JP4830181B2 - Hollow fiber membrane for lipid peroxide adsorption and module using the same - Google Patents

Description

【００１９】
ポリスルホンの具体例としては、ユーデルポリスルホンＰ−１７００、Ｐ−３５００（テイジンアモコ社製）、ウルトラソンＳ３０１０、Ｓ６０１０（ＢＡＳＦ社製）、ビクトレックス（住友化学）、レーデルＡ（テイジンアモコ社製）、ウルトラソンＥ（ＢＡＳＦ社製）等のポリスルホンが挙げられる。又、本発明で用いられるポリスルホンは上記式（１）及び／又は（２）で表される繰り返し単位のみからなるポリマーが好適ではあるが、本発明の効果を妨げない範囲で他のモノマーと共重合していても良い。特に限定するものではないが、他の共重合モノマーは１０重量％以下であることが好ましい。
【００２０】
本発明においては、ポリビニルピロリドンを用いることが好ましい。ポリビニルピロリドンと前記支持体となる素材との構成・複合方法は特に限定されるものではなく、支持体となる素材とポリビニルピロリドンが積層されていても良いが、混合乃至は相溶されている方が好ましい。また、支持体となる素材としては特に限定されるものではないが、有機高分子ポリマーであることが好ましい。かかる有機高分子ポリマーとしてはポリスルホンが好ましい。
【００２１】
ポリビニルピロリドンとしては、市販されている重量平均分子量３６万、１６万、４万、１万のものが好適に用いられるが、もちろんそれ以外の分子量のものを使用してもかまわない。特に限定されるものではないが、ポリビニルピロリドンの重量平均分子量は２０００〜２００００００が好ましく、１００００〜１５０００００がより好ましい。なお、前記分子量は原料段階での分子量であり、最終製品においては、放射線架橋などにより分子量は前記値よりの遙かに大きなものとなっている場合もある。又、本発明で用いられるポリビニルピロリドンは、支持体となる素材とブレンドして用いる場合、ホモポリマーが好適ではあるが、本発明の効果を妨げない範囲で他のモノマーと共重合していても良い。特に限定するものではないが、他の共重合モノマーは１０重量％以下であることが好ましい。
【００２２】
なお、前記ポリビニルピロリドンや支持体となる素材（例えば、ポリスルホン）以外のポリマーなどが、本発明の効果を妨げない範囲で混合されていても良い。特に限定されるわけではないが、そのようなその他の素材は、中空糸膜全重量中、１０重量％以下であることが好ましい。
【００２３】
中空糸膜における膜厚は１０〜８０μｍが好ましく、２０〜６０μｍがより好ましい。又、膜の平均孔径については、１ｗｔ％アルブミン溶液におけるアルブミン透過率が０．５％以上であることが好ましく、１％以上がより好ましい。内径は１００〜８００μｍであることが好ましく、１５０〜３００μｍがより好ましい。また、本発明の中空糸膜は、人工腎臓としての機能を有することが好ましい。ここで、人工腎臓としての機能を有するというのは人工腎臓の機能分類でいうところのＩ型またはII型に属するものであることをいう（透析会誌、３２（１２）：１４６５〜１４６９，１９９９）。
【００２４】
本発明の中空糸膜は、従来知られている中空糸膜の製造方法などにおいて、前記したカチオンポリマーを導入する方法を適宜用いることにより製造される。中空糸膜の製法の一例として次のような方法がある。
【００２５】
ポリスルホンとポリビニルピロリドン（重量比率２０：１〜１：５が好ましく、５：１〜１：１がより好ましい）を良溶媒（Ｎ，Ｎ−ジメチルアセトアミド、ジメチルスルホキシド、ジメチルホルムアミド、Ｎ−メチルピロリドン、ジオキサンなどが好ましい）および良溶媒の混合溶液に溶解させた原液（濃度は、１０〜３０重量％が好ましく、１５〜２５重量％がより好ましい）を二重環状口金から吐出する際に内側に注入液を流し、乾式部を走行させた後凝固浴へ導く。この際、乾式部の湿度が影響を与えるために、乾式部走行中に膜外表面からの水分補給によって、外表面近傍での相分離挙動を速め、孔径拡大し、結果として透析の際の透過・拡散抵抗を減らすことも可能である。ただし、相対湿度が高すぎると外表面での原液凝固が支配的になり、かえって孔径が小さくなり、結果として透析の際の透過・拡散抵抗を増大する傾向がある。そのため、相対湿度としては６０〜９０％が好適である。また、注入液組成としてはプロセス適性から原液に用いた溶媒を基本とする組成からなるものを用いることが好ましい。注入液濃度としては、例えばジメチルアセトアミドを用いたときは、４５〜８０重量％、さらには６０〜７５重量％の水溶液が好適に用いられる。
【００２６】
カチオン性ポリマーを素材に含有させる方法には、素材の成形前の原液にカチオン性ポリマーを混和しておいて、成形する方法や、反応性を有する官能基を導入した素材を混和しておいて、素材を成形後、カチオン性ポリマーを官能基に対して表面反応により固定化する方法、通常の素材をカチオン性ポリマーの溶液に浸漬し、放射線照射、熱処理などによりカチオン性ポリマーを不溶化、固定化する方法、通常の素材にカチオン性ポリマーをコーティングし、吸着固定化する方法、さらに吸着固定化後、放射線照射、熱処理などにより不溶化、固定化する方法などが挙げられる。
【００２７】
また、カチオン性ポリマーを含有させるには膜内表面だけに固定化してもよいし、孔を有する膜全面に含有、固定化してもよい。
【００２８】
モジュールの製造方法としては、特に限定されないが、一例を示すと次の通りである。まず、中空糸膜を必要な長さに切断し、必要本数を束ねた後、筒状ケースに入れる。その後両端に仮のキャップをし、中空糸膜両端部にポッティング剤を入れる。このとき遠心機でモジュールを回転させながらポッティング剤を入れる方法は、ポッティング剤が均一に充填されるために好ましい方法である。ポッティング剤が固化した後、中空糸膜の両端が開口するように両端部を切断し、中空糸膜モジュールを得る。
【００２９】
本発明の中空糸膜は、体外循環用モジュールとして、人工腎臓、血漿分離等に好適に用いられる。人工腎臓に用いるような場合は、ＬＤＬやＨＤＬのような大分子は透析・濾過されない。従って、膜内表面に過酸化脂質と選択的に相互作用するようなリガンドを含有させてやれば、過酸化脂質は吸着除去され、ＬＤＬ、ＨＤＬは吸着されず、患者の体内に戻される。一方、血漿分離膜の場合には、ＬＤＬやＨＤＬは濾過されるが、膜全体に過酸化脂質と選択的に相互作用するようなリガンドを含有させてやれば、膜全体の面積を有効に使えるため過酸化脂質を効率よく除去するには有効である。ただし、濾過されたＬＤＬやＨＤＬは体内に戻すか、新たに新鮮血漿を補充することが好ましい。
【００３０】
以下、本発明の膜型吸着器の性能測定条件を記載する。
（１）抗酸化ＬＤＬ抗体の作製
板部らが作製したものを用いた（Ｈ．Ｉｔａｂｅ ｅｔ ａｌ．，Ｊ．Ｂｉｏｌ．Ｃｈｅｍ．２６９：１５２７４、１９９４）。すなわち、ヒト粥状硬化病巣ホモジェネートをマウスに注射して免疫、そのマウスの脾臓からハイブリドーマを作製し、硫酸銅処理ＬＤＬと反応するものを選別した。抗体クラスは、マウスＩｇＭで、未処理ＬＤＬ、アセチルＬＤＬ、マロンジアルデヒドＬＤＬとは反応しない。フォスファチジルコリンのアルデヒド誘導体やヒドロペルオキシドを含めていくつかのフォスファチジルコリン過酸化反応生成物と反応する。１５０ｍＭのＮａＣｌを含む１０ｍＭほう酸緩衝液（ｐＨ８．５）に溶解したものを用いた（蛋白濃度０．６０ｍｇ／ｍｌ）。
（２）酸化ＬＤＬの調製
市販のＬＤＬ（フナコシ製）を脱塩した後、０．２ｍｇ／ｍｌとなるようにリン酸緩衝液（ＰＢＳ）で希釈後、０．５ｍＭ硫酸銅水溶液を１ｗｔ％添加し、３７℃で１６時間反応させた。２５ｍＭのエチレンジアミン四酢酸（ＥＤＴＡ）を１ｗｔ％、１０ｗｔ％アジ化ナトリウムを０．０２ｗｔ％となるように添加したものを酸化ＬＤＬ標品とした。
（３）灌流操作
健常者血漿（日本人、３０歳、ＬＤＬ（βリポ蛋白）濃度２７５ｍｇ／ｄｌ，ＨＤＬ−コレステロール濃度７０ｍｇ／ｄｌ）に上記酸化ＬＤＬを２μｇ／ｍｌとなるように添加した。
【００３１】
内径２００μｍ、膜厚４０μｍの中空糸膜から、長さ１２ｃｍ、本数７０本のミニモジュール（内表面積５３ｃｍ²）を作成し、内径７ｍｍ（外径１０ｍｍ）、長さ２ｃｍのシリコーンチューブ（製品名ＡＲＡＭ）と異形コネクターを介して、内径０．８ｍｍ（外径１ｍｍ）のシリコーンチューブ（製品名ＡＲＡＭ、３７ｃｍのものを両端に２本）でつなぎ、上記血漿１．５ｍｌを０．５ｍｌ／分の流量で２５℃、４時間中空糸内に灌流した（素材表面積１ｍ²あたりの血漿量は２．８×１０²ｍｌ／ｍ²）。
【００３２】
さらにミニモジュールをつけずにシリコーンチューブのみで灌流操作も行った。
【００３３】
灌流前後の血漿中の酸化ＬＤＬ、ＬＤＬ、ＨＤＬ濃度を定量することにより、それぞれの吸着除去率を下記式により算出した。
【００３４】
吸着除去率（％）＝ミニモジュールでの吸着除去率（％）−シリコーンチューブのみでの吸着除去率（％）
それぞれの吸着除去率（％）＝１００×（灌流前の濃度−灌流後の濃度）／灌流前の濃度
（４）酸化ＬＤＬ、ＬＤＬ、ＨＤＬ濃度の測定
抗酸化ＬＤＬ抗体をＰＢＳで５μｇ／ｍｌに希釈し、９６穴のプレートに１００μｌ／ウェルずつ分注し、室温で２時間震盪した後、４℃にて一晩以上壁に吸着させた。
【００３５】
ウェル中の抗体溶液を捨て、１ｗｔ％Ｂｏｖｉｎｅ Ｓｅｒｕｍ Ａｌｂｍｉｎ（ＢＳＡ、フラクションＶ、生化学工業）を含むトリス−塩酸緩衝液（ｐＨ８．０）を２００μｌ／ウェルずつ分注し、室温で２時間震盪して壁をブロッキングした後、ウェル中のＢＳＡ溶液を捨て、酸化ＬＤＬを含んだ血漿および検量線作成用のスタンダード（０〜２μｇ／ｍｌの酸化ＬＤＬを含むＰＢＳ緩衝液）１００μｌ／ウェルずつ分注した。その後、室温で３０分震盪した後、４℃で一晩放置した。
【００３６】
室温に戻し、ウェル中の溶液を捨て、０．０５ｗｔ％トゥイーン−２０（片山化学）を含むトリス−塩酸緩衝液（ｐＨ８．０）でウェルを３回洗浄した。洗浄したウェルにＰＢＳで２０００倍に希釈したヒツジ抗アポＢ抗体（ＴＨＥ ＢＩＮＤＩＮＧ ＳＩＴＥ）１００μｌ／ウェルずつ分注し、室温で２時間震盪した後、ウェル中の抗アポＢ抗体を捨て、０．０５ｗｔ％トゥイーン−２０を含むトリス−塩酸緩衝液（ｐＨ８．０）でウェルを３回洗浄した。洗浄したウェルに２ｗｔ％ブロックエース（大日本製薬）を含むトリス−塩酸緩衝液（ｐＨ８．０）で２０００倍に希釈したアルカリ性フォスファターゼ標識ロバ抗ヒツジＩｇＧ抗体（ＣＨＥＭＩＣＯＮ）を１００μｌ／ウェルずつ分注し、室温で２時間震盪した。その後、ウェル中の標識抗体を捨て、０．０５ｗｔ％トゥイーン−２０を含むトリス−塩酸緩衝液（ｐＨ８．０）でウェルを３回洗浄し、さらにトリス−塩酸緩衝液（ｐＨ８．０）で２回洗浄した。続いて、ｐ−ニトロフェニルリン酸（Ｂｏｅｈｒｉｎｇｅｒ Ｍａｎｎｈｅｉｍ ＧｍｂＨ）の１ｍｇ／ｍｌ溶液（０．０００５Ｍ ＭｇＣｌ₂、１Ｍジエタノールアミン緩衝液、ｐＨ９．８）を１００μｌ／ウェルずつ分注し、適当な時間室温で反応させた後、４１５ｎｍの吸光度をプレートリーダーで測定した。スタンダードの結果から検量線を引き、酸化ＬＤＬ濃度を決定した。
【００３７】
ＬＤＬの定量はβ−リポ測定キット（和光純薬）を用いて行った。
【００３８】
ＨＤＬの定量はＨＤＬ−Ｃ測定キット（和光純薬）を用いて行った。
【００３９】
【実施例】
実施例
ポリスルホン（テイジンアモコ社製ユーデルＰ−３５００）１８重量部、ポリビニルピロリドン（ＢＡＳＦ社製Ｋ３０）９重量部をＮ，Ｎ−ジメチルアセトアミド７２重量部、水１重量部に加え、９０℃１４時間加熱溶解した。この製膜原液を外径０．３ｍｍ、内径０．２ｍｍのオリフィス型二重円筒型口金より吐出し芯液としてジメチルアセトアミド５８重量部、水４２重量部からなる溶液を吐出させ、乾式長３５０ｍｍを通過した後、水１００％の凝固浴に導き中空糸を得た。ポリスルホン中空糸分離膜を７０本束ね、中空糸中空部を閉塞しないようにエポキシ系ポッティング剤で両末端をガラス管モジュールケースに固定し、ミニモジュールを作成し、１ｗｔ％のポリエチレンイミン（分子量１万、和光純薬製）水溶液を充填して、２５ｋＧｙにてγ線照射した。該ミニモジュールの直径は約７ｍｍ、長さは１２ｃｍであった。放射線処理した膜は、再度蒸留水で３７℃で３０分間洗浄した後、吸着実験に供した。
【００４０】
実験結果は、表１に示した。
比較例１
実施例と同様にしてミニモジュールを作成し、１ｗｔ％のポリエチレンイミン水溶液の代わりに水を充填して、２５ｋＧｙにてγ線照射したものを作成した。
比較例２
市販のポリメチルメタクリレート透析膜（東レ、ＢＧ）を用いてミニモジュールを作成した。
比較例３
市販のビタミンＥコーティングセルロース膜（エクセブレン、テルモ、ＣＬ−Ｅ１５Ｎ）を用いてミニモジュールを作成した。
【００４１】
実施例および比較例１〜３における酸化ＬＤＬ、ＬＤＬ、ＨＤＬの吸着除去率の結果を表１に示した。
【００４２】
なお、実施例１で作成した中空糸膜の透水性およびβ₂ＭＧクリアランスは比較例１のものと差は見られなかった。
【００４３】
【表１】

【００４４】
【発明の効果】
本発明により、膜型血液浄化器などの用途に用いられ、特に透析患者において動脈硬化の進展を予防したり、その発症を未然に防ぐような場合に好適に用いられる過酸化等質吸着用中空糸膜を提供することができる。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a hollow fiber membrane for lipid peroxide adsorption and a module using the same. In particular, the present invention relates to a hollow fiber membrane that can be suitably used as a plasma separation membrane or an artificial kidney and can adsorb and remove lipid peroxide in blood.
[0002]
[Prior art]
Conventionally, hollow fiber membrane blood treatment devices using hollow fiber membranes have been widely used in the field of extracorporeal blood circulation, particularly in hemodialysis and plasma separation, and in recent years, particularly in the fields of dialysis membranes, blood component separation membranes and the like. A polymer hollow fiber membrane is widely used. However, it has been confirmed that patients with long-term hemodialysis have decreased blood antioxidant activity and high levels of lipid peroxide, which may be attributed to the arteries of long-term dialysis patients. The number of sclerosis diseases is increasing.
[0003]
Foam cells, which play an important role in the formation of atherosclerotic lesions, are the result of oxidatively altered lipids, especially low-density lipoproteins (hereinafter referred to as LDL), being taken up by macrophages, resulting in the foaming of macrophages. It is a thing.
[0004]
In order to solve these problems, a membrane-type blood purifier that coats the surface of a dialysis membrane with a coating of vitamin E having various physiological functions such as in vivo antioxidant action, biological membrane stabilization action, and platelet aggregation inhibition action Although it has been proposed (Japanese Patent Publication No. 62-41738), it is not possible to remove lipid peroxide in the blood of a patient once produced.
[0005]
Among lipid peroxides, oxidized LDL, in particular, has a variety of biological actions. In addition to the action of suppressing nitric oxide (NO) production from endothelial cells, monocytes are transported and accumulated in the subendothelium. Plays an important role in the onset and progression of arteriosclerosis, such as macrophages, taking oxidized LDL itself into foam cells, promoting plaque formation on the arterial wall, and promoting endothelial cell and smooth muscle cell damage. Yes. Therefore, it is desirable to remove lipid peroxide, particularly oxidized LDL, from the blood.
[0006]
Since patients with hyperlipidemia have a large amount of LDL in the blood, removal of LDL is effective in preventing the progression of arteriosclerosis. Thus, Kaneka Chemical's liposorber is currently used as an adsorption bead column for LDL. Oxidized LDL is also considered to be removed at the same time by using this adsorbent. However, in the case of dialysis patients, the LDL concentration in the blood is the same level as that of healthy people in dialysis patients. Therefore, it cannot be used for dialysis patients.
[0007]
Moreover, since high density lipoprotein (HDL) has a function as an arteriosclerosis protective factor, the HDL concentration in the blood of a dialysis patient should not be lowered.
[0008]
Lipid peroxide cannot be removed with dialysis membranes such as cellulose membranes, polymethyl methacrylate membranes, ethylene-vinyl alcohol membranes, polysulfone / polyvinylpyrrolidone blend membranes that have been marketed so far. Rather, the value of lipid peroxide in the blood tends to increase after dialysis due to stimulation by the dialysis membrane (kidney and dialysis separate volume, 38: 125 (1995)).
[0009]
[Problems to be solved by the invention]
An object of the present invention is to improve the drawbacks of the prior art, and in particular to provide a hollow fiber membrane capable of selectively adsorbing and removing lipid peroxide and a module using the same.
[0010]
[Means for Solving the Problems]
In order to solve the above problems, the present invention has the following configuration.
“A hollow fiber membrane for lipid peroxide adsorption, characterized in that the hollow fiber membrane material contains a cationic polymer.”
[0011]
DETAILED DESCRIPTION OF THE INVENTION
The hollow fiber membrane of the present invention is characterized in that the hollow fiber membrane material contains a cationic polymer. Cationic polymers include primary, secondary, tertiary amino groups, amino group-containing polymers having a quaternary ammonium salt, polymers having an aziridine (ethyleneimine) compound, chitin, chitosan, acrylamide polymers, and copolymers thereof. Examples thereof include a polymer, a nonion, and a copolymer with an anionic compound. The polymer may be linear, branched or cyclic. The molecular weight refers to a compound having 600 or more.
[0012]
Examples of amino group-containing polymers include polyalkyleneimines, polyallylamines, polyvinylamines, those having substituents introduced therein, and copolymers composed of these monomer units.
[0013]
Examples of the polyethyleneimine derivative include those obtained by derivatizing polyethyleneimine at various ratios such as alkylation, carboxylation, phenylation, phosphorylation, and sulfonation.
[0014]
As the polyethyleneimine, a linear or branched one having a molecular weight of 600 or more is used.
[0015]
Of the cationic polymers, branched polyethyleneimine is preferably used because of its low toxicity, availability, and ease of handling.
[0016]
The hollow fiber membrane material of the present invention is not particularly limited, but a material used for medical purposes is preferable, and examples thereof include polyvinyl chloride, cellulosic polymer, polystyrene, polymethyl methacrylate, polycarbonate, polysulfone, and polyurethane. . Among these, polysulfone is particularly suitable because it is easy to mold and has excellent membrane performance when formed into a membrane.
[0017]
The polysulfone used in the present invention has an aromatic ring, sulfonyl group and ether group in the main chain. For example, polysulfone represented by the chemical formulas of the following formulas (1) and (2) is preferably used. The invention is not limited to these. N in the formula is an integer such as 50 to 80.
[0018]
[Chemical 1]

[0019]
Specific examples of polysulfone include Udel polysulfone P-1700, P-3500 (manufactured by Teijin Amoco), Ultrason S3010, S6010 (manufactured by BASF), Victrex (Sumitomo Chemical), Radel A (manufactured by Teijin Amoco). And polysulfone such as Ultrason E (manufactured by BASF). In addition, the polysulfone used in the present invention is preferably a polymer composed only of the repeating unit represented by the above formula (1) and / or (2). However, it does not interfere with the effects of the present invention. It may be polymerized. Although it does not specifically limit, it is preferable that another copolymerization monomer is 10 weight% or less.
[0020]
In the present invention, it is preferable to use polyvinylpyrrolidone. The constitution / combination method of polyvinyl pyrrolidone and the material to be the support is not particularly limited, and the material to be the support and polyvinyl pyrrolidone may be laminated, but they are mixed or compatible with each other. Is preferred. Moreover, although it does not specifically limit as a raw material used as a support body, It is preferable that it is an organic high molecular polymer. Such an organic polymer is preferably polysulfone.
[0021]
As the polyvinyl pyrrolidone, commercially available ones having a weight average molecular weight of 360,000, 160,000, 40,000 and 10,000 are preferably used. Of course, other molecular weights may be used. Although not particularly limited, the weight average molecular weight of polyvinylpyrrolidone is preferably from 2,000 to 2,000,000, and more preferably from 10,000 to 1,000,000. The molecular weight is the molecular weight at the raw material stage, and in the final product, the molecular weight may be much larger than the above value due to radiation crosslinking or the like. In addition, the polyvinylpyrrolidone used in the present invention is preferably a homopolymer when blended with a material to be a support, but may be copolymerized with other monomers within a range that does not interfere with the effects of the present invention. good. Although it does not specifically limit, it is preferable that another copolymerization monomer is 10 weight% or less.
[0022]
In addition, polymers other than the polyvinyl pyrrolidone and the material (for example, polysulfone) used as a support may be mixed within a range that does not hinder the effects of the present invention. Although not particularly limited, such other materials are preferably 10% by weight or less based on the total weight of the hollow fiber membrane.
[0023]
The thickness of the hollow fiber membrane is preferably 10 to 80 μm, and more preferably 20 to 60 μm. The average pore diameter of the membrane is preferably such that the albumin permeability in a 1 wt% albumin solution is 0.5% or more, and more preferably 1% or more. The inner diameter is preferably 100 to 800 μm, more preferably 150 to 300 μm. The hollow fiber membrane of the present invention preferably has a function as an artificial kidney. Here, having the function as an artificial kidney means that it belongs to the type I or type II in the functional classification of the artificial kidney (Dialysis Journal, 32 (12): 1465-1469, 1999). .
[0024]
The hollow fiber membrane of the present invention is produced by appropriately using the above-described method for introducing a cationic polymer in a conventionally known method for producing a hollow fiber membrane. An example of a method for producing a hollow fiber membrane is as follows.
[0025]
Polysulfone and polyvinylpyrrolidone (weight ratio of 20: 1 to 1: 5 is preferable, and 5: 1 to 1: 1 is more preferable). Good solvents (N, N-dimethylacetamide, dimethylsulfoxide, dimethylformamide, N-methylpyrrolidone, Dioxane or the like is preferable) and a stock solution (concentration is preferably 10 to 30% by weight, more preferably 15 to 25% by weight) dissolved in a mixed solution of a good solvent and injected into the inside when discharging from the double annular die Pour the liquid, run the dry section, and then lead to the coagulation bath. At this time, the humidity of the dry part has an effect, so that the phase separation behavior near the outer surface is accelerated by replenishing moisture from the outer surface of the membrane while the dry part is running.・ Diffusion resistance can be reduced. However, when the relative humidity is too high, the solid solution coagulation on the outer surface becomes dominant, and the pore diameter becomes rather small, and as a result, there is a tendency to increase permeation / diffusion resistance during dialysis. Therefore, the relative humidity is preferably 60 to 90%. Moreover, it is preferable to use what consists of a composition based on the solvent used for the undiluted | stock solution as an injection | pouring liquid composition from process suitability. For example, when dimethylacetamide is used as an injection solution concentration, an aqueous solution of 45 to 80% by weight, more preferably 60 to 75% by weight, is preferably used.
[0026]
For the method of incorporating the cationic polymer into the material, the cationic polymer is mixed in the stock solution before molding the material, and the molding method or the material into which the functional group having reactivity is introduced is mixed. After molding the material, the cationic polymer is immobilized on the functional group by surface reaction. Ordinary materials are immersed in the cationic polymer solution, and the cationic polymer is insolubilized and immobilized by irradiation, heat treatment, etc. And a method of coating an ordinary material with a cationic polymer and adsorbing and immobilizing it, and further adsorbing and immobilizing and then insolubilizing and immobilizing by radiation irradiation, heat treatment and the like.
[0027]
In order to contain the cationic polymer, it may be immobilized only on the inner surface of the membrane, or may be contained and immobilized on the entire membrane having pores.
[0028]
Although it does not specifically limit as a manufacturing method of a module, It will be as follows if an example is shown. First, the hollow fiber membrane is cut to a required length, bundled in a necessary number, and then put into a cylindrical case. Then, a temporary cap is put on both ends, and a potting agent is put on both ends of the hollow fiber membrane. At this time, the method of adding the potting agent while rotating the module with a centrifuge is a preferable method because the potting agent is uniformly filled. After the potting agent is solidified, both ends are cut so that both ends of the hollow fiber membrane are open, and a hollow fiber membrane module is obtained.
[0029]
The hollow fiber membrane of the present invention is suitably used as an extracorporeal circulation module for artificial kidneys, plasma separation, and the like. When used for an artificial kidney, large molecules such as LDL and HDL are not dialyzed and filtered. Accordingly, if a ligand capable of selectively interacting with lipid peroxide is contained on the inner surface of the membrane, the lipid peroxide is adsorbed and removed, and LDL and HDL are not adsorbed and returned to the patient's body. On the other hand, in the case of a plasma separation membrane, LDL and HDL are filtered, but if the whole membrane contains a ligand that selectively interacts with lipid peroxide, the entire membrane area can be used effectively. Therefore, it is effective for removing lipid peroxide efficiently. However, it is preferable that the filtered LDL or HDL is returned to the body or newly supplemented with fresh plasma.
[0030]
Hereinafter, the performance measurement conditions of the membrane type adsorber of the present invention will be described.
(1) Production of Antioxidant LDL Antibody The one produced by Plate et al. Was used (H. Itabet et al., J. Biol. Chem. 269: 15274, 1994). That is, human atherosclerotic lesion homogenate was injected into mice for immunization, hybridomas were prepared from the spleens of the mice, and those that reacted with copper sulfate-treated LDL were selected. The antibody class is mouse IgM and does not react with untreated LDL, acetyl LDL, or malondialdehyde LDL. Reacts with several phosphatidylcholine peroxidation products, including aldehyde derivatives of phosphatidylcholine and hydroperoxides. What was dissolved in 10 mM borate buffer (pH 8.5) containing 150 mM NaCl was used (protein concentration 0.60 mg / ml).
(2) Preparation of oxidized LDL After desalting commercially available LDL (manufactured by Funakoshi), diluted with a phosphate buffer solution (PBS) to a concentration of 0.2 mg / ml, 1 wt% of 0.5 mM aqueous copper sulfate solution was added. And reacted at 37 ° C. for 16 hours. What added 25 mM ethylenediaminetetraacetic acid (EDTA) so that 1 wt% and 10 wt% sodium azide might become 0.02 wt% was made into the oxidation LDL sample.
(3) Perfusion operation The above-mentioned oxidized LDL was added to plasma (Japanese, 30 years old, LDL (β-lipoprotein) concentration 275 mg / dl, HDL-cholesterol concentration 70 mg / dl) at 2 μg / ml.
[0031]
From a hollow fiber membrane having an inner diameter of 200 μm and a film thickness of 40 μm, a mini-module (inner surface area 53 cm ² ) having a length of 12 cm and a number of 70 is prepared, and a silicone tube (product name ARAM) having an inner diameter of 7 mm (outer diameter of 10 mm) ) And a deformed connector, connected by a 0.8 mm inner diameter (outside diameter 1 mm) silicone tube (product name: ARAM, two 37 cm ones at both ends), and 1.5 ml of the plasma flowed at 0.5 ml / min At 25 ° C. for 4 hours in the hollow fiber (the plasma volume per 1 m ^{2 of the} material surface area was 2.8 × 10 ² ml / m ² ).
[0032]
Furthermore, the perfusion operation was also performed only with the silicone tube without attaching the mini module.
[0033]
By quantifying the oxidized LDL, LDL, and HDL concentrations in plasma before and after perfusion, each adsorption removal rate was calculated by the following equation.
[0034]
Adsorption removal rate (%) = Adsorption removal rate with mini module (%)-Adsorption removal rate with silicone tube only (%)
Each adsorption removal rate (%) = 100 × (concentration before perfusion−concentration after perfusion) / concentration before perfusion (4) Measurement of oxidized LDL, LDL and HDL concentrations Antioxidant LDL antibody was adjusted to 5 μg / ml with PBS. Diluted, dispensed into a 96-well plate at 100 μl / well, shaken at room temperature for 2 hours, and adsorbed to the wall at 4 ° C. overnight.
[0035]
Discard the antibody solution in the well, dispense 200 μl / well of Tris-HCl buffer (pH 8.0) containing 1 wt% Bovine Serum Albumin (BSA, Fraction V, Seikagaku), and shake at room temperature for 2 hours. After blocking the wall, the BSA solution in the well was discarded, and 100 μl / well of plasma containing oxidized LDL and a standard for preparing a calibration curve (PBS buffer containing 0-2 μg / ml oxidized LDL) were dispensed. . Then, after shaking at room temperature for 30 minutes, it was left at 4 ° C. overnight.
[0036]
After returning to room temperature, the solution in the well was discarded, and the well was washed three times with Tris-HCl buffer (pH 8.0) containing 0.05 wt% Tween-20 (Katayama Chemical). 100 μl / well of sheep anti-apo B antibody (THE BINDING SITE) diluted 2000-fold with PBS to the washed wells and shaken at room temperature for 2 hours, the anti-apo B antibody in the wells was discarded, and 0.05 wt. The wells were washed 3 times with Tris-HCl buffer (pH 8.0) containing% Tween-20. 100 μl / well of alkaline phosphatase-labeled donkey anti-sheep IgG antibody (CHEMICON) diluted 2000 times with Tris-HCl buffer (pH 8.0) containing 2 wt% Block Ace (Dainippon Pharmaceutical Co., Ltd.) was dispensed into the washed wells. Shake for 2 hours at room temperature. Thereafter, the labeled antibody in the wells is discarded, and the wells are washed three times with Tris-HCl buffer (pH 8.0) containing 0.05 wt% Tween-20, and further 2 with Tris-HCl buffer (pH 8.0). Washed twice. Subsequently, a 1 mg / ml solution (0.0005M MgCl ₂ , 1M diethanolamine buffer, pH 9.8) of p-nitrophenyl phosphate (Boehringer Mannheim GmbH) was dispensed at 100 μl / well and reacted at room temperature for an appropriate time. Then, the absorbance at 415 nm was measured with a plate reader. A calibration curve was drawn from the standard results to determine the oxidized LDL concentration.
[0037]
LDL was quantified using a β-lipo assay kit (Wako Pure Chemical Industries).
[0038]
Quantification of HDL was performed using an HDL-C measurement kit (Wako Pure Chemical Industries).
[0039]
【Example】
Example: Polysulfone (Udel P-3500 manufactured by Teijin Amoco) 18 parts by weight, polyvinylpyrrolidone (BASF K30) 9 parts by weight were added to 72 parts by weight of N, N-dimethylacetamide and 1 part by weight of water, and 90 ° C. for 14 hours. Dissolved by heating. This film-forming stock solution is discharged from an orifice type double cylindrical die having an outer diameter of 0.3 mm and an inner diameter of 0.2 mm, and a solution consisting of 58 parts by weight of dimethylacetamide and 42 parts by weight of water is discharged as a core liquid. After passing, the hollow fiber was obtained by being led to a coagulation bath with 100% water. 70 polysulfone hollow fiber separation membranes are bundled, both ends are fixed to a glass tube module case with an epoxy potting agent so as not to block the hollow portion of the hollow fiber, a mini-module is prepared, and 1 wt% polyethyleneimine (molecular weight 10,000) , Manufactured by Wako Pure Chemical Industries, Ltd.) and filled with an aqueous solution and irradiated with γ rays at 25 kGy. The mini module had a diameter of about 7 mm and a length of 12 cm. The membrane treated with radiation was again washed with distilled water at 37 ° C. for 30 minutes and then subjected to an adsorption experiment.
[0040]
The experimental results are shown in Table 1.
Comparative Example 1
Mini-modules were prepared in the same manner as in the Examples, and water was filled in place of the 1 wt% polyethyleneimine aqueous solution and irradiated with γ rays at 25 kGy.
Comparative Example 2
A mini-module was prepared using a commercially available polymethylmethacrylate dialysis membrane (Toray, BG).
Comparative Example 3
Minimodules were made using commercially available vitamin E-coated cellulose membranes (Exbrelen, Terumo, CL-E15N).
[0041]
Table 1 shows the results of adsorption removal rates of oxidized LDL, LDL, and HDL in Examples and Comparative Examples 1 to 3.
[0042]
In addition, the water permeability and β ₂ MG clearance of the hollow fiber membrane prepared in Example 1 were not different from those of Comparative Example 1.
[0043]
[Table 1]

[0044]
【The invention's effect】
According to the present invention, it is used for applications such as a membrane blood purifier, and is particularly suitable for use in a case where the progression of arteriosclerosis is prevented or prevented in advance in dialysis patients. A yarn membrane can be provided.

Claims (9)

A. The containing polyvinylpyrrolidone material obtained by introducing a functional group having reactivity was formed by mixing hollow fiber membrane materials, a method of fixing by surface reaction against polyethyleneimine and derivatives thereof to said functional group, B. The hollow fiber membrane materials containing polyvinyl pyrrolidone was immersed in a solution of polyethylene imine and its derivatives, a method for immobilizing the polyethyleneimine and derivatives thereof by radiation or heat treatment and C. After coating the polyethyleneimine and its derivatives in the hollow fiber membrane materials containing polyvinylpyrrolidone, by irradiation or heat treatment polyethyleneimine and derivatives thereof are contained in the hollow fiber membrane material by any method of a method of immobilizing A hollow fiber membrane for adsorbing lipid peroxide, characterized by comprising: From a hollow fiber membrane having an inner diameter of 200 μm and a film thickness of 40 μm, a mini module (inner surface area 53 cm ² ) having a length of 12 cm and a number of 70 is prepared, and a silicone tube and a deformed connector having an inner diameter of 7 mm (outer diameter of 10 mm) and a length of 2 cm. Through a silicone tube with an inner diameter of 0.8 mm (outer diameter of 1 mm) (two 37 cm ones at each end) and 1.5 ml of plasma at a flow rate of 0.5 ml / min at 25 ° C. for 4 hours in the hollow fiber. When the perfusion operation is performed under the condition that the amount of plasma per 1 m ^{2 of} the material flowing is 2.8 × 10 ² ml / m ² , the oxidized low density with an initial concentration of 2 μg / ml contained in the plasma 2. The hollow fiber membrane for adsorbing lipid peroxide according to claim 1, wherein the adsorption removal rate of lipoprotein is 20% or more. The hollow fiber membrane for adsorbing lipid peroxides according to claim 1 or 2 , wherein the adsorption removal rate of low density lipoprotein is less than 5%. The hollow fiber membrane for lipid peroxide adsorption according to any one of claims 1 to 3 , wherein the adsorption removal rate of high-density lipoprotein is less than 5%. The hollow fiber membrane for lipid peroxide adsorption according to any one of claims 1 to 4 , wherein the hollow fiber membrane material is made of polysulfone. Hollow fiber membranes Hollow fiber membranes for lipid peroxide adsorption according to any one of claims 1 to 5, characterized in that used in module for extracorporeal circulation. The hollow fiber membrane for lipid peroxide adsorption according to claim 6 , wherein the extracorporeal circulation module has a function as a plasma separation membrane. 7. The lipid peroxide-adsorbing hollow fiber membrane according to claim 6 , wherein the extracorporeal circulation module has a function as an artificial kidney. A module comprising the hollow fiber membrane for adsorbing lipid peroxide according to any one of claims 1 to 8 .