JPH04270227A - Composition-containing human tissue plasminogen activating factor - Google Patents
Composition-containing human tissue plasminogen activating factorInfo
- Publication number
- JPH04270227A JPH04270227A JP91115691A JP11569191A JPH04270227A JP H04270227 A JPH04270227 A JP H04270227A JP 91115691 A JP91115691 A JP 91115691A JP 11569191 A JP11569191 A JP 11569191A JP H04270227 A JPH04270227 A JP H04270227A
- Authority
- JP
- Japan
- Prior art keywords
- tpa
- modified
- human tissue
- tissue plasminogen
- 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
- 239000000203 mixture Substances 0.000 title claims abstract description 21
- 102000013566 Plasminogen Human genes 0.000 title claims 2
- 108010051456 Plasminogen Proteins 0.000 title claims 2
- 230000003213 activating effect Effects 0.000 title 1
- 150000001412 amines Chemical class 0.000 claims abstract description 16
- 102000003978 Tissue Plasminogen Activator Human genes 0.000 claims abstract description 12
- 108090000373 Tissue Plasminogen Activator Proteins 0.000 claims abstract description 12
- 229960000187 tissue plasminogen activator Drugs 0.000 claims abstract description 12
- 125000000217 alkyl group Chemical group 0.000 claims abstract description 4
- 150000003839 salts Chemical class 0.000 claims description 21
- NTNZTEQNFHNYBC-UHFFFAOYSA-N ethyl 2-aminoacetate Chemical compound CCOC(=O)CN NTNZTEQNFHNYBC-UHFFFAOYSA-N 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- UZCXPYDBYUEZCV-UHFFFAOYSA-N methyl 3-aminopropanoate Chemical group COC(=O)CCN UZCXPYDBYUEZCV-UHFFFAOYSA-N 0.000 claims 1
- KQSSATDQUYCRGS-UHFFFAOYSA-N methyl glycinate Chemical group COC(=O)CN KQSSATDQUYCRGS-UHFFFAOYSA-N 0.000 claims 1
- XNOLRFVMWOSFSK-UHFFFAOYSA-N propyl 2-aminoacetate Chemical compound CCCOC(=O)CN XNOLRFVMWOSFSK-UHFFFAOYSA-N 0.000 claims 1
- JYSNRZNGUVBJQZ-UHFFFAOYSA-N propyl 3-aminopropanoate Chemical compound CCCOC(=O)CCN JYSNRZNGUVBJQZ-UHFFFAOYSA-N 0.000 claims 1
- 241000894007 species Species 0.000 claims 1
- GSQBIOQCECCMOQ-UHFFFAOYSA-N β-alanine ethyl ester Chemical compound CCOC(=O)CCN GSQBIOQCECCMOQ-UHFFFAOYSA-N 0.000 claims 1
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 abstract description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 12
- 230000007935 neutral effect Effects 0.000 abstract description 9
- 239000004471 Glycine Substances 0.000 abstract description 7
- 239000002904 solvent Substances 0.000 abstract description 5
- 238000002360 preparation method Methods 0.000 abstract description 4
- 150000001875 compounds Chemical class 0.000 abstract description 2
- QOSMNYMQXIVWKY-UHFFFAOYSA-N Propyl levulinate Chemical compound CCCOC(=O)CCC(C)=O QOSMNYMQXIVWKY-UHFFFAOYSA-N 0.000 abstract 1
- 125000004494 ethyl ester group Chemical group 0.000 abstract 1
- 235000001014 amino acid Nutrition 0.000 description 8
- 150000001413 amino acids Chemical class 0.000 description 8
- 239000000243 solution Substances 0.000 description 7
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical class OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 6
- 239000000654 additive Substances 0.000 description 5
- 230000000694 effects Effects 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 239000008363 phosphate buffer Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- OFOBLEOULBTSOW-UHFFFAOYSA-N Malonic acid Chemical compound OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 3
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- -1 mannitol and glucose Chemical class 0.000 description 3
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 3
- 229920000053 polysorbate 80 Polymers 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- TXTWXQXDMWILOF-UHFFFAOYSA-N (2-ethoxy-2-oxoethyl)azanium;chloride Chemical compound [Cl-].CCOC(=O)C[NH3+] TXTWXQXDMWILOF-UHFFFAOYSA-N 0.000 description 2
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical class Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 2
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 235000004279 alanine Nutrition 0.000 description 2
- UCMIRNVEIXFBKS-UHFFFAOYSA-N beta-alanine Chemical compound NCCC(O)=O UCMIRNVEIXFBKS-UHFFFAOYSA-N 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 125000001147 pentyl group Chemical group C(CCCC)* 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 229920000136 polysorbate Polymers 0.000 description 2
- 230000007928 solubilization Effects 0.000 description 2
- 238000005063 solubilization Methods 0.000 description 2
- 235000000346 sugar Nutrition 0.000 description 2
- 150000008163 sugars Chemical class 0.000 description 2
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 2
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 1
- 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
- BMYNFMYTOJXKLE-UHFFFAOYSA-N 3-azaniumyl-2-hydroxypropanoate Chemical class NCC(O)C(O)=O BMYNFMYTOJXKLE-UHFFFAOYSA-N 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 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 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-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
- 229920002153 Hydroxypropyl cellulose Polymers 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
- 229930195725 Mannitol Natural products 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
- 235000011054 acetic acid Nutrition 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 125000005907 alkyl ester group Chemical group 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 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
- 229940000635 beta-alanine Drugs 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
- 230000004071 biological effect Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000006172 buffering agent Substances 0.000 description 1
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 1
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000015165 citric acid Nutrition 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 235000019800 disodium phosphate Nutrition 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000001530 fumaric acid Substances 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000011521 glass Substances 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
- 125000004051 hexyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 239000001863 hydroxypropyl cellulose Substances 0.000 description 1
- 235000010977 hydroxypropyl cellulose Nutrition 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 description 1
- 125000004491 isohexyl group Chemical group C(CCC(C)C)* 0.000 description 1
- 125000001972 isopentyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000002075 main ingredient Substances 0.000 description 1
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 1
- 239000011976 maleic acid Substances 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 229940098779 methanesulfonic acid Drugs 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 239000011707 mineral Chemical class 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 235000006408 oxalic acid Nutrition 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- OXNIZHLAWKMVMX-UHFFFAOYSA-N picric acid Chemical compound OC1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O OXNIZHLAWKMVMX-UHFFFAOYSA-N 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 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
- 239000008213 purified water Substances 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 125000002914 sec-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
- 235000002906 tartaric acid Nutrition 0.000 description 1
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 125000001973 tert-pentyl group Chemical group [H]C([H])([H])C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 239000008215 water for injection Substances 0.000 description 1
Landscapes
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
Description
【０００１】0001
【産業上の利用分野】本発明は、ヒト組織プラスミノー
ゲン活性化因子（以下ｔＰＡ と称する）や該物質を
一部改変した物質（以下改変型ｔＰＡと称する）を含有
する組成物に関する。更に詳しくは、本発明はｔＰＡ又
は改変型ｔＰＡと、特定のアミンやその塩とを含有して
なる組成物に関する。FIELD OF THE INVENTION The present invention relates to a composition containing human tissue plasminogen activator (hereinafter referred to as tPA) or a partially modified substance (hereinafter referred to as modified tPA). More specifically, the present invention relates to a composition containing tPA or modified tPA and a specific amine or a salt thereof.
【０００２】0002
【従来の技術】ｔＰＡ又は改変型ｔＰＡは水に対して難
溶であり、水に溶解して投与する剤形の製剤化を困難に
している。従来、ｔＰＡ又は改変型ｔＰＡを含有する組
成物の溶解度を高める方としては、溶液のｐＨを３〜５
とする方法及びアミノ酸又はその塩を可溶化剤として用
いる方法が知られている（特開昭６２−２６２３３号、
同６２−３６３３２号及び同６４−４２４４２号等）。BACKGROUND OF THE INVENTION tPA or modified tPA is sparingly soluble in water, making it difficult to formulate a dosage form that can be administered by dissolving it in water. Conventionally, the solubility of compositions containing tPA or modified tPA has been increased by adjusting the pH of the solution to 3 to 5.
A method using an amino acid or a salt thereof as a solubilizing agent is known (Japanese Patent Application Laid-Open No. 62-26233,
No. 62-36332 and No. 64-42442, etc.).
【０００３】溶液のｐＨを低くする方法は、その溶液状
態でｔＰＡ等が速やかに分解され失活してしまう問題点
がある。また、アミノ酸やその塩を用いる方法は、アル
ギニンなど塩基性アミノ酸やその塩を用いるときは良好
な可溶化効果を示すが、グリシン、アラニンやそれらの
塩など中性アミノ酸やその塩を用いるときは、実際上溶
解度の劣ることを確認した。The method of lowering the pH of the solution has the problem that tPA and the like are rapidly decomposed and deactivated in the solution state. In addition, methods using amino acids and their salts show good solubilization effects when basic amino acids such as arginine and their salts are used, but when using neutral amino acids and their salts such as glycine, alanine, and their salts, It was confirmed that the solubility was actually poor.
【０００４】0004
【発明が解決しようとする課題】本発明の目的は、ｔＰ
Ａ 又は改変型ｔＰＡを安定に保つ中性ｐＨ領域で、
ｔＰＡや改変型ｔＰＡの水に対する溶解性が高められた
、これらｔＰＡの新規な組成物を提供することにある。[Problems to be Solved by the Invention] An object of the present invention is to
A or in the neutral pH range that keeps modified tPA stable,
The object of the present invention is to provide a novel composition of tPA or modified tPA in which the solubility of these tPAs in water is increased.
【０００５】[0005]
【課題を解決するための手段】本発明者らは、ｔＰＡや
改変型ｔＰＡを安定に保つ中性領域でこれらの物質の溶
解度を高める可溶化剤を加えての製剤化について種々の
検討を行なった。その結果、グリシンやβ−アラニンな
ど特定のアミノ酸のアルキルエステルやその塩など特定
アミン類やその塩を可溶化剤とするときは、それらアミ
ノ酸やその塩よりも、ｔＰＡや改変型ｔＰＡの溶解性を
改善し、臨床的使用を可能にする溶解性を示し製剤化の
目的を達成することを知見し、本発明を完成させるに至
った。本発明の特定アミン類は下式（Ｉ）で示される。
Ｈ２Ｎ（ＣＨ２）ｎＣＯＯＲ （Ｉ
）（式中の記号は以下の意味を有する。
ｎ：１ないし３の整数
Ｒ：低級アルキル基）[Means for Solving the Problems] The present inventors have conducted various studies on the formulation of tPA and modified tPA by adding a solubilizer that increases the solubility of these substances in the neutral region to keep them stable. Ta. As a result, when specific amines and their salts, such as alkyl esters of specific amino acids such as glycine and β-alanine, and their salts are used as solubilizers, the solubility of tPA and modified tPA is lower than that of these amino acids and their salts. The present inventors have discovered that the present invention can be achieved by improving the solubility and achieving the purpose of formulation by exhibiting solubility that enables clinical use. The specific amines of the present invention are represented by the following formula (I). H2N(CH2)nCOOR(I
) (Symbols in the formula have the following meanings. n: integer from 1 to 3 R: lower alkyl group)
【０００６】すなわち、本発明は、ｔＰＡ又は改変型ｔ
ＰＡと前記一般式（Ｉ）で示されるアミン及びその塩か
らなる群より選択された一種又は二種以上とを含有して
なるｔＰＡ又は改変型ｔＰＡ含有組成物である。以下、
本発明化合物につき詳述する。本発明の上記一般式（Ｉ
）において、Ｒが示す低級アルキル基としては、具体的
にはメチル基、エチル基、プロピル基、イソプロピル基
、ブチル基、イソブチル基、Ｓｅｃ−ブチル基、ｔｅｒ
ｔ−ブチル基、ペンチル基、イソペンチル基、ｎｅｏ−
ペンチル基、ｔｅｒｔ−ペンチル基、ヘキシル基、イソ
ヘキシル基等が挙げられる。本発明においてｔＰＡとは
、天然型ｔＰＡの他、遺伝子工学的手法により得られた
ｔＰＡをも含む。また、本発明において改変型ｔＰＡは
、ｔＰＡの難点を改善しｔＰＡの有する生物学的活性な
どを高めたものであって、水に対する溶解度が低く、低
いｐＨでは分解され易く、かつ本発明の組成によって中
性ｐＨ領域における水溶性が高められるものであれば、
いずれの改変型ｔＰＡであってもよい。[0006] That is, the present invention provides tPA or modified t
This is a tPA or modified tPA-containing composition containing PA and one or more selected from the group consisting of amines represented by the general formula (I) and salts thereof. below,
The compound of the present invention will be explained in detail. The above general formula (I
), the lower alkyl group represented by R is specifically a methyl group, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, Sec-butyl group, ter
t-butyl group, pentyl group, isopentyl group, neo-
Examples include pentyl group, tert-pentyl group, hexyl group, isohexyl group, and the like. In the present invention, tPA includes not only natural tPA but also tPA obtained by genetic engineering techniques. In addition, the modified tPA of the present invention improves the disadvantages of tPA and increases the biological activity of tPA, has low solubility in water, is easily decomposed at low pH, and has a composition of the present invention. If the water solubility in the neutral pH region is increased by
Any modified tPA may be used.
【０００７】このような改変型ｔＰＡとしては、本出願
人の出願に係る特願昭６３−２７２０２０号、特願平１
−３１９４３８号などに示された改変型ｔＰＡなどが挙
げられるが、これらに限定されるものではない。また、
本発明で用いられる特定アミンは、一般式（Ｉ）で示さ
れるアミンであれば、いずれも中性ｐＨにおけるｔＰＡ
や改変型ｔＰＡの水に対する溶解性を高めることができ
るが、中でもグリシンエチルエステルが好ましい。この
特定アミンの塩としては、塩酸、臭化水素酸、リン酸、
硫酸、硝酸などの鉱酸、ギ酸、酢酸、プロピオン酸、シ
ュウ酸、マロン酸、コハク酸、フマール酸、マレイン酸
、乳酸、リンゴ酸、クエン酸、酒石酸、炭酸、ピクリン
酸、メタンスルホン酸などの有機酸との酸付加塩やグル
タミン酸、アスパラギン酸などの酸性アミノ酸との塩や
アンモニウム塩が挙げられる。[0007] Such modified tPA is disclosed in Japanese Patent Application No. 1983-272020 and Japanese Patent Application No. 1999 filed by the present applicant.
Examples include, but are not limited to, modified tPA shown in No.-319438. Also,
The specific amine used in the present invention is tPA at neutral pH, as long as it is an amine represented by general formula (I).
Among them, glycine ethyl ester is preferred. Salts of this specific amine include hydrochloric acid, hydrobromic acid, phosphoric acid,
Mineral acids such as sulfuric acid, nitric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, citric acid, tartaric acid, carbonic acid, picric acid, methanesulfonic acid, etc. Examples include acid addition salts with organic acids, salts with acidic amino acids such as glutamic acid and aspartic acid, and ammonium salts.
【０００８】本発明の組成物は、ｔＰＡ又は改変型ｔＰ
Ａと、前記特定アミン類及びその塩からなる群より選択
された一種又は二種以上とを配合することにより調整さ
れ、通常滅菌下これを水（精製水）に溶解してｔＰＡ又
は改変型ｔＰＡの液剤あるいはこれを更に凍結乾燥など
の乾燥手段で乾燥して用時溶解型製剤とする。ここに、
ｔＰＡ又は改変型ｔＰＡの濃度は少なくとも約０．１ｍ
ｇ／ｍｌ以上であり、一方ｔＰＡ又は改変型ｔＰＡに対
する特定アミンやその塩類の濃度は０．０１〜１．０Ｍ
、好ましくは０．１〜０．５Ｍである。本発明の組成物
には、医薬製剤化において通常使用される添加剤を、随
意配合することができ、また配合した方が好ましい。[0008] The composition of the present invention comprises tPA or modified tP.
It is prepared by blending A with one or more selected from the group consisting of the above-mentioned specific amines and their salts, and is usually dissolved in water (purified water) under sterilization to produce tPA or modified tPA. The liquid preparation or this is further dried by drying means such as freeze-drying to form a preparation that dissolves at the time of use. Here,
The concentration of tPA or modified tPA is at least about 0.1 m
g/ml or more, while the concentration of specific amines and their salts for tPA or modified tPA is 0.01-1.0M
, preferably 0.1 to 0.5M. Additives commonly used in pharmaceutical formulations can be optionally added to the composition of the present invention, and it is preferable to add them.
【０００９】このような添加剤としては、トリス塩酸緩
衝液、リン酸などの緩衝液としうる緩衝化剤、マンニト
ール、ブドウ糖などの糖類の安定化剤、塩化ナトリウム
、リン酸一水素ナトリウムなどの無機塩類の電解質等の
等張化剤、ポリオキシエチレン脂肪酸エステルなどの界
面活性化剤、ヒドロキシプロピルセルロースなどの賦形
剤などが挙げられる。前記糖類は賦形剤としても使用で
きる。これらの添加剤は適宜選択され、同種のものが二
種以上であってもよい。これら添加剤や必要により含ま
れている水の使用量は、先のｔＰＡ、改変型ｔＰＡ、特
定アミンやその塩の残りの部分である。なを、用時溶解
型の製剤とするときは、溶解液に可溶化剤を加えたり、
緩衝液とすることもできる。Such additives include buffering agents such as Tris-HCl buffer and phosphoric acid, stabilizers for sugars such as mannitol and glucose, and inorganic additives such as sodium chloride and sodium monohydrogen phosphate. Examples include tonicity agents such as salt electrolytes, surfactants such as polyoxyethylene fatty acid esters, and excipients such as hydroxypropyl cellulose. The sugars can also be used as excipients. These additives are appropriately selected, and two or more of the same type may be used. The amounts of these additives and water included if necessary are the remaining portions of the above-mentioned tPA, modified tPA, specific amine, and its salt. When making preparations that dissolve at the time of use, a solubilizer may be added to the solution, or
It can also be a buffer solution.
【００１０】0010
【実施例】次に、本発明の実施例をあげて本発明を具体
的に説明する。[Examples] Next, the present invention will be specifically explained with reference to Examples.
【００１１】実施例１
改変型ｔＰＡを１ｍｇ／ｍｌの濃度で含む、０．２Ｍ
グリシンエチルエステル塩酸塩及び０．０１％ツイー
ン８０（商品名 花王アトラス社製、以下Ｔｗｅｅｎ
８０と略記する）含有０．１Ｍリン酸２ナトリウム緩衝
液（ｐＨ７．２）を調製し、その液をガラス製バイアル
に２ｍｌずつ分注した後、共和真空技術社製の凍結乾燥
機を用いて用時溶解型の凍結乾燥品を製造した。得られ
た凍結乾燥品を注射用水で再溶解したところ、透明な液
となった。また、Ｃｌｏｔ Ｌｙｓｉｓ法による力価
試験の結果、主剤の力価も保たれていた。Example 1 0.2M containing modified tPA at a concentration of 1mg/ml
Glycine ethyl ester hydrochloride and 0.01% Tween 80 (product name: Kao Atlas Co., Ltd., hereinafter referred to as Tween)
Prepare a 0.1 M disodium phosphate buffer (pH 7.2) containing (abbreviated as 80), dispense the solution into glass vials in 2 ml portions, and then use a freeze dryer manufactured by Kyowa Vacuum Technology Co., Ltd. A freeze-dried product was produced that can be dissolved before use. When the obtained freeze-dried product was redissolved in water for injection, it became a clear liquid. Furthermore, as a result of a potency test using the Clot Lysis method, the potency of the main ingredient was also maintained.
【００１２】0012
【発明の効果】本発明は、公知のグリシンやアラニン又
はその塩など中性アミノ酸又はその塩に比較して優れた
溶解度を示し、ｔＰＡや改変型ｔＰＡを安定に保つ中性
ｐＨ領域でこれらの水に対する溶解性を高めた組成物を
提供したものである。従って、本発明組成物はｔＰＡや
改変型ｔＰＡの臨床上使用しうる製剤として有用である
。また、本発明組成物によるｔＰＡや改変型ｔＰＡの溶
解性の向上は以下の試験法によって確認されたものであ
る。Effects of the Invention The present invention exhibits superior solubility compared to known neutral amino acids such as glycine, alanine, or their salts, and their salts in a neutral pH range that keeps tPA and modified tPA stable. The present invention provides a composition with improved solubility in water. Therefore, the composition of the present invention is useful as a clinically usable formulation of tPA or modified tPA. Furthermore, the improvement in solubility of tPA and modified tPA by the composition of the present invention was confirmed by the following test method.
【００１３】実験例１
天然型ｔＰＡの凍結乾燥品を、０．０１％のＴｗｅｅｎ
８０を含むｐＨ７．０の５０ｍＭリン酸緩衝液を加えて
よく振り混ぜた。これを遠心分離し、上清を採取してた
ん白質濃度を測定した。こうして求められた天然型ｔＰ
Ａの溶解度は、０．０６ｍｇ／ｍｌであった。ｐＨを７
．０に保ったままリン酸の濃度を１００ｍＭ、５００ｍ
Ｍとした場合にもそれぞれの溶液中の天然型ｔＰＡの溶
解度は０．１３ｍｇ／ｍｌ、０．２５ｍｇ／ｍｌと低い
ものであった。また改変型ｔＰＡ（国際公開第８９／０
３８７４ 号の改良型ｔＰＡ〔ＩＶ〕を精製したもの
）のｐＨ７．０の５０ｍＭリン酸塩緩衝液中の溶解度は
０．２４ｍｇ／ｍｌであった。Experimental Example 1 Freeze-dried natural tPA was mixed with 0.01% Tween.
A 50 mM phosphate buffer containing pH 7.0 was added and the mixture was shaken thoroughly. This was centrifuged, the supernatant was collected, and the protein concentration was measured. Natural tP thus determined
The solubility of A was 0.06 mg/ml. pH to 7
．． The concentration of phosphoric acid was 100mM and 500mM while keeping it at 0.
Even when M was used, the solubility of natural tPA in each solution was as low as 0.13 mg/ml and 0.25 mg/ml. In addition, modified tPA (International Publication No. 89/0
The solubility of the purified improved tPA [IV] of No. 3874) in 50 mM phosphate buffer at pH 7.0 was 0.24 mg/ml.
【００１４】次に、グリシンエチルエステル塩酸塩をそ
れぞれ０．１Ｍ、０．２Ｍ及び０．５Ｍ含む、０．０１
％Ｔｗｅｅｎ８０含有０．０２Ｍリン酸緩衝液（ｐＨ７
．２）を用いて、ｔＰＡ及び改変型ｔＰＡについて同様
の実験を行った。その結果を図１に示す。このときグリ
シンエチルエステルを０．５Ｍ含む液は透明であったた
め、限外濾過によりさらに濃縮を行ったところ、ｔＰＡ
及び改変型ｔＰＡのいずれも２０ｍｇ／ｍｌの濃度に達
しても析出を起こさなかった。以上の結果より、本発明
に記すアミン類及びその塩がｔＰＡ及び改変型ｔＰＡの
溶解度を高める効果を有することは明らかである。一方
、グリシンを用いて同様の実験を行ったところ、グリシ
ンを０．５Ｍ含む０．０１％Ｔｗｅｅｎ８０含有０．０
２Ｍリン酸緩衝液（ｐＨ７．２）中でのｔＰＡ及び改変
型ｔＰＡの溶解度はいずれも１ｍｇ／ｍｌ以下であった
。これらの結果より、グリシンをエステル化することに
より可溶化効果が飛躍的に高められることが判明した。Next, 0.01M, 0.2M and 0.5M of glycine ethyl ester hydrochloride, respectively.
0.02M phosphate buffer containing % Tween 80 (pH 7)
．． Similar experiments were conducted using tPA and modified tPA using 2). The results are shown in Figure 1. At this time, the solution containing 0.5M glycine ethyl ester was transparent, so when it was further concentrated by ultrafiltration, tPA
Neither tPA nor modified tPA caused precipitation even when the concentration reached 20 mg/ml. From the above results, it is clear that the amines and salts thereof according to the present invention have the effect of increasing the solubility of tPA and modified tPA. On the other hand, when similar experiments were conducted using glycine, 0.01% Tween 80 containing 0.5M glycine was used.
The solubility of both tPA and modified tPA in 2M phosphate buffer (pH 7.2) was 1 mg/ml or less. These results revealed that the solubilization effect can be dramatically enhanced by esterifying glycine.
【図１】図１はｔＰＡ及び改変型ｔＰＡに対するグリシ
ンエチルエステルの可溶化効果の実験結果を示す。FIG. 1 shows experimental results of the solubilizing effect of glycine ethyl ester on tPA and modified tPA.
Claims (3)
又は該物質を一部改変した物質と、下記一般式（Ｉ）Ｈ
２Ｎ（ＣＨ２）ｎＣＯＯＲ （Ｉ）（式中の
記号は以下の意味を有する。 ｎ：１ないし３の整数 Ｒ：低級アルキル基） で示されるアミン及びその塩からなる群より選択された
一種又は二種以上とを含有してなるヒト組織プラスミノ
ーゲン活性化因子又は該物質を一部改変した物質の含有
組成物。Claim 1: Human tissue plasminogen activator or a partially modified substance thereof, and the following general formula (I)H
2N(CH2)nCOOR (I) (The symbols in the formula have the following meanings. n: An integer of 1 to 3 R: Lower alkyl group) One or two selected from the group consisting of amines and salts thereof represented by A composition containing a human tissue plasminogen activator or a partially modified substance comprising at least one species of human tissue plasminogen.
リシンエチルエステル又はグリシンプロピルエステルで
ある請求項１記載の組成物。2. The composition according to claim 1, wherein the amine is glycine methyl ester, glycine ethyl ester or glycine propyl ester.
、β−アラニンエチルエステル又はβ−アラニンプロピ
ルエステルである請求項１記載の組成物。3. The composition according to claim 1, wherein the amine is β-alanine methyl ester, β-alanine ethyl ester or β-alanine propyl ester.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP91115691A JPH04270227A (en) | 1991-02-26 | 1991-02-26 | Composition-containing human tissue plasminogen activating factor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP91115691A JPH04270227A (en) | 1991-02-26 | 1991-02-26 | Composition-containing human tissue plasminogen activating factor |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH04270227A true JPH04270227A (en) | 1992-09-25 |
Family
ID=14668860
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP91115691A Pending JPH04270227A (en) | 1991-02-26 | 1991-02-26 | Composition-containing human tissue plasminogen activating factor |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH04270227A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2813250A1 (en) * | 2012-03-27 | 2014-12-17 | Terumo Kabushiki Kaisha | Coating composition and medical device |
-
1991
- 1991-02-26 JP JP91115691A patent/JPH04270227A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2813250A1 (en) * | 2012-03-27 | 2014-12-17 | Terumo Kabushiki Kaisha | Coating composition and medical device |
| EP2813250A4 (en) * | 2012-03-27 | 2015-08-12 | Terumo Corp | Coating composition and medical device |
| AU2013238160B2 (en) * | 2012-03-27 | 2016-03-31 | Terumo Kabushiki Kaisha | Coating composition and medical device |
| US9603974B2 (en) | 2012-03-27 | 2017-03-28 | Terumo Kabushiki Kaisha | Coating composition and medical device |
| EP3219335A1 (en) * | 2012-03-27 | 2017-09-20 | Terumo Kabushiki Kaisha | Coating composition and medical device |
| US10045960B2 (en) | 2012-03-27 | 2018-08-14 | Terumo Kabushiki Kaisha | Coating composition and medical device |
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