5-[3-(benzyloxy)phenyl]-7-[(1s,3s)-3-(azetidin-1-ylmethyl)cyclobutyl]-7H-pyrrolo[2,3-d]pyrimidin-4-amine

• ChemBase ID: 73155
• Molecular Formular: C27H29N5O
• Molecular Mass: 439.55206
• Monoisotopic Mass: 439.23721057
• SMILES: c1cc(cc(c1)OCc1ccccc1)c1cn(c2c1c(ncn2)N)[C@@H]1C[C@@H](C1)CN1CCC1
• Canonical SMILES: Nc1ncnc2c1c(cn2[C@@H]1C[C@@H](C1)CN1CCC1)c1cccc(c1)OCc1ccccc1
• InChI: InChI=1S/C27H29N5O/c28-26-25-24(21-8-4-9-23(14-21)33-17-19-6-2-1-3-7-19)16-32(27(25)30-18-29-26)22-12-20(13-22)15-31-10-5-11-31/h1-4,6-9,14,16,18,20,22H,5,10-13,15,17H2,(H2,28,29,30)/t20-,22+
• InChIKey: AECDBHGVIIRMOI-GRGXKFILSA-N

NAMES AND DATABASE IDS

Names

• IUPAC name: 5-[3-(benzyloxy)phenyl]-7-[(1s,3s)-3-(azetidin-1-ylmethyl)cyclobutyl]-7H-pyrrolo[2,3-d]pyrimidin-4-amine
• IUPAC Traditional name: 5-[3-(benzyloxy)phenyl]-7-[(1s,3s)-3-(azetidin-1-ylmethyl)cyclobutyl]pyrrolo[2,3-d]pyrimidin-4-amine
• Synonyms: AEW541; NVP-AEW541

Database IDs

• CAS Number: 475489-16-8
• PubChem SID: 162038075
• PubChem CID: 11476171

CALCULATED PROPERTIES

• H Acceptors: 5
• H Donor: 1
• LogD (pH = 5.5): -0.7197423
• LogD (pH = 7.4): 2.015645
• Log P: 4.1548033
• Molar Refractivity: 132.7197 cm^3
• Polarizability: 52.0621 Å^3
• Polar Surface Area: 69.2 Å^2
• Rotatable Bonds: 7
• Lipinski's Rule of Five: true

PROPERTIES

Safety Information

• Storage Condition: -20°C

Pharmacology Properties

• Target: IGF-1R

Product Information

• Salt Data: Free Base

DETAILS

Selleck Chemicals - S1034

Biological Activity

• Description: NVP-AEW541 is a potent inhibitor of IGF-IR with IC50 of 86 nM.
• Targets: IGF-IR
• IC50: 86 nM ^[1]
• In Vitro: NVP-AEW541 also inhibits InsR, Tek, Flt1 and Flt3 with IC50 of 140 nM, 530 nM, 600 nM and 420 nM in purified kinases/recombinant kinase domains assay. NVP-AEW541 is more selective and shows 27-fold more potent than InsR at the cellular level. NVP-AEW541 suppresses the IGF-I-mediated survival, soft agar and proliferation of MCF-7 cells with IC50 of 0.162 μM, 0.105 μM and 1.64 μM, respectively. NVP-AEW541 also reduces the level of phospho-IGF-IR and phospho-PKB in NWT-21 cells. ^[1] NVP-AEW541 shows growth inhibitory effect on TC-71 musculoskeletal sarcoma cells in low-serum medium as well as in 10% FBS–containing medium. NVP-AEW541 inhibits cell cycle progression and induces specific G1 arrest in sarcoma cell lines (TC-71, SK-N-MC, SaoS-2, RD/18 and RH4). ^[2] NVP-AEW541 could inhibit the growth of human neuroblastoma cells with IC50 of 0.4-6.8 μM. An increase in the hypodiploid fraction and the depletion of the S and G2-M compartments could be detected in these cell lines. NVP-AEW541-driven inhibition of IGF-IR causes a reduction of phosphorylation of Akt, but not of Erk1 and Erk2 in neuroblastoma cells. ^[3] NVP-AEW541 inhibits glioma cell growth and disrupts the autocrine loop initiated by HIF1α stabilization. ^[4] A recent study shows that NVP-AEW541 suppresses the proliferation and viability of PC3, DU145, and 22Rv1 prostate cancer cells, without necessarity of associated cell death. NVP-AEW541 decreases phospho-Akt levels in 22Rv1 and DU415 cells but not PC3 cells, without affecting total Akt levels, which shows that PTEN status could determine the effectiveness of NVP-AEW541 with essential Akt. NVP-AEW541-induced radiosensization is dependent on Akt activation status. NVP-AEW541 could increase the H2AX phosphorylation (a measure of DSBs) in PC3, DU145, and 22Rv1 cells. ^[5]
• In Vivo: NVP-AEW541 (50 mg/kg, p.o.) results in abrogation of basal and IGF-I-induced receptor, and PKB and MAPK phosphorylation, with T/C value of 14% in the NWT-21 tumor xenograft. ^[1] NVP-AEW541 (50 mg/kg) causes tumor shrinkage in both HTLA-230 and SK-N-BE2c xenografts, without signs of systemic toxicity. NVP-AEW541 could inhibit tumor invasion both in Matrigel-coated chambers and in HTLA-230 xenografts. ^[3]

Protocol

• Protocol: Kinase Assay ^[1]
• In vitro kinase assays: NVP-AEW541 is dissolved in DMSO (10 mM) and stored at -20 °C. Dilutions are freshly made in DMSO/water 1:1. The final concentration of DMSO in the enzyme assays is <0.5 %.="" the="" protein="" kinase="" assays="" are="" carried="" out="" in="" 96-well="" plates="" at="" rt="" and="" terminated="" by="" the="" addition="" of="" 20="" μl="" of="" 125="" mm="" edta.="" subsequently,="" 30="" μl="" (c-abl,="" c-src,="" igf-1r)="" of="" the="" reaction="" mixture="" are="" transferred="" onto="" immobilon-pvdf="" presoaked="" for="" 5="" min="" with="" methanol,="" rinsed="" with="" water,="" then="" soaked="" for="" 5="" min="" with="" 0.5="" %="">₃PO₄ and mounted on vacuum manifold. After spotting all samples, vacuum is connected and each well rinsed with 200 μL 0.5 % H3PO4. Membranes are removed and washed 4× on a shaker with 1.0 % H₃PO₄, once with ethanol. After drying, mounting in Packard TopCount 96-well frame, and adding of 10 μL/well of Microscint, membranes are counted. IC50 values are calculated by linear regression analysis of the percentage inhibition of NVP-AEW541 in duplicate, at four concentrations (usually 0.01, 0.1, 1, and 10 μM). One unit of protein kinase activity is defined as 1 nmol of ^33P transferred from [γ^33P]ATP to the substrate protein per minute per mg of protein at 37 °C.
• Protocol: Cell Assay ^[1]
• Cell Lines: MCF-7 cells
• Concentrations: ~ 10 μM
• Incubation Time: 72 hours
• Methods: Between 3 × 10^3 and 6 × 10^3 cells/well are seeded in 96-well plates with a total media volume of 100 μL/well. Increasing concentrations of NVP-AEW541 is added 24 hours thereafter in quadruplicate. 72 hours later, cells are fixed by addition of 25 μL/well Glutaraldehyde (20%) and incubation for 10 min at RT. Cells are then washed 2× with 200 μL/well H₂O and 100 μL Methylene Blue (0.05%) is added. After incubation for 10 min at RT, cells are washed 3× with 200 μL/well H₂O. 200 μL/well HCl (3%) is added, and following incubation for 30 min at RT on a plate shaker, absorbance is measured at 650 nm.
• Protocol: Animal Study ^[1]
• Animal Models: Female Harlan athymic nude mice weighing 18-25 g with NWT-21 cells
• Formulation: Dissolved in 25 mM L(+)-tartaric acid
• Doses: 20, 30, or 50 mg/kg
• Administration: Administered via p.o. twice daily

References

• References: [1] García-Echeverría C, et al. Cancer Cell. 2004, 5(3), 231-239.
• References: [2] Scotlandi K, et al. Cancer Res, 2005, 65(9), 3868-3876.
• References: [3] Tanno B, et al. Clin Cancer Res. 2006, 12(22), 6772-6780.
• References: [4] Gariboldi MB, et al. Biochem Pharmacol, 2010, 80(4), 455-462.
• References: [5] Isebaert SF, et al. Int J Radiat Oncol Biol Phys, 2011, 81(1), 239-247.

REFERENCES

• García-Echeverría C, et al. Cancer Cell. 2004, 5(3), 231-239.
• Scotlandi K, et al. Cancer Res, 2005, 65(9), 3868-3876.
• Tanno B, et al. Clin Cancer Res. 2006, 12(22), 6772-6780.
• Gariboldi MB, et al. Biochem Pharmacol, 2010, 80(4), 455-462.
• Isebaert SF, et al. Int J Radiat Oncol Biol Phys, 2011, 81(1), 239-247.