JP2001517939A - ペプチドによって増強されるトランスフェクション - Google Patents
ペプチドによって増強されるトランスフェクションInfo
- Publication number
- JP2001517939A JP2001517939A JP53989998A JP53989998A JP2001517939A JP 2001517939 A JP2001517939 A JP 2001517939A JP 53989998 A JP53989998 A JP 53989998A JP 53989998 A JP53989998 A JP 53989998A JP 2001517939 A JP2001517939 A JP 2001517939A
- Authority
- JP
- Japan
- Prior art keywords
- peptide
- transfection
- composition
- protein
- nucleic acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
- A61K47/543—Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
- A61K47/645—Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6949—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
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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
- 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
- A61K48/0008—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
- A61K48/0025—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition wherein the non-active part clearly interacts with the delivered nucleic acid
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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
- A61K48/0008—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
- A61K48/0025—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition wherein the non-active part clearly interacts with the delivered nucleic acid
- A61K48/0041—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition wherein the non-active part clearly interacts with the delivered nucleic acid the non-active part being polymeric
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
- A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers
- A61K9/1272—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
- C07K1/1072—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides by covalent attachment of residues or functional groups
- C07K1/1077—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides by covalent attachment of residues or functional groups by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2740/00—Reverse transcribing RNA viruses
- C12N2740/00011—Details
- C12N2740/10011—Retroviridae
- C12N2740/16011—Human Immunodeficiency Virus, HIV
- C12N2740/16311—Human Immunodeficiency Virus, HIV concerning HIV regulatory proteins
- C12N2740/16322—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Engineering & Computer Science (AREA)
- Nanotechnology (AREA)
- Wood Science & Technology (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- General Chemical & Material Sciences (AREA)
- Medical Informatics (AREA)
- Plant Pathology (AREA)
- Gastroenterology & Hepatology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Virology (AREA)
- Dispersion Chemistry (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Peptides Or Proteins (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
Claims (1)
- 【特許請求の範囲】 1.1つ以上の核酸分子、1つ以上のペプチドまたはタンパク質、および1つ以 上のトランスフェクション薬剤を含む、細胞をトランスフェクトするための組成 物。 2.前記組成物が、2つ以上のペプチドおよび/またはタンパク質を含む、請求 項1に記載の組成物。 3.前記組成物が、2つ以上のトランスフェクション薬剤を含む、請求項1に記 載の組成物。 4.前記組成物が、ペプチド−核酸複合体またはタンパク質−核酸複合体を含む 、請求項1に記載の組成物。 5.前記ペプチド−核酸複合体またはタンパク質−核酸複合体が、2つ以上のペ プチドもしくはタンパク質または両方を含む、請求項4に記載の組成物。 6.前記トランスフェクション薬剤が、1つ以上のカチオン性脂質を含む、請求 項1に記載の組成物。 7.前記トランスフェクション薬剤が、さらに1つ以上の中性脂質を含む、請求 項6に記載の組成物。 8.前記トランスフェクション薬剤が、1つ以上のデンドリマーを含む、請求項 1に記載の組成物。 9.前記トランスフェクション薬剤が、さらに1つ以上の脂質を含む、請求項8 に記載の組成物。 10.前記カチオン性脂質が、1つ以上の1価カチオン性脂質を含む、請求項6 に記載の組成物。 11.前記1価カチオン性脂質が、DOTMA、DOTAP、DMRIE、およびDDABからなる 群から選択される、請求項10に記載の組成物。 12.前記カチオン性脂質が、1つ以上の多価カチオン性脂質を含む、請求項6 に記載の組成物。 13.前記多価カチオン性脂質が、DOSPA、DOSPER、DOGS、TMTPS、TMTOS、TMTLS 、TMTMS、およびTMDOSからなる群から選択される、請求項12に記載の組成物。 14.前記中性脂質が、DOPE、DPhPE、およびコレステロールからなる群から選 択される、請求項7に記載の組成物。 15.前記デンドリマーが、デンススターデンドリマー、PAMAMデンドリマー、N H3コアデンドリマー、エチレンジアミンコアデンドリマー、ジェネレーション5 以上のデンドリマー、置換基を有するデンドリマー、1つ以上のアミノ酸を含む デンドリマー、グラフト化デンドリマー、および活性化されたデンドリマーから なる群から選択される、請求項8に記載の組成物。 16.1つ以上の前記トランスフェクション薬剤が、1つ以上の前記ペプチドお よび/またはタンパク質に共有結合している、請求項1に記載の組成物。 17.1つ以上の前記カチオン性脂質が、1つ以上の前記ペプチドおよび/また はタンパク質に共有結合している、請求項6に記載の組成物。 18.1つ以上の前記中性脂質が、1つ以上の前記ペプチドおよび/またはタン パク質に共有結合している、請求項7に記載の組成物。 19.1つ以上の前記デンドロマーが、1つ以上の前記ペプチドおよび/または タンパク質に共有結合している、請求項8に記載の組成物。 20.前記ペプチドおよび/またはタンパク質が、動物、細菌、ウイルスのペプ チドおよび/またはタンパク質由来である、請求項1に記載の組成物。 21.前記ペプチドおよび/またはタンパク質が、1つ以上の核酸結合基と結合 している、請求項1に記載の組成物。 22.前記核酸結合基が、少なくとも1つのポリアミンを含む、請求項21に記 載の組成物。 23.前記核酸結合基が、少なくとも1つのスペルミンを含む、請求項22に記 載の組成物。 24.少なくとも1つの前記ペプチドおよび/またはタンパク質が、核局在化タ ンパク質またはペプチドである、請求項1に記載の組成物。 25.少なくとも1つの前記ペプチドおよび/またはタンパク質が、融合誘導ペ プチドまたはタンパク質である、請求項1に記載の組成物。 26.少なくとも1つの前記ペプチドおよび/またはタンパク質が、レセプター −リガンドペプトドまたはタンパク質である、請求項1に記載の組成物。 27.少なくとも1つの前記ペプチドおよび/またはタンパク質が、輸送ペプチ ドまたはタンパク質である、請求項1に記載の組成物。 28.少なくとも1つの前記ペプチドおよび/またはタンパク質が、ウイルス性 ペプチドまたはタンパク質である、請求項20に記載の組成物。 29.前記ウイルスが、インフルエンザウイルス、水疱性口内炎ウイルス、アデ ノウイルス、アルファウイルス、セムリキ森林ウイルス、肝炎ウイルス、ヘルペ スウイルス、HIVウイルス、およびシミアンウイルスからなる群から選択される 、請求項28に記載の組成物。 30.さらにDEAE-デキストラン、クロロキン、またはその組み合わせを含む、 請求項1に記載の組成物。 31.少なくとも1つの前記ペプチドおよび/またはタンパク質が、インスリン 、トランスフェリン、上皮増殖因子、線維芽細胞増殖因子、ラクトフェリン、フ ィブロネクチン、アデノウイルスペントンベース、Knob、およびヘキソンタンパ ク質、水疱性口内炎ウイルス糖タンパク質、セムリキ森林ウイルスコアタンパク 質、インフルエンザ赤血球凝集素、B型肝炎コアタンパク質、HIV Tatタンパク質 、単純ヘルペスウイルスVP22タンパク質、ヒストンタンパク質、HMGタンパク質 、およびインベーシンタンパク質、インターナリンタンパク質、内毒素、ジフテ リア毒素、赤痢菌毒素、メリチン、マガイニン、グラミシジン、セクロフィン、 ディフェンシン類、プロテグリン類、タキプレシン類、チオニン類、インドリシ ジン、バクテネシン、ドロソマイシン、アピダエシン類、カテリシジン、殺菌性 透過性増加タンパク質、ナイシン、ブホリン、およびそのフラグメントからなる 群から選択される、請求項1に記載の組成物。 32.前記組成物が、初代細胞培養物、継代細胞培養物、または細胞株をトラン スフェクトし得る、請求項1に記載の組成物。 33.前記細胞株が、ヒト細胞株である、請求項32に記載の組成物。 34.前記細胞株が、動物細胞株である、請求項32に記載の組成物。 35.前記細胞株が、線維芽細胞である、請求項32に記載の組成物。 36.少なくとも1つの前記ペプチドおよび/またはタンパク質が、同じもしく は異なるペプチドまたはタンパク質の多量体を含む、請求項1に記載の組成物。 37.前記ペプチドおよび/またはタンパク質が、1つ以上のアミノ酸誘導体ま たはアナログを含む、請求項1に記載の組成物。 38.少なくとも1つの前記ペプチドおよび/またはタンパク質が、融合誘導、 核局在化、輸送、レセプター-リガンドおよび細胞接着からなる群から選択され る、2つ以上の機能を含む、請求項1に記載の組成物。 39.標的細胞または組織のトランスフェクションに有効な量の請求項1に記載 の組成物および薬学的キャリアを含む、薬学的組成物。 40.選択された治療用核酸を有する、標的細胞または組織のトランスフェクシ ョンに有効な量の請求項1に記載の組成物を含む、治療用組成物。 41.選択された診断用核酸を有する、標的細胞または組織のトランスフェクシ ョンに有効な量の請求項1に記載の組成物を含む、診断用組成物。 42.ペプチドまたはタンパク質に共有結合したトランスフェクション薬剤の成 分を含む、細胞をトランスフェクトするための組成物。 43.前記トランスフェクション薬剤の成分が脂質である、請求項42に記載の 組成物。 44.前記トランスフェクション薬剤の成分がカチオン性脂質である、請求項4 2に記載の組成物。 45.前記トランスフェクション薬剤の成分が中性脂質である、請求項42に記 載の組成物。 46.前記トランスフェクション薬剤の成分がデンドリマーである、請求項42 に記載の組成物。 47.レセプター-リガンドタンパク質をさらに含む、請求項42に記載の組成 物。 48.1つ以上の核酸分子、1つ以上のペプチドまたはタンパク質、および1つ 以上のトランスフェクション薬剤の組み合わせによって得られる、細胞をトラン スフェクトするための組成物。 49.最初にペプチド-核酸複合体またはタンパク質-核酸複合体を形成し、続い て該ペプチド-核酸複合体またはタンパク質-核酸複合体と凝集し得るトランスフ ェクション薬剤を添加することによって得られる、請求項48に記載の細胞をト ランスフェクトするための組成物。 50.前記ペプチド核酸複合体またはタンパク質核酸複合体が形成された後に、 該複合体がカチオン性脂質および中性脂質の混合物に添加される、請求項49に 記載の組成物。 51.核酸を用いて細胞をトランスフェクトする方法であって、請求項1に記載 のトランスフェクション組成物を該細胞と接触させる工程を包含する、方法。 52.核酸を用いて細胞をトランスフェクトする方法であって、請求項17に記 載のトランスフェクション組成物を該細胞と接触させる工程を包含する、方法。 53.核酸を用いて細胞をトランスフェクトする方法であって、請求項31に記 載のトランスフェクション組成物を該細胞と接触させる工程を包含する、方法。 54.核酸を用いて細胞をトランスフェクトする方法であって、請求項48に記 載のトランスフェクション組成物を該細胞と接触させる工程を包含する、方法。 55.核酸を用いて細胞をトランスフェクトする方法であって、以下の工程: (a)1つ以上のペプチドまたはタンパク質を核酸と混合し、ペプチド-核酸 複合体またはタンパク質-核酸複合体を形成する工程; (b)工程(a)由来の複合体にトランスフェクション薬剤を添加し、該トラ ンスフェクション薬剤および該複合体の凝集体を得る工程;ならびに (c)該細胞を工程(b)由来の凝集体と接触させる工程、 を包含する、方法。 56.前記ペプチドまたはタンパク質が、細胞内局在化シグナル配列、核局在化 シグナル配列、融合誘導配列、輸送またはトラフィッキング配列、レセプター- リガンド配列、または細胞接着配列を含む、請求項55に記載の方法。 57.前記ペプチドまたはタンパク質が、核酸結合基への共有結合によって改変 される、請求項56に記載の方法。 58.前記核酸結合基が、スペルミンである、請求項57に記載の方法。 59.前記ペプチドが、Sp-NLS、Sp-NLSNLS、SP-NLSRGD、Opf-GG-1、Opf-GG-2、 Opf-GG-2-CYS、またはSP-Tatである、請求項58に記載の方法。 60.前記トランスフェクション薬剤がデンドリマーを含む、請求項55に記載 の方法。 61.前記トランスフェクション薬剤が活性化デンドリマーを含む、請求項60 に記載の方法。 62.前記デンドリマーが、GX(NH3)またはGX(EDA)デンドリマーからなる群から 選択され、ここでXは約5から約10の間の整数である、請求項61に記載の方 法。 63.前記デンドリマーがアルギニンまたはリジンに結合している、請求項60 に記載の方法。 64.トランスフェクション薬剤、およびトランスフェクション薬剤のトランス フェクションを増強し得るペプチドもしくはタンパク質、または改変ペプチドも しくはタンパク質を含む、トランスフェクション試薬キット。 65.カチオン性脂質トランスフェクション薬剤を含む、請求項64に記載のキ ット。 66.前記カチオン性脂質トランスフェクション薬剤が、「LIPOFECTAMINE」、 「LIPOFECTIN」、「LIPOFECTACE」、「CELLFECTIN」、「MULTIFECTOR」または「 TRANSFECTIN」からなる群から選択される、請求項65に記載のキット。 67.前記ペプチドがSp-NLSNLSである、請求項66に記載のキット。 68.前記ペプチドがSp-Tatである、請求項66に記載のキット。 69.デンドリマートランスフェクション薬剤を含む、請求項64に記載のキッ ト。 70.前記デンドリマーがデンススターデンドリマーまたは活性化デンドリマー である、請求項69に記載のキット。 71.診断用キットであって、診断用核酸をさらに含む、請求項64に記載のキ ット。 72.核酸結合基への共有結合によって改変されるNLS配列を含む、ペプチド。 73.NLS配列のダイマーまたは多量体を含む、請求項72に記載の改変ペプチ ド。 74.前記核酸結合基がスペルミンである、請求項73に記載の改変ペプチド。 75.核酸結合基への共有結合によって改変された、Tat配列を含む、ペプチド 。 76.Tat配列のダイマーまたは多量体を含む、請求項75に記載の改変ペプチ ド。 77.前記核酸結合基がスペルミンである、請求項76に記載の改変ペプチド。
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Also Published As
Publication number | Publication date |
---|---|
EP2206787A3 (en) | 2010-12-01 |
WO1998040502A1 (en) | 1998-09-17 |
AU6562298A (en) | 1998-09-29 |
EP1007699A4 (en) | 2001-04-11 |
US6376248B1 (en) | 2002-04-23 |
JP4265699B2 (ja) | 2009-05-20 |
EP2206787A2 (en) | 2010-07-14 |
EP1007699A1 (en) | 2000-06-14 |
US6051429A (en) | 2000-04-18 |
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