JP4072066B2 - Novel molecular recognition polymer and production method thereof - Google Patents

Description

【００３０】
架橋剤としては、付加重合可能な基を複数有するものが好ましい。付加重合可能な基を複数有する架橋剤の例としては、式（４）に示すｐ−ジビニルベンゼン、式（１８）に示すｏ−ジビニルベンゼン、式（１９）に示すｍ−ジビニルベンゼン等のジビニルベンゼン系架橋剤、（メタ）アクリル酸系架橋剤等がある。（メタ）アクリル酸系架橋剤としては、（メタ）アクリロイル基を複数有する化合物、例えば、式（１）に示すエチレングリコールジメタクリレート（ＥＤＭＡ）、式（２）に示すトリメチロールプロパントリメタクリレート（ＴＲＩＭ）、式（３）に示すトリメチロールプロパントリアクリレート、式（６）に示すテトラエチレングリコールジアクリレート、式（１１）に示すトリエチレングリコールジメタクリレート、式（１２）に示すペンタエリスリトールテトラアクリレート、式（１３）に示すペンタエリスリトールトリアクリレート、式（１４）に示すビスフェノールＡジメタクリレート、式（１５）に示すＮ，Ｎ’−メチレンジアクリルアミド、式（１６）に示すＮ，Ｎ’−１，４−フェニレンジアクリルアミド、式（１７）に示す３，５−ビス（アクリロイルアミド）安息香酸、式（２０）に示すＮ，Ｏ−ビスアクリロイル−Ｌ−フェニルアラニノール等が好適である。架橋剤として、ビニル基を１つだけ持つ化合物（式（５）に示す塩化ビニル、式（７）に示すスチレン、式（９）に示すフェニルビニルスルホキシド、式（１０）に示すフェニルビニルスルホン等）や、（メタ）アクリロイル基を１つだけ持つ化合物（例えば式（８）に示すメタクリル酸メチル等）等のような重合可能な基を１つだけ持つ化合物を用いてもよい。架橋剤としては、不飽和単量体が有する重合可能な基と同じ基を持つ架橋剤を使用することが好ましい。また、架橋剤は、結合部位同士のクロストークを避けるために不飽和単量体に対して大過剰で用いることが好ましい。具体的には、不飽和単量体１モルに対して架橋剤を５モル以上用いることが好ましい。
【００３１】
上記脱離工程では、重合工程で得られた重合体における官能基と標的分子との可逆的な結合を分解することにより、標的分子を脱離させる。例えば、可逆的な結合がエステル結合である場合には、エステル結合を酸加水分解あるいはアルカリ加水分解することにより標的分子を脱離させればよい。
【００３２】
〔非共有結合性分子認識ポリマーの製造方法〕
上記非共有結合性分子認識ポリマーは、いわゆる非共有結合型分子インプリンティングにより製造できる。すなわち、上記非共有結合性分子認識ポリマーは、上記非共有結合性官能基および光感応性部位を有する不飽和単量体を調製する調製工程と、上記不飽和単量体を、標的分子の存在下で架橋剤と共重合し、重合体を得る重合工程と、得られた重合体から標的分子を脱離させる脱離工程とを含む方法により製造できる。
【００３３】
上記不飽和単量体は、標的分子の持つ官能基と可逆的な非共有結合（水素結合など）を形成しうる官能基と、重合可能な不飽和結合と、光感応性部位とを持つ化合物である。標的分子の持つ官能基と可逆的な非共有結合を形成しうる官能基としては、標的分子の持つアミノ基と水素結合を形成しうるカルボニル基や、標的分子の持つ含窒素複素環の窒素原子と水素結合を形成しうる水酸基やアミノ基等が挙げられる。
【００３４】
上記調製工程では、例えば、光感応性部位を有する化合物にビニル基やメタクリロイル基等の付加重合可能な不飽和基をつけて付加重合可能とし、さらに標的分子と可逆的に非共有結合を形成しうる非共有結合性官能基（カルボキシル基、アミノ基、フェノール性水酸基等）をつけて付加重合可能な不飽和単量体を調製すればよい。
【００３５】
上記重合工程は、基本的には共有結合性分子認識ポリマーの製造方法と同様であるが、非共有結合型分子インプリンティングでは、標的分子と不飽和単量体との複合体を安定的に存在させるために、不飽和単量体は標的分子に対して過剰に用いることが好ましい。具体的には、標的分子１モルに対して標的分子を２モル以上用いることが好ましい。また、非共有結合が水素結合である場合、使用する溶媒としては、水素結合を妨害しないクロロホルム等のハロゲン系溶媒が好ましい。
【００３６】
上記脱離工程では、標的分子を選択的に溶解しうる溶液（例えばアルコール系溶媒等）で洗浄することにより、標的分子を脱離させる。これにより、非共有結合性分子認識ポリマーが得られる。
【００３７】
上記の非共有結合型分子インプリンティングにおいては、形成される結合部位の不均一性が問題となることがある。通常、標的分子と不飽和単量体（機能性モノマー）との複合体を形成させるために，不飽和単量体（機能性モノマー）は、標的分子に対して過剰に加えて重合する。当然ではあるが、結合部位に関与する不飽和単量体（機能性モノマー）は限られており、加えられた不飽和単量体（機能性モノマー）のうちの過剰分は、特異的な結合部位にはならず、非特異的な結合となる。すなわち、過剰分は、特異的結合部位と無関係に存在するか、あるいは完成度および特異性が低い結合部位として存在する。
【００３８】
そこで、上記の非共有結合型分子インプリンティングにおいては、この問題を解決する方法として、非共有結合型分子インプリンティングの後に、非共有結合型分子インプリンティングにより得られたインプリントポリマーの非特異的結合部位を化学修飾してブロックする処理（以下、「ポストインプリンティング処理」と称する）を行ってもよい。例えば、不飽和単量体（機能性モノマー）として不飽和カルボン酸を用いた非共有結合型分子インプリンティングによって得られたインプリントポリマーに対し、特異的結合部位を保護するために標的分子の共存下で、インプリントポリマー中における不飽和カルボン酸由来のカルボン酸残基で結合部位に関与していないものを、ヨウ化メチルによるメチルエステル化してもよい。このポストインプリンティング処理は、比較的簡便に、標的分子に対する非共有結合性分子認識ポリマーの選択性を向上させ、性能を改善できる可能性がある。
【００３９】
また、上記標的分子が水酸基を有し、かつ、上記非共有結合性分子認識ポリマーが標的分子の水酸基に対して特異的に可逆的な水素結合を形成する水酸基を有する場合には、共有結合型分子インプリンティングにより製造できる。すなわち、この場合、非共有結合性分子認識ポリマーは、上記光感応性部位および水酸基を有する不飽和単量体と、上記水酸基を有する標的分子との炭酸エステルを調製する調製工程と、上記炭酸エステルを架橋剤と共重合し、重合体を得る重合工程と、得られた重合体から、脱炭酸により標的分子を脱離させ、不飽和単量体由来の水酸基を遊離させる脱離工程とを含む方法で製造できる。
【００４０】
上記調製工程では、例えば、標的分子のクロロギ酸エステルを、トリエチルアミン存在下で光感応性部位および水酸基を有する不飽和単量体と反応させればよい。
【００４１】
上記重合工程は、共有結合性分子認識ポリマーの製造方法と同様である。上記脱離工程では、例えば重合工程で得られた重合体を塩基性条件下で加水分解することにより、脱炭酸を伴う標的分子の脱離を行えばよい。
【００４２】
ここで、共有結合型分子インプリンティングおよび非共有結合型分子インプリンティングの各々の利点を挙げる。共有結合型分子インプリンティングの利点は、標的分子と不飽和単量体（機能性モノマー）との複合体が比較的安定であること、結合部位の均一性が比較的高いこと、重合反応に用いる溶媒の選択において、溶媒による複合体の不安定化が起こりにくいために比較的制限が少ないこと等である。一方、非共有結合型分子インプリンティングの利点は、不飽和単量体（機能性モノマー）の重合前に、標的分子と不飽和単量体（機能性モノマー）との複合体を予め合成しておく必要がないこと、標的分子が比較的除去されやすいこと等である。
【００４３】
なお、以上の説明では、架橋剤との共重合を用いた分子インプリンティングによる製造方法について説明したが、分子認識ポリマーは、重縮合反応を利用した分子インプリンティングによっても製造可能である。
【００４４】
また、上記の説明では、分子認識ポリマーを、共有結合性分子認識ポリマーと非共有結合性分子認識ポリマーとの２種類に分類して説明したが、分子認識ポリマーは、標的分子に対して、可逆的な共有結合と可逆的な非共有結合との両方を形成するものであってもよい。このような結合モードの異なる分子認識の組み合わせは、選択性や親和性の増大が期待でき、応用範囲を広げることになる。このような分子認識ポリマーの例としては、標的分子に対して可逆的な配位結合を形成しうる錯体部位と、標的分子の水酸基に対して可逆的な水素結合を形成しうるカルボキシル基とを併せ持つ分子認識ポリマーが挙げられる。この分子認識ポリマーは、例えば、上記錯体部位を有する不飽和単量体と、カルボキシル基および光感応性部位を有する不飽和単量体と、架橋剤との共重合により得ることができる。
【００４５】
（具体例１）アゾベンゼン部位を有する光感応性インプリントポリマー
アゾベンゼンは、光照射により、容易にシス−トランス異性体間を相互変換することが知られている。このアゾベンゼンの二つのベンゼン環に、それぞれ、重合可能な二重結合と標的分子認識のための官能基を導入し、これを機能性モノマーとしてインプリントポリマーを合成すれば、光感応性インプリントポリマー光がえられる（実施例１・２参照）。この光感応性インプリントポリマーは、光照射によるアゾベンゼン部位の異性化にともない結合部位での分子認識能の変化が見られることが期待される。それゆえ、この光感応性インプリントポリマーは、標的分子の光応答性固相抽出（分離）に利用できることが期待される。
【００４６】
（具体例２）ベンゾスピロピラン基を持つ光感応性インプリントポリマー
スピロピラン類は、アゾベンゼンと並び代表的なフォトクロミック化合物である。互いに直行している二つのヘテロ環のうちの一つが紫外光の照射により開環し、その構造を大きく変える。従って、それぞれのヘテロ環を含む部分に、重合可能な二重結合と分子認識のための官能基を導入することにより、光照射による官能基の配向の変化により分子認識能が変化するインプリントポリマーを合成できると期待される。
【００４７】
ここでは、光異性化可能なベンゾスピロピラン基を導入した機能性モノマー（標的化合物と相互作用可能なモノマー）と、標的分子とを用いて、標的分子を吸脱着可能な光感応性インプリントポリマーを合成する。
【００４８】
まず、重合可能なビニル基やメタクリロイル基等の不飽和基と、標的分子と相互作用可能な官能基とを、ベンゾスピロピランに導入し、スピロピラン系機能性モノマーを合成する（図１（ａ））。例えば、メタクリロイルオキシメチル基を導入したサリチルアルデヒド誘導体を合成し、また、インドリン誘導体のクロル基を分子認識のための官能基（カルボキシル基やボロン酸基）に変換し、その後、これら二つの化合物をカップリングさせてスピロピラン系機能性モノマーを得る。
【００４９】
次いで、スピロピラン系機能性モノマーを標的分子と相互作用させて複合体を形成し、この状態で、架橋剤を加えて重合させる（図１（ｂ））。
【００５０】
最後に、標的分子を脱離させ、光感応性インプリントポリマーを得る（図１（ｃ））。
【００５１】
得られた光感応性インプリントポリマーには、標的分子と相補的な結合部位ができる。この結合部位は、光の入力でベンゾスピロピラン残基の異性化が誘起され、結合特性が変化する。そのため、この光感応性インプリントポリマーは、光のＯＮ−ＯＦＦあるいは波長の異なる光の照射により標的分子を吸着・脱着し、固相抽出が行えることが期待できる。したがって、上記光感応性インプリントポリマーには、固相抽出の際、溶媒を変えることなく、光のＯＮ−ＯＦＦあるいは波長の異なる光の照射で標的分子の濃縮および回収が繰り返し行え、分離科学の広い範囲に適用可能であると考えられる。
【００５２】
この分離手法は、特にトウモロコシ畑やゴルフ場などによく用いられるトリアジン除草剤の一つであるアトラジンを分離するのに有効であると考えられる。また、標的分子がアトラジンである場合、標的分子と相互作用可能な官能基としては、アトラジン１分子に対して複数の水素結合を形成して高い結合能を発現するカルボキシル基が好適である。アトラジンは、次式
【００５３】
【化２】

【００５４】
に示すように、カルボキシル基を持つ化合物と多点（４点）で水素結合を形成して、八員環キレート型の水素結合複合体を形成することが知られている。したがって、標的分子としてアトラジンを用い、カルボキシル基を持つ機能性モノマーを用いて分子インプリンティングを行えば、アトラジンを高選択的に吸着する光非共有結合性分子認識ポリマーが得られる。上記非共有結合性分子認識ポリマーを用いて固相抽出を行えば、アトラジンの回収を効率良く行うことができる。
【００５５】
上記のアトラジンと特異的に結合する非共有結合性分子認識ポリマーは、以下のようにして製造できる。すなわち、まず、図１（ａ）（Ｒ¹、Ｒ²はそれぞれ、水素原子または炭化水素基を示す）に示すように、メタクリロイルオキシメチル基およびカルボキシル基を有するベンゾスピロピラン系化合物（スピロピラン系機能性モノマー）と、付加重合可能な基を複数有する架橋剤（例えば、エチレングリコールジメタクリレート）とを用意し、これらをクロロホルム等の溶媒中でアトラジンと混合して、スピロピラン系機能性モノマーのカルボキシル基にアトラジンを結合させる。そして、これを２，２’−アゾビス（イソブチロニトリル）等の重合開始剤の存在下で重合した後、アトラジンを溶解しうる酸性溶液（例えばメタノール／酢酸）および溶媒（例えばメタノール）で洗浄することにより、アトラジンが除去され、非共有結合性分子認識ポリマーが得られる。
【００５６】
（具体例３）ペプチドを用いた光感応性分子認識ポリマー
水素結合等の非共有結合による分子認識を可能とする側鎖（標的分子に対して特異的にかつ可逆的に結合する官能基）と光異性化可能な基（光感応性部位）とを導入したペプチドを合成し、このペプチドをドーパミン（３，４−ジヒドロキシエチルアミン；カテコールアミン類の１種）等の標的分子の存在下で縮重合してオリゴペプチドとすることにより、光感応性分子認識ポリマーを得る。この光感応性分子認識ポリマーでは、標的分子と光感応性分子認識ポリマーとの相互作用の補強を図るために、チオール基等をさらに導入してもよい。
【００５７】
この光感応性分子認識ポリマーは、ドーパミン等の標的分子を認識する部位（官能基）近傍に光に応答する光感応性部位が導入されていれば、光により、認識部位の構造変化に伴う認識能の制御が可能となる。この光感応性分子認識ポリマーは、ドーパミン等の薬剤を担持して輸送し、光によるＯＮ−ＯＦＦにより薬剤を吸脱着可能であると考えられるため、ドラッグデリバリーシステムへの応用が期待できる（図６）。また、この光感応性分子認識ポリマーは、オリゴペプチドを主体としていることから、生分解性で生体適合性が高いと考えられる。また、この光感応性分子認識ポリマーは、回収、分解、再利用が可能であり、環境にやさしい材料である。
【００５８】
【実施例】
以下、本発明を実施例により詳細に説明する。なお、本発明は、以下の実施例の記載に限定されるものではなく、本発明の範囲内で種々の変更が可能である。
【００５９】
〔実施例１〕
本実施例では、メタクリロイルオキシ基と水酸基とを有するアゾベンゼンを合成し、これをコレステロールとの炭酸エステルとした機能性モノマーを調製し、この機能性モノマーをエチレングリコールジメタクリレートを架橋剤として重合することによりポリマーを合成し、得られたポリマー中の炭酸エステルを加水分解して、光感応性インプリントポリマーを合成した。この光感応性インプリントポリマーは、コレステロールと水素結合を形成しうるフェノール性水酸基と、光照射によりシス−トランス異性化するアゾベンゼン部位とを有することから、コレステロールを認識する分子認識能を持ち、かつ、光照射によりアゾベンゼン部位の異性化に伴って結合部位での分子認識能が変化することが期待される。
【００６０】
（Ｓ１）３，３’−ジヒドロキシアゾベンゼンの合成
まず、窒素雰囲気下、反応容器に、塩化銅二水和物（分子量170.48）１９．０５ｇ（０．１１ｍｏｌ）、３０重量％アンモニア水３３ｍｌおよび水２０ｍｌを投入した。次に、塩酸ヒドロキシルアミン（分子量69.49）２１．０７ｇ（０．３ｍｏｌ）を水６０ｍｌに溶解した溶液を滴下し、一価の銅錯体溶液を調製した。別の反応容器に、３−アミノフェノール（分子量109.13）１０．９３ｇ（０．１ｍｏｌ）と５ｍｏｌ／ｌ塩酸水溶液１００ｍｌとを投入し、氷冷した。ここに、亜硝酸ナトリウム（分子量69.00）７．０４ｇ（０．１ｍｏｌ）を水２０ｍｌに溶解した溶液を滴下し、ジアゾニウム塩溶液を生成させた。ここで生成したジアゾニウム塩溶液を水１３００ｍｌで希釈し、希釈したジアゾニウム塩溶液を氷冷した上記の銅錯体溶液に約１５分間かけて滴下し、そのまま３時間撹拌した。固体をろ別し、その固体を、３重量％アンモニア水５００ｍｌ、水２００ｍｌ、１ｍｏｌ／ｌ塩酸水溶液３００ｍｌ、および水２００ｍｌで洗浄した。洗浄した固体をカラムクロマトグラフィー（シリカゲル充填カラム、展開溶媒：酢酸エチル／ヘキサン＝２／３）で精製し、３，３’−ジヒドロキシアゾベンゼン（分子量214.22）４．９８ｇ（０．０２３ｍｏｌ）を得た。収率は４６％であった。
【００６１】
（Ｓ２）３’−ヒドロキシ−３−メタクリロイルオキシアゾベンゼンの調製
反応容器に、３，３’−ジヒドロキシアゾベンゼン（分子量214.22）１．２８ｇ（６ｍｍｏｌ）、塩化メチレン１５ｍｌ、およびピリジン３ｍｌを投入し、氷冷した。次いで、反応容器内の混合液を撹拌しながら、３０分かけて、塩化メタクリロイル（分子量104.54）０．３２ｇ（３ｍｍｏｌ）を塩化メチレン５ｍｌに溶解した溶液を滴下した。滴下終了後、０℃で３０分間、室温で３０分間撹拌した。溶媒を留去し、残渣を酢酸エチルに溶解して、水で洗浄した。酢酸エチル層を硫酸マグネシウムで乾燥させた後、溶媒を留去し、残渣をカラムクロマトグラフィー（シリカゲル充填カラム、展開溶媒：酢酸エチル／ヘキサン＝１／２）で精製し、３’−ヒドロキシ−３−メタクリロイルオキシアゾベンゼン（分子量270.29）０．５２ｇ（１．９ｍｍｏｌ）を得た。収率は６３％であった。
【００６２】
（Ｓ３）炭酸コレステリル３−（３−メタクリロイルオキシ）フェニルアゾフェニルの調製
反応容器に対し、３’−ヒドロキシ−３−メタクリロイルオキシアゾベンゼン０．２３ｇ（０．８ｍｍｏｌ）、テトラヒドロフラン１５ｍｌ、およびトリエチルアミン２ｍｌを投入し、氷冷した。次いで、反応容器内の混合液を撹拌しながら、この混合液に対し、氷冷によって液温を０〜５℃に保ちながらクロロギ酸コレステリル（分子量449.12）０．４０ｇ（０．９ｍｍｏｌ）を３０分間かけて滴下した。その後、液温を室温に戻し、室温で２０時間撹拌を行った。
【００６３】
反応混合液を分液ロートに移し、５重量％塩酸水溶液、水、５重量％水酸化ナトリウム水溶液、および水をそれぞれ加えて分液を行い、有機層を分離した。分離した有機層を硫酸マグネシウムで３０分間かけて乾燥させ、硫酸マグネシウムを濾過によって除去した。この濾過の濾液から溶媒を留去して得られた残渣をカラムクロマトグラフィー（シリカゲル充填カラム、展開溶媒：酢酸エチル／クロロホルム／ヘキサン＝１／２／１７）で精製し、炭酸コレステリル３−（３−メタクリロイルオキシ）フェニルアゾフェニル（分子量682.95）０．２１ｇ（０．３ｍｍｏｌ）を得た。収率は３８％であった。
【００６４】
（Ｓ４）ポリマーの調製
炭酸コレステリル３−（３−メタクリロイルオキシ）フェニルアゾフェニル６９．４ｍｇ（０．１ｍｍｏｌ）を、０．６４ｍｌのヘキサンおよび０．１６ｍｌのトルエン（溶媒）に溶解し、得られた溶液に、架橋剤としてのエチレングリコールジメタクリレート（分子量198.22）３７７ｍｇ（１．９ｍｍｏｌ）と、重合開始剤としての２，２’−アゾビス（イソブチロニトリル）（分子量164.21）５．０ｍｇ（０．０２ｍｍｏｌ）とを加えた。
【００６５】
そして、４５℃で１８時間かけて重合反応を行った。粗生成物を乳鉢で破砕し、メタノールで洗浄・乾燥した。これにより、ポリマー４１１ｍｇが得られた。
【００６６】
（Ｓ５）ポリマーの加水分解
得られたポリマーを、０．１ｍｍｏｌ／ｌの水酸化ナトリウムのメタノール溶液に懸濁させて、塩基性条件下で３時間かけて加水分解を行った。
【００６７】
これにより、脱炭酸を伴うコレステロールの脱離が起こり、結合部位にフェノール性水酸基が残った光感応性インプリントポリマー３７０ｍｇが得られた。
【００６８】
（Ｓ６）光感応性インプリントポリマーの分子認識能
クロロギ酸コレステリルに代えてクロロギ酸フェニルを用いる以外は、Ｓ２〜Ｓ４と同様の反応を行い、比較用のインプリントポリマーを合成した。
【００６９】
光感応性インプリントポリマーおよび比較用のインプリントポリマーをｎ−ヘキサン中に懸濁させて、コレステロールとの結合実験を行ったところ、光感応性インプリントポリマーは、比較用のインプリントポリマーに比べ、コレステロールの結合量が有意に多かった。このことから、光感応性インプリントポリマーが、コレステロールを認識する分子認識能を持っていることが確認された。
【００７０】
（Ｓ７）光感応性インプリントポリマーに結合させたコレステロールの紫外照射による脱離
この光感応性インプリントポリマーをｎ−ヘキサン中に懸濁させ、暗所でコレステロールを結合させた後、高圧水銀灯で波長３３０ｎｍの紫外線を１８分間照射したところ、結合していたコレステロールが遊離することが確認された。
【００７１】
〔実施例２〕
本実施例では、鋳型分子として４−ニトロフェニル−α−Ｄ−マンノピラノシドを用いて、結合定数の大きい環状ボロン酸エステル結合を持つ機能性モノマーを調製した。この機能性モノマーの合成スキームを以下に示す。
【００７２】
【化３】

【００７３】
そして、架橋剤として分解反応を起こさないジビニルベンゼンおよびスチレンを用いて上記機能性モノマーを重合することによりポリマーを合成し、得られたポリマー中のボロン酸エステルを加水分解して、光感応性インプリントポリマーを合成した。
【００７４】
この光感応性インプリントポリマーは、４−ニトロフェニル−α−Ｄ−マンノピラノシドと環状ボロン酸エステルを形成しうるボロン酸部位と、光照射によりシス−トランス異性化するアゾベンゼン部位とを有することから、４−ニトロフェニル−α−Ｄ−マンノピラノシドを認識する分子認識能を持ち、かつ、光照射によりアゾベンゼン部位の異性化に伴って結合部位での分子認識能が変化することが期待される。
（Ｓ１）アゾカップリング反応
２ｍｏｌ／ｌ塩酸水溶液８５ｍｌ中、０℃、１５分間の条件で、ｐ−ブロモアニリン（分子量172.02）４．３０ｇ（２５ｍｍｏｌ）を亜硝酸ナトリウム（分子量94.11）１．９０ｇ（２７．５ｍｍｏｌ）と反応させ、塩化ｐ−ブロモベンゼンジアゾニウムの水溶液を調製した。
【００７５】
続いて、得られた塩化ｐ−ブロモベンゼンジアゾニウム水溶液に対してフェノール（分子量94.11）２．３５ｇ（２５ｍｍｏｌ）および水酸化ナトリウム（分子量40.00）２ｇ（５０ｍｍｏｌ）を加え、０℃、５分間の条件でアゾカップリング反応を行った。
【００７６】
アゾカップリング反応生成物を粗生成物から単離することにより、４−ブロモ−４’−ヒドロキシアゾベンゼン（分子量277.125）２．７２ｇ（９．８ｍｍｏｌ）を得た。収率は３９％であった。
（Ｓ２）ボロン酸エステルに変換
反応容器に、４−ブロモ−４’−ヒドロキシアゾベンゼン０．５６ｇ（２．０ｍｍｏｌ）、［１，１'−ビス（ジフェニルホスフィノ）フェロセン］パラジウム（II）ジクロリド・塩化メチレン（１：１）錯体（分子量816.64）４４ｍｇ（０．０６ｍｍｏｌ）、ビス（ピナコラト）ジボロン（分子量253.94）０．５６ｇ（２．２ｍｍｏｌ）、酢酸カリウム（分子量98.15）０．３１ｇ（３．１ｍｍｏｌ）、および溶媒としての乾燥ＤＭＦ（Ｎ，Ｎ−ジメチルホルムアミド）１０ｍｌを投入し、反応混合物を窒素雰囲気下、８０℃で１８時間加熱した。反応の完了を確認した後、反応混合物をジエチルエーテルで希釈し、セライトで濾過し、水および食塩水で洗浄した。洗浄した溶液を硫酸マグネシウムで乾燥し、真空吸引により溶媒を除去した。その後、残渣を酢酸エチル／ヘキサン混合溶媒による再結晶で精製した。これにより、４’−ヒドロキシアゾベンゼン−４−ボロン酸ピナコラト（分子量324.19）０．６０ｇ（１．９ｍｍｏｌ）を得た。収率は９３％であった。
【００７７】
なお、ｄｐｐｆは配位子１，１’−ビス（ジフェニルホスフィノ）フェロセンを指す。
（Ｓ３）メタクリル酸エステル生成
４’−ヒドロキシアゾベンゼン−４−ボロン酸ピナコラト０．６１ｇ（１．９ｍｍｏｌ）を、塩化メチレン１０ｍｌに溶解させ、ピリジン１ｍｌを添加した。この溶液に対し、塩化メタクリロイル（分子量104.54）０．２７ｇ（２．６ｍｍｏｌ）を加えて、０℃で３０分間、室温で１時間撹拌することにより、エステル化反応を行った。
【００７８】
メタクリル酸エステルを粗生成物から単離することにより、４’−メタクリロイルオキシアゾベンゼン−４−ボロン酸ピナコラト（分子量392.26）０．５８ｇ（１．５ｍｍｏｌ）を得た。収率は７９％であった。
（Ｓ４）加水分解
４’−メタクリロイルオキシアゾベンゼン−４−ボロン酸ピナコラト０．５８ｇ（１．５ｍｍｏｌ）を塩酸／アセトン混合液中、室温、２４時間の条件で加水分解させた。
【００７９】
加水分解生成物を粗生成物から単離することにより、分子内にメタクリロイル基とボロン酸基とを有する機能性モノマーとしての４’−メタクリロイルオキシアゾベンゼン−４−ボロン酸（分子量310.12）０．１９ｇを得た。収率は４１％であった。
（Ｓ５）ボロン酸環状エステルモノマー調製
乾燥塩化メチレン２０ｍｌ中で、４’−メタクリロイルオキシアゾベンゼン−４−ボロン酸３１ｍｇ（０．１ｍｍｏｌ）と、鋳型分子としての４−ニトロフェニル−α−Ｄ−マンノピラノシド（分子量301.30）１５ｍｇ（０．０５ｍｍｏｌ）とを混合し、１８時間撹拌した（図２参照）。
【００８０】
溶媒を留去することにより、図２のＡに示す、４−ニトロフェニル−α−Ｄ−マンノピラノシドに対して２分子のボロン酸が環状エステルを形成したモノマー（ボロン酸環状エステルモノマー）（分子量849.41）４２ｍｇを得た。収率は１００％であった。このモノマーは、更なる精製は行わず、そのまま次のポリマー合成に使用した。
（Ｓ６）光感応性インプリントポリマーの調製
ボロン酸環状エステルモノマーを、図２に示すように、重合することで光感応性インプリントポリマーを調製した。
【００８１】
重合は、図３に示す操作により行った。すなわち、まず、ボロン酸環状エステルモノマー４２ｍｇ（０．０５ｍｍｏｌ）をテトラヒドロフラン０．８ｍｌに溶解し、得られた溶液に、架橋剤としてのジビニルベンゼン１９５ｍｇ（１．５ｍｍｏｌ）およびスチレン５２ｍｇ（０．５ｍｍｏｌ）と、重合開始剤としての２，２’−アゾビス（２,４−ジメチルバレロニトリル）５ｍｇ（０．０２ｍｍｏｌ）とを加えた。
【００８２】
そして、４５℃で１８時間かけて重合反応を行った。粗生成物を破砕し、メタノールで洗浄・乾燥することにより、ポリマーを単離した。これにより、ポリマー４１ｍｇが得られた。
（Ｓ７）環状ボロン酸エステルの加水分解
得られたポリマーを、図２および図４に示すように加水分解した。すなわち、得られたポリマーを、５重量％の塩酸を含むアセトン３０ｍｌに懸濁させることにより、酸性条件下で１６時間かけて環状ボロン酸エステルの加水分解を行った。
【００８３】
これにより、鋳型分子４−ニトロフェニル−α−Ｄ−マンノピラノシドがポリマーから脱離し、光感応性インプリントポリマーが得られた。
（Ｓ８）光感応性インプリントポリマーの紫外・可視吸収スペクトルの変化
光感応性インプリントポリマーに対し、波長３３１ｎｍの紫外線を１５分間照射したところ、図５に示すように、波長３３０ｎｍ付近の吸収（トランス体に起因する）が減少し、波長４４０ｎｍ付近の吸収（シス体に起因する）が増加した。これにより、トランス体が減少してシス体が増加すること、すなわちアゾベンゼン部位の異性化が起こっていることが確認された。
【００８４】
なお、図５に示す紫外・可視吸収スペクトルは、光感応性インプリントポリマーをクロロホルム中に懸濁させた状態で測定した。
【００８５】
【発明の効果】
本発明によれば、以上のように、標的分子と選択的に結合すると共に、光照射により物理・化学的変化を誘導できる光感応性を持ち、標的分子に対する吸脱着を光で制御可能な新規分子認識ポリマーを提供できる。
【００８６】
この分子認識ポリマーは、固相抽出の際、溶媒を変える煩雑な操作を行うことなく、光のＯＮ−ＯＦＦあるいは波長変更により標的分子の濃縮および回収が繰り返し行えるので、標的分子を固相抽出するための分離剤として極めて有用であると考えられる。また、固相抽出における溶媒の交換を不要としたことで、自動化が容易な固相抽出系を構築できる。
【００８７】
また、ドーパミン等の薬剤を標的分子とする分子認識ポリマーは、薬剤を担持して輸送し、光によるＯＮ−ＯＦＦにより薬剤を吸脱着可能であると考えられるため、ドラッグデリバリーシステムへの応用が期待できる。
【００８８】
また、本発明の製造方法によれば、上記の新規分子認識ポリマーを製造することができる。
【図面の簡単な説明】
【図１】本発明の一実施例における分子認識ポリマーの製造方法を示す図である。
【図２】本発明の他の実施例における分子認識ポリマーの製造方法を示す図である。
【図３】本発明の他の実施例における分子認識ポリマーの製造方法における、重合工程および脱離工程の操作・条件を示す図である。上記の図である。
【図４】本発明の他の実施例における分子認識ポリマーの製造方法の一工程を示す図である。
【図５】アゾベンゼン部位を持つ光感応性インプリントポリマーに対し、紫外線を照射したときの紫外・可視吸収スペクトルの変化を示す図である。
【図６】本発明の一形態における光感応性インプリントポリマーの分子認識能が外部刺激（光）により変化する様子を示す図である。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a molecular recognition polymer that selectively binds to a target molecule, and a method for producing a molecular recognition polymer using a molecular imprinting method.
[0002]
[Prior art]
As a technique for producing a molecular recognition polymer that specifically binds to a target molecule, an interesting technique called molecular imprinting is known in which the design of the functional site of the molecular recognition polymer is left to the molecule itself. The principle of molecular imprinting is very clear. The principle is that when a crosslinkable polymer having a specific binding site for the target molecule is synthesized, the target molecule is mixed into the polymer synthesis monomer and polymerized, and then added from the polymer formed after polymerization before polymerization. When the target molecule is washed out, a binding site complementary to the target molecule is formed in the polymer. If there is a characteristic functional group in the target molecule, a monomer (functional monomer) that interacts with the functional group by non-covalent bonding is polymerized together with a cross-linking agent that becomes a polymer matrix (hereinafter referred to as “non-covalent bonding type”). `` Molecular imprinting '') or by synthesizing a complex of a target molecule and a functional monomer in advance and polymerizing it with a crosslinking agent (hereinafter referred to as “covalent molecular imprinting”) This is a general technique. After polymerization, if the polymer swells and is washed under conditions where the target molecule is detached, it can interact with the target molecule in the state of the target molecule as a template after the target molecule in the polymer disappears A polymer in which the functional group remains and specifically binds to the target molecule (hereinafter referred to as “imprint polymer”) is obtained. That is, the functional group derived from the functional monomer is arranged in the matrix of the polymer derived from the crosslinking agent so as to recognize the characteristic functional group according to the shape of the target molecule. The area around this functionally localized functional group is a site that specifically binds to the target molecule.
[0003]
The inventors of the present invention have demonstrated that molecular recognition polymers that selectively adsorb various target molecules can be synthesized in a tailor-made manner by designing and synthesizing unique functional monomers in the molecular imprinting method. Patent References 1 to 5).
[0004]
In addition, the present inventors have shown that by adding some function to this molecular imprinting method, it is possible to create a new molecular recognition polymer that synchronizes with the binding of the target molecule and expresses the function. (See Non-Patent Documents 1 and 2).
[0005]
In addition, the inventors of the present application have applied the synthesized imprint polymer to separation techniques and sensors, and in particular, have demonstrated the usefulness by applying the imprint polymer to solid phase extraction (Non-Patent Documents 4 to 4). 6).
[0006]
The inventors of the present application have developed a robot that automatically performs polymer synthesis and evaluation of its binding ability, and has established the concept of combinatorial molecular imprinting (see Non-Patent Document 7).
[0007]
[Non-Patent Document 1]
Takeuchi, T., Mukawa, T., Matsui, J., Higashi, M., Shimizu, KD Molecularly Imprinted Polymers with Metalloporphyrin-Based Molecular Recognition Sites Coassembled with Methacrylic acid, Anal. Chem. 2001, 73, 3869-3874.
[0008]
[Non-Patent Document 2]
Matsui, J., Higashi, M., Takeuchi, T. Molecularly Imprinted Polymer as 9-Ethyladenine Receptor Having a Porphyrin-based Recognition Center, J. Am. Chem. Soc. 2000, 122, 5218-5219.
[0009]
[Non-Patent Document 3]
Takeuchi, T., Dobashi, A., Kimura, K. Molecular Imprinting of Biotin Derivatives and Its Application to Competitive Binding Assay Using Non-isotopic Labeled Ligands, Anal. Chem. 2000, 72, 2418-2422.
[0010]
[Non-Patent Document 4]
Molecularly Imprinted Polymers: Man made Mimics of Antibodies and Their Application in Analytical Chemistry, Sellergren, B. Ed., Elsevier, 2001.
[0011]
[Non-Patent Document 5]
Takeuchi, T., Haginaka, J. Separation and Sensing Based on Molecular Recognition Using Molecularly Imprinted Polymers, J. Chromatogr. B 1999, 728, 1-20.
[0012]
[Non-Patent Document 6]
Takeuchi, T., Matsui, J. Miniaturized Molecularly Imprinted Continuous Polymer Rods, J. High Resolution Chromatogr. 2000, 23, 44-46.
[0013]
[Non-Patent Document 7]
Takeuchi, T., Fukuma, D., Matsui, J. Combinatorial Molecular Imprinting: An Approach to Synthetic Polymer Receptors, Anal. Chem. 1999, 71, 285-290.
[0014]
[Problems to be solved by the invention]
However, in the solid-phase extraction of the target molecule by the molecular recognition polymer produced by the conventional molecular imprinting method, the target molecule is concentrated in a solvent condition that allows the target compound to be adsorbed on the adsorbent, and the target molecule is recovered in the target. Solvent conditions are used where molecules are likely to desorb from the adsorbent. This operation is cumbersome and hinders automation, and may cause degradation of the target molecule.
[0015]
If a molecule recognition material that can control adsorption / desorption to / from the target molecule by external stimulation can be provided, concentration and recovery of the target molecule can be performed by turning light on and off without performing complicated operations to change the solvent during solid phase extraction. It is thought that it can be repeated, and is considered extremely useful. However, there have been no examples of attempts to change the molecular recognition polymer in response to an external stimulus.
[0016]
The present invention has been made in view of the above-described conventional problems, and its purpose is to have a photosensitivity capable of selectively binding to a target molecule and inducing a physical / chemical change by light irradiation, and absorbing the target molecule. An object of the present invention is to provide a novel molecular recognition polymer whose desorption can be controlled by light and a method for producing the molecular recognition polymer using a molecular imprinting method.
[0017]
[Means for Solving the Problems]
In order to solve the above problems, the molecular recognition polymer of the present invention has a novel functional group that specifically and reversibly binds to a target molecule, and a photosensitive site that isomerizes by light irradiation. It is a molecular recognition polymer.
[0018]
The molecular recognition polymer can be selectively bonded to a target molecule by a functional group and can adsorb / desorb the target molecule. And since the said molecular recognition polymer has a photosensitivity part, isomerization (chemical change, physical change) of a photosensitivity part occurs by ON-OFF of light irradiation or irradiation of the light from which a wavelength differs, With this isomerization, the molecular structure in the vicinity of the functional group changes, and the binding ability (molecular recognition ability) of the functional group to the target molecule changes. Therefore, the molecular recognition polymer can adsorb and desorb the target molecule by controlling the molecular recognition ability of the functional group with respect to the target molecule by turning on / off light irradiation from the outside or changing the wavelength.
[0019]
DETAILED DESCRIPTION OF THE INVENTION
The molecular recognition polymer according to the present invention has a functional group that specifically and reversibly binds (interacts) to a target molecule (target compound), and a light-sensitive site that isomerizes by light irradiation.
[0020]
As the molecular recognition polymer, (1) a molecular recognition polymer having a covalent functional group that forms a reversible covalent bond specifically with respect to a target molecule and a photosensitive site that is isomerized by light irradiation (hereinafter referred to as a molecular recognition polymer). (Referred to as a “covalently binding molecule recognition polymer”), (2) a non-covalent functional group that forms a reversible non-covalent bond specifically to the target molecule, and photosensitivity that isomerizes by light irradiation. There is a molecular recognition polymer having a moiety (hereinafter, referred to as “non-covalent molecular recognition polymer”).
[0021]
In order for the target molecule to form a reversible bond with the covalently-bonded functional group of the covalently-bonded molecule recognition polymer, it is necessary that the covalent bond is easily generated and the target molecule can be detached. In view of this point, the above-described covalent bond must be not so strong, and the bond formation and dissociation must be easy and reversible. Examples of the reversible covalent bond include an ester bond between a boronic acid group and a cis-diol, an acetal bond between an aldehyde and a hydroxyl group, a ketal bond between a ketone and a hydroxyl group, and a Schiff base formed by a bond between an aldehyde and an amino group. And a coordination bond using a metal complex (for example, zinc porphyrin), an ester bond between a carboxyl group and a hydroxyl group, an amide bond between a carboxyl group and an amino group, and the like. Among these, reversible ester bond between a boronic acid group and a cis-diol is preferable because of its fast equilibrium.
[0022]
On the other hand, reversible non-covalent bonds that can be used for non-covalent molecular recognition polymers include hydrogen bond donating groups (hydrogen atoms on amide groups, hydrogen atoms on carboxyl groups, amino groups, hydroxyl groups, phenolic hydroxyl groups, etc. ) And hydrogen bond accepting groups (carbonyl group of amide group, carbonyl part of carboxyl group, nitrogen atom on nitrogen-containing heterocycle such as pyridine ring), ionic bond (cation and anion Electrostatic interaction). When the non-covalent molecular recognition polymer forms a hydrogen bond with the target molecule, it must form a multipoint hydrogen bond for highly selective and strong binding with the target molecule. It is preferable that a hydrogen bond is formed at 4 or more points. In addition, in order for the non-covalent molecular recognition polymer to bind to the target molecule in a highly selective and strong manner, the multipoint hydrogen bond may form an 8-membered chelate-type hydrogen bond complex. preferable. The combination of the compounds that form an 8-membered chelate-type hydrogen bond complex includes a compound having at least one amide group and / or carboxyl group and an amino group and a nitrogen atom adjacent to the nitrogen atom on the nitrogen-containing heterocyclic ring. And / or a combination with a compound having at least one carbonylamino group.
[0023]
The combination of the target molecule and the molecular recognition polymer is not particularly limited as long as it can form a reversible bond between them. That is, these combinations are between functional groups (such as boronic acid groups, hydroxyl groups, carboxyl groups, amino groups, phenolic hydroxyl groups, amide groups) possessed by molecular recognition polymers and functional groups (such as hydroxyl groups) possessed by target molecules. Any combination that forms a reversible bond may be used. Examples of combinations of target molecules and molecular recognition polymers include: (1) a compound having a catechol group having a cisdiol as a target molecule capable of forming a reversible ester bond, a polyhydric alcohol such as a sugar, and phenylboric acid (2) A combination of a compound that serves as a hydrogen donor and a compound that serves as a hydrogen acceptor (for example, atrazine as a target molecule and a carboxyl group that forms a hydrogen bond with atrazine And (3) a combination of a target molecule having a charge and a molecule recognition polymer having a charge opposite to that of the target molecule.
[0024]
In addition, the light-sensitive site may be any one that reversibly undergoes photoisomerization, that is, some chemical or physical change by light irradiation, but has a structure (conformation) by reversible isomerization by light irradiation. ) Having a large change, for example, an azobenzene moiety (azobenzene skeleton), a spiropyran moiety (spiropyran skeleton), and the like are suitable.
[Method for producing covalently binding molecule recognition polymer]
The covalent molecule recognition polymer can be produced by so-called covalent molecular imprinting. That is, the covalent molecule recognition polymer includes a preparation step of preparing an unsaturated monomer (also referred to as a functional monomer) having a covalent functional group and a photosensitive moiety, and the unsaturated monomer and target. A coupling step for binding the molecule by a reversible covalent bond, a polymerization step for copolymerizing an unsaturated monomer bound to the target molecule with a crosslinking agent to obtain a polymer, and a target molecule from the obtained polymer. And a desorption step for desorption.
[0025]
The unsaturated monomer is a functional group capable of forming a reversible covalent bond (the above-mentioned coordination bond, acetal-ketal bond, ester bond between boric acid and cis-diol, etc.) with the functional group of the target molecule. And a compound having an addition-polymerizable unsaturated bond and a light-sensitive site. Examples of the functional group capable of forming a reversible covalent bond with the functional group of the target molecule include a metal complex site capable of forming a coordinate bond with the oxygen atom of the target molecule, and a cyclic structure with the cis-diol site of the target molecule. The diboronic acid part etc. which can form an ester bond are mentioned.
[0026]
In the above preparation process, for example, an addition-polymerizable unsaturated group such as a vinyl group or a methacryloyl group can be added to a compound having a light-sensitive site to enable addition polymerization, and a covalent bond can be reversibly formed with a target molecule. An unsaturated monomer may be prepared by attaching a covalent functional group (etc.). Typical examples of the compound having a light-sensitive moiety include azobenzene and benzospiropyran. In addition, as a compound having a light-sensitive site, the structure does not change so much by light irradiation, but the color changes, diarylethene compounds that can be applied to rewritable recording materials, fluorides, salicylidene aniline compounds, etc. There are reversible photoresponsive compounds. In addition, carbon surrounded by three phenyl groups by light irradiation becomes a cation, and triphenylmethane cyanide whose polarity is greatly increased is also a compound having a light-sensitive site.
[0027]
In the polymerization step, an unsaturated monomer and a crosslinking agent are dissolved in a solvent, and a copolymerization reaction is performed using a polymerization initiator. The solvent also serves as a pore former to make the polymer porous. The solvent is not particularly limited as long as it can dissolve the target molecule and the unsaturated monomer, but chloroform is preferable because it dissolves each reagent well. As the polymerization initiator, 2,2′-azobis (isobutyronitrile), 2,2′-azobis (2,4-dimethylvaleronitrile) or the like can be used.
[0028]
The crosslinking agent used for the above addition polymerization is a compound having a group copolymerizable with a polymerizable group of the unsaturated monomer, for example, a vinyl group or a (meth) acryloyl group. Examples of the crosslinking agent are shown in the following formulas (1) to (20).
[0029]
[Chemical 1]

[0030]
As the crosslinking agent, those having a plurality of groups capable of addition polymerization are preferred. Examples of the crosslinking agent having a plurality of groups capable of addition polymerization include divinyl such as p-divinylbenzene represented by formula (4), o-divinylbenzene represented by formula (18), and m-divinylbenzene represented by formula (19). Examples include benzene-based crosslinking agents and (meth) acrylic acid-based crosslinking agents. Examples of the (meth) acrylic acid-based crosslinking agent include compounds having a plurality of (meth) acryloyl groups, such as ethylene glycol dimethacrylate (EDMA) represented by the formula (1), trimethylolpropane trimethacrylate (TRIM represented by the formula (2)). ), Trimethylolpropane triacrylate represented by formula (3), tetraethylene glycol diacrylate represented by formula (6), triethylene glycol dimethacrylate represented by formula (11), pentaerythritol tetraacrylate represented by formula (12), formula Pentaerythritol triacrylate shown in (13), bisphenol A dimethacrylate shown in formula (14), N, N′-methylenediacrylamide shown in formula (15), N, N′-1,4 shown in formula (16) -Phenylenediacrylamide, shown in formula (17) 3,5-bis (acryloylamide) benzoic acid, N shown in Equation (20), O-Bisuakuriroiru -L- phenylalaninol the like. As a crosslinking agent, a compound having only one vinyl group (vinyl chloride represented by formula (5), styrene represented by formula (7), phenyl vinyl sulfoxide represented by formula (9), phenyl vinyl sulfone represented by formula (10), etc. Or a compound having only one polymerizable group such as a compound having only one (meth) acryloyl group (for example, methyl methacrylate shown in the formula (8)) may be used. As the crosslinking agent, it is preferable to use a crosslinking agent having the same group as the polymerizable group of the unsaturated monomer. The crosslinking agent is preferably used in a large excess with respect to the unsaturated monomer in order to avoid crosstalk between binding sites. Specifically, it is preferable to use 5 mol or more of the crosslinking agent with respect to 1 mol of the unsaturated monomer.
[0031]
In the desorption step, the target molecule is desorbed by decomposing a reversible bond between the functional group and the target molecule in the polymer obtained in the polymerization step. For example, when the reversible bond is an ester bond, the target molecule may be eliminated by acid hydrolysis or alkali hydrolysis of the ester bond.
[0032]
[Method for producing non-covalently binding molecule recognition polymer]
The non-covalent molecular recognition polymer can be produced by so-called non-covalent molecular imprinting. That is, the non-covalent molecular recognition polymer includes a preparation step of preparing the unsaturated monomer having the non-covalent functional group and the light-sensitive site, and the presence of the target molecule in the unsaturated monomer. It can be manufactured by a method comprising a polymerization step of copolymerizing with a crosslinking agent under the above to obtain a polymer, and a desorption step of desorbing a target molecule from the obtained polymer.
[0033]
The unsaturated monomer is a compound having a functional group capable of forming a reversible non-covalent bond (such as a hydrogen bond) with a functional group of a target molecule, a polymerizable unsaturated bond, and a photosensitive site. It is. The functional group that can form a reversible non-covalent bond with the functional group of the target molecule includes a carbonyl group that can form a hydrogen bond with the amino group of the target molecule, or the nitrogen atom of the nitrogen-containing heterocyclic ring that the target molecule has. And a hydroxyl group and an amino group that can form a hydrogen bond.
[0034]
In the above preparation process, for example, an addition-polymerizable unsaturated group such as a vinyl group or a methacryloyl group is attached to a compound having a photosensitive moiety to enable addition polymerization, and a non-covalent bond is formed reversibly with the target molecule. A non-covalent functional group (carboxyl group, amino group, phenolic hydroxyl group, etc.) can be added to prepare an unsaturated monomer capable of addition polymerization.
[0035]
The above polymerization process is basically the same as the method for producing a covalent molecule recognition polymer, but in the case of noncovalent molecular imprinting, a complex of a target molecule and an unsaturated monomer is stably present. Therefore, the unsaturated monomer is preferably used in excess relative to the target molecule. Specifically, it is preferable to use 2 mol or more of the target molecule with respect to 1 mol of the target molecule. When the non-covalent bond is a hydrogen bond, the solvent used is preferably a halogen solvent such as chloroform that does not interfere with the hydrogen bond.
[0036]
In the desorption step, the target molecule is desorbed by washing with a solution (for example, an alcohol solvent) that can selectively dissolve the target molecule. Thereby, a non-covalently binding molecule recognition polymer is obtained.
[0037]
In the non-covalent molecular imprinting described above, the heterogeneity of the binding site formed may be a problem. Usually, in order to form a complex of a target molecule and an unsaturated monomer (functional monomer), the unsaturated monomer (functional monomer) is excessively added to the target molecule and polymerized. Naturally, the unsaturated monomer (functional monomer) involved in the binding site is limited, and the excess of added unsaturated monomer (functional monomer) is specific binding. It does not become a site and becomes non-specific binding. That is, the excess exists independently of the specific binding site, or exists as a binding site with low completeness and specificity.
[0038]
Therefore, in the non-covalent molecular imprinting described above, as a method for solving this problem, the non-specific molecular imprinting of the imprinted polymer obtained by the non-covalent molecular imprinting is followed by a non-specific molecular imprinting. You may perform the process (henceforth a "post-imprinting process") which chemically blocks a binding site and blocks. For example, for imprint polymers obtained by noncovalent molecular imprinting using unsaturated carboxylic acids as unsaturated monomers (functional monomers), coexistence of target molecules to protect specific binding sites Under the imprinted polymer, an unsaturated carboxylic acid-derived carboxylic acid residue that is not involved in the binding site may be methyl esterified with methyl iodide. This post-imprinting process can improve the selectivity of the non-covalently binding molecule-recognizing polymer with respect to the target molecule and improve the performance relatively easily.
[0039]
In the case where the target molecule has a hydroxyl group and the non-covalent molecule recognition polymer has a hydroxyl group that specifically forms a reversible hydrogen bond with the hydroxyl group of the target molecule, It can be produced by molecular imprinting. That is, in this case, the noncovalent molecular recognition polymer includes a preparation step of preparing a carbonate ester of the unsaturated monomer having the photosensitive moiety and the hydroxyl group and the target molecule having the hydroxyl group, and the carbonate ester. Is polymerized with a crosslinking agent to obtain a polymer, and the resulting polymer is desorbed by decarboxylation to desorb target molecules and liberate unsaturated monomer-derived hydroxyl groups. It can be manufactured by the method.
[0040]
In the preparation step, for example, the chloroformate ester of the target molecule may be reacted with an unsaturated monomer having a photosensitive site and a hydroxyl group in the presence of triethylamine.
[0041]
The polymerization step is the same as the method for producing the covalently bonded molecule recognition polymer. In the desorption step, for example, the target molecule accompanying decarboxylation may be desorbed by hydrolyzing the polymer obtained in the polymerization step under basic conditions.
[0042]
Here, advantages of each of covalent molecular imprinting and non-covalent molecular imprinting are listed. The advantages of covalent bond type molecular imprinting are that the complex of the target molecule and unsaturated monomer (functional monomer) is relatively stable, the uniformity of the binding site is relatively high, and it is used for the polymerization reaction. In the selection of the solvent, the complex is hardly destabilized by the solvent, so that there are relatively few restrictions. On the other hand, the advantage of noncovalent molecular imprinting is that a complex of target molecule and unsaturated monomer (functional monomer) is synthesized in advance before polymerization of unsaturated monomer (functional monomer). For example, the target molecule is relatively easy to remove.
[0043]
In the above description, the production method by molecular imprinting using copolymerization with a crosslinking agent has been described. However, the molecular recognition polymer can also be produced by molecular imprinting using a polycondensation reaction.
[0044]
In the above description, the molecular recognition polymer is classified into two types, that is, a covalent molecular recognition polymer and a non-covalent molecular recognition polymer. However, the molecular recognition polymer is reversible with respect to the target molecule. It may form both a covalent bond and a reversible non-covalent bond. Such a combination of molecular recognitions with different binding modes can be expected to increase selectivity and affinity, and the application range will be expanded. Examples of such molecular recognition polymers include a complex site capable of forming a reversible coordination bond with a target molecule and a carboxyl group capable of forming a reversible hydrogen bond with the hydroxyl group of the target molecule. Examples include molecular recognition polymers. This molecular recognition polymer can be obtained, for example, by copolymerization of an unsaturated monomer having the above complex site, an unsaturated monomer having a carboxyl group and a light sensitive site, and a crosslinking agent.
[0045]
(Example 1) Photosensitive imprint polymer having an azobenzene moiety
Azobenzene is known to easily interconvert between cis-trans isomers by light irradiation. If a double bond capable of polymerization and a functional group for target molecule recognition are introduced into the two benzene rings of azobenzene, respectively, and an imprint polymer is synthesized using this as a functional monomer, a photo-sensitive imprint polymer Light is obtained (see Examples 1 and 2). This photosensitive imprint polymer is expected to show a change in molecular recognition ability at the binding site as the azobenzene moiety is isomerized by light irradiation. Therefore, it is expected that this photosensitive imprint polymer can be used for photoresponsive solid phase extraction (separation) of target molecules.
[0046]
(Specific example 2) Photosensitive imprint polymer having benzospiropyran group
Spiropyrans are representative photochromic compounds along with azobenzene. One of the two heterocycles perpendicular to each other is opened by irradiation with ultraviolet light, and its structure is greatly changed. Therefore, by introducing a polymerizable double bond and a functional group for molecular recognition into each heterocycle-containing moiety, an imprinted polymer whose molecular recognition ability is changed by changing the orientation of the functional group by light irradiation. Is expected to be synthesized.
[0047]
Here, a photosensitive imprint polymer capable of adsorbing and desorbing a target molecule using a functional monomer (a monomer capable of interacting with the target compound) into which a photoisomerizable benzospiropyran group is introduced and a target molecule is prepared. Synthesize.
[0048]
First, an unsaturated group such as a polymerizable vinyl group or methacryloyl group and a functional group capable of interacting with a target molecule are introduced into benzospiropyran to synthesize a spiropyran-based functional monomer (FIG. 1 (a)). . For example, a salicylaldehyde derivative introduced with a methacryloyloxymethyl group is synthesized, and the chloro group of the indoline derivative is converted to a functional group for molecular recognition (carboxyl group or boronic acid group). Coupling yields a spiropyran functional monomer.
[0049]
Next, a spiropyran functional monomer interacts with the target molecule to form a complex, and in this state, a crosslinking agent is added to polymerize (FIG. 1B).
[0050]
Finally, the target molecule is desorbed to obtain a light-sensitive imprint polymer (FIG. 1 (c)).
[0051]
The resulting photosensitive imprint polymer has a binding site complementary to the target molecule. In this binding site, isomerization of the benzospiropyran residue is induced by the input of light, and the binding characteristics change. Therefore, this photosensitive imprint polymer can be expected to be able to perform solid-phase extraction by adsorbing and desorbing target molecules by light ON-OFF or light irradiation with different wavelengths. Therefore, the above-mentioned photosensitive imprint polymer can repeatedly concentrate and recover target molecules by light ON-OFF or light irradiation with different wavelengths without changing the solvent during solid phase extraction. It is considered applicable to a wide range.
[0052]
This separation method is considered to be particularly effective for separating atrazine, which is one of the triazine herbicides often used in corn fields and golf courses. When the target molecule is atrazine, the functional group capable of interacting with the target molecule is preferably a carboxyl group that forms a plurality of hydrogen bonds with one atrazine molecule and expresses high binding ability. Atrazine has the following formula
[0053]
[Chemical 2]

[0054]
As shown in the figure, it is known that a hydrogen bond is formed at multiple points (four points) with a compound having a carboxyl group to form an eight-membered chelate-type hydrogen bond complex. Therefore, when atrazine is used as a target molecule and molecular imprinting is performed using a functional monomer having a carboxyl group, a photo-noncovalent molecular recognition polymer that adsorbs atrazine with high selectivity can be obtained. If solid phase extraction is performed using the non-covalent molecular recognition polymer, atrazine can be efficiently recovered.
[0055]
The non-covalent molecular recognition polymer that specifically binds to the above-mentioned atrazine can be produced as follows. That is, first, FIG. ¹ , R ² Each represents a hydrogen atom or a hydrocarbon group), a benzospiropyran-based compound (spiropyran-based functional monomer) having a methacryloyloxymethyl group and a carboxyl group, and a crosslinking agent having a plurality of addition-polymerizable groups ( For example, ethylene glycol dimethacrylate) is prepared, and these are mixed with atrazine in a solvent such as chloroform, and atrazine is bonded to the carboxyl group of the spiropyran functional monomer. This is polymerized in the presence of a polymerization initiator such as 2,2′-azobis (isobutyronitrile), and then washed with an acidic solution (for example, methanol / acetic acid) that can dissolve atrazine and a solvent (for example, methanol). By doing so, atrazine is removed, and a non-covalently binding molecule recognition polymer is obtained.
[0056]
(Specific Example 3) Photosensitive molecular recognition polymer using peptide
Introduce side chains (functional groups that bind specifically and reversibly to target molecules) and photoisomerizable groups (photosensitive sites) that enable molecular recognition by non-covalent bonds such as hydrogen bonds By synthesizing this peptide in the presence of a target molecule such as dopamine (3,4-dihydroxyethylamine; one of catecholamines) to form an oligopeptide, a photosensitive molecule recognition polymer is obtained. obtain. In this photosensitive molecule recognition polymer, a thiol group or the like may be further introduced in order to reinforce the interaction between the target molecule and the photosensitive molecule recognition polymer.
[0057]
In this photosensitive molecule recognition polymer, if a light-sensitive site that responds to light is introduced in the vicinity of a site (functional group) that recognizes a target molecule such as dopamine, the light is recognized by the structural change of the recognition site. Can be controlled. This photosensitive molecule-recognizing polymer supports and transports a drug such as dopamine, and can be adsorbed and desorbed by light ON-OFF, so that it can be applied to a drug delivery system (FIG. 6). ). Moreover, since this photosensitive molecule recognition polymer is mainly composed of oligopeptides, it is considered to be biodegradable and highly biocompatible. The photosensitive molecule recognition polymer can be recovered, decomposed and reused, and is an environmentally friendly material.
[0058]
【Example】
Hereinafter, the present invention will be described in detail with reference to examples. In addition, this invention is not limited to description of a following example, A various change is possible within the scope of the present invention.
[0059]
[Example 1]
In this example, azobenzene having a methacryloyloxy group and a hydroxyl group is synthesized, a functional monomer is prepared by using this as a carbonate ester with cholesterol, and this functional monomer is polymerized using ethylene glycol dimethacrylate as a crosslinking agent. The polymer was synthesized by the method, and the carbonate ester in the obtained polymer was hydrolyzed to synthesize a photosensitive imprint polymer. This photosensitive imprint polymer has a phenolic hydroxyl group capable of forming a hydrogen bond with cholesterol and an azobenzene moiety that undergoes cis-trans isomerization by light irradiation, and thus has a molecular recognition ability to recognize cholesterol, and It is expected that the molecular recognition ability at the binding site changes with the isomerization of the azobenzene site by light irradiation.
[0060]
(S1) Synthesis of 3,3′-dihydroxyazobenzene
First, 19.05 g (0.11 mol) of copper chloride dihydrate (molecular weight 170.48), 33 ml of 30 wt% aqueous ammonia and 20 ml of water were charged into a reaction vessel under a nitrogen atmosphere. Next, a solution obtained by dissolving 21.07 g (0.3 mol) of hydroxylamine hydrochloride (molecular weight 69.49) in 60 ml of water was added dropwise to prepare a monovalent copper complex solution. In a separate reaction vessel, 10.93 g (0.1 mol) of 3-aminophenol (molecular weight 109.13) and 100 ml of 5 mol / l hydrochloric acid aqueous solution were charged and ice-cooled. A solution prepared by dissolving 7.04 g (0.1 mol) of sodium nitrite (molecular weight 69.00) in 20 ml of water was dropped to form a diazonium salt solution. The diazonium salt solution produced here was diluted with 1300 ml of water, and the diluted diazonium salt solution was added dropwise over about 15 minutes to the ice-cooled copper complex solution, followed by stirring for 3 hours. The solid was filtered off and the solid was washed with 500 ml of 3 wt% aqueous ammonia, 200 ml of water, 300 ml of 1 mol / l hydrochloric acid aqueous solution, and 200 ml of water. The washed solid was purified by column chromatography (silica gel packed column, developing solvent: ethyl acetate / hexane = 2/3) to obtain 4.98 g (0.023 mol) of 3,3′-dihydroxyazobenzene (molecular weight 214.22). . The yield was 46%.
[0061]
(S2) Preparation of 3'-hydroxy-3-methacryloyloxyazobenzene
A reaction vessel was charged with 1.28 g (6 mmol) of 3,3′-dihydroxyazobenzene (molecular weight 214.22), 15 ml of methylene chloride, and 3 ml of pyridine, and ice-cooled. Next, a solution prepared by dissolving 0.32 g (3 mmol) of methacryloyl chloride (molecular weight 104.54) in 5 ml of methylene chloride was added dropwise over 30 minutes while stirring the mixed solution in the reaction vessel. After completion of dropping, the mixture was stirred at 0 ° C. for 30 minutes and at room temperature for 30 minutes. The solvent was distilled off and the residue was dissolved in ethyl acetate and washed with water. After drying the ethyl acetate layer with magnesium sulfate, the solvent was distilled off, and the residue was purified by column chromatography (silica gel packed column, developing solvent: ethyl acetate / hexane = 1/2), and 3'-hydroxy-3 -0.52 g (1.9 mmol) of methacryloyloxyazobenzene (molecular weight 270.29) was obtained. The yield was 63%.
[0062]
(S3) Preparation of cholesteryl carbonate 3- (3-methacryloyloxy) phenylazophenyl
To the reaction vessel, 0.23 g (0.8 mmol) of 3′-hydroxy-3-methacryloyloxyazobenzene, 15 ml of tetrahydrofuran, and 2 ml of triethylamine were charged and ice-cooled. Next, while stirring the mixed solution in the reaction vessel, 0.40 g (0.9 mmol) of cholesteryl chloroformate (molecular weight 449.12) was maintained for 30 minutes while keeping the liquid temperature at 0 to 5 ° C. by cooling with ice. It was dripped over. Thereafter, the liquid temperature was returned to room temperature, and the mixture was stirred at room temperature for 20 hours.
[0063]
The reaction mixture was transferred to a separatory funnel, 5 wt% hydrochloric acid aqueous solution, water, 5 wt% sodium hydroxide aqueous solution and water were added to separate the layers, and the organic layer was separated. The separated organic layer was dried over magnesium sulfate for 30 minutes and the magnesium sulfate was removed by filtration. The residue obtained by distilling off the solvent from the filtrate of this filtration was purified by column chromatography (silica gel packed column, developing solvent: ethyl acetate / chloroform / hexane = 1/2/17) to give cholesteryl carbonate 3- (3 -Methacryloyloxy) phenylazophenyl (molecular weight 682.95) 0.21 g (0.3 mmol) was obtained. The yield was 38%.
[0064]
(S4) Preparation of polymer
Cholesteryl carbonate 3- (3-methacryloyloxy) phenylazophenyl 69.4 mg (0.1 mmol) was dissolved in 0.64 ml of hexane and 0.16 ml of toluene (solvent), and the resulting solution was used as a crosslinking agent. 377 mg (1.9 mmol) of ethylene glycol dimethacrylate (molecular weight 198.22) and 5.0 mg (0.02 mmol) of 2,2′-azobis (isobutyronitrile) (molecular weight 164.21) as a polymerization initiator were added. .
[0065]
And polymerization reaction was performed over 18 hours at 45 degreeC. The crude product was crushed with a mortar, washed with methanol and dried. This gave 411 mg of polymer.
[0066]
(S5) Hydrolysis of polymer
The obtained polymer was suspended in a 0.1 mmol / l sodium hydroxide methanol solution and hydrolyzed under basic conditions for 3 hours.
[0067]
As a result, elimination of cholesterol accompanied by decarboxylation occurred, and 370 mg of a photosensitive imprint polymer in which a phenolic hydroxyl group remained at the binding site was obtained.
[0068]
(S6) Molecular recognition ability of photosensitive imprint polymer
A comparative imprint polymer was synthesized by performing the same reaction as S2 to S4 except that phenyl chloroformate was used instead of cholesteryl chloroformate.
[0069]
When the photosensitive imprint polymer and the comparative imprint polymer were suspended in n-hexane and the binding experiment with cholesterol was conducted, the photosensitive imprint polymer was compared with the comparative imprint polymer. The amount of cholesterol bound was significantly higher. From this, it was confirmed that the photosensitive imprint polymer has a molecular recognition ability to recognize cholesterol.
[0070]
(S7) Desorption by ultraviolet irradiation of cholesterol bound to a photosensitive imprint polymer
After suspending this photosensitive imprint polymer in n-hexane, binding cholesterol in the dark, and then irradiating it with UV light at a wavelength of 330 nm with a high-pressure mercury lamp for 18 minutes, the bound cholesterol is released. Was confirmed.
[0071]
[Example 2]
In this example, 4-nitrophenyl-α-D-mannopyranoside was used as a template molecule to prepare a functional monomer having a cyclic boronic ester bond having a large binding constant. A synthesis scheme of this functional monomer is shown below.
[0072]
[Chemical 3]

[0073]
Then, a polymer is synthesized by polymerizing the functional monomer using divinylbenzene and styrene that do not cause a decomposition reaction as a crosslinking agent, and the boronic acid ester in the obtained polymer is hydrolyzed to produce a photo-sensitive indium. A print polymer was synthesized.
[0074]
Since this photosensitive imprint polymer has a boronic acid moiety capable of forming a cyclic boronic acid ester with 4-nitrophenyl-α-D-mannopyranoside, and an azobenzene moiety that undergoes cis-trans isomerization by light irradiation, It has molecular recognition ability for recognizing 4-nitrophenyl-α-D-mannopyranoside, and it is expected that the molecular recognition ability at the binding site changes with isomerization of the azobenzene site by light irradiation.
(S1) Azo coupling reaction
In 85 ml of 2 mol / l hydrochloric acid aqueous solution, 4.30 g (25 mmol) of p-bromoaniline (molecular weight 172.02) was reacted with 1.90 g (27.5 mmol) of sodium nitrite (molecular weight 94.11) at 0 ° C. for 15 minutes. An aqueous solution of p-bromobenzenediazonium chloride was prepared.
[0075]
Subsequently, 2.35 g (25 mmol) of phenol (molecular weight 94.11) and 2 g (50 mmol) of sodium hydroxide (molecular weight 40.00) were added to the obtained aqueous p-bromobenzenediazonium chloride solution, and the conditions were 0 ° C. and 5 minutes. An azo coupling reaction was performed.
[0076]
The azo coupling reaction product was isolated from the crude product to obtain 2.72 g (9.8 mmol) of 4-bromo-4′-hydroxyazobenzene (molecular weight 277.125). The yield was 39%.
(S2) Conversion to boronic acid ester
In a reaction vessel, 4-bromo-4′-hydroxyazobenzene 0.56 g (2.0 mmol), [1,1′-bis (diphenylphosphino) ferrocene] palladium (II) dichloride / methylene chloride (1: 1) complex (Molecular weight 816.64) 44 mg (0.06 mmol), bis (pinacolato) diboron (molecular weight 253.94) 0.56 g (2.2 mmol), potassium acetate (molecular weight 98.15) 0.31 g (3.1 mmol), and dry DMF as solvent 10 ml of (N, N-dimethylformamide) was added and the reaction mixture was heated at 80 ° C. for 18 hours under a nitrogen atmosphere. After confirming the completion of the reaction, the reaction mixture was diluted with diethyl ether, filtered through celite and washed with water and brine. The washed solution was dried with magnesium sulfate, and the solvent was removed by vacuum suction. Thereafter, the residue was purified by recrystallization using an ethyl acetate / hexane mixed solvent. As a result, 0.60 g (1.9 mmol) of 4′-hydroxyazobenzene-4-boronic acid pinacolato (molecular weight: 324.19) was obtained. The yield was 93%.
[0077]
Dppf represents the ligand 1,1′-bis (diphenylphosphino) ferrocene.
(S3) Methacrylate formation
0.61 g (1.9 mmol) of 4′-hydroxyazobenzene-4-boronic acid pinacolato was dissolved in 10 ml of methylene chloride, and 1 ml of pyridine was added. To this solution, 0.27 g (2.6 mmol) of methacryloyl chloride (molecular weight 104.54) was added and stirred at 0 ° C. for 30 minutes and at room temperature for 1 hour to carry out an esterification reaction.
[0078]
By isolating the methacrylic acid ester from the crude product, 48-methacryloyloxyazobenzene-4-boronic acid pinacolato (molecular weight 392.26) 0.58 g (1.5 mmol) was obtained. The yield was 79%.
(S4) Hydrolysis
0.58 g (1.5 mmol) of 4′-methacryloyloxyazobenzene-4-boronic acid pinacolato was hydrolyzed in a hydrochloric acid / acetone mixture at room temperature for 24 hours.
[0079]
By isolating the hydrolysis product from the crude product, 0.19 g of 4′-methacryloyloxyazobenzene-4-boronic acid (molecular weight 310.12) as a functional monomer having a methacryloyl group and a boronic acid group in the molecule is obtained. Got. The yield was 41%.
(S5) Preparation of boronic acid cyclic ester monomer
In 20 ml of dry methylene chloride, 31 mg (0.1 mmol) of 4′-methacryloyloxyazobenzene-4-boronic acid and 15 mg (0.05 mmol) of 4-nitrophenyl-α-D-mannopyranoside (molecular weight 301.30) as a template molecule And stirred for 18 hours (see FIG. 2).
[0080]
By distilling off the solvent, a monomer (boronic acid cyclic ester monomer) in which two molecules of boronic acid formed a cyclic ester with respect to 4-nitrophenyl-α-D-mannopyranoside (molecular weight 849.41) shown in FIG. ) 42 mg was obtained. The yield was 100%. This monomer was used for the next polymer synthesis without further purification.
(S6) Preparation of light-sensitive imprint polymer
As shown in FIG. 2, a photosensitive imprint polymer was prepared by polymerizing a boronic acid cyclic ester monomer.
[0081]
The polymerization was performed by the operation shown in FIG. That is, first, 42 mg (0.05 mmol) of a boronic acid cyclic ester monomer was dissolved in 0.8 ml of tetrahydrofuran, and 195 mg (1.5 mmol) of divinylbenzene and 52 mg (0.5 mmol) of styrene as a crosslinking agent were added to the resulting solution. Then, 5 mg (0.02 mmol) of 2,2′-azobis (2,4-dimethylvaleronitrile) as a polymerization initiator was added.
[0082]
And polymerization reaction was performed over 18 hours at 45 degreeC. The crude product was crushed, and the polymer was isolated by washing with methanol and drying. This gave 41 mg of polymer.
(S7) Hydrolysis of cyclic boronic acid ester
The resulting polymer was hydrolyzed as shown in FIGS. That is, the cyclic boronic acid ester was hydrolyzed for 16 hours under acidic conditions by suspending the obtained polymer in 30 ml of acetone containing 5 wt% hydrochloric acid.
[0083]
As a result, the template molecule 4-nitrophenyl-α-D-mannopyranoside was detached from the polymer, and a light-sensitive imprinted polymer was obtained.
(S8) Change in UV-visible absorption spectrum of photosensitive imprint polymer
When the photosensitive imprint polymer was irradiated with ultraviolet light having a wavelength of 331 nm for 15 minutes, as shown in FIG. 5, the absorption near the wavelength of 330 nm (due to the trans form) decreased and the absorption near the wavelength of 440 nm (cis) Due to the body) increased. As a result, it was confirmed that the trans isomer decreased and the cis isomer increased, that is, the azobenzene moiety was isomerized.
[0084]
Note that the ultraviolet / visible absorption spectrum shown in FIG. 5 was measured in a state where the photosensitive imprint polymer was suspended in chloroform.
[0085]
【The invention's effect】
According to the present invention, as described above, a novel compound capable of selectively binding to a target molecule, having photosensitivity capable of inducing a physical / chemical change by light irradiation, and controlling adsorption / desorption with respect to the target molecule by light. Molecular recognition polymers can be provided.
[0086]
This molecular recognition polymer can be used for solid-phase extraction of the target molecule because it can be repeatedly concentrated and recovered by turning the light on and off or changing the wavelength without performing complicated operations to change the solvent during solid-phase extraction. Therefore, it is considered extremely useful as a separating agent. In addition, by eliminating the need for solvent exchange in solid phase extraction, a solid phase extraction system that can be easily automated can be constructed.
[0087]
In addition, molecular recognition polymers that use drugs such as dopamine as target molecules are thought to be capable of adsorbing and desorbing drugs by carrying and transporting the drug and turning it on and off by light, so application to drug delivery systems is expected. it can.
[0088]
Moreover, according to the manufacturing method of this invention, said novel molecule | numerator recognition polymer can be manufactured.
[Brief description of the drawings]
FIG. 1 is a diagram showing a method for producing a molecular recognition polymer in one embodiment of the present invention.
FIG. 2 is a diagram showing a method for producing a molecular recognition polymer in another embodiment of the present invention.
FIG. 3 is a diagram showing operations and conditions of a polymerization step and a desorption step in a method for producing a molecule recognition polymer in another example of the present invention. FIG.
FIG. 4 is a diagram showing one step of a method for producing a molecular recognition polymer according to another embodiment of the present invention.
FIG. 5 is a diagram showing a change in ultraviolet / visible absorption spectrum when a photosensitive imprint polymer having an azobenzene moiety is irradiated with ultraviolet rays.
FIG. 6 is a diagram showing a state in which the molecular recognition ability of a light-sensitive imprint polymer according to an embodiment of the present invention changes due to external stimulation (light).

Claims (9)

It has a functional group that specifically and reversibly binds to the target molecule, and a light-sensitive site that isomerizes by light irradiation ,
The functional group includes a covalent functional group that forms a specifically reversible covalent bond to a target molecule,
A preparation step of preparing an unsaturated monomer having the covalent bond functional group and the light-sensitive moiety;
A binding step of binding the unsaturated monomer and the target molecule by a reversible covalent bond;
A polymerization step of copolymerizing an unsaturated monomer bonded to a target molecule with a crosslinking agent to obtain a polymer;
An imprint polymer obtained by a method for producing an imprint polymer including a desorption step of desorbing a target molecule from the obtained polymer. The target molecule has a plurality of hydroxyl groups,
2. The imprint polymer according to claim 1, wherein the covalent bond functional group includes a boronic acid group that forms a cyclic boronic acid ester by bonding with a plurality of hydroxyl groups of the target molecule . It has a functional group that specifically and reversibly binds to the target molecule, and a light-sensitive site that isomerizes by light irradiation,
The functional group includes a non-covalent functional group that forms a specifically reversible non-covalent bond to a target molecule,
A preparation step of preparing an unsaturated monomer having the non-covalent functional group and the photo-sensitive moiety;
A polymerization step of copolymerizing the unsaturated monomer with a crosslinking agent in the presence of a target molecule to obtain a polymer;
An imprint polymer obtained by a method for producing an imprint polymer including a desorption step of desorbing a target molecule from the obtained polymer. It has a functional group that specifically and reversibly binds to the target molecule, and a light-sensitive site that isomerizes by light irradiation,
The functional group includes a non-covalent functional group that forms a specifically reversible non-covalent bond to a target molecule,
The target molecule has a hydroxyl group,
The non-covalent functional group includes a hydroxyl group that specifically forms a reversible hydrogen bond with the hydroxyl group of the target molecule,
A preparation step of preparing a carbonate ester of the unsaturated monomer having the photosensitive moiety and the hydroxyl group and the target molecule having the hydroxyl group;
A polymerization step of copolymerizing the carbonate ester with a crosslinking agent to obtain a polymer;
An imprint polymer obtained by a method for producing an imprint polymer comprising a desorption step of desorbing a target molecule by decarboxylation and liberating a hydroxyl group derived from an unsaturated monomer from the obtained polymer. Print polymer. The imprinted polymer according to any one of claims 1 to 4, wherein the light-sensitive moiety includes an azobenzene moiety . The imprinted polymer according to any one of claims 1 to 4, wherein the light-sensitive portion includes a spiropyran portion . A method for producing an imprinted polymer according to claim 1,
A preparation step of preparing an unsaturated monomer having the covalent bond functional group and the light-sensitive moiety;
A binding step of binding the unsaturated monomer and the target molecule by a reversible covalent bond;
A polymerization step of copolymerizing an unsaturated monomer bonded to a target molecule with a crosslinking agent to obtain a polymer;
A method for producing an imprint polymer, comprising a desorption step of desorbing a target molecule from the obtained polymer. A method for producing an imprinted polymer according to claim 3 ,
A preparation step of preparing an unsaturated monomer having the non -covalent functional group and the photo-sensitive moiety;
A polymerization step of copolymerizing the unsaturated monomer with a crosslinking agent in the presence of a target molecule to obtain a polymer;
A method for producing an imprint polymer, comprising a desorption step of desorbing a target molecule from the obtained polymer. A method for producing an imprinted polymer according to claim 4 ,
A preparation step of preparing a carbonate ester of the unsaturated monomer having the photosensitive moiety and the hydroxyl group and the target molecule having the hydroxyl group;
A polymerization step of copolymerizing the carbonate ester with a crosslinking agent to obtain a polymer;
A method for producing an imprint polymer, comprising a desorption step of desorbing a target molecule from the obtained polymer by decarboxylation to liberate a hydroxyl group derived from an unsaturated monomer .

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