AZD8055

Catalog No.S1555

AZD8055 is a novel ATP-competitive mTOR inhibitor with IC50 of 0.8 nM in MDA-MB-468 cells with excellent selectivity (∼1,000-fold) against PI3K isoforms and ATM/DNA-PK. Phase 1.

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AZD8055 Chemical Structure

AZD8055 Chemical Structure
Molecular Weight: 465.54

Validation & Quality Control

Cited by 36 publications:

7 customer reviews :

Quality Control & MSDS

Related Compound Libraries

mTOR Inhibitors with Unique Features

Product Information

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  • Research Area
  • Inhibition Profile

Product Description

Biological Activity

Description AZD8055 is a novel ATP-competitive mTOR inhibitor with IC50 of 0.8 nM in MDA-MB-468 cells with excellent selectivity (∼1,000-fold) against PI3K isoforms and ATM/DNA-PK. Phase 1.
Targets mTOR (truncated) [1]
(MDA-MB-468 cells)
mTOR (full length) [1]
(MDA-MB-468 cells)
DNA-PK [1]
(MDA-MB-468 cells)
PI3Kδ [1]
(MDA-MB-468 cells)

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IC50 0.13 nM 0.8 nM 1370 nM 3200 nM
In vitro AZD8055 shows low activity (∼1,000-fold) against all PI3K isoforms (α, β, γ, δ) and other members of the PI3K-like kinase family (ATM and DNA-PK). AZD8055 inhibits the phosphorylation of mTORC1 (p70S6K and 4E-BP1) as well as phosphorylation of the mTORC2 (Akt) and downstream proteins. The rapamycin-resistant T37/46 phosphorylation sites on 4E-BP1 can be fully inhibited by AZD8055, resulting in significant inhibition of cap-dependent translation. AZD8055 potently inhibits proliferation in U87MG, A549 and H838 cells with IC50 of 53, 50 and 20 nM, respectively. AZD8055 also induces autophagy and increased LC3-II levels in H838 and A549 cells. [1] AZD8055 decreases AML blast cell proliferation and cell cycle progression, reduces the clonogenic growth of leukemic progenitors and induces caspase-dependent apoptosis in leukemic cells but not in normal immature CD34+ cells. [2] AZD8055 indicates inhibitory against the pediatric preclinical testing program (PPTP) cell lines with IC50 of 24.7 nM and induces significant differences in EFS distribution. [3]
Cell Data
Cell LinesAssay TypeConcentrationIncubation TimeFormulationActivity DescriptionPMID
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Kasumi-1NFTDXJJIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=M2Owd2lEPTB;MT60JI5ONF:xXIMzOTN|N{[3PS=>
Karpas-299M1XwUGdzd3e2aDDJcohq[mm2aX;uJGF{e2G7MWfJR|UxRTJzLkegcm0>MY[yNVM{PzZ5OR?=
Ramos-RA1NIP6UWNIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=M3H3dWlEPTB;OD6yJI5OMmHDNlE{Ozd4N{m=
H838M1Sxb2Z2dmO2aX;uJGF{e2G7NI\VS4gyKM7:TR?=NX7FWFRWPDhiaB?=MWXpcoR2[2W|IHPlcIwh[XW2b4DoZYd6NGjQXm0zODN4NEGxNy=>
H838M1\GUWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7M{LrXlAuOyEQvF2=M1rufFczKGh?M{n2Oolv\HWlZYOg[5Jwf3SqIHnubIljcXSrb36gZY5lKGG3dH;wbIFogQ>?NHvpXWgzODB{OEi1OC=>
A549 NYn1XoRwT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=NEXvRXUxNTNizszNNF7nbJQ4OiCqMoTBbY5lfWOnczDndo94fGhiaX7obYJqfGmxbjDhcoQh[XW2b4DoZYd6M1PRNVIxODJ6OEW0

... Click to View More Cell Line Experimental Data

In vivo AZD8055 inhibits the pS6 and pAkt in U87MG and A549 xenografts at 2.5/10 mg/kg, which leads to tumor growth inhibition. AZD8055 shows significant antitumor activity in many xenografts, including U87MG, BT474c, A549, Calu-3, LoVo, SW620, PC3 and MES-SA at a dose of 10/20 mg/kg. [1] AZD8055 induces ~40% reduction in tumour volume, accompanied by ablation of phosphorylation of Akt, S6K and SGK protein kinases. Administration of AZD8055 (5mg/kg, Bid) and SAHA (100 mg/kg/d) results in complete tumor growth inhibition in PTEN+/−LKB1+/hypo xenografts without side effects on mice by inhibition of mTORC1 and mTORC2 signaling. [4]
Features First drug to inhibit both types of mTOR protein.

Protocol(Only for Reference)

Kinase Assay: [1]

Cell-based determinations of mTOR inhibition A high-throughput screening cell-based assay is developed using MDA-MB-468 cells to detect mTORC1 and mTORC2 activity. Cells are exposed for 2 hours to increasing concentrations of AZD8055. At the end of the incubation period, cells are fixed, washed, and probed with antibodies against S473 pAkt or against S235/236 phosphorylated S6 (pS6). Levels of phosphorylation are assessed using an Acumen laser scanning cytometer.

Cell Assay: [1]

Cell lines U87MG, A549 and H838 cells
Concentrations 1 nM - 1 μM
Incubation Time 72 to 96 hours
Method Cells are exposed to AZD8055 for 72 to 96 hours and stained for cell nuclei (0.03 mg/mL Hoechst 33342) and acidic vesicles (1 μg/mL acridine orange). Images are captured at 450 and 536 nm on an ArrayScan II platform, and the percentage of acidic vesicles and the number of cells are quantified. For LC3 assessment, cells are exposed to e64d/pepstatin (10 μg/mL) for 30 to 90 min before incubation with AZD8055. Cells are lysed on ice and analyzed by immunoblotting.

Animal Study: [1]

Animal Models U87MG, BT474c, A549, Calu-3, LoVo, SW620, PC3 and MES-SA U87-MG and A549 are established in pathogen-free, female nude mice (nu/nu:Alpk).
Formulation Dissolved in captisol (CyDex, pH 3.0) and diluted to a final captisol concentration of 30% (w/v)
Dosages 2.5-20 mg/kg
Administration Oral gavage once or twice daily

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

SpeciesMouseRatRabbitGuinea pigHamsterDog
Weight (kg)0.020.151.80.40.0810
Body Surface Area (m2)0.0070.0250.150.050.020.5
Km factor36128520
Animal A (mg/kg) = Animal B (mg/kg) multiplied by  Animal B Km
Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×  mouse Km(3)  = 11.2 mg/kg
rat Km(6)
1

References

[1] Chresta CM, et al. Cancer Res, 2010, 70(1), 288-298.

[2] Willems L, et al. Leukemia, 2011, doi: 10.1038/leu.2011.339.

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Clinical Trial Information( data from http://clinicaltrials.gov, updated on 2016-07-23)

NCT Number Recruitment Conditions Sponsor
/Collaborators
Start Date Phases
NCT01316809 Completed Glioblastoma Multiforme|Anaplastic Astrocytoma|Anaplastic Oligodendroglioma|Malignant Glioma|Brainstem Glioma National Cancer Institute (NCI)|National Institutes of He  ...more National Cancer Institute (NCI)|National Institutes of Health Clinical Center (CC) March 2011 Phase 1
NCT00999882 Completed Cancer|Advanced Hepatocellular Carcinoma AstraZeneca October 2009 Phase 1
NCT00973076 Completed Cancer|Solid Tumors|Advanced Solid Malignancies AstraZeneca August 2009 Phase 1
NCT00731263 Completed Solid Tumors AstraZeneca July 2008 Phase 1
NCT01194193 Withdrawn Cancer|Advanced Solid Tumours|Lymphomas AstraZeneca null Phase 1

Chemical Information

Download AZD8055 SDF
Molecular Weight (MW) 465.54
Formula

C25H31N5O4

CAS No. 1009298-09-2
Storage 3 years -20℃powder
6 months-80℃in solvent
Synonyms N/A
Solubility (25°C) * In vitro DMSO 50 mg/mL warming (107.4 mM)
Ethanol 3 mg/mL (6.44 mM)
Water <1 mg/mL (<1 mM)
In vivo 4% DMSO+30% PEG 300+ddH2O 5mg/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.
Chemical Name (5-(2,4-bis((S)-3-methylmorpholino)pyrido[2,3-d]pyrimidin-7-yl)-2-methoxyphenyl)methanol

Customer Product Validation(7)


Click to enlarge
Rating
Source Cancer Res 2013 73(11), 3402-11. AZD8055 purchased from Selleck
Method Western blot
Cell Lines PC3-LN4 cells
Concentrations 1 uM
Incubation Time 24 h
Results GSK690693 was combined with two potent inhibitors of mTORC1/mTORC2 and thus cap-dependent translation, PP242 and AZD8055. AZD8055 in combination with GSK690693 resulted in reduced phosphorylation of 4E-BP1, and increased eIF2α phosphorylation compared to GSK690693 alone, suggesting inhibition of 5′-cap dependent translation.

Click to enlarge
Rating
Source Antioxid Redox Signal 2014 20(9), 1382-95. AZD8055 purchased from Selleck
Method Western blot
Cell Lines HUVEC
Concentrations 150 nM
Incubation Time 24 h
Results As indicated in Figure, inhibition of mTOR by AZD8055, an mTOR inhibitor, abolishes PA-suppressed PTEN nuclear localization, p53 acetylation, and GPX1 expression in ECs.

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Rating
Source Oncotarget 2014 5(15), 6015-27. AZD8055 purchased from Selleck
Method Western blot
Cell Lines HCT116 or MDA-MB-468 cells
Concentrations 500 nM
Incubation Time 12 h
Results AZD8055 effectively inhibited phosphorylation of 4E-BP1 at the four phosphorylation sites in the three tested cell lines (HCT116, MDA-MB-468 and MDA-MB-231). Inhibition of 4E-BP1 phosphorylation in these cells was associated with downregulation of Snail expression accompanied by an increase in E-cadherin expression upon AZD8055 exposure within 24 h.

Click to enlarge
Rating
Source Mol Cancer Ther 2014 13(1), 37-48. AZD8055 purchased from Selleck
Method MTS assay
Cell Lines CSCs<sup>+24/44/ESA</sup>
Concentrations 1 uM
Incubation Time 72 h
Results It found that BMS-777607 in combination with AZD8055 or PP242 decreased the percentages of CSCs+24/44/ESA-derived polyploid cells. It's suggest that inhibition of mTORC2 but not mTORC1 is primarily responsible for the observed synergistic effect on cell viability.

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Rating
Source Int J Cancer 2014 135(10), 2462-74. AZD8055 purchased from Selleck
Method Fluorescent assay
Cell Lines Serum-deprived CHO-EGFP-AKT cells
Concentrations 50 nM
Incubation Time 24 h
Results AZD8055 is a highly selective ATP-competitive inhibitor of mTOR, which inhibits phosphorylation of AKT at S473 by targeting mTORC2. The EGFP-AKT fusion protein largely remained in cytoplasm when cells were cultured in serum-free medium. By contrast, treatment of CHO-hIR cells with 50 nM AZD8055 for 24 hr resulted in translocation of AKT to the cellular membrane (indicated by white arrows), which was demonstrated as the fluorescent foci on the membrane, indicating activation of PI3K after mTOR inhibition.

Click to enlarge
Rating
Source Exp Neurol 2012 233, 799-806. AZD8055 purchased from Selleck
Method Western blotting
Cell Lines basilar artery
Concentrations
Incubation Time
Results Treatment with Rapamycin and AZD8055 significantly suppressed the expression of mTOR, P70S6K1, 4E-BP1, and PCNA.

Click to enlarge
Rating
Source Dr. Zhang of Tianjin Medical University. AZD8055 purchased from Selleck
Method Western blotting
Cell Lines A549 cells
Concentrations 0-10 µM
Incubation Time 3 h
Results AZD8055 treatment resulted in a reduction of Akt-473 phosphorylation in A549 cells.

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.

Tel: +1-832-582-8158 Ext:3

If you have any other enquiries, please leave a message.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
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