JPH0296536A - Dry plasminogen pharmaceutical - Google Patents
Dry plasminogen pharmaceuticalInfo
- Publication number
- JPH0296536A JPH0296536A JP63245485A JP24548588A JPH0296536A JP H0296536 A JPH0296536 A JP H0296536A JP 63245485 A JP63245485 A JP 63245485A JP 24548588 A JP24548588 A JP 24548588A JP H0296536 A JPH0296536 A JP H0296536A
- Authority
- JP
- Japan
- Prior art keywords
- plasminogen
- dry
- albumin
- vol
- preparation
- 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
- 102000013566 Plasminogen Human genes 0.000 title claims abstract description 46
- 108010051456 Plasminogen Proteins 0.000 title claims abstract description 46
- 150000001413 amino acids Chemical class 0.000 claims abstract description 17
- 108010088751 Albumins Proteins 0.000 claims abstract description 15
- 102000009027 Albumins Human genes 0.000 claims abstract description 15
- 150000001720 carbohydrates Chemical class 0.000 claims abstract description 5
- 238000002360 preparation method Methods 0.000 claims description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 12
- 238000004108 freeze drying Methods 0.000 abstract description 6
- 239000000126 substance Substances 0.000 abstract description 5
- 150000005846 sugar alcohols Chemical class 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 239000004475 Arginine Substances 0.000 abstract description 3
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 abstract description 3
- 229930195725 Mannitol Natural products 0.000 abstract description 3
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 abstract description 3
- 239000000594 mannitol Substances 0.000 abstract description 3
- 235000010355 mannitol Nutrition 0.000 abstract description 3
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 abstract description 2
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 abstract description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 abstract description 2
- 239000004472 Lysine Substances 0.000 abstract description 2
- 239000000600 sorbitol Substances 0.000 abstract description 2
- 239000004615 ingredient Substances 0.000 abstract 5
- 230000015572 biosynthetic process Effects 0.000 abstract 1
- 230000002459 sustained effect Effects 0.000 abstract 1
- 235000001014 amino acid Nutrition 0.000 description 13
- 238000000034 method Methods 0.000 description 11
- 239000000243 solution Substances 0.000 description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 7
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 241000700605 Viruses Species 0.000 description 5
- 239000002904 solvent Substances 0.000 description 5
- 238000011282 treatment Methods 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 4
- 239000004471 Glycine Substances 0.000 description 3
- 208000007536 Thrombosis Diseases 0.000 description 3
- 238000004090 dissolution Methods 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000005194 fractionation Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 235000000346 sugar Nutrition 0.000 description 3
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Chemical compound OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- 229920002684 Sepharose Polymers 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 239000003527 fibrinolytic agent Substances 0.000 description 2
- 230000003480 fibrinolytic effect Effects 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 230000002779 inactivation Effects 0.000 description 2
- 210000002826 placenta Anatomy 0.000 description 2
- 210000002381 plasma Anatomy 0.000 description 2
- 229940012957 plasmin Drugs 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 229940124597 therapeutic agent Drugs 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 108010039627 Aprotinin Proteins 0.000 description 1
- 206010003445 Ascites Diseases 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 102000008100 Human Serum Albumin Human genes 0.000 description 1
- 108091006905 Human Serum Albumin Proteins 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 102000003990 Urokinase-type plasminogen activator Human genes 0.000 description 1
- 108090000435 Urokinase-type plasminogen activator Proteins 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- 229960002684 aminocaproic acid Drugs 0.000 description 1
- 229960004405 aprotinin Drugs 0.000 description 1
- 239000003125 aqueous solvent Substances 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical group NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 230000020764 fibrinolysis Effects 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- ZPNFWUPYTFPOJU-LPYSRVMUSA-N iniprol Chemical compound C([C@H]1C(=O)NCC(=O)NCC(=O)N[C@H]2CSSC[C@H]3C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(N[C@H](C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC=4C=CC=CC=4)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC=4C=CC=CC=4)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC2=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC=2C=CC=CC=2)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H]2N(CCC2)C(=O)[C@@H](N)CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N2[C@@H](CCC2)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N2[C@@H](CCC2)C(=O)N3)C(=O)NCC(=O)NCC(=O)N[C@@H](C)C(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@H](C(=O)N1)C(C)C)[C@@H](C)O)[C@@H](C)CC)=O)[C@@H](C)CC)C1=CC=C(O)C=C1 ZPNFWUPYTFPOJU-LPYSRVMUSA-N 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000012931 lyophilized formulation Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 150000004043 trisaccharides Chemical class 0.000 description 1
- 229960005356 urokinase Drugs 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Abstract
Description
【発明の詳細な説明】
［産業上の利用分野］
本発明は、プラスミノゲン製剤の改良に関し、さらに詳
しくは、プラスミノゲン乾燥製剤の水への溶解時の溶解
性の改良をなしたプラスミノゲン乾燥製剤に関するもの
である。Detailed Description of the Invention [Industrial Application Field] The present invention relates to improvement of plasminogen preparations, and more particularly to a plasminogen dry preparation that has improved solubility when dissolved in water. It is.
プラスミノゲンは、ウロキナーゼなどによって活性化さ
れてプラスミノゲンとなり、これがフィブリンを溶解し
て線溶現象をもたらすものである。Plasminogen is activated by urokinase and the like to become plasminogen, which dissolves fibrin and causes fibrinolysis.
プラスミンは、線溶現象の研究用として用いられる他、
近年線溶系の治療剤（血栓治療剤）としての臨床応用も
なされている。Plasmin is used to study fibrinolytic phenomena, and
In recent years, it has also been clinically applied as a fibrinolytic therapeutic agent (thrombus therapeutic agent).
プラスミノゲン製剤において、プラスミノゲン自体は前
駆型酵素で安定な物質であり、夾雑物として製剤中にプ
ラスミン等を含まないかぎり安定である。しかし、プラ
スミノゲンは、乾燥製剤とすることにより、水に対する
溶解性および溶状が悪くなる。In plasminogen preparations, plasminogen itself is a precursor enzyme and is a stable substance, and is stable unless the preparation contains plasmin or the like as a contaminant. However, when plasminogen is made into a dry preparation, its solubility and solubility in water deteriorates.
出願人は先に、プラスミノゲンの乾燥製剤に、水への溶
解時のにごり、糸状様物質の発生を抑止するための熔解
剤として糖またはアミノ酸を添加したプラスミノゲン製
剤を提案している（特開昭６２−１５’３２２４公報）
。The applicant has previously proposed a plasminogen preparation in which sugar or amino acids are added to a dry preparation of plasminogen as a dissolving agent to suppress the generation of cloudiness and filamentous substances when dissolved in water (Japanese Patent Application Laid-open No. 62-15'3224)
.
しかし、プラスミノゲン含有組成物を過酷条件下（たと
えばウィルス不活化のための処理等）におくことにより
、乾燥製剤の溶解性および溶状が悪くなることが判明し
た。However, it has been found that by subjecting the plasminogen-containing composition to harsh conditions (eg, treatment for virus inactivation, etc.), the solubility and solubility of the dry preparation deteriorate.
本発明の目的は、過酷条件下においた後でも乾燥化され
たプラスミノゲンの水に対する溶解性および溶状が良好
であるプラスミノゲン乾燥製剤を提供することである。An object of the present invention is to provide a dried plasminogen preparation in which the dried plasminogen has good solubility and solubility in water even after being subjected to harsh conditions.
本発明はプラスミノゲン、Ｉ！類、アミノ酸およびアル
ブミンを含有してなるプラスミノゲン乾燥製剤からなる
。The present invention provides plasminogen, I! It consists of a dry plasminogen preparation containing amino acids, amino acids, and albumin.
本発明におけるプラスミノゲン乾燥製剤とは、実質的に
乾燥された状態にある製剤をいい、たとえば粉末状とな
しうる程度であれば、多少水分を含んでいてもよい。The dry plasminogen preparation in the present invention refers to a preparation in a substantially dry state, and may contain some moisture, for example, as long as it can be made into a powder.
本発明のプラスミノゲン乾燥製剤は乾燥状プラスミノゲ
ンを主成分とするものである、乾燥状プラスミノゲンと
しては、粉末状、特に凍結乾燥プラスミノゲンがあげら
れる。かかる乾燥状プラスミノゲンは、プラスミノゲン
含有液を、自体既知の手段にて乾燥、特に凍結乾燥する
ことによって製造される。The dried plasminogen preparation of the present invention is mainly composed of dried plasminogen. Examples of the dried plasminogen include powdered plasminogen, particularly lyophilized plasminogen. Such dry plasminogen is produced by drying a plasminogen-containing liquid by a method known per se, particularly by freeze-drying it.
当該プラスミノゲン含有液は、特に限定されるものでは
なく、たとえば人を含む動物等の直情、血漿、腹水、胎
盤抽出液、胎盤＆ｌＩ＃ｓ抽出液、コーンの低温アルコ
ール分画法の百分■のようにプラスミノゲンを含有する
両分から広く公知の方法によって精製されたプラスミノ
ゲン、遺伝子工学により得られたプラスミノゲンなどを
含有する溶液が挙げられる。The plasminogen-containing liquid is not particularly limited, and includes, for example, the blood plasma of animals including humans, plasma, ascites, placenta extract, placenta &lI#s extract, and the 100% fraction of Cohn's low-temperature alcohol fractionation method. Examples include solutions containing plasminogen purified by widely known methods from both components containing plasminogen, plasminogen obtained by genetic engineering, and the like.
プラスミノゲンの精製法としては、たとえば特開昭５５
−１５３５９２号公報に記載の方法、リジン−セファロ
ースによる精製法（Ｓｃｉｅｎｃｅ、 １７０＋１０９
５、１９７０年）などを代表的な方法として例示するこ
とができる。As a method for purifying plasminogen, for example, JP-A-55
-153592, purification method using lysine-Sepharose (Science, 170+109
5, 1970) can be exemplified as a typical method.
また、プラスミノゲン製剤はウィルス不活化のための処
理が行われているものが好ましい。ウィルス不活化処理
としては、たとえばプラスミノゲンの液状組成物を５０
〜１００°Ｃで５〜３０時間加熱する方法、乾燥組成物
を５０〜１００°Ｃで、１０〜１５０時間加熱する方法
、界面活性剤と接触させる方法、その他の薬剤、たとえ
ば、トリアルキルホスフェートと接触させる方法および
これら処理の組み合わせが挙げられる。Furthermore, it is preferable that the plasminogen preparation has been treated to inactivate the virus. For virus inactivation treatment, for example, 50% of a liquid composition of plasminogen is used.
heating the dry composition at 50-100°C for 10-150 hours; contacting with a surfactant; and other agents, e.g. trialkyl phosphate. Examples include contacting methods and combinations of these treatments.
本発明で使用される溶解剤は糖類、アミノ酸およびアル
ブミンである。The solubilizing agents used in the present invention are sugars, amino acids and albumin.
Ｉ！類としては単＄１！類（ブドウ糖、ガラクトースな
ど）、三糖類（シｇ糖、乳零唐など）、糖アルコール（
マンニトール、ソルビトールなど）が例示され、特に好
ましい糖類は糖アルコールである。I! The price is just $1! (glucose, galactose, etc.), trisaccharides (sig sugar, milk sugar, etc.), sugar alcohols (
(mannitol, sorbitol, etc.), and particularly preferred saccharides are sugar alcohols.
アミノ酸としては、中性アミノ酸（グリシン、アラニン
など）、酸性アミノ酸（グルタミン酸、アスパラギン酸
など）、塩基性アミノ酸（アルギニン、リジンなど）が
例示され、特に好ましいアミノ酸は塩基性アミノ酸であ
る。Examples of amino acids include neutral amino acids (glycine, alanine, etc.), acidic amino acids (glutamic acid, aspartic acid, etc.), and basic amino acids (arginine, lysine, etc.), and particularly preferred amino acids are basic amino acids.
アルブミンは、抗原性の問題からヒト由来のアルブミン
であることが好ましく、それらは医療用に精製されたも
のであれば特に制限はない。その純度は、電気泳動法で
分析して８０％以上がアルブミンであるものが好ましい
。ヒト由来アルブミンを得る方法としては、エタノール
分画法（特公昭４７−２８６９号公報、特公昭３５−５
２９７号公報）、有機酸の存在下で加熱する方法（特公
昭４３−１６０４号公報、特公昭５１ ４０ｉ３２１号
公報）なとが例示される。特に、好ましくはアルブミン
を加熱処理（好ましくは、６０゛Ｃ１１０時間程度の加
熱処理）して肝炎ウィルスなどのウィルス不活性化処理
を行ったものが使用される。Albumin is preferably human-derived albumin from the viewpoint of antigenicity, and there are no particular limitations on albumin as long as it is purified for medical use. The purity is preferably 80% or more albumin when analyzed by electrophoresis. As a method for obtaining human-derived albumin, ethanol fractionation method (Japanese Patent Publication No. 47-2869, Japanese Patent Publication No. 35-5
Examples include methods of heating in the presence of an organic acid (Japanese Patent Publication No. 43-1604, Japanese Patent Publication No. 51-40i321). In particular, it is preferable to use albumin that has been heat-treated (preferably heat-treated at 60° C. for about 110 hours) to inactivate viruses such as hepatitis virus.
本発明のより好ましい実施態様は、糖アルコール、塩基
性アミノ酸およびアルブミンの併用である。A more preferred embodiment of the present invention is the combination of sugar alcohol, basic amino acid and albumin.
本発明のプラスミノゲン乾燥製剤は、通常熔解剤をプラ
スミノゲン溶液に添加した後、自体既知の乾燥処理に付
すことによって調製される。The dry plasminogen preparation of the present invention is usually prepared by adding a solubilizing agent to a plasminogen solution and then subjecting it to a known drying treatment.
本発明において、溶解剤の配合量は本発明のプラスミノ
ゲン乾燥製剤を水に熔解させた場合に溶解時のにごり、
糸状様物質の発生を抑制するに十分な量であり、具体的
には各成分毎に次の通りである。In the present invention, the blending amount of the solubilizer is such that when the dry plasminogen preparation of the present invention is dissolved in water, it becomes cloudy at the time of dissolution,
The amount is sufficient to suppress the generation of filamentous substances, and the specific details for each component are as follows.
Ｉ！頻およびアミノ酸の使用量は、それぞれプラスミノ
ゲンの１００〜５００ＣＵ／ｍｌに対して、０．５％（
Ｗ／Ｖ） 〜１０％ （Ｗ／Ｖ）程度、ヨリ好ましくは
０．５〜５％（Ｗ／Ｖ）程度である。なお、糖類〔特に
、糖アルコール〕 （Ａ）と塩基性アミノ酸（Ｂ）との
好ましい重量比、即ちＢ／Ａは０．１〜３であり、最適
には１である。I! The amount of plasminogen and amino acid used is 0.5% (100-500CU/ml of plasminogen).
W/V) to about 10% (W/V), preferably about 0.5 to 5% (W/V). In addition, the preferable weight ratio of saccharide [especially sugar alcohol] (A) and basic amino acid (B), ie, B/A, is 0.1-3, and optimally is 1.
アルブミンの使用量はプラスミノゲンの１００〜５００
ＣＵ／ｍｌに対して０．０５〜５％（Ｗ／Ｖ）程度、
より好ましくは０．５〜１％（Ｗ／Ｖ）程度である。The amount of albumin used is 100 to 500 of that of plasminogen.
About 0.05 to 5% (W/V) to CU/ml,
More preferably, it is about 0.5 to 1% (W/V).
上記の溶解剤の添加量の範囲において、製剤の安定性、
水溶解性、溶状および製剤化のバランスが最も良好であ
る。Within the range of the amount of solubilizer added above, the stability of the formulation,
It has the best balance of water solubility, solubility, and formulation.
本発明のプラスミノゲン乾燥製剤は、用時生理食塩液注
射用蒸留水などの水性溶媒に溶解されて投与される。投
与に際しては、−船釣には静脈内投与が行われ、たとえ
ば血栓症の治療のためには一回当たりブラスミノゲンと
して１００〜３０００ ＣＵの投与が行われる。The dry plasminogen preparation of the present invention is administered after being dissolved in an aqueous solvent such as distilled water for physiological saline injection before use. For administration, intravenous administration is carried out, for example, for the treatment of thrombosis, 100 to 3000 CU of blasminogen is administered per dose.
なお、本発明からなる製剤には、医薬品製造の通例技術
に準じて賦形剤などを有効量添加してもよい。Note that an effective amount of excipients and the like may be added to the preparation of the present invention according to conventional techniques for manufacturing pharmaceuticals.
本発明からなるプラスミノゲン乾燥製剤は、水への溶解
時のにごり、糸状用物質の発生が抑制され、溶解後も活
性が保持される。また、凍結乾燥時の安定性、特に長期
安定性に優れているので、医療用製剤としてより安全で
あり、実用に即したものである。従って、本発明製剤に
よって、たとえば各種血栓症の治療がより効果的に行え
る。The dry plasminogen preparation of the present invention suppresses the generation of cloudiness and filamentous substances when dissolved in water, and retains its activity even after dissolution. Furthermore, since it has excellent stability during freeze-drying, especially long-term stability, it is safer as a medical preparation and is suitable for practical use. Therefore, the preparations of the present invention can more effectively treat, for example, various thromboses.
〔実施例］
実施例１
コーンの冷エタノール分画性で得られた両分■十■ペー
スト抽出残査を、１０単位／ ｍ ｌのアプロチニンと
０．１ Ｍ塩化ナトリウムを含むトリス塩酸緩衝液（ｐ
ｌ（８，３）に懸濁し、少時撹拌した後５％硫酸アンモ
ニウムを添加・撹拌し遠心分離により上清を分離した。[Example] Example 1 The paste extraction residue obtained from the cold ethanol fractionation of Cohn was mixed with Tris-HCl buffer (containing 10 units/ml of aprotinin and 0.1 M sodium chloride). p
After stirring briefly, 5% ammonium sulfate was added and stirred, and the supernatant was separated by centrifugation.
更に、この上清に３０％硫酸アンモニウムを添加・撹拌
し遠心分離により沈澱を分離した。この沈澱を、０．９
％塩化ナトリウムを含む０．９％グリシンン容液（ｐＨ
７，２）に懸濁し、Ｄｅｕｔｓｃｈ、 Ｄ、Ｇら（Ｓｃ
ｉｅｎｃｅ、 １７０．１０９５． （１９７０）
）の方法に準じリジン−セファロースカラムに注入し、
プラスミノゲンを吸着させ、次いで不純蛋白質を１塩化
ナトリウムを含む０．９％グリシン溶液（ｐＨ７，２）
で洗浄した後、０．２Ｍε−アミノカプロン酸と０．９
％塩化ナトリウムとを含む溶媒（ｐＨ７，０）を用いて
吸着したプラスミノゲンを溶出せしめた。Furthermore, 30% ammonium sulfate was added to this supernatant, the mixture was stirred, and the precipitate was separated by centrifugation. This precipitate is 0.9
0.9% glycine solution containing % sodium chloride (pH
7,2) and Deutsch, D, G et al. (Sc
ience, 170.1095. (1970)
), inject into a lysine-Sepharose column, and
Plasminogen is adsorbed, and then impure proteins are removed using a 0.9% glycine solution containing sodium monochloride (pH 7.2).
After washing with 0.2 Mε-aminocaproic acid and 0.9
The adsorbed plasminogen was eluted using a solvent (pH 7.0) containing % sodium chloride.
この精製した精製プラスミノゲン液（７６，２ｃｕ／ｍ
ｌ）をｐＨ５，ｏで６０°ＣＩＯ時間加熱後、１％アル
ギニン・１％マンニトールと０．４５％塩化ナトリウム
を含む５０１リン酸緩衝液（ｐｌ＋７．２ ）で−昼夜
透析した。透析後、最終濃度が０〜５％となるようにヒ
ト血清アルブミンを添加し凍結乾燥した。凍結乾燥後、
蒸留水で再溶解し性状を比較した。This purified plasminogen solution (76.2 cu/m
1) was heated at pH 5.0 for 60°CIO hours, and then dialyzed day and night against 501 phosphate buffer (pl+7.2) containing 1% arginine, 1% mannitol and 0.45% sodium chloride. After dialysis, human serum albumin was added to the final concentration of 0 to 5% and freeze-dried. After freeze-drying,
It was redissolved in distilled water and its properties were compared.
当該凍結乾燥製剤は、容易に蒸留水に溶解した。The lyophilized formulation was easily dissolved in distilled water.
また、溶状は表１に示す通りであり、アルブミンを添加
した再溶解液は清澄であった。Further, the solution state was as shown in Table 1, and the redissolved solution to which albumin was added was clear.
また、アルブミン無添加、０．５％添加の溶解液を室温
で静置して、２時間までの活性および溶状を測定し、表
２に示した。In addition, a solution containing no albumin and 0.5% addition was allowed to stand at room temperature, and the activity and solubility were measured for up to 2 hours, and the results are shown in Table 2.
（以下余白）
表１ 凍結乾燥後再溶解液の性状
＋＋：極めて清澄、＋：清澄、±：＠白濁なお、プラス
ミノゲンの力価は、Ｋａｔｏら（Ｊ。(Margin below) Table 1 Properties of redissolved solution after freeze-drying ++: Extremely clear, +: Clear, ±: @ Cloudy Note that the titer of plasminogen was determined by Kato et al. (J.
Ｂｉｏｃｈｅｍ、 ８８．１８３． （１９８０））の
方法に準じ、合成基質法で求めた。Biochem, 88.183. (1980)), it was determined by the synthetic substrate method.
また、ＣＯ単位はカゼイン単位（Ｖｏｘ Ｓａｎｇ、
５３５７−３７６ （１９６０））を意味する。In addition, the CO unit is a casein unit (Vox Sang,
5357-376 (1960)).
表２ 凍結乾燥後の水熔解液の安定性 率溶解後の室温静置時間 手続補正書（自制 昭和６３年１１月１６日 曾Table 2 Stability of water solution after freeze-drying Room temperature standing time after rate dissolution Procedural amendment (self-restraint) November 16, 1988 so
Claims (1)
有してなることを特徴とするプラスミノゲン乾燥製剤。A dry plasminogen preparation containing plasminogen, saccharides, amino acids, and albumin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63245485A JPH0296536A (en) | 1988-09-29 | 1988-09-29 | Dry plasminogen pharmaceutical |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63245485A JPH0296536A (en) | 1988-09-29 | 1988-09-29 | Dry plasminogen pharmaceutical |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH0296536A true JPH0296536A (en) | 1990-04-09 |
Family
ID=17134362
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63245485A Pending JPH0296536A (en) | 1988-09-29 | 1988-09-29 | Dry plasminogen pharmaceutical |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0296536A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1994015631A1 (en) * | 1993-01-11 | 1994-07-21 | The Green Cross Corporation | Dry plasminogen preparation |
| FR2719479A1 (en) * | 1994-05-04 | 1995-11-10 | Sanofi Elf | Lyophilized stable formulation comprising a protein: assay kit. |
| FR2740686A1 (en) * | 1995-11-03 | 1997-05-09 | Sanofi Sa | STABLE LYOPHILIZED PHARMACEUTICAL FORMULATION |
| US7172999B2 (en) | 1995-10-25 | 2007-02-06 | Roche Diagnostics Gmbh | Method and preparations for stabilizing biological materials by drying methods without freezing |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5678590A (en) * | 1979-11-05 | 1981-06-27 | Beecham Group Ltd | Enzyme derivative |
-
1988
- 1988-09-29 JP JP63245485A patent/JPH0296536A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5678590A (en) * | 1979-11-05 | 1981-06-27 | Beecham Group Ltd | Enzyme derivative |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1994015631A1 (en) * | 1993-01-11 | 1994-07-21 | The Green Cross Corporation | Dry plasminogen preparation |
| FR2719479A1 (en) * | 1994-05-04 | 1995-11-10 | Sanofi Elf | Lyophilized stable formulation comprising a protein: assay kit. |
| EP0682944A1 (en) * | 1994-05-04 | 1995-11-22 | Sanofi | Stable lyophilized formulation containing a protein and dosage kit |
| US5763409A (en) * | 1994-05-04 | 1998-06-09 | Sanofi | Stable freeze-dried formulation comprising a protein assay kit |
| CN1088583C (en) * | 1994-05-04 | 2002-08-07 | 赛诺菲-圣德拉堡股份有限公司 | Stable freeze-dried formulation comprising a protein assay kit |
| US7172999B2 (en) | 1995-10-25 | 2007-02-06 | Roche Diagnostics Gmbh | Method and preparations for stabilizing biological materials by drying methods without freezing |
| FR2740686A1 (en) * | 1995-11-03 | 1997-05-09 | Sanofi Sa | STABLE LYOPHILIZED PHARMACEUTICAL FORMULATION |
| WO1997017064A1 (en) * | 1995-11-03 | 1997-05-15 | Sanofi | Stable freeze-dried pharmaceutical formulation |
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