JP2009024023A - Lipoxygenase inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide lipoxygenase activity inhibitors that are derived from natural products, have excellent inhibitory action against 5-lipoxygenase and 12-lipoxygenase that participates in the arachidonic acid metabolism, and can prevent and treat allergic diseases, inflammations, further circulatory system diseases and cancer metastasis. <P>SOLUTION: The lipoxygenase inhibitor according to this invention is provided by mixing one or two or more kinds of alcohol extracts of perilla seeds containing luteolin, chrysoeryol, rosmarinic acid, methyl rosmarinate and apigenin. As seeds for the invention, the defatted ones are preferably used. The seeds of Perilla frutescens plant may be used instead of the perilla seeds. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a lipoxygenase inhibitor, and is applied to, for example, pharmaceuticals, quasi-drugs, cosmetics, or food materials that prevent and treat allergic diseases and inflammation.

Arachidonic acid, an essential fatty acid, exists as a component of cell membranes in vivo, and is released from phospholipids by the action of phospholipase when subjected to certain stimuli.
The released arachidonic acid is metabolized by arachidonic acid metabolizing enzymes (lipoxygenase, etc.), prostaglandins (PG) involved in inflammation, thromboxane (TX) involved in thrombus formation, leukotrienes that induce allergies It is converted into substances such as (LT) s and lipoxins (LXs) (see FIG. 1).
Thus, arachidonic acid metabolizing enzymes produce causative substances that are involved in cardiovascular diseases, allergic diseases, inflammation and the like. Therefore, it is considered effective to specifically inhibit arachidonic acid metabolism by enzymes for the prevention and treatment of these diseases.

  Representative enzymes involved in the arachidonic acid metabolic system include 5-lipoxygenase, 12-lipoxygenase, cyclooxygenase and the like. 5-lipoxygenase metabolizes arachidonic acid to 5-hydroxy-6,8,10,14-eicosatetraenoic acid (5-HETE), and induces leukotrienes (LTs) involved in allergic diseases, inflammation, and asthma It is an enzyme. 12-lipoxygenase is an enzyme that metabolizes to 12-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE) involved in arteriosclerosis, allergic diseases, and cancer metastasis using arachidonic acid as a substrate. It is.

Conventionally, as a natural product that inhibits lipoxygenase, for example, caffeic acid and quercetin having a 5-lipoxygenase inhibitory action are known. However, these substances have not yet been put into practical use as enzyme inhibitors.
On the other hand, as substances that inhibit 12-lipoxygenase, few substances showing effective activity are known.
Among other polyphenols, there are natural products having an inhibitory action on lipoxygenase, but all of them are slow in suppressing the extract and cannot be expected to be effective enough to prevent diseases. is there.
In addition, purification of enzyme inhibitors from natural products is difficult to put into practical use due to cost problems.

  The object of the present invention is to have an excellent inhibitory action of 5-lipoxygenase and 12-lipoxygenase involved in arachidonic acid metabolism, and to prevent and treat allergic diseases, inflammation, as well as circulatory system diseases and cancer metastasis. It is to provide a natural product-derived lipoxygenase activity inhibitor to be obtained.

  The lipoxygenase inhibitor of the present invention for solving the above problems comprises an alcohol extract of sesame seeds containing one or more selected from luteolin, chrysoeriol, rosmarinic acid, rosmarinic acid methyl ester and apigenin. It is characterized by.

As the sesame seed, it is desirable to use a defatted sesame seed.
Perilla seeds may be used in place of the sesame seeds.

  The present inventors have conducted various experiments on the extract of sesame seeds, and have discovered a strong inhibitory action of 5-lipoxygenase and 12-lipoxygenase on the ethanol extract of sesame seeds. As a result of earnest research, it was found that luteolin, chrysoeriol, rosmarinic acid, rosmarinic acid methyl ester and apigenin contained in the alcohol extract of sesame seeds have a strong lipoxygenase inhibitory action, and the present invention has been completed. Is. In particular, luteolin exhibits an extremely excellent lipoxygenase inhibitory action that has never been seen before.

Egoma is an annual Lamiaceae oil crop native to East Asia. It resembles a perilla on the whole, with a square stem, round egg-shaped leaves, and white florets in summer. Seeds are slightly larger than perilla and are harvested in autumn.
Oil obtained from sesame seeds is known as sesame oil or camellia oil, and is used as a raw material for food and paint. Oil cake is used as fertilizer and feed.
Conventionally, there are reports of physiologically active substances contained in egoma leaves, but there are few reports on seeds.

As a result of the inventors' investigation, among the lipoxygenase inhibitors (luteolin, chrysoeriol, rosmarinic acid, rosmarinic acid methyl ester and apigenin) contained in the extract of sesame seed, rosmarinic acid and rosmarinic acid methyl ester have already been obtained. JP-A-1-121217 discloses that 5-lipoxygenase inhibitory action of rosmarinic acid or its derivatives extracted from fresh leaves of Lamiaceae plants is disclosed.
As for apigenin, the 5-lipoxygenase inhibitory action is described in JP-A-8-510735.
However, among these substances, the lipoxygenase inhibitory action of luteolin, which has the largest inhibitory action, has not been known so far, and the lipoxygenase inhibitory action of chrysoeriol has also been clarified by this study. .

  In addition, the present inventors also conducted various experiments on perilla seeds, which are the same Lamiaceae seeds as sesame seeds. And rosmarinic acid methyl ester and apigenin).

ルテオリンは、一般にマメ科の植物（ジギタリス等）中に配糖体として存在することが知られる。ルテオリンの生理活性については、抗酸化活性、ヒアルロニダーゼ阻害活性等が知られている。
本発明において、ルテオリンをリポキシゲナーゼ阻害剤の有効成分として使用する場合には、エゴマおよびシソの種子および葉から抽出・精製する他、柑橘系の果皮等からも抽出・精製することが可能である。ただし、エゴマおよびシソの葉に含まれるルテオリンは微量であり、工業化に際しては、これらの種子を用いるのが望ましい。 The structural formula of luteolin is as shown below.

Luteolin is generally known to exist as a glycoside in leguminous plants (such as digitalis). As for the physiological activity of luteolin, antioxidant activity, hyaluronidase inhibitory activity and the like are known.
In the present invention, when luteolin is used as an active ingredient of a lipoxygenase inhibitor, it can be extracted and purified from sesame seeds and leaves as well as from citrus peels and the like. However, the amount of luteolin contained in the leaves of perilla and perilla is very small, and it is desirable to use these seeds for industrialization.

クリソエリオールの生理活性については、抗癌性、抗菌性を有するとの報告がわずかにある程度である。
本発明において、クリソエリオールをリポキシゲナーゼ阻害剤の有効成分として使用する場合、エゴマまたはシソの種子および葉から抽出・精製することが可能である。 The structural formula of chrysoeriol is as shown below.

Regarding the physiological activity of chrysoeriol, there are only a few reports that it has anticancer and antibacterial properties.
In the present invention, when chrysoeriol is used as an active ingredient of a lipoxygenase inhibitor, it can be extracted and purified from seeds and leaves of egoma or perilla.

It is desirable to use ethanol as a solvent for extracting luteolin and chrysoeriol using egoma seeds and perilla seeds as raw materials. This is because when ethanol is used, the active ingredient can be efficiently extracted and used at the same time for both external and edible uses. Moreover, when using as an external preparation, it is also possible to use ethyl acetate, acetone, methanol, butanol, etc. other than ethanol.
The alcohol concentration is preferably adjusted to 70 to 85% (v / v). If it is less than 70% (v / v), the extraction amount of the active ingredient becomes insufficient, and if it exceeds 85% (v / v), the oil content of the sesame seeds easily dissolves in the alcohol. is there.
In addition, in order to improve the content rate of an active ingredient, it is desirable to perform alcohol extraction repeatedly at various concentrations.

  Alcohol extracts of sesame seeds and perilla seeds usually contain all of luteolin, chrysoeriol, rosmarinic acid, rosmarinic acid methyl ester and apigenin as lipoxygenase inhibitors. However, even if any substance does not exist depending on the extraction conditions or the like, it is sufficient that at least one or more of these substances are included.

  The reason why alcoholic extracts of egoma seeds or perilla seeds are distributed with ethyl acetate and water is that the concentration of the active ingredient can be significantly increased by these distributions. Among the distribution layers, the ethyl acetate layer contains luteolin, chrysoeriol, rosmarinic acid, rosmarinic acid methyl ester and apigenin at high concentrations, and inactive components such as glycosides migrate to the aqueous layer. Therefore, by separating the ethyl acetate layer, the active ingredient can be efficiently concentrated.

The alcohol extract and ethyl acetate partition can be incorporated into a lipoxygenase inhibitor as they are, but the active ingredients contained in these can be purified.
For example, when purifying luteolin, the ethyl acetate layer was subjected to silica gel column chromatography (chloroform: methanol = 10: 1), and the most active fraction was recovered, and the mixed solvent (chloroform: methanol = 15: 1) Luteolin can be isolated from the insoluble fraction.

In the present invention, when using sesame seeds or perilla seeds, the concentration of the active ingredient can be greatly increased by alcohol extraction using these defatted products. That is, when oils of sesame seeds and perilla seeds are extracted with an organic solvent, the oils hardly contain physiologically active components such as luteolin and chrysoeriol, and the active ingredients are concentrated in the defatted koji.
As an organic solvent for degreasing, for example, hexane may be used. This is because the extracted oil can be used as an edible oil and the extract from the defatted egoma can be used as a food material. Moreover, when using the extract from defatted egoma as an external preparation (mainly uses other than food), it is also possible to use not only hexane but another nonpolar solvent.

Examples of the use of the lipoxygenase inhibitor of the present invention include pharmaceuticals, quasi drugs, cosmetics, food materials and the like.
In particular, when the lipoxygenase inhibitor of the present invention is used as an antiallergic agent or antiinflammatory agent, it is possible to obtain excellent antiallergic and antiinflammatory effects resulting from inhibition of lipoxygenase activity.

The administration method of the lipoxygenase inhibitor of the present invention may be either oral or parenteral administration. For oral administration, it can be administered as soft / hard capsules or tablets, granules, fine granules, powders, and for parenteral administration, injections, drops and solid or suspension viscous In addition to administration in a dosage form such as a suppository so that sustained mucosal absorption can be maintained as a liquid, administration into a local tissue, intradermal, subcutaneous, intramuscular and intravenous injection, topical application, spraying, suppository, External administration methods such as intravesical injection can also be used.
The dosage may vary depending on the administration method and the malignancy of the disease, the age of the patient, the medical condition and general condition, the progression of the disease, etc. In adults, it is usually 0.5 to 5000 mg as an active ingredient per day. Usually 0.5 to 3000 mg is appropriate.
The active ingredient concentration of the lipoxygenase inhibitor can be appropriately changed depending on the dosage form, but is usually about 0.3 to 15.0 w% when administered orally or by mucosal absorption, when administered parenterally Is preferably about 0.01 to 10 w%. The above dose is an example, and can be appropriately changed according to various situations.

When the lipoxygenase inhibitor of the present invention is used as a pharmaceutical, a quasi-drug, or a cosmetic, it can be used in combination with various components generally used for pharmaceuticals.
For example, oil (animal and vegetable oil, mineral oil, ester oil, wax oil, silicone oil, higher alcohol, phospholipids, fatty acids, etc.), surfactant (anionic, cationic, amphoteric or nonionic surfactant) , Vitamins (vitamin A group, vitamin B group, folic acid, nicotinic acid, pantothenic acid, biotins, vitamin C group, vitamin D group, vitamin E group, other ferulic acid, γ-oryzanol, etc.), UV absorber ( p-aminobenzoic acid, anthranyl, salicylic acid, coumarin, benzotriazole, tetrazole, imidazoline, pyrimidine, dioxane, furan, pyrone, camphor, nucleic acid, allantoin and their derivatives, amino acid compounds, shikonin, baicalin, baicalein, berberine, etc. ), Antioxidants (stearic acid ester, Dihydrogua cetelenic acid, dibutylhydroxytoluene, butylhydroxyanisole, parahydroxyanisole, propyl gallate, sesamol, sesamolin, gossypol, etc.), thickener (hydroxyethyl cellulose, ethyl cellulose, carboxyethyl cellulose, methyl cellulose, carboxymethyl cell) Roll, sodium carboxymethyl cellulose, hydroxypropyl cellulose, nitrocellulose, polyvinyl alcohol, polyvinyl methyl ether, polyvinyl pyrrolidone, polyvinyl methacrylate, polyacrylate, carboxyvinyl polymer, gum arabic, tragacanth gum, agar, casein, dextrin, gelatin, Pectin, starch, alginic acid and its salts), moisturizer (propylene glycol, 1 , 3-butylene glycol, polyethylene glycol, glycerin, chondroitin sulfate and salts thereof, hyaluronic acid and salts thereof, sodium lactate, etc.), other lower alcohols, polyhydric alcohols, water-soluble polymers, pH adjusters, antiseptic and antibacterial agents Agent, coloring agent, fragrance, refreshing agent, stabilizer, animal / plant extract, animal / plant protein and its degradation product, animal / plant polysaccharide and its degradation product, animal / plant glycoprotein and its degradation Compound with microbial culture metabolic component, blood flow promoting agent, anti-inflammatory agent, cell activator, amino acid and its salt, keratolytic agent, astringent agent, wound healing agent, foam increasing agent, oral agent, deodorant / deodorant Can be used.

  In addition, the form in the case of using as these pharmaceuticals, quasi-drugs, or cosmetics can be selected arbitrarily, for example, cream, ointment, lotion, lotion, emulsion, pack, oil, soap (medicine soap is also used) Including), facial cleanser, bath preparation, shampoo, rinse, spray and the like. In addition, it can be used as a composition for oral cavity caused by stomatitis or the like on a nonwoven fabric such as sanitary cotton or wet tissue.

  In addition, when the lipoxygenase inhibitor of the present invention is used as a food material, it can be applied to general foods such as confectionery, noodles, soups, beverages, health foods, dietary supplements and the like. For example, a lipoxygenase inhibitor spray-dried or freeze-dried with powdered cellulose can be easily mixed into food by making powder, granules, tablets or solutions.

5-HETE, which is a metabolite of 5-lipoxygenase, generates leukotrienes and induces allergies and inflammatory reactions, and 12-HETE, an arachidonic acid metabolite of 12-lipoxygenase, is involved in arteriosclerosis and cancer metastasis. It is said to be a physiologically active substance. Therefore, according to the present invention, allergic diseases, inflammation, cardiovascular diseases, cancer, and the like can be effectively prevented and treated by selectively suppressing their production.
Moreover, according to the present invention, an excellent lipoxygenase inhibitor can be extracted from natural products such as sesame seeds, and can be effectively used as a food material for suppressing allergies and inflammation associated therewith.
Furthermore, according to the present invention, a lipoxygenase inhibitor can be obtained from defatted sesame after recovery of edible oil from sesame seeds, thereby providing a new use of defatted sesame.

Examples of the present invention will be described below.
[Preparation of Lipoxygenase Inhibitor]
As shown in FIG. 2, first, crushed sesame seeds were refluxed with hexane, and then the residue (defatted sesame seeds) was refluxed with 80% (v / v) ethanol.
Next, the ethanol extract obtained by refluxing 80% (v / v) ethanol was distributed between hexane and 80% (v / v) methanol. After the solvent was distilled off from the methanol layer, it was further partitioned between ethyl acetate and water. . After separating the ethyl acetate layer and the aqueous layer, the solvent of the ethyl acetate layer was distilled off to obtain an ethyl acetate partition.

Subsequently, this ethyl acetate distribution was subjected to silica gel column chromatography (chloroform: methanol = 10: 1) and fractionated into fractions 1 to 3.
Among these, fraction 1 eluted first was suspended in a mixed solvent (chloroform: methanol = 20: 1), and Cresoeriol was isolated from the insoluble fraction.
Further, fraction 2 to be eluted next was suspended in a mixed solvent (chloroform: methanol = 15: 1), and luteolin was isolated from the insoluble fraction.

The purified luteolin product isolated from the sesame seeds was designated as Example 1, and the purified chrysoeriol product was designated as Example 2.
Moreover, the ethanol extract obtained by distilling off the ethanol extract shown in FIG. 2 was used as Example 3, and the ethyl acetate distribution obtained by distilling off the ethyl acetate layer obtained by partitioning the ethanol extract was carried out. Example 4 was adopted.

[5-lipoxygenase inhibition test]
Next, as to the lipoxygenase inhibitors of Examples 1 to 4, 5-lipoxygenase inhibition tests were performed as described below.
(1) Preparation of 5-lipoxygenase enzyme solution
5% (w / v) glycogen (20 ml / kg) was intraperitoneally administered to Wister-King normal rats (150 to 400 g), and the intraperitoneal fluid was collected. A suspension of polymorphonuclear leukocytes obtained by centrifugation in 50 mM phosphate buffer (pH 7.4) was homogenized, and the supernatant was used as an enzyme solution for 5-lipoxygenase.

(2) Measurement of 5-lipoxygenase activity inhibition First, samples of different concentrations (4 to 6 types) were prepared for Examples 1 to 4, and 0.02 ml of enzyme solution (2 mg protein / ml) and [1 - the ¹⁴ C] arachidonic acid (0.05 [mu] Ci), each sample (0.02 ml) and Ca ^2+, the presence of ATP, and incubated for 5 minutes at 37 ° C..
The reaction was then stopped with 0.2 ml of 0.5N formic acid and the metabolites were extracted with 3 ml of ethyl acetate.
After this extract was subjected to TLC at 4 ° C. (petroleum ether: ether: acetic acid = 50: 50: 1 (v / v)), metabolites were quantified by autoradiography to determine 5-lipoxygenase inhibitory activity. . The 5-lipoxygenase inhibitory activity was determined using the radioactivity obtained in the absence of the sample as a control.

  As a comparative example, rosmarinic acid (Comparative Example 1) and rosmarinic acid methyl ester (Comparative Example 2) obtained by purifying fraction 3 shown in FIG. The 5-lipoxygenase inhibitory activity was also measured in the same manner for the partition (Comparative Example 3) and the water partition obtained by distilling off the ethyl acetate / water partition aqueous layer (Comparative Example 4). The results are shown in FIG. 3 to FIG. 5, FIG. 7 and FIG.

The results showed excellent 5-lipoxygenase inhibitory activity in both Examples 1 to 4. In particular, Example 1 (Luteolin) shown in FIG. 3 has stronger inhibitory activity than Comparative Example 1 (rosmarinic acid) and Comparative Example 2 (rosmarinic acid methyl ester), which have been reported to have a 5-lipoxygenase inhibitory action. It was a thing.
In addition, as shown in FIG. 5, Example 4 (ethyl acetate partition) showed an inhibitory activity superior to that of Example 3 (ethanol extract), which is less water-soluble than ethanol extract. This is probably because the active ingredient is concentrated in the ethyl acetate layer as a result of removing the water-soluble and oil-soluble substances. The inhibitory activity of Example 3 (ethanol extract) and Example 4 (ethyl acetate partition) seems to correlate with the content of luteolin.

表１に示すように、実施例１（ルテオリン）のＩＣ５０は、０．１μＭであり、他のものに比べきわめて高い、優れた５−リポキシゲナーゼ阻害活性を示す結果となった。 Here, for Example 1 (Luteolin), Example 2 (Chrysoeriol), Comparative Example 1 (Rosmarinic Acid) and Comparative Example 2 (Rosmarinic Acid Methyl Ester), the IC50 (50% inhibitory concentration) of 5-lipoxygenase was calculated. Table 1 shows the obtained results.
In addition, since IC50 was calculated | required by the same method also about the comparative example 5 (caffeic acid) and the comparative example 6 (quercetin), it shows in Table 1 collectively.

As shown in Table 1, the IC50 of Example 1 (Luteolin) was 0.1 μM, which was an extremely high result compared to the others, showing excellent 5-lipoxygenase inhibitory activity.

[Inhibition test of 12-lipoxygenase]
Next, for Examples 1 to 4, a 12-lipoxygenase inhibition test was performed as described below.
(1) Preparation of 12-lipoxygenase enzyme solution
Blood was collected from Wister-King normal rats (150 to 400 g), and 0.5 mM EDTA was added as an anticoagulant. The centrifuged supernatant (1200 rpm, 10 minutes) was further centrifuged (3000 rpm, 10 minutes), and the precipitated platelets were added to 25 mM Tris-HCl buffer (pH 7.4) / 1 mM EDTA. Washed with / 130 mM NaCl. The obtained platelet 1 mM EDTA suspension homogenate was used as a 12-lipoxygenase enzyme solution.

(2) Measurement of activity inhibition of 12-lipoxygenase For Examples 1 to 4, samples (4 to 6 types) having different concentrations were prepared, and 0.13 ml of enzyme solution (2 mg protein / ml) and [1- ¹⁴ C] arachidonic acid (0.05 μCi) was incubated at 37 ° C. for 5 minutes in the presence of the sample (0.02 ml).
The reaction was then stopped with 0.2 ml of 0.5N formic acid and the metabolites were extracted with 3 ml of ethyl acetate.
After subjecting this extract to TLC at room temperature (chloroform: methanol: acetic acid: water = 90: 8: 1: 0.8 (v / v)), the metabolite was quantified by autoradiography to inhibit 12-lipoxygenase. Activity was sought. In addition, 12-lipoxygenase inhibitory activity was calculated | required as a control the radioactivity obtained in the absence of a lipoxygenase inhibitor.

  As comparative examples, rosmarinic acid (Comparative Example 1) and rosmarinic acid methyl ester (Comparative Example 2) obtained by purifying fraction 3 shown in FIG. The 5-lipoxygenase inhibitory activity was also measured in the same manner for the resulting hexane partition (Comparative Example 3) and the water partition obtained by distilling off the ethyl acetate / water partition aqueous layer (Comparative Example 4). did. The results are shown in FIG. 3, FIG. 4 and FIGS.

The results showed excellent 12-lipoxygenase inhibitory activity in both Examples 1 to 4. In particular, Example 1 (Luteolin) had a very strong inhibitory activity, similar to the inhibitory activity of 5-lipoxygenase.
Moreover, as shown in FIG. 6, Example 4 (ethyl acetate partition) is stronger than the ethanol extract (Example 3) because the active ingredient in the ethanol extract is concentrated in the ethyl acetate partition. Inhibitory activity was shown.

[Inflammation suppression test for TPA-induced ear edema in mice]
Next, Example 1, Example 3, and Example 4 were tested for inflammation-suppressing action against TPA-induced ear edema.
(1) Transdermal administration A sample (Example 1, Example 3, Example 4) prepared in a predetermined amount was mixed with an acetone solution of TPA (0.8 μg / 20 μl / ear) in a mouse (ICR system, 5 weeks old, Applied to the right auricle of (Claire). Four hours after administration, the weights of the right and left auricle pieces were measured, the swelling rate of the right auricle piece relative to the left auricle piece was calculated, and the swelling inhibition rate for the control group was further obtained. In addition, a control is a test conducted under the same conditions for a group not administered with a sample.
As a comparative example, data reported for NDGA (Comparative Example 7) is shown in Table 2.

  As shown in Table 2, Comparative Example 7 (NDGA), which is a 5-lipoxygenase inhibitor, has a 40% inhibition rate at 0.5 mg / ear, whereas Example 1 (Luteolin) has 0.3 mg / ear. 100%, Example 3 (ethanol extract) was 25% at 1 mg / ear, and Example 4 (ethyl acetate partition) was 45% at 0.3 mg / ear, which suppressed ear edema due to TPA. That is, it can be seen that Example 1, Example 3 and Example 4 all show excellent swelling suppression rates. Moreover, about Example 1, the anti-inflammatory action correlated with the density | concentration of luteolin was recognized.

(2) Oral administration As a sample, Example 4 (ethanol extract) was mixed with 1% food, and after 5 g per day for 4 weeks, a TPA acetone solution (0.8 μg / 20 μl / ear) was added to a mouse (ICR system). 5 weeks old, Claire) was applied to the right auricle. Four hours after administration, the weights of the right and left auricle pieces were measured, the swelling rate of the right auricle piece relative to the left auricle piece was calculated, and the swelling inhibition rate for the control group was further obtained. In addition, a control is a test conducted under the same conditions for a group not administered with a sample. The results are shown in Table 2.

  As shown in Table 2, the swelling suppression rate due to 1% bait mixing of Example 4 (ethanol extract) was 20%, and an excellent anti-inflammatory effect was observed even in oral administration. In addition, the increase and decrease of the body weight were not recognized compared with the control group.

[Allergy suppression test for oxazolone-induced ear edema in mice]
Next, Example 1 and Example 3 were tested for anti-inflammatory action against oxazolone-induced ear edema.
(1) Transdermal administration Under anesthesia with Nembutal, the abdomen of a mouse (ICR system, 5 weeks old, Claire) was shaved with a clipper and an oxazolone ethanol solution (500 μg / 100 μl) was applied (sensitization). Five days after sensitization, an oxazolone acetone solution (100 μg / 20 μl) added with a predetermined amount of sample (Example 1) was applied to the right auricle (challenge), and the weight of the left and right auricle pieces 48 hours later was measured. Using the weight ratio of the right auricle piece to the left auricle piece as the swelling rate, the swelling suppression rate for the control group was determined. The control is a test conducted under the same conditions for the group not administered with the sample.
As comparative examples, reported data for NDGA (Comparative Example 7) ketoprofen (Comparative Example 8), phenidone (Comparative Example 9), and mepyramine (Comparative Example 10) are shown in Table 3.

As shown in Table 3, Example 1 showed a high swelling suppression rate with a smaller dose compared to Comparative Examples 7-10. That is, it can be seen that luteolin, which is an active ingredient, exhibits an excellent antiallergic action.
(2) Oral administration During the period from the start of administration to pinna weight measurement, the sample (Example 3) was mixed in 1% food and given 5 g per day. Eight days after the start of administration, the shaved abdomen was sensitized with an oxazolone ethanol solution, and five days later, an oxazolone acetone solution was applied to the right auricle (challenge), and the weight of the left and right auricle pieces was measured 48 hours later. . Using the weight ratio of the right auricle piece to the left auricle piece as the swelling rate, the swelling suppression rate for the control group was determined. The control is a test conducted under the same conditions for the group not administered with the sample. The results are shown in Table 3.

  As shown in Table 3, Example 3 inhibited oxazolone-induced ear edema by 34% with 1% bait mixing. That is, an antiallergic action was observed even by oral administration. In addition, about the mouse | mouth which administered Example 3, the increase / decrease in body weight was not recognized compared with the control group.

It is action explanatory drawing for demonstrating the physiological activity of an arachidonic acid metabolic enzyme. FIG. 3 is a process diagram for explaining a method for preparing a lipoxygenase inhibitor according to an example of the present invention. It is a characteristic view which shows the relationship between the density | concentration of the luteolin purified material by the Example of this invention, and 5-lipoxygenase and 12-lipoxygenase inhibitory activity. It is a characteristic view which shows the relationship between the density | concentration of the chrysoeriol purified product by the Example of this invention, and 5-lipoxygenase and 12-lipoxygenase inhibitory activity. It is a characteristic view which shows the relationship between the density | concentration of the ethanol extract by the Example of this invention, and an ethyl acetate partition, and 5-lipoxygenase inhibitory activity. It is a characteristic view which shows the relationship between the density | concentration of the ethanol extract by the Example of this invention, and an ethyl acetate partition, and 12-lipoxygenase inhibitory activity. It is a characteristic view which shows the relationship between the density | concentration of the rosmarinic acid refinement | purification material by the comparative example of this invention, and 5-lipoxygenase and 12-lipoxygenase inhibitory activity. It is a characteristic view which shows the relationship between the density | concentration of the rosmarinic acid methyl ester refinement | purification material by the comparative example of this invention, and 5-lipoxygenase and 12-lipoxygenase inhibitory activity.

Claims (5)

  A lipoxygenase inhibitor comprising an alcohol extract of sesame seeds containing one or more selected from luteolin, chrysoeriol, rosmarinic acid, rosmarinic acid methyl ester and apigenin.   The lipoxygenase inhibitor according to claim 1, wherein a defatted sesame seed is used as the sesame seed.   The lipoxygenase inhibitor according to claim 1 or 2, wherein perilla seeds are used instead of the sesame seeds.   The antiallergic agent containing the lipoxygenase inhibitor of any one of the said Claims 1-3.   The anti-inflammatory agent containing the lipoxygenase inhibitor of any one of the said Claims 1-3.

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