OSI-027

製品コードS2624 バッチS262404

印刷

化学情報

Chemical Structure Synonyms ASP4786, CERC 006, AEVI-006 Storage
(From the date of receipt)		 3 years -20°C powder
1 years -80°C in solvent
化学式

C21H22N6O3
分子量 406.44 CAS No. 936890-98-1
Solubility (25°C)* 体外 DMSO 81 mg/mL (199.29 mM)
Water Insoluble
Ethanol Insoluble
体内 （毎回新しく調製した物を用意してください）
Homogeneous suspension
CMC-NA
≥5mg/ml Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

溶剤液（一定の濃度）を調合する

生物活性

製品説明 OSI-027（ASP4786、CERC 006、AEVI-006）は、mTORC1およびmTORC2の選択的かつ強力なデュアル阻害剤であり、セル-フリーアッセイでのIC50はそれぞれ22 nMと65 nMです。PI3Kα、PI3Kβ、PI3Kγ、またはDNA-PKと比較して、mTORに対して100倍以上の選択性が観察されています。OSI-027はがん細胞でAutophagyを誘導します。
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, this compound inhibits mTOR signaling of phospho-4E-BP1 with an IC50 of 1 μM in cell-based assays. It 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. A recent study shows that inhibition of mTORC1/2 by this chemical effectively suppresses phosphorylation of Akt (S473) and cell proliferation in breast cancer cells.

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 this compound potently inhibits tumor growth more than mTORC1 inhibition by rapamycin.

プロトコル(参考用のみ)

キナーゼアッセイ 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.
細胞アッセイ 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 this compound. 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.
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.
投薬量 ≤65 mg/kg
投与方法 Administered via gavage.

参考

  • https://pubmed.ncbi.nlm.nih.gov/21363918/
  • https://pubmed.ncbi.nlm.nih.gov/21415215/
  • https://pubmed.ncbi.nlm.nih.gov/22476852/
  • https://pubmed.ncbi.nlm.nih.gov/21673091/

カスタマーフィードバック

Data from [Urol Oncol, 2013, 1078-1439(13)00251-2]

Data from [Data independently produced by , , Br J Cancer, 2016, 114(6):650-8]

Data from [Data independently produced by , , Cell Physiol Biochem, 2018, 46(2):676-686]

Selleckの高級品が、幾つかの出版された研究調査結果（以下を含む）で使われた：

Changes in Melanoma Cell Morphology Following Inhibition of Cell Invasion by Third-Generation mTOR Kinase Inhibitors [ Int J Mol Sci, 2025, 26(16)7770] PubMed: 40869090
Volatilomic response to targeted cancer therapy in vitro [ Sci Rep, 2025, 15(1):19445] PubMed: 40461777
Three generations of mTOR kinase inhibitors in the activation of the apoptosis process in melanoma cells [ J Cell Commun Signal, 2023, 17(3):975-989] PubMed: 37097377
hnRNP C modulates MERS-CoV and SARS-CoV-2 replication by governing the expression of a subset of circRNAs and cognitive mRNAs [ Emerg Microbes Infect, 2022, 11(1):519-531] PubMed: 35060842
Combined inhibition of BET bromodomain and mTORC1/2 provides therapeutic advantage for rhabdomyosarcoma by switching cell death mechanism [ Mol Carcinog, 2022, 10.1002/mc.23414] PubMed: 35472745
Minimal mitochondrial respiration is required to prevent cell death by inhibition of mTOR signaling in CoQ-deficient cells [ Cell Death Discov, 2021, 7(1):201] PubMed: 34349107
Dual Inhibition of mTORC1/2 Reduces Migration of Cholangiocarcinoma Cells by Regulation of Matrixmetalloproteinases [ Front Cell Dev Biol, 2021, 9:785979] PubMed: 35096817
Mitochondrial Genome-Derived circRNA mc-COX2 Functions as an Oncogene in Chronic Lymphocytic Leukemia [ Mol Ther Nucleic Acids, 2020, 1;20:801-811] PubMed: 32438315
Combined mTORC1/mTORC2 inhibition blocks growth and induces catastrophic macropinocytosis in cancer cells. [ Proc Natl Acad Sci U S A, 2019, 10.1073/pnas.1911393116] PubMed: 31732667
Reverting chemoresistance of targeted agents by a ultrasoluble dendritic nanocapsule. [ J Control Release, 2019, 317:67-77] PubMed: 31756395

長期の保管のために-20°Cの下で製品を保ってください。

人間や獣医の診断であるか治療的な使用のためにでない。

各々の製品のための特定の保管と取扱い情報は、製品データシートの上で示されます。大部分のSelleck製品は、推薦された状況の下で安定です。製品は、推薦された保管温度と異なる温度で、時々出荷されます。長期の保管のために必要とされてそれと異なる温度で、多くの製品は、短期もので安定です。品質を維持するが、夜通しの積荷のために最も経済的な貯蔵状況を用いてあなたの送料を保存する状況の下に、製品が出荷されることを、我々は確実とします。製品の受領と同時に、製品データシートの上で貯蔵推薦に従ってください。