JP5914363B2 - 低減されたvwf結合を有する因子viii分子 - Google Patents
低減されたvwf結合を有する因子viii分子 Download PDFInfo
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- JP5914363B2 JP5914363B2 JP2012553254A JP2012553254A JP5914363B2 JP 5914363 B2 JP5914363 B2 JP 5914363B2 JP 2012553254 A JP2012553254 A JP 2012553254A JP 2012553254 A JP2012553254 A JP 2012553254A JP 5914363 B2 JP5914363 B2 JP 5914363B2
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- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
- 229960003299 ketamine Drugs 0.000 description 1
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- 125000005439 maleimidyl group Chemical group C1(C=CC(N1*)=O)=O 0.000 description 1
- 239000011565 manganese chloride Substances 0.000 description 1
- 235000002867 manganese chloride Nutrition 0.000 description 1
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- 229960001412 pentobarbital Drugs 0.000 description 1
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- 125000000951 phenoxy group Chemical group [H]C1=C([H])C([H])=C(O*)C([H])=C1[H] 0.000 description 1
- 125000000843 phenylene group Chemical group C1(=C(C=CC=C1)*)* 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
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- PYJNAPOPMIJKJZ-UHFFFAOYSA-N phosphorylcholine chloride Chemical compound [Cl-].C[N+](C)(C)CCOP(O)(O)=O PYJNAPOPMIJKJZ-UHFFFAOYSA-N 0.000 description 1
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- 229920002627 poly(phosphazenes) Polymers 0.000 description 1
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- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
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- 238000002360 preparation method Methods 0.000 description 1
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- 238000012545 processing Methods 0.000 description 1
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- 238000011321 prophylaxis Methods 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 235000019423 pullulan Nutrition 0.000 description 1
- 238000010188 recombinant method Methods 0.000 description 1
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- 230000000284 resting effect Effects 0.000 description 1
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- 150000003839 salts Chemical class 0.000 description 1
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- 238000013207 serial dilution Methods 0.000 description 1
- 230000002226 simultaneous effect Effects 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 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
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
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- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
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- 239000006228 supernatant Substances 0.000 description 1
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- 230000003612 virological effect Effects 0.000 description 1
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- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 description 1
Classifications
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- A61K47/6835—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
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- A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/745—Blood coagulation or fibrinolysis factors
- C07K14/755—Factors VIII, e.g. factor VIII C (AHF), factor VIII Ag (VWF)
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/30—Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
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- C07K2319/31—Fusion polypeptide fusions, other than Fc, for prolonged plasma life, e.g. albumin
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Immunology (AREA)
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- Hematology (AREA)
- Molecular Biology (AREA)
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- Zoology (AREA)
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- Biophysics (AREA)
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- Microbiology (AREA)
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- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Diabetes (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicinal Preparation (AREA)
Description
フォンウィレブランド因子(vWF): vWFは血漿中に存在する大きな単量体/多量体の糖タンパク質であり、内皮(Weibel-Palade体)、巨核球(血小板のα-顆粒)および内皮下結合組織で構成的に生成される。その一次機能は他のタンパク質、特に因子VIIIへの結合であり、それは創傷部位への血小板粘着で重要である。
ATRRYYLGAVELSWDYMQSDLGELPVDARFPPRVPKSFPFNTSVVYKKTLFVEFTDHLFNIAKPRPPWMGLLGPTIQAEVYDTVVITLKNMASHPVSLHAVGVSYWKASEGAEYDDQTSQREKEDDKVFPGGSHTYVWQVLKENGPMASDPLCLTYSYLSHVDLVKDLNSGLIGALLVCREGSLAKEKTQTLHKFILLFAVFDEGKSWHSETKNSLMQDRDAASARAWPKMHTVNGYVNRSLPGLIGCHRKSVYWHVIGMGTTPEVHSIFLEGHTFLVRNHRQASLEISPITFLTAQTLLMDLGQFLLFCHISSHQHDGMEAYVKVDSCPEEPQLRMKNNEEAEDYDDDLTDSEMDVVRFDDDNSPSFIQIRSVAKKHPKTWVHYIAAEEEDWDYAPLVLAPDDRSYKSQYLNNGPQRIGRKYKKVRFMAYTDETFKTREAIQHESGILGPLLYGEVGDTLLIIFKNQASRPYNIYPHGITDVRPLYSRRLPKGVKHLKDFPILPGEIFKYKWTVTVEDGPTKSDPRCLTRYYSSFVNMERDLASGLIGPLLICYKESVDQRGNQIMSDKRNVILFSVFDENRSWYLTENIQRFLPNPAGVQLEDPEFQASNIMHSINGYVFDSLQLSVCLHEVAYWYILSIGAQTDFLSVFFSGYTFKHKMVYEDTLTLFPFSGETVFMSMENPGLWILGCHNSDFRNRGMTALLKVSSCDKNTGDYYEDSYEDISAYLLSKNNAIEPRSFSQNSRHPSTRQKQFNATTIPENDIEKTDPWFAHRTPMPKIQNVSSSDLLMLLRQSPTPHGLSLSDLQEAKYETFSDDPSPGAIDSNNSLSEMTHFRPQLHHSGDMVFTPESGLQLRLNEKLGTTAATELKKLDFKVSSTSNNLISTIPSDNLAAGTDNTSSLGPPSMPVHYDSQLDTTLFGKKSSPLTESGGPLSLSEENNDSKLLESGLMNSQESSWGKNVSSTESGRLFKGKRAHGPALLTKDNALFKVSISLLKTNKTSNNSATNRKTHIDGPSLLIENSPSVWQNILESDTEFKKVTPLIHDRMLMDKNATALRLNHMSNKTTSSKNMEMVQQKKEGPIPPDAQNPDMSFFKMLFLPESARWIQRTHGKNSLNSGQGPSPKQLVSLGPEKSVEGQNFLSEKNKVVVGKGEFTKDVGLKEMVFPSSRNLFLTNLDNLHENNTHNQEKKIQEEIEKKETLIQENVVLPQIHTVTGTKNFMKNLFLLSTRQNVEGSYDGAYAPVLQDFRSLNDSTNRTKKHTAHFSKKGEEENLEGLGNQTKQIVEKYACTTRISPNTSQQNFVTQRSKRALKQFRLPLEETELEKRIIVDDTSTQWSKNMKHLTPSTLTQIDYNEKEKGAITQSPLSDCLTRSHSIPQANRSPLPIAKVSSFPSIRPIYLTRVLFQDNSSHLPAASYRKKDSGVQESSHFLQGAKKNNLSLAILTLEMTGDQREVGSLGTSATNSVTYKKVENTVLPKPDLPKTSGKVELLPKVHIYQKDLFPTETSNGSPGHLDLVEGSLLQGTEGAIKWNEANRPGKVPFLRVATESSAKTPSKLLDPLAWDNHYGTQIPKEEWKSQEKSPEKTAFKKKDTILSLNACESNHAIAAINEGQNKPEIEVTWAKQGRTERLCSQNPPVLKRHQREITRTTLQSDQEEIDYDDTISVEMKKEDFDIYDEDENQSPRSFQKKTRHYFIAAVERLWDYGMSSSPHVLRNRAQSGSVPQFKKVVFQEFTDGSFTQPLYRGELNEHLGLLGPYIRAEVEDNIMVTFRNQASRPYSFYSSLISYEEDQRQGAEPRKNFVKPNETKTYFWKVQHHMAPTKDEFDCKAWAYFSDVDLEKDVHSGLIGPLLVCHTNTLNPAHGRQVTVQEFALFFTIFDETKSWYFTENMERNCRAPCNIQMEDPTFKENYRFHAINGYIMDTLPGLVMAQDQRIRWYLLSMGSNENIHSIHFSGHVFTVRKKEEYKMALYNLYPGVFETVEMLPSKAGIWRVECLIGEHLHAGMSTLFLVYSNKCQTPLGMASGHIRDFQITASGQYGQWAPKLARLHYSGSINAWSTKEPFSWIKVDLLAPMIIHGIKTQGARQKFSSLYISQFIIMYSLDGKKWQTYRGNSTGTLMVFFGNVDSSGIKHNIFNPPIIARYIRLHPTHYSIRSTLRMELMGCDLNSCSMPLGMESKAISDAQITASSYFTNMFATWSPSKARLHLQGRSNAWRPQVNNPKEWLQVDFQKTMKVTGVTTQGVKSLLTSMYVKEFLISSSQDGHQWTLFFQNGKVKVFQGNQDSFTPVVNSLDPPLLTRYLRIHPQSWVHQIALRMEVLGCEAQDLY
ATRRYYLGAVELSWDYMQSDLGELPVDARFPPRVPKSFPFNTSVVYKKTLFVEFTDHLFNIAKPRPPWMGLLGPTIQAEVYDTVVITLKNMASHPVSLHAVGVSYWKASEGAEYDDQTSQREKEDDKVFPGGSHTYVWQVLKENGPMASDPLCLTYSYLSHVDLVKDLNSGLIGALLVCREGSLAKEKTQTLHKFILLFAVFDEGKSWHSETKNSLMQDRDAASARAWPKMHTVNGYVNRSLPGLIGCHRKSVYWHVIGMGTTPEVHSIFLEGHTFLVRNHRQASLEISPITFLTAQTLLMDLGQFLLFCHISSHQHDGMEAYVKVDSCPEEPQLRMKNNEEAEDYDDDLTDSEMDVVRFDDDNSPSFIQIRSVAKKHPKTWVHYIAAEEEDWDYAPLVLAPDDRSYKSQYLNNGPQRIGRKYKKVRFMAYTDETFKTREAIQHESGILGPLLYGEVGDTLLIIFKNQASRPYNIYPHGITDVRPLYSRRLPKGVKHLKDFPILPGEIFKYKWTVTVEDGPTKSDPRCLTRYYSSFVNMERDLASGLIGPLLICYKESVDQRGNQIMSDKRNVILFSVFDENRSWYLTENIQRFLPNPAGVQLEDPEFQASNIMHSINGYVFDSLQLSVCLHEVAYWYILSIGAQTDFLSVFFSGYTFKHKMVYEDTLTLFPFSGETVFMSMENPGLWILGCHNSDFRNRGMTALLKVSSCDKNTGDYYEDSYEDISAYLLSKNNAIEPRSFSQNSRHPSQNPPVLKRHQREITRTTLQSDQEEIDYDDTISVEMKKEDFDIFDEDENQSPRSFQKKTRHYFIAAVERLWDYGMSSSPHVLRNRAQSGSVPQFKKVVFQEFTDGSFTQPLYRGELNEHLGLLGPYIRAEVEDNIMVTFRNQASRPYSFYSSLISYEEDQRQGAEPRKNFVKPNETKTYFWKVQHHMAPTKDEFDCKAWAYFSDVDLEKDVHSGLIGPLLVCHTNTLNPAHGRQVTVQEFALFFTIFDETKSWYFTENMERNCRAPCNIQMEDPTFKENYRFHAINGYIMDTLPGLVMAQDQRIRWYLLSMGSNENIHSIHFSGHVFTVRKKEEYKMALYNLYPGVFETVEMLPSKAGIWRVECLIGEHLHAGMSTLFLVYSNKCQTPLGMASGHIRDFQITASGQYGQWAPKLARLHYSGSINAWSTKEPFSWIKVDLLAPMIIHGIKTQGARQKFSSLYISQFIIMYSLDGKKWQTYRGNSTGTLMVFFGNVDSSGIKHNIFNPPIIARYIRLHPTHYSIRSTLRMELMGCDLNSCSMPLGMESKAISDAQITASSYFTNMFATWSPSKARLHLQGRSNAWRPQVNNPKEWLQVDFQKTMKVTGVTTQGVKSLLTSMYVKEFLISSSQDGHQWTLFFQNGKVKVFQGNQDSFTPVVNSLDPPLLTRYLRIHPQSWVHQIALRMEVLGCEAQDLY
ATRRYYLGAVELSWDYMQSDLGELPVDARFPPRVPKSFPFNTSVVYKKTLFVEFTDHLFNIAKPRPPWMGLLGPTIQAEVYDTVVITLKNMASHPVSLHAVGVSYWKASEGAEYDDQTSQREKEDDKVFPGGSHTYVWQVLKENGPMASDPLCLTYSYLSHVDLVKDLNSGLIGALLVCREGSLAKEKTQTLHKFILLFAVFDEGKSWHSETKNSLMQDRDAASARAWPKMHTVNGYVNRSLPGLIGCHRKSVYWHVIGMGTTPEVHSIFLEGHTFLVRNHRQASLEISPITFLTAQTLLMDLGQFLLFCHISSHQHDGMEAYVKVDSCPEEPQLRMKNNEEAEDYDDDLTDSEMDVVRFDDDNSPSFIQIRSVAKKHPKTWVHYIAAEEEDWDYAPLVLAPDDRSYKSQYLNNGPQRIGRKYKKVRFMAYTDETFKTREAIQHESGILGPLLYGEVGDTLLIIFKNQASRPYNIYPHGITDVRPLYSRRLPKGVKHLKDFPILPGEIFKYKWTVTVEDGPTKSDPRCLTRYYSSFVNMERDLASGLIGPLLICYKESVDQRGNQIMSDKRNVILFSVFDENRSWYLTENIQRFLPNPAGVQLEDPEFQASNIMHSINGYVFDSLQLSVCLHEVAYWYILSIGAQTDFLSVFFSGYTFKHKMVYEDTLTLFPFSGETVFMSMENPGLWILGCHNSDFRNRGMTALLKVSSCDKNTGDYYEDSYEDISAYLLSKNNAIEPRSFSQNSRHPSQNPPVLKRHQREITRTTLQSDQEEIDYDDTISVEMKKEDFDICDEDENQSPRSFQKKTRHYFIAAVERLWDYGMSSSPHVLRNRAQSGSVPQFKKVVFQEFTDGSFTQPLYRGELNEHLGLLGPYIRAEVEDNIMVTFRNQASRPYSFYSSLISYEEDQRQGAEPRKNFVKPNETKTYFWKVQHHMAPTKDEFDCKAWAYFSDVDLEKDVHSGLIGPLLVCHTNTLNPAHGRQVTVQEFALFFTIFDETKSWYFTENMERNCRAPCNIQMEDPTFKENYRFHAINGYIMDTLPGLVMAQDQRIRWYLLSMGSNENIHSIHFSGHVFTVRKKEEYKMALYNLYPGVFETVEMLPSKAGIWRVECLIGEHLHAGMSTLFLVYSNKCQTPLGMASGHIRDFQITASGQYGQWAPKLARLHYSGSINAWSTKEPFSWIKVDLLAPMIIHGIKTQGARQKFSSLYISQFIIMYSLDGKKWQTYRGNSTGTLMVFFGNVDSSGIKHNIFNPPIIARYIRLHPTHYSIRSTLRMELMGCDLNSCSMPLGMESKAISDAQITASSYFTNMFATWSPSKARLHLQGRSNAWRPQVNNPKEWLQVDFQKTMKVTGVTTQGVKSLLTSMYVKEFLISSSQDGHQWTLFFQNGKVKVFQGNQDSFTPVVNSLDPPLLTRYLRIHPQSWVHQIALRMEVLGCEAQDLY
内因性の全長FVIIIは、単鎖前駆体分子として合成される。分泌される前に、前駆体は、重鎖および軽鎖へと切断される。組換えBドメイン欠失FVIIIは、2つの異なる戦略により生成させることができる。Bドメインのない重鎖および軽鎖を2つの異なるポリペプチド鎖として個別に合成する(2本鎖戦略)か、またはBドメイン欠失FVIIIを、単鎖前駆体ポリペプチド鎖として合成し(単鎖戦略)、これを、全長FVIII前駆体と同じ形で重鎖および軽鎖へと切断する。
タンパク質のin vivo特性は、アルブミン結合側鎖を用いて向上させることができることが知られている。そのような側鎖またはアルブミン結合剤は、投与の前にタンパク質に結合させることができ、例えば、in vivoでタンパク質を安定させることができるか、タンパク質のin vivo半減期を向上もしくは延長させることができる。
(実施例1)
組換えBドメイン欠失型O-グリコシル化因子VIIIおよびその変異体、例えば因子VIII(Y1680F)または因子VIII(Y1680C)の生成
細胞系および培養工程
因子VIIIのcDNAを用いて、哺乳動物用の発現プラスミドを構築した。プラスミドは、Y1680F突然変異を含むBドメイン欠失因子VIII、全長ヒト因子VIIIのアミノ酸1〜740を含む因子VIII重鎖、および全長ヒト因子VIIIのアミノ酸1649〜2332を含む因子VIII軽鎖をコードする。重鎖配列および軽鎖配列は、全長ヒト因子VIIIのアミノ酸741〜750および1638〜1648による配列を含む21アミノ酸のリンカー(SFSQNSRHPSQNPPVLKRHQR:配列番号4)により連結した。配列番号4で定義されるリンカーを含むFVIII変異体は、本明細書で「N8」と呼ぶこともできる。このプラスミドによってコードされる因子VIIIアミノ酸配列は、配列番号1(wt)、配列番号2(Y1680F)、配列番号3(Y1680C)に示す通りである。
細胞培養培地から、Bドメイン欠失因子VIII(Y1680F)を単離するために、Capto MMCカラムにおける濃縮ステップ、ELISA(immunoabsorbent)およびクロマトグラフィーによるステップ、アニオン交換クロマトグラフィーステップ、ならびに最後のゲル濾過ステップを含めた、4ステップの精製手順を用いた。典型的には、以下の手順を使用した:11リットルの滅菌濾過培地を、緩衝液A:20mMのイミダゾール、10mMのCaCl2、50mMのNaCl、0.02%のTween 80、pH=7.5中で平衡化したCapto MMC(GE Healthcare、Sweden)によるカラム(1.6×12cm)へと、流速15ml/分で送入した。75mlの緩衝液Aでカラムを洗浄した後、1.5MのNaClを含有する75mlの緩衝液Aで洗浄した。20mMのイミダゾール、10mMのCaCl2、0.02%のTween 80、2.5MのNaCl、8Mのエチレングリコール、pH=7.5により、流速1ml/分でタンパク質を溶出させた。8mlの画分を回収し、因子VIII活性(FVIII:C)について比色アッセイでアッセイした(実施例3参照)。因子VIIIを含有する画分をプールし、通常約50mlのプール容量を得た。
組換えO-グリコシル化因子VIIIのペグ化の手法
以下の手法を用いて、実施例1で得られる組換え因子VIII分子をポリエチレングリコール(PEG)とコンジュゲートさせる。
水(pH7.3)中のイミダゾール(20mM)、塩化カルシウム(10mM)、ツイーン80(0.02%)、塩化ナトリウム(500mM)およびグリセロール(1M)からなる緩衝液中のBDD-FVIII Y1680F(1.18mg、0.85mg/ml)を解凍した。
Peg-30K-マレイミドによるY1680Cのペグ化(参照US2006/0115876 A1)
試薬:
1) BDD-FVIII N8-Y1680C(MW 178,000)、1200μl、濃度80μg/ml、96μg、緩衝液20mMイミダゾール、+10mM CaCl2、+0.02%ツイーン80、+1M NaCl、1M gグリセロール、pH7.3に0.54nmol
2)トリスカルボキシエチルホスフィン(TCEP、MW 287):700等量; 0.0315μmモル;9μg。1mgのTCEPを、1mlの緩衝液20mMイミダゾール、10mM CaCl2、0.02%ツイーン80、1M Gグリセロール、pH7.3、1M NaClに溶解させた。この溶液の109μlを用いた。
3) 30kDa PEG-マレイミド(NOF Corp.からのSunbright Me-300Ma、MW 29300)、10等量、180μg。4.8mgの30kDa PEG-マレイミドを、2.4mlの緩衝液20mMイミダゾール、10mM CaCl2、0.02%ツイーン80、1M Gグリセロール、pH7.3、1M NaClに溶解させた。この溶液の90μlを用いた。
緩衝液A: 20mMイミダゾール、10mM CaCl2、0.02%ツイーン80、1Mグリセロール、pH7.3
緩衝液B: 20mMイミダゾール、10mM CaCl2、0.02%ツイーン80、1Mグリセロール、pH7.3 25mM NaCl
緩衝液C:20mMイミダゾール、10mM CaCl2、0.02%ツイーン80、1Mグリセロール、pH7.3 50mM NaCl
緩衝液D:20mMイミダゾール、10mM CaCl2、0.02%ツイーン80、1Mグリセロール、pH7.3 200mM NaCl
緩衝液E:20mMイミダゾール、10mM CaCl2、0.02%ツイーン80、1Mグリセロール、pH7.3 1M NaCl
試料ローディング:希釈試料の4×11mlを加えた。各々について、2000gで2分間遠心回転させる。
溶出緩衝液B(25mM NaCl): 10mlの緩衝液Bを加え、2000gで2分間遠心回転させる。
溶出緩衝液C(50mM NaCl): 10mlの緩衝液Cを加え、2000gで2分間遠心回転させる。
溶出緩衝液D(200mM NaCl): 10mlの緩衝液Dを加え、2000gで2分間遠心回転させる。
溶出緩衝液E(1M NaCl): 2×750μlの緩衝液Eを加え、2000gで1分間遠心回転させる。(2回繰り返した)
発色性アッセイで測定されるFVIII:C
rFVIII化合物のFVIII活性(FVIII:C)は、以下の通りにCoatest SP試薬(Chromogenix)を用いて発色性FVIIIアッセイで評価された: rFVIII試料およびFVIII標準(例えばNIBSCからの第7国際FVIII標準に対して較正された、精製野生型rFVIII)を、Coatestアッセイ緩衝液(50mMトリス、150mM NaCl、1% BSA、pH7.3、保存剤含有)に希釈した。試料、標準および緩衝液陰性対照の50μlを、2反復で96穴マイクロタイタープレート(Nunc)に加えた。Coatest SPキットからの因子IXa/因子X試薬、リン脂質試薬およびCaCl2を5:1:3(vol:vol:vol)で混合し、これの75μlをウェルに加えた。室温で15分間のインキュベーションの後、50μlの因子Xa基質S-2765/トロンビン阻害剤I-2581ミックスを加え、反応体を室温で10分間インキュベートした後、25μlの1Mクエン酸、pH3を加えた。Spectramaxマイクロタイタープレートリーダー(Molecular Devices)で415nmの吸光度を測定し、620nmの吸光度を参照波長として用いた。陰性対照の値を全ての試料から引き、プロットされた吸光度値対FVIII濃度の線形回帰によって較正曲線を作成した。HPLCで測定されたタンパク質濃度で試料の活性を割ることによって、比活性を計算した。軽鎖に対応するクロマトグラム中のピーク下面積を積算し、濃度がアミノ酸分析で測定された野生型未改変rFVIIIの平行分析で同じピークの面積と比較することによって、試料濃度を決定した。O-グリコペグ化rFVIII化合物については比FVIII:C活性が維持されたことを、Table 1(表2)のデータは実証する。
1段階クロットアッセイで測定されるFVIII:C
rFVIII化合物のFVIII:Cは、以下の通りに1段階FVIIIクロットアッセイでさらに評価した: rFVIII試料およびFVIII標準(例えばNIBSCからの第7国際FVIII標準に対して較正された、精製野生型rFVIII)を、HBS/BSA緩衝液(1%BSAを含む20mM hepes、150mM NaCl、pH7.4)で約10U/mlに希釈し、その後VWFを含んでいるFVIII欠損血漿(Dade Behring)で10倍に希釈した。試料は、その後HBS/BSA緩衝液で希釈した。APTTクロット時間は、単一因子プログラムを用いるACL300RまたはACL5000機器(Instrumentation Laboratory)で測定した。VWFを含むFVIII欠損血漿(Dade Behring)をアッセイ血漿として用い、およびSynthASil(HemosIL(商標)、Instrumentation Laboratory)をaPTT試薬として用いた。クロット機器では、希釈された試料または標準を、FVIII欠損血漿、aPTT試薬と37℃で混合する。塩化カルシウムを加え、クロット形成までの時間を濁度で測定する。試料中のFVIII:Cは、FVIII標準の希釈溶液のクロット形成時間の標準曲線に基づいて計算される。Table 1(表2)のデータは、凝固と発色活性との間の比を示す。
FVIII欠損およびVWF欠損マウスでのrFVIIIの薬物動態
rfviii変異体の薬物動態は、FVIII欠損マウス(c57bl/6バックグラウンドを有するFVIIIエクソン16ノックアウト(KO)マウス、Taconic m&bで育成)で、またはvWF欠損マウス(c57bl/6バックグラウンドを有するvWFエクソン4 + 5 KOマウス、Charles River、Germanyで育成)で評価された。vWF-KOマウスは13%の正常なFVIII:Cを有していたが、FVIII-KOマウスは検出可能なFVIII:Cを有していなかった。25グラムの概算重量を有する16〜28週齢の雌雄(約1:1)混合群を用いた。マウスは、尾静脈からrFVIII(280iu/kg)の単一のi.v.注射を受けた。コーティングされていないガラス毛管を用いて、投与から最高64時間後までの時点で眼窩叢から採血した。3つの試料を各マウスからとり、2つから4つの試料を各時点で収集した。クエン酸ナトリウムで血液を直ちに安定させ、4容のFVIII coatest sp緩衝液(実施例4を参照)で希釈した後、4000×gで5分間の遠心にかけた。希釈血液から得られた血漿をドライアイスの上で冷凍し、-80℃で保存した。FVIII:Cは、実施例4に記載の発色性アッセイで測定した。winnonlinプロバージョン4.1ソフトウェアを用いて、ノンコンパートメント法(NCA)によって薬物動態分析を実行した。Table 2(表3)は、薬物動態パラメータの推定値を示す:半減期(t1/2)、クリアランス(cl)および平均滞留時間(MRT)。データは、ペグ化の結果クリアランスが減少し、半減期および平均滞留時間が増加したことを示す。
血友病AマウスのFeCl3によって誘導された損傷モデルにおけるvWF結合能力が欠如しているFVIII変異体の止血効果
マウスをケタミン(Ketaminol(登録商標) Vet.(Intervet);100mg/kg)、キシラジン(Narcoxyl(登録商標) Vet.注射溶液(Intervet);5.6mg/kg)およびアトロピン(硫酸アトロピン(Phoenix Pharma);0.3mg/kg)の混合物で麻酔にかけ、体温を維持するために加熱パッド(37℃)の上に置いた。その後頸動脈を露出させ、0.5PSB Nanoprobeを動脈周囲に置いた。FeCl3損傷の誘導の5分前に、試験化合物を側部尾静脈に注射した。10% FeCl3溶液に短時間浸した濾紙(2×5mm)をプローブに隣接した動脈の周囲に置き、3分後に除去することによってFeCl3損傷を誘導した。FeCl3飽和濾紙の除去後、動脈を0.9% NaClで3回洗浄し、最後に、流動プローブ中の空気を排気して血流の最適化された測定を確保するためにSurgilube(音響カプラ)を適用した。FeCl3の除去後、最初の血流があまりに低い(0.4ml/分未満)場合、または流動が測定不能の場合は、マウスを除外した。FeCl3飽和濾紙を除去してから25分間、AD instrumentsからのソフトウェアChart5(バージョン5.5.5.20)を用いて血流(ml/分)を記録した。
特記しない限りデータは平均±SEMで提示し、GraphPad Prismバージョン5(La Jolla、CA、USA)を用いて分析した。2つの化合物の効果は、一元配置ANOVAを用いて、続いて多重比較のためのチューキーの検定を用いて比較した。P<0.05は、統計的に有意であるとみなした。
FeCl3損傷の後、正常に機能している凝固系を有するマウス(C57BL/6NTac)の全ての血管は3.3分以内に閉塞し、閉塞までの平均時間は2.4±0.2分であった(n=6;図1)。FVIIIノックアウトマウスのいずれも、25分の観察時間中に閉塞しなかった(n=6)。NNC 0129-0000-9105の用量依存性は、閉塞までの時間を短縮した(NNC 0129-0000-9105は、欠失型Bドメイン中のO-連結グリカン上のF8-500 Y1680F 40kDa-ペグ化(PEGyleret)に対応する)。閉塞までの時間は、ビヒクル処置動物と比較して5または10IU/kgのいずれかによる処置の後、有意により短かった。Advate(登録商標)の類似した効果が観察され、Advate(登録商標)とNNC 0129-0000-9105との間で効果に統計的差はなかった(図1)。
血友病Aマウスの尾処理出血モデルにおけるvWF結合能力が欠如しているFVIII変異体の止血効果
マウスにペントバルビタール(100mg/kg、i.p.;Nomeco、Roskilde、Denmark)で麻酔をかけ、体温を維持するために加熱パッドの上に置いた。鋭いメスで尾部の先端の4mmを切断することによって、出血を誘導した。切断の10分前に、14mLの生理食塩水(37℃)を含む試験管に尾を入れ、切断の5分前に、尾静脈注射によって試験化合物を5mL/kgの量で投与した(20または280U/kg)。血友病Aマウスおよび正常な凝固をもつ動物を、同じ量のビヒクルで処置した。動物を安楽死させた後に、30分間にわたって血液を収集した。血液減少は、ヘモグロビンの量を定量化することによって測定した。したがって、4000×で5分間の遠心によって赤血球を単離した。上清を廃棄し、ヘモグロビン試薬(ABX Lysebio;HORIBA、ABX、Roedover、Denmark)で細胞を溶解した。4000×gで5分間の遠心によって細胞片を除去した。試料を550nmで読みとり、ヘモグロビン総量を標準曲線から決定した(ヘモグロビン標準;J.T. Baker 3074;Bie & Berntsen、Roedover、Denmark)。
特記しない限りデータは平均±SEMで提示し、GraphPad Prismバージョン5(La Jolla、CA、USA)を用いて分析した。2つの化合物の効果は、一元配置ANOVAを用いて、続いて多重比較のためのチューキーの検定を用いて比較した。P<0.05は、統計的に有意であるとみなされた。
FVIIIノックアウトマウスの尾部出血モデルにおけるAdvate(登録商標)の用量反応相関は、前に完全に調査されている(Documentum ObjectID: 0900c76e80e3e7a5)。200IU/kgのAdvate(登録商標)は血液減少および出血時間を正常化し、血液減少および出血時間のED50値はそれぞれ39および28IU/kgであった。NNC 0129-0000-9105の止血効果を試験するために、20および280IU/kgの効果をAdvate(登録商標)の同じ用量と比較した。NNC0129-0000-9105(20および280lU/kg)は、ビヒクル処置動物と比較して、血液減少および出血時間を有意に低減した(図2)。Advate(登録商標)とNNC 0129-0000-9105との間でその効果の差は観察されなかった。
vWFに結合しない血小板結合特性によるFVIII変異体の生成。
抗GPIIa/IIIB抗体の重鎖および軽鎖の可変領域、AP3を、SMART(商標)RACE cDNA増幅キット(Clontech、Ca)を用いてAP3抗体を発現するハイブリドーマ細胞から単離されたRNAから増幅した。2つのAP3鎖の可変領域の増幅のために用いたプライマーは、以下の通りであった:
重鎖:
汎用プライマーミックスA(Clontech、CA):
長い(0.4μM):
5'-ctaatacgactcactatagggcAAGCAGTGGTATCACGCAGAGT-3'(配列番号5)
短い(2μM):
5'-ctaatacgactcactatagggc-3'(配列番号6)
プライマー69(10μM)
5'-gctctagactaacactcattcctgttgaagctcttg-3'(配列番号7)
軽鎖
汎用プライマーミックスA(Clontech:
長い(0.4μM):
5'-ctaatacgactcactatagggcAAGCAGTGGTATCACGCAGAGT-3'(配列番号8)
短い(2μM):
5'-ctaatacgactcactatagggc-3'(配列番号9)
プライマー312(10μM)
5'-gtctaccacaacacacgtgac-3'(配列番号10)
DIVMTQAAPSVPVTPGESVSISCRSSRSLLHSNGNTYLCWFLQRPGQSPQLLIYRMSNLASGVPDRFSGSGSGTAFTLRISRVEAEDVGVYYCMQHLEYPFTFGSGTKLEIKRGGGGSGGGGSGGGGSQVQLQQSGAELVRPGTSVKISCKASGYTFTNYWLGWVKQRPGHGLEWIGDIYPGGGYNKYNENFKGKATLTADTSSSTAYMQLSSLTSEDSAVYFCAREYGNYDYAMDSWGQGTSVTVSSDYKDDDDK*
QVQLQQSGAELVRPGTSVKISCKASGYTFTNYWLGWVKQRPGHGLEWIGDIYPGGGYNKYNENFKGKATLTADTSSSTAYMQLSSLTSEDSAVYFCAREYGNYDYAMDSWGQGTSVTVSSGGGGSGGGGSGGGGSDIVMTQAAPSVPVTPGESVSISCRSSRSLLHSNGNTYLCWFLQRPGQSPQLLIYRMSNLASGVPDRFSGSGSGTAFTLRISRVEAEDVGVYYCMQHLEYPFTFGSGTKLEIKRDYKDDDDK*
3-(2,5-ジオキソ-2,5-ジヒドロピロール-1-イル)プロピオン酸2,5-ジオキソピロリジニル-1-イルエステル
MS: m/z=289、[M+Na]+のために必要とされる: 289
1H-NMR (CDCl3) δ 2.82 (m, 4H); 3.02 (t, 2H); 3.94 (t, 2H)、6.73 (s, 2H)。
N-((3-(ω-アミノ10kDaペギル)プロピオニルアミノ)アセチル)-O2-[5']シチジリル-ξ-ノイラミン酸(100mg、0.009mmol)を、テトラヒドロフラン(2ml)およびジクロロメタン(10ml)の混合物に溶解した。ジクロロメタン(3ml)中の3-(2,5-ジオキソ-2,5-ジヒドロピロール-1-イル)プロピオン酸2,5-ジオキソピロリジニル-1-イルエステル(50mg、0.18mmol)の溶液を加えた。エチルジイソプロピルアミン(0.005ml、0.028mmol)を加えた。反応混合液を室温で16時間撹拌した。ジクロロメタン(2ml)およびエチルジイソプロピルアミン(0.5ml)を加えた。アミノメチル化(Amionomethylated)ポリスチレン樹脂(例えばNovabiochemから市販されている、添加量0.85mmol/g、438mg、0.372mmol)を加えた。混合液を室温で1時間ゆっくりと撹拌した。樹脂を濾過によって除去した。25℃の浴温で溶媒を真空中で除去した。残基をジクロロメタン(4ml)に溶解した。エーテル(200ml)を加えた。形成された沈殿を加齢させるために、混合液を室温で2時間放置した。沈殿を濾過によって単離し、真空中で乾燥させ、標記化合物の38mgを与えた。DMSO-d6での1H-NMRスペクトルは、マレイミド基の存在を示した。
AP3 scFV C24S S248CのCys 248へのN-((3-(ω-(3-(2,5-ジオキソ-2,5-ジヒドロピロール-1-イル)プロピオニルアミノ)10kDaペギル)プロピオニルアミノ)アセチル)-O2-[5']シチジリル-ξ-ノイラミン酸の結合
pH7.35に調整した20mMイミダゾール、10mM CaCl2、150mM NaCl、0.02%ツイーン80および1Mのグリセロールからなる緩衝液(0.018ml)中の、重鎖のC末端にSFSQNSRHPSQNPPVLKRHQRの残留Bドメイン配列を有するBドメイン欠失FVIIIの溶液(1mg、5.64mmol)を、カットオフが10kDaのAmicon超遠心装置に入れた。pH8.0に調整した25mMトリスおよび150mM NaClの溶液(0.680ml)中のAP3 scFv C24S S248CのCys 248(1.29mg、34nmol)へのN-((3-(ω-(3-(2,5-ジオキソ-2,5-ジヒドロピロール-1-イル)プロピオニルアミノ)10kDaペギル)プロピオニルアミノ)-アセチル)-O2-[5']シチジリル-ξ-ノイラミン酸の給合生成物の溶液を加え、pH6.07に調整した20mMヒスチジン、10mM CaCl2、10%グリセロール、0.02%ツイーン80、500mM NaClからなる緩衝液(3ml)をその後加えた。混合液を、10℃で20分間の4000rpmの超遠心にかけた。FVIIIの残りの量は、0.800mlまたは1.25mg/mlであった。A.ウリファシエンス(A. Urifaciens)からのシアリダーゼの溶液(0.43mg/ml、302U/mg、0.0049ml、0.645U)およびST3-Gal-Iの溶液(2.5mg/ml、0.105mg、0.042ml)をその後加えた。反応混合液を32℃で1分間静かに振盪し、その後32℃で18時間放置した。それを精製までフリーザーで保存した。
N-((3-(ω-(9H-フルオレン-9-イルメトキシカルボニルアミノ)10kDaペギル)プロピオニルアミノ)アセチル)-O2-[5']シチジリル-ξ-ノイラミン酸(NNC 0129-0000-3219)
N-((3-(ω-アミノ10kDaペギル)プロピオニルアミノ)アセチル)-O2-[5']シチジリル-ξ-ノイラミン酸(NNC 0129-0000-3227)
4-ホルミル安息香酸2,5-ジオキソピロリジン-1-イルエステル
1H-NMR (CDCl3)。δ 2.95 (s, 4H); 8.04 (d, 2H)、8.32 (d, 2H); 10.15 (s, 1H)。
N-((3-(ω-アミノ10kDaペギル)プロピオニルアミノ)アセチル)-O2-[5']シチジリル-ξ-ノイラミン酸(42mg、0.004mmol)を、ジクロロメタン(2ml)に溶解した。エチルジイソプロピルアミン(0.002ml、0.012mmol)を溶液に加えた。ジクロロメタン(0.5ml)中の4-ホルミル安息香酸2,5-ジオキソピロリジン-1-イルエステルの溶液(19.32mg、0.078mmol)を加えた。反応混合液を室温で16時間撹拌した。溶媒を25℃の浴温で真空中で除去した。残渣を炭酸水素アンモニウムの25mM水溶液(15ml)に懸濁した。非可溶性物質を濾過によって除去した。それを5つの部分に分けた。その各々を、25mM炭酸水素アンモニウムの緩衝液を用いて、7ml/分の流動による直径26mmおよび長さ10cmのカラム上のG25を用いるサイズ除外クロマトグラフィーにかけた。所望の物質を含む全ての分画を合わせ、凍結乾燥した。DMSO-d6での1H-NMRスペクトルは、アルデヒド部分およびシチジリル部分の両方の存在を示した。得られた物質をフリーザーで保存した。
アブシキシマブ(Abxicimab)の1つのN末端へのN-((3-(ω-(4-ホルミルベンゾイルアミノ)10kDaペギル)プロピオニルアミノ)アセチル)-O2-[5']シチジリル-ξ-ノイラミン酸のコンジュゲーション
FVIIIのO-グリカンへのアブシキシマブのコンジュゲーション
pH7.35に調整した20mMヒスチジン、10mM CaCl2、150mM NaCl、0.02%ツイーン80および1Mグリセロールからなる緩衝液(2.5ml)を、pH7.35に調整した20mMイミダゾール、10mM CaCl2、150mM NaCl、0.02%ツイーン80および1Mグリセロールからなる緩衝液中の、重鎖のC末端にSFSQNSRHPSQNPPVLKRHQRの残留Bドメイン配列を有するBドメイン欠失FVIIIの溶液(5.7mg/ml、1mg、5.6nmol)に加えた。pH8.0に調整した25mMトリス、150mM NaClからなる緩衝液(0.323ml)中の、アブシキシマブ(Abxicimab)の1つのN末端とのN-((3-(ω-(4-ホルミルベンゾイルアミノ)10kDaペギル)プロピオニルアミノ)アセチル)-O2-[5']シチジリル-ξ-ノイラミン酸のコンジュゲートの溶液(2.3mg、39.5nmol)を加えた。溶液を10kDaのカットオフのAmicon超遠心装置に入れた。それを、10℃で20分間の4000rpmの超遠心にかけた。pH7.35に調整した20mMヒスチジン、10mM CaCl2、150mM NaCl、0.02%ツイーン80および1Mグリセロールからなる緩衝液(2ml)を加えた。溶液を10℃で30分間の4000rpmの超遠心にかけ、1.4mlの容量の溶液が残された。A.ウリファシエンス(A. Urifaciens)のシアリダーゼの溶液(0.4mg/ml、242U/mg、0.0066ml)およびST3Gal-Iの溶液(2.5mg/ml、0.042ml)をその後加えた。反応混合液を32℃で15分間静かに振盪した。その後、反応混合液を32℃で20.5時間静置させた。反応混合液を10kDaのカットオフのAmicon超遠心装置に入れた。それを、10℃で15分間の4000rpmの超遠心にかけた。0.300mlの残りの溶液は、pH7に調整した10mMヒスチジン、1.7mM CaCl2、0.01%ツイーン80、0.3M NaCl、8.8mMショ糖からなる緩衝液を溶離液として用い、0.5ml/分の流動の、10mm×300mmの床サイズのSuperose 6材料を用いるサイズ除外クロマトグラフィーにかけた。所望の生成物を含む分画をプールし、10kDaのカットオフのAmicon超遠心装置に入れた。pH6.07に調整した20mMヒスチジン、10mM CaCl2、10%グリセロール、0.02%ツイーン80、500mM NaClからなる緩衝液(2.5ml)を加えた。溶液を、10℃で15分間の4000rpmの超遠心にかけた。pH6.07に調整した20mMヒスチジン、10mM CaCl2、10%グリセロール、0.02%ツイーン80、500mM NaClからなる緩衝液(1.5ml)を加えた。溶液を、10℃で15分間の4000rpmの超遠心にかけた。0.220mlの残りの溶液をプラスチック反応器に入れた。pH6.07に調整した20mMヒスチジン、10mM CaCl2、10%グリセロール、0.02%ツイーン80、500mM NaClからなる緩衝液(0.173ml)中の市販のCMP NeuNAc(1.73mg、2800nmol)の溶液を加えた。反応混合液を32℃で1時間、300rpmで静かに振盪した。それを、pH7に調整した10mMヒスチジン、1.7mM CaCl2、0.01%ツイーン80、0.3M NaCl、8.8mMショ糖からなる緩衝液を溶離液として用い、0.5ml/分の流動の、10mm×300mmの床サイズのSuperose 6材料を用いるサイズ除外クロマトグラフィーにかけた。所望の生成物を含む分画をプールし、10kDaのカットオフのAmicon超遠心装置に入れた。プールを10℃で5分間の4000rpmの超遠心にかけ、O-グリカンでのFVIIIとのアブシキシマブのコンジュゲーション生成物の0.0358mgの0.275mlの溶液を与えた。定量化のために、13.15のモル吸光度をNanodrop(登録商標)分光光度計で用いた。SDS-PAGE分析は、O-グリカンでのFVIIIとのアブシキシマブのコンジュゲーション生成物のSDS-PAGEの予想に一致した。
Claims (14)
- 残基1670〜1684にわたる領域における1または複数のアミノ酸の点突然変異および/または欠失突然変異に起因する低減されたvWF結合能力を有し、少なくとも1つの側鎖基と共有結合でコンジュゲートしており、且つ、増加した循環半減期を有する、組換え因子VIII分子。
- 側鎖基が、親水性ポリマー、アルブミン結合剤、抗体またはその断片、およびアルブミンからなる群の1つまたは複数から選択される、請求項1に記載の分子。
- 因子VIII分子がBドメイン欠失型変異体である、請求項1または2のいずれか一項に記載の分子。
- 側鎖基が欠失型Bドメインと共有結合でコンジュゲートし、因子VIIIの活性化が、共有結合でコンジュゲートしている側鎖基の除去をもたらす、請求項3に記載の分子。
- 欠失型BドメインのO-連結オリゴ糖を通して親水性ポリマーと共有結合でコンジュゲートし、因子VIIIの活性化が、共有結合でコンジュゲートしている親水性ポリマーの除去をもたらす、請求項4に記載の分子。
- O-連結オリゴ糖がBドメインの欠失によって形成されるO-グリコシル化部位に結合する、請求項5に記載の分子。
- 残基1680に点突然変異を含む、請求項1〜6のいずれか一項に記載の分子。
- 残基1670〜1684にわたる領域で1つまたは複数のアミノ酸の欠失を含む、請求項1〜6のいずれか一項に記載の分子。
- 側鎖基がPEGである、請求項1〜8のいずれか一項に記載の分子。
- 低減されたvWF結合能力を有する因子VIII分子への側鎖基の結合を含む、請求項1〜9のいずれか一項に記載の分子の作製方法。
- 血友病疾患の治療のための、請求項1〜9のいずれか一項に記載の分子の治療的有効量を含む、医薬組成物。
- 医薬としての、請求項1〜9のいずれか一項に記載の分子を含む、医薬組成物。
- 血友病の治療のための医薬の製造のための、請求項1〜9のいずれか一項に記載の分子の使用。
- 請求項1〜9のいずれか一項に記載の分子を含む、医薬組成物。
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PCT/EP2011/051438 WO2011101242A1 (en) | 2010-02-16 | 2011-02-02 | Factor viii molecules with reduced vwf binding |
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Families Citing this family (47)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS6263355A (ja) * | 1985-09-13 | 1987-03-20 | Hitachi Ltd | バツテリバツクアツプramのチエツク方法 |
WO2008074032A1 (en) | 2006-12-15 | 2008-06-19 | Baxter International Inc. | Factor viia- (poly) sialic acid conjugate having prolonged in vivo half-life |
CN101778859B (zh) | 2007-06-12 | 2014-03-26 | 诺和诺德公司 | 改良的用于生产核苷酸糖的方法 |
PL2393828T3 (pl) | 2009-02-03 | 2017-06-30 | Amunix Operating Inc. | Wydłużone rekombinowane polipeptydy i zawierające je kompozycje |
NZ623810A (en) | 2009-07-27 | 2015-10-30 | Lipoxen Technologies Ltd | Glycopolysialylation of non-blood coagulation proteins |
US8809501B2 (en) | 2009-07-27 | 2014-08-19 | Baxter International Inc. | Nucleophilic catalysts for oxime linkage |
US8642737B2 (en) | 2010-07-26 | 2014-02-04 | Baxter International Inc. | Nucleophilic catalysts for oxime linkage |
AU2010277438B2 (en) | 2009-07-27 | 2015-08-20 | Baxalta GmbH | Glycopolysialylation of non-blood coagulation proteins |
ES2597954T3 (es) | 2009-07-27 | 2017-01-24 | Baxalta GmbH | Conjugados de proteína de la coagulación sanguínea |
EP2470559B1 (en) | 2009-08-24 | 2017-03-22 | Amunix Operating Inc. | Coagulation factor ix compositions and methods of making and using same |
US8765432B2 (en) | 2009-12-18 | 2014-07-01 | Oligasis, Llc | Targeted drug phosphorylcholine polymer conjugates |
GB201007356D0 (en) | 2010-04-30 | 2010-06-16 | Leverton Licence Holdings Ltd | Conjugated factor VIIa |
PT2654794T (pt) | 2010-12-22 | 2020-06-11 | Baxalta Inc | Materiais e métodos para conjugação de um derivado de ácido gordo solúvel em água a uma proteína |
MX357403B (es) * | 2012-01-12 | 2018-07-09 | Bioverativ Therapeutics Inc | Polipeptidos de factor viii quimericos y usos de los mismos. |
WO2013123457A1 (en) | 2012-02-15 | 2013-08-22 | Biogen Idec Ma Inc. | Recombinant factor viii proteins |
MY188897A (en) | 2012-02-15 | 2022-01-12 | Bioverativ Therapeutics Inc | Factor viii compositions and methods of making and using same |
EP2838566A2 (en) | 2012-04-16 | 2015-02-25 | Cantab Biopharmaceuticals Patents Limited | Optimised subcutaneous therapeutic agents |
PL2882450T3 (pl) * | 2012-07-11 | 2020-06-29 | Bioverativ Therapeutics Inc. | Kompleks czynnika viii z xten i białkiem czynnika von willebranda oraz jego zastosowania |
US20150259665A1 (en) | 2012-10-15 | 2015-09-17 | Novo Nordisk Health Care Ag | Factor vii conjugates |
EP2796145B1 (en) * | 2013-04-22 | 2017-11-01 | CSL Ltd. | A covalent complex of von willebrand factor and faktor viii linked by a disulphide bridge |
US10548953B2 (en) | 2013-08-14 | 2020-02-04 | Bioverativ Therapeutics Inc. | Factor VIII-XTEN fusions and uses thereof |
US10702608B2 (en) * | 2013-09-08 | 2020-07-07 | Kodiak Sciences Inc. | Factor VIII zwitterionic polymer conjugates |
MX2016004702A (es) | 2013-10-15 | 2016-07-22 | Novo Nordisk Healthcare Ag | Polipeptidos del factor vii de coagulacion. |
JP6749836B2 (ja) | 2014-01-10 | 2020-09-02 | バイオベラティブ セラピューティクス インコーポレイテッド | 第viii因子キメラタンパク質及びその使用 |
AR099340A1 (es) * | 2014-02-12 | 2016-07-13 | Novo Nordisk As | Conjugados del factor de coagulación ix |
AR101060A1 (es) * | 2014-02-12 | 2016-11-23 | Novo Nordisk As | Conjugados de fviii |
AR099328A1 (es) * | 2014-02-12 | 2016-07-13 | Novo Nordisk As | Conjugados de factor vii |
EP3122770A4 (en) * | 2014-03-23 | 2017-08-23 | Advantech Bioscience Farmacêutica Ltda. | Enhancement of recombinant protein expression with copper |
US9840553B2 (en) | 2014-06-28 | 2017-12-12 | Kodiak Sciences Inc. | Dual PDGF/VEGF antagonists |
KR102192494B1 (ko) | 2014-08-04 | 2020-12-18 | 시에스엘 리미티드 | 인자 viii 제형 |
KR20210013299A (ko) | 2014-10-17 | 2021-02-03 | 코디악 사이언시스 인코포레이티드 | 부티릴콜린에스테라제 양성이온성 중합체 컨쥬게이트 |
EP3042952A1 (en) * | 2015-01-07 | 2016-07-13 | CEVEC Pharmaceuticals GmbH | O-glycan sialylated recombinant glycoproteins and cell lines for producing the same |
SG11201706659WA (en) | 2015-03-06 | 2017-09-28 | Csl Behring Recombinant Facility Ag | Modified von willebrand factor having improved half-life |
UA126016C2 (uk) | 2015-08-03 | 2022-08-03 | Біовератів Терапеутікс Інк. | Злитий білок фактора іх |
KR20180104635A (ko) * | 2015-12-30 | 2018-09-21 | 코디악 사이언시스 인코포레이티드 | 항체 및 이의 접합체 |
EP3476860A4 (en) * | 2016-06-24 | 2020-01-22 | Mogam Institute for Biomedical Research | RECOMBINANT SINGLE CHAIN FVIII AND CHEMICAL CONJUGATE THEREOF |
CN109790529A (zh) | 2016-06-24 | 2019-05-21 | 财团法人牧岩生命科学研究所 | 包含FVIII和vWF因子的嵌合蛋白及其用途 |
EP3382014A1 (en) | 2017-03-29 | 2018-10-03 | CEVEC Pharmaceuticals GmbH | Recombinant glycoproteins with reduced antennary fucosylation |
JP7253505B2 (ja) * | 2017-06-23 | 2023-04-06 | 武田薬品工業株式会社 | 第viii因子亜種の精製 |
EP3441471A1 (en) | 2017-08-08 | 2019-02-13 | CEVEC Pharmaceuticals GmbH | Use of constitutively active variants of growth factor receptors as selection makers for the generation of stable producer cell lines |
AU2019227997A1 (en) | 2018-03-02 | 2020-09-24 | Kodiak Sciences Inc. | IL-6 antibodies and fusion constructs and conjugates thereof |
BR112020023168A2 (pt) * | 2018-05-18 | 2021-02-09 | Zhengzhou Gensciences Inc. | proteína de fusão de fviii aprimorada e uso da mesma |
CN112512555A (zh) | 2018-05-18 | 2021-03-16 | 比奥维拉迪维治疗股份有限公司 | 治疗血友病a的方法 |
KR20220029733A (ko) | 2019-07-04 | 2022-03-08 | 체에스엘 베링 렝나우 아게 | 응고 인자 viii의 시험관내 안정성을 증가시키기 위한 절단된 폰 빌레브란트 인자 (vwf) |
WO2021038296A2 (en) | 2019-08-27 | 2021-03-04 | Tonix Pharma Holdings Limited | Modified tff2 polypeptides |
EP4041312A4 (en) | 2019-10-10 | 2023-12-20 | Kodiak Sciences Inc. | METHOD FOR TREATING AN EYE DISORDER |
EP4058049A1 (en) | 2019-11-11 | 2022-09-21 | CSL Behring Lengnau AG | Polypeptides for inducing tolerance to factor viii |
Family Cites Families (35)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4356170A (en) | 1981-05-27 | 1982-10-26 | Canadian Patents & Development Ltd. | Immunogenic polysaccharide-protein conjugates |
AU2645588A (en) | 1987-12-04 | 1989-06-15 | Scripps Clinic And Research Foundation | The von willebrand factor binding domain of factor viii |
US5629384A (en) | 1994-05-17 | 1997-05-13 | Consiglio Nazionale Delle Ricerche | Polymers of N-acryloylmorpholine activated at one end and conjugates with bioactive materials and surfaces |
US5824784A (en) | 1994-10-12 | 1998-10-20 | Amgen Inc. | N-terminally chemically modified protein compositions and methods |
DE4437544A1 (de) * | 1994-10-20 | 1996-04-25 | Behringwerke Ag | Einsatz von vWF-enthaltenden Konzentraten als Kombinationstherapie bei Therapie mit Antithrombotika und Fibrinolytika |
US5932462A (en) | 1995-01-10 | 1999-08-03 | Shearwater Polymers, Inc. | Multiarmed, monofunctional, polymer for coupling to molecules and surfaces |
WO1999055377A2 (en) | 1998-04-28 | 1999-11-04 | Applied Research Systems Ars Holding N.V. | Polyol-ifn-beta conjugates |
US6759216B1 (en) * | 1998-11-06 | 2004-07-06 | Emory University | Glycosylated, low antigenicity low immunogenicity factor VIII |
US7157277B2 (en) * | 2001-11-28 | 2007-01-02 | Neose Technologies, Inc. | Factor VIII remodeling and glycoconjugation of Factor VIII |
CN102180944A (zh) | 2001-10-10 | 2011-09-14 | 诺和诺德公司 | 肽的重构和糖缀合 |
JP4758608B2 (ja) * | 2001-11-07 | 2011-08-31 | ネクター セラピューティックス | 分枝ポリマーおよびそれらの結合体 |
WO2003062290A1 (en) | 2002-01-16 | 2003-07-31 | Biocompatibles Uk Limited | Polymer conjugates |
EP1654290B1 (en) | 2003-08-12 | 2019-03-13 | Lipoxen Technologies Limited | Sialic acid derivatives for protein derivatisation and conjugation |
CN103214569B (zh) | 2004-11-12 | 2016-12-28 | 拜尔健康护理有限责任公司 | Fviii的位点定向修饰 |
WO2006090119A1 (en) | 2005-02-23 | 2006-08-31 | Lipoxen Technologies Limited | Activated sialic acid derivatives for protein derivatisation and conjugation |
ES2374935T3 (es) | 2005-03-24 | 2012-02-23 | Biogenerix Ag | Expresión de glicosiltransferasas eucariotas activas, solubles en organismos procariotas. |
US20080227691A1 (en) * | 2005-04-01 | 2008-09-18 | Novo Nordisk Health Care Ag | Blood Coagulation FVIII Analogues |
AU2006259080A1 (en) | 2005-06-15 | 2006-12-21 | Novo Nordisk Health Care Ag | Transglutaminase mediated conjugation of growth hormone |
US20070105755A1 (en) * | 2005-10-26 | 2007-05-10 | Neose Technologies, Inc. | One pot desialylation and glycopegylation of therapeutic peptides |
WO2007056191A2 (en) | 2005-11-03 | 2007-05-18 | Neose Technologies, Inc. | Nucleotide sugar purification using membranes |
EP1981977B1 (en) | 2006-01-31 | 2015-12-23 | National Research Council Of Canada | Production of polysialic acid containing glyconjugates using a self-priming polysialyltransferase |
US7683158B2 (en) | 2006-03-31 | 2010-03-23 | Baxter International Inc. | Pegylated factor VIII |
US7645860B2 (en) | 2006-03-31 | 2010-01-12 | Baxter Healthcare S.A. | Factor VIII polymer conjugates |
WO2008011633A2 (en) | 2006-07-21 | 2008-01-24 | Neose Technologies, Inc. | Glycosylation of peptides via o-linked glycosylation sequences |
WO2008074032A1 (en) | 2006-12-15 | 2008-06-19 | Baxter International Inc. | Factor viia- (poly) sialic acid conjugate having prolonged in vivo half-life |
EP2162535A4 (en) | 2007-06-04 | 2011-02-23 | Novo Nordisk As | O-linked glycosylation using N-acetylglucosamine transferases |
CN101730739B (zh) | 2007-06-15 | 2013-07-31 | 加拿大国家研究院 | 经工程改造的酶活性改善的聚唾液酸转移酶及其用途 |
CA2704234A1 (en) * | 2007-11-09 | 2009-05-14 | Baxter International Inc. | Modified recombinant factor viii and von willebrand factor and methods of use |
CA2711503A1 (en) | 2008-01-08 | 2009-07-16 | Biogenerix Ag | Glycoconjugation of polypeptides using oligosaccharyltransferases |
MX2010009154A (es) * | 2008-02-27 | 2010-09-09 | Novo Nordisk As | Moleculas conjugadas del factor viii. |
KR101507718B1 (ko) | 2008-06-24 | 2015-04-10 | 체에스엘 베링 게엠베하 | 연장된 생체내 반감기를 갖는 인자 viii, 폰 빌레브란트 인자 또는 이들의 복합체 |
ES2692172T3 (es) | 2008-10-17 | 2018-11-30 | Baxalta GmbH | Factores sanguíneos modificados que comprenden un bajo grado de polímero soluble en agua |
CN102333788A (zh) | 2009-02-19 | 2012-01-25 | 诺沃—诺迪斯克有限公司 | 因子viii的修饰 |
CN102482340B (zh) * | 2009-04-06 | 2015-05-13 | 诺沃—诺迪斯克有限公司 | 因子viii蛋白向血小板的靶向递送 |
JP2013519699A (ja) * | 2010-02-16 | 2013-05-30 | ノヴォ ノルディスク アー/エス | 因子viii融合タンパク質 |
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- 2011-02-02 WO PCT/EP2011/051438 patent/WO2011101242A1/en active Application Filing
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US20170020992A1 (en) | 2017-01-26 |
US9018166B2 (en) | 2015-04-28 |
WO2011101267A1 (en) | 2011-08-25 |
US20130040888A1 (en) | 2013-02-14 |
CN102884077B (zh) | 2016-06-08 |
CN102770449B (zh) | 2016-02-24 |
JP5933457B2 (ja) | 2016-06-08 |
JP2013519636A (ja) | 2013-05-30 |
US20150376262A1 (en) | 2015-12-31 |
EP2536755A1 (en) | 2012-12-26 |
CN102884077A (zh) | 2013-01-16 |
WO2011101242A1 (en) | 2011-08-25 |
CN102770449A (zh) | 2012-11-07 |
JP2013519697A (ja) | 2013-05-30 |
EP2536753B1 (en) | 2017-12-20 |
US20120322738A1 (en) | 2012-12-20 |
CN105524164A (zh) | 2016-04-27 |
EP2536753A1 (en) | 2012-12-26 |
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