[發明專利]用于糖尿病治療的蛋白、蛋白綴合物及其應用

有效

• 申請號: 201380018643.2
• 申請日: 2013-05-17
• 公開（公告）號: CN104302772B
• 公開（公告）日: 2017-11-10
• 發明（設計）人: 秦樹林
• 申請（專利權）人: 愛德迪安（北京）生物技術有限公司
• 主分類號: C12N15/62
• 分類號: C12N15/62;C07K19/00;C07K14/545;C07K14/605;C07K14/595;A61K38/16;A61K38/30;A61K38/26;A61P3/10
• 代理公司: 北京邦信陽專利商標代理有限公司11012
• 代理人: 黃澤雄,陳悅軍

1.一種融合蛋白，所述融合蛋白由白介素-1受體拮抗蛋白與另一種多肽連接而成，另一種多肽是GLP-1受體結合多肽；

所述融合蛋白的結構為：GLP-1受體結合多肽-Lj-白介素-1受體拮抗蛋白；

所述融合蛋白的序列為：

HX_G2EGTFTSDX_G10SX_G12YLEX_G16QAAX_G20EFIAWLVX_G28GRX_G31-L_j-X_IL0RPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVX_IL84ITDLSENRKQDKRFAFIRSDSGPTTSFESAAX_IL116PGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE，其中，X_G2為G，X_G10為V或C，X_G12為S，X_G16為E，X_G20為K或C，X_G28為K或C，X_G31為G或C或PSSGAPPPS，L_j為(GGGGS)_n，n為0、1、2或3，X_IL0為缺失，X_IL84為N或C，X_IL116為C或S。

2.根據權利要求1所述的融合蛋白，選自化合物G-18～G-29、G-40～G-43、G-45和G-47～G-50，其中，各化合物的結構如下：

G-18：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRGRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAACPGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE；

G-19：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAACPGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE；

G-20：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRGGGGSGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAACPGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE；

G-21：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRGGGGSGGGGSGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAACPGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE；

G-22：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRPSSGAPPPSGGGGSGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAACPGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE；

G-23：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRGRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAAC(S-CH₂-CONH-PEG20K)PGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE(S是半胱氨酸側鏈的硫原子)；

G-24：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAAC(S-CH₂-CONH-PEG20K)PGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE(S是半胱氨酸側鏈的硫原子)；

G-25：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVC[S-馬來酰亞胺-(CH₂)₄-NH-(N^α-(HOOC(CH₂)₁₄CO)-γ-Glu)]ITDLSENRKQDKRFAFIRSDSGPTTSFESAASPGWFLCTAME ADQPVSLTNMPDEGVMVTKFYFQEDE(S是半胱氨酸側鏈的硫原子)；

G-26：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRGGGGSGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAAC(S-CH₂-CONH-PEG20K)PGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE(S是半胱氨酸側鏈的硫原子)；

G-27：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRGGGGSGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVC[S-馬來酰亞胺-(CH₂)₄-NH-(N^α-(HOOC(CH₂)₁₄CO)-γ-Glu)]ITDLSENRKQDKRFAFIRSDSGPTTSFESAASPGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE(S是半胱氨酸側鏈的硫原子)；

G-28：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRGGGGSGGGGSGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAAC(S-CH₂-CONH-PEG20K)PGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE(S是半胱氨酸側鏈的硫原子)；

G-29：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRPSSGAPPPSGGGGSGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAAC[S-馬來酰亞胺-(CH₂)₄-NH-(N^α-(HOOC(CH₂)₁₄CO)-γ-Glu)]PGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE(S是半胱氨酸側鏈的硫原子)；

G-40：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRC(S-馬來酰亞胺-(CH₂)₁₅-COOH)GGGGSGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAASPGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE(S是半胱氨酸側鏈的硫原子)；

G-41：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRC[S-CH₂-CONH-(CH₂CH₂O)₂-(CH₂)₂-NH-(N^α-(HOOC(CH₂)₁₄CO)-γ-Glu)]GGGGSGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAASPGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE(S是半胱氨酸側鏈的硫原子)；

G-42：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRGGGGSGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVC(S-馬來酰亞胺-(CH₂)₁₇-COOH)ITDLSENRKQDKRFAFIRSDSGPTTSFESAASPGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE(S是半胱氨酸側鏈的硫原子)；

G-43：

HGEGTFTSDVSSYLEEQAAC[S-CH₂-CONH-(CH₂CH₂O)₂-(CH₂)₂-NH-(N^α-(HOOC(CH₂)₁₄CO)-γ-Glu)]EFIAWLVKGRGGGGSGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAASPGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE(S是半胱氨酸側鏈的硫原子)；

G-45：

HGEGTFTSDC[S-CH₂-CONH-(CH₂CH₂O)₂-(CH₂)₂-NH-(N^α-(HOOC(CH₂)₁₄CO-γ-Glu-N-

γ-Glu)]SSYLEEQAAKEFIAWLVKGRGGGGSGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAASPGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE(S是半胱氨酸側鏈的硫原子)；

G-47：

HGEGTFTSDVSSYLEEQAAKEFIAWLVC[S-CH₂-CONH-(CH₂CH₂O)₂-(CH₂)₂-NH-(N^α-(HOOC(CH₂)₁₄CO)-γ-Glu)]GRGGGGSGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAASPGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE(S是半胱氨酸側鏈的硫原子)；

G-48：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRGGGGSGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVC[S-CH₂-CONH-(CH₂CH₂O)₄-(CH₂)₂-NH-(N^α-(HOOC(CH₂)₁₆CO)-γ-Glu)]ITDLSENRKQDKRFAFIRSDSGPTTSFESAASPGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE(S是半胱氨酸側鏈的硫原子)；

G-49：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRGGGGSGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAAC(S-馬來酰亞胺-(CH₂)₁₅-COOH)PGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE(S是半胱氨酸側鏈的硫原子)；

G-50：

HGEGTFTSDVSSYLEEQAAKEFIAWLVKGRGGGGSGGGGSRPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLSCVKSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAAC[S-CH₂-CONH-(CH₂CH₂O)₂-(CH₂)₂-NH-(N^α-(HOOC(CH₂)₁₄CO)-γ-Glu)]PGWFLCTAMEADQPVSLTNMPDEGVMVTKFYFQEDE(S是半胱氨酸側鏈的硫原子)。