OSI-027

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.

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
5mg	JPY 22000	国内在庫あり
10mg	JPY 37000	国内在庫あり
50mg	JPY 85500	国内在庫あり
1g	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

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製品安全説明書

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OSI-027関連製品

関連ターゲット	mTORC1 mTORC2	もっと見る
関連製品	Rapamycin (Sirolimus) Everolimus AZD8055 Torin 1 Ridaforolimus (Deforolimus, MK-8669) Sapanisertib (MLN0128) Temsirolimus Torkinib (PP242) Vistusertib (AZD2014) MHY1485 KU-0063794 Torin 2 WYE-354 WYE-125132 (WYE-132) 3BDO Palomid 529 (P529) Zotarolimus (ABT-578) Salidroside GDC-0349 WAY-600 Onatasertib (CC 223) XL388 WYE-687	もっと見る
関連化合物ライブラリー	Kinase Inhibitor Library PI3K/Akt Inhibitor Library Apoptosis Compound Library Cell Cycle compound library NF-κB Signaling Compound Library	もっと見る

シグナル伝達経路

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	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
Kinase Assay	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
臨床試験 (data from https://clinicaltrials.gov, updated on 2024-01-11)
NCT00698243	Completed	Any Solid Tumor or Lymphoma	Astellas Pharma Inc	June 2008	Phase 1

参考
[1] Falcon BL, et al. Cancer Res, 2011, 71(5), 1573-1583. [2] Altman JK, et al. Clin Cancer Res, 2011, 17(13), 4378-4388. [3] Li H, et al. Breast Cancer Res Treat, 2012, 134(3), 1057-1066. [4] Bhagwat SV, et al. Mol Cancer Ther. 2011, 10(8), 1394-1406. + 展開

参考

[4] Bhagwat SV, et al. Mol Cancer Ther. 2011, 10(8), 1394-1406.

+ 展開

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	3年-20°C粉	溶液の保管冷凍と解凍の繰り返しを避けるため小分けにしてください	1年-80°Cin solvent
保管	3年-20°C粉	溶液の保管冷凍と解凍の繰り返しを避けるため小分けにしてください	1个月-20°Cin solvent

In vitro
Batch:

DMSO : 18 mg/mL ( (44.28 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
=	×	×

希釈計算器分子量計算器

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）

投与量 mg/kg 動物平均体重 g 投与体積（動物毎）μL 動物数匹

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。

C1=C0/X	C1:	LOG(C1):
C2=C1/X	C2:	LOG(C2):
C3=C2/X	C3:	LOG(C3):
C4=C3/X	C4:	LOG(C4):
C5=C4/X	C5:	LOG(C5):
C6=C5/X	C6:	LOG(C6):
C7=C6/X	C7:	LOG(C7):
C8=C7/X	C8:	LOG(C8):