor does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Extract of Touchi containing an &agr;-glucosidase inhibitor, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Extract of Touchi containing an &agr;-glucosidase inhibitor will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2616824