JP3308497B2 - SOD-like extract composition extracted from coffee beans - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a SOD-like extract composition extracted from coffee beans. More specifically, the present invention relates to various diseases caused by active oxygen, for example, diseases caused by impaired blood flow (myocardial infarction, stroke, hypertension, menstrual cramps,
Stiff shoulders, neuralgia, back pain, hangovers, etc., adult diseases / medical diseases (cancer, nephritis, hepatitis, diabetes, etc.), beauty and skin diseases (stains, freckles, rough skin, chills, constipation, wrinkles, atopic dermatitis, etc.) That can be widely used for treatment, improvement, etc.
D-like extract composition.

[0002]

2. Description of the Related Art Superoxide dismutase (hereinafter, referred to as superoxide dismutase)
It is called SOD. ) Catalyzes the disproportionation reaction of the superoxide radical (O ₂ ⁻ ) generated by one-electron reduction of oxygen molecules (the following formula) at a rate close to the diffusion rate, and reduces the intracellular O ₂ ⁻ concentration. It is an enzyme. Reactive oxygen species typified by 2O ₂ ⁻ + 2H ⁺ → H ₂ O ₂ + O ₂ O ₂ ⁻ are usually produced from activated phagocytic cells such as macrophages in vivo, and exhibit bactericidal and tumoricidal effects. . However, it is known that these reactive oxygen species do not have selective toxicity and can act on normal cells, resulting in various obstacles to living organisms. For example, membrane damage due to lipid peroxidation, protein structural change due to oxidative modification of protein, DNA
It has been shown that as a result of cleavage and the like, cells show a damaging effect and cause various diseases.

[0003] Therefore, O ₂ ^- is the removal enzyme of SOD,
It exists to protect the living body from reactive oxygen species,
In view of the fact that it is effective against diseases and the like that are considered to be caused by this reactive oxygen species, its reaction mechanism, physiological mechanism and the like have been studied in recent years.
Molecular Mechanism of Elimination / Action ”(2nd Edition, published by Kyoritsu Shuppan Co., Ltd., edited by Minoru Nakano et al., Pp. 223-230)]. Also,
There is a fact that SOD activity is low in cancer cells.
Although the direct causal relationship between SOD and carcinogenesis is not clear,
There is also a report that injection of OD or SOD-like substance into cancer cells suppresses proliferation (p. 64).

[0004] Green coffee beans contain various components such as lipids, carbohydrates, proteins and caffeine in addition to chlorogenic acids, which are tannin components. In recent years, coffee has been shown to have various effects such as anti-mutagenicity and antioxidation.
For example, stabilization of vitamin C by chlorogenic acids (JP-A-6-9603), prevention of deterioration of natural fragrances (JP-A-4-345693), browning inhibitor for pigments (JP-A-5-32909, Tokuhei 1) 22877), prevention of deterioration of flavor (Japanese Unexamined Patent Publication No. 6-38723), a gastric mucosa protective agent (Japanese Patent Publication No. 63-502349),
N β _{-Alkanoyl-5-hydroxytryp}
There are known an antioxidant characterized by containing tammine (Japanese Patent Application Laid-Open No. 8-157816), a cosmetic composition containing coffee tree seed extract containing chlorogenic acid as an essential component (Japanese Patent Application Laid-Open No. 8-92057), and the like. .

[0005]

A safe and SOD action (including not only an action to reduce the concentration of active oxygen, but also an action to prevent and improve various diseases which may be caused by the action). It is very useful for human health and beauty if there is one that has it and is used for foods and the like, and the need for it is extremely large. There is a lot of stress as in recent years,
Since the necessity is particularly large in a situation where many diseases caused by the disease have occurred, development of a product having a safe and excellent SOD action has been strongly desired.

[0006] On the other hand, many recent studies on the physiological activity of coffee often involve extracting a specific component from coffee beans and examining the physiological activity. However, as described above, since the components in coffee beans contain many substances, it is difficult to separate and identify all of them and to examine their biological activities. In addition, there may be a case where not only a specific component but also a plurality of components have a synergistic effect. However, at present, knowledge on such a synergistic effect has not yet been sufficiently obtained.

[0007] Furthermore, even if a substance that has been shown to be effective in vitro is orally administered to a living body, the digestive and absorption processes in various parts of the digestive system are involved in a complicated manner, so that the same effect is not necessarily achieved. Not exclusively. Since and in the above-mentioned application examples are used in vitro, all are merely findings on in vitro effects, and nothing is described about the effects when actually administered to a living body. In addition, although the above-mentioned application example is used for the human body, it is a cosmetic used for the skin, and therefore, nothing is described about the pharmacokinetics and effect when actually applied to a living body. Further, in the above application example, chlorogenic acid is administered in vivo, but its effect is mainly a gastric mucosal protective effect by an astringent effect, that is, it binds to a protein on the gastric mucosa to form an insoluble precipitate, It forms a coating and protects the gastric mucosa. Therefore, in this case, it is considered that the administered chlorogenic acid is hardly absorbed from the digestive system, and the components in the coffee beans are absorbed from the digestive system,
It is unknown what effect it has on the whole body.

The present invention has been made in view of the above-mentioned circumstances, and uses a coffee bean as a raw material, has a safe and excellent SOD action in a living body, and is particularly effective even in a trace amount. The purpose is to provide.

[0009]

In view of the above, the present inventors have conducted various studies focusing on the physiological activity of the complex synergistic action of various components in coffee beans. Of the extracted extract, even in trace amounts in vivo
The present invention has been shown to have an OD action, and the present invention has been completed.

The SOD-like extract composition of the first invention comprises water,
Hot water, ethanol, ethyl acetate, or ethanol
Is a solvent mixture of ethyl acetate and water or hot water
And the extract extracted from coffee beans
Hea bean extract. " ) In an amount of 0.0003 to 0.5% by weight, and has an SOD action in vivo. As the above-mentioned "coffee beans", for example, any varieties such as Arabica, Robusta, and Rebellica may be used, and the place of production is not particularly limited. In addition, coffee beans produced for coffee drinks in the cultivation area are sorted and removed in a large amount when the coffee beans are sorted for roasting. Can be used as "beans". The “coffee bean extract” may be a liquid obtained by filtering the extract, a liquid obtained by subjecting the extract to post-treatment such as decolorization, or a concentrated liquid obtained by concentrating the extract. Incidentally, a powder obtained by spray-drying this extract may be used.

The extraction method for obtaining the above-mentioned "coffee bean extract" is not particularly limited. The raw coffee beans may be unground or ground, and may be subjected to pretreatment such as removal of impurities as long as the quality of the extract can be maintained. The extraction solvent may be water or hot water, or an organic solvent such as ethanol or ethyl acetate, or a mixed solvent of these organic solvents with water or hot water. Since the present extract is water-soluble, among these solvents, water (including hot water) or a mixed solvent of ethanol / water is preferable. The extraction conditions are not particularly limited, but usually, room temperature or heat extraction is preferred. Regarding the heating temperature and the heating time, various conditions can be set as long as the extraction can be sufficiently performed and the quality of the extract can be maintained. Further, in the above-mentioned extraction, substances that assist the extraction can be added as long as the quality of the extract can be maintained. For example, a reducing substance such as L-ascorbic acid may be added in addition to enzymes such as protease and cellulase. The pH of the extract usually shows 5.0 to 6.0 when L-ascorbic acids and the like are added, but there is no limit to this pH as long as the extract does not deteriorate in quality.

When the coffee bean extract concentration in the SOD-like extract composition of the first invention exceeds 0.5% by weight,
Since the SOD effect tends to reach a plateau and the SOD effect increases particularly at 0.005 to 0.5% by weight, the coffee beans in the SOD-like extract composition of the first invention are Extract concentration is 0.000
It is used in a proportion of 3 to 0.5% by weight, preferably 0.005 to 0.5% by weight.

[0013] The SOD-like extract composition of the present invention contains 0.0003 to 0.5% by weight of a coffee bean extract and has an SOD activity ratio of 1. measured by a nitro blue tetrazolium (hereinafter referred to as NBT) reduction method. It is a 1 to 3.0
Can be .

The "NBT reduction method" refers to the reaction of xanthine with oxygen in the presence of xanthine oxidase, and
₂ ^- In the reaction process that produces the diformazan by reducing NBT, a substance showing a SOD activity is present,
Results to reduce the concentration of, ^- which O ₂ ^- react with, O ₂
Since the amount of diformazan produced decreases, the degree of the decrease is determined as the inhibition rate, and the SOD in the sample is determined.
This is a method to determine the activity. The “SOD activity ratio” is
Based on the test method described in the following example, a sample containing no coffee bean extract was orally administered to mice, and SOD measured by an NBT reduction method from the obtained blood sample.
It means the ratio between the activity and the SOD activity measured by the NBT reduction method from a blood sample obtained by administering a sample containing a coffee bean extract at a predetermined concentration to a mouse.

[0015] The SOD-like extract composition of the present invention can have an SOD activity ratio of 1.1 to 3.0 as measured by the NBT reduction method. In particular, the coffee bean extract concentration in the SOD-like extract composition is 0.0003-0.0
Even at a concentration as low as 5% by weight, the SOD activity ratio measured by the NBT reduction method can exhibit a value of 1.1 to 1.7. The coffee bean extract of the first invention is excellent in SOD action in a living body, and thus can be used as a therapeutic composition for treating a disease caused by active oxygen. Further, as shown in the second invention, it can be used as a water-soluble beverage or a food additive composition.

[0016]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be specifically described below with reference to examples. (1) Production of SOD-Like Extraction Composition 100 kg of coffee beans (type: Java, produced in Indonesia) is pulverized by an interactive pulverizer (manufactured by Mutual Sangyo Co., Ltd.), and the pulverized beans are passed through a magnetic force of 10,000 Gauss to remove metallic foreign substances I do. Next, 5 times the amount of water was added to the ground beans, and 0.02% of citric acid and 0.15% of L-ascorbic acid were added with stirring to adjust the pH to 5.6 to 5.6.
When the mixture is dissolved, stop stirring and leave it overnight without heating to extract. Then, solids are removed from the extract with a centrifuge. Subsequently, the extract is transferred to a tank, mixed with activated carbon in an amount of 0.06% based on the theoretical amount of ground beans, heated to 50 ° C., and stirred for 30 minutes to be decolorized. Then, as a pre-coat 0.5-1.5kg in the filter
Diatomaceous earth was used as a body coat, and about 1.8 kg of diatomaceous earth was used as a body extract for filtration to produce a coffee bean extract.

After the above coffee bean extract was spray-dried to obtain a powder, the powder was dissolved in distilled water to prepare a SOD-like extract composition having the following concentrations as a test sample. Test sample of test group 1: 0.0005% (W / V) aqueous solution Test sample of test group 2: 0.005% (W / V) aqueous solution Test sample of test group 3: 0.05% (W / V) aqueous solution Test sample of test group 4: 0.5% (W / V) aqueous solution Test sample of test group 5: 5.0% (W / V) aqueous solution

(2) Test method As test animals, 6-week-old Wistar rats (5-week-old rats purchased from Japan SLC, Inc.) were used as males.
0 and 60 females were used. After pre-breeding the Wistar rats for 7 days, the rats were divided into 20 groups (10 males and 10 females) each, and the rats were divided into 6 groups (control group and test groups 1 to 5). And room temperature 23 ± 1 ° C., humidity 45-55%, 12
The animals were reared for 28 days under the conditions of time illumination (8:00 to 20:00). During the test period, 1.0 ml / day / rat of the test sample was orally administered to the control group and each test group using a gastric probe. The same amount of distilled water was orally administered to the control group. Oriental Yeast Kogyo Co., Ltd. "Oriental MF" was used as a feed for free intake, and distilled water was freely supplied. On the last day of the test, blood was collected from the heart, and the SOD activity and lipid peroxide level in the serum were measured.

(3) Measurement of SOD activity and lipid peroxide content From the blood sample obtained as described above, SOD activity and lipid peroxide content were measured by the following measurement methods. SOD
The activity was determined by measuring the activity of the kit using "SOD Test Wako" (manufactured by Wako Pure Chemical Industries, Inc., an in vitro diagnostic drug, approval number (63AM)).
No. 0285) by the above-mentioned NBT reduction method. The results are shown in Table 1, FIG. 1 and FIG. Note that "S
As described above, the “OD activity ratio” refers to the S value of the control group.
The ratio of the SOD activity of each test group to the OD activity.

[0021] Lipid peroxide is available as a measurement kit "Lipid peroxide-Test Wako (Fluorescent Yagi method)" manufactured by Wako Pure Chemical Industries.
(In Vitro Diagnostic Drug, Approval Number (61AM) No. 4641
No.) and the thiobarbituric acid (hereinafter referred to as TBA) method. This method converts lipid peroxide to T
The concentration of lipid peroxide is determined by measuring the fluorescence intensity of a red substance produced by heating BA with acidic conditions. The results are shown in Table 2, FIG. 2 and FIG. The “lipid peroxide concentration ratio” is the ratio of the lipid peroxide concentration of each test group to the lipid peroxide concentration of the control group.

(4) Test Results From Table 1, the SOD activity was higher in all test groups than in the control group, and a significant difference (P <0.01) was observed. The composition was shown to significantly enhance the ability of the body to scavenge active oxygen. From the graph of FIG. 1, the coffee bean extract concentration was 0.005 to 0.5.
It can be seen that the percentage by weight of the SOD activity ratio is large when the weight% is used. On the other hand, it has been shown that when the coffee bean extract concentration exceeds 0.5% by weight, the increase in SOD activity reaches a plateau, so that there is an upper limit of the coffee bean extract concentration suitable for increasing SOD activity. You can see that Further, FIG.
It can be seen from the graph that a sharp increase in the SOD activity ratio is observed when the coffee bean extract concentration is 0.0005% by weight, as compared to 0.005% by weight and 0.05% by weight. That is, it is one of the great features of the extract composition according to the present invention that the SOD activity is significantly increased as compared with the control group even if the extract composition according to the present invention is in a very small amount.

[0022] Lipid peroxide is an index of a disorder of active oxygen disorder, and increases when its degree is severe. From Table 2, as a result of the measurement, it was confirmed that the amount of lipid peroxide in each of the test groups was significantly lower than that in the control group (test groups 1 to 5: P <0.05). FIG.
According to the graph, the coffee bean extract concentration was 0.005 to
It can be seen that at 0.5% by weight, the rate of inhibition of lipid oxidation is large. On the other hand, the coffee bean extract concentration was 0.5% by weight.
When the ratio exceeds, the tendency of lipid oxidation suppression to slow was observed. This indicates that there is an upper limit of the coffee bean extract concentration that is suitable for suppressing lipid oxidation, as in the case of the above-mentioned SOD activity. Furthermore, from the graph of FIG. 4, when the coffee bean extract concentration is 0.0005% by weight, compared with 0.005% by weight and 0.05% by weight,
It can be seen that the lipid peroxide concentration is rapidly decreasing, that is, even when the extract is in a very small amount, the suppression of lipid oxidation as compared to the control group is the same as in the case of the above SOD activity. One of the major features of the extraction composition according to the present invention is that it has a great advantage.

From the above results, it can be seen that the ingestion of the extract composition according to the present invention increased the SOD activity in the body and suppressed the oxidation of lipids. Also,
No harmful components and heavy metals are found in any of the components of the extraction composition according to the present invention. In particular, the extraction composition according to the present invention is safe because it is made from a completely natural product. And, as shown in the above results, since the extract composition according to the present invention is excellent in SOD action, S
It is considered to have an effect due to the OD action. In addition, the coffee bean extract contained in the extract composition according to the present invention is water-soluble, and can be used as an aqueous beverage, food, etc. by dissolving in water as shown in the above test, and It can be said that it is excellent in absorption.

The present invention is not limited to the specific embodiments described above, but may be variously modified within the scope of the present invention according to the purpose and application. That is,
The form of the extract is usually a liquid such as an aqueous solution or an undiluted solution as shown in the above examples, but is not limited to this. A powder product obtained by impregnating the extract with an absorbent powder, a granulated product Tablets, microcapsules, and the like, into which other powder components such as granules, extenders, and the like are blended can be used. Further, these aqueous solutions, powdered products and the like are filled in a predetermined container in a commercial form, or used alone or mixed with other agents (whether aqueous solution, oily liquid or powder). There is no particular limitation as to whether or not to do so. For example, it may be a potion type, may be filled in a container of another shape, or may be a product in which a powdery product is filled in a stick-shaped container (bag). Furthermore, it may be blended and dispersed in conventional soft drinks, drinks, dairy products, oil products, and the like. In addition, this dispersion does not ask whether it is a water-in-oil type or an oil-in-water type. Further, other nutrient components (for example, various vitamins, calcium ion components, iron ion components, etc.), medicinal components, seasoning components, odor components, and the like may be blended. Of these, water-soluble components are particularly preferred. This is because the product can be uniformly dissolved.

[0025]

The extract composition according to the present invention is excellent in the SOD action in a living body, and therefore is effective for the effects (for example, blood pressure suppressing effect) caused by the SOD action. Therefore, the extract composition according to the present invention provides various diseases caused by reactive oxygen species, for example, diseases due to impaired blood flow (myocardial infarction, stroke, hypertension, menstrual pain, stiff shoulder, neuralgia, back pain,
Hangover), treatment and improvement of adult diseases and medical diseases (cancer, nephritis, hepatitis, diabetes, etc.), beauty and skin diseases (stains, freckles, rough skin, coldness, constipation, wrinkles, atopic dermatitis, etc.) It is considered that it exerts an excellent effect.

Further, the coffee bean extract contained in the extract composition of the present invention is water-soluble, so that it has an excellent absorption effect and can be easily ingested by oral administration.
Since the SOD action is exhibited even in a very small amount, it is not necessary to add a large amount. As a result, it can be considered to be used in a wide range of fields such as water-soluble beverages and foods. Furthermore, since the extraction composition according to the present invention uses completely natural products as raw materials, it is safe without harmful components and heavy metals.

[Brief description of the drawings]

FIG. 1 is a semilogarithmic graph showing the relationship between the concentration of a coffee bean extract and the SOD activity ratio.

FIG. 2 is a semilogarithmic graph showing a relationship between a concentration of a coffee bean extract and a lipid peroxide concentration ratio.

FIG. 3 is a graph showing the relationship between the coffee bean extract concentration and the SOD activity ratio when the coffee bean extract concentration is in the range of 0 to 0.005% by weight.

FIG. 4 is a graph showing the relationship between the coffee bean extract concentration and the lipid peroxide concentration ratio when the coffee bean extract concentration is in the range of 0 to 0.005% by weight.

[Table 1]

[Table 2]

──────────────────────────────────────────────────続 き Continuation of the front page (56) References JP-A-10-120696 (JP, A) (58) Fields investigated (Int. Cl. ⁷ , DB name) A23L 1/30 A61K 35/78 JICST file ( JOIS)

Claims (2)

(57) [Claims] An extract extracted from coffee beans using water, hot water, ethanol, ethyl acetate, or a mixed solvent of ethanol or ethyl acetate and water or hot water as an extraction solvent, and extracting an extract from coffee beans in an amount of 0.0003 to 0.5. An SOD-like extract composition, which has an SOD effect by weight. 2. An extract extracted from coffee beans using water, hot water, ethanol, ethyl acetate, or a mixed solvent of ethanol or ethyl acetate and water or hot water as an extraction solvent, wherein the extract extracted from coffee beans is 0.0003 to 0.5. wherein wt% water-soluble beverage or food additive composition characterized by <br/> have a SOD activity
Thing .