公开(公告)号：US20110028691A1

公开(公告)日：2011-02-03

申请号：US12900214

申请日：2010-10-07

申请人： Clifford Charles SHONE ,  Keith Alan FOSTER ,  John CHADDOCK ,  Philip MARKS ,  J. Mark SUTTON ,  Patrick STANCOMBE ,  Jonathan WAYNE

发明人： Clifford Charles SHONE ,  Keith Alan FOSTER ,  John CHADDOCK ,  Philip MARKS ,  J. Mark SUTTON ,  Patrick STANCOMBE ,  Jonathan WAYNE

IPC分类号： C07K14/33 ,  C07H21/04

CPC分类号： A61K39/08 ,  A61K38/00 ,  A61K39/00 ,  A61K47/6415 ,  A61K47/646 ,  A61K2039/505 ,  A61K2039/53 ,  A61K2039/627 ,  C07K14/33 ,  C07K2319/00 ,  C07K2319/20 ,  C07K2319/23 ,  C07K2319/50 ,  C07K2319/55 ,  C07K2319/705 ,  C07K2319/75 ,  C12N9/52 ,  C12N15/62 ,  Y10S530/825

摘要： A single polypeptide is provided which comprises first and second domains. The first domain enables the polypeptide to cleave one or more vesicle or plasma-membrane associated proteins essential to exocytosis, and the second domain enables the polypeptide to be translocated into a target cell or increases the solubility of the polypeptide, or both. The polypeptide thus combines useful properties of a clostridial toxin, such as a botulinum or tetanus toxin, without the toxicity associated with the natural molecule. The polypeptide can also contain a third domain that targets it to a specific cell, rendering the polypeptide useful in inhibition of exocytosis in target cells. Fusion proteins comprising the polypeptide, nucleic acids encoding the polypeptide and methods of making the polypeptide are also provided. Controlled activation of the polypeptide, is possible and the polypeptide can be incorporated into vaccines and toxin assays.

摘要翻译： 提供了包含第一和第二结构域的单个多肽。 第一结构域使得多肽能够切割一种或多种与胞吐作用相关的囊泡或质膜相关的蛋白质，而第二结构域使得多肽能够转位到靶细胞中或者增加多肽的溶解性，或者两者。 因此，该多肽结合梭菌毒素如肉毒杆菌或破伤风毒素的有用性质，而不与天然分子相关的毒性。 多肽还可以含有将其靶向特定细胞的第三结构域，使得多肽可用于抑制靶细胞中的胞吐作用。 还提供了包含多肽的融合蛋白，编码多肽的核酸和制备多肽的方法。 多肽的受控活化是可能的，并且多肽可以并入疫苗和毒素测定中。

公开(公告)号：US20120149084A1

公开(公告)日：2012-06-14

申请号：US13354787

申请日：2012-01-20

申请人： Keith Alan FOSTER ,  John CHADDOCK ,  Philip MARKS ,  Patrick STANCOMBE ,  Lyndsey DUROSE

发明人： Keith Alan FOSTER ,  John CHADDOCK ,  Philip MARKS ,  Patrick STANCOMBE ,  Lyndsey DUROSE

IPC分类号： C12N9/96 ,  C12N15/63 ,  C12N15/62

CPC分类号： C12N9/50 ,  A61K38/00 ,  C07K14/33 ,  C07K2319/06 ,  C07K2319/50 ,  C12N15/62

摘要： The invention provides a single chain, polypeptide fusion protein, comprising: a non-cytotoxic protease, or a fragment thereof, which protease or protease fragment is capable of cleaving a protein of the exocytic fusion apparatus of a target cell; a Targeting Moiety that is capable of binding to a Binding Site on the target cell, which Binding Site is capable of undergoing endocytosis to be incorporated into an endocome within the target cell; a protease cleaving site at which site the fusion protein is cleavable by the protease, wherein the protease cleavage site is located between the non-cytotoxic protease or fragment thereof and the Targeting Moiety; and the translocation domain that is capable of translocating the protease or protease fragment from within an endosome, across the endosomal membrane and into the cytosol of the target cell.

摘要翻译： 本发明提供了一种单链多肽融合蛋白，其包含：非细胞毒性蛋白酶或其片段，所述蛋白酶或蛋白酶片段能够切割靶细胞的胞外融合装置的蛋白质; 能够结合靶细胞上的结合位点的靶向物质，所述结合位点能够经历内吞作用以掺入靶细胞内的内皮细胞; 蛋白酶切割位点，其中融合蛋白可被蛋白酶切割，其中蛋白酶切割位点位于非细胞毒性蛋白酶或其片段与靶向部位之间; 以及能够将位于内体内的蛋白酶或蛋白酶片段穿过体内膜并进入靶细胞的胞质溶胶的易位结构域。

公开(公告)号：US20110027256A1

公开(公告)日：2011-02-03

申请号：US12862948

申请日：2010-08-25

申请人： Keith FOSTER ,  John CHADDOCK ,  Philip MARKS ,  Patrick STANCOMBE ,  Kei Roger AOKI ,  Joseph FRANCIS ,  Lance STEWARD

发明人： Keith FOSTER ,  John CHADDOCK ,  Philip MARKS ,  Patrick STANCOMBE ,  Kei Roger AOKI ,  Joseph FRANCIS ,  Lance STEWARD

IPC分类号： A61K38/48 ,  C07H21/04 ,  A61P29/00 ,  C12N9/48 ,  C12N15/63

CPC分类号： C12N9/96 ,  A61K38/00 ,  A61K47/64 ,  C07K2319/01 ,  C07K2319/33 ,  C07K2319/50 ,  C12N9/50

摘要： A single chain, polypeptide fusion protein, comprising: a non-cytotoxic protease, or a fragment thereof, which protease or protease fragment is capable of cleaving a protein of the exocytic fusion apparatus of a nociceptive sensory afferent; a dynorphin Targeting Moiety that is capable of binding to a Binding Site on the nociceptive sensory afferent, which Binding Site is capable of undergoing endocytosis to be incorporated into an endosome within the nociceptive sensory afferent; a protease cleavage site at which site the fusion protein is cleavable by a protease, wherein the protease cleavage site is located between the non-cytotoxic protease or fragment thereof and the dynorphin Targeting Moiety; and a translocation domain that is capable of translocating the protease or protease fragment from within an endosome, across the endosomal membrane and into the cytosol of the nociceptive sensory afferent. Nucleic acid sequences encoding the polypeptide fusion proteins, methods of preparing same and uses thereof are also described.

公开(公告)号：US20120064059A1

公开(公告)日：2012-03-15

申请号：US13239573

申请日：2011-09-22

申请人： Keith FOSTER ,  John CHADDOCK ,  Philip MARKS ,  Patrick STANCOMBE ,  Kei Roger AOKI ,  Joseph FRANCIS ,  Lance STEWARD

发明人： Keith FOSTER ,  John CHADDOCK ,  Philip MARKS ,  Patrick STANCOMBE ,  Kei Roger AOKI ,  Joseph FRANCIS ,  Lance STEWARD

IPC分类号： A61K38/48 ,  A61P25/04 ,  C12N15/63 ,  C12N9/50 ,  C12N15/57

CPC分类号： C07K14/575 ,  A61K31/711 ,  A61K38/4893 ,  A61K47/64 ,  A61K47/6415 ,  C07K14/33 ,  C07K2319/00 ,  C07K2319/01 ,  C07K2319/33 ,  C12N9/6489 ,  C12N15/62 ,  C12Y304/24069

摘要： A single chain, polypeptide fusion protein, comprising: a non-cytotoxic protease, or a fragment thereof, which protease or protease fragment is capable of cleaving a protein of the exocytic fusion apparatus of a nociceptive sensory afferent; a Targeting Moiety that is capable of binding to a Binding Site on the nociceptive sensory afferent, which Binding Site is capable of undergoing endocytosis to be incorporated into an endosome within the nociceptive sensory afferent; a protease cleavage site at which site the fusion protein is cleavable by a protease, wherein the protease cleavage site is located between the non-cytotoxic protease or fragment thereof and the Targeting Moiety; a translocation domain that is capable of translocating the protease or protease fragment from within an endosome, across the endosomal membrane and into the cytosol of the nociceptive sensory afferent. Nucleic acids encoding the fusion proteins, methods of preparing same and uses thereof are also described.

摘要翻译： 一种单链多肽融合蛋白，其包含：非细胞毒性蛋白酶或其片段，所述蛋白酶或蛋白酶片段能够切割伤害性感觉传入的胞外融合装置的蛋白质; 能够结合伤害性感觉传入的结合位点的靶向部位，该结合位点能够经历内吞作用以掺入伤害性感觉传入内的内体; 蛋白酶切割位点，其中融合蛋白可被蛋白酶切割，其中蛋白酶切割位点位于非细胞毒性蛋白酶或其片段与靶向部位之间; 能够将位于内体内的蛋白酶或蛋白酶片段穿过内体膜并转移到伤害性感觉传入物的胞质溶胶中的易位结构域。 还描述了编码融合蛋白的核酸，其制备方法及其用途。

公开(公告)号：US20080064092A1

公开(公告)日：2008-03-13

申请号：US11853517

申请日：2007-09-11

申请人： Keith FOSTER ,  John CHADDOCK ,  Philip MARKS ,  Patrick STANCOMBE ,  Lyndsey DUROSE

发明人： Keith FOSTER ,  John CHADDOCK ,  Philip MARKS ,  Patrick STANCOMBE ,  Lyndsey DUROSE

IPC分类号： C07K14/00 ,  C12N15/62 ,  C12N15/63

CPC分类号： C12N9/50 ,  A61K38/00 ,  C07K14/33 ,  C07K2319/06 ,  C07K2319/50 ,  C12N15/62

摘要： The invention provides a single chain, polypeptide fusion protein, comprising: a non-cytotoxic protease, or a fragment thereof, which protease or protease fragment is capable of cleaving a protein of the exocytic fusion apparatus of a target cell; a Targeting Moiety that is capable of binding to a Binding Site on the target cell, which Binding Site is capable of undergoing endocytosis to be incorporated into an endosome within the target cell; a protease cleavage site at which site the fusion protein is cleavable by a protease, wherein the protease cleavage site is located between the non-cytotoxic protease or fragment thereof and the Targeting Moiety; and a translocation domain that is capable of translocating the protease or protease fragment from within an endosome, across the endosomal membrane and into the cytosol of the target cell.

摘要翻译： 本发明提供了一种单链多肽融合蛋白，其包含：非细胞毒性蛋白酶或其片段，所述蛋白酶或蛋白酶片段能够切割靶细胞的胞外融合装置的蛋白质; 能够结合目标细胞上的结合位点的靶向物质，所述结合位点能够经历内吞作用以掺入靶细胞内的内体; 蛋白酶切割位点，其中融合蛋白可被蛋白酶切割，其中蛋白酶切割位点位于非细胞毒性蛋白酶或其片段与靶向部位之间; 以及能够将位于内体内的蛋白酶或蛋白酶片段穿过体内膜并进入靶细胞的胞质溶胶的易位结构域。

公开(公告)号：US20120207735A1

公开(公告)日：2012-08-16

申请号：US13418453

申请日：2012-03-13

申请人： Keith FOSTER ,  John CHADDOCK ,  Charles PENN ,  Kei Roger AOKI ,  Joseph FRANCIS ,  Lance STEWARD

发明人： Keith FOSTER ,  John CHADDOCK ,  Charles PENN ,  Kei Roger AOKI ,  Joseph FRANCIS ,  Lance STEWARD

IPC分类号： A61K38/48 ,  A61P29/00 ,  C12N9/96 ,  C12N15/62

CPC分类号： C12N9/6432 ,  A61K38/00 ,  A61K38/22 ,  A61K38/48 ,  A61K47/48246 ,  A61K47/48338 ,  A61K47/48361 ,  A61K47/64 ,  A61K47/65 ,  A61K47/67 ,  C07K14/665 ,  C07K2319/00 ,  C07K2319/01 ,  C07K2319/21 ,  C07K2319/55 ,  C12N9/96 ,  C12N15/62 ,  C12Y304/00 ,  C12Y304/21006

摘要： The present invention is directed to non-cytotoxic protein conjugates for inhibition or reduction of exocytic fusion in a nociceptive sensory afferent cell. The protein conjugates comprise: (i) a Targeting Moiety (TM), wherein the TM is an agonist of a receptor present on a nociceptive sensory afferent cell, and wherein the receptor undergoes endocytosis to be incorporated into an endosome within the nociceptive sensory afferent cell; (ii) a non-cytotoxic protease or a fragment thereof, wherein the protease or protease fragment is capable of cleaving a protein of the exocytic fusion to apparatus of the nociceptive sensory afferent cell; and (iii) a Translocation Domain, wherein the Translocation Domain translocates the protease or protease fragment from within the endosome, across the endosomal membrane, and into the cytosol of the nociceptive sensory afferent cell wherein the Targeting Moiety is selected from the group consisting of BAM, β-endorphin, bradykinin, substance P, dynorphin and/or nociceptin.

摘要翻译： 本发明涉及用于抑制或减少伤害性感觉传入细胞中的胞外融合的非细胞毒性蛋白质缀合物。 所述蛋白质缀合物包含：（i）靶向部分（TM），其中所述TM是存在于伤害性感觉传入细胞上的受体的激动剂，并且其中所述受体经受内吞作用以掺入伤害性感觉传入细胞内的内体 ; （ii）非细胞毒性蛋白酶或其片段，其中所述蛋白酶或蛋白酶片段能够将伤感融合蛋白质切割成伤害性感觉传入细胞的装置; 易位结构域，其中易位域将内体内的蛋白酶或蛋白酶片段转移穿过内体膜，并转移到伤害性感觉传入细胞的胞质溶胶中，其中靶向部分选自BAM ，内啡肽，缓激肽，物质P，强啡肽和/或伤害感受肽。

公开(公告)号：US20110177053A1

公开(公告)日：2011-07-21

申请号：US13072865

申请日：2011-03-28

申请人： Keith FOSTER ,  John CHADDOCK ,  Charles PENN ,  Kei Roger AOKI ,  Joseph FRANCIS ,  Lance STEWARD

发明人： Keith FOSTER ,  John CHADDOCK ,  Charles PENN ,  Kei Roger AOKI ,  Joseph FRANCIS ,  Lance STEWARD

IPC分类号： A61K38/48 ,  A61P29/00 ,  C12N9/96 ,  C12N15/63

CPC分类号： C07K14/575 ,  A61K31/711 ,  A61K38/4893 ,  A61K47/64 ,  A61K47/6415 ,  C07K14/33 ,  C07K2319/00 ,  C07K2319/01 ,  C07K2319/33 ,  C12N9/6489 ,  C12N15/62 ,  C12Y304/24069

摘要： The present invention is directed to non-cytotoxic protein conjugates for inhibition or reduction of exocytic fusion in a nociceptive sensory afferent cell. The protein conjugates comprise: (i) a Targeting Moiety (TM), wherein the TM is an agonist of a receptor present on a nociceptive sensory afferent cell, and wherein the receptor undergoes endocytosis to be incorporated into an endosome within the nociceptive sensory afferent cell; (ii) a non-cytotoxic protease or a fragment thereof, wherein the protease or protease fragment is capable of cleaving a protein of the exocytic fusion apparatus of the nociceptive sensory afferent cell; and (iii) a Translocation Domain, wherein the Translocation Domain translocates the protease or protease fragment from within the endosome, across the endosomal membrane, and into the cytosol of the nociceptive sensory afferent cell. Nucleic acid sequences encoding the protein conjugates, methods of preparing same and uses thereof are also described.

摘要翻译： 本发明涉及用于抑制或减少伤害性感觉传入细胞中的胞外融合的非细胞毒性蛋白质缀合物。 所述蛋白质缀合物包含：（i）靶向部分（TM），其中所述TM是存在于伤害性感觉传入细胞上的受体的激动剂，并且其中所述受体经受内吞作用以掺入伤害性感觉传入细胞内的内体 ; （ii）非细胞毒性蛋白酶或其片段，其中所述蛋白酶或蛋白酶片段能够切割伤害性感觉传入细胞的胞外融合装置的蛋白质; 易位结构域（Translocation Domain），其中易位域将位于内体内的蛋白酶或蛋白酶片段穿过内体膜并转移到伤害性感觉传入细胞的胞质溶胶中。 还描述了编码蛋白质缀合物的核酸序列，其制备方法及其用途。

公开(公告)号：US20120156186A1

公开(公告)日：2012-06-21

申请号：US13343892

申请日：2012-01-05

申请人： Keith FOSTER ,  John CHADDOCK ,  Charles PENN ,  Kei Roger AOKI ,  Joseph FRANCIS ,  Lance STEWARD

发明人： Keith FOSTER ,  John CHADDOCK ,  Charles PENN ,  Kei Roger AOKI ,  Joseph FRANCIS ,  Lance STEWARD

IPC分类号： A61K38/48 ,  A61P21/00 ,  C12P21/00 ,  C12N9/96 ,  C12N15/57

CPC分类号： A61K38/4893 ,  A61K38/4886 ,  C07K14/575 ,  C07K2319/00 ,  C07K2319/01 ,  C12N9/52 ,  C12Y304/21072

摘要： The present invention is directed to non-cytotoxic protein conjugates for inhibition or reduction of exocytic fusion in a nociceptive sensory afferent cell. The protein conjugates comprise: (i) a galanin Targeting Moiety (TM), wherein the TM is an agonist of a receptor present on a nociceptive sensory afferent cell, and wherein the receptor undergoes endocytosis to be incorporated into an endosome within the nociceptive sensory afferent cell; (ii) a non-cytotoxic protease or a fragment thereof, wherein the protease or protease fragment is capable of cleaving a protein of the exocytic fusion apparatus of the nociceptive sensory afferent cell; and (iii) a Translocation Domain, wherein the Translocation Domain translocates the protease or protease fragment from within the endosome, across the endosomal membrane, and into the cytosol of the nociceptive sensory afferent cell. Nucleic acid sequences encoding the protein conjugates, methods of preparing same and uses thereof are is also described.

摘要翻译： 本发明涉及用于抑制或减少伤害性感觉传入细胞中的胞外融合的非细胞毒性蛋白质缀合物。 所述蛋白质缀合物包括：（i）甘丙肽靶向部分（TM），其中所述TM是存在于伤害性感觉传入细胞上的受体的激动剂，并且其中所述受体经受内吞作用以掺入伤害性感觉传入物内的内体 细胞; （ii）非细胞毒性蛋白酶或其片段，其中所述蛋白酶或蛋白酶片段能够切割伤害性感觉传入细胞的胞外融合装置的蛋白质; 易位结构域（Translocation Domain），其中易位域将位于内体内的蛋白酶或蛋白酶片段穿过内体膜并转移到伤害性感觉传入细胞的胞质溶胶中。 还描述了编码蛋白质缀合物的核酸序列，其制备方法及其用途。

公开(公告)号：US20120058098A1

公开(公告)日：2012-03-08

申请号：US13212868

申请日：2011-08-18

申请人： Keith FOSTER ,  John CHADDOCK ,  Charles PENN ,  Kei Roger AOKI ,  Joseph FRANCIS ,  Lance STEWARD

发明人： Keith FOSTER ,  John CHADDOCK ,  Charles PENN ,  Kei Roger AOKI ,  Joseph FRANCIS ,  Lance STEWARD

IPC分类号： A61K38/48 ,  C12N9/96 ,  A61P25/00 ,  C07H21/04 ,  A61P29/00

CPC分类号： C12N9/96 ,  A61K38/482 ,  A61K38/4893 ,  A61K47/48246 ,  A61K47/64 ,  C07K14/665 ,  C07K2319/00

摘要： The present invention is directed to non-cytotoxic protein conjugates for inhibition or reduction of exocytic fusion in a nociceptive sensory afferent cell. The protein conjugates comprise: (i) a dynorphin Targeting Moiety (TM), wherein the TM is an agonist of a receptor present on a nociceptive sensory afferent cell, and wherein the receptor undergoes endocytosis to be incorporated into an endosome within the nociceptive sensory afferent cell; (ii) a non-cytotoxic protease or a fragment thereof, wherein the protease or protease fragment is capable of cleaving a protein of the exocytic fusion apparatus of the nociceptive sensory afferent cell; and (iii) a Translocation Domain, wherein the Translocation Domain translocates the protease or protease fragment from within the endosome, across the endosomal membrane, and into the cytosol of the nociceptive sensory afferent cell. Nucleic acid sequences encoding the protein conjugates, methods of preparing same and uses thereof are also described.

摘要翻译： 本发明涉及用于抑制或减少伤害性感觉传入细胞中胞外融合的非细胞毒性蛋白质缀合物。 所述蛋白质缀合物包含：（i）强啡肽靶向剂量（TM），其中所述TM是感受伤害感觉传入细胞上存在的受体的激动剂，并且其中所述受体经受内吞作用以掺入伤害性感觉传入物内的内体 细胞; （ii）非细胞毒性蛋白酶或其片段，其中所述蛋白酶或蛋白酶片段能够切割伤害性感觉传入细胞的胞外融合装置的蛋白质; 易位结构域（Translocation Domain），其中易位域将位于内体内的蛋白酶或蛋白酶片段穿过内体膜并转移到伤害性感觉传入细胞的胞质溶胶中。 还描述了编码蛋白质缀合物的核酸序列，其制备方法及其用途。