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1. 发明授权

US09695422B2 Furin-knockdown bi-functional RNA 有权
公开(公告)号：US09695422B2
公开(公告)日：2017-07-04
申请号：US14844912
申请日：2015-09-03
申请人： GRADALIS, INC.
发明人： John J. Nemunaitis , Neil Senzer , Phillip B. Maples , Donald Rao
IPC分类号： C12N15/00 , C07H21/04 , C07K1/00 , C12N15/113 , A61K38/21 , C12N15/85 , C07K14/535 , A61K38/19 , A61K48/00
CPC分类号： C12N15/1137 , A61K38/193 , A61K38/217 , A61K48/005 , C07K14/535 , C12N15/85 , C12N2310/14 , C12N2310/141 , C12N2310/3519 , C12N2310/531 , C12N2320/31 , C12N2320/35 , C12N2330/51 , C12N2830/20 , C12N2840/107 , A61K2300/00
摘要： Compositions and methods to attenuate the immunosuppressive activity of TGF-β through the use of bi-functional shRNAs is described herein. The bi-functional shRNAs of the present invention knocks down the expression of furin in cancer cells to augment tumor antigen expression, presentation, and processing through expression of the GM-CSF transgene.

2. 发明申请

US20150329873A1 FURIN-KNOCKDOWN AND GM-CSF-AUGMENTED (FANG) CANCER VACCINE 有权
标题翻译： FURIN-KNOCKDOWN和GM-CSF-AUGMENTED（FANG）CANCER VACCINE
公开(公告)号：US20150329873A1
公开(公告)日：2015-11-19
申请号：US14815721
申请日：2015-07-31
申请人： Gradalis, Inc.
发明人： John J. Nemunaitis , Neil Senzer , Phillip B. Maples , Donald Rao
IPC分类号： C12N15/85 , A61K38/21 , A61K35/13 , A61K39/00
CPC分类号： C12N15/85 , A61K31/7088 , A61K31/7105 , A61K35/13 , A61K38/217 , A61K39/0011
摘要： Compositions and methods for cancer treatment are disclosed herein. More specifically the present invention describes an autologous cancer vaccine genetically modified for Furin knockdown and GM-CSF expression. The vaccine described herein attenuates the immunosuppressive activity of TGF-β through the use of bi-functional shRNAs to knock down the expression of furin in cancer cells, and to augment tumor antigen expression, presentation, and processing through expression of the GM-CSF transgene.
摘要翻译：本文公开了用于癌症治疗的组合物和方法。更具体地，本发明描述了用于Furin敲低和GM-CSF表达遗传修饰的自体癌症疫苗。本文所述的疫苗减弱TGF-β的免疫抑制活性; 通过使用双功能shRNAs敲低癌细胞中弗林蛋白酶的表达，并通过GM-CSF转基因的表达增强肿瘤抗原的表达，表达和加工。

3. 发明申请

US20130259925A1 METHODS AND COMPOSITIONS TO TREAT CANCER USING BIFUNCTIONAL SRC 3 shRNA 审中-公开
标题翻译：使用双功能SRC 3 shRNA治疗癌症的方法和组成
公开(公告)号：US20130259925A1
公开(公告)日：2013-10-03
申请号：US13851464
申请日：2013-03-27
申请人： GRADALIS, INC. , BAYLOR COLLEGE OF MEDICINE
发明人： Donald Rao , Zhaohui Wang , John J. Nemunaitis , Neil Senzer , Bert W. O'Malley , David Lonard
IPC分类号： A61K31/713 , A61K45/06 , C12N15/113
CPC分类号： A61K31/713 , A61K45/06 , C12N15/113 , C12N15/1137 , C12N2330/51 , C12N2310/14 , C12N2310/531
摘要： The present invention includes compositions and methods of making and using an expression vector comprising a promoter and a nucleic acid insert operably linked to the promoter, wherein the insert encodes one or more short hairpin RNAs (shRNA) capable of inhibiting an expression of a SRC-3 gene via RNA interference, wherein the one or more shRNA comprise a bifunctional RNA molecule that activates a cleavage-dependent and a cleavage-independent RNA-induced silencing complex for reducing the expression level of the SRC-3.
摘要翻译：本发明包括制备和使用包含与启动子可操作地连接的启动子和核酸插入物的表达载体的组合物和方法，其中所述插入物编码一种或多种能够抑制SRC-表达的短发夹RNA（shRNA） 3基因，其中所述一个或多个shRNA包含激活切割依赖性的双功能RNA分子和用于降低SRC-3的表达水平的切割非依赖性RNA诱导的沉默复合物。

4. 发明申请

US20150368651A1 FURIN-KNOCKDOWN BI-FUNCTIONAL RNA 有权
标题翻译： FURIN-KNOCKDOWN双功能RNA
公开(公告)号：US20150368651A1
公开(公告)日：2015-12-24
申请号：US14844912
申请日：2015-09-03
申请人： GRADALIS, INC.
发明人： John J. Nemunaitis , Neil Senzer , Phillip B. Maples , Donald Rao
IPC分类号： C12N15/113 , A61K38/19 , A61K48/00 , A61K38/21 , C07K14/535
CPC分类号： C12N15/1137 , A61K38/193 , A61K38/217 , A61K48/005 , C07K14/535 , C12N15/85 , C12N2310/14 , C12N2310/141 , C12N2310/3519 , C12N2310/531 , C12N2320/31 , C12N2320/35 , C12N2330/51 , C12N2830/20 , C12N2840/107 , A61K2300/00
摘要： Compositions and methods to attenuate the immunosuppressive activity of TGF-β through the use of bi-functional shRNAs is described herein. The bi-functional shRNAs of the present invention knocks down the expression of furin in cancer cells to augment tumor antigen expression, presentation, and processing through expression of the GM-CSF transgene.
摘要翻译：降低TGF-β的免疫抑制活性的组合物和方法通过使用双功能shRNAs在本文中描述。本发明的双功能shRNAs通过表达GM-CSF转基因敲除癌细胞中弗林蛋白酶的表达，以增加肿瘤抗原的表达，呈递和加工。

5. 发明申请

US20140134236A1 INDIVIDUALIZED CANCER THERAPY 有权
标题翻译：个体化癌症治疗
公开(公告)号：US20140134236A1
公开(公告)日：2014-05-15
申请号：US14071490
申请日：2013-11-04
申请人： Gradalis, Inc.
发明人： David Shanahan , John J. Nemunaitis , Neil Senzer , Phillip B. Maples , Donald Rao
IPC分类号： C12Q1/68 , G01N33/574 , A61K31/713
CPC分类号： C12Q1/6886 , A61K31/7105 , A61K31/713 , A61K48/00 , C12N15/111 , C12N15/113 , C12Q2600/156 , C12Q2600/158 , C12Q2600/178 , G01N33/574 , G01N33/57484
摘要： In certain preferred embodiments, the invention provides methods for treating cancer, which comprise (a) obtaining a specimen of cancer tissue from a patient; (b) obtaining a specimen of normal tissue in the proximity of the cancer tissue from such patient; (c) extracting total protein and RNA from the cancer tissue and normal tissue; (d) obtaining a protein expression profile of the cancer tissue and normal tissue using 2D DIGE and mass spectrometry; (e) identifying proteins that are expressed in such cancer tissue at significantly different levels than in the normal tissue; (f) obtaining a gene expression profile of the cancer tissue and normal tissue using microarray technology and comparing the results thereof to the protein expression profile; (g) prioritizing over-expressed proteins by assessing the connectivity thereof to other cancer-related or stimulatory proteins; (h) designing an appropriate RNA interference expression cassette to, directly or indirectly, modulate the expression of genes encoding such prioritized proteins; (i) incorporating said cassette into an appropriate delivery vehicle; and (j) providing the patient with an effective amount of the delivery vehicle to, directly or indirectly, modify the expression (i.e., production) of such proteins.
摘要翻译：在某些优选实施方案中，本发明提供了治疗癌症的方法，其包括（a）从患者获得癌组织标本; （b）在患者的癌组织附近获得正常组织标本; （c）从癌组织和正常组织提取总蛋白和RNA; （d）使用2D DIGE和质谱法获得癌组织和正常组织的蛋白质表达谱; （e）以与正常组织中显着不同的水平鉴定在这样的癌组织中表达的蛋白质; （f）使用微阵列技术获得癌组织和正常组织的基因表达谱，并比较其结果与蛋白质表达谱; （g）通过评估与其他癌症相关或刺激性蛋白质的连通性来优先考虑过表达的蛋白质; （h）设计合适的RNA干扰表达盒直接或间接地调节编码这些优先蛋白质的基因的表达; （i）将所述盒装入合适的运送车辆; 和（j）向患者提供有效量的递送载体直接或间接地修饰这些蛋白质的表达（即生产）。

6. 发明申请

US20130266639A1 METHODS FOR TREATING TRIPLE NEGATIVE BREAST CANCER USING BIFUNCTIONAL SRC 3 shRNA 审中-公开
标题翻译：使用双功能SRC 3 shRNA治疗三重阴性乳腺癌的方法
公开(公告)号：US20130266639A1
公开(公告)日：2013-10-10
申请号：US13917499
申请日：2013-06-13
申请人： Baylor College of Medicine , Gradalis, Inc.
发明人： Donald Rao , Zhaohui Wang , John J. Nemunaitis , Neil Senzer , Bert W. O'Malley , David Lonard
IPC分类号： C12N15/113 , A61N5/10 , A61K45/06 , A61K31/138 , A61K9/127 , A61K31/713
CPC分类号： C12N15/1137 , A61K9/127 , A61K31/138 , A61K31/713 , A61K45/06 , A61N5/10 , C12N2310/51 , C12Y203/01048 , A61K2300/00 , C12N2310/14 , C12N2310/531
摘要： The present invention includes compositions and methods treating triple negative breast cancer comprising administering a therapeutically effective amount of a formulation that includes vector that expresses an SRC-1-specific bifunctional shRNA, an SRC-3-specific bifunctional shRNA, or both, to impair triple negative breast cancer cell growth.
摘要翻译：本发明包括治疗三阴性乳腺癌的组合物和方法，其包括施用治疗有效量的包含表达SRC-1特异性双功能shRNA，SRC-3特异性双功能shRNA或两者的载体的制剂，以损害三联体阴性乳腺癌细胞生长。

7. 发明申请

US20150368640A9 Novel shRNA Molecules and Methods of Use Thereof 有权
标题翻译：新型shRNA分子及其使用方法
公开(公告)号：US20150368640A9
公开(公告)日：2015-12-24
申请号：US14318163
申请日：2014-06-27
申请人： Gradalis, Inc.
发明人： Donald Rao
IPC分类号： C12N15/113
CPC分类号： C12N15/113 , A61K48/00 , C07H21/02 , C12N15/111 , C12N15/85 , C12N2310/14 , C12N2310/531 , C12N2320/30 , C12N2320/31
摘要： The present invention relates to certain novel shRNA molecules and methods of use thereof. According to certain embodiments of the present invention, methods for reducing the expression level of a target gene are provided. Such methods generally comprise providing a cell with one or more precursor nucleic acid sequences that encode two or more RNA molecules. A first RNA molecule comprises a double stranded sequence, which includes a guide strand sequence that is complementary to a portion of an mRNA transcript encoded by the target gene. In addition, a second RNA molecule comprises a second double stranded sequence, which includes a second guide strand sequence that is partially complementary to a portion of the mRNA transcript encoded by the target gene. Preferably, the second guide strand sequence comprises one or more bases that are mismatched with a nucleic acid sequence of the mRNA transcript encoded by the target gene.
摘要翻译：本发明涉及某些新型shRNA分子及其使用方法。根据本发明的某些实施方案，提供降低靶基因表达水平的方法。这样的方法通常包括向细胞提供编码两个或更多个RNA分子的一个或多个前体核酸序列。第一RNA分子包含双链序列，其包括与靶基因编码的mRNA转录物的一部分互补的引导链序列。此外，第二RNA分子包含第二双链序列，其包括与由靶基因编码的mRNA转录物的一部分部分互补的第二引导链序列。优选地，第二引导链序列包含与靶基因编码的mRNA转录物的核酸序列错配的一个或多个碱基。

8. 发明申请

US20140242639A1 CONSTRUCTION OF BIFUNCTIONAL SHORT HAIRPIN RNA 有权
标题翻译：双功能短发HAIRPIN RNA的构建
公开(公告)号：US20140242639A1
公开(公告)日：2014-08-28
申请号：US14271039
申请日：2014-05-06
申请人： Gradalis, Inc.
发明人： Donald Rao
IPC分类号： C12N15/113
CPC分类号： C12N15/113 , A61K9/0019 , A61K48/00 , C12N15/111 , C12N15/85 , C12N2310/14 , C12N2310/51 , C12N2310/53 , C12N2330/51 , C12P19/34 , C12N2310/531
摘要： A method for designing a bi-shRNA expression cassette encoding a bi-shRNA comprising: selecting one or more target site sequences; providing a backbone sequence comprising a first and a second stem-loop structure, inserting a first passenger strand and a second passenger strand and providing for synthesis of the bi-shRNA expression cassette.
摘要翻译：一种编码双shRNA的双shRNA表达盒的设计方法，包括：选择一个或多个靶位点序列; 提供包括第一和第二茎环结构的骨架序列，插入第一乘客链和第二乘客链，并提供合成双shRNA表达盒。

9. 发明申请

US20130259926A1 BI-FUNCTIONAL shRNA TARGETING MESOTHELIN AND USES THEREOF 审中-公开
标题翻译：双功能shRNA靶向MESOTHELIN及其用途
公开(公告)号：US20130259926A1
公开(公告)日：2013-10-03
申请号：US13852446
申请日：2013-03-28
申请人： GRADALIS, INC. , BAYLOR COLLEGE OF MEDICINE
发明人： Donald Rao , Zhaohui Wang , John J. Nemunaitis , Neil Senzer , Qizhi Yao , Changyi (Johnny) Chen , F. Charles Brunicardi
IPC分类号： C12N15/113
CPC分类号： C12N15/1138 , C12N2310/51 , C12N2310/14 , C12N2310/531
摘要： The present invention includes bifunctional shRNAs capable of reducing an expression of a Mesothelin gene; wherein at least one target site sequence of the bifunctional RNA molecule is located within the Mesothelin, wherein the bifunctional RNA molecule is capable of activating a cleavage-dependent and a cleavage-independent RNA-induced silencing complex for reducing the expression level of Mesothelin.
摘要翻译：本发明包括能够降低间皮素基因表达的双功能shRNA; 其中所述双功能RNA分子的至少一个靶位点序列位于间皮素内，其中所述双功能RNA分子能够激活切割依赖性和切割独立的RNA诱导的沉默复合物，以降低间皮素的表达水平。

10. 发明授权

US09914977B2 Individualized cancer therapy 有权
公开(公告)号：US09914977B2
公开(公告)日：2018-03-13
申请号：US15173395
申请日：2016-06-03
申请人： GRADALIS, INC.
发明人： David M. Shanahan , John J. Nemunaitis , Neil Senzer , Phillip B. Maples , Donald Rao
IPC分类号： A61K31/70 , C07H21/02 , C07H21/04 , C12Q1/68 , C12N15/113 , C12N15/11 , A61K31/7105 , A61K48/00 , G01N33/574 , A61K31/713 , C12N5/00
CPC分类号： C12Q1/6886 , A61K31/7105 , A61K31/713 , A61K48/00 , C12N15/111 , C12N15/113 , C12Q2600/156 , C12Q2600/158 , C12Q2600/178 , G01N33/574 , G01N33/57484
摘要： In certain preferred embodiments, the invention provides methods for treating cancer, which comprise (a) obtaining a specimen of cancer tissue from a patient; (b) obtaining a specimen of normal tissue in the proximity of the cancer tissue from such patient; (c) extracting total protein and RNA from the cancer tissue and normal tissue; (d) obtaining a protein expression profile of the cancer tissue and normal tissue using 2D DIGE and mass spectrometry; (e) identifying proteins that are expressed in such cancer tissue at significantly different levels than in the normal tissue; (f) obtaining a gene expression profile of the cancer tissue and normal tissue using microarray technology and comparing the results thereof to the protein expression profile; (g) prioritizing over-expressed proteins by assessing the connectivity thereof to other cancer-related or stimulatory proteins; (h) designing an appropriate RNA interference expression cassette to, directly or indirectly, modulate the expression of genes encoding such prioritized proteins; (i) incorporating said cassette into an appropriate delivery vehicle; and (j) providing the patient with an effective amount of the delivery vehicle to, directly or indirectly, modify the expression (i.e., production) of such proteins.

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