JP4783971B2 - Adsorbent for adsorption of oxidized low density lipoprotein - Google Patents

Description

【００１６】
ポリスルホンの具体例としては、”ユーデル”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。また、本発明で用いられるポリスルホンは上記式（１）および／または（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。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。
【００１７】
本発明においては、支持体の血液適合性を向上させるために、親水性高分子をブレンドすることが好ましい。親水性高分子としては、ポリエチレングリコール、ポリビニルピロリドン等が、血液適合性の点で好ましい。また、親水性高分子については、カチオン性ポリマーと放射線架橋できるものであるとより好ましい。ポリスルホンとの相溶性から、特にポリビニルピロリドンが好ましい。ポリビニルピロリドンは、支持体となる素材と積層されていても良いが、混合ないしは相溶されている方が好ましい。
【００１８】
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。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遙かに大きなものとなっている場合もある。また、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。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。
【００１９】
なお、前記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が、そのようなその他の素材は、支持体全重量中、１０重量％以下であることが好ましい。
【００２０】
本発明の吸着材は、体外循環カラム等に充填し、体外循環治療に用いることができる。すなわち、中空糸膜状、繊維状、ビーズ状などにして用いることができる。特に人工腎臓用の中空糸膜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付与させてやれば、尿毒素や水分などの除去という従来の機能に加え、ＬＤＬや高密度リポ蛋白（以下ＨＤＬという）などをほとんど損なわずに、体液中の酸化ＬＤＬ除去するという新たな機能を付加させることもできる。とりわけ、長期に血液透析を行っている患者の中には、血中抗酸化作用の低下や過酸化脂質が高値であることが確認されており、これに起因すると思われる長期透析患者の動脈硬化性疾患等が増加しているため、透析患者の体液中の酸化ＬＤＬを除去する目的で本発明の吸着材を用いることは好ましい。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ジアルキルアミノアルキルデキストランを付与させてやれば、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。
【００２１】
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（以下ＰＢＳという）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（以下ＥＤＴＡという）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）吸着実験操作
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。
【００２２】
表面積９ｃｍ²の支持体を上記血漿０．３ｍｌ中に３７℃、４時間静置した。（支持体表面積１ｍ²あたりの血漿量は３．３×１０²ｍｌ／ｍ²）。
【００２３】
膜と相互作用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。
【００２４】
それぞれの吸着除去率（％）＝１００×（相互作用前の濃度−相互作用後の濃度）／相互作用前の濃度
（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。
【００２５】
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）を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。
【００２６】
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）を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。
【００２７】
ＬＤＬの定量はβ−リポ測定キット（和光純薬）を用いて行った。
【００２８】
ＨＤＬの定量はＨＤＬ−Ｃ測定キット（和光純薬）を用いて行った。
【００２９】
【実施例】
以下、実施例により本発明をさらに具体的に説明するが、本発明はこのような実施例に限定されるものではない。
（１）支持体の作成
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Ｋ９０）６重量部、ポリビニルピロリドン（ＢＡＳＦ社製Ｋ３０）３重量部、Ｎ，Ｎ−ジメチルアセトアミド７２重量部、水１重量部の溶液を調製した。その後、この溶液を０．１ｍｍスペーサー付きのガラス板に塗布し、ドクターブレードにて膜になるよう引き延ばした後すぐに、Ｎ，Ｎ−ジメチルアセトアミド５２重量部、水４８重量部に調製した溶液に、ガラス板を浸漬し、膜を凝固させた。一連の操作の操作はすべて５０℃で行った。
（２）ジエチルアミノエチルデキストランの支持体への固定化
１ｗｔ％のジエチルアミノエチルデキストラン（ＳＩＧＭＡ社製分子量５０万）水溶液を調製した。（１）で作成したポリビニルピロリドンブレンドポリスルホン平膜をエチルアミノエチルデキストラン水溶液に浸漬した状態で、２５ｋＧｙのγ線を照射することでジエチルアミノエチルデキストランを固定化した。その後、５０℃温水中で１晩放置することで、膜に固定化されていないジエチルアミノエチルデキストランを抽出、洗浄した。
【００３０】
実験結果は、表１に示した。
比較例１
実施例と同様にしてポリスルホン／ポリビニルピロリドン平膜を作成し、１ｗｔ％のジエチルアミノエチルデキストラン水溶液の代わりに水を充填して、２５ｋＧｙのγ線を照射したものを作成した。
比較例２
実施例と同様にしてデキストラン（ＳＩＧＭＡ社製分子量５０万）を固定化したポリスルホン／ポリビニルピロリドン平膜を作成した。
【００３１】
実施例および比較例１、２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。
【００３２】
【表１】

【００３３】
【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吸着材を提供することができる。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to an adsorbent for oxidized low density lipoprotein (hereinafter referred to as oxidized LDL). More specifically, the present invention relates to an adsorbent for selectively removing oxidized LDL from body fluids and suppressing reduction or progression of various symptoms of arteriosclerosis.
[0002]
[Prior art]
Arteriosclerosis is a general term for lesions that show thickening of the arterial wall and have lost elasticity and have become hardened. Among them, the frequency is extremely high and the important disease is atherosclerosis, Generally speaking, arteriosclerosis often means atherosclerosis. It is lipid peroxide that plays an important role in the formation of atherosclerotic lesions. Among them, oxidized LDL has various biological actions, and in addition to the action of suppressing nitric oxide (NO) production from endothelial cells, monocytes are transported and accumulated in the subendothelium, and they themselves become macrophages. It plays an important role in the onset and progression of arteriosclerosis by taking oxidized LDL itself into foam cells and promoting plaque formation on the arterial wall, as well as promoting endothelial cell and smooth muscle cell damage. Therefore, it is desirable to remove lipid peroxide, particularly oxidized LDL, from the blood.
[0003]
Until now, the only methods for treating arteriosclerosis have been to spread surgically constricted blood vessels or to inhibit the metabolism of LDL (hereinafter referred to as LDL) using drugs. The former is highly invasive and cannot prevent the progression of arteriosclerosis. The latter has a problem that it is not effective for arteriosclerosis patients without LDL metabolic abnormality.
[0004]
[Problems to be solved by the invention]
The present invention provides an adsorbent capable of selectively adsorbing oxidized LDL with almost no loss of active ingredients in body fluids in order to solve the above problems.
[0005]
[Means for Solving the Problems]
In order to solve the above problems, the present invention has the following configuration. “A lipid peroxide adsorbent characterized in that a dialkylaminoalkyldextran is added to a support.”
[0006]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, the present invention will be described in more detail.
[0007]
The adsorbent of the present invention is characterized in that a dialkylaminoalkyldextran having a selective affinity for oxidized LDL is imparted to a support. Moreover, the copolymer which consists of the monomer unit which comprises dialkylaminoalkyl dextran may be sufficient. In addition, diethylaminoethyldextran is preferably used because of its availability. In addition, diethylaminoethyl dextran is preferable from the viewpoint of safety because it has been used in pharmaceuticals (F. Kazuya et al., USpat. 3,851,057 (1974 to Meito Sangyo)).
[0008]
In the case of hyperlipidemia patients, the amount of LDL in the blood is large, so removing LDL is effective in preventing the progression of arteriosclerosis, but in the case of dialysis patients and patients with heart disease, The LDL concentration in the medium is often at the same level as healthy individuals. Moreover, since high-density lipoprotein (HDL) has a function as an arteriosclerosis protective factor, the HDL concentration in the blood of dialysis patients and patients with heart disease should not be lowered. Thus, it is desirable to selectively remove lipid peroxide, particularly oxidized LDL, from dialysis patients and patients with heart disease. In the present invention, when the adsorbent and plasma interact with each other under the condition that the plasma volume per 1 m ^{2 of} the support surface area is 3.3 × 10 ² ml / m ² , the initial amount contained in the plasma The adsorption removal rate of oxidized low density lipoprotein with a concentration of 2 μg / ml is 60% or more, the adsorption removal rate of low density lipoprotein is less than 20%, and the adsorption removal rate of high density lipoprotein is less than 15%. It can be used as an index of selectivity.
[0009]
The support in the present invention refers to a substance that does not dissolve in water for immobilizing dialkylaminoalkyldextran. The support used in the present invention may be either organic or inorganic. Specific examples of the material of the support include aromatic polyvinyl compounds represented by polystyrene, polysulfone, polyetherimide, polyamide, polyimide, polyether, polyphenylene sulfide, and the like, but other body fluid components other than the target oxidized LDL Polysulfone which is difficult to adsorb is preferred. The term “body fluid” as used herein refers to blood, ascites, lymph fluid, intra-articular fluid, other biological components derived from living bodies, and fractional components obtained therefrom.
[0010]
The adsorbent of the present invention is characterized by imparting dialkylaminoalkyldextran to a support. As the imparting method, a physical adsorption method, an ionic bond method, a covalent bond method, or the like can be used. Specifically, a method in which dialkylaminoalkyldextran is mixed with a stock solution before molding of the support and molding, or a material into which a functional group having reactivity is introduced is mixed with the support and the support is used. A method of immobilizing dialkylaminoalkyl dextran to a functional group by surface reaction after molding, a method in which a normal support is immersed in a dialkylaminoalkyl dextran solution, and insolubilizing dialkylaminoalkyl dextran by irradiation, heat treatment, etc. Examples thereof include a method of coating a support with dialkylaminoalkyldextran and adsorbing and immobilizing, and a method of insolubilizing by irradiation and heat treatment after adsorbing and immobilizing.
[0011]
The shape of the support may be any shape such as a flat membrane, a hollow fiber membrane, a fibrous shape, and a granular shape.
[0012]
Further, in order to apply the dialkylaminoalkyldextran to the support, it may be immobilized only on the inner surface of the film, or may be applied to the entire film having pores.
[0013]
When the material of the support is a polymer compound, the molecular weight is preferably 5000 or more and 1,000,000 or less, particularly 10,000 or more and 200,000 or less.
[0014]
In particular, when the material of the support is polysulfone, those having an aromatic ring, sulfonyl group and ether group in the main chain, for example, polysulfone represented by the following chemical formula are preferably used. It is not limited. N in the formula is an integer of 50 to 80.
[0015]
[Chemical 1]

[0016]
Specific examples of polysulfone include “Udel” P-1700, P-3500 (manufactured by Teijin Amoco), “Ultrason” S3010, S6010 (manufactured by BASF), “Victrex” (Sumitomo Chemical), “Radel” A Examples include polysulfones such as (manufactured by Teijin Amoco) and "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). It may be polymerized. The other copolymerization monomer is preferably 10% by weight or less.
[0017]
In the present invention, it is preferable to blend a hydrophilic polymer in order to improve blood compatibility of the support. As the hydrophilic polymer, polyethylene glycol, polyvinyl pyrrolidone and the like are preferable in terms of blood compatibility. The hydrophilic polymer is more preferably one that can be radiation-crosslinked with the cationic polymer. In view of compatibility with polysulfone, polyvinylpyrrolidone is particularly preferable. Polyvinyl pyrrolidone may be laminated with a material to be a support, but is preferably mixed or compatible.
[0018]
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. 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 a 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 as long as the effects of the present invention are not hindered. good. The other copolymerization monomer is preferably 10% by weight or less.
[0019]
In addition, although polymers other than the above may be mixed in the range which does not prevent the effect of this invention, it is preferable that such other raw materials are 10 weight% or less in the support total weight.
[0020]
The adsorbent of the present invention can be packed in an extracorporeal circulation column or the like and used for extracorporeal circulation treatment. That is, it can be used in the form of a hollow fiber membrane, fiber, bead or the like. In particular, if a hollow fiber membrane for artificial kidneys is given a ligand that selectively interacts with lipid peroxide, in addition to the conventional function of removing uremic toxins and water, LDL and high-density lipoprotein ( It is also possible to add a new function of removing oxidized LDL from body fluids with little loss of HDL). In particular, it has been confirmed that patients with long-term hemodialysis have low blood antioxidant activity and high levels of lipid peroxide, which may be attributed to arteriosclerosis in long-term dialysis patients. Since sexual diseases and the like are increasing, it is preferable to use the adsorbent of the present invention for the purpose of removing oxidized LDL in the body fluid of dialysis patients. On the other hand, in the case of a plasma separation membrane, LDL and HDL are filtered, but if the dialkylaminoalkyldextran that selectively interacts with lipid peroxide is added to the entire membrane, the area of the entire membrane is effectively increased. It can be used effectively to remove lipid peroxide efficiently. However, it is preferable that the filtered LDL or HDL is returned to the body or newly supplemented with fresh plasma.
[0021]
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 Commercially available LDL (manufactured by Funakoshi) was desalted and diluted with a phosphate buffer (hereinafter referred to as PBS) to a concentration of 0.2 mg / ml. % Was added and reacted at 37 ° C. for 16 hours. An oxidized LDL preparation was prepared by adding 25 mM ethylenediaminetetraacetic acid (hereinafter referred to as EDTA) to a concentration of 1 wt%, 10 wt% sodium azide to 0.02 wt%.
(3) Operation of adsorption experiment The above-mentioned oxidized LDL was added to healthy human plasma (Japanese, 30 years old, LDL (β-lipoprotein) concentration 275 mg / dl, HDL-cholesterol concentration 70 mg / dl) to 2 μg / ml.
[0022]
A support having a surface area of 9 cm ² was allowed to stand in 0.3 ml of the plasma at 37 ° C. for 4 hours. (The plasma volume per 1 m ^{2 of the} support surface area is 3.3 × 10 ² ml / m ² ).
[0023]
By quantifying the concentrations of oxidized LDL, LDL, and HDL in the plasma before and after interaction with the membrane, the respective adsorption removal rates were calculated by the following formula.
[0024]
Each adsorption removal rate (%) = 100 × (concentration before interaction−concentration after interaction) / concentration before interaction (4) Measurement of oxidized LDL, LDL, HDL concentration Antioxidant LDL antibody in PBS at 5 μg The solution was diluted to 100 ml / well in a 96-well plate, shaken at room temperature for 2 hours, and adsorbed on the wall at 4 ° C overnight or longer.
[0025]
Discard the antibody solution in the well and dispense 200 μl / well of Tris-HCl buffer (pH 8.0) containing 1 wt% Bovine Serum Albumin (BSA, “Fraction V”, Seikagaku Corporation) for 2 hours at room temperature. After blocking the wall by shaking, the BSA solution in the well is discarded, and plasma containing oxidized LDL and a standard for preparing a calibration curve (PBS buffer containing 0-2 μg / ml oxidized LDL) are added at 100 μl / well. Dispensed. Then, after shaking at room temperature for 30 minutes, it was left at 4 ° C. overnight.
[0026]
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 times with PBS was dispensed into the washed wells and shaken at room temperature for 2 hours, and then the anti-apo B antibody in the wells was discarded. The wells were washed three times with Tris-HCl buffer (pH 8.0) containing 05 wt% “Tween-20”. 100 μl / well of alkaline phosphatase-labeled donkey anti-sheep IgG antibody (CHEMICON) diluted 2000-fold with Tris-HCl buffer (pH 8.0) containing 2 wt% “Block Ace” (Dainippon Pharmaceutical) in the washed wells Poured and shaken for 2 hours at room temperature. Thereafter, the labeled antibody in the well is discarded, and the well is washed three times with Tris-HCl buffer (pH 8.0) containing 0.05 wt% “Tween-20”, and further, Tris-HCl buffer (pH 8.0). And 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.
[0027]
LDL was quantified using a β-lipo assay kit (Wako Pure Chemical Industries).
[0028]
Quantification of HDL was performed using an HDL-C measurement kit (Wako Pure Chemical Industries).
[0029]
【Example】
EXAMPLES Hereinafter, although an Example demonstrates this invention further more concretely, this invention is not limited to such an Example.
(1) Preparation of support 18 parts by weight of polysulfone ("Udel" P-3500 manufactured by Teijin Amoco), 6 parts by weight of polyvinylpyrrolidone (BASF K90), 3 parts by weight of polyvinylpyrrolidone (BASF K30), N, A solution of 72 parts by weight of N-dimethylacetamide and 1 part by weight of water was prepared. Thereafter, this solution was applied to a glass plate with a 0.1 mm spacer and immediately stretched to form a film with a doctor blade. Immediately after that, the resulting solution was adjusted to 52 parts by weight of N, N-dimethylacetamide and 48 parts by weight of water. The glass plate was immersed to solidify the film. A series of operations were all performed at 50 ° C.
(2) Immobilization of diethylaminoethyl dextran on a support A 1 wt% aqueous solution of diethylaminoethyl dextran (molecular weight of 500,000 manufactured by SIGMA) was prepared. In the state where the polyvinylpyrrolidone blend polysulfone flat membrane prepared in (1) was immersed in an aqueous ethylaminoethyldextran solution, diethylaminoethyldextran was immobilized by irradiating 25 kGy of γ-rays. Thereafter, the mixture was left overnight in 50 ° C. warm water to extract and wash diethylaminoethyl dextran not immobilized on the membrane.
[0030]
The experimental results are shown in Table 1.
Comparative Example 1
A flat polysulfone / polyvinylpyrrolidone flat membrane was prepared in the same manner as in the Examples, filled with water instead of 1 wt% diethylaminoethyldextran aqueous solution, and irradiated with 25 kGy of γ rays.
Comparative Example 2
A polysulfone / polyvinylpyrrolidone flat membrane in which dextran (molecular weight of 500,000 manufactured by SIGMA) was immobilized was prepared in the same manner as in the Examples.
[0031]
Table 1 shows the results of adsorption removal rates of oxidized LDL, LDL, and HDL in Examples and Comparative Examples 1 and 2.
[0032]
[Table 1]

[0033]
【The invention's effect】
According to the present invention, there is provided an adsorbent which is used for applications such as a membrane blood purifier, and which is preferably used in cases where the progression of arteriosclerosis is prevented or prevented in advance in dialysis patients. it can.

Claims (4)

An adsorbent for adsorbing oxidized low density lipoprotein , characterized in that diethylaminoethyldextran is insolubilized by irradiation with a support comprising a blend of polysulfone and polyvinylpyrrolidone. When the support and plasma are allowed to interact with each other under the condition that the plasma volume per 1 m ^{2 of the} support is 3.3 × 10 ² ml / m ² , the initial concentration contained in the plasma is 2 μg / ml. 2. The adsorbent for adsorbing oxidized low density lipoprotein according to claim 1, wherein the adsorption removal rate of oxidized low density lipoprotein is 60% or more. The adsorbent for adsorbing oxidized low density lipoprotein according to claim 1, wherein the adsorption removal rate of low density lipoprotein is less than 20% under the conditions of claim 2. The adsorbent for adsorbing oxidized low-density lipoprotein according to claim 1, wherein the adsorption removal rate of high-density lipoprotein is less than 15% under the conditions of claim 2.