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1. 发明公开

EP2245455A1 METHOD FOR DISTINGUISHING SECRETORY GRANULES OF DIFFERENT AGES 有权
标题翻译： VERFAHREN ZUR UNTERSCHEIDUNG SEKRETORISCHER GRANULA UNTERSCHIEDLICHEN ALTERS
公开(公告)号：EP2245455A1
公开(公告)日：2010-11-03
申请号：EP09702668.6
申请日：2009-01-16
申请人： Technische Universität Dresden Medizinische Fakultät Carl Gustav Carus
发明人： SOLIMENA, Michele , IVANOVA, Anna
IPC分类号： G01N33/50 , A01K67/027
CPC分类号： G01N33/5076 , A01K67/0275 , A01K2217/07 , A01K2227/105 , A01K2267/0393 , C07K2319/036 , C12N15/8509 , G01N33/5005 , G01N33/74
摘要： The present invention relates to an in vitro method of detecting secretory granules (SGs) of different ages in a cell by differentially labelling said SGs according to their age comprising (a) contacting a cell capable of forming SGs and expressing 1. a (poly)peptide specific for SGs, wherein said (poly)peptide has a binding site common for at least three different substances A, B and C covalently binding thereto or 2. a fusion protein comprising (i) a (poly)peptide specific for SGs and (ii) a (poly)peptide having a binding site common for at least three different substances A, B and C covalently binding thereto, wherein said substances are capable of penetrating the cell membrane and wherein at least substances A and C are detectable by different means with a substance A targeting said binding site (b) saturating said binding sites by contacting the cell with a substance B which blocks the binding sites which were not bound by substance A, (c) allowing that unbound substance B is removed from the cell or removing unbound substance B from the cell, (d) contacting the cell with a substance C targeting said binding site on the (poly)peptide or the fusion protein becoming available for binding after step (b) and (e) detecting for the presence of both detectable substances A and C, wherein the simultaneous detection of both substances A and C is indicative of the presence of two populations of SGs having a different age. Further, the present invention relates to an in vitro method of investigating in a cell the effect of a stimulus on secretory granules (SGs) differentially labelled according to their age.
摘要翻译：本发明涉及通过根据其年龄差异标记所述SGs来检测细胞中不同年龄的分泌颗粒（SG）的体外方法，其包括（a）使能够形成SG的细胞接触并表达1.a（聚）对于SGs特异性肽，其中所述（多）肽具有与至少三种与其共价结合的至少三种不同物质A，B和C的结合位点，或2.融合蛋白，其包含（i）特异于SG的（多）肽和（ ii）具有与至少三种与其共价结合的至少三种不同物质A，B和C的结合位点的（多）肽，其中所述物质能够穿透细胞膜，并且其中至少物质A和C可通过不同的方式检测与靶向所述结合位点的物质A（b）通过使细胞与阻断未被物质A结合的结合位点的物质B接触而饱和所述结合位点，（c）允许未结合物质B被去除从细胞中移出或从细胞中除去未结合的物质B，（d）使细胞与在步骤（b）和（e）之后可用于结合的（多）肽或融合蛋白上靶向所述结合位点的物质C接触，检测两种可检测物质A和C的存在，其中同时检测物质A和C两者指示存在两个具有不同年龄的SG群体。此外，本发明涉及一种在细胞中研究刺激对根据其年龄差异标记的分泌颗粒（SG）的影响的体外方法。

2. 发明公开

EP1632245A1 ICA512 couples insulin secretion and gene expression in Beta-cells 审中-公开
标题翻译： ICA512 koppelt Insulinsekeretion und Genexpression in Beta-Zellen
公开(公告)号：EP1632245A1
公开(公告)日：2006-03-08
申请号：EP04020912.4
申请日：2004-09-02
申请人： Technische Universität Dresden Medizinische Fakultät Carl Gustav Carus
发明人： Trajkovski, Mirko , Mziaut, Hassan , Solimena, Michele
IPC分类号： A61K38/17 , A61P3/10
CPC分类号： A61K38/1709
摘要： The present invention relates to a method for stimulating expression of peptide hormones in peptide-hormone secreting endocrine cells or neurons comprising the step of promoting in said cells or neurons the presence or activity (aa) of (i) ICA512; or (ii) a derivative thereof having ICA512 function; or (iii) a fragment of ICA512 that may be cleaved by µ-calpain giving rise to a C-terminal fragment of ICA512 wherein said C-terminal fragment has the capability of being targeted to the nucleus; or (iv) a derivative of said fragment of (iii) that may be cleaved by µ-calpain giving rise to a derivative of said C-terminal fragment of ICA512 wherein said derivative of said C-terminal fragment has the capability of being targeted to the nucleus; or (v) a fragment or derivative of ICA512 that may be cleaved by µ-calpain giving rise to a C-terminal fragment of ICA512 or derivative thereof wherein said C-terminal fragment or derivative thereof has the capability of interacting with a PIAS protein in said cells or neurons; or (vi) a fragment or derivative of ICA512 that may be cleaved by µ-calpain giving rise to a C-terminal fragment of ICA512 or derivative thereof wherein said C-terminal fragment or derivative thereof has the capability of enhancing the nuclear levels, or tyrosine phosphorylation, or DNA binding activity of STATs in said cells or neurons; or (vii) a pro-form of any one of (i) to (vi); and (ab) optionally of µ-calpain or a fragment or derivative thereof having µ-calpain function; or (b) of a C-terminal fragment of ICA512 or a derivative thereof which has the capability of being targeted to the nucleus or which has the capability of interacting with a PIAS protein or of enhancing the nuclear levels, or tyrosine phosphorylation, or DNA binding activity of STATs in said cells or neurons. Additionally, the present invention relates to a method of promoting cell proliferation of peptide-hormone secreting endocrine cells or neurons comprising the step of promoting in said cells or neurons the presence or activity (aa) of (i) ICA512; or (ii) a derivative thereof having ICA512 function; or (iii) a fragment of ICA512 that may be cleaved by µ-calpain giving rise to a C-terminal fragment of ICA512 wherein said C-terminal fragment has the capability of being targeted to the nucleus; or (iv) a derivative of said fragment of (iii) that may be cleaved by µ-calpain giving rise to a derivative of said C-terminal fragment of ICA512 wherein said derivative of said C-terminal fragment has the capability of being targeted to the nucleus; or (v) a fragment or derivative of ICA512 that may be cleaved by µ-calpain giving rise to a C-terminal fragment of ICA512 or derivative thereof wherein said C-terminal fragment or derivative thereof has the capability of interacting with a PIAS protein in said cells or neurons; or (vi) a fragment or derivative of ICA512 that may be cleaved by µ-calpain giving rise to a C-terminal fragment of ICA512 or derivative thereof wherein said C-terminal fragment or derivative thereof has the capability of enhancing the nuclear levels, or tyrosine phosphorylation, or DNA binding activity of STATs in said cells or neurons; or (vii) a pro-form of any one of (i) to (vi); and (ab) optionally of µ-calpain or a fragment or derivative thereof having µ-calpain function; or (b) of a C-terminal fragment of ICA512 or a derivative thereof which has the capability of being targeted to the nucleus or which has the capability of interacting with a PIAS protein or of enhancing the nuclear levels, or tyrosine phosphorylation, or DNA binding activity of STATs in said cells or neurons. It is preferred in accordance with the invention that said endocrine cells are β-cells and that said peptide hormone is insulin.
摘要翻译：本发明涉及刺激肽激素分泌内分泌细胞或神经元中肽激素表达的方法，其包括在所述细胞或神经元中促进（i）ICA512的存在或活性（aa）的步骤; 或（ii）具有ICA512功能的衍生物; 或（iii）ICA512的片段，其可被μ-钙蛋白酶切割，产生ICA512的C-末端片段，其中所述C-末端片段具有靶向细胞核的能力; 或（iv）可被μ-钙蛋白酶切割的（iii）的所述片段的衍生物，产生ICA512的所述C-末端片段的衍生物，其中所述C-末端片段的所述衍生物具有被靶向于细胞核或（v）ICA512的片段或衍生物，其可被μ钙蛋白酶切割，产生ICA512或其衍生物的C末端片段，其中所述C-末端片段或其衍生物具有与PIAS蛋白相互作用的能力说细胞或神经元; 或（vi）ICA512的片段或衍生物，其可被μ钙蛋白酶切割，产生ICA512或其衍生物的C末端片段，其中所述C-末端片段或其衍生物具有增强核水平的能力，或酪氨酸磷酸化或STAT在所述细胞或神经元中的DNA结合活性; 或（vii）（i）至（vi）任何一种的形式; 和（ab）任选的具有μ钙蛋白酶功能的μ-钙蛋白酶或其片段或衍生物; 或（b）ICA512的C-末端片段或其衍生物，其具有靶向细胞核的能力或具有与PIAS蛋白相互作用或增强核水平或酪氨酸磷酸化或DNA的能力 STAT在所述细胞或神经元中的结合活性。另外，本发明涉及促进分泌肽激素的内分泌细胞或神经元的细胞增殖的方法，其包括在所述细胞或神经元中促进（i）ICA512的存在或活性（aa）的步骤; 或（ii）具有ICA512功能的衍生物; 或（iii）ICA512的片段，其可被μ-钙蛋白酶切割，产生ICA512的C-末端片段，其中所述C-末端片段具有靶向细胞核的能力; 或（iv）可被μ-钙蛋白酶切割的（iii）的所述片段的衍生物，产生ICA512的所述C-末端片段的衍生物，其中所述C-末端片段的所述衍生物具有被靶向于细胞核或（v）ICA512的片段或衍生物，其可被μ钙蛋白酶切割，产生ICA512或其衍生物的C末端片段，其中所述C-末端片段或其衍生物具有与PIAS蛋白相互作用的能力说细胞或神经元; 或（vi）ICA512的片段或衍生物，其可被μ钙蛋白酶切割，产生ICA512或其衍生物的C末端片段，其中所述C-末端片段或其衍生物具有增强核水平的能力，或酪氨酸磷酸化或STAT在所述细胞或神经元中的DNA结合活性; 或（vii）（i）至（vi）任何一种的形式; 和（ab）任选的具有μ钙蛋白酶功能的μ-钙蛋白酶或其片段或衍生物; 或（b）ICA512的C-末端片段或其衍生物，其具有靶向细胞核的能力或具有与PIAS蛋白相互作用或增强核水平或酪氨酸磷酸化或DNA的能力 STAT在所述细胞或神经元中的结合活性。根据本发明，优选的是所述内分泌细胞是2细胞，而所述肽激素是胰岛素。

3. 发明公开

EP2093564A1 Method for distinguishing secretory granules of different ages 审中-公开
标题翻译： Verfahren zur Unterscheidung von Sekretgranula verschiedener Altersgruppen
公开(公告)号：EP2093564A1
公开(公告)日：2009-08-26
申请号：EP08003393.9
申请日：2008-02-25
申请人： Technische Universität Dresden Medizinische Fakultät Carl Gustav Carus
发明人： Solimena, Michele , Ivanova, Anna
IPC分类号： G01N33/50
CPC分类号： G01N33/507 , G01N33/5076
摘要： The present invention relates to an in vitro method of detecting secretory granules (SGs) of different ages in a cell by differentially labelling said SGs according to their age comprising (a) contacting a cell capable of forming SGs and expressing 1. a (poly)peptide specific for SGs, wherein said (poly)peptide has a binding site common for at least three different substances A, B and C covalently binding thereto or 2. a fusion protein comprising (i) a (poly)peptide specific for SGs and (ii) a (poly)peptide having a binding site common for at least three different substances A, B and C covalently binding thereto, wherein said substances are capable of penetrating the cell membrane and wherein at least substances A and C are detectable by different means with a substance A targeting said binding site (b) saturating said binding sites by contacting the cell with a substance B which blocks the binding sites which were not bound by substance A, (c) allowing that unbound substance B is removed from the cell or removing unbound substance B from the cell, (d) contacting the cell with a substance C targeting said binding site on the (poly)peptide or the fusion protein becoming available for binding after step (b) and (e) detecting for the presence of both detectable substances A and C, wherein the simultaneous detection of both substances A and C is indicative of the presence of two populations of SGs having a different age. Further, the present invention relates to an in vitro method of investigating in a cell the effect of a stimulus on secretory granules (SGs) differentially labelled according to their age.
摘要翻译：本发明涉及通过根据其年龄差异标记所述SGs来检测细胞中不同年龄的分泌颗粒（SG）的体外方法，其包括（a）使能够形成SG的细胞接触并表达1.a（聚）对于SGs特异性肽，其中所述（多）肽具有与至少三种与其共价结合的至少三种不同物质A，B和C的结合位点，或2.融合蛋白，其包含（i）特异于SG的（多）肽和（ ii）具有与至少三种与其共价结合的至少三种不同物质A，B和C的结合位点的（多）肽，其中所述物质能够穿透细胞膜，并且其中至少物质A和C可通过不同的方式检测与靶向所述结合位点的物质A（b）通过使细胞与阻断未被物质A结合的结合位点的物质B接触而饱和所述结合位点，（c）允许未结合的物质B被去除从细胞中移出或从细胞中除去未结合的物质B，（d）使细胞与在步骤（b）和（e）之后可用于结合的（多）肽或融合蛋白上靶向所述结合位点的物质C接触，检测两种可检测物质A和C的存在，其中同时检测物质A和C两者指示存在两个具有不同年龄的SG群体。此外，本发明涉及一种在细胞中研究刺激对根据其年龄差异标记的分泌颗粒（SG）的影响的体外方法。

4. 发明公开

EP1784207A1 ICA512 COUPLES INSULIN SECRETIN AND GENE EXPRESSION IN BETA CELLS 审中-公开
标题翻译： ICA512停靠胰岛素分泌和基因表达的β细胞
公开(公告)号：EP1784207A1
公开(公告)日：2007-05-16
申请号：EP05791045.7
申请日：2005-09-02
申请人： Technische Universität Dresden Medizinische Fakultät Carl Gustav Carus
发明人： SOLIMENA, Michele , MZIAUT, Hassan , TRAJKOVSKI, Mirko
IPC分类号： A61K38/17 , A61P3/10
CPC分类号： A61K38/1709
摘要： The present invention relates to a method for stimulating expression of peptide hormones in peptide-hormone secreting endocrine cells or neurons comprising the step of promoting in said cells the presence or activity of ICA512, a derivative or a fragment thereof that may be cleaved by mu-calpain giving rise to a C-terminal fragment of ICA512 having the capability to be targeted to the nucleus. It is preferred that said endocrine cells are betacells and that said peptide hormone is insulin.

5. 发明授权

EP2245455B1 METHOD FOR DISTINGUISHING SECRETORY GRANULES OF DIFFERENT AGES 有权
标题翻译：方法DISTINCTION分泌颗粒剂不同年龄
公开(公告)号：EP2245455B1
公开(公告)日：2014-07-09
申请号：EP09702668.6
申请日：2009-01-16
申请人： Technische Universität Dresden Medizinische Fakultät Carl Gustav Carus
发明人： SOLIMENA, Michele , IVANOVA, Anna
IPC分类号： G01N33/50 , A01K67/027
CPC分类号： G01N33/5076 , A01K67/0275 , A01K2217/07 , A01K2227/105 , A01K2267/0393 , C07K2319/036 , C12N15/8509 , G01N33/5005 , G01N33/74

6. 发明公开

EP1987837A1 Pharmaceutical composition for increasing insulin secretion from pancreatic ß-cells 审中-公开
标题翻译：药剂师Zusammensetzung zurErhöhungder Insulinsekretion aus Pankreas-ß-Zellen
公开(公告)号：EP1987837A1
公开(公告)日：2008-11-05
申请号：EP07008880.2
申请日：2007-05-02
申请人： Technische Universität Dresden Medizinische Fakultät Carl Gustav Carus
发明人： Solimena, Michele , Trajkovski, Mirko , Mziaut, Hassan , Schubert, Sandra
IPC分类号： A61K38/17 , G01N33/50 , A61K31/66 , A61P3/10
CPC分类号： A61K38/1709 , A61K31/661 , G01N33/566 , G01N2500/04
摘要： The present invention relates to a pharmaceutical composition comprising (a) ICA512, ICA512-TMF or ICA512-CCF, each devoid of the C-terminus or PHOGRIN devoid of the C-terminus or of a mimetic of the C-terminus of ICA512, ICA512-TMF, ICA512-CCF or PHOGRIN; (b) a compound promoting the presence or activity of a protease that cleaves the C-terminus of ICA512, ICA512-TMF or ICA512-CCF or PHOGRIN; (c) a compound promoting the presence or activity of a serine/threonine kinase which phosphorylates β2-syntrophin on serine residue 95; (d) a compound interfering with the presence or activity of a serine/threonine phosphatase which dephosphorylates β2-syntrophin on serine residue 95; (e) a compound promoting the presence or activity of a serine/threonine phosphatase which dephosphorylates β2-syntrophin on serine residue 110; or (f) a compound interfering with the presence or activity of a serine/threonine kinase which phosphorylates β2-syntrophin on serine residue 110.
摘要翻译：本发明涉及一种药物组合物，其包含（a）ICA512，ICA512-TMF或ICA512-CCF，其各自没有C末端或没有C末端的ICAO512，ICA512的C-末端的模拟物 -TMF，ICA512-CCF或PHOGRIN; （b）促进切割ICA512，ICA512-TMF或ICA512-CCF或PHOGRIN的C末端的蛋白酶的存在或活性的化合物; （c）促进丝氨酸/苏氨酸激酶的存在或活性的化合物，其使丝氨酸残基95上的2-合成蛋白磷酸化; （d）干扰丝氨酸/苏氨酸磷酸酶的存在或活性的化合物，其将丝氨酸残基95上的2-合成蛋白去磷酸化; （e）促进丝氨酸/苏氨酸磷酸酶的存在或活性的化合物，其将丝氨酸残基110上的2-合成蛋白脱磷酸化; 或（f）干扰丝氨酸/苏氨酸激酶的存在或活性的化合物，其在丝氨酸残基110上磷酸化2-合成蛋白。

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