JP2004292326A - Antiallergic agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an antiallergic agent derived from a natural product or its derivative effectively carrying out prophylaxis or treatment of allergic diseases. <P>SOLUTION: The antiallergic agent comprises a compound represented by formula (1) [wherein, R<SP>1</SP>and R<SP>2</SP>are each independently a hydrogen atom, a methyl group, an ethyl group or an isopropyl group, and R<SP>1</SP>and R<SP>2</SP>together may be a CH<SB>2</SB>; R<SP>3</SP>is a hydroxy group, a CH=CH-(CH<SB>2</SB>)<SB>n</SB>-COOH or an O-sugar; and n is an integer of 0-5] or a pharmaceutically acceptable salt thereof. Furthermore, a medicinal composition, a food additive, a food, a cosmetic, etc., contain the antiallergic agent. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

【００１０】
（式中、Ｒ^１およびＲ^２は、それぞれ独立に、水素原子、メチル基、エチル基、またはイソプロピル基であり、また、Ｒ^１およびＲ^２が一緒になって、−ＣＨ_２−であってもよく、
Ｒ^３は、水酸基、−ＣＨ＝ＣＨ−（ＣＨ_２）_ｎ−ＣＯＯＨ、または−Ｏ−ｓｕｇａｒであり、
ｎは、０から５の整数である）
で表される化合物、または薬学的に許容されうるその塩を含む、抗アレルギー剤が提供される。
ここで、ｓｕｇａｒとは、生体内に存在する糖一般のことであり、単糖、または２〜１０のオリゴ糖であってもよく、例えば、グルコース、ラムノース、ガラクトースのような六単糖やフラクトースのような五単糖で構成される糖類が挙げられる。式（１）で表される化合物としては、例えば、セサモール、カフェー酸、またはそのプロドラックとしての誘導体が挙げられる。薬学的に許容されうるその塩とは、特に限定されるものではないが、例えば、ナトリウム塩、カリウム塩、マグネシウム塩、カルシウム塩などの金属塩、アンモニウム塩、メチルアンモニウム塩またはエチルアンモニウム塩などのモノアルキルアンモニウム塩、ジメチルアンモニウム塩またはジエチルアンモニウム塩などのジアルキルアンモニウム塩、トリメチルアンモニウム塩またはトリエチルアンモニウム塩などのトリアルキルアンモニウム塩が挙げられる。
【００１１】
本願発明の別の側面によれば、炎症関連疾患の治療または予防のための医薬品組成物であって、治療有効量の上記式（１）で表される化合物、または薬学的に許容されうるその塩を含む、前記医薬品組成物が提供される。
【００１２】
ここで、炎症関連疾患とは、アレルギー性疾患、膠原病等が挙げられ、アレルギー性疾患としては、アトピー性皮膚炎、花粉症、接触性皮膚炎、気管支炎などが挙げられる。
【００１３】
本願発明のさらに別の側面によれば、上記式（１）で表される化合物、または食品として許容されうるその塩を添加物として含む、食品が提供される。
ここで、食品としては、特に限定されるものではないが、アレルギー予防効果を有する食品、保健機能食品、健康食品および一般食品が挙げられ、上記化合物を含む以外のアレルギーを予防するための処理が施されていてもよい。
【００１４】
【発明の実施の形態】
以下、本発明を更に具体的に説明する。
本発明の抗アレルギー剤は、ヒスタミンの遊離抑制作用を有するため、Ｉ型アレルギーに関連するアレルギー性疾患を効果的に予防および治療することができ、さらに、ＩＶ型アレルギーの予防および治療にも効果を示すことから、アトピー性皮膚炎に対する有効な予防薬および治療薬となりうるものである。従って、本発明の抗アレルギー剤は、医薬組成物の有効成分として使用することができる。当該医薬組成物は、経口投与剤および非経口投与剤のいずれであってもよい。経口投与の場合は、軟・硬カプセル剤または、錠剤、顆粒剤、細粒剤、散剤として投与することができる。非経口投与の方法としては、局所組織内投与、皮内、皮下、筋肉内および静脈内注射などの他、局所への塗布、噴霧などが挙げられる。本発明の医薬組成物は、一般に用いられる各種成分を含みうるものであり、例えば、１種もしくはそれ以上の薬学的に許容され得る賦形剤、希釈剤、湿潤剤、乳化剤、分散剤、補助剤、防腐剤、緩衝剤、結合剤、安定剤等を含みうる。また本発明の抗アレルギー剤を局所に塗布する際の製剤の形態は、特に限定されないが、例えば、軟膏、クリームローションおよびスプレーなどとすることができる。
【００１５】
本発明の抗アレルギー剤の投与量は、患者の体型、年齢、体調、疾患の度合い、発症後の経過時間等により、適宜選択することができるが、例えば、一般に１〜５０００ｍｇ／日／成人の用量で使用され、好ましくは１〜５００ｍｇ／日／成人の用量で使用される。
【００１６】
また、本発明の抗アレルギー剤は、医薬品として用いるほか、医薬部外品、化粧品、食品等の成分、食品添加物などとして使用することができる。当該使用により、本発明の抗アレルギー剤の日常的および継続的な摂取が可能となり、効果的なアレルギー体質の改善、およびアレルギーの発症の予防が可能となる。本発明の抗アレルギー剤が食品素材として使用される例としては、アレルギー予防効果を有する機能性食品、健康食品、一般食品（ジュース、菓子、加工食品等）、栄養補助食品（栄養ドリンク等）が挙げられる。
【００１７】
本発明の抗アレルギー物質を化粧品等の成分とする場合は、特に限定されるものではないが、化粧水、乳液、石鹸、洗顔料、シャンプーおよびリンスなどの成分とすることができる。
【００１８】
【実施例】
以下、本発明の好適な実施例についてさらに詳細に説明するが、本発明はこれらの実施例に限定されるものではない。
【００１９】
【実施例１】ヒスタミン遊離抑制活性測定
１） 細胞および試薬
本測定においては、肥満細胞系列のラット好塩基球白血病細胞ＲＢＬ−２Ｈ３細胞を用いた。当該細胞は、東北大学加齢医学研究所より供与された。ヒスタミン遊離刺激物質として用いたコンカナバリンＡ（Ｃｏｎ Ａ）およびカルシウムイオノフォア Ａ２３１８７は、それぞれシグマ‐アルドリッチ ジャパン（株）から入手した。１０％ＦＢＳを含むＲＰＭＩ培地は、シグマ‐アルドリッチ ジャパン（株）から入手した。試験薬物として用いたセサモールは東京化成工業（株）より入手し、メトキシヒドロキシキノン、フェルラ酸およびカフェー酸は和光純薬工業（株）より入手した。ポストラベル化法に用いた高速液体クロマトグラフィー（ＨＰＬＣ）システムは、以下の通りであった。
【００２０】
分離カラム： Ｌ−ｃｏｌｕｍｎ ＯＤＳ（２５０×４．６ｍｍ、化学物質評価研究機構）
プレカラム： μＢｏｎｄａｐａｋ Ｃ１８ Ｇｕａｒｄ−Ｐａｋ（Ｗａｔｅｒｓ）
移動層： Ａ液：Ｂ液＝２：１
Ａ液： １００ｍＭ 酒石酸ナトリウムバッファー − １０ｍＭ １−オクタンスルホン酸ナトリウム（ｐＨ４．４）
Ｂ液： メタノール
流速： １．０ｍＬ／分、 温度： ５０℃
反応コイル： ステンレススチールチュービング（４ｍ×０．５ｍｍ）
移動層： Ａ液：Ｂ液＝２：１
Ａ液： ４００ｍＭ ホウ酸ナトリウムバッファー（ｐＨ９．２）
Ｂ液： １０ｍＭ ｏ−フタルアルデヒド メタノール溶液
流速： ０．５ｍＬ／分、 温度： ５０℃
ポンプ： 島津ＨＰＬＣ用送液ユニットＬＣ−６Ａ
検出器： 島津分光蛍光ＨＰＬＣモニタＲＦ−５３５（Ｒａｎｇｅ：１６、Ｓｅｎｓｉｔｉｖｉｔｙ：ｈｉｇｈ、Ｒｅｓｐｏｎｓｅ：ｍｅｄｉｕｍ）、励起：３６０ｎｍ、放射：４４０ｎｍ
データ処理装置：島津クロマトパックＣ−Ｒ７Ａ ｐｌｕｓ（ＡＴＴＥＮ４）
２） ＲＢＬ−２Ｈ３細胞の培養
１０％ＦＢＳ含有ＲＰＭＩ−１６４０培地を用いて、３７℃、ＣＯ_２インキュベーターで常法により培養した。試験に用いる際は、トリプシン処理後、１×１０^５個／ｍＬの細胞濃度になるように０．０５％ＢＳＡ−Ｔｙｒｏｄｅバッファーを加え（４℃）、４００μＬの細胞懸濁液として使用した。
【００２１】
３） 測定方法
４００μＬの細胞懸濁液に試験検体溶液または懸濁液（ＰＢＳまたは４％ＤＭＳＯ−ＰＢＳ）５０μＬを加え（最終濃度として５０μｇ／ｍＬ）、１０分間３７℃でプレインキュベーションした。ヒスタミン遊離刺激物質として、コカナバリンＡ（Ｃｏｎ Ａ）の生理食塩水溶液２５μＬおよびホスファチジルセリンのＰＢＳ乳化液（最終濃度として、それぞれ１００μｇ／ｍＬ、２５μｇ／ｍＬ）、またはＣａイオノフォアＡ２３１８７の１０％ＤＭＳＯ−ＰＢＳ溶液の懸濁液５０μＬ（最終濃度として１μｇ／ｍＬ）を加え、６０分間３７℃でインキュベーションをした。その後、氷冷することにより反応を停止し、遠心分離により（１０００ｒｐｍ、５分間）上清と沈渣を分取した。
【００２２】
得られた上清２００μＬに、１６０μＬのＰＢＳおよび４０μＬのスペルミジン水溶液（０．５ｍｇ／ｍＬ）を加え、５０μＬを上述のＨＰＬＣシステムに注入し、ポストラベル化法によりヒスタミンを定量した。
【００２３】
得られた沈渣に、２００μＬのＢＳＡ（０．０５％）−Ｔｙｒｏｄｅバッファー、１６０μＬの０．１Ｎ−ＨＣｌおよび４０μＬのスペルミジン水溶液（０．５ｍｇ／ｍＬ）を加え、５０μＬを上述のＨＰＬＣシステムに注入し、ポストラベル化法によりヒスタミンを定量した。
【００２４】
上清中のヒスタミン量と沈渣中のヒスタミン量をあわせた量を全ヒスタミン量とし、この全ヒスタミン量に対する上清中のヒスタミン量の割合をヒスタミン遊離率とした。各試験検体の抑制効果は、試験検体を加えなかった場合のヒスタミン遊離の量を１００％として、ヒスタミン遊離の抑制率として算出した。
。結果を表１に示す。
【００２５】
【表１】

【００２６】
【実施例２】ＩＶ型アレルギーに対する効果の測定
１） 動物および試薬
日本エスエルシー（株）より購入し１週間飼育環境に馴化させた６週齢のｄｄＹ系雄性マウスを用いた。抗原として用いた４−エトキシメチレン−２−フェニルオキサゾール−５−オン（以下、オキサゾロンとする）は、シグマ‐アルドリッチ ジャパン（株）より入手し、アセトン：オリーブオイル＝４：１溶液に溶解して使用した。セサモールは東京化成工業（株）より入手し、蒸留水に溶解して使用した。プレドニゾロンはシグマ‐アルドリッチ ジャパン（株）より入手し、オリーブオイルに懸濁させた状態で使用した。
【００２７】
２） 感作マウスの作製および皮膚反応惹起
マウスの腹部の毛刈り後、オキサゾロンの２％（ｗ／ｖ）溶液を５０μＬずつ各個体の腹部に塗布し、一次感染させた。７日後、各固体の右耳の表裏両側にオキサゾロンの１％（ｗ／ｖ）溶液を５０μＬずつ塗布し２次感作させ皮膚反応を惹起させた。コントロール群にはオキサゾロンの溶解に用いたｖｅｈｉｃｌｅ（アセトン：オリーブオイル＝４：１溶液）を腹部に５０μＬずつ腹部に塗布した。
【００２８】
３） 薬物の投与
１次感作させたマウスを５群（各群につき検体数４）に分割し、セサモールは、摂食による吸収低下を避けるため３時間の絶食を行った後、５群のうちの３群に対して２次感作の１時間前と６時間後に、それぞれ７０ｍｇ／ｋｇ、１４０ｍｇ／ｋｇおよび２８０ｍｇ／ｋｇを経口投与（ｐ．ｏ．）した。残りの２群のうちの１群に対して、プレドニゾロン（ＰＤＺ）の２．５ｍｇ／ｋｇを経口投与した。残りの１群はコントロール群とし、ｖｅｈｉｃｌｅ（オリーブオイル）を経口投与した。２次感作２４時間後に耳の厚さを測定し、反応前との差を取り、腫脹とした。結果を図１に示す。
【００２９】
図１の結果は、セサモールはオキサゾールにより誘発された耳介腫脹を用量依存的に抑制することを示すものである。
【００３０】
【実施例３】三相性皮膚反応に対する効果の測定
免疫グロブリンＥ（ＩｇＥ）抗体をマウスに受動感作させ、抗原を耳介に塗布することに起因する耳介腫脹において、塗布後約１時間後に第１相（ｉｍｍｅｄｉａｔｅ ｐｈａｓｅ ｒｅｓｐｏｎｓｅ：ＩＰＲ）の、約２４時間後に第２相（ｌａｔｅ ｐｈａｓｅ ｒｅｓｐｏｎｓｅ：ＬＰＲ）の、および塗布後約７〜８日後に第３相（ｖｅｒｙ ｌａｔｅ ｐｈａｓｅ ｒｅｓｐｏｎｓｅ：ｖＬＰＲ）のピークが観察される（Ａｌｌｅｒｇｏｌｏｇｙ Ｉｎｔｅｒｎａｔｉｏｎａｌ ４８： ２６５−２７３ （１９９９）、およびＢｉｏｌ． Ｐｈａｒｍ． Ｂｕｌｌ． １８ （２）： ２３９−２４５ （１９９５）を参照のこと）。本実験項では、第１−３相の各々におけるセサモールの効果について説明する。
【００３１】
１） 動物および試薬
日本エスエルシー（株）より購入し１週間飼育環境に馴化させた６週齢のＢＡＬＢ／ｃ系雄性マウスを用いた。抗ジニトロフェノール（ＤＮＰ）マウスＩｇＥ抗体（ＰＣＡ ｔｉｔｅｒ×１０００００）はシグマ‐アルドリッチ ジャパン（株）より入手し、生理食塩水による１０倍希釈溶液として使用した。ジニトロフルオロベンゼン（ＤＮＦＢ）はナカライテスク（株）より入手し、０．１％（ｗ／ｖ）溶液（アセトン：オリーブオイル＝３：１溶液）として使用した。セサモールは東京化成工業（株）より入手し、蒸留水に溶解して使用した。
【００３２】
２） 受動感作マウスの作製および皮膚反応惹起
マウスの尾静脈に０．１ｍＬ／個体の量の抗ＤＮＰマウスＩｇＥ抗体０．１ｍｇ／ｍＬの溶液を投与し、受動感作マウスを作成した。感作２４時間後に右耳の表裏両側に０．１％（ｗ／ｖ）のＤＮＦＢ溶液を１０μＬずつ塗布し、三相性皮膚反応を惹起させた。コントロール群にはＤＮＦＢの溶解に用いたｖｅｈｉｃｌｅ（アセトン：オリーブオイル＝３：１溶液）を同様に１０μＬずつ右耳に塗布した。
【００３３】
３） 薬物の投与
受動感作させたマウスを３群（各群につき検体数７）に分割し、セサモールは、摂食による吸収低下を避けるため３時間の絶食を行った後、３群のうちの１群に対して反応惹起の１時間前に２８０ｍｇ／ｋｇを経口投与した。残りの２群のうちの１群に対して、反応惹起の５時間後にセサモール２８０ｍｇ／ｋｇを経口投与した。残りの１群はコントロール群とし、ｖｅｈｉｃｌｅ（蒸留水）を投与した。反応惹起から一定時間後（１、４、２４、４８、７２、以降２４時間毎）に耳の厚さを測定し、反応前との差を取り、腫脹とした。結果を図２に示す。 図２に示した結果から、反応１時間前にセサモールを投与した群においては第１相反応（ＩＰＲ）における抑制効果が、反応５時間後にセサモールを投与した群においては第２相反応（ＬＰＲ）および第３相反応（ｖＬＰＲ）における抑制効果が確認された。
【００３４】
【実施例４】第３相反応（ｖＬＰＲ）に対する効果の測定
実施例３と同様の手順で、受動感作マウスの作成および皮膚反応惹起を行った。
【００３５】
受動感作させたマウスを３群（各群につき検体数７）に分割し、セサモールは、摂食による吸収低下を避けるため３時間の絶食を行った後、３群のうちの１群に対して反応惹起後２日目より７日目まで毎日１日２回、セサモール１４０ｍｇ／ｋｇ（１回の投与量）を経口投与した。残りの２群のうちの１群に対して、反応惹起後２日目より７日目まで毎日１日２回、クルクミン１５０ｍｇ／ｋｇ（１回の投与量）を経口投与した。残りの１群はコントロール群とし、ｖｅｈｉｃｌｅ（蒸留水）を同様に投与した。反応惹起から一定時間後（１、４、２４、４８、７２、以降２４時間毎）に耳の厚さを測定し、反応前との差を取り、腫脹とした。結果を図３に示す。図３は各群の耳介腫脹の経時変化を、図４は反応惹起から１６８時間後の耳介腫脹を示すものである。
【００３６】
図３および図４の結果は、セサモールは第３相反応（ｖＬＰＲ）において耳介腫脹を有意に抑制することを示すものである。
【００３７】
【実施例５】ＩＬ−１β投与により誘起される耳介腫脹に対するセサモールの効果の測定
１） 動物および試薬
日本エスエルシー（株）より購入し１週間飼育環境に馴化させた６週齢のＢＡＬＢ／ｃ系雄性マウスを用いた。ＩＬ−１βはペプロテック（株）より入手し、ＩＬ−１βの１％（ｗ／ｖ）水溶液として使用した。セサモールは東京化成工業（株）より入手し、蒸留水に溶解して使用した。アミノグアニジンクロリドは和光純薬工業（株）より入手し、生理食塩水に溶解して使用した。
【００３８】
２） 皮膚反応惹起
マウスの耳介に１％（ｗ／ｖ）のＩＬ−１β溶液の１５μＬを皮内投与することにより皮膚反応を惹起した。
【００３９】
３） 薬物の投与
ＩＬ−１βを投与したマウスを３群（各群につき検体数７）に分割し、セサモールは、摂食による吸収低下を避けるため３時間の絶食を行った後、３群のうちの１群に対して投与の１時間前および７時間後に１４０ｍｇ／ｋｇを経口投与した。残りの２群のうちの１群に対して、投与の１時間前および７時間後に３０ｍｇ／ｋｇのアミノグアニジンクロリド（ＡＧ）を経口投与した。残りの１群はコントロール群とし、 ｖｅｈｉｃｌｅ（蒸留水）を経口投与した。投与１時間前および投与から一定時間後（１、４、８、１２、２４および４８時間後）に耳の厚さを測定し、反応前との差を取り、腫脹とした。結果を図５に示す。
【００４０】
図５の結果は、セサモールがＩＬ−１βの皮内投与による耳介腫脹に対して有意な抑制効果を有することを示している。
【００４１】
【発明の効果】
本発明の抗アレルギー剤は、ヒスタミン遊離抑制作用を有し、さらに三相性皮膚反応における第３層反応（ｖＬＰＲ）に対して抑制効果を有することから、本発明は、アレルギー性疾患、特にアトピー性皮膚炎に対する有効な治療手段を提供する。
【図面の簡単な説明】
【図１】図１は、本発明の抗アレルギー剤が、オキサゾールにより誘発された耳介腫脹を用量依存的に抑制することを示すものである。
【図２】図２は、本発明の抗アレルギー剤を、反応１時間前に投与した群においては第１相反応（ＩＰＲ）における抑制効果が、反応５時間後に投与した群においては第２相反応（ＬＰＲ）および第３相反応（ｖＬＰＲ）における抑制効果が確認されたことを示すものである。
【図３】図３は、実施例４の検体の各群の耳介腫脹を経時的に示すものであり、本発明の抗アレルギー剤が、第３相反応（ｖＬＰＲ）において耳介腫脹を有意に抑制することを示すものである。
【図４】図４は、実施例４の検体の各群の耳介腫脹について、反応惹起後１６８時間後の値を示すものである。
【図５】図５は、本発明の抗アレルギー剤がＩＬ−１βの皮内投与による耳介腫脹に対して有意な抑制効果を有することを示すものである。[0001]
[Industrial applications]
The present application relates to pharmaceuticals, quasi-drugs, foods, cosmetics, etc., containing phenolic compounds having pharmacological effects and used for the prevention or treatment of allergic inflammatory-related diseases such as atopic dermatitis.
[0002]
[Prior art]
Allergic diseases such as hay fever and atopic dermatitis have recently increased in the number of patients. Allergies are classified into four types (types I to IV) according to the mechanism of onset. Type I allergy is related to hay fever and the like, type IV allergy is related to contact dermatitis and the like. It is thought that both type I allergy as an immediate reaction and type IV allergy as a delayed reaction are deeply involved in dermatitis. Type I allergic reactions are caused by chemical mediators such as histamine released from mast cells sensitized by immunoglobulin E antibody (IgE). Type IV allergy is caused by cytokines released by T cells sensitized by macrophages.
[0003]
On the other hand, some reports have already been made on the physiological activity of phenolic compounds derived from natural products. Sesamol, ellagic acid, gallic acid, ferulic acid, dl-tocopherol are known to be antioxidants (non-oxidizing substances). Patent Document 1), and gallic acid derivatives that affect cytokine secretion have also been reported (see Non-Patent Documents 2 and 3). It is also known that sesamol has a protective effect against drug-induced liver injury (see Non-Patent Document 4).
[0004]
[Non-patent document 1]
Pharmaceutical Magazine, Vol. 110 (No. 9), pp. 697-701, 1990 [Non-Patent Document 2]
Pharmaceutical Magazine, Vol. 120 (No. 6), pp. 408-412, 2000 [Non-Patent Document 3]
Pharmaceutical Magazine, Vol. 121 (No. 6), pp. 451-457, 2001 [Non-Patent Document 4]
Pharmaceutical Magazine, Vol. 114 (No. 11), pp. 901-910, 1994
[Problems to be solved by the invention]
Allergic diseases, especially atopic dermatitis, often have remarkable external symptoms, and the number of patients has been increasing in recent years. Therefore, an effective treatment method is required.
[0006]
In addition, steroidal antiallergic agents that are effective for both type I allergy and type IV allergy having different onset mechanisms may have side effects, so that their use may be limited. There is a demand for the development of an antiallergic agent that is effective for both thyroid and type IV allergy.
[0007]
[Means for Solving the Problems]
The present inventor has conducted intensive studies with a focus on food-derived natural products in order to solve such problems, and found that a specific phenolic compound has an antiallergic effect.
[0008]
An object of the present invention is to provide an antiallergic agent derived from a natural product or a derivative thereof, which can effectively prevent and treat allergic diseases.
That is, according to one aspect of the present invention, equation (1):
[0009]
Embedded image

[0010]
(Wherein, R ¹ and R ² are each independently a hydrogen atom, a methyl group, an ethyl group, or an isopropyl group, and R ¹ and R ² are taken together to form —CH ₂ — Well,
R ³ is a hydroxyl group, —CH ＝ CH— (CH ₂ ) _n —COOH, or —O-sugar;
n is an integer from 0 to 5)
Or an antiallergic agent comprising a pharmaceutically acceptable salt thereof.
Here, "sugar" is a general sugar present in a living body, and may be a monosaccharide or 2 to 10 oligosaccharides. For example, hexasaccharide such as glucose, rhamnose, and galactose, and fructose And saccharides composed of pentasaccharides. Examples of the compound represented by the formula (1) include sesamol, caffeic acid, and a derivative thereof as a prodrug. The pharmaceutically acceptable salts thereof are not particularly limited, and include, for example, metal salts such as sodium salt, potassium salt, magnesium salt and calcium salt, ammonium salts, methylammonium salts and ethylammonium salts and the like. Examples include dialkylammonium salts such as monoalkylammonium salts, dimethylammonium salts or diethylammonium salts, and trialkylammonium salts such as trimethylammonium salts or triethylammonium salts.
[0011]
According to another aspect of the present invention, there is provided a pharmaceutical composition for treating or preventing an inflammatory-related disease, comprising a therapeutically effective amount of a compound represented by the above formula (1), or a pharmaceutically acceptable salt thereof. A pharmaceutical composition is provided, comprising a salt.
[0012]
Here, the inflammation-related diseases include allergic diseases, collagen diseases and the like, and the allergic diseases include atopic dermatitis, hay fever, contact dermatitis, bronchitis and the like.
[0013]
According to still another aspect of the present invention, there is provided a food comprising, as an additive, the compound represented by the above formula (1) or a salt thereof which is acceptable as a food.
Here, examples of the food include, but are not particularly limited to, foods having an allergy prevention effect, health functional foods, health foods, and general foods, and a treatment for preventing allergies other than the above-mentioned compounds is performed. It may be applied.
[0014]
BEST MODE FOR CARRYING OUT THE INVENTION
Hereinafter, the present invention will be described more specifically.
Since the antiallergic agent of the present invention has a histamine release inhibitory effect, it can effectively prevent and treat allergic diseases related to type I allergy, and is also effective in preventing and treating type IV allergy. Therefore, it can be an effective prophylactic and therapeutic agent for atopic dermatitis. Therefore, the antiallergic agent of the present invention can be used as an active ingredient of a pharmaceutical composition. The pharmaceutical composition may be any of an orally administered drug and a parenterally administered drug. In the case of oral administration, it can be administered as soft or hard capsules or tablets, granules, fine granules, and powders. Examples of the method of parenteral administration include local application into a tissue, intradermal, subcutaneous, intramuscular, and intravenous injection, as well as topical application and spraying. The pharmaceutical composition of the present invention may contain various commonly used components, for example, one or more pharmaceutically acceptable excipients, diluents, humectants, emulsifiers, dispersants, and auxiliaries. Agents, preservatives, buffers, binders, stabilizers and the like. The form of the preparation when the antiallergic agent of the present invention is applied topically is not particularly limited, and may be, for example, an ointment, a cream lotion, a spray or the like.
[0015]
The dose of the antiallergic agent of the present invention can be appropriately selected depending on the patient's body type, age, physical condition, degree of disease, elapsed time after onset, and the like. For example, generally, the dose is 1 to 5000 mg / day / adult. It is used in doses, preferably in doses of 1 to 500 mg / day / adult.
[0016]
The antiallergic agent of the present invention can be used not only as a pharmaceutical, but also as a quasi-drug, a cosmetic, a component of food and the like, a food additive and the like. By the use, daily and continuous ingestion of the antiallergic agent of the present invention becomes possible, and it becomes possible to effectively improve the allergy constitution and prevent the onset of allergy. Examples of the use of the antiallergic agent of the present invention as a food material include functional foods having a preventive effect on allergies, health foods, general foods (juices, confectionery, processed foods, etc.), and dietary supplements (nutrition drinks, etc.). No.
[0017]
When the antiallergic substance of the present invention is used as a component of cosmetics or the like, it is not particularly limited, but may be a component such as lotion, emulsion, soap, face wash, shampoo, and rinse.
[0018]
【Example】
Hereinafter, preferred embodiments of the present invention will be described in more detail, but the present invention is not limited to these embodiments.
[0019]
Example 1 Measurement of Histamine Release Inhibitory Activity 1) Cells and Reagents In this measurement, mast cell line rat basophil leukemia cells RBL-2H3 cells were used. The cells were provided by Tohoku University Institute for Aging and Medicine. Concanavalin A (Con A) and calcium ionophore A23187 used as histamine release stimulating substances were obtained from Sigma-Aldrich Japan Co., Ltd., respectively. RPMI medium containing 10% FBS was obtained from Sigma-Aldrich Japan. Sesamol used as a test drug was obtained from Tokyo Chemical Industry Co., Ltd., and methoxyhydroxyquinone, ferulic acid and caffeic acid were obtained from Wako Pure Chemical Industries, Ltd. The high performance liquid chromatography (HPLC) system used for the post-labeling method was as follows.
[0020]
Separation column: L-column ODS (250 x 4.6 mm, Chemical Substance Evaluation Research Organization)
Pre-column: μBondapak C18 Guard-Pak (Waters)
Moving bed: Liquid A: Liquid B = 2: 1
Solution A: 100 mM sodium tartrate buffer-10 mM sodium 1-octanesulfonate (pH 4.4)
Liquid B: methanol flow rate: 1.0 mL / min, temperature: 50 ° C.
Reaction coil: Stainless steel tubing (4m × 0.5mm)
Moving bed: Liquid A: Liquid B = 2: 1
Solution A: 400 mM sodium borate buffer (pH 9.2)
Solution B: 10 mM o-phthalaldehyde methanol solution flow rate: 0.5 mL / min, temperature: 50 ° C.
Pump: Shimadzu HPLC liquid sending unit LC-6A
Detector: Shimadzu Spectrofluorimetric HPLC Monitor RF-535 (Range: 16, Sensitivity: high, Response: medium), Excitation: 360 nm, Emission: 440 nm
Data processing device: Shimadzu Chromatopack C-R7A plus (ATTEN4)
2) Culture of RBL-2H3 cells Using RPMI-1640 medium containing 10% FBS, the cells were cultured in a CO ₂ incubator at 37 ° C by a conventional method. When used for the test, 0.05% BSA-Tyrode buffer was added (4 ° C.) to a cell concentration of 1 × 10 ⁵ cells / mL after trypsin treatment, and used as a 400 μL cell suspension.
[0021]
3) Measurement method 50 μL of a test sample solution or suspension (PBS or 4% DMSO-PBS) was added to 400 μL of the cell suspension (final concentration: 50 μg / mL), and preincubation was performed at 37 ° C. for 10 minutes. As a histamine release stimulating substance, 25 μL of a physiological saline solution of cocanavalin A (Con A) and a PBS emulsion of phosphatidylserine (final concentrations of 100 μg / mL and 25 μg / mL, respectively), or a 10% DMSO-PBS solution of Ca ionophore A23187 Was added (final concentration: 1 μg / mL), and the mixture was incubated at 37 ° C. for 60 minutes. Thereafter, the reaction was stopped by cooling with ice, and the supernatant and sediment were separated by centrifugation (1000 rpm, 5 minutes).
[0022]
To 200 μL of the obtained supernatant, 160 μL of PBS and 40 μL of an aqueous solution of spermidine (0.5 mg / mL) were added, 50 μL was injected into the above-mentioned HPLC system, and histamine was quantified by a post-labeling method.
[0023]
To the obtained precipitate, 200 μL of BSA (0.05%)-Tyrode buffer, 160 μL of 0.1 N HCl and 40 μL of an aqueous spermidine solution (0.5 mg / mL) were added, and 50 μL was injected into the above-mentioned HPLC system. Histamine was quantified by a post-labeling method.
[0024]
The sum of the amount of histamine in the supernatant and the amount of histamine in the sediment was defined as the total histamine amount, and the ratio of the amount of histamine in the supernatant to the total histamine amount was defined as the histamine release rate. The inhibitory effect of each test sample was calculated as the histamine release inhibitory rate, with the amount of histamine release when no test sample was added as 100%.
. Table 1 shows the results.
[0025]
[Table 1]

[0026]
Example 2 Measurement of Effect on Type IV Allergy 1) Animals and Reagents Six-week-old male ddY mice, purchased from SLC Japan Ltd. and adapted to a breeding environment for one week, were used. 4-ethoxymethylene-2-phenyloxazol-5-one (hereinafter referred to as oxazolone) used as the antigen was obtained from Sigma-Aldrich Japan Co., Ltd., and dissolved in a 4: 1 acetone: olive oil solution. used. Sesamol was obtained from Tokyo Chemical Industry Co., Ltd. and used after dissolving in distilled water. Prednisolone was obtained from Sigma-Aldrich Japan Co., Ltd. and used in a state of being suspended in olive oil.
[0027]
2) Preparation of sensitized mice and skin reaction induced mice. After shaving the abdomen of the mice, 50 μL of a 2% (w / v) solution of oxazolone was applied to the abdomen of each individual to carry out primary infection. Seven days later, 50 μL of a 1% (w / v) solution of oxazolone was applied to both front and back of the right ear of each solid, and secondary sensitization was performed to induce a skin reaction. To the control group, 50 μL of the vehicle (acetone: olive oil = 4: 1 solution) used for dissolving oxazolone was applied to the abdomen by 50 μL.
[0028]
3) Administration of drug The mice sensitized to primary sensitization were divided into 5 groups (4 specimens per group), and sesamol was fasted for 3 hours to avoid a decrease in absorption due to feeding. One hour before and six hours after the second sensitization, 70 mg / kg, 140 mg / kg and 280 mg / kg were orally administered (po) to three of the three groups. To one of the remaining two groups, 2.5 mg / kg of prednisolone (PDZ) was orally administered. The remaining one group was a control group, and vehicle (olive oil) was orally administered. Twenty-four hours after the second sensitization, the thickness of the ear was measured, and the difference from the value before the reaction was determined to be swelling. The results are shown in FIG.
[0029]
The results in FIG. 1 show that sesamol dose-dependently suppresses oxazole-induced auricular swelling.
[0030]
Example 3 Measurement of Effect on Triphasic Skin Reaction In an ear swelling caused by passively sensitizing a mouse with an immunoglobulin E (IgE) antibody and applying the antigen to the auricle, about 1 hour after application Peaks of the first phase (immediate phase response: IPR), about 24 hours after the second phase (late phase response: LPR), and about 7 to 8 days after application, the peak of the third phase (very late phase response: vLPR). (See Allergy International 48: 265-273 (1999) and Biol. Pharm. Bull. 18 (2): 239-245 (1995)). In this experimental section, the effect of sesamol in each of the first to third phases will be described.
[0031]
1) Animals and Reagents Six-week-old BALB / c male mice purchased from Japan SLC, Inc. and adapted to a breeding environment for one week were used. Anti-dinitrophenol (DNP) mouse IgE antibody (PCA titer × 100,000) was obtained from Sigma-Aldrich Japan Co., Ltd. and used as a 10-fold diluted solution with physiological saline. Dinitrofluorobenzene (DNFB) was obtained from Nacalai Tesque, Inc. and used as a 0.1% (w / v) solution (acetone: olive oil = 3: 1 solution). Sesamol was obtained from Tokyo Chemical Industry Co., Ltd. and used after dissolving in distilled water.
[0032]
2) Production of passively sensitized mice and skin reaction-induced mice were administered with 0.1 mL / individual solution of 0.1 mg / mL anti-DNP mouse IgE antibody in the tail vein of mice to produce passively sensitized mice. Twenty-four hours after sensitization, a 0.1% (w / v) DNFB solution was applied to each of the front and back sides of the right ear in an amount of 10 μL each to induce a three-phase skin reaction. To the control group, 10 μL of the vehicle (acetone: olive oil = 3: 1 solution) used for dissolving DNFB was similarly applied to the right ear by 10 μL.
[0033]
3) Administration of the drug The passively sensitized mice were divided into three groups (seven samples per group), and sesamol was fasted for 3 hours to avoid a decrease in absorption due to feeding. One group was orally administered 280 mg / kg one hour before the induction of the reaction. To one of the remaining two groups, sesamol 280 mg / kg was orally administered 5 hours after the induction of the reaction. The remaining one group was a control group, and was administered with a vehicle (distilled water). After a certain time (1, 4, 24, 48, 72, and thereafter every 24 hours) from the induction of the reaction, the thickness of the ear was measured, and the difference from the thickness before the reaction was determined to be swelling. FIG. 2 shows the results. From the results shown in FIG. 2, the inhibitory effect on the first phase reaction (IPR) in the group to which sesamol was administered 1 hour before the reaction was observed, and the second phase reaction (LPR) in the group to which sesamol was administered 5 hours after the reaction. And the inhibitory effect on the third phase reaction (vLPR) was confirmed.
[0034]
Example 4 Measurement of Effect on Phase 3 Response (vLPR) Passive sensitized mice were prepared and skin reactions were induced in the same manner as in Example 3.
[0035]
The passively sensitized mice were divided into 3 groups (7 specimens per group), and sesamol was fasted for 3 hours to avoid a decrease in absorption due to feeding. Then, sesamol 140 mg / kg (single dose) was orally administered twice a day from day 2 to day 7 after the reaction was induced. One of the remaining two groups was orally administered curcumin 150 mg / kg (single dose) twice a day from day 2 to day 7 after the reaction was induced. The other group was a control group, and vehicle (distilled water) was similarly administered. After a certain time (1, 4, 24, 48, 72, and thereafter every 24 hours) from the induction of the reaction, the thickness of the ear was measured, and the difference from the thickness before the reaction was determined to be swelling. The results are shown in FIG. FIG. 3 shows the time course of pinna swelling in each group, and FIG. 4 shows pinna swelling 168 hours after the induction of the reaction.
[0036]
The results in FIGS. 3 and 4 show that sesamol significantly suppresses pinna swelling in the phase 3 reaction (vLPR).
[0037]
Example 5 Measurement of the Effect of Sesamol on Auricular Swelling Induced by IL-1β Administration 1) Animals and Reagents 6-week-old BALB / PB purchased from Japan SLC, Inc. and adapted to a 1-week rearing environment Male c-line mice were used. IL-1β was obtained from Peprotech Co., Ltd. and used as a 1% (w / v) aqueous solution of IL-1β. Sesamol was obtained from Tokyo Chemical Industry Co., Ltd. and used after dissolving in distilled water. Aminoguanidine chloride was obtained from Wako Pure Chemical Industries, Ltd. and used after dissolving in physiological saline.
[0038]
2) Induction of skin reaction A skin reaction was induced by intradermally administering 15 μL of a 1% (w / v) IL-1β solution to the auricle of a mouse.
[0039]
3) Administration of Drug Mice administered IL-1β were divided into three groups (7 specimens per group), and sesamol was fasted for 3 hours to avoid a decrease in absorption due to feeding. One group was orally administered 140 mg / kg 1 hour before and 7 hours after administration. One of the remaining two groups was orally administered 30 mg / kg of aminoguanidine chloride (AG) 1 hour before and 7 hours after administration. The remaining one group was a control group, and vehicle (distilled water) was orally administered. The thickness of the ear was measured 1 hour before administration and at a certain time after administration (1, 4, 8, 12, 24 and 48 hours). FIG. 5 shows the results.
[0040]
The results in FIG. 5 indicate that sesamol has a significant inhibitory effect on ear swelling due to intradermal administration of IL-1β.
[0041]
【The invention's effect】
Since the antiallergic agent of the present invention has an inhibitory effect on histamine release and further has an inhibitory effect on the third layer reaction (vLPR) in a triphasic skin reaction, the present invention provides an allergic disease, particularly an atopic disease. An effective treatment for dermatitis is provided.
[Brief description of the drawings]
FIG. 1 shows that the antiallergic agent of the present invention inhibits oxazole-induced auricular swelling in a dose-dependent manner.
FIG. 2 shows that the anti-allergic agent of the present invention was administered 1 hour before the reaction, the inhibitory effect on the first phase reaction (IPR) was obtained, and the anti-allergic agent of the present invention was administered 5 hours after the reaction. It shows that the inhibitory effects on the reaction (LPR) and the third phase reaction (vLPR) were confirmed.
FIG. 3 shows the ear swelling of each group of the sample of Example 4 over time, wherein the antiallergic agent of the present invention significantly increased the ear swelling in a phase 3 reaction (vLPR). It is shown that it suppresses.
FIG. 4 shows the values of 168 hours after the induction of the reaction for the pinna swelling of each group of the sample of Example 4.
FIG. 5 shows that the antiallergic agent of the present invention has a significant inhibitory effect on auricular swelling due to intradermal administration of IL-1β.

Claims (11)

Equation (1):

(Wherein, R ¹ and R ² are each independently a hydrogen atom, a methyl group, an ethyl group, or an isopropyl group, and R ¹ and R ² are taken together to form —CH ₂ — Well,
R ³ is a hydroxyl group, —CH ＝ CH— (CH ₂ ) _n —COOH, or —O-sugar;
n is an integer from 0 to 5)
Or an antiallergic agent comprising a pharmaceutically acceptable salt thereof. Antiallergic agents containing sesamol or caffeic acid. Antiallergic agents, including sesamol. A pharmaceutical composition for treating or preventing inflammation-related diseases, comprising a therapeutically effective amount of formula (1):

(Wherein, R ¹ and R ² are each independently a hydrogen atom, a methyl group, an ethyl group, or an isopropyl group, and R ¹ and R ² are taken together to form —CH ₂ — Well,
R ³ is a hydroxyl group, —CH ＝ CH— (CH ₂ ) _n —COOH, or —O-sugar;
n is an integer from 0 to 5)
Or a pharmaceutically acceptable salt thereof. A pharmaceutical composition for treating or preventing an inflammatory-related disease, comprising a therapeutically effective amount of sesamol or caffeic acid. A pharmaceutical composition for treating or preventing an inflammatory-related disease, comprising a therapeutically effective amount of sesamol. The pharmaceutical composition according to any one of claims 4 to 6, wherein the inflammation-related disease is an allergic disease. The pharmaceutical composition according to claim 7, wherein the allergic disease is hay fever, atopic dermatitis, or contact dermatitis. Equation (1):

(Wherein, R ¹ and R ² are each independently a hydrogen atom, a methyl group, an ethyl group, or an isopropyl group, and R ¹ and R ² are taken together to form —CH ₂ — Well,
R ³ is a hydroxyl group, —CH ＝ CH— (CH ₂ ) _n —COOH, or —O-sugar;
n is an integer from 0 to 5)
Or a food additive having an allergy preventive effect, comprising a compound represented by the formula: or a salt thereof acceptable as a food. A food comprising the food additive according to claim 9. Equation (1):

(Wherein, R ¹ and R ² are each independently a hydrogen atom, a methyl group, an ethyl group, or an isopropyl group, and R ¹ and R ² are taken together to form —CH ₂ — Well,
R ³ is a hydroxyl group, —CH ＝ CH— (CH ₂ ) _n —COOH, or —O-sugar;
n is an integer from 0 to 5)
A cosmetic having an allergy preventive effect, comprising a compound represented by the formula: or a salt thereof acceptable as a food.

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