JP5759102B2 - Vii因子・組織因子共有結合複合体 - Google Patents
Vii因子・組織因子共有結合複合体 Download PDFInfo
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- JP5759102B2 JP5759102B2 JP2009503550A JP2009503550A JP5759102B2 JP 5759102 B2 JP5759102 B2 JP 5759102B2 JP 2009503550 A JP2009503550 A JP 2009503550A JP 2009503550 A JP2009503550 A JP 2009503550A JP 5759102 B2 JP5759102 B2 JP 5759102B2
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Description
本発明は、VII因子ポリペプチドと組織因子ポリペプチドの新規な共有結合複合体、特に機能的に活性で、対応する遊離のVII因子ポリペプチドと比較してX因子に対して亢進されたタンパク分解活性を有する該複合体に関する。かかる複合体は出血性疾患の治療に特に有用であると考えられる。
高められた生物学的活性を有するVIIa因子ポリペプチドは、組換え型VIIa因子の最適以下の作用強度のため一般的な組換え型VIIa因子療法には部分的もしくは完全に不応性である止血不能の出血の治療のための望ましい分子である。作用強度を高めるための現在のアプローチ法は、組織因子とは無関係に、活性化血小板に対する直接のX因子活性化であると思われる組換え型VIIa因子の作用機序を反映させている。
本発明は、VII因子ポリペプチドと組織因子ポリペプチドの新規な共有結合複合体に関する。該複合体は、対応する遊離のVII因子ポリペプチドに対して、巨大分子基質X因子に対する亢進されたタンパク分解活性を示す。
本発明はとりわけVII因子ポリペプチドと組織因子ポリペプチドの共有結合複合体であって、VII因子ポリペプチドと組織因子ポリペプチドが、(i)VII因子ポリペプチドのアミノ酸と(ii)組織因子ポリペプチドのアミノ酸のセットのアミノ酸側鎖間の一又は複数の指定された結合によって共有結合されている複合体に関する。
ある重要な実施態様では、VII因子ポリペプチドは機能的に活性である。
ここで使用される「野生型ヒトFVIIa」は米国特許第4784950号に開示されたアミノ酸配列を有するポリペプチドである。
304Argから329Cysまでのアミノ酸配列における置換、付加又は欠失;及び153Ileから223Argまでのアミノ酸配列における置換、付加又は欠失が含まれる。
Ser1−Gly−Thr−Thr−Asn−Thr−Val−Ala−Ala−Tyr10−Asn−Leu−Thr−Trp−Lys−Ser−Thr−Asn−Phe−Lys20−Thr−Ile−Leu−Glu−Trp−Glu−Pro−Lys−Pro−Val30−Asn−Gln−Val−Tyr−Thr−Val−Gln−Ile−Ser−Thr40−Lys−Ser−Gly−Asp−Trp−Lys−Ser−Lys−Cys−Phe50−Tyr−Thr−Thr−Asp−Thr−Glu−Cys−Asp−Leu−Thr60−Asp−Glu−Ile−Val−Lys−Asp−Val−Lys−Gln−Thr70−Tyr−Leu−Ala−Arg−Val−Phe−Ser−Tyr−Pro−Ala80−Gly−Asn−Val−Glu−Ser−Thr−Gly−Ser−Ala−Gly90−Glu−Pro−Leu−Tyr−Glu−Asn−Ser−Pro−Glu−Phe100−Thr−Pro−Tyr−Leu−Glu−Thr−Asn−Leu−Gly−Gln110−Pro−Thr−Ile−Gln−Ser−Phe−Glu−Gln−Val−Gly120−Thr−Lys−Val−Asn−Val−Thr−Val−Glu−Asp−Glu130−Arg−Thr−Leu−Val−Arg−Arg−Asn−Asn−Thr−Phe140−Leu−Ser−Leu−Arg−Asp−Val−Phe−Gly−Lys−Asp150−Leu−Ile−Tyr−Thr−Leu−Tyr−Tyr−Trp−Lys−Ser160−Ser−Ser−Ser−Gly−Lys−Lys−Thr−Ala−Lys−Thr170−Asn−Thr−Asn−Glu−Phe−Leu−Ile−Asp−Val−Asp180−Lys−Gly−Glu−Asn−Tyr−Cys−Phe−Ser−Val−Gln190−Ala−Val−Ile−Pro−Ser−Arg−Thr−Val−Asn−Arg200−Lys−Ser−Thr−Asp−Ser−Pro−Val−Glu−Cys−Met210−Gly−Gln−Glu−Lys−Gly−Glu−Phe−Arg−Glu−Ile220−Phe−Tyr−Ile−Ile−Gly−Ala−Val−Val−Phe−Val230−Val−Ile−Ile−Leu−Val−Ile−Ile−Leu−Ala−Ile240−Ser−Leu−His−Lys−Cys−Arg−Lys−Ala−Gly−Val250−Gly−Gln−Ser−Trp−Lys−Glu−Asn−Ser−Pro−Leu260−Asn−Val−Ser263(配列番号2)。
等々である。
好ましくは、複合体種の少なくとも95%、又は更には少なくとも98%、又は実質的に全部が複合体種の集団全体のセットの各々内の各アミノ酸の位置及びタイプに対して同一である。
よって、本発明は、VII因子ポリペプチドと組織因子ポリペプチドが、(i)VII因子ポリペプチドのアミノ酸と(ii)組織因子ポリペプチドのアミノ酸のセットのアミノ酸側鎖間の一又は複数の直接結合によって共有結合されている上に定義された複合体を提供する。
一変形例では、セットの各々は、例えばヘテロ二官能性試薬へのカップリングの際に使用される1個のシステインアミノ酸を含んでいる。
他の変形例では、セットの各々は2個のシステインアミノ酸を含んでいる。この変形例では、ジスルフィド結合が樹立される。
ある重要な実施態様では、可溶型組織因子ポリペプチド中の位置Thr17、Lys20、Ile22、Ser42、Gly43、Asp44、Trp45、Lys46、Ser47、Phe50、Cys57、Asp58、Asp61、Gly109、Gln110、Leu133、Ser205、Pro206及びVal207、特に可溶型組織因子ポリペプチド中のTrp45、Asp58、Gly109、Pro206及びVal207のアミノ酸の少なくとも一つが、指定された結合、例えば特定結合、特に直接結合を生じる反応性側鎖を有するアミノ酸によって置換されている。好ましい変形例では、該少なくとも一つのアミノ酸はシステインアミノ酸である。
FVII−Ile69 − sTF−Thr17、
FVII−Ile69 − sTF−Lys20、
FVII−Arg79 − sTF−Ile22、
FVII−Val92 − sTF−Ser47、
FVII−Val92 − sTF−Phe50、
FVII−Gly78 − sTF−Cys57、
FVII−Glu77 − sTF−Asp58、
FVII−Gly78 − sTF−Asp58、
FVII−Arg79 − sTF−Asp58、
FVII−Arg277 − sTF−Ser42、
FVII−Val276 − sTF−Gly43、
FVII−Arg277 − sTF−Gly43、
FVII−Phe275 − sTF−Asp44、
FVII−Val276 − sTF−Asp44、
FVII−Arg277 − sTF−Asp44、
FVII−Phe275 − sTF−Trp45、
FVII−Val276 − sTF−Trp45、
FVII−Arg134 − sTF−Trp45、
FVII−Phe275 − sTF−Lys46、
FVII−Gln64 − sTF−Gly109、
FVII−Gln64 − sTF−Gln110、
FVII−Leu65 − sTF−Gln110、
FVII−Phe71 − sTF−Leu133、
FVII−Ser43 − sTF−Ser205、
FVII−Leu39 − sTF−Pro206、
FVII−Phe40 − sTF−Pro206、
FVII−Ile42 − sTF−Pro206、
FVII−Ser43 − sTF−Pro206、
FVII−Tyr44 − sTF−Pro206、
FVII−Leu39 − sTF−Val207、
FVII−Phe40 − sTF−Val207、及び
FVII−Ser43 − sTF−Val207
の少なくとも一つ、
特にアミノ酸セット
FVII−Phe40 − sTF−Val207、
FVII−Ser43 − sTF−Pro206、
FVII−Gln64 − sTF−Gly109、
FVII−Glu77 − sTF−Asp58、及び
FVII−Phe275 − sTF−Trp45
の少なくとも一つのアミノ酸の一方又は双方が、指定された結合、例えば特定結合、特に直接結合を生じる反応性側鎖を有するアミノ酸によって置換されている。
他の変形例では、少なくとも一つの直接結合は、特に、photo−Ile、photo−Leu及びphoto−Metからなる群から選択される光反応性アミノ酸を介して形成される。
ヘテロ二官能性試薬を使用する組織因子とVII因子の架橋
一つの興味深い変形例では、複合体の調製方法は、可溶型組織因子のシステイン変異体を生産し、ついで可溶型組織因子中のシステインを、官能基の一つがシステイン反応性であるヘテロ二官能性試薬で標識し、最後に試薬の第二官能基によってVIIa因子に架橋させることを含む。大腸菌中での組織因子のシステイン変異体のクローニング及び発現並びにシステイン特異的試薬での標識の方法は過去に記載されている(Stone等(1995) Biochem. J., 310, 605-614;Freskgard等(1996) Protein Sci., 5, 1521-1540;Owenius等(1999) Biophys. J., 77, 2237-2250;Osterlund等(2001) Biochemistry, 40, 9324-9328)。一つの特異的システイン及び一つの光活性化可能な官能基を含むヘテロ二官能性試薬を使用するタンパク質の光架橋はZhang等(1995) Biochem. Biophys. Res. Commun., 217, 1177-1184に記載されている。
一つの特に興味深い変形例では、複合体の調製方法は、VII因子ポリペプチドと組織因子ポリペプチドの同時発現を含み、これにより、2つのポリペプチド間に共有結合を直ぐに樹立させることができる。
材料 − D−Phe−Phe−Arg−クロロメチルケトンをBachemから購入した。発色Z−D−Arg−Gly−Arg−p−ニトロアニリド(S−2765)、及びH−D−Ile−Pro−Arg−p−ニトロアニリド(S−2288)基質をChromogenix(Sweden)から得た。ヒト血漿由来X因子(hFX)、Xa因子(hFXa)、及びIXa因子(hFIXa)をEnzyme・Research・Laboratories社(South Bend, IN)から得た。ヒト全脳マラソン既成cDNAライブラリーをClontech(Mountain View, CA)から得た。大腸菌中に発現された可溶型組織因子1−219(sTF(1−219))は公開された手順(Freskgard等(1996) Protein Sci., 5, 1531-1540)に従って調製した。組換え型VIIa因子の発現と精製を、過去に記載されているようにして(Thim等(1988) Biochemistry, 27, 7785-7793;Persson等(1996) FEBS Lett., 385, 241-243)実施した。他の全ての化学物質は分析用等級又はそれ以上のものであった。
sTF(1−219)V207CをコードするプラスミドpHOJ357は、製造者(Stratagene, La Jolla, CA)の指示書に従って、プライマーoHOJ144−f及びoHOJ144−rと鋳型としてpHOJ356を用いるQuickChange(登録商標)部位特異的突然変異誘発によって構築した。全てのクローン化配列の正確な同一性はDNA配列決定によって実証した。
比=((A405nmVIIa因子−sTF複合体)/[VIIa因子−sTF複合体])/((A405nmVIIa因子野生型)/[VIIa因子野生型])
結果は表2に与える。
Ala1-Asn-Ala-Phe-Leu-GLA-GLA-Leu-Arg-Pro10-Gly-Ser-Leu-GLA-Arg-GLA-Cys-Lys-GLA-GLA20-Gln-Cys-Ser-Phe-GLA-GLA-Ala-Arg-GLA-Ile30-Phe-Lys-Asp-Ala-GLA-Arg-Thr-Lys-Leu-Phe40-Trp-Ile-Ser-Tyr-Ser-Asp-Gly-Asp-Gln-Cys50-Ala-Ser-Ser-Pro-Cys-Gln-Asn-Gly-Gly-Ser60-Cys-Lys-Asp-Gln-Leu-Gln-Ser-Tyr-Ile-Cys70-Phe-Cys-Leu-Pro-Ala-Phe-Glu-Gly-Arg-Asn80-Cys-Glu-Thr-His-Lys-Asp-Asp-Gln-Leu-Ile90-Cys-Val-Asn-Glu-Asn-Gly-Gly-Cys-Glu-Gln100-Tyr-Cys-Ser-Asp-His-Thr-Gly-Thr-Lys-Arg110-Ser-Cys-Arg-Cys-His-Glu-Gly-Tyr-Ser-Leu120-Leu-Ala-Asp-Gly-Val-Ser-Cys-Thr-Pro-Thr130-Val-Glu-Tyr-Pro-Cys-Gly-Lys-Ile-Pro-Ile140-Leu-Glu-Lys-Arg-Asn-Ala-Ser-Lys-Pro-Gln150-Gly-Arg-Ile-Val-Gly-Gly-Lys-Val-Cys-Pro160-Lys-Gly-Glu-Cys-Pro-Trp-Gln-Val-Leu-Leu170-Leu-Val-Asn-Gly-Ala-Gln-Leu-Cys-Gly-Gly180-Thr-Leu-Ile-Asn-Thr-Ile-Trp-Val-Val-Ser190-Ala-Ala-His-Cys-Phe-Asp-Lys-Ile-Lys-Asn200-Trp-Arg-Asn-Leu-Ile-Ala-Val-Leu-Gly-Glu210-His-Asp-Leu-Ser-Glu-His-Asp-Gly-Asp-Glu220-Gln-Ser-Arg-Arg-Val-Ala-Gln-Val-Ile-Ile230-Pro-Ser-Thr-Tyr-Val-Pro-Gly-Thr-Thr-Asn240-His-Asp-Ile-Ala-Leu-Leu-Arg-Leu-His-Gln250-Pro-Val-Val-Leu-Thr-Asp-His-Val-Val-Pro260-Leu-Cys-Leu-Pro-Glu-Arg-Thr-Phe-Ser-Glu270-Arg-Thr-Leu-Ala-Phe-Val-Arg-Phe-Ser-Leu280-Val-Ser-Gly-Trp-Gly-Gln-Leu-Leu-Asp-Arg290-Gly-Ala-Thr-Ala-Leu-Glu-Leu-Met-Val-Leu300-Asn-Val-Pro-Arg-Leu-Met-Thr-Gln-Asp-Cys310-Leu-Gln-Gln-Ser-Arg-Lys-Val-Gly-Asp-Ser320-Pro-Asn-Ile-Thr-Glu-Tyr-Met-Phe-Cys-Ala330-Gly-Tyr-Ser-Asp-Gly-Ser-Lys-Asp-Ser-Cys340-Lys-Gly-Asp-Ser-Gly-Gly-Pro-His-Ala-Thr350-His-Tyr-Arg-Gly-Thr-Trp-Tyr-Leu-Thr-Gly360-Ile-Val-Ser-Trp-Gly-Gln-Gly-Cys-Ala-Thr370-Val-Gly-His-Phe-Gly-Val-Tyr-Thr-Arg-Val380-Ser-Gln-Tyr-Ile-Glu-Trp-Leu-Gln-Lys-Leu390-Met-Arg-Ser-Glu-Pro-Arg-Pro-Gly-Val-Leu400-Leu-Arg-Ala-Pro-Phe-Pro406
ATGGTCTCCCAGGCCCTCAGGCTCCTCTGCCTTCTGCTTGGGCTTCAGGGCTGCCTGGCTGCAGTCTTCGTAACCCAGGAGGAAGCCCAAGGCGTCCTGCACCGGCGCCGGCGCGCCAACGCGTTCCTGGAGGAGCTGCGGCCGGGCTCCCTGGAGAGGGAGTGCAAGGAGGAGCAGTGCTCCTTCGAGGAGGCCCGGGAGATCTTCAAGGACGCGGAGAGGACGAAGCTGTTCTGGATTTCTTACAGTGATGGGGACCAGTGTGCCTCAAGTCCATGCCAGAATGGGGGCTCCTGCAAGGACCAGCTCCAGTCCTATATCTGCTTCTGCCTCCCTGCCTTCGAGGGCCGGAACTGTGAGACGCACAAGGATGACCAGCTGATCTGTGTGAACGAGAACGGCGGCTGTGAGCAGTACTGCAGTGACCACACGGGCACCAAGCGCTCCTGTCGGTGCCACGAGGGGTACTCTCTGCTGGCAGACGGGGTGTCCTGCACACCCACAGTTGAATATCCATGTGGAAAAATACCTATTCTAGAAAAAAGAAATGCCAGCAAACCCCAAGGCCGAATTGTGGGGGGCAAGGTGTGCCCCAAAGGGGAGTGTCCATGGCAGGTCCTGTTGTTGGTGAATGGAGCTCAGTTGTGTGGGGGGACCCTGATCAACACCATCTGGGTGGTCTCCGCGGCCCACTGTTTCGACAAAATCAAGAACTGGAGGAACCTGATCGCGGTGCTGGGCGAGCACGACCTCAGCGAGCACGACGGGGATGAGCAGAGCCGGCGGGTGGCGCAGGTCATCATCCCCAGCACGTACGTCCCGGGCACCACCAACCACGACATCGCGCTGCTCCGCCTGCACCAGCCCGTGGTCCTCACTGACCATGTGGTGCCCCTCTGCCTGCCCGAACGGACGTTCTCTGAGAGGACGCTGGCCTTCGTGCGCTTCTCATTGGTCAGCGGCTGGGGCCAGCTGCTGGACCGTGGCGCCACGGCCCTGGAGCTCATGGTCCTCAACGTGCCCCGGCTGATGACCCAGGACTGCCTGCAGCAGTCACGGAAGGTGGGAGACTCCCCAAATATCACGGAGTACATGTTCTGTGCCGGCTACTCGGATGGCAGCAAGGACTCCTGCAAGGGGGACAGTGGAGGCCCACATGCCACCCACTACCGGGGCACGTGGTACCTGACGGGCATCGTCAGCTGGGGCCAGGGCTGCGCAACCGTGGGCCACTTTGGGGTGTACACCAGGGTCTCCCAGTACATCGAGTGGCTGCAAAAGCTCATGCGCTCAGAGCCACGCCCAGGAGTCCTCCTGCGAGCCCCATTTCCCTAG
Ser1-Gly-Thr-Thr-Asn-Thr-Val-Ala-Ala-Tyr10-Asn-Leu-Thr-Trp-Lys-Ser-Thr-Asn-Phe-Lys20-Thr-Ile-Leu-Glu-Trp-Glu-Pro-Lys-Pro-Val30-Asn-Gln-Val-Tyr-Thr-Val-Gln-Ile-Ser-Thr40-Lys-Ser-Gly-Asp-Trp-Lys-Ser-Lys-Cys-Phe50-Tyr-Thr-Thr-Asp-Thr-Glu-Cys-Asp-Leu-Thr60-Asp-Glu-Ile-Val-Lys-Asp-Val-Lys-Gln-Thr70-Tyr-Leu-Ala-Arg-Val-Phe-Ser-Tyr-Pro-Ala80-Gly-Asn-Val-Glu-Ser-Thr-Gly-Ser-Ala-Gly90-Glu-Pro-Leu-Tyr-Glu-Asn-Ser-Pro-Glu-Phe100-Thr-Pro-Tyr-Leu-Glu-Thr-Asn-Leu-Gly-Gln110-Pro-Thr-Ile-Gln-Ser-Phe-Glu-Gln-Val-Gly120-Thr-Lys-Val-Asn-Val-Thr-Val-Glu-Asp-Glu130-Arg-Thr-Leu-Val-Arg-Arg-Asn-Asn-Thr-Phe140-Leu-Ser-Leu-Arg-Asp-Val-Phe-Gly-Lys-Asp150-Leu-Ile-Tyr-Thr-Leu-Tyr-Tyr-Trp-Lys-Ser160-Ser-Ser-Ser-Gly-Lys-Lys-Thr-Ala-Lys-Thr170-Asn-Thr-Asn-Glu-Phe-Leu-Ile-Asp-Val-Asp180-Lys-Gly-Glu-Asn-Tyr-Cys-Phe-Ser-Val-Gln190-Ala-Val-Ile-Pro-Ser-Arg-Thr-Val-Asn-Arg200-Lys-Ser-Thr-Asp-Ser-Pro-Val-Glu-Cys-Met210-Gly-Gln-Glu-Lys-Gly-Glu-Phe-Arg-Glu219
ATGGAGACCCCTGCCTGGCCCCGGGTCCCGCGCCCCGAGACCGCCGTCGCTCGGACGCTCCTGCTCGGCTGGGTCTTCGCCCAGGTGGCCGGCGCTTCAGGCACTACAAATACTGTGGCAGCATATAATTTAACTTGGAAATCAACTAATTTCAAGACAATTTTGGAGTGGGAACCCAAACCCGTCAATCAAGTCTACACTGTTCAAATAAGCACTAAGTCAGGAGATTGGAAAAGCAAATGCTTTTACACAACAGACACAGAGTGTGACCTCACCGACGAGATTGTGAAGGATGTGAAGCAGACGTACTTGGCACGGGTCTTCTCCTACCCGGCAGGGAATGTGGAGAGCACCGGTTCTGCTGGGGAGCCTCTGTATGAGAACTCCCCAGAGTTCACACCTTACCTGGAGACAAACCTCGGACAGCCAACAATTCAGAGTTTTGAACAGGTGGGAACAAAAGTGAATGTGACCGTAGAAGATGAACGGACTTTAGTCAGAAGGAACAACACTTTCCTAAGCCTCCGGGATGTTTTTGGCAAGGACTTAATTTATACACTTTATTATTGGAAATCTTCAAGTTCAGGAAAGAAAACAGCCAAAACAAACACTAATGAGTTTTTGATTGATGTGGATAAAGGAGAAAACTACTGTTTCAGTGTTCAAGCAGTGATTCCCTCCCGAACAGTTAACCGGAAGAGTACAGACAGCCCGGTAGAGTGTATGGGCCAGGAGAAAGGGGAATTCAGAGAATAA
sTF(1−219) W45C、sTF(1−219)D58C、sTF(1−219)G109C、及びsTF(1−219)P206CをコードするDNAの構築 − 製造者(Stratagene, La Jolla, CA)の指示書に従って、表3に挙げたプライマー対(順方向及び逆方向)と鋳型としてpHOJ356を用いるQuickChange(登録商標)部位特異的突然変異誘発によって、sTF(1−219)の変異体を構築した。全てのクローン化配列の正確な同一性はDNA配列決定によって証明される。
sTF(1−209)及びsTF(1−209)V207CをコードするDNAの構築 − そのシグナルペプチドを含むTF(1−209)のDNAコード配列を、製造者の推奨に従ってExpand・High・Fidelity・PCRシステム(Roche Diagnostics Corporation, Indianapolis, IN)及びプライマーoHOJ152−f(配列は表1に示す)及びoHOJ178−r(配列は表5に示す)を使用し、フランキングNheI及びXhoI制限部位を導入するPCRによって、ヒト全脳cDNAライブラリー(Marathon-ready cDNA; Clontech Laboratories Inc., Mountain View, CA)から増幅させる。精製したPCR産物をNheI及びXhoIで切断し、ついで対応するpCI−neo(Promega, Madison, WI)の対応する部位に結合させる。ついで、得られたプラスミドは、製造者(Stratagene, La Jolla, CA)の指示書に従って、QuickChange(登録商標)部位特異的突然変異誘発及びプライマーoHOJ179−f及びoHOJ179−r(配列は表5に示す)を使用してのsTF(1−209)V207Cの構築のためのテンプレートとなる。全てのクローン化配列の正確な同一性はDNA配列決定によって証明される。
Claims (8)
- VII因子ポリペプチドと組織因子ポリペプチドの共有結合複合体であって、VII因子ポリペプチドと組織因子ポリペプチドが、次のアミノ酸セット:
FVII−Phe40 − sTF−Val207;
FVII−Gln64 − sTF−Gly109;
FVII−Phe275 − sTF−Trp45
の少なくとも一つの間での一又は複数の直接結合によって共有結合されている複合体。 - VII因子ポリペプチドが機能的に活性なVII因子ポリペプチドである請求項1に記載の複合体。
- VII因子ポリペプチドのタンパク分解活性が、インビトロタンパク分解アッセイによって測定して、遊離の天然(野生型)VIIa因子の活性の少なくとも2倍である請求項2に記載の複合体。
- アミノ酸のセットが一又は複数のシステインアミノ酸を含む請求項1から3の何れか一項に記載の複合体。
- 該セットのそれぞれが1個のシステインアミノ酸を含む請求項4に記載の複合体。
- 該セットのそれぞれが2個のシステインアミノ酸を含む請求項4に記載の複合体。
- 少なくとも一つの直接結合が2個のシステインアミノ酸間にある請求項1から6の何れか一項に記載の複合体。
- 請求項1から7の何れか一項に記載のVII因子ポリペプチドと組織因子ポリペプチドの複合体の製造方法であって、a)(i)VII因子ポリペプチドをコードする核酸分子と該核酸分子に作用可能に結合した発現制御領域を含む発現ベクターと(ii)組織因子ポリペプチドをコードする核酸分子と該核酸分子に作用可能に結合した発現制御領域を含む発現ベクターとを細胞に形質移入し、b)形質移入細胞を、VII因子ポリペプチドと組織因子ポリペプチドの発現条件下で培養し、c)発現された複合体を単離することを含む方法。
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EP06112358.4 | 2006-04-07 | ||
EP06112358 | 2006-04-07 | ||
PCT/EP2007/053105 WO2007115953A1 (en) | 2006-04-07 | 2007-03-30 | Covalent factor vii-tissue factor complex |
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EP (1) | EP2007417B1 (ja) |
JP (1) | JP5759102B2 (ja) |
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EP2127640A1 (en) | 2008-05-27 | 2009-12-02 | Koninklijke Philips Electronics N.V. | Azide modified proteins |
KR20120039658A (ko) | 2009-06-24 | 2012-04-25 | 코닌클리즈케 필립스 일렉트로닉스 엔.브이. | 마이크로컨트롤러를 프로그래밍하기 위한 방법 및 장치 |
WO2011092242A1 (en) * | 2010-01-28 | 2011-08-04 | Novo Nordisk Health Care Ag | Factor vii fusion polypeptides |
WO2013143876A1 (en) * | 2012-03-30 | 2013-10-03 | Novo Nordisk A/S | Fviia-stf complexes exhibiting exosite-mediated super activity |
US10202595B2 (en) | 2012-06-08 | 2019-02-12 | Bioverativ Therapeutics Inc. | Chimeric clotting factors |
CN104519897A (zh) | 2012-06-08 | 2015-04-15 | 比奥根艾迪克Ma公司 | 促凝血化合物 |
EP2906693A1 (en) | 2012-10-15 | 2015-08-19 | Novo Nordisk Health Care AG | Coagulation factor vii polypeptides |
WO2017027545A1 (en) * | 2015-08-12 | 2017-02-16 | Cell Machines, Inc. | Methods and compositions related to long half-life coagulation complexes |
EP3956359A1 (en) | 2019-04-17 | 2022-02-23 | Novo Nordisk A/S | Bispecific antibodies |
CN114836503B (zh) * | 2022-04-29 | 2023-08-25 | 江苏祈瑞医药科技有限公司 | 一组具有肝损伤保护作用的乳清蛋白肽、高f值寡肽及其制备方法和应用 |
CN117860874B (zh) * | 2024-03-12 | 2024-07-02 | 正大天晴药业集团南京顺欣制药有限公司 | 重组人凝血因子VIIa的药物组合物 |
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US5788965A (en) * | 1991-02-28 | 1998-08-04 | Novo Nordisk A/S | Modified factor VII |
WO1997020939A1 (en) * | 1995-12-01 | 1997-06-12 | Genentech, Inc. | TISSUE FACTOR - KUNITZ DOMAIN FUSION PROTEINS AS FACTOR VIIa INHIBITORS |
IL132560A0 (en) * | 1997-05-02 | 2001-03-19 | Genentech Inc | A method for making multispecific antibodies having heteromultimeric and common components |
AU783512B2 (en) * | 2000-02-11 | 2005-11-03 | Bayer Healthcare Llc | Factor VII or VIIa-like molecules |
CA2441986A1 (en) * | 2001-03-27 | 2002-10-03 | Massachusetts Institute Of Technology | Methods and products related to fgf dimerization |
EP1432794B1 (en) * | 2001-09-27 | 2011-11-09 | Novo Nordisk Health Care AG | Human coagulation factor vii polypeptides |
EP2348043A1 (en) * | 2001-10-02 | 2011-07-27 | Genentech, Inc. | APO-2 ligand variants and uses thereof |
AU2003225255B2 (en) * | 2002-05-01 | 2008-07-31 | Bayer Schering Pharma Aktiengesellschaft | Novel tissue factor targeted antibodies as anticoagulants |
EP1523504A2 (en) * | 2002-07-12 | 2005-04-20 | Novo Nordisk A/S | A tissue factor binding immunoconjugate comprising factor viia |
EP1673453A2 (en) * | 2003-10-07 | 2006-06-28 | Novo Nordisk Health Care AG | Hybrid molecules having factor vii/viia activity |
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EP2007417A1 (en) | 2008-12-31 |
CN101466400A (zh) | 2009-06-24 |
ES2353084T3 (es) | 2011-02-25 |
CN101466400B (zh) | 2014-06-25 |
JP2009532048A (ja) | 2009-09-10 |
EP2007417B1 (en) | 2010-10-20 |
DE602007009956D1 (de) | 2010-12-02 |
US20110040073A1 (en) | 2011-02-17 |
ATE485052T1 (de) | 2010-11-15 |
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