CN1523997A - 稳定的白介素2 - Google Patents
稳定的白介素2 Download PDFInfo
- Publication number
- CN1523997A CN1523997A CNA018144454A CN01814445A CN1523997A CN 1523997 A CN1523997 A CN 1523997A CN A018144454 A CNA018144454 A CN A018144454A CN 01814445 A CN01814445 A CN 01814445A CN 1523997 A CN1523997 A CN 1523997A
- Authority
- CN
- China
- Prior art keywords
- preparation
- histidine
- stable
- sucrose
- glycine
- 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.)
- Granted
Links
- 108010002350 Interleukin-2 Proteins 0.000 title claims abstract description 95
- 102000000588 Interleukin-2 Human genes 0.000 title claims abstract description 95
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims abstract description 37
- 239000000203 mixture Substances 0.000 claims abstract description 31
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims abstract description 28
- 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 15
- 229930006000 Sucrose Natural products 0.000 claims abstract description 15
- 239000005720 sucrose Substances 0.000 claims abstract description 15
- 239000004471 Glycine Substances 0.000 claims abstract description 14
- 101001002657 Homo sapiens Interleukin-2 Proteins 0.000 claims abstract description 7
- 102000055277 human IL2 Human genes 0.000 claims abstract 2
- 238000002360 preparation method Methods 0.000 claims description 52
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 15
- 235000001014 amino acid Nutrition 0.000 claims description 10
- 150000001413 amino acids Chemical class 0.000 claims description 7
- 239000011780 sodium chloride Substances 0.000 claims description 7
- 239000007864 aqueous solution Substances 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 239000008194 pharmaceutical composition Substances 0.000 claims description 4
- 238000009472 formulation Methods 0.000 abstract description 15
- 239000000825 pharmaceutical preparation Substances 0.000 abstract description 3
- 230000035772 mutation Effects 0.000 abstract description 2
- 239000003085 diluting agent Substances 0.000 abstract 1
- 230000000087 stabilizing effect Effects 0.000 abstract 1
- 239000000872 buffer Substances 0.000 description 16
- 239000003381 stabilizer Substances 0.000 description 16
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 15
- 239000003795 chemical substances by application Substances 0.000 description 13
- 238000001556 precipitation Methods 0.000 description 10
- 235000018102 proteins Nutrition 0.000 description 10
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 9
- 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 9
- 229930195725 Mannitol Natural products 0.000 description 9
- 239000000594 mannitol Substances 0.000 description 9
- 235000010355 mannitol Nutrition 0.000 description 9
- 102000004169 proteins and genes Human genes 0.000 description 9
- 108090000623 proteins and genes Proteins 0.000 description 9
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 8
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 8
- 229940024606 amino acid Drugs 0.000 description 8
- 238000003860 storage Methods 0.000 description 8
- 150000003573 thiols Chemical class 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 6
- 229920000053 polysorbate 80 Polymers 0.000 description 6
- 239000007859 condensation product Substances 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 230000007246 mechanism Effects 0.000 description 5
- 239000000546 pharmaceutical excipient Substances 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 230000004044 response Effects 0.000 description 5
- 238000003998 size exclusion chromatography high performance liquid chromatography Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 239000004094 surface-active agent Substances 0.000 description 5
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 4
- 239000004606 Fillers/Extenders Substances 0.000 description 4
- LSDPWZHWYPCBBB-UHFFFAOYSA-N Methanethiol Chemical compound SC LSDPWZHWYPCBBB-UHFFFAOYSA-N 0.000 description 4
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 4
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 4
- 239000004141 Sodium laurylsulphate Substances 0.000 description 4
- 238000004108 freeze drying Methods 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 4
- 230000006641 stabilisation Effects 0.000 description 4
- 238000011105 stabilization Methods 0.000 description 4
- 108010088751 Albumins Proteins 0.000 description 3
- 102000009027 Albumins Human genes 0.000 description 3
- 239000004475 Arginine Substances 0.000 description 3
- 108091006905 Human Serum Albumin Proteins 0.000 description 3
- 102000008100 Human Serum Albumin Human genes 0.000 description 3
- 210000001744 T-lymphocyte Anatomy 0.000 description 3
- 108700025316 aldesleukin Proteins 0.000 description 3
- -1 amino aliphatic amino acid Chemical class 0.000 description 3
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 125000002228 disulfide group Chemical group 0.000 description 3
- HQPMKSGTIOYHJT-UHFFFAOYSA-N ethane-1,2-diol;propane-1,2-diol Chemical compound OCCO.CC(O)CO HQPMKSGTIOYHJT-UHFFFAOYSA-N 0.000 description 3
- 229920001993 poloxamer 188 Polymers 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 229940087463 proleukin Drugs 0.000 description 3
- 238000002798 spectrophotometry method Methods 0.000 description 3
- 150000005846 sugar alcohols Polymers 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- PHIQHXFUZVPYII-ZCFIWIBFSA-O (R)-carnitinium Chemical compound C[N+](C)(C)C[C@H](O)CC(O)=O PHIQHXFUZVPYII-ZCFIWIBFSA-O 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 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 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical group OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- BVHLGVCQOALMSV-JEDNCBNOSA-N L-lysine hydrochloride Chemical compound Cl.NCCCC[C@H](N)C(O)=O BVHLGVCQOALMSV-JEDNCBNOSA-N 0.000 description 2
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 2
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical group [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 2
- 230000002776 aggregation Effects 0.000 description 2
- 238000004220 aggregation Methods 0.000 description 2
- 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 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 229960004203 carnitine Drugs 0.000 description 2
- 235000018417 cysteine Nutrition 0.000 description 2
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 2
- 230000013595 glycosylation Effects 0.000 description 2
- 238000006206 glycosylation reaction Methods 0.000 description 2
- 229960005337 lysine hydrochloride Drugs 0.000 description 2
- 229930182817 methionine Natural products 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 230000035479 physiological effects, processes and functions Effects 0.000 description 2
- 229920001983 poloxamer Polymers 0.000 description 2
- 229920000136 polysorbate Polymers 0.000 description 2
- 239000001103 potassium chloride Substances 0.000 description 2
- 235000011164 potassium chloride Nutrition 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 description 1
- AGBQKNBQESQNJD-SSDOTTSWSA-N (R)-lipoic acid Chemical compound OC(=O)CCCC[C@@H]1CCSS1 AGBQKNBQESQNJD-SSDOTTSWSA-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
- 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
- DHHFDKNIEVKVKS-FMOSSLLZSA-N Betanin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC(C(=C1)O)=CC(C[C@H]2C([O-])=O)=C1[N+]2=C\C=C\1C=C(C(O)=O)N[C@H](C(O)=O)C/1 DHHFDKNIEVKVKS-FMOSSLLZSA-N 0.000 description 1
- DHHFDKNIEVKVKS-MVUYWVKGSA-N Betanin Natural products O=C(O)[C@@H]1NC(C(=O)O)=C/C(=C\C=[N+]/2\[C@@H](C(=O)[O-])Cc3c\2cc(O)c(O[C@H]2[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O2)c3)/C1 DHHFDKNIEVKVKS-MVUYWVKGSA-N 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 241001227713 Chiron Species 0.000 description 1
- 102100022641 Coagulation factor IX Human genes 0.000 description 1
- 102100026735 Coagulation factor VIII Human genes 0.000 description 1
- 241000699802 Cricetulus griseus Species 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
- 229920002307 Dextran Polymers 0.000 description 1
- 241001459693 Dipterocarpus zeylanicus Species 0.000 description 1
- 108010076282 Factor IX Proteins 0.000 description 1
- 108010054218 Factor VIII Proteins 0.000 description 1
- 102000001690 Factor VIII Human genes 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- VHOLZZKNEBBHTH-YUMQZZPRSA-N His-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@@H](N)CC1=CNC=N1 VHOLZZKNEBBHTH-YUMQZZPRSA-N 0.000 description 1
- 101000911390 Homo sapiens Coagulation factor VIII Proteins 0.000 description 1
- 208000001953 Hypotension Diseases 0.000 description 1
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 102100030703 Interleukin-22 Human genes 0.000 description 1
- 108090000978 Interleukin-4 Proteins 0.000 description 1
- 102000015696 Interleukins Human genes 0.000 description 1
- 108010063738 Interleukins Proteins 0.000 description 1
- PWKSKIMOESPYIA-BYPYZUCNSA-N L-N-acetyl-Cysteine Chemical compound CC(=O)N[C@@H](CS)C(O)=O PWKSKIMOESPYIA-BYPYZUCNSA-N 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
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 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
- 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
- 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
- 206010028980 Neoplasm Diseases 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 229920001030 Polyethylene Glycol 4000 Polymers 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 102000006010 Protein Disulfide-Isomerase Human genes 0.000 description 1
- 102000002727 Protein Tyrosine Phosphatase Human genes 0.000 description 1
- 102000016227 Protein disulphide isomerases Human genes 0.000 description 1
- 108050004742 Protein disulphide isomerases Proteins 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 206010038389 Renal cancer Diseases 0.000 description 1
- 208000006265 Renal cell carcinoma Diseases 0.000 description 1
- 108010071390 Serum Albumin Proteins 0.000 description 1
- 102000007562 Serum Albumin Human genes 0.000 description 1
- 102000013275 Somatomedins Human genes 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 241000607734 Yersinia <bacteria> Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 229960004308 acetylcysteine Drugs 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- AGBQKNBQESQNJD-UHFFFAOYSA-N alpha-Lipoic acid Natural products OC(=O)CCCCC1CCSS1 AGBQKNBQESQNJD-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000013011 aqueous formulation Substances 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 239000001654 beetroot red Substances 0.000 description 1
- 235000012677 beetroot red Nutrition 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
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 235000002185 betanin Nutrition 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 239000007979 citrate buffer Substances 0.000 description 1
- 235000015165 citric acid Nutrition 0.000 description 1
- 239000002577 cryoprotective agent Substances 0.000 description 1
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 1
- GHVNFZFCNZKVNT-UHFFFAOYSA-M decanoate Chemical compound CCCCCCCCCC([O-])=O GHVNFZFCNZKVNT-UHFFFAOYSA-M 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000009881 electrostatic interaction Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229960004222 factor ix Drugs 0.000 description 1
- 229960000301 factor viii Drugs 0.000 description 1
- FBPFZTCFMRRESA-GUCUJZIJSA-N galactitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-GUCUJZIJSA-N 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 1
- 235000003969 glutathione Nutrition 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 230000036543 hypotension Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 229960001438 immunostimulant agent Drugs 0.000 description 1
- 239000003022 immunostimulating agent Substances 0.000 description 1
- 230000003308 immunostimulating effect Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 108010074109 interleukin-22 Proteins 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- FZWBNHMXJMCXLU-BLAUPYHCSA-N isomaltotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)O1 FZWBNHMXJMCXLU-BLAUPYHCSA-N 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 235000019136 lipoic acid Nutrition 0.000 description 1
- 239000012669 liquid formulation Substances 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 210000005087 mononuclear cell Anatomy 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- RGCLLPNLLBQHPF-HJWRWDBZSA-N phosphamidon Chemical compound CCN(CC)C(=O)C(\Cl)=C(/C)OP(=O)(OC)OC RGCLLPNLLBQHPF-HJWRWDBZSA-N 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
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 108020003519 protein disulfide isomerase Proteins 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 108020000494 protein-tyrosine phosphatase Proteins 0.000 description 1
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 description 1
- 239000011677 pyridoxine Substances 0.000 description 1
- 235000008160 pyridoxine Nutrition 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 201000010174 renal carcinoma Diseases 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- FHHPUSMSKHSNKW-SMOYURAASA-M sodium deoxycholate Chemical compound [Na+].C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC([O-])=O)C)[C@@]2(C)[C@@H](O)C1 FHHPUSMSKHSNKW-SMOYURAASA-M 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 229960002663 thioctic acid Drugs 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
Images
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
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
-
- 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
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/2013—IL-2
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/04—Antineoplastic agents specific for metastasis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Epidemiology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Oncology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Train Traffic Observation, Control, And Security (AREA)
- Control Of Motors That Do Not Use Commutators (AREA)
- Sub-Exchange Stations And Push- Button Telephones (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
一种稳定的药物制剂，它含有人白细胞介素－2或其变体和稳定有效量的组氨酸。优选的的制剂包含甘氨酸和蔗糖并且IL－2变体具有一个突变，N88R。优选的制剂是冻干形式，它在用水稀释剂再配制是产生具有约5.0－6.5pH范围的溶液。
Description
技术领域
领域：本发明通常涉及药物制剂领域。更具体地说，本发明涉及稳定的、具有治疗活性、能够选择性激活T细胞(PHA-胚细胞)的白介素-2制剂并且非常优选包括表明自然杀伤(“NK”)细胞激活减少的IL-2突变蛋白质。这种稳定的具有优选特性的组合物包括下文所述的IL-2变体。
背景技术
背景：如1999年11月25日公开的相关申请PCT/US99/10643所述，白细胞介素2(IL-2)是强的免疫刺激剂，激活免疫系统的多种细胞，包括T细胞、B细胞和单核细胞。IL-2也是T细胞的强的关键的生长因子。依据这些活性的效能来试验IL-2治疗癌症的能力。人IL-2是FDA批准的用于治疗转移的肾癌和转移的黑素瘤。IL-2在合适患者中的使用因其与IL-2治疗相关的严重的毒性而受到限制，估计实际上最好只有20％的合适患者接受治疗。与IL-2治疗相关的毒性包括严重的发烧、恶心、呕吐、血管渗漏和严重的低血压。尽管有这些毒性，IL-2对于其批准的适应症是有效的。IL-2降低毒性的变体是WO99/60128申请的主题。
可获得有关稳定IL-2和其他治疗性蛋白质制剂的有意义的信息。目前批准的人IL-2制剂(ProleukinIL-2，Chiron Corporation)是冷冻干燥制剂，其包括甘露糖醇、十二烷基硫酸钠(SDS)和磷酸盐缓冲液。在下列参考文献中描述了所配制的其他治疗性蛋白质包括IL-2。
Fernandes等人，1986，由微生物产生的白介素-2的药物组合物(美国专利4,604,377)中描述了冷冻干燥的制剂，它包含稳定剂(甘露糖醇)和每毫克IL-2约100到250μg的增溶剂如十二烷基硫酸钠或者脱氧胆酸硫酸钠。相信在该参考文献中描述了目前可获得的ProleukinIL-2产品的制剂。
Patel，1994，蛋白质制剂的稳定(美国专利5358708)中描述了通过与蛋氨酸、组氨酸或其混合物混合来延长储存期的干扰素、粒细胞-巨噬细胞集落刺激因子或白介素的水性制剂。尽管以几种白介素包括白介素-2为参考，但在用IL-4进行研究中，专利权所有人发现，在所使用的稳定剂测试条件下，组氨酸稳定剂的有效性不如甲硫氨酸。
Shaked等人，1991，重组白介素-2的药物组合物及其制备方法(美国专利5,037,644)中描述了冻干或液体形式的制剂。所述制剂的赋形剂包括非离子型高分子去垢剂如吐温X405、吐温X305、PEG(4000)单硬脂酸酯，浓度大约为0.001％-5％的Tween 80和Tween 20，膨胀剂/稳定剂如蔗糖、果糖、右旋糖、麦芽糖、葡萄糖、右旋糖酐、甘露糖醇、山梨糖醇、肌醇、半乳糖醇、木糖醇、乳糖、海藻糖、人血清白蛋白和牛血清白蛋白，和浓度大约为10mM-50mM，pH范围大约为3-7的缓冲剂如甘氨酸、枸橼酸盐或磷酸盐。多元醇糖膨胀剂的浓度(wt/vol)浓度范围大约为0.025％-10％。
Roskam等人，1995，包含糖基化白介素-2的药物(美国专利5,417,970)中描述了pH6.5、包含水解明胶(或人血清白蛋白)和丙氨酸的冻干制剂。
Hora等人，1992，包含生理学上相容性稳定剂的白介素-2的药物制剂(美国专利5,078,997)中描述了液体或冻干形式的制剂。制剂可包含一种稳定剂或稳定剂的组合物，所述稳定剂如精氨酸、肉毒碱、甜菜碱、吡哆醇、聚乙烯吡咯烷酮、癸酸盐、糖、糖醇、血清白蛋白和pH为5.0-8.5的枸橼酸盐缓冲液。稳定剂的浓度如下：精氨酸为0.2-3.0％(w/v)、肉毒碱为0.2-3.0％(w/v)、蔗糖为2-6％(w/v)并且枸橼酸盐为0.01-0.3M。
Yasushi等人，1987，白介素-2和白蛋白的稳定的组合物(美国专利4,645,830)中描述了pH3-5.5、包含人清白蛋白(0.1-50mg/ml)且包含或不包含还原性赋形剂如谷胱甘肽、硫辛酸、N-乙酰基半胱氨酸或抗坏血酸(浓度0.05-20mg/ml)的稳定的水性制剂。白蛋白制剂可包含单氨基脂肪族氨基酸、环状氨基酸、单糖、糖醇或单氨基脂肪族氨基酸(浓度为5-50mg/ml)。
Lee等人，1989，在低离子强度介质氯化钠和/或氯化钾、赖氨酸盐酸盐和组氨酸中的药用血浆蛋白质制剂(美国专利4,877,608)中描述了在包含氯化钠、氯化钾或其混合物；赖氨酸盐酸盐；和组氨酸作为缓冲剂的低离子强度介质中因子VIII和其他血浆蛋白质的稳定的制剂。
Nayar，1998，稳定的低含糖量无白蛋白重组因子VIII制剂(美国专利5,763,401和5,874,408)中描述了稳定的无白蛋白含甘氨酸、组氨酸、蔗糖和NaCL的FVIII制剂。
为了尝试发现稳定的IL-2制剂(特别是WO 99/60128中优选的IL-2突变蛋白质(N88R))，我们现已找到具有生物活性的和有用的人IL-2的非常稳定的和可药用的制剂。如下所述，我们的发现基于我们相信影响稳定性的基本机理。
附图简述
图1是说明IL-2水溶液最佳pH范围的图。
图2比较了组氨酸与醋酸盐和枸橼酸盐对IL-2凝聚的稳定作用，所述凝聚是以1℃/min的速度将蛋白质溶液从25℃加热到95℃引起的。
发明概述
本发明是用组氨酸稳定的IL-2或其变体(突变蛋白质)的药物组合物或制剂。优选地，所述组合物包含在低离子强度(例如＜0.1)溶液中产生的混合物并且包含其他稳定剂如糖和氨基酸，优选蔗糖和甘氨酸。所述制剂可包含0-0.9wt.％NaCl。组合物不含白蛋白并且冻干形式的制剂可以用水快速再配制。组合物在生理上适宜的pH条件下，优选在pH大约为5.0-6.5的范围内，并且不使用表面活性剂如十二烷基硫酸钠的情况下是稳定的。重新配制的溶液近等渗的并且可通过皮下和静脉内给予。在非常优选的实施方案中，组合物中的IL-2是具有单一的氨基酸取代的突变蛋白质，优选WO 99/60128中所描述的N88R变体。
非常优选的组合物具有1-5mg/ml的变电站浓度并且包含下列水溶液形式(以wt/wt方式)：
IL-2 0.1-0.5wt％
组氨酸 0.08-1.6wt％
NaCl 0-0.9wt％
蔗糖 1-10wt％
甘氨酸 0-3wt％
pH为 5-6.5。
下文详细描述我们的制剂及其是如何发现的。
具体实施方案
本文所使用的术语IL-2包括有活性的野生型IL-2及其具有生物活性的变体或突变蛋白质如WO 99/60128中所描述的那些。在下列实施例中，我们使用称作IL-2(N88R)的IL-2，它是在氨基酸88位上的天门冬氨酸(N)突变为精氨酸(R)的人IL-2重组突变蛋白质。该突变蛋白质是由中国仓鼠卵巢(CHO)细胞表达的并且包括糖基化形式和非糖基化形式的混合物。它描述于以上所引用的作为相关申请的WO 99/60128中。
本发明的目的是识别优选的IL-2突变蛋白质冻干剂型的需要，所述剂型不含白蛋白并且具有适宜的稳定性。既然IL-2的生物测定是测定稳定性灵敏的方法，那么我们通过使用反向HPLC测定可溶性IL-2的量作为指示稳定性的测定。本文所使用的术语“稳定的”或“稳定”是指在40℃下贮存4个月后，通过反向HPLC测定的可溶性IL-2量的减少不低于原可溶性IL-2量的90％(见下表3和4)。所使用的其他指示稳定性的测定包括聚集指数，通过UV/VIS分光光度法测定聚集和通过尺寸排阻HPLC法测定可溶性聚集体。除了稳定的产品外，快速再配制(少于1分钟)的冻干物是非常优选的。最后，需要具有适宜的稳定性并且可以很容易地在生产冷冻干燥器中干燥的冻干剂型制剂。
在预制剂研究期间进行的水稳定性研究表明IL-2(N88R)在液态迅速聚集并且所述聚集是pH依赖性的。进行了两种预制剂稳定性研究：IL-2(N88R)的pH图和不同缓冲剂存在下的稳定性。这些研究的目的是识别适用于IL-2(N88R)的适宜的pH范围和适用于水稳定(减少聚集的可能性)的适宜的缓冲剂。在绘制pH图期间，制备具有不同的pH值的IL-2溶液并且在加速聚集的温度(40℃)下贮藏。在不同的时间间隔下分析样品并计算聚集速度。如图1所示，低聚集速度的最佳pH范围为pH5.0-6.5。组氨酸、醋酸盐和枸橼酸盐是在该pH范围内IL-2(N88R)的药用缓冲剂并且在浓度为20mM、包含1mg/ml IL-2(N88R)和150mM(0.9wt％)NaCl的IL-2(N88R)溶液中进行评价。将这些样品以每分钟1℃的速度从25℃加热至95℃并且通过UV分光光度计在350nm监测沉淀。如图2所示，令我们意外的是，在pH5.5时，组氨酸稳定IL-2(N88R)的作用明显地超过其他缓冲剂，这一点可通过沉淀的起始温度增加表明。在枸橼酸盐、醋酸盐和组氨酸存在下，起始温度分别为62℃、64℃和70℃。这些研究表明，对于含水条件下的IL-2来说，组氨酸不仅可用作缓冲剂，而且可用作稳定剂。
为了开发冻干剂型，研究了熟悉本领域的那些人所使用的其他赋形剂。这些赋形剂包括膨胀剂和低温保护剂如甘氨酸、蔗糖和甘露糖醇。既然聚集是在水稳定性研究时一种分子不稳定机理，那么也评价两种作为IL-2(N88R)稳定剂的表面活性剂。这些研究结果概述于下列实施例中。意外地，结果显示组氨酸对IL-2的选择性稳定作用优于其他赋形剂，如枸橼酸盐。
实施例1
在40℃下测定变体pH值的液体IL-2(N88R)的稳定性。结果显示在ph5.0-6.5时IL-2(N88R)的沉淀速度最小(图1)。因此，pH5.5选作液态制剂的最佳pH并且用于制备冻干制剂。
实施例2
我们测定不同的缓冲剂对IL-2(N88R)稳定性可能的作用。所测定的缓冲剂包括枸橼酸盐、醋酸盐和组氨酸。所使用的缓冲剂为20mM、包含1mg/ml IL-2(N88R)和150mM(0.9wt％)NaCl的IL-2(N88R)溶液。将这些稳定样品以每分钟1℃的速度从25℃加热至95℃同时通过UV/VIS分光光度计监测。与醋酸盐和枸橼酸盐相比，组氨酸通过增加沉淀温度和降低沉淀速度明显地稳定IL-2(N88R)(图2)。表1显示在这三种缓冲剂存在下IL-2(N88R)的起始沉淀温度。起始沉淀温度任意地定义为350nm下的光密度(OD350)达到某种水平(在OD350的情况下为0.2和1.0)时的温度。在组氨酸存在下，IL-2(N88R)的沉淀温度比其他缓冲剂存在下高几度。该实例证明，组氨酸除了用作液态IL-2(N88R)的缓冲剂外，可能是特异性稳定剂。
表1：IL-2(N88R)在不同缓冲剂中的沉淀温度(OD350＝0.2和OD350＝1.0)
| 缓冲剂 | 沉淀温度℃(当OD350＝0.2时) | 沉淀温度℃(当OD350＝1.0时) |
| 枸橼酸盐 | 62℃ | 64℃ |
| 醋酸盐 | 64℃ | 66℃ |
| 组氨酸 | 70℃ | 76℃ |
实施例3
为了进一步评价组氨酸的稳定作用，由不同的水溶液制剂(见表2)制备冻干的IL-2(N88R)。这些制剂的大多数都含有2wt％作为膨胀剂的甘氨酸和1wt％作为稳定剂的蔗糖。在作为对照的Proleukin(一种IL-2的商业化产品)制剂中使用5wt％的甘露糖醇。评价0.1wt％两种表面活性剂(吐温80和Pluronic F68)对表面吸附和凝聚的防止作用。所有制剂都含有组氨酸或者作为缓冲剂的枸橼酸以使pH调节到5.5。包含枸橼酸来区别组氨酸的稳定作用与pH5.5时枸橼酸的作用。在40℃温度下贮存这些冻干制剂并通过各种分析方法来进行分析，包括UV/VIS分光光度测定法、SEC-HPLC和RP-HPLC。
表3显示通过下列方法评价的各种制剂的稳定性：通过UV/VIS分光光度测定法测定凝聚、通过大小排阻HPLC(SEC-HPLC) 测定可溶解凝聚物的量并通过反向HPLC(RP-HPLC)测定蛋白质的回收百分数。样品在冻干后和在40℃加速贮存温度下贮存4个月后进行分析。冻干过程不改变冻干前IL-2(N88R)的凝聚指数，暗示IL-2(N88R)在蛋白质凝聚/沉淀方面耐受冻干过程。在40℃温度下贮存4个月后，观察到含有枸橼酸(B)、Pluronic F68(D)或甘露糖醇(E)制剂的净凝聚指数显著增加。这些结果表明在贮存期间枸橼酸或甘露糖醇、吐温80或Pluronic F68不对固体状态的IL-2(N88R)凝聚提供任何保护作用。制剂A和F的凝聚指数不显示明显的改变，暗示具有1或5mg/ml IL-2(N88R)的组氨酸、甘氨酸和蔗糖产生稳定的产品。
然而，发现在含有吐温80的制剂甚至在冻干前就有明显量的可溶性凝聚物(表3)，表明吐温80尽管可抑制不溶性凝聚物的形成，但促进可溶性IL-2(N88R)凝聚物形成。因为Pluronic F68(制剂D)也引起可溶性凝聚物明显形成，表面活性剂与IL-2(N88R)不相容。只有含2％甘氨酸、1％蔗糖和20M(0.31wt％)组氨酸(A，F)的制剂未显示所述冻干制剂在40℃下贮藏4个月后形成可检测到的可溶性聚集体，再次暗示IL-2(N88R)可由组氨酸稳定。
通过RP-HPLC测定冻干和贮藏后IL-2(N88R)的总回收率(表3)。除了含甘露糖醇的制剂外，所有制剂冻干后IL-2(N88R)的回收率大于大约96％。这些制剂在40℃下贮藏4个月后，在含2％甘氨酸、1％蔗糖和20mM(0.31wt％)组氨酸并且分别含1和5mg/ml IL-2(N88R)的制剂A和F中IL-2(N88R)的回收率大约为92％。这些数据(制剂A和F中IL-2(N88R)的回收率大于90％)再次暗示组氨酸稳定IL-2(N88R)，而表面活性剂不能稳定蛋白质。
表2：冻干IL-2(N88R)制剂的组成
| 制剂ID | A | B | C | D | E | F |
| IL-2(N88R)，mg/ml | 1 | 1 | 1 | 1 | 1 | 5 |
| 甘氨酸，wt％(w/w) | 2 | 2 | 2 | 2 | 0 | 2 |
| 蔗糖，wt％(w/w) | 1 | 1 | 1 | 1 | 0 | 1 |
| 甘露糖醇，wt％(w/w) | 0 | 0 | 0 | 0 | 5 | 0 |
| 枸橼酸钠，wt％(w/w) | 0 | 0.6 | 0 | 0 | 0 | 0 |
| 组氨酸wt％(w/w) | 0.31 | 0 | 0.31 | 0.31 | 0.31 | 0.31 |
| 吐温80，wt％(w/w) | 0 | 0 | 0.1 | 0 | 0 | 0 |
| Pluronic F68，wt％(w/w) | 0 | 0 | 0 | 0.1 | 0 | 0 |
| pH | 5.5 | 5.5 | 5.5 | 5.5 | 5.5 | 5.5 |
表3：IL-2(N88R)在冻干过程中和冻干制剂在40℃下贮藏4个月期间的稳定性
| 制剂ID | A | B | C | D | E | F |
| 聚集指标(％) | ||||||
| 冻干前 | 4.4 | 5.2 | 1.5 | 4.7 | 5.6 | 1.6 |
| 冻干后 | 5.2 | 5.1 | 0.7 | 1.5 | 3.8 | 1.6 |
| 在40℃下4个月 | 2.9 | 13.9 | 2.4 | 14.3 | 20.8 | 3.3 |
| 通过SEC-HPLC测定可溶性聚集体(％) | ||||||
| 冻干前 | NDa | ND | 4.4％ | ND | ND | NAb |
| 冻干后 | ND | ND | 7.2％ | ND | ND | NA |
| 在40℃下4个月 | ND | 4.2％ | 32.9％ | 15.9％ | 12.2％ | NA |
| 通过RP-HPLC测定回收率(％) | ||||||
| 冻干前 | 100 | 100 | 100 | 100 | 100 | 100 |
| 冻干后 | 96.5 | 95.7 | 96.4 | 99.4 | 91.7 | 97.8 |
| 在40℃下4个月 | 92.5 | 82.9 | 71.9 | 84.3 | 79.1 | 91.7 |
aND＝不可检测的
bNA＝不可获得的
实施例4
也将与制剂A具有相同组成的野生型IL-2的水溶液制剂冻干。比较野生型IL-2与IL-2(N88R)在40℃下的稳定性(表4)。
表4；野生型IL-2(N88R)制剂在冻干过程中和冻干制剂在储存期间的稳定性
| 条件 | 聚集指标(％) | 可溶性聚集体(％) | 回收率(％) |
| 冻干前 | 4.2 | NDa | 100 |
| 冻干后 | 5.1 | ND | 93.6 |
| 在40℃下4个月 | 4.3 | ND | 93.0 |
aND＝不可检测的
讨论
人IL-2具有133个氨基酸，它们形成六种螺旋结构(A-F)。这些螺旋中的四种形成称作四-螺旋束的基元。Cys58和cys105之间的分子内二硫键位于螺旋间延长的环状结构上。游离cys125位于结合氨基酸117-133的螺旋F上。
对组氨酸是IL-2特异性稳定剂的惊奇发现暗示组氨酸可能与IL-2以特异的方式相互作用，由此稳定水溶液状态和冻干状态的分子。IL-2不稳定的主要机制之一是凝聚，这是由于硫醇-二硫化物的交换反应形成低聚物所致。因此，一种假设是事实上组氨酸可抑制或减少IL-2中硫醇-二硫化物的交换反应。由于野生型IL-2、IL-2(N88R)和可能的其他IL-2变体分子具有一个二硫键和游离的半胱氨酸(cys125)，cys125上的游离-SH基团易于通过硫醇/二硫化物交换途径与二硫键进行反应，由此导致凝聚/沉淀发生。
Bulaj等人最近描述了硫醇/二硫化物交换机制(多肽中半胱氨酸硫醇的离子化-反应活性的关系，Biochemistry 1998，6，23；37(25)：8965-72)。在模型肽和蛋白质的研究中，显示反应速率对静电力以及蛋白质的二级结构是敏感的。硫醇中硫原子周围的电子分布可受改变硫醇pKa的附近电荷和整个键诱导作用存在的影响。例如，硫醇/二硫化物中硫醇反应活性增加归因于其pKa因附近正电荷或者来自附近α螺旋结构的肽偶极分布的降低。相反地，巯基附近的负电荷可使硫醇的pKa升高并导致硫醇/二硫化物反应速率的降低。所提出的机制也得到其他研究的支持，已经表明通过邻接正电荷残基稳定蛋白酪氨酸磷酸酯酶(Zhang和Dixon，1993 Active site labeling ofthe Yersinia protein tyrosine phosphatase：the determination ofthe pKa of the active site cysteine and the function of theconserved histidine 402.Biochemistry 1993 Sep 14；32(36)：9340-5)和蛋白二硫化物异构酶(Kortemme等人，Electrostatic interactions in the active site of the N-terminalthioredoxin-like domain of protein disulfide isomerase.Biochemistry 1996 Nov 19；35(46)：14503-11)中高反应活性的硫醇盐离子。
基于硫醇/二硫化物交换反应的这些特性，可以预想一种稳定机制，那就是组氨酸在IL-2(N88R)的稳定中起动力学或者热力学作用。在二硫化物桥Cys58-Cys105附近有5个谷氨酸残基(57，60，61，62和106)。组氨酸和这些负电荷残基的特异性结合可导致硫醇/二硫化物交换反应的动力学屏障或者二硫化物桥附近组氨酸的结合可在热力学上将IL-2分子稳定为不易于进行凝聚反应的构型。另外，His-Glu离子的相互作用也可产生立体上的障碍，这将进一步使硫醇/二硫化物交换中决定速率的步骤难以进行，其中在三个参与的硫原子之间形成中间体过渡状态。组氨酸易于接近谷氨酸残基是非常可能的，因为二硫化物位于蛋白质表面的延长的环结构上。
给出了以上实施例，预计本领域技术人员可对本文所公开的发明进行改变。因此，希望上述实施例仅仅是说明性的并且本文公开的发明将仅通过下列权利要求来限定。
Claims (10)
1.一种稳定的药物组合物，包含用组氨酸稳定的人白介素-2或其变体。
2.冻干形式的权利要求1的制剂，它可以在水中迅速再配制。
3.水溶液形式的权利要求1的制剂，具有约5.0-6.5的pH范围。
4.权利要求1的制剂，其中包含甘氨酸。
5.权利要求1的制剂，其中包含蔗糖。
6.权利要求1的制剂，其中包含甘氨酸和蔗糖。
7.权利要求1的制剂，其中的IL-2是突变蛋白质。
8.权利要求7的制剂，其中IL-2是具有一个氨基酸取代基N88R的突变蛋白质。
9.一种稳定的冻干组合物，在用水再配制前，它含有下列组分：
IL-2 0.1-5mg/ml
组氨酸 0.08-1.6wt％
NaCl 0-0.9wt％
蔗糖 1-10wt％和
甘氨酸 0-3wt％
pH 5-6.5
10.权利要求9的制剂，其中IL-2是具有一个氨基酸取代基N88R的突变蛋白质。
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US09/605,577 | 2000-06-28 | ||
| US09/605,577 US6689353B1 (en) | 2000-06-28 | 2000-06-28 | Stabilized interleukin 2 |
| PCT/US2001/020675 WO2002000243A2 (en) | 2000-06-28 | 2001-06-27 | Stabilized interleukin 2 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1523997A true CN1523997A (zh) | 2004-08-25 |
| CN1523997B CN1523997B (zh) | 2010-09-01 |
Family
ID=24424265
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN018144454A Expired - Fee Related CN1523997B (zh) | 2000-06-28 | 2001-06-27 | 稳定的白介素2 |
Country Status (24)
| Country | Link |
|---|---|
| US (1) | US6689353B1 (zh) |
| EP (1) | EP1370280B1 (zh) |
| JP (3) | JP5522878B2 (zh) |
| KR (1) | KR100799402B1 (zh) |
| CN (1) | CN1523997B (zh) |
| AR (1) | AR029139A1 (zh) |
| AU (1) | AU2001273063A1 (zh) |
| BR (1) | BR0112101B1 (zh) |
| CA (1) | CA2413334C (zh) |
| CO (1) | CO5290308A1 (zh) |
| CU (1) | CU23536A3 (zh) |
| DE (1) | DE60142412D1 (zh) |
| DO (1) | DOP2001000197A (zh) |
| EC (1) | ECSP014106A (zh) |
| ES (1) | ES2344729T3 (zh) |
| HN (1) | HN2001000139A (zh) |
| IL (2) | IL153587A0 (zh) |
| MX (1) | MXPA03000046A (zh) |
| MY (1) | MY128629A (zh) |
| PE (1) | PE20020127A1 (zh) |
| SV (1) | SV2002000512A (zh) |
| TW (1) | TWI235063B (zh) |
| UY (1) | UY26805A1 (zh) |
| WO (1) | WO2002000243A2 (zh) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103063829A (zh) * | 2012-12-21 | 2013-04-24 | 杭州茂天赛科技有限公司 | 一种冻存液 |
| WO2022100684A1 (zh) * | 2020-11-13 | 2022-05-19 | 江苏恒瑞医药股份有限公司 | 一种包含人白细胞介素2变体或其衍生物的药物组合物及其用途 |
Families Citing this family (59)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8531276B2 (en) * | 2000-03-15 | 2013-09-10 | Logitech Europe S.A. | State-based remote control system |
| US6689353B1 (en) * | 2000-06-28 | 2004-02-10 | Bayer Pharmaceuticals Corporation | Stabilized interleukin 2 |
| US7407955B2 (en) | 2002-08-21 | 2008-08-05 | Boehringer Ingelheim Pharma Gmbh & Co., Kg | 8-[3-amino-piperidin-1-yl]-xanthines, the preparation thereof and their use as pharmaceutical compositions |
| JP5057648B2 (ja) * | 2002-09-26 | 2012-10-24 | 塩野義製薬株式会社 | 安定化されたタンパク組成物 |
| NZ550119A (en) | 2004-04-08 | 2009-10-30 | Biomatrica Inc | Method for storing biological material in a dissolvable matrix |
| US20060099567A1 (en) * | 2004-04-08 | 2006-05-11 | Biomatrica, Inc. | Integration of sample storage and sample management for life science |
| US7718622B2 (en) * | 2005-03-04 | 2010-05-18 | Dynavax Technologies Corporation | Compositions comprising structurally stable conjugate molecules |
| JP2009529542A (ja) * | 2006-03-10 | 2009-08-20 | ダイアックス コーポレーション | エカランチドに関する配合物 |
| CN102838599A (zh) | 2006-05-04 | 2012-12-26 | 贝林格尔.英格海姆国际有限公司 | 多晶型 |
| PE20080251A1 (es) | 2006-05-04 | 2008-04-25 | Boehringer Ingelheim Int | Usos de inhibidores de dpp iv |
| EP1852108A1 (en) | 2006-05-04 | 2007-11-07 | Boehringer Ingelheim Pharma GmbH & Co.KG | DPP IV inhibitor formulations |
| AU2008260483A1 (en) * | 2007-06-01 | 2008-12-11 | Acologix, Inc. | High temperature stable peptide formulation |
| PE20140960A1 (es) | 2008-04-03 | 2014-08-15 | Boehringer Ingelheim Int | Formulaciones que comprenden un inhibidor de dpp4 |
| DE102008023820A1 (de) | 2008-05-08 | 2009-11-12 | Aicuris Gmbh & Co. Kg | Mittel zur Behandlung und/oder Prophylaxe einer Autoimmunerkrankung und zur Bildung von Regulatorischen T-Zellen |
| BRPI0916997A2 (pt) | 2008-08-06 | 2020-12-15 | Boehringer Ingelheim International Gmbh | Inibidor de dpp-4 e seu uso |
| US20200155558A1 (en) | 2018-11-20 | 2020-05-21 | Boehringer Ingelheim International Gmbh | Treatment for diabetes in patients with insufficient glycemic control despite therapy with an oral antidiabetic drug |
| HUE037449T2 (hu) | 2008-10-17 | 2018-08-28 | Sanofi Aventis Deutschland | Egy inzulin és egy GLP-1 agonista kombinációja |
| US8519125B2 (en) * | 2009-05-11 | 2013-08-27 | Biomatrica, Inc. | Compositions and methods for biological sample storage |
| AR080669A1 (es) | 2009-11-13 | 2012-05-02 | Sanofi Aventis Deutschland | Composicion farmaceutica que comprende un agonista de glp-1, una insulina y metionina |
| WO2011058082A1 (de) * | 2009-11-13 | 2011-05-19 | Sanofi-Aventis Deutschland Gmbh | Pharmazeutische zusammensetzung umfassend einen glp-1-agonisten und methionin |
| KR102668834B1 (ko) | 2009-11-27 | 2024-05-24 | 베링거 인겔하임 인터내셔날 게엠베하 | 리나글립틴과 같은 ｄｐｐ－ｉｖ 억제제를 사용한 유전자형 검사된 당뇨병 환자의 치료 |
| ES2935300T3 (es) | 2010-05-05 | 2023-03-03 | Boehringer Ingelheim Int | Combiterapia |
| WO2012018638A2 (en) | 2010-07-26 | 2012-02-09 | Biomatrica, Inc. | Compositions for stabilizing dna, rna and proteins in blood and other biological samples during shipping and storage at ambient temperatures |
| WO2012018639A2 (en) | 2010-07-26 | 2012-02-09 | Biomatrica, Inc. | Compositions for stabilizing dna, rna and proteins in saliva and other biological samples during shipping and storage at ambient temperatures |
| ES2606554T3 (es) | 2010-08-30 | 2017-03-24 | Sanofi-Aventis Deutschland Gmbh | Uso de AVE0010 para la fabricación de un medicamento para el tratamiento de la diabetes mellitus de tipo 2 |
| CA3144697A1 (en) | 2010-11-12 | 2012-05-18 | Nektar Therapeutics | Conjugates of an il-2 moiety and a polymer |
| US9034883B2 (en) | 2010-11-15 | 2015-05-19 | Boehringer Ingelheim International Gmbh | Vasoprotective and cardioprotective antidiabetic therapy |
| JP6034798B2 (ja) * | 2010-12-02 | 2016-11-30 | オンコリティクス バイオテク，インコーポレーテッド | 液体ウイルス製剤 |
| US9821032B2 (en) | 2011-05-13 | 2017-11-21 | Sanofi-Aventis Deutschland Gmbh | Pharmaceutical combination for improving glycemic control as add-on therapy to basal insulin |
| BR112014004726A2 (pt) | 2011-08-29 | 2017-04-04 | Sanofi Aventis Deutschland | combinação farmacêutica para uso no controle glicêmico em pacientes de diabetes tipo 2 |
| TWI559929B (en) | 2011-09-01 | 2016-12-01 | Sanofi Aventis Deutschland | Pharmaceutical composition for use in the treatment of a neurodegenerative disease |
| WO2013171167A1 (en) | 2012-05-14 | 2013-11-21 | Boehringer Ingelheim International Gmbh | A xanthine derivative as dpp -4 inhibitor for use in the treatment of podocytes related disorders and/or nephrotic syndrome |
| EP3249054A1 (en) | 2012-12-20 | 2017-11-29 | Biomatrica, INC. | Formulations and methods for stabilizing pcr reagents |
| GB201403775D0 (en) | 2014-03-04 | 2014-04-16 | Kymab Ltd | Antibodies, uses & methods |
| ES2891555T3 (es) | 2014-06-10 | 2022-01-28 | Biomatrica Inc | Estabilización de trombocitos a temperaturas ambiente |
| EP3482766B1 (en) | 2014-08-11 | 2020-05-20 | Delinia, Inc. | Modified il-2 variants that selectively activate regulatory t cells for the treatment of autoimmune diseases |
| PL3229828T3 (pl) | 2014-12-12 | 2023-07-31 | Sanofi-Aventis Deutschland Gmbh | Formulacja o ustalonym stosunku insuliny glargine/liksysenatydu |
| TWI748945B (zh) | 2015-03-13 | 2021-12-11 | 德商賽諾菲阿凡提斯德意志有限公司 | 第２型糖尿病病患治療 |
| TW201705975A (zh) | 2015-03-18 | 2017-02-16 | 賽諾菲阿凡提斯德意志有限公司 | 第２型糖尿病病患之治療 |
| EP4242628A3 (en) | 2015-12-08 | 2023-11-08 | Biomatrica, INC. | Reduction of erythrocyte sedimentation rate |
| US20170204154A1 (en) | 2016-01-20 | 2017-07-20 | Delinia, Inc. | Molecules that selectively activate regulatory t cells for the treatment of autoimmune diseases |
| EP4233840A3 (en) | 2016-06-10 | 2023-10-18 | Boehringer Ingelheim International GmbH | Combinations of linagliptin and metformin |
| US9567399B1 (en) | 2016-06-20 | 2017-02-14 | Kymab Limited | Antibodies and immunocytokines |
| EP3534947A1 (en) | 2016-11-03 | 2019-09-11 | Kymab Limited | Antibodies, combinations comprising antibodies, biomarkers, uses & methods |
| CN110167957A (zh) | 2016-11-08 | 2019-08-23 | 德里尼亚公司 | 用于治疗自身免疫疾病的il-2变体 |
| WO2018170288A1 (en) | 2017-03-15 | 2018-09-20 | Pandion Therapeutics, Inc. | Targeted immunotolerance |
| CN111010866A (zh) | 2017-05-24 | 2020-04-14 | 潘迪恩治疗公司 | 靶向免疫耐受性 |
| AU2018372167B2 (en) | 2017-11-21 | 2023-12-07 | The Board Of Trustees Of The Leland Stanford Junior University | Partial agonists of interleukin-2 |
| US10174092B1 (en) | 2017-12-06 | 2019-01-08 | Pandion Therapeutics, Inc. | IL-2 muteins |
| US10946068B2 (en) | 2017-12-06 | 2021-03-16 | Pandion Operations, Inc. | IL-2 muteins and uses thereof |
| MX2020009857A (es) | 2018-03-28 | 2021-01-08 | Ascendis Pharma Oncology Div A/S | Conjugados de interleucina-2 (il-2). |
| JP2022533702A (ja) | 2019-05-20 | 2022-07-25 | パンディオン・オペレーションズ・インコーポレイテッド | ＭＡｄＣＡＭ標的化免疫寛容 |
| KR20220114595A (ko) | 2019-12-17 | 2022-08-17 | 암젠 인크 | 치료에서의 사용을 위한 이중 인터류킨-2/tnf 수용체 효현제 |
| JP2023510115A (ja) | 2019-12-20 | 2023-03-13 | リジェネロン・ファーマシューティカルズ・インコーポレイテッド | 新規ｉｌ２アゴニストおよびそれらの使用方法 |
| EP4087865A2 (en) | 2020-01-10 | 2022-11-16 | Bright Peak Therapeutics AG | Modified il-2 polypeptides and uses thereof |
| JP7303391B2 (ja) | 2020-01-14 | 2023-07-04 | シンセカイン インコーポレイテッド | バイアス型ｉｌ２ムテイン、方法、および組成物 |
| WO2021168079A1 (en) | 2020-02-21 | 2021-08-26 | Pandion Operations, Inc. | Tissue targeted immunotolerance with a cd39 effector |
| BR112022022826A2 (pt) | 2020-06-03 | 2022-12-13 | Ascendis Pharma Oncology Div A/S | Sequências de il-2 e usos das mesmas |
| TWI815194B (zh) | 2020-10-22 | 2023-09-11 | 美商基利科學股份有限公司 | 介白素2-Fc融合蛋白及使用方法 |
Family Cites Families (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS60215631A (ja) * | 1984-04-09 | 1985-10-29 | Takeda Chem Ind Ltd | インタ−ロイキン−２組成物 |
| DE3583880D1 (de) * | 1984-04-09 | 1991-10-02 | Takeda Chemical Industries Ltd | Stabile interleukin-2-zusammensetzung. |
| JPH0645551B2 (ja) * | 1986-01-07 | 1994-06-15 | 塩野義製薬株式会社 | インタ−ロイキン−２組成物 |
| JPH0229016A (ja) * | 1989-06-09 | 1990-01-31 | Alpine Electron Inc | イコライザ装置 |
| US5358708A (en) | 1993-01-29 | 1994-10-25 | Schering Corporation | Stabilization of protein formulations |
| US5656730A (en) * | 1995-04-07 | 1997-08-12 | Enzon, Inc. | Stabilized monomeric protein compositions |
| US6267958B1 (en) * | 1995-07-27 | 2001-07-31 | Genentech, Inc. | Protein formulation |
| US5763401A (en) | 1996-07-12 | 1998-06-09 | Bayer Corporation | Stabilized albumin-free recombinant factor VIII preparation having a low sugar content |
| AU8985898A (en) * | 1997-07-31 | 1999-02-22 | Rhodia Chimie | Cosmetic composition comprising a functionalised polyorganosiloxane |
| DZ2788A1 (fr) * | 1998-05-15 | 2003-12-01 | Bayer Ag | Agonistes et antagonistes selectifs à IL-2. |
| US6689353B1 (en) * | 2000-06-28 | 2004-02-10 | Bayer Pharmaceuticals Corporation | Stabilized interleukin 2 |
-
2000
- 2000-06-28 US US09/605,577 patent/US6689353B1/en not_active Expired - Lifetime
-
2001
- 2001-06-14 AR ARP010102841A patent/AR029139A1/es active IP Right Grant
- 2001-06-26 DO DO2001000197A patent/DOP2001000197A/es unknown
- 2001-06-26 MY MYPI20013006A patent/MY128629A/en unknown
- 2001-06-26 CO CO01050546A patent/CO5290308A1/es not_active Application Discontinuation
- 2001-06-27 EC EC2001004106A patent/ECSP014106A/es unknown
- 2001-06-27 CA CA2413334A patent/CA2413334C/en not_active Expired - Lifetime
- 2001-06-27 SV SV2001000512A patent/SV2002000512A/es unknown
- 2001-06-27 DE DE60142412T patent/DE60142412D1/de not_active Expired - Lifetime
- 2001-06-27 EP EP01952296A patent/EP1370280B1/en not_active Expired - Lifetime
- 2001-06-27 AU AU2001273063A patent/AU2001273063A1/en not_active Abandoned
- 2001-06-27 JP JP2002505024A patent/JP5522878B2/ja not_active Expired - Lifetime
- 2001-06-27 ES ES01952296T patent/ES2344729T3/es not_active Expired - Lifetime
- 2001-06-27 UY UY26805A patent/UY26805A1/es not_active IP Right Cessation
- 2001-06-27 WO PCT/US2001/020675 patent/WO2002000243A2/en active Application Filing
- 2001-06-27 IL IL15358701A patent/IL153587A0/xx active IP Right Grant
- 2001-06-27 KR KR1020027017859A patent/KR100799402B1/ko active IP Right Grant
- 2001-06-27 CN CN018144454A patent/CN1523997B/zh not_active Expired - Fee Related
- 2001-06-27 MX MXPA03000046A patent/MXPA03000046A/es active IP Right Grant
- 2001-06-27 BR BRPI0112101A patent/BR0112101B1/pt not_active IP Right Cessation
- 2001-06-28 HN HN2001000139A patent/HN2001000139A/es unknown
- 2001-06-28 TW TW090115670A patent/TWI235063B/zh not_active IP Right Cessation
- 2001-06-28 PE PE2001000639A patent/PE20020127A1/es not_active Application Discontinuation
-
2002
- 2002-12-23 IL IL153587A patent/IL153587A/en unknown
- 2002-12-24 CU CU20020334A patent/CU23536A3/es not_active IP Right Cessation
-
2012
- 2012-06-21 JP JP2012139913A patent/JP2012211166A/ja active Pending
-
2014
- 2014-09-08 JP JP2014182175A patent/JP5989728B2/ja not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103063829A (zh) * | 2012-12-21 | 2013-04-24 | 杭州茂天赛科技有限公司 | 一种冻存液 |
| CN103063829B (zh) * | 2012-12-21 | 2015-01-14 | 杭州茂天赛科技有限公司 | 一种冻存液 |
| WO2022100684A1 (zh) * | 2020-11-13 | 2022-05-19 | 江苏恒瑞医药股份有限公司 | 一种包含人白细胞介素2变体或其衍生物的药物组合物及其用途 |
Also Published As
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1523997B (zh) | 稳定的白介素2 | |
| KR100656125B1 (ko) | 장기 안정화 제제 | |
| ES2381674T3 (es) | Método de estabilizar proteínas | |
| TWI409082B (zh) | 用於長效型粒細胞集落刺激因子（ｇ－ｃｓｆ）共軛物之液態劑型 | |
| US7731948B2 (en) | Stabilized interferon liquid formulations | |
| US6250469B1 (en) | Formulations for protection of peg-interferon alpha conjugates | |
| WO1990000397A1 (en) | A pharmaceutical composition for interleukin-2 containing physiologically compatible stabilizers | |
| US20050163752A1 (en) | Human interferon-beta formulations | |
| CN1191863C (zh) | 用于保护PEG－α干扰素缀合物的制剂 | |
| AU2013336206A1 (en) | Stable pharmaceutical composition of peginterferon alpha-2b | |
| WO2008066322A1 (en) | A stable hsa-free and antioxidant-free pharmaceutical composition comprising interferon-beta |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| ASS | Succession or assignment of patent right |
Owner name: AI LIMITED KUMARIS KG CO., LTD. Free format text: FORMER OWNER: BAYER CORP., USA Effective date: 20070525 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20070525 Address after: Germany Wuppertal Applicant after: Aicuris GmbH & Co. KG Address before: American Pennsylvania Applicant before: Miles Inc. |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20100901 Termination date: 20200627 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |