ホームページ PI3K/Akt/mTOR mTOR inhibitor - Autophagy activator OSI-027 For research use only.

OSI-027

別名：ASP4786, CERC 006, AEVI-006

OSI-027 (ASP4786, CERC 006, AEVI-006) is a selective and potent dual inhibitor of mTORC1 and mTORC2 with IC50 of 22 nM and 65 nM in cell-free assays, and more than 100-fold selectivity observed for mTOR than PI3Kα, PI3Kβ, PI3Kγ or DNA-PK. OSI-027 induces autophagy in cancer cells.

OSI-027化学構造

CAS No. 936890-98-1

サイズ 価格(税別) 在庫状況
10mM (1mL in DMSO) JPY 30000 国内在庫あり
JPY 22000 国内在庫あり
JPY 37000 国内在庫あり
JPY 85500 国内在庫あり
JPY 448500 国内在庫なし(納期7~10日)

代表番号: 045-509-1970|電子メール：[email protected]
よく尋ねられる質問

文献中Selleckの製品使用例（38）

製品安全説明書

現在のバッチを見る： 純度： 99.04%
99.04

OSI-027関連製品

シグナル伝達経路

mTOR Signaling Pathways

mTORシグナル伝達経路

mTOR阻害剤の選択性比較

Cell Data

Cell Lines Assay Type Concentration Incubation Time 活性情報 PMID
human SQ20B cells Cytotoxicity assay 72 h Cytotoxicity against human SQ20B cells after 72 hrs by MTT assay, IC50=1.3 μM 24445311
human Caki1 cells Cytotoxicity assay 72 h Cytotoxicity against human Caki1 cells after 72 hrs by MTT assay, IC50=1.9 μM 24445311
human SKHEP1 cells Cytotoxicity assay 72 h Cytotoxicity against human SKHEP1 cells after 72 hrs by MTT assay, IC50=3.2 μM 24445311
human DU145 cells Cytotoxicity assay 72 h Cytotoxicity against human DU145 cells after 72 hrs by MTT assay, IC50=4.1 μM 24445311
human HCT116 cells Cytotoxicity assay 72 h Cytotoxicity against human HCT116 cells after 72 hrs by MTT assay, IC50=5.6 μM 24445311
human ColoR cells Cytotoxicity assay 72 h Cytotoxicity against human ColoR cells after 72 hrs by MTT assay, IC50=5.8 μM 24445311
human COLO205 cells Cytotoxicity assay 72 h Cytotoxicity against human COLO205 cells after 72 hrs by MTT assay, IC50=5.8 μM 24445311
human OVCAR3 cells Cytotoxicity assay 72 h Cytotoxicity against human OVCAR3 cells after 72 hrs by MTT assay, IC50=8.1 μM 24445311
human HOP62 cells Cytotoxicity assay 72 h Cytotoxicity against human HOP62 cells after 72 hrs by MTT assay, IC50=41 μM 24445311
human COLO205 cells Cytotoxicity assay 72 h Cytotoxicity against human COLO205 cells after 72 hrs by MTT assay 24836070
human HOP62 cells Cytotoxicity assay 72 h Cytotoxicity against human HOP62 cells after 72 hrs by MTT assay 24836070
他の多くの細胞株試験データをご覧になる場合はこちらをクリックして下さい

生物活性

製品説明 OSI-027 (ASP4786, CERC 006, AEVI-006) is a selective and potent dual inhibitor of mTORC1 and mTORC2 with IC50 of 22 nM and 65 nM in cell-free assays, and more than 100-fold selectivity observed for mTOR than PI3Kα, PI3Kβ, PI3Kγ or DNA-PK. OSI-027 induces autophagy in cancer cells.
Targets
mTOR [4]
(Cell-free assay)		 mTORC1 [1]
(Cell-free assay)		 mTORC2 [1]
(Cell-free assay)		 PI3Kγ [4]
(Cell-free assay)
4 nM 22 nM 65 nM 0.42 μM
In Vitro
In vitro

OSI-027 shows the selective and ATP competitive inhibition activities against mTORC1 and mTORC2 with IC50 of 22 nM and 65 nM, respectively. In addition, OSI-027 inhibits mTOR signaling of phospho-4E-BP1 with an IC50 of 1 μM in cell-based assays. [1] OSI-027 exhibits anti-proliferative activities against several acute leukemia cell lines of myeloid/megakaryocytic origin in a dose-dependent manner, including U937, KG-1, KBM-3B, ML-1, HL-60, and MEG-01 cells. [2] A recent study shows that inhibition of mTORC1/2 by OSI-027 effectively suppresses phosphorylation of Akt (S473) and cell proliferation in breast cancer cells. [3]
Kinase Assay Biochemical assays
mTORC1 and mTORC2 inhibition is assayed using native enzyme complex immunoprecipitated from HeLa lysates at 1 mM ATP. To prepare whole cell lysates from HeLa cells, 25 g cell pellet is lysed in 60 mL of ice-cold buffer A [40 mM HEPES (pH 7.5), 120 mM NaCl, 1 mM EDTA, 10 mM sodium pyrophosphate, 10 mM glycerophosphate, 50 mM NaF, 0.5 mM orthovanadate, and EDTA-free protease inhibitors containing 0.3% CHAPS] for 30 minutes on a magnetic stirrer in a cold room. After clearing of the lysates by centrifugation at 13,000 g for 10 minutes, Protein G-coated 384-well plates are incubated with 0.25 μg of mTOR antibody in 15 μL of buffer A for 1 hour at 4 °C. To each well, 40 μg of HeLa cell lysate in 15 μL of buffer A is added and incubated overnight at 4 °C to immunoprecipitate mTOR complexes. Plates are washed 3 times with buffer A and twice with immunoprecipitation wash buffer [Buffer B: 50 mM HEPES (pH 7.5) and 150 mM NaCl]. OSI-027 is added at 10 μM concentration to each well and DMSO is added to the control wells. The reaction is started by adding 150 ng of His-tagged 4E-BP1 as a substrate in the presence or absence of 100 μM ATP to each well in 25 μL of freshly prepared kinase buffer [Buffer C: 20 mM HEPES (pH 7.5), 10 mM MgCl2, 4 mM MnCl2, 10 mM β-mercaptoethanol, and 200 μM vanadate] and incubated at room temperature (RT) for 30 minutes. The reaction is stopped by transferring 25 μL of reaction mixture from each well to corresponding wells of fresh Ni-chelate-coated plates and incubated overnight at 4 °C followed by 2 hours at 37 °C. To detect phosphorylation of 4E-BP1, the plates are washed once with TBST (Tris-buffered saline containing 0.1% Tween-20) containing 5% skim milk powder. To each well, 25 μL of 1:1,000 diluted phospho-4E-BP1 antibodies in TBST containing 5% skim milk are added and incubated for 1 hour at RT. The plates are washed once with TBST and then 25 μL of anti-rabbit HRP (diluted 1:10,000) in TBST containing 5% skim milk is added. The plates are incubated for 1 hour at RT and washed 5 times with TBST. For detection of phospho-4E-BP1, 25 μL of chemiluminescent reagents A+B is added and chemiluminescence is measured using an Analyst plate reader.
細胞実験 細胞株 U937, KG-1, KBM-3B, ML-1, HL-60, and MEG-01
濃度 0-10 μM
反応時間 72 hours
実験の流れ

Inhibition of proliferation is measured using the Cell Titer Glo Assay , as noted in figure legends. To generate dose–response curves, cell lines are seeded at a density of 5,000 cells per well in a 96-well plate. After 24 hours of plating, cells are dosed with varying concentrations of either OSI-027 or rapamycin. The signal for Cell Titer Glo Assay is determined 72 hours after dosing and normalized to that of vehicle-treated controls. Inhibition of proliferation, relative to vehicle-treated controls, is expressed as a fraction of 1 and graphed using PRISM software.
実験結果図 Methods Biomarkers 結果図 PMID
Western blot p-mTOR / mTOR / p-AKT / AKT / p-P70S6K / P70S6K / p-4EBP1 / 4EBP1 26026051
Immunofluorescence LC3 24610825
Growth inhibition assay Cell viability 25823922
In Vivo
In Vivo

In GEO colorectal xenograft, OSI-027 (65 mg/kg) inhibits both mTORC1 and mTORC2 effectors, including 4E-BP1, Akt, and S6 phosphorylation. Furthermore, mTORC1 and mTORC2 inhibition together by OSI-027 potently inhibits tumor growth more than mTORC1 inhibition by rapamycin. [1]
動物実験 動物モデル GEO colorectal cells are injected s.c. into the right flank of nu/nu CD-1 mice.
投与量 ≤65 mg/kg
投与経路 Administered via gavage.
NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT00698243 Completed
Any Solid Tumor or Lymphoma
Astellas Pharma Inc
 June 2008 Phase 1

化学情報

分子量 406.44 化学式

C21H22N6O3
CAS No. 936890-98-1 SDF Download OSI-027 SDFをダウンロードする
Smiles COC1=CC=CC2=C1NC(=C2)C3=C4C(=NC=NN4C(=N3)C5CCC(CC5)C(=O)O)N
保管

In vitro
Batch:

DMSO : 81 mg/mL ( (199.29 mM)； 吸湿したDMSOは溶解度を減少させます。新しいDMSOをご使用ください。)

Water : Insoluble

Ethanol : Insoluble

モル濃度計算器

in vivo
Batch:

Add solvents to the product individually and in order.

投与溶液組成計算機

実験計算

モル濃度計算器

質量 濃度 体積 分子量

投与溶液組成計算機(クリア溶液)

ステップ１：実験データを入力してください。（実験操作によるロスを考慮し、動物数を1匹分多くして計算・調製することを推奨します）

mg/kg g μL 匹

ステップ２：投与溶媒の組成を入力してください。（ロット毎に適した溶解組成が異なる場合があります。詳細については弊社までお問い合わせください）

% DMSO % % Tween 80 % ddH2O
%DMSO %

計算結果：

投与溶媒濃度： mg/ml;

DMSOストック溶液調製方法： mg 試薬を μL DMSOに溶解する（濃度 mg/mL, 注：濃度が当該ロットのDMSO溶解度を超える場合はご連絡ください。 )

投与溶媒調製方法：Take μL DMSOストック溶液に μL PEG300，を加え、完全溶解後μL Tween 80，を加えて完全溶解させた後 μL ddH2O，を加え完全に溶解させます。

投与溶媒調製方法：μL DMSOストック溶液に μL Corn oil，を加え、完全溶解。

注意：１.ストック溶液に沈殿、混濁などがないことをご確認ください；
２.順番通りに溶剤を加えてください。次のステップに進む前に溶液に沈殿、混濁などがないことを確認してから加えてください。ボルテックス、ソニケーション、水浴加熱など物理的な方法で溶解を早めることは可能です。

技術サポート

ストックの作り方、阻害剤の保管方法、細胞実験や動物実験の際に注意すべき点など、製品を取扱う時に問い合わせが多かった質問に対しては取扱説明書でお答えしています。

Handling Instructions

他に質問がある場合は、お気軽にお問い合わせください。

* 必須

大学・企業名を記入してください
名前を記入してください
電子メール・アドレスを記入してください 有効なメールアドレスを入力してください
お問い合わせ内容をご入力ください
Tags: OSI-027を買う | OSI-027 ic50 | OSI-027供給者 | OSI-027を購入する | OSI-027費用 | OSI-027生産者 | オーダーOSI-027 | OSI-027化学構造 | OSI-027分子量 | OSI-027代理店