JPH06233815A - Hemocatharsis adsorbent - Google Patents
Hemocatharsis adsorbentInfo
- Publication number
- JPH06233815A JPH06233815A JP50A JP2263993A JPH06233815A JP H06233815 A JPH06233815 A JP H06233815A JP 50 A JP50 A JP 50A JP 2263993 A JP2263993 A JP 2263993A JP H06233815 A JPH06233815 A JP H06233815A
- Authority
- JP
- Japan
- Prior art keywords
- blood
- carrier
- acid
- adsorbent
- polycarboxylic acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
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- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 229940014800 succinic anhydride Drugs 0.000 description 1
- 150000005846 sugar alcohols Polymers 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 229920005613 synthetic organic polymer Polymers 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 1
- 231100000167 toxic agent Toxicity 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 210000003171 tumor-infiltrating lymphocyte Anatomy 0.000 description 1
Landscapes
- External Artificial Organs (AREA)
Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明は自己免疫疾患（重症筋無
力症、慢性リウマチ、特発性血小板減少性紫班病な
ど）、免疫関連疾患（気管支喘息、糸球体腎炎など）、
癌（白血病など）、高脂血症などの治療、及び幹細胞、
Ｂ細胞、Ｔ細胞など血中細胞成分の分離を目的とし、血
液から血漿を分離することなく、直接血液を接触させる
ことによって血中の特定成分を分離する血液浄化吸着材
に関するものである。The present invention relates to autoimmune diseases (myasthenia gravis, chronic rheumatism, idiopathic thrombocytopenic purpura, etc.), immune-related diseases (bronchial asthma, glomerulonephritis, etc.),
Treatment of cancer (leukemia, etc.), hyperlipidemia, and stem cells,
The present invention relates to a blood purification adsorbent for the purpose of separating blood cell components such as B cells and T cells, and separating specific components in blood by directly contacting blood without separating plasma from blood.
【０００２】[0002]
【従来の技術】従来、免疫不全、腫瘍、高脂血症など治
療には血漿交換法が有効とされ、広く行われている。し
かしながら、この療法では血漿をすべて交換するため、
（１）不要成分のみならず、必要な成分までも除去され
てしまうこと、（２）除去した血漿に替えて体内に補充
される血漿、あるいは血漿製剤が不足しているため、大
量かつ持続的な入手が困難である、（３）血清肝炎やア
レルギー、さらにはＡＩＤＳ（後天性免疫不全症候群）
感染の危険性があること、など血漿交換法が内含する問
題は多い。2. Description of the Related Art Conventionally, the plasmapheresis method has been effective and widely used for the treatment of immunodeficiency, tumor, hyperlipidemia and the like. However, this therapy replaces all plasma, so
(1) Not only unnecessary components but also necessary components are removed. (2) A large amount and long-lasting plasma is lacking due to lack of plasma or plasma preparations to replace the removed plasma. Difficult to obtain, (3) Serum hepatitis, allergies, and AIDS (acquired immunodeficiency syndrome)
There are many problems involved in plasmapheresis, such as the risk of infection.
【０００３】こういった状況を背景として、近年、病因
関連物質の選択的除去による療法が盛んに行われるよう
になってきた。このような方法は例えば、メンブレンフ
ィルターを用いるカスケード（Ｓｉｅｂｅｌｔｈ，Ｈ．
Ｇ．，Ｐｌａｓｍａ Ｅｘｃｈａｎｇｅ，Ｐ．２９，
Ｆ．Ｋ．Ｓｃｈａｔｔａｕｅｒ Ｖｅｒｌｏｇ，Ｓｔｕ
ｔｔｇａｒｔ−Ｎｅｗ Ｙｏｒｋ，１９８０）、あるい
は二重ろ過法（阿岸鉄三ら，腎と透析，１０（３），４
７５，１９８１）、凍結ろ過法（Ｌ’Ａｂａｔｔｅ
Ａ．，ｅｔ ａｌ．，Ｐｒｏｃ．Ｅｕｒ．Ｄｉａｌ．Ｔ
ｒａｎｓｐｌａｎｔ．Ａｓｓｏｃ．，１４，４８６，１
９７７）、塩析血漿処理法（大江宏明ら，人工臓器，１
４０（１），４７２−４７５（１９８５））が考案され
ている。Against this background, in recent years, therapy by selective removal of pathogen-related substances has become popular. Such a method is described, for example, in Cascade (Siebelth, H .;
G. , Plasma Exchange, P .; 29,
F. K. Schattauer Verlog, Stu
ttgart-New York, 1980) or double filtration method (Aketsu Tetsuzo et al., Kidney and dialysis, 10 (3), 4).
75, 1981), a freeze filtration method (L'Abatte).
A. , Et al. , Proc. Eur. Dial. T
ranplant. Assoc. , 14 , 486, 1
977), salting-out plasma treatment method (Hiroaki Oe et al., Artificial organ, 1
40 (1), 472-475 (1985)).
【０００４】他の治療方法としては、各種薬剤の使用が
行われているが、一般に副作用の危険性があり、使用
量、使用期間などに細心の注意を払う必要がある。As other treatment methods, various drugs have been used, but generally there is a risk of side effects, and it is necessary to pay close attention to the amount used and the period of use.
【０００５】このような問題を解決できる治療法として
は、自己の血液を浄化した後に再輪注する方法、すなわ
ち、体外循環血液浄化法が望ましいとされる。この方法
を採るにあたり、副作用がなく、自己の血液から病因物
質を充分且つ選択的に除去することのできる浄化材及び
血液浄化療法が望まれていた。As a therapeutic method capable of solving such a problem, a method of purifying own blood and then reinjecting it, that is, an extracorporeal blood purifying method is desirable. In adopting this method, a purification material and a blood purification therapy capable of sufficiently and selectively removing a pathogenic substance from own blood without side effects have been desired.
【０００６】従来、この目的に供し得る血液浄化材とし
ては、 （１）アフィニティー吸着材（例えば、特公平２−５０
９６、特公平２−２６９８８、特開平１−１５８９７０
など） （２）多孔性樹脂（例えば特開昭５６−１４７７１０、
特公昭６２−２５４３、Ｒｏｈｍ＆Ｈａａｓ製の「アン
バーライトＸＡＤ−７」など） （３）イオン交換体（例えばカルボキシメチルセルロー
ス、ジエチルアミノエチルアガロースなど） （４）無機多孔体（例えば特公昭６２−２５４３、多孔
質ガラス、セラミクスなど）がある。Heretofore, as blood purification materials which can be used for this purpose, (1) affinity adsorption materials (for example, Japanese Patent Publication No. 2-50)
96, JP-B-2-26988, JP-A-1-158970.
(2) Porous resin (for example, JP-A-56-147710,
Japanese Patent Publication No. 62-2543, "Amberlite XAD-7" manufactured by Rohm & Haas, etc.) (3) Ion exchangers (for example, carboxymethyl cellulose, diethylaminoethyl agarose, etc.) (4) Inorganic porous substance (for example, Japanese Patent Publication No. 62-2543, porous) Glass, ceramics, etc.).
【０００７】しかし、多孔性樹脂やイオン交換体は吸着
能が小さい上に吸着特異性が低く、これを血液浄化材と
して用いると効果が充分でないばかりでなく、血中の必
須成分までも吸着してしまう可能性があり、安全性の面
でも問題がある。また、無機多孔体は吸着能、吸着特性
については比較的良好であるが、未だ実用に供するには
不十分であり、血液適合性に乏しいため、多量のヘパリ
ンの添加が必要となり、出血傾向等の副作用が生じる。
このような点から、将来的に最も可能性の大きいアフィ
ニティー吸着材による有用な血液浄化材の開発が望まれ
ている。However, porous resins and ion exchangers have low adsorption ability and low adsorption specificity, and when they are used as blood purification materials, not only are the effects insufficient, but they also adsorb essential components in blood. There is a possibility that it will end up, and there is a problem in terms of safety. In addition, although the inorganic porous material is relatively good in adsorption ability and adsorption characteristics, it is still insufficient for practical use and poor in blood compatibility, so it is necessary to add a large amount of heparin, and bleeding tendency, etc. Side effects of.
From these points, it is desired to develop a useful blood purification material using an affinity adsorbent, which has the greatest possibility in the future.
【０００８】アフィニティー吸着材は、生物学アフィニ
ティー吸着材（抗原抗体結合、補体結合、Ｆｃ結合等の
生物学的相互作用で血液中の病因物質と結合し、これを
吸着するもの）と物理化学的アフィニティー吸着材（静
電結合、水素結合、ファンデルワールス力等物理的また
は化学的相互作用によって血液中の病因物質と結合し、
これを吸着するもの）とに大別でき、さらに従来知られ
ている物理化学的アフィニティー吸着材は以下の６つに
分類することができる。 （１）カルボキシル基またはスルホン酸基を表面に有す
る多孔体（例えば、特開昭５６−１４７７１０、特開昭
５７−５６０３８、特開昭５７−７５１４１、特開昭５
７−１７０２６３、特開昭５７−１９７２９４、） （２）疎水性アミノ酸が結合されている親水性担体（例
えば、特開昭５７−１２２８７５、特開昭５８−１５９
２４、特開昭５８−１６５８５９、特開昭５８−１６５
８６１、旭メディカル製イムソーバー） （３）その他疎水性低分子化合物が固定されている担体
（例えば特開平１−１５８９７０） （４）変性免疫グロブリン（ＩｇＧ）が結合されている
親水性担体（例えば特開昭５７−７７６２４、特開昭５
７−７７６２５、特開昭５７−１５６０３５） （５）メチル化アルブミンが結合されている多孔体（例
えば特開昭５５−１２０８７５、特開昭５５−１２５８
７２） （６）糖または改質した糖が結合されている担体（例え
ば特開昭５７−１３４１６４、特開昭５８−１３３２５
７、鐘淵化学リポソーバー） （７）プリン塩基またはピリミジン塩基、或いは糖燐酸
が結合されている多孔体（例えば特開昭５７−１９２５
６０、特開昭５８−６１７５２、特開昭５８−９８１４
２）The affinity adsorbent is a bioaffinity adsorbent (which binds to a pathogenic substance in blood by a biological interaction such as antigen-antibody bond, complement bond, Fc bond, etc. and adsorbs it) and physical chemistry. Affinity adsorbents (electrostatic bond, hydrogen bond, van der Waals force such as physical or chemical interaction with the pathogen in the blood,
This can be roughly classified into the following (6) and the conventionally known physicochemical affinity adsorbents can be classified into the following 6 types. (1) Porous material having a carboxyl group or a sulfonic acid group on its surface (for example, JP-A-56-147710, JP-A-57-56038, JP-A-57-75141, and JP-A-5-75141).
7-170263, JP-A-57-197294, (2) Hydrophilic carrier to which a hydrophobic amino acid is bound (for example, JP-A-57-122875, JP-A-58-159).
24, JP-A-58-165859, JP-A-58-165.
861, Asahi Medical Imsorber) (3) A carrier to which other hydrophobic low molecular weight compounds are fixed (for example, JP-A-1-158970) (4) A hydrophilic carrier to which modified immunoglobulin (IgG) is bound (for example, Kai 57-77624, JP-A-5
7-77625, JP-A-57-156035) (5) Porous substance to which methylated albumin is bound (for example, JP-A-55-10875 and JP-A-55-1258).
72) (6) Carrier to which sugar or modified sugar is bound (for example, JP-A-57-134164 and JP-A-58-13325)
7. Kanegafuchi Chemical Liposorber) (7) Porous body to which purine base or pyrimidine base or sugar phosphate is bound (for example, JP-A-57-1925).
60, JP-A-58-61752, JP-A-58-9814
2)
【０００９】しかし、これら従来の物理化学的アフィニ
ティー吸着材も、体外循環血液浄化療法による自己免疫
疾患等の治療には充分とは言えず、血液適合性にも乏し
いことから、さらに高い効率及び特異性で病因物質を除
去することができ、また体液に対する悪影響の少ない浄
化材が望まれている。However, these conventional physicochemical affinity adsorbents cannot be said to be sufficient for the treatment of autoimmune diseases and the like by extracorporeal blood purification, and are also poor in blood compatibility, and therefore have higher efficiency and specificity. There is a demand for a purification material that can remove the etiological agent and that has less adverse effect on body fluid.
【００１０】このような概念に基づいて開発された血液
浄化材については、例えば、特開平１−１５８９７０と
いう特許が開示されている。この特許によって開示され
ている血液浄化材は、重合度１〜９０のエチレンオキサ
イド骨格を有する親水性スペーサーを介し、被吸着物質
と親和性を有する物質（以下リガンドと略記する）とし
て低分子有機化合物を水不溶性多孔質固体表面に固定
し、これに直接血液を潅流して免疫グロブリンを吸着す
ることを特徴としている。As a blood purification material developed based on such a concept, for example, a patent of Japanese Patent Laid-Open No. 1-158970 is disclosed. The blood purification material disclosed by this patent is a low molecular weight organic compound as a substance (hereinafter abbreviated as a ligand) having an affinity for an adsorbed substance through a hydrophilic spacer having an ethylene oxide skeleton having a polymerization degree of 1 to 90. Is immobilized on the surface of a water-insoluble porous solid, and blood is directly perfused to the surface to adsorb immunoglobulin.
【００１１】親水性スペーサーの持つ意義のひとつとし
て、血液中の血球成分や、被吸着物質でない蛋白の付着
を防止することが挙げられる。親水性スペーサーはその
周辺に水を吸着している。このため吸着材表面は水の層
に覆われ、この水の層が血球成分の付着を防いでいる
（排除体積効果）。また、水の層を形成するスペーサー
が運動することにより、さらに血球成分の付着は困難に
なり、付着した血球成分は脱離しやすくなる。また、凝
固因子や補体の活性化が抑制されるといった利点を有す
る。すなわち、親水性スペーサーの導入によって血液適
合性の向上を図ることができる。One of the meanings of the hydrophilic spacer is to prevent adhesion of blood cell components in blood and proteins that are not adsorbed substances. The hydrophilic spacer adsorbs water around the hydrophilic spacer. Therefore, the surface of the adsorbent is covered with a layer of water, and this layer of water prevents adhesion of blood cell components (excluded volume effect). Further, the movement of the spacer forming the water layer makes it more difficult to attach the blood cell component, and the attached blood cell component is likely to be detached. It also has the advantage that the activation of coagulation factors and complement is suppressed. That is, it is possible to improve blood compatibility by introducing a hydrophilic spacer.
【００１２】しかしながら、血液適合性向上の方法とし
て親水性スペーサーの排除体積効果だけに頼る場合、ス
ペーサーの重合度が９０以下では充分とは言えず、特開
平１−１５８９７０においては血液適合性向上のためア
クリル酸誘導体のコーティングを行っている。このよう
な操作は煩雑であるばかりでなく、毒性をもった物質の
溶出を招く可能性も存在する。However, when relying solely on the excluded volume effect of the hydrophilic spacer as a method for improving blood compatibility, it cannot be said that the degree of polymerization of the spacer is 90 or less, and in JP-A-1-158970, improvement of blood compatibility is not achieved. Therefore, acrylic acid derivative is coated. Such an operation is not only complicated, but may cause the elution of a toxic substance.
【００１３】親水性スペーサーの持つ第二の意義とし
て、リガンドの運動性の上昇によって吸着能の向上が期
待できることが挙げられる。被吸着物質が巨大分子であ
る場合には、リガンドを直接固定しただけでは被吸着物
質とリガンドの接近が困難であり、また一部で結合した
としても結合サイトの数が充分ではなく、容易に脱着し
てしまう可能性が大きい。親水性スペーサーを介在させ
ることによって被吸着物質との接近が促進され、またリ
ガンドの動きに融通性があるため周囲のリガンドが被吸
着物質との結合に参与しやすく、このため結合サイトの
多い強固な結合を生成することができる。The second significance of the hydrophilic spacer is that it is expected that the adsorption ability can be improved by increasing the mobility of the ligand. When the substance to be adsorbed is a macromolecule, it is difficult to approach the substance to be adsorbed and the ligand by directly immobilizing the ligand, and even if some of them are bound, the number of binding sites is not sufficient and it is easy to do so. There is a high possibility that they will be removed. By interposing a hydrophilic spacer, proximity to the substance to be adsorbed is promoted, and since the movement of the ligand is flexible, it is easy for the surrounding ligand to participate in binding with the substance to be adsorbed. Can be created.
【００１４】親水性スペーサーのこのような効果につい
ても、特開平１−１５８９７０に記載されているよう
に、コーティングを行ってしまっては半減するのを否め
ない。With respect to such an effect of the hydrophilic spacer, it is undeniable that the effect will be reduced to half by coating as described in JP-A-1-158970.
【００１５】特公平４−２９３９６では、負電荷を有す
る不溶性担体に被吸着物質と結合可能な官能部位を有す
る有機化合物が結合していることを特徴とする血液浄化
吸着材について開示されている。負電荷を有する表面が
血液凝固や補体活性化を抑制する働きを持っている事実
は１９５１年、Ｓａｗｙｅｒらによって発見され、その
後多くの研究者らによって検討が加えられてきた（例え
ばＳａｗｙｅｒ ｅｔａｌ．，Ａｍｅｒ．Ｊ．Ｓｕｒ
ｇ．，１１４，４２，１９６７、笠井ら．，人工臓器１
２，３２７，１９８３など）。特公平４−２９３９６も
これらの研究と同じく、負電荷による血液適合性の向上
を意図している。Japanese Patent Publication No. 4-29396 discloses a blood purification adsorbent characterized in that an insoluble carrier having a negative charge is bound to an organic compound having a functional site capable of binding to a substance to be adsorbed. The fact that the negatively charged surface has a function of suppressing blood coagulation and complement activation was discovered by Sawyer et al. In 1951, and has been investigated by many researchers thereafter (for example, Sawyer et al. , Amer. J. Sur
g. , 114 , 42, 1967, Kasai et al. , Artificial organ 1
2 , 327, 1983). Japanese Examined Patent Publication (Kokoku) No. 4-29396 also intends to improve blood compatibility by negative charges, as in these studies.
【００１６】しかし、特公平４−２９３９６に開示され
ているように負電荷を有する担体を使用してさえも、特
に繊維径の細い不織布を担体に選び、全血処理を行う場
合には、充分な血球粘着抑制を実現するのは困難であ
る。However, even when a carrier having a negative charge is used as disclosed in Japanese Patent Publication No. 4-29396, it is sufficient when a whole non-woven fabric having a small fiber diameter is selected as a carrier for treating whole blood. It is difficult to achieve effective suppression of blood cell adhesion.
【００１７】[0017]
【発明が解決しようとする課題】本発明は上記従来技術
の欠点を解決し、リガンドの効果を充分に引き出して、
良好な吸着能、吸着特性を有し、コーティング等の煩雑
な操作を省略してなお充分な血液適合性を持った血液浄
化吸着材を提供することにより、副作用がなく、自己の
血液から病因物質を充分且つ選択的に除去し、あるいは
血中の特定成分を選択的に吸着補集することのできる効
果的な体外循環血液浄化療法を実現しようとしたもので
ある。DISCLOSURE OF THE INVENTION The present invention solves the above-mentioned drawbacks of the prior art and brings out the effect of the ligand sufficiently,
By providing a blood purification adsorbent that has good adsorption ability and adsorption characteristics and that does not require complicated operations such as coating and yet has sufficient blood compatibility, there are no side effects and the pathogenic substance from own blood The present invention is intended to realize an effective extracorporeal blood purification treatment capable of sufficiently and selectively removing or selectively adsorbing and collecting a specific component in blood.
【００１８】[0018]
【課題を解決するための手段】本発明の血液浄化吸着材
は、分子量４００〜４００００のポリカルボン酸を導入
した繊維径０．１〜１００μｍの繊維から成る水不溶性
不織布に導入し、両末端にブロモアセチル基を有する親
水性スペーサーを介して、被吸着物質と親和性を有する
有機化合物を固定して成ることを特徴とする。本発明に
使用される担体表面におけるポリカルボン酸由来のカル
ボキシル基含量は、１０〜１０4 μｅｑ／ｇであること
を特徴とする。本発明に使用される親水性スペーサー
は、炭素数２〜６のポリアルキレンオキサイド骨格を有
する繰り返し単位から成り、重合度が２〜４００であっ
て、両末端にブロモアセチル基を有する化１に示した構
造であることを特徴とする。The blood purification adsorbent of the present invention is introduced into a water-insoluble non-woven fabric composed of fibers having a fiber diameter of 0.1 to 100 μm and containing a polycarboxylic acid having a molecular weight of 400 to 40,000, and is applied to both ends. It is characterized in that an organic compound having an affinity with the substance to be adsorbed is immobilized via a hydrophilic spacer having a bromoacetyl group. The content of the carboxyl group derived from the polycarboxylic acid on the surface of the carrier used in the present invention is characterized by being 10 to 10 4 μeq / g. The hydrophilic spacer used in the present invention is composed of a repeating unit having a polyalkylene oxide skeleton having 2 to 6 carbon atoms, has a degree of polymerization of 2 to 400, and has a bromoacetyl group at both ends. It has a different structure.
【００１９】本発明の不織布型血液浄化吸着材に用いら
れる水不溶性不織布は、ポリエステル、ポリエチレン、
ポリプロピレン、ポリスチレン、ポリメタクリル酸およ
びその誘導体或いはこれらの共重合体などの合成有機高
分子化合物、セルロース、セファロース、デキストラ
ン、キチン、キトサン等の天然有機高分子化合物、また
はアシルセルロース、アシルセファロース等の改質天然
有機高分子化合物であることが好ましい。これらの高分
子化合物のうち、機械的強度、改質の容易さ等を考慮す
ると、ポリエステル、ポリエチレン、ポリスチレンおよ
びその誘導体或いはこれらの共重合体、エチレン−ビニ
ルアルコール共重合体、あるいはセルロース、キトサン
が望ましく、さらに好ましくはポリエステル、ポリエチ
レン、セルロースが望ましい。The water-insoluble nonwoven fabric used in the nonwoven fabric-type blood purification adsorbent of the present invention includes polyester, polyethylene,
Synthetic organic polymer compounds such as polypropylene, polystyrene, polymethacrylic acid and its derivatives or copolymers thereof, natural organic polymer compounds such as cellulose, sepharose, dextran, chitin and chitosan, or modified acylcellulose, acyl sepharose and the like. It is preferably a natural organic polymer compound. Among these polymer compounds, considering mechanical strength, easiness of modification, etc., polyester, polyethylene, polystyrene and derivatives thereof or copolymers thereof, ethylene-vinyl alcohol copolymer, or cellulose, chitosan Desirably, more preferably polyester, polyethylene, or cellulose.
【００２０】本発明は担体として水不溶性不織布を使用
することが特徴のひとつとなっている。本発明において
は血液から血漿を分離することなく直接全血を潅流して
血中の特定成分を分離することを目的としているため、
吸着材と血液が接触する際の流通抵抗をいかに低く抑え
るかが大きな問題となる。先願特許の多くは担体の形状
として、粒子状、繊維状、中空糸状、膜状などさまざま
なものを例示してはいるものの、実質的には粒子状担体
を使用することを前提としていることが多い。しかしな
がら、粒子状担体では有効表面積を獲得するのは比較的
容易だが、流通抵抗を低く抑えるには必ずしも適当であ
るとは言い難い。このように圧損と有効表面積の両立を
考えた場合、繊維状担体を使用することが好ましく、さ
らにその繊維状担体は不織布になっていることが好まし
いとの結論に至った。One of the features of the present invention is the use of a water-insoluble nonwoven fabric as a carrier. In the present invention, since it is intended to directly perfuse whole blood without separating plasma from blood to separate a specific component in blood,
A major problem is how to reduce the flow resistance when the adsorbent comes into contact with blood. Although many of the prior patents exemplify various shapes such as particles, fibers, hollow fibers, and membranes as the shape of the carrier, it is premised that the particulate carrier is substantially used. There are many. However, although it is relatively easy to obtain an effective surface area with a particulate carrier, it is difficult to say that it is necessarily suitable for keeping the flow resistance low. In this way, when considering both the pressure loss and the effective surface area, it was concluded that it is preferable to use a fibrous carrier, and it is preferable that the fibrous carrier is a non-woven fabric.
【００２１】不織布型担体を形成する繊維の径は０．１
〜１００μｍの範囲にあることが必要であるが、好まし
くは１〜５０μｍ、さらに好ましくは３〜２０μｍであ
ることが好ましい。これよりも繊維径が小さくなると血
球の流通抵抗が大きくなり、繊維径が大きくなると有効
表面積の減少を招く。繊維の形状については、細胞に与
える損傷などから考えて、刺状の突起が存在することは
好ましくないが、有効表面積を増加させる目的で微小孔
が存在することは好ましい。その平均孔径は２０Å〜３
０００Å、好ましくは１００Å〜２０００Åである。こ
れより平均孔径が小さいと表面積の実測値は大きくなる
ものの、被吸着物質が微小孔内部に到達できないため有
効表面積の拡大にはならず、これより大きいと担体の強
度低下を招く恐れがある。また、繊維にクリンプが存在
することも、流通抵抗の低下や血液との接触効率の向上
につながるため好ましい。本発明の不織布は０．６ｇ／
ｃｍ3 以下、好ましくは０．４ｇ／ｃｍ3 以下のもので
あり、この下限は０．０１ｇ／ｃｍ3 のものが好まし
い。The diameter of the fibers forming the non-woven fabric type carrier is 0.1
It is necessary to be in the range of -100 μm, preferably 1-50 μm, more preferably 3-20 μm. When the fiber diameter is smaller than this, the circulation resistance of blood cells increases, and when the fiber diameter increases, the effective surface area decreases. Regarding the shape of the fiber, it is not preferable that spiny protrusions are present in view of damage to cells, but it is preferable that micropores are present for the purpose of increasing the effective surface area. The average pore size is 20Å ~ 3
It is 000Å, preferably 100Å to 2000Å. If the average pore diameter is smaller than this, the measured value of the surface area will be large, but since the substance to be adsorbed cannot reach the inside of the micropores, the effective surface area will not be expanded, and if it is larger than this, the strength of the carrier may be reduced. The presence of crimps in the fibers is also preferable because it leads to a reduction in flow resistance and an improvement in contact efficiency with blood. The nonwoven fabric of the present invention is 0.6 g /
cm 3 or less, preferably 0.4 g / cm 3 or less, and the lower limit is preferably 0.01 g / cm 3 .
【００２２】本発明に用いられる分子量４００〜４００
００のポリカルボン酸としては、アクリル酸、ビニル安
息香酸、メタクリル酸、マレイン酸、フマル酸などのカ
ルボキシル基含有ビニル系化合物の重合体、あるいはカ
ルボキシル基含有ビニル系化合物を共重合成分とする共
重合体；ポリグルタミン酸、ポリアスパラギン酸などの
酸性ポリアミノ酸、あるいはこれらを共重合成分とする
酸性共重合体；アルギン酸、ペクチン酸などの酸性多糖
類などが挙げられる。また、ポリメタクリル酸メチル、
ポリアクリルアミド、ポリアクリロニトリル、ポリ無水
マレイン酸の加水分解物なども使用され得る。これらの
ポリカルボン酸のうち、導入の容易さや導入後の安定
性、親水性などから、アクリル酸、ビニル安息香酸、メ
タクリル酸、マレイン酸、フマル酸などのカルボキシル
基含有ビニル系化合物重合体が好ましく、さらに好まし
くはポリアクリル酸、ポリメタクリル酸が好ましい。Molecular weight used in the present invention 400-400
The polycarboxylic acid of 00 is a polymer of a carboxyl group-containing vinyl compound such as acrylic acid, vinyl benzoic acid, methacrylic acid, maleic acid or fumaric acid, or a copolymer containing a carboxyl group-containing vinyl compound as a copolymerization component. Examples of the compound include acidic polyamino acids such as polyglutamic acid and polyaspartic acid, and acidic copolymers containing these as copolymerization components; acidic polysaccharides such as alginic acid and pectic acid. Also, polymethylmethacrylate,
Polyacrylamide, polyacrylonitrile, hydrolysates of polymaleic anhydride, etc. may also be used. Of these polycarboxylic acids, carboxyl group-containing vinyl compound polymers such as acrylic acid, vinyl benzoic acid, methacrylic acid, maleic acid and fumaric acid are preferred from the viewpoint of ease of introduction and stability after introduction, hydrophilicity and the like. , And more preferably polyacrylic acid and polymethacrylic acid.
【００２３】本発明においてポリカルボン酸を担体に導
入し、リガンドの固定化を行う意義は大きくわけて二つ
挙げられる。第一は担体表面に負電荷を導入することで
血液凝固や補体活性化を抑制し、血液適合性を向上させ
ることである。血球は負に帯電しているため、負電荷を
有する表面とは静電反発によって吸着が阻害され、凝血
が起こりにくくなる。また、補体活性化のカスケードが
負電荷素材のトラップによって阻害を受けるため、補体
活性化抑制の効果も期待できる。In the present invention, the significance of immobilizing a ligand by introducing a polycarboxylic acid into a carrier can be broadly divided into two. The first is to suppress blood coagulation and complement activation by introducing a negative charge on the surface of the carrier to improve blood compatibility. Since the blood cells are negatively charged, electrostatic repulsion from the surface having a negative charge inhibits adsorption, and coagulation is less likely to occur. Moreover, since the cascade of complement activation is inhibited by the trap of the negatively charged material, an effect of suppressing complement activation can be expected.
【００２４】負電荷を付与するにはカルボキシル基のほ
かにスルホン酸基、リン酸基なども考えられるが、これ
ら種々の負電荷を有する置換基について血液適合性を検
討した結果、特に血球粘着についてはカルボキシル基を
導入するのが最も好ましい結果となった。To impart a negative charge, a sulfonic acid group, a phosphoric acid group, etc. can be considered in addition to the carboxyl group. As a result of examining the blood compatibility of these various negatively charged substituents, especially regarding blood cell adhesion. The most preferable result was to introduce a carboxyl group.
【００２５】また、低分子量のカルボキシル基含有化合
物で担体表面に負電荷を導入した場合と比較して、分子
量が４００〜４００００の範囲のポリカルボン酸を水不
溶性担体に導入した場合には、より良好な血液適合性、
吸着性能を得ることができる。この理由として以下のよ
うなことが考えられる。すなわち、血液などの液体中で
は、ポリカルボン酸の分子鎖の一部が担体表面から立ち
上がって表面近傍に揺らぐように存在する可能性が期待
できる。この立ち上がったポリカルボン酸分子鎖は負電
荷を有し、親水性であるため排除体積効果が期待でき、
血液適合性の向上に寄与すると同時に、リガンドの運動
性を上昇させることで吸着能の向上をももたらす。つま
り、ポリカルボン酸の第二の効果として、親水性スペー
サーとしての働きを期待できる。Further, compared with the case where a negative charge is introduced on the carrier surface with a low molecular weight compound containing a carboxyl group, when a polycarboxylic acid having a molecular weight in the range of 400 to 40,000 is introduced into a water-insoluble carrier, Good blood compatibility,
Adsorption performance can be obtained. The reasons for this are as follows. That is, in a liquid such as blood, it can be expected that a part of the molecular chain of polycarboxylic acid exists so as to rise from the surface of the carrier and swing near the surface. Since this raised polycarboxylic acid molecular chain has a negative charge and is hydrophilic, an excluded volume effect can be expected,
At the same time as contributing to the improvement of blood compatibility, it also improves the adsorption ability by increasing the mobility of the ligand. That is, the second effect of the polycarboxylic acid can be expected to act as a hydrophilic spacer.
【００２６】単にカルボキシル基含有低分子量化合物の
固定化や、負電荷を有する素材を担体に使用するだけで
は上記のような排除体積効果は期待できず、充分な血液
適合性を得るのは困難であり、リガンドの効果も充分に
引き出されない。すなわち、良好な血液適合性や吸着性
能を得るには、担体表面に負電荷を付与するのにあたり
分子量４００〜４００００のポリカルボン酸を用いるこ
とが不可欠である。このような観点から、より好ましい
ポリカルボン酸の分子量は５００〜２００００であり、
さらに好ましくは１０００〜１００００であることが好
ましい。これよりも分子量が小さいと充分な排除体積効
果、リガンドの運動性は得られず、これ以上の分子量だ
と導入効率が低下するため好ましくない。The above-mentioned excluded volume effect cannot be expected simply by immobilizing a carboxyl group-containing low molecular weight compound or using a material having a negative charge as a carrier, and it is difficult to obtain sufficient blood compatibility. Yes, the effect of the ligand is not fully elicited. That is, in order to obtain good blood compatibility and adsorption performance, it is essential to use a polycarboxylic acid having a molecular weight of 400 to 40,000 to impart a negative charge to the carrier surface. From this point of view, the more preferable molecular weight of the polycarboxylic acid is 500 to 20,000,
It is more preferably 1000 to 10,000. If the molecular weight is smaller than this, sufficient excluded volume effect and ligand mobility cannot be obtained, and if the molecular weight is higher than this, the introduction efficiency decreases, which is not preferable.
【００２７】しかしながら、ポリカルボン酸だけでは親
水性スペーサーとしての効果はいまだ充分であるとは言
い難く、より良好な吸着性能、血液適合性を得るために
は非イオン性の親水性スペーサーが必要である。However, it cannot be said that the effect as a hydrophilic spacer is still sufficient with polycarboxylic acid alone, and a nonionic hydrophilic spacer is necessary to obtain better adsorption performance and blood compatibility. is there.
【００２８】本発明に使用される親水性スペーサーは化
１に示された構造を有することが特徴である。その基本
骨格としては、ポリエチレングリコール、ポリプロピレ
ングリコール、ポリテトラメチレングリコールなどが例
示されるが、構造が化１に示した一般式に合致するかぎ
り、これらに限定されるものではない。The hydrophilic spacer used in the present invention is characterized by having the structure shown in Chemical formula 1. Examples of the basic skeleton include polyethylene glycol, polypropylene glycol, polytetramethylene glycol and the like, but the basic skeleton is not limited to these as long as the structure conforms to the general formula shown in Chemical formula 1.
【００２９】ポリカルボン酸を導入した不織布型担体
に、両末端にブロモアセチル基を有する親水性スペーサ
ーを介してリガンドを固定するのが本発明の要旨である
が、担体にポリカルボン酸を導入した後親水性スペーサ
ーを導入し、さらにリガンドを固定する方法；担体にポ
リカルボン酸を導入した後、あらかじめリガンドを固定
した親水性スペーサーを導入する方法；親水性スペーサ
ーを固定したポリカルボン酸を担体に導入した後リガン
ドを固定する方法；親水性スペーサーを介してリガンド
を固定したポリカルボン酸を担体に導入する方法、いず
れを採用してもよく、最終的に得られる血液浄化吸着材
に存在するポリカルボン酸由来のカルボキシル基含量が
１０〜１０4 μｅｑ／ｇであり、両末端にブロモアセチ
ル基を有する親水性スペーサーを介してリガンドが固定
されている限りその製造方法は制限を受けるものではな
い。It is the gist of the present invention to immobilize a ligand on a non-woven fabric type carrier into which a polycarboxylic acid has been introduced via hydrophilic spacers having bromoacetyl groups at both ends. The polycarboxylic acid was introduced into the carrier. A method of introducing a hydrophilic spacer afterwards and further immobilizing a ligand; a method of introducing a polycarboxylic acid onto a carrier and then introducing a hydrophilic spacer onto which a ligand is previously immobilized; a polycarboxylic acid onto which a hydrophilic spacer is immobilized onto a carrier Any method may be employed, in which the ligand is immobilized after the introduction; the method in which the ligand-immobilized polycarboxylic acid is introduced into the carrier through a hydrophilic spacer, any of which is present in the finally obtained blood purification adsorbent. carboxyl group content derived from a carboxylic acid is 10~10 4 μeq / g, hydrophilicity with bromoacetyl groups on both ends Its production method as long as the ligand through the pacer is fixed not to be limited.
【００３０】さらには、担体表面に導入したポリカルボ
ン酸とは別個に、独立して親水性スペーサーを担体表面
に導入し、この親水性スペーサーの片末端にリガンドを
固定化するという方法も採用され得る。この場合、ポリ
カルボン酸の分子鎖は負電荷の付与と排除体積効果によ
って担体の血液適合性向上に貢献し、担体表面に導入さ
れた親水性スペーサーは排除体積効果によって血液適合
性向上に貢献するとともに、リガンドの運動性向上に寄
与する。つまり、ポリカルボン酸分子鎖と親水性スペー
サーによって血液適合性向上とリガンドの運動性向上と
が、ある程度役割分担されることになる。Furthermore, a method of independently introducing a hydrophilic spacer to the surface of the carrier separately from the polycarboxylic acid introduced to the surface of the carrier and immobilizing a ligand on one end of this hydrophilic spacer is also adopted. obtain. In this case, the molecular chain of polycarboxylic acid contributes to the improvement of blood compatibility of the carrier by imparting a negative charge and the excluded volume effect, and the hydrophilic spacer introduced on the surface of the carrier contributes to the improvement of blood compatibility by the excluded volume effect. At the same time, it contributes to improving the mobility of the ligand. In other words, the polycarboxylic acid molecular chain and the hydrophilic spacer are responsible for improving blood compatibility and ligand mobility to some extent.
【００３１】本発明に使用される水不溶性不織布型担体
へのポリカルボン酸は、使用中に遊離が認められない限
り特に制限されるものではないが、化学反応による共有
結合、イオン結合、電子線や紫外線の照射によるグラフ
ト化などが好ましい。特に、不織布型担体の素材となる
のがセルロースやキトサンなどの場合には化学反応によ
る共有結合での固定化が好ましく、素材がポリエチレン
やポリエステルなどの場合には電子線照射によるグラフ
ト化が好ましい。The polycarboxylic acid for the water-insoluble non-woven fabric type carrier used in the present invention is not particularly limited as long as it is not liberated during use, but covalent bond by a chemical reaction, ionic bond, electron beam. Grafting by irradiation with or ultraviolet rays is preferable. Particularly, when the material of the non-woven fabric type carrier is cellulose, chitosan or the like, immobilization by a covalent bond by a chemical reaction is preferable, and when the material is polyethylene, polyester or the like, grafting by electron beam irradiation is preferable.
【００３２】セルロース製の担体には、過沃素酸塩との
反応で導入したアルデヒド基、モノクロル酢酸との反応
で導入したカルボキシル基、無水コハク酸との反応で導
入したカルボキシル基、塩化パラトルエンスルホニルと
の反応で導入した活性エステルなどの活性置換基を足が
かりとして利用し、数ステップの反応を経ることによっ
て化学結合でポリカルボン酸を固定化することができ
る。The cellulose carrier includes an aldehyde group introduced by a reaction with periodate, a carboxyl group introduced by a reaction with monochloroacetic acid, a carboxyl group introduced by a reaction with succinic anhydride, and paratoluenesulfonyl chloride. By using an active substituent such as an active ester introduced by the reaction with and as a foothold, the polycarboxylic acid can be immobilized by a chemical bond through a reaction of several steps.
【００３３】不織布型担体に電子線照射でポリアクリル
酸を導入する場合、ポリアクリル酸を溶解することがで
き、揮発性である溶媒にポリアクリル酸を溶解し、この
溶液を不織布型担体に塗布して乾燥した後、電子線を照
射する方法が好ましい。揮発性の溶媒としては特に限定
されるものではないが、水、メタノール、エタノール、
塩化メチレン、クロロホルム、アセトン、ジオキサン、
テトラヒドロフラン、またはこれらの混合溶媒が好まし
い。When polyacrylic acid is introduced into the non-woven fabric carrier by electron beam irradiation, polyacrylic acid can be dissolved, and the polyacrylic acid is dissolved in a volatile solvent, and this solution is applied to the non-woven fabric carrier. The method of irradiating with an electron beam after drying is preferable. The volatile solvent is not particularly limited, but water, methanol, ethanol,
Methylene chloride, chloroform, acetone, dioxane,
Tetrahydrofuran or a mixed solvent thereof is preferable.
【００３４】しかし、この方法を用いてもポリカルボン
酸の沸点が低い場合には、溶媒の乾燥工程中にポリカル
ボン酸の一部または全部が蒸発してしまい、グラフト量
の制御が困難になる可能性がある。また、逆にポリカル
ボン酸の融点が高い場合、溶媒の乾燥後、ポリカルボン
酸が固体として析出してしまうため不織布型担体との接
触効率が低下してしまい、グラフト量が低下する可能性
がある。However, even if this method is used, if the boiling point of the polycarboxylic acid is low, a part or all of the polycarboxylic acid will evaporate during the solvent drying step, making it difficult to control the graft amount. there is a possibility. On the other hand, if the melting point of the polycarboxylic acid is high, the polycarboxylic acid will precipitate as a solid after the solvent is dried, so that the contact efficiency with the non-woven fabric type carrier will decrease, and the graft amount may decrease. is there.
【００３５】このような事態を回避して電子線照射によ
るポリカルボン酸のグラフト効率を向上させるには、塗
布溶液中にポリカルボン酸を溶解できる高沸点の化合物
を添加することが好ましい。このような高沸点化合物と
しては、特に限定されるものではないが、エチレングリ
コール、プロピレングリコール、グリセリン等の多価ア
ルコール、およびこれらの脱水縮合物のうち重合度が１
０以下で、流動性を持っているものなどが好ましい。こ
れら高沸点添加物の添加量はポリカルボン酸の重量に対
して、１／１０〜１０／１程度が好ましい。In order to avoid such a situation and improve the grafting efficiency of the polycarboxylic acid by electron beam irradiation, it is preferable to add a high boiling point compound capable of dissolving the polycarboxylic acid in the coating solution. Such high boiling point compounds are not particularly limited, but polyhydric alcohols such as ethylene glycol, propylene glycol, glycerin, and dehydration condensates thereof having a degree of polymerization of 1
Those having a fluidity of 0 or less are preferable. The addition amount of these high boiling point additives is preferably about 1/10 to 10/1 with respect to the weight of the polycarboxylic acid.
【００３６】高沸点化合物よりもさらに効果的にポリカ
ルボン酸の導入を促進する添加物として、架橋性ビニル
化合物があげられる。この架橋性ビニル化合物として
は、メチレンビスアクリルアミド、トリメチロールプロ
パンジアクリレート、トリアリルイソシアヌレート、ト
リメチロールプロパントリアクリレート、テトラメチロ
ールメタンテトラアクリレートなどが例示され、より好
ましくは下記化２に示したような化合物が好ましい。化
２に示した構造を有する添加物の添加量はポリカルボン
酸の重量に対して１／５０〜５０／１、好ましくは１／
３０〜３０／１である。A crosslinkable vinyl compound is an additive that promotes the introduction of the polycarboxylic acid more effectively than the high boiling point compound. Examples of the crosslinkable vinyl compound include methylenebisacrylamide, trimethylolpropane diacrylate, triallyl isocyanurate, trimethylolpropane triacrylate, tetramethylolmethane tetraacrylate, and the like, more preferably as shown in Chemical formula 2 below. Compounds are preferred. The amount of the additive having the structure shown in Chemical formula 1 is 1/50 to 50/1, preferably 1/50 to the weight of the polycarboxylic acid.
It is 30 to 30/1.
【００３７】化２において、ｐ＝１〜３０の整数 Ｒ3 、Ｒ4 、Ｒ5 は水素原子またはメチル基で、それぞ
れ同じもしくは異なってもよい。In the chemical formula 2, the integers R 3 , R 4 and R 5 of p = 1 to 30 are hydrogen atoms or methyl groups and may be the same or different.
【化２】 [Chemical 2]
【００３８】本発明に使用される水不溶性不織布型担体
への、両末端にブロモアセチル基を有する親水性スペー
サーの導入方法は共有結合、電子線照射によるグラフト
化など、使用中に遊離が認められない限り特に制限され
るものではないが、ブロモアセチル基の反応性を利用し
て共有結合によって固定するのが最も好ましい。The method for introducing the hydrophilic spacer having bromoacetyl groups at both ends into the water-insoluble non-woven fabric carrier used in the present invention is covalent bond, grafting by electron beam irradiation, etc., and release is observed during use. It is not particularly limited as long as it is not fixed, but it is most preferable to fix it by a covalent bond by utilizing the reactivity of the bromoacetyl group.
【００３９】ブロモアセチル基はアミノ基、イミノ基、
水酸基、チオール基などと反応することが知られてい
る。この反応を利用して固定化を行うことが好ましい。
アミノ基とブロモアセチル基の反応について下記化３に
示した。この反応による固定化法が比較的簡便で、生成
物も比較的安定であり、推奨される。The bromoacetyl group is an amino group, an imino group,
It is known to react with hydroxyl groups and thiol groups. Immobilization is preferably performed by utilizing this reaction.
The reaction between the amino group and the bromoacetyl group is shown in Chemical Formula 3 below. The immobilization method by this reaction is relatively simple and the product is also relatively stable, which is recommended.
【００４０】[0040]
【化３】 [Chemical 3]
【００４１】化３に示した通り、ブロモアセチル基の反
応性を利用して固定化反応を行った場合、ブロモアセチ
ル基の構造は改変される。従って本発明において、両末
端にブロモアセチル基を有する親水性スペーサーを介し
て固定する、とはこのようにブロモアセチル基の反応性
を利用して固定化反応を行った結果、該親水性スペーサ
ーの構造の一部が改変される場合をも包含する。As shown in Chemical formula 3, when the immobilization reaction is carried out by utilizing the reactivity of the bromoacetyl group, the structure of the bromoacetyl group is modified. Therefore, in the present invention, the immobilization via hydrophilic spacers having bromoacetyl groups at both ends means that the immobilization reaction is performed by utilizing the reactivity of the bromoacetyl group as a result, It also includes the case where a part of the structure is modified.
【００４２】本発明の吸着材が対象とする被吸着物質は
血液中の種々の特定物質であるが、より詳細には通常の
免疫グロブリン（Ａ、Ｄ、Ｅ、Ｇ、Ｍ）、抗ＤＮＡ抗
体、抗アセチルコリンレセプター抗体、抗血液型抗体、
抗血小板抗体などの自己抗体；抗原・抗体複合物；エン
ドトキシン；リウマチ因子；ＬＤＬ、ＶＬＤＬなどのリ
ポ蛋白；幹細胞、Ｂ細胞、Ｔ細胞、単球、マクロファー
ジ、ＴＩＬ（癌組織浸潤Ｔ細胞）などの血中細胞成分な
どが挙げられる。The substances to be adsorbed targeted by the adsorbent of the present invention are various specific substances in blood. More specifically, ordinary immunoglobulins (A, D, E, G, M) and anti-DNA antibodies are used. , Anti-acetylcholine receptor antibody, anti-blood group antibody,
Autoantibodies such as antiplatelet antibodies; antigen-antibody complexes; endotoxins; rheumatoid factors; lipoproteins such as LDL and VLDL; stem cells, B cells, T cells, monocytes, macrophages, TILs (cancer tissue infiltrating T cells), etc. Examples include cell components in blood.
【００４３】固定するリガンドは被吸着物質によって適
宜変える必要があるが、抗原抗体結合、補体結合、Ｆｃ
結合等の生物学的相互作用を利用する種々の生物学的リ
ガンド；または、疎水性アミノ酸、メチル化アルブミ
ン、変性免疫グロブリン（ＩｇＧ）、糖または改質した
糖、プリン塩基、ピリミジン塩基、糖燐酸などの、静電
結合、水素結合、ファンデルワールス力等物理的または
化学的相互作用を利用した種々の物理化学的リガンドが
考えられる。The ligand to be immobilized needs to be appropriately changed depending on the substance to be adsorbed, but it may be antigen-antibody binding, complement binding, Fc
Various biological ligands that utilize biological interactions such as binding; or hydrophobic amino acids, methylated albumin, modified immunoglobulins (IgG), sugars or modified sugars, purine bases, pyrimidine bases, sugar phosphates Various physicochemical ligands utilizing physical or chemical interactions such as electrostatic bond, hydrogen bond, van der Waals force, etc. are considered.
【００４４】リガンドの固定方法は、共有結合、物理的
吸着、イオン結合、生化学的特異結合等特に制限されな
いが、血液中での結合安定性を考慮すると、ブロモアセ
チル基の反応性を利用した共有結合が好ましい。すなわ
ち、リガンドに含まれるアミノ基、イミノ基、チオール
基などとブロモアセチル基の反応を利用し、固定化する
方法が推奨される。The method for immobilizing the ligand is not particularly limited, such as covalent bond, physical adsorption, ionic bond, biochemical specific bond, etc. However, considering the bond stability in blood, the reactivity of the bromoacetyl group was used. Covalent bonds are preferred. That is, a method of immobilizing by using the reaction of an amino group, an imino group, a thiol group and the like contained in the ligand with a bromoacetyl group is recommended.
【００４５】本発明の血液浄化吸着材は、体液の導出入
口を備えた適当な容器に充填することによって使用され
得る。容器の材質はガラス、ステンレス、ポリエチレ
ン、ポリプロピレン、ポリカーボネート、ポリスチレ
ン、ポリメチルメタクリレート等、特に制限されない
が、滅菌等の取扱いを考慮すると、ポリプロピレンやポ
リカーボネートが好ましい。容器の形態についても特に
制限されないが、両端を血液流入部、血液流出部とした
円筒のカラム型、或いはシート状に成形した後、これを
挟み込むような形態にしたものが適当であろう。The blood purification adsorbent of the present invention can be used by filling it in a suitable container having a body fluid outlet port. The material of the container is not particularly limited, such as glass, stainless steel, polyethylene, polypropylene, polycarbonate, polystyrene, polymethylmethacrylate, etc., but polypropylene and polycarbonate are preferable in consideration of handling such as sterilization. The form of the container is also not particularly limited, but a cylindrical column type with both ends having a blood inflow part and a blood outflow part, or a sheet-like shape and then sandwiching the container may be suitable.
【００４６】本発明の血液浄化吸着材は単独で使用して
もよく、活性炭や多孔質ガラス、シリカゲルなど他の吸
着材と混合、積層してもよい。また、リガンドとして２
種以上の有機化合物を固定すること、異なったリガンド
を固定化した吸着材を混合、積層して用いることも制限
を受けるものではない。The blood purification adsorbent of the present invention may be used alone, or may be mixed and laminated with another adsorbent such as activated carbon, porous glass or silica gel. Also, 2 as a ligand
There is no limitation on immobilizing one or more kinds of organic compounds and mixing and laminating adsorbents having different ligands immobilized thereon.
【００４７】[0047]
【実施例】以下、実施例を用いて本発明を説明する。 〈実施例１〉 （１）担体の合成 繊維径３．５μｍ、目付け４５ｇ／ｍ2 のポリエチレン
テレフタレート（以下ＰＥＴと略記する）製の不織布を
１５ｃｍ×１２ｃｍの大きさに切断した（重量約８１０
ｍｇ）。このＰＥＴ製不織布は改質に先立ち、アセトン
で充分洗浄しておいた。分子量４０００のポリアクリル
酸（以下ＰＡＡと略記する）１５ｇ、メチレンビスアク
リルアミド（以下ＭＢＡＡと略記する）５ｇ、グリセリ
ン２ｇをエタノール２ｌに溶解し、この溶液にＰＥＴ製
不織布を浸漬して、ＰＡＡの塗布を行った。これを充分
に乾燥させた後、片面につき５Ｍｒａｄの線量で電子線
（以下ＥＢと略記する）を照射し、ＰＥＴ表面へのＰＡ
Ａのグラフト化を行った。水、メタノールで充分に洗浄
し、ＰＡＡ導入ＰＥＴ不織布（ＰＥＴ−ＰＡＡ）を得
た。得られたＰＥＴ−ＰＡＡは１２０枚であった。この
ＰＥＴ−ＰＡＡの酸含量を平沼産業製ＣＯＭＴＩＴＥ１
０１によって酸塩基滴定で定量したところ、０．３２ｍ
ｅｑ／ｇであった。EXAMPLES The present invention will be described below with reference to examples. Example 1 (1) Synthesis of Carrier A nonwoven fabric made of polyethylene terephthalate (hereinafter abbreviated as PET) having a fiber diameter of 3.5 μm and a basis weight of 45 g / m 2 was cut into a size of 15 cm × 12 cm (weight: about 810).
mg). This PET non-woven fabric was thoroughly washed with acetone before modification. 15 g of polyacrylic acid having a molecular weight of 4000 (hereinafter abbreviated as PAA), 5 g of methylenebisacrylamide (hereinafter abbreviated as MBAA), and 2 g of glycerin are dissolved in 2 l of ethanol, and a PET non-woven fabric is dipped in this solution to apply PAA. I went. After drying it sufficiently, it was irradiated with an electron beam (hereinafter abbreviated as EB) at a dose of 5 Mrad on one side to PA on the PET surface.
Grafting of A was performed. It was thoroughly washed with water and methanol to obtain a PAA-introduced PET nonwoven fabric (PET-PAA). The obtained PET-PAA was 120 sheets. The acid content of this PET-PAA is COMMITTE1 made by Hiranuma Sangyo.
As determined by acid-base titration with 01, 0.32 m
It was eq / g.
【００４８】（２）スペーサー固定用アミノ基の導入 ガラス瓶に１−エチル−３−（３−ジメチルアミノプロ
ピル）−カルボジイミド塩酸塩（以下ＥＤＣと略記す
る）２．４ｇを取り、ｐＨ４．５のリン酸緩衝液５００
ｍｌを加えて溶解させた。このガラス瓶にジエチレント
リアミン３．２ｇを加えてＨＣｌでｐＨを調節し、これ
に、上記で得たＰＥＴ−ＰＡＡ４０枚を浸漬して室温で
約１８時間振盪して反応させた。生成物を取り出し、充
分に洗浄して、アミノ基導入ＰＥＴ−ＰＡＡ（ＰＥＴ−
ＰＡＡ／ＮＨ2）を得た。酸塩基滴定で残存しているカ
ルボキシル基を定量したところ、０．２０ｍｅｑ／ｇで
あった。また、導入されたアミノ基含量は０．１１ｍｅ
ｑ／ｇであった。(2) Introduction of spacer-immobilizing amino group 2.4 g of 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride (hereinafter abbreviated as EDC) was placed in a glass bottle, and phosphorus having a pH of 4.5 was used. Acid buffer 500
ml was added and dissolved. To this glass bottle, 3.2 g of diethylenetriamine was added, and the pH was adjusted with HCl. 40 pieces of the PET-PAA obtained above were dipped in this and shaken at room temperature for about 18 hours to react. The product is taken out, washed thoroughly, and amino group-introduced PET-PAA (PET-PAA
PAA / NH 2 ) was obtained. The residual carboxyl group was determined by acid-base titration to be 0.20 meq / g. In addition, the introduced amino group content is 0.11 me
It was q / g.
【００４９】（３）末端ブロモアセチル基含有スペーサ
ーの導入 ガラス瓶に上記の操作で得たＰＥＴ−ＰＡＡ／ＮＨ2 ４
０枚（約３２．５ｇ）を取った。両末端にブロモアセチ
ル基を有するポリエチレングリコール（ポリエチレング
リコールの分子量は２０００、以下この化合物をＰＥＢ
Ａと略記する）１５ｇをｐＨ８．５のリン酸緩衝液６５
０ｍｌに溶解した。この溶液をＰＥＴ−ＰＡＡ／ＮＨ2
の入ったガラス瓶に注ぎ、ＰＥＴ−ＰＡＡ／ＮＨ2 が浸
るようにした。２５℃で約１４時間振盪して反応させた
後、生成物を取り出し、イオン交換水で充分に洗浄し
た。こうしてＰＥＡを固定化したＰＥＴ−ＰＡＡ／ＮＨ
2 （ＰＥＴ−ＰＡＡ−ＰＥＢＡ）を得た。(3) Introduction of terminal bromoacetyl group-containing spacer PET-PAA / NH 2 4 obtained by the above operation in a glass bottle.
0 sheets (about 32.5 g) were taken. Polyethylene glycol having bromoacetyl groups at both ends (the molecular weight of polyethylene glycol is 2000.
Abbreviated as A) 15 g of phosphate buffer 65 having a pH of 8.5
It was dissolved in 0 ml. This solution is PET-PAA / NH 2
It was poured into a glass bottle containing so that PET-PAA / NH 2 was soaked. After reacting by shaking at 25 ° C. for about 14 hours, the product was taken out and thoroughly washed with ion-exchanged water. PET-PAA / NH with PEA immobilized in this way
2 (PET-PAA-PEBA) was obtained.
【００５０】（４）リガンドの固定化 上記の操作で合成した担体に、特発性血小板減少性紫斑
病（以下ＩＴＰと略記する）の病因物質である、抗血小
板抗体との親和性を有するオリゴペプチドをリガンドと
して固定化した。このオリゴペプチドはＧｌｕ−Ｔｈｒ
−Ａｒｇ−Ａｓｎ−Ｖａｌ−Ｇｌｙ−Ｓｅｒの配列から
成るヘプタペプチド（以下Ｐｅｐと略記する）で、抗血
小板抗体に対する免疫原性がエンザイムイムノアッセイ
（以下ＥＬＩＳＡ法と略記する）により確認された。(4) Immobilization of ligand An oligopeptide having an affinity for an antiplatelet antibody, which is a causative agent of idiopathic thrombocytopenic purpura (hereinafter abbreviated as ITP), is added to the carrier synthesized by the above operation. Was immobilized as a ligand. This oligopeptide is Glu-Thr
With a heptapeptide consisting of the sequence -Arg-Asn-Val-Gly-Ser (hereinafter abbreviated as Pep), the immunogenicity against anti-platelet antibody was confirmed by an enzyme immunoassay (hereinafter abbreviated as ELISA method).
【００５１】上記のＰｅｐ４２０ｍｇをガラス瓶に取
り、ｐＨ８．５炭酸緩衝液１５０ｍｌを加えて溶解させ
た。この溶液に上記の操作で得たＰＥＴ−ＰＡＡ−ＰＥ
ＢＡ１０枚（約８．４ｇ）を浸漬し、２５℃で約１８時
間振盪して反応させた。生成物を取り出し充分に洗浄し
て、目的物であるＰｅｐ固定化ＰＥＴ製不織布ＰＥＴ−
ＰＡＡ−ＰＥＢＡ−Ｐｅｐを得た。反応残液中の窒素含
量をマイクロケルダール法で定量した値から算出したと
ころ、Ｐｅｐの固定化率は６７％であった。420 mg of the above Pep was taken in a glass bottle, and 150 ml of pH 8.5 carbonate buffer was added and dissolved. PET-PAA-PE obtained by the above operation was added to this solution.
10 sheets of BA (about 8.4 g) were immersed and shaken at 25 ° C. for about 18 hours to react. The product is taken out and thoroughly washed, and the target product, Pep-immobilized PET non-woven fabric PET-
PAA-PEBA-Pep was obtained. When the nitrogen content in the reaction residual liquid was calculated from the value quantified by the micro Kjeldahl method, the Pep immobilization rate was 67%.
【００５２】（５）リガンド固定化吸着材の吸着性能評
価 上記ＰＥＴ−ＰＡＡ−ＰＥＢＡ−Ｐｅｐを使用して評価
用モジュールを作製し、その抗血小板抗体吸着性能を、
ＩＴＰ患者の血清を用いて評価した。図１に示したのが
評価用モジュールの形態であるが、体液導入口５を有す
る漏斗型成型体３と、体液導出口６を有する漏斗型成型
体３’との間、４の部分に不織布型血液浄化材を挟み込
み、互いにネジで嵌合できるキャップ１と円筒２で締め
付ける構造になっている。このモジュールを使用し、Ｉ
ＴＰ患者の血清５０ｍｌを３７℃で３０分間潅流した。
潅流前後の血清中の抗血小板抗体量をＡｃｔａ．Ｈａｅ
ｍａｔ．，６６，２５１，１９８１に記されたＥＬＩＳ
Ａ法で定量し、吸着率を算出した。結果は表１に示した
通りであった。また、アルブミンの非特異吸着について
も潅流前後のアルブミン量をアルブミンＢ−テストワコ
ー（和光純薬工業製）で定量し、その値から吸着率を算
出した。結果は表２に示した通りであった。(5) Evaluation of Adsorption Performance of Ligand-Immobilized Adsorbent An evaluation module was prepared using the PET-PAA-PEBA-Pep, and its anti-platelet antibody adsorption performance was evaluated.
It was evaluated using the serum of ITP patients. Although the form of the evaluation module is shown in FIG. 1, between the funnel-shaped molded body 3 having the body fluid introduction port 5 and the funnel-shaped molded body 3 ′ having the body fluid discharge port 6, the nonwoven fabric is provided at the portion 4 It has a structure in which a type blood purifying material is sandwiched and tightened by a cap 1 and a cylinder 2 which can be fitted to each other with screws. Using this module, I
50 ml of TP patient serum was perfused for 30 minutes at 37 ° C.
The amount of antiplatelet antibody in the serum before and after perfusion was measured by Acta. Hae
mat. , 66, 251, 1981 ELIS
It quantified by the A method and calculated the adsorption rate. The results are as shown in Table 1. Regarding non-specific adsorption of albumin, the amount of albumin before and after perfusion was quantified with Albumin B-Test Wako (manufactured by Wako Pure Chemical Industries), and the adsorption rate was calculated from the value. The results are as shown in Table 2.
【００５３】さらに、潅流によるサンプルへの血球成分
の粘着について、東亜医用電子製血球自動計数装置Ｓｙ
ｓｍｅｘＦ−８００を用いて測定した。血球粘着につい
ては、潅流前の血液中の血球含量（白血球、血小板）に
対する、潅流後の血液中の血球含量の占める割合（血球
残存率）として算出し、表３に示した。Further, regarding adhesion of blood cell components to the sample by perfusion, automatic blood cell counter Sy manufactured by Toa Medical Electronics Co., Ltd.
It measured using smexF-800. The blood cell adhesion was calculated as the ratio (blood cell residual rate) of the blood cell content in blood after perfusion to the blood cell content (white blood cells, platelets) in blood before perfusion, and is shown in Table 3.
【００５４】[0054]
【表１】 [Table 1]
【００５５】[0055]
【表２】 [Table 2]
【００５６】[0056]
【表３】 [Table 3]
【００５７】〈実施例２〉 （１）担体の合成 繊維径８．３μｍ、目付け１２０ｇ／ｍ2 のレーヨンの
不織布を１５ｃｍ×１２ｃｍの大きさに切断した（重量
約２．１６ｇ）。このレーヨン製不織布は改質に先立
ち、アセトンで充分洗浄しておいた。過沃素酸ナトリウ
ム４０ｇをガラス瓶に取り、１規定−硫酸３．５ｌを加
えて溶解した。このガラス瓶にレーヨン製不織布１２０
枚を浸漬し、室温で約１８時間振盪して反応させた。生
成物をイオン交換水で充分洗浄し、減圧乾燥して、アル
デヒド導入レーヨン不織布（ＣＡ）を得た。アルデヒド
含量をオキシム法で定量したところ０．４１ｍｅｑ／ｇ
であった。Example 2 (1) Synthesis of Carrier A rayon nonwoven fabric having a fiber diameter of 8.3 μm and a basis weight of 120 g / m 2 was cut into a size of 15 cm × 12 cm (weight: about 2.16 g). This rayon nonwoven fabric was thoroughly washed with acetone before modification. 40 g of sodium periodate was placed in a glass bottle and 3.5 L of 1N-sulfuric acid was added to dissolve it. Rayon non-woven fabric 120 in this glass bottle
The plate was immersed and shaken at room temperature for about 18 hours for reaction. The product was thoroughly washed with ion-exchanged water and dried under reduced pressure to obtain an aldehyde-introduced rayon nonwoven fabric (CA). The aldehyde content was determined by the oxime method to be 0.41 meq / g
Met.
【００５８】上記で得たＣＡ６０枚をガラス瓶に取り、
ｐＨ９．５の炭酸緩衝液２．０ｌを加えた。これにジエ
チレントリアミン２０ｇを加え、室温で約１８時間振盪
して反応させた。生成物を取り出してイオン交換水で充
分に洗浄した後、ｐＨ９．０の炭酸緩衝液２．０ｌの入
ったビーカーに入れ、これに水素化ホウ素ナトリウム１
００ｇを加えてシッフ塩基の水素添加を行った。こうし
てアミノ基導入レーヨン不織布Ｃｅｎを得た。酸塩基滴
定でＣｅｎのアミノ基含量を定量したところ、０．３８
ｍｅｑ／ｇであった。60 CA sheets obtained above were placed in a glass bottle,
2.0 l of carbonate buffer pH 9.5 was added. 20 g of diethylenetriamine was added to this, and it was made to react by shaking at room temperature for about 18 hours. After taking out the product and thoroughly washing it with ion-exchanged water, the product was placed in a beaker containing 2.0 l of carbonate buffer having pH 9.0, and sodium borohydride 1
The Schiff base was hydrogenated by adding 00 g. Thus, an amino group-introduced rayon nonwoven fabric Cen was obtained. When the content of amino groups in Cen was quantified by acid-base titration, it was 0.38.
It was meq / g.
【００５９】ＥＤＣ１．６ｇをガラス瓶に取り、ｐＨ
４．５のリン酸緩衝液３００ｍｌを加えて溶解させた。
このガラス瓶にＰＡＡ２０ｇを加えて３０％水酸化ナト
リウム水溶液でｐＨを調節し、これに、上記で得たＣｅ
ｎ１０枚を浸漬して室温で約１８時間振盪して反応させ
た。生成物を取り出し、充分に洗浄して、ＰＡＡ固定化
レーヨン不織布Ｃ−ＰＡＡを得た。酸塩基滴定でＣ−Ｐ
ＡＡの酸含量を定量したところ、０．４０ｍｅｑ／ｇで
あった。Take 1.6 g of EDC into a glass bottle and adjust the pH.
300 ml of 4.5 phosphate buffer was added and dissolved.
To this glass bottle, 20 g of PAA was added, and the pH was adjusted with a 30% aqueous sodium hydroxide solution.
The n10 sheets were immersed and shaken at room temperature for about 18 hours for reaction. The product was taken out and thoroughly washed to obtain PAA-immobilized rayon nonwoven fabric C-PAA. CP by acid-base titration
When the acid content of AA was quantified, it was 0.40 meq / g.
【００６０】（２）末端ブロモアセチル基含有スペーサ
ーの導入 ガラス瓶に上記の操作で得たＣ−ＰＡＡ２０枚（約４
４．０ｇ）を取った。ＰＥＢＡ１４ｇを取り、ｐＨ８．
５のリン酸緩衝液９００ｍｌに溶解した。この溶液をＣ
−ＰＡＡの入ったガラス瓶に注ぎ、Ｃ−ＰＡＡが浸るよ
うにした。２５℃で約１４時間振盪して反応させた後、
不織布を取り出し、イオン交換水で充分に洗浄した。こ
うして、残存アミノ基との反応によってＰＥＢＡを固定
化したＣ−ＰＡＡ（Ｃ−ＰＡＡ−ＰＥＢＡ）を得た。(2) Introduction of terminal bromoacetyl group-containing spacer In a glass bottle, 20 sheets of C-PAA obtained by the above operation (about 4
4.0 g). Take 14 g of PEBA and adjust the pH to 8.
It was dissolved in 900 ml of the phosphate buffer solution of No. 5. This solution is C
-Pour into a glass bottle containing PAA so that C-PAA is submerged. After reacting by shaking at 25 ° C for about 14 hours,
The nonwoven fabric was taken out and thoroughly washed with ion-exchanged water. Thus, C-PAA (C-PAA-PEBA) having PEBA immobilized by the reaction with the residual amino group was obtained.
【００６１】（３）リガンドの固定化 上記で得たＣ−ＰＡＡ−ＰＥＢＡに実施例１でＰＥＴ−
ＰＡＡ−ＰＥＢＡにＰｅｐを導入したのと同様の方法で
Ｐｅｐを固定化してＣ−ＰＡＡ−ＰＥＢＡ−Ｐｅｐを得
た。反応残液中の窒素含量をマイクロケルダール法で定
量した値から算出したところ、Ｐｅｐの固定化率は６８
％であった。(3) Immobilization of ligand PET-in Example 1 was added to the C-PAA-PEBA obtained above.
Pep was immobilized by the same method as that for introducing Pep into PAA-PEBA to obtain C-PAA-PEBA-Pep. When the nitrogen content in the reaction residual liquid was calculated from the value quantified by the micro Kjeldahl method, the immobilization rate of Pep was 68.
%Met.
【００６２】（４）リガンド固定化吸着材の吸着性能評
価 実施例１と同様の方法でＣ−ＰＡＡ−ＰＥＢＡ−Ｐｅｐ
の抗血小板抗体吸着性能、アルブミン非特異吸着、潅流
後の血球残存率について評価した。結果はそれぞれ表
１、表２、表３に示した。(4) Evaluation of adsorption performance of ligand-immobilized adsorbent C-PAA-PEBA-Pep was prepared in the same manner as in Example 1.
Was evaluated for anti-platelet antibody adsorption performance, non-specific albumin adsorption, and blood cell survival rate after perfusion. The results are shown in Table 1, Table 2 and Table 3, respectively.
【００６３】〈比較例１〉 （１）担体の合成 実施例２で得た、アルデヒド含量０．４１ｍｅｑ／ｇの
ＣＡをそのまま担体として用いた。Comparative Example 1 (1) Synthesis of Carrier The CA having an aldehyde content of 0.41 meq / g obtained in Example 2 was used as it was as a carrier.
【００６４】（２）リガンドの導入 ＣＡ１０枚をガラス瓶に取り、ｐＨ９．５の炭酸緩衝液
４００ｍｌを加えた。実施例１で使用したのと同じヘプ
タペプチドＰｅｐをあらかじめ少量のｐＨ９．５炭酸緩
衝液に溶解しておき、この溶液をＣＡの入った容器に加
え、室温で約１８時間振盪して反応させた。生成物を取
り出してイオン交換水で充分に洗浄した後、ｐＨ９．０
の炭酸緩衝液４００ｍｌの入ったビーカーに入れ、これ
に水素化ホウ素ナトリウム２０ｇを加えてシッフ塩基の
水素添加を行った。生成物を取り出し、充分に洗浄し
て、目的物であるＰｅｐ固定化レーヨン不織布Ｃ−Ｐｅ
ｐを得た。反応残液中の窒素含量をマイクロケルダール
法で定量した値から算出したところ、Ｐｅｐの固定化率
は７８％であった。(2) Introduction of ligand Ten CA sheets were taken in a glass bottle and 400 ml of a carbonate buffer solution having a pH of 9.5 was added. The same heptapeptide Pep used in Example 1 was previously dissolved in a small amount of a pH 9.5 carbonate buffer solution, this solution was added to a container containing CA, and the mixture was reacted by shaking at room temperature for about 18 hours. . After taking out the product and thoroughly washing it with ion-exchanged water, pH 9.0
The solution was placed in a beaker containing 400 ml of the carbonate buffer solution, and 20 g of sodium borohydride was added to the beaker to hydrogenate the Schiff base. The product is taken out and thoroughly washed to obtain the desired product, Pep-immobilized rayon nonwoven fabric C-Pe.
got p. When the nitrogen content in the reaction residual liquid was calculated from the value quantified by the micro Kjeldahl method, the Pep immobilization rate was 78%.
【００６５】（３）リガンド固定化吸着材の吸着性能評
価 実施例１と同様の方法でＣ−Ｐｅｐの抗血小板抗体吸着
性能、アルブミン非特異吸着、潅流後の血球残存率につ
いて評価した。結果はそれぞれ表１、表２、表３に示し
た。(3) Evaluation of adsorption performance of ligand-immobilized adsorbent In the same manner as in Example 1, the anti-platelet antibody adsorption performance of C-Pep, albumin non-specific adsorption, and blood cell survival rate after perfusion were evaluated. The results are shown in Table 1, Table 2 and Table 3, respectively.
【００６６】〈比較例２〉 （１）担体の合成 実施例２で得た、アルデヒド含量０．４１ｍｅｑ／ｇの
ＣＡ６０枚をガラス瓶に取り、ｐＨ９．５の炭酸緩衝液
２．５ｌを加えた。これにＬ−グルタミン酸２０ｇを加
え、室温で約１８時間振盪して反応させた。生成物を取
り出してイオン交換水で充分に洗浄した後、ｐＨ９．０
の炭酸緩衝液１．２ｌの入ったビーカーに入れ、これに
水素化ホウ素ナトリウム１００ｇを加えてシッフ塩基の
水素添加を行った。こうしてカルボキシル基導入レーヨ
ン不織布ＣＧを得た。酸塩基滴定でＣＧのカルボキシル
基含量を定量したところ、０．３６ｍｅｑ／ｇであっ
た。Comparative Example 2 (1) Synthesis of Carrier 60 sheets of CA having an aldehyde content of 0.41 meq / g obtained in Example 2 were placed in a glass bottle and 2.5 l of a carbonate buffer solution having a pH of 9.5 was added. 20 g of L-glutamic acid was added to this, and it was made to react by shaking at room temperature for about 18 hours. After taking out the product and thoroughly washing it with ion-exchanged water, pH 9.0
Into a beaker containing 1.2 liters of the carbonate buffer solution (1) was added 100 g of sodium borohydride, and the Schiff base was hydrogenated. Thus, a carboxyl group-introduced rayon nonwoven fabric CG was obtained. The carboxyl group content of CG was determined by acid-base titration to be 0.36 meq / g.
【００６７】（２）リガンドの導入 ＥＤＣ２２０ｍｇをガラス瓶に取り、ｐＨ４．５のリン
酸緩衝液４００ｍｌを加えて溶解した。このガラス瓶に
Ｐｅｐを加えて溶解させた後ＣＧ１０枚（約２２．１
ｇ）を浸漬し、室温で約１８時間振盪して反応させた。
生成物を取り出し、充分に洗浄して、目的物であるＰｅ
ｐ固定化レーヨン不織布ＣＧ−Ｐｅｐを得た。反応残液
中の窒素含量をマイクロケルダール法で定量した値から
算出したところ、Ｐｅｐの固定化率は７２％であった。(2) Introduction of ligand 220 mg of EDC was placed in a glass bottle and 400 ml of a phosphate buffer having a pH of 4.5 was added and dissolved. After adding Pep to the glass bottle to dissolve it, 10 CGs (about 22.1
g) was immersed and shaken at room temperature for about 18 hours to react.
Take out the product, wash it thoroughly, and remove the target Pe.
A p-immobilized rayon nonwoven fabric CG-Pep was obtained. When the nitrogen content in the reaction residual liquid was calculated from the value quantified by the micro Kjeldahl method, the Pep immobilization rate was 72%.
【００６８】（３）リガンド固定化吸着材の吸着性能評
価 実施例１と同様の方法でＣＧ−Ｐｅｐの抗血小板抗体吸
着性能、アルブミン非特異吸着、潅流後の血球残存率に
ついて評価した。結果はそれぞれ表１、表２、表３に示
した。(3) Evaluation of adsorption performance of ligand-immobilized adsorbent In the same manner as in Example 1, CG-Pep was evaluated for anti-platelet antibody adsorption performance, albumin non-specific adsorption, and blood cell survival rate after perfusion. The results are shown in Table 1, Table 2 and Table 3, respectively.
【００６９】〈比較例３〉 （１）担体の合成 実施例１で得たカルボキシル基含量０．３２ｍｅｑ／ｇ
のＰＥＴ−ＰＡＡをそのまま担体として用いた。Comparative Example 3 (1) Synthesis of Carrier Carboxyl group content obtained in Example 1 0.32 meq / g
PET-PAA of was used as a carrier as it was.
【００７０】（２）リガンドの固定化 ＥＤＣ８０ｍｇをガラス瓶に取り、ｐＨ４．５のリン酸
緩衝液１５０ｍｌを加えて溶解した。このガラス瓶にＰ
ｅｐを加えて溶解させた後ＰＥＴ−ＰＡＡ１０枚（約
８．１ｇ）を浸漬し、室温で約１８時間振盪して反応さ
せた。生成物を取り出し、充分に洗浄して、目的物であ
るＰｅｐ固定化ＰＥＴ製不織布ＰＥＴ−ＰＡＡ−Ｐｅｐ
を得た。反応残液中の窒素含量をマイクロケルダール法
で定量した値から算出したところ、Ｐｅｐの固定化率は
７７％であった。(2) Immobilization of ligand 80 mg of EDC was placed in a glass bottle, and 150 ml of phosphate buffer having a pH of 4.5 was added and dissolved. P in this glass bottle
After ep was added and dissolved, 10 pieces of PET-PAA (about 8.1 g) were immersed, and reacted by shaking at room temperature for about 18 hours. The product is taken out, washed thoroughly, and the target product, Pep-immobilized PET non-woven fabric PET-PAA-Pep.
Got When the nitrogen content in the reaction residual liquid was calculated from the value quantified by the micro Kjeldahl method, the Pep immobilization rate was 77%.
【００７１】（３）リガンド固定化吸着材の吸着性能評
価 実施例１と同様の方法でＰＥＴ−ＰＡＡ−Ｐｅｐの抗血
小板抗体吸着性能、アルブミン非特異吸着、潅流後の血
球残存率について評価した。結果はそれぞれ表１、表
２、表３に示した。(3) Evaluation of adsorption performance of ligand-immobilized adsorbent In the same manner as in Example 1, PET-PAA-Pep was evaluated for anti-platelet antibody adsorption performance, albumin non-specific adsorption, and blood cell residual rate after perfusion. The results are shown in Table 1, Table 2 and Table 3, respectively.
【００７２】上記の例から明らかなように、本発明の不
織布型血液浄化吸着材は血中の特定成分（抗血小板抗
体）を選択的、かつ簡便に除去できた。担体上にポリカ
ルボン酸が存在しない場合（比較例１）では抗血小板抗
体の吸着能が大きく劣っている。また、素材が同一であ
る実施例２と比較した場合、アルブミンの非特異吸着や
処理後の血球残存率についても本発明の吸着材に劣って
いる。また、担体上に低分子のカルボキシル基含有化合
物（Ｌ−グルタミン酸）を導入した場合（比較例２）
は、アルブミン非特異吸着や処理後の血球残存率につい
ては本発明の不織布型血液浄化吸着材と比較しても大き
く劣ってはいないが、抗血小板抗体の吸着能はいまだ満
足できるレベルではない。担体上にポリカルボン酸が存
在し、親水性スペーサーを介さずにリガンドを固定した
場合（比較例３）、比較例１、２と比べると血液適合
性、吸着性能ともかなり改善されてはいるが、いまだ充
分とは言い難い。As is clear from the above example, the non-woven blood purification adsorbent of the present invention was able to selectively and easily remove a specific component (antiplatelet antibody) in blood. In the case where no polycarboxylic acid is present on the carrier (Comparative Example 1), the anti-platelet antibody adsorption ability is greatly inferior. In addition, when compared with Example 2 in which the materials are the same, the nonspecific adsorption of albumin and the blood cell residual rate after treatment are also inferior to the adsorbent of the present invention. Further, when a low molecular weight carboxyl group-containing compound (L-glutamic acid) is introduced onto the carrier (Comparative Example 2)
The non-specific adsorption of albumin and the blood cell residual rate after treatment are not significantly inferior to those of the non-woven blood purification adsorbent of the present invention, but the anti-platelet antibody adsorption capacity is still not at a satisfactory level. When a polycarboxylic acid is present on a carrier and a ligand is immobilized without a hydrophilic spacer (Comparative Example 3), both blood compatibility and adsorption performance are considerably improved as compared with Comparative Examples 1 and 2. It's still hard to say.
【００７３】比較例１〜３と比べ、分子量４００〜４０
０００のポリカルボン酸を担体上に導入し、さらに両末
端にブロモアセチル基を有する親水性スペーサーを介し
てリガンドを固定した実施例のデータは、優れた血液適
合性と吸着性能が両立されている。As compared with Comparative Examples 1 to 3, the molecular weight is 400 to 40.
The data of the example in which 000 polycarboxylic acids were introduced on the carrier and the ligand was immobilized via hydrophilic spacers having bromoacetyl groups at both ends, excellent blood compatibility and adsorption performance are compatible. .
【００７４】[0074]
【発明の効果】本発明の不織布型血液浄化吸着材は、分
子量４００〜４００００のポリカルボン酸が持つ負電荷
と、非イオン性親水性スペーサーの働きにより、良好な
吸着性能と血液適合性を両立することができる。また、
担体の形状が不織布型であるため全血を潅流した際の圧
損を低く抑えることが可能となり、あらかじめ血液から
血漿成分を分離して潅流するという煩雑な方法をとらず
直接血液を潅流することによって充分な効果を期待する
ことができる。このため、簡便な操作で、血液中の病因
物質の除去による種々の疾病の治療、或いはＢ細胞、Ｔ
細胞等特定成分の選択的な分離に広く利用され得る。INDUSTRIAL APPLICABILITY The non-woven blood purification adsorbent of the present invention has both good adsorption performance and blood compatibility due to the negative charge of the polycarboxylic acid having a molecular weight of 400 to 40,000 and the function of the nonionic hydrophilic spacer. can do. Also,
Since the shape of the carrier is a non-woven type, it is possible to suppress the pressure loss when perfusing whole blood to a low level, and by directly perfusing blood without the complicated method of separating plasma components from blood and perfusing in advance. A sufficient effect can be expected. For this reason, various diseases can be treated by removing pathogenic substances in blood, or B cells, T
It can be widely used for selective separation of specific components such as cells.
【図１】実施例で使用される評価用モジュールの形態つ
いて図示したものである。FIG. 1 illustrates a form of an evaluation module used in an example.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 田中 昌和 滋賀県大津市堅田二丁目１番１号 東洋紡 績株式会社総合研究所内 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Masakazu Tanaka 2-1-1 Katata, Otsu City, Shiga Prefecture Toyobo Co., Ltd.
Claims (3)
のポリカルボン酸を導入した繊維径０．１〜１００μｍ
の繊維から成る水不溶性不織布に、両末端にブロモアセ
チル基を有する親水性スペーサーを介して、被吸着物質
と親和性を有する有機化合物を固定して成る血液浄化吸
着材。1. A molecular weight of 400 to 40,000 as a carrier.
Diameter of the fiber introduced with the polycarboxylic acid of 0.1 to 100 μm
A blood purification adsorbent comprising a water-insoluble non-woven fabric composed of the above fibers and an organic compound having an affinity for the substance to be adsorbed, which is immobilized via hydrophilic spacers having bromoacetyl groups at both ends.
カルボキシル基含量が１０〜１０4 μｅｑ／ｇであるこ
とを特徴とする特許請求の範囲第１項記載の血液浄化吸
着材。2. The blood purification adsorbent according to claim 1, wherein the content of the carboxyl group derived from the polycarboxylic acid on the surface of the carrier is 10 to 10 4 μeq / g.
性スペーサーが、下記化１の構造を有する化合物である
ことを特徴とする特許請求の範囲第１項記載の血液浄化
吸着材。化１において、 ｎ＝０〜１０の整数 ｍ＝２〜４００の整数 ｌ＝１ま
たは２ Ｒ1 、Ｒ2 は水素原子またはメチル基で、それぞれ同じ
もしくは異なってもよい。ＸはＮＨ、Ｏ、またはＣＨ2
を各々示す。 【化１】 3. The blood purification adsorbent according to claim 1, wherein the hydrophilic spacer having bromoacetyl groups at both ends is a compound having the structure of the following chemical formula 1. In Chemical formula 1, n = 0 to 10 integer m = 2 to 400 integer l = 1 or 2 R 1 and R 2 are hydrogen atoms or methyl groups, and may be the same or different. X is NH, O, or CH 2
Are shown respectively. [Chemical 1]
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP50A JPH06233815A (en) | 1993-02-10 | 1993-02-10 | Hemocatharsis adsorbent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP50A JPH06233815A (en) | 1993-02-10 | 1993-02-10 | Hemocatharsis adsorbent |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH06233815A true JPH06233815A (en) | 1994-08-23 |
Family
ID=12088414
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP50A Pending JPH06233815A (en) | 1993-02-10 | 1993-02-10 | Hemocatharsis adsorbent |
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| Country | Link |
|---|---|
| JP (1) | JPH06233815A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6582386B2 (en) | 2001-03-06 | 2003-06-24 | Baxter International Inc. | Multi-purpose, automated blood and fluid processing systems and methods |
| US6706008B2 (en) | 2001-03-06 | 2004-03-16 | Baxter International Inc. | Automated system and method for withdrawing compounds from blood |
| US6884228B2 (en) | 2001-03-06 | 2005-04-26 | Baxter International Inc. | Automated system adaptable for use with different fluid circuits |
-
1993
- 1993-02-10 JP JP50A patent/JPH06233815A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6582386B2 (en) | 2001-03-06 | 2003-06-24 | Baxter International Inc. | Multi-purpose, automated blood and fluid processing systems and methods |
| US6706008B2 (en) | 2001-03-06 | 2004-03-16 | Baxter International Inc. | Automated system and method for withdrawing compounds from blood |
| US6884228B2 (en) | 2001-03-06 | 2005-04-26 | Baxter International Inc. | Automated system adaptable for use with different fluid circuits |
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