公开(公告)号：US08309686B2

公开(公告)日：2012-11-13

申请号：US12212774

申请日：2008-09-18

申请人： Wei-Jen Lin ,  Kei Roger Aoki ,  Lance E. Steward

发明人： Wei-Jen Lin ,  Kei Roger Aoki ,  Lance E. Steward

IPC分类号： C12P21/06

CPC分类号： C07K14/33 ,  A61K38/00 ,  Y02A50/469

摘要： Modified neurotoxins that contain protease cleavage sites susceptible uniquely to proteases present in certain tissues are described. The toxins can be selectively activated by proteases in muscle or selectively inactivated by proteases in blood.

摘要翻译： 描述了含有蛋白酶切割位点的修饰的神经毒素，其特异性地存在于某些组织中存在的蛋白酶。 毒素可以被肌肉中的蛋白酶选择性活化或被血液中的蛋白酶选择性地灭活。

公开(公告)号：US07723480B2

公开(公告)日：2010-05-25

申请号：US12055454

申请日：2008-03-26

申请人： Lance E. Steward ,  Todd M. Herrington ,  Kei Roger Aoki

发明人： Lance E. Steward ,  Todd M. Herrington ,  Kei Roger Aoki

IPC分类号： C07K14/33

CPC分类号： C07K14/33 ,  A61K38/00 ,  Y10S530/825

摘要： Modified neurotoxin comprising neurotoxin including structural modification, wherein the structural modification alters the biological persistence, preferably the biological half-life, of the modified neurotoxin relative to an identical neurotoxin without the structural modification. The structural modification includes addition or deletion of a leucine-based motif or parts thereof. In one embodiment, methods of making the modified neurotoxin include using recombinant techniques. In another embodiment, methods of using the modified neurotoxin to treat biological disorders include treating autonomic disorders, neuromuscular disorders or pains.

公开(公告)号：US07705124B2

公开(公告)日：2010-04-27

申请号：US12053998

申请日：2008-03-24

申请人： Lance E. Steward ,  Todd M. Herrington ,  Kei Roger Aoki

发明人： Lance E. Steward ,  Todd M. Herrington ,  Kei Roger Aoki

IPC分类号： C07K14/33

CPC分类号： C07K14/33 ,  A61K38/00 ,  Y10S530/825

摘要： Modified neurotoxin comprising neurotoxin including structural modification, wherein the structural modification alters the biological persistence, preferably the biological half-life, of the modified neurotoxin relative to an identical neurotoxin without the structural modification. The structural modification includes addition or deletion of a leucine-based motif or parts thereof. In one embodiment, methods of making the modified neurotoxin include using recombinant techniques. In another embodiment, methods of using the modified neurotoxin to treat biological disorders include treating autonomic disorders, neuromuscular disorders or pains.

公开(公告)号：US07393925B2

公开(公告)日：2008-07-01

申请号：US11039268

申请日：2005-01-19

申请人： Lance E. Steward ,  Todd M. Herrington ,  Kei Roger Aoki

发明人： Lance E. Steward ,  Todd M. Herrington ,  Kei Roger Aoki

IPC分类号： C07K14/33

CPC分类号： C07K14/33 ,  A61K38/00 ,  Y10S530/825

摘要： Modified neurotoxin comprising neurotoxin including structural modification, wherein the structural modification alters the biological persistence, preferably the biological half-life, of the modified neurotoxin relative to an identical neurotoxin without the structural modification. The structural modification includes addition or deletion of a leucine-based motif or parts thereof. In one embodiment, methods of making the modified neurotoxin include using recombinant techniques. In another embodiment, methods of using the modified neurotoxin to treat biological disorders include treating autonomic disorders, neuromuscular disorders or pains.

摘要翻译： 包含神经毒素的改性神经毒素包括结构修饰，其中所述结构修饰改变了修饰神经毒素相对于相同神经毒素的生物学持续性，优选生物半衰期而没有结构修饰。 结构修饰包括加入或缺失基于亮氨酸的基序或其部分。 在一个实施方案中，制备修饰的神经毒素的方法包括使用重组技术。 在另一个实施方案中，使用经修饰的神经毒素治疗生物学障碍的方法包括治疗自主神经障碍，神经肌肉疾病或疼痛。

公开(公告)号：US06991789B2

公开(公告)日：2006-01-31

申请号：US10880192

申请日：2004-06-29

申请人： Shengwen Li ,  Kei Roger Aoki

发明人： Shengwen Li ,  Kei Roger Aoki

IPC分类号： A61K38/48

CPC分类号： A61K9/0019 ,  A61K38/4886

摘要： The present invention provides for methods of modulating the degradation rate of a toxin in a cell, thereby modulating the half-life of the toxin. Particularly, the invention features methods of modulating the degradation rate of a toxin by modulating fusion between a lysosome and an endosome that carries the toxin in the cell.

公开(公告)号：US08399401B2

公开(公告)日：2013-03-19

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： C07K14/00 ,  C07H21/02

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：US20120230975A1

公开(公告)日：2012-09-13

申请号：US13419381

申请日：2012-03-13

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 ,  C12N9/52 ,  A61P25/00 ,  C12N15/63 ,  A61P29/00 ,  C12N9/50 ,  C12N15/62

CPC分类号： C12Y304/24069 ,  A61K38/00 ,  A61K38/1709 ,  A61K38/482 ,  A61K38/4886 ,  A61K38/4893 ,  A61K47/6415 ,  A61K47/65 ,  C07K14/665 ,  C07K2319/055 ,  C07K2319/06 ,  C07K2319/50 ,  C12N9/52 ,  C12N15/62

摘要： A single chain, polypeptide fusion protein, comprising: a non-cytotoxic protease, or a fragment thereof, which protease or protease fragment can cleave a protein of the exocytic fusion apparatus of a nociceptive sensory afferent; a Targeting Moiety that can bind to a Binding Site on the nociceptive sensory afferent, which Binding Site can undergo 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, which is located between the non-cytotoxic protease and the Targeting Moiety; and a translocation domain that can translocate the protease or protease fragment from within an endosome, across the endosomal membrane and into the cytosol of the nociceptive sensory afferent; wherein the Targeting Moiety is BAM, β-endorphin, bradykinin, substance P, dynorphin and/or nociceptin. Nucleic acid sequences 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可以切割伤害性感觉传入的胞外融合装置的蛋白质; 可以结合伤害性感觉传入物上的结合位点的靶向物质，该结合位点可以进行内吞作用以掺入伤害性感觉传入体内的内体; 蛋白酶切割位点，位于融合蛋白可被位于非细胞毒性蛋白酶和靶向部位之间的蛋白酶切割; 以及易位区域，其可以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; 其中靶向部分是BAM，β-内啡肽，缓激肽，物质P，强啡肽和/或伤害感受肽。 还描述了编码融合蛋白的核酸序列，其制备方法及其用途。

公开(公告)号：US08067200B2

公开(公告)日：2011-11-29

申请号：US11792210

申请日：2005-12-01

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： C12P21/06 ,  C17H17/00 ,  C07K14/00

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; 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.

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; 以及易位区域，其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。 还描述了编码多肽融合蛋白的核酸序列，其制备方法及其用途。

公开(公告)号：US20100203559A1

公开(公告)日：2010-08-12

申请号：US12723474

申请日：2010-03-12

申请人： Fernandez-Salas Ester ,  Joanne Wang ,  Patton Garay ,  Lina S. Wong ,  D. Dianne Hodges ,  Kei Roger Aoki

发明人： Fernandez-Salas Ester ,  Joanne Wang ,  Patton Garay ,  Lina S. Wong ,  D. Dianne Hodges ,  Kei Roger Aoki

IPC分类号： G01N33/53 ,  C07K7/08 ,  C07K16/18 ,  C07K7/06

CPC分类号： C07K16/1282 ,  C07K2317/34 ,  C07K2317/77 ,  C12Q1/37 ,  G01N33/56911 ,  G01N2333/33 ,  G01N2333/705 ,  G01N2333/952

摘要： The present specification discloses SNAP-25 compositions, methods of making α-SNAP-25 antibodies that bind an epitope comprising a carboxyl-terminus at the P1 residue from the BoNT/A cleavage site scissile bond from a SNAP-25 cleavage product, α-SNAP-25 antibodies that bind an epitope comprising a carboxyl-terminus at the P1 residue from the BoNT/A cleavage site scissile bond from a SNAP-25 cleavage product, methods of detecting BoNT/A activity, and methods of detecting neutralizing α-BoNT/A antibodies.

摘要翻译： 本说明书公开了SNAP-25组合物，制备α-SNAP-25抗体的方法，所述α-SNAP-25抗体结合来自SNAP-25裂解产物的BoNT / A切割位点剪切键的P1残基处的包含羧基末端的表位， SNAP-25抗体结合来自SNAP-25切割产物的BoNT / A切割位点剪切键的P1残基处的羧基末端的表位，检测BoNT / A活性的方法和检测中和α-BoNT的方法 / A抗体。

公开(公告)号：US07674601B2

公开(公告)日：2010-03-09

申请号：US12125327

申请日：2008-05-22

申请人： Dudley J. Williams ,  Marcella Gilmore ,  Lance Steward ,  Marc Verhagen ,  Kei Roger Aoki

发明人： Dudley J. Williams ,  Marcella Gilmore ,  Lance Steward ,  Marc Verhagen ,  Kei Roger Aoki

IPC分类号： C12Q1/37 ,  C12N11/16

CPC分类号： G01N33/542 ,  C12N9/52 ,  C12Q1/37 ,  G01N2333/33

摘要： The present invention provides a method of determining the presence or activity of a clostridial toxin by (a) treating with a sample, under conditions suitable for clostridial toxin protease activity, a clostridial toxin substrate which includes a fluorophore; a bulking group; and a clostridial toxin recognition sequence containing a cleavage site that intervenes between the fluorophore and the bulking group; (b) exciting the fluorophore with plane polarized light; and (c) determining fluorescence polarization of the treated substrate relative to a control substrate, where a change in fluorescence polarization of the treated substrate as compared to fluorescence polarization of the control substrate is indicative of the presence or activity of the clostridial toxin.

摘要翻译： 本发明提供一种通过（a）在适合于梭菌毒素蛋白酶活性的条件下用样品处理包括荧光团的梭菌毒素底物来确定梭菌毒素的存在或活性的方法; 一个胖胖的小组 和含有插入在荧光团和膨化基团之间的切割位点的梭菌毒素识别序列; （b）用平面偏振光激发荧光团; 和（c）确定经处理的底物相对于对照底物的荧光偏振，其中与对照底物的荧光偏振相比，经处理的底物的荧光偏振的变化指示梭菌毒素的存在或活性。