WO2007142096A1

WO2007142096A1 - Inhibitor of inducible nitric oxide synthase, beverage/food, and neutraceutical food

Info

Publication number: WO2007142096A1
Application number: PCT/JP2007/060984
Authority: WO
Inventors: Shigeaki Ohno; Kazuhiro Ogami; Takako Nogami; Mari Kamegai; Yoshikiyo Ono; Akiko Yokota; Kazunaga Yazawa
Original assignee: Organo Corporation; Eye Ambitious, Co., Ltd.
Priority date: 2006-06-02
Filing date: 2007-05-30
Publication date: 2007-12-13

Abstract

Disclosed is an inducible nitric oxide synthase inhibitor which has few adverse side-effects and can be taken easily. Also disclosed is a beverage/food having an inhibitory effect on an inducible nitric oxide synthase. The inhibitor comprises a Bacopa monniera extract as an active ingredient. The inhibitor may comprises a polyphenol from an olive fruit as an active ingredient.

Description

Specification

Inhibitors of inducible nitric oxide synthase, food and drink, and nutritional supplements

[0001] The present invention relates to an inhibitor of inducible nitric acid-nitrogen synthase containing bacopa monniera extract as an active ingredient, a food and drink and a dietary supplement that exhibits an inhibitory function of inducible nitric oxide synthase.

[0002] The present invention relates to an inhibitor of inducible nitric oxide synthase comprising olive fruit polyphenol as an active ingredient, a food and drink and a nutritional supplement that exhibits an inhibitory function of inducible nitric oxide synthase.

Background art

[0003] Nitric acid-nitrogen has a wide range of effects on living tissues. While it plays a role in biological defense such as blood pressure regulation and information transmission control, excessive synthesis can cause inflammation and carcinogenesis. Nitric oxide is synthesized by a plurality of nitric oxide synthases in living tissues, and in particular, it is known that inducible nitric acid nitric acid synthase synthesizes a large amount of nitric acid nitric acid. The

[0004] On the other hand, if a chemical substance that can control the oversynthesis of nitric oxide is searched, there is a high possibility that carcinogenesis and inflammation can be suppressed. In particular, natural ingredients that are applicable to foods have a mild effect and can be taken by a wide range of patients. In addition, even for healthy people, the possibility of preventing many diseases such as diabetes and arteriosclerosis derived from carcinogenesis and inflammatory power is increased by daily intake.

[0005] Incidentally, there has been disclosed for the jig療方method utilizing pharmaceutical and inhibitory action have an inhibitory effect of nitric oxide synthase (e.g., see Patent Document 1, 2.) ₀ Also, natural a material, some of which have been disclosed to have an inhibitory action of Ichisani匕窒containing synthase (e.g., see Patent Document 3-6.) ₀

[0006] Bacopa monniera is a herb that grows naturally in the lowland wetlands of India, and has been regularly used in India as a remedy for aurveda. Contains many types of flavonoids and saponins

Especially, it has been said to be highly effective in brain activation and rejuvenation. In the test using rats Has been demonstrated to increase the anti-acid activity. In addition, cholinesterase inhibitory activity was also observed in mice. In administration trials for patients with anxiety, the effects of reducing anxiety, regaining confidence, and improving concentration are prominent in the treatment group. In addition, there are reports that improvement in learning ability was observed when taken to school-aged children. In addition, it has been reported that insomnia and irritability are improved, memory is maintained, and the prognosis of brain diseases is effective.

[0007] Olive is a plant native to the Mediterranean region, and its fruit is rich in eating experience. It contains many kinds of polyphenols and has strong anti-acid activity, so it has been said to maintain and promote health since ancient times. In particular, in research on polyphenols in fruit juice, a wide range of antioxidant activity has been observed in all tissues of the body, including the activity of suppressing the oxidation of low-density lipoprotein in the blood. The fact that people in the Mediterranean region prefer meat and greasy meals but low vascular disease such as arteriosclerosis has led to frequent consumption of polyphenols in olive juice. In addition, it has been reported that it is effective in preventing diabetes, cancer, cranial nerve diseases such as Alzheimer's and Parkinson's disease, and skin roughness.

[0008] Patent Document 1: JP 2003-137776

Patent Document 2: Special Table 2005-506986

Patent Document 3: JP 2004-161664

Patent Document 4: JP-A-2005-213242

Patent Document 5: JP 2006-83149 A

Patent Document 6: JP-A-2005-35981

Disclosure of the invention

Problems to be solved by the invention

[0009] However, it is not known that bacopa monniera extract (an extract of bacopamo-ela) or olive fruit polyphenol has an inhibitory effect on inducible nitric acid-nitrogen synthase

[0010] The present invention relates to the side effects of bismuth subgallate described in Patent Document 1, such as when ingested by a person with constipation, a patient who has undergone a colon fistula or a patient with a digestive tract diverticulum. Inducible monoacid-nitrogen synthase inhibitor and inducible type 1 that are mild in action and less likely to cause vomiting, loss of appetite, gingivitis, and even severe cases. The object is to provide a food or drink having a function of inhibiting nitric oxide synthase.

Means for solving the problem

[0011] As a result of diligent studies to achieve the above-mentioned object, the present inventors have found that Bacopa monniera extract is a fruit-free polyphenol with an inhibitory action of inducible nitric oxide synthase.

V. Discovered previously unknown attributes, and found that bacopa monniera extract and olive fruit polyphenols are suitable for use in new applications (for example, prevention and control of inflammation and carcinogenesis). Based on this, the present invention has been completed.

[0012] An inhibitor of inducible nitric oxide synthase as an exemplary aspect of the present invention is characterized in that Bacopa moniera extract is an active ingredient.

[0013] Another exemplary aspect of the present invention is an inhibitor of inducible monoacid-nitrogen synthase comprising an olive fruit polyphenol as an active ingredient.

[0014] In still another exemplary aspect of the present invention, an inhibitor of inducible nitric acid-nitrogen synthase comprises Nocopa monniera extract and olive fruit polyphenol as active ingredients. This inhibitor of inducible monoxide-nitrogen synthase may be one obtained by further adding at least one of copper and zinc.

[0015] Copper and zinc are added, and the weight ratio of the added copper and zinc is 1: 1 or more 1

: Desirable to be 200 or less 1: 4 or more 1: Desirable if 100 or less 1: 1: 4 or more

1: 50 or less is more desirable.

[0016] A food and drink as another exemplary aspect of the present invention is characterized by adding a Bacopa monniera extract exhibiting an inhibitory function of inducible monoacid-nitrogen synthase.

[0017] The weight ratio of Nocopa monniera extract is preferably 20% or less based on the total weight of the food and drink.

[0018] A food and drink as still another exemplary aspect of the present invention is characterized by the addition of olive fruit polyphenol which exhibits an inhibitory function of inducible monoacid-nitrogen synthase.

[0019] The weight ratio of olive fruit polyphenol is 20% or less based on the total weight of the food and drink. More desirable! /

[0020] Still another exemplary aspect of the present invention is a food or drink product comprising a bacopa monniera extract and olive fruit polyphenol that exhibit an inhibitory function of inducible monoacid-nitrogen synthase. It is characterized by.

[0021] The total weight of Nocopa monniera extract and olive fruit polyphenol is more preferably 40% or less with respect to the total weight.

[0022] The food or drink may be obtained by further adding at least one of copper and zinc.

[0023] It is desirable to add copper and zinc, and the weight ratio of the added copper and zinc should be 1: 1 or more and 1: 200 or less. If it is 1: 4 or more and 1: 100 or less More desirable 1: 4 or more and 1:50 or less are more desirable.

[0024] A dietary supplement as still another exemplary aspect of the present invention is characterized by comprising a Bacopa monniera extract that exhibits an inhibitory function of inducible monoacid-nitrogen synthase.

[0025] It is desirable that the weight ratio of Nocopa monniera extract is 5% or more and 98% or less with respect to the total weight.

[0026] A dietary supplement as still another exemplary aspect of the present invention is characterized by adding olive fruit polyphenol that exhibits an inhibitory function of inducible monoacid-nitrogen synthase.

[0027] The weight ratio of olive fruit polyphenol is desirably 5% or more and 98% or less based on the total weight.

[0028] A dietary supplement as still another exemplary aspect of the present invention comprises the addition of Nocopa monniera extract and olive fruit polyphenol that exerts the inhibitory function of inducible monoacid-nitrogen synthase. It is characterized by becoming.

[0029] The total weight of Nocopa monniera extract and olive fruit polyphenol is preferably 5% or more and 98% or less based on the total weight.

[0030] This dietary supplement may be obtained by further adding at least one of copper and zinc. [0031] It is desirable that copper and zinc are added, and the weight ratio of the added copper and zinc is 1: 1 or more and 1: 200 or less. More desirable 1: 4 or more and 1:50 or less are more desirable.

[0032] Copper is a component of various enzymes and acts as a catalyst for oxygen transport, electron transfer, redox, and the like. Insufficient copper is said to cause anemia, leukopenia and bone abnormalities, and excessive intake may cause liver damage. According to the Japanese dietary intake standard in 2005, the recommended amount is 0.3 to 0.8 (additional amount of pregnant women and lactating women 0.1, 0.6) mg / day (depending on age and sex), upper limit is lOmgZ days And the copper in the composition should not exceed lOmg / day! /, Which is preferred U.

[0033] Zinc is also a component of various enzymes and is involved in biological reactions such as gene expression and protein synthesis. Zinc deficiency is said to result in loss of appetite and taste disorders, and overdose may cause gastric disorders, dizziness, nausea in acute poisoning, anemia in the chronic, immune disorders, neurological symptoms, diarrhea, etc. is there. According to the Japanese dietary intake standard in 2005, the recommended amount is 2 to 10 (pregnant and lactating supplements 3) mgZ days (depending on age and sex), and the upper limit is 30 mg / day. Should not exceed 30 mg / day.

[0034] By taking zinc and copper at the same time, it is expected to increase the absorption of active ingredients such as bacopa monniera extract and olive fruit polyphenol into the body. In order to prevent copper absorption, the weight ratio of copper to zinc is preferably in the range of 1: 1 to 1: 200, more preferably 1: 4 to 1: 100, more preferably 1 : 4 or more and 1:50 or less.

[0035] In addition, if necessary, vitamin C, vitamin E, vitamin A, which has anti-acidic action, rutin, zeaxanthin, α-lipoic acid, ant, considered to be beneficial for maintaining eye health You may add cyanine, anthocyanin, wastaxanthin, yew leaf extract, 13 carotene, lycopene, or omega-3 fatty acids such as EPA and DHA.

[0036] Nutritional supplements here are all foods that are useful for maintaining and improving health. Health functional foods (specific health foods, nutritional functional foods), health supplements, nutrition-enriched foods, nutritional adjustments Including food and supplements. Additives such as excipients, binders, disintegrants, solubilizers In addition, it can be made into dosage forms such as tablets, capsules, pills, powders, granules, syrups, and injections by known production techniques.

[0037] The weight ratio of Nocopa monniera extract is preferably 2% or more and 98% or less based on the total weight, although it depends on the dosage form. In particular, when it is used as a tablet or capsule, it is preferably 5% or more and 80% or less.

[0038] The weight ratio of olive fruit polyphenol is preferably 2% or more and 98% or less based on the total weight, although it depends on the dosage form. In particular, when it is used as a tablet or capsule, it is preferably 5% or more and 75% or less.

The invention's effect

[0039] An inhibitor of inducible mono-acid-nitrogen synthase and bacopa monniera ex- hibite the inhibitory function of inducible mono-acid-nitrogen synthase of the present invention comprising bacopa monniera extract as an active ingredient The foods and drinks that are added include Altono with less toxicity and side effects, prevention of Imah's disease and Parkinson's disease, reduction of symptoms, improvement of mental disorders such as anxiety, atopic dermatitis, sunburn, burns, etc. Skin diseases, allergic keratoconjunctivitis, iritis, ciliitis, scleritis, inflammatory eye diseases such as photopathy, glomerulonephritis, pyelonephritis, prostatitis, bronchitis, pneumonia, gastroenteritis, rheumatoid arthritis It is useful for prevention and symptom relief of autoimmune diseases such as osteoporosis, colon cancer, Behcet's disease and collagen disease.

[0040] An inducible nitric oxide synthase inhibitor containing olive fruit polyphenol of the present invention as an active ingredient and an olive fruit polyphenol exhibiting an inhibitory function of inducible nitric oxide synthase are added. Foods and drinks are vascular diseases such as arteriosclerosis and hypertension with minimal toxicity and side effects, skin diseases such as atopic dermatitis, sunburn, burns, allergic keratoconjunctivitis, iritis, ciliitis, scleritis Inflammatory eye diseases such as age-related macular degeneration, diabetic retinopathy, cataracts caused by light damage, glomerulonephritis, pyelonephritis, prostatitis, tracheositis, pneumonia, gastroenteritis, rheumatoid arthritis, osteoporosis, colon cancer, It is useful for preventing diseases caused by ocular circulation such as Behcet's disease and collagen diseases such as autoimmune diseases, regulatory eye strain, regulatory convulsions and glaucoma, and diabetic macular edema.

[0041] In addition, since it is possible to prevent various inflammations and cancers by simply taking foods and beverages to which Bacopa monniera extract oriolberry fruit polyphenol is added, it is not necessary to take preventive drugs. Even simple. It can be easily ingested daily as a food and drink, and it is easy to make it a habit. You need to be careful about your intake.

[0042] Furthermore, Bacopa monniera extract olive fruit polyphenol has a bitter taste, and when added to food or drink, it has the effect of deepening the taste of the food or drink and increasing the sense of quality. The bacopa monniera extract should be a food or drink for inhibiting inducible nitric acid nitric acid synthase that has an appropriate bitter taste without impairing the flavor of the food or drink by keeping the amount of added force below a certain level. Can do.

BEST MODE FOR CARRYING OUT THE INVENTION

[0043] The Bacopa monniera extract, which is an active ingredient of the present invention, can be obtained by drying the whole plant of Bacopamo-ela and extracting the extract with ethanol. Specifically, dry whole grass is pulverized and extracted with ethanol at about 60 ° C to 70 ° C. Filter the extract to remove solids and dry to make Bacopa monniera extract. As a drying method, evaporation, spray drying, freeze drying, or the like can be used. The Bacopa monniera extract used here was extracted at 65 ° C for 15 minutes with 3 times the volume of ethanol in the ground whole dried Bacopamo-ela. After extraction, the ethanol was changed and extracted again with new 3 volumes of ethanol at 65 ° C for 15 minutes for a total of 3 extractions. A total of 9 times the total amount of the extract, which had been filtered to remove solids, was combined and concentrated on a rotary evaporator, and finally dried in an oven at 65 ° C for testing.

[0044] The olive fruit polyphenol, which is an active ingredient of the present invention, is obtained by squeezing olive fruit with water to which a reducing agent is added in an amount equal to the weight of the fruit, and removing the oil content by squeezing the pulp. The obtained aqueous layer can be obtained by drying by a method such as evaporation, spray drying or freeze drying. The purpose of adding a reducing agent to the water in the fruit is to prevent the browning of the olive fruit polyphenol due to the acidity of the olive fruit polyphenol. Any reducing agent can be used as long as it fulfills its purpose, and vitamin C 'cysteine and citrate are suitable. Here, the olive fruit polyphenol preparation actually used for the test was added with water with a citrate concentration of 1% added to the olive fruit, squeezed, separated the aqueous layer, and then the aqueous layer was separated. It was obtained by freeze-drying. This lyophilized olive fruit polyphenol can be used for food and drink applications. As another drying method, a method such as spray drying can also be used.

[0045] Inhibitors of inducible nitric acid-nitrogen synthase containing Bacopa monniera extract olive oil polyphenol as an active ingredient according to the present invention have an anti-inflammatory action, for example, its immunomodulatory action, etc. It is useful as an immunomodulator. Applicable diseases that can be prevented or alleviated include allergic keratoconjunctivitis, iritis, ciliary inflammation, scleritis, keratitis due to light disorder, cataract, retinal diseases such as age-related macular degeneration, inflammatory eye Diseases, atopic dermatitis, hay fever, glomerulonephritis, pyelonephritis, prostatitis, bronchitis, pneumonia, gastroenteritis, rheumatoid arthritis, osteoporosis, colon cancer, Behcet's disease, collagen disease, etc. Examples include ophthalmic regulatory convulsions due to cancer and fatigue. In addition, the inhibitor of inducible nitric oxide synthase containing bacopamo-ela extract according to the present invention as an active ingredient is applicable to prevention of eye strain or alleviation of symptoms.

[0046] Inhibitors of inducible nitric oxide synthase containing Bacopa monniera extract olive fruit polyphenol as an active ingredient according to the present invention include additives such as excipients, binders, disintegrants, and solubilizers. In addition, tablets, capsules, powders, granules, syrups by known formulation techniques

Or a dosage form such as an injection.

[0047] In addition, the food and drink according to the present invention is manufactured by adding Bacopa monniera extract or olive fruit polyphenol which exhibits an inhibitory function of inducible nitric oxide synthase to various foods and drinks. Can do. For example, examples of food and drink include soft drinks, confectionery, frozen confectionery, dairy products, alcoholic beverages, and meats. Bacopamoniella extract and olive fruit polyphenol have some bitterness, so they are inherently bitter (such as chiyocholate and cocoa), tasteful food (such as western liquor), and stimulating. Foods and beverages (eg carbonated drinks) are more desirable because they can relatively reduce the change in taste caused by the addition of Nocopa monniera extract or olive fruit polyphenol.

[0048] The appropriate intake of Bacopa monniera extract is 50 to 50 OOmgZ per adult for oral intake, preferably 200 to 1000mgZ days of effective ingestion range Various types of disease prevention and symptom relief Therefore, the dose can be adjusted according to the condition of the person taking the food.

[0049] The appropriate intake of olive fruit polyphenol is 5 per adult for oral intake. A 0~5000mgZ曰, monkey Preferably, the force ^s is effective uptake range 100~1000mgZ曰, depending on the state of those who ingested for the prevention or relief of symptoms of various diseases, in increased or decreased appropriately child transgression .

[0050] Further, the content of Bacopa monniera extract olive fruit polyphenol as an effective component of the inhibitory function of inducible nitric oxide synthase in the food and drink according to the present invention is as follows. The content can be specified so that the normal intake satisfies the above-mentioned appropriate intake or effective intake. On the other hand, as the amount added to food and drink increases, the bitterness becomes stronger and there is a higher possibility of adversely affecting the original taste of food and drink.Therefore, the amount of bacopa monniera echis and olive fruit polyphenol is added to the total weight of the food and drink. It is desirable to keep the weight ratio to 20% or less.

[0051] Hereinafter, the present invention will be described by way of examples, but the present invention is not limited thereto.

Example

[0052] [Example 1]

<Confirmation of inhibitory action of Bacopa monniera extract on inducible monoxide-nitrogen synthase> As a test group, mouse macrophage-derived cells, RAW264. 7 cells, 5 x 10 ⁴ cells per well plate In contrast, a 1% dimethyl sulfoxide (DMSO) -containing culture solution adjusted so that the final concentration of Bacopa moniera extract is 1 _μg / ml, 10 g / ml, 100 ^ gZml, Each was added. The RAW264.7 cells after the addition of this culture solution are pre- 24 hours in the presence of 37 ° C and 5% CO.

2

Cultured. The DMSO-containing culture solution is a mixture of 1/10 volume of immobilized fetal bovine serum in RPMI-1640 medium supplemented with 2 mM glutamine, 100 U / ml penicillin, and 1 OO ml streptomycin.

[0053] After washing RAW264. 7 cells with phosphate buffered saline (PBS), 10 g of lipopolysaccharide (LPS: Salmonella typhimurium Sigma— Aldrich per 1 ml culture in the presence of the same concentration of Bacopa monniera extract ) Was added. Cells (macrophages) were stimulated by culturing for 24 hours.

[0054] Thereafter, expression of inducible nitric oxide synthase was examined by Western plotting. It was. The group stimulated with LPS but not added bacopa monniera extract (hereinafter referred to as P group) and the control group not stimulated with LPS and also added no bacopa monniera extract (hereinafter referred to as Q group! / ヽ) .) Was also provided. In either case, DMSO was added to a final concentration of 0.01%. After 24 hours of stimulation, TNF-alpha release into the medium and expression of inducible nitric oxide synthase were examined. The release of TNF-alpha is observed to increase especially when cells are stimulated. The TNF-alpha concentration was analyzed by ELISA, and the expression level of inducible monoacid-nitrogen synthase was analyzed by Western plotting.

[0055] FIG. 1 shows the detection result of TNF-alpha concentration. Compared to the Q group as a control group, the TNF-alpha concentration was significantly increased in the P group, and TNF-alpha was observed to be released by stimulation of cells by LPS. On the other hand, in the Bacopa monniera ex added group (hereinafter referred to as “A group”), the TNF-alpha concentration decreased with the increase in the amount of bacopa monniera supplement, and the bacopa monniera extract increased in a concentration-dependent manner. —It was confirmed that the release of alpha into the medium was suppressed. In group A, the amount of added Bacopa monniera exk 1 μgZml is group A1, the amount of bacopamoniella extract added 10 μgZ ml is group A2, and the amount of bacopa monniera extract added The group of 100 μgZml was designated as A3 group.

[0056] FIG. 2 shows expression level marks 21-25 indicating the expression level of inducible nitric oxide synthase in each test group (Group A, Group P, Group Q). FIG. 2 shows that the smaller the expression level mark, the more the inducible monoacid-nitrogen synthase is inhibited and the expression level becomes smaller. The expression level mark 21 is the Q group as the control group, the expression level mark 22 is the P group, the expression level mark 23 is the A1 group as the bacopa monniera extract addition group, and the expression level mark 24 is the bacopa monniera extract addition group The A2 group and the expression level mark 25 correspond to the A3 group as the Bacopa monniera extract added group.

[0057] The expression level mark 25 corresponding to the A3 group is smaller than the expression level mark 22 corresponding to the P group, and the expression of inducible nitric acid-nitrogen synthase is suppressed in the Nocopa monniera extract. Function was recognized. From the above results, it was confirmed that the release of TNF-alpha was inhibited by the Nocopa monniera extract and the expression of inducible nitric acid-nitrogen synthase was inhibited.

[Example 2] <Confirmation of anti-inflammatory effect of bacopamoniera extract on rat endotoxin (LPS) -induced uveitis>

Uveitis was induced by administering 200 g of LPS dissolved in 200 μl of sterilized distilled water subcutaneously in both hind footpads of 6-week old Lewis male rats (body weight 200-250 g). Thereafter, bacopa monniera exi was dissolved in 2 ml of PBS (containing 0.1% dimethyl sulfoxide) per kg of body weight of the rat and administered to the rat. The dose of Nocopa monniera extract was adjusted to 1 mg, 10 mg, and lOO mg, respectively, per kg body weight of the rat, and the tail vein was administered immediately after LPS administration, and this was administered (hereinafter referred to as B group). ). In addition, among the B groups, the group administered with lmg bacopamoniella extract per kg body weight of rat B1 group, the group administered with 1 Omg bacopamoniella extract per kg body weight of rat B group, the rat body weight lkg Group B3 was the group that received lOOmg of Nocopamonella extract. The control group (hereinafter referred to as R group) without LPS administration and the non-administration group (hereinafter referred to as S group) not administered with Bacopa monniera extract had 2 ml PBS (0.1% dimethyl) per kg body weight of rats. (Containing sulfoxide) was administered on the same time schedule as in Group B. The number of cases was 8 per group. When uveitis is induced, inflammatory cells such as macrophages infiltrate into the aqueous humor and infiltrate (increased calories), and the protein concentration increases. Here, rat endotoxin (LPS) is synonymous with lipopolysaccharide (LPS).

[0059] <Inflammation inhibitory effect of Bacopa monniera extract>

Anterior aqueous humor was collected 24 hours after LPS administration, and the number of inflammatory cells was counted with a hemocytometer. Figure 3 shows the measurement results.

[0060] When uveitis was induced by administration of LPS, U and inflammatory cells increased significantly in the anterior aqueous humor (see group S in FIG. 3). Inflammatory cell counts force O. 0 ± 0 per Maebosui lml the R group as the control group. 0 X to 10 was a ^five, increasing the 9 ± 17. 9 X 10 ⁵ cells 50. In S group I ran out. On the other hand, in the administration group (Group B) to which Bacopa monniera extract was administered, the number of infiltrating inflammatory cells was suppressed, particularly in the B2 and B3 groups, corresponding to the increase in the dose. In other words, the number of inflammatory cells per ml of anterior aqueous humor is 48.1 ± 11.9 X 10 ^{5 in} group B1, 3 2. 7 ± 2. 1 X 10 ^{5 in} group B2, and 3.6 in group B3. ± 0. 4 X 10 ⁵ pieces. [0061] Furthermore, the protein concentration was measured using a simple protein measurement kit manufactured by Pierce. Figure 4 shows the measurement results.

[0062] The protein concentration in the anterior aqueous humor increased by about 70-fold to 20.3 ± 3.6 mg in the S group compared to 0.3 ± 0.lmg per ml in the R group as a control group. did. On the other hand, in the administration group (Group B), an inhibitory effect on the protein concentration in the anterior aqueous humor was observed, particularly in the B2 and B3 groups, in order to correspond to the increased dose. In other words, the B2 group had a protein concentration of 15.7 ± 3.4 mg per ml of anterior aqueous humor, and the B3 group had a protein concentration of 8.2 ± 1.4 mg per ml of anterior aqueous humor.

[0063] The results of Example 1 show that Bacopa monniera extract suppresses the expression of inducible nitric oxide synthase induced by LPS stimulation at the cellular level and is specifically observed in stimulated cells. Has been shown to inhibit the release of TNF-alpha. In addition, the results of Example 2 show that the uveitis of rats induced by stimulation with Nocopa monniera extract LPS is markedly suppressed. Therefore, it was possible to propose a mechanism that Bacopa monniera extract suppresses the overexpression of inducible nitric oxide synthase and further suppresses the inflammatory response.

[0064] [Example 3]

A healthy adult volunteer was administered 300 mg of Bacopa monniera extract per day, and a questionnaire regarding the constitution and physical condition and a questionnaire regarding mental condition and eye strain were conducted before the start of administration and after one month of administration. A control group to which no Bacopa monniera extract was administered was also provided, and a questionnaire was conducted according to the same time schedule as the administration group. There were 55 patients in the treatment group and 23 in the control group.

[0065] The questionnaire regarding the constitution and physical condition is as follows. Easy to peel off, easy to remove hair, no hair stiffness, no lustrous hair, easy constipation, easy to diarrhea, easy to swell, cold, anemia, easy to dizzy or dizzy, tired Easy, poor sleep, light sleep, poor appetite, easy to get a fever, easy to catch a cold, low physical fitness , Low energy, easy to get frustrated, fun no matter what, motivated

29 questions for “V”.

[0066] Of the 29 items provided in the questionnaire regarding constitution and physical condition, the effect of improving sleep was particularly high in the administration group. In other words, 63% and 48% of the respondents answered that they had improved on the two items “bad sleep” and “slow sleep”. As for other items, the percentage of those who answered that it was improved was uniformly less than 15%. In the control group, the percentage of respondents who reported improvement was less than 15% for all items.

[0067] The questionnaire on mental state and eye strain is: “Eye is tired!”, Eyes hurt, Eyes are blurred, Tears are easy to tear, Eyes are red, Things are flickering, Shoulders and hips Irritated, easily irritated, prone to heavy head, prone to headache, poor vision, narrow vision, feeling heavy eyelids, painful back of eyes, prone to hyperemia Drowsiness, dry eyes, blurred focus, waking up, yesterday's weather, unable to remember yesterday's dinner, no person's name, no concentration, reading books, irritability “I feel vague anxiety driven by“

[0068] Of the 25 items provided in the mental and eye strain questionnaire, improvement in eye strain and irritability was significantly observed in the administration group. In other words, 30% of the respondents answered that their eyes were tired, and 34% of the respondents answered that they were irritated. For other items, the percentage of respondents who reported improvement was less than 10%. In the control group, the percentage of respondents who reported improvement was less than 15% for all items.

[0069] Administration studies on humans confirmed that Bacopa monniera extract acts in the brain to promote mental stability and sleep. Moreover, although it is suggested that eye strain is reduced, it is considered that this is based on the inflammatory reaction inhibitory effect confirmed in Example 1 and Example 2.

[0070] [Example 4]

Hereinafter, bacopamo that exhibits the inhibitory function of the inducible monoacid-nitrogen synthase according to the present invention. A method for producing a marble cookie as an example of a food or drink obtained by adding Niera extract will be described. This marble cookie originally uses cocoa powder, which has a bitter taste, and even if the cocoa dough is made by adding bacopa monniera extract, the change in the taste of the cookie can be kept relatively low. If possible, it is convenient because it gives a deeper and more luxurious taste to the cookies.

[0071] <Ingredients and weight ratio of marble cookies supplemented with Bacopa monniera extract> [0072] [Table 1]

Raw material and its weight ratio in plain fabric

1. Mix the note well until it becomes whitish cream (both plain and cocoa dough)

2. While mixing, add sugar 2-3 times to each, and when smooth, salt, egg yolk Add each.

3. For plain dough, add 1Z3 amount of flour and add it to the butter. If you are a cocoa farmer, mix the flour and cocoa powder, then add 1Z3 amount to the batch.

4. Add the remaining soft flour or a mixture of soft flour and cocoa powder while sieving and mix gently until the powder is gone.

5. Take out the dough from each ball and cool it in the refrigerator for about 40 minutes.

6. Tear 2 kinds of chilled dough into 3-4 pieces and combine them together (plain dough: cocoa dough = 1: 1).

7. Make the dough into a cylindrical shape with a diameter of 3-4cm and put it in a freezer and harden it.

8. When the dough is cool and hard, cut it into 7mm width.

9. Place in an oven at 180 ° C and bake for 10-15 minutes.

[0075] In Example 4, when the weight ratio of the bacopamoniella extract to the cocoa dough exceeds 0.6%, the bitterness of the bacopamoniella extract becomes too strong, and the flavor of the marble cookie becomes too strong. Damaged.

[Example 5]

Hereinafter, a method for producing a raw chocolate as another example of a food or drink obtained by adding a Bacopa moniera extract exhibiting the inhibitory function of the inducible monoacid-nitrogen synthase according to the present invention will be described. This raw chocolate is originally made from chocolate with a bitter taste, cocoa powder, and rum that has a strong taste, and even if bacopa monniera extract is added, the change in taste is kept relatively low. If you can, it is convenient because it gives you a deep and high-class feel to the flavor of fresh chocolate.

[0077] <Raw Chocolate Raw Material with Bacopamoniera Extract and Weight Ratio> [0078] [Table 3] Raw chocolate raw materials and their weight ratio

[0079] <Method for producing raw chocolate supplemented with Bacopa monniera extract>

1. Engrave plate chocolate.

2. Warm the cream and chop the chopped chocolate to make it creamy.

3. Add bacopa monniera extract and rum and cool at 5 ° C for 30 minutes.

4. Divide the chilled portion into small pieces.

5. Sprinkle with cocoa powder.

[0080] In Example 5, when the weight ratio of the Bacopa monniera extract exceeds 0.6%, which is 0.6%, the bitter taste of the Bacopa monniera extract becomes too strong and the flavor of the raw chocolate is impaired.

[0081] [Example 6]

<Confirmation of the inhibitory action of olivine nitrogen synthase on olive fruit polyphenol

>

As a test group, RAW264. 7 cells, which are derived from mouse macrophages, were adjusted so that the number of cells was 5 x 10 ⁴ per well of a 24 well plate, while the final concentration of olive fruit polyphenol was l ^ u Culture solutions containing 1% dimethyl sulfoxide (DMSO) adjusted to gZml, lO ^ g / ml, and 100 gZml were added, respectively. Then, the RAW264.7 cells after addition of this culture medium are pre-treated in the presence of 37 ° C and 5% CO for 24 hours.

2

Incubate for hours. The DMSO-containing culture solution is a mixture of 1/10 volume of immobilized fetal bovine serum in RPMI-1640 medium supplemented with 2 mM glutamine, 100 U / ml besylin, and 100 ml streptomycin. .

[0082] After washing RAW264. 7 cells with phosphate buffered saline (PBS), 10 µg of lipopolysaccharide (LPS: per ml of culture medium in the presence of olive fruit polyphenol at the same concentration) Salmonella typhimurium Sigma—Aldrich) was added. Cells (macrophages) were stimulated by culturing for 24 hours.

[0083] Subsequently, expression of inducible nitric oxide synthase was examined by Western plotting. There is also a group stimulated with LPS but not added olive fruit polyphenol (hereinafter referred to as T group) and a control group (hereinafter referred to as U group) which is not stimulated with LPS nor added with olive fruit polyphenol. Provided. In either case, DMSO was added to a final concentration of 0.01%. After 24 hours of stimulation, TNF-alpha release into the medium and expression of inducible nitric oxide synthase were examined. TNF-alpha release is observed to increase specifically when cells are stimulated. The TNF-alpha concentration was determined by ELISA, and the expression level of inducible nitric oxide synthase was determined by Western plotting.

FIG. 5 shows the detection result of the TNF-alpha concentration. Compared with the U group as a control group, the TNF-alpha concentration was significantly increased in the T group, and TNF-alpha was observed to be released by stimulation of cells by LPS. On the other hand, in the olive fruit polyphenol addition group (hereinafter referred to as group C), the TNF-alpha concentration decreased with the increase in the olive fruit polyphenol addition, and the olive fruit polyphenola concentration dependent TNF— It was confirmed that the release of alpha into the medium was suppressed. Of the group C, the group with olive fruit polyphenol added 1 gZml, group C1, the group with olive fruit polyphenol added 10 μg Zml, group C2, and the addition of olive fruit polyphenol. The group with an amount of 100 μg Zml was designated as C3 group.

[0085] FIG. 6 shows expression level marks 31 to 35 indicating the expression level of inducible nitric oxide synthase in each test group (group C, group T, group U). FIG. 6 shows that the smaller the expression level mark, the more the inducible monoxide-nitrogen synthase is inhibited and the lower the expression level. The expression mark 31 is the U group as a control group, the expression mark 32 is the T group, the expression mark 33 is the C1 group as the olive fruit polyphenol addition group, and the expression mark 34 is the olive fruit polyphenol addition. The C2 group as a group and the expression level mark 35 correspond to the C3 group as a group added with olive fruit polyphenol.

[0086] Compared with the expression level mark 32 corresponding to the T group, the expression level corresponding to the C1, C2, and C3 groups No. 33, 34, and 35 were smaller, and olive fruit polyphenols were found to have a function to suppress the expression of inducible nitrogen monoxide synthase. From the above results, it was confirmed that olive fruit polyphenol suppresses the release of TNF-alpha and inhibits the expression of inducible nitric acid-nitrogen synthase.

[0087] [Example 7]

Uveitis was induced by administering 200 g of LPS dissolved in 200 μl of sterilized distilled water subcutaneously in both hind footpads of 6-week old Lewis male rats (body weight 200-250 g). Thereafter, olive fruit polyphenol was dissolved in 2 ml of PBS (containing 0.1% dimethyl sulfoxide) per kg of body weight of the rat and administered to the rat. Olive fruit polyphenols were adjusted to 1 mg, lOmg, and lOOmg, respectively, per lkg body weight of the rat. It was. Of group D, group D1 was administered lmg olive fruit polyphenol per kg body weight of rat D, group D2 was administered lOmg olive fruit polyphenol per kg body weight of rat, group L2 was rat body weight lkg The group to which lOOmg of olive fruit polyphenol was administered was designated as D3 group. The control group (hereinafter referred to as V group) not administered with L PS and the non-administered group (hereinafter referred to as W group) not administered with olive fruit polyphenol were treated with 2 ml PBS (0.1% dimethyl) per kg of body weight of rats. Sulphoxide-containing) was administered on the same time schedule as group D. The number of cases was 8 per group. When uveitis is induced, inflammatory cells such as macrophages and T lymphocytes infiltrate (increased calories) in the aqueous humor and increase the protein concentration. Here, rat endotoxin (LPS) is synonymous with lipopolysaccharide (LPS).

[0088] <Inflammatory inhibitory effect of olive fruit polyphenol>

Anterior aqueous humor was collected 24 hours after LPS administration, and the number of inflammatory cells was counted with a hemocytometer. Figure 7 shows the measurement results.

[0089] When uveitis was induced by administration of LPS, a marked increase in U and inflammatory cells was observed in the anterior aqueous humor (see group W in Fig. 7). Inflammatory cells per ml of anterior aqueous humor in group V as a control group The power was SO. 0 ± 0. 0 X 10 ⁵ even, while the W group increased to 50.9 ± 15. 9 X 10 ⁵ even. On the other hand, in the administration group (Group D) to which olive fruit polyphenol was administered, the number of infiltrating inflammatory cells was suppressed to correspond to the increased dose. That is, the number of inflamed cysts per ml of anterior aqueous humor in the D1 group is ί or 42.1 ± 13. 9 X 10 ⁵ ; in the D2 group is 2.5 or 2 ± 2.1 X 10 ⁵ ; in the D3 group is 0. It was 5 ± 0.4 × 10 ⁵ pieces.

[0090] Furthermore, the protein concentration was measured using a simple protein measurement kit manufactured by Pierce. Figure 8 shows the measurement results.

[0091] The protein concentration in the anterior aqueous humor increased by about 70-fold to 20.3 ± 3.6 mg in the W group compared to 0.3 ± 0.lmg per ml in the V group as a control group. did. On the other hand, in the administration group (Group D), an inhibitory effect on the protein concentration in the anterior aqueous humor was observed, particularly in the D2 and D3 groups, in order to correspond to the increased dose. That is, in the D2 group, the protein concentration was 10.6 ± 3.4 mg per ml of anterior aqueous humor, and in the D3 group, the protein concentration was 5.8 ± 1.4 mg per ml of anterior aqueous humor.

[0092] The results of Example 6 suppressed the expression of inducible nitric oxide synthase induced by LPS stimulation at the olive fruit polyphenola cell level and were specifically observed in the stimulated cells. It has been shown to inhibit the release of TNF-alpha. In addition, the results of Example 7 show that rat uveitis induced by stimulation of olive fruit polyphenola LPS is markedly suppressed. Therefore, we could propose a mechanism that olive fruit polyphenol suppresses overexpression of inducible nitric oxide synthase and further suppresses the inflammatory response.

[0093] [Example 8]

The effect of olive fruit polyphenol on blood vessel age was analyzed in human volunteers. Olive fruit polyphenols processed into tablets were administered to adult volunteers at a dose of 300 mg per person per day. Blood vessel age was measured before the start of administration and one month after the start of administration. On the other hand, a control group to which no olive fruit polyphenol was administered was also provided, and the blood vessel age was measured according to the same schedule as the administration group. 54 patients in the treatment group, 26 in the control group It was a name.

As a measuring device, an acceleration pulse wave meter (BLOOD CIRCULATION CHECKER: manufactured by FUTU RE WAVE) was used. This device measures the state of peripheral blood circulation. The measurement results are from the younger vascular age: A +, A, A-, B +, B + X, B, BX, C +, C, B 1, B— X, C 1, C——, E + , E, D +, D, D—, E—, F, F—, G, G—. Each evaluation was statistically processed with a score from 20 to 0 from A to GZG-. Scoring power after 1 month of olive fruit polyphenols It was.

[0095] As shown in Fig. 9 (a) and (b), in the administration group administered olive fruit polyphenol, the proportion of those with rejuvenated vascular age was 64.8%. In the control group that did not, the proportion of those with young blood vessels was only 50%.

[0096] Administration studies on humans have confirmed that olive fruit polyphenols can improve vascular age and are useful for the prevention and symptom relief of vascular diseases such as arteriosclerosis and hypertension.

[0097] [Example 9]

Using UV Illuminator TM20 manufactured by Funakoshi Co., Ltd. (planar light source equipped with 6 UV lamps with a wavelength of 302 nm, voltage of 100 V, and output of 8 W), UV irradiation is applied to the subject's arm to improve pigmentation due to sunburn. The effect was confirmed.

[0098] A black rubber sheet (not shown) having 12 holes continuously covered one arm of the subject and irradiated with UV for 3 minutes from a distance of 26 cm from the arm (control test). Prior to irradiation, all holes were concealed with adhesive tape to prevent UV irradiation of the skin under the holes. The adhesive tape with a single hole was removed at 15-second intervals to expose the skin, and the UV irradiation time was increased according to the order of the holes. The UV irradiation site of this arm corresponding to each hole position was also set as irradiation site lp to irradiation site 12p in order of the short direction of irradiation time.

[0099] Seven hours after irradiation, the irradiated sites lp to 12p were observed to confirm inflammation due to sunburn. In this control test, of the 12 irradiation sites, the nine irradiation sites with the longer irradiation time, that is, irradiation Inflammatory reactions were observed at the 4p-12p shot sites (see Figure 10 (a)). After standing for 1 month and then observing pigmentation due to sunburn in the irradiated area, pigmentation was observed in all of the skins in the 4 irradiated to 12p irradiated areas (Fig. 10 (b) )reference). During the past month, the subject has not been administered olive fruit polyphenol.

[0100] Thereafter, the other arm of the same subject was irradiated with UV under the same conditions (effect confirmation test) to confirm the inflammatory reaction. Here, the UV irradiation part of the other arm corresponding to each hole position was also set as irradiation part lq to irradiation part 12q in order of the shortest irradiation time. As in the case of the control test, the force with a long irradiation time is 9 places, that is, the irradiated part 4c! Inflammatory reaction was observed at ~ 12q (see Fig. 10 (c)). In the effect confirmation test, after confirming the inflammatory reaction, subjects were administered 300 mg olive fruit polyphenol per day for 1 month, and after 1 month, the pigmentation at the irradiated site was examined. In the effect confirmation test with the administration of olive fruit polyphenol, pigmentation was caused in 7 places with a long irradiation time, that is, the irradiation site 6c! It was only about 12q of skin, and the amount of pigment deposited on the whole was remarkably suppressed (see FIG. 10 (d)).

[0101] [Example 10]

Hereinafter, a method for producing a marble cookie as an example of a food or drink obtained by adding olive fruit polyphenol exhibiting the inhibitory function of the inducible monoacid-nitrogen synthase according to the present invention will be described. This marble cookie originally uses cocoa powder with a bitter taste, and even if olive fruit polyphenol is added to make cocoa dough, the change in cookie taste can be kept relatively low. It is convenient because it gives a deeper and more luxurious taste to the cookies.

[0102] <Ingredients and weight ratio of marble cookies with olive fruit polyphenol added>

[0103] [Table 4] Raw material and its weight ratio in plain fabric

<Manufacturing method of marble cookie with olive fruit polyphenol> Mix well with butter until it becomes whitish cream (both plain and cocoa dough)

2. While mixing, add sugar 2 to 3 times, and when smooth, add salt and egg yolk.

3. For plain dough, add 1Z3 amount of flour and add it to the butter. For cocoa greens, mix the flour and cocoa powder in advance and add 1/3 of the powder to the butter.

5. Take out the dough from each ball and cool it in the refrigerator for about 40 minutes. 6. Tear 2 kinds of chilled dough into 3-4 pieces and combine them together (plain dough: cocoa dough = 1: 1).

8. When the dough is cool and hard, cut it into 7mm width.

9. Place in an oven at 180 ° C and bake for 10-15 minutes.

[0106] In Example 10, the weight ratio of olive fruit polyphenol to cocoa dough is 0.35%. When the strength exceeds 20%, the bitter taste of olive fruit polyphenol becomes too strong and the flavor of marble cookies is impaired. It is.

[Example 10]

Hereinafter, a method for producing raw chocolate as another example of a food or drink obtained by adding olive fruit polyphenol which exhibits the inhibitory function of inducible nitric oxide synthase according to the present invention will be described. This raw chocolate originally uses chocolate chocolate with a bitter taste, cocoa powder with a strong taste, and a strong taste of rum. If you can, it is convenient because it gives you a deeper and more luxurious feel to the taste of raw chocolate.

<0108] <Raw chocolate raw materials added with olive fruit polyphenol and its weight ratio

>

[0109] [Table 6]

Raw chocolate raw material and its weight ratio

1. Engrave plate chocolate.

2. Warm the cream and chop the chopped chocolate to make it creamy.

3. Add olive fruit polyphenol and rum and cool at 5 ° C for 30 minutes. 4. Divide the chilled portion into small pieces.

5. Sprinkle with cocoa powder.

[0111] In Example 11, the weight ratio of olive fruit polyphenol is 0.35%. When the force exceeds 20%, the bitter taste of olive fruit polyphenol becomes too strong and the flavor of raw chocolate is impaired. .

[0112] [Example 11]

A test drug was prepared by suspending Bacopa monniera extract, olive fruit polyphenol, copper dalconate, and zinc dalconate in 0.5 carboxymethylcellulose (CMC).

[0113] Oral administration was performed at 5 mlZkg for 24 hours, 3 hours, LPS administration, 1 hour, and 3 hours before LPS administration. The LPS group (Y group) and the control group (X group) received 0.5% CMC at the same time schedule.

[0114] Anterior aqueous humor was collected 24 hours after LPS administration, and the protein concentration and the number of cells were measured.

[0115] Table 7 shows the experimental conditions. The experimental results are shown in FIGS.

[0116] [Table 7]

From FIG. 11 and FIG. 12, when Nocopa monniera extract was used as the active ingredient, the effect of suppressing the protein concentration in the anterior aqueous humor was recognized by adding copper and zinc.

[0118] From Fig. 13 and Fig. 14, when olive fruit polyphenol is used as an active ingredient, the addition of copper and zinc suppresses the protein concentration in the aqueous humor and suppresses the number of inflammatory cells. It turns out that there is an effect.

From FIG. 15 and FIG. 16, when the Nocopa monniera extract and olive fruit polyphenol were used in combination, the effect of suppressing the number of inflammatory cells was observed.

Industrial applicability

[0120] According to the present invention, it is possible to provide humans and animals with pharmaceuticals and foods and drinks for the purpose of inhibiting an inducible nitric oxide synthase containing Bacopa monniera extract as an active ingredient.

[0121] According to the present invention, it is possible to provide humans and animals with pharmaceuticals and foods and drinks for the purpose of inhibiting an inducible nitric oxide synthase containing olive fruit polyphenol as an active ingredient. If people take these medicines and foods, they can relieve eye irritation, improve peripheral blood vessels, and suppress pigmentation after sunburn.

Brief Description of Drawings

[0122] [FIG. 1] A diagram illustrating the results of Example 1 of the present invention, which shows release of TNF-alpha from LPS-stimulated cells in RAW264. 7 cells, which are cells derived from mouse macrophages. It is a figure showing that Bacopa monniera extract suppresses.

FIG. 2 is a diagram for explaining the results of Example 1 of the present invention, in which derivates of inducible nitrogen monoxide synthase in cells stimulated with LPS in RAW264. 7 cells, which are cells derived from mouse macrophages. It is a figure showing that Bacopa monniera extract suppresses expression.

FIG. 3 is a diagram for explaining the results of Example 2 of the present invention, in which 6-week-old Lewis male rats (body weight 200-250 g) were dissolved in 200 μl of sterile distilled water subcutaneously in both hind footpads. It is a figure showing that Bacopa monniera extract suppresses the infiltration of the inflammatory cell into anterior aqueous humor in the uveitis induced by administering 200 μg of LPS.

FIG. 4 is a diagram for explaining the results of Example 2 of the present invention, in which 6-week-old Lewis male rats (body weight 200-250 g) were dissolved in 200 μ1 of sterile distilled water under the skin of both hind limbs It is a figure showing that Bacopa monniera extract suppresses the increase in the protein concentration in anterior aqueous humor in the uveitis induced by administering 200 μg of LPS.

FIG. 5 is a diagram for explaining the results of Example 6 of the present invention, in which TNF-alpha from cells stimulated with LPS in RAW264.7 cells, which are cells derived from mouse macrophages. It is a figure showing that an olive fruit polyphenol suppresses discharge | release of this.

FIG. 6 is a diagram illustrating the results of Example 6 of the present invention, in which derivates of inducible nitrogen monoxide synthase in cells stimulated with LPS in RAW 264.7 cells, which are cells derived from mouse macrophages. It is a figure showing that an olive fruit polyphenol suppresses an expression.

FIG. 7 is a diagram for explaining the results of Example 7 of the present invention, which was dissolved in 200 μ1 of sterile distilled water subcutaneously on both hind footpads of 6-week-old Lewis male rats (body weight 200-250 g). It is a figure showing that olive fruit polyphenol suppresses infiltration of inflammatory cells into anterior chamber water in uveitis induced by administration of 200 μg of LPS.

[8] FIG. 8 is a diagram for explaining the results of Example 7 of the present invention, in which 6-week-old Lewis male rats (body weight 200-250 g) were dissolved in 200 μl of sterilized distilled water subcutaneously in both hind footpads. It is a figure showing that olive fruit polyphenol suppresses the increase in the protein concentration in anterior aqueous humor in the uveitis induced by administering 200 μg of LPS.

9) A diagram illustrating the results of Example 8 of the present invention, in which a plurality of administration groups administered with olive fruit polyphenol for one month were compared with a plurality of control groups not administered with olive fruit polyphenol. In comparison, it shows that a marked improvement in vascular age was observed.

FIG. 10 is a diagram for explaining the results of Example 9 of the present invention, and shows how pigmentation due to sunburn is significantly reduced when olive fruit polyphenol is administered for 1 month. .

FIG. 11 is a diagram for explaining the results of Example 11 of the present invention, showing changes in protein concentration in anterior aqueous humor when bacopa monniera extract, copper and zinc are orally administered.

FIG. 12 is a diagram for explaining the results of Example 11 of the present invention, and showing changes in the number of inflammatory cells in anterior aqueous humor when bacopa monniera extract, copper and zinc are orally administered. FIG. 13 is a diagram for explaining the results of Example 11 of the present invention, and shows changes in protein concentration in anterior aqueous humor when orive fruit polyphenol, copper and zinc are orally administered.

14] A diagram for explaining the results of Example 11 of the present invention, in which olive fruit polyphenol It is a figure which shows the change of the number of inflammatory cells in an anterior aqueous humor at the time of administering orally, ru, copper and zinc.

FIG. 15 is a diagram for explaining the results of Example 11 of the present invention, in which the ratio of copper to zinc was 1:20, and the ratio of Nocopa monniera extract to olive fruit polyphenol was varied. It is a figure which shows the change of the protein concentration in an aqueous humor at the time of administering.

FIG. 16 is a diagram for explaining the results of Example 11 of the present invention, in which the ratio of copper to zinc was 1:20, and the ratio of Nocopa monniera extract to olive fruit polyphenol was changed to It is a figure which shows the change of the inflammatory cell number in the anterior aqueous humor at the time of administering.

Explanation of symbols

21-25: expression level mark

31-35: expression level mark

lp to 12p: Irradiation site

lq ~ 12q: Irradiation site

Claims

The scope of the claims

[I] An inhibitor of inducible nitric acid-nitrogen synthase containing Bacopa monniera extract as an active ingredient.

[2] An inducible monoacid-nitrogen synthase inhibitor comprising olive fruit polyphenol as an active ingredient.

[3] An inducible nitric oxide synthase inhibitor comprising Nocopa monniera extract and olive fruit polyphenol as active ingredients.

[4] The inducible monoxide-nitrogen synthase according to any one of claims 1 to 3, further comprising at least one of copper and zinc. Inhibitor.

[5] The inductive monoacid according to claim 4, wherein copper and zinc are added, and the weight ratio of the added copper and zinc is 1: 1 or more and 1: 200 or less.匕 Nitrogen synthase inhibitor.

6. The inductive monoacid according to claim 4, wherein copper and zinc are added, and the weight ratio of the added copper and zinc is 1: 4 or more and 1: 100 or less.匕 Nitrogen synthase inhibitor.

[7] The inductive monoacid according to claim 4, wherein copper and zinc are added, and the weight ratio of the added copper and zinc is 1: 4 or more and 1:50 or less. Inhibitor of nitrous acid synthase.

[8] A food and drink comprising an added bacopa monniera extract that exhibits an inhibitory function of inducible monoxide and nitrogen synthase.

[9] The food / beverage product according to claim 8, wherein the weight ratio of the Bacopa monniera extract is 20% or less based on the total weight.

[10] A food and drink prepared by adding olive fruit polyphenol which exhibits an inhibitory function of inducible monoacid-nitrogen synthase.

[II] The food or drink according to claim 10, wherein the weight ratio of the olive fruit polyphenol is 20% or less based on the total weight.

[12] A food and drink comprising a Nocopa monniera extract and olive fruit polyphenol that exerts an inhibitory function of inducible nitric oxide synthase.

[13] The food or drink according to claim 12, wherein the total weight of the Bacopa monniera extract and olive fruit polyphenol is 40% or less based on the total weight.

[14] The food or drink according to any one of claims 8 to 13, further comprising at least one of copper and zinc.

[15] Copper and zinc are added, and the weight ratio of added copper and zinc is 1: 1 or more 1:

The food or drink according to claim 14, wherein the food or drink is 200 or less.

[16] Copper and zinc are added, and the weight ratio of added copper to zinc is 1: 4 or more 1:

The food or drink according to claim 14, wherein the food or drink is 100 or less.

[17] Copper and zinc are added, and the weight ratio of added copper to zinc is 1: 4 or more 1:

The food or drink according to claim 14, wherein the food or drink is 50 or less.

[18] A dietary supplement comprising a Bacopa monniera extract that exhibits an inhibitory function of inducible monoxide-nitrogen synthase.

[19] The dietary supplement according to claim 18, wherein the weight ratio of the Bacopa monniera extract is 5% or more and 98% or less based on the total weight.

[20] A dietary supplement supplemented with olive fruit polyphenol that exhibits the inhibitory function of inducible monoacid-nitrogen synthase.

21. The dietary supplement according to claim 20, wherein a weight ratio of the olive fruit polyphenol is 5% or more and 98% or less with respect to the total weight.

[22] A dietary supplement supplemented with Nocopa monniera extract and olive fruit polyphenol that exhibits an inhibitory function of inducible nitric oxide synthase.

[23] The dietary supplement according to claim 22, wherein the total weight of the Bacopa monniera extract and olive fruit polyphenol is 5% or more and 98% or less based on the total weight.

P

PPo

[24] The dietary supplement according to any one of claims 18 to 23, further comprising at least one of copper and zinc.

[25] The dietary supplement according to claim 24, wherein copper and zinc are added, and the weight ratio of the added copper and zinc is 1: 1 or more and 1: 200 or less.

[26] Copper and zinc are added, and the weight ratio of added copper to zinc is 1: 4 or more 1: 25. The dietary supplement according to claim 24, wherein the dietary supplement is 100 or less.

27. The dietary supplement according to claim 24, wherein copper and zinc are added, and the weight ratio of the added copper and zinc is 1: 4 or more and 1:50 or less.