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    Process for producing peptides in E. coli
    6.
    发明授权
    Process for producing peptides in E. coli 失效
    在大肠杆菌中生产肽的方法

    公开(公告)号：US5670340A

    公开(公告)日：1997-09-23

    申请号：US352179

    申请日：1994-12-05

    申请人： Masayuki Yabuta ,  Yuji Suzuki ,  Kazuhiro Ohsuye ,  Takehiro Oshima ,  Seiko Onai ,  Koji Magota ,  Shoji Tanaka

    发明人： Masayuki Yabuta ,  Yuji Suzuki ,  Kazuhiro Ohsuye ,  Takehiro Oshima ,  Seiko Onai ,  Koji Magota ,  Shoji Tanaka

    IPC分类号： C12N15/09 ,  C07K14/58 ,  C07K14/585 ,  C12N9/38 ,  C12N15/16 ,  C12N15/62 ,  C12N15/67 ,  C12N15/70 ,  C12P21/02 ,  C12P21/06 ,  C12R1/19 ,  C12P21/00

    CPC分类号： C07K14/58 ,  C07K14/585 ,  C12N15/62 ,  C12N9/2471 ,  C12Y302/01023 ,  C07K2319/00 ,  C07K2319/23 ,  C07K2319/43 ,  C07K2319/50 ,  C07K2319/75 ,  C07K2319/90

    摘要： The present invention is a process to express a target peptide in a large amount and accumulate the target peptide in host cells in the form of inclusion bodies. The process comprises: A) culturing host cells transformed with a plasmid able to express a gene coding for a fusion protein represented in the formula A--L--B, wherein B is a target peptide, A is a protective peptide comprising a 90-210 amino acid fragment E. coli .beta.-galactosidase, and L is a linker peptide positioned between the C-terminus of the protective peptide and the N-terminus of the target peptide and selected so that when the fusion protein is treated by an enzyme or chemical substance, the target peptide is separated, and wherein the protective peptide and linker peptide are selected so that the isoelectric point of the fusion protein in between 4.9 and 6.9; B) obtaining an insoluble fraction comprising inclusion bodies by homogenization of th cultured transformed cells; C) solubilizing the fusion protein in the inclusion bodies by treatment of the insoluble fraction with a solubilizing agent; and, D) cleaving the peptide bond between the C-terminus of the linker peptide and the N-terminus of the target peptide of the solubilized fusion protein to release the target peptide from the other peptides followed by purification of the target peptide.

    摘要翻译： 本发明是大量表达目标肽并以包涵体的形式在宿主细胞中聚集靶肽的方法。 该方法包括：A）培养用能够表达编码式ALB中表示的融合蛋白的基因的质粒转化的宿主细胞，其中B是靶肽，A是包含90-210个氨基酸片段E的保护肽 大肠杆菌β-半乳糖苷酶，L是位于保护肽的C-末端和靶肽的N末端之间的连接肽，并且被选择使得当融合蛋白被酶或化学物质处理时，靶 分离肽，并且其中选择保护性肽和接头肽，使得融合蛋白的等电点在4.9和6.9之间; B）通过均质培养的转化细胞获得包含包含体的不溶性级分; C）通过用增溶剂处理不溶性级分将融合蛋白溶解在包涵体中; 并且D）切割接头肽的C末端和溶解的融合蛋白的靶肽的N末端之间的肽键，从其他肽释放靶肽，然后纯化目标肽。

    Precursor of a C-terminal amidated calcitonin
    7.
    发明授权
    Precursor of a C-terminal amidated calcitonin 失效
    C-末端酰胺化降钙素前体

    公开(公告)号：US5252482A

    公开(公告)日：1993-10-12

    申请号：US610312

    申请日：1990-11-05

    申请人： Shoji Tanaka ,  Kazuhiro Ohsuye ,  Ichiro Kubota ,  Norio Ohnuma ,  Teruhisa Noguchi

    发明人： Shoji Tanaka ,  Kazuhiro Ohsuye ,  Ichiro Kubota ,  Norio Ohnuma ,  Teruhisa Noguchi

    IPC分类号： A61K38/00 ,  C07K14/585 ,  C12N1/21 ,  C12N15/16 ,  C12N15/70

    CPC分类号： C07K14/585 ,  A61K38/00

    摘要： A precursor of a C-terminal amidated peptide represented by the general formula P-X-Gly-Y.sub.n, wherein P is a peptide residue. X is an amino acid residue the C terminal of which (on the Gly side) can be converted in vivo to a --CONH.sub.2 group. Gly is a glycine residue, Y is a basic amino acid residue, n is an interger of 2 to 4 and a further amino acid residue other than Y or a peptide residue may be attached to Y.sub.n, is produced by a gene engineering technology. The precursor exhibits in vivo physiological activity like the C-terminal amidated peptide.

    摘要翻译： 由通式P-X-Gly-Yn表示的C末端酰胺化肽的前体，其中P是肽残基。 X是其末端（Gly侧）可以在体内转化为-CONH 2基团的氨基酸残基。 Gly是甘氨酸残基，Y是碱性氨基酸残基，n是2至4的整数，并且除Y之外的另外的氨基酸残基或肽残基可以连接到Yn上，是通过基因工程技术产生的。 前体表现出体内生理活性，如C-末端酰胺化肽。

    Human CNP gene and precursor protein
    8.
    发明授权
    Human CNP gene and precursor protein 失效
    人CNP基因和前体蛋白

    公开(公告)号：US6034231A

    公开(公告)日：2000-03-07

    申请号：US765830

    申请日：1991-09-26

    申请人： Shoji Tanaka ,  Kayoko Fuchimura ,  Yasunori Tawaragi ,  Hisayuki Matsuo ,  Kenji Kanagawa ,  Naoto Minamino

    发明人： Shoji Tanaka ,  Kayoko Fuchimura ,  Yasunori Tawaragi ,  Hisayuki Matsuo ,  Kenji Kanagawa ,  Naoto Minamino

    IPC分类号： C12N15/09 ,  A61K38/00 ,  A61K38/22 ,  A61P7/10 ,  C07K14/00 ,  C07K14/575 ,  C07K14/58 ,  C12N15/16 ,  C07H17/00

    CPC分类号： C07K14/58 ,  A61K38/00

    摘要： The gene of a human CNP (human C-type natriuretic peptide), as well as the hCNP precursor protein that is encoded by said gene. The hCNP precursor is represented by the following amino acid sequence.Met His Leu Ser Gln Leu Leu Ala Cys Ala - Leu Leu Leu Thr Leu Leu Ser Leu Arg Pro - Ser Glu Ala Lys Pro Gly Ala Pro Pro Lys - Val Pro Arg Thr Pro Pro Ala Glu Glu Leu - Ala Glu Pro Gln Ala Ala Gly Gly Gly Gln - Lys Lys Gly Asp Lys Ala Pro Gly Gly Gly - Gly Ala Asn Leu Lys Gly Asp Arg Ser Arg - Leu Leu Arg Asp Leu Arg Val Asp Thr Lys - Ser Arg Ala Ala Trp Ala Arg Leu Leu Gln - Glu His Pro Asn Ala Arg Lys Tyr Lys Gly - Ala Asn Lys Lys Gly Leu Ser Lys Gly Cys - Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser - Met Ser Gly Leu Gly Cys. The hCNP precursor and its derivatives are novel and have natriuretic and hypotensive activities.

    摘要翻译： 人CNP（人C型利钠肽）的基因以及由所述基因编码的hCNP前体蛋白。 hCNP前体由以下氨基酸序列表示。 - Met His Leu Ser Gln Leu Leu Ala Cys Ala - - Leu Leu Leu Leu Leu Leu Leu Ser Leu Arg Pro - - Ser Glu Ala Lys Pro Gly Ala Pro Pro Lys - - Val Pro Arg Thr Pro Pro Ala Glu Glu Leu - - Ala Glu Pro Gln Ala Ala Gly Gly Gly Gln - - Lys Lys Gly Asp Lys Ala Pro Gly Gly Gly - - Gly Ala Asn Leu Lys Gly Asp Arg Ser Arg - - Leu Leu Arg Asp Leu Arg Val Asp Thr Lys - - Ser Arg Ala Ala Trp Ala Arg Leu Leu Gln - - Glu His Pro Asn Ala Arg Lys Tyr Lys Gly - - Ala Asn Lys Lys Gly Leu Ser Lys Gly Cys - Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser - - Met Ser Gly Leu Gly Cys。 - hCNP前体及其衍生物是新颖的，具有钠尿和降血压活性。

    CNP analog peptides and their use
    9.
    发明授权
    CNP analog peptides and their use 失效
    CNP类似物及其用途

    公开(公告)号：US5434133A

    公开(公告)日：1995-07-18

    申请号：US828450

    申请日：1992-01-31

    申请人： Shoji Tanaka ,  Yoshiharu Minamitake ,  Yasuo Kitajima ,  Mayumi Furuya ,  Hisayuki Matsuo

    发明人： Shoji Tanaka ,  Yoshiharu Minamitake ,  Yasuo Kitajima ,  Mayumi Furuya ,  Hisayuki Matsuo

    IPC分类号： A61K38/22 ,  A61K38/00 ,  A61P43/00 ,  C07K7/08 ,  C07K14/575 ,  C07K14/58 ,  C12N15/16 ,  C07K14/00 ,  A61K38/16 ,  A61K38/17 ,  C07K14/435

    CPC分类号： C07K14/58 ,  A61K38/00 ,  Y02P20/55

    摘要： Novel peptides represented by the general formula: ##STR1## and physiologically acceptable acid addition salts thereof; where (A) represents H-, H-Gly, H-Lys-Gly, H-Ser-Lys-Gly, H-Leu-Ser-Lys-Gly, H-Gly-Leu-Ser-Lys-Gly, H-ser, H-Ser-Ser, H-Arg-Ser-Ser, H-Arg-Arg-Ser-Ser, H-Leu-Arg-Arg-Ser-Ser, H-Ser-Leu-Arg-Arg-Ser-Ser;(B) represents H-Cys or Pmp;(C) represents Phe-, pCl-Phe, pF-Phe, pNO.sub.2 -Phe or Cha;(D) represents Ile, Val, Aib, tLeu, Gly or Leu;(E) represents Lys or Arg;(F) represents Ile, Leu or Met;(G) represents Ser or Ala;(H) represents Met or Gln;(I) represents --OH, -Asn-OH, -Asn-Ser-OH, -Asn-Ser-Phe-OH, -Asn-Ser-Phe-Arg-OH or -Asn-Ser-Phe-Arg-Tyr-OH; and the symbol " . . . " represents a disulfide bond;provided that 1) .alpha.-hANP, 2) .alpha.-hANP (7-28) and 3) CNP-22 are excluded from the scope of that general formula.Also disclosed are agents for suppressing the growth of vascular smooth muscle cells that contains those peptides as effective ingredients.

    摘要翻译： 由通式表示的新肽：其加成盐; 其中（A）表示H，H-Gly，H-Lys-Gly，H-Ser-Lys-Gly，H-Leu-Ser-Lys-Gly，H-Gly-Leu-Ser-Lys-Gly， Ser，H-Ser-Ser，H-Arg-Ser-Ser，H-Arg-Arg-Ser-Ser，H-Leu-Arg-Arg-Ser-Ser，H-Ser-Leu-Arg-Arg- 塞尔 （B）代表H-Cys或Pmp; （C）表示Phe-，pCl-Phe，pF-Phe，pNO2-Phe或Cha; （D）代表Ile，Val，Aib，tLeu，Gly或Leu; （E）表示Lys或Arg; （F）表示Ile，Leu或Met; （G）表示Ser或Ala; （H）表示Met或Gln; （I）表示-OH，-Asn-OH，-Asn-Ser-OH，-Asn-Ser-Phe-OH，-Asn-Ser-Phe-Arg-OH或-Asn-Ser-Phe-Arg-Tyr- 哦; 符号“......”代表二硫键; 条件是1）α-hANP，2）α-hANP（7-28）和3）CNP-22被排除在该通式的范围之外。 还公开了抑制含有那些肽作为有效成分的血管平滑肌细胞生长的药剂。

    Porcine CNP gene and precursor protein
    10.
    发明授权
    Porcine CNP gene and precursor protein 失效
    猪CNP基因和前体蛋白

    公开(公告)号：US6020168A

    公开(公告)日：2000-02-01

    申请号：US728220

    申请日：1991-07-12

    申请人： Hisayuki Matsuo ,  Kenji Kangawa ,  Naoto Minamino ,  Shoji Tanaka ,  Kayoko Fuchimura ,  Yasunori Tawaragi

    发明人： Hisayuki Matsuo ,  Kenji Kangawa ,  Naoto Minamino ,  Shoji Tanaka ,  Kayoko Fuchimura ,  Yasunori Tawaragi

    IPC分类号： C12N15/09 ,  A61K38/00 ,  C07H21/04 ,  C07K14/575 ,  C07K14/58 ,  C12N15/16 ,  C12P21/02 ,  C12R1/19 ,  C12P21/06 ,  C07H17/00 ,  C07K14/00

    CPC分类号： C07K14/58 ,  A61K38/00

    摘要： The gene and cDNA of a procine derived CNP (C-type natriuretic peptide), and a procine derived CNP precursor protein and derivatives thereof are disclosed. The proceine derivative CNP precursor is represented by the following amino acid sequence:Met His Leu Ser Gln Leu Leu Ala Cys Ala - Leu Leu Leu Thr Leu Leu Ser Leu Arg Pro - Ser Glu Ala Lys Pro Gly Ala Pro Pro Lys - Val Pro Arg Thr Pro Pro Gly Glu Glu Val - Ala Glu Pro Gln Ala Ala Gly Gly Gly Gln - Lys Lys Gly Asp Lys Thr Pro Gly Gly Gly - Gly Ala Asn Leu Lys Gly Asp Arg Ser Arg - Leu Leu Arg Asp Leu Arg Val Asp Thr Lys - Ser Arg Ala Ala Trp Ala Arg Leu Leu His - Glu His Pro Asn Ala Arg Lys Tyr Lys Gly - Gly Asn Lys Lys Gly Leu Ser Lys Gly Cys - Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser These derivatives are novel and have natriuretic and hypotensive activities.

    摘要翻译： 公开了源自CNP（C型利尿钠肽）的基因和cDNA，以及源自CNP前体蛋白及其衍生物的衍生物。 程序衍生物CNP前体由以下氨基酸序列表示： - Met His Leu Ser Gln Leu Leu Ala Cys Ala - - Leu Leu Leu Thr Leu Leu Ser Leu Arg Pro - Ser Glu Ala Lys Pro Gly Ala Pro Pro Lys - - Val Pro Arg Thr Pro Pro Gly Glu Glu Val - - Ala Glu Pro Gln Ala Ala Gly Gly Gly Gln - - Lys Lys Gly Asp Lys Thr Pro Gly Gly Gly - - Gly Ala Asn Leu Lys Gly Asp Arg Ser Arg - - Leu Leu Arg Asp Leu Arg Val Asp Thr Lys - - Ser Arg Ala Ala Trp Ala Arg Leu Leu His - - Glu His Pro Asn Ala Arg Lys Tyr Lys Gly - - Gly Asn Lys Lys Gly Leu Ser Lys Gly Cys - Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser - - Met Ser Gly Leu Gly Cys。 - 这些衍生物是新颖的，具有钠尿和降血压活性。