WO1994015631A1 - Dry plasminogen preparation - Google Patents
Dry plasminogen preparation Download PDFInfo
- Publication number
- WO1994015631A1 WO1994015631A1 PCT/JP1994/000019 JP9400019W WO9415631A1 WO 1994015631 A1 WO1994015631 A1 WO 1994015631A1 JP 9400019 W JP9400019 W JP 9400019W WO 9415631 A1 WO9415631 A1 WO 9415631A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- plasminogen
- preparation
- dry
- sucrose
- albumin
- Prior art date
Links
- 102000013566 Plasminogen Human genes 0.000 title claims abstract description 65
- 108010051456 Plasminogen Proteins 0.000 title claims abstract description 65
- 238000002360 preparation method Methods 0.000 title claims abstract description 38
- 150000001413 amino acids Chemical class 0.000 claims abstract description 16
- 108010088751 Albumins Proteins 0.000 claims abstract description 13
- 102000009027 Albumins Human genes 0.000 claims abstract description 13
- 229930006000 Sucrose Natural products 0.000 claims abstract description 13
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims abstract description 13
- 239000005720 sucrose Substances 0.000 claims abstract description 13
- 239000002736 nonionic surfactant Substances 0.000 claims abstract description 8
- 238000001035 drying Methods 0.000 claims description 4
- 230000000694 effects Effects 0.000 abstract description 15
- 238000004090 dissolution Methods 0.000 abstract description 7
- 239000000126 substance Substances 0.000 abstract description 6
- 208000007536 Thrombosis Diseases 0.000 abstract description 3
- 239000003125 aqueous solvent Substances 0.000 abstract description 2
- 230000007774 longterm Effects 0.000 abstract description 2
- 238000000034 method Methods 0.000 description 19
- 239000000243 solution Substances 0.000 description 15
- 235000001014 amino acid Nutrition 0.000 description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 10
- 239000000203 mixture Substances 0.000 description 9
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 8
- 239000004472 Lysine Substances 0.000 description 8
- 239000002245 particle Substances 0.000 description 8
- 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
- 239000002904 solvent Substances 0.000 description 6
- 241000700605 Viruses Species 0.000 description 5
- 150000001720 carbohydrates Chemical class 0.000 description 5
- 238000009472 formulation Methods 0.000 description 5
- 238000004108 freeze drying Methods 0.000 description 5
- 238000010438 heat treatment Methods 0.000 description 5
- 229920000642 polymer Polymers 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- 239000000654 additive Substances 0.000 description 4
- 230000000996 additive effect Effects 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
- 239000003381 stabilizer Substances 0.000 description 4
- 238000011282 treatment Methods 0.000 description 4
- 239000004471 Glycine Substances 0.000 description 3
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 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
- -1 fatty acid ester Chemical class 0.000 description 3
- 238000005194 fractionation Methods 0.000 description 3
- 230000002779 inactivation Effects 0.000 description 3
- 229910052708 sodium Inorganic materials 0.000 description 3
- 239000011734 sodium Substances 0.000 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 description 2
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 229930195725 Mannitol Natural products 0.000 description 2
- 229920002684 Sepharose Polymers 0.000 description 2
- 235000021355 Stearic acid Nutrition 0.000 description 2
- 241000209149 Zea Species 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 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
- 235000005822 corn Nutrition 0.000 description 2
- 235000014113 dietary fatty acids Nutrition 0.000 description 2
- 150000002016 disaccharides Chemical class 0.000 description 2
- POULHZVOKOAJMA-UHFFFAOYSA-N dodecanoic acid Chemical compound CCCCCCCCCCCC(O)=O POULHZVOKOAJMA-UHFFFAOYSA-N 0.000 description 2
- 229930195729 fatty acid Natural products 0.000 description 2
- 239000000194 fatty acid Substances 0.000 description 2
- 230000020764 fibrinolysis Effects 0.000 description 2
- 235000013922 glutamic acid Nutrition 0.000 description 2
- 239000004220 glutamic acid Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 125000003588 lysine group Chemical group [H]N([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 2
- 239000000594 mannitol Substances 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 2
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 2
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 description 2
- 229940012957 plasmin Drugs 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000008117 stearic acid Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-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
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 1
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 108010039627 Aprotinin Proteins 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 206010003445 Ascites Diseases 0.000 description 1
- 239000005635 Caprylic acid (CAS 124-07-2) Substances 0.000 description 1
- 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 description 1
- 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 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
- 241000282412 Homo Species 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
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products 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 1
- 239000005639 Lauric acid Substances 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 239000005642 Oleic acid Substances 0.000 description 1
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- RVGRUAULSDPKGF-UHFFFAOYSA-N Poloxamer Chemical compound C1CO1.CC1CO1 RVGRUAULSDPKGF-UHFFFAOYSA-N 0.000 description 1
- 229920001214 Polysorbate 60 Polymers 0.000 description 1
- 239000002253 acid Substances 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
- 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
- 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
- 239000007853 buffer solution Substances 0.000 description 1
- 238000011284 combination treatment Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 239000003527 fibrinolytic agent Substances 0.000 description 1
- 230000003480 fibrinolytic effect 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
- 125000000291 glutamic acid group Chemical group N[C@@H](CCC(O)=O)C(=O)* 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 238000011534 incubation Methods 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
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 230000002934 lysing effect Effects 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 125000001360 methionine group Chemical group N[C@@H](CCSC)C(=O)* 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 229960002446 octanoic acid Drugs 0.000 description 1
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 239000008194 pharmaceutical composition Substances 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
- 230000003169 placental effect Effects 0.000 description 1
- 210000005059 placental tissue Anatomy 0.000 description 1
- 229960000502 poloxamer Drugs 0.000 description 1
- 229920001983 poloxamer Polymers 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920002503 polyoxyethylene-polyoxypropylene Polymers 0.000 description 1
- 229920000053 polysorbate 80 Polymers 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
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 150000005846 sugar alcohols Chemical class 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 230000001732 thrombotic effect Effects 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/43—Enzymes; Proenzymes; Derivatives thereof
- A61K38/46—Hydrolases (3)
- A61K38/48—Hydrolases (3) acting on peptide bonds (3.4)
- A61K38/482—Serine endopeptidases (3.4.21)
- A61K38/484—Plasmin (3.4.21.7)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
- A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
Definitions
- the present invention relates to an improvement in a plasminogen preparation. More specifically, the present invention relates to a plasminogen dry preparation in which the solubility of the plasminogen dry preparation in water is improved.
- Plasminogen is activated by perokinase and the like to become plasmin, which dissolves fibrin and causes fibrinolysis. Plasminogen is used not only for the study of fibrinolysis phenomena, but also in clinical applications as a fibrinolytic therapeutic agent (thrombotic agent) in recent years.
- plasminogen itself is a precursor enzyme, and is stable unless plasmin or the like is not contained as a contaminant in the preparation. Plasminogen has poor solubility and dissolution in water by making it into a dry preparation.c.Applicant must first add a dry preparation of plasminogen to A plasminogen preparation to which sugar or amino acid is added as a solubilizing agent for suppressing the generation has been proposed (Japanese Patent Application Laid-Open No. Sho 62-153324).
- An object of the present invention is to provide a dry plasminogen preparation which has good solubility and dissolution in water of dried plasminogen even under severe conditions. Disclosure of the invention
- the first invention of the present invention is a dried plasminogen preparation comprising plasminogen and a nonionic surfactant.
- the second invention of the present invention is a dried plasminogen preparation comprising plasminogen, sucrose, amino acid and albumin.
- the plasminogen dry preparation in the present invention refers to a preparation in a substantially dried state, and may contain a certain amount of water, for example, to the extent that it can be made into a powder.
- the dried plasminogen preparation of the present invention contains dried plasminogen as a main component.
- the dried plasminogen include a powdered form, particularly, lyophilized plasminogen.
- Such a dried plasminogen is produced by drying a plasminogen-containing liquid by a known means, particularly by freeze-drying.
- the plasminogen-containing solution is not particularly limited.
- serum III, plasma, ascites, placental extract, placental tissue extract, and corn fraction III of the low-temperature alcohol fractionation method of animals including humans As described above, a plasminogen purified from a plasminogen-containing fraction by a widely known method, a solution containing plasminogen obtained by genetic engineering, and the like can be mentioned.
- Examples of the plasminogen of the present invention include a methionyl type (the N-terminal of which is methionine), a glutamyl type (the N-terminal of which is glutamic acid), and a lysyl type (the N-terminal of which is lysine). Further, a mixture thereof may be used.
- Preferred is lysyl monoplasminogen.
- Representative methods for purifying plasminogen include, for example, the method described in Japanese Patent Application Laid-Open No. 55-159392 and the purification method using lysine-sepharose (Science, 170, 1095, 1970). It can be exemplified as a simple method.
- the plasminogen preparation has been treated for virus inactivation.
- virus inactivation treatment include a method of heating a liquid composition of plasminogen at 50 to 100 ° C for 5 to 30 hours, and a method of heating a dry composition at 50 to 100 ° C for 10 to 1 hour. 50 hours heating method, surfactant contact method, other chemicals such as trialkyl phosphate contact method and treatment Combinations are included.
- the solubilizer used in the first invention is a nonionic surfactant.
- the nonionic surfactant include polyoxyethylene sorbitan fatty acid ester (twin type), polyoxyethylene-polyoxypropylene copolymer (pull nick type) and the like.
- the former fatty acids include stearic acid, panolemitic acid, oleic acid, lauric acid, stearic acid and the like.
- the latter has a molecular weight of about 2000 to 2000.
- a saccharide and an amino acid as a solubilizing agent in addition to the nonionic surfactant.
- saccharide examples include monosaccharides (such as glucose and galactose), disaccharides (such as sucrose and lactose), and sugar alcohols (such as mannitol and sorbitol).
- monosaccharides such as glucose and galactose
- disaccharides such as sucrose and lactose
- sugar alcohols such as mannitol and sorbitol.
- Preferred saccharides are disaccharides, and sucrose is particularly preferable. It is.
- amino acids examples include neutral amino acids (such as glycine and alanine), acidic amino acids (such as glutamic acid and aspartic acid), and basic amino acids (such as arginine and lysine).
- neutral amino acids such as glycine and alanine
- acidic amino acids such as glutamic acid and aspartic acid
- basic amino acids such as arginine and lysine.
- lysine is used.
- the dried plasminogen preparation of the present invention is usually prepared by adding a solubilizer to a plasminogen solution and subjecting it to a known drying treatment.
- the compounding amount of the solubilizer is an amount sufficient to suppress the generation of a filamentous substance when dissolving the plasminogen dry preparation of the present invention in water or the like, and specifically, It is as follows for each component.
- the amount of the nonionic surfactant to be used is about 0.01 to 0.1% (W / V), more preferably 0.01 to 0.4, based on 100 to 500 CU / ml of plasminogen. % (W / V).
- the amount of saccharide and amino acid to be used is preferably about 0.005% (W / V) to 10% (W / V), respectively, based on 100 to 500 CUZml of plasminogen. Or about 0.1 to 5% (WZV).
- the preferred weight ratio of the saccharide [especially, sucrose] (A) to the basic amino acid [especially, lysine] (B), that is, BZA is 0.001 to 0.2, and most preferably 0%. . 0
- the lysing agents used in the second invention are sucrose, amino acids and albumin.
- the amino acid used in the second invention is preferably the same as the amino acid used in the first invention.
- Albumin is preferably human-derived albumin due to the problem of antigenicity, and there is no particular limitation as long as it is purified for medical use. Its purity is preferably such that at least 80% of albumin is analyzed by electrophoresis.
- Methods for obtaining human-derived albumin include the ethanol fractionation method (Japanese Patent Publication No. 47-2869, Japanese Patent Publication No. 35-52997), and heating in the presence of an organic acid. Methods (Japanese Patent Publication No. Sho 43-1640, Japanese Patent Publication No. Sho 51-40132) are exemplified. In particular, those obtained by heat-treating albumin (preferably heat-treating at 60 ° C for about 10 hours) and inactivating virus such as hepatitis virus are used.
- the amount of sucrose and amino acid used in the present invention is about the same as the amount used in the first invention, and the amount of albumin used is 0 to 500 to 500 CU Zml of plasminogen. It is about 0.5 to 5% (WZV), more preferably about 0.25 to 1% (W / V).
- the pH of the solution of the dried plasminogen preparation of the present invention before drying is 7.2 to 7.8.
- the plasminogen dry preparation of the present invention is administered after being dissolved in an aqueous solvent such as physiological saline or distilled water for injection before use.
- an aqueous solvent such as physiological saline or distilled water for injection before use.
- it is generally administered intraarterially or intravenously.
- 100 to 300 CU of plasminogen is administered at a time.
- an effective amount of excipients and the like may be added to the preparation of the present invention in accordance with customary techniques for pharmaceutical production.
- the adsorbed plasminogen was eluted using a solvent containing 0.2 ⁇ -aminocaproic acid and 0.9% sodium chloride ( ⁇ 7.0). .
- PH on plasminogen preparations The effect of PH on plasminogen preparations was investigated. Prepare a plasminogen solution of pH 6.4, 7.2, and 7.8 consisting of 0.4% resin, 5% sucrose, and 50 mM phosphate buffer, and incubate at 45 ° C for 3 hours. After one session, the appearance and residual activity of each solution were confirmed. The residual activity was measured by the synthetic substrate method according to the method of Kato et al. [J. Biochem., 88, 183, (1980)].
- the effect of adding sugar on the plasminogen preparation was examined. After adding 0.2% lysine and 5% sucrose or mannitol (final concentration) to the plasminogen solution and freeze-drying 61 ° C for 7 hours Dry heating ⁇ 37 ° C for 8 weeks The sample was redissolved in distilled water, and after 30 minutes, the appearance, residual activity, and polymer content were confirmed. The residual activity was measured in the same manner as in Example 2, and the polymer content was measured by the HPLC method.
- the effect of the stabilizer on the plasminogen preparation was investigated. Add a certain amount of stabilizer to a plasminogen solution consisting of 0.4% lysine, 5.0% sucrose, 50 mM sodium phosphate, and pH 7.2 to 7.8. Left for 2 weeks at ° C. After redissolving in distilled water, the appearance, residual activity ratio and polymer content were confirmed one hour later.
- Table 4 shows the results of using a stabilizer as a comparative example
- Table 5 shows the results of using a nonionic surfactant and albumin of the present invention as stabilizers.
- the plasminogen dry preparation of the present invention suppresses the production of filamentous substances when dissolved in water, and retains its activity even after dissolution. In addition, it has excellent stability during lyophilization, especially long-term stability, making it more safe and practical for use as a pharmaceutical formulation. Therefore, the preparation of the present invention can more effectively treat, for example, various thrombosis.
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Abstract
Description
明 細 書 プラスミノゲン乾燥製剤 Description Plasminogen dry preparation
技術分野 Technical field
本発明は、 プラスミノゲン製剤の改良に関する。 さらに詳しくは、 プラスミノ ゲン乾燥製剤の水への溶解性を改良したプラスミノゲン乾燥製剤に関するもので ある。 背景技術 The present invention relates to an improvement in a plasminogen preparation. More specifically, the present invention relates to a plasminogen dry preparation in which the solubility of the plasminogen dry preparation in water is improved. Background art
プラスミノゲンは、 ゥロキナーゼなどによって活性化されてプラスミ ンとなり、 これがフイブリンを溶解して線溶現象をもたらすのである。 プラスミノゲンは、 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。 Plasminogen is activated by perokinase and the like to become plasmin, which dissolves fibrin and causes fibrinolysis. Plasminogen is used not only for the study of fibrinolysis phenomena, but also in clinical applications as a fibrinolytic therapeutic agent (thrombotic agent) in recent years.
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。 し力、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 c 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溶解剤として糖またはァ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 6 2 - 1 5 3 2 2 4号公報） 。 In a plasminogen preparation, plasminogen itself is a precursor enzyme, and is stable unless plasmin or the like is not contained as a contaminant in the preparation. Plasminogen has poor solubility and dissolution in water by making it into a dry preparation.c.Applicant must first add a dry preparation of plasminogen to A plasminogen preparation to which sugar or amino acid is added as a solubilizing agent for suppressing the generation has been proposed (Japanese Patent Application Laid-Open No. Sho 62-153324).
し力、し、 プラスミノゲン含有組成物を苛酷条件下 （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。 It was found that by subjecting the plasminogen-containing composition to severe conditions (for example, treatment for virus inactivation), the solubility and dissolution of the dried preparation deteriorated.
このため、 さらにアルブミ ン、 糖およびアミノ酸を併用してなる改良乾燥製剤 を提案した。 （特開平 2— 9 6 5 3 6号公報） For this reason, we have proposed an improved dry formulation using a combination of albumin, sugar and amino acid. (Japanese Unexamined Patent Publication No. 2-96653)
本発明の目的は、 苛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ブラスミノゲン乾燥製剤を提供すること である。 発明の開示 An object of the present invention is to provide a dry plasminogen preparation which has good solubility and dissolution in water of dried plasminogen even under severe conditions. Disclosure of the invention
本発明の第 1の発明は、 プラスミノゲン及び非イオン系界面活性剤を含有して なるプラスミノゲン乾燥製剤である。 The first invention of the present invention is a dried plasminogen preparation comprising plasminogen and a nonionic surfactant.
また、 本発明の第 2の発明は、 プラスミノゲン、 ショ糖、 アミノ酸及びアルブ ミ ンを含有してなるプラスミノゲン乾燥製剤である。 The second invention of the present invention is a dried plasminogen preparation comprising plasminogen, sucrose, amino acid 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 plasminogen dry preparation in the present invention refers to a preparation in a substantially dried state, and may contain a certain amount of water, for example, to the extent that 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、 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 contains dried plasminogen as a main component. Examples of the dried plasminogen include a powdered form, particularly, lyophilized plasminogen. Such a dried plasminogen is produced by drying a plasminogen-containing liquid by a known means, particularly by freeze-drying.
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 む動物等の血清、 血漿、 腹水、 胎盤抽出液、 胎盤組織抽出液、 コーンの低温アル コール分画法の画分 I I I のようにプラスミノゲンを含有する画分から広く公知の 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 solution is not particularly limited.For example, serum III, plasma, ascites, placental extract, placental tissue extract, and corn fraction III of the low-temperature alcohol fractionation method of animals including humans As described above, a plasminogen purified from a plasminogen-containing fraction by a widely known method, a solution containing plasminogen obtained by genetic engineering, and the like can be mentioned.
本発明のプラスミノゲンは、 メチォニル型 （その N末がメチォニンである） 、 グルタミル型 （その N末がグルタミ ン酸である） またはリジル型 （その N末がリ ジンである） 等が例示される。 また、 これらの混合物であってもよい。 Examples of the plasminogen of the present invention include a methionyl type (the N-terminal of which is methionine), a glutamyl type (the N-terminal of which is glutamic acid), and a lysyl type (the N-terminal of which is lysine). Further, a mixture thereof may be used.
好ましくはリジル一プラスミノゲンである。 Preferred is lysyl monoplasminogen.
プラスミノゲンの精製法としては、 たとえば特開昭 5 5 - 1 5 3 5 9 2号公報 に記載の方法、 リジン—セファロ一スによる精製法 （Sci ence, 170， 1095， 1970 年） などを代表的な方法として例示することができる。 Representative methods for purifying plasminogen include, for example, the method described in Japanese Patent Application Laid-Open No. 55-159392 and the purification method using lysine-sepharose (Science, 170, 1095, 1970). It can be exemplified as a simple 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 5 0〜 1 0 0 °Cで 5〜 3 0時間加熱する方法、 乾燥組成物を 5 0〜 1 0 0 °C で、 1 0 ~ 1 5 0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ヱー卜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。 It is preferable that the plasminogen preparation has been treated for virus inactivation. Examples of the virus inactivation treatment include a method of heating a liquid composition of plasminogen at 50 to 100 ° C for 5 to 30 hours, and a method of heating a dry composition at 50 to 100 ° C for 10 to 1 hour. 50 hours heating method, surfactant contact method, other chemicals such as trialkyl phosphate contact method and treatment Combinations are included.
以下、 本発明の第 1の発明であるプラスミノゲン乾燥製剤について説明する。 第 1の発明で使用される溶解剤は非イオン系界面活性剤である。 この非イオン 系界面活性剤としてはポリォキシエチレンソルビタン脂肪酸エステル （トウイ一 ン系） 、 ポリオキシエチレン一ポリオキシプロピレン共重合体 （プル口ニック 系） 等が例示される。 Hereinafter, the plasminogen dry preparation according to the first invention of the present invention will be described. The solubilizer used in the first invention is a nonionic surfactant. Examples of the nonionic surfactant include polyoxyethylene sorbitan fatty acid ester (twin type), polyoxyethylene-polyoxypropylene copolymer (pull nick type) and the like.
前者の脂肪酸としてはステアリ ン酸、 パノレミチン酸、 ォレイン酸、 ラウリ ン酸、 ステアリン酸等が挙げられる。 また、 後者の分子量は 2 0 0 0〜2 0 0 0 0程度 であることが例示される。 Examples of the former fatty acids include stearic acid, panolemitic acid, oleic acid, lauric acid, stearic acid and the like. In addition, the latter has a molecular weight of about 2000 to 2000.
また、 溶解剤として、 非イオン系界面活性剤の他に糖類およびアミノ酸を加え ることがより好ましい。 It is more preferable to add a saccharide and an amino acid as a solubilizing agent in addition to the nonionic surfactant.
糖類としては単糖類 （ブドウ糖、 ガラク トースなど） 、 二糖類 （ショ糖、 乳糖 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、 好まし い糖類は二糖類であり、 特に好ましくはショ糖である。 Examples of the saccharide include monosaccharides (such as glucose and galactose), disaccharides (such as sucrose and lactose), and sugar alcohols (such as mannitol and sorbitol). Preferred saccharides are disaccharides, and sucrose is particularly preferable. It is.
アミノ酸としては、 中性ァミノ酸 （グリシン、 ァラニンなど） 、 酸性ァミノ酸 (グルタミ ン酸、 ァスパラギン酸など） 、 塩基性アミノ酸 （アルギニン、 リジン など） が例示され、 好ましくは塩基性アミノ酸であり、 特に好ましくはリジンで める。 Examples of the amino acids include neutral amino acids (such as glycine and alanine), acidic amino acids (such as glutamic acid and aspartic acid), and basic amino acids (such as arginine and lysine). Preferably, lysine is used.
本発明のプラスミノゲン乾燥製剤は、 通常溶解剤をプラスミノゲン溶液に添加 した後、 既知の乾燥処理に付すことによって調製される。 The dried plasminogen preparation of the present invention is usually prepared by adding a solubilizer to a plasminogen solution and 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等に 溶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 compounding amount of the solubilizer is an amount sufficient to suppress the generation of a filamentous substance when dissolving the plasminogen dry preparation of the present invention in water or the like, and specifically, It is as follows for each component.
非イオン系界面活性剤の使用量はプラスミノゲンの 1 0 0〜 5 0 0 CU/mlに 対して 0 0 1〜0. 1 % (W/V) 程度、 より好ましくは 0. 0 1〜 0 4 % (W/V) 程度である。 The amount of the nonionic surfactant to be used is about 0.01 to 0.1% (W / V), more preferably 0.01 to 0.4, based on 100 to 500 CU / ml of plasminogen. % (W / V).
糖類およびァミノ酸の使用量は、 それぞれプラスミノゲンの 1 0 0〜5 0 0 C UZmlに対して、 0. 0 0 5 % (W/V) 〜 1 0 % (W/V) 程度、 より好まし くは 0. 1 ~ 5 % (WZV) 程度である。 なお、 糖類 〔特に、 ショ糖〕 （A) と塩基性アミノ酸 〔特に、 リ ジン〕 （B) との好ましい重量比、 即ち BZAは 0. 0 0 1〜0. 2であり、 最適には 0. 0The amount of saccharide and amino acid to be used is preferably about 0.005% (W / V) to 10% (W / V), respectively, based on 100 to 500 CUZml of plasminogen. Or about 0.1 to 5% (WZV). The preferred weight ratio of the saccharide [especially, sucrose] (A) to the basic amino acid [especially, lysine] (B), that is, BZA is 0.001 to 0.2, and most preferably 0%. . 0
1〜0. 1である。 1 to 0.1.
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 above-mentioned range of the amount of the dissolving agent to be added, the balance between the stability of the formulation, water solubility, dissolution and formulation is the best.
次に、 本発明の第 2の発明であるプラスミノゲン乾燥製剤について説明する。 第 2の発明で使用される溶解剤はショ糖、 ァミノ酸およびアルブミ ンである。 この第 2の発明に使用されるアミノ酸は、 前記の第 1の発明で使用されるァミノ 酸と同じものが好ましい。 Next, the plasminogen dry preparation according to the second invention of the present invention will be described. The lysing agents used in the second invention are sucrose, amino acids and albumin. The amino acid used in the second invention is preferably the same as the amino acid used in the first invention.
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 8 0 %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 4 7 - 2 8 6 9号公報、 特 公昭 3 5— 5 2 9 7号公報） 、 有機酸の存在下で加熱する方法 （特公昭 4 3一 1 6 0 4号公報、 特公昭 5 1 - 4 0 1 3 2 1号公報） などが例示される。 特に、 好 ましくはアルブミ ンを加熱処理 （好ましくは、 6 0 °C、 1 0時間程度の加熱処 理） して肝炎ウィルスなどのウイルス不活性化処理を行つたものが使用される。 Albumin is preferably human-derived albumin due to the problem of antigenicity, and there is no particular limitation as long as it is purified for medical use. Its purity is preferably such that at least 80% of albumin is analyzed by electrophoresis. Methods for obtaining human-derived albumin include the ethanol fractionation method (Japanese Patent Publication No. 47-2869, Japanese Patent Publication No. 35-52997), and heating in the presence of an organic acid. Methods (Japanese Patent Publication No. Sho 43-1640, Japanese Patent Publication No. Sho 51-40132) are exemplified. In particular, those obtained by heat-treating albumin (preferably heat-treating at 60 ° C for about 10 hours) and inactivating virus such as hepatitis virus are used.
この発明に使用するショ糖およびァミノ酸の使用量は第 1の発明で使用した量 と同程度であり、 アルブミ ンの使用量は、 プラスミノゲンの 1 0 0~5 0 0 C U Zmlに対して 0. 0 5 ~ 5 % (WZV) 程度、 より好ましくは 0. 2 5〜 1 % (W/V) 程度である。 The amount of sucrose and amino acid used in the present invention is about the same as the amount used in the first invention, and the amount of albumin used is 0 to 500 to 500 CU Zml of plasminogen. It is about 0.5 to 5% (WZV), more preferably about 0.25 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 above-mentioned range of the amount of the dissolving agent to be added, the balance between the stability of the formulation, water solubility, dissolution and formulation is the best.
また、 本発明のプラスミノゲン乾燥製剤の乾燥前の溶液の P Hが 7. 2〜7. 8であることが好ましい。 Further, it is preferred that the pH of the solution of the dried plasminogen preparation of the present invention before drying is 7.2 to 7.8.
本発明のプラスミノゲン乾燥製剤は、 用時生理食塩液あるいは注射用蒸留水な どの水性溶媒に溶解されて投与される。 投与に際しては、 一般的には動脈内また は静脈内投与が行われ、 例えば血栓症の治療のためには一回当たりプラスミノゲ ンとして 1 0 0〜 3 0 0 0 CUの投与が行われる。 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。 発明を実施するための最良の形態 The plasminogen dry preparation of the present invention is administered after being dissolved in an aqueous solvent such as physiological saline or distilled water for injection before use. For administration, it is generally administered intraarterially or intravenously. For example, for treatment of thrombosis, 100 to 300 CU of plasminogen is administered at a time. In addition, an effective amount of excipients and the like may be added to the preparation of the present invention in accordance with customary techniques for pharmaceutical production. BEST MODE FOR CARRYING OUT THE INVENTION
プラスミノゲン溶液の精製 Purification of plasminogen solution
コーンの冷エタノール分画法で得られた画分 ΙΙ + ΠΙ ペースト抽出残渣を、 1 Fraction obtained by cold ethanol fractionation of corn ΙΙ + ペ ー ス ト
0単位 Znilのァプロチニンと 0. 1 M塩化ナ トリゥムを含むトリス塩酸緩衝液 ( H 8. 3) に懸濁し、 少時攪拌した後 5 %硫酸アンモニゥムを添加 '攪拌し 遠心分離により上清を分離した。 更に、 この上清に 3 0 %硫酸アンモニゥムを添 加 ·攪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 %グリシン溶液 （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一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 M塩化ナトリウムを含む 0.0 units Suspend in Tris-HCl buffer (H8.3) containing aprotinin of Znil and 0.1 M sodium chloride, agitate for a short time, then add 5% ammonium sulfate.'Stir and separate supernatant by centrifugation did. Further, 30% ammonium sulfate was added to the supernatant, stirred, and the precipitate was separated by centrifugation. This precipitate was suspended in a 0.9% glycine solution (pH 7.2) containing 0.9% sodium chloride and subjected to the method of Deutsch, DG et al. [Science, 170, 1095, (1970)]. Inject into a lysine-sepharose column as described above, adsorb plasminogen, and then add 1 M sodium chloride containing the impure protein.
9 %グリシン溶液 （pH 7. 2) で洗浄した後、 0. 2 Με—アミノカプロン酸 と 0. 9 %塩化ナトリウムとを含む溶媒 （ρΗ 7. 0) を用いて吸着したプラス ミノゲンを溶出させた。 After washing with 9% glycine solution (pH 7.2), the adsorbed plasminogen was eluted using a solvent containing 0.2Με-aminocaproic acid and 0.9% sodium chloride (ρΗ7.0). .
実施例 1 Example 1
プラスミノゲン製剤にリジン、 ショ糖を添加した場合の効果を調べた。 精製し たプラスミノゲン （非活性は 8. l x l 06 mg当たり 1. 9 5 X 1 08 c u) を 0. 2 %リジン、 5 %ショ糖、 0. 4 5 %塩化ナトリウム含有 5 0 mMリ ン酸 緩衝液 （PH 7. 2) に透析して、 溶液の外観を観察した。 また、 透析後に一 3 0°C以下で少なく とも 1週間凍結し、 融解した場合の溶液の外観を観察した。 The effect of adding lysine and sucrose to the plasminogen preparation was examined. Purified plasminogen (inactive is 8. lxl 0 6 mg per 1. 9 5 X 1 0 8 cu ) a 0.2% lysine, 5% sucrose, 0.4 5% sodium chloride containing 5 0 mM Li down The solution was dialyzed against an acid buffer (PH 7.2) and the appearance of the solution was observed. After dialysis, the solution was frozen at 130 ° C or less for at least one week, and the appearance of the solution when thawed was observed.
プラスミノゲン製剤における P Hの効果を調べた。 0. 4 %リ ジン、 5 %ショ 糖、 5 0 mMリン酸緩衝液からなる、 PH 6. 4、 7. 2及び 7. 8のプラスミ ノゲン溶液を調製し、 4 5°Cで 3時間ィンキュベ一ションした後に、 各溶液の外 観及び残存活性率を確認した。 残存活性率は、 Katoら [J. Biochem.，88， 183，（198 0)]の方法に準じ、 合成基質法により、 測定した。 The effect of PH on plasminogen preparations was investigated. Prepare a plasminogen solution of pH 6.4, 7.2, and 7.8 consisting of 0.4% resin, 5% sucrose, and 50 mM phosphate buffer, and incubate at 45 ° C for 3 hours. After one session, the appearance and residual activity of each solution were confirmed. The residual activity was measured by the synthetic substrate method according to the method of Kato et al. [J. Biochem., 88, 183, (1980)].
表 2 Table 2
(インキュベ-シヨン 直前を 1 0 0 %とする） (100% immediately before the incubation)
実施例 3 Example 3
プラスミノゲン製剤における糖の添加効果を調べた。 プラスミノゲン溶液に 0. 2 %リ ジン及び、 5 %ショ糖またはマンニトール （いずれも終濃度） を添加して 凍結乾燥 6 1 °C 7 時間乾燥加熱→ 3 7 °C 8週間保存した場合に各工程におけ る検体を蒸留水で再溶解し、 3 0分後に外観、 残存活性率および重合体含有率を 確認した。 残存活性率は実施例 2と同様の方法で、 重合体含有率は HP L C法に より測定した。 The effect of adding sugar on the plasminogen preparation was examined. After adding 0.2% lysine and 5% sucrose or mannitol (final concentration) to the plasminogen solution and freeze-drying 61 ° C for 7 hours Dry heating → 37 ° C for 8 weeks The sample was redissolved in distilled water, and after 30 minutes, the appearance, residual activity, and polymer content were confirmed. The residual activity was measured in the same manner as in Example 2, and the polymer content was measured by the HPLC method.
3 Three
(残存活性率は凍結乾燥前を 1 0 0 %とする） 実施例 4 (Remaining activity rate is 100% before freeze-drying) Example 4
プラスミノゲン製剤における安定化剤の効果を調べた。 0. 4 %リジン、 5. 0 %ショ糖、 5 0 mMリン酸ナトリウム、 p H 7. 2〜 7. 8からなるプラスミ ノゲン溶液に所定量の安定化剤を添加し、 凍結乾燥後に 6 1 °Cで 2週間置いた。 蒸留水に再溶解し、 その 1時間後に外観、 残存活性率および重合体含有率を確認 した。 表 4には比較例としての安定化剤を、 表 5には本発明としての非イオン系 界面活性剤とァルブミンを安定化剤として使用した結果を記した。 The effect of the stabilizer on the plasminogen preparation was investigated. Add a certain amount of stabilizer to a plasminogen solution consisting of 0.4% lysine, 5.0% sucrose, 50 mM sodium phosphate, and pH 7.2 to 7.8. Left for 2 weeks at ° C. After redissolving in distilled water, the appearance, residual activity ratio and polymer content were confirmed one hour later. Table 4 shows the results of using a stabilizer as a comparative example, and Table 5 shows the results of using a nonionic surfactant and albumin of the present invention as stabilizers.
表 4 Table 4
添 加 剤 外 観 重合体 残存 Additive Appearance Polymer Remaining
含有率 活性率 種類 最終濃度 色彩 混濁 不溶性物質 Content Activity Rate Type Final concentration Color Opaque Insoluble substance
無添加 微かに白色 混濁 粒子状 Z繊維状 0.51 % 107 % クェン 酸 0. 4 % 微かに白色 混濁 粒 子 状 0.70 % 72 % ナトリウム 0. 2 微かに白色 混濁 粒 子 状 0.47 124 Additive Slightly white turbid particulate Z-fiber 0.51% 107% citric acid 0.4% Slightly white turbid particle 0.70% 72% Sodium 0.2 Slightly white turbid particle 0.47 124
0. 1 微かに白色 粒 子 状 0.48 0.1 Slightly white granular 0.48
塩酸 0. 4 % 微かに白色 混濁 粒子状 Z繊維状 0.32 % 107 % ナトリウム 0. 2 微かに白色 混濁 粒子状 Z繊維状 0.37 Hydrochloric acid 0.4% Slightly white turbid Particulate Z fibrous 0.32% 107% Sodium 0.2 Slightly white turbid Particulate Z fibrous 0.37
0. 1 微かに白色 混濁 粒子状/繊維状 0.46 111 力プリル酸 133mg/ml 微かに白色 混濁 粒 子 状 1.37 % 82 % ナトリウム 66 微かに白色 混濁 粒 子 状 0.96 94 0.1 Slightly white turbid Particle / fibrous 0.46 111 Caprylic acid 133 mg / ml Slightly white turbid particle 1.37% 82% Sodium 66 Slightly white turbid particle 0.96 94
33 微かに白色 混濁 粒 子 状 0.73 90 マルト-ス 5. 0 % 黄色 澄明 粒 子 状 2.58 % 33 Slightly white turbid particles 0.73 90 Maltose 5.0% Yellow clear particles 2.58%
2. 5 黄色 澄明 粒 子 状 1.43 76 % 2.5 Yellow clear particle 1.43 76%
添 加 剤 外 観 重合体 残存 Additive Appearance Polymer Remaining
含有率 活性率 最終濃度 色彩 混濁 不溶性物質 Content Activity Final concentration Color Opaque Insoluble substance
無添加 微かに白色 混濁 粒子状ノ繊維状 0. 04 % 95 % トウィ-ン 0. 04 % 無色 澄明 観察されず 0. 25 % Additive Slightly white Cloudy Particulate fibrous 0.04% 95% Twin 0.04% Colorless and clear 0.25%
80 0 02 無色 澄明 観察されず 0. 23 93 % 80 0 02 colorless clear Not observed 0.23 93%
0. 01 無色 澄明 観察されず 0. 21 97 ポロキサマ 0. 04 % 無色 澄明 観察されず 0. 41 % 0.01 Colorless clear Not observed 0.221 97 Poloxamer 0.04% Colorless clear Not observed 0.41%
188 0. 02 無色 澄明 観察されず 0. 47 99 % 188 0.02 Colorless clear Not observed 0.47 99%
0. 01 無色 澄明 観察されず 0. 57 110 0.01 Colorless clear Not observed 0.57 110
1. 0 % 無色 澄明 観察されず 0. 09 % 91 % アルブミン 0. 5 無色 澄明 観察されず 0. 15 91 1.0% colorless clear Not observed 0.09% 91% albumin 0.5 Colorless clear Not observed 0.159
0. 25 無色 澄明 観察されず 0. 19 無添加 * 無色 澄明 観察されず 0. 04 % 1 10 % 0.25 Colorless clear No observation 0.19 No addition * Colorless clear No observation 0.04% 1 10%
(残存活性率は凍結乾燥前を 1 0 0 %とする） (Remaining activity rate is 100% before freeze-drying)
* ：蒸留水の代わりに 0 . 0 2 %トウィーン 8 0で再溶解した場合 産業上の利用可能性 *: When redissolved in 0.02% Tween 80 instead of distilled water Industrial applicability
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、 糸状様物 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 り効果的に行うことができる。 The plasminogen dry preparation of the present invention suppresses the production of filamentous substances when dissolved in water, and retains its activity even after dissolution. In addition, it has excellent stability during lyophilization, especially long-term stability, making it more safe and practical for use as a pharmaceutical formulation. Therefore, the preparation of the present invention can more effectively treat, for example, various thrombosis.
Claims
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP5/17816 | 1993-01-11 | ||
| JP5017816A JPH06211691A (en) | 1993-01-11 | 1993-01-11 | Dry form of plasminogen |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO1994015631A1 true WO1994015631A1 (en) | 1994-07-21 |
Family
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/JP1994/000019 WO1994015631A1 (en) | 1993-01-11 | 1994-01-10 | Dry plasminogen preparation |
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| WO (1) | WO1994015631A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107249622A (en) * | 2014-12-19 | 2017-10-13 | 普罗米蒂克生物治疗有限公司 | Pharmaceutical composition comprising plasminogen and its purposes |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS62153224A (en) * | 1985-12-27 | 1987-07-08 | Green Cross Corp:The | Plasminogen preparations |
| JPH0296536A (en) * | 1988-09-29 | 1990-04-09 | Green Cross Corp:The | Plasminogen dry formulation |
-
1993
- 1993-01-11 JP JP5017816A patent/JPH06211691A/en active Pending
-
1994
- 1994-01-10 WO PCT/JP1994/000019 patent/WO1994015631A1/en active Application Filing
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS62153224A (en) * | 1985-12-27 | 1987-07-08 | Green Cross Corp:The | Plasminogen preparations |
| JPH0296536A (en) * | 1988-09-29 | 1990-04-09 | Green Cross Corp:The | Plasminogen dry formulation |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107249622A (en) * | 2014-12-19 | 2017-10-13 | 普罗米蒂克生物治疗有限公司 | Pharmaceutical composition comprising plasminogen and its purposes |
| EP3233111A4 (en) * | 2014-12-19 | 2018-06-27 | Prometic Bio Therapeutics, Inc. | Pharmaceutical composition comprising plasminogen and uses thereof |
| US10441639B2 (en) | 2014-12-19 | 2019-10-15 | Prometic Biotherapeutics, Inc. | Pharmaceutical composition comprising plasminogen and uses thereof |
| TWI705822B (en) * | 2014-12-19 | 2020-10-01 | 美商波麥堤克生物治療股份有限公司 | Pharmaceutical composition comprising plasminogen and uses thereof |
| US10898553B2 (en) | 2014-12-19 | 2021-01-26 | Prometic Biotherapeutics, Inc. | Pharmaceutical composition comprising plasminogen and uses thereof |
| CN107249622B (en) * | 2014-12-19 | 2021-08-03 | 普罗米蒂克生物治疗有限公司 | Pharmaceutical composition comprising plasminogen and uses thereof |
| US11633462B2 (en) | 2014-12-19 | 2023-04-25 | Prometic Biotherapeutics, Inc. | Pharmaceutical composition comprising plasminogen and uses thereof |
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| Publication number | Publication date |
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| JPH06211691A (en) | 1994-08-02 |
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