公开(公告)号：US20040146963A1

公开(公告)日：2004-07-29

申请号：US10802574

申请日：2004-03-17

发明人： James J. Schmidt ,  Robert G. Stafford

IPC分类号： C07K014/33 ,  A61K039/02 ,  C12Q001/37

CPC分类号： C07K14/33 ,  C12Q1/37 ,  G01N33/542 ,  G01N33/56911 ,  G01N2333/33 ,  Y10S435/842

摘要： In this application is described substrates for high-throughput assays of clostridial neurotoxin proteolytic activities. Two types of substrates are described for use in assays for the proteolytic activities of clostridial neurotoxins: (1) modified peptides or proteins that can serve as FRET substrates and (2) modified peptides or proteins that can serve as immobilized substrates. In both types a fluorescent molecules is present in the substrate, eliminating the requirement for the addition of a fluorigenic reagent. The assays described can be readily adapted for use in automated or robotic systems.

公开(公告)号：US20030143650A1

公开(公告)日：2003-07-31

申请号：US09942024

申请日：2001-08-28

发明人： Lance E. Steward ,  Ester Fernandez-Salas ,  Kei Roger Aoki

IPC分类号： G01N033/554 ,  G01N033/569 ,  C12Q001/37 ,  C07K014/33

CPC分类号： G01N33/582

摘要： The present invention provides clostridial toxin substrates useful in assaying for the protease activity of any clostridial toxin, including botulinum toxins of all serotypes as well as tetanus toxins. A clostridial toxin substrate of the invention contains a donor fluorophore; an acceptor having an absorbance spectrum overlapping the emission spectrum of the donor fluorophore; and a clostridial toxin recognition sequence that includes a cleavage site, where the cleavage site intervenes between the donor fluorophore and the acceptor and where, under the appropriate conditions, resonance energy transfer is exhibited between the donor fluorophore and the acceptor.

摘要翻译： 本发明提供了可用于测定任何梭菌毒素（包括所有血清型的肉毒毒素）以及破伤风毒素的蛋白酶活性的梭菌毒素底物。 本发明的梭菌毒素底物含有供体荧光团; 具有与供体荧光团的发射光谱重叠的吸收光谱的受体; 和梭菌毒素识别序列，其包括切割位点，其中切割位点插入供体荧光团和受体之间，并且在适当条件下，在供体荧光团和受体之间显示共振能量转移。

公开(公告)号：US20030143651A1

公开(公告)日：2003-07-31

申请号：US09942098

申请日：2001-08-28

发明人： Lance E. Steward ,  Ester Fernandez-Salas ,  Kei Roger Aoki

IPC分类号： G01N033/554 ,  G01N033/569 ,  C12Q001/04 ,  C07K014/33

CPC分类号： C12Q1/37 ,  C07K14/001 ,  C07K14/435 ,  G01N33/56911 ,  G01N2333/33

摘要： The present invention provides clostridial toxin substrates useful in assaying for the protease activity of any clostridial toxin, including botulinum toxins of all serotypes as well as tetanus toxins. A clostridial toxin substrate of the invention contains a donor fluorophore; an acceptor having an absorbance spectrum overlapping the emission spectrum of the donor fluorophore; and a clostridial toxin recognition sequence that includes a cleavage site, where the cleavage site intervenes between the donor fluorophore and the acceptor and where, under the appropriate conditions, resonance energy transfer is exhibited between the donor fluorophore and the acceptor.

摘要翻译： 本发明提供了可用于测定任何梭菌毒素（包括所有血清型的肉毒毒素）以及破伤风毒素的蛋白酶活性的梭菌毒素底物。 本发明的梭菌毒素底物含有供体荧光团; 具有与供体荧光团的发射光谱重叠的吸收光谱的受体; 和梭菌毒素识别序列，其包括切割位点，其中切割位点插入供体荧光团和受体之间，并且在适当条件下，在供体荧光团和受体之间显示共振能量转移。

公开(公告)号：US20020197278A1

公开(公告)日：2002-12-26

申请号：US10176957

申请日：2002-06-21

申请人： SURROMED, INC.

发明人： Anthony Allison

IPC分类号： A61K039/08 ,  C07K014/33 ,  A61K039/02

CPC分类号： A61Q19/08 ,  A61K8/64 ,  A61K8/86 ,  A61K38/4893 ,  A61K47/60 ,  A61K2800/57 ,  C07K14/33 ,  C12N9/6416 ,  C12Y304/24068 ,  Y02A50/469

摘要： Modified toxins including botulinum toxin or tetanus toxin coupled to polyethylene glycol, pharmaceutical compositions of modified toxins, and methods for their use are provided. The methods include treating inappropriate muscle contraction, and treatments for cosmetic purposes.

摘要翻译： 提供了与聚乙二醇偶联的肉毒杆菌毒素或破伤风毒素，修饰毒素的药物组合物及其使用方法的修饰毒素。 这些方法包括治疗不适当的肌肉收缩和用于化妆品目的的治疗。

公开(公告)号：US20040253673A1

公开(公告)日：2004-12-16

申请号：US10728696

申请日：2003-12-05

申请人： Allergan Sales, Inc., Allergan Botox Limited

发明人： James A. Williams

IPC分类号： C07K014/33 ,  C07H021/04 ,  C12N001/21

CPC分类号： C07K14/33 ,  A61K39/00 ,  A61K2039/505 ,  C07K14/245 ,  C07K16/02 ,  C07K16/1282 ,  C07K2317/34 ,  C07K2319/00 ,  Y02A50/469

摘要： The present invention includes recombinant proteins derived from Clostridium botulinum toxins. In particular, soluble recombinant Clostridium botulinum type A, type B and type E toxin proteins are provided. Methods which allow for the isolation of recombinant proteins free of significant endotoxin contamination are provided. The soluble, endotoxin-free recombinant proteins are used as immunogens for the production of vaccines and antitoxins. These vaccines and antitoxins are useful in the treatment of humans and other animals at risk of intoxication with clostridial toxin.

摘要翻译： 本发明包括衍生自肉毒杆菌毒素的重组蛋白质。 特别地，提供可溶性重组肉毒杆菌A型，B型和E型毒素蛋白。 提供了允许分离没有显着内毒素污染的重组蛋白质的方法。 可溶性，不含内毒素的重组蛋白被用作生产疫苗和抗毒素的免疫原。 这些疫苗和抗毒素可用于治疗患有梭菌毒素中毒风险的人类和其他动物。

公开(公告)号：US20040126827A1

公开(公告)日：2004-07-01

申请号：US10766283

申请日：2004-01-27

申请人： University of Massachusetts, a Massachusetts corporation

发明人： Bal Ram Singh ,  Zhong Zhang

IPC分类号： C07K014/33 ,  G01N033/554 ,  G01N033/569

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

摘要： The invention features a polypeptide complex synthesized by bacteria of the genus Clostridia that contains the serotype E botulinum neurotoxin and five neurotoxin associated polypeptides having molecular weights of about 118, 80, 65, 40, and 18 kDa. respectively. The complex is useful in the treatment of diseases or conditions that are caused by excessive release of acetylcholine from presynaptic nerve terminals.

摘要翻译： 本发明的特征在于由梭菌属细菌合成的多肽复合物，其包含血清型E肉毒杆菌神经毒素和五种具有约118,80,65,40和18kDa分子量的神经毒素相关多肽。 分别。 该复合物可用于治疗由突触前神经末梢过度释放乙酰胆碱引起的疾病或病症。

公开(公告)号：US20030077685A1

公开(公告)日：2003-04-24

申请号：US09962360

申请日：2001-09-25

发明人： James J. Schmidt ,  Robert G. Stafford

IPC分类号： C12Q001/37 ,  C12Q001/04 ,  C07K014/33

CPC分类号： C07K14/33 ,  C12Q1/37 ,  G01N33/542 ,  G01N33/56911 ,  G01N2333/33 ,  Y10S435/842

摘要： In this application is described substrates for high-throughput assays of clostridial neurotoxin proteolytic activities. Two types of substrates are described for use in assays for the proteolytic activities of clostridial neurotoxins: (1) modified peptides or proteins that can serve as FRET substrates and (2) modified peptides or proteins that can serve as immobilized substrates. In both types a fluorescent molecules is present in the substrate, eliminating the requirement for the addition of a fluorigenic reagent. The assays described can be readily adapted for use in automated or robotic systems.

摘要翻译： 在本申请中描述了用于梭菌神经毒素蛋白水解活性的高通量测定的底物。 描述了两种类型的底物用于梭菌神经毒素的蛋白水解活性的测定：（1）可用作FRET底物的修饰肽或蛋白质，和（2）可用作固定化底物的修饰肽或蛋白质。 在两种类型中，荧光分子存在于底物中，消除了添加荧光试剂的要求。 所描述的测定法可以容易地适用于自动化或机器人系统。

公开(公告)号：US20040220386A1

公开(公告)日：2004-11-04

申请号：US10757077

申请日：2004-01-14

发明人： Lance E. Steward ,  Ester Fernandez-Salas ,  Todd M. Herrington ,  Kei Roger Aoki

IPC分类号： C07K014/33 ,  A61K039/08

CPC分类号： C07K14/33 ,  A61K38/00 ,  C07K2319/01

摘要： Natural and modified neurotoxins and isolated neurotoxin compositions are described. The neurotoxins may include one or more structural modifications, wherein the structural modification(s) alters the biological persistence, such as the biological half-life and/or a biological activity of the modified neurotoxin relative to an identical neurotoxin without the structural modification(s). In one embodiment, methods of making the modified neurotoxin include using recombinant techniques. In some embodiments, methods of using the modified neurotoxin to treat conditions include treating various disorders, neuromuscular ailments and pain.

摘要翻译： 描述了天然和修饰的神经毒素和分离的神经毒素组合物。 神经毒素可以包括一种或多种结构修饰，其中结构修饰改变了相对于相同的神经毒素而没有结构修饰的生物学持久性，例如修饰的神经毒素的生物半衰期和/或生物活性 ）。 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。 在一些实施方案中，使用经修饰的神经毒素治疗病症的方法包括治疗各种疾病，神经肌肉疾病和疼痛。

公开(公告)号：US20040072270A1

公开(公告)日：2004-04-15

申请号：US10261161

申请日：2002-09-27

发明人： Ester Fernandez-Salas ,  Lance E. Steward ,  Kei Roger Aoki

IPC分类号： G01N033/554 ,  G01N033/569 ,  C12Q001/37 ,  C07K014/33

CPC分类号： C07K14/005 ,  C07K14/33 ,  C07K14/4702 ,  C07K14/4725 ,  C12N2710/16622 ,  C12N2740/16122 ,  C12Q1/37

摘要： The present invention provides a method of determining clostridial toxin activity by (a) contacting with a sample a cell containing a clostridial toxin substrate that includes a donor fluorophore; an acceptor having an absorbance spectrum overlapping the emission spectrum of the donor fluorophore; and a clostridial toxin recognition sequence containing a cleavage site that intervenes between the donor fluorophore and the acceptor, where resonance energy transfer is exhibited between the donor fluorophore and the acceptor under the appropriate conditions; (b) exciting the donor fluorophore; and (c) determining resonance energy transfer of the contacted cell relative to a control cell, where a difference in resonance energy transfer of the contacted cell as compared to the control cell is indicative of clostridial toxin activity.

摘要翻译： 本发明提供了一种通过（a）使样品与包含供体荧光团的梭菌毒素底物的细胞接触来测定梭菌毒素活性的方法; 具有与供体荧光团的发射光谱重叠的吸收光谱的受体; 以及包含介入供体荧光团和受体之间的切割位点的梭菌毒素识别序列，其中在适当条件下在供体荧光团和受体之间显示共振能量转移; （b）激发供体荧光团; 和（c）确定接触细胞相对于对照细胞的共振能量转移，其中与对照细胞相比，所接触的细胞的共振能量转移的差异指示梭菌毒素活性。

公开(公告)号：US20040028705A1

公开(公告)日：2004-02-12

申请号：US10463957

申请日：2003-06-17

发明人： Jimmy D. Ballard ,  Lea M. Spyres

IPC分类号： A61K039/08 ,  C12N001/21 ,  C07K014/33

CPC分类号： G01N33/56911 ,  A61K38/00 ,  A61K39/00 ,  C07K14/33

摘要： An active or passive vaccine utilizing purified non-toxic mutant TcdB toxins from Clostridium difficile for humans and animals against infections caused by C. difficile and/or C. sordellii. Persons most potentially affected by C. difficile infections include hospitalized patients, infants, and elderly persons. The TcdB toxin mutant of the vaccine preferably lacks the toxicity of a native C. difficile TcdB toxin. A serum comprising antibodies raised to the TcdB toxin mutant is also available for treating humans or animals against C. difficile infections. The serum may be used in a method for conferring passive immunity against C. difficile. Antibodies to the TcdB toxin mutant may be used in diagnostic tests or in treatments to clear TcdB toxin from bodily fluids. The mutant TcdB toxin may be produced by recombinant methods using cDNA encoding the toxin, the cDNA contained for example in a plasmid or host cell.

摘要翻译： 使用来自艰难梭菌（Clostridium difficile）的纯化的无毒突变型TcdB毒素用于人和动物针对艰难梭菌和/或C.sordellii引起的感染的活性或被动疫苗。 最可能受艰难梭菌感染影响的人群包括住院患者，婴儿和老年人。 疫苗的TcdB毒素突变体优选缺乏天然艰难梭菌TcdB毒素的毒性。 包含提高至TcdB毒素突变体的抗体的血清也可用于治疗人或动物针对艰难梭菌感染。 该血清可用于赋予难治性艰难梭菌被动免疫的方法。 TcdB毒素突变体的抗体可用于诊断测试或用于从体液中清除TcdB毒素的治疗。 突变型TcdB毒素可以使用编码毒素的cDNA的重组方法产生，cDNA包含在例如质粒或宿主细胞中。