OSI-027

Catalog No.S2624 Synonyms: ASP4786, CERC 006, AEVI-006

For research use only.

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. Phase 1.

OSI-027 Chemical Structure

CAS No. 936890-98-1

Selleck's OSI-027 has been cited by 37 publications

Purity & Quality Control

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Biological Activity

Description 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. Phase 1.
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

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]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
human SQ20B cells MVPDfZRwfG:6aXPpeJkh[XO|YYm= MkD2O|IhcA>? NUjJXXZ5S3m2b4TvfIlkcXS7IHHnZYlve3RiaIXtZY4hW1F{MFKgZ4VtdHNiYX\0[ZIhPzJiaILzJIJ6KE2WVDDhd5NigSxiSVO1NF0yNjNizszN MUSyOFQ1PTNzMR?=
human Caki1 cells NFi2[5lEgXSxdH;4bYNqfHliYYPzZZk> M3LSRlczKGh? Mm\FR5l1d3SxeHnjbZR6KGGpYXnud5QhcHWvYX6gR4FscTFiY3XscJMh[W[2ZYKgO|IhcHK|IHL5JG1VXCCjc4PhfUwhUUN3ME2xMlkh|ryP MWiyOFQ1PTNzMR?=
human SKHEP1 cells NV7vUJhyS3m2b4TvfIlkcXS7IHHzd4F6 M1v5V|czKGh? MnXiR5l1d3SxeHnjbZR6KGGpYXnud5QhcHWvYX6gV2tJTVBzIHPlcIx{KGGodHXyJFczKGi{czDifUBOXFRiYYPzZZktKEmFNUC9N{4zKM7:TR?= NIewcoQzPDR2NUOxNS=>
human DU145 cells MXLDfZRwfG:6aXPpeJkh[XO|YYm= MVW3NkBp Ml[xR5l1d3SxeHnjbZR6KGGpYXnud5QhcHWvYX6gSHUyPDViY3XscJMh[W[2ZYKgO|IhcHK|IHL5JG1VXCCjc4PhfUwhUUN3ME20MlEh|ryP NV;iZ3o2OjR2NEWzNVE>
human HCT116 cells M1vrUGN6fG:2b4jpZ4l1gSCjc4PhfS=> M1;yOlczKGh? MlO4R5l1d3SxeHnjbZR6KGGpYXnud5QhcHWvYX6gTGNVOTF4IHPlcIx{KGGodHXyJFczKGi{czDifUBOXFRiYYPzZZktKEmFNUC9OU43KM7:TR?= NUj1fYJnOjR2NEWzNVE>
human ColoR cells M{j6OGN6fG:2b4jpZ4l1gSCjc4PhfS=> M17ocFczKGh? MnfhR5l1d3SxeHnjbZR6KGGpYXnud5QhcHWvYX6gR49td1JiY3XscJMh[W[2ZYKgO|IhcHK|IHL5JG1VXCCjc4PhfUwhUUN3ME21Mlgh|ryP NGe0O3IzPDR2NUOxNS=>
human COLO205 cells NUjURnllS3m2b4TvfIlkcXS7IHHzd4F6 MWC3NkBp NYDFPG9[S3m2b4TvfIlkcXS7IHHnZYlve3RiaIXtZY4hS0:OT{KwOUBk\WyuczDh[pRmeiB5MjDodpMh[nliTWTUJIF{e2G7LDDJR|UxRTVwODFOwG0> Mli0NlQ1PDV|MUG=
human OVCAR3 cells MmDWR5l1d3SxeHnjbZR6KGG|c3H5 MlHxO|IhcA>? M4XzV2N6fG:2b4jpZ4l1gSCjZ3HpcpN1KGi3bXHuJG9XS0GUMzDj[YxteyCjZoTldkA4OiCqcoOgZpkhVVSWIHHzd4F6NCCLQ{WwQVgvOSEQvF2= NIT6S|czPDR2NUOxNS=>
human HOP62 cells NUmzbGZJS3m2b4TvfIlkcXS7IHHzd4F6 M1XXb|czKGh? M2PpN2N6fG:2b4jpZ4l1gSCjZ3HpcpN1KGi3bXHuJGhQWDZ{IHPlcIx{KGGodHXyJFczKGi{czDifUBOXFRiYYPzZZktKEmFNUC9OFEh|ryP M2H3SVI1PDR3M{Gx
human COLO205 cells NEXwTWNEgXSxdH;4bYNqfHliYYPzZZk> NUTlfmdCPzJiaB?= NGrhOJpEgXSxdH;4bYNqfHliYXfhbY5{fCCqdX3hckBEV0yRMkC1JINmdGy|IHHmeIVzKDd{IHjyd{BjgSCPVGSgZZN{[Xl? NHjldXQzPDh|NkC3NC=>
human HOP62 cells NFrCRYdEgXSxdH;4bYNqfHliYYPzZZk> MV:3NkBp M13sT2N6fG:2b4jpZ4l1gSCjZ3HpcpN1KGi3bXHuJGhQWDZ{IHPlcIx{KGGodHXyJFczKGi{czDifUBOXFRiYYPzZZk> NGXRW4EzPDh|NkC3NC=>
Assay
Methods Test Index 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 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]

Protocol (from reference)

Kinase Assay:

[1]

  • 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.

Cell Research:

[2]

  • Cell lines: U937, KG-1, KBM-3B, ML-1, HL-60, and MEG-01
  • Concentrations: 0-10 μM
  • Incubation Time: 72 hours
  • Method:

    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.

Animal Research:

[1]

  • Animal Models: GEO colorectal cells are injected s.c. into the right flank of nu/nu CD-1 mice.
  • Dosages: ≤65 mg/kg
  • Administration: Administered via gavage.

Solubility (25°C)

In vitro

In vivo

Add solvents to the product individually and in order
(Data is from Selleck tests instead of citations):
1% DMSO+30% polyethylene glycol+1% Tween 80
For best results, use promptly after mixing.

30 mg/mL

Chemical Information

Molecular Weight 406.44
Formula

C21H22N6O3

CAS No. 936890-98-1
Storage 3 years -20°C powder
2 years -80°C in solvent
Smiles COC1=CC=CC2=C1NC(=C2)C3=C4C(=NC=NN4C(=N3)C5CCC(CC5)C(=O)O)N

In vivo Formulation Calculator (Clear solution)

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Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
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Molarity Calculator

Mass Concentration Volume Molecular Weight

Clinical Trial Information

NCT Number Recruitment Interventions Conditions Sponsor/Collaborators Start Date Phases
NCT00698243 Completed Drug: OSI-027 Any Solid Tumor or Lymphoma Astellas Pharma Inc June 2008 Phase 1

(data from https://clinicaltrials.gov, updated on 2022-01-17)

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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