GB2268185A

GB2268185A - Oily preparation and method of production thereof

Info

Publication number: GB2268185A
Application number: GB9312763A
Authority: GB
Inventors: Emiyo Niwa; Kozo Niwa
Original assignee: Individual
Current assignee: Individual
Priority date: 1992-06-22
Filing date: 1993-06-21
Publication date: 1994-01-05
Anticipated expiration: 2013-06-21
Also published as: BE1006911A3; SE9302141L; NO932282L; DK73293A; JPH069421A; ES2049189B1; FR2692442A1; JP2955126B2; AT406935B; ES2049189A1; NL9301083A; SE512781C2; IS4038A; IS1657B; CN1065433C; DE4320526A1; AU4143293A; KR0138738B1; AU656681B2; GB9312763D0

Abstract

An oily preparation and its production method where the activity of antioxidant substances as effective components are high and the penetrating ability into the inside of the cell in 8 disease portion is high, and its production method. The oily preparation is prepared by heating a cereal raw material comprising rice germ and/or wheat germ and soybean at a temperature not exceeding 100(C; making it fine powder after koji is added to the heated material to brew it; and adding thus brewed powder to an oil mixture comprising an oil obtained from sesame heated in the same manner and an oil obtained from raw sesame, wherein a ratio of oil mixture to a total amount of the fine powder and the oil mixture is 60 to 95 % by weight. This oily preparation can be used to prevent and treat various inflammations and inveterate diseases resulting from or worsened by destruction of cellular organization by reactive oxygen and lipid peroxide produced in a human body.

Description

2268185 Oily preparation and method of production thereof

BACKGROUND OF THE INVENTION 1. Field of the Invention

The present invention relates to an oily preparation and its production method, and particularly to an oily preparation containing, as effective components, substances which suppress production of reactive oxygen and lipid peroxide in human bodies. Hereinafter, these substances will be referred to as antioxidant substances. 2. Related Art Statement

Recently, attention has been attracted to antioxidant substances which exhibit excellent effects when used for preventing and treating various inflammations and inveterate diseases such as chronic arthrorheumatism, thrombophlebitis, progressive systemic sclerema, Buerger's disease, Raynaud's disease, intractable dermatoulcer and the like caused by destruction of cellular organization by reactive oxygen and lipid peroxide produced in human bodies.

It has been clarified according to a study by the present inventors that the above-mentioned antioxidant s.dbstances comprise low molecular weight substances such as flavonoids, polyphenols, tannins, 2 tocopherols, vitamin B, and the like contained in a plant body, and by taking -them orally as a medicIne or a health food, the production of above-mentioned reactive oxygen and lipid peroxide in the body Is suppressed (see Journal of Japan Pharmacist Association, No. 39, Vol. 12, supplement, "Bioavailability of SOD (Superoxide Dismutase) and crude drug", pp. 1097-1119, published on December 1, 1987).

Coventionally, as preparations containing such antioxidant substances, a votanical nutrient of Japanese Patent No. 1386268, and an antioxidant composition described in _Kiwa's Japanese Unexamined Patent Application Laid-open No. 63-79834 have been known.

The votanical nutrient of -No. 1366268 is obtained by mixing green tea powder with powder of baked unpolished rice and soybean powder; adding a small amount of koji fungus to the mixture; immersing, then, this powder mixture in a mixture of sesame oil and soybean oil fot about 4 days to extract effective components; removing precipitates by centrifugation; and enclosing remained oily substance in a capsule made of gelatin or the like.

The antioxidant composition of No. 6379834 is obtained by iildly heating plant seeds or germs thereof not to produce scorch; subsequently brewing these heated 3 plant materials by adding a microorganism; and adding thereto a plant oil obtained from a plant which was also mildly heated. Optionally, vitamin C, vitamine C derivatives or a plant body containing them is added to the composition.

As plant seeds, is used a seed which contains antioxidant substances such as above-mentioned flavonolds, polyphenols, ranni-ns, tocopherols, vitamin B, and the like, including rice, wheat, barley, soybean, adzuki bean, corn, hatomuzi (pearl barley), pea. As plant oil, is used sesame oil, soybean oil, cotton seed oil, corn oil, safflower oil, evening primrose oil, rice bran oil, rape oil, olive oil and the like.

However, the votanical nutrient of '-o. 1366268 is low in its antioxidant activity, because of its method of processing the raw plant material.

The antioxidant compositlon of N1wa, No. 6379834 obviated such a problem by remarkably increasing the activity of antioxidant substances by mildly heating the raw plant material not to produce scorch with far- infrared radiation, thus preventing deactivation of antioxidant substances due to high temperature, liberating and converting he raw material to the low molecular weight effective components, and by brewing thus-heat treated material to promote further liberation 4 and conversion to the low molecular weight effective substances.

However, according to the study by the present inventors since then, it has been clarified that -although above-mentioned antioxidant substance remarkably suppresses the production of the reactive oxygen and lipid peroxide in test tube. the antioxidant atvivities are not exhibited In human bodies sufficiently.

It is considered that, in order to allow the antioxidant substances to arrive at the inside of cells suffering a disease such as an inflammatory reaction or the like due to reactive oxygen and lipid peroxide, it is necessary to pass through celi. membranes which cover surfaces of the cells. However, the cell membrane of human Is abundant with oil and fat components, and hence it has such a property that only oily substances are allowed to pass through, so that above-mentioned antioxidant composition having a low content of oily substances has a low ability to pass through the cell membrane to penetrate into the Inside of the cell.

Thus, the present inventors have repeated studies in order to improve the cell membrane passing ability of the antioxidant composition, while making further improvement in selection of plant species as a raw material and its processing method. Consequently, there has been obtained an oily preparation in which the activity of the antioxidant substances as effective components is high and the penetrating ability into the inside of the cell suffering a disease is high, completing the present invention.

DISCLOSURE OF INVENTIOX

It is an object of the present invention to provide an oily preparation and its production method in which the activity of antioxidant substances as effective components is high and the penetrating ability into the inside of the cell at a disease place is high.

Method of producing 'the oily preparation of the present invention compsises: heating rice germ and/or wheat germ, and soybean at a temperature not exceeding 100 0 C; adding kojl into the heared material; brewing the mi-xture; then, powdering the mix-cure; and adding the obtained powde'red mixture into an Oil mixture comprising an oil obtained from sesame heated at a temperature not exceeding 100 C and an oil obtained from raw sesame, wherein a ratio of the oil mixture to a total amount of the powder mixture and the oil mixture is 60 to 95 % by weight.

As the raw material abundantly containing the 6 antioxidant substances, in addition to said rice germ, wheat germ and soybean, there may be listed plant species described in the Japanese Unexamined Parent Application Laid-open No. 63-79834, i.e., barley, adzuki bean, corn, hatomu-i (pearl barley), pea and the like and the like, however, according to the study by the present inventors, the rice germ, wheat germ and soybean are the most preferable raw materials.

Next: to rice germ, wheat germ and soybean, there can be listed rice bran, hatomugi (pearl barley) and wheat. Therefore, as a raw material, at least one of rice bran, hatomugi (pearl barley) and wheat may be included together 'u.-ith rice germ and/or wheat germ. - In any case, the ratio of the germ (rice germ and/or wheat; 0 germ) in the cereal raw material is preferably at least more than or equal to 1 % by weight.

The antioxidant substances in the cereal raw material described above form a complicated macromolecule polymer together with other substances, and therefore has no activity as it is, so that it is necessary to cut macromolecule bonds by means of mildly heating or the like so as to release low molecular weight antioxidant substances. However, if heating temperature is too high, the low molecular weight antioxidant substances are deactivated, so that it is 7 necessary to 'select a heating condition taking into consideration this fact.

In order to satisfy such a condition, it is necessary to heat the cereal raw material at a temperature not exceeding 100 0 C slowly and in sufficient length of time. In detail, the cereal raw material is put in a vessel made of ceramic such as pottery or the like irradiating far-infrared rays, preferably having wavelengths of 4 to 14 -1m, and heating is performed while slowly stirring and.maintaining the temperature at about 90 to 960C.

The heating time varies depending on kinds of cereal species, so that it cannot be indiscriminately defined, however, it is preferably about 30 minutes to 3 hours. Incidentally,Alhe heating method is not limited to the above-mentloned method provided that the antioxidant substances in the cereal raw material are sufficiently released as low molecular substances while deactivation thereof is _prevented.

After heat treatment described above, koji (Aspergillus orizae) is added to the raw material to brew it. The brewing condition is preferably at 20 to 36C for about 2 to 6 days. When brewing is performed using a fermenter, about 2 to 3 hours are sufficient. This brwing is performed in order to further promote 8 liberation and converston into low molecular weight substances of the antioxidant substances in the cereal raw material, and, through this brewing step, the activity of the antioxidant substances are remarkably enhanced as compared with a raw material only subjected to heat treatment.

Next, the brewed material is ground to make it into fine powder. Grinding may be performed using a commercially available grinder. Some types of grinder, however, generate high temperature during the use, thus deactivatinb- the antioxidant substances. Therefore, it is Dreferable to use those which generate no high temperature %hen used. For exanuiple, a stone mill may be used.

Next, the oil is prepared by mixing an oil obtained from heated sesame (hereinafter referred to as the sesame paste oil) with an oil obtained from raw sesame in an appropriate ratio, and the above-mentioned fine powder is added to the oil mixture. The sesame paste oil is an oil obtained by, after a raw sesame is heated at a temperature nor exceeding 100C slowly In sufficient length of time, griding and squeezing it, however, the fine solid contents formed by grinding sesame remain as it is, so that the appearance provides a paste state. This sesame paste oil abundantly contains the low molecular weight antioxidant substances, and by using it, it is possible to obtain the oily preparation having a high activity of the antioxidant substance.

However, this sesame paste oil has a high viscosity. and the size of an oil droplet is alo large, so that the preparation in which only the sesame paste oil is added with the above-described fine powder has a poor penetrating ability into the insides of cells in a disease portion. However, when the sesame oil obtained from raw sese,,-!ie is added to the sesame paste oil, the size of the droplet can be small, and the penetrating ability into the insides of the cells in a disease portion is improved.

The oil collected from raw sesame is an oil obtained by, after a raw sesame is ground as it is and squeezing it, removing solid contents, and a commercially available as an ordinary sesame oil. Since the mixin- ratio of the sesame paste oil to -the ordinary sesame oil is different also depending, on the amount of the fine powder to be added, it cannot be indisciminately defined. It is preferably 1 to 3 parts by weight of the ordinary sesame oil with respect to 1 part by weight of the sesame paste oil.

The ratio of the oil mixture to the fine powder added thereto is such one that the mixed oil conies to 6o to 95 % by weight of a total amount of-the both. If the oil mixture is less than 60 % by weight, the ability to pass through the cell membrane is poor. On the other.hand, if the oil mixture exceeds 95 % by weight, the concentration of the antioxidant substances as the effective components is low, so that the effect of suppressing the production of the reactive oxygen and 11pid peroxide also decreases.

The oily preparation of the present invention, which is orally taken, can be prepared by adding the fine powder to the oil mixture by enclosing it in a gelatin capsule or the like as soon as the oil mixture is prepared. The material mixt-ure, however, can be matured preferably at 20 to 35 0 C for about 3 to 30 days, more preferably at 28 to 30 0C, for about 1 week, before encapsulation.

By applying the maturing treatment. the liberation and low molecular weight conversion of the antioxidant substances are further progressed by the koji remaining in the fine powder, and the antioxidant substances well attaches to the oil mixture, so that the activity and the osmotic ability into the inside of the cell of the reactive components are further improved.

The oily preparation of the present invention can also include a cereal raw material comprising rice germ or wheat germ and soybean, and optionally at least one of rice bran, hatomugi (pearl. barley) or wheat, which Is made into fine powder as it is without heating C and brewing, and added to the oil mixture together with the heated and brewed fine powder described above.

In the fine powder of the cereal raw material which is not heated and brewed, the antioxidant substance as th.e effective components are not liberated and converted into low molecular weight substances. However, when it is added to the oil mixture together with the heated and brewed fine powder, the liberation and low molecular weight conversion of the antioxidant substances gradually proceed even after encapsulation, owing to the koji remaining in the brewed fine powder, so that there is such an advantage that the effect of suppressing the production of the reactive oxygen and lipid peroxide is maintained for a long period.

On the contrary, in the oily preparation in which only the heated and brewed fine powder is used, due to enzyme reactions and the like which continuously proceed even after encapsulation, the antioxidant substances as the effective components are gradually decomposed, so that the effectiv.e period is short as compared with one in which the non-treated fine powder 12 is added.

However, in the case where the adding amount of the non-treated fine powder is too much, the activity of the effective component decreases. Therefore, it is preferable that the nontreated 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, also in this case of including two types of 'Line powders, the oil mixture is 60 to 95 % by weight of the total amount.

In the thus-obtained oily preparation according to the present invention, both the acti,,.-ity oll' the antioxidant substances and the osmotic power into the inside of the cell are hi,-h, and as clarified also from results of clinical tests as descri-bed hereinafter, it exhibits remarkable effects against various inflammations and inveterate diseases for which the therapeutic effects have been insufficient by conventional anti-inflammatory agents.

The oily preparation of the present invention may be encapsulated with gelatin or the like, and orally taken as a medicine. In addition, in the case of abnormal pigmentation such as dermatitis, chloasma, freckles and the like, or wrinkles and the like, the oily preparation can be applied directly to an affected part. In addition to dermatitis and the like, the 13 present oily preparation can be applied also to adult diseases and inveterate diseases such -as chronic arthrorheumatism, thrombophlebitis, progressive systemic sclerema, Buerger's disease, Raynaud's disease, intractable dermatoulcer and the 'Like. The preparation of the invention also has effects on treatment and prevention of other various pollutioncaused diseases, burns, external wounds, fatigue, hangover, constipation and the like.

In addition, the oily preparation of the present invention has no sideeffect because of the use of only cereal, koji and sesame as the raw materials, so that it is possible to take orally as a healt-h food for maintaining and enhancing the health. It is needless to say that, when making a preparation, it is optional to add auxiliary medicines or components useful for health such as various vitamins, mineralLs and the like. and flavors, taste-adjusting agents, coloring agents and the like. Embodiment Each one part of rice germ, soybean, rice bran, hatomugi (pearl barley) and wheat was charged in a pot made of potte ry emitting far-infrared rays of 4 to 14 a m, and heatedAt 90 to 96 C for 3 hours with slow stirring riot to making scorch. Next, kojl was added by 14 3 % of a total amount to brew at 36 to 400C for 72 hours, and then the brewed material was made -into fine powder using a stone mill. In addition, above-mentioned five kinds of cereal raw materials, which were not heated and brewed, were ground with a stone mill to obtain fine powders of the same amount. On the other hand, sesame was heated in the same manner as the above serial materials, and, after it was ground with a stone mill, squeezed to obtain a sesame paste oil.

Next, 28.1 parts. by weight of the sesame paste oil was mixed'with 48.9 parts by weight of a commercially available sesame oil; to which mixture were added 11.5 parts by weight of the heated and brewed fine powder and 11.5 parts by weight of the non-treated fine powder; and the obtained mixture was stirred homogeneously. The obtained mixture was matured at 28 - C for 1 week, then encapsulated in gelatin to obtain an oily preparation. Effect of the Invention [I] In vitro test (1) For an in vitro test of the ability of arriving and penetrating to oily sites, the oily preparation of the embodiment was added to a TBA (thiobarbituric acid) reaction system in which an oily unsaturated fatty acid (decosahexaenolc acid) was reacted with reactive oxygen generated by uitravio.let rays and pr'oduced lipid peroxide, and the degree of suppression of the production of lipid peroxide was measured.

Namely, to 0.1..cc of decosahexaenoic acid diluted by 100 times, 1.8 mg/ml of the test sample of the oily preparation of the present invention was added, and the lipid peroxide produced was measured with the TBA reaction. In the TBA reaction, 0.2. ml of 7 % sodium dodecyl sulfate, 2 ml of 0.1 -X HCl, and 0.3 ml of phospho tung stic. acid were mixed; 1.8 mg/ml of the test sample was added to the mixture; 1 mI of a reagent in which 0.67 % TBA was mixed with acetic acid by 1:1 was added; and the measurement was performed with excitation at 515 nm and emtss.,on at 553 n-m using a fluorescent spectrophoto,meter.

As comparative examples, used the votanical nutrient of Japanese Patent o. 13G32168 (Comparative example 1) and the antioxidant composition decribed in Japanese -Sonexamined Patent Application Laid-open -No. 6379834 (Comparative example 2), measurements being performed in the same manner. Results are shown in Table 1.

1 Table 1

Average Test sample (6 munutes value) Control 461 _+ 6_.} (UV+) oily preparation of the 1-91 13 present invention (1.8 - mg/M1) Comparative example 1 271 34 (1.8 mg/ml) Comparative example 2 358 45 (1.8 mg/M1) UV+: ultraviolet irradiation 0.01 < P < 0.05 (v.s. control) P < 0.01 (v.s. control) P < 0.0001 (v.s. control) - Test result - - Although any of tile test samples significantly 17 suppressed the generation of the lipid peroxide (TBA reactive substances) from the unsaturated fatty acid (docosahexaenoic acid) by irradiation of ultraviolet ray Op-), especially the oily prepara Vion of the present invention remarkably suppressed this generation (P < 0.0001). This result supports the fact that the oily preparation of the present Invention has a higher activity of the antioxidant substance, and a h!,-her penetrating ability into the inside of the cell in which a disease occurs than the conventional preparations (Comparative examples 1 and 2).

(2) A [ M- 3H2]-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 (-3H); it was added into a human tissue In a test tube; and the count (Cpm) of [3 H2] bound to its cell membrane was measured by a scintillation counter; thereby the ability to arrive at the cell membrane was tested (labeled tritiated thymidine was 2 Ci/mM).

The same tests were also performed for Comparative examples 1 and 2. Results are shown in Table 2. As clarified from Table 2, the oily preparation of the present Invention had the highest 18 affinity to cell mem.brane as compared with Comparative examples 1 and 2.

Table 2

Test sample cpm incorporation Control 15643 cpm / 103 cells Oily preparation of 56372 epm / 103 cells the present invention (1.8 mg1M1) Comparative example 1 33451 Cpm / 103 cells (1.8 mg/ml) Comparative example 2 30567 epm. / 103 cells (1.8 mg1M1) (3) The oily preparation of the present Invention was subjected to ultrasonic treatment and added to a reactive oxygen generating system (neutrophile and xanthine-xan thine oxidase) so that a converted living body concentration became 1.6 mg/ml; and three types of 19 reactive oxygens (02-, H202 and OH.) were measured to compare with the case of no addition (control). The converted living body concentration Is an amount assumed to exist in blood when an ordinary taking amount per one day (9 g in the case of the oily preparation of the present invention) is absorbed into a living body.

The measuring methods for the three types of reactive oxygens are as follows.

For 02-, the method was used in which the amount of reduction of f er rieyto chrome C bY 02- was measured at a wavelength of 550 nm with a spectropho tome ter of Beckman, and converted to the amount of Oz-.

For H--02, depending on the fact that H202 decreases fluorescence generated by scopoletin under the presence of peroxidase, by using scopoletin and peroxidase, the degree of decrease in fluorescence of scopoletin was measured using a fluorescent spectrophotometer made by Hitachi Ltd. with excitation of 370 nm and emission of 460 um.

For UP, utilizing a principle that a-ketomethiol-butylic acid (KY1B) reacts with OH, to generate ethylene gas, the method was used in which the ethylene gas was quantifie d with a gas chromatography made by Hitachi Ltd., so as to convert Into 09. The same tests were also performed for Comparative examples 1 and 2.

9 Results are shown in Table.3.

Table 3

Reactive oxygen Test sample 02- H202 OH.

Control 1.532 nmol 485 pmol 854 pmol Oily preparation 0.50 nmol 161 pmol 283 pmol of the present invention (1 - 8 mg1M1) Comparative 1.285 nmol 403 pmol 707 pmol example 1 (1.8 mg/mi) Comparative 0.231 nmol 538 pmol. 108 pmol example 2 (1.8 IngIM1) Test result 21 The anti.oxidant effect of the oily preparation of the present invention was slightly-inferior to that of ComparatIve example 2, however, there was exhibited the antioxidant effect which was stronger than that of Comparative example 1. However, as clarifie'd from abovementioned Tables 1 and 2, Comparative example 2 has a smaller contents of oily subst-ances, so that it has poor ability to arrive at. the cell in which a disease occurs.

Therefore, when it is judo-ed as a total, the oily preparation of the present invention, in which the antioxidant effect is hil-,.h and the penetrating ability into the inside of the cell in a disease portion is highest, is most excellent as an antioxidant agent.

[III Clinical test The therapeutic effect was investigated on 96 patients of autoimmune diseases, collagen diseases such as chronic arthro-rheumatism, hemasthenosis, nephritis, hepatocirrhosis, chloasma, freckles and the like, who have hitherto resisted or got worse. against any one of non- steroidal antiphlogistic drugs, steroids and the antioxidant composition of Comparative example 2. The results are shown in Table 4.

22 Table 4

Test sample Oily preparation Comparative Comparative of the present example 1 example 2 invention (1.8 mg/ml) (1.8 mg/mi) Disaese (1.8 mg1M1) Chronic 13/18 (72%) 0/7 (0%) 4/14 (28%) arthro rheumatism AngHtis 3/5 (GOM 0/3 (0%) 1/4 (25%) Progressive 9/12 (75%) 2/10 (20%) 6/12 (50%)' systemic selerema Dermato- 6/8 (715%) 2/12 (16 %) 6/13 (4G%) myosytis Thrombo- 2113 (G 6 0/4 (006) 1/5 (201OU1) phlebitis Buerger's 6/8 (75%) 0/3 (OM 3/6 (50%) disease Raynaud's 10/12 (83%) 3/8 (37%) 7/12 (58%) disease Crus 8/11 (72%) 4/9 (44%) 11/15 varicosIs Intractable 3/5 (60%) 1/5 (20%) 2/6 (3 31k) dermatoulcer Chloasma. 11/14 (78%) 4/12 (33%) 17/25 (68%) freckles 23

Claims

CLAIMS 1 An oily preparation obtainable by:

1 heating a cereal raw material comprising rice germ and/or wheat germ and soybean at a temperature not exceeding 100OC; making the heated material Into fine powder; adding koji to the powdered material and brewing the powdered material; and adding thus-brewed material to an oil mixture comprising a paste oil obtained.Prom sesame heated at a temperature nou exceeding 100 0 C and an oil obtained from raw sesame, wherein a ratio of said oil mixture to a total amoun.t of the fine powder and the oil mixture is to 95 10,0 by weight.

2.1 The oily prepc-ration according to claim 1. which further includes a cereal raw material comprising rice germ and/or wheat germ and soybean made Into fine powder without heating and brewing.

3. The oily preparation according to claim 1 or 2 wherein said heated or non-heated cereal raw mater-lal Includes, in addition to rice germ and/or wheat germ and Soybean, at least one of rice bran, hatomug! (pearl 24 barley) or wheat.

4. The oily preparation according to claim 1 or 2 which is encapsulated for taking orally as a medicine or a health food.

A method of producing an oily preparation wherein when the cereal raw material of claim 1, 2 or 3 is heated with far-infrared ray having a wavelength of 4 to 14 MI m.

6. A method of producing an oily preparation wheretn the cereal raw material according to claim 1, 2 or 3 is brewed at 20 to 360C for 2 to 6 days after treatment with heat.

7. A method of producing an oily preparation wherein the fine powder according to claim 1, 2 or 3 is added to the oil mixture, so as to mature at 9-0 to 35t for 3 to 30 days.

8. An oily preparation according to claim 1, substantially as hereinbefore described.

9. A method according to claim 5, substantially as hereinbefore described.