JP3007785B2 - Thrombomodulin composition and method for preventing denaturation thereof - Google Patents
Thrombomodulin composition and method for preventing denaturation thereofInfo
- Publication number
- JP3007785B2 JP3007785B2 JP6043837A JP4383794A JP3007785B2 JP 3007785 B2 JP3007785 B2 JP 3007785B2 JP 6043837 A JP6043837 A JP 6043837A JP 4383794 A JP4383794 A JP 4383794A JP 3007785 B2 JP3007785 B2 JP 3007785B2
- Authority
- JP
- Japan
- Prior art keywords
- thrombomodulin
- composition
- salts
- arginine
- freeze
- 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.)
- Expired - Lifetime
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Description
【０００１】[0001]
【発明の属する技術分野】本発明は、トロンボモジュリ
ン、およびアミノ酸またはその塩類からなる群から選ば
れた一種または二種以上を含有することを特徴とするト
ロンボモジュリン組成物および該組成物の凍結乾燥工程
でのトロンボモジュリンの変性防止方法に関する。TECHNICAL FIELD The present invention relates to a thrombomodulin composition comprising one or more selected from the group consisting of thrombomodulin and amino acids or salts thereof, and a freeze-drying step of the composition. A method for preventing denaturation of thrombomodulin.
【０００２】[0002]
【従来の技術】現在、血栓溶解剤として用いられている
ものには、ストレプトキナーゼ、ウロキナーゼや組織プ
ラスミノーゲンアクチベーターがある。また、抗血液凝
固剤としてはヘパリンやワーファリンが用いられてい
る。さらに、血小板凝集抑制剤としてはアスピリン、ス
ルフィンピラゾン、ジピリダモール等が使われている。2. Description of the Related Art Streptokinase, urokinase and tissue plasminogen activator are currently used as thrombolytic agents. Heparin and warfarin are used as anticoagulants. Further, as a platelet aggregation inhibitor, aspirin, sulfinpyrazone, dipyridamole and the like are used.
【０００３】これらの血栓溶解剤、抗血液凝固剤および
血小板凝集抑制剤は、それぞれ別個に、あるいは併用し
て、たとえば、心筋梗塞、血栓症、塞栓症、末梢血管閉
塞症、閉塞性動脈硬化症、血管内血液凝固症候群（ＤＩ
Ｃ）、狭心症、一過性脳虚血発作、妊娠中毒症等の疾患
の治療および予防に用いられている。しかしながら、こ
れらの血栓溶解剤、抗血液凝固剤および血小板凝集抑制
剤は非常に複雑な機構から成り立つ血液の凝固線溶系の
ごく一部に作用するにすぎない。そこで、血液の凝固線
溶系に広く作用し、優れた血液凝固抑制作用を示す薬剤
が求められていた。[0003] These thrombolytic agents, anticoagulants and platelet aggregation inhibitors are used separately or in combination, for example, for example, myocardial infarction, thrombosis, embolism, peripheral vascular occlusion, obstructive arteriosclerosis. , Intravascular Coagulation Syndrome (DI
C), for the treatment and prevention of diseases such as angina pectoris, transient cerebral ischemic attacks, and preeclampsia. However, these thrombolytics, anticoagulants and platelet aggregation inhibitors act only on a small part of the blood coagulation / fibrinolysis system, which consists of a very complex mechanism. Thus, there has been a demand for a drug that acts widely on the coagulation / fibrinolysis system of blood and has an excellent blood coagulation inhibitory action.
【０００４】ところで、トロンボモジュリン（以下ＴＭ
と略称する）はトロンビンによるプロテインＣ活性化を
促進する作用を有する。プロテインＣは血液凝固線溶系
において重要な役割を演じているビタミンＫ依存性の蛋
白質であり、トロンビンの作用により活性化される。活
性型プロテインＣは、生体内で血液凝固系因子の活性型
第Ｖ因子、および活性型第ＶＩＩＩ因子を失活させ、ま
た血栓溶解作用を有するプラスミノーゲンアクチベータ
ーの産生に関与していることが知られている［鈴木宏
治、医学のあゆみ、第１２５巻、９０１頁（１９８３
年）］。By the way, thrombomodulin (hereinafter referred to as TM)
Has an action to promote the activation of protein C by thrombin. Protein C is a vitamin K-dependent protein that plays an important role in the blood coagulation / fibrinolysis system and is activated by the action of thrombin. Activated protein C inactivates the activated coagulation system factors V and VIII in vivo and is involved in the production of plasminogen activator having a thrombolytic action [Koji Suzuki, History of Medicine, Vol. 125, p. 901 (1983)
Year)].
【０００５】ＴＭは、このトロンビンによるプロテイン
Ｃの活性化を促進して抗血液凝固作用と血栓溶解作用を
示す活性型プロテインＣを大量に産生せしめるものであ
る。従って、ＴＭは生体における抗血液凝固および血栓
溶解に大きく寄与するものである。前記のように、ＴＭ
は抗血液凝固作用と血小板凝集抑制作用および血栓溶解
作用を有するのでたとえば、心筋梗塞、血栓症、塞栓
症、末梢血管閉塞症、閉塞性動脈硬化症、血管内血液凝
固症候群（ＤＩＣ）、狭心症、一過性脳虚血発作、妊娠
中毒症の患者の治療および予防に用いられることが期待
される。[0005] TM promotes the activation of protein C by thrombin to produce a large amount of active protein C having an anticoagulant effect and a thrombolytic effect. Therefore, TM greatly contributes to anticoagulation and thrombolysis in a living body. As mentioned above, TM
Has an anticoagulant effect, a platelet aggregation inhibitory effect and a thrombolytic effect. It is expected to be used for the treatment and prevention of patients with cerebral illness, transient ischemic attack, preeclampsia.
【０００６】ＴＭは、細胞からの産生量が少ないので、
工業的規模での生産は行われていなかった。しかし、遺
伝子組み換え体の利用（山本ら、特開昭６４−６２１９
号公報）でＴＭを容易に得ることが可能となり、その医
薬品としての開発が行われるようになった。[0006] Since the amount of TM produced from cells is small,
There was no production on an industrial scale. However, the use of recombinants (Yamamoto et al., JP-A-64-6219)
Publication), it has become possible to easily obtain TM, and its development as a drug has come to be carried out.
【０００７】ＴＭを抗血液凝固剤あるいは血栓溶解剤と
して広く安定的に供給するためには凍結乾燥を行って製
剤化することは必須の操作である。ところが、本発明者
らはＴＭ含有溶液を凍結乾燥すると、微量ではあるが一
部が変性によって高分子化し、ＴＭ分子がいくつか会合
した多量体が生成することを明らかにした。蛋白質の凍
結乾燥では水分が一部水和層まで脱水され、部分的に環
境が極端に非水化されることによって、蛋白質の構造保
持機構が破壊され、変性が起こるものと考えられる。Ｔ
Ｍは医薬品として開発されているものであるため、変性
物を含んでいる場合には、その抗原性を含めた安全性が
問題となる。このような実状では、その抗血液凝固作用
や血栓溶解作用にもかかわらず、治療薬として安全なＴ
Ｍ組成物を提供することは不可能である。[0007] In order to widely and stably supply TM as an anticoagulant or thrombolytic agent, it is an essential operation to freeze-dry it to prepare a formulation. However, the present inventors have clarified that when a TM-containing solution is freeze-dried, a small amount of a part thereof is polymerized by denaturation, and a multimer in which some TM molecules are associated is formed. In lyophilization of proteins, it is considered that water is partially dehydrated to a hydrated layer, and the environment is partially dehydrated, thereby destroying the structure retention mechanism of proteins and causing denaturation. T
Since M has been developed as a pharmaceutical, its safety, including its antigenicity, becomes an issue when it contains denatured products. In such a situation, despite its anticoagulant and thrombolytic effects, safe T
It is not possible to provide an M composition.
【０００８】[0008]
【発明が解決しようとする課題】前記のように、変性物
を含むＴＭ組成物は人体に投与することは好ましくな
く、治療薬として用いる場合は変性物を含まないＴＭ組
成物が望ましい。このため、ＴＭの凍結乾燥工程での変
性による高分子化を防止し、安全で安定なＴＭ組成物お
よび該組成物を得る方法が要求される。As described above, it is not preferable to administer a TM composition containing a denatured substance to the human body, and when used as a therapeutic agent, a TM composition containing no denatured substance is desirable. Accordingly, there is a need for a safe and stable TM composition and a method for obtaining the composition, which prevents the TM from being polymerized due to denaturation in the freeze-drying step.
【０００９】[0009]
【課題を解決するための手段】本発明者らは前記の問題
点を解決するために鋭意研究を行った結果、アミノ酸ま
たはその塩類より選択される一種または二種以上を添加
することで、凍結乾燥工程でのＴＭの変性を防止しうる
ことを見い出し、良好なＴＭ凍結乾燥組成物が得られ、
本発明を完成した。Means for Solving the Problems The present inventors have conducted intensive studies to solve the above-mentioned problems, and as a result, by adding one or more selected from amino acids or salts thereof, It has been found that denaturation of TM in the drying step can be prevented, and a good freeze-dried TM composition is obtained.
The present invention has been completed.
【００１０】本発明は、上記の知見に基づいて完成され
てもので、トロンボモジュリンを含み、アルギニン、グ
ルタミン酸、プロリン、セリン、グリセリンおよびその
塩類よりなる群から選ばれた一種また二種以上のアミノ
酸またはその塩類を少なくとも含有することを特徴とす
るトロンボモジュリン組成物、およびトロンボモジュリ
ンを含み、アルギニン、グルタミン酸、プロリン、セリ
ン、グリシンおよびその塩類からなる群から選ばれた一
種または二種以上のアミノ酸またはその塩類を少なくと
も含有することを特徴とする凍結乾燥工程でのトロンボ
モジュリンの変性防止方法である。The present invention has been completed on the basis of the above findings, and includes one or more amino acids or amino acids selected from the group consisting of arginine, glutamic acid, proline, serine, glycerin and salts thereof, including thrombomodulin. A thrombomodulin composition characterized by containing at least its salts, and comprising thrombomodulin, one or more amino acids or salts thereof selected from the group consisting of arginine, glutamic acid, proline, serine, glycine and salts thereof. A method for preventing denaturation of thrombomodulin in a freeze-drying step, characterized in that the method comprises at least containing thrombomodulin.
【００１１】本発明でトロンボモジュリンとは、トロン
ビンによるプロテインＣ活性化を促進する作用を有する
物質として定義され、特に好ましくは可溶性のトロンボ
モジュリンが挙げられる。In the present invention, thrombomodulin is defined as a substance having an action of promoting the activation of protein C by thrombin, and particularly preferably, soluble thrombomodulin is used.
【００１２】まず本発明に用いられるＴＭ原料は公知の
方法、またはそれに準じて調製すればよいが、そのよう
なものとして、例えば、前記山本らの方法（特開昭６４
−６２１９号、実施例参照）が挙げられる。すなわち、
ヒト由来のＴＭ遺伝子を遺伝子操作技術により調製し、
必要に応じた改変を行って組み込んだチャイニーズハム
スター卵巣細胞を培養し、培養液から高純度に生成され
たもので、細胞質ドメインを含まない可溶型ＴＭ、例え
ば光散乱法にて分子量６２，０００の可溶ＴＭが挙げら
れる。なお、本発明に用いられるＴＭ原料は、適宜の糖
鎖を有していても良い。First, the TM raw material used in the present invention may be prepared by a known method or an analogous method. Examples of such a method include the method of Yamamoto et al.
-6219, see Examples). That is,
A human-derived TM gene is prepared by a genetic engineering technique,
The Chinese hamster ovary cells, which have been incorporated after being modified as necessary, are cultured and produced in high purity from the culture solution, and a soluble TM containing no cytoplasmic domain, for example, a molecular weight of 62,000 by a light scattering method. Soluble TM. The TM raw material used in the present invention may have an appropriate sugar chain.
【００１３】また本発明に用いられるＴＭ原料の生産方
法は、これらに限定されるものではない。すなわち、Ｔ
Ｍを生産するような組織またはこれら組織培養液から抽
出精製するような原料生産方法も採用できる。一般には
さらに抽出精製工程を経ることにより、適宜等張化剤、
緩衝化剤（リン酸ナトリウムなど）といった塩類を含む
ＴＭ含有溶液として調製すればよい。The method for producing the TM raw material used in the present invention is not limited to these. That is, T
Materials producing M or a raw material producing method such as extraction and purification from these tissue cultures can also be employed. In general, by further performing an extraction and purification step, an isotonic agent as appropriate,
It may be prepared as a TM-containing solution containing salts such as a buffering agent (such as sodium phosphate).
【００１４】また本発明における凍結乾燥工程でのトロ
ンボモジュリンの変性を防止する添加物としては、アミ
ノ酸およびその塩類からなる群から選ばれた一種または
二種以上の有効量が用いられる。このアミノ酸またはそ
の塩類としては、例えばアルギニン、グルタミン酸、プ
ロリン、セリン、グリシンおよびその塩類よりなる群か
ら選ばれた一種または二種以上が挙げられ、好ましくは
アルギニンまたはその酸付加塩、グルタミン酸またはそ
の塩基付加塩または遊離型としてのプロリン、セリン、
グリシンなどが挙げられる。In the present invention, as an additive for preventing denaturation of thrombomodulin in the freeze-drying step, one or more effective amounts selected from the group consisting of amino acids and salts thereof are used. Examples of the amino acid or a salt thereof include one or more selected from the group consisting of arginine, glutamic acid, proline, serine, glycine and salts thereof, preferably arginine or an acid addition salt thereof, glutamic acid or a base thereof. Proline, serine as an addition salt or free form,
Glycine and the like.
【００１５】上記の添加物において最も好ましくは、ア
ルギニンまたはその酸付加塩、グルタミン酸またはその
塩基付加塩、プロリン、セリンである。アルギニン酸付
加塩の場合、付加し得る酸としては製剤学的に許容され
たものであれば特に制限はなく、たとえば、塩酸、クエ
ン酸、硫酸など、ならびにそれらと機能上同等の物質を
挙げることができる。同様に、グルタミン酸塩基付加塩
の場合は、付加し得る塩基としては製剤学的に許容され
るものであれば特に制限はなく、たとえば、ナトリウ
ム、カリウムなど、ならびにそれらと機能上同等の物質
を挙げることができる。Among the above additives, most preferred are arginine or an acid addition salt thereof, glutamic acid or a base addition salt thereof, proline and serine. In the case of an arginic acid addition salt, the acid that can be added is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include hydrochloric acid, citric acid, sulfuric acid, and the like, and substances functionally equivalent thereto. Can be. Similarly, in the case of a glutamic acid base addition salt, the base that can be added is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include sodium, potassium, and the like, and substances functionally equivalent thereto. be able to.
【００１６】上記添加物を含む本発明の組成物において
は、上記添加物の他に第３成分として、糖類や等張化
剤、緩衝化剤などを添加してもよく、特にこれらの影響
を受けるものではないが、等張化剤や緩衝化剤に用いら
れる塩類、特に塩化ナトリウムの濃度が高いことは本発
明を用いて凍結乾燥を行う際、ケーキの形成に害を及ぼ
すこともある。またケーキ形成補助剤あるいはＴＭの容
器への吸着を防止する目的で、適宜アルブミン、ゼラチ
ン等を添加してもよい。また、糖類としては、例えば単
糖類または二糖類が好適であり、特に好ましくはマンニ
トール、トレハロース、ラクトースまたスクロースであ
る。In the composition of the present invention containing the above additives, a saccharide, a tonicity agent, a buffering agent and the like may be added as a third component in addition to the above additives. Although not subject to the present invention, a high concentration of salts used for the tonicity agent or the buffering agent, particularly sodium chloride, may harm the formation of a cake when freeze-drying is performed using the present invention. Further, albumin, gelatin or the like may be appropriately added for the purpose of preventing adsorption of the cake forming aid or TM to the container. Further, as the saccharide, for example, a monosaccharide or a disaccharide is suitable, and particularly preferably, mannitol, trehalose, lactose or sucrose.
【００１７】本発明で用いる添加物の添加量は、通常Ｔ
Ｍ１ｍｇあたり０．０１〜１ｍｍｏｌが例示される。す
なわち、添加量が０．０１ｍｍｏｌ以上となれば本発明
の好ましい変性防止効果が認められるようになり、ま
た、１ｍｍｏｌを越えて添加しても、変性防止効果を大
きく増大させるものではなく、経済的な面において選択
することができる。The amount of the additive used in the present invention is usually T
An example is 0.01 to 1 mmol per mg of M. That is, if the amount of addition is 0.01 mmol or more, the preferred denaturing prevention effect of the present invention will be recognized, and even if added over 1 mmol, the denaturation preventing effect will not be greatly increased, and economical Can be selected in various aspects.
【００１８】添加方法は特に限定されないが、たとえ
ば、添加物を直接ＴＭ含有溶液に添加する方法、または
あらかじめ添加物を水、注射用蒸留水あるいは適当な緩
衝液に溶解して添加する方法などが挙げられる。添加時
期は凍結乾燥前であれば分離精製過程であっても、製剤
化工程であってもよい。The method of addition is not particularly limited. For example, a method in which the additive is directly added to the TM-containing solution, or a method in which the additive is dissolved in water, distilled water for injection or an appropriate buffer in advance and added. No. The time of addition may be a separation / purification process or a formulation step as long as it is before freeze-drying.
【００１９】また、例えば製剤化工程においては、アン
プルまたはバイアルに、水、注射用、蒸留水あるいは適
当な緩衝液１ｍｌあたり０．０５〜１５ｍｇ、好適には
０．１〜５ｍｇのＴＭおよび上記添加物を含有する溶液
を、例えば０．５〜１０ｍｌ充填し、次いで常法により
凍結乾燥して注射用製剤として調整できる。このような
注射用製剤としては、例えば１日１〜３回投与として
０．０１〜１００ｍｇ含有した凍結乾燥製剤として得れ
ばよい。In the formulation step, for example, 0.05 to 15 mg, preferably 0.1 to 5 mg of TM and 0.1 to 5 mg of TM per ml of water, injection, distilled water or a suitable buffer are added to an ampoule or a vial. The solution containing the substance can be filled, for example, in an amount of 0.5 to 10 ml, and then lyophilized by a conventional method to prepare a preparation for injection. Such an injectable preparation may be obtained, for example, as a lyophilized preparation containing 0.01 to 100 mg as an administration once to three times a day.
【００２０】[0020]
【発明の実施の形態】以下、実施例及び比較例により本
発明を具体的に説明するが、本発明は何らこれらによっ
て限定されるものではない。実施例１注射用蒸留水２ｍ
ｌあたり１ｍｇのＴＭ（ヒト由来のＴＭ遺伝子を遺伝子
操作技術により、調製改変して組み込んだチャイニーズ
ハムスター卵巣細胞を培養して得られた、糖鎖を有する
光乱射法にて分子量約６２，０００の可溶型ＴＭ；特開
昭６４−６２１９号）を含む溶液を調製した。このＴＭ
溶液にグルタミン酸ナトリウム０．０５ｍｍｏｌを添加
した。さらに、該溶液を２ｍｌずつガラスバイアル瓶に
分注し、凍結乾燥を行った。凍結乾燥は−４０℃で１８
時間予備凍結し、−４０〜＋２０℃、真空度０．０５〜
０．０１ｍｍＨｇで４０時間一次乾燥し、ついで２０
℃、真空度０．０５〜０．０１ｍｍＨｇで６時間二次乾
燥した。Hereinafter, the present invention will be described in detail with reference to Examples and Comparative Examples, but the present invention is not limited thereto. Example 1 2m of distilled water for injection
1 mg per liter of TM (human TG gene prepared by culturing and incorporating a human-derived TM gene by a genetic engineering technique and having a molecular weight of about 62,000 A solution containing a soluble TM (JP-A-64-6219) was prepared. This TM
To the solution was added 0.05 mmol of sodium glutamate. Further, 2 ml of the solution was dispensed into glass vials and freeze-dried. Lyophilize at -40 ° C for 18
Pre-frozen for -40 to + 20 ° C, degree of vacuum 0.05 to
Primary drying at 0.01 mmHg for 40 hours, followed by 20
Secondary drying was performed at a degree of vacuum of 0.05 to 0.01 mmHg for 6 hours.
【００２１】次いで、この凍結乾燥品を再溶解後、サイ
ズ排除クロマトグラフィー分析を行い高分子化したＴＭ
の割合を求めた。分析には、内径７．５ｍｍ、長さ６０
ｃｍのステンレス管に排除限界分子量５０万の親水性シ
リカゲルを充填したカラムを使用し、０．１Ｍ硫酸ナト
リウムを含む５０ｍＭリン酸ナトリウム緩衝液（ｐＨ
７．０）を溶離液として、波長２１０ｎｍで検出した。
結果は表１に示した。Next, the lyophilized product was redissolved and subjected to size exclusion chromatography analysis to obtain a polymerized TM.
Was determined. For analysis, inner diameter 7.5 mm, length 60
Using a column filled with a hydrophilic silica gel having an exclusion limit molecular weight of 500,000 in a stainless steel tube of 50 cm, a 50 mM sodium phosphate buffer solution (pH: 0.1 M) containing sodium sulfate was used.
7.0) was used as an eluent and detected at a wavelength of 210 nm.
The results are shown in Table 1.
【００２２】[0022]
【表１】 [Table 1]
【００２３】実施例 ２ 添加物をプロリンとした以外は前記の実施例１と同様に
行った。結果は表１に示した。Example 2 The same procedure as in Example 1 was carried out except that the additive was proline. The results are shown in Table 1.
【００２４】実施例 ３ 添加物をセリンとした以外は前記の実施例１と同様に行
った。結果は表１に示した。Example 3 The same procedure as in Example 1 was carried out except that serine was used as an additive. The results are shown in Table 1.
【００２５】実施例 ４ 添加物をグリシンとした以外は前記の実施例１と同様に
行った。結果は表１に示した。Example 4 The same procedure as in Example 1 was carried out except that glycine was used as an additive. The results are shown in Table 1.
【００２６】実施例 ５〜１０ 添加物をアルギニン−塩酸塩とし、添加量を０．０１、
０．０２、０．０５、０．１、０．２、０．５ｍｍｏｌ
とした以外は実施例１と同様に行った。結果は表１に示
した。Examples 5 to 10 The additive was arginine-hydrochloride, and the additive amount was 0.01,
0.02, 0.05, 0.1, 0.2, 0.5 mmol
The procedure was performed in the same manner as in Example 1, except that The results are shown in Table 1.
【００２７】実施例 １１ 添加物として、アルギニン−塩酸塩およびスクロースの
二種を併用し、その添加量をそれぞれ０．０５ｍｍｏｌ
とした以外は前記の実施例１と同様に行った。Example 11 Two kinds of additives, arginine-hydrochloride and sucrose, were used in combination, and the amount of each added was 0.05 mmol.
The procedure was performed in the same manner as in Example 1 except for the above.
【００２８】実施例 １２ 添加物として、アルギニン−塩酸塩０．０５ｍｍｏｌお
よび精製ゼラチン１０ｍｇとした以外は前記の実施例１
と同様に行った。Example 12 The same procedures as in Example 1 were carried out except that 0.05 mmol of arginine-hydrochloride and 10 mg of purified gelatin were used as additives.
The same was done.
【００２９】比較例 １ 添加物を加えなかったこと以外は前記の実施例１と同様
に行った。結果は表２に示した。Comparative Example 1 The procedure of Example 1 was repeated except that no additive was added. The results are shown in Table 2.
【００３０】[0030]
【表２】 [Table 2]
【００３１】比較例 ２〜５ 添加物を塩化ナトリウムとし、添加量を０．０１、０．
０２、０．０５、０．１ｍｍｏｌとした以外は前記の実
施例１と同様に行った。結果は表２に示した。Comparative Examples 2 to 5 The additive was sodium chloride, and the amount of addition was 0.01, 0.
The same procedure was performed as in Example 1 except that the amounts were 02, 0.05, and 0.1 mmol. The results are shown in Table 2.
【００３２】[0032]
【発明の効果】前記の実施例および比較例の結果から明
らかなように、本発明によれば、ＴＭの凍結乾燥工程で
の変性による高分子化を防止し、変性物を含まないＴＭ
組成物を得ることが可能となる。これによって、凝固線
溶系に広く作用し、優れた血液凝固抑制作用を有するＴ
Ｍを安全な治療薬として供給することが可能となる。As is apparent from the results of the above Examples and Comparative Examples, according to the present invention, TM is prevented from being polymerized due to denaturation in the freeze-drying step, and TM containing no denatured product is contained.
It is possible to obtain a composition. As a result, T acts widely on the coagulation / fibrinolysis system and has an excellent blood coagulation inhibitory action.
M can be supplied as a safe therapeutic agent.
【図１】アルギニン−塩酸塩の各種添加量に対するＴＭ
の変性物割合の関係を示す図である。FIG. 1: TM for various amounts of arginine-hydrochloride added
FIG. 4 is a view showing a relationship between denatured product ratios.
Claims (9)
ン、グルタミン酸、プロリン、セリン、グリシンおよび
その塩類よりなる群から選ばれた一種または二種以上の
アミノ酸またはその塩類を少なくとも含有することを特
徴とするトロンボモジュリン組成物。An arginine comprising thrombomodulin.
, Glutamic acid, proline, serine, glycine and
A thrombomodulin composition comprising at least one or two or more amino acids selected from the group consisting of salts thereof or salts thereof.
またはその塩類である請求項１に記載のトロンボモジュ
リン組成物。2. The thrombomodulin composition according to claim 1, wherein the amino acid or a salt thereof is arginine or a salt thereof.
ジュリン１ｍｇあたり０．０１〜１ｍｍｏｌの添加量で
ある請求項１又は２に記載のトロンボモジュリン組成
物。Wherein the amino acid or its salts, thrombomodulin composition according to claim 1 or 2 which is added the amount of bets Ronbomo <br/> forest 1mg per 0.01 to 1 mmol.
１〜３のいずれかに記載のトロンボモジュリン組成物。4. A composition thrombomodulin composition according to any one of claims 1 to 3, which is a freeze-dried composition.
ボモジュリンである請求項１〜４のいずれかに記載のト
ロンボモジュリン組成物。5. thrombomodulin, thrombomodulin composition according to any one of claims 1 to 4 which is a soluble thrombomodulin.
ン、グルタミン酸、プロリン、セリン、グリシンおよび
その塩類からなる群から選ばれた一種または二種以上の
アミノ酸またはその塩を少なくとも含有することを特徴
とする凍結乾燥工程でのトロンボモジュリンの変性防止
方法。6. A freeze-drying step comprising thrombomodulin and at least one or more amino acids or salts thereof selected from the group consisting of arginine, glutamic acid, proline, serine, glycine and salts thereof. For preventing thrombomodulin from denaturing.
またはその塩類である請求項６に記載のトロンボモジュ
リンの変性防止方法。7. The method for preventing denaturation of thrombomodulin according to claim 6, wherein the amino acid or a salt thereof is arginine or a salt thereof.
ジュリン１ｍｇあたり０．０１〜１ｍｍｏｌの添加量で
ある請求項６又は７に記載のトロンボモジュリンの変性
防止方法。8. amino acids and their salts, preparative Ronbomo <br/> modified method for preventing thrombomodulin according to claim 6 or 7 is the addition amount of 0.01~1mmol per forest 1 mg.
ボモジュリンである請求項６〜８のいずれかに記載のト
ロンボモジュリンの変性防止方法。9. The method for preventing denaturation of thrombomodulin according to claim 6 , wherein the thrombomodulin is soluble thrombomodulin.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP6043837A JP3007785B2 (en) | 1993-03-16 | 1994-03-15 | Thrombomodulin composition and method for preventing denaturation thereof |
| JP10280707A JPH11171790A (en) | 1994-03-15 | 1998-10-02 | Denaturation preventing agent for thrombomodulin |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP5564293 | 1993-03-16 | ||
| JP5-55642 | 1993-03-16 | ||
| JP6043837A JP3007785B2 (en) | 1993-03-16 | 1994-03-15 | Thrombomodulin composition and method for preventing denaturation thereof |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP10280707A Division JPH11171790A (en) | 1994-03-15 | 1998-10-02 | Denaturation preventing agent for thrombomodulin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH06321805A JPH06321805A (en) | 1994-11-22 |
| JP3007785B2 true JP3007785B2 (en) | 2000-02-07 |
Family
ID=26383673
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP6043837A Expired - Lifetime JP3007785B2 (en) | 1993-03-16 | 1994-03-15 | Thrombomodulin composition and method for preventing denaturation thereof |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP3007785B2 (en) |
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|---|---|---|---|---|
| EP0689843B1 (en) * | 1993-12-17 | 2003-09-10 | Mochida Pharmaceutical Co., Ltd. | Composition containing soluble thrombomodulins |
| US6551794B1 (en) * | 1995-11-09 | 2003-04-22 | E. R. Squibb & Sons, Inc. | Stable biotinylated biomolecule composition |
| US5770700A (en) * | 1996-01-25 | 1998-06-23 | Genetics Institute, Inc. | Liquid factor IX formulations |
| WO1999018994A1 (en) | 1997-10-15 | 1999-04-22 | Asahi Kasei Kogyo Kabushiki Kaisha | Method for keeping the quality of aqueous parenteral solution of thrombomodulin in storage and distribution |
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| WO2003061687A1 (en) * | 2002-01-18 | 2003-07-31 | Asahi Kasei Pharma Corporation | High-concentration preparation of soluble thrombomodulin |
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- 1994-03-15 JP JP6043837A patent/JP3007785B2/en not_active Expired - Lifetime
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8641925B2 (en) | 2003-07-15 | 2014-02-04 | Nof Metal Coatings Europe | Use of yttrium, zirconium, lanthanum, cerium, praseodymium and/or neodymium as reinforcing agent for an anticorrosion coating composition |
| US8952137B2 (en) | 2007-03-23 | 2015-02-10 | Asahi Kasei Pharma Corporation | Method for producing high-purity soluble thrombomodulin |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH06321805A (en) | 1994-11-22 |
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