JP2004284991A - N-acylamine compound and medical composition using the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a biodegradable surface-covered compound which delivers drugs etc. to tissues or cells aimed at in the body; and a medical carrier using the same. <P>SOLUTION: The N-acylamine compound is represented by general formula (1) [wherein R<SB>1</SB>is a 1-23C alkyl or an alkenyl having at least one double bond; R<SB>2</SB>is H or a 1-18C alkyl or alkenyl; R<SB>3</SB>is a 1-18C alkylene; A is O, S, COO, OCO, CONR<SB>4</SB>, NR<SB>4</SB>CO, NHCOO, OCONH, or NHCONH (wherein R<SB>4</SB>is H or a 1-18C alkyl or alkenyl); and X is an organism-derived substance]. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

【００２９】
（ここでＲ_１は炭素数１〜２３のアルキル基または少なくとも一つの二重結合を有するアルケニル基、Ｒ_２は水素または炭素数１〜１８のアルキル基またはアルケニル基、Ｒ３は炭素数１〜１８のアルキレン基、ＡはＯ、Ｓ、ＣＯＯ、ＯＣＯ、ＣＯＳ、ＳＣＯ、ＣＯＮＲ４、ＮＲ４ＣＯ、ＮＨＣＯＯ、ＯＣＯＮＨ、ＮＨＣＯＳ、ＳＣＯＮＨ、ＮＨＣＯＮＨのいずれか（ここでＲ４は、水素または炭素数１〜１８のアルキル基またはアルケニル基）、Ｘは生体由来物質）で表されるＮ−アシルアミン化合物。
（２）一般式（１）中のＲ_２が水素であることを特徴とする（１）に記載のＮ−アシルアミン化合物。
（３）一般式（１）中のＲ_１のアルキル基またはアルケニル基が炭素数１２〜２３の直鎖状であることを特徴とする（１）または（２）に記載のＮ−アシルアミン化合物。
（４）一般式（１）中のＲ_１がアルケニル基であり、かつ二重結合の数が１〜５であることを特徴とする（１）〜（３）のいずれかに記載のＮ−アシルアミン化合物。
（５）一般式（１）中のＲ３が炭素数１〜４のアルキレン基であることを特徴とする（４）に記載のＮ−アシルアミン化合物。
（６）一般式（１）中のＸが糖から選ばれることを特徴とする（１）〜（５）のいずれかに記載のＮ−アシルアミン化合物。
（７）一般式（１）中のＸがグルコース、マンノース、マンノース−６−リン酸、フコース、ガラクトース、グルコサミン、ラクトサミン、ガラクトサミン、Ｎ−アセチルグルコサミン、Ｎ−アセチルガラクトサミン、グルコン酸、グルクロン酸、ガラクツロン酸、ラクトース、マルトース、イソマルトース、スクロース、セロビオース、トレハロース、ラクトビオン酸、アポＢ、トランスフェリン、タフツシン、免疫グロブリン、葉酸であることを特徴とする（１）〜（６）のいずれかに記載のＮ−アシルアミン化合物。
（８）生分解性であることを特徴とする（１）〜（７）のいずれかに記載のＮ−アシルアミン化合物。
（９）（１）〜（８）のいずれかに記載の少なくとも一つのＮ−アシルアミン化合物を、少なくとも表面に有する粒子。
（１０）該粒子がリポソームであることを特徴とする（９）に記載の粒子。
（１１）（１）〜（８）のいずれかに記載の少なくとも一つのＮ−アシルアミン化合物を基材粒子の表面にコーティングしてなることを特徴とする請求項９に記載の粒子。
（１２）該基材粒子が生分解性材料からなることを特徴とする（９）〜（１１）のいずれかに記載の粒子。
（１３）該粒子のサイズが０．１ｎｍ〜５ｍｍの範囲にあることを特徴とする（９）〜（１２）のいずれかに記載の粒子。
（１４）該粒子中に少なくとも生理活性物質、薬剤、遺伝子のいずれか１つを含有してなることを特徴とする（９）〜（１３）のいずれかに記載の粒子。
（１５）該生理活性物質が、増殖因子、サイトカイン、ホルモン、血管作動性物質から選ばれる少なくとも１種であることを特徴とする（１４）に記載の粒子。
（１６）該薬剤がステロイド、造影剤、抗癌薬、抗炎症薬、抗アレルギー薬、循環器薬、降圧薬、昇圧薬、抗血小板薬、抗凝固薬、向精神薬、疼痛治療薬、抗生物質、抗ウイルス薬、消化器薬、泌尿器薬、内分泌薬、蛍光試薬から選ばれる少なくとも１種であることを特徴とする（１４）に記載の粒子。
（１７）該遺伝子がＤＮＡ、ＲＮＡから選ばれる少なくとも１種であることを特徴とする（１４）に記載の粒子。
（１８）（１）〜（８）のいずれかに記載のＮ−アシルアミン化合物と生理活性物質または薬剤を水溶性有機溶媒に溶解した後、水中に滴下し、有機溶媒を除去し、遠心分離または濾過のいずれかで被覆粒子を分離することを特徴とする粒子の製造方法。
（１９）（１）〜（８）のいずれかに記載のＮ−アシルアミン化合物と生理活性物質または薬剤を非水溶性有機溶媒に溶解し、水中に滴下した後、有機溶媒を除去し、遠心分離または濾過のいずれかで被覆粒子を分離することを特徴とする粒子の製造方法。
（２０）粒子の原料物質と生理活性物質または薬剤を非水溶性有機溶媒に溶解した後、（１）〜（８）のいずれかに記載のＮ−アシルアミン化合物を溶解した水溶液中に滴下し、有機溶媒を除去し、遠心分離または濾過のいずれかで被覆粒子を分離することを特徴とする被覆粒子の製造方法。
（２１）（１）〜（８）のいずれかに記載のＮ−アシルアミン化合物と粒子の原料物質と生理活性物質または薬剤を非水溶性有機溶媒に溶解した後、水中に滴下し、有機溶媒を除去し、遠心分離または濾過のいずれかで被覆粒子を分離することを特徴とする被覆粒子の製造方法。
（２２）超音波処理を行いながら水中に滴下することを特徴とする請求項（１８）〜（２２）に記載の粒子の製造方法。
（２３）攪拌を行いながら水中に滴下することを特徴とする請求項（１８）〜（２３）に記載の粒子の製造方法。
（２４）減圧下で有機溶媒を除去することを特徴とする請求項（１８）〜（２４）に記載の粒子の製造方法。
（２５）（１）〜（８）のいずれかに記載のＮ−アシルアミン化合物と生理活性物質または薬剤を水溶性有機溶媒に溶解した後、透析チューブに充填し、水に対して透析を行って有機溶媒を除去し、遠心分離または濾過のいずれかで被覆粒子を分離することを特徴とする被覆粒子、リポソーム、ミセルのいずれかの製造方法。
（２６）（１）〜（８）のいずれかに記載のＮ−アシルアミン化合物を少なくとも表面有する医療用器材。
（２７）該医療用組成物がカテーテルであることを特徴とする（２６）に記載の医療用器材。
（２８）該医療用組成物が細胞治療システム器材であることを特徴とする（２６）に記載の医療用器材。
（２９）該医療用組成物が検査診断薬用器材であることを特徴とする（２６）に記載の医療用器材。
（３０）該医療用組成物がインプラントデバイスであることを特徴とする（２６）に記載の医療用器材。
（３１）該医療用組成物が体外循環システムであることを特徴とする（２６）に記載の医療用器材。」
【００３０】
【発明の実施の形態】
以下、本発明を詳細に説明する。
【００３１】
本発明のＮ−アシルアミン化合物は、下記一般式
【００３２】
【化３】

【００３３】
（ここでＲ_１は炭素数１〜２３のアルキル基または少なくとも一つの二重結合を有するアルケニル基、Ｒ_２は水素または炭素数１〜１８のアルキル基またはアルケニル基、Ｒ_３は炭素数１〜１８のアルキレン基、ＡはＯ、Ｓ、ＣＯＯ、ＯＣＯ、ＣＯＳ、ＳＣＯ、ＣＯＮＲ４、ＮＲ４ＣＯ、ＮＨＣＯＯ、ＯＣＯＮＨ、ＮＨＣＯＳ、ＳＣＯＮＨ、ＮＨＣＯＮＨのいずれか（ここでＲ４は、水素または炭素数１〜１８のアルキル基またはアルケニル基）、Ｘは生体由来物質）で表される。
【００３４】
本発明のＮ−アシルアミン化合物は生体由来の物質からなることを特徴としている。従って、このＮ−アシルアミン化合物の構成要素の構造に関しては、基本的には生体物質由来であればよく、特に限定されるものではない。一般式（１）中のＲ_１はこの分子の疎水性部分を形成し、炭素数１〜２４のアルキル基またはアルケニル基が好ましく、枝分かれしていてもよい。生体内に高い割合で存在するという理由で、炭素数１１〜１９が最も好ましく、更に直鎖状であることが好ましい。Ｒ_１がアルケニル基の場合には、生体内に高い割合で存在するという理由で、２重結合が１〜５個含まれていることが好ましい。Ｒ_２は水素または炭素数１〜１８のアルキル基またはアルケニル基であることが好ましい。Ｒ_１とＲ_３とを結合する−ＣＯＮＲ_２−結合はアミド結合であり、体温付近の温度では酵素が無いと加水分解反応が起こりにくい。この結合が酵素によって加水分解切断されるためには、Ｒ_２は水素であることが最も望ましい。また、Ｒ_２が水素の場合、アミド結合は互いに水素結合することが知られているので、この分子を基材表面にコーティングすると、基材表面で分子が相互作用し合い高い表面密度が得られる。Ｒ３は炭素数１〜１８の中から自由に選べるが、生体内に高い割合で存在するという理由で、炭素数２〜４のアルキレン基であることが好ましい。Ｘも生体由来物質から成ることを特徴とする物質で、被覆される表面を特徴づけるリガンドとなる。Ｘの具体例としては、アミノ酸、オリゴペプチド、ポリペプチド、抗体、抗体の一部、受容体、酵素、グルコース、マンノース、マンノース−６−リン酸、ガラクトース、グルコサミン、ラクトサミン、ガラクトサミン、Ｎ−アセチルグルコサミン、Ｎ−アセチルガラクトサミン、グルコン酸、グルクロン酸、ガラクツロン酸などの単糖、ラクトース、マルトース、イソマルトース、スクロース、セロビオース、トレハロース、シアリルルイスＸ、シアリルルイスａなどのオリゴ糖、ヘパリン、低分子量ヘパリン、ヒアルロン酸、デルマタン硫酸、コンドロイチン硫酸、ケラタン硫酸、シンデカンなどの多糖等が挙げられるが、これに限定されるものではない。
【００３５】
特に肝臓あるいは肝実質細胞との親和性を付与するためにこの化合物を用いる場合には、Ｘとしてガラクトース、ガラクトサミン、非還元末端にガラクトースまたはガラクトサミンを有するオリゴ糖または多糖、アポＢが好ましく用いられる。糖鎖は分岐していても良く、分岐鎖のそれぞれの非還元末端にガラクトースまたはガラクトサミンが存在していることが好ましい。
【００３６】
特にマクロファージとの親和性を付与するためにこの化合物を用いる場合には、Ｘとしてガラクトサミン、非還元末端にガラクトサミンを有するオリゴ糖または多糖、アポＢが好ましく用いられる。糖鎖は分岐していても良く、分岐鎖のそれぞれの非還元末端にガラクトサミンが存在していることが好ましい。また、タフツシンも好ましく用いられる。
【００３７】
特にＴ細胞との親和性を付与するためにこの化合物を用いる場合には、ＸとしてがＨＩＶウイルス由来ｇｐ１２０ｇが好ましく用いられる。
【００３８】
特に癌細胞との親和性を付与するためにこの化合物を用いる場合には、Ｘとしてトランスフェリン、ＥＧＦ、タフツシン、葉酸が好ましく用いられる。
【００３９】
一般式１中のＲ_１〜Ｒ_３とＸの構造を選ぶことによって、Ｎ−アシルアミン化合物の溶媒に対する溶解性も自由に変えることができる。
【００４０】
生体由来物質ＸとＲ_３を連結する結合は、Ｘの化学構造や配向性、分解性に応じて自由に選択することができ、生体内で分解されやすいという点で、加水分解を受けやすいエステル結合やウレイド結合、酵素分解を受けやすいペプチド結合に代表されるアミド結合、グルコシド結合、ガラクトシド結合に代表されるエーテル結合やチオエーテル結合が好ましく、すなわちＡの具体例としてはＯ、Ｓ、ＣＯＯ、ＯＣＯ、ＣＯＳ、ＳＣＯ、ＣＯＮＲ４、ＮＲ４ＣＯ、ＮＨＣＯＯ、ＯＣＯＮＨ、ＮＨＣＯＳ、ＳＣＯＮＨ、ＮＨＣＯＮＨのいずれか（ここでＲ４は、水素または炭素数１〜１８のアルキル基またはアルケニル基）があげられる。
【００４１】
本発明のＮ−アシルアミン化合物は任意の方法で製造できる。一般式（１）に示されるように−ＣＯＮＲ_２−のアミド結合を必須としていることから、本発明のＮ−アシルアミン化合物は、Ｒ１を含むカルボン酸と、Ｒ_２及びＲ_３を含むアミノ化合物と、生体由来物質Ｘを出発原料とするのが合成しやすさの点で最も好ましい。アミノ化合物は生体由来物質Ｘを結合するという点でアミノ基以外にもう一つの反応性官能基を有していることが好ましい。反応性官能基の具体例として、アミノ基、水酸基、カルボキシル基、チオール基、アルデヒド基、イソシアナート基、グリシジル基などが挙げられる。
【００４２】
それぞれの出発原料を結合する順番については特に制限はなく、生体由来物質ＸをＡで表される結合でアミノ化合物に結合した後にカルボン酸と反応させてもよく、初めにカルボン酸とアミノ化合物を結合して中間体アミド化合物を得た後にＸを結合してもよい。アミド結合−ＣＯＮＲ_２−を含む中間体アミド化合物は、Ｒ_１を含むカルボン酸あるいはその誘導体と、Ｒ_２及びＲ_３を含むアミノ化合物との反応によって得られる。誘導体とはカルボン酸塩化物、カルボン酸臭化物、カルボン酸ヨウ化物、カルボン酸エステル、カルボン酸無水物、カルボン酸塩を指す。中間体アミド化合物は、カルボン酸をジシクロヘキシルカルボジイミドなどのカルボジイミドで活性化してアミノ化合物を添加し、副産物として生成するジシクロヘキシルウレアなどの尿素化合物を除去した後に溶媒を除去することによって得られるほか、カルボジイミドで活性化した後に、Ｎ−ヒドロキシスクシンイミドでカルボキシル基を更に活性化してからアミノ化合物を加える方法でも得られる。また、ピリジン、トリエチルアミン、ジメチルアミノピリジンなどの触媒存在下にカルボン酸塩化物あるいはカルボン酸臭化物あるいはカルボン酸ヨウ化物あるいはカルボン酸無水物にアミノ化合物を添加し、触媒の塩を除去した後に溶媒を除去することによっても得られる。更に、カルボン酸エステルにアミノ化合物を添加し、溶媒を除去することによっても得られる。アミノ化合物のアミノ基以外の反応性官能基が、ｔｅｒｔ−ブトキシカルボニル基、カルボベンゾキシ基、ニトロフェニル基、ホルミル基、アシル基、ハロアシル基、ベンゾイル基、トシル基などの保護基によって保護されている場合は、アミド結合反応を終了した後に保護基をはずすことが好ましい。
【００４３】
アミノ化合物あるいは中間体アミド化合物と生体由来物質Ｘを結合する方法は特に限定されず適宜選択できる。
【００４４】
Ｘとして単糖、オリゴ糖、多糖に代表される糖をエーテル結合、チオエーテル結合で結合する場合、一般的に、まず糖の水酸基をアシル基、ハロアシル基、ベンジリデン基、ｉｓｏ−プロピリデン基などで保護する。保護化の反応については畑中らの「糖質の科学と工学」（講談社）に詳細に記載されている。保護・脱保護の容易さから、アセチル基あるいはハロアセチル基が好ましく用いられる。保護した糖をクロロホルム、ジクロロエタンなどの溶媒に溶解し、四塩化すずあるいは三フッ化硼素などのルイス酸触媒存在下に、水酸基あるいはチオール基を有する中間体アミド化合物またはアミノ基を保護された出発原料アミノ化合物を添加して反応させた後、触媒と溶媒を除去して得られる生成物をナトリウムメトキシドなどのルイス塩基で処理して保護基をはずすと糖で修飾されたＮ−アシルアミン化合物を得る。この反応によって得られる糖修飾Ｎ−アシルアミン化合物において、糖は還元末端のアノマー位の炭素で結合している。また、ルイス酸触媒の代わりに臭化水素でアノマー位の炭素を臭素化した後に、過塩素酸銀存在下に中間体アミド化合物またはアミノ基を保護された出発原料アミノ化合物と反応させても同じ糖修飾Ｎ−アシルアミン化合物が得られる。
【００４５】
糖をアミド結合で結合する一般的な方法として、カルボキシル基を有する中間体アミド化合物またはアミノ基を保護された出発原料アミノ化合物をジシクロヘキシルカルボジイミドなどのカルボジイミドで活性化して、ガラクトサミン、キトサンなどのアミノ基を含む糖を添加し、副産物として生成するジシクロヘキシルウレアなどの尿素化合物を除去した後に溶媒を除去することによって得られるほか、カルボジイミドで活性化した後に、Ｎ−ヒドロキシスクシンイミドでカルボキシル基を更に活性化してから糖を加える方法でも得られる。グルコン酸、グルクロン酸などのように糖にカルボキシル基が含まれる場合には、水酸基を保護した後、糖をカルボジイミドで活性化して、アミノ基を有する中間体アミド化合物またはアミノ基を保護された出発原料アミノ化合物を添加し、副産物として生成するジシクロヘキシルウレアなどの尿素化合物を除去した後に溶媒を除去することによって得られるほか、カルボジイミドで活性化した後に、Ｎ−ヒドロキシスクシンイミドでカルボキシル基を更に活性化してから糖を加える方法でも得られる。
【００４６】
糖をエステル結合またはチオエステル結合で結合する一般的な方法としては、グルコン酸、グルクロン酸などのように糖にカルボキシル基が含まれる場合には、ベンジル基などで水酸基を保護した後、糖を塩化チオニル、三塩化リン、五塩化リンなどで処理してカルボキシル基を塩素化した後、ピリジン、トリエチルアミン、ジメチルアミノピリジンなどの触媒存在下に、水酸基またはチオール基を含む中間体アミド化合物またはアミノ基を保護された出発原料アミノ化合物を添加し、触媒の塩と溶媒を除去することによって得られる。
【００４７】
カルボキシル基を含む中間体アミド化合物またはアミノ基を保護された出発原料アミノ化合物のカルボキシル基を予め酸塩化物、酸臭化物、酸ヨウ化物、酸無水物に変換した化合物を、ピリジン、トリエチルアミン、ジメチルアミノピリジンなどの触媒存在下に糖に添加し、触媒の塩と溶媒を除去することによっても得られる。
【００４８】
Ｘとしてアミノ酸、オリゴペプチド、ポリペプチド、タンパク質、タンパク質のフラグメントに代表されるペプチド化合物をアミド結合で結合する場合、一般的に、カルボキシル基を有する中間体アミド化合物またはアミノ基を保護された出発原料アミノ化合物をカルボジイミドで活性化して、アミノ基を含むアミノ酸またはポリペプチドなどを添加し、副産物として生成するジシクロヘキシルウレアなどの尿素化合物を除去した後に溶媒を除去することによって得られるほか、カルボジイミドで活性化した後に、Ｎ−ヒドロキシスクシンイミドでカルボキシル基を更に活性化してからペプチド化合物を加える方法でも得られる。この方法を用いるとペプチド化合物のアミノ末端のアミノ基またはリジン残基のアミノ基で結合できる。また、ペプチド化合物をエステル結合またはチオエステル結合で結合する場合、一般的に、カルボキシル基を有する中間体アミド化合物またはアミノ基を保護された出発原料アミノ化合物を塩化チオニル、三塩化リン、五塩化リンなどで処理してカルボキシル基を塩素化した後、ピリジン、トリエチルアミン、ジメチルアミノピリジンなどの触媒存在下に、水酸基またはチオール基を含むアミノ化合物を添加し、触媒の塩と溶媒を除去することによって得られる。この方法を用いるとペプチド化合物のセリン残基またはスレオニン残基の水酸基、システイン残基のチオール基で結合できる。また、カルボキシル基を有するペプチド化合物を塩化チオニル、三塩化リン、五塩化リンなどで処理してカルボキシル基を塩素化した後、ピリジン、トリエチルアミン、ジメチルアミノピリジンなどの触媒存在下に、水酸基またはチオール基を含む中間体アミド化合物またはアミノ基を保護された出発原料アミノ化合物を添加し、触媒の塩と溶媒を除去することによっても得られる。この方法を用いるとペプチド化合物のカルボキシ末端のカルボキシル基、アスパラギン酸残基またはグルタミン酸残基のカルボキシル基で結合できる。
このときペプチド化合物にアミノ基が含まれている場合には、保護基で保護されていることが好ましく、保護基は結合反応終了後はずすことができる。
【００４９】
溶媒、反応温度、反応時間、精製方法は原料や前駆体物質の化学構造や反応の種類によって適宜選択できる。
【００５０】
本発明でいう生分解性とは生体内で分解される性質を意味し、酵素が関与するか否かにかかわらず主として加水分解反応によって分解し、生体適合性のある物質を生成することである。生体適合性のある物質とは、もともと生体内にある物質と化学構造が同じであるか、酵素、免疫、肝機能、腎機能などにより代謝・排泄されうる物質である。本発明のＮ−アシルアミン化合物が分解されて生じた複数の構成物が生体由来の物質であれば、それらは生体適合性物質であり、すなわち、そのＮ−アシルアミン化合物は生分解性である。
【００５１】
本発明のＮ−アシルアミン化合物は表面修飾分子として、少なくとも表面に存在する形態に加工して好ましく用いられる。表面に存在する形態に加工するためには、本発明のＮ−アシルアミン化合物を基材粒子の表面にコーティングして用いてもよく、リポソームまたはミセル、すなわちＮ−アシルアミン化合物自体で粒子形成して用いてもよい。この場合、使用するＮ−アシルアミン化合物の種類は、目的に応じて適宜調節でき限定されない。１種類のＮ−アシルアミン化合物のみを用いてもよいし、複数のＮ−アシルアミン化合物を混合して用いてもよい。ここでいうリポソームとは、Ｎ−アシルアミン化合物がＲ_１の部分で会合して二重膜を形成し、その二重膜が球状になった組成物である。また、ミセルとはＮ−アシルアミン化合物がＲ_１の部分で会合してコアを形成し、Ｘが表面に分布した球状の組成物である。
【００５２】
この場合に、リポソームまたはミセルや粒子の形状は特に限定されるものではない。大きさについては特に限定されず、目的に合った大きさを選択できる。細胞の間隙のサイズは組織によって異なり、細胞が取り込めるサイズも細胞により様々である。サイズが大きすぎると組織や細胞内に取り込めなくなり、一方、サイズが小さすぎると目的以外の組織や細胞にも取り込まれやすくなる。また、組織や細胞の表面に粒子を吸着させる目的のためには、組織や細胞に取り込まれないサイズがよい。平均粒径として０．１ｎｍ〜５ｍｍが好ましい。組織や細胞に取り込ませる目的の場合には、０．１ｎｍ〜５００μｍが好ましく、特に肝臓や肝実質細胞に取り込ませる目的の場合には１０ｎｍ〜５００ｎｍがより好ましい。粒子の平均粒径は走査型電子顕微鏡Ｓ−２４００（日立社製）を用いて複数の粒子の粒径を測定し、その平均値を算出することによって得られる。またレーザー散乱型粒度分布計（Ｍｉｃｒｏｔｒａｃ ＡＳＶＲ／Ｍｉｃｒｏｔｒａｃ−ＨＲＡ（９３２０−Ｘ１００））を用いると直接平均粒径を知ることができる。
【００５３】
本発明のＮ−アシルアミン化合物を基材粒子の表面にコーティングして用いる場合、粒子を構成する基材は特に限定されないが、酵素、免疫、肝機能、腎機能などにより分解・代謝・排泄される物質からなることが好ましい。このような物質の具体例としては、ポリエチレングリコール、ポリ乳酸、ポリグリコール酸、ポリカプロラクトン、ポリビニルアルコール、セルロース、アガロース、デンプン、アルギン酸、キチン、キトサン、およびこれらの物質の誘導体、共重合体などが挙げられるが、これらに限定されるものではない。細胞に取り込ませる目的で使用する場合には生分解性であることが好ましく、ポリ乳酸、ポリグリコール酸、ポリカプロラクトンおよびこれらの共重合体が好ましく用いられる。粒子基材の生分解性の測定には公知の方法を用いることができる。例えば、材料を水または塩溶液に適当な固液比で浸漬し、適当な温度で適当な期間振とうまたは攪拌する方法や、生体内に近い条件で測定する場合には生理食塩水またはリン酸緩衝生理食塩水に浸漬して３７℃で振とうまたは攪拌する方法が好ましく用いられる。材料の乾燥重量、体積、平均分子量、外観などを天秤、ゲル浸透クロマトグラフィー、走査型電子顕微鏡などを用いて測定し、経時変化を調べることによって生分解性能を評価する。
【００５４】
本発明のＮ−アシルアミン化合物の粒子表面上における密度は、高すぎるとリガンドの配向が損なわれるだけでなく、立体的な障害が生じ、細胞上に存在するレセプターによって認識されにくくなる。一方、低すぎるとリガンドとレセプターの接触機会が減少するため細胞によって認識されにくくなる。したがって、粒子表面の表面密度としては、１〜１０００ｎｍ^２に１分子が好ましい。
【００５５】
本発明のＮ−アシルアミン化合物をリポソームとして、あるいは基材粒子の表面にコーティングして用いる場合には、リポソームあるいは基材粒子の中に生理活性物質、薬剤、遺伝子が単独であるいは複数含まれていてもよい。生理活性物質とは、ＰＤＧＦ、ＶＥＧＦ、ＨＧＦ、ＦＧＦ、ＥＧＦなどの増殖因子、ＩＮＦ、ＴＧＦ、インターロイキンなどのサイトカイン、ホルモンなどが挙げられる。また、ワクチンとして用いられる死菌、毒素、糖鎖、ペプチド類などの抗原も含まれる。薬剤とは、造影剤、抗癌薬、抗炎症薬、抗アレルギー薬、循環器薬、降圧薬、昇圧薬、抗血小板薬、抗凝固薬、向精神薬、疼痛治療薬、抗生物質、消化器薬、泌尿器薬、内分泌薬、蛍光試薬などが挙げられる。遺伝子は、オリゴヌクレオチド、ポリヌクレオチド、ＤＮＡ，ＲＮＡなどが挙げられる。特に肝臓や肝細胞を標的とする場合には、リバビリン、ラミブシン、インターフェロンなどの抗ウイルス剤、シクロスポリンなどの抗炎症剤、シスプラチン、カルボプラチン、５−Ｆｕ、マイトマイシン、シクロフォスファミド、メトトレキセートなどの抗癌剤、イオパミドール、イオヘキソールなどのＸ線造影用造影剤、^１３１Ｉ、^９９ｍＴｃ、^１３１Ｉ−ＨＳＡ、^６７Ｇａ、^３Ｈ、^２４Ｎａ、^８６Ｒｂ、^８７ｍＳｒ、^１８Ｆ−ＦＤＧなどのシンチグラフィー用造影剤、Ｇｄ化合物、Ｆｅ化合物、Ｍｎ化合物などの核磁気共鳴用造影剤、ポリデキストロース、オクタフルオロプロパンなどの超音波造影剤、フィブラートなどの高脂血症薬、ＨＧＦなどの増殖因子、エンドスタチン、アンギオスタチンなどの血管新生阻害因子などが好ましく用いられる。
【００５６】
本発明におけるＮ−アシルアミン化合物によって被覆された粒子の調製方法は特に限定されないが、よく知られたスプレー法、エマルジョン法が好ましく用いられる。本発明でいう水溶性溶媒とは、メタノール、エタノール、アセトン、ＤＭＳＯ、ＤＭＦ、ＤＭＡｃなど水に溶解しやすい溶媒であり、非水溶性溶媒とは、クロロホルム、ジクロロメタン、ジエチルエーテル、酢酸エチルなどの水に不溶な溶媒を指す。
【００５７】
エマルジョン法では、粒子基材をクロロホルム、塩化メチレンなどの非水溶性溶媒に溶解し、本発明のＮ−アシルアミン化合物を溶解した水中に滴下し、攪拌して有機溶媒を蒸発させる。本発明のＮ−アシルアミン化合物の濃度、粒子基材の濃度、攪拌回転数、温度、圧力を適当に選ぶことによって、粒子のサイズを適宜変えられる。攪拌後、蒸留水で洗浄し、遠心分離または濾過により被覆粒子を水から分離して被覆粒子を得る。粒子基材濃度は、高すぎると溶液の操作性が悪いだけでなく、基材粒子溶液がＮ−アシルアミン化合物水溶液中で分散せず、低すぎると得られる粒子が少なく効率が悪いので、０．００１〜３０ｗ／ｖ％が好ましく、更に０．０１〜２０ｗ／ｖ％が特に好ましい。攪拌速度は速すぎると粒径にばらつきが生じ、遅すぎると細かい粒子が得られないので、１００〜１０００回転／分が好ましく、更には１００〜４００回転／分が特に好ましい。温度条件は、粒子基材を溶解する溶媒に依存し、高すぎると溶媒がすぐに蒸発するため細かい粒子が得られにくいほかエマルジョンの分散性が悪くなり、低すぎると溶媒が蒸発せずに粒子調製に時間がかかるので、０℃〜溶媒の沸点が好ましい。溶媒の蒸発を促進するために、減圧で操作することもでき、このときの圧力は１気圧以下であることが好ましい。例えば、粒子基材を０．０１〜２０ｗ／ｖ％の濃度になるようにクロロホルム、塩化メチレンなどの非水溶性溶媒に溶解し、粒子基材溶液の１〜１０００倍容の、本発明のＮ−アシルアミン化合物を０．０１〜１０％の濃度になるように溶解した水中に滴下し、１００〜４００回転／分の速度で適温で攪拌し、遠心分離または濾過により被覆粒子を水から分離することによって、Ｎ−アシルアミン化合物の表面密度が１〜１０００ｎｍ^２／１分子で平均粒径が０．１ｎｍ〜５ｍｍの、Ｎ−アシルアミン化合物でコーティングされた粒子が得られる。
【００５８】
また、本発明のＮ−アシルアミン化合物と粒子基材を濃度がそれぞれ０．０１〜１０％、０．００１〜１０％になるようにクロロホルム、塩化メチレンなどの非水溶性溶媒に溶解し、その溶液を粒子基材溶液の１０〜１０００倍容の水中に滴下して、好ましくは１００〜１０００回転／分、更に好ましくは１００〜４００回転／分の速度で適温で攪拌し、遠心分離または濾過により被覆粒子を水から分離することによっても被覆粒子が得られる。
【００５９】
超音波で処理しながら攪拌してもよく、攪拌せずに超音波処理のみ行うのでもよい。例えば、ＵＳ−５０プローブ型超音波装置（Ｎｉｓｓｅｉ）を用いる場合は、出力が低すぎると水中で粒子基材溶液が分散せず、高すぎると平均粒径が小さくなるため、１０〜１００Ｗの出力が好ましい。
【００６０】
また、本発明のＮ−アシルアミン化合物と粒子基材とを、好ましくは０．００１〜３０ｗ／ｖ％の濃度でＤＭＳＯ、ＤＭＦ、メタノール、エタノール、アセトンなどの水溶性溶媒に溶解した後に溶液を透析チューブに入れて水中で透析した後、透析チューブ内の懸濁液を遠心分離または濾過により被覆粒子を水から分離することによってＮ−アシルアミン化合物の表面密度が１〜１０００ｎｍ^２／１分子で平均粒径が０．１ｎｍ〜５ｍｍの被覆粒子を得る。
【００６１】
リポソームまたはミセルを調製する場合には、本発明のＮ−アシルアミン化合物を好ましくは０．００１〜３０ｗ／ｖ％の濃度で直接水に溶解してもよいし、ＤＭＳＯ、ＤＭＦ、メタノール、エタノール、アセトンなど水溶性溶媒に好ましくは０．００１〜３０ｗ／ｖ％の濃度で溶解した後に粒子基材溶液の１〜１０００倍容の水中に滴下してもよい。好ましくは水中に滴下した後に超音波処理することによって粒径のそろったリポソームまたはミセルが得られる。遠心分離または濾過により被覆粒子を水から分離することによってリポソームまたはミセルが得られる。
【００６２】
生理活性物質や薬剤を粒子やリポソームまたはミセルに担持させたい場合には、粒子基材の溶液中に生理活性物質や薬剤あるいはそれらの溶液を共存させて後に上記の操作を行う。生理活性物質や薬剤の粒子中における濃度は、仕込み濃度等を適宜選択することができ、使用目的の濃度に合わせて変えることができる。例えばエマルジョン法では、生理活性物質や薬剤が疎水性の場合には、粒子基材と薬剤をクロロホルム、塩化メチレンなどの非水溶性溶媒に溶解・混合し、本発明のＮ−アシルアミン化合物を０．０１〜１０％の濃度になるように溶解した水中に滴下し、１００〜４００回転／分の速度で適温で攪拌することによって得られる。生理活性物質や薬剤が親水性の場合には、薬剤の水溶液と、粒子基材をクロロホルム、塩化メチレンなどの非水溶性溶媒に溶解した溶液とを攪拌・混合してエマルジョンを調製し、そのエマルジョンを本発明のＮ−アシルアミン化合物を０．０１〜１０％の濃度になるように溶解した水中に滴下し、１００〜４００回転／分の速度で適温で攪拌することによって得られる。生理活性物質や薬剤の濃度は、目的の薬効を発揮するために必要な濃度に自由に調節できる。また、エマルジョンを作成する場合、ポリエチレングリコールやポリビニルアルコールなどの分散剤を用いてもよい。構造によっては、Ｎ−アシルアミン化合物自身が分散剤の役目を果たすことがあるのでポリエチレングリコールなどの分散剤の代わりに、または分散剤と共にＮ−アシルアミン化合物をエマルジョン調製時に使用してもよい。生理活性物質や薬剤を含有させる際にもＮ−アシルアミン化合物の濃度、粒子基材の濃度、攪拌回転数、温度を適当に選ぶことによって、粒子のサイズを適宜変えられる。攪拌後、蒸留水で洗浄した後、遠心分離または濾過により被覆粒子を水から分離することによって粒子を得る。粒子基材濃度は、高すぎると溶液の操作性が悪いだけでなく、基材粒子溶液がＮ−アシルアミン化合物水溶液中で分散せず、低すぎると得られる粒子が少なく効率が悪いので、０．００１〜３０ｗ／ｖ％が好ましく、更に０．０１〜２０ｗ／ｖ％が特に好ましい。攪拌速度は速すぎると粒径にばらつきが生じ、遅すぎると細かい粒子が得られないので、１００〜１０００回転／分が好ましく、更には１００〜４００回転／分が特に好ましい。温度条件は、粒子基材を溶解する溶媒に依存し、高すぎると溶媒がすぐに蒸発するため細かい粒子が得られにくいほかエマルジョンの分散性が悪くなり、低すぎると溶媒が蒸発せずに粒子調製に時間がかかるので、０℃〜溶媒の沸点が好ましい。
【００６３】
生理活性物質や薬剤を含有したリポソームまたはミセルを調製する場合には、本発明のＮ−アシルアミン化合物を好ましくは０．００１〜３０ｗ／ｖ％の濃度でＤＭＳＯ、ＤＭＦ、メタノール、エタノール、アセトンなど水溶性溶媒に溶解し、薬剤を添加し混合した後に粒子基材溶液の１〜１０００倍容の水中に滴下してもよい。好ましくは水中に滴下した後に超音波処理することによって粒径のそろったリポソームまたはミセルが得られる。
【００６４】
本発明における医療用器材とは病気を診断、治療、予防するために用いる器材であり、具体的には生理活性物質、薬物、遺伝子等を含有したキャリアやベクター、ワクチンなどのドラッグデリバリーシステム、カテーテル類、人工血管や人工骨頭のようなインプラントデバイス、人工心肺、人工透析器のような血液回路、吸着材のような体外循環システム用基材、マイクロプレートや粒子などの検査診断薬用基材、細胞足場材や細胞キャリアなどの細胞治療・再生医療システム用基材、クロマトグラフィー担体などを指すがこれに限定されるものではない。
【００６５】
これらの医療用器材の表面にＮ−アシルアミン化合物を配設することによって、生体適合性が著しく向上する。例えば、カテーテル、血液回路は血液と接触することによって血小板や凝固カスケードが活性化し血栓を形成するが、例えば抗血栓性物質であるヘパリンを結合したＮ−アシルアミン化合物を表面に被覆することによってこのような基材表面での血栓形成反応を抑えることができる。また、インプラントデバイスや細胞治療・再生医療システム用基材では、基材表面が異物反応、炎症反応を誘発する、例えばアルギニン−グリシン−アスパラギン酸のアミノ酸配列を含むペプチドやコンドロイチン硫酸を結合したＮ−アシルアミン化合物を表面に被覆することによって、目的部位の組織や細胞との親和性、接着性が向上する。また、検査診断薬や体外循環治療においては、定量したい物質、体外に除去したい物質と相互作用する物質を結合したＮ−アシルアミン化合物を粒子や繊維にコーティングすることによって、それらの物質を定量、除去できる。
【００６６】
これらの医療用器材表面にＮ−アシルアミン化合物を付与する方法は特に限定されず、Ｎ−アシルアミン化合物の溶液に医療器材を浸漬した後、乾燥して溶媒を除去することによって得ることもできるし、また、医療用器材の原料とＮ−アシルアミン化合物を混合した後に成型することもできる。
【００６７】
以下に実施例を示すが、本発明はこれら実施例により限定されるものでない。
【００６８】
【実施例】
＜合成例１＞
Ｄ−乳糖２０ｇをピリジン３２０ｍｌに懸濁後、氷浴中で攪拌しながら無水酢酸３２０ｍｌを滴下した。３日間室温で攪拌後、反応溶液を７００ｍｌの水中に投入した。沈殿を濾別した後、ｐＨ７になるまで水洗した。減圧で乾燥後、産物２６ｇをジエチルエーテル７０ｍｌと酢酸エチル２０ｍｌの混合溶媒に溶解し、硫酸ナトリウムを加えて攪拌後、硫酸ナトリウムを濾別し、溶媒を減圧除去してアセチル化乳糖２７ｇを得た。
【００６９】
リノール酸１０ｇとＮ−ヒドロキシスクシンイミド４．９ｇをジクロロメタンに溶解した後、ジシクロヘキシルカルボジイミド８．８ｇと触媒量のピリジンを添加して室温で３日間攪拌した。ジシクロヘキシルウレアを濾別したのち、溶媒を減圧除去して活性化リノール酸１７ｇを得た。全量を再度ジクロロメタンに溶解した後、２−アミノエタノール ３ｍｌを添加し、室温で２日間攪拌した。沈殿を濾別した後、溶媒を減圧除去し、得られた産物を水洗・乾燥してリノール酸アミドを得た。
【００７０】
アセチル化乳糖２ｇとリノール酸アミド３．８ｇをジクロロエタン７０ｍｌに溶解した後、合成ゼオライト粉末２ｇを添加し、窒素ガス雰囲気化に７０℃で攪拌した。三フッ化硼素のジエチルエーテル溶液１ｍｌを添加した後、同条件で５時間攪拌した。ハイフロ・スーパーセルで固相を濾別し、溶媒を減圧除去した。エーテルに溶解後、２−プロパノール中に投与して再沈し沈殿を濾別し減圧乾燥してアセチル化乳糖−リノール酸アミド複合体を得た。
【００７１】
アセチル化乳糖−リノール酸アミド複合体２ｇをメタノール２００ｍｌに溶解した後、ナトリウムメトキシド６００ｍｇを添加して、室温で６時間攪拌した。陽イオン交換樹脂（アンバーライトＩＲ−１２０Ｂ）３５ｇ加えて攪拌したのち、樹脂を濾別して、溶媒を減圧除去して乳糖−リノール酸アミド複合体１．６ｇを得た。
【００７２】
＜比較例１＞
ポリ乳酸（和光純薬；平均分子量５０００）０．６ｇをジクロロメタン６ｍｌに溶解したポリ乳酸溶液を、ポリビニルアルコール（ＰＶＡ）（ナカライテスク；重合度５００）０．３３ｇを溶解した水溶液４００ｍｌに滴下し、室温で８時間攪拌してポリ乳酸粒子を得た。
【００７３】
ポリ乳酸粒子に１ｍｇ／ｍｌのＰＶＬＡ（生化学工業；平均分子量：５００００）のＰＢＳ溶液、またはＰＢＳをそれぞれ添加し、室温で１時間５０分静置した。ＰＢＳ０．５ｍｌで粒子を洗浄した後、０．１ｍｇ／ｍｌＦＩＴＣ標識ＲＣＡ１２０（ホーネンコーポレーション）のＰＢＳ溶液０．２ｍｌを添加し、室温で１時間静置した。ＰＢＳ０．５ｍｌで粒子を洗浄した後、走査型レーザー生物顕微鏡（ＦＬＯＶＩＥＷ；オリンパス）にて粒子表面の蛍光を観察した。レーザー顕微鏡の設定条件は下記の通りであった。
【００７４】

その結果、ｐｏｌｙ−（Ｎ−ｐ−ｖｉｎｙｌ ｂｅｎｚｙｌ−Ｏ−β−Ｄ−ｇａｌａｃｔｏｐｙｒａｎｏｓｙｌ−［１→４］−Ｄ−ｇｌｕｃｏｎａｍｉｄｅ（ＰＶＬＡ）修飾粒子の表面には蛍光が観察されたが、ＰＶＬＡ修飾されていない粒子の表面には蛍光が観察されなかった（図１）。
【００７５】
＜実施例１＞
ポリ乳酸粒子に乳糖−リノール酸アミド複合体のＰＢＳ溶液を添加し、室温で１時間５０分静置した。ＰＢＳ０．５ｍｌで粒子を洗浄した後、０．１ｍｇ／ｍｌＦＩＴＣ標識ＲＣＡ１２０（ホーネンコーポレーション）のＰＢＳ溶液０．２ｍｌを添加し、室温で１時間静置した。ＰＢＳ０．５ｍｌで粒子を洗浄した後、走査型レーザー生物顕微鏡（ＦＬＯＶＩＥＷ；オリンパス）にて比較例１記載の設定条件で粒子表面の蛍光を観察した結果、粒子表面に比較例１のＰＶＬＡ修飾粒子と同程度の蛍光が観察された（図１）。
【００７６】
＜比較例２＞
ポリ乳酸（和光純薬；分子量２００００）１０ｍｇをジクロロメタン０．１ｍｌに溶解したポリ乳酸溶液を、ＰＶＬＡ（生化学工業）またはＰＶＡ（ナカライテスク；重合度５００）０．５ｍｇを溶解した水溶液０．５ｍｌにパスツールピペットで１滴滴下し、室温で１日攪拌して乳糖被覆ポリ乳酸粒子を得た。
【００７７】
ポリ乳酸粒子をＰＢＳ０．５ｍｌで洗浄した後、０．１ｍｇ／ｍｌＦＩＴＣ標識ＲＣＡ１２０（ホーネンコーポレーション）のＰＢＳ溶液０．２ｍｌを添加し、室温で１時間静置した。ＰＢＳ０．５ｍｌで粒子を洗浄した後、比較例１記載の設定条件で走査型レーザー生物顕微鏡（ＦＬＯＶＩＥＷ；オリンパス）にて粒子表面の蛍光を観察した結果、蛍光が観察された（図２）。蛍光発色部分中の任意の３平面の平均蛍光強度は、ＰＶＬＡで被覆した粒子は４０７３±２１、ＰＶＡで被覆した粒子は１５７±４であった。
【００７８】
＜実施例２＞
ポリ乳酸（和光純薬；分子量２００００）１０ｍｇをジクロロメタン０．１ｍｌに溶解したポリ乳酸溶液を、乳糖−リノール酸アミド複合体１ｍｇを溶解した水溶液０．５ｍｌにパスツールピペットで数滴滴下し、室温で１日間攪拌して乳糖被覆ポリ乳酸粒子を得た。
【００７９】
ポリ乳酸粒子をＰＢＳ０．５ｍｌで洗浄した後、０．１ｍｇ／ｍｌＦＩＴＣ標識ＲＣＡ１２０（ホーネンコーポレーション）のＰＢＳ溶液０．２ｍｌを添加し、室温で１時間静置した。ＰＢＳ０．５ｍｌで粒子を洗浄した後、比較例１記載の設定条件で走査型レーザー生物顕微鏡（ＦＬＯＶＩＥＷ；オリンパス）にて粒子表面の蛍光を観察した結果、粒子表面に比較例２のＰＶＬＡ修飾粒子と同程度の蛍光が観察された（図２）。蛍光発色部分中の任意の３平面の平均蛍光強度は３９４１±５６であった。
【００８０】
＜比較例３＞
細胞を低温でホモジュナイザーで処理し、リン酸緩衝生理食塩水に懸濁した後、超遠心して不溶物を除去して得られた細胞抽出液を調製した。
【００８１】
５ｍｇのＰＶＬＡを０．１ｍｌのメタノールに溶解し、１０μｇ／ｍｌのフルオレセイン２０μｌを添加した後、この溶液を０．９ｍｌのリン酸緩衝生理食塩水中に添加した。細胞抽出液を添加し、３７℃で２４時間振とうした。反応液を透析膜に入れて蒸留水に対して透析した。チューブ内の液を凍結乾燥後、高速液体クロマトグラフィーで分子量解析を行った結果、細胞抽出液処理前後でＰＶＬＡの分子量は約５００００で変化がなく細胞内酵素によって分解されなかった。
【００８２】
＜実施例３＞
細胞を低温でホモジュナイザーで処理し、リン酸緩衝生理食塩水に懸濁した後、超遠心して不溶物を除去して得られた細胞抽出液を調製した。
【００８３】
５ｍｇの乳糖−リノール酸アミド複合体を０．１ｍｌのメタノールに溶解し、１０μｇ／ｍｌのフルオレセイン２０μｌを添加した後、この溶液を０．９ｍｌのリン酸緩衝生理食塩水中に添加した。細胞抽出液を添加し、３７℃で蛍光強度の時間的変化を蛍光分光光度計（Ｆ−２０００；日立製作所）を用いて下記の条件で測定した。
【００８４】
励起波長：４９０ｎｍ
スキャン速度：６０ｎｍ／分
レスポンス：２秒
バンドパス：励起側１０ｎｍ、蛍光側１０ｎｍ
光電子倍増管：４００Ｖ
蛍光強度が時間と共に１６０８→６→３０と変化し、リノール酸アミドおよびリノール酸にフルオレセインを添加した場合と同等レベルになり、細胞内酵素によって乳糖−リノール酸アミド複合体が分解されることが証明された。
【００８５】
【発明の効果】
本発明の表面被覆化合物は体内で分解するため、本化合物で被覆することによって、目的の組織や細胞に安全に薬物等を配送できる。
【図面の簡単な説明】
【図１】実施例のＰＶＬＡおよび乳糖−リノール酸複合体で被覆した粒子のレーザー顕微鏡写真である。
【図２】実施例のＰＶＬＡおよび乳糖−リノール酸複合体で被覆した粒子のレーザー顕微鏡写真である。
【図３】実施例の乳糖−リノール酸複合体にフルオレセインと細胞抽出液を添加した場合の蛍光変化である。[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to an N-acylamine compound for delivering a physiologically active substance, a drug, a gene, and the like to a target place, and a medical carrier using the same.
[0002]
[Prior art]
Delivering a drug, a physiologically active substance, a gene, or the like administered to a living body to only a target site of action is intended to prevent side effects induced by these substances and to enhance the efficacy of these substances. Is also an important issue.
[0003]
One method of solving this problem is to directly modify a drug or the like with a ligand having a high affinity for the cell surface substance of the organ or tissue at the site of action, or to carry the drug or the like on a carrier modified with the ligand. It is. Specific examples of the ligand include asialoglycoprotein and galactose having affinity for hepatocytes, sialyl Lewis X having affinity for vascular endothelial cells, transferrin having affinity for cancer cells, and HIV virus having affinity for leukocytes. gp120, tuftsin, mannose-6-phosphate. Specific examples of the carrier include fine particles and liposomes. Many researches and developments have been made on drug carriers modified with ligands (for example, see Non-Patent Document 1), and it is reported that drugs and the like can be delivered into isolated cells at the test tube level. However, very few cases have been successful in animal experiments, and none has reached clinical application. In vivo, before these drugs reach the target location, their number is reduced by immunity or phagocytic phagocytic action before reaching the target location, or even if the target location is reached, the ligand density is insufficient. In addition, the chance of contact with the receptor on the surface of the target cell was small, and the adhesion was not reached. For example, Wu et al. Prepared a liposome for delivering a drug to the liver using asialofetuin having N-acetylgalactosamine at the end as a ligand (for example, see Non-Patent Document 2). However, asialofetuin elicits an immune response due to its protein portion. In addition, even if the amphipathic substance that constitutes the liposome is arranged on the particle surface, the surface density is low due to the lack of a chemical structure that produces strong interaction between the amphipathic substances on the surface, and the cell density is low. It is hard to be recognized. Further, if the interaction is weak, the amphipathic substance is easily detached from the liposome due to the interaction with the lipoprotein in the serum or the lipid in the tissue, which also reduces the interaction with the cell (for example, Non-Patent Document 3). reference).
[0004]
In order to solve this problem, Kobayashi et al. Reported a compound (poly- (Np-vinyl benzyl-O-β-D-galactopyranosyl) obtained by immobilizing galactose which is a ligand of an asialoglycoprotein receptor of hepatocytes on polystyrene. -[1 → 4] -D-gluconamide (PVLA) was reported (for example, see Non-Patent Document 4), since this polymer has a saccharide moiety that is hydrophilic and has a saccharide for each repeating unit. Watanabe et al. Reported that particles were not phagocytosed, but were accumulated specifically in the liver without being phagocytosed, and were loaded with certain drugs to prevent acute phagocytosis. It has been reported that when administered to hepatitis model animals, a life-saving effect is obtained (see, for example, Non-Patent Document 5). Over the due to the use of non-degradable polystyrene, indefinitely remain in the cell without being degraded.
[0005]
In addition, amphiphilic substances bound to sugars which are considered to be able to be arranged on the particle surface have been developed as biodegradable surfactants (for example, see Non-Patent Documents 6-9). However, it is unclear whether such surfactants can be degraded by microorganisms in the global environment, and whether they are degraded in animals, especially in humans and cells.
[0006]
It is desirable that the modified substance that has reached the target site and has completed its role is excreted as it is, undergoes decomposition / metabolism, or is absorbed. Especially when the cell surface receptor for the ligand induces endocytosis, or when it is desired to exert the effect of the drug in the cell, the carrier enters the cell together with the drug and is degraded by enzymes in the cell. It is desirable to be done.
[0007]
As means for solving this problem, many methods using a ligand-modifying substance or a ligand-binding carrier based on a biodegradable substance such as polyester, polyamino acid, lipid, etc. have been reported (for example, Patent Documents 1-7, Non-Patent Document 1). Patent Document 10). However, among these existing substances, the binding between the ligand and the biodegradable substance is non-degradable, or elicits an immune response due to the presence of the polypeptide structure. There was no ingenuity in the chemical structure that would cause it.
[0008]
[Patent Document 1]
U.S. Pat. No. 6,210,717
[0009]
[Patent Document 2]
US Patent No. 6410057
[0010]
[Patent Document 3]
US Pat. No. 6,254,890
[0011]
[Patent Document 4]
U.S. Pat. No. 6,267,978
[0012]
[Patent Document 5]
U.S. Pat. No. 5,543,158
[0013]
[Patent Document 6]
JP-A-11-269097 (page 1-27)
[0014]
[Patent Document 7]
JP-A-5-194199 (pages 1-9)
[0015]
[Non-patent document 1]
Vyas SP et al., Advanced Drug Delivery Reviews, 2000; 43: 101-164.
[0016]
[Non-patent document 2]
Wu J et al., Hepatology. 1998; 27: 772-778.
[0017]
[Non-Patent Document 3]
(Spanjer HH et al.) Biochemica et Biophysica Acta, 1983; 734: 40-47.
[0018]
[Non-patent document 4]
Kobayashi K et al., Polymer Journal (Polm J). 1985; 17: 567.
[0019]
[Non-Patent Document 5]
Watanabe Y et al., Journal of Biomaterials Science Polymer Edition (J Biomater Sci Polymer Edn). 2000; 11: 833-848.
[0020]
[Non-Patent Document 6]
Soderman O et al., Current Opinion in Colloid and Interface Science (Curr Opin Colloid Interface Sci). 2000; 4: 391-401
[0021]
[Non-Patent Document 7]
Holmberg K et al., Current Opinion in Colloid and Interface Science (Curr Opin Colloid Interface Sci). 2001; 6: 148-159.
[0022]
[Non-Patent Document 8]
Stubenrauch C et al., Current Opinion in Colloid and Interface Science (Curr Opin Colloid Interface Sci). 2001; 6: 160-170.
[0023]
[Non-Patent Document 9]
Kawakami S et al. Biochimica et Biophysica Acta, 2000; 1524: 258-256.
[0024]
[Non-Patent Document 10]
Chenvier P et al., Chemical Communications (Chem Comm), 2002; 20: 2446-2447.
[0025]
[Problems to be solved by the invention]
Delivering a drug or the like administered to a living body only to a target site of action is important from the viewpoint of preventing side effects induced by these substances and enhancing the efficacy. Has a chemical structure that has a hydrophilic portion that can also avoid immunity and phagocytosis, has a chemical structure that can achieve a ligand surface density that increases the chance of contact with the cell surface receptor at the site of action, and is degraded even in cells No safe surface modifier has been realized so far.
[0026]
In view of the problems of the conventional technology, the present inventors have studied various ligand binding compounds. As a result, a compound obtained by binding a ligand and a fatty acid via an amine compound has a degradable, high-density structure. The present invention has been found, and the present invention has been achieved. That is, an object of the present invention is to provide a surface modifying substance for delivering a drug or the like to a target site and a method for producing the same.
[0027]
[Means for Solving the Problems]
The present invention mainly has the following configurations in order to achieve the above object.
"(1) The following general formula (1)
[0028]
Embedded image

[0029]
(Where R ₁ Is an alkyl group having 1 to 23 carbon atoms or an alkenyl group having at least one double bond, R ₂ Is hydrogen or an alkyl or alkenyl group having 1 to 18 carbon atoms, R3 is an alkylene group having 1 to 18 carbon atoms, A is O, S, COO, OCO, COS, SCO, CONR4, NR4CO, NHCOO, OCONH, NHCOS, An N-acylamine compound represented by SCONH or NHCONH (where R4 is hydrogen or an alkyl or alkenyl group having 1 to 18 carbon atoms) and X is a biological substance.
(2) R in the general formula (1) ₂ Is hydrogen. (1) The N-acylamine compound according to (1),
(3) R in the general formula (1) ₁ (1) or (2), wherein the alkyl group or alkenyl group is a linear group having 12 to 23 carbon atoms.
(4) R in the general formula (1) ₁ Is an alkenyl group, and the number of double bonds is 1 to 5, The N-acylamine compound according to any one of (1) to (3),
(5) The N-acylamine compound according to (4), wherein R3 in the general formula (1) is an alkylene group having 1 to 4 carbon atoms.
(6) The N-acylamine compound according to any one of (1) to (5), wherein X in the general formula (1) is selected from sugar.
(7) X in the general formula (1) is glucose, mannose, mannose-6-phosphate, fucose, galactose, glucosamine, lactosamine, galactosamine, N-acetylglucosamine, N-acetylgalactosamine, gluconic acid, glucuronic acid, galacturon. N according to any one of (1) to (6), which is an acid, lactose, maltose, isomaltose, sucrose, cellobiose, trehalose, lactobionic acid, apo B, transferrin, tuftsin, immunoglobulin, or folate. -Acylamine compounds.
(8) The N-acylamine compound according to any one of (1) to (7), which is biodegradable.
(9) Particles having at least a surface of at least one N-acylamine compound according to any one of (1) to (8).
(10) The particle according to (9), wherein the particle is a liposome.
(11) The particles according to claim 9, wherein the surface of the base particles is coated with at least one N-acylamine compound according to any one of (1) to (8).
(12) The particles according to any one of (9) to (11), wherein the base particles are made of a biodegradable material.
(13) The particles according to any one of (9) to (12), wherein the size of the particles is in the range of 0.1 nm to 5 mm.
(14) The particle according to any one of (9) to (13), wherein the particle contains at least one of a physiologically active substance, a drug, and a gene.
(15) The particle according to (14), wherein the physiologically active substance is at least one selected from growth factors, cytokines, hormones, and vasoactive substances.
(16) The drug is a steroid, a contrast agent, an anticancer drug, an anti-inflammatory drug, an antiallergic drug, a cardiovascular drug, an antihypertensive drug, a vasopressor drug, an antiplatelet drug, an anticoagulant drug, a psychotropic drug, a pain drug, an antibiotic. (14) The particle according to (14), which is at least one selected from a substance, an antiviral drug, a gastrointestinal drug, a urological drug, an endocrine drug, and a fluorescent reagent.
(17) The particle according to (14), wherein the gene is at least one selected from DNA and RNA.
(18) After dissolving the N-acylamine compound according to any one of (1) to (8) and a physiologically active substance or a drug in a water-soluble organic solvent, the solution is dropped into water, the organic solvent is removed, and centrifugation or A method for producing particles, wherein the coated particles are separated by any one of filtration.
(19) The N-acylamine compound according to any one of (1) to (8) and a physiologically active substance or a drug are dissolved in a water-insoluble organic solvent, dropped into water, the organic solvent is removed, and centrifugation is performed. Or a method for producing particles, wherein the coated particles are separated by either filtration or filtration.
(20) After dissolving the raw material of the particles and the physiologically active substance or drug in a water-insoluble organic solvent, the solution is dropped into an aqueous solution in which the N-acylamine compound according to any of (1) to (8) is dissolved, A method for producing coated particles, comprising removing an organic solvent and separating the coated particles by either centrifugation or filtration.
(21) After dissolving the N-acylamine compound according to any one of (1) to (8), the raw material of the particles, and the physiologically active substance or drug in a water-insoluble organic solvent, the solution is dropped into water, and the organic solvent is removed. A method for producing coated particles, comprising removing and separating the coated particles by either centrifugation or filtration.
(22) The method for producing particles according to (18) to (22), wherein the mixture is dropped into water while performing ultrasonic treatment.
(23) The method for producing particles according to (18) to (23), wherein the mixture is dropped into water while stirring.
(24) The method for producing particles according to (18) to (24), wherein the organic solvent is removed under reduced pressure.
(25) After dissolving the N-acylamine compound according to any one of (1) to (8) and a physiologically active substance or a drug in a water-soluble organic solvent, filling the dialysis tube and performing dialysis against water. A method for producing coated particles, liposomes, and micelles, comprising removing an organic solvent and separating the coated particles by either centrifugation or filtration.
(26) A medical device having at least a surface of the N-acylamine compound according to any one of (1) to (8).
(27) The medical device according to (26), wherein the medical composition is a catheter.
(28) The medical device according to (26), wherein the medical composition is a cell therapy system device.
(29) The medical device according to (26), wherein the medical composition is a device for a test / diagnosis drug.
(30) The medical device according to (26), wherein the medical composition is an implant device.
(31) The medical device according to (26), wherein the medical composition is an extracorporeal circulation system. "
[0030]
BEST MODE FOR CARRYING OUT THE INVENTION
Hereinafter, the present invention will be described in detail.
[0031]
The N-acylamine compound of the present invention has the following general formula:
[0032]
Embedded image

[0033]
(Where R ₁ Is an alkyl group having 1 to 23 carbon atoms or an alkenyl group having at least one double bond, R ₂ Is hydrogen or an alkyl or alkenyl group having 1 to 18 carbon atoms, R ₃ Is an alkylene group having 1 to 18 carbon atoms, A is any of O, S, COO, OCO, COS, SCO, CONR4, NR4CO, NHCOO, OCONH, NHCOS, SCONH, NHCONH (where R4 is hydrogen or carbon number 1 to 18 alkyl groups or alkenyl groups), and X is a biological substance).
[0034]
The N-acylamine compound of the present invention is characterized by comprising a substance derived from a living body. Accordingly, the structure of the constituent elements of the N-acylamine compound is basically not limited, as long as it is derived from a biological substance. R in the general formula (1) ₁ Forms a hydrophobic portion of the molecule, is preferably an alkyl or alkenyl group having 1 to 24 carbon atoms, and may be branched. Since it exists in a living body at a high ratio, it is most preferable that it has 11 to 19 carbon atoms, and more preferably it is linear. R ₁ Is an alkenyl group, it preferably contains 1 to 5 double bonds because it is present in a high proportion in a living body. R ₂ Is preferably hydrogen or an alkyl or alkenyl group having 1 to 18 carbon atoms. R ₁ And R ₃ -CONR which combines ₂ The bond is an amide bond, and at a temperature near body temperature, a hydrolysis reaction hardly occurs without an enzyme. In order for this bond to be hydrolytically cleaved by the enzyme, R ₂ Is most preferably hydrogen. Also, R ₂ Is a hydrogen atom, it is known that the amide bond forms a hydrogen bond with each other. Therefore, when this molecule is coated on the surface of the substrate, the molecules interact on the surface of the substrate to obtain a high surface density. R3 can be freely selected from C1 to C18, but is preferably an alkylene group having 2 to 4 carbon atoms because it is present in a high ratio in a living body. X is a substance characterized by being composed of a biological substance, and serves as a ligand that characterizes the surface to be coated. Specific examples of X include amino acids, oligopeptides, polypeptides, antibodies, parts of antibodies, receptors, enzymes, glucose, mannose, mannose-6-phosphate, galactose, glucosamine, lactosamine, galactosamine, N-acetylglucosamine Monosaccharides such as N-acetylgalactosamine, gluconic acid, glucuronic acid and galacturonic acid, lactose, maltose, isomaltose, sucrose, cellobiose, trehalose, oligosaccharides such as sialyl Lewis X, sialyl Lewis a, heparin, low molecular weight heparin, hyaluronic acid , Polysaccharides such as dermatan sulfate, chondroitin sulfate, keratan sulfate, and syndecane, but are not limited thereto.
[0035]
In particular, when this compound is used to impart affinity to the liver or hepatocytes, galactose, galactosamine, oligosaccharide or polysaccharide having galactose or galactosamine at the non-reducing end as X, and ApoB are preferably used. The sugar chain may be branched, and galactose or galactosamine is preferably present at each non-reducing end of the branched chain.
[0036]
In particular, when this compound is used to impart affinity to macrophages, X is preferably galactosamine, an oligosaccharide or polysaccharide having galactosamine at the non-reducing end, or Apo B is preferably used. The sugar chain may be branched, and galactosamine is preferably present at each non-reducing end of the branched chain. Tuftsin is also preferably used.
[0037]
In particular, when this compound is used for imparting affinity to T cells, X is preferably gp120 g derived from the HIV virus.
[0038]
In particular, when this compound is used for imparting affinity to cancer cells, X is preferably transferrin, EGF, tuftsin, or folic acid.
[0039]
R in general formula 1 ₁ ~ R ₃ The solubility of the N-acylamine compound in the solvent can be freely changed by selecting the structures of X and X.
[0040]
Biological substances X and R ₃ Can be freely selected according to the chemical structure, orientation, and degradability of X, and are easily hydrolyzed, so that ester bonds, ureido bonds, and enzymatic degradation that are susceptible to hydrolysis are obtained. Amide bond, glucoside bond, ether bond or thioether bond typified by galactoside bond typified by a peptide bond which is susceptible to glycerol is preferable, that is, specific examples of A include O, S, COO, OCO, COS, SCO, CONR4, Any of NR4CO, NHCOO, OCONH, NHCOS, SCONH, and NHCONH (here, R4 is hydrogen or an alkyl group or alkenyl group having 1 to 18 carbon atoms).
[0041]
The N-acylamine compound of the present invention can be produced by any method. As shown in the general formula (1), -CONR ₂ Since the amide bond of-is essential, the N-acylamine compound of the present invention comprises a carboxylic acid containing R1 and a carboxylic acid containing R1. ₂ And R ₃ It is most preferable to use an amino compound containing the compound and a biological substance X as starting materials from the viewpoint of ease of synthesis. It is preferable that the amino compound has another reactive functional group other than the amino group from the viewpoint of binding the biological substance X. Specific examples of the reactive functional group include an amino group, a hydroxyl group, a carboxyl group, a thiol group, an aldehyde group, an isocyanate group, and a glycidyl group.
[0042]
There is no particular limitation on the order in which the respective starting materials are combined, and the biological material X may be reacted with a carboxylic acid after being bonded to the amino compound by the bond represented by A. X may be bonded after bonding to obtain an intermediate amide compound. Amide bond-CONR ₂ The intermediate amide compound containing- ₁ A carboxylic acid or a derivative thereof containing ₂ And R ₃ By reacting with an amino compound containing Derivatives refer to carboxylic acid chlorides, carboxylic acid bromides, carboxylic acid iodides, carboxylic esters, carboxylic anhydrides, and carboxylic acid salts. The intermediate amide compound is obtained by activating a carboxylic acid with a carbodiimide such as dicyclohexylcarbodiimide, adding an amino compound, removing a urea compound such as dicyclohexylurea generated as a by-product, and then removing the solvent, and then obtaining the intermediate amide compound with a carbodiimide. After the activation, the carboxyl group is further activated with N-hydroxysuccinimide, and then an amino compound is added. In addition, an amino compound is added to carboxylic acid chloride, carboxylic acid bromide, carboxylic acid iodide, or carboxylic anhydride in the presence of a catalyst such as pyridine, triethylamine, or dimethylaminopyridine, and the solvent is removed after removing the salt of the catalyst. It is also obtained by doing. Furthermore, it can be obtained by adding an amino compound to a carboxylic acid ester and removing the solvent. A reactive functional group other than the amino group of the amino compound is protected by a protecting group such as a tert-butoxycarbonyl group, a carbobenzoxy group, a nitrophenyl group, a formyl group, an acyl group, a haloacyl group, a benzoyl group, and a tosyl group. In such a case, it is preferable to remove the protecting group after the amide bond reaction is completed.
[0043]
The method for bonding the biological compound X with the amino compound or the intermediate amide compound is not particularly limited and can be appropriately selected.
[0044]
When a sugar represented by monosaccharide, oligosaccharide or polysaccharide is bonded by an ether bond or a thioether bond as X, generally, the hydroxyl group of the sugar is first protected by an acyl group, a haloacyl group, a benzylidene group, an iso-propylidene group, or the like. I do. The protection reaction is described in detail in Hatanaka et al., "Science and Engineering of Carbohydrates" (Kodansha). An acetyl group or a haloacetyl group is preferably used because of easy protection and deprotection. A protected sugar is dissolved in a solvent such as chloroform or dichloroethane, and an intermediate amide compound having a hydroxyl group or a thiol group or a starting material having an amino group protected in the presence of a Lewis acid catalyst such as tin tetrachloride or boron trifluoride. After reacting with the addition of the amino compound, the product obtained by removing the catalyst and the solvent is treated with a Lewis base such as sodium methoxide to remove the protecting group to obtain a sugar-modified N-acylamine compound. . In the sugar-modified N-acylamine compound obtained by this reaction, the sugar is bonded at the anomeric carbon at the reducing end. The same applies to the case where the carbon at the anomeric position is brominated with hydrogen bromide instead of the Lewis acid catalyst, and then reacted with an intermediate amide compound or an amino group-protected starting material amino compound in the presence of silver perchlorate. A sugar-modified N-acylamine compound is obtained.
[0045]
As a general method of linking a sugar with an amide bond, an intermediate amide compound having a carboxyl group or a starting amino compound having an amino group protected is activated with a carbodiimide such as dicyclohexylcarbodiimide to form an amino group such as galactosamine or chitosan. Is obtained by removing the solvent after removing a urea compound such as dicyclohexylurea which is produced as a by-product, and after activating with carbodiimide, further activating the carboxyl group with N-hydroxysuccinimide. Can also be obtained by adding sugar. If the sugar contains a carboxyl group such as gluconic acid or glucuronic acid, after protecting the hydroxyl group, the sugar is activated with carbodiimide to give an intermediate amide compound having an amino group or an amino group-protected starting compound. A raw material amino compound is added and obtained by removing a solvent after removing a urea compound such as dicyclohexylurea generated as a by-product, and after activating with carbodiimide, further activating a carboxyl group with N-hydroxysuccinimide. Can also be obtained by adding sugar.
[0046]
As a general method of linking a sugar with an ester bond or a thioester bond, when a sugar contains a carboxyl group such as gluconic acid or glucuronic acid, the hydroxyl group is protected with a benzyl group or the like, and then the sugar is converted into a salt. After treatment with thionyl, phosphorus trichloride, phosphorus pentachloride and the like to chlorinate the carboxyl group, an intermediate amide compound containing a hydroxyl group or a thiol group or an amino group is added in the presence of a catalyst such as pyridine, triethylamine and dimethylaminopyridine. It is obtained by adding the protected starting amino compound and removing the salt and solvent of the catalyst.
[0047]
An intermediate amide compound containing a carboxyl group or a compound in which the carboxyl group of the starting amino compound protected with an amino group is previously converted to an acid chloride, acid bromide, acid iodide, or acid anhydride, is converted to pyridine, triethylamine, dimethylamino. It can also be obtained by adding to a sugar in the presence of a catalyst such as pyridine and removing the salt and solvent of the catalyst.
[0048]
When X is an amino acid, an oligopeptide, a polypeptide, a protein, or a peptide compound typified by a fragment of a protein, which is bonded by an amide bond, an intermediate amide compound having a carboxyl group or a starting material protected with an amino group is generally used. An amino compound is activated with carbodiimide, an amino acid or a polypeptide containing an amino group is added, the urea compound such as dicyclohexylurea generated as a by-product is removed, and then the solvent is removed and then the carbodiimide is obtained. After that, the carboxyl group is further activated with N-hydroxysuccinimide, and then the peptide compound is added. According to this method, the peptide compound can be bound with the amino group at the amino terminus or the amino group of a lysine residue. When a peptide compound is bonded by an ester bond or a thioester bond, generally, an intermediate amide compound having a carboxyl group or a starting amino compound protected with an amino group is converted to a thionyl chloride, phosphorus trichloride, phosphorus pentachloride, or the like. And then chlorinating the carboxyl group, adding an amino compound containing a hydroxyl group or a thiol group in the presence of a catalyst such as pyridine, triethylamine, or dimethylaminopyridine, and removing the salt and solvent of the catalyst. . By using this method, the peptide compound can be bound by a hydroxyl group of a serine residue or a threonine residue and a thiol group of a cysteine residue. Further, a peptide compound having a carboxyl group is treated with thionyl chloride, phosphorus trichloride, phosphorus pentachloride, etc. to chlorinate the carboxyl group, and then, in the presence of a catalyst such as pyridine, triethylamine, dimethylaminopyridine, a hydroxyl group or a thiol group. Can also be obtained by adding an intermediate amide compound or a starting amino compound in which the amino group is protected, and removing the salt of the catalyst and the solvent. According to this method, the peptide compound can be bonded with a carboxyl group at the carboxy terminus, an aspartic acid residue, or a glutamic acid residue.
At this time, when the peptide compound contains an amino group, it is preferably protected with a protecting group, and the protecting group can be removed after the completion of the binding reaction.
[0049]
The solvent, reaction temperature, reaction time, and purification method can be appropriately selected depending on the chemical structure of the raw materials and precursor substances and the type of reaction.
[0050]
The term "biodegradable" as used in the present invention means a property that is degraded in a living body, and is mainly decomposed by a hydrolysis reaction, regardless of whether an enzyme is involved or not, to produce a biocompatible substance. . A biocompatible substance is a substance that has the same chemical structure as a substance that originally exists in a living body, or that can be metabolized and excreted by enzymes, immunity, liver function, kidney function, and the like. If a plurality of constituents formed by decomposing the N-acylamine compound of the present invention are substances derived from a living body, they are biocompatible substances, that is, the N-acylamine compound is biodegradable.
[0051]
The N-acylamine compound of the present invention is preferably used as a surface-modifying molecule processed into a form present at least on the surface. For processing into the form present on the surface, the N-acylamine compound of the present invention may be used by coating it on the surface of the base particles, and the liposome or micelle, that is, the N-acylamine compound itself is used to form particles. You may. In this case, the type of the N-acylamine compound to be used can be appropriately adjusted depending on the purpose and is not limited. One type of N-acylamine compound may be used alone, or a plurality of N-acylamine compounds may be used as a mixture. The liposome here means that the N-acylamine compound has R ₁ And a double film is formed by associating with each other to form a double film. Further, a micelle is a compound in which an N-acylamine compound is ₁ Is a spherical composition in which a core is formed by associating with each other, and X is distributed on the surface.
[0052]
In this case, the shape of the liposome, micelle, or particle is not particularly limited. The size is not particularly limited, and a size suitable for the purpose can be selected. The size of the cell gap varies from tissue to tissue, and the size that cells can take up also varies from cell to cell. If the size is too large, it cannot be taken up into tissues and cells, while if the size is too small, it will be easily taken up by tissues and cells other than the target. Further, for the purpose of adsorbing particles on the surface of a tissue or a cell, a size that is not taken up by the tissue or the cell is preferable. The average particle size is preferably from 0.1 nm to 5 mm. For the purpose of incorporation into tissues or cells, the thickness is preferably 0.1 nm to 500 μm, and particularly for the purpose of incorporation into liver or hepatic parenchymal cells, more preferably 10 nm to 500 nm. The average particle diameter of the particles is obtained by measuring the particle diameters of a plurality of particles using a scanning electron microscope S-2400 (manufactured by Hitachi, Ltd.) and calculating the average value. When a laser scattering particle size distribution analyzer (Microtrac ASVR / Microtrac-HRA (9320-X100)) is used, the average particle size can be directly known.
[0053]
When the N-acylamine compound of the present invention is used after being coated on the surface of a base particle, the base material constituting the particle is not particularly limited, but is decomposed, metabolized, and excreted by enzymes, immunity, liver function, kidney function and the like. Preferably, it consists of a substance. Specific examples of such substances include polyethylene glycol, polylactic acid, polyglycolic acid, polycaprolactone, polyvinyl alcohol, cellulose, agarose, starch, alginic acid, chitin, chitosan, and derivatives and copolymers of these substances. But not limited thereto. When used for the purpose of being taken up by cells, it is preferably biodegradable, and polylactic acid, polyglycolic acid, polycaprolactone and copolymers thereof are preferably used. A known method can be used for measuring the biodegradability of the particle substrate. For example, immersing the material in water or salt solution at an appropriate solid-liquid ratio and shaking or stirring at an appropriate temperature for an appropriate period, A method of immersing in a buffered saline and shaking or stirring at 37 ° C. is preferably used. The biodegradability is evaluated by measuring the dry weight, volume, average molecular weight, appearance, etc. of the material using a balance, gel permeation chromatography, scanning electron microscope, etc., and examining the change over time.
[0054]
If the density of the N-acylamine compound of the present invention on the particle surface is too high, not only the orientation of the ligand is impaired, but also steric hindrance occurs, making it difficult for the receptor present on the cell to be recognized. On the other hand, if it is too low, the chance of contact between the ligand and the receptor is reduced, so that it is difficult for the cell to recognize the receptor. Therefore, the surface density of the particle surface is 1 to 1000 nm ² Is preferably one molecule.
[0055]
When the N-acylamine compound of the present invention is used as a liposome or coated on the surface of a base particle, the liposome or the base particle contains a single or a plurality of physiologically active substances, drugs, and genes. Is also good. Examples of the physiologically active substance include growth factors such as PDGF, VEGF, HGF, FGF, and EGF, cytokines such as INF, TGF, and interleukin, and hormones. Also included are antigens such as killed bacteria, toxins, sugar chains, and peptides used as vaccines. Drugs include contrast agents, anticancer drugs, anti-inflammatory drugs, antiallergic drugs, cardiovascular drugs, antihypertensive drugs, vasopressors, antiplatelet drugs, anticoagulants, psychotropic drugs, pain drugs, antibiotics, and digestive organs Drugs, urological drugs, endocrine drugs, fluorescent reagents and the like. Examples of the gene include an oligonucleotide, a polynucleotide, DNA, and RNA. In particular, when targeting the liver or hepatocytes, antiviral agents such as ribavirin, lamivucine and interferon, anti-inflammatory agents such as cyclosporin, anticancer agents such as cisplatin, carboplatin, 5-Fu, mitomycin, cyclophosphamide, and methotrexate. X-ray contrast agents such as, Iopamidol, Iohexol, ¹³¹ I, ^99m Tc, ¹³¹ I-HSA, ⁶⁷ Ga, ³ H, ²⁴ Na, ⁸⁶ Rb, ^87m Sr, ¹⁸ Contrast agents for scintigraphy such as F-FDG, Gd compounds, Fe compounds, contrast agents for nuclear magnetic resonance such as Mn compounds, polydextrose, ultrasound contrast agents such as octafluoropropane, hyperlipidemia agents such as fibrate, Growth factors such as HGF and angiogenesis inhibitors such as endostatin and angiostatin are preferably used.
[0056]
The method for preparing the particles coated with the N-acylamine compound in the present invention is not particularly limited, but well-known spray methods and emulsion methods are preferably used. The water-soluble solvent referred to in the present invention is a solvent that is easily dissolved in water such as methanol, ethanol, acetone, DMSO, DMF, and DMAc. The water-insoluble solvent is a water-soluble solvent such as chloroform, dichloromethane, diethyl ether, and ethyl acetate. Refers to a solvent that is insoluble in
[0057]
In the emulsion method, a particle base material is dissolved in a water-insoluble solvent such as chloroform or methylene chloride, dropped into water in which the N-acylamine compound of the present invention is dissolved, and stirred to evaporate the organic solvent. By appropriately selecting the concentration of the N-acylamine compound of the present invention, the concentration of the particle base material, the number of rotations for stirring, the temperature and the pressure, the size of the particles can be appropriately changed. After stirring, the coated particles are washed with distilled water, and the coated particles are separated from water by centrifugation or filtration to obtain coated particles. If the particle base concentration is too high, not only is the operability of the solution poor, but also the base particle solution does not disperse in the aqueous solution of the N-acylamine compound, and if the concentration is too low, the obtained particles are small and the efficiency is low. 001 to 30 w / v% is preferable, and 0.01 to 20 w / v% is particularly preferable. If the stirring speed is too high, the particle size varies, and if the stirring speed is too low, fine particles cannot be obtained. Therefore, the stirring speed is preferably 100 to 1000 rpm, and more preferably 100 to 400 rpm. The temperature condition depends on the solvent that dissolves the particle base material.If it is too high, the solvent will evaporate immediately, making it difficult to obtain fine particles, and the dispersibility of the emulsion will be poor.If it is too low, the solvent will not evaporate and the particles will not evaporate. Since the preparation takes a long time, the temperature is preferably from 0 ° C. to the boiling point of the solvent. In order to promote the evaporation of the solvent, the operation can be performed under reduced pressure, and the pressure at this time is preferably 1 atm or less. For example, the particle substrate is dissolved in a water-insoluble solvent such as chloroform or methylene chloride so as to have a concentration of 0.01 to 20% w / v. -Dropping the acylamine compound in water having a concentration of 0.01 to 10% in water, stirring at a suitable temperature at a speed of 100 to 400 revolutions / minute, and separating the coated particles from the water by centrifugation or filtration. The surface density of the N-acylamine compound is 1 to 1000 nm ² Particles coated with an N-acylamine compound having an average particle size of 0.1 nm to 5 mm per molecule are obtained.
[0058]
Further, the N-acylamine compound of the present invention and the particle base material are dissolved in a water-insoluble solvent such as chloroform and methylene chloride so that the concentrations thereof are 0.01 to 10% and 0.001 to 10%, respectively. Is dropped into water having a volume of 10 to 1000 times the volume of the particle base solution, and the mixture is stirred at a suitable temperature, preferably at a rate of 100 to 1000 rpm, more preferably at a rate of 100 to 400 rpm, and coated by centrifugation or filtration. Coated particles are also obtained by separating the particles from water.
[0059]
Stirring may be performed while treating with ultrasonic waves, or only ultrasonic treatment may be performed without stirring. For example, when using a US-50 probe type ultrasonic apparatus (Nissei), if the output is too low, the particle base material solution will not disperse in water, and if the output is too high, the average particle diameter will be small. Is preferred.
[0060]
After dissolving the N-acylamine compound of the present invention and the particle base material in a water-soluble solvent such as DMSO, DMF, methanol, ethanol, or acetone, preferably at a concentration of 0.001 to 30 w / v%, the solution is dialyzed. After dialysis in water in a tube, the surface density of the N-acylamine compound is 1 to 1000 nm by separating the coated particles from the water by centrifugation or filtration of the suspension in the dialysis tube. ² / Coated particles having an average particle size of 0.1 nm to 5 mm per molecule are obtained.
[0061]
When preparing liposomes or micelles, the N-acylamine compound of the present invention may be dissolved directly in water, preferably at a concentration of 0.001 to 30 w / v%, or may be DMSO, DMF, methanol, ethanol, acetone. After dissolving in a water-soluble solvent at a concentration of preferably 0.001 to 30 w / v%, the solution may be dropped into water having a volume of 1 to 1000 times the volume of the particle base solution. Preferably, liposomes or micelles having a uniform particle size are obtained by ultrasonic treatment after dripping into water. Liposomes or micelles are obtained by separating coated particles from water by centrifugation or filtration.
[0062]
When a physiologically active substance or drug is to be carried on particles, liposomes or micelles, the above operation is performed after the physiologically active substance or drug or their solution is allowed to coexist in the solution of the particle base material. The concentration of the physiologically active substance or drug in the particles can be appropriately selected from the charged concentration and the like, and can be changed according to the intended concentration. For example, in the emulsion method, when a physiologically active substance or a drug is hydrophobic, the particle base and the drug are dissolved and mixed in a water-insoluble solvent such as chloroform or methylene chloride, and the N-acylamine compound of the present invention is dissolved in 0.1%. It is obtained by dripping into water dissolved to a concentration of 01 to 10% and stirring at an appropriate temperature at a rate of 100 to 400 revolutions / minute. When the physiologically active substance or drug is hydrophilic, an emulsion is prepared by stirring and mixing an aqueous solution of the drug and a solution in which the particle base material is dissolved in a water-insoluble solvent such as chloroform or methylene chloride. Is dropped into water in which the N-acylamine compound of the present invention is dissolved to a concentration of 0.01 to 10%, and the mixture is stirred at a suitable temperature at a rate of 100 to 400 revolutions / minute. The concentration of the physiologically active substance or drug can be freely adjusted to the concentration necessary for exhibiting the desired drug effect. When an emulsion is prepared, a dispersant such as polyethylene glycol or polyvinyl alcohol may be used. Depending on the structure, the N-acylamine compound itself may serve as a dispersant, so that the N-acylamine compound may be used instead of a dispersant such as polyethylene glycol or together with the dispersant when preparing the emulsion. When a physiologically active substance or a drug is contained, the size of the particles can be appropriately changed by appropriately selecting the concentration of the N-acylamine compound, the concentration of the particle base material, the number of rotations of stirring, and the temperature. After stirring and washing with distilled water, the particles are obtained by separating the coated particles from the water by centrifugation or filtration. If the particle base concentration is too high, not only is the operability of the solution poor, but also the base particle solution does not disperse in the aqueous solution of the N-acylamine compound, and if the concentration is too low, the obtained particles are small and the efficiency is low. 001 to 30 w / v% is preferable, and 0.01 to 20 w / v% is particularly preferable. If the stirring speed is too high, the particle size varies, and if the stirring speed is too low, fine particles cannot be obtained. Therefore, the stirring speed is preferably 100 to 1000 rpm, and more preferably 100 to 400 rpm. The temperature condition depends on the solvent that dissolves the particle base material.If it is too high, the solvent will evaporate immediately, making it difficult to obtain fine particles, and the dispersibility of the emulsion will be poor.If it is too low, the solvent will not evaporate and the particles will not evaporate. Since the preparation takes a long time, the temperature is preferably from 0 ° C. to the boiling point of the solvent.
[0063]
When preparing a liposome or micelle containing a physiologically active substance or a drug, the N-acylamine compound of the present invention is preferably dissolved in an aqueous solution such as DMSO, DMF, methanol, ethanol or acetone at a concentration of 0.001 to 30 w / v%. After dissolving in a basic solvent, adding a drug and mixing, the solution may be dropped into water having a volume of 1 to 1000 times the volume of the particle base solution. Preferably, liposomes or micelles having a uniform particle size are obtained by ultrasonic treatment after dripping into water.
[0064]
The medical device in the present invention is a device used for diagnosing, treating, and preventing a disease, and specifically, a carrier or vector containing a physiologically active substance, a drug, a gene, a drug delivery system such as a vaccine, and a catheter. Types, implant devices such as artificial blood vessels and artificial heads, blood circuits such as cardiopulmonary bypass, artificial dialysis machines, base materials for extracorporeal circulation systems such as adsorbents, base materials for test and diagnostic agents such as microplates and particles, cells It refers to, but is not limited to, a substrate for a cell therapy / regenerative medicine system, such as a scaffold or a cell carrier, and a chromatography carrier.
[0065]
By disposing an N-acylamine compound on the surface of these medical devices, biocompatibility is significantly improved. For example, catheters and blood circuits activate platelets and the coagulation cascade to form thrombi when they come into contact with blood.For example, this is achieved by coating the surface with an N-acylamine compound bound to heparin, an antithrombotic substance. Thrombus formation reaction on the surface of the base material can be suppressed. In addition, in the case of an implant device or a substrate for cell therapy / regenerative medicine system, the surface of the substrate induces a foreign substance reaction or an inflammatory reaction. By coating the surface with the acylamine compound, the affinity and the adhesion to tissues and cells at the target site are improved. Also, in laboratory diagnostics and extracorporeal circulation treatment, N-acylamine compounds, which bind substances interacting with substances to be quantified and substances to be removed outside the body, are coated on particles or fibers to quantify and remove those substances. it can.
[0066]
The method of applying the N-acylamine compound to the surface of the medical device is not particularly limited, and the medical device can be obtained by immersing the medical device in a solution of the N-acylamine compound, followed by drying to remove the solvent. Alternatively, molding may be performed after mixing the raw material of the medical device and the N-acylamine compound.
[0067]
Examples are shown below, but the present invention is not limited by these examples.
[0068]
【Example】
<Synthesis example 1>
After suspending 20 g of D-lactose in 320 ml of pyridine, 320 ml of acetic anhydride was added dropwise while stirring in an ice bath. After stirring at room temperature for 3 days, the reaction solution was poured into 700 ml of water. After the precipitate was separated by filtration, the precipitate was washed with water until the pH became 7. After drying under reduced pressure, 26 g of the product was dissolved in a mixed solvent of 70 ml of diethyl ether and 20 ml of ethyl acetate, sodium sulfate was added, and the mixture was stirred. The sodium sulfate was filtered off, and the solvent was removed under reduced pressure to obtain 27 g of acetylated lactose. .
[0069]
After dissolving 10 g of linoleic acid and 4.9 g of N-hydroxysuccinimide in dichloromethane, 8.8 g of dicyclohexylcarbodiimide and a catalytic amount of pyridine were added, and the mixture was stirred at room temperature for 3 days. After dicyclohexylurea was filtered off, the solvent was removed under reduced pressure to obtain 17 g of activated linoleic acid. After dissolving the whole amount in dichloromethane again, 3 ml of 2-aminoethanol was added, and the mixture was stirred at room temperature for 2 days. After the precipitate was separated by filtration, the solvent was removed under reduced pressure, and the obtained product was washed with water and dried to obtain linoleic acid amide.
[0070]
After dissolving 2 g of acetylated lactose and 3.8 g of linoleic acid amide in 70 ml of dichloroethane, 2 g of synthetic zeolite powder was added, and the mixture was stirred at 70 ° C. in a nitrogen gas atmosphere. After adding 1 ml of a boron trifluoride solution in diethyl ether, the mixture was stirred for 5 hours under the same conditions. The solid phase was filtered off with Hyflo Supercell, and the solvent was removed under reduced pressure. After dissolving in ether, the solution was administered in 2-propanol and reprecipitated. The precipitate was separated by filtration and dried under reduced pressure to obtain an acetylated lactose-linoleic acid amide complex.
[0071]
After dissolving 2 g of the acetylated lactose-linoleic acid amide complex in 200 ml of methanol, 600 mg of sodium methoxide was added, and the mixture was stirred at room temperature for 6 hours. After adding 35 g of a cation exchange resin (Amberlite IR-120B) and stirring, the resin was separated by filtration and the solvent was removed under reduced pressure to obtain 1.6 g of a lactose-linoleic acid amide complex.
[0072]
<Comparative Example 1>
A polylactic acid solution obtained by dissolving 0.6 g of polylactic acid (Wako Pure Chemical; average molecular weight: 5000) in 6 ml of dichloromethane was dropped into 400 ml of an aqueous solution in which 0.33 g of polyvinyl alcohol (PVA) (Nacalai Tesque; degree of polymerization: 500) was dissolved. The mixture was stirred at room temperature for 8 hours to obtain polylactic acid particles.
[0073]
A 1 mg / ml solution of PVLA (Seikagaku Corporation; average molecular weight: 50,000) in PBS or PBS was added to the polylactic acid particles, and the mixture was allowed to stand at room temperature for 1 hour and 50 minutes. After washing the particles with 0.5 ml of PBS, 0.2 ml of a PBS solution of 0.1 mg / ml FITC-labeled RCA120 (Honen Corporation) was added, and the mixture was allowed to stand at room temperature for 1 hour. After washing the particles with 0.5 ml of PBS, fluorescence on the surface of the particles was observed with a scanning laser biological microscope (FLOVIEW; Olympus). The setting conditions of the laser microscope were as follows.
[0074]

As a result, fluorescence was observed on the surface of the poly- (Np-vinyl benzyl-O-β-D-galactopyranosyl- [1 → 4] -D-glucoconamide (PVLA) -modified particles, but the particles were modified with PVLA. No fluorescence was observed on the surface of the missing particles (FIG. 1).
[0075]
<Example 1>
A lactose-linoleic acid amide complex PBS solution was added to the polylactic acid particles, and the mixture was allowed to stand at room temperature for 1 hour and 50 minutes. After washing the particles with 0.5 ml of PBS, 0.2 ml of a PBS solution of 0.1 mg / ml FITC-labeled RCA120 (Honen Corporation) was added, and the mixture was allowed to stand at room temperature for 1 hour. After washing the particles with 0.5 ml of PBS, the fluorescence of the particle surface was observed under the setting conditions described in Comparative Example 1 using a scanning laser biological microscope (FLVIEW, Olympus). Comparable fluorescence was observed (FIG. 1).
[0076]
<Comparative Example 2>
A polylactic acid solution obtained by dissolving 10 mg of polylactic acid (Wako Pure Chemical; molecular weight: 20,000) in 0.1 ml of dichloromethane is added to 0.5 ml of an aqueous solution obtained by dissolving 0.5 mg of PVLA (Seikagaku Corporation) or PVA (Nacalai Tesque; degree of polymerization 500). Was dropped with a Pasteur pipette and stirred at room temperature for 1 day to obtain lactose-coated polylactic acid particles.
[0077]
After the polylactic acid particles were washed with 0.5 ml of PBS, 0.2 ml of a 0.1 mg / ml FITC-labeled RCA120 (Honen Corporation) in 0.2 ml of PBS was added, and the mixture was allowed to stand at room temperature for 1 hour. After washing the particles with 0.5 ml of PBS, fluorescence was observed on the surface of the particles with a scanning laser biological microscope (FLOVIEW; Olympus) under the setting conditions described in Comparative Example 1, and as a result, fluorescence was observed (FIG. 2). The average fluorescence intensity on any three planes in the fluorescent coloring portion was 4073 ± 21 for particles coated with PVLA and 157 ± 4 for particles coated with PVA.
[0078]
<Example 2>
A polylactic acid solution obtained by dissolving 10 mg of polylactic acid (Wako Pure Chemical Industries, Ltd., molecular weight: 20,000) in 0.1 ml of dichloromethane was dropped with a Pasteur pipette into a few drops of an aqueous solution in which 1 mg of lactose-linoleic acid amide complex was dissolved. For 1 day to obtain lactose-coated polylactic acid particles.
[0079]
After the polylactic acid particles were washed with 0.5 ml of PBS, 0.2 ml of a 0.1 mg / ml FITC-labeled RCA120 (Honen Corporation) in 0.2 ml of PBS was added, and the mixture was allowed to stand at room temperature for 1 hour. After washing the particles with 0.5 ml of PBS, the fluorescence of the particle surface was observed with a scanning laser biological microscope (FLOVIEW; Olympus) under the setting conditions described in Comparative Example 1. As a result, the PVLA-modified particles of Comparative Example 2 were observed on the particle surface. Comparable fluorescence was observed (FIG. 2). The average fluorescence intensity on any three planes in the fluorescent coloring portion was 3941 ± 56.
[0080]
<Comparative Example 3>
The cells were treated with a homogenizer at a low temperature, suspended in phosphate buffered saline, and then ultracentrifuged to remove insolubles, thereby preparing a cell extract.
[0081]
5 mg of PVLA was dissolved in 0.1 ml of methanol, 20 μl of 10 μg / ml fluorescein was added, and then this solution was added in 0.9 ml of phosphate buffered saline. The cell extract was added and shaken at 37 ° C. for 24 hours. The reaction solution was placed in a dialysis membrane and dialyzed against distilled water. After freeze-drying the liquid in the tube, molecular weight analysis was performed by high performance liquid chromatography. As a result, the molecular weight of PVLA before and after treatment with the cell extract was about 50,000, which was unchanged and was not decomposed by intracellular enzymes.
[0082]
<Example 3>
The cells were treated with a homogenizer at a low temperature, suspended in phosphate buffered saline, and then ultracentrifuged to remove insolubles, thereby preparing a cell extract.
[0083]
5 mg of lactose-linoleic acid amide complex was dissolved in 0.1 ml of methanol, 20 μl of 10 μg / ml fluorescein was added, and then this solution was added to 0.9 ml of phosphate buffered saline. The cell extract was added, and the time-dependent change in the fluorescence intensity at 37 ° C. was measured using a fluorescence spectrophotometer (F-2000; Hitachi, Ltd.) under the following conditions.
[0084]
Excitation wavelength: 490 nm
Scan speed: 60 nm / min
Response: 2 seconds
Band pass: excitation side 10 nm, emission side 10 nm
Photomultiplier tube: 400V
The fluorescence intensity changes over time from 1608 → 6 → 30, reaching the same level as when fluorescein was added to linoleic acid amide and linoleic acid, proving that lactose-linoleic acid amide complex is degraded by intracellular enzymes. Was done.
[0085]
【The invention's effect】
Since the surface coating compound of the present invention is decomposed in the body, a drug or the like can be safely delivered to a target tissue or cell by coating with the present compound.
[Brief description of the drawings]
FIG. 1 is a laser micrograph of particles coated with PVLA and lactose-linoleic acid complex of Example.
FIG. 2 is a laser micrograph of particles coated with PVLA and lactose-linoleic acid complex of Example.
FIG. 3 is a change in fluorescence when fluorescein and a cell extract are added to the lactose-linoleic acid complex of Example.

Claims (31)

The following general formula (1)

(Where R ₁ is an alkyl group having 1 to 23 carbon atoms or an alkenyl group having at least one double bond, R ₂ is hydrogen or an alkyl group or alkenyl group having 1 to 18 carbon atoms, and R ₃ is a C 1 to C 18 group. Wherein A is any of O, S, COO, OCO, COS, SCO, CONR4, NR4CO, NHCOO, OCONH, NHCOS, SCONH, NHCONH (where R4 is hydrogen or an alkyl group having 1 to 18 carbon atoms) Or an alkenyl group) and X is an N-acylamine compound represented by a biological substance). N- acyl amine compound according to claim 1 wherein R ₂ in the general formula (1) is characterized in that it is a hydrogen. The N-acylamine compound according to claim 1, wherein the alkyl group or alkenyl group of R _{1 in} the general formula (1) is linear having 12 to 23 carbon atoms. R ₁ is an alkenyl group in the general formula (1), and N- acyl amine compound according to claim 1, wherein the number of double bonds is 1-5. The N-acylamine compound according to claim 4, wherein R3 in the general formula (1) is an alkylene group having 1 to 4 carbon atoms. The N-acylamine compound according to any one of claims 1 to 5, wherein X in the general formula (1) is selected from sugar. X in the general formula (1) is glucose, mannose, mannose-6-phosphate, fucose, galactose, glucosamine, lactosamine, galactosamine, N-acetylglucosamine, N-acetylgalactosamine, gluconic acid, glucuronic acid, galacturonic acid, lactose The N-acylamine compound according to any one of claims 1 to 6, wherein the compound is maltose, isomaltose, sucrose, cellobiose, trehalose, lactobionic acid, apo B, transferrin, tuftsin, immunoglobulin, or folic acid. The N-acylamine compound according to any one of claims 1 to 7, which is biodegradable. A particle having at least one N-acylamine compound according to any one of claims 1 to 8 on a surface thereof. The particle according to claim 9, wherein the particle is a liposome. The particles according to claim 9, wherein the surface of the base particles is coated with at least one N-acylamine compound according to any one of claims 1 to 8. The particles according to any one of claims 9 to 11, wherein the base particles are made of a biodegradable material. 13. Particles according to any of claims 9 to 12, characterized in that the size of the particles is in the range from 0.1 nm to 5 mm. The particle according to any one of claims 9 to 13, wherein the particle contains at least one of a physiologically active substance, a drug, and a gene. The particle according to claim 14, wherein the physiologically active substance is at least one selected from a growth factor, a cytokine, a hormone, and a vasoactive substance. The drug is a steroid, a contrast agent, an anticancer drug, an anti-inflammatory drug, an antiallergic drug, a cardiovascular drug, an antihypertensive drug, a vasopressor drug, an antiplatelet drug, an anticoagulant, a psychotropic drug, a pain treatment drug, an antibiotic, The particle according to claim 14, wherein the particle is at least one selected from a viral drug, a digestive drug, a urological drug, an endocrine drug, and a fluorescent reagent. The particle according to claim 14, wherein the gene is at least one selected from DNA and RNA. After dissolving the N-acylamine compound according to any one of claims 1 to 8 and a physiologically active substance or a drug in a water-soluble organic solvent, the solution is dropped into water, the organic solvent is removed, and centrifugation or filtration is performed. A method for producing particles, comprising separating coated particles. The N-acylamine compound according to any one of claims 1 to 8 and a physiologically active substance or a drug are dissolved in a water-insoluble organic solvent, dropped into water, the organic solvent is removed, and any of centrifugation or filtration is performed. A method for producing particles, wherein the coated particles are separated by a method. After dissolving the raw material of the particles and the physiologically active substance or drug in a water-insoluble organic solvent, the solution is dropped into an aqueous solution in which the N-acylamine compound according to any one of claims 1 to 8 is dissolved to remove the organic solvent. A method for producing coated particles, wherein the coated particles are separated by any of centrifugation and filtration. After dissolving the N-acylamine compound according to any one of claims 1 to 8 and the raw material of the particles and the physiologically active substance or drug in a water-insoluble organic solvent, the solution is dropped into water to remove the organic solvent, and centrifuged. A method for producing coated particles, wherein the coated particles are separated by either filtration or filtration. The method for producing particles according to claim 18, wherein the particles are dropped into water while performing ultrasonic treatment. The method for producing particles according to claim 18, wherein the mixture is dropped into water while stirring. The method for producing particles according to claim 18, wherein the organic solvent is removed under reduced pressure. After dissolving the N-acylamine compound according to any one of claims 1 to 8 and a physiologically active substance or a drug in a water-soluble organic solvent, filling the dialysis tube and performing dialysis against water to remove the organic solvent. A method for producing coated particles, liposomes, and micelles, wherein the coated particles are separated by any of centrifugation and filtration. A medical device having at least the surface of the N-acylamine compound according to claim 1. The medical device according to claim 26, wherein the medical composition is a catheter. 27. The medical device of claim 26, wherein the medical composition is a cell therapy system device. The medical device according to claim 26, wherein the medical composition is a device for a test / diagnosis drug. The medical device according to claim 26, wherein the medical composition is an implant device. The medical device according to claim 26, wherein the medical composition is an extracorporeal circulation system.

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