JP2005132791A - Antiallergic agent - Google Patents
Antiallergic agent Download PDFInfo
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- JP2005132791A JP2005132791A JP2003372430A JP2003372430A JP2005132791A JP 2005132791 A JP2005132791 A JP 2005132791A JP 2003372430 A JP2003372430 A JP 2003372430A JP 2003372430 A JP2003372430 A JP 2003372430A JP 2005132791 A JP2005132791 A JP 2005132791A
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- hesperidin
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- 239000000043 antiallergic agent Substances 0.000 title claims abstract description 36
- ARGKVCXINMKCAZ-UHFFFAOYSA-N neohesperidine Natural products C1=C(O)C(OC)=CC=C1C1OC2=CC(OC3C(C(O)C(O)C(CO)O3)OC3C(C(O)C(O)C(C)O3)O)=CC(O)=C2C(=O)C1 ARGKVCXINMKCAZ-UHFFFAOYSA-N 0.000 claims abstract description 63
- 239000001100 (2S)-5,7-dihydroxy-2-(3-hydroxy-4-methoxyphenyl)chroman-4-one Substances 0.000 claims abstract description 62
- QUQPHWDTPGMPEX-UHFFFAOYSA-N Hesperidine Natural products C1=C(O)C(OC)=CC=C1C1OC2=CC(OC3C(C(O)C(O)C(COC4C(C(O)C(O)C(C)O4)O)O3)O)=CC(O)=C2C(=O)C1 QUQPHWDTPGMPEX-UHFFFAOYSA-N 0.000 claims abstract description 62
- QUQPHWDTPGMPEX-UTWYECKDSA-N aurantiamarin Natural products COc1ccc(cc1O)[C@H]1CC(=O)c2c(O)cc(O[C@@H]3O[C@H](CO[C@@H]4O[C@@H](C)[C@H](O)[C@@H](O)[C@H]4O)[C@@H](O)[C@H](O)[C@H]3O)cc2O1 QUQPHWDTPGMPEX-UTWYECKDSA-N 0.000 claims abstract description 62
- APSNPMVGBGZYAJ-GLOOOPAXSA-N clematine Natural products COc1cc(ccc1O)[C@@H]2CC(=O)c3c(O)cc(O[C@@H]4O[C@H](CO[C@H]5O[C@@H](C)[C@H](O)[C@@H](O)[C@H]5O)[C@@H](O)[C@H](O)[C@H]4O)cc3O2 APSNPMVGBGZYAJ-GLOOOPAXSA-N 0.000 claims abstract description 62
- VUYDGVRIQRPHFX-UHFFFAOYSA-N hesperidin Natural products COc1cc(ccc1O)C2CC(=O)c3c(O)cc(OC4OC(COC5OC(O)C(O)C(O)C5O)C(O)C(O)C4O)cc3O2 VUYDGVRIQRPHFX-UHFFFAOYSA-N 0.000 claims abstract description 62
- QUQPHWDTPGMPEX-QJBIFVCTSA-N hesperidin Chemical compound C1=C(O)C(OC)=CC=C1[C@H]1OC2=CC(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO[C@H]4[C@@H]([C@H](O)[C@@H](O)[C@H](C)O4)O)O3)O)=CC(O)=C2C(=O)C1 QUQPHWDTPGMPEX-QJBIFVCTSA-N 0.000 claims abstract description 62
- 229940025878 hesperidin Drugs 0.000 claims abstract description 62
- 235000020971 citrus fruits Nutrition 0.000 claims abstract description 43
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 108
- 239000000284 extract Substances 0.000 claims description 50
- 241000555678 Citrus unshiu Species 0.000 claims description 35
- 239000000843 powder Substances 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 10
- 238000000605 extraction Methods 0.000 claims description 6
- 230000005070 ripening Effects 0.000 claims 2
- 230000003266 anti-allergic effect Effects 0.000 abstract description 25
- 208000026935 allergic disease Diseases 0.000 abstract description 8
- 239000000469 ethanolic extract Substances 0.000 abstract description 7
- 208000035285 Allergic Seasonal Rhinitis Diseases 0.000 abstract description 6
- 206010012438 Dermatitis atopic Diseases 0.000 abstract description 6
- 208000006673 asthma Diseases 0.000 abstract description 6
- 201000008937 atopic dermatitis Diseases 0.000 abstract description 6
- 208000030603 inherited susceptibility to asthma Diseases 0.000 abstract description 6
- 206010039085 Rhinitis allergic Diseases 0.000 abstract description 3
- 201000010105 allergic rhinitis Diseases 0.000 abstract description 3
- 230000006872 improvement Effects 0.000 abstract description 2
- 238000011282 treatment Methods 0.000 abstract description 2
- 206010048908 Seasonal allergy Diseases 0.000 abstract 1
- 238000011321 prophylaxis Methods 0.000 abstract 1
- 235000013399 edible fruits Nutrition 0.000 description 29
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 26
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 18
- 206010030113 Oedema Diseases 0.000 description 16
- 206010040914 Skin reaction Diseases 0.000 description 14
- 230000035483 skin reaction Effects 0.000 description 14
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- 239000000243 solution Substances 0.000 description 14
- LOTKRQAVGJMPNV-UHFFFAOYSA-N 1-fluoro-2,4-dinitrobenzene Chemical compound [O-][N+](=O)C1=CC=C(F)C([N+]([O-])=O)=C1 LOTKRQAVGJMPNV-UHFFFAOYSA-N 0.000 description 10
- 239000004480 active ingredient Substances 0.000 description 8
- 230000000694 effects Effects 0.000 description 8
- 241000207199 Citrus Species 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- 244000183685 Citrus aurantium Species 0.000 description 5
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- 230000002401 inhibitory effect Effects 0.000 description 5
- NTYJJOPFIAHURM-UHFFFAOYSA-N Histamine Chemical compound NCCC1=CN=CN1 NTYJJOPFIAHURM-UHFFFAOYSA-N 0.000 description 4
- 108010058846 Ovalbumin Proteins 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- CGIGDMFJXJATDK-UHFFFAOYSA-N indomethacin Chemical compound CC1=C(CC(O)=O)C2=CC(OC)=CC=C2N1C(=O)C1=CC=C(Cl)C=C1 CGIGDMFJXJATDK-UHFFFAOYSA-N 0.000 description 4
- 230000006698 induction Effects 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 229940092253 ovalbumin Drugs 0.000 description 4
- XMDUIJHKDOLYSR-UHFFFAOYSA-M sodium;2,3-dinitrobenzenesulfonate Chemical compound [Na+].[O-][N+](=O)C1=CC=CC(S([O-])(=O)=O)=C1[N+]([O-])=O XMDUIJHKDOLYSR-UHFFFAOYSA-M 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 230000009285 allergic inflammation Effects 0.000 description 3
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- 238000010998 test method Methods 0.000 description 3
- 206010002198 Anaphylactic reaction Diseases 0.000 description 2
- 235000007716 Citrus aurantium Nutrition 0.000 description 2
- 241001672694 Citrus reticulata Species 0.000 description 2
- 206010020751 Hypersensitivity Diseases 0.000 description 2
- 240000007594 Oryza sativa Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 241001093501 Rutaceae Species 0.000 description 2
- 230000007815 allergy Effects 0.000 description 2
- VXAUWWUXCIMFIM-UHFFFAOYSA-M aluminum;oxygen(2-);hydroxide Chemical compound [OH-].[O-2].[Al+3] VXAUWWUXCIMFIM-UHFFFAOYSA-M 0.000 description 2
- 208000003455 anaphylaxis Diseases 0.000 description 2
- 230000003110 anti-inflammatory effect Effects 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
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- 210000005069 ears Anatomy 0.000 description 2
- 230000003203 everyday effect Effects 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 229960001340 histamine Drugs 0.000 description 2
- 210000003630 histaminocyte Anatomy 0.000 description 2
- 239000010903 husk Substances 0.000 description 2
- 229960000905 indomethacin Drugs 0.000 description 2
- 230000000977 initiatory effect Effects 0.000 description 2
- XMGQYMWWDOXHJM-UHFFFAOYSA-N limonene Chemical compound CC(=C)C1CCC(C)=CC1 XMGQYMWWDOXHJM-UHFFFAOYSA-N 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- YRCWQPVGYLYSOX-UHFFFAOYSA-N synephrine Chemical compound CNCC(O)C1=CC=C(O)C=C1 YRCWQPVGYLYSOX-UHFFFAOYSA-N 0.000 description 2
- HJRJRUMKQCMYDL-UHFFFAOYSA-N 1-chloro-2,4,6-trinitrobenzene Chemical compound [O-][N+](=O)C1=CC([N+]([O-])=O)=C(Cl)C([N+]([O-])=O)=C1 HJRJRUMKQCMYDL-UHFFFAOYSA-N 0.000 description 1
- SSXJHQZOHUYEGD-UHFFFAOYSA-N 3-Methoxynobiletin Chemical compound C1=C(OC)C(OC)=CC=C1C1=C(OC)C(=O)C2=C(OC)C(OC)=C(OC)C(OC)=C2O1 SSXJHQZOHUYEGD-UHFFFAOYSA-N 0.000 description 1
- 239000001606 7-[(2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxyoxan-2-yl]oxy-5-hydroxy-2-(4-hydroxyphenyl)chroman-4-one Substances 0.000 description 1
- 244000223760 Cinnamomum zeylanicum Species 0.000 description 1
- 235000000228 Citrus myrtifolia Nutrition 0.000 description 1
- 241001561395 Citrus natsudaidai Species 0.000 description 1
- 235000016646 Citrus taiwanica Nutrition 0.000 description 1
- 206010012442 Dermatitis contact Diseases 0.000 description 1
- 235000011511 Diospyros Nutrition 0.000 description 1
- 244000236655 Diospyros kaki Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 235000017317 Fortunella Nutrition 0.000 description 1
- OBIOZWXPDBWYHB-UHFFFAOYSA-N Nobiletin Natural products C1=CC(OC)=CC=C1C1=C(OC)C(=O)C2=C(OC)C(OC)=C(OC)C(OC)=C2O1 OBIOZWXPDBWYHB-UHFFFAOYSA-N 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 239000003470 adrenal cortex hormone Substances 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- 239000002260 anti-inflammatory agent Substances 0.000 description 1
- 229940121363 anti-inflammatory agent Drugs 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 235000021028 berry Nutrition 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 235000017803 cinnamon Nutrition 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 229920002055 compound 48/80 Polymers 0.000 description 1
- 208000010247 contact dermatitis Diseases 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
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- 239000007884 disintegrant Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 229930182486 flavonoid glycoside Natural products 0.000 description 1
- 150000007955 flavonoid glycosides Chemical class 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 230000005094 fruit set Effects 0.000 description 1
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- 238000010438 heat treatment Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 229940087305 limonene Drugs 0.000 description 1
- 235000001510 limonene Nutrition 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000401 methanolic extract Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- DFPMSGMNTNDNHN-ZPHOTFPESA-N naringin Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](OC=2C=C3O[C@@H](CC(=O)C3=C(O)C=2)C=2C=CC(O)=CC=2)O[C@H](CO)[C@@H](O)[C@@H]1O DFPMSGMNTNDNHN-ZPHOTFPESA-N 0.000 description 1
- 229930019673 naringin Natural products 0.000 description 1
- 229940052490 naringin Drugs 0.000 description 1
- ARGKVCXINMKCAZ-UZRWAPQLSA-N neohesperidin Chemical compound C1=C(O)C(OC)=CC=C1[C@H]1OC2=CC(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O[C@H]3[C@@H]([C@H](O)[C@@H](O)[C@H](C)O3)O)=CC(O)=C2C(=O)C1 ARGKVCXINMKCAZ-UZRWAPQLSA-N 0.000 description 1
- MRIAQLRQZPPODS-UHFFFAOYSA-N nobiletin Chemical compound C1=C(OC)C(OC)=CC=C1C1=CC(=O)C2=C(OC)C(OC)=C(OC)C(OC)=C2O1 MRIAQLRQZPPODS-UHFFFAOYSA-N 0.000 description 1
- 239000000041 non-steroidal anti-inflammatory agent Substances 0.000 description 1
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- 238000003809 water extraction Methods 0.000 description 1
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
Description
本発明は、ヘスペリジンを含有する未成熟な柑橘類果実のエタノール抽出エキスあるいは乾燥粉末を有効成分として含有することを特徴とする抗アレルギー剤に関する。 The present invention relates to an antiallergic agent characterized by containing an ethanol extract of immature citrus fruits containing hesperidin or a dry powder as an active ingredient.
従来、抗アレルギー剤および抗炎症剤としては、グルココルチコイド型の副腎皮質ホルモン剤や、インドメタシンなどの非ステロイド性抗炎症剤などが用いられている。しかしながら、ステロイドホルモン剤は抗アレルギー作用および抗炎症作用を有するものの副作用が強いという問題があり、また、インドメタシンなどは抗炎症作用のみを有するため抗アレルギー剤としては有用でなかった。 Conventionally, as antiallergic agents and anti-inflammatory agents, glucocorticoid-type corticosteroid agents, non-steroidal anti-inflammatory agents such as indomethacin, and the like have been used. However, steroid hormone agents have anti-allergic and anti-inflammatory effects, but have a problem of strong side effects. Indomethacin and the like have only anti-inflammatory effects, and thus are not useful as anti-allergic agents.
また、薬物の投与で一時的に症状が抑制されることがあっても、効果および副作用の面で、充分とはいえない実状にある。かかる現状において、各種アレルギー疾患の中でも、特に気管支ぜん息、アレルギー性鼻炎、花粉症、アトピー性皮膚炎などに対して強い抗アレルギー効果を有し、しかも長時間の投与に耐え得る安全性の高い、かつ経口投与可能な新しい治療薬の開発が望まれている。 Moreover, even if the symptom is temporarily suppressed by the administration of a drug, it is in reality that it is not sufficient in terms of effects and side effects. Under such circumstances, among allergic diseases, it has a strong antiallergic effect especially against bronchial asthma, allergic rhinitis, hay fever, atopic dermatitis and the like, and is highly safe to withstand long-term administration. In addition, development of new therapeutic agents that can be administered orally is desired.
一方、ミカン科植物のＣｉｔｒｕｓ属植物やＦｏｒｔｕｎｅｌｌａ属植物の果実は柑橘類果実と総称され、世界的に食用とされているものが数多くある。また、柑橘類果実は古来、薬用に供されるものも多く、『第十四改正日本薬局方』には陳皮［ミカン科
（Rutaceae）のウンシュウミカン（Citrus unshiu）あるいはCitrus reticulataの成熟した果皮］、枳実・枳殻［ミカン科（Rutaceae）のダイダイCitrus aurantium var.
daidai、Citrus aurantium又はナツミカンCitrus natsudaidaiの未熟果をそのまま或いはそれを半分に横切したもの］、橙皮［ミカン科（Rutaceae）のCitrus aurantium又はダイダイCitrus aurantium var. daidaiの成熟した果皮］が収載されている。
On the other hand, the fruits of Citrus plants and Fortunella plants of the Citrus family are collectively called citrus fruits, and there are many that are edible worldwide. Citrus fruits have been used for medicinal purposes since ancient times, and the 14th revised Japanese pharmacopoeia includes cinnamon [Rutaceae (Citrus unshiu) or Citrus reticulata mature pericarp], Fruit berry, rice husk [Rutaceae, Daidai Citrus aurantium var.
Ripe daidai, Citrus aurantium or Natsumikan Citrus natsudaidai immature fruit as it is or cut in half, orange peel [Rutaceae Citrus aurantium or Daidai Citrus aurantium var. ing.
これら柑橘系生薬はいずれも特有の芳香と苦味を有し、中国では、陳皮および橙皮は芳香性苦味健胃薬として用いられ、枳実は堅く充実したうっ滞、うっ血による腫脹の改善を目的とした漢方方剤に配合される。 All of these citrus herbal medicines have a unique aroma and bitterness. In China, Chen and orange peels are used as aromatic bitter stomachic medicines, and the fruit is intended to improve solid and solid stasis and swelling caused by congestion. Formulated in Chinese medicine.
柑橘系生薬には、精油成分としてリモネン（limonene）、フラボノイド配糖体としてヘスペリジン（hesperidin）、ネオヘスペリジン（neohesperidin）、ナリンギン
（naringin）、アルカロイドとしてシネフリン（synephrine）などが含まれている。
Citrus herbal medicines include limonene as an essential oil component, hesperidin, neohesperidin, naringin as a flavonoid glycoside, and synephrine as an alkaloid.
ミカン類生薬（陳皮、橘皮、枳実、橙皮）には、消化器系に対する作用や抗アレルギー作用などが証明されている。抗アレルギー作用に関しては、陳皮水製エキスの経口投与によりラット受身皮膚アナフィラキシー反応は抑制される。ウンシュウミカン果実についてもマスト細胞からのｃｏｍｐｏｕｎｄ ４８／８０によるヒスタミン遊離抑制作用を指標に、抗アレルギー作用を検討され、成熟したものよりも未成熟なものにより強い作用が認められている。たとえば、特許文献１参照。また、ヘスペリジンが主要成分と考えられている。たとえば、特許文献２参照。また、フラボノイド成分のノビレチン、タンゲリチン、３−メトキシノビレチンにもラット腹腔マスト細胞からのヒスタミン遊離抑制作用が認められている。また、枳実、枳殻の水エキスや５０％エタノールエキスにはラット受身皮膚アナフィラキシー反応やマウス塩化ピクリル誘発接触性皮膚炎を抑制する作用が報告されている。 Citrus herbal medicines (Chen bark, tachibana bark, persimmon, orange peel) have been proven to have an action on the digestive system and an antiallergic action. With regard to antiallergic action, rat passive skin anaphylactic reaction is suppressed by oral administration of an extract made from Chensuisui. Anti-allergic action has also been examined on the histamine release inhibitory effect of compound 48/80 from mast cells as an index for Satsuma mandarin fruit, and a stronger action is recognized on immature fruit than on mature fruit. For example, see Patent Document 1. Hesperidin is also considered a major component. For example, see Patent Document 2. In addition, flavonoid components nobiletin, tangelitin, and 3-methoxynobiletin have been found to inhibit histamine release from rat peritoneal mast cells. Moreover, the effect which suppresses rat passive skin anaphylactic reaction and mouse | mouth picryl chloride induced contact dermatitis has been reported to the fruit extract, the rice husk water extract, and 50% ethanol extract.
特許文献１記載の技術は、有効成分として柑橘類の未成熟な果実の水溶性エキスを含有してなる抗アレルギー剤を提供するものである。特許文献２記載の技術はヘスペリジンを有効成分としてなる抗アレルギー剤を提供するものである。しかし、ヘスペリジンは極めて水に難溶性故、柑橘類の未成熟な果実を水で抽出した抗アレルギー剤に抗アレルギー効果は期待できない。したがって、エキス中に多量のヘスペリジンを含有してなる抗アレルギー剤が求められている。 The technique described in Patent Document 1 provides an antiallergic agent comprising a water-soluble extract of citrus immature fruit as an active ingredient. The technique described in Patent Document 2 provides an antiallergic agent containing hesperidin as an active ingredient. However, since hesperidin is extremely insoluble in water, an antiallergic effect cannot be expected from an antiallergic agent obtained by extracting immature citrus fruits with water. Therefore, an antiallergic agent comprising a large amount of hesperidin in the extract is required.
本発明の目的は、抗アレルギー効果に優れ、気管支ぜん息、アレルギー性鼻炎、花粉症、アトピー性皮膚炎などのアレルギー性疾患の予防・改善・治療に有効でかつ安全な抗アレルギー剤を提供することにある。 An object of the present invention is to provide an antiallergic agent that is excellent in antiallergic effect, effective and safe for preventing, ameliorating, and treating allergic diseases such as bronchial asthma, allergic rhinitis, hay fever, and atopic dermatitis. It is in.
かかる事情により、本発明者らは鋭意研究を行った結果、ヘスペリジンを含有する未成熟な柑橘系果実のエタノール抽出エキスあるいは乾燥粉末がジニトロフルオロベンゼン誘発３相性皮膚反応試験において極めて強い抗アレルギー作用を示すことを見出した。 Under these circumstances, as a result of intensive studies, the present inventors have found that an ethanol extract or dried powder of immature citrus fruits containing hesperidin has an extremely strong antiallergic action in a dinitrofluorobenzene-induced three-phase skin reaction test. Found to show.
本発明のヘスペリジンを含有する未成熟な柑橘系果実とは、一般に柑橘系果実の着果後の生長過程において、果皮が黄変する以前の未成熟段階の果実を意味するものである。たとえば、実施例で実験材料に供した和歌山有田郡金屋町産のウンシュウミカンにおいては、５月２０日頃に着果し、その後２ヶ月ないし３ヶ月を経過したウンシュウミカンにおいては、着果後２カ月ないし３カ月を経過した横径が約２．０ｃｍ以上、約４．５ｃｍ以下の未成熟な果実がこれに相当する。 The immature citrus fruit containing hesperidin of the present invention generally means a fruit in an immature stage before the skin peels yellow in the growth process after fruiting of the citrus fruit. For example, in Satsuma mandarin from Kanaya-cho, Arita-gun, Wakayama, which was used as experimental materials in the examples, the fruit arrived around May 20th, and in Satsuma mandarin after two to three months, two months after fruit set. This corresponds to an immature fruit having a lateral diameter of about 2.0 cm or more and about 4.5 cm or less after 3 months.
本発明のエタノール抽出エキスは、エタノールを含有した水溶液で抽出するエキスである。本発明においては、１００％エタノールも４０％以上のエタノール水溶液に含めるものとする。６０％以上のエタノール水溶液の場合も同様である。 The ethanol extract of the present invention is an extract extracted with an aqueous solution containing ethanol. In the present invention, 100% ethanol is also included in 40% or more ethanol aqueous solution. The same applies to a 60% or more ethanol aqueous solution.
本発明のヘスペリジン含有未成熟柑橘系果実のエキスは、一般の抽出方法によって得られる。即ち、果実を細切後、１０倍量の４０％以上のエタノールで抽出、ろ過後、そのろ液を減圧下でエタノールを留去後、抽出エキスを得る。また、ヘスペリジン含有未成熟柑橘系果実の乾燥粉末は、一般の乾燥方法によって得られる。すなわち、果実をそのままあるいは細切後、日陰あるいは陽干し、あるいは乾燥機を用いて乾燥し、粉末としたものである。 The hesperidin-containing immature citrus fruit extract of the present invention can be obtained by a general extraction method. That is, after chopping the fruit, extraction with 10 times the amount of 40% or more ethanol and filtration, the filtrate is evaporated under reduced pressure to obtain an extract. Moreover, the dry powder of a hesperidin containing immature citrus fruit is obtained by a general drying method. That is, the fruit is directly or after being shredded, shaded or dried, or dried using a dryer to obtain a powder.
このようにして、得られるエキスあるいは粉末は、そのままあるいは必要に応じて他の公知の添加剤、たとえば、賦形剤、崩壊剤、結合剤、滑沢剤、抗酸化剤、コーティング剤、着色剤、矯味矯臭剤、界面活性剤、可塑剤などを混合して常法により、顆粒剤、散剤、カプセル剤、錠剤、ドライシロップ剤、液剤などの経口剤とすることができる。 In this way, the extract or powder obtained can be used as it is or if necessary, other known additives such as excipients, disintegrants, binders, lubricants, antioxidants, coating agents, colorants. Oral agents such as granules, powders, capsules, tablets, dry syrups, and liquids can be prepared by mixing conventional flavoring agents, surfactants, plasticizers, and the like.
本発明の抗アレルギー剤の有効成分であるヘスペリジン含有未成熟柑橘系果実のエキスあるいは粉末は、１日０．１ｇ〜２０ｇ用いる。特に１日０．５ｇから５ｇ用いることが好ましい。 The extract or powder of hesperidin-containing immature citrus fruit, which is an active ingredient of the antiallergic agent of the present invention, is used in an amount of 0.1 to 20 g per day. It is particularly preferable to use 0.5 to 5 g per day.
本発明は、ヘスペリジンを含有する未成熟な柑橘類果実をエタノール濃度が４０％以上のエタノール水溶液で抽出することにより得られたエキスを含有することを特徴とする抗アレルギー剤である。 The present invention is an antiallergic agent comprising an extract obtained by extracting immature citrus fruit containing hesperidin with an aqueous ethanol solution having an ethanol concentration of 40% or more.
本発明に従えば、未成熟な柑橘類果実をエタノール濃度が４０％以上のエタノールで抽出する。水の場合と比較してエタノール濃度が４０％以上であると抽出されるエキス中にヘスペリジンが増加する。したがって、抗アレルギー効果に優れた抗アレルギー剤を提供することができる。 According to the present invention, immature citrus fruits are extracted with ethanol having an ethanol concentration of 40% or more. When the ethanol concentration is 40% or more compared to the case of water, hesperidin increases in the extracted extract. Therefore, an antiallergic agent having an excellent antiallergic effect can be provided.
また本発明は、ヘスペリジンを含有する未成熟な柑橘類果実をエタノール濃度が６０％以上のエタノール水溶液で抽出することにより得られたエキスを含有することを特徴とする抗アレルギー剤である。 Moreover, this invention is an antiallergic agent characterized by including the extract obtained by extracting the immature citrus fruit containing hesperidin with ethanol aqueous solution whose ethanol concentration is 60% or more.
本発明に従えば、未成熟な柑橘類果実をエタノール濃度が６０％以上のエタノールで抽出する。エタノール濃度が６０％未満の場合と比較してエタノール濃度が６０％以上であると抽出されるエキス中に急激にヘスペリジン含有量が増加する。したがって、抗アレルギー効果に優れた抗アレルギー剤を提供することができる。 According to the present invention, immature citrus fruits are extracted with ethanol having an ethanol concentration of 60% or more. Compared with the case where the ethanol concentration is less than 60%, the hesperidin content increases rapidly in the extracted extract when the ethanol concentration is 60% or more. Therefore, an antiallergic agent having an excellent antiallergic effect can be provided.
また本発明は、ヘスペリジンを含有する未成熟な柑橘類果実の乾燥粉末を含有することを特徴とする抗アレルギー剤である。 Moreover, this invention is an antiallergic agent characterized by including the dry powder of the immature citrus fruit containing hesperidin.
本発明に従えば、ヘスペリジンを含有する未成熟な柑橘類果実の乾燥粉末を有効成分として含有するので、抗アレルギー効果に優れた抗アレルギー剤を提供することができる。 According to the present invention, since an immature citrus fruit dry powder containing hesperidin is contained as an active ingredient, an antiallergic agent having an excellent antiallergic effect can be provided.
また本発明は、未成熟な柑橘類果実をエタノール濃度が４０％以上のエタノール水溶液で抽出する工程を含むことを特徴とする抗アレルギー剤エキスの抽出方法である。 The present invention is also a method for extracting an anti-allergic agent extract, comprising a step of extracting immature citrus fruits with an ethanol aqueous solution having an ethanol concentration of 40% or more.
本発明に従えば、未成熟な柑橘類果実をエタノール濃度が４０％以上のエタノール水溶液でエキスを抽出する。水で抽出する場合と比較してエタノール濃度が４０％以上であると抽出されるエキス中にヘスペリジン含有量が増加する。 According to the present invention, an immature citrus fruit is extracted with an aqueous ethanol solution having an ethanol concentration of 40% or more. Compared with the case of extraction with water, the hesperidin content increases in the extracted extract when the ethanol concentration is 40% or more.
また本発明は、未成熟な柑橘類果実をエタノール濃度が６０％以上のエタノール水溶液で抽出する工程を含むことを特徴とする抗アレルギー剤エキスの抽出方法である。 The present invention is also a method for extracting an anti-allergic agent extract, comprising a step of extracting immature citrus fruits with an aqueous ethanol solution having an ethanol concentration of 60% or more.
本発明に従えば、未成熟な柑橘類果実をエタノール濃度が６０％以上のエタノール水溶液でエキスを抽出する。エタノール濃度が６０％未満の場合と比較してエタノール濃度が６０％以上であると抽出されるエキス中に急激にヘスペリジン含有量が増加する。 According to the present invention, an extract is extracted from an immature citrus fruit with an aqueous ethanol solution having an ethanol concentration of 60% or more. Compared with the case where the ethanol concentration is less than 60%, the hesperidin content increases rapidly in the extracted extract when the ethanol concentration is 60% or more.
また本発明は、柑橘類果実がウンシュウミカンであることを特徴とする。
本発明に従えば、未成熟なウンシュウミカンにはヘスペリジンが多く含まれている。したがって、抗アレルギー効果に優れたヘスペリジンを多量に含有する抗アレルギー剤を提供することができる。
Moreover, the present invention is characterized in that the citrus fruit is Satsuma mandarin.
According to the present invention, immature Satsuma mandarin is rich in hesperidin. Therefore, an antiallergic agent containing a large amount of hesperidin having an excellent antiallergic effect can be provided.
また本発明は、柑橘類果実がウンシュウミカンであることを特徴とする。
本発明に従えば、未成熟なウンシュウミカンにはヘスペリジンが多く含まれている。したがって、抗アレルギー効果に優れたヘスペリジンを多量に含有するエキスを抽出することができる。
Moreover, the present invention is characterized in that the citrus fruit is Satsuma mandarin.
According to the present invention, immature Satsuma mandarin is rich in hesperidin. Therefore, an extract containing a large amount of hesperidin having an excellent antiallergic effect can be extracted.
また本発明は、未成熟な柑橘類果実が、５月下旬に着果した後２カ月ないし３カ月を経過した横径が２．０ｃｍ以上、４．５ｃｍ以下の未成熟なウンシュウミカンであることを特徴とする。 Further, the present invention is that immature citrus fruits are immature Satsuma mandarin having a lateral diameter of 2.0 cm or more and 4.5 cm or less after 2 to 3 months have passed since the end of May. Features.
本発明に従えば、５月下旬に着果した時点から２カ月ないし３カ月を経過した横径が２．０ｃｍ以上、４．５ｃｍ以下の未成熟なウンシュウミカンには多量のヘスペリジンが含まれている。したがって、抗アレルギー効果に優れたヘスペリジンを多量に含有する抗アレルギー剤を提供することができる。 According to the present invention, immature Satsuma mandarin with a horizontal diameter of 2.0 cm or more and 4.5 cm or less after 2 to 3 months of fruiting in late May contains a large amount of hesperidin. Yes. Therefore, an antiallergic agent containing a large amount of hesperidin having an excellent antiallergic effect can be provided.
また本発明は、未成熟な柑橘類果実が、５月下旬に着果した後２カ月ないし３カ月を経過した横径が２．０ｃｍ以上、４．５ｃｍ以下の未成熟なウンシュウミカンであることを特徴とする。 Further, the present invention is that immature citrus fruits are immature Satsuma mandarin having a lateral diameter of 2.0 cm or more and 4.5 cm or less after 2 to 3 months have passed since the end of May. Features.
本発明に従えば、５月下旬に着果した時点から２カ月ないし３カ月を経過した横径が２．０ｃｍ以上、４．５ｃｍ以下の未成熟なウンシュウミカンには多量のヘスペリジンが含まれている。したがって、抗アレルギー効果に優れたヘスペリジンを多量に含有するエキスを抽出することができる。 According to the present invention, immature Satsuma mandarin with a horizontal diameter of 2.0 cm or more and 4.5 cm or less after 2 to 3 months of fruiting in late May contains a large amount of hesperidin. Yes. Therefore, an extract containing a large amount of hesperidin having an excellent antiallergic effect can be extracted.
本発明によれば、本発明の抗アレルギー剤の有効成分であるヘスペリジン含有未成熟柑橘系果実の４０％のエタノール抽出エキスは、抗アレルギー作用を有し、３相性皮膚反応試験に惹起されるアレルギー性の炎症像を抑制するので、アトピー性皮膚炎、花粉症、気管支ぜん息などのアレルギー性疾患の改善、治療に有効である。 According to the present invention, 40% ethanol extract of hesperidin-containing immature citrus fruit, which is an active ingredient of the antiallergic agent of the present invention, has an antiallergic action and is an allergy caused by a three-phase skin reaction test. It is effective in improving and treating allergic diseases such as atopic dermatitis, hay fever and bronchial asthma.
本発明によれば、本発明の抗アレルギー剤の有効成分であるヘスペリジン含有未成熟柑橘系果実の６０％のエタノール抽出エキスは、抗アレルギー作用を有し、３相性皮膚反応試験に惹起されるアレルギー性の炎症像を抑制するので、アトピー性皮膚炎、花粉症、気管支ぜん息などのアレルギー性疾患の改善、治療に有効である。 According to the present invention, 60% ethanol extract of hesperidin-containing immature citrus fruit, which is an active ingredient of the antiallergic agent of the present invention, has an antiallergic action and is an allergy caused by a three-phase skin reaction test. It is effective in improving and treating allergic diseases such as atopic dermatitis, hay fever and bronchial asthma.
本発明によれば、本発明の抗アレルギー剤の有効成分であるヘスペリジン含有未成熟柑橘系果実の粉末は、抗アレルギー作用を有し、３相性皮膚反応試験に惹起されるアレルギー性の炎症像を抑制するので、アトピー性皮膚炎、花粉症、気管支ぜん息などのアレルギー性疾患の改善、治療に有効である。 According to the present invention, the hesperidin-containing immature citrus fruit powder, which is the active ingredient of the antiallergic agent of the present invention, has an antiallergic action and exhibits allergic inflammation caused by a three-phase skin reaction test. It is effective for the improvement and treatment of allergic diseases such as atopic dermatitis, hay fever and bronchial asthma.
本発明によれば、抗アレルギー作用を有し、３相性皮膚反応試験に惹起されるアレルギー性の炎症像を抑制する成分の１つであるヘスペリジンを含んだエキスを抽出することができる。 According to the present invention, it is possible to extract an extract containing hesperidin, which is one of the components that have an antiallergic action and suppress allergic inflammation caused by a three-phase skin reaction test.
本発明によれば、抗アレルギー作用を有し、３相性皮膚反応試験に惹起されるアレルギー性の炎症像を抑制する成分の１つであるヘスペリジンを多量に含んだエキスを抽出することができる。 ADVANTAGE OF THE INVENTION According to this invention, the extract which has an antiallergic action and contains hesperidin which is one of the components which suppress the allergic inflammation image induced by the three-phase skin reaction test can be extracted.
本発明によれば、未成熟なウンシュウミカンにはヘスペリジンが多く含まれている。したがって、抗アレルギー効果に優れたヘスペリジンを多量に含有する抗アレルギー剤を提供することができる。 According to the present invention, immature Satsuma mandarin is rich in hesperidin. Therefore, an antiallergic agent containing a large amount of hesperidin having an excellent antiallergic effect can be provided.
本発明によれば、未成熟なウンシュウミカンにはヘスペリジンが多く含まれている。したがって、抗アレルギー効果に優れたヘスペリジンを多量に含有するエキスを抽出することができる。 According to the present invention, immature Satsuma mandarin is rich in hesperidin. Therefore, an extract containing a large amount of hesperidin having an excellent antiallergic effect can be extracted.
本発明によれば、５月下旬に着果した時点から２カ月ないし３カ月を経過した横径が２．０ｃｍ以上、４．５ｃｍ以下の未成熟なウンシュウミカンには多量のヘスペリジンが含まれている。したがって、抗アレルギー効果に優れたヘスペリジンを多量に含有する抗アレルギー剤を提供することができる。 According to the present invention, immature Satsuma mandarin with a horizontal diameter of 2.0 cm or more and 4.5 cm or less after 2 to 3 months from the end of May contains a large amount of hesperidin. Yes. Therefore, an antiallergic agent containing a large amount of hesperidin having an excellent antiallergic effect can be provided.
本発明によれば、５月下旬に着果した時点から２カ月ないし３カ月を経過した横径が２．０ｃｍ以上、４．５ｃｍ以下の未成熟なウンシュウミカンには多量のヘスペリジンが含まれている。したがって、抗アレルギー効果に優れたヘスペリジンを多量に含有するエキスを抽出することができる。 According to the present invention, immature Satsuma mandarin with a horizontal diameter of 2.0 cm or more and 4.5 cm or less after 2 to 3 months from the end of May contains a large amount of hesperidin. Yes. Therefore, an extract containing a large amount of hesperidin having an excellent antiallergic effect can be extracted.
次に、実施例を挙げて本発明をさらに説明するが、本発明はこれら実施例に限定されるものではない。 EXAMPLES Next, although an Example is given and this invention is further demonstrated, this invention is not limited to these Examples.
試験例１
ジニトロフルオロベンゼン（ＤＮＦＢ）誘発３相性皮膚反応に及ぼす未成熟なウンシュウミカン果実の影響
Test example 1
Effects of immature Satsuma mandarin fruit on dinitrofluorobenzene (DNFB) -induced triphasic skin reaction
検体１；２００３年、８月５日、和歌山県有田郡金屋町で栽培されている未成熟なウンシュウミカン果実を採取、細切後乾燥し、さらに粉砕、粉末とし、被検体に供した。 Specimen 1; On August 5, 2003, immature citrus fruit cultivated in Kanaya-cho, Arita-gun, Wakayama Prefecture was collected, shredded, dried, ground and powdered, and used as a specimen.
検体２；２００３年、８月５日、和歌山県有田郡金屋町で栽培されている未成熟なウンシュウミカン果実を採取、細切後乾燥し、粉砕後、１０倍量の水で２時間、２回熱時（約１００℃）抽出した。抽出液を熱時ろ過し、ろ液を減圧下に濃縮し、凍結乾燥を施して得た乾燥水抽出エキス（収率；３４．０％）を被検体に供した。 Specimen 2; Immature Satsuma mandarin fruit cultivated in Kanaya-cho, Arita-gun, Wakayama, August 5th, 2003, collected after being shredded, dried, crushed, and 2 hours for 2 hours with 10 times the amount of water Extraction was performed during reheating (about 100 ° C.). The extract was filtered while hot, the filtrate was concentrated under reduced pressure, and a dry water extract (yield; 34.0%) obtained by lyophilization was used as a specimen.
検体３；検体２の水を２０％エタノールで熱時（約８５℃）抽出した以外は同様にして得られたエキス（収率；３６．０％）。 Sample 3: Extract obtained in the same manner except that the water of Sample 2 was extracted with 20% ethanol when heated (about 85 ° C.) (yield: 36.0%).
検体４；検体２の水を４０％エタノールで熱時（約８５℃）抽出した以外は同様にして得られたエキス（収率；３８．０％）。 Sample 4: Extract obtained in the same manner except that the water of Sample 2 was extracted with 40% ethanol when heated (about 85 ° C.) (yield: 38.0%).
検体５；検体２の水を６０％エタノールで熱時（約８０℃）抽出した以外は同様にして得られたエキス（収率；３９．７％）。 Sample 5: Extract obtained in the same manner except that the water of Sample 2 was extracted with 60% ethanol when heated (about 80 ° C.) (yield: 39.7%).
検体６；検体２の水をエタノールで熱時（約８０℃）抽出した以外は同様にして得られたエキス（収率；３４．９％）。 Sample 6: Extract obtained in the same manner except that the water of Sample 2 was extracted with ethanol (about 80 ° C.) when heated (yield: 34.9%).
（試験方法）
１）ジニトロフェニル化卵白アルブミン（ＤＮＰ−ＯＶＡ）の調製；卵白アルブミン（ovalbumin、type V）およびＫ２ＣＯ３を各２ｇずつ１００ｍｌの水に溶解し、この溶液に２ｇのジニトロベンゼンスルホン酸ナトリウム塩（dinitrobenzenesulfonic acid
sodium salt）を加え、遮光下で３７℃にて、２４時間スターラーで攪拌し、得られた反応液を水で２日間透析後、その内液を凍結乾燥し、調製した。
(Test method)
1) Preparation of dinitrophenylated ovalbumin (DNP-OVA); 2 g of ovalbumin (type V) and K 2 CO 3 were dissolved in 100 ml of water, and 2 g of dinitrobenzenesulfonic acid sodium salt was dissolved in this solution. (Dinitrobenzenesulfonic acid
sodium salt) was added, and the mixture was stirred with a stirrer at 37 ° C. for 24 hours under light shielding. The resulting reaction solution was dialyzed with water for 2 days, and the internal solution was freeze-dried to prepare.
２）ＤＮＦＢ誘発３相性皮膚反応試験；ＩＣＲ系雌性マウスにゲル状の水酸化アルミニウム（aluminumhydroxide gel） １ｍｇとＤＮＰ−ＯＶＡ １０μｇを含む生理食塩液０．２ｍＬを腹腔内投与し、能動的に感作する。その１週間後、０．１％ ＤＮＦＢエタノール溶液を両耳の表裏に１０μｌずつ塗布し、誘発２４時間後に充分に腫脹しているマウス（浮腫率１０％以上）を選別する。その後、選別したマウスに再度感作し、１週間後に反応を惹起する。耳介の厚さは反応惹起前および惹起後１、２４時間および８日後に
dial thickness gaugeを用いて測定し、耳介浮腫率として算定する。反応を惹起した時点から１時間後の耳介浮腫率をＩＰＲ（Immediate Phase Response）浮腫率、２４時間後の耳介浮腫率をＬＰＲ(Late Phase Response)浮腫率、８日後の耳介浮腫率をｖＬＰＲ(very
Late Phase Response)浮腫率とする。なお、被検体は反応惹起１時間前および惹起２日後から８日後まで連日経口投与する。被検体および陽性対照薬は、０．２％ＣＭＣ・Ｎａに溶解あるいは懸濁し、ウンシュウミカン粉末は水に懸濁し、マウス体重１０ｇあたり０．２ｍＬの用量で経口投与した。
2) DNFB-induced triphasic skin reaction test; ICR female mice were intraperitoneally administered with 0.2 mL of physiological saline containing 1 mg of aluminum hydroxide oxide gel and 10 μg of DNP-OVA, and actively sensitized To do. One week later, 10 μl of 0.1% DNFB ethanol solution is applied to the front and back of both ears, and mice that are sufficiently swollen 24 hours after induction (edema rate of 10% or more) are selected. Thereafter, the selected mouse is sensitized again, and a reaction is induced after one week. The thickness of the auricles was measured before the reaction and after 1, 24 hours and 8 days after the reaction.
Measured using a dial thickness gauge and calculated as a pinna edema rate. The auricular edema rate 1 hour after the reaction was triggered was the IPR (Immediate Phase Response) edema rate, the auricular edema rate 24 hours later was the LPR (Late Phase Response) edema rate, and the auricular edema rate 8 days later. vLPR (very
Late Phase Response) The edema rate. The subject is orally administered every day from 1 hour before reaction initiation and from 2 days to 8 days after induction. The subject and the positive control drug were dissolved or suspended in 0.2% CMC · Na, and the Citrus unshiu powder was suspended in water and orally administered at a dose of 0.2 mL per 10 g of mouse body weight.
（結果）結果を表１に示す。
表１は、３相性皮膚反応に及ぼす未成熟ウンシュウミカン果実の影響を示す。
(Results) The results are shown in Table 1.
Table 1 shows the effect of immature Satsuma mandarin fruit on the triphasic skin reaction.
マウスに３相性皮膚反応を惹起させたところ、ＩＰＲにおいては１９．７±１．３％、ＬＰＲにおいては１８．１±１．２％、ｖＬＰＲにおいては１９．７±０．９％の耳浮腫が観察された。検体１は２００ｍｇ／ｋｇ用量の経口投与で、いずれの耳浮腫も有意に抑制した。未成熟なウンシュウミカン果実を水で抽出した検体２はいずれの耳浮腫に対しても有意な抑制作用が認められなかった。ウンシュウミカン果実を２０％のエタノールで抽出した検体２においては、ＩＰＲとＬＰＲの耳浮腫を有意に抑制したが、ｖＬＰＲには無効であった。ウンシュウミカン果実を４０％以上のエタノールで抽出すると、いずれの浮腫に対しても有意な抑制作用がみられる。その作用強度はエタノールの含量が高まるほど強まった。特に６０％のエタノールで抽出した検体５では検体４に比べて急激に作用強度が強まった。 When a triphasic skin reaction was induced in mice, ear edema of 19.7 ± 1.3% in IPR, 18.1 ± 1.2% in LPR, and 19.7 ± 0.9% in vLPR Was observed. Specimen 1 was orally administered at a dose of 200 mg / kg and significantly suppressed any ear edema. Specimen 2 obtained by extracting immature Satsuma mandarin fruit with water showed no significant inhibitory action against any ear edema. In Sample 2 obtained by extracting Satsuma mandarin fruit with 20% ethanol, ear edema of IPR and LPR was significantly suppressed, but it was ineffective for vLPR. Extraction of Citrus unshiu fruits with 40% or more of ethanol shows a significant inhibitory effect on any edema. The strength of action increased as the ethanol content increased. In particular, the sample 5 extracted with 60% ethanol showed a stronger intensity of action than the sample 4.
試験例２
ジニトロフルオロベンゼン（ＤＮＦＢ）誘発３相性皮膚反応に及ぼすヘスペリジンの影響
Test example 2
Effect of hesperidin on triphasic skin reaction induced by dinitrofluorobenzene (DNFB)
検体１；２００３年、８月５日、和歌山県有田郡金屋町で栽培されている未成熟なウンシュウミカン果実を採取、その後細切、乾燥した。乾燥未成熟ウンシュウミカン果実から、松田ら［薬学雑誌、１１１、１９３−１９８（１９９１）］の方法でヘスペリジンを単離・同定し、被検体に供した。 Specimen 1; On August 5, 2003, immature Satsuma mandarin fruit cultivated in Kanaya-cho, Arita-gun, Wakayama Prefecture was collected, then chopped and dried. Hesperidin was isolated and identified from dried immature Satsuma mandarin fruit by the method of Matsuda et al. [Pharmaceutical Journal, 111, 193-198 (1991)], and was used as a subject.
（試験方法）
１）ジニトロフェニル化卵白アルブミン（ＤＮＰ−ＯＶＡ）の調製；卵白アルブミン（ovalbumin、type V）およびＫ２ＣＯ３を各２ｇずつ１００ｍｌの水に溶解し、この溶液に２ｇのジニトロベンゼンスルホン酸ナトリウム塩（dinitrobenzenesulfonic acid
sodium salt）を加え、遮光下で３７℃にて、２４時間スターラーで攪拌し、得られた反応液を水で２日間透析後、その内液を凍結乾燥し、調製した。
(Test method)
1) Preparation of dinitrophenylated ovalbumin (DNP-OVA); 2 g of ovalbumin (type V) and K 2 CO 3 were dissolved in 100 ml of water, and 2 g of dinitrobenzenesulfonic acid sodium salt was dissolved in this solution. (Dinitrobenzenesulfonic acid
sodium salt) was added, and the mixture was stirred with a stirrer at 37 ° C. for 24 hours under light shielding. The resulting reaction solution was dialyzed with water for 2 days, and the internal solution was freeze-dried to prepare.
２）ＤＮＦＢ誘発３相性皮膚反応試験；ＩＣＲ系雌性マウスにゲル状の水酸化アルミニウム（aluminumhydroxide gel） １ｍｇとＤＮＰ−ＯＶＡ １０μｇを含む生理食塩液０．２ｍＬを腹腔内投与し、能動的に感作する。その１週間後、０．１％ＤＮＦＢエタノール溶液を両耳の表裏に１０μｌずつ塗布し、誘発２４時間後に充分に腫脹しているマウス（浮腫率１０％以上）を選別する。その後、選別したマウスに再度感作し、１週間後に反応を惹起する。耳介の厚さは反応惹起前および惹起後１、２４時間および８日後にdial thickness gaugeを用いて測定し、耳介浮腫率として算定する。なお、被検体は反応惹起１時間前および惹起２日後から８日後まで連日経口投与する。被検体および陽性対照薬は、０．２％ＣＭＣ・Ｎａに溶解あるいは懸濁し、ウンシュウミカン粉末は水に懸濁し、マウス体重１０ｇあたり０．２ｍＬの用量で経口投与した。 2) DNFB-induced triphasic skin reaction test; ICR female mice were intraperitoneally administered with 0.2 mL of physiological saline containing 1 mg of aluminum hydroxide oxide gel and 10 μg of DNP-OVA, and actively sensitized To do. One week later, 10 μl of 0.1% DNFB ethanol solution is applied to both front and back of both ears, and mice that are sufficiently swollen 24 hours after induction (edema rate of 10% or more) are selected. Thereafter, the selected mouse is sensitized again, and a reaction is induced after one week. The thickness of the auricle is measured using a dial thickness gauge before the reaction is induced and after 1, 24 hours and 8 days after the reaction is induced, and is calculated as a pinna edema rate. The subject is orally administered every day from 1 hour before reaction initiation and from 2 days to 8 days after induction. The subject and the positive control drug were dissolved or suspended in 0.2% CMC · Na, and the Citrus unshiu powder was suspended in water and orally administered at a dose of 0.2 mL per 10 g of mouse body weight.
（結果）結果を表２に示す。
表２は、３相性皮膚反応に及ぼすヘスペリジンの影響を示す。
(Results) The results are shown in Table 2.
Table 2 shows the effect of hesperidin on the triphasic skin reaction.
ヘスペリジンは２０ｍｇ／ｋｇおよび１００ｍｇ／ｋｇの用量で、ＤＮＦＢによって誘発される３相性皮膚反応による耳浮腫を有意に抑制する作用を示した。１００ｍｇ／ｋｇ用量のヘスペリジンの作用強度はステロイド剤のプレゾニゾロン １０ｍｇ／ｋｇ用量の抑制作用とほぼ同程度であった。 Hesperidin showed a significant inhibitory effect on ear edema due to a triphasic skin reaction induced by DNFB at doses of 20 mg / kg and 100 mg / kg. The potency of hesperidin at the 100 mg / kg dose was almost the same as the inhibitory effect of the steroidal prezonizolone at 10 mg / kg.
試験例３
未成熟なウンシュウミカン果実エキスおよび乾燥果実中のヘスペリジン含量の測定
Test example 3
Determination of hesperidin content in immature Satsuma mandarin fruit extract and dried fruit
検体１；２００３年、８月５日、和歌山県有田郡金屋町で栽培されている未成熟なウンシュウミカン果実を採取乾燥し、粉砕後、粉末とした。この粉末１０ｇに１００ｍＬのメタノールを加え、２時間、２回熱時抽出した。抽出液を熱時ろ過し、ろ液を減圧下に濃縮し、凍結乾燥を施して得た乾燥メタノール抽出エキス（収率；４１．０％）を被検体とした。 Specimen 1: immature Unshu mandarin fruit cultivated in Kanaya-cho, Arita-gun, Wakayama, on August 5, 2003, was collected, dried, ground, and powdered. 100 mL of methanol was added to 10 g of this powder, and the mixture was extracted with heating twice for 2 hours. The extract was filtered while hot, the filtrate was concentrated under reduced pressure, and freeze-dried dry methanol extract (yield: 41.0%) was used as the specimen.
検体２；２００３年、８月５日、和歌山県有田郡金屋町で栽培されている未成熟なウンシュウミカン果実を採取、細切後乾燥し、粉砕後、１０倍量の水で２時間、２回熱時抽出した。抽出液を熱時ろ過し、ろ液を減圧下に濃縮し、凍結乾燥を施して得た乾燥水抽出エキス（収率；３４．０％）を被検体に供した。 Specimen 2; Immature Satsuma mandarin fruit cultivated in Kanaya-cho, Arita-gun, Wakayama, August 5th, 2003, collected after being shredded, dried, crushed, and 2 hours for 2 hours with 10 times the amount of water Extracted during recuperation. The extract was filtered while hot, the filtrate was concentrated under reduced pressure, and a dry water extract (yield; 34.0%) obtained by lyophilization was used as a specimen.
検体３；検体２の水を２０％エタノールで抽出した以外は同様にして得られたエキス（収率；３６．０％）。 Sample 3: Extract obtained in the same manner except that the water of Sample 2 was extracted with 20% ethanol (yield: 36.0%).
検体４；検体２の水を４０％エタノールで抽出した以外は同様にして得られたエキス（収率；３８．０％）。 Sample 4: Extract obtained in the same manner except that the water of Sample 2 was extracted with 40% ethanol (yield: 38.0%).
検体５；検体２の水を６０％エタノールで抽出した以外は同様にして得られたエキス（収率；３９．７％）。 Sample 5: Extract obtained in the same manner except that the water of Sample 2 was extracted with 60% ethanol (yield: 39.7%).
検体６；検体２の水をエタノールで抽出した以外は同様にして得られたエキス（収率；３４．９％）。 Sample 6: Extract obtained in the same manner except that the water of Sample 2 was extracted with ethanol (yield: 34.9%).
（試験方法）
各検体を５００ｍｇ精秤した。１００ｍＬのメタノールで３０分、３回還流抽出した（水浴８０℃）。抽出液をろ紙ろ過し、そのろ液をエバポレーターで濃縮した。濃縮した抽出液をメタノールで５０ｍＬにメスアップ後、メタノールで１０倍に希釈した。希釈液をウルトラフリーＭＣ（０．２２ｍｍ）（ＭＩＬＬＩＰＯＲＥ）にてろ過した後、下記の方法でＨＰＬＣ分析を行った。
(Test method)
Each specimen was accurately weighed 500 mg. The mixture was refluxed and extracted three times for 30 minutes with 100 mL of methanol (water bath 80 ° C.). The extract was filtered with filter paper, and the filtrate was concentrated with an evaporator. The concentrated extract was diluted to 50 mL with methanol and diluted 10-fold with methanol. The diluted solution was filtered through Ultra Free MC (0.22 mm) (MILLIPORE) and then subjected to HPLC analysis by the following method.
［ＨＰＬＣ分析条件］
１）検量線の作成；ヘスペリジンの標準品はＳＩＧＭＡ社より購入し、再結晶したものを使用した。メタノールにて１ｍｇ／ｍＬに調製後、２５、５０、１００、２５０、５００（ｍｇ／ｍＬ）に調製し、ウルトラフリーＭＣ（０．２２ｍｍ）（ＭＩＬＬＩＰＯＲＥ）にてろ過したものを検量線作成用の溶液とした。これについて、下記の条件により、ＨＰＬＣ分析を行い、得られたヘスペリジンのピーク面積値から検量線を作成した。
[HPLC analysis conditions]
1) Preparation of calibration curve: Standard hesperidin was purchased from SIGMA and recrystallized. After preparing to 1 mg / mL with methanol, adjust to 25, 50, 100, 250, 500 (mg / mL) and filter with Ultra Free MC (0.22 mm) (MILLIPORE). It was set as the solution. About this, HPLC analysis was performed on the following conditions, and the analytical curve was created from the peak area value of the obtained hesperidin.
２）ＨＰＬＣの分析条件
装置 ；バイナリポンプ：Ｇ１３１２Ａ（Ａｇｉｌｅｎｔ製）
デガッサー；Ｇ１３７７９Ａ ＤＥＧＡＳＳＥＲ（Ａｇｉｌｅｎｔ製）
検出器；ＵＶ−Ｖｉｓ検出器Ｇ１３１４Ａ ＶＷＤ（Ａｇｉｌｅｎｔ製）
カラム恒温槽；Ｇ１３１６Ａ ＣＯＬＣＯＭ（Ａｇｉｌｅｎｔ製）
オートサンプラー；Ｇ１３１３Ａ ＡＬＳ（Ａｇｉｌｅｎｔ製）
カラム；ＹＭＣ−Ｐａｃｋ Ｐｒｏ Ｃ１８（２５０×４．６ｍｍ ｉ．ｄ．）（ＹＭＣ製）
ガードカラム；Ｃ−ＫＧＣ−３２４ＡＳＣ−３（ＹＭＣ製）
移動相；ａｃｅｔｏｎｉｔｒｉｌｅ／１０ｍＭ Ｈ３ＰＯ４（２０：８０（ｖ／ｖ））
流速；１．０ｍＬ／ｍｉｎ
温度；３７℃
検出波長；２８０ｎｍ
注入量；５ｍＬ
データ処理ソフト；Ｘｃａｌｉｂｕｒ（Ｔｈｅｒｍｏ Ｆｉｎｎｉｇａｎ製）ヘスペリジンのＲｅｔｅｎｔｉｏｎ Ｔｉｍｅ；１８．５−１９．５ｍｉｎ
2) Analytical conditions for HPLC Apparatus; Binary pump: G1312A (manufactured by Agilent)
Degasser; G13779A DEGASSER (manufactured by Agilent)
Detector: UV-Vis detector G1314A VWD (manufactured by Agilent)
Column thermostat; G1316A COLCOM (manufactured by Agilent)
Autosampler; G1313A ALS (manufactured by Agilent)
Column; YMC-Pack Pro C18 (250 × 4.6 mm id) (manufactured by YMC)
Guard column; C-KGC-324ASC-3 (manufactured by YMC)
Mobile phase; acetonitrile, 10 mM H 3 PO 4 (20:80 (v / v))
Flow rate: 1.0 mL / min
Temperature: 37 ° C
Detection wavelength: 280 nm
Injection volume: 5 mL
Data processing software: Xcalibur (manufactured by Thermo Finnigan) Hesperidin Retention Time; 18.5 to 19.5 min
（結果）結果を表３に示す。
表３は、未成熟なウンシュウミカン果実からのエキス収率とヘスペリジン含量を示す。
(Results) The results are shown in Table 3.
Table 3 shows the extract yield and hesperidin content from immature Satsuma mandarin fruit.
未成熟なウンシュウミカン果実の乾燥粉末（検体1）をメタノールで抽出し、その含量を測定した結果、１２．１％であった。未成熟なウンシュウミカン果実を水で抽出したエキス（検体２）１ｇ中には１６．４ｍｇのヘスペリジンが含まれ、エキス中の含量から求められる乾燥果実１ｇ中のヘスペリジン含量（％）はわずか０．５６％となり、乾燥果実に含まれるヘスペリジンの約１／２０量しか抽出されていない。抽出溶媒を水からエタノールに変換すると、その濃度は増加するにつれて抽出されてくるヘスペリジン含量が増加する。エタノール濃度が４０％以上であると、水に比べてヘスペリジンの量が明らかに増加している。さらに、エタノール濃度４０％と比較してエタノール濃度６０％では急激にヘスペリジン含量が増加している。１００％エタノールで抽出すると、果実に含まれているヘスペリジンの約１／６量が抽出された。 The dried powder (specimen 1) of immature Satsuma mandarin fruit was extracted with methanol, and the content was measured and found to be 12.1%. 1 g of hesperidin contained in 1 g of an extract of an immature Satsuma mandarin fruit (sample 2) containing 16.4 mg of hesperidin, and the hesperidin content (%) in 1 g of dried fruit obtained from the content in the extract is only 0. 56%, and only about 1/20 of hesperidin contained in the dried fruit is extracted. When the extraction solvent is converted from water to ethanol, the extracted hesperidin content increases as its concentration increases. When the ethanol concentration is 40% or more, the amount of hesperidin is clearly increased as compared with water. Furthermore, the hesperidin content increases rapidly at an ethanol concentration of 60% compared to an ethanol concentration of 40%. When extracted with 100% ethanol, about 1/6 of hesperidin contained in the fruit was extracted.
Claims (9)
The immature Citrus fruit is immature Satsuma mandarin having a lateral diameter of 2.0 cm or more and 4.5 cm or less after two to three months after ripening in late May. 4. Extraction method of antiallergic agent extract of 4 or 5.
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| JP2009544604A (en) * | 2006-07-20 | 2009-12-17 | オーラルデント・リミテッド | Oral compositions and their manufacture and use |
| JP2008137985A (en) * | 2006-11-10 | 2008-06-19 | Univ Kinki | Medicine reinforcing anti-allergic effect without increasing adverse reaction of adrenocortical hormone and method for enhancing anti-allergic effect without increasing adverse reaction of adrenocortical hormone |
| JP2009007256A (en) * | 2007-06-26 | 2009-01-15 | Kao Corp | Nadh/nadph oxidase inhibitor |
| WO2015002393A1 (en) * | 2013-07-03 | 2015-01-08 | 바이오스펙트럼(주) | Composition for treating or preventing inflammatory skin disease, comprising, as active ingredient, immature citrus fruit extract, or synephrine or salt thereof |
| KR101567633B1 (en) * | 2013-07-03 | 2015-11-10 | 바이오스펙트럼 주식회사 | Composition for treatment or prevention of inflammatory skin diseases comprising unripe Citurs unshiu extract, or synephrine or salts thereof |
| CN105358170A (en) * | 2013-07-03 | 2016-02-24 | 韩国百鸥思特公司 | Composition for treating or preventing inflammatory skin disease, comprising, as active ingredient, immature citrus fruit extract, or synephrine or salt thereof |
| US10130670B2 (en) | 2013-07-03 | 2018-11-20 | Biospectrum, Inc. | Composition for treating or preventing inflammatory skin disease, comprising, as active ingredient, immature citrus fruit extract, or synephrine or salt thereof |
| JP2019151568A (en) * | 2018-03-01 | 2019-09-12 | アサヒ飲料株式会社 | Enhancers of acid sphingomyelinase activity, compositions for enhancing acid sphingomyelinase activity and anti-allergic compositions |
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