JP2007186431A - Antioxidation composition, cerebral nerve cell-protecting pharmaceutical composition, antioxidation agent, cerebral nerve cell-protecting agent and use thereof - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for suppressing oxidation in the body, and a composition, medicine and food for oxidation suppression use. <P>SOLUTION: Krachai (Boesenbergia rotunda) is processed to dried material, powder or extract or 4-hydroxypanduratin is purified, and the product is used as a component of a composition, medicine, food composition or food. Administration of these products to human or non-human vertebrates enables the suppression of oxidation in the body, the suppression of oxidation by active oxygen/radical in the body and the suppression of cerebral nerve cell disorder caused by excessive intake of glutamic acid. Perce Pierre is processed to dried material, powder or extract, and the product is used as a component of a composition, medicine, food composition or food. Administration of these products to human or non-human vertebrates enables the suppression of oxidation in the body and the suppression of oxidation by active oxygen/radical in the body. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to an antioxidant composition containing a processed product of Krachai and a pharmaceutical composition for protecting cranial nerve cells, an antioxidant containing 4-hydroxypanduratin as an active ingredient, a cranial nerve cell protecting agent, and an antioxidant containing processed Paspierre. The present invention relates to compositions and methods for their use.

Active oxygen is deeply involved in morbidity of various lifestyle-related diseases, aging, cancer and the like. In general, it is considered that active oxygen is naturally generated in the body by stimulation of air pollution due to environmental pollution, radiation, foods containing additives, smoking, ultraviolet rays, stress and the like. Further, it is considered that about 2% of oxygen taken into the body by respiration becomes active oxygen, and this active oxygen damages the cells of the body (for example, see Non-Patent Documents 1 and 2).
“Kunio Yagi, Atsushi Nakano (supervised), Akio Futaki, Hiroyuki Shimazaki (ed.):“ Reactive Oxygen Chemistry, Biology, Medicine ”, Medical and Dental Publishing (1987) "Halliwell, B., and Gutteridge, JMC (translated by Mitsuyoshi Matsuo, Masaru Sakurai, Toshikazu Yoshikawa):" Free radicals and living organisms ", Academic Publishing Center (1988)"

In this way, the factors that generate active oxygen are deeply involved in our living environment, and we are always subject to active oxygen.
Then, an object of this invention is to provide the method for suppressing the body oxidation, and the composition, chemical | medical agent, and foodstuff for it.

  As shown in the following examples, the present inventors have studied a novel plant having an antioxidant action, and found that it has an antioxidant action on Krachai and Paspierre. Furthermore, as a result of analyzing compounds contained in Krachai, it was found that 4-hydroxypanduratin has a strong antioxidant action and cranial nerve protective action, and the present invention has been completed.

  That is, the antioxidant composition according to the present invention is characterized by containing a processed Krachai product. Here, the processed product is preferably, for example, a dried product, a powder, or an extract.

  The composition according to the present invention is a composition for reducing oxidation by active oxygen / radicals, and is characterized by containing a processed product of Krachai. Here, the processed product is preferably, for example, a dried product, a powder, or an extract.

  Furthermore, the composition for protecting cranial nerve cells according to the present invention is a composition for protecting cranial nerve cells for suppressing cranial nerve cell damage caused by excessive intake of glutamic acid, and contains a processed product of Krachai. . Here, the processed product is preferably, for example, a dried product, a powder, or an extract.

  Moreover, the food composition according to the present invention is a food composition having an antioxidative action, and is characterized by containing a processed product of Krachai. Here, the food composition is characterized by constituting, for example, health food, functional food, or health functional food.

  Furthermore, the food according to the present invention is characterized in that it contains a processed product of Krachai, has an antioxidant inhibitory action, and is a food with an indication that it is used for suppressing oxidation in the body. Features.

  Further, the food according to the present invention is characterized in that it contains a processed product of Krachai, has an antioxidant inhibitory action, and is used for treatment, prevention, or alleviation of symptoms due to oxidation in the body. It is characterized by being a food with a label.

  Furthermore, the food composition according to the present invention is a food composition that reduces oxidation by active oxygen / radicals, and is characterized by containing a processed product of Krachai. Here, the food composition is characterized by constituting, for example, health food, functional food, or health functional food.

  In addition, the food according to the present invention contains a processed product of Krachai, is characterized by reducing oxidation by active oxygen / radicals, and is used for reducing oxidation by active oxygen / radicals present in the body. It is characterized by being a food with a sign to the effect.

  Furthermore, the food composition according to the present invention is a food composition for suppressing cerebral nerve cell damage due to excessive intake of glutamic acid, and is characterized by containing a processed product of Krachai. Here, the food composition is characterized by constituting, for example, health food, functional food, or health functional food.

  The food according to the present invention contains a processed product of Krachai, is characterized by suppressing cranial nerve cell damage caused by excessive intake of glutamic acid, and is used for suppressing cranial nerve cell damage caused by excessive intake of glutamic acid. It is characterized by being a food with a sign to the effect.

  Furthermore, the food according to the present invention contains a processed product of Krachai, and is characterized by suppressing cranial nerve cell damage caused by excessive intake of glutamic acid, for the treatment, prevention, or alleviation of symptoms of neurological diseases caused by excessive intake of glutamic acid. It is characterized by being a food with an indication that it is used in

  The antioxidant according to the present invention is characterized by containing 4-hydroxypanduratin as an active ingredient.

  Furthermore, the drug according to the present invention is a drug that reduces oxidation by active oxygen / radicals, and is characterized by containing 4-hydroxypanduratin as an active ingredient.

  The cerebral nerve cell protective agent according to the present invention is a cerebral nerve cell protective agent for suppressing cerebral nerve cell damage due to excessive intake of glutamic acid, and is characterized by containing 4-hydroxypanduratin as an active ingredient.

  Furthermore, the food composition according to the present invention is a food composition having an antioxidant action, and is characterized by containing 4-hydroxypanduratin. Here, the food composition is characterized by constituting, for example, health food, functional food, or health functional food.

  In addition, the food according to the present invention is a food containing 4-hydroxypanduratin, having an antioxidant inhibitory action, and having a label indicating that it is used to suppress in-vivo oxidation. It is characterized by being.

  Furthermore, the food according to the present invention is characterized by containing 4-hydroxypanduratin, having an antioxidant inhibitory effect, and used for the treatment, prevention or alleviation of symptoms caused by internal oxidation. It is characterized by being a food with a label to that effect.

  The food composition according to the present invention is a food composition that reduces oxidation by active oxygen / radicals and is characterized by containing 4-hydroxypanduratin. Here, the food composition is characterized by constituting, for example, health food, functional food, or health functional food.

  Furthermore, the food according to the present invention is characterized by containing 4-hydroxypanduratin and reducing oxidation by active oxygen / radical, and is used for reducing oxidation by active oxygen / radical present in the body. It is characterized by being a food with a label indicating that it is a product.

  The food composition according to the present invention is a food composition that suppresses cranial nerve cell damage due to excessive intake of glutamic acid, and is characterized by containing 4-hydroxypanduratin. Here, the food composition is characterized by constituting, for example, health food, functional food, or health functional food.

  Furthermore, the food according to the present invention contains 4-hydroxypanduratin, is characterized by suppressing cranial nerve cell damage due to excessive intake of glutamic acid, and is labeled as being used to protect cranial nerve cells. It is a food product.

  Further, the food according to the present invention contains 4-hydroxypanduratin, is characterized by suppressing cerebral nerve cell damage caused by excessive intake of glutamic acid, for the treatment, prevention, or alleviation of symptoms of neurological diseases caused by excessive intake of glutamic acid. It is characterized by being a food with an indication that it is used in

  Furthermore, the antioxidant composition according to the present invention is characterized by containing a processed Pastier product. Here, the processed product is preferably, for example, a dried product, a powder, or an extract.

  Moreover, the composition concerning this invention is a composition which reduces the oxidation by active oxygen / radical, Comprising: It is characterized by containing a Paspierre processed material. Here, the processed product is preferably, for example, a dried product, a powder, or an extract.

  Furthermore, the food composition according to the present invention is a food composition having an antioxidative action, and is characterized by containing a processed Pastier product. Here, the food composition is characterized by constituting, for example, health food, functional food, or health functional food.

  Further, the food according to the present invention is a food containing a processed pastier product, having an antioxidant inhibitory effect, and having a label indicating that it is used to suppress internal oxidation. It is characterized by.

  Furthermore, the food according to the present invention is characterized in that it contains a processed pastier product, has an antioxidant inhibitory effect, and is used for treatment, prevention, or alleviation of symptoms due to oxidation in the body. It is characterized by being a food with the label.

  The food composition according to the present invention is a food composition that reduces oxidation by active oxygen / radicals, and is characterized by containing a processed pastier product. Here, the food composition constitutes a health food, a functional food, or a health functional food.

  Furthermore, the food according to the present invention contains a processed Paspierre, is characterized by reducing oxidation by active oxygen / radical, and is used to reduce oxidation by active oxygen / radical present in the body. It is characterized by being a food with a label to that effect.

  Here, Krachai is Boesenbergia pandurata, but also includes Kaempferia rotunda, Boesenbergia rotunda, Gastrochilus panduratus, and Kaempferia pandurata, which are known as synonyms.

  According to the present invention, it is possible to provide a method for suppressing body oxidation, and a composition, drug, and food for the same.

  Hereinafter, the present invention will be described specifically and in detail with reference to examples, but the present invention is not limited thereto.

  The objects, features, advantages, and ideas of the present invention will be apparent to those skilled in the art from the description of the present specification, and those skilled in the art can easily reproduce the present invention from the description of the present specification. it can. The embodiments and specific examples of the invention described below show preferred embodiments of the present invention, and are shown for illustration or explanation. It is not limited. It will be apparent to those skilled in the art that various modifications can be made based on the description of the present specification within the spirit and scope of the present invention disclosed herein.

=== Production Method of Krachai Processed Product and Pastier Processed Product ===
Krachai used for processed Krachai (scientific name: Boesenbergia pandurata) and Paspierre used for processed Papierre (scientific name: Salicornia europaea L.) are either wild or sold at general grocery stores. But you can. Here, examples of the processed product include, but are not limited to, a dried product, a powder, and an extract. The manufacturing method of a processed material is as follows, for example.

  The dried product can be produced by air drying, freeze drying or the like. The powder can be obtained by cutting or grinding the above-mentioned dried product using, for example, a mixer. The extract is extracted with chloroform / methanol (for example, chloroform: methanol = 3: 1 to 1: 1 solution), ethanol extraction (for example, 60% to 100% ethanol is used), methanol extraction (for example, 60% ˜100% methanol), water extraction (eg using 60-100 ° C. hot water).

=== 4-Hydroxypanduratin Extraction Method ===
Although the extraction method of 4-hydroxypanduratin is not specifically limited, For example, the following method is mentioned.
First, krachai is lyophilized and powdered through a mixer. This powder is extracted in a solution of CH ₂ Cl ₂ -MeOH (1: 1). The extract is filtered, and the filtrate is concentrated to dryness under reduced pressure. This is applied to a silica gel column equilibrated with hexane-EtOAc (5: 1) and developed with the same solvent. Finally, whether or not the developed product is (-)-4-hydroxypanduratin is confirmed by ESI-MS, NMR analysis (including two-dimensional NMR) and the like.

=== Antioxidant Composition ===
In general, oxidative stress is induced when the balance between the oxidation reaction that is indispensable for a living organism to maintain a life phenomenon and the antioxidant defense mechanism that inactivates the excessively produced active oxygen is lost. Causes a lipid peroxidation reaction.

  As found by the inventors, as described in the following examples, the Krachai processed product, 4-hydroxypanduratin, and the Paspierre processed product inhibit lipid peroxidation in cells. Therefore, compositions containing Krachai processed products and 4-hydroxypanduratin, and compositions containing Paspierre processed products as active ingredients are useful as antioxidant compositions.

  As described above, the Krachai processed product, 4-hydroxypanduratin, and the Paspierre processed product inhibit lipid peroxidation. Since cell membrane lipids contain a large amount of unsaturated fatty acids, they are susceptible to oxidative attack by active oxygen and free radicals, resulting in lipid peroxidation. Therefore, the composition containing the processed Krachai product, the composition containing 4-hydroxypanduratin as an active ingredient, and the composition containing the processed Paspierre are also useful as a composition for reducing oxidation by active oxygen / radicals. is there.

The active oxygen means an oxygen molecule and its related substance in an activated state rather than a normal oxygen molecule (O ₂ ), and includes radical species <superoxide (· O ₂ ⁻ ), hydroxyl radical ( HO.), Hydroperoxyl radical (HOO.), Alkoxyl radical (LO.), Alkylperoxyl radical (LOO.), Nitric oxide (NO) etc.> and non-radical species <hydrogen peroxide (H ₂ O ₂ ), Singlet oxygen ( ¹ O ₂ ), peroxynitrite (ONOO ⁻ ), lipid hydroperoxide (LOOH), hypochlorous acid (HOCl), ozone (O ₃ ) and the like.

  A radical refers to an atom or molecule that has a strong oxidizing action and easily oxidizes by taking one electron from another molecule, and often has an unpaired electron.

  Here, active oxygen / radical means active oxygen and / or a radical, which has an action mechanism of easily oxidizing a partner molecule, and oxidizes phospholipids of lipids, particularly cell membranes, It is a molecular species that damages blood vessels by oxidative damage to proteins and DNA, and promotes aging and canceration. Reducing oxidation by active oxygen / radicals not only prevents adult diseases, aging, and canceration, but also has an effect of suppressing their progression.

=== Antioxidant ===
As the pharmaceutically acceptable carrier that can be used in the antioxidant containing the antioxidant composition of the present invention, various organic or inorganic carrier substances that are commonly used as pharmaceutical materials may be used. Agents, lubricants, binders and disintegrants, or solvents in liquid preparations, solubilizers, suspending agents, isotonic agents, buffers, soothing agents, and the like may be contained. Further, if necessary, additives such as usual preservatives, antioxidants, colorants, sweeteners, adsorbents, wetting agents and the like may be appropriately contained. As the dosage form, oral preparations include, for example, tablets, capsules, granules, powders, fine granules, syrups, sustained-release tablets / capsules / granules, cashews, chewable tablets or drops. Examples of injections include solution injections, emulsion injections, and solid injections.

  The dosage of the antioxidant of the present invention varies depending on age, body weight, indication, or administration / intake route, but is not particularly limited as long as the above-described effects can be exerted and the side effects produced are within an acceptable range. .

  In addition, as a criterion for determination of administration, for example, in a human or non-human vertebrate, a blood peroxide concentration (for example, lipid peroxide) is measured, and if the value is equal to or higher than a predetermined value, the Krachai process Or 4-hydroxypanduratin, and processed Paspierre. Here, the “predetermined value” is, for example, a value that is generally determined that the peroxide concentration in the blood (for example, lipid peroxide) is higher than that of a healthy person, a reference value that is determined by a medical worker, etc. Can be considered. Specific methods for measuring lipid peroxides include the thiobarbituric acid (TBA) method or the hemoglobin-methylene blue method.

  Or, for example, diseases caused by oxidation in the human or non-human vertebrates (eg, arteriosclerosis, myocardial infarction, cerebral infarction, diabetes, retinopathy of prematurity, liver disease, aging, nerve cell disorder, cancer, etc.) In the case of suffering from or when it is necessary to prevent the disease, there may be a case where a processed Krachai product or 4-hydroxypanduratin and a processed Pastier product are administered.

  In addition, “diseases caused by internal oxidation” include arteriosclerosis, myocardial infarction, cerebral infarction, diabetes, retinopathy of prematurity, liver disease, aging, nerve cell disorder, cancer, etc., but oxidation or active oxygen / If it is a disease resulting from oxidation by radicals, it is not limited to these.

  In addition to natural antioxidants such as carotenoids and vitamins C and E, this agent is included in herbal medicines such as polyphenols such as flavonoids and tannins, caffeic acid derivatives, lignans, and saponins in order to enhance antioxidant activity. It may be used in combination with antioxidants.

  Furthermore, this agent has a function of reducing oxidation by active oxygen / radical. Therefore, it is expected that the effect of this drug will be enhanced if this drug is used in combination with an in vivo radical scavenging substance such as SOD (superoxide dismutase), catalase, vitamin E, or vitamin C.

=== Cranial nerve cell protective composition ===
When glutamic acid is administered to cultured nerve cells, nerve cell damage such as nerve cell death occurs. It has also been reported that when mice are given excess glutamate orally or parenterally to neonates, the arcuate nucleus and retina of the hypothalamus are altered peculiarly. The risk of disability and brain development disorders has been pointed out.

  In addition, clinically, there is a disease called Chinese food syndrome, and excessive intake of sodium glutamate is a neurological disease or neurological disorder such as nausea, headache, dizziness, nausea, and limb numbness, even for adults. It is known that symptoms associated with the symptom occur.

  As shown in the following examples, 4-hydroxypanduratin can suppress neuronal cell damage caused by excessive administration of glutamic acid. Therefore, a composition containing 4-hydroxypanduratin or a processed product of Krachai rich in it as an active ingredient is useful as a cranial nerve cell protective composition.

=== Cranial nerve cell protective agent ===
In the cerebral nerve cell protective agent containing the cerebral nerve cell protective composition of the present invention, the pharmaceutically acceptable carrier, administration route and dosage are as described in the above “antioxidant”.

  In addition, as a criterion for administration, for example, in humans or non-human vertebrates, the concentration of glutamic acid in blood or in the brain is measured, and when the value is a predetermined value or more, 4-hydroxypanduratin or Krachai processed product Is administered. Here, the “predetermined value” may be, for example, a value that is generally determined that the concentration of glutamic acid in the blood or brain is higher than that of a healthy person, or a reference value determined by a medical worker.

  Or, if you suffer from a neurological disease (eg, brain cell damage, neuropathy) caused by excessive intake of glutamic acid, or if you need to prevent the disease, administer 4-hydroxypanduratin or Krachai processed product. There may be cases.

== Processed Krachai or food composition or food containing 4-hydroxypanduratin ==
Giving a human or non-human vertebrate food composition or food containing 4-hydroxypanduratin includes, for example, cranial nerves caused by oxidation in the body, oxidation by active oxygen / radicals, or excessive intake of glutamate It enables the treatment, prevention, or alleviation of symptoms resulting from cell damage.

  In addition, these food compositions or foods are “used to suppress oxidation in the body”, “used to treat, prevent or alleviate symptoms caused by oxidation in the body”, “exist in the body” Used to reduce oxidation by active oxygen / radicals "," used to suppress cerebral nerve cell damage caused by excessive intake of glutamate ", or" treatment, prevention of neurological diseases caused by excessive intake of glutamate, Or, it is preferable to add an indication of “used for symptom relief”.

  Here, the “disease caused by internal oxidation” includes, for example, arteriosclerosis, myocardial infarction, cerebral infarction, diabetes, retinopathy of prematurity, liver disease, aging, nerve cell disorder, cancer, etc. As long as it is a disease caused by oxidation by active oxygen / radical, it is not limited thereto.

  When a food composition or food containing Krachai processed product or 4-hydroxypanduratin is taken orally, it may be in any form such as solid or liquid.

  The intake of the food composition or food of the present invention varies depending on the age, weight, indication, or administration / intake route, but it is caused by oxidation of the body, oxidation by active oxygen / radical, or cranial nerve cell damage due to excessive intake of glutamate. There is no particular limitation as long as it can be treated, prevented, or alleviates symptoms, and the side effects that occur can be tolerated.

  In addition, the food composition or food of the present invention has a natural antioxidant composition such as carotenoid and vitamin C · E, as well as polyphenols such as flavonoids and tannins, caffeic acid derivatives and lignans in order to enhance the antioxidant activity. -You may use together with the other antioxidant contained in crude drugs, such as saponins.

  Furthermore, the food composition or food of the present invention also has a function of reducing oxidation by active oxygen / radical. Therefore, in order to further enhance the effect of the food composition or food of the present invention, the food composition or food of the present invention and an in vivo radical scavenging substance such as SOD (superoxide dismutase), catalase, vitamin E, or vitamin C are used. You may use together.

  In addition, it is said that when a large amount of sodium glutamate, which is known as a seasoning, is ingested at a time, cranial nerve cell damage is caused. Therefore, the food composition or food of the present invention is suitable for eating with glutamic acid (for example, sodium glutamate).

== Food composition or food containing processed Pastiers ==
Giving a human or non-human vertebrate a food composition or food containing a processed Paspierre, for example, treatment, prevention of oxidation in the body or oxidation by active oxygen / radicals, or symptoms resulting therefrom Allows relaxation.

  In addition, these food compositions or foods are “used to suppress oxidation in the body”, “used to treat, prevent or alleviate symptoms caused by internal oxidation”, or “ It is preferable to indicate that it is “used to reduce oxidation by active oxygen / radicals present”.

  Here, the “disease caused by internal oxidation” includes, for example, arteriosclerosis, myocardial infarction, cerebral infarction, diabetes, retinopathy of prematurity, liver disease, aging, nerve cell disorder, cancer, etc. As long as it is a disease caused by oxidation by active oxygen / radical, it is not limited thereto.

  When the food composition or food containing the processed Pastier product is taken orally, it may be in any form such as solid or liquid.

  The intake of the food composition or food of the present invention varies depending on the age, body weight, indication, or administration / intake route, but the suppression of internal oxidation or the treatment, prevention, or oxidation of active oxygen / radical There is no particular limitation as long as the symptoms caused can be alleviated and the side effects that occur are within an acceptable range.

  In addition, the food composition or food of the present invention has a natural antioxidant composition such as carotenoid and vitamin C · E, as well as polyphenols such as flavonoids and tannins, caffeic acid derivatives and lignans in order to enhance the antioxidant activity. -You may use together with the other antioxidant contained in crude drugs, such as saponins.

  Furthermore, the food composition or food of the present invention also has a function of reducing oxidation by active oxygen / radical. Therefore, in order to further enhance the effect of the food composition or food that reduces oxidation by active oxygen / radicals, the food composition or food of the present invention and SOD (superoxide dismutase), catalase, vitamin E, vitamin C, etc. An in vivo radical scavenger may be used in combination.

  Hereinafter, although the embodiment described above will be specifically described using examples, this is an exemplification, and the present invention is not limited to these examples. In the following examples, Boesenbergia pandurata was used as Krachai.

<Example 1: Experiment using Krachai processed product and 4-hydroxypanduratin>
=== Preparation of Antioxidant Active Substance ===
(1) Production of Krachai powder 1.5 kg of Krachai (commercial product) was freeze-dried and powdered with a mixer. Using the same method, tea leaves (commercial product: Jurakuen “Ichikai no Tomo”) and ginkgo grass (commercial product) were also powdered.

(2) Preparation of Krachai extract Chloroform / methanol extraction (chloroform: methanol = 1: 1), ethanol extraction (use 100% ethanol), methanol extraction (use 100% methanol), hot water extraction (heat at 100 ° C) Krachai extract was obtained by using water). A specific extraction method is as follows.

First, 1.5 kg of Krachai was lyophilized and powdered with a mixer. Of these, 16 g each was (i) 100 ml solution of CH ₂ Cl ₂ -MeOH (1: 1), (ii) 100 ml EtOH solution, (iii) 100 ml MeOH solution, (iv) 100 ml hot water at 100 ° C, ( v) Extracted twice with 100 ml of water at 20 ° C. The extract was filtered and the filtrate was concentrated to dryness under reduced pressure.The weights were (i) 1.4 g, (ii) 1.0 g, (iii) 1.6 g, (iv) 2.6 g, (v) 2.0 g, respectively. Met.
Moreover, the tea leaf extract was also produced using the method similar to the above.

(3) Isolation of 4-hydroxypanduratin Krachai 1.5 kg (purchased) was freeze-dried and powdered with a mixer. This _{CH 2 Cl 2 -MeOH (1:} 1) solution 1L of and extracted twice. The extract was filtered, and the filtrate was concentrated to dryness under reduced pressure. The weight was 2.89 g. This was applied to a silica gel column (diameter 3 cm × length 20 cm, silica gel 60 (Merck)) equilibrated with hexane-EtOAc (5: 1) and developed with the same solvent.

  As a result, six kinds of compounds as an active substance were eluted as a single compound. As a result of conducting NMR analysis including ESI-MS and 2D NMR for each compound, 5-hydroxy-7-methoxyflavanone (458.0mg), (-)-panduratin A (402.5mg), 5,7-dihydroxyflavanone, respectively (270.1mg), 2 ', 6'-dihydroxy4'-methoxychalcone (47.0mg), 2', 4'-dihydroxy-6'-methoxychalcone (39.1mg), (-)-4-hydroxypanduratin A (213.2mg) Identified.

The MS and NMR data that led to the identification of (-)-4-hydroxypanduratin A are as shown below. Here, the number of carbon in NMR is added to the structural formula.
ESI-MS 393 (M + H) ⁺ , 415 (M + Na) ⁺

本データは、以前に報告された(-)-4-hydroxypanduratin Aのデータ(J. Agric. Food Chem., vol. 49 (6), 3046と一致した。 <400 MHz ¹ H NMR (in C ₆ D ₆ ) and ¹³ C NMR data of (-)-4-hydroxypanduratin A (shown in DMSO-d ₆ )>

This data was consistent with previously reported data for (-)-4-hydroxypanduratin A (J. Agric. Food Chem., Vol. 49 (6), 3046).

=== Rat brain lipid peroxidation inhibitory experiment ===
In the following experiment, frozen rat brain (Funakoshi, catalog No. J-201) was used after being homogenized under ice cooling. Moreover, since tea leaves have an antioxidant effect, they were used as a positive control, and Ginkgo grass was used as a negative control because it did not have an antioxidant effect. The experimental method is as follows.

(1) Rat brain lipid peroxidation inhibitory experiment using Krachai processed product
In 0.6 ml of 100 mM phosphate buffer (pH 7.4), the test sample (Krachai powder 8 μg / ml to 4 mg / ml and Krachai extract (concentration is 32 μg / ml to 16 mg in terms of powder amount, respectively). 0.05 ml, 0.1 ml of 1 mM ascorbic acid and 0.05 ml of distilled water were added and preincubation was performed for 5 minutes at 37 ° C. Here, the method of Kubo et al (Cheeseman, KH; Forni, LG An investigation of the novel anti-inflammatory agents ONO-3144 and MK-447. Studies on their potential antioxidant activity. 0.2 ml of rat brain homogenate (2.5% (w / v)) prepared by Biochem Pharmacol, 1988, 37, 4225-4233) The reaction was started and incubated for 1 hour at 37 ° C. with shaking, followed by 20% (w / v) trichloroacetic acid, 0.5% (w / v) 2-thio in 0.2 N hydrochloric acid. The reaction was stopped by adding 1 ml of a mixture containing barbituric acid to the above reaction solution, which was boiled at 100 ° C for 30 minutes to develop color. After cooling, the mixture was centrifuged at 3000 rpm for 5 minutes, and the absorbance of the centrifuged supernatant was measured at 532 nm, and the lipid oxidation inhibition rate (%) was expressed by the formula 100-{(SB) / (CB)} × Calculated by calculating 100. Here, C (control) is the absorbance at 532 nm in a methanol solution not containing the test sample, and B (blank) is the absorbance at 532 nm when no rat brain homogenate is added. , S (sample) indicates the absorbance of the test sample, and as a control experiment, the test substance was changed to ginkgo grass powder (negative control), tea leaf powder (positive control) or tea leaf extract (positive control), An experiment similar to the above was performed.

The results are as shown in FIG. Incidentally, it was IC ₅₀ the lipid oxidation inhibiting 50% value. FIG. 1 shows the dilution rate of each test sample that reached IC ₅₀ . Specifically, the final concentration is 0.2 mg / ml for Krachai powder and 0.8 mg equivalent / ml for the extract extract, and this is diluted by 1/2 and the lipid oxidation inhibition rate ( %) And the relationship between the dilution rate and the lipid oxidation inhibition rate (%) was plotted on a graph to determine the dilution rate when the IC ₅₀ was reached. This dilution rate is shown on the vertical axis.

  Krachai powder had a high antioxidant effect, although the antioxidant activity was weaker than that of tea leaves known to have an antioxidant effect. Also, Krachai extract (especially chloroform / methanol extraction, ethanol extraction, methanol extraction) was found to have an antioxidant effect although it was weaker than Krachai powder.

  Therefore, Krachai powder and Krachai extract are considered to be effective in suppressing body oxidation.

(2) Rat brain lipid peroxidation inhibitory experiment using compounds contained in Krachai Next, obtained by the method described in “Preparation of Antioxidant Substance (3) Isolation of 4-hydroxypanduratin” in Krachai The following experiment was conducted using the obtained compound.

This experiment was conducted using the above-mentioned “rat brain lipid peroxidation inhibitory experiment” except that the compound obtained by the method described in “Preparation of Antioxidant Substance (3) Isolation of 4-hydroxypanduratin” was used as a test sample. The experiment was performed using the same method as “1)”. Incidentally, it was IC ₅₀ the lipid oxidation inhibiting 50% value. As a result, IC ₅₀ was found to be as follows.
5-hydroxy-7-methoxyflavanone 230μM
(-)-panduratin A 15μM
5,7-dihydroxyflavanone 210μM
2 ', 6'-dihydroxy4'-methoxychalcone 70μM
2 ', 4'-dihydroxy-6'-methoxychalcone 38μM
(-)-4-hydroxypanduratin A 4.5μM
catechin (control) 17μM
From the above, it was found that the compound having the highest antioxidant activity among the compounds contained in Krachai was (-)-4-hydroxypanduratin A. Furthermore, the antioxidant action of (-)-4-hydroxypanduratin A was about 3.3 times higher than that of (-)-panduratin A.

=== Cerebral nerve cell protective effect experiment ===
In order to examine whether 4-hydroxypanduratin A of the present invention protects brain neurons, the following experiment was conducted.

First, N18-RE-105 cells (mouse neuroblastoma clone N18TG-2 × 18 day old Fisher rat fetal retinal nerve) were mixed with 90% DMEM containing HAT (0.14 mM thymidine, 40 μM aminopterin, 0.1 mM hypoxanthine) and 10%. The cells were seeded in a 25 cm ² culture dish containing a culture solution containing% fetal bovine serum, and cultured in a 37 ° C., 5% CO ₂ incubator. Next, 100 μl of the above culture solution was put in each well of a 96-well microplate, and the above N18-RE-105 cells were put in 20,000 cells / well and cultured for 24 hours. Thereafter, the medium in each well was replaced with a medium containing 10 mM L-glutamic acid and cultured for 48 hours. MTS (tetrazolium) was then added, and the production of formazan crystals (blue) by living cells was quantified by automatic colorimetric analysis. The MTS activity was measured using a kit purchased from Promega (product number: # G5430). The cell death rate (%) was calculated using the formula 100-{(TC) / (BC)} × 100 (%). Here, T (test sample + L-glutamic acid), C (only L-glutamic acid) and B (no addition of test sample and L-glutamic acid) are absorbance at 492 nm. The concentration of test sample required to inhibit 50% of cell death by glutamate was EC _50. The value is shown below.
5-hydroxy-7-methoxyflavanone> 200μM
(-)-panduratin A 13μM
5,7-dihydroxyflavanone> 200μM
2 ', 6'-dihydroxy4'-methoxychalcone 37μM
2 ', 4'-dihydroxy-6'-methoxychalcone 48μM
(-)-4-hydroxypanduratin A 14μM
catechin (control) 160μM
From this, it was found that (-)-4-hydroxypanduratin A has a very high protective effect on brain neurons. It should be noted that (-)-4-hydroxypanduratin A was equivalent to (-)-panduratin A in protecting the brain neurons.

<Example 2: Experiment using processed Pastiers>
=== Preparation of Antioxidant Active Substance ===
(1) Preparation of Paspierre powder 1.5 kg of Paspierre (commercially available product) was lyophilized and powdered with a mixer. Using the same method, tea leaves (commercial product: Jurakuen “Ichikai no Tomo”) and ginkgo grass (commercial product) were also powdered.
(2) Preparation of Paspierre extract Chloroform / methanol extraction (chloroform: methanol = 1: 1), ethanol extraction (using 100% ethanol), methanol extraction (using 100% methanol), hot water extraction (at 100 ° C) Krachai extract was obtained by using hot water. A specific extraction method is as described in Example 1 “Preparation of Krachai extract”. In addition, an extract of tea leaves was also produced using this extraction method.

=== Rat brain lipid peroxidation inhibitory experiment ===
“Paspierre processed product” and “Paspierre extract” are “crachai processed product” and “crachai extract” described in Example 1 “(1) Experiment on inhibition of rat brain lipid peroxidation using processed Krachai”. This experiment was conducted using a method similar to that used for "thing".

The results are as shown in FIG. Incidentally, it was IC ₅₀ the lipid oxidation inhibiting 50% value. FIG. 2 shows the dilution rate of each test sample that reached IC ₅₀ . Specifically, the Paspierre powder or extract is prepared to a final concentration equivalent to 8 mg / ml, diluted in half, and the lipid oxidation inhibition rate (%) is measured for each concentration. The dilution ratio when reaching the IC ₅₀ was determined by plotting the relationship between the percentage of lipid oxidation and the inhibition rate of lipid oxidation (%) in a graph. This dilution rate is shown on the vertical axis.

  The Paspierre powder had about twice as much antioxidant activity as tea leaves known to have antioxidant activity. Further, in the Paspierre extract (chloroform / methanol extraction, ethanol extraction, methanol extraction, hot water extraction), an antioxidant action was observed although it was weaker than Paspierre powder.

  Therefore, it is considered that the Paspierre powder and the Paspierre extract are effective in suppressing the body oxidation.

In one Example of this invention, it is a figure which shows the result of rat brain lipid peroxidation suppression by Krachai, a tea leaf, and a ginkgo. In addition, tea leaves were used as a positive control, and ginkgo grass was used as a negative control. In one Example of this invention, it is a figure which shows the result of the rat brain lipid peroxidation suppression by a Paspierre, a tea leaf, and a ginkgo. In addition, tea leaves were used as a positive control, and ginkgo grass was used as a negative control.

Claims (42)

  An antioxidant composition containing a processed Krachai product.   The antioxidant composition according to claim 1, wherein the processed product is a dried product, a powder, or an extract. A composition that reduces oxidation by active oxygen / radicals,
A composition comprising a processed product of Krachai.   The composition according to claim 3, wherein the processed product is a dried product, a powder, or an extract. A composition for protecting cranial nerve cells for suppressing cranial nerve cell damage caused by excessive intake of glutamic acid,
A composition for protecting cranial nerve cells, comprising a processed product of Krachai.   6. The composition for protecting brain neurons according to claim 5, wherein the processed product is a dried product, a powder, or an extract. A food composition having an antioxidative effect,
A food composition comprising a processed product of Krachai.   The food composition according to claim 7, comprising a health food, a functional food, or a health functional food.   A food containing a processed product of Krachai, having an antioxidant inhibitory effect, and labeled as being used to suppress internal oxidation.   A food containing a processed product of Krachai, having an antioxidant inhibitory effect, and labeled as being used for treatment, prevention, or alleviation of symptoms due to oxidation in the body. A food composition that reduces oxidation by active oxygen / radicals,
A food composition comprising a processed product of Krachai.   The food composition according to claim 11, comprising a health food, a functional food, or a health functional food.   A food containing a processed product of Krachai, characterized by reducing oxidation by active oxygen / radicals and labeled to indicate that it is used to reduce oxidation by active oxygen / radicals present in the body. A food composition for suppressing cranial nerve cell damage due to excessive intake of glutamic acid,
A food composition comprising a processed product of Krachai.   The food composition according to claim 14, comprising a health food, a functional food, or a health functional food.   A food containing a processed product of Krachai, characterized by suppressing cranial nerve cell damage caused by excessive intake of glutamate, and labeled to indicate that it is used for suppressing cranial nerve cell damage caused by excessive intake of glutamate. An indication that it contains Krachai processed products, is characterized by inhibiting neuronal damage caused by excessive intake of glutamate, and is used to treat, prevent, or alleviate symptoms of neurological disorders caused by excessive intake of glutamate Foods marked with   An antioxidant comprising 4-hydroxypanduratin as an active ingredient. An agent that reduces oxidation by active oxygen / radicals,
A drug comprising 4-hydroxypanduratin as an active ingredient. A cranial nerve cell protective agent for suppressing cranial nerve cell damage caused by excessive intake of glutamic acid,
A neuroprotective agent for brain neurons, comprising 4-hydroxypanduratin as an active ingredient. A food composition having an antioxidative effect,
A food composition comprising 4-hydroxypanduratin.   The food composition according to claim 21, which constitutes a health food, a functional food, or a health functional food.   A food that contains 4-hydroxypanduratin and has an anti-oxidation inhibitory effect, and is labeled to indicate that it is used to suppress internal oxidation.   A food that contains 4-hydroxypanduratin and has an antioxidant inhibitory effect, and is labeled as used for treatment, prevention, or alleviation of symptoms due to oxidation in the body. A food composition that reduces oxidation by active oxygen / radicals,
A food composition comprising 4-hydroxypanduratin.   The food composition according to claim 25, which constitutes a health food, a functional food, or a health functional food.   It is characterized by containing 4-hydroxypanduratin and reducing oxidation by active oxygen / radicals, and it is used to reduce oxidation by active oxygen / radicals present in the body. Food. A food composition that suppresses cranial nerve cell damage due to excessive intake of glutamic acid,
A food composition comprising 4-hydroxypanduratin.   The food composition according to claim 28, comprising a health food, a functional food, or a health functional food.   A food containing 4-hydroxypanduratin, characterized in that it inhibits cranial nerve cell damage caused by excessive intake of glutamic acid, and is labeled as used for protecting cranial nerve cells.   An indication that it contains 4-hydroxypanduratin, suppresses cranial nerve cell damage caused by excessive intake of glutamate, and is used for the treatment, prevention, or alleviation of symptoms of neurological diseases caused by excessive intake of glutamate Foods marked with   An antioxidant composition comprising a processed Pastier Pierre.   The antioxidant composition according to claim 32, wherein the processed product is a dried product, a powder, or an extract. A composition that reduces oxidation by active oxygen / radicals,
A composition comprising a processed Paspierre.   The composition according to claim 34, wherein the processed product is a dried product, a powder, or an extract. A food composition having an antioxidative effect,
A food composition comprising a processed pastier product.   37. The food composition according to claim 36, comprising a health food, a functional food, or a health functional food.   A food product containing a processed pastier product and having an antioxidant inhibitory effect, and a label indicating that it is used to suppress internal oxidation.   A food containing a processed Pastier product, having an antioxidant inhibitory effect, and having a label indicating that it is used for the treatment, prevention or alleviation of symptoms due to oxidation in the body. A food composition that reduces oxidation by active oxygen / radicals,
A food composition comprising a processed pastier product.   The food composition according to claim 40, which constitutes a health food, a functional food, or a health functional food. A food product containing a processed Paspierre, characterized by reducing oxidation by active oxygen / radicals, and labeled to indicate that it is used to reduce oxidation by active oxygen / radicals present in the body.

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