JPS63139901A - Heparin derivative and its preparation - Google Patents
Heparin derivative and its preparationInfo
- Publication number
- JPS63139901A JPS63139901A JP28666186A JP28666186A JPS63139901A JP S63139901 A JPS63139901 A JP S63139901A JP 28666186 A JP28666186 A JP 28666186A JP 28666186 A JP28666186 A JP 28666186A JP S63139901 A JPS63139901 A JP S63139901A
- Authority
- JP
- Japan
- Prior art keywords
- heparin
- epoxy
- reaction
- epoxy groups
- epoxy compound
- 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
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical class OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 title claims abstract description 81
- 239000002628 heparin derivative Substances 0.000 title claims description 26
- 229920000669 heparin Polymers 0.000 claims abstract description 56
- 229960002897 heparin Drugs 0.000 claims abstract description 55
- 125000003700 epoxy group Chemical group 0.000 claims abstract description 35
- 150000001875 compounds Chemical class 0.000 claims abstract description 28
- 239000004593 Epoxy Substances 0.000 claims abstract description 23
- 239000007788 liquid Substances 0.000 claims abstract description 3
- 238000006243 chemical reaction Methods 0.000 claims description 25
- 239000007795 chemical reaction product Substances 0.000 claims description 11
- 238000004519 manufacturing process Methods 0.000 claims description 11
- 239000007864 aqueous solution Substances 0.000 claims description 10
- 125000003277 amino group Chemical group 0.000 claims description 9
- SHKUUQIDMUMQQK-UHFFFAOYSA-N 2-[4-(oxiran-2-ylmethoxy)butoxymethyl]oxirane Chemical compound C1OC1COCCCCOCC1CO1 SHKUUQIDMUMQQK-UHFFFAOYSA-N 0.000 claims description 2
- 229920002678 cellulose Polymers 0.000 abstract description 16
- 239000001913 cellulose Substances 0.000 abstract description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 12
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 abstract description 9
- 230000002785 anti-thrombosis Effects 0.000 abstract description 7
- WERYXYBDKMZEQL-UHFFFAOYSA-N butane-1,4-diol Chemical compound OCCCCO WERYXYBDKMZEQL-UHFFFAOYSA-N 0.000 abstract description 7
- 230000000694 effects Effects 0.000 abstract description 7
- 239000003146 anticoagulant agent Substances 0.000 abstract description 5
- 239000000463 material Substances 0.000 abstract description 5
- 239000003960 organic solvent Substances 0.000 abstract description 2
- 239000004615 ingredient Substances 0.000 abstract 2
- 239000012295 chemical reaction liquid Substances 0.000 abstract 1
- 239000000126 substance Substances 0.000 abstract 1
- 238000000034 method Methods 0.000 description 13
- 239000000243 solution Substances 0.000 description 10
- 239000012567 medical material Substances 0.000 description 6
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 230000035484 reaction time Effects 0.000 description 4
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 208000007536 Thrombosis Diseases 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 230000014508 negative regulation of coagulation Effects 0.000 description 3
- 229920000642 polymer Polymers 0.000 description 3
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000002429 anti-coagulating effect Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- SFKTYEXKZXBQRQ-UHFFFAOYSA-J thorium(4+);tetrahydroxide Chemical compound [OH-].[OH-].[OH-].[OH-].[Th+4] SFKTYEXKZXBQRQ-UHFFFAOYSA-J 0.000 description 2
- AOBIOSPNXBMOAT-UHFFFAOYSA-N 2-[2-(oxiran-2-ylmethoxy)ethoxymethyl]oxirane Chemical compound C1OC1COCCOCC1CO1 AOBIOSPNXBMOAT-UHFFFAOYSA-N 0.000 description 1
- FZZMTSNZRBFGGU-UHFFFAOYSA-N 2-chloro-7-fluoroquinazolin-4-amine Chemical group FC1=CC=C2C(N)=NC(Cl)=NC2=C1 FZZMTSNZRBFGGU-UHFFFAOYSA-N 0.000 description 1
- 229920002683 Glycosaminoglycan Polymers 0.000 description 1
- 241000609816 Pantholops hodgsonii Species 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 239000004721 Polyphenylene oxide Substances 0.000 description 1
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 1
- 238000011481 absorbance measurement Methods 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 238000000149 argon plasma sintering Methods 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000010908 decantation Methods 0.000 description 1
- GYZLOYUZLJXAJU-UHFFFAOYSA-N diglycidyl ether Chemical compound C1OC1COCC1CO1 GYZLOYUZLJXAJU-UHFFFAOYSA-N 0.000 description 1
- ZFGMDIBRIDKWMY-PASTXAENSA-N heparin Chemical compound CC(O)=N[C@@H]1[C@@H](O)[C@H](O)[C@@H](COS(O)(=O)=O)O[C@@H]1O[C@@H]1[C@@H](C(O)=O)O[C@@H](O[C@H]2[C@@H]([C@@H](OS(O)(=O)=O)[C@@H](O[C@@H]3[C@@H](OC(O)[C@H](OS(O)(=O)=O)[C@H]3O)C(O)=O)O[C@@H]2O)CS(O)(=O)=O)[C@H](O)[C@H]1O ZFGMDIBRIDKWMY-PASTXAENSA-N 0.000 description 1
- 229960001008 heparin sodium Drugs 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-M hydroxide Chemical compound [OH-] XLYOFNOQVPJJNP-UHFFFAOYSA-M 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- BSCHIACBONPEOB-UHFFFAOYSA-N oxolane;hydrate Chemical compound O.C1CCOC1 BSCHIACBONPEOB-UHFFFAOYSA-N 0.000 description 1
- 229920000570 polyether Polymers 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- KKCBUQHMOMHUOY-UHFFFAOYSA-N sodium oxide Chemical compound [O-2].[Na+].[Na+] KKCBUQHMOMHUOY-UHFFFAOYSA-N 0.000 description 1
- 229910001948 sodium oxide Inorganic materials 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- -1 sulfoamino group Chemical group 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 229950003937 tolonium Drugs 0.000 description 1
Landscapes
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
Description
【発明の詳細な説明】 （産業上の利用分野） 本発明はヘパリン誘導体及びその製造法に関する。[Detailed description of the invention] (Industrial application field) The present invention relates to heparin derivatives and methods for producing the same.
更に詳しくは、本ブし明は、ヘパリンとエポキシ化合物
との反応生成物であるヘパリン誘導体とモの製造法に関
するものである。More specifically, this patent relates to a method for producing heparin derivatives, which are reaction products of heparin and epoxy compounds.
本発明のヘパリン誘導体は、ヘパリンの活性が失なわれ
ず、かつセルロースのような高分子と容易に結合し１７
るものであるため、抗血栓性材料の製造に用いる中間体
としてきわめて有用である。The heparin derivative of the present invention does not lose heparin activity and easily binds to polymers such as cellulose.
Therefore, it is extremely useful as an intermediate used in the production of antithrombotic materials.
（従来の技術）
ヘパリンを利用して医用材料に抗血栓性を与える方法が
従来より知られている。例えば、特開昭５８−１００５
３号公報に開示されているように共有結合を形成ざＶて
医用材料とヘパリンを結合ざぜる方法がある。(Prior Art) A method of imparting antithrombotic properties to medical materials using heparin is conventionally known. For example, JP-A-58-1005
As disclosed in Japanese Patent Application No. 3, there is a method of combining a medical material and heparin by forming a covalent bond.
しかし、この方法はヘパリン活性サイトも反応に関与す
る可能性があるため、ヘパリンの抗血液凝固活性が失な
われて血栓を発生させる可能性があり望ましくない。ま
た、特開昭４９−８５８３号及び特開昭４９−４４５９
０号公報に開示されているように、イオン結合によって
ヘパリンを結合させる方法も知られているが、医用材料
とヘパリンとの結合が弱いため、ヘパリンが容易に剥れ
やすく、長期にわたる使用には適しなかった。However, this method is undesirable because heparin active sites may also be involved in the reaction, which may result in the loss of heparin's anticoagulant activity and the generation of thrombus. Also, JP-A-49-8583 and JP-A-49-4459
As disclosed in Publication No. 0, a method of bonding heparin through ionic bonding is also known, but since the bond between medical materials and heparin is weak, heparin easily peels off and is not suitable for long-term use. It wasn't suitable.
（発明が解決しようとする問題点）
本発明は、上記従来技術の問題点を解決せんとするもの
であり、ヘパリンの抗血液凝固活性が低下されずしかも
セルロール等の高分子からなる医用材料に結合しｊｑる
得るヘパリンＸ４体及びその製造法を提供することを目
的とする。(Problems to be Solved by the Invention) The present invention aims to solve the above-mentioned problems of the prior art, and it is an object of the present invention to solve the above-mentioned problems of the prior art. The object of the present invention is to provide a heparin
（問題点を解決するための手段）
上記目的を達成するため、本ブを明は下記の溝底を有す
る。(Means for solving the problem) In order to achieve the above object, this plate has the following groove bottom.
（１）ヘパリンと２個以上のエポキシ基を有するエポキ
シ化合物との反応生成物でおって、ヘパリンのアミノ基
とエポキシ化合物のエポキシ基とが結合し、かつ反応生
成物中に未反応のエポキシ基が残存していることを特徴
とするヘパリン誘導体。(1) A reaction product of heparin and an epoxy compound having two or more epoxy groups, in which the amino group of heparin and the epoxy group of the epoxy compound are bonded, and there are unreacted epoxy groups in the reaction product. A heparin derivative characterized by remaining.
（２）エポキシ化合物がジグリシジル化合物であること
を特徴とする第１項に記載のヘパリン誘導体。(2) The heparin derivative according to item 1, wherein the epoxy compound is a diglycidyl compound.
（３）ジグリシジル化合物が１，４−ブタンジオールシ
グリシジルエーテルでおることを１４１″！ｉとする第
２項に記載のヘパリン誘導体。(3) The heparin derivative according to item 2, wherein 141''!i indicates that the diglycidyl compound is 1,4-butanediolsiglycidyl ether.
（４）ヘパリンと２個以上のエポキシ基を有するエポキ
シ化合物との反応生成物で必って、へバリンのアミノ基
とエポキシ化合物のエポキシ基とが結合し、かつ反応生
成物中に未反応のエポキシ基が残存しているヘパリン誘
導体を製造するにあたり、ヘパリンと２個以上のエポキ
シ基を有するエポキシ化合物とをｐＨ７以上の液体中で
反応させることを特徴とするヘパリン誘導体の製造法。(4) In the reaction product of heparin and an epoxy compound having two or more epoxy groups, the amino group of heparin and the epoxy group of the epoxy compound are bound, and the reaction product contains unreacted A method for producing a heparin derivative, which comprises reacting heparin with an epoxy compound having two or more epoxy groups in a liquid having a pH of 7 or higher, in producing a heparin derivative in which an epoxy group remains.
（５）ｐｌ−１は８〜１０である第４項記載の製造法。(5) The manufacturing method according to item 4, wherein pl-1 is 8 to 10.
（６）ｐＨ９の水）ｄ液中至温で反応させることを特徴
とする第４項記載の製造法。(6) The production method according to item 4, characterized in that the reaction is carried out at the lowest temperature in a solution (water) having a pH of 9.
水弁明番こ使用されるヘパリンとは、血液凝固阻止作用
を有するムコ多糖類の１種をいい、筋肉、肺、ｐＩＩｌ
ｌｌａ、肝臓において多く存在し、血栓症等の治療に使
用されている。上記したように、各種の医用材料にヘパ
リンを被覆して、該医用材料を生体に用いた場合に血栓
が生成するのを防止せんとする試みがなされている。The heparin used in Mizubenmeibanko is a type of mucopolysaccharide that has anticoagulant effects, and is used in muscle, lung, and pII.
lla, is present in large amounts in the liver and is used to treat thrombosis, etc. As mentioned above, attempts have been made to coat various medical materials with heparin to prevent the formation of blood clots when the medical materials are used in living organisms.
本発明において使用されるエポキシ化合物は、エポキシ
基を２個１ス上有することを要し、好適な例としてジグ
リシジル化合物があげられ、１，４−ブタンジオールシ
グリシジルエーテル、ポリエーテルジグリシジエーテル
及びポリエチレングリコールジグリシジルエーテルが好
ましく、特に１，４−ブタンジオールシグリシジルエー
テルが好ましい。The epoxy compound used in the present invention must have two epoxy groups, and preferred examples include diglycidyl compounds, such as 1,4-butanediol siglycidyl ether, polyether diglycidyl ether, and Polyethylene glycol diglycidyl ether is preferred, and 1,4-butanediol diglycidyl ether is particularly preferred.
本発明のヘパリン誘導体は１分子当り１〜１０個の未反
応エポキシ基を有しているのが好ましい。The heparin derivative of the present invention preferably has 1 to 10 unreacted epoxy groups per molecule.
本発明によるヘパリン誘導体を製造するにａｔこり、ヘ
パリンとエポキシ化合物の溶液を合わせて反応させる。To prepare the heparin derivative according to the invention, solutions of heparin and an epoxy compound are combined and reacted.
ヘパリンとエポキシ化合物の使用割合はおよそモル比で
１：４〜１：１０か望ましい。The ratio of heparin and epoxy compound to be used is preferably approximately 1:4 to 1:10 in molar ratio.
反応液のｐｌ−１は７以上とし、好ましく（よＩ） ｌ
−！ ７〜］２であり、更に好ましくはｐＨ８〜］Ｏで
市る。The pl-1 of the reaction solution is 7 or more, preferably (yoI) l
-! 7-]2, more preferably pH8-]O.
ｐｌ−１が７未満であると、ヘパリン分子中のスルホア
ミノ基が破壊され、ヘパリンのもつ抗血栓性が示されな
くなるばかりでなく、エポキシ化合物中のエポキシ基が
開環してしまい、反応そのものか起Ｖなくなるので好ま
しくない。If pl-1 is less than 7, the sulfoamino group in the heparin molecule is destroyed, and not only does heparin no longer exhibit its antithrombotic properties, but the epoxy group in the epoxy compound opens, causing the reaction itself to deteriorate. This is not preferable because it eliminates V.
反応は反応液が凝固しない温度以上１５０℃以下の温度
で実施される。１ｂＯ’Ｃより高い温１良ではヘパリン
分子が崩壊してしまうので適当でない。The reaction is carried out at a temperature above 150° C. at which the reaction solution does not solidify. Temperatures higher than 1bO'C are not suitable because heparin molecules will disintegrate.
ここでｐＨおよび反応温度がともに高いと反応性が高く
なり過ぎてエポキシ化合物中に存在する全てのエポキシ
基がヘパリンと結合してしまい、高分子医用材料との結
合が不可能となるばかりでなく、ヘパリン中の７ミノ基
以外の基との結合が生じることによるヘパリンの力価の
低下、またヘパリン自体の分解が生じる可ｌｉｔ性があ
る。従ってｐＨが高い場合には反応温度を低くし、反応
温度が高い場合にははｐＨを下げて反応させる必要があ
る。If both the pH and the reaction temperature are high, the reactivity will become too high and all the epoxy groups present in the epoxy compound will bond with heparin, making it not only impossible to bond with the polymeric medical material. , a decrease in the potency of heparin due to the formation of a bond with a group other than the 7-mino group in heparin, and a possibility that heparin itself may be degraded. Therefore, when the pH is high, it is necessary to lower the reaction temperature, and when the reaction temperature is high, it is necessary to lower the pH for the reaction.
反応時間はｐ（−１や反応温度等の他の反応条件にＪ：
つて異なるが一般に３０分間ないし１０日間である。例
えばｐＨ９のとき、６０℃で２４時間、至温で４日間反
応させるのが適当である。The reaction time is J depending on other reaction conditions such as p(-1 and reaction temperature.
The time varies, but generally ranges from 30 minutes to 10 days. For example, when the pH is 9, it is appropriate to react at 60° C. for 24 hours and at the lowest temperature for 4 days.
反応は水、水溶液または親水性０機溶媒またはそれらの
混合物の存在下で実ｒ＃、される。好適には水酸化す（
〜リウムもしくは水酸化カリウム水溶液、リン酸もしく
はホウ酸系バッフ１溶液、テトラヒドロフラン系溶媒と
水との混合ｒｉ＜例えば゛アトラヒドロフラン：水−３
：１）、ジオキサン系溶媒と水との混合液（例えばジオ
キサン：水−９：１）笠か使用される。このように各反
応条件を調和な条件とすることによりヘパリンの抗血液
凝固活性発現に直接関与しないアミノ基のみをエポキシ
基と反応させることかでき、かつエポキシ化合物に存在
する全てのエポキシ基がヘパリンと結合するのを防止す
ることができる。その結果、本発明によるヘパリン誘導
体は、未反応のエポキシ基を有し、その未反応エポキシ
基を利用して、セルロース等の高分子中に存在する水酸
基等と結合させることによって抗血栓性材料を製造せん
とするものであり、このような抗血栓性材料の製造にお
ける中間体として有用である。The reaction is carried out in the presence of water, an aqueous solution or a hydrophilic organic solvent or a mixture thereof. Preferably hydroxide (
~ Lium or potassium hydroxide aqueous solution, 1 solution of phosphoric acid or boric acid buff, a mixture of tetrahydrofuran solvent and water
:1), a mixed solution of a dioxane-based solvent and water (for example, dioxane:water - 9:1) is used. By harmonizing each reaction condition in this way, only the amino groups that are not directly involved in the expression of heparin's anticoagulant activity can be reacted with epoxy groups, and all the epoxy groups present in the epoxy compound can be reacted with heparin. can be prevented from combining with As a result, the heparin derivative according to the present invention has unreacted epoxy groups, and utilizes the unreacted epoxy groups to combine with hydroxyl groups present in polymers such as cellulose, thereby forming antithrombotic materials. It is intended to be manufactured and is useful as an intermediate in the manufacture of such antithrombotic materials.
（実施例） 以下実施例により本発明をさらに詳しく説明する。(Example) The present invention will be explained in more detail with reference to Examples below.
実施例１
１．４ブタンジオールジグリシジルエーテル０．０８１
９（４ｘ１０−４ｍｏ Ｉ ）を１０ｄの水に溶解し、
これにヘパリンナトリウム１．０３を加え均一に溶解ざ
ぜたのら、０．１Ｎ水酸化ナトリウム溶液で反応溶液の
ｐＨを９．０に調整した。Example 1 1.4 Butanediol diglycidyl ether 0.081
9 (4x10-4 mo I) in 10 d of water,
After adding 1.03 ml of heparin sodium and dissolving it uniformly, the pH of the reaction solution was adjusted to 9.0 with 0.1N sodium hydroxide solution.
この溶液を至温で４日間撹拌反応させ、０．１Ｎ塩酸に
てｐＨを７．０に調整したのち、未反応１．４ブタンジ
オールジグリシジルエーテルを除くため１０！ｄ！によ
り３回抽出し、残ったヘパリン誘導体水溶液を軽く蒸発
させた後所定量に水で希釈した。This solution was stirred and reacted at the lowest temperature for 4 days, and after adjusting the pH to 7.0 with 0.1N hydrochloric acid, 10! d! The remaining heparin derivative aqueous solution was lightly evaporated and diluted with water to a predetermined amount.
上記のヘパリン誘導体合成時に６ける１、４ブタンジオ
ールジグリシジルエーデルの減少量を高速液体クロマ１
〜グラフイーで調べたところ、第１図にポリ“ように反
応４日日で反応率６０％に達していた。一方、原料ヘパ
リン中にはアミン基が３〜４個存在づることが知られて
おり（Ｕ、 Ｌｉ、ｎｄａｈｌ−ｅｔａｌ、Ｊ、Ｂｉｏ
ｌ、Ｃｈｅｍ、、２４８．７２３４（１９７３）） 、
このことをニンヒドリン法で６Ｋ 認したところ、従来
の結果と一致した。また、光散乱法によるヘパリンの分
子単測定では、本ヘパリンの分子間は１３０００という
結果か冑られた。High performance liquid chromatography 1
~ When examined using graphie, the reaction rate reached 60% in 4 days as shown in Figure 1, which shows that the reaction rate reached 60% in 4 days.On the other hand, it is known that there are 3 to 4 amine groups in the raw material heparin. (U, Li, ndahl-etal, J, Bio
Chem, 248.7234 (1973)),
This was confirmed using the ninhydrin method using the 6K method, and the result was consistent with conventional results. Furthermore, in a single molecule measurement of heparin using a light scattering method, the number between molecules of this heparin was 13,000, which was shocking.
これらの事実から本合成によるヘパリン誘導体１分子当
り約３個のエポキシ基か存在することになる。From these facts, it follows that approximately 3 epoxy groups are present per molecule of the heparin derivative synthesized in the present invention.
また、上記作製したヘパリン誘導体中のアミノ基の串を
下記ニンヒドリン法の手順に従い測定し１こ。In addition, the presence of amino groups in the heparin derivative prepared above was measured according to the following procedure of the ninhydrin method.
１）ニンヒドリン２．０ｇ、ヒドリンダンヂン０゜３り
を７５ｍｅのメヂルゼロソルアに溶Ｗ（する。更に４Ｎ
！！ＩＮ酸）−１−リウムバツフ７２５ｄを７１［＋え
、ニンヒドリン試薬とする。1) Dissolve 2.0 g of ninhydrin and 0.3 liters of hydrin danjin in 75me Mejil Zero Solua.
! ! The IN acid)-1-lium buffer 725d is converted into 71[+] and used as a ninhydrin reagent.
２）ヘパリン及び上記作製したヘパリン誘導体１％水溶
液０．７５ｒｄｌを試験官にそれぞれ入れる。2) Place heparin and 0.75 rdl of the 1% aqueous solution of the heparin derivative prepared above into a test tube.
３）更に、１）で作製したニンヒドリン試薬０゜５ｄを
加え、撹拌後１００℃で１５分間反応させる。3) Furthermore, add 0.5 d of the ninhydrin reagent prepared in 1), and after stirring, react at 100° C. for 15 minutes.
４）冷却後５０％エタノール水溶液５ ｎ＆をハ（１え
、直らに５７ Ｑ ｎ ｍで吸光匪を測定する。4) After cooling, add 5 nm of 50% ethanol aqueous solution and immediately measure the absorbance at 57 Q nm.
結果を第１表に示す。The results are shown in Table 1.
第１表
第１表に示すように、ヘパリンが本発明のヘパリン誘４
体となることによって、アミノ基は大部分が消失し、こ
れによりエポキシ基とアミノ基との１７１異的な反応が
明確となった。Table 1 As shown in Table 1, heparin is the heparin inducer of the present invention.
As a result, most of the amino groups disappeared, and the 171 different reaction between the epoxy group and the amino group became clear.
また、ヘパリン誘導体合成反応中のヘパリン活性（Ｓ−
２２２２基質による力（色測定）を９日目まで追跡した
ところ、第２図に示すように６日目までその活性に変化
は認められなかった。ヘパリンの反応朝明ｅ４ｍは３．
７５甲位／ｄであった。In addition, heparin activity (S-
When the activity (color measurement) of the 2222 substrate was followed up to the 9th day, no change was observed in its activity up to the 6th day, as shown in FIG. Heparin reaction morning e4m is 3.
It was 75 ko/d.
更に、セルロースへのヘパリン結合量と反応時間との関
係を調べたところ、第３図に示すように反応４日「１で
セルロースへの最大結合量が認められた。Furthermore, when the relationship between the amount of heparin bound to cellulose and the reaction time was investigated, as shown in FIG. 3, the maximum amount of heparin bound to cellulose was observed on day 4 of the reaction.
参考例１
ヘパリン誘導体とセルロースとの結合処理セルロースシ
ート片（１ｃｍＸ３ｃｍ＞を０．５％水酸化すトリウム
水）容液中に３０分間浸漬した。Reference Example 1 Bonding Treatment of Heparin Derivative and Cellulose A piece of cellulose sheet (1 cm x 3 cm) was immersed in a 0.5% thorium hydroxide solution for 30 minutes.
上記アルカリ５ｉ！！理したセルロース片を取り出し、
セルロース片面上の水酸化すトリウム水溶液をかるく除
去した。同様に得られたセルロース片３片を０．２％ヘ
パリン誘導体水溶液（３７！］、８ＩＵ／−）の１０ｄ
中に浸漬し、所定時間経過後、充分に洗浄し、ヘパリン
結合量を測定した。Alkaline 5i above! ! Take out the processed cellulose pieces,
The thorium hydroxide aqueous solution on one side of the cellulose was slightly removed. Three pieces of cellulose obtained in the same manner were added to 10 d of 0.2% heparin derivative aqueous solution (37!, 8 IU/-).
After a predetermined period of time, the sample was thoroughly washed and the amount of heparin bound was measured.
ヘパリン但測定には以下のＡルト・トルイジンブルー改
良法を用いた。For heparin measurement, the following modified A-toluidine blue method was used.
１）セルロース片を試験管に取り、０．０４％１〜ルイ
ジンブルー液を入れ５分間放置する。1) Take a piece of cellulose in a test tube, add 0.04% 1~Luisine blue solution and leave it for 5 minutes.
２）蒸溜水１０ｒｎｉで２回、デカンテーション洗浄す
る。2) Wash by decantation twice with 10 rni of distilled water.
３）０．ＩＮ塩Ｍ５ｒｎｉを加え、１０分間敢首し、デ
カ−ジョン後、再度同様に０．１Ｎ［酸！Ｍの処理を行
なう。3) 0. Add IN salt M5rni, incubate for 10 minutes, decurse, and add 0.1N [acid!] again in the same manner. Process M.
４）蒸溜水１０ｍを加え、１０分間放置し、デカ−ジョ
ン後、再度同様に蒸溜水１０ｄの処理を行なう。4) Add 10 ml of distilled water, leave to stand for 10 minutes, decurse, and then process 10 d of distilled water again in the same manner.
５）セルロース片表面上の水滴を取り除き、上記セルロ
ース片６枚分を一本の試験管に取る。5) Remove water droplets on the surface of the cellulose pieces, and take 6 cellulose pieces into one test tube.
６）更に、緩衝液（エタノールと０．１Ｎ７に酸化ナト
リウム水溶液４対１容皐比）を入れ、２０〜３０分後に
５３０ｎｍで吸光度を測定する。6) Furthermore, add a buffer solution (ethanol and 0.1N7 to sodium oxide aqueous solution 4:1 volume ratio) and measure the absorbance at 530 nm after 20 to 30 minutes.
この吸光度測定の結果を第１図に示す。第４図に示され
るように、セルロースとヘパリン誘導体は、反応開始よ
り６時間経過後はヘパリン結合量が一定であった。The results of this absorbance measurement are shown in FIG. As shown in FIG. 4, the amount of heparin bound between the cellulose and the heparin derivative was constant after 6 hours from the start of the reaction.
（発明の効果）
以上詳述したにうに、本発明によれば、ヘパリンと２個
以上のエポキシ基をイ１するエポキシ化合物との反応生
成物であって、ヘパリンのアミノ基とエポキシ化合物の
エポキシ基とが結合し、かつ、反応生成物中に未反応の
エポキシ基が残存しているヘパリン誘導体が１７られる
。(Effects of the Invention) As detailed above, according to the present invention, there is provided a reaction product between heparin and an epoxy compound having two or more epoxy groups. A heparin derivative 17 in which the epoxy group is bonded to the epoxy group and an unreacted epoxy group remains in the reaction product is obtained.
このヘパリンｍ４体は、ヘパリンの抗血液凝固活性が失
なわれていない。また、未反１心のエポキシ基を有して
いるので水酸基、アミノ基、カルボキシル基等を有する
高分子と容易に結合し得るため、抗血栓材料の製造にお
ける中聞体として４へめでｎ用である。This heparin m4 body has not lost its anti-coagulant activity. In addition, since it has an epoxy group with an unreflected single core, it can easily bond with polymers having hydroxyl groups, amino groups, carboxyl groups, etc., so it can be used as a medial body in the production of antithrombotic materials. It is.
ざらに本発明による上記誘導体の製造法は、ヘパリンと
２個以上のエポキシ基を有するエポキシ化合物とをＤＨ
９−１２の水溶液中至温ないし１００℃の温度で反応さ
けることからなり、特別の反応装置を必要とけず、ｒａ
ｒ、＋ｉな操作で、反［芯生成物中に未反応のエポキ
シ基が残存し、かつヘパリンの抗血＠凝固作用を保持し
たヘパリン誘導体を装造することができる。In general, the method for producing the above-mentioned derivatives according to the present invention is to combine heparin and an epoxy compound having two or more epoxy groups with DH.
It consists of avoiding the reaction in an aqueous solution of 9-12 at a temperature of from the lowest temperature to 100°C, and there is no need for a special reaction apparatus, and the ra
r, +i operations, it is possible to prepare a heparin derivative in which unreacted epoxy groups remain in the anti-core product and retain the anti-coagulant action of heparin.
第１図は、ヘパリンと１，４−ブタンジオールシグリシ
ジルエーテルとの反応率を示すグラフである。
第２図は、ヘパリン誘導体合成１１．ｉのヘパリンの活
性変化を示すグラフである。
第３図は、セルロースへのヘパリン結合量と反応時間と
の関係を示すグラフである。
第４図は、セルロースとヘパリン、１１体の反応時間と
結合重を示すグラフである。
特許出願人 チル七株式会社
時間旧）
第１図
第２図
第３図FIG. 1 is a graph showing the reaction rate between heparin and 1,4-butanediol siglycidyl ether. Figure 2 shows heparin derivative synthesis 11. It is a graph showing the change in heparin activity of i. FIG. 3 is a graph showing the relationship between the amount of heparin bound to cellulose and reaction time. FIG. 4 is a graph showing the reaction time and bond weight of 11 bodies of cellulose and heparin. Patent applicant: Chiru Seven Co., Ltd.) Figure 1 Figure 2 Figure 3
Claims (6)
シ化合物との反応生成物であって、ヘパリンのアミノ基
とエポキシ化合物のエポキシ基とが結合し、かつ反応生
成物中に未反応のエポキシ基が残存していることを特徴
とするヘパリン誘導体。(1) A reaction product of heparin and an epoxy compound having two or more epoxy groups, in which the amino group of heparin and the epoxy group of the epoxy compound are bonded, and there are unreacted epoxy groups in the reaction product. A heparin derivative characterized by remaining.
を特徴とする特許請求の範囲第１項に記載のヘパリン誘
導体。(2) The heparin derivative according to claim 1, wherein the epoxy compound is a diglycidyl compound.
グリシジルエーテルであることを特徴とする特許請求の
範囲第２項に記載のヘパリン誘導体。(3) The heparin derivative according to claim 2, wherein the diglycidyl compound is 1,4-butanediol diglycidyl ether.
シ化合物との反応生成物であって、ヘパリンのアミノ基
とエポキシ化合物のエポキシ基とが結合し、かつ反応生
成物中に未反応のエポキシ基が残存しているヘパリン誘
導体を製造するにあたり、ペパリンと２個以上のエポキ
シ基を有するエポキシ化合物とをｐＨ７以上の液体中で
反応させることを特徴とするヘパリン誘導体の製造法。(4) A reaction product of heparin and an epoxy compound having two or more epoxy groups, in which the amino group of heparin and the epoxy group of the epoxy compound are bonded, and there are unreacted epoxy groups in the reaction product. 1. A method for producing a heparin derivative, which comprises reacting peparin with an epoxy compound having two or more epoxy groups in a liquid having a pH of 7 or higher.
の製造法。(5) The manufacturing method according to claim 4, wherein the pH is 8 to 10.
する特許請求の範囲第４項記載の製造法。(6) The production method according to claim 4, wherein the reaction is carried out in an aqueous solution having a pH of 9 at room temperature.
Priority Applications (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP28666186A JPS63139901A (en) | 1986-12-03 | 1986-12-03 | Heparin derivative and its preparation |
| DE19873786927 DE3786927T2 (en) | 1986-12-03 | 1987-12-02 | ANTITHROMBOTIC DRUGS AND THEIR PRODUCTION. |
| PCT/JP1987/000934 WO1988004183A1 (en) | 1986-12-03 | 1987-12-02 | Antithrombotic medical materials and process for their production |
| EP19870907978 EP0336964B1 (en) | 1986-12-03 | 1987-12-02 | Antithrombotic medical materials and process for their production |
| DK430188A DK430188A (en) | 1986-12-03 | 1988-08-02 | ANTITHROMOBOLIC MATERIAL FOR MEDICAL USE AND PROCEDURES FOR PRODUCING THEREOF |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP28666186A JPS63139901A (en) | 1986-12-03 | 1986-12-03 | Heparin derivative and its preparation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS63139901A true JPS63139901A (en) | 1988-06-11 |
Family
ID=17707314
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP28666186A Pending JPS63139901A (en) | 1986-12-03 | 1986-12-03 | Heparin derivative and its preparation |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS63139901A (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE3109141A1 (en) * | 1981-03-11 | 1982-09-23 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V., 8000 München | Antithrombogenic treatment of matrix surfaces |
-
1986
- 1986-12-03 JP JP28666186A patent/JPS63139901A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE3109141A1 (en) * | 1981-03-11 | 1982-09-23 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V., 8000 München | Antithrombogenic treatment of matrix surfaces |
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