WO2010024262A1 - トランスフェクション剤 - Google Patents
トランスフェクション剤 Download PDFInfo
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- WO2010024262A1 WO2010024262A1 PCT/JP2009/064814 JP2009064814W WO2010024262A1 WO 2010024262 A1 WO2010024262 A1 WO 2010024262A1 JP 2009064814 W JP2009064814 W JP 2009064814W WO 2010024262 A1 WO2010024262 A1 WO 2010024262A1
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- sirna
- peptide surfactant
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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
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
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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
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
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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
- 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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- 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
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
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- the present invention relates to a transfection agent containing a peptide surfactant.
- RNAi can suppress the expression of a specific gene, it is expected to be applied to the treatment of diseases.
- the transfection agent used when transfection of siRNA into cells is mainly cationic phospholipid, and its toxicity has been pointed out.
- transfection agents are desired to have an extremely low toxicity, and development of transfection agents with lower toxicity is awaited.
- Peptide surfactants are known to have the action of stabilizing membrane proteins such as the G protein-coupled receptor bovine rhodopsin and the property of self-organization (Non-patent Documents 1 and 2).
- Non-patent Documents 1 and 2 Non-patent Documents 1 and 2.
- its use as a transfection agent has not been known.
- X Zhao et al., PNAS, Vol.103, No. 47, 17707-17712 A Nagai et al., J. Nanosci. Nanotechnol. Vol. 7, No. 7, 1-7
- the problem to be solved by the present invention is to provide a clinically applicable transfection agent with lower toxicity.
- a peptide surfactant is useful as a transfection agent, and has completed the present invention.
- a transfection agent comprising a peptide surfactant, 2.
- a pharmaceutical comprising the transfection agent according to any one of 1 to 3 above and a nucleic acid, It is.
- transduced into 3 types of human malignant glioma culture cells by the peptide surfactant is confirmation of the RNA interference effect by siRNA introduce
- transduced into 3 types of human malignant glioma culture cells by the peptide surfactant is confirmation of the RNA interference effect by siRNA introduce
- a peptide surfactant includes 6 to 10 amino acid residues, and has a length of about 2 to 3 nm.
- Conventional surfactants such as n-dodecyl- ⁇ It exhibits the same characteristics as -D-maltoside (DM) and octyl-D-glucoside (OG).
- the peptide surfactant of the present invention preferably comprises a head composed of 4 to 10 hydrophilic amino acids and a tail composed of 1 or 2 hydrophobic amino acids.
- GGGGDD G4D2
- GGGGGGDD G6D2
- GGGGGGGGDD G8D2
- GGGGGGGGGDD G10D2
- AAAAAAD A6D
- A6AA A6AA, A6A, A6A, A6A, A6A , VVVVVDD (V6D2), VVVVVVK (V6K), VVVVVVKK (V6K2), LLLLLLD (L6D), LLLLLLDDD (L6D2), LLLLLLLK (L6K), or LLLLLALK6 (A6)
- AAAAAAK A6K
- AAAAAAAAK AAAAAAK
- AAAAAAAAK A6K
- AAAAAAAAK A6K
- the peptide surfactant of the present invention can be produced using a conventionally known solid phase synthesis method.
- Such an artificially synthesized peptide does not contain a substance derived from a living body and can be preferably used because there is no concern about infectivity.
- the term “transfection agent” refers to a composition that can be used to introduce a polynucleotide or gene such as siRNA into a cell and increase its introduction efficiency.
- the transfection agent of the present invention can be blended with pharmaceuticals together with desired nucleic acids.
- the pharmaceutical product of the present invention can be used for either oral administration or parenteral administration.
- the nucleic acid used in the pharmaceutical product of the present invention is a part of the base sequence of a gene to be used as a target for treatment or the like, and is composed of DNA or RNA nucleic acid. A gene that can control the function of a gene. Such nucleic acids are generally known as nucleic acid drugs.
- a decoy nucleic acid medicine a drug that inhibits gene expression by binding a transcriptional regulator to an oligonucleotide with the same sequence as a certain region (cis element) of DNA in the nucleus to which the transcriptional regulator binds
- Nucleic acid aptamers that can specifically bind to other molecules, ribozymes that have catalytic activity to cleave RNAs, and antisense RNAs or target genes that inhibit gene expression by complementary binding to specific regions of the target gene mRNA Examples include siRNA that degrades mRNA by binding to a specific region of mRNA together with an RNA silencing complex.
- Decoy nucleic acid drugs, nucleic acid aptamers, and ribozymes are highly specific nucleic acids that have a three-dimensional structure.
- Antisense RNA is a single-stranded RNA oligomer that has a part of the reverse side of the mRNA sequence, and siRNA is the target.
- These nucleic acids can be artificially produced by a chemical reaction using a nucleic acid synthesizer, or can be produced from cultured cells using a vector.
- biopharmaceuticals consisting of chemical substances in living organisms are attracting attention in terms of their target specificity and biological safety.
- One of the biopharmaceuticals includes nucleic acid drugs, which are currently being developed by many research institutions and pharmaceutical companies.
- nucleic acid drugs siRNA-specific gene suppression effects have attracted attention in recent years, and siRNA drugs for various diseases are being developed.
- An example of a therapeutic effect expected by using siRNA is treatment of glioblastoma, which is a kind of brain tumor. In glioblastoma, it is known that RNAi targeting MMP-2 induces apoptosis of tumor cells and suppresses tumor growth.
- Pharmaceutical forms include tablets, capsules, granules, syrups and the like. These pharmaceuticals can be produced using additives in normal pharmaceutical production.
- the dosage of the pharmaceutical product of the present invention is not particularly limited.
- it is 0.001 to 10 mg / kg body weight per day.
- the above dose can be administered once a day or divided into 2 to 3 times a day, and can be appropriately increased or decreased depending on the age, disease state, and symptoms.
- the additive examples include lactose, dextrin, sucrose, mannitol, corn starch, sorbitol, crystalline cellulose, polyvinylpyrrolidone and the like, and these can be used alone or in appropriate combination.
- These pharmaceuticals can be produced by a method suitable for each pharmaceutical form according to the description of the Japanese Pharmacopoeia.
- a flavoring agent, a coloring agent, a sweetener, etc. can also be used suitably.
- the content of these additives can be appropriately selected by those skilled in the art.
- quasi drugs examples include tablets, capsules, granules, jellies, and drinks. These quasi-drugs can be manufactured using the additives in normal quasi-drug manufacture. Furthermore, these quasi drugs may contain other active ingredients such as vitamins. In addition, additives such as sweeteners, flavoring agents, coloring agents, and antioxidants may be used alone or in appropriate combination. These quasi drugs can be produced by methods well known to those skilled in the art.
- Cytotoxicity evaluation was performed using cultured human cells for two types of peptide surfactant and cationic liposome gene transfer agent that are currently widely used.
- Peptide surfactant (sequence: Ac-AAAAAAK-NH 2 , manufactured by Celtek) 2. Lipofectamine 2000 (Invitrogen) 3. siFECTOR (B-Bridge International, Inc.) ⁇ Cell culture of human malignant glioma (U87MG, purchased from ATCC) ⁇ MTT assay reagent (quantification of cell proliferation number) CellTiter 96 Aqueous One Solution Cell Proliferation Assay Kit (Promega)
- ⁇ Method> Cells were seeded in 96-well plates (cell density: 2 ⁇ 10 4 / well) according to ATCC recommended protocol and incubated overnight. Replaced with fresh medium and added additive (final concentrations: 0.0005, 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 25, 50, 75, 100, 250, 500 ⁇ g / ml) . Cells were cultured for 24 hours. Thereafter, CellTiter 96 Aqueous One Solution was added (20 ⁇ l / well), and after 4 hours, the absorbance at 495 nm was measured with a plate reader, and the number of viable cells was quantified.
- the toxicity of the peptide surfactant was the lowest.
- the LD50 was 285.9 ⁇ g / ml peptide surfactant, 42.49 ⁇ g / ml Lipofectamine 2000, and 25.3 ⁇ g / ml siFECTOR.
- the peptide surfactant showed no cytotoxicity at 25 ⁇ g / ml or less.
- ⁇ Material> • Peptide surfactant sequence: Ac-AAAAAAK-NH 2 , manufactured by Celtek • siRNA siRNA / MMP-2, manufactured by Santa Cruz, siRNA fluorescent labeling reagent Label IT Cy-3 Labeling Kit, manufactured by Mirus, 3 types of cultured human malignant glioma cells (U87MG, T98G, U251MG, purchased from ATCC, respectively) ⁇ Method> Cy3 was labeled with siRNA fluorescent labeling reagent on siRNA / MMP-2 according to Mirus protocol. According to the ATCC protocol, 5000 cells were seeded on a chambered slide glass and placed overnight.
- the medium was removed, and added to 1 ml of the medium so that the final concentration was 50 nM of siRNA and 20 ⁇ g / ml of peptide surfactant, followed by culturing for 8 hours.
- siRNA was cultured in 1 ml of medium so that the final concentration was 50 nM.
- fluorescence observation with a Cy3 filter and observation with clear vision were performed.
- Time-dependent analysis of the amount of siRNA introduced into cells by peptide surfactant The time-dependent change in the amount of siRNA introduced into cells by the peptide surfactant was examined by observing fluorescence-labeled siRNA under a microscope.
- Peptide surfactant sequence Ac-AAAAAAK-NH 2 , manufactured by Celtek • Peptide surfactant concentration 0, 0.5, 1.0, 5.0, 10.0, 20.0, 25.0 ⁇ g / mL ⁇ SiRNA siRNA / MMP-2, manufactured by Santa Cruz, siRNA fluorescent labeling reagent Label IT Cy-3 Labeling Kit, manufactured by Mirus, 3 types of cultured human malignant glioma cells (purchased from U87MG, T98G, U251MG, ATCC)
- Cy3 was labeled with siRNA fluorescent labeling reagent on siRNA / MMP-2 according to the standard protocol of Miras. Labeled siRNA was mixed with a peptide surfactant. For 1 ⁇ 10 5 cells seeded in a 24-well plate containing 1 ml / well of medium, the final concentration was 50 nM for siRNA and 20 ⁇ g / ml for peptide surfactant. Incubated for 2, 4, 8, 16, and 24 hours. After washing the wells with PBS, the cells were collected and transferred to a 96-well plate, and the fluorescence was measured.
- Peptide surfactant sequence Ac-AAAAAAK-NH 2 , manufactured by Celtek • Peptide surfactant concentration 0, 0.5, 1.0, 5.0, 10.0, 20.0, 25.0 ⁇ g / mL ⁇ SiRNA siRNA / MMP-2, manufactured by Santa Cruz, siRNA fluorescent labeling reagent Label IT Cy-3 Labeling Kit, manufactured by Mirus, 3 types of cultured human malignant glioma cells (purchased from U87MG, T98G, U251MG, ATCC)
- Cy3 was labeled with siRNA fluorescent labeling reagent on siRNA / MMP-2 according to the standard protocol of Miras. Labeled siRNA was mixed with a peptide surfactant. For 1 ⁇ 10 5 cells seeded in a 24-well plate containing 1 ml / well of medium, the final concentration is 50 nM for siRNA, 0, 0.5, 1, 5, 10, 20, 25 ⁇ g / g for peptide surfactant. The solution was administered in a volume of ml and cultured for 8 hours. After washing the wells with PBS, the cells were collected and transferred to a 96-well plate, and the fluorescence was measured.
- Peptide surfactant sequence Ac-AAAAAAK-NH 2 , manufactured by Celtek ⁇ Three types of cell human malignant glioma cultured cells (purchased from U87MG, T98G, U251MG, ATCC)
- ⁇ Method> According to the standard protocol of ATCC, 1 ⁇ 10 5 cells were seeded in a 24-well plate and cultured overnight. After removing the medium, the following solution was added to the same culture conditions as described above so that the final concentration was as follows, followed by culturing for 8 hours.
- siRNA 50 nM / peptide surfactant 20 ⁇ g / ml The siRNA was reverse transcribed into cDNA using Taqman Gene Expression Cell-to-Ct Kit (Applied Biosystems). The gene expression assay (Applied Biosystems) targeted MMP-2 and beta actin as an internal standard, and quantitative analysis was performed using an Applied Biosystems 7500 real-time PCR system.
- ⁇ MMP-2 mRNA expression was significantly suppressed only when siRNA 50 nM / peptide surfactant 20 ⁇ g / ml was administered.
- -Since siRNA alone has no effect of introducing into cells, it was considered that siRNA was incorporated into cells by the effect of siRNA introduction of the peptide surfactant, resulting in an RNA interference effect.
- ⁇ Suppression of MMP-2 gene is not observed in a mixture of scrambled oligo RNA and peptide surfactant that has no homology in the human genome and has no specificity for the gene. It was thought that this was not a non-specific reaction that did not depend on.
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Abstract
Description
X Zhao et al., PNAS, Vol.103, No. 47, 17707-17712 A Nagai et al., J. Nanosci. Nanotechnol. Vol. 7, No.7, 1-7
1.ペプチド界面活性剤を含むトランスフェクション剤、
2.ペプチド界面活性剤が、4~10の親水性アミノ酸からなる頭部及び1又は2の疎水性アミノ酸からなる尾部からなる、上記1に記載のトランスフェクション剤、
3.ペプチド界面活性剤が、AAAAAAD又はAAAAAAKである、上記1に記載のトランスフェクション剤、
4.上記1~3のいずれか1に記載のトランスフェクション剤及び核酸を含む、医薬品、
である。
ペプチド界面活性剤と現在広く使用されているカチオン性リポソーム遺伝子導入剤2種について、ヒト培養細胞を用いて細胞毒性評価を行った。
・添加物質
1.ペプチド界面活性剤(配列:Ac-AAAAAAK-NH2、Celtek社製)
2.リポフェクトアミン2000(インビトロジェン社製)
3.siFECTOR (B-Bridge International, Inc.社製)
・細胞
ヒト悪性グリオーマ培養細胞(U87MG、ATCCから購入)
・MTTアッセイ試薬(細胞増殖数定量)
CellTiter 96 Aqueous One Solution Cell Proliferation Assayキット(プロメガ社製)
ATCC推奨プロトコルに従って、細胞を96ウェルプレートに播種し(細胞密度:2×104/well)、一晩インキュベーションした。新鮮な培地に交換し、添加物質を添加した(最終濃度:0.0005, 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 25, 50, 75, 100, 250, 500μg/ml)。細胞を24時間培養した。その後、CellTiter 96 Aqueous One Solutionを添加(20μl/well)し、4時間後、プレートリーダーで495nmの吸光光度を測定し、生存細胞数の定量を行った。
ペプチド界面活性剤によってsiRNAが細胞に導入されたことを、蛍光標識したsiRNAを顕微鏡観察することで確認した。
・ペプチド界面活性剤
配列:Ac-AAAAAAK-NH2、Celtek社製
・siRNA
siRNA/MMP-2、Santa Cruz社製
・siRNA蛍光標識試薬
Label IT Cy-3 Labelling Kit、Mirus社製
・細胞
ヒト悪性グリオーマ培養細胞3種(U87MG、T98G、U251MG、それぞれATCCから購入)
<方法>
Mirus社プロトコルに従い、siRNA/MMP-2にsiRNA蛍光標識試薬でCy3を標識した。ATCCのプロトコルに従い、培養チャンバー付きスライドグラス(Chambered slide glass)に5000個の細胞を播種し一晩置いた。培地を除去し、最終濃度がsiRNA 50nM、ペプチド界面活性剤20μg/mlとなるよう、培地1mlに加え、8時間培養を行った。対照群は、siRNAのみを最終濃度が50nMとなるよう、培地1mlで培養を行った。倒立蛍光顕微鏡で、Cy3用のフィルターでの蛍光観察と明視での観察を行った。
3種の細胞すべてにおいて、ペプチド界面活性剤とsiRNAの両方を加えた試験群でCy3標識したsiRNA の取り込みが確認された。siRNAのみを加えた対照群では、いずれの細胞においても標識されたsiRNAの取り込みは認められなかった。
ペプチド界面活性剤による細胞へのsiRNA導入量の経時的変化を、蛍光標識したsiRNAを顕微鏡観察することで検討した。
・ペプチド界面活性剤
配列:Ac-AAAAAAK-NH2、Celtek社製
・ペプチド界面活性剤濃度
0、0.5、1.0、5.0、10.0、20.0、25.0μg/mL
・siRNA
siRNA/MMP-2、Santa Cruz社製
・siRNA蛍光標識試薬
Label IT Cy-3 Labelling Kit、Mirus社製
・細胞
ヒト悪性グリオーマ培養細胞3種(U87MG、T98G、U251MG、ATCCから購入)
Mirus社標準プロトコルに従い、siRNA/MMP-2にsiRNA蛍光標識試薬でCy3を標識した。ラベルしたsiRNAをペプチド界面活性剤と混合した。培地が1ml/well入った24wellプレートに播種した1×105個の細胞に対して、最終濃度がsiRNAは50nM、ペプチド界面活性剤は20μg/mlになるよう投与して、0.5、1、2、4、8、16及び24時間培養した。PBSでウェルを洗浄後、細胞を回収して96ウェルプレートに移して蛍光光度を測定した。
・培養1~8時間では、相対蛍光単位(RFU)がほぼ比例的に増加し、時間に比例してsiRNAが細胞に取り込まれることがわかった。
・培養8時間以降は、ほとんどプラトーに達していた。
siRNAの細胞への導入効率がペプチド界面活性剤濃度にどのように依存するかを、蛍光標識したsiRNAを顕微鏡観察することで検討した。
・ペプチド界面活性剤
配列:Ac-AAAAAAK-NH2、Celtek社製
・ペプチド界面活性剤濃度
0、0.5、1.0、5.0、10.0、20.0、25.0μg/mL
・siRNA
siRNA/MMP-2、Santa Cruz社製
・siRNA蛍光標識試薬
Label IT Cy-3 Labelling Kit、Mirus社製
・細胞
ヒト悪性グリオーマ培養細胞3種(U87MG、T98G、U251MG、ATCCから購入)
Mirus社標準プロトコルに従い、siRNA/MMP-2にsiRNA蛍光標識試薬でCy3を標識した。ラベルしたsiRNA をペプチド界面活性剤と混合した。培地が1ml/well入った24ウェルプレートに播種した1×105個の細胞に対して、最終濃度がsiRNAは50nM、ペプチド界面活性剤は0、0.5、1、5、10、20、25μg/mlになるよう投与して、8時間培養した。PBSでウェルを洗浄後、細胞を回収して96ウェルプレートに移して蛍光光度を測定した。
・Y98Gにおいて10μg/ml以上で濃度依存的なRFUの上昇が弱まったものの、3種すべての細胞で濃度の増加に比例したRFUの増加が認められ、細胞へのsiRNAの導入がペプチド界面活性剤の濃度に依存することが明らかとなった。
・実施例3の結果とあわせ、ペプチド界面活性剤を濃度が20μg/mlで8時間細胞にさらすことで、十分有意にsiRNAが細胞に導入されることが明らかになった。
ペプチド界面活性剤によって細胞に導入されたsiRNAがRNA干渉能力を持つことをRT-PCR法で確認を行った。
・ペプチド界面活性剤
配列:Ac-AAAAAAK-NH2、Celtek社製
・細胞
ヒト悪性グリオーマ培養細胞3種(U87MG、T98G、U251MG、ATCCから購入)
ATCC社の標準プロトコルに従って、24ウェルプレートに細胞を1×105個播種し一晩培養した。培地を除去後、前記と同じ培養条件のところへ、最終濃度が以下になるよう次の溶液を入れて8時間培養した。
(1)コントロール(培地)
(2)スクランブルオリゴsiRNA(Dharmacon社製)/ペプチド界面活性剤20μg/ml
(3)siRNA50nM
(4)ペプチド界面活性剤20μg/ml
(5)siRNA 50nM/ペプチド界面活性剤20μg/ml
Taqman Gene Expression Cell-to-Ct Kit (アプライドバイオシステムズ社製)を用いてsiRNAをcDNAに逆転写した。Gene Expression Assay(アプライドバイオシステムズ社製)でMMP-2を標的とし、ベータアクチンを内部標準としてApplied Biosystems 7500リアルタイムPCRシステムによって定量解析を行った。
・siRNA 50nM/ペプチド界面活性剤20μg/mlを投与したもののみで有意にMMP-2 mRNAの発現が抑制されていた。
・siRNAのみでは細胞への導入効果がないことから、ペプチド界面活性剤のsiRNA導入効果によってsiRNAが細胞内へ取り込まれ、その結果RNA干渉効果が生じたと考えられた。
・ヒトゲノム中で相同性が認められない配列で構成され遺伝子に対する特異性のないスクランブルオリゴRNAとペプチド界面活性剤を混合したものでは、MMP-2遺伝子の抑制がみらないことから、RNAの配列に依存しない非特異的反応ではないと考えられた。
Claims (4)
- ペプチド界面活性剤を含むトランスフェクション剤。
- ペプチド界面活性剤が、4~10の親水性アミノ酸からなる頭部及び1又は2の疎水性アミノ酸からなる尾部からなる、請求項1に記載のトランスフェクション剤。
- ペプチド界面活性剤が、AAAAAAD又はAAAAAAKである、請求項1に記載のトランスフェクション剤。
- 請求項1~3のいずれか1項に記載のトランスフェクション剤及び核酸を含む、医薬品。
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JP2010526725A JP5606318B2 (ja) | 2008-08-27 | 2009-08-26 | トランスフェクション剤 |
EP09809913.8A EP2322608B1 (en) | 2008-08-27 | 2009-08-26 | Transfection agent |
ES09809913.8T ES2491565T3 (es) | 2008-08-27 | 2009-08-26 | Agente de transfección |
US13/058,880 US9133484B2 (en) | 2008-08-27 | 2009-08-26 | Transfection agent |
CN2009801334914A CN102131926A (zh) | 2008-08-27 | 2009-08-26 | 转染剂 |
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EP (1) | EP2322608B1 (ja) |
JP (1) | JP5606318B2 (ja) |
KR (1) | KR101647521B1 (ja) |
CN (1) | CN102131926A (ja) |
ES (1) | ES2491565T3 (ja) |
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Cited By (5)
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WO2012023345A1 (ja) | 2010-08-20 | 2012-02-23 | 独立行政法人 国立がん研究センター | がん幹細胞を含むまたはそれに由来するがんの治療、予防および診断のための方法および組成物 |
JP2015519374A (ja) * | 2012-05-31 | 2015-07-09 | マサチューセッツ インスティチュート オブ テクノロジー | ペプチド界面活性剤を使用した分子の持続送達およびその使用 |
WO2018097335A1 (ja) * | 2016-11-25 | 2018-05-31 | 国立大学法人 岡山大学 | ホウ素中性子捕捉療法のための新規bsh複合体 |
WO2019093308A1 (ja) | 2017-11-09 | 2019-05-16 | 国立大学法人広島大学 | miRNAを含むがん治療用医薬組成物 |
US11324703B2 (en) | 2017-12-15 | 2022-05-10 | 3-D Matrix, Ltd. | Surfactant peptide nanostructures and uses thereof in drug delivery |
Citations (1)
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EP0874910A4 (en) | 1995-06-07 | 1999-04-21 | Life Technologies Inc | ENHANCED CATIONIC LIPID TRANSFECTION BY PEPTIDES |
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- 2009-08-26 US US13/058,880 patent/US9133484B2/en active Active
- 2009-08-26 EP EP09809913.8A patent/EP2322608B1/en active Active
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- 2009-08-26 KR KR1020117006882A patent/KR101647521B1/ko active IP Right Grant
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JP2001517939A (ja) * | 1997-03-14 | 2001-10-09 | ライフ テクノロジーズ,インコーポレイテッド | ペプチドによって増強されるトランスフェクション |
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WO2012023345A1 (ja) | 2010-08-20 | 2012-02-23 | 独立行政法人 国立がん研究センター | がん幹細胞を含むまたはそれに由来するがんの治療、予防および診断のための方法および組成物 |
US20130236891A1 (en) * | 2010-08-20 | 2013-09-12 | 3-D Matrix, Ltd. | Method and composition for the treatment, prevention, and diagnosis of cancer containing or derived from cancer stem cells |
US10337012B2 (en) | 2010-08-20 | 2019-07-02 | 3-D Matrix, Ltd. | Method and composition for the treatment, prevention, and diagnosis of cancer containing or derived from cancer stem cells |
JP2015519374A (ja) * | 2012-05-31 | 2015-07-09 | マサチューセッツ インスティチュート オブ テクノロジー | ペプチド界面活性剤を使用した分子の持続送達およびその使用 |
WO2018097335A1 (ja) * | 2016-11-25 | 2018-05-31 | 国立大学法人 岡山大学 | ホウ素中性子捕捉療法のための新規bsh複合体 |
JPWO2018097335A1 (ja) * | 2016-11-25 | 2019-04-18 | 国立大学法人 岡山大学 | ホウ素中性子捕捉療法のための新規bsh複合体 |
WO2019093308A1 (ja) | 2017-11-09 | 2019-05-16 | 国立大学法人広島大学 | miRNAを含むがん治療用医薬組成物 |
EP4265725A2 (en) | 2017-11-09 | 2023-10-25 | Purmx Therapeutics, Inc. | Pharmaceutical composition comprising mirna-3140 for use in treating cancer |
EP4276184A2 (en) | 2017-11-09 | 2023-11-15 | Purmx Therapeutics, Inc. | Pharmaceutical composition comprising mirna-3140 for use in treating cancer |
EP4276183A2 (en) | 2017-11-09 | 2023-11-15 | Purmx Therapeutics, Inc. | Pharmaceutical composition comprising mirna-3140 for use in treating cancer |
US11324703B2 (en) | 2017-12-15 | 2022-05-10 | 3-D Matrix, Ltd. | Surfactant peptide nanostructures and uses thereof in drug delivery |
Also Published As
Publication number | Publication date |
---|---|
US9133484B2 (en) | 2015-09-15 |
JP5606318B2 (ja) | 2014-10-15 |
KR20110055693A (ko) | 2011-05-25 |
EP2322608A4 (en) | 2012-03-21 |
KR101647521B1 (ko) | 2016-08-10 |
EP2322608A1 (en) | 2011-05-18 |
CN102131926A (zh) | 2011-07-20 |
SG193822A1 (en) | 2013-10-30 |
JPWO2010024262A1 (ja) | 2012-01-26 |
US20110152203A1 (en) | 2011-06-23 |
ES2491565T3 (es) | 2014-09-08 |
EP2322608B1 (en) | 2014-05-21 |
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