JP5171887B2 - 癌関連タンパク質を標的とするRNAiプローブ - Google Patents
癌関連タンパク質を標的とするRNAiプローブ Download PDFInfo
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- JP5171887B2 JP5171887B2 JP2010128382A JP2010128382A JP5171887B2 JP 5171887 B2 JP5171887 B2 JP 5171887B2 JP 2010128382 A JP2010128382 A JP 2010128382A JP 2010128382 A JP2010128382 A JP 2010128382A JP 5171887 B2 JP5171887 B2 JP 5171887B2
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/712—Nucleic acids or oligonucleotides having modified sugars, i.e. other than ribose or 2'-deoxyribose
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/04—Antineoplastic agents specific for metastasis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/31—Chemical structure of the backbone
- C12N2310/315—Phosphorothioates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/32—Chemical structure of the sugar
- C12N2310/321—2'-O-R Modification
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/34—Spatial arrangement of the modifications
- C12N2310/341—Gapmers, i.e. of the type ===---===
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/34—Spatial arrangement of the modifications
- C12N2310/346—Spatial arrangement of the modifications having a combination of backbone and sugar modifications
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Biotechnology (AREA)
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- Wood Science & Technology (AREA)
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- General Engineering & Computer Science (AREA)
- Epidemiology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
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- Oncology (AREA)
- Hospice & Palliative Care (AREA)
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
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Description
1)細胞の調製
抗生物質(ペニシリン/ストレプトマイシン)を含まない5%のFBSを含む適当な培地中のLNCaP細胞0.5×106個を(PC3細胞では、ウェル当たり0.3×106個の密度で)6穴プレートの各ウェルに播く。
40〜50%コンフルエントになるまで、細胞を37ECの加湿下5%CO2インキュベータ内でインキュベートする。
2)siRNAの調製
以下のsiRNA希釈液を微量遠心チューブに調製する。各ウェル当たり、0.01〜100nM。
3)以下のトランスフェクション試薬希釈液を微量遠心チューブに調製する。
6穴プレートの各ウェルについて、OligoFECTAMINE(商標)試薬4mlをOPTI−MEM(商標)11mlで希釈し、室温で10分間インキュベートする。
4)希釈したOligoFECTAMINE(商標)を、希釈したsiRNA二本鎖希釈液と合わせて、反転により穏やかに混合する。
5)室温で20分インキュベートする。
6)ウェルから培地を取り除き、Opti−MEM(商標)800mlと交換する。
7)細胞にトランスフェクション複合体200mlを上層する。
8)37℃のCO2インキュベータ内で4時間インキュベートする。
9)15%FBSを含む培地500mlを添加する。
10)24時間後にリアルタイムPCRにより遺伝子発現をチェックするか、又は
11)1、6、12、24、48、72及び96時間後にウエスタンブロットによりタンパク質発現をチェックする。
Claims (6)
- 各々の鎖が18〜23塩基長を有し、標的遺伝子の転写産物であるmRNAの分解を仲介するか翻訳を阻害するのに有効な配列を有する二重鎖のRNA分子であって、前記標的遺伝子が、インスリン様成長因子結合タンパク質−2(IGFBP−2)及びインスリン様成長因子結合タンパク質−5(IGFBP−5)の両方であり、二重鎖RNA分子の鎖が、
配列番号39及び配列番号40;
配列番号41及び配列番号42;
配列番号43及び配列番号44;
からなる群から選ばれる相補配列の対を含む、上記RNA分子。 - 前記RNA分子が
配列番号39及び配列番号40;
配列番号41及び配列番号42;
配列番号43及び配列番号44;
の中から選ばれる相補配列の対から成る、請求項1に記載のRNA分子。 - 請求項1又は2に記載のRNA分子及び薬学的に許容される担体を含む医薬品組成物。
- 前記の薬学的に許容される担体が無菌注射液である請求項3に記載の医薬品組成物。
- インスリン様成長因子結合タンパク質−2(IGFBP−2)、インスリン様成長因子結合タンパク質−5(IGFBP−5)、又はIGFBP−2及びIGFBP−5の両方を発現する癌の治療のための医薬組成物であって、有効成分として請求項1又は2に記載のRNA分子を含有する上記医薬組成物。
- 前記癌が、前立腺癌又は乳癌である、請求項5に記載の医薬組成物。
Applications Claiming Priority (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US40519302P | 2002-08-21 | 2002-08-21 | |
US60/405,193 | 2002-08-21 | ||
US40815202P | 2002-09-03 | 2002-09-03 | |
US60/408,152 | 2002-09-03 | ||
US47238703P | 2003-05-20 | 2003-05-20 | |
US60/472,387 | 2003-05-20 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2005501198A Division JP4717633B2 (ja) | 2002-08-21 | 2003-08-21 | 癌関連タンパク質を標的とするRNAiプローブ |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2010193908A JP2010193908A (ja) | 2010-09-09 |
JP5171887B2 true JP5171887B2 (ja) | 2013-03-27 |
Family
ID=31950539
Family Applications (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2005501198A Expired - Fee Related JP4717633B2 (ja) | 2002-08-21 | 2003-08-21 | 癌関連タンパク質を標的とするRNAiプローブ |
JP2010128382A Expired - Fee Related JP5171887B2 (ja) | 2002-08-21 | 2010-06-04 | 癌関連タンパク質を標的とするRNAiプローブ |
JP2010250838A Expired - Fee Related JP5662766B2 (ja) | 2002-08-21 | 2010-11-09 | 癌関連タンパク質を標的とするRNAiプローブ |
JP2013067754A Pending JP2013150624A (ja) | 2002-08-21 | 2013-03-28 | 癌関連タンパク質を標的とするRNAiプローブ |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2005501198A Expired - Fee Related JP4717633B2 (ja) | 2002-08-21 | 2003-08-21 | 癌関連タンパク質を標的とするRNAiプローブ |
Family Applications After (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2010250838A Expired - Fee Related JP5662766B2 (ja) | 2002-08-21 | 2010-11-09 | 癌関連タンパク質を標的とするRNAiプローブ |
JP2013067754A Pending JP2013150624A (ja) | 2002-08-21 | 2013-03-28 | 癌関連タンパク質を標的とするRNAiプローブ |
Country Status (10)
Country | Link |
---|---|
US (5) | US8252918B2 (ja) |
EP (2) | EP1532249A2 (ja) |
JP (4) | JP4717633B2 (ja) |
KR (3) | KR101212512B1 (ja) |
AU (1) | AU2003258426B2 (ja) |
CA (2) | CA2882443C (ja) |
IL (3) | IL166658A (ja) |
NO (2) | NO334573B1 (ja) |
NZ (1) | NZ552872A (ja) |
WO (1) | WO2004018676A2 (ja) |
Families Citing this family (26)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
HU227190B1 (en) | 1999-02-26 | 2010-10-28 | Univ British Columbia | Trpm-2 antisense therapy |
US7569551B2 (en) | 2000-02-25 | 2009-08-04 | The University Of British Columbia | Chemo- and radiation-sensitization of cancer by antisense TRPM-2 oligodeoxynucleotides |
ATE402999T1 (de) | 2002-01-17 | 2008-08-15 | Univ British Columbia | Bispezifische antisense oligonukleotide die igfbp-2 und igfbp-5 inhibieren und deren verwendung |
CA2882443C (en) * | 2002-08-21 | 2016-12-13 | The University Of British Columbia | Rnai probes targeting cancer-related proteins |
ATE478142T1 (de) * | 2002-08-21 | 2010-09-15 | Univ British Columbia | Behandlung von melanomen durch reduktion der clusterin menge |
US7635673B2 (en) | 2003-03-25 | 2009-12-22 | The Board Of Trustees Of The University Of Illinois | Methods of inhibiting tumor cell proliferation |
US20040220131A1 (en) * | 2003-04-18 | 2004-11-04 | The University Of British Columbia | Method for treatment of cancerous angiogenic disorders |
US20050019918A1 (en) * | 2003-06-03 | 2005-01-27 | Hidetoshi Sumimoto | Treatment of cancer by inhibiting BRAF expression |
JP2005013221A (ja) * | 2003-06-03 | 2005-01-20 | Keio Gijuku | Braf発現抑制を利用した癌の治療 |
MXPA06010667A (es) * | 2004-03-19 | 2007-07-04 | Penn State Res Found | Metodos combinatorios y composiciones para el tratamiento de melanoma. |
US8710020B2 (en) | 2004-04-02 | 2014-04-29 | The University Of British Columbia | Clusterin antisense therapy for treatment of cancer |
EP2298896A1 (en) * | 2004-06-22 | 2011-03-23 | The Board of Trustees of the University of Illinois | Methods of inhibiting tumor cell proliferation with FOXM1 siRNA |
TWI661199B (zh) * | 2004-07-23 | 2019-06-01 | 太平洋愛吉生技股份有限公司 | 檢測膀胱癌之尿液標記物 |
WO2006035432A2 (en) * | 2004-09-27 | 2006-04-06 | Tel Hashomer Medical Research Infrastructure And Services Ltd. | Gene silencing for use in dermatology |
EP1814595B1 (en) * | 2004-11-23 | 2014-01-08 | The University Of British Columbia | Treatment of cancer with a combination of an agent that perturbs the egf signaling pathway and an oligonucleotide that reduces clusterin levels |
EP1937815B1 (en) * | 2005-09-13 | 2015-05-13 | National Research Council of Canada | Methods and compositions for modulating tumor cell activity |
US8029980B2 (en) | 2006-09-29 | 2011-10-04 | The Board Of Trustees Of The University Of Illinois | Identification and use of agents that modulate oncogenic transcription agent activity |
EP2117557A4 (en) * | 2007-01-16 | 2011-01-19 | Burnham Inst Medical Research | COMPOSITIONS AND METHODS FOR THE TREATMENT OF COLORECTAL CANCER |
WO2008094516A2 (en) * | 2007-01-29 | 2008-08-07 | City Of Hope | Multi-targeting short interfering rnas |
EP2121923A1 (en) * | 2007-03-02 | 2009-11-25 | MDRNA, Inc. | Nucleic acid compounds for inhibiting erbb family gene expression and uses thereof |
US8217161B2 (en) * | 2008-04-22 | 2012-07-10 | Clemson University Research Foundation | Methods of inhibiting multiple cytochrome P450 genes with siRNA |
DK2504363T3 (da) | 2009-11-24 | 2019-07-29 | Alethia Biotherapeutics Inc | Anti-clusterin-antistoffer og antigenbindende fragmenter og deres anvendelse til reducering af tumorvolumen |
EP3301176B1 (en) | 2011-02-11 | 2019-09-25 | The Rockefeller University | Method for identifying nucleic acids regulating metastasation |
JP2015512877A (ja) | 2012-02-22 | 2015-04-30 | アレシア・バイオセラピューティクス・インコーポレーテッド | 癌を治療するためのクラステリン阻害剤とegfr阻害剤の併用 |
CA2844640A1 (en) | 2013-12-06 | 2015-06-06 | The University Of British Columbia | Method for treatment of castration-resistant prostate cancer |
CN106282185B (zh) * | 2016-08-18 | 2020-06-26 | 广州市锐博生物科技有限公司 | 一种用于抑制簇集蛋白基因表达的成套siRNA及其应用 |
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JPS5736580A (en) * | 1980-08-13 | 1982-02-27 | Hitachi Ltd | Protecting method for converter |
AU663702B2 (en) * | 1991-03-06 | 1995-10-19 | Board Of Regents, The University Of Texas System | Methods and compositions for the selective inhibition of gene expression |
AUPM672594A0 (en) * | 1994-07-08 | 1994-08-04 | Royal Children's Hospital Research Foundation | A method for the prophylaxis and/or treatment of proliferative and/or inflammatory skin disorders |
US20020086386A1 (en) * | 1997-03-04 | 2002-07-04 | Kamb Carl Alexander | B-catenin assays, and compositions therefrom |
US6383808B1 (en) | 2000-09-11 | 2002-05-07 | Isis Pharmaceuticals, Inc. | Antisense inhibition of clusterin expression |
HU227190B1 (en) * | 1999-02-26 | 2010-10-28 | Univ British Columbia | Trpm-2 antisense therapy |
WO2000069454A1 (en) * | 1999-05-17 | 2000-11-23 | Board Of Regents, The University Of Texas System | Suppression of endogenous igfbp-2 to inhibit cancer |
AU772480B2 (en) | 1999-07-19 | 2004-04-29 | University Of British Columbia, The | Antisense therapy for hormone-regulated tumors |
US6140126A (en) * | 1999-10-26 | 2000-10-31 | Isis Pharmaceuticals Inc. | Antisense modulation of Y-box binding protein 1 expression |
GB9927444D0 (en) | 1999-11-19 | 2000-01-19 | Cancer Res Campaign Tech | Inhibiting gene expression |
US7569551B2 (en) | 2000-02-25 | 2009-08-04 | The University Of British Columbia | Chemo- and radiation-sensitization of cancer by antisense TRPM-2 oligodeoxynucleotides |
WO2001068836A2 (en) * | 2000-03-16 | 2001-09-20 | Genetica, Inc. | Methods and compositions for rna interference |
JP5500750B2 (ja) | 2000-03-30 | 2014-05-21 | ホワイトヘッド インスチチュート フォアー バイオメディカル リサーチ | Rna干渉のrna配列特異的メディエータ |
HU229207B1 (en) | 2000-09-14 | 2013-09-30 | Univ British Columbia | Antisense insulin-like growth factor binding protein (igfbp)-2-oligodeoxynucleotides for prostate and other endocrine regulated tumor therapy |
US20040259247A1 (en) * | 2000-12-01 | 2004-12-23 | Thomas Tuschl | Rna interference mediating small rna molecules |
EP1354038A2 (en) | 2000-12-28 | 2003-10-22 | J & J Research Pty Ltd | Double-stranded rna-mediated gene suppression |
EP1386004A4 (en) * | 2001-04-05 | 2005-02-16 | Ribozyme Pharm Inc | MODULATION OF GENE EXPRESSION ASSOCIATED WITH INFLAMMATORY PROLIFERATION AND GROWTH OF NEURITIES BY METHODS INVOLVING NUCLEIC ACID |
ATE402999T1 (de) * | 2002-01-17 | 2008-08-15 | Univ British Columbia | Bispezifische antisense oligonukleotide die igfbp-2 und igfbp-5 inhibieren und deren verwendung |
CA2882443C (en) * | 2002-08-21 | 2016-12-13 | The University Of British Columbia | Rnai probes targeting cancer-related proteins |
ATE478142T1 (de) | 2002-08-21 | 2010-09-15 | Univ British Columbia | Behandlung von melanomen durch reduktion der clusterin menge |
US7250496B2 (en) * | 2002-11-14 | 2007-07-31 | Rosetta Genomics Ltd. | Bioinformatically detectable group of novel regulatory genes and uses thereof |
EP2305812A3 (en) * | 2002-11-14 | 2012-06-06 | Dharmacon, Inc. | Fuctional and hyperfunctional sirna |
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2003
- 2003-08-21 CA CA2882443A patent/CA2882443C/en not_active Expired - Fee Related
- 2003-08-21 AU AU2003258426A patent/AU2003258426B2/en not_active Ceased
- 2003-08-21 KR KR1020057002963A patent/KR101212512B1/ko not_active IP Right Cessation
- 2003-08-21 KR KR1020107028249A patent/KR101117673B1/ko not_active IP Right Cessation
- 2003-08-21 US US10/646,436 patent/US8252918B2/en not_active Expired - Fee Related
- 2003-08-21 WO PCT/CA2003/001277 patent/WO2004018676A2/en active Application Filing
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- 2003-08-21 EP EP03792075A patent/EP1532249A2/en not_active Ceased
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- 2003-08-21 KR KR1020117025408A patent/KR101238701B1/ko not_active IP Right Cessation
- 2003-08-21 EP EP10010458.7A patent/EP2263679B1/en not_active Expired - Lifetime
- 2003-08-21 CA CA2494766A patent/CA2494766C/en not_active Expired - Fee Related
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2005
- 2005-02-02 IL IL166658A patent/IL166658A/en not_active IP Right Cessation
- 2005-03-17 NO NO20051423A patent/NO334573B1/no not_active IP Right Cessation
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2006
- 2006-02-23 US US11/276,300 patent/US7964717B2/en not_active Expired - Fee Related
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2008
- 2008-05-06 US US12/116,083 patent/US7820635B2/en not_active Expired - Fee Related
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2009
- 2009-02-22 IL IL197159A patent/IL197159A/en not_active IP Right Cessation
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2010
- 2010-06-04 JP JP2010128382A patent/JP5171887B2/ja not_active Expired - Fee Related
- 2010-07-28 US US12/845,521 patent/US8759308B2/en not_active Expired - Fee Related
- 2010-11-09 JP JP2010250838A patent/JP5662766B2/ja not_active Expired - Fee Related
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2012
- 2012-02-24 US US13/404,741 patent/US9487777B2/en not_active Expired - Fee Related
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