JP2016183149A - Oral composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide pharmaceuticals, quasi drugs, food and drink and the like which are highly safe and have NO activity promoting actions.SOLUTION: An nitric oxide activity promoting composition comprises a piperine analog.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a composition for promoting nitric oxide (NO) activity containing a piperine analog.

  NO is a gaseous component produced (synthesized) by nitric oxide synthase (NOS) using L-arginine and molecular oxygen as substrates in vivo. It is regulated by factors.

  NO is considered to be involved in various physiological functions mainly by activating the soluble guanylate cyclase in the target cell and inducing an action via cyclic GMP (Non-patent Document 1). For example, platelet aggregation suppression, anti-arteriosclerotic action and vasodilation, memory and learning functions in the central nervous system, smooth muscle relaxing action through the NO-acting nervous system in the peripheral nervous system, etc. (Non-Patent Document 2).

  For this reason, a decrease in NO synthesis, NO release, and a decrease in reactivity to NO are factors that reduce these functions. Therefore, by enhancing NO synthesis and the like, it can be expected that the physiological function and tissue function of the body related to NO are well maintained or the lowered function is improved.

  For this reason, proposals have already been made for components and compositions that promote NO synthesis and the like. Examples thereof include NO action enhancers such as chlorogenic acid, caffeic acid, ferulic acid (Patent Document 1), hyperoside and isoquercitrin (Patent Document 2). However, the effects of these NO action enhancers are weak and the action specificity for NO is low. For example, in the effect of ferulic acid on the contraction / relaxation function of vascular tissue, the proportion of NO involved is only about 33%. For this reason, a safer component having an excellent NO activity promoting action has been demanded.

  Piperine analogs such as piperine and piperlongminin have anti-tumor activity, antioxidant activity, anti-inflammatory activity, bactericidal activity, anti-gastric ulcer activity, insecticidal activity and other pharmacological effects (Non-patent Document 3), and functional gastroenteropathy. It has been recognized that it has a therapeutic action (Patent Document 3), a pain relieving action (Patent Document 4), a PPARγ activity inhibitory action (Patent Document 5), and the like.

  In addition, piperine and piperlongminin are known to have a blood pressure lowering effect. The blood pressure lowering action of piperine is due to angiotensin converting enzyme (ACE) inhibitory action (Non-patent Document 4), action via voltage-dependent Ca channel (inhibition action of Ca ion inflow into cells) (Non-Patent Document 5).

  Therefore, the promoting action of piperine on NO synthesis and NO release has not been known. Moreover, no activity promoting activity against NOS was known.

Am J Physiol Heart Circ Physiol. 2005; 289 (2): H886-897 Chapter 4 Various actions of NO in living body, P46-56, NO and medicine (physiological and pharmacological action of nitric oxide), Yoshihiko Oyanagi, 1993, Kyoritsu Shuppan Co., Ltd. Phytother Res. 2013; 27 (8): 1121-1130 Indian J Traditional Knowledge 2013; 12 (3): 478-482 J Cardiovasc Pharmacol. 2008; 52 (5): 452-458.

JP2003-261444 JP2008-255075 JP 2006-327999 A JP 2010-280668 A JP2013-237656A

  The present invention relates to providing pharmaceuticals, quasi-drugs, foods and drinks, etc. that are highly safe and have an activity of promoting NO activity.

  As a result of searching for a component that promotes NO activity, the present inventor found that piperine has a NO activity promoting action, and completed the present invention.

That is, the present invention
(1) A composition for promoting nitric oxide activity comprising a piperine analog,
(2) The composition according to (1), which has an action of increasing an enzyme activity or an action of increasing the amount of nitric oxide synthase,
(3) The composition according to any one of (1) to (3), which is a composition according to (1) and (4) a food and drink composition having an action of increasing the amount of nitric oxide synthesis. .

  According to the present invention, it is possible to provide a highly safe composition for promoting NO activity, which contains piperine analogs such as piperine and piperlongminin. The composition of the present invention can favorably maintain the physiological function and tissue function of the body related to NO, or improve the physiological function and tissue function of the body related to the lowered NO.

It is a graph which shows the influence by the vascular endothelial cell removal with respect to the blood vessel relaxation effect | action of piperine (each group 6 cases, the average value of a contraction rate +/- standard error is displayed. The point of Pre shows the contraction rate before piperine treatment. †: p <0.05, ††: p <0.01 vs vascular endothelial cell retention specimen (Student's t-test))

“Promoting nitric oxide (NO) activity” means maintaining NO synthesis and NO release in a normal state, suppressing reduction in NO synthesis and NO release, and improving (increase) NO synthesis and NO release that are decreasing. means. The NO synthesis amount and the NO release amount may be measured using NO as an index, or NO ₂ or NO ₃ which are stable metabolites as an index. Examples of the method for promoting NO activity include a method for increasing the enzyme activity and the amount of NOS and a method for increasing the amount of intracellular NO synthesis. By these methods, the amount of NO synthesis and the amount of NO release increase.

  NOS is classified into constitutive NOS (cNOS) that is always present in a certain amount of cells and inducible NOS (iNOS, NOS2) that is induced by inflammation or stress. Furthermore, cNOS is classified into neural type nNOS (NOS1) and vascular endothelial type eNOS (NOS3), and mitochondrial type mtNOS is also included in cNOS. The NOS in the present invention is preferably cNOS, more preferably eNOS or nNOS, and particularly preferably eNOS. The enzyme activity of NOS can be measured using the amount of intracellular NO synthesis or the degree of NOS phosphorylation as an index, and the amount of enzyme can be measured using the expression level of NOS protein or NOS gene as an index.

  The composition for promoting NO activity in the present invention maintains these functions well with respect to the physiological functions and tissue functions of the body related to NO, improves the decreased functions, and prevents the functions from decreasing. Can be used for the purpose.

Examples of the state in which physiological functions and tissue functions related to NO are lowered include renal dysfunction, increased proteinuria, hematuria, asthma, prolonged cough, gastric mucosal damage, intestinal mucosal damage, erectile dysfunction, and erectile dysfunction , Inability to perform satisfactory sexual intercourse (including insufficient hardness and maintenance), hair loss, reduced cognitive function, decreased concentration, decreased motivation, eye strain, decreased eye function, focus control function Decreased, bear under eyes, hearing loss, neck stiffness, shoulder stiffness, waist beam, back beam, hemoptysis, nosebleed, delayed tissue damage repair, platelet aggregation, thrombus formation, exercise fatigue, muscle weakness, insulin resistance Vascular disorders accompanied by sex, increase in cholesterol, increase in neutral fat, increase in free fatty acid, chilling, etc., and further decreased states include heart failure, coronary artery spasm, worsening of ischemic disease and the like.
Therefore, the composition for promoting NO activity of the present invention can cause renal dysfunction, increased proteinuria, hematuria, asthma, prolonged cough, gastric mucosal damage, intestinal mucosal damage, erectile dysfunction, erectile dysfunction, and satisfactory sexual intercourse. No state (including insufficient hardness and maintenance), hair loss, cognitive decline, poor concentration, motivation, eye strain, reduced eye function, poor focus control, bear under eyes It can be used as a composition for prevention or improvement of hearing loss, neck stiffness, shoulder stiffness, waist beam, back beam, hemoptysis, nosebleed, delayed repair of tissue damage, and the like.

The composition of the present invention contains a piperine analog.
“Piperine analog” means a compound similar in molecular biological properties such as chemical structure and receptor binding properties to piperine, preferably piperine, piperlongminin, pipernonalin, Contains one or more components selected from dihydropiperine, dihydropiperlongminin, and dihydropipernonalin. Piperine or piperlongminin is more preferable, and piperine is more preferable. These piperine analogs are particularly preferably contained as active ingredients.

  These piperine analogs are commercially available as chemically synthesized products, but may be synthesized from piperidine derivatives or the like, or may be extracts from natural products containing piperine analogs.

  The natural product containing a piperine analog is, for example, a natural product belonging to the genus Pepperaceae such as black pepper. Extraction from natural products is performed by using a distillation method such as steam distillation in addition to solvent extraction performed by impregnation in a solvent at room temperature or in a heated state or using an extraction device such as a Soxhlet extractor, A supercritical extraction method in which carbon dioxide gas is in a supercritical state, or a pressing method for obtaining an extract by pressing can be used.

  As the extraction solvent used for the solvent extraction, either a polar solvent or a nonpolar solvent can be used, for example, water, ethanol or hexane, which can be used alone or in combination of two or more. It is also possible to repeat the process by changing the solvent. Of these, water and ethanol are preferably used, and water is more preferably used. Examples of the means for separating and purifying the extract include activated carbon treatment, liquid-liquid distribution, column chromatography, liquid chromatography, gel filtration, and precision distillation.

  The extract containing the piperine analog used in the present invention may be the extract or separated and purified product as it is, may be used as a diluted solution diluted with an appropriate solvent, may be used as a concentrated extract or dry powder, or paste What was prepared in the shape may be sufficient.

  The total intake amount of piperine analog per day can be appropriately increased or decreased in consideration of age, sex, weight, etc., but is preferably 0.01 mg to 100 mg, more preferably 0.02 mg to 20 mg, more preferably 0.04 mg 2 mg is more preferred.

  The daily intake of piperine is preferably 0.01 mg to 50 mg, more preferably 0.02 mg to 10 mg, and particularly preferably 0.04 mg to 1 mg.

  The daily intake can be administered (taken) once a day or divided into several times.

  The composition of the present invention can contain vitamins, amino acids, minerals, herbal medicines, extracts thereof, and the like as long as the effects of the present invention are not impaired in addition to piperine analogs. Then, excipients, disintegrants, binders, lubricants, antioxidants, coating agents, coloring agents, flavoring agents, etc. are blended, and pH adjusters, cooling agents, suspending agents are added as necessary. Oral solid preparations or drinks such as tablets, capsules, powders, granules, dry syrups, etc. by blending agents, antifoams, thickeners, solubilizers, surfactants, fragrances, etc. It can be provided as an internal liquid for beverages, concentrated beverages, solid sparkling beverages, powdered beverages and the like. In addition, as a state of the piperine analog in an internal use liquid agent, it may be a dissolved state or a dispersed state, and the presence state is not ask | required.

  The composition for promoting nitric oxide activity of the present invention is usually provided as a pharmaceutical, quasi-drug, etc. that can be used for efficacy and effect. Moreover, it is also possible to provide food and drink such as functional display food, food for specified health use, and nutritional functional food. As a product that displays the indications of efficacy and effects that can be expected by ingestion, in the body's physiological functions and tissue functions related to NO, products with notation that mean that these functions are well maintained and that reduced functions are improved Is mentioned. For example, renal dysfunction, increased proteinuria, hematuria, asthma, prolonged cough, gastric mucosal injury, intestinal mucosal injury, erectile dysfunction, erectile dysfunction, inability to perform satisfactory sexual intercourse (hardness and maintenance are insufficient Hair loss, reduced cognitive function, reduced concentration, reduced motivation, eye strain, decreased eye function, decreased focus control, bear under eyes, hearing loss, neck stiffness, shoulder stiffness, It is also possible to provide a label indicating that the waist beam, back beam, hemoptysis, nosebleed, delay in repairing tissue damage, etc. are improved.

  When the composition of the present invention is used as a food or drink, specific examples of the food or drink include liquid foods such as drinks, soups, milk drinks, soft drinks, tea drinks, alcoholic drinks, jelly drinks, and functional drinks. Products containing oil such as edible oil, dressing, mayonnaise, margarine; foods containing carbohydrates such as rice, noodles and bread; processed livestock foods such as ham and sausage; processed fishery products such as kamaboko, dried fish and salted fish; pickles Processed vegetable foods such as semi-solid foods such as jelly and yogurt; Fermented foods such as miso and fermented beverages; Various confectioneries such as Western confectionery, Japanese confectionery, candy, gums, gummy, frozen confectionery, ice confectionery; Examples include retort products such as ankake and Chinese soup; instant foods such as instant soup and instant miso soup and foods compatible with microwave ovens. Furthermore, health foods and drinks prepared in the form of powder, granules, tablets, capsules, liquid, paste or jelly are also included. Manufacture of the food-drinks in this invention can be implemented with a manufacturing technique well-known in the said technical field.

  Hereinafter, representative test examples and examples of the present invention will be shown and described in detail.

1. Evaluation of NO activity promoting action using rat thoracic aorta (test method)
For the experiment, male SD rats (Crl: CD, Nippon Charles River Co., Ltd.) of 8 to 12 weeks old were used. Piperine (manufactured by Wako Pure Chemical Industries, Ltd.) was used as a representative piperine analog.

  Rats were euthanized after anesthesia and the thoracic aorta was removed. After removing the connective tissue and adipose tissue attached to the thoracic aorta, a vascular ring specimen having a width of about 3 mm was prepared and used as a vascular endothelial cell holding specimen. The vascular endothelial cell-removed specimen was prepared by passing a cotton thread through the lumen of the vascular specimen and peeling the vascular endothelial cells by friction.

  The piperine used in the test was dissolved in DMSO, the vasoconstrictor phenylephrine, the vasorelaxant acetylcholine, the NOS inhibitor L-NAME, and the COX inhibitor indomethacin in water for injection.

The Krebs solution heated to 37 ° C. was filled in an organic bath, and 95% O ₂ /5% CO ₂ was aerated. The blood vessel specimen was suspended in an organ bath with a tension of 1 g.
The tension of the blood vessel specimen was measured using a polygraph system via an FD pickup, a sensor coupler, and a sensor amplifier. As the vascular endothelial cell-retaining specimen, a specimen in which a relaxation reaction due to acetylcholine (10 μmol / L) is markedly observed after observing phenylephrine (1 μmol / L) -induced vasoconstriction reaction is selected. Specimens with a relaxed relaxation response were selected.

(Test 1)
After confirming the vascular endothelial cell-dependent vasorelaxation reaction of the vascular specimen, the specimen was washed again to induce a vasoconstriction reaction by phenylephrine, and the vasorelaxing action of piperine was evaluated. Verification of the action of piperine on vascular endothelial cells was performed by comparing the vasorelaxant action of piperine using a vascular endothelial cell-retaining specimen and a vascular endothelial cell-removed specimen.
(Test 2)
The verification of the action of piperine through NOS was conducted by evaluating the effect of L-NAME (100 μmol / L) or indomethacin (10 μmol / L) pretreatment on the relaxation action of piperine using vascular endothelial cell-retaining specimens. I went.

The relaxation effect of piperine was evaluated using the contraction rate calculated from the following equation.
Shrinkage rate (%) = (Shrinking tension after adding piperine / Shrinking tension before adding piperine) × 100

(Result 1)
The result of verifying the action of piperine on vascular endothelial cells is shown in FIG.
The effect of vascular endothelial cell removal on the vasorelaxant effect of piperine was evaluated. As a result, 10 μmol / L to 100 μmol / L piperine suppresses phenylephrine-induced vasoconstriction with increasing concentration, and the contraction rate of 100 μmol / L piperine is 11.8% (average value) )Met. On the other hand, in the vascular endothelial cell-removed specimen, the contraction rate of 100 μmol / L piperine was 70.0%, and the relaxation action of piperine was remarkably attenuated (p <0.05, piperine in the specimen containing vascular endothelial cells). Shrinkage rate (Student's t test), number of cases 6).
Thus, it has been clarified that piperine acts on vascular endothelial cells, unlike the previous findings.

(Result 2)
The result of having verified the effect | action through the NOS of piperine is shown.
The effect of L-NAME on the vasorelaxant effect by piperine was evaluated using a vascular endothelial cell-retaining specimen. As a result, the shrinkage of 100 μmol / L piperine in L-NAME untreated was 37.2%, but the shrinkage of 100 μmol / L piperine in L-NAME pretreatment was 76.9% ( p <0.05 vs. piperine contraction rate when untreated with L-NAME (Student's t test), number of cases 6).
On the other hand, the contraction rate of 100 μmol / L piperine in indomethacin pretreatment was 32.2%, and the contraction rate of 100 μmol / L piperine in indomethacin untreated simultaneously was 40.8%.
Accordingly, the ratio of the effect of NO in the action of piperine is about 63% (relaxation rate 39.7 attenuated by L-NAME pretreatment accounting for the relaxation rate of piperine 62.8% <100% -37.2%>). % <76.9% -37.2%>).

  From the above test examples, it has been clarified that piperine exhibits an effect through synthesis of NO by eNOS.

2. Evaluation of increase in intracellular NO synthesis using human-derived vascular endothelial cells (test method)
In the experiment, human-derived vascular endothelial cells (HUVEC: Kurashiki Boseki Co., Ltd.) were used. Human-derived vascular endothelial cells were used for the test after culturing to subconfluence using a subculture reagent set.

  As a typical piperine analog, piperine (manufactured by Wako Pure Chemical Industries, Ltd.) was used. Piperine was dissolved in DMSO and used for testing.

  The NO activity promoting action was evaluated using an increase in intracellular NO amount as an index, and the intracellular NO amount was measured using the fluorescent dye DAF-FM DA.

(Test 3)
To incorporate DAF-FM DA into the cells, the cells were treated with a medium containing DAF-FM DA for 60 minutes. Thereafter, the medium was replaced with a medium containing piperine (100 μmol / L) and treated for 60 minutes.
Cells were detached from the culture vessel using Trypsin solution, suspended in PBS solution, and fluorescence intensity of DAF-FM DA per living cell was measured using a flow cytometer (EPICS XL, manufactured by Beckman Coulter, Inc.). .

(Result 3)
The effect of piperine on the amount of NO synthesis in human-derived vascular endothelial cells was evaluated. As a result, the addition of 100 μmol / L piperine significantly increased the amount of intracellular NO synthesis by 23.8% (mean value) compared to the case without piperine (p <0.05 vs. cells without piperine). NO synthesis amount (Wilcoxon test), number of cases 7) each.
From this, it was revealed that piperine increases the amount of NO synthesis in human vascular endothelial cells and has a NO activity promoting action.

  Therefore, piperine analogs such as piperine and piperlongminin can be used as a composition for promoting nitric oxide activity, and these functions are exerted on the physiological functions and tissue functions of the body related to NO. It has been clarified that it has an effect of maintaining good, improving a lowered function, and preventing a decrease in function.

  Compositions containing piperine analogs such as piperine and piperlongminin of the present invention are useful as pharmaceuticals for promoting nitric oxide activity, quasi-drugs, foods indicating that they have functionality, etc. . Moreover, as a composition useful for maintaining these functions well with respect to the physiological functions and tissue functions of the body related to NO, improving the decreased functions, and preventing the decrease in the functions, It became possible to ingest.

Claims (4)

A composition for promoting nitric oxide activity, comprising a piperine analog. The composition according to claim 1, which has an effect of increasing the enzyme activity or the amount of enzyme of nitric oxide synthase. The composition according to claim 1, which has an effect of increasing the amount of nitric oxide synthesis. The composition of any one of Claims 1-3 which is a food-drinks composition.

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