DK169714B1 - Oily Preparation and Method of Preparation - Google Patents

Description

in DK 169714 B1

OIL PREPARATION AND PROCEDURE FOR PREPARING IT

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BACKGROUND OF THE INVENTION

1. Field of the Invention The present invention relates to an oily preparation and a process for its preparation, and in particular to an oily preparation containing, as effective components, substances which suppress the formation of reactive oxygen and lipid peroxide in the human body. In the following, these substances will be referred to as antioxidant agents.

2. State of the art 20 Recently, attention has been drawn to antioxidant agents which exhibit excellent effects when used to prevent and treat various inflammations and difficult to cure diseases such as chronic arthritis rheumatism, thrombophlebitis, progressive systemic sclerosis, Buerger's disease, Raynaud's disease, difficult-to-treat dermatoulcus, and the like, caused by cellular organization destruction due to reactive oxygen and lipid peroxide, formed in the human body.

It has been clarified in a study conducted by the present inventors that the above antioxidant substances comprise low molecular weight substances such as flavones, polyphenols, tannins, tocopherols, vitamin B 2 and the like contained in a plant growth, and by taking them orally as 35 medicaments. or as a health food, the formation of the above reactive oxygen and lipid peroxide in the body is suppressed (see DK 169714 B1 2

Journal of Japan Pharmacist Association, No. 39, Volume 12, Supplement, "Bioavailability of SOD (Superoxide Dismutase) and Crude Drug", pp. 1097-1119, published December 1, 1987).

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In the past, preparations containing such antioxidants, namely a plant foodstuff according to Japanese patent no. 1366268 and an antioxidant composition disclosed in Niwa's Japanese Untreated Publicly Available Patent Application No. 63-79834 have been known.

The plant food according to No. 1366268 can be obtained by mixing green tea powder with baked unpolished rice powder and soybean powder; by adding a small amount of koi mushroom 15 to the mixture; then immersing this powder mixture in a mixture of sesame oil and soybean oil for about 4 days to extract effective components; by removing precipitates by centrifugation; and by enclosing the residual oily substance in a capsule made of gelatin or the like.

The antioxidant composition of no. 63-79834 is obtained by gentle heating of plant seeds or germs thereof without causing stinging; by subsequently brewing these heated plant materials by the addition of a microorganism; and by adding a vegetable oil from a plant which was also mildly heated. Optionally, vitamin C, vitamin C derivatives or a plant growth containing these are added to the composition.

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As plant seeds, seeds containing antioxidant s such as flavones, polyphenols, tannins, tocopherols, vitamin B1 and the like are used, including rice, wheat, barley, soybean, adzuki bean, corn, hatomugi (barley 35 grains), peas. As a vegetable oil, sesame oil, soybean oil, oil from cotton seed, corn oil, color-time oil, pri- DK 169714 B1 3 mula oil, rice bran oil, rapeseed oil, olive oil and the like are used.

However, the plant feed according to No. 1366268 is poor in content of antioxidant activity, due to its treatment method of the raw plant material.

The antioxidant composition of Niwa No. 63-79834 removed such a problem by remarkably increasing the activity of antioxidant substances by gentle heating of the raw plant material without causing long infrared irradiation, thus counteracting inactivation of high temperature antioxidants by releasing and converting the raw material into the low molecular weight effective components, and by brewing the heat-treated material in this way, to effect further release and conversion into the low molecular weight effective substances.

However, according to the study by the present inventors, it has been made clear since then that although the above antioxidant substances significantly suppress the formation of the reactive oxygen and lipid peroxide in test tubes, the antioxidant activity is not sufficiently demonstrated in the human body.

It is believed that in order to allow antioxidant substances to enter cells suffering from a disease such as an inflammatory reaction or the like due to reactive oxygen and lipid peroxidase, it is necessary to pass through the cell membrane that forms the surface of the cells. However, the human cell membrane is rich in oil and fat components and, therefore, has such a property that only oily substances are allowed to pass, so that the above antioxidant composition having a low content of oily substances has little ability to to pass through the cell-35 limb membrane and penetrate the interior of the cell.

Therefore, the present inventors have repeated studies to improve the ability of the antioxidant composition to penetrate the cell membrane, and at the same time made further improvements in the choice of plant species as raw material and in the treatment method thereof. As a result, an oily preparation has been obtained in which the activity of the antioxidant substances as effective components is high and the penetration ability into the interior of the cell suffering from a disease is high, and thus the present invention is complete. ended.

DISCLOSURE OF THE INVENTION

It is an object of the present invention to provide an oily preparation and a process for their preparation in which the activity of antioxidant substances as effective components is high and where the penetration of penetration into the interior of the cell in a disease location is high.

The process for preparing the oily preparation of the present invention comprises: heating rice germ and / or wheat germ and soybean at a temperature not exceeding 100 ° C; adding koji to the heated material; brew mixture; then pulverizing the mixture; and adding the obtained powdered mixture to an oil mixture comprising an oil obtained from sesame heated at a temperature not exceeding 100 ° C and an oil obtained from crude sesame wherein the ratio of the oil mixture to a total amount of the pulverized mixture and the oil mixture is 60 to 95% by weight.

Because the raw material contains rich antioxidant substances, plant species described in Japanese Un-35 examined Patent Application Laid-open no. Can be used in addition to the mentioned rice germ, wheat germ and soybean. 63-79834, that is barley, adzuki bean, corn, hatomugi (barley), peas and DK 169714 B1 5. However, according to the study conducted by the present inventors, rice germ, wheat germ and soybean are the most preferred raw materials.

5 Along with rice germ, wheat germ and soybean can be listed rice bran, hatomugi (barley), and wheat. Therefore, as a raw material, at least one of rice bran, hatomugi (barley) and wheat can be included together with rice bran and / or wheat bran. In all cases, the ratio of the germ (rice germ and / or wheat germ) in the cereal raw material is preferably at least more than or equal to 1% by weight.

The antioxidant substances in the above cereal raw material form a complicated macromolecular polymer with other substances and therefore have no activity as is, thus it is necessary to break the macromolecular bonds by mild heating or the like, thus releasing low molecular weight antioxidants. However, if the heating temperature is too high, the low molecular weight antioxidant substances are inactivated and it is therefore necessary to select a heating condition that takes this fact into account.

In order to satisfy such a condition, it is necessary to slowly heat the grain feedstock at a temperature not exceeding 100 ° C and to heat for a sufficient period of time. In more detail, the grain feedstock is placed in a vessel made of ceramic material such as earthenware or the like, irradiated with long infrared rays, preferably with 30 wavelengths of 4 to 14 µm, and heating is done while slowly stirring and the temperature is maintained at about 90 to 96 ° C.

The heating time varies depending on the type of grain,

35 so that it cannot be arbitrarily defined. However, it is preferably about 30 minutes to 3 hours. IN

Furthermore, the heating method is not limited to the above method, provided that the antioxidant substances in the feedstock are sufficiently released as low molecular weight substances, while inactivation thereof is avoided.

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After heat treatment as described above, koji (aspergillus orizae) is added to the feedstock to brew it. The brewing conditions are preferably at 20 to 36 ° C for about 2 to 6 days. When brewing is carried out using a fermentation tank, 2 to 3 hours is sufficient. This brewing is done to further promote the release and conversion to low molecular weight of the antioxidant substances in the grain feedstock, and through this brewing step the activity of the antioxidant substances is significantly increased compared to raw materials subjected only to heat treatment.

Next, the brewed material is ground to a fine powder. Grinding can be done using a commercially available mill. However, some types of mills generate a high temperature during use and therefore inactivate the antioxidant substances. Therefore, it is advantageous to use those which do not generate a high temperature during use. For example, a stone mill can be used.

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Next, the oil is prepared by mixing in an appropriate ratio an oil recovered from heated sesame (hereinafter referred to as the sesame paste oil) with an oil extracted from raw sesame, and the above fine powder is added to the oil mixture. The sesame paste oil is an oil produced by grinding and pressing raw sesame after it has been heated slowly for a sufficient period of time at a temperature not exceeding 100 ° C. However, the fine solid content formed by the grinding of sesame remains as is, so that the appearance is given a paste state. This sesame paste oil richly contains the low molecular weight antioxidants and by using it it is possible to obtain the oily preparation having a high activity of the antioxidant compound.

However, this sesame paste oil has a high viscosity, and the size of an oil microdroplet is also large, so that the preparation in which only the sesame paste oil is added to the above fine powder has little penetrating ability into the interior of cells in a disease region. However, when seed oil recovered from raw sesame is added to the sesame paste oil, the size of the microdroplet may be small and the penetrating ability into the interior of cells in a disease area is enhanced.

The crude sesame oil is an oil extracted by grinding the raw sesame as it is, squeezing it and removing solid contents, and it is commercially available as a regular sesame oil. Since the mixing ratio of the sesame paste oil to the regular sesame oil is different and also depending on the amount of fine powder to be added, the ratio cannot be uncritically defined. It is preferably 1 to 3 parts by weight of the general sesame oil relative to 1 part by weight of the sesame paste oil.

The ratio of the oil mixture to the fine powder added thereto is such that the mixed oil becomes 60 to 95% by weight relative to a total amount of both. If the oil mixture is less than 60% by weight, the ability to pass through the cell membrane becomes poor. On the other hand, if the oily mixture exceeds 95% by weight, the concentration of antioxidant substances as the effective components becomes small, so that the effect of suppression of the reactive oxygen and lipid peroxide is also diminished.

The oily preparation according to the present invention, orally consumed, can be prepared by adding the fine powder to the oil mixture and enclosing it in a gelatin capsule or the like as soon as the oil mixture is prepared. However, the material mixture can preferably mature at 20 to 35 ° C for about 3 to 30 days, or more advantageously at 28 to 30 ° C for about 1 week before encapsulation.

By applying the ripening treatment, the release ice and the conversion of antioxidant substances to low molecular weight are further promoted due to the koji remaining in the fine powder and the antioxidant substances are well absorbed in the oil mixture so that the activity and the osmotic ability of the reactants into the the interior of the cell is further enhanced.

The oily preparation of the present invention may also include a raw material of cereals comprising rice germ or wheat germ and soybean and optionally at least one of the risk 20 lid, hatomugi (barley) or wheat which is made into a fine powder as is, without heating and brewing. , and added to the oil mixture together with the above heated and brewed fine powder.

25 In the fine powder of the grain raw material which is not heated and brewed, the antioxidant substances as the effective component are not released and converted into low molecular weight substances. However, when added to the oil mixture with the heated and brewed fine powder, the release 30 and the conversion of antioxidant substances to low molecular weight occur gradually even after encapsulation, due to the koji remaining in the brewed fine powder, so that there is such an advantage , that the effect of suppressing the formation of the reactive oxygen and lipid peroxide is maintained for a long period.

DK 169714 B1 On the other hand, in the oil preparation in which only the heated and brewed fine powder is used, due to enzyme reactions and the like, which continue continuously even after encapsulation, the antioxidant substances as the effective components are gradually degraded, so that the effective period is short compared to a preparation in which the untreated fine powder is added.

However, in the case where the amount of untreated fine 10 powder added is too large, the activity of the effective component decreases. Therefore, it is advantageous that the untreated fine powder is about 0.5 to 1 part by weight based on 1 part by weight of the heated and brewed fine powder. Incidentally, in this case too, where 2 15 types of fine powder are included, the oil mixture is 60 to 95% by weight relative to the total amount.

In the thus obtained oily preparation of the present invention, both the activity of the antioxidant and the osmotic force into the interior of the cell is high, and as also clarified from results of clinical studies as described below, it exhibits remarkable effects. against various inflammations and difficult to cure diseases for which the therapeutic effect has been insufficient with conventional anti-inflammatory agents.

The oily preparation of the present invention can be encapsulated with gelatin or the like and taken orally as a medicine. In addition, in the case of abnormal pigmentation such as dermatitis, chloroasma, freckles and the like, or wrinkles and the like, the oily preparation may be applied directly to an affected area. In addition to dermatitis and the like, this oily preparation can also be used for advanced and curable diseases such as chronic arthritis, thromboembolism, phlebitis, progressive systemic scleroderma, Buerger's disease, Raynaud's disease, difficult-to-treat dermatoulcus and the like. The preparation according to the invention also has effects on the treatment and prevention of other various pollution-related diseases, burns, external wounds, fatigue, hangovers, constipation and the like.

In addition, the oily preparation of the present invention has no side effects due to the use of only cereals, koji and sesame as raw materials, so that it is possible to ingest orally as a health food to maintain and improve health. Needless to say, when preparing a preparation, it is optional to add additional medications or ingredients useful for the whole variety, such as various vitamins, minerals and the like, and flavors, flavors, dyes and the like.

Example 20 Preparation Each portion of rice germ, soybean, rice bran, hatomugi (barley) and wheat was transferred to a vessel made of ceramic material irradiated with long infrared rays with a wavelength of 4 to 14 µm, and heated to 90 to 96 ° C for 3 hours. hours of slow stirring so as not to cause stinging.

Next, koji were added as 3% of a total amount to brew at 36 to 40 ° C for 72 hours, and then the brewed material was ground to a fine powder using a stone mill. Further, the above-mentioned five grain raw materials, which were not heated and brewed, were painted with a stone mill 30 to obtain a fine powder in the same amount. On the other hand, sesame was heated in the same way as above grain materials, and after it was painted with a stone mill, it was pressed to get a sesame paste oil.

Next, 28.1 parts by weight of the sesame paste oil were mixed with 48.9 parts by weight of a commercially available sesame oil; to this mixture were added 11.5 parts by weight of the untreated fine powder; and the resulting mixture was stirred homogeneously. The resulting mixture was matured at 28 ° C for 1 week and then encapsulated in gelatin to obtain an oily preparation.

Effect of the Invention [I] In Vitro Test 10 (1) As an in vitro test of the ability to penetrate and penetrate oily sites, the oily preparation of the Example Example was added to a TBA (thiobarbituric acid) reaction system in which an oily unsaturated fatty acid (decosahexaenoic acid) was reacted with 15 reactive oxygen, produced by ultraviolet rays, and formed lipid peroxide, and the degree of suppression of lipid peroxide formation was measured.

To 0.1 cc of decosahexaenoic acid diluted 100-fold was added 1.8 mg / ml of the sample of the oily preparation of the invention and the lipid peroxide formed was measured with the TBA reaction. In the TBA reaction, 0.2 ml of 7% sodium dodecyl sulfate, 2 ml of 0.1 N HCl, and 0.3 ml of phosphorus tungstic acid were mixed; 1.8 mg / ml of the sample was added to the mixture; 1 ml of a reagent was added in which 0.67% TBA was mixed with acetic acid in a 1: 1 ratio; and the measurement was performed with estimates at 515 nm and emission at 553 nm using a fluorescence spectrophotometer.

As comparative examples, the plant nutrient was used according to Japanese patent no. 1366268 (Comparative Example 1) and the antioxidant composition disclosed in Japanese Unpublished Public Application Patent Application No. 63-79834 (Comparative Example 2), and measurements were made in the same manner. Results are shown in Table 1.

12 DK 169714 B1

Table 1 5 Sample Average (6 minute value)

Control 461 ± 62 10 (UV +) · £ · jfe tfe

Oily preparation according to 121 ± 13 of the present invention (1.8 mg / ml)

Ich

Comparative Example 1 271 ± 34 (1.8 mg / ml) *

Comparative Example 2 358 ± 45 (1.8 mg / ml) 20 UV +: ultraviolet irradiation λ · 0.01 <P <0.05 (relative to control): P <0.01 (relative to control) 25: P <0.0001 (relative to to control)

Test results 30

Although each of the samples significantly suppressed the formation of lipid peroxide (TBA reactants) from the unsaturated fatty acid (decosahexaenoic acid) with irradiation of ultraviolet rays (2), the oily preparation of the present invention particularly repressed this formation (P <0.0001). . This result supports the fact that the oily preparation of the present invention has a higher activity of the antioxidant fat, and a greater penetration ability into the interior of the cell in which a disease occurs, as compared to conventional preparations. (Comparative Examples 1 and 2).

(2) An [3,4-H₂J antioxidant substance was prepared in which the low molecular weight antioxidant substance contained in the oily preparation of the present invention was labeled with an isotope (H); it was introduced into a human tissue 3 in a test tube; and the count numbers (cpm) of [H] bound to its cell membrane were measured with a scintillation counter; thereby, the ability to penetrate the cell membrane was tested (labeled tritiated thymidine was 2 C / mM).

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The same studies were also performed for Comparative Examples 1 and 2. Results are shown in Table 2. As shown in Table 2, the oily preparation of the present invention had the highest affinity for cell membranes in 20 relative to Comparative Examples 1 and 2.

Table 2 5 14 DK 169714 B1

Sample cpm built-in

Control 15643 cpm / 10 6 cells 3 Oily preparation according to 56372 cpm / 10 cells of the present invention (1.8 mg / ml) 3

Comparative Example 1 33451 cpm / 10 cells (1.8 mg / ml) 3

Comparative Example 2 30567 cpm / 10 cells (1.8 mg / ml) 20 (3) The oily preparation of the present invention was subjected to ultrasonic treatment and added to a reactive oxygen (neutrophil and xanthine xanthine oxidase) system so that a converted concentration in the living tissue was 1.6 mg / ml; and three types of reactive oxygen f ^ 2 ^ 2) were measured to compare with the case without addition (control). The converted concentration in living tissue is an amount believed to be present in blood when a normal intake amount per day (930 g in the case of the oily preparation of the present invention) is absorbed into a living body.

The measurement methods for the three types of reactive oxygen are as follows.

DK 169714 B1 15

For O 2 ~, the method was used in which the amount of ferric cytochrome C reduction due to O 2 was measured at a wavelength of 550 nm with a Beckman spectrophotometer and converted to the amount of O 2 -.

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For Η2θ2 ,. Depending on the fact that H 2 O 2 decreases the fluorescence produced by scopoletin in the presence of peroxidase, using scopoletin and peroxidase, the degree of fluorescence reduction was measured using a Hitachi Ltd. fluorescence spectrophotometer. with impact at 370 nm and emission at 460 nm.

For OH * using the principle that alpha-ketomethiol-butanoic acid (KMB) reacts with OH * to form ethylene gas, the method was used in which ethylene gas was quantitated by a gas chromatograph from Hitachi Ltd., for which method of converting the formed ethylene gas to OH *. The same studies were also performed for Comparative Examples 1 and 2. Results are shown in Table 3.

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Table 3 16 DK 169714 B1

Reactive Oxygen 5

Sample 02 "H202 OH '

Control 1,532 nmol 485 pmol 854 pmol 10 Oily Preparation 0.589 nmol 161 pmol 283 pmol according to the present invention (1.8 mg / ml)

Comparative Example 1 1.285 nmol 403 pmol 707 pmol 15 (1.8 mg / ml)

Comparative Example 2 0.231 nmol 538 pmol 108 pmol (1.8 mg / ml) 20

The antioxidant effect of the oily preparation of the present invention was slightly less than that of Comparative Example 2, however, the antioxidant effect shown was greater than that of Comparative Example 1. However, as shown in Tables 1 and 2 above, Comparative Example 2 a lower content of oily substances so that it has little ability to penetrate the cell in which a disease occurs.

Therefore, when considered as a whole, the oily preparation of the present invention in which the antioxidant effect is high and the penetration ability into the interior of the cell in a disease area is highest is most excellent as an antioxidant agent.

DK 169714 B1 17 II Clinical studies

The therapeutic efficacy was investigated in 96 patients with 5 autoimmune diseases, collagen diseases such as chronic arthro-rheumatism, hemastenosis, nephritis, hapatocirrhosis, chloasma, freckles and the like, which have hitherto been resistant to or worsened after treatment with any of the -steroidal antiphlogistic drugs, steroids and the antioxidant composition of Comparative Example 2. The results are shown in Table 4.

18 DK 169714 Pg

Table 4 Sample 5 Oily Comparative Comparative Preparation Example 1 Example 2 of Near- (1.8 mg / ml) (1.8 mg / ml) Present Invention 10 (1.8 mg / ml)

Disease

Chronic 13/18 (72%) 0/7 (0%) 4/14 (28%) arthroreum matism

Angitis 3/5 (60%) 0/3 (0%) 1/4 (25%)

Progressive 9/12 (75%) 2/10 (20%) 6/12 (50%) 20 Systemic scleroderma

Dermato- 6/8 (75%) 2/12 (16%) 6/13 (46%) 25 Myocitis

Thrombosis 2/3 (66%) 0/4 (0%) 1/5 (20%) phlebitis 30 Buerger's 6/8 (75%) 0/3 (0%) 3/6 (50%) disease

Raynaud's 10/12 (83%) 3/8 (37%) 7/12 (58%) disease 35 varices 8/11 (72%) 4/9 (44%) 11/15 (73%) cruris

Difficult 3/5 (60%) 1/5 (20%) 2/6 (33%) 40 curable dermatoulcus

Chloasma 11/14 (78%) 4/12 (33%) 17/25 (68%) freckles 45 50

Claims (7)

An oily preparation, characterized by being obtained by heating rice germ and / or wheat germ and soybean at a temperature not exceeding 100 ° C, by making the heated material into a fine powder, by adding koji to the powdered material and brewing the powdered material, and by adding the material thus brewed to an oil mixture containing a pasta oil recovered from sesame heated at a temperature not exceeding 100 ° C, and an oil recovered from raw sesame, wherein a ratio of said oil mixture is obtained to a total amount of the fine powder and the oil mixture is 60 to 95% by weight. An oily preparation according to claim 1, characterized in that it further comprises a grain raw material containing rice germ and / or wheat germ and soybean made into a fine powder without heating and brewing. An oily preparation according to claim 1 or 2, characterized in that said heated or unheated cereal raw material comprises, in addition to rice germ and / or wheat germ and soybean, at least one of rice bran, hato-mugi (barley) or wheat. An oily preparation according to claim 1 or 2, 30, characterized in that it is encapsulated for oral ingestion as a medicine or health food. Process for preparing an oily preparation according to claim 1, 2 or 3, characterized in that the heated cereal raw material is heated by long infrared rays having a wavelength of 4 to 14 µ1 μπι. Process for preparing an oily preparation according to claim 1, 2 or 3, characterized in that the cereal raw material is brewed at 20 to 36 ° C for 2 to 6 days after heat treatment. Process for preparing an oily preparation according to claim 1, 2 or 3, characterized in that the fine powder is added to the oil mixture so that it matures at 20 to 35 ° C for 3 to 30 days.