WO2003070022A1 - Free radical quenching agent contaiing eriobotryae seed extract as the active ingredient and health foods and drinks containing eryobotryae seed extract - Google Patents

Free radical quenching agent contaiing eriobotryae seed extract as the active ingredient and health foods and drinks containing eryobotryae seed extract Download PDF

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Publication number
WO2003070022A1
WO2003070022A1 PCT/JP2003/001951 JP0301951W WO03070022A1 WO 2003070022 A1 WO2003070022 A1 WO 2003070022A1 JP 0301951 W JP0301951 W JP 0301951W WO 03070022 A1 WO03070022 A1 WO 03070022A1
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Prior art keywords
loquat
extract
drink
health food
sitosterol
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PCT/JP2003/001951
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French (fr)
Japanese (ja)
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Yutaka Nishioka
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Techno Network Shikoku Co., Ltd.
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Priority claimed from JP2002046812A external-priority patent/JP2003245056A/en
Priority claimed from JP2002047539A external-priority patent/JP2003246745A/en
Application filed by Techno Network Shikoku Co., Ltd. filed Critical Techno Network Shikoku Co., Ltd.
Priority to AU2003211250A priority Critical patent/AU2003211250A1/en
Publication of WO2003070022A1 publication Critical patent/WO2003070022A1/en

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/52—Adding ingredients
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives

Definitions

  • the present invention relates to a free radical scavenger, and more particularly to a free radical scavenger containing loquat core extract. Further, the present invention relates to a health food and a health drink, particularly to a health food and a health drink containing loquat nucleus extract.
  • extracts having specific components extracted from natural substances such as animals and plants have various effective effects.
  • fish oil extracts and soybean extracts are known to be effective for hyperlipidemia.
  • Natural substances generally have good intestinal absorption, have no side effects, and are highly safe for the human body.
  • loquat is commonly planted widely as a fruit tree, fruits are used for food, and leaves are used for skin diseases, but seeds (loquat nuclei) are used as waste. It has been processed and is not being used effectively. It is not known that loquat seeds (loquat nuclei), a kind of Rosaceae plant widely planted as ordinary fruit trees in Japan, have a more effective action than Kampo extract. Therefore, it would be meaningful if active ingredients could be extracted from loquat nuclei by any method and a safe health food or beverage could be provided without side effects by using each of the active ingredients. However, almost no such effective extract from loquat nucleus is known. Then, an object of the present invention is to provide a useful scavenger capable of scavenging free radicals without side effects. Another object of the present invention is to provide a health food and a health drink containing many active ingredients.
  • the inventors focused on extracts derived from seeds of various plants, and particularly studied the free radical scavenging effect of loquat nucleus extract extracted from loquat nucleus. As a result, they have found the scavenger of the present invention. In addition, as a result of examining the active ingredient of loquat nucleus extract extracted from loquat nucleus, a health food and a saliva drink of the present invention were found.
  • the free radical scavenger of the present invention comprises a loquat nucleus-derived extract as an active ingredient.
  • the loquat nucleus-derived extract comprises a ground material obtained by pulverizing loquat nucleus, comprising ethanol, methanol, water, hexane, or a mixture thereof. It is obtained by immersing in at least one solvent selected from the group and collecting the supernatant.
  • the loquat nucleus-derived extract is selected from the group consisting of: linoleic acid, linolenic acid,
  • the loquat nucleus-derived extract comprises: 3) -sitosterol and i3-sitosterol-3-0-monoglycoside.
  • the health food or drink of the present invention is characterized by containing a loquat core-derived extract.
  • the loquat nucleus-derived extract is obtained by milling a loquat nucleus obtained by powdering the loquat nucleus from ethanol, methanol, water, hexane, or a mixture thereof. And immersing it in at least one solvent selected from the group consisting of:
  • the loquat nucleus-derived extract comprises linoleic acid, linolenic acid, / 3-sitosterol, / 3-sitosterol-3-0-monoglycoside, amygdalin, benzaldehyde , Mandenitrile, and benzoic acid.
  • the loquat nucleus-derived extract contains ⁇ -cytostelol and 0-sitosterol 3-O_monoglycoside.
  • the loquat nucleus-derived extract further contains linoleic acid.
  • the loquat nucleus-derived extract further contains linolenic acid.
  • the health food or drink is a health food.
  • preferred embodiments of the health food and drink of the present invention further include rice flour, buckwheat flour, It contains at least one member selected from the group consisting of udon flour, flour, and bread crumbs.
  • the health food or drink of the present invention is a health drink.
  • a preferred embodiment of the health food or drink of the present invention is characterized in that the health food or drink further contains water, cocoa, coffee, tea, milk, sake, fruit drink, or vegetable drink.
  • FIG. 1 is a diagram showing an example of a method for preparing loquat nucleus extract.
  • FIG. 2 is a diagram showing the results of thin-layer chromatograms when various solvents were used.
  • Figure 3 A when using the active oxygen production stimulator f ML P, activity oxygen from neutrophils (0 2 -) shows the effect of loquat nuclear extract and its merit components on production.
  • FIG. 3B shows the effect of loquat nucleus extract and its active components on the production of active oxygen (O 2 —) from neutrophils when the active oxygen production stimulant f MLP is used.
  • Figure 3 C when using the active oxygen production stimulator f ML P, shows the effect of loquat nuclear extract and its merit components on the activity oxygen (0 2 _) production from neutrophils.
  • FIG. 3H shows the effect of loquat nucleus extract and its active components on the production of active oxygen (O 2 —) from neutrophils when the active oxygen production stimulant f MLP was used.
  • Figure 3 I when using the active oxygen production stimulator f ML P, shows the effect of loquat nuclear extract and its merit components on the activity oxygen (0 2 _) production from neutrophils.
  • FIG. 4A shows the effect of radicat and ascorbic acid on the production of active oxygen (O 2 ⁇ ) from neutrophils when the active oxygen production stimulant f MLP was used.
  • Fig.5A shows that active oxygen from neutrophils
  • Fig. 5B shows that active oxygen from neutrophils
  • loquat nucleus extract and its active ingredient on ( ⁇ 2 —) production The effect of loquat nucleus extract and its active ingredient on ( ⁇ 2 —) production is shown.
  • Fig. 5C shows active oxygen from neutrophils when the active oxygen production stimulant PMA was used.
  • Fig. 5D shows that reactive oxygen from neutrophils
  • Fig. 5E shows active oxygen from neutrophils when the active oxygen production stimulating agent PMA was used.
  • Fig.5F shows active oxygen from neutrophils when active oxygen production stimulating agent PMA was used.
  • Fig.5G shows that active oxygen from neutrophils
  • Fig. 5H shows the active oxygen from neutrophils when the active oxygen production stimulating agent PMA was used.
  • Fig. 5I shows that active oxygen from neutrophils
  • Fig. 6A shows active oxygen from neutrophils when the active oxygen production stimulant PMA was used.
  • FIG. 6B shows that active oxygen from neutrophils
  • Fig. 7A shows active oxygen from neutrophils when active oxygen production stimulant A A was used.
  • loquat nucleus extract and its active ingredient on (0 2 ") production The effect of loquat nucleus extract and its active ingredient on (0 2 ") production is shown.
  • Fig. 7B shows that active oxygen from neutrophils
  • Fig.7C shows that active oxygen from neutrophils
  • Fig. 7D shows active oxygen from neutrophils using the active oxygen production stimulant PMA.
  • Fig.7F shows that active oxygen from neutrophils
  • Fig.7G shows that active oxygen from neutrophils
  • Fig. 7H shows that active oxygen from neutrophils
  • FIG. 71 shows the effect of radicat and ascorbic acid on the production of active oxygen ( ⁇ 2 ) from neutrophils when the active oxygen production stimulant PMA was used.
  • Fig. 8A shows that active oxygen from neutrophils
  • Fig. 8B shows that active oxygen from neutrophils
  • Figure 9 shows the biological test values (GOT, GPT, total cholesterol, LDL cholesterol, HDL cholesterol, and tridalicelide) of normal rabbits.
  • FIG. 9 shows the biological test values (GOT, GPT, total cholesterol, LDL cholesterol, HDL cholesterol, and tridalicelide) of normal rabbits.
  • FIG. 10 is a diagram showing the results of examining the biochemical test values of rabbits to which a feed containing 1% cholesterol monocle was administered.
  • FIG. 11 is a diagram showing the results of examining biochemical test values of hyperlipidemic rabbits.
  • FIG. 12 is a diagram showing the results of examining biochemical laboratory values of hyperlipidemic rabbits.
  • Figure 13 shows the biological test values (GOT, GPT, total cholesterol,
  • FIG. 14 is a diagram showing the results of examination of biochemical test values of rabbits to which a diet containing 1% cholesterol was administered.
  • FIG. 15 shows the results of examining the biochemical test values of hyperlipidemic rabbits.
  • FIG. 16 is a diagram showing the results of examining the biochemical test values of hyperlipidemic rabbits.
  • the free radical scavenger of the present invention contains loquat core extract.
  • Loquat core extract is an extract derived from loquat seeds.
  • the loquat nucleus extract applied to the present invention covers all loquat nucleus extracts as long as they are derived from loquat seeds.
  • a free radical is also called a free radical and means an atomic group that has a wide unpaired electron.
  • molecules with free radicals and substances that produce free radicals are highly reactive and harmful to cells. Given DNA, it damages DNA and induces mutations. Such free radicals can be efficiently removed in the present invention.
  • FIG. 1 is a diagram showing an example of a method for preparing loquat nucleus extract. Loquat nuclei are washed and dried if necessary. Drying is preferably performed sufficiently. The subsequent grinding is carried out homogeneously. To do so.
  • the method of pulverization is not particularly limited, and a known pulverizer such as a pole mill, a hammer mill, a roller mill, a rod mill, a sample mill, a stamp mill, a date integrator, a mortar, and a blender with a cooling device can be used. It is to be noted that since the loquat nucleus composition may be decomposed due to heat generated during the pulverization, a blender with a cooling device is preferable.
  • the solvent in this case is not particularly limited, and a solvent can be appropriately set according to a desired effect.
  • the solvent include any polar or non-polar solvent such as ethanol, methanol, water, hexane, ethyl acetate, chloroform, and acetone. From the viewpoint that loquat nucleus extract having high cell membrane permeability can be obtained, methanol, ethanol, water and the like are preferable.
  • the immersion can be performed under gentle stirring.
  • the above-mentioned crushed material is immersed in various solvents to obtain various solutions.
  • Various solutions may be stirred according to the state of the solution, and the solution may be left as it is in some cases.
  • stirring there is no particular limitation.
  • loquat nucleus extract powder Thereafter, the supernatant is collected and evaporated to dryness to obtain loquat nucleus extract powder. Evaporation to dryness can be performed in a warm bath at 55 ° C to 80 ° C using an evaporator.
  • loquat nuclei Components contained in loquat nuclei are sorted by their polarities by extracting loquat nuclei with solvents of different polarities. Therefore, the type and content of the components of the loquat core extract differ depending on the solvent used.
  • FIG. 2 shows thin layer chromatograms (TLC) of loquat nucleus extract extracted using various solvents to examine the components of various loquat nucleus extracts.
  • the extract in which the solvent is water (hereinafter referred to as the water extract) Say. ) Shows a spot only at the origin, and is considered to contain highly polar compounds such as tampa, sugars and amigdarin.
  • Extracts in which the solvent is 70% ethanol (hereinafter referred to as 70% ethanol extract) and extracts in which the solvent is methanol (hereinafter referred to as methanol extract) have water spots at the origin in the thin-layer chromatogram. It is thought that there are few highly polar compounds such as proteins, sugars, and amygdalin, which are smaller than extracts.
  • Extracts with hexane as the solvent show compounds with an R f value of 0.71 ⁇ in thin-layer chromatograms and contain many low-polarity compounds.
  • the components were analyzed using a magnetic resonance method (NMR), a mass spectrometry method (MS), an ultraviolet absorption spectrum method (IR), and the like.
  • NMR magnetic resonance method
  • MS mass spectrometry method
  • IR ultraviolet absorption spectrum method
  • at least eight active ingredients, linoleic acid, linolenic acid,] 3-sitosterol, j6-sitosterol-3-0-monoglycoside, amygdalin, benzaldehyde, mandenitrile, and benzoic acid, and a proline-like compound were confirmed. .
  • the compounds isolated from Loquat Nucleus Extract are shown in Table 1 below.
  • the pharmaceutical composition containing the loquat nucleus extract of the present invention contains at least one of the above eight active ingredients.
  • the pharmaceutical composition containing loquat nucleus extract of the present invention preferably contains] 8-sitosterol and j8-sitosterol-13-0-monoglycoside.
  • 3-sitosterol-1-3-0-monoglycoside have a free radical scavenging action and a hyperlipidemia-improving action directly.
  • the pharmaceutical composition containing the loquat nucleus extract of the present invention may contain, in addition to] 3 sitosterone and 3) -sitosterol-3-0-monodalicoside, linoleic acid, and Z or linolenic acid. preferable. Linoleic acid and linolenic acid have a cholesterol dissolving action.
  • the free radical scavenger according to the invention is prepared in an effective amount of loquat nucleus extract and in a suitable dosage form.
  • the dose of loquat nucleus extract in the free radical scavenger of the present invention can be changed depending on the condition and severity of the subject to be administered, the dosage form, the selected administration route, the number of administrations per day, and the like.
  • the dose of loquat nucleus extract in the free radical scavenger of the present invention should be at least 375 mg / kg / day in rats, and even lower in humans due to differences in sensitivity. Is preferred.
  • oral preparations tablettes, capsules, coated tablets, granules, solutions, syrups
  • suppositories for rectal administration, injections and the like
  • patients to be administered are often oral drugs from the viewpoint of the necessity of long-term administration and the easiness of operation, which are often chronic diseases.
  • Administration can include other conventional ingredients such as stable preservatives, sweeteners, coloring agents and flavoring agents.
  • the lethal dose of loquat nucleus extract can be estimated to be 8060 mgZkg based on its amygdalin content (7.4%).
  • loquat core used was loquat seeds collected in Kochi Prefecture and dried sufficiently by sun drying.
  • Loquat nucleus extract is obtained by immersing loquat nucleus (1,050 g) ground (1,000 rpm) in a blender with a cooling device in twice the volume of 70% ethanol (2,1000 m1), and constantly using a stirrer. After stirring (300 rpm), the supernatant was collected on the 7th day after the immersion, and evaporated to dryness in a hot bath at 70 ° C in an evaporator to produce the product.
  • loquat nucleus extract was prepared (Fig. 1), and the components were fractionated using solvent separation method, silica gel column chromatography, molecular sieve column chromatography, and thin layer chromatography (TLC), and eight kinds of components were separated. The components were isolated. Comparison of these compounds with samples using high performance liquid chromatography (HPLC), TLC, etc., and nuclear magnetic resonance (NMR), mass spectrometry (MS), infrared absorption spectrum (IR) ) was determined by instrumental analysis.
  • HPLC high performance liquid chromatography
  • TLC molecular sieve column chromatography
  • TLC thin layer chromatography
  • IR infrared absorption spectrum
  • the loquat core extract contained / 3-sitosterol,] 3-sitosterol 3-0-monoglycoside, linoleic acid, linolenic acid, amygdalin, benzaldehyde, mandenitrile and benzoic acid.
  • the results of the identification are shown below. The measurement conditions are as follows.
  • 1 / 3-0-Glycoside was isolated from loquat nucleus extract and identified by comparison with the standard data in 1 H NMR (bi, CDC13) and 13 C-NMR (bi, pyridine-d5) spectra. .
  • Benzoic acid was isolated from loquat nucleus extract and identified by comparison with standard data in 1R, HR-EI-MS, 1HNMR (CDC13) and HR-EI-MS spectra.
  • loquat nucleus extract was subjected to HP LC, and compared with the authentic sample. In the HPLC of loquat nucleus extract, a peak showing the same retention time as the linoleic acid and linolenic acid samples was observed. HPLC measurement conditions for linoleic acid and linolenic acid Hitachi D600 High Performance Liquid Chromatograph
  • loquat nucleus extract was subjected to HP LC and identified by comparison with the standard.
  • the health food or health drink of the present invention contains loquat core extract.
  • Loquat core extract is an extract derived from loquat seeds.
  • the loquat nucleus extract applied to the present invention covers all loquat nucleus extracts as long as they are derived from loquat seeds.
  • healthy food or health drink is not particularly limited, and broadly means a food or beverage containing loquat nucleus extract.
  • the loquat nucleus extract of the present invention can be used as it is. However, it can be mixed with other natural ingredients as described below for edible use.
  • the method for adjusting loquat nuclei is as follows. About 1 to 10 mg of the powder derived from loquat nucleus extract obtained as described above can be dissolved in 10 to 100 ml of water and eaten. You can take this 2-3 times a day.
  • the health food containing loquat extract can be used as a meal substitute or snack.
  • the powder derived from the loquat core extract is rice flour, buckwheat flour, udon flour, Wheat flour or at least one selected from bread crumbs can be mixed and used for ordinary cooking. These may be processed into Western confectionery, Japanese confectionery, confectionery, udon, buckwheat noodles, pasta, Chinese noodles, bread, etc. to make them even easier to eat. These cooking methods follow common law.
  • the health food may be in a solid (powder, granule), jelly, liquid, or suspension form.
  • the health food or the health drink may contain various components contained in a sweetener, an acidulant, a vitamin, a drink and the like.
  • it may further contain at least one selected from the group consisting of water, cocoa, coffee, tea, milk, alcohol, fruit drinks, and vegetable drinks.
  • the method of eating and drinking the health food or the healthy beverage of the present invention is free and not particularly limited. Therefore, the effects of the present invention can be expected as long as the eating and drinking method utilizes food materials. Therefore, the embodiments of the invention and the eating and drinking methods described in the following examples are merely examples of the present invention used for explanation, and should not be construed as limiting.
  • Neutrophils were separated from rat peripheral blood by concentration gradient centrifugation.
  • the isolated neutrophils are treated with the active oxygen production stimulant FML P (N-formy methiony
  • the active oxygen scavenging ability was represented by an active oxygen production suppression rate and an active oxygen removal rate determined by a ratio between a group administered with loquat nucleus extract and its active ingredient and a group not administered.
  • Figures 4 (a) and 4 (b) show the control of radicat and ascorbic acid, respectively.
  • the loquat nucleus extract and; 8-sitosterol of the present invention showed excellent active oxygen production suppressing effect and active oxygen removing effect.
  • Radicut showed a weak active oxygen production inhibitory effect, but did not show an active oxygen removing effect.
  • Ascorbic acid did not show an active oxygen production inhibitory effect, but showed a weak active oxygen removing effect.
  • Example 2 a test was conducted in the same manner as in Example 1, except that PMA (Phorbol myristate acetate) was used as a reactive oxygen production stimulating agent instead of fMLP.
  • PMA Phorbol myristate acetate
  • Figures 6 (a) and 6 (b) show the results of Radicut and ascorbic acid in the control, respectively.
  • the loquat nucleus extract of the present invention exhibited an active oxygen production suppressing effect and an active oxygen removing effect.
  • ⁇ -sitosterol showed a weak active oxygen removal effect, and no active oxygen production inhibitory effect.
  • Radicut showed very weak active oxygen production inhibitory effect and active oxygen removal effect, but ascorbic acid had no effect.
  • the loquat nucleus extract of the present invention exhibited an active oxygen production inhibitory effect when the concentration was increased. In addition, it showed an active oxygen removal effect.
  • 3-Sitosterol showed weak active oxygen production inhibitory effect and active oxygen removal effect.
  • Radicut on the other hand, showed a slight inhibitory effect on active oxygen production and no active oxygen removal effect. Ascorbic acid had no effect.
  • the active oxygen (0 2 —) scavenging ability of loquat nucleus extract was examined by using a highly water-soluble tetralithodium salt reduction method.
  • the loquat nucleus extract of the present invention had a high SOD activity of 85% at a concentration of 10 OM, but the active ingredients, Radicut and ascorbic acid were around 10% or were not observed.
  • Table 3 shows the loquat core extract, its active ingredient, Radicut, and the amount of DPPH 50% eliminated for ascorbic acid.
  • the loquat nucleus extract of the present invention has a DPPH 50% elimination amount of 1%.
  • Radicut and ascorbic acid as controls were also found as low as 13.7 ⁇ and 28.2 ⁇ .
  • ESR Electro Spin Resonance
  • Loquat nucleus extract 3 ⁇ 43 ⁇ 4i ⁇ T. Mouth ”4 ⁇ ⁇ Loquat nucleus extract, its active ingredient, hydroxy radical scavenging activity of dicat and ascorbic acid Loquat nucleus extract and active ingredient scavenging activity (%) Loquat nucleus extract 32.1 ⁇ 3.3
  • Linolenic acid 1 2. 6 ⁇ 1.
  • the hydroxyl radical scavenging activity (ESR method) by the Fenton reaction was as high as 32% for loquat nucleus extract, 27% for ⁇ -sitosterol, and 24% for benzoic acid.
  • the acid showed almost the same erasing activity as 23% and 27%.
  • Hakone extract (10.4 g) was prepared by dissolving it in 400,000 ml of water, and this was eaten twice a day, 100 ml at a time, for one week for one week.
  • Table 5 shows the results. Table 5 shows the effects on subjects whose liver function test values (AST and ALT) are outside the normal range.
  • Loquat nucleus extract powder (10.4 g) was prepared by dissolving it in 400,000 ml of water, and this was eaten twice a day, 10 Oml once a day for 3 weeks. Table 6 shows the results. Table 6 shows the effect on subjects whose liver function test value ( ⁇ -GTP) exceeds the normal range.
  • Loquat nucleus extract powder 10.44 g was prepared by dissolving in 400,00 Om 1 of water, and this was eaten twice daily and 10 Omi once a week for 1 week.
  • Table 7 shows the results. Table 7 shows the effect on subjects whose liver function test values are within the normal range. Table 7
  • the extract content of each aqueous solution was 0.5% for 70% ethanol extract, 0.45% for methanol extract, 0.75% for water extract, and 0.2% for hexane extract, respectively.
  • Rats were sacrificed 7 days after administration of normal rats and hepatic impaired rats after administration of various extracts. Serum glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate Transaminase (GPT) values were measured.
  • GTT Serum glutamate oxaloacetate transaminase
  • GPT glutamate pyruvate Transaminase
  • Fig. 9 shows the results.
  • the G ⁇ T and GPT values in the group receiving the normal data were significantly higher than those in normal rats.
  • the GOT value of the loquat nucleus extract-administered group showed a difference depending on the solvent.
  • it was observed that the 70% ethanol and methanol extract-administered group showed lower values than the normal feed-administered group.
  • the effect was also observed in the group to which water and hexane extract were administered compared to the group to which normal feed was administered.
  • hepatopathy rats and administration of various extracts were performed according to Example 10. Seven days after administration, the amount of retinol palmitate in the liver was measured for normal rats and hepatic impaired rats after administration of various extracts. Approximately 0. 3 g is weighed, 5 ml of black-mouthed form is added, and the mixture is homogenized (10,000 rpm, 2 minutes) using a cell disrupter (manufactured by Eilard) under ice-cooling, and centrifuged (3,500 rpm, 20 rpm). After that, the lower layer was separated and used as a sample. These samples were filtered through a 0.5 ⁇ membrane filter (manufactured by Millipore) to obtain samples for HP LC.
  • a cell disrupter manufactured by Eilard
  • the measurement conditions are as follows. Hitachi L 6000 High Performance Liquid Chromatograph
  • Flow rate 1.0 ml / in Figure 10 shows the results.
  • Liver retinoid content was expressed as a percentage of the liver retinoid content of normal rats immediately after pre-breeding, assuming 100%. Liver retinoid levels were significantly lower in the normal diet group than in normal rats.
  • a difference was observed in the amount of hepatic retinoids in the group administered with the solvent depending on the solvent. Particularly, in the group to which 70% ethanol, methanol and water extract were administered, the amount of hepatic retinoid was increased as compared to the group to which normal feed was administered. No difference was observed in the hexane extract administration group from the normal feed administration group.
  • Preparation of hepatopathy rats and administration of various extracts were performed according to Example 10. Approximately 0.3 g of a fixed part of the rat isolated liver was weighed, 5 ml of ethanol was added, and the mixture was homogenized (10,000 rpm, 2 minutes) using a cell crusher (manufactured by Eilard) under ice-cooling. After centrifugation (3,500 rpm, 20 minutes), the supernatant was collected. 1 ml of this solution was taken out and heated to 60 ° C. for 8 hours to dryness. The residue was dissolved by adding 80 l of ethanol 40 ⁇ 0.
  • the measurement conditions are as follows.
  • FIG. 11 shows the results of 1.0 m 1 / min.
  • the amount of hydroxyproline was significantly higher in the normal diet group than in normal rats.
  • the extract-administered group a difference was observed in the amount of hepatic retinoid depending on the solvent.
  • the amount of hydroxyproline was reduced compared to the group to which normal feed was administered.
  • the hexane extract-administered group a slight difference was observed compared to the normal feed-administered group.
  • liver fibrosis in rats treated with various extracts was calculated by calculating the area of liver fibrosis sites stained blue by computer image analysis from liver tissue images stained with Azan-Mallory staining of isolated rat liver. It was determined from the ratio of the liver fibrosis area to that of a normal feed-treated rat.
  • the results are shown in FIG.
  • the hepatic fibrosis rate was expressed as a ratio to the fibrosis area of the normal diet administration group, which was defined as 100%.
  • the liver fibrosis rate was lower in the group receiving loquat extract than in the group receiving normal feed. Further, the value was different depending on the solvent, and the value was lower in the 70% ethanol and methanol extract administration groups than in the water and hexane extract administration groups.
  • Loquat nucleus extract was prepared according to the above procedure. Cholesterol-containing feed-administered rabbits were preliminarily reared into 6 rabbits per group, and 1% cholesterol-containing feed (manufactured by Shimizu Experimental Materials Co., Ltd.) The loquat nucleus extract was divided into two groups and administered for 4 weeks. The dose of the diet containing 1% cholesterol was 75 g / day (37.5 to 41.7 mg / kg as cholesterol).
  • Loquat nucleus extract was prepared according to the above procedure. First, pre-bred rabbits were treated with 1% cholesterol-containing feed and drinking water for 3 weeks to prepare hyperlipidemic rabbits. The dose of the 1% cholesterol-containing feed was 75 g per day (37.5 to 41.7 mg Z kg as cholesterol). For these rabbits, the feed was changed from 1% cholesterol-containing feed to normal feed, and then 6 rabbits per group were used. They were divided into groups for feed-feeding and drinking water administration, normal feed and loquat nucleus extract administration, and were administered for 4 weeks.
  • the loquat nucleus extract was administered in the same manner as in Example 16 except that hyperlipidemic rabbits were prepared by administering 1% cholesterol-containing feed and drinking water for 4 weeks.
  • Biochemical test values were measured in the same manner as in Example 14. The results are shown in FIG. As is clear from the results in Fig. 16, the serum that is a factor that promotes atherosclerosis when loquat nucleus extract is administered to hyperlipidemic rabbits is greater than when loquat nucleus extract is not administered. Total and LDL cholesterol levels were low.
  • the free radical scavenger of the present invention has an advantageous effect that harmful free radicals can be efficiently removed.
  • the free radical scavenger of the present invention is a natural material, it can achieve desired effects without causing side effects, and can not only contribute to medical treatment but also reduce the range of use of agricultural products. It has the beneficial effects of high social contribution, such as expansion and effective use of waste.
  • the health food or health drink of the present invention has an advantageous effect of being able to maintain and promote health, which cannot be obtained when natural materials are used alone.
  • As an effect to maintain and promote health specifically, it is possible to obtain effects such as prevention and improvement of liver dysfunction, cholesterol dissolving, hyperlipidemia improving, antitussive and expectorant effects without side effects. It has the advantageous effect of turning off.
  • the health food or health drink of the present invention can prevent diseases such as hepatitis, cirrhosis and hypolipidemia in a daily diet by being added to foods or drinks to be taken daily. There is an advantageous effect that improvement can be expected.

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  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

It is intended to provide an advantageous agent whereby free radicals can be quenched without any side effects and health foods and health drinks containing many active ingredients. The free radical quenching agent described above is characterized by containing an eriobotryae (Japan medlar) seed extract as the active ingredient. The health foods and drinks described above are characterized by containing an eriobotryae seed-origin extract as the active ingredient.

Description

明 細 書 枇杷核エキスを有効成分とするフリーラジカル消去剤及び 枇杷核エキスを含有する健康飲食品 技術分野  Description Free radical scavenger containing loquat nucleus extract as active ingredient and health food and beverage containing loquat nucleus extract
本発明は、 フリーラジカル消去剤に関し、 特に、 枇杷核エキスを含有するフリ 一ラジカル消去剤に関する。 また、 本発明は、 健康食品及び健康飲料に関し、 特 に、 枇杷核エキスを含有する健康食品及び健康飲料に関する。 背景技術  The present invention relates to a free radical scavenger, and more particularly to a free radical scavenger containing loquat core extract. Further, the present invention relates to a health food and a health drink, particularly to a health food and a health drink containing loquat nucleus extract. Background art
動物、 植物等の天然物質から抽出した特定の成分を有するエキスが、 様々な有 効作用を有することが知られている。 特に、 動物、 植物等の天然物質から抽出し たエキスの中では、 魚油抽出物及び大豆抽出物は、 高脂血症に有効であることが 知られている。 天然物質は、 一般に腸管吸収に優れ、 副作用がなく、 人体に対す る安全性が高いという特徴がある。  It is known that extracts having specific components extracted from natural substances such as animals and plants have various effective effects. In particular, among extracts extracted from natural substances such as animals and plants, fish oil extracts and soybean extracts are known to be effective for hyperlipidemia. Natural substances generally have good intestinal absorption, have no side effects, and are highly safe for the human body.
一方、 最近では、 好中球等の食細胞が産生する活性酸素により生成される過酸 化脂質が肝線維化の進展に大きく影響を与えることが注目されている。 そのため 肝線維化の抑制には活性酸素 （02— ) 及びハイドロキシラジカル （ · 0Η) 等の フリーラジカルの消去能を有する薬物の投与が有用であり、 種々の薬物について 検討が行なわれている。 On the other hand, recently, attention has been paid to the fact that peroxidized lipids generated by active oxygen produced by phagocytes such as neutrophils greatly affect the progression of liver fibrosis. Therefore the inhibition of hepatic fibrosis active oxygen (0 2 -) and hydroxy radicals (· 0Ita), etc. are useful administration of a drug with a scavenging ability of free radicals, examined various drugs have been made.
また、 高コレステロ一ル血症に伴う動脈硬化や虚血性心疾患の発症にフリーラ ジカルが大きく関与していることが報告されている。  It has also been reported that free radicals are significantly involved in the development of arteriosclerosis and ischemic heart disease associated with hypercholesterolemia.
背景技術 Background art
しかしながら、 天然物質を抽出する場合に、 有効成分を上手に抽出することが 困難な場合もあり、 簡便に有効成分を抽出でき、 かつ様々な有効な作用を有する 天然物質の利用が望まれていた。 However, when extracting natural substances, there are cases where it is difficult to extract the active ingredient well, and the active ingredient can be easily extracted and has various effective effects. The use of natural substances has been desired.
また、 上述したような、 肝線維化、 動脈硬化、 虚血性心疾患等に有害なフリ一 ラジカルを消去し得る消去剤に関しても、 未だ満足な結果が得られていない現状 にある。  In addition, satisfactory results have not yet been obtained for scavengers capable of scavenging free radicals harmful to liver fibrosis, arteriosclerosis, ischemic heart disease and the like as described above.
また、 上述のような天然物質のうち、 枇杷は、 通常果樹として広く植栽され、 果実は食品、 葉は、 皮膚疾患等に用いられているものの、 種子 （枇杷核） は、 廃 棄物として処理されており、 有効に利用されていない。 そして、 本邦において通 常果樹として広く植栽されているバラ科植物の一種である枇杷の種子 （枇杷核） に、 漢方エキスに優る有効な作用があることは知られていない。 したがって、 枇 杷核から、 何らかの方法により有効成分を抽出して、 各有効成分を利用すること により副作用なしに安全な健康食品または健康飲料を提供できれば有意義である。 しかし、 このような枇杷核由来の有効なエキスは、 ほとんど知られていない。 そこで、 本発明は、 フリーラジカルを副作用なしに消去し得る有益な消去剤を 提供することを目的とする。 また、、本発明は、多くの有効成分を含有する健康食 品及び健康飲料を提供することを目的とする。  Of the natural substances mentioned above, loquat is commonly planted widely as a fruit tree, fruits are used for food, and leaves are used for skin diseases, but seeds (loquat nuclei) are used as waste. It has been processed and is not being used effectively. It is not known that loquat seeds (loquat nuclei), a kind of Rosaceae plant widely planted as ordinary fruit trees in Japan, have a more effective action than Kampo extract. Therefore, it would be meaningful if active ingredients could be extracted from loquat nuclei by any method and a safe health food or beverage could be provided without side effects by using each of the active ingredients. However, almost no such effective extract from loquat nucleus is known. Then, an object of the present invention is to provide a useful scavenger capable of scavenging free radicals without side effects. Another object of the present invention is to provide a health food and a health drink containing many active ingredients.
発明の開示 Disclosure of the invention
発明者らは、 各種植物の種子由来エキスに着目し、 特に、 枇杷核から抽出した 枇杷核エキスのフリ一ラジカル消去作用について検討した結果、 本発明の消去剤 を見出すに至った。 また、 枇杷核から抽出した枇杷核エキスの有効成分について 検討した結果、 本発明の健康食品及び俸康飲料を見出すに至った。  The inventors focused on extracts derived from seeds of various plants, and particularly studied the free radical scavenging effect of loquat nucleus extract extracted from loquat nucleus. As a result, they have found the scavenger of the present invention. In addition, as a result of examining the active ingredient of loquat nucleus extract extracted from loquat nucleus, a health food and a saliva drink of the present invention were found.
本発明のフリーラジカル消去剤は、 枇杷核由来エキスを有効成分とすることを 特徴とする。  The free radical scavenger of the present invention comprises a loquat nucleus-derived extract as an active ingredient.
また、 本発明のフリーラジカル消去剤の好ましい態様としては、 枇杷核由来ェ キスが、 枇杷核を粉砕して得た粉碎物を、 エタノール、 メタノール、 水、 へキサ ン、 又はこれらの混合物からなる群から選択される少なくとも 1種の溶媒に浸漬 して、 上清を分取して得られることを特徴とする。 また、 本発明のフリーラジカル消去剤の好ましい態様としては、 枇杷核由来ェ キスが、 リノール酸、 リノレン酸、 |3—シトステロ一ル、 /3—シトステロ一ルー 3— 0—モノグリコシド、 アミグダリン、 ベンズアルデヒド、 マンデニトリル、 安息香酸からなる群から選択される少なくとも 1種を含有することを特徴とする。 また、 本発明のフリ一ラジカル消去剤の好ましい態様としては、 枇杷核由来ェ キスが、 ）3—シトステロール及び i3—シトステロール— 3— 0—モノグリコシド を含有することを特徴とする。 In a preferred embodiment of the free radical scavenger of the present invention, the loquat nucleus-derived extract comprises a ground material obtained by pulverizing loquat nucleus, comprising ethanol, methanol, water, hexane, or a mixture thereof. It is obtained by immersing in at least one solvent selected from the group and collecting the supernatant. Further, in a preferred embodiment of the free radical scavenger of the present invention, the loquat nucleus-derived extract is selected from the group consisting of: linoleic acid, linolenic acid, | 3-sitosterol, / 3-sitosterol-3-0-monoglycoside, amygdalin It contains at least one member selected from the group consisting of benzaldehyde, mandenitrile, and benzoic acid. In a preferred embodiment of the free radical scavenger of the present invention, the loquat nucleus-derived extract comprises: 3) -sitosterol and i3-sitosterol-3-0-monoglycoside.
本発明の健康飲食品は、 枇杷核由来エキスと含有することを特徴とする。  The health food or drink of the present invention is characterized by containing a loquat core-derived extract.
また、 本発明の健康飲食品の好ましい態様としては、 枇杷核由来エキスが、 枇 杷核を粉枠して得た粉碎物を、 エタノール、 メタノール、 水、 へキサン、 又はこ れらの混合物からなる群から選択される少なくとも 1種の溶媒に浸漬して、 上清 を分取して得られることを特徴とする。  In a preferred embodiment of the health food or drink of the present invention, the loquat nucleus-derived extract is obtained by milling a loquat nucleus obtained by powdering the loquat nucleus from ethanol, methanol, water, hexane, or a mixture thereof. And immersing it in at least one solvent selected from the group consisting of:
また、 本発明の健康飲食品の好ましい態様としては、 枇杷核由来エキスが、 リ ノール酸、 リノレン酸、 /3—シトステロ一ル、 /3—シトステロール— 3— 0—モ ノグリコシド、 アミグダリン、 ベンズアルデヒド、 マンデニトリル、 安息香酸か らなる群からなる少なくとも 1種を含有することを特徴とする。  In a preferred embodiment of the health food or drink of the present invention, the loquat nucleus-derived extract comprises linoleic acid, linolenic acid, / 3-sitosterol, / 3-sitosterol-3-0-monoglycoside, amygdalin, benzaldehyde , Mandenitrile, and benzoic acid.
また、 本発明の健康飲食品の好ましい態様としては、 枇杷核由来エキスが、 β ーシトステロ一ル及び 0—シトステロ一ルー 3— 0 _モノグリコシドを含有する ことを特徴とする。  In a preferred embodiment of the health food or drink of the present invention, the loquat nucleus-derived extract contains β-cytostelol and 0-sitosterol 3-O_monoglycoside.
また、 本発明の健康飲食品の好ましい態様としては、 枇杷核由来エキスが、 さ らにリノ一ル酸を含有することを特徴とする。  In a preferred embodiment of the health food or drink of the present invention, the loquat nucleus-derived extract further contains linoleic acid.
また、 本発明の健康飲食品の好ましい態様としては、 枇杷核由来エキスが、 さ らにリノレン酸を含有することを特徴とする。  In a preferred embodiment of the health food or drink of the present invention, the loquat nucleus-derived extract further contains linolenic acid.
また、 本発明の健康飲食品の好ましい態様としては、 健康飲食品が、 健康食品 であることを特徴とする。  In a preferred embodiment of the health food or drink of the present invention, the health food or drink is a health food.
また、 本発明の健康飲食品の好ましい態様としては、 さらに、 米粉、 そば粉、 うどん粉、 小麦粉、 パン粉からなる群から選択される少なくとも 1種を含有する 事を特徴とする。 In addition, preferred embodiments of the health food and drink of the present invention further include rice flour, buckwheat flour, It contains at least one member selected from the group consisting of udon flour, flour, and bread crumbs.
また、 本発明の健康飲食品の好ましい態様としては、 本発明の健康飲食品が、 健康飲料であることを特徴とする。  In a preferred embodiment of the health food or drink of the present invention, the health food or drink of the present invention is a health drink.
また、 本発明の健康飲食品の好ましい態様としては、 さらに、 水、 ココア、 コ 一ヒー、紅茶、牛乳、酒、果実飲料、又は野菜飲料を含有することを特徵とする。 図面の簡単な説明  In addition, a preferred embodiment of the health food or drink of the present invention is characterized in that the health food or drink further contains water, cocoa, coffee, tea, milk, sake, fruit drink, or vegetable drink. BRIEF DESCRIPTION OF THE FIGURES
図 1は、 枇杷核エキスの調製方法の一例を示す図である。  FIG. 1 is a diagram showing an example of a method for preparing loquat nucleus extract.
図 2は、 各種溶媒を用いた場合の薄層クロマトグラムの結果を示す図である。 図 3 Aは、 活性酸素産生刺激剤 f ML Pを用いた場合に、 好中球からの活性酸 素（0 2—)産生に及ぼす枇杷核エキス及びその有功成分の影響を示す。 FIG. 2 is a diagram showing the results of thin-layer chromatograms when various solvents were used. Figure 3 A, when using the active oxygen production stimulator f ML P, activity oxygen from neutrophils (0 2 -) shows the effect of loquat nuclear extract and its merit components on production.
図 3 Bは、 活性酸素産生刺激剤 f ML Pを用いた場合に、 好中球からの活性酸 素（0 2— )産生に及ぼす枇杷核エキス及びその有功成分の影響を示す。 FIG. 3B shows the effect of loquat nucleus extract and its active components on the production of active oxygen (O 2 —) from neutrophils when the active oxygen production stimulant f MLP is used.
図 3 Cは、 活性酸素産生刺激剤 f ML Pを用いた場合に、 好中球からの活性酸 素（0 2 _)産生に及ぼす枇杷核エキス及びその有功成分の影響を示す。 Figure 3 C, when using the active oxygen production stimulator f ML P, shows the effect of loquat nuclear extract and its merit components on the activity oxygen (0 2 _) production from neutrophils.
図 3 Dは、 活性酸素産生刺激剤 f ML Pを用いた場合に、 好中球からの活性酸 素（0 2— )産生に及ぼす枇杷核エキス及びその有功成分の影響を示す。 Figure 3 D, when using the active oxygen production stimulator f ML P, activity oxygen from neutrophils (0 2 -) shows the effect of loquat nuclear extract and its merit components on production.
図 3 Eは、 活性酸素産生刺激剤 ί ML Pを用いた場合に、 好中球からの活性酸 素（0 2— )産生に及ぼす枇杷核エキス及びその有功成分の影響を示す。 Figure 3 E, when using the active oxygen production stimulator ί ML P, activity oxygen from neutrophils (0 2 -) shows the effect of loquat nuclear extract and its merit components on production.
図 3 Fは、 活性酸素産生刺激剤 f ML Pを用いた場合に、 好中球からの活性酸 素（0 2—)産生に及ぼす枇杷核エキス及びその有功成分の影響を示す。 Figure 3 F, when using the active oxygen production stimulator f ML P, activity oxygen from neutrophils (0 2 -) shows the effect of loquat nuclear extract and its merit components on production.
図 3 Gは、 活性酸素産生刺激剤 f ML Pを用いた場合に、 好中球からの活性酸 素（0 2— )産生に及ぼす枇杷核エキス及びその有功成分の影響を示す。 Figure 3 G, when using the active oxygen production stimulator f ML P, activity oxygen from neutrophils (0 2 -) shows the effect of loquat nuclear extract and its merit components on production.
図 3 Hは、 活性酸素産生刺激剤 f ML Pを用いた場合に、 好中球からの活性酸 素（0 2—)産生に及ぼす枇杷核エキス及びその有功成分の影響を示す。 図 3 Iは、 活性酸素産生刺激剤 f ML Pを用いた場合に、 好中球からの活性酸 素（0 2_)産生に及ぼす枇杷核エキス及びその有功成分の影響を示す。 FIG. 3H shows the effect of loquat nucleus extract and its active components on the production of active oxygen (O 2 —) from neutrophils when the active oxygen production stimulant f MLP was used. Figure 3 I, when using the active oxygen production stimulator f ML P, shows the effect of loquat nuclear extract and its merit components on the activity oxygen (0 2 _) production from neutrophils.
図 4 Aは、 活性酸素産生刺激剤 f ML Pを用いた場合に、 好中球からの活性酸 素（02— )産生に及ぼすラジカツト及びァスコルビン酸の影響を示す。 FIG. 4A shows the effect of radicat and ascorbic acid on the production of active oxygen (O 2 −) from neutrophils when the active oxygen production stimulant f MLP was used.
図 4Bは、 活性酸素産生刺激剤 f ML Pを用いた場合に、 好中球からの活性酸 素（0 2— )産生に及ぼすラジカツト及びァスコルピン酸の影響を示す。. Figure 4B, when using the active oxygen production stimulator f ML P, activity oxygen from neutrophils - shows the effect of Rajikatsuto and Asukorupin acid on the production (0 2). .
図 5Aは、 活性酸素産生刺激剤 P MAを用いた場合に、 好中球からの活性酸素 Fig.5A shows that active oxygen from neutrophils
(〇 2") 産生に及ぼす枇杷核エキス及びその有効成分の影響を示す。 (〇 2 ") The effect of loquat nucleus extract and its active ingredient on the production is shown.
図 5 Bは、 活性酸素産生刺激剤 P MAを用いた場合に、 好中球からの活性酸素 Fig. 5B shows that active oxygen from neutrophils
(〇 2— ) 産生に及ぼす枇杷核エキス及びその有効成分の影響を示す。 The effect of loquat nucleus extract and its active ingredient on (〇 2 —) production is shown.
図 5 Cは、 活性酸素産生刺激剤 P MAを用いた場合に、 好中球からの活性酸素 Fig. 5C shows active oxygen from neutrophils when the active oxygen production stimulant PMA was used.
(〇 2") 産生に及ぼす枇杷核エキス及びその有効成分の影響を示す。 (〇 2 ") The effect of loquat nucleus extract and its active ingredient on the production is shown.
図 5 Dは、 活性酸素産生刺激剤 P M Aを用いた場合に、 好中球からの活性酸素 Fig. 5D shows that reactive oxygen from neutrophils
(〇 2") 産生に及ぼす枇杷核エキス及びその有効成分の影響を示す。 (〇 2 ") The effect of loquat nucleus extract and its active ingredient on the production is shown.
図 5 Eは、 活性酸素産生刺激剤 P MAを用いた場合に、 好中球からの活性酸素 Fig. 5E shows active oxygen from neutrophils when the active oxygen production stimulating agent PMA was used.
(〇 2") 産生に及ぼす枇杷核エキス及びその有効成分の影響を示す。 (〇 2 ") The effect of loquat nucleus extract and its active ingredient on the production is shown.
図 5 Fは、 活性酸素産生刺激剤 P MAを用いた場合に、 好中球からの活性酸素 Fig.5F shows active oxygen from neutrophils when active oxygen production stimulating agent PMA was used.
(〇 2") 産生に及ぼす枇杷核エキス及びその有効成分の影響を示す。 (〇 2 ") The effect of loquat nucleus extract and its active ingredient on the production is shown.
図 5 Gは、 活性酸素産生刺激剤 PMAを用いた場合に、 好中球からの活性酸素 Fig.5G shows that active oxygen from neutrophils
(〇 2") 産生に及ぼす枇杷核エキス及びその有効成分の影響を示す。 (〇 2 ") The effect of loquat nucleus extract and its active ingredient on the production is shown.
図 5 Hは、 活性酸素産生刺激剤 P MAを用いた場合に、 好中球からの活性酸素 Fig. 5H shows the active oxygen from neutrophils when the active oxygen production stimulating agent PMA was used.
(〇 2~) 産生に及ぼす枇杷核エキス及びその有効成分の影響を示す。 2 shows the effect of loquat nucleus extract and its active ingredient on (〇2 ~) production.
図 5 Iは、 活性酸素産生刺激剤 P MAを用いた場合に、 好中球からの活性酸素 Fig. 5I shows that active oxygen from neutrophils
(〇 2~) 産生に及ぼす枇杷核エキス及びその有効成分の影響を示す。 2 shows the effect of loquat nucleus extract and its active ingredient on (〇 2 ~) production.
図 6 Aは、 活性酸素産生刺激剤 P MAを用いた場合に、 好中球からの活性酸素 Fig. 6A shows active oxygen from neutrophils when the active oxygen production stimulant PMA was used.
(〇2— ) 産生に及ぼすラジカツト及びァスコルビン酸の影響を示す。 図 6Bは、 活性酸素産生刺激剤 PMAを用いた場合に、 好中球からの活性酸素The effect of radicat and ascorbic acid on (〇 2 —) production is shown. Fig. 6B shows that active oxygen from neutrophils
(〇2— ) 産生に及ぼすラジカツト及びァスコルビン酸の影響を示す。 The effect of radicat and ascorbic acid on (〇 2 —) production is shown.
図 7 Aは、 活性酸素産生刺激剤 A Aを用いた場合に、 好中球からの活性酸素 Fig. 7A shows active oxygen from neutrophils when active oxygen production stimulant A A was used.
(02") 産生に及ぼす枇杷核エキス及びその有効成分の影響を示す。 The effect of loquat nucleus extract and its active ingredient on (0 2 ") production is shown.
図 7Bは、 活性酸素産生刺激剤 PMAを用いた場合に、 好中球からの活性酸素 Fig. 7B shows that active oxygen from neutrophils
(02~) 産生に及ぼすラジカツ卜及びァスコルビン酸の影響を示す。 2 shows the effects of radiocut and ascorbic acid on (0 2 ~) production.
図 7Cは、 活性酸素産生刺激剤 PMAを用いた場合に、 好中球からの活性酸素 Fig.7C shows that active oxygen from neutrophils
(02") 産生に及ぼすラジカツト及びァスコルビン酸の影響を示す。 (0 2 ) shows the effect of radicat and ascorbic acid on production.
図 7Dは、 活性酸素産生刺激剤 PMAを用いた塲合に、 好中球からの活性酸素 Fig. 7D shows active oxygen from neutrophils using the active oxygen production stimulant PMA.
(02") 産生に及ぼすラジカツト及びァスコルビン酸の影響を示す。 (0 2 ) shows the effect of radicat and ascorbic acid on production.
図 7Eは、 活性酸素産生刺激剤 PMAを用いた場合に、 好中球からの活性酸素 Figure 7E shows that reactive oxygen production from neutrophils
(02") 産生に及ぼすラジカツト及びァスコルビン酸の影響を示す。 (0 2 ) shows the effect of radicat and ascorbic acid on production.
図 7Fは、 活性酸素産生刺激剤 PMAを用いた場合に、 好中球からの活性酸素 Fig.7F shows that active oxygen from neutrophils
(02") 産生に及ぼすラジカツト及びァスコルビン酸の影響を示す。 (0 2 ) shows the effect of radicat and ascorbic acid on production.
図 7Gは、 活性酸素産生刺激剤 PMAを用いた場合に、 好中球からの活性酸素 Fig.7G shows that active oxygen from neutrophils
(02") 産生に及ぼすラジカツト及びァスコルビン酸の影響を示す。 (0 2 ) shows the effect of radicat and ascorbic acid on production.
図 7Hは、 活性酸素産生刺激剤 PMAを用いた場合に、 好中球からの活性酸素 Fig. 7H shows that active oxygen from neutrophils
(02") 産生に及ぼすラジカツト及びァスコルビン酸の影響を示す。 (0 2 ) shows the effect of radicat and ascorbic acid on production.
図 7 Iは、 活性酸素産生刺激剤 PMAを用いた場合に、 好中球からの活性酸素 (〇2— ) 産生に及ぼすラジカツト及びァスコルビン酸の影響を示す。 FIG. 71 shows the effect of radicat and ascorbic acid on the production of active oxygen (〇 2 ) from neutrophils when the active oxygen production stimulant PMA was used.
図 8 Aは、 活性酸素産生刺激剤 AAを用いた場合に、 好中球からの活性酸素 Fig. 8A shows that active oxygen from neutrophils
(〇2— ) 産生に及ぼすラジカツト及びァスコルビン酸の影響を示す。 The effect of radicat and ascorbic acid on (〇 2 —) production is shown.
図 8Bは、 活性酸素産生刺激剤 PMAを用いた場合に、 好中球からの活性酸素 Fig. 8B shows that active oxygen from neutrophils
(〇2— ) 産生に及ぼすラジカツト及びァスコルビン酸の影響を示す。 The effect of radicat and ascorbic acid on (〇 2 —) production is shown.
図 9は、 正常家兎の生物学的検査値を（GOT、 GPT、 総コレステロール値、 LDLコレステロール値、 HDLコレステロール値、 トリダリセライド値）を調べ た結果を示す図である。 Figure 9 shows the biological test values (GOT, GPT, total cholesterol, LDL cholesterol, HDL cholesterol, and tridalicelide) of normal rabbits. FIG.
図 1 0は、 1 %コレステロ一ノレ含有飼料投与家兎の生化学的検査値を調べた結 果を示す図である。  FIG. 10 is a diagram showing the results of examining the biochemical test values of rabbits to which a feed containing 1% cholesterol monocle was administered.
図 1 1は、 高脂血症家兎の生化学的検査値を調べた結果を示す図である。 図 1 2は、 高脂血症家兎の生化学的検査値を調べた結果を示す図である。 図 1 3は、正常家鬼の生物学的検査値を（GO T、 G P T、総コレステロール値、 FIG. 11 is a diagram showing the results of examining biochemical test values of hyperlipidemic rabbits. FIG. 12 is a diagram showing the results of examining biochemical laboratory values of hyperlipidemic rabbits. Figure 13 shows the biological test values (GOT, GPT, total cholesterol,
L D Lコレステロール値、 HD Lコレステロール値、 トリダリセライド値）を調べ た結果を示す図である。 It is a figure which shows the result of having investigated LDL cholesterol level, HD L cholesterol level, and tridalicelide level.
図 1 4は、 1 %コレステロール含有飼料投与家兎の生化学的検査値を調べた結 果を示す図である。  FIG. 14 is a diagram showing the results of examination of biochemical test values of rabbits to which a diet containing 1% cholesterol was administered.
図 1 5は、 高脂血症家兎の生化学的検査値を調べた結果を示す図である。 図 1 6は、 高脂血症家兎の生化学的検査値を調べた結果を示す図である。 発明を実施するための最良の形態  FIG. 15 shows the results of examining the biochemical test values of hyperlipidemic rabbits. FIG. 16 is a diagram showing the results of examining the biochemical test values of hyperlipidemic rabbits. BEST MODE FOR CARRYING OUT THE INVENTION
本発明のフリーラジカル消去剤は、枇杷核エキスを含有する。枇杷核エキスは、 枇杷の種子由来のエキスである。 本発明に適用する枇杷核エキスは、 枇杷の種子 由来である限り、 全ての枇杷核エキスを対象とする。  The free radical scavenger of the present invention contains loquat core extract. Loquat core extract is an extract derived from loquat seeds. The loquat nucleus extract applied to the present invention covers all loquat nucleus extracts as long as they are derived from loquat seeds.
ここで、 フリーラジカルとは、 遊離基とも呼ばれ、 広く不対電子を持つ原子団 を意味する。 一般的に、 フリーラジカルを持つ分子及びフリーラジカルを生成す る物質は、 反応性に富み、 細胞には有害である。 D NAについて考えれば、 D N Aを損傷させ、 突然変異を誘発させる。 このようなフリーラジカルを、 本発明に おいては効率よく除去することが可能となる。  Here, a free radical is also called a free radical and means an atomic group that has a wide unpaired electron. In general, molecules with free radicals and substances that produce free radicals are highly reactive and harmful to cells. Given DNA, it damages DNA and induces mutations. Such free radicals can be efficiently removed in the present invention.
〔枇杷核エキスの調製方法〕  (Method for preparing loquat nucleus extract)
まず、図 1を用いて、杜 I：杷核エキスの調製方法について一例を挙げて説明する。 図 1は、 枇杷核エキスの調製方法の一例を示す図である。 枇杷核は、 必要に応じ て洗浄し、 乾燥する。 乾燥は十分に行なうのが好ましい。 後の粉碎を均質に行な うためである。 First, a method for preparing forest I: Hakone extract will be described with reference to FIG. FIG. 1 is a diagram showing an example of a method for preparing loquat nucleus extract. Loquat nuclei are washed and dried if necessary. Drying is preferably performed sufficiently. The subsequent grinding is carried out homogeneously. To do so.
次に、 枇杷核を粉碎する。 粉砕の方法は特に限定されず、 ポールミル、 ハンマ 一ミル、 ローラーミル、 ロッドミル、 サンプルミル、 スタンプミル、 デイスイン テグレーター、 乳鉢、 冷却装置付きプレンダ一などの公知の粉砕機を用いること ができる。 なお、 粉砕時における発熱により、 枇杷核組成物の分解等が発生する ことも考えられることより、 冷却装置付きプレンダ一が好ましい。  Next, the loquat nucleus is ground. The method of pulverization is not particularly limited, and a known pulverizer such as a pole mill, a hammer mill, a roller mill, a rod mill, a sample mill, a stamp mill, a date integrator, a mortar, and a blender with a cooling device can be used. It is to be noted that since the loquat nucleus composition may be decomposed due to heat generated during the pulverization, a blender with a cooling device is preferable.
枇杷核を粉砕し粉砕物を得た後、 各種溶媒に前記粉碎物を浸漬する。 この場合 の溶媒は、 特に限定されず、 所望とする効果に対応して適宜溶媒を設定すること ができる。溶媒としては、 エタノール、 メタノール、水、へキサン、酢酸ェチル、 クロ口ホルム、 アセトンなどの極性、 非極性溶媒を問わず挙げることができる。 細胞膜透過性が高い枇杷核エキスを得ることができるという観点から、 好ましく は、 メタノール、 エタノール、 水等である。  After crushing loquat nuclei to obtain a crushed product, the crushed material is immersed in various solvents. The solvent in this case is not particularly limited, and a solvent can be appropriately set according to a desired effect. Examples of the solvent include any polar or non-polar solvent such as ethanol, methanol, water, hexane, ethyl acetate, chloroform, and acetone. From the viewpoint that loquat nucleus extract having high cell membrane permeability can be obtained, methanol, ethanol, water and the like are preferable.
浸漬は、 緩やかな攪拌下で行なうことができる。 各種溶媒に前記粉碎物を浸漬 して各種溶液を得る。 各種溶液について， 溶液の状態に応じて攪拌を行い、 場合 によりそのまま溶液を放置しても良い。 攪拌する場合には、 特に限定されないが The immersion can be performed under gentle stirring. The above-mentioned crushed material is immersed in various solvents to obtain various solutions. Various solutions may be stirred according to the state of the solution, and the solution may be left as it is in some cases. When stirring, there is no particular limitation.
5〜1 0日間攪拌を持続させることができる. Stirring can be sustained for 5-10 days.
その後、 上清を分取して蒸発乾固し、 枇杷核エキス末を得ることが出来る。 蒸 発乾固は、 エバポレー夕を用いて、 5 5 °C〜8 0 °Cの温浴上で行なうことができ る。  Thereafter, the supernatant is collected and evaporated to dryness to obtain loquat nucleus extract powder. Evaporation to dryness can be performed in a warm bath at 55 ° C to 80 ° C using an evaporator.
〔枇杷核エキスの成分〕  [Ingredients of loquat core extract]
枇杷核中に含まれる成分は、 枇杷核を極性の異なる溶媒を用いて抽出すること により、 その極性により振り分けられる。 したがって、 使用した溶媒により、 枇 杷核エキスの成分の種類及び含有量は異なる。  Components contained in loquat nuclei are sorted by their polarities by extracting loquat nuclei with solvents of different polarities. Therefore, the type and content of the components of the loquat core extract differ depending on the solvent used.
図 2は、 各種枇杷核エキスの成分を調べるために、 各種溶媒を用いて抽出した 枇杷核エキスの薄層クロマトグラム （T L C) を示す。  Figure 2 shows thin layer chromatograms (TLC) of loquat nucleus extract extracted using various solvents to examine the components of various loquat nucleus extracts.
この薄層クロマトグラムによれば、 溶媒が水であるエキス （以下、 水エキスと いう。） は、 原点にのみスポットが認められ、 したがって、 タンパ夕質、 糖類、 ァ ミグダリン等の極性の高い化合物を含有すると考えられる。 According to this thin-layer chromatogram, the extract in which the solvent is water (hereinafter referred to as the water extract) Say. ) Shows a spot only at the origin, and is considered to contain highly polar compounds such as tampa, sugars and amigdarin.
また、 溶媒が 7 0 %エタノールであるエキス （以下、 7 0 %エタノールエキス という。）、及び溶媒がメタノールであるエキス（以下、メタノールエキスという。） は、 薄層クロマトグラムにおいて原点のスポットが水エキスに比較して小さく、 タンパク質、 糖類、 アミグダリン等の極性の高い化合物が少ないと考えられる。 7 0 %エタノールエキス及びメタノールエキスにおいては、 薄層クロマトグラム にて: f値が 0. 6 3を示す化合物はリノレン酸、 R f値が 0 . 5 3を示す化合 物は、 ；6—シトステロール、 R f値が 0 . 4 1を示す化合物はリノール酸、 R f 値が 0 . 2 5を示す化合物は /3—シトステロ一ル— 3— 0—モノグリコシドであ ることが、 構造解析により判明したことにより、 これらの化合物を少なくとも含 む。  Extracts in which the solvent is 70% ethanol (hereinafter referred to as 70% ethanol extract) and extracts in which the solvent is methanol (hereinafter referred to as methanol extract) have water spots at the origin in the thin-layer chromatogram. It is thought that there are few highly polar compounds such as proteins, sugars, and amygdalin, which are smaller than extracts. For 70% ethanol extract and methanol extract, thin layer chromatograms show: compounds with f-value 0.63 are linolenic acid, compounds with R f 0.53 are: 6-sitosterol According to structural analysis, the compound having an R f value of 0.41 was linoleic acid, and the compound having an R f value of 0.25 was / 3-cytositol-3-0-monoglycoside. According to the findings, these compounds are included at least.
また、 溶媒がへキサンであるエキス （以下、 へキサンエキス） は、 薄層クロマ 卜グラムにて R f値が 0 . 7 1≥の化合物が確認され、 極性の低い化合物を多く 含む  Extracts with hexane as the solvent (hereinafter referred to as hexane extract) show compounds with an R f value of 0.71 ≥ in thin-layer chromatograms and contain many low-polarity compounds.
と考えられる。 it is conceivable that.
更に、 磁気共鳴法（NMR)、 質量分析法 （M S ) 紫外吸収スぺクトル法 （I R) 等を用いて成分分析を行った。 その結果、 少なくともリノール酸、 リノレン酸、 ]3—シトステロール、 j6—シトステロール— 3— 0—モノグリコシド、 アミグダ リン、 ベンズアルデヒド、 マンデニトリル及び安息香酸の 8種類の有効成分及び プロリン類似化合物が確認された。 枇杷核エキスより単離した化合物を以下の表 1に示す  Further, the components were analyzed using a magnetic resonance method (NMR), a mass spectrometry method (MS), an ultraviolet absorption spectrum method (IR), and the like. As a result, at least eight active ingredients, linoleic acid, linolenic acid,] 3-sitosterol, j6-sitosterol-3-0-monoglycoside, amygdalin, benzaldehyde, mandenitrile, and benzoic acid, and a proline-like compound were confirmed. . The compounds isolated from Loquat Nucleus Extract are shown in Table 1 below.
表 1 枇杷エキス中より単離した化合物 table 1 Compound isolated from loquat extract
No. 化合物名 構造式 主な薬理活性 No. Compound name Structural formula Main pharmacological activity
(CH2)6 COOH コレス卜ローレ(CH 2 ) 6 COOH
1 リノール酸
Figure imgf000011_0001
1 Linoleic acid
Figure imgf000011_0001
溶解作用  Dissolving action
コレストロールCholesterol
2 リノレン酸 H3Cヽ /= ノ（CH2)6 C00H 2 Linolenic acid H 3 C ヽ / = NO (CH 2 ) 6 C00H
溶解作用  Dissolving action
高脂血症 Hyperlipidemia
3 ]3 -シトステロール  3] 3-sitosterol
改善作用
Figure imgf000011_0002
Improvement action
Figure imgf000011_0002
Figure imgf000012_0001
Figure imgf000012_0001
鎮咳去痰Antitussive expectorant
7 マンデニトリル 7 Mandenitrile
H0'TCN 作用 H0'T CN action
鎮痛 Pain relief
8 安息香酸  8 Benzoic acid
尿路防腐剤
Figure imgf000013_0001
Urinary preservatives
Figure imgf000013_0001
したがって、 本発明の枇杷核エキスを含有する医薬組成物は、 上記 8種類の有 効成分のうち少なくとも 1種を含む。 本発明の枇杷核エキスを含有する医薬組成 物は、 好ましくは、 ]8—シトステロール、 及び j8—シトステロール一 3— 0—モ ノグリコシドを含有する。 ]3—シトステロール及び |3—シトステロール一 3— 0 —モノグリコシドは、 フリーラジカル消去作用及び高脂血症改善作用を直接的に 有するからである。 本発明の枇杷核エキスを含有する医薬組成物は、 ]3シトステ 口一ル及び) 3 -シトステロ一ルー 3— 0—モノダリコシドのほか、 さらにリノ一 ル酸、 及び Z又はリノレン酸を含むことが好ましい。 リノール酸及びリノレン酸 は、 コレステロール溶解作用を有するからである。 Therefore, the pharmaceutical composition containing the loquat nucleus extract of the present invention contains at least one of the above eight active ingredients. The pharmaceutical composition containing loquat nucleus extract of the present invention preferably contains] 8-sitosterol and j8-sitosterol-13-0-monoglycoside. ] 3-sitosterol and | 3-sitosterol-1-3-0-monoglycoside have a free radical scavenging action and a hyperlipidemia-improving action directly. The pharmaceutical composition containing the loquat nucleus extract of the present invention may contain, in addition to] 3 sitosterone and 3) -sitosterol-3-0-monodalicoside, linoleic acid, and Z or linolenic acid. preferable. Linoleic acid and linolenic acid have a cholesterol dissolving action.
〔有効量〕 [Effective amount]
本発明によるフリーラジカル消去剤は、 有効的な量の枇杷核エキス、 及び適当 な投与形態の形で調製される。  The free radical scavenger according to the invention is prepared in an effective amount of loquat nucleus extract and in a suitable dosage form.
本発明のフリーラジカル消去剤における枇杷核エキスの投与量は、 投与対象患 者の病態及びその重篤度、 投薬形態、 選択した投与経路及び 1日当たりの投与回 数等により変更することができる。  The dose of loquat nucleus extract in the free radical scavenger of the present invention can be changed depending on the condition and severity of the subject to be administered, the dosage form, the selected administration route, the number of administrations per day, and the like.
本発明のフリーラジカル消去剤における枇杷核エキスの投与量は、 ラットにお いては少なくとも 3 7 5mg/kg/日となる量であり、ヒトにおいては感受性の相違等 により、 更に低い量であることが好ましい。  The dose of loquat nucleus extract in the free radical scavenger of the present invention should be at least 375 mg / kg / day in rats, and even lower in humans due to differences in sensitivity. Is preferred.
また、投与には、経口剤（タブレツト、カプセル、被膜タブレツト、顆粒、溶液、 シロップ)、 直腸投与用坐剤、 注射剤等を用いることができる。 投与対象患者は、 慢性疾患の場合が多ぐ 長期投与の必要性、 運用の容易性という観点から、 好ま しくは、 経口剤である。  For administration, oral preparations (tablets, capsules, coated tablets, granules, solutions, syrups), suppositories for rectal administration, injections and the like can be used. Patients to be administered are often oral drugs from the viewpoint of the necessity of long-term administration and the easiness of operation, which are often chronic diseases.
投与には、 従来の他の成分、 例えば、 安定保存剤、 甘味料、 着色剤、 芳香剤な どを含むことができる。  Administration can include other conventional ingredients such as stable preservatives, sweeteners, coloring agents and flavoring agents.
〔急性毒性試験〕 Carter. J.H. らは、 枇杷核エキスの含有成分であるアミグダリンの毒性 について検討し、 ラットにおけるアミグダリンの致死量は、 600mgであると報告 している (Carter. J.H. Mclefferty, Μ. Α·， GoldOan, F.， Biochem. Pharmacol. 29,301 ( 1980))。 (Acute toxicity test) JH et al. Examined the toxicity of amygdalin, a component of loquat nucleus extract, and reported that the lethal dose of amygdalin in rats was 600 mg (Carter. JH Mclefferty, Μ. Α ·, GoldOan, F., Biochem. Pharmacol. 29,301 (1980)).
枇杷核エキスの致死量は、 そのアミグダリン含量 （7. 4%) より、 8060 mgZkgと推定できる。  The lethal dose of loquat nucleus extract can be estimated to be 8060 mgZkg based on its amygdalin content (7.4%).
〔枇杷核エキスの調製方法〕  (Method for preparing loquat nucleus extract)
枇杷核は、 高知県内にて採取した枇杷の種子を日干しにより十分に乾燥させた ものを用いた。 枇杷核エキスは、 冷却装置付プレンダ一にて粉碎 （1， 000 r pm) した枇杷核 （1, 050 g) を倍量の 70%エタノール （2， 1000m 1) に浸漬、 攪拌器にて常時攪拌 （300 r pm) し、 浸漬後 7日に上清を分取 し、 エバポレー夕にて 70°Cの温浴で蒸発乾固し、 製した。  The loquat core used was loquat seeds collected in Kochi Prefecture and dried sufficiently by sun drying. Loquat nucleus extract is obtained by immersing loquat nucleus (1,050 g) ground (1,000 rpm) in a blender with a cooling device in twice the volume of 70% ethanol (2,1000 m1), and constantly using a stirrer. After stirring (300 rpm), the supernatant was collected on the 7th day after the immersion, and evaporated to dryness in a hot bath at 70 ° C in an evaporator to produce the product.
なお、 枇杷核エキス末の収量は 10. 4g (抽出率 1. 0%) であった。 〔成分分析〕  The yield of loquat core extract powder was 10.4 g (extraction rate: 1.0%). (Component analysis)
枇杷核エキスのフリーラジカル消去作用を検討する為に、 まず、 枇杷核エキス に含有される有効成分について調べた。 即ち、 枇杷核エキスを調製 (図 1)し、 溶 媒間分画法、 シリカゲルカラムクロマトグラフィー、 分子ふるいカラムクロマト グラフィー及び薄層クロマトグラフィー （TLC) を用いて成分分画を行い、 8 種類の成分を単離した。これらの化合物について高速液体クロマトグラフィー（H PLC), TLC等を用いた標品との比較、 及び核磁気共鳴法 （NMR)、 質量分 析法（MS)、 赤外吸収スぺクトル法（I R)等を用いた機器分析により構造決定 を行った。 その結果、 枇杷核エキスは、 /3—シトステロール、 ]3—シトステロ一 ルー 3— 0—モノグリコシド、 リノール酸、 リノレン酸、 アミグダリン、 ベンズ アルデヒド、 マンデニトリル及び安息香酸を含有することが判明した。 以下に、 その同定結果を示す。 測定条件等については、 以下の通りである。  In order to examine the free radical scavenging effect of loquat nucleus extract, first, the active ingredients contained in loquat nucleus extract were examined. That is, loquat nucleus extract was prepared (Fig. 1), and the components were fractionated using solvent separation method, silica gel column chromatography, molecular sieve column chromatography, and thin layer chromatography (TLC), and eight kinds of components were separated. The components were isolated. Comparison of these compounds with samples using high performance liquid chromatography (HPLC), TLC, etc., and nuclear magnetic resonance (NMR), mass spectrometry (MS), infrared absorption spectrum (IR) ) Was determined by instrumental analysis. As a result, it was found that the loquat core extract contained / 3-sitosterol,] 3-sitosterol 3-0-monoglycoside, linoleic acid, linolenic acid, amygdalin, benzaldehyde, mandenitrile and benzoic acid. The results of the identification are shown below. The measurement conditions are as follows.
〔|S—シトステロール〕 i3—シトステロールは、 枇杷核エキスより単離し、 TLC，EI- MS， 1HNMR(CDC1 2)、 スぺクトルにおいて標品のデータを比較同定した。 [| S—Sitosterol] i3-sitosterol was isolated from loquat nucleus extract, and its data were compared and identified by TLC, EI-MS, 1HNMR (CDC12), and spectrum.
TLC展開溶媒 CHC13: MeOH: H20 (8： 2 :0.2) R f値 0.7 1 HR-EI-MS C2g¾。0として、 理論値 14.378, 実測値 414.387 1HNMR (ピリジン— d5) 141.0 (s、 10 12 1.9 (d、 10 102.6 (d、 1C)、 56.9 (d、 1C)、 56.4 (d、 1C)、 50.5 (d、 1C)、 46.2 (d、 1C)、 42.6 (s、 10 40.0 (t、 1C)、 39.4 (t、 1C)、 370 (t、 1C)、 370 (s、 1C)、 36.4 (d、 1 C) 34.3 (t、 1C)、 32.24 (t、 1C)、 32.15 (d、 1C)、 32.15 (d、 1C)、 30.3 (t、 1 C) 29.6 (d、 1 C)、 28.6 (t、 1C)、 26.6 (t、 1C)、 24.6 (t、 IC)、 23.5 (1、 1 C)、 （t、 1C)、 20.0 (q、 10, 19.5 (q、 1C)、 19.3 (q、 1C)、 1 9. 1 (q iC)、 12.2 (q、 1C)、 12 · 0 (q、 1 C) TLC developing solvent CHC13: MeOH: H 2 0 ( 8: 2: 0.2) R f value 0.7 1 HR-EI-MS C 2g ¾. Assuming 0, theoretical value 14.378, actual value 414.387 1H NMR (pyridine-d5) 141.0 (s, 10 12 1.9 (d, 10 102.6 (d, 1C), 56.9 (d, 1C), 56.4 (d, 1C), 50.5 ( d, 1C), 46.2 (d, 1C), 42.6 (s, 10 40.0 (t, 1C), 39.4 (t, 1C), 370 (t, 1C), 370 (s, 1C), 36.4 (d, 1 C) 34.3 (t, 1C), 32.24 (t, 1C), 32.15 (d, 1C), 32.15 (d, 1C), 30.3 (t, 1C) 29.6 (d, 1C), 28.6 (t, 1C ), 26.6 (t, 1C), 24.6 (t, IC), 23.5 (1, 1C), (t, 1C), 20.0 (q, 10, 19.5 (q, 1C), 19.3 (q, 1C), 19.1 (q iC), 12.2 (q, 1C), 12.0 (q, 1C)
IR (KBr, cm - 1) 3420  IR (KBr, cm-1) 3420
;8—シトステロール一 3— 0—モノグリコシド  ; 8-sitosterol 3-0-monoglycoside
/3—シトスチロール一 3— 0—モノグリコシドは、 枇杷核エキスより単離、 H NMR (び、 CDC13) 及び 13C— NMR (び、 ピリジン - d5) スペクトルにおい て標品のデータと比較同定した。  1 / 3-0-Glycoside was isolated from loquat nucleus extract and identified by comparison with the standard data in 1 H NMR (bi, CDC13) and 13 C-NMR (bi, pyridine-d5) spectra. .
1HNMR (δ、 ピリジン- d5) 5.6-3.8 (m) 2.8-0.6(m)  1H NMR (δ, pyridine-d5) 5.6-3.8 (m) 2.8-0.6 (m)
13C- NMR (<5、 ピリジン- d5) 141.0 (s、 1C)、 121.9 (d、 1C)、 102.6 (d、 10 78.6 (d、 1C)、 78.4 (d、 1C)、 78.3 (d、 1C)、 75.3 (d、 1C)、 71.8 (d、 1C)、 62.9 (t, 1C)、 56.9 (d、 1C)、 56.4 (d、 1C)、 50.5 (d、 1C)、 46.2 (d、 1C)、 42.6 (s、 1C)、 40.0 (t、 1C)、 39.4 (t、 1C)、 37.6 (t、 1C)、 370 (s、 1C)、 さ 6.4 (d、 1C)、 34.3 (t、 1C)、 32.24 (t、 1C)、 32.15 (d、 1C)、 30.3 (t 1C)、 29.6 (d、 1C)、 28.6 (t、 1C)、 26.る （1、 IC)、 24.6 (t、 1C)、 23.5 〈t、 1C)、 2 1.4 (t、 1C)、 20.0 (Q、 1C)、 19.5 (q、 IC)、 19.3 (Q、 1C)、 19.1 (q、 1C)、 12.0 (q、 1C)、 12.9 (q、 10 13C-NMR (<5, pyridine-d5) 141.0 (s, 1C), 121.9 (d, 1C), 102.6 (d, 10 78.6 (d, 1C), 78.4 (d, 1C), 78.3 (d, 1C) , 75.3 (d, 1C), 71.8 (d, 1C), 62.9 (t, 1C), 56.9 (d, 1C), 56.4 (d, 1C), 50.5 (d, 1C), 46.2 (d, 1C), 42.6 (s, 1C), 40.0 (t, 1C), 39.4 (t, 1C), 37.6 (t, 1C), 370 (s, 1C), 6.4 (d, 1C), 34.3 (t, 1C), 32.24 (t, 1C), 32.15 (d, 1C), 30.3 (t 1C), 29.6 (d, 1C), 28.6 (t, 1C), 26.ru (1, IC), 24.6 (t, 1C), 23.5 <t, 1C), 21.4 (t, 1C), 20.0 (Q, 1C), 19.5 (q, IC), 19.3 (Q, 1C), 19.1 (q, 1C), 12.0 (q, 1C), 12.9 (q, 10
安息香酸は、 枇杷核エキスより単離し、 1R， HR-EI-MS, 1HNMR(CDC13) 及び HR- EI - MSスぺクトルにおいて標品のデータと比較同定した。 Benzoic acid was isolated from loquat nucleus extract and identified by comparison with standard data in 1R, HR-EI-MS, 1HNMR (CDC13) and HR-EI-MS spectra.
IR (KBr、 cm- 1) 3432、 3072、 1687、 1602  IR (KBr, cm-1) 3432, 3072, 1687, 1602
1HNMR (δ, CDC 13) 8.15-8.10 (m, 2H) 7.66- 7.60 (m. 1 H) 7.52-7. 6 (m, 2H) 13C-NMR (δ, CDC 13) 171.2 (s、 1C)、 133.8 (d、 1C)、 130.2 (d、 2C)、 129.2 (s、 10, 128.5 1H NMR (δ, CDC 13) 8.15-8.10 (m, 2H) 7.66- 7.60 (m.1H) 7.52-7.6 (m, 2H) 13C-NMR (δ, CDC 13) 171.2 (s, 1C), 133.8 (d, 1C), 130.2 (d, 2C), 129.2 (s, 10, 128.5
(d、 20 (d, 20
HR-EI-MS C7H602として、 理論値 122.043、 実測値 122.036 HR-EI-MS C 7 H as 6 0 2, the theoretical value 122.043, found 122.036
〔リノール酸及びリノレン酸〕 (Linoleic acid and linolenic acid)
リノ一ル酸及びリノレン酸は、 枇杷核エキスを HP LCに付し、 標品と比較同定 した。 枇杷核エキスの H PLCクロマトにおいて、 リノール酸及びリノレン酸の 標品と同一のリテンションタイムを示すピークが認められた。 リノ一ル酸及びリノレン酸の H PLCの測定条件 日立 D 600型高速液体クロマトグラフ For linoleic acid and linolenic acid, loquat nucleus extract was subjected to HP LC, and compared with the authentic sample. In the HPLC of loquat nucleus extract, a peak showing the same retention time as the linoleic acid and linolenic acid samples was observed. HPLC measurement conditions for linoleic acid and linolenic acid Hitachi D600 High Performance Liquid Chromatograph
検出 240 nm Detection 240 nm
カラム YMC Pack ODS-AQ 150X6.6imn I.D. Column YMC Pack ODS-AQ 150X6.6imn I.D.
測定温度 30°C Measurement temperature 30 ° C
移動相 ァセトニトリル：水：酢酸 （8 : 2 : 0. 1) Mobile phase acetonitrile: water: acetic acid (8: 2: 0.1)
流速 0.7ml/min a U 9. 98分 Flow rate 0.7ml / min a U 9.98 minutes
リノール酸 14. 09分  Linoleic acid 14.09 minutes
〔アミグダリン、 ベンズアルデヒド及びマンデニトリル〕 (Amygdalin, benzaldehyde and mandenitrile)
アミグダリン、 ベンズアルデヒド及びマンデニトリルは、 枇杷核エキスを HP LCに付し、 標品と比較同定した。  For amygdalin, benzaldehyde and mandenitrile, loquat nucleus extract was subjected to HP LC and identified by comparison with the standard.
枇杷核エキスの H PLCクロマトにおいて、 アミグダリン、 ベンズアルデヒド 及びマンデニトリルの標品と同一のリテンションタイムを示すピークが認められ た。 アミグダリンの H P L Cの測定条件  In the HPLC of loquat nucleus extract, a peak showing the same retention time as the amygdalin, benzaldehyde and mandenitrile preparations was observed. Measurement conditions for amygdalin HPLC
日立 D 600型高速液体クロマトグラフ  Hitachi D 600 High Performance Liquid Chromatograph
検出 UV242 nm Detection UV242 nm
カラム YMC Pack ODS-AQ 150X6.6mm I.D. Column YMC Pack ODS-AQ 150X6.6mm I.D.
測定温度 30°C Measurement temperature 30 ° C
移動相 ァセ卜二卜リル：水 （1 ： 4) Mobile phase acetate: water (1: 4)
1 m 1 / ί η  1 m 1 / ί η
リテンションタイム： 5. 15分 ベンズアルデヒドの H PLCの測定条件 Retention time: 5.15 minutes HPLC conditions for benzaldehyde
日立 D 600型高速液体クロマトグラフ  Hitachi D 600 High Performance Liquid Chromatograph
検出 UV280 nm Detection UV280 nm
カラム YMC Pack ODS-AQ 150 X 6.6mm I.D. Column YMC Pack ODS-AQ 150 X 6.6mm I.D.
測定温度 30。C Measurement temperature 30. C
移動相 ァセトニトリル： 2%酢酸 （2 ： 1) 流速 1 m 1 / i n Mobile phase acetonitrile: 2% acetic acid (2: 1) Flow rate 1 m 1 / in
リテンションタイム： 4. 0 5分 マンデニトリルの H P L Cの測定条件 Retention time: 4.0 5 minutes Measurement conditions of HPLC of mandenitrile
装置 日立 D 6 0 0型高速液体クロマトグラフ Equipment Hitachi D600 High Performance Liquid Chromatograph
検出 2 8 0 n m Detection 2 8 0 n m
カラム YMC Pack 0DS-AQ 1 50 X 6. 6mm I. D. Column YMC Pack 0DS-AQ 1 50 X 6.6mm I.D.
測定温度 3 0 °C Measurement temperature 30 ° C
移動相 メタノール：水 （7 ： 3 ) Mobile phase methanol: water (7: 3)
流速 1 m 1 /m i n Flow rate 1 m 1 / min
リテンションタイム： 4. 9 8分 次に、 本発明の健康食品又は健康飲料について説明する。 本発明の健康食品又 は健康飲料は、 枇杷核エキスを含有する。 枇杷核エキスは、 枇杷の種子由来のェ キスである。 本発明に適用する枇杷核エキスは、 枇杷の種子由来である限り、 全 ての枇杷核エキスを対象とする。 Retention time: 4.98 minutes Next, the health food or health drink of the present invention will be described. The health food or health drink of the present invention contains loquat core extract. Loquat core extract is an extract derived from loquat seeds. The loquat nucleus extract applied to the present invention covers all loquat nucleus extracts as long as they are derived from loquat seeds.
また、 健康食品又は健康飲料の用語は、 特に限定されず、 広く枇杷核エキスを 含有する食品又は飲料を意味するものである. 本発明の枇杷核エキスをそのまま 食用とすることも可能であるが、 自然に食用できるように、 後述するような他の 天然素材と混合して食用することもできる。  The term healthy food or health drink is not particularly limited, and broadly means a food or beverage containing loquat nucleus extract. The loquat nucleus extract of the present invention can be used as it is. However, it can be mixed with other natural ingredients as described below for edible use.
次に、 本発明の健康食品及び健康飲料の食用方法について説明する。 枇杷核ェ キスの調整方法は、 以下の通りである。 上述のようにして得られた枇杷核エキス 由来の粉末 1〜1 0 O mg程度を、水 1 0 - 1 0 0 m 1に溶かして食用することが できる。 これを 1 日 2〜3回服用することができる。  Next, the edible method of the health food and the health beverage of the present invention will be described. The method for adjusting loquat nuclei is as follows. About 1 to 10 mg of the powder derived from loquat nucleus extract obtained as described above can be dissolved in 10 to 100 ml of water and eaten. You can take this 2-3 times a day.
枇杷披エキスを含有する健康食品は、 食事の代用、 間食として食用することも できる。 この場合、 前記枇杷核エキス由来の粉末を米粉、 そば粉、 うどん粉、 小 麦粉、 又はパン粉から選択される少なくとも 1種とを調合して通常の料理に利用 することもできる。 また、 これらを洋菓子、 和菓子、 菓子、 うどん、 そば、 パス 夕、 中華そば、 パン等に加工してさらに食べやすくしても良い。 これらの料理方 法は常法に従う。 The health food containing loquat extract can be used as a meal substitute or snack. In this case, the powder derived from the loquat core extract is rice flour, buckwheat flour, udon flour, Wheat flour or at least one selected from bread crumbs can be mixed and used for ordinary cooking. These may be processed into Western confectionery, Japanese confectionery, confectionery, udon, buckwheat noodles, pasta, Chinese noodles, bread, etc. to make them even easier to eat. These cooking methods follow common law.
また、 健康食品の状態は、 固体状（粉末、 顆粒状)、 ゼリー状、 液体、 懸濁状の いずれでもよい。 また、 健康食品、 又は健康飲料には、 甘味料、 酸味料、 ビタミ ン剤、 ドリンク剤などに含まれる各種成分等を含まれていても良い。  The health food may be in a solid (powder, granule), jelly, liquid, or suspension form. In addition, the health food or the health drink may contain various components contained in a sweetener, an acidulant, a vitamin, a drink and the like.
また健康飲料の場合には、 さらに水、 ココア、 コーヒー、 紅茶、 牛乳、 酒、 果 実飲料、 又は野菜飲料からなる群から選択される少なくとも 1種を含有してもよ い。 上述したように得られた枇杷核エキス由来の粉末 1〜1 0 O m g程度を、 水 1 0〜 1 0 O m 1に溶かして食用する場合に、 さらに水、 ココア等に溶かして飲 用することができる。  In the case of health drinks, it may further contain at least one selected from the group consisting of water, cocoa, coffee, tea, milk, alcohol, fruit drinks, and vegetable drinks. When about 1 to 10 mg of the powder derived from loquat nucleus extract obtained as described above is dissolved in 10 to 10 Om1 of water for consumption, it is further dissolved in water, cocoa, etc. for drinking. be able to.
いずれにしても、 本発明の健康食品又は健康飲料の飲食方法は自由であり、 特 に限定されない. したがって、 食品素材を活かした飲食方法であれば、 本発明の 効果を期待できる。 よって、 発明の実施態様、 及び以下の実施例において記述し た飲食方法は、 単に説明するために用いた本発明の一例にすぎず限定的な解釈と されるべきものではない。  In any case, the method of eating and drinking the health food or the healthy beverage of the present invention is free and not particularly limited. Therefore, the effects of the present invention can be expected as long as the eating and drinking method utilizes food materials. Therefore, the embodiments of the invention and the eating and drinking methods described in the following examples are merely examples of the present invention used for explanation, and should not be construed as limiting.
〔実施例〕  〔Example〕
以下では本発明の実施例を説明するが、 本発明は、 下記の実施例に限定して解 釈されるものではない。  Hereinafter, embodiments of the present invention will be described, but the present invention is not limited to the following embodiments.
実施例 1 Example 1
まず、活性酸素（02 _)消去能に対する枇杷核エキス及びその有効成分の影響に ついて調べた。 First, it was examined about the effect of loquat nuclear extract and its active ingredient to active oxygen (0 2 _) scavenging ability.
好中球は、 ラット末梢血より濃度勾配遠心法により分離した。 分離した好中球 に、 活性酸素産生刺激剤 FML P (N- f ormy卜 methi ony卜 l eucy卜  Neutrophils were separated from rat peripheral blood by concentration gradient centrifugation. The isolated neutrophils are treated with the active oxygen production stimulant FML P (N-formy methiony
phenyl alanine (N—フオルミル—メチォニル—ロイシルーフエ二ルァラ二 ン)）及び各種濃度のエキスあるいは有効成分を投与し、 産生する活性酸素量（02 ―)をチトクローム c還元法で測定した。 対照例として、 市販医薬品 （ラジカット 及びァスコルビン酸） を用いて同様に試験を行なつた。 phenyl alanine (N-formyl-methionyl-leucylofenilani Was measured) in cytochrome c reduction method - down)) and administered extract or active ingredients of various concentrations, the amount of active oxygen produced (0 2. As a control, the same test was carried out using commercially available drugs (Radicut and ascorbic acid).
なお、 活性酸素消去能は、 枇杷核エキス及びその有効成分の投与群と無投与群 との比により求めた活性酸素産生抑制率及び活性酸素除去率で表した。  In addition, the active oxygen scavenging ability was represented by an active oxygen production suppression rate and an active oxygen removal rate determined by a ratio between a group administered with loquat nucleus extract and its active ingredient and a group not administered.
結果を、 図 3(a)〜（i)に示す。 図 3は、 好中球からの活性酸素（02 -)産生に 及ぼす枇杷核エキス及びその有効成分の影響を示す。 The results are shown in FIGS. 3 (a) to 3 (i). 3, active oxygen from neutrophils - shows the effect of loquat nuclear extract and its active ingredient on production (0 2).
対照例のラジカツト及びァスコルビン酸については、それぞれ図 4 (a)、 （b) に示す。  Figures 4 (a) and 4 (b) show the control of radicat and ascorbic acid, respectively.
これらの結果、 特に、 本発明の枇杷核エキス及び ;8—シトステロールは優れた 活性酸素産生抑制効果、 及び活性酸素除去効果を示した。 一方、 ラジカットは、 弱い活性酸素産生抑制効果を示したが、活性酸素除去効果は示さなかった。また、 ァスコルビン酸は、 活性酸素産生抑制効果を示さなかったが、 弱い活性酸素除去 効果を示した。  As a result, the loquat nucleus extract and; 8-sitosterol of the present invention showed excellent active oxygen production suppressing effect and active oxygen removing effect. On the other hand, Radicut showed a weak active oxygen production inhibitory effect, but did not show an active oxygen removing effect. Ascorbic acid did not show an active oxygen production inhibitory effect, but showed a weak active oxygen removing effect.
実施例 2 Example 2
次に、 活性酸素産生刺激剤として f ML Pの代わりに PMA (Phorbol myristate acetate (ホルポ一ルミリステート アセテート）） を用いて、 実 施例 1と同様に試験した。  Next, a test was conducted in the same manner as in Example 1, except that PMA (Phorbol myristate acetate) was used as a reactive oxygen production stimulating agent instead of fMLP.
結果を図 5 (a) 〜 （i) に示す。 対照例のラジカット及びァスコルビン酸に ついては、 それぞれ図 6 (a)、 （b) に示す。  The results are shown in Fig. 5 (a) to (i). Figures 6 (a) and 6 (b) show the results of Radicut and ascorbic acid in the control, respectively.
本発明の枇杷核エキスは活性酸素産生抑制効果及び活性酸素除去効果を示した。 β—シトステロールは弱い活性酸素除去効果が認められ、 活性酸素産生抑制効果 は認められなかった。 一方、 ラジカットはごく弱い活性酸素産生抑制効果及び活 性酸素除去効果を示したが、 ァスコルビン酸は効果が認められなかつた。  The loquat nucleus extract of the present invention exhibited an active oxygen production suppressing effect and an active oxygen removing effect. β-sitosterol showed a weak active oxygen removal effect, and no active oxygen production inhibitory effect. On the other hand, Radicut showed very weak active oxygen production inhibitory effect and active oxygen removal effect, but ascorbic acid had no effect.
実施例 3 Example 3
更に、 iMLPの代わりに AA (Arachidonic Acid (ァラキドン酸)） を用 いて、 実施例 1と同様に試験した。 In addition, use AA (Arachidonic Acid) for iMLP And tested in the same manner as in Example 1.
結果を図 7 ( a ) 〜（i )に示す。 対照例のラジカット及びァスコルビン酸につ いては、 それぞれ図 8 ( a )、 （b ) に示す。  The results are shown in Fig. 7 (a) to (i). Radicut and ascorbic acid in the control are shown in FIGS. 8 (a) and (b), respectively.
本発明の枇杷核エキスは、 濃度を高くした場合に活性酸素産生抑制効果を示し た。 また、 活性酸素除去効果を示した。 ]3—シトステロールは弱い活性酸素産生 抑制効果及び活性酸素除去効果を示した。 一方、 ラジカットは、 わずかに活性酸 素産生抑制効果を示し、 活性酸素除去効果は認められなかった。 また、 ァスコル ビン酸は効果が認められなかつた。  The loquat nucleus extract of the present invention exhibited an active oxygen production inhibitory effect when the concentration was increased. In addition, it showed an active oxygen removal effect. ] 3-Sitosterol showed weak active oxygen production inhibitory effect and active oxygen removal effect. Radicut, on the other hand, showed a slight inhibitory effect on active oxygen production and no active oxygen removal effect. Ascorbic acid had no effect.
実施例 4 Example 4
次に、 キサンチン ·キサンチンォキシダ一ゼによる活性酸素発生系について調 ベるために、 高水溶性テトラリゾゥム塩還元法を用いて枇杷核エキスの活性酸素 (02—)消去能について検討した。 Next, in order to investigate the active oxygen generation system using xanthine / xanthine oxidase, the active oxygen (0 2 —) scavenging ability of loquat nucleus extract was examined by using a highly water-soluble tetralithodium salt reduction method.
その結果を表 2に示す。 The results are shown in Table 2.
表 2 Table 2
枇杷核エキス、 その有効成分、 ラジカット及びァスコルビン酸 の SOD様活性SOD-like activity of loquat nucleus extract, its active ingredients, Radicut and ascorbic acid
^ァ 7χΛν SS〇Dυ/ ^ A 7χΛν SS〇Dυ /
Figure imgf000023_0001
Figure imgf000023_0001
リノレン酸 1 0 5 ± 2 2 市販医薬品一  Linolenic acid 1 0 5 ± 2 2
ァスコルビン酸 1 3 5±4 5 ラン力ッ卜 N S  Ascorbic acid 1 3 5 ± 4 5
(η = 6) 各枇杷核エキス、 有効成分及びラジカット、 ァスコルビン酸の濃度は 1 OOjt Mとした N S ： S O D活性は認、められなかった (η = 6) The concentration of each loquat core extract, active ingredient, Radicut, and ascorbic acid was 1 OOjt M. NS: No SOD activity was observed.
表 2から明らかなように、 本発明の枇杷核エキスは 10 O M濃度において、 SOD活性は 85%と高かったが、 有効成分、 ラジカット及びァスコルビン酸は 10%前後、 もしくは認められなかった。 As is clear from Table 2, the loquat nucleus extract of the present invention had a high SOD activity of 85% at a concentration of 10 OM, but the active ingredients, Radicut and ascorbic acid were around 10% or were not observed.
実施例 5 Example 5
また、安定ラジカル DPPH 1. l-Diphenyl-2-picrylhedrazyl (1. 1 -ジフエ二ルー 2—ピクリルヒドラジル)） を用いてフリーラジカル 「活性酸素 In addition, using the stable radical DPPH 1. l-Diphenyl-2-picrylhedrazyl (1.1-diphenyl-2-picrylhydrazyl))
(〇2_) +ハイドロキシラジカル （-〇H)」 の消去能について検討した。 50 DPPHのエタノール溶液に、 エタノール又は蒸留水に溶解した枇杷 核エキス及び有効成分を加え、室温下で 5分以内に吸光度 517 nmにて測定し、 枇杷核エキス及び各有効成分による DPPHの消去作用より、 50%(I C50) 消去量を求め、 ラジカットとァスコルピン酸との比較検討した。 The scavenging ability of (〇 2 _) + hydroxyl radical (-〇H) was examined. To a 50 DPPH ethanol solution, add loquat nucleus extract and active ingredient dissolved in ethanol or distilled water, measure at room temperature within 5 minutes at an absorbance of 517 nm, and remove the DPPH by loquat nucleus extract and each active ingredient. From this, the 50% (IC50) elimination amount was determined, and a comparison between Radicut and ascorbic acid was conducted.
その結果を表 3に示す。 The results are shown in Table 3.
表 3 Table 3
瞓^OSHdads /O。 瞓 ^ OSHdads / O.
Figure imgf000025_0001
表 3は、 枇杷核エキス及びその有効成分、 ラジカット、 ァスコルビン酸の DP PH50%消去量を示す。
Figure imgf000025_0001
Table 3 shows the loquat core extract, its active ingredient, Radicut, and the amount of DPPH 50% eliminated for ascorbic acid.
表 3から明らかなように、 本発明の枇杷核エキスは、 DPPH50%消去量が 1 4. 1 Mと低く、 対照としたラジカット及びァスコルビン酸も同様に 13. 7 Μ, 28.2 Μと低く認められた。 As is clear from Table 3, the loquat nucleus extract of the present invention has a DPPH 50% elimination amount of 1%. As low as 4.1 M, Radicut and ascorbic acid as controls were also found as low as 13.7Μ and 28.2Μ.
実施例 6 Example 6
次に、 ESR(Electron Spin Resonance (電子スピン共鳴)） を用いて枇杷 核エキス及び各有効成分のハイドロキシラジカル消去能について検討した。  Next, ESR (Electron Spin Resonance) was used to examine the ability of the loquat nucleus extract and each active ingredient to scavenge the hydroxyl radical.
0.1 15MPBN (phenyl t-b tyl nitrone) リン酸緩衝液に l OmM硫酸 アンモニゥム鉄 （I I) 水溶液を加えてハイドロキシラジカルを発生させ、 ES Rにて枇杷核エキス及び各有効成分のハイドロキシラジカル消去能を測定し、 ラ ジカツト及びァスコルビン酸との比較検討した。  0.1 15MPBN (phenyl tbtyl nitrone) Phosphate buffer is added with l OmM aqueous solution of ammonium iron (II) sulfate to generate hydroxyl radical, and ESR is used to measure loquat nucleus extract and the active radical scavenging ability of each active ingredient. And radicat and ascorbic acid.
その結果を表 4に示す。 The results are shown in Table 4.
表 4 Table 4
¾¾i^T。口」4^ν 枇杷核エキス、 その有効成分、 ジカツト及びァスコルビン酸 のヒドロキシラジカル消去活性 枇杷核エキス及び有効成分 消去活性 （％) 枇杷核エキス 32. 1 ± 3. 3 ¾¾i ^ T. Mouth ”4 ^ ν Loquat nucleus extract, its active ingredient, hydroxy radical scavenging activity of dicat and ascorbic acid Loquat nucleus extract and active ingredient scavenging activity (%) Loquat nucleus extract 32.1 ± 3.3
β—シトステロール 26. 5 ±4. 3  β-sitosterol 26.5 ± 4.3
β—シトス亍ロールモノゲリコシド 8. 6 ± 2. 6  β-cytosperol monogericoside 8.6 ± 2.6
アミグダりン 1 5. 0 ± 1. 0  Amygdin 15.0 ± 1.0
ベンズアルデヒド 1 5 3士 1. 5  Benzaldehyde 1 5 3rd person 1.5
マンデロニトリル 5. 2 ± "I . 1  Mandelonitrile 5.2 ± "I. 1
23. 5 ± 3. 4  23.5 ± 3.4
リノール酸 6. 5 ± 1. 3  Linoleic acid 6.5 ± 1.3
リノレン酸 1 2. 6 ± 1. フ 巿販医薬品  Linolenic acid 1 2. 6 ± 1.
ァスコルビン酸 26. 5土 1 8  Ascorbic acid 26.5 Sat 1 8
ラジカッ卜 22. 5 ± 1 3  Radicat 22.5 ± 1 3
(η = 6)  (η = 6)
Figure imgf000027_0001
Figure imgf000027_0001
表 4から明らかなように、 Fenton反応によるヒドロキシラジカルの消去活性 (ESR法） は、 枇杷核エキスが 32%、 β—シトステロールが 27%、 安息香 酸が 24%と高く、 対照としたラジカツト及びァスコルビン酸は 23 %及び 2 7 %とほぼ同様の消去活性を示した。 As is clear from Table 4, the hydroxyl radical scavenging activity (ESR method) by the Fenton reaction was as high as 32% for loquat nucleus extract, 27% for β-sitosterol, and 24% for benzoic acid. The acid showed almost the same erasing activity as 23% and 27%.
実施例 7 Example 7
次に 「肝機能検査値 （AST及び ALT) カ征常範囲を超えている被験者： 3 名」 を対象として本発明の健康飲食品の効果を調べた。  Next, the effect of the health food and drink of the present invention was examined on “3 subjects with liver function test values (AST and ALT) exceeding the control range.”
¾：杷核エキス未 10. 4gを、 400, 000m 1の水に溶かして調製し、 こ れを 1日 2回、 1回 100mlを 1週間、 被験者に食用させた。 結果を表 5に示 す。 表 5は、 肝機能検査値 （AST及び ALT) が正常範囲を超えている被験者 に対する効果を示す表である。  ¾: Hakone extract (10.4 g) was prepared by dissolving it in 400,000 ml of water, and this was eaten twice a day, 100 ml at a time, for one week for one week. Table 5 shows the results. Table 5 shows the effects on subjects whose liver function test values (AST and ALT) are outside the normal range.
表 5' Table 5 '
) n)/ πnisvνΊ, ) n) / πnisvνΊ,
CvJ LO  CvJ LO
CO LO O  CO LO O
寸 CD  Dimension CD
m卜  m
o o  o o
CO 寸 CM CO size CM
s: CO.  s: CO.
寸 寸 寸  Dimensions Dimensions
CVl CO CVl CO
その結果、 全ての被験者で改善が見られた。 As a result, improvement was observed in all subjects.
実施例 8 Example 8
次に、 「肝機能検査値（ァー GTP) が正常範囲を超えている被験者： 3名」 を 対象として本発明の健康飲食品の効果を調べた。 Next, “3 subjects with liver function test values (GTP) outside the normal range:” The effects of the health food and drink of the present invention were examined as subjects.
枇杷核エキス末 10. 4gを、 400， 000m 1の水に溶かして調製し、 こ れを 1日 2回、 1回 10 Omlを 3週間、 被験者に食用させた。 その結果を表 6 に示す。 表 6は、 肝機能検査値 （τ— GTP) が正常範囲を超えている被験者に 対する効果を示す表である。  Loquat nucleus extract powder (10.4 g) was prepared by dissolving it in 400,000 ml of water, and this was eaten twice a day, 10 Oml once a day for 3 weeks. Table 6 shows the results. Table 6 shows the effect on subjects whose liver function test value (τ-GTP) exceeds the normal range.
表 6 Table 6
Figure imgf000030_0001
その結果、 全ての被験者で改善が見られた。
Figure imgf000030_0001
As a result, improvement was observed in all subjects.
実施例 9 Example 9
「肝機能検査値が正常範囲内の被験者： 5 名」 を対象として本発明の健康飲食品 の効果を調べた。  The effect of the health food and drink of the present invention was examined for “5 subjects with liver function test values within the normal range”.
枇杷核エキス末 10. 44gを、 400， 00 Om 1の水に溶かして調製し、 こ れを 1日 2回、 1回 10 Omiを 1週間、 被験者に食用させた。 結果を表 7に示 す。 表 7は、 肝機能検査値が正常範囲内の被験者に対する効果を示す表である。 表 7
Figure imgf000031_0001
Loquat nucleus extract powder 10.44 g was prepared by dissolving in 400,00 Om 1 of water, and this was eaten twice daily and 10 Omi once a week for 1 week. Table 7 shows the results. Table 7 shows the effect on subjects whose liver function test values are within the normal range. Table 7
Figure imgf000031_0001
全ての被験者において正常範囲内であった。  It was within the normal range in all subjects.
実施例 1 0 Example 10
次に、 本発明の健康飲食品の効果をラットを用いて、 調べた ラッ卜の生化学検査値に及ぼす各種枇杷核エキスの影響 Next, the effects of the health food and drink of the present invention were examined using rats. Of various loquat nucleus extracts on biochemical test values of rat
雄性 Wistar系ラット、 7週齢を 1週間予備飼育した後、 健常ラットにっきジ メチル二トロサミン（40mg/kg)を単回腹腔内投与した。 これにより肝細胞 の線維化を誘発させた。  Male Wistar rats, 7 weeks of age, were preliminarily bred for 1 week, and healthy rats were given a single intraperitoneal injection of dimethylditrosamine (40 mg / kg). This induced hepatocyte fibrosis.
ジメチルニトロサミン投与後 7日の肝障害ラッ卜に対して、 調製した各種ェキ スを倍量の水に溶解し、 この水溶液を、 給水瓶にて自由摂取させた。 また、 正常 ラッ卜についても同様に投与した。  For the liver injury rats 7 days after dimethylnitrosamine administration, the various prepared excipients were dissolved in twice the amount of water, and this aqueous solution was freely taken in a water bottle. In addition, normal rats were similarly administered.
各水溶液のエキス含量は、 それぞれ、 70%エタノールエキスは 0. 5%、 メ 夕ノールエキスは 0. 45%、水ェキスは0. 75 %、へキサンエキスは 0. 2% であった。  The extract content of each aqueous solution was 0.5% for 70% ethanol extract, 0.45% for methanol extract, 0.75% for water extract, and 0.2% for hexane extract, respectively.
各種エキス投与後の正常ラット及び肝障害ラットについて、 投与後 7日にラッ トを屠殺し、 Vi S IONアナライザ一（ダイナポット社製）を用い、 血清中の glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) 値を測定した。  Rats were sacrificed 7 days after administration of normal rats and hepatic impaired rats after administration of various extracts. Serum glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate Transaminase (GPT) values were measured.
結果を図 9に示す。 普通資料投与群の G〇T、 GPT値は、 正常ラットに比べ 著しく高値を示した。 枇杷核エキス投与群の GOT値は、 溶媒により差が認めら れ、 特に、 70%エタノール及びメタノールエキス投与群の場合、 普通飼料投与 群に比べ低値を示すことが認められた。 水及びへキサンエキス投与群の場合にお いても、 普通飼料投与群と比較して効果が認められた。  Fig. 9 shows the results. The G〇T and GPT values in the group receiving the normal data were significantly higher than those in normal rats. The GOT value of the loquat nucleus extract-administered group showed a difference depending on the solvent. In particular, it was observed that the 70% ethanol and methanol extract-administered group showed lower values than the normal feed-administered group. The effect was also observed in the group to which water and hexane extract were administered compared to the group to which normal feed was administered.
したがって、 G〇T、 GPT値の値から、 肝障害の進行が抑えれていると考え られる。  Therefore, the G 考 え T and GPT values suggest that the progression of liver injury is suppressed.
実施例 11 Example 11
ラッ卜の肝レチノィド量に及ぼす各種枇杷核エキスの影響  Effect of Various Loquat Nucleus Extracts on Liver Retinoid Content in Rats
肝障害ラットの作成、 及び各種エキスの投与は、 実施例 10にしたがって行つ た。 各種エキス投与後の正常ラット及び肝障害ラットについて、 投与後 7日に肝 臓中のパルミチン酸レチノール量を測定した。 ラット摘出肝の一定部位を約 0. 3 g秤量し、 クロ口ホルム 5mlを加え、 氷冷下、 細胞粉碎機 (アイラード社製） を用いてホモジネート（10, 000回転、 2分間）を行ない、遠心分離（3， 50 0回転、 20分間)後、 下層を分取し、 試料とした。 これら試料を 0. 5 ΙΠメン ブランフィルター（ミリポア社製)で濾過し、 HP LC用試料とした。 Preparation of hepatopathy rats and administration of various extracts were performed according to Example 10. Seven days after administration, the amount of retinol palmitate in the liver was measured for normal rats and hepatic impaired rats after administration of various extracts. Approximately 0. 3 g is weighed, 5 ml of black-mouthed form is added, and the mixture is homogenized (10,000 rpm, 2 minutes) using a cell disrupter (manufactured by Eilard) under ice-cooling, and centrifuged (3,500 rpm, 20 rpm). After that, the lower layer was separated and used as a sample. These samples were filtered through a 0.5ΙΠ membrane filter (manufactured by Millipore) to obtain samples for HP LC.
測定条件は、 次の通りである。 日立 L 6000高速液体クロマトグラフ  The measurement conditions are as follows. Hitachi L 6000 High Performance Liquid Chromatograph
検出 日立 L 3000UV検出器 Detection Hitachi L 3000UV detector
検出波長 310 nm - カラム Cosnoseal 5C 18 (150x4.6匪 i. DNAcarai Tesque Co., Ltd. Detection wavelength 310 nm-Column Cosnoseal 5C 18 (150x4.6 Maraud i. DNAcarai Tesque Co., Ltd.
測定温度 Measurement temperature
移動相 テトラヒドロフラン：メタノール（1 ： 3) Mobile phase Tetrahydrofuran: methanol (1: 3)
流速 1. 0ml / i n 結果を図 10に示す。 肝レチノィド量は、 予備飼育を行った直後の正常ラット の肝レチノィド量を 100%とし、 それに対する割合で示した。 肝レチノィド量 は、 普通飼料投与群において、 正常ラットに比べ著しく低値を示した。 一方、 ェ キス投与群においては溶媒により、 肝レチノイド量に差が認められた。 特に、 7 0%エタノール、 メタノール及び水エキス投与群の場合、 普通飼料投与群に比べ 肝レチノイド量の増大が認められた。 へキサンエキス投与群では、 普通飼料投与 群との差は認められなかった。 Flow rate 1.0 ml / in Figure 10 shows the results. Liver retinoid content was expressed as a percentage of the liver retinoid content of normal rats immediately after pre-breeding, assuming 100%. Liver retinoid levels were significantly lower in the normal diet group than in normal rats. On the other hand, a difference was observed in the amount of hepatic retinoids in the group administered with the solvent depending on the solvent. Particularly, in the group to which 70% ethanol, methanol and water extract were administered, the amount of hepatic retinoid was increased as compared to the group to which normal feed was administered. No difference was observed in the hexane extract administration group from the normal feed administration group.
実施例 12 Example 12
ラットのハイドロキシプロリン量に及ぼす各種枇杷核エキスの影響  Effects of various loquat nucleus extracts on the amount of hydroxyproline in rats
肝障害ラットの作成、 及び各種エキスの投与は、 実施例 10にしたがって行つ た。 ラット摘出肝の一定部位を約 0. 3g枰量し、 エタノール 5m Iを加え、 氷冷 下、細胞粉砕器 (アイラード社製）を用いてホモジネ一ト（10， 000回転、 2分 間）を行い、 遠心分離（3， 500回転、 20分間)後、 上清を分取した。 この液 1 m 1を枰取し、 60°Cで 8時間加温乾固した。残渣に、エタノール 40 ^ 0. lmo 1ホウ酸緩衝液 80 l を加え溶解した後、 蛍光ラベル化剤である 100 mmo 1 4一フルオロー 7—二トロべンゾフラザン（NBD'F)を 40 1添カロ し、 室温遮光下、 15時間反応させ、 蛍光ラベル化処理を行なった。 この液に、 5m mo 1塩酸 840 1 を加え反応停止後、 遠心分離（3， 500回転、 20 分間）を行ない上清を 0. 5^mメンブランフィル夕一（ミリポア社製)で濾過し、 HP LC用試料とした。 Preparation of hepatopathy rats and administration of various extracts were performed according to Example 10. Approximately 0.3 g of a fixed part of the rat isolated liver was weighed, 5 ml of ethanol was added, and the mixture was homogenized (10,000 rpm, 2 minutes) using a cell crusher (manufactured by Eilard) under ice-cooling. After centrifugation (3,500 rpm, 20 minutes), the supernatant was collected. 1 ml of this solution was taken out and heated to 60 ° C. for 8 hours to dryness. The residue was dissolved by adding 80 l of ethanol 40 ^ 0. lmo 1 borate buffer, and then added with a fluorescent labeling agent, 100 mmo 14 monofluoro 7-2-nitrobenzofurazan (NBD'F). Then, the mixture was reacted at room temperature under light shielding for 15 hours to perform a fluorescent labeling treatment. To this solution was added 5 mM hydrochloric acid 840 1 to stop the reaction, followed by centrifugation (3,500 rpm, 20 minutes). A sample for HP LC was used.
測定条件は、 次の通りである。  The measurement conditions are as follows.
装置 ： 日立 L 6000高速液体クロマトグラフ Equipment: Hitachi L6000 High Performance Liquid Chromatograph
検出 ： 日立 L 7480蛍光光度計 Detection: Hitachi L 7480 Fluorometer
検出波長 ： Ex 475 nm Em 530 nm Detection wavelength: Ex 475 nm Em 530 nm
カラム ： YMC Pack ODS-AQ 150X6. Omm I.D. Column: YMC Pack ODS-AQ 150X6. Omm I.D.
測定温度 Measurement temperature
移動相 ァセトニトリル:水（35： 65→ 50 ： 50への 15分間のグ ラジュェン卜） Mobile phase acetonitrile: water (15 min gradient from 35:65 to 50:50)
1.0 m 1 / m i n 結果を図 11に示す。 ハイドロキシプロリン量は、 普通飼料投与群において、 正常ラットに比べ著しく高値を示した。 一方、 エキス投与群においては溶媒によ り、 肝レチノイド量に差が認められた。 特に、 70%エタノール、 メタノール及 び水エキス投与群の場合、 普通飼料投与群に比べハイドロキシプロリン量の減少 が認められた。 へキサンエキス投与群の場合、 普通飼料投与群に比べ多少の差が 認められた。 実施例 1 3 FIG. 11 shows the results of 1.0 m 1 / min. The amount of hydroxyproline was significantly higher in the normal diet group than in normal rats. On the other hand, in the extract-administered group, a difference was observed in the amount of hepatic retinoid depending on the solvent. In particular, in the group to which 70% ethanol, methanol and water extract were administered, the amount of hydroxyproline was reduced compared to the group to which normal feed was administered. In the hexane extract-administered group, a slight difference was observed compared to the normal feed-administered group. Example 13
障害ラットの肝線維化率に及ぼす各種枇杷核エキスの影響  Effect of various loquat nucleus extracts on the rate of liver fibrosis in impaired rats
肝障害ラットの作成、 及び各種エキスの投与は、 実施例 1 0にしたがって行つ た。  Preparation of hepatopathy rats and administration of various extracts were performed according to Example 10.
各種エキス投与ラットの肝線維化率は、 ラット摘出肝を Azan - Mal lory染色 法にて染色した肝組織像よりコンピュータ画像解析にて青色に染色した肝線維化 部位の面積を算出し、 対照群 (普通飼料投与ラット）との肝線維化面積との比によ り求めた。  The rate of liver fibrosis in rats treated with various extracts was calculated by calculating the area of liver fibrosis sites stained blue by computer image analysis from liver tissue images stained with Azan-Mallory staining of isolated rat liver. It was determined from the ratio of the liver fibrosis area to that of a normal feed-treated rat.
結果を図 1 2に示す。 肝線維化率は、 普通飼料投与群の線維化面積を 1 0 0 % として、 それに対する比で示した。 肝線維化率は、 枇杷核エキス投与群が、 普通 飼料投与群に比べ低値を示した。 また、 その値は、 溶媒により差が認められ、 7 0 %エタノール及びメタノールエキス投与群の場合、 水及びへキサンエキス投与 群より低値を示した。  The results are shown in FIG. The hepatic fibrosis rate was expressed as a ratio to the fibrosis area of the normal diet administration group, which was defined as 100%. The liver fibrosis rate was lower in the group receiving loquat extract than in the group receiving normal feed. Further, the value was different depending on the solvent, and the value was lower in the 70% ethanol and methanol extract administration groups than in the water and hexane extract administration groups.
肝障害ラットに、 枇杷核エキスを経口投与した場合、 普通飼料投与群に比べ、 生化学検査値 (G O T, G P T) (図 9参照)、 レチノイド量（図 1 0参照）、 ハイ ドロキシプロリン量 （図 1 1参照） 及び線維化率 （図 1 2参照） の有意な改善が 認められた。  When loquat nucleus extract was orally administered to rats with hepatic impairment, the biochemical test values (GOT, GPT) (see Fig. 9), the amount of retinoids (see Fig. 10), (See Fig. 11) and significant improvement in the fibrosis rate (see Fig. 12).
実施例 1 4 Example 14
正常家兎の血中脂質量の影響  Effect of blood lipid level in normal rabbits
正常家兎への投与は、日本白色種雄性家兎（ 1 . 8— 2 k g)を 1週間予備飼育し た後、 健常家兎につき 1群 6羽として、 普通飼料 ·飲料水投与群、 普通飼料 '枇 杷核エキス投与群の 2群に分け、 4週間投与を行った。  For normal rabbits, Japanese white male rabbits (1.8-2 kg) were preliminarily reared for 1 week, and then 6 rabbits per group per normal rabbit. The feed was divided into two groups, the loquat nucleus extract administration group, and administered for 4 weeks.
各家兎の耳静脈より、 投与開始後、 1週、 2週、 3週及び 4週にそれぞれ採血 を行い血清を採取した。 これらの血清について、 V I S I ONアナライザー （ダ イナポット（製)） を用い、 G O T値、 G P T値、 総コレステロール値、 L D Lコ レステロール値、 HD Lコレステロール、 中性脂肪値（トリグリセライド値)等の 生ィ匕学検査値を調べた。 各検査値は、 6羽の兎から得た値の平均値と標準誤差で 示し、各検査値の枇杷核エキス投与群と対象群との差について、 s tudentの t検 定により 5 %の危険率で有意差検定を行った。 結果を図 1 3に示す。 Blood was collected from the ear vein of each rabbit at 1 week, 2 weeks, 3 weeks and 4 weeks after administration, and serum was collected. For these sera, using VISI ON Analyzer (Dynapot (manufactured)), GOT, GPT, total cholesterol, LDL cholesterol, HD L cholesterol, triglyceride, etc. The values of the laboratory tests were examined. Each test value is shown as the average value and the standard error of the values obtained from 6 rabbits, and the difference between each test value between the loquat nucleus extract-administered group and the control group was 5% dangerous by the student t test. The significance test was performed by the rate. The results are shown in FIG.
図 1 3の結果から明らかなように、 正常家兎に枇杷核エキスを投与した場合、 枇杷核エキスを投与しない場合と比較して、 動脈硬化を促進させる要因となって いる血清総コレステロール値及び L D Lコレステロール値は、 低値を示した。 実施例 1 5  As is evident from the results in Fig. 13, when the loquat nucleus extract was administered to normal rabbits, the serum total cholesterol level and the factors that promoted atherosclerosis were higher than when the loquat nucleus extract was not administered. LDL cholesterol levels were low. Example 15
コレステロール含有飼料投与家兎の血中脂質量の影響  Effect of blood lipid level in rabbits fed cholesterol-containing diet
上述の手順に従い枇杷核エキスを調製した。 コレステロール含有飼料投与家兎 への投与は、 予備飼育後の家兎を 1群 6羽として、 1 %コレステロール含有飼料 (清水実験材料 (株)製） ·飲料水投与群及び 1 %コレステロール含有飼料 ·枇杷核 エキス投与群の 2群に分け、 4週間投与を行った。 なお、 1 %コレステロール含 有飼料の投与量は、 1日 7 5 g (コレステロールとして 3 7. 5〜4 1 . 7 mg/ k g)とした。  Loquat nucleus extract was prepared according to the above procedure. Cholesterol-containing feed-administered rabbits were preliminarily reared into 6 rabbits per group, and 1% cholesterol-containing feed (manufactured by Shimizu Experimental Materials Co., Ltd.) The loquat nucleus extract was divided into two groups and administered for 4 weeks. The dose of the diet containing 1% cholesterol was 75 g / day (37.5 to 41.7 mg / kg as cholesterol).
実施例 1 4と同様に、 生化学検査値を測定した。 結果を図 1 4に示す。  Biochemical test values were measured in the same manner as in Example 14. The results are shown in FIG.
図 1 4の結果から明らかなように、 コレステロール含有飼料投与家兎に枇杷核 エキスを投与した場合、 枇杷核エキスを投与しない場合と比較して、 動脈硬化を 促進させる要因となっている血清総コレステロール値及び L D Lコレステロ一ル 値は、 低値を示した。  As is clear from the results in Fig. 14, when loquat nucleus extract was administered to rabbits to which a cholesterol-containing feed was administered, serum total, which is a factor promoting arteriosclerosis, was higher than when loquat nucleus extract was not administered. Cholesterol and LDL cholesterol levels were low.
実施例 1 6 Example 16
高脂血症家兎の血中脂質量の影響  Effect of lipid content in blood of rabbits with hyperlipidemia
上述の手順に従い枇杷核エキスを調製した。まず、予備飼育後の家兎を用いて、 1 %コレステロール含有飼料 ·飲料水を 3週間投与して、 高脂血症家兎を作成し た。なお、 1 %コレステロール含有飼料の投与量は、 1日 7 5 g (コレステロール として 3 7 . 5〜4 1 . 7 m g Z k g)とした。 これらの家兎について、 投与飼料 を 1 %コレステロール含有飼料から普通飼料に変更した後、 1群 6羽として、 普 通飼料 ·飲料水投与、 普通飼料 ·枇杷核エキス投与群に分け、 4週間投与を行つ た。 Loquat nucleus extract was prepared according to the above procedure. First, pre-bred rabbits were treated with 1% cholesterol-containing feed and drinking water for 3 weeks to prepare hyperlipidemic rabbits. The dose of the 1% cholesterol-containing feed was 75 g per day (37.5 to 41.7 mg Z kg as cholesterol). For these rabbits, the feed was changed from 1% cholesterol-containing feed to normal feed, and then 6 rabbits per group were used. They were divided into groups for feed-feeding and drinking water administration, normal feed and loquat nucleus extract administration, and were administered for 4 weeks.
実施例 1 4と同様に、 生化学検査値を測定した。 結果を図 1 5に示す。  Biochemical test values were measured in the same manner as in Example 14. The results are shown in FIG.
図 1 5の結果から明らかなように、 高脂血症家兎に枇杷核エキスを投与した場 合、 枇杷核エキスを投与しない場合と比較して、 動脈硬化を促進させる要因とな つている血清総コレステロール値及び L D Lコレステロ一ル値は、低値を示した。 実施例 1 7  As is clear from the results in Fig. 15, the serum that is a factor that promotes atherosclerosis when loquat nucleus extract is administered to hyperlipidemic rabbits is greater than when loquat nucleus extract is not administered. Total cholesterol and LDL cholesterol levels were low. Example 17
高脂血症家兎の血中脂質量の影響  Effect of lipid content in blood of rabbits with hyperlipidemia
高脂血症家兎を、 1 %コレステロール含有飼料 ·飲料水を 4週間投与して作成 した以外、 実施例 1 6と同様に枇杷核エキスの投与を行った。  The loquat nucleus extract was administered in the same manner as in Example 16 except that hyperlipidemic rabbits were prepared by administering 1% cholesterol-containing feed and drinking water for 4 weeks.
実施例 1 4と同様に、 生化学的検査値を測定した。 結果を図 1 6に示す。 図 1 6の結果から明らかなように、 高脂血症家兎に枇杷核エキスを投与した場 合、 枇杷核エキスを投与しない場合と比較して、 動脈硬化を促進させる要因とな つている血清総コレステロール値及び L D Lコレステロール値は、低値を示した。  Biochemical test values were measured in the same manner as in Example 14. The results are shown in FIG. As is clear from the results in Fig. 16, the serum that is a factor that promotes atherosclerosis when loquat nucleus extract is administered to hyperlipidemic rabbits is greater than when loquat nucleus extract is not administered. Total and LDL cholesterol levels were low.
〔発明の効果〕 〔The invention's effect〕
本発明のフリーラジカル消去剤は、 有害なフリーラジカルを効率的に除去し得 るという有利な効果を有する。  The free radical scavenger of the present invention has an advantageous effect that harmful free radicals can be efficiently removed.
本発明のフリ一ラジカル消去剤は、 素材が天然素材であるため、 副作用を生じ ることなしに、 所望の効果を達成することができ、 医療への責献はもとより、 農 産物の利用範囲の拡大、 及び廃棄物の有効利用等、 社会的貢献度が高いという有 利な効果を奏する。  Since the free radical scavenger of the present invention is a natural material, it can achieve desired effects without causing side effects, and can not only contribute to medical treatment but also reduce the range of use of agricultural products. It has the beneficial effects of high social contribution, such as expansion and effective use of waste.
本発明の健康食品又は健康飲料は、 自然素材を単独で利用した場合では得られ ない、 健康維持、 促進するし得るという有利な効果を有する。 健康を維持、 促進 する効果として具体的には、 肝機能障害の予防 ·改善作用、 コレステロール溶解 作用、 高脂血症改善作用、 鎮咳去痰作用等の効果を、 副作用なしに得ることがで きるという有利な効果を奏する。 The health food or health drink of the present invention has an advantageous effect of being able to maintain and promote health, which cannot be obtained when natural materials are used alone. As an effect to maintain and promote health, specifically, it is possible to obtain effects such as prevention and improvement of liver dysfunction, cholesterol dissolving, hyperlipidemia improving, antitussive and expectorant effects without side effects. It has the advantageous effect of turning off.
本発明の健康食品又は健康飲料は、 日常的に摂取する食品又は飲料に添加する ことにより、 日常の食生活の中で肝炎、 肝硬変及ぴ抗脂血症等の疾患を予防する ことが可能であり、 また改善も期待できるという有利な効果を奏する。  The health food or health drink of the present invention can prevent diseases such as hepatitis, cirrhosis and hypolipidemia in a daily diet by being added to foods or drinks to be taken daily. There is an advantageous effect that improvement can be expected.

Claims

請 求 の 範 囲 The scope of the claims
1 . 枇杷核由来エキスを有効成分とするフリーラジカル消去剤。 1. Free radical scavenger containing loquat nucleus extract as an active ingredient.
2 . 枇杷核由来エキスが、枇杷核を粉碎して得た粉砕物を、 エタノール、 メ夕ノ ール、 τΚ、 へキサン、 又はこれらの混合物からなる群から選択される少なくとも 1種の溶媒に浸漬して、 上清を分取して得られることを特徴とする請求項 1記載 のフリーラジカル消去剤。  2. The loquat nucleus-derived extract is used to pulverize the loquat nucleus into a crushed product, which is then mixed with at least one solvent selected from the group consisting of ethanol, methanol, τΚ, hexane, and mixtures thereof. The free radical scavenger according to claim 1, wherein the free radical scavenger is obtained by immersing and collecting the supernatant.
3 . 枇杷核由来エキスが、 リノール酸、 リノレン酸、 /3—シトステロール、 β— シトステロール—3— 0—モノグリコシド、 アミグダリン、 ベンズアルデヒド、 マンデニトリル、 安息香酸からなる群から選択される少なくとも 1種を含有する ことを特徴とする請求項 1又は 2項に記載のフリーラジカル消去剤。  3. The loquat nucleus-derived extract comprises at least one selected from the group consisting of linoleic acid, linolenic acid, / 3-sitosterol, β-sitosterol-3-0-monoglycoside, amygdalin, benzaldehyde, mandenitrile, and benzoic acid. The free radical scavenger according to claim 1, wherein the free radical scavenger is contained.
4 . 枇杷核由来エキスが、 i3—シトステロール及び |3—シトステロ一ルー 3— 0 一モノグリコシドを含有することを特徴とする請求項 1〜 3項のいずれか 1項に 記載のフリーラジカル消去剤。  4. The free-radical scavenger according to any one of claims 1 to 3, wherein the loquat nucleus-derived extract contains i3-sitosterol and | 3-sitosterol mono-3-o-glycoside. .
5 . 枇杷核由来エキスを含有する健康飲食品。  5. Healthy food and drink containing loquat nucleus-derived extract.
6 . ftl:杷核由来エキスが、枇杷核を粉碎して得た粉砕物を、 エタノール、 メ夕ノ —ル、 水、 へキサン、 又はこれらの混合物からなる群から選択される少なくとも 1 種の溶媒に浸漬して、 上清を分取して得られることを特徴とする請求項 5記載 の健康飲食品。  6. ftl: Haku nucleus extract extracts ground material obtained by pulverizing loquat nucleus into at least one kind selected from the group consisting of ethanol, methanol, water, hexane, or a mixture thereof. The health food or drink according to claim 5, which is obtained by immersing in a solvent and collecting a supernatant.
7 . 枇杷核由来エキスが、 リノール酸、 リノレン酸、 ）3—シトステロール、 β— シトステロール— 3— 0—モノグリコシド、 アミグダリン、 ベンズアルデヒド、 マンデニトリル、 安息香酸からなる群からなる少なくとも 1種を含有することを 特徴とする請求項 5又は 6項に記載の健康飲食品。  7. The loquat nucleus-derived extract contains at least one selected from the group consisting of linoleic acid, linolenic acid,) 3-sitosterol, β-sitosterol-3-0-monoglycoside, amygdalin, benzaldehyde, mandenitrile, and benzoic acid. The health food or drink according to claim 5 or 6, wherein
8 · 枇杷核由来エキスが、 β—シトステロ一ル及び] 3—シトステロール一 3— 0 一モノダリコシドを含有することを特徴とする請求項 5〜 7項のいずれか 1項に 記載の健康飲食品。 8. The health food or drink according to any one of claims 5 to 7, wherein the loquat nucleus-derived extract contains β-sitosterol and] 3-sitosterol- 13-0-monodaricoside.
9 . 健康飲食品が、健康食品である請求項 5〜8項のいずれか 1項に紀載の健康 飲食品。 9. The health food or drink according to any one of claims 5 to 8, wherein the health food or drink is a health food.
1 0 . さらに、 米粉、 そば粉、 うどん粉、 小麦粉、 パン粉からなる群から選択さ れる少なくとも 1種を含有する請求項 9項に記載の健康飲食品。  10. The health food or drink according to claim 9, further comprising at least one selected from the group consisting of rice flour, buckwheat flour, udon flour, and bread crumbs.
1 1 . 健康飲食品が、健康飲料である請求項 5〜 8項のいずれか 1項に記載の健 康飲食品。  11. The health food or drink according to any one of claims 5 to 8, wherein the health food or drink is a health drink.
1 2 . さらに、 水、 ココア、 コーヒー、 紅茶、 牛乳、 酒、 果実飲料、 又は野菜 飲料を含有する請求項 1 1記載の健康飲食品。  12. The health food or drink according to claim 11, further comprising water, cocoa, coffee, tea, milk, alcohol, fruit drink, or vegetable drink.
PCT/JP2003/001951 2002-02-22 2003-02-21 Free radical quenching agent contaiing eriobotryae seed extract as the active ingredient and health foods and drinks containing eryobotryae seed extract WO2003070022A1 (en)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103584112A (en) * 2013-10-18 2014-02-19 宿州市皖神面制品有限公司 Production method of Chinese wolfberry health-care vital gluten
US11432552B2 (en) * 2017-03-07 2022-09-06 Rilis Co., Ltd. Deodorizing/antibacterial agent

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH06237736A (en) * 1993-02-16 1994-08-30 Kozaburo Shiba Functional fruit composition of rosaceous plant
JPH10295325A (en) * 1997-04-28 1998-11-10 Katsuji Nagamitsu Health food

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH06237736A (en) * 1993-02-16 1994-08-30 Kozaburo Shiba Functional fruit composition of rosaceous plant
JPH10295325A (en) * 1997-04-28 1998-11-10 Katsuji Nagamitsu Health food

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103584112A (en) * 2013-10-18 2014-02-19 宿州市皖神面制品有限公司 Production method of Chinese wolfberry health-care vital gluten
US11432552B2 (en) * 2017-03-07 2022-09-06 Rilis Co., Ltd. Deodorizing/antibacterial agent

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