PF-04691502

• Catalog No.: S2743
• CAS Number: 1013101-36-4
• M. F.: C22H27N5O4
• M. W.: 425.48088
• Storage: -20°C

SYNONYMS

• IUPAC name: 2-amino-6-(6-methoxypyridin-3-yl)-4-methyl-8-[(1r,4r)-4-(2-hydroxyethoxy)cyclohexyl]-7H,8H-pyrido[2,3-d]pyrimidin-7-one
• IUPAC Traditional name: 2-amino-6-(6-methoxypyridin-3-yl)-4-methyl-8-[(1r,4r)-4-(2-hydroxyethoxy)cyclohexyl]pyrido[2,3-d]pyrimidin-7-one

DATABASE IDS

• CAS Number: 1013101-36-4

PROPERTIES

• Target: mTOR / PI3K
• Salt Data: Free Base
• Storage Condition: -20°C

DETAILS

Description (English)

Biological Activity

• Description: PF-04691502 is an ATP-competitive PI3K/mTOR dual inhibitor with IC50 of 32 nM.
• Targets: mTORC1
• IC50: 32 nM ^[1]
• In Vitro: PF-04691502 potently inhibits recombinant class I PI3K and mTOR in biochemical assays and suppresses transformation of avian fibroblasts mediated by wild-type PI3K γ, δ, or mutant PI3Kα. In PIK3CA-mutant and PTEN-deleted cancer cell lines, PF-04691502 reduces phosphorylation of AKT T308 and AKT S473 (IC(50) of 7.5-47 nM and 3.8-20 nM, respectively) and inhibits cell proliferation (IC(50) of 179-313 nM). PF-04691502 inhibits mTORC1 activity in cells as measured by PI3K-independent nutrient stimulated assay, with an IC(50) of 32 nM and inhibits the activation of PI3K and mTOR downstream effectors including AKT, FKHRL1, PRAS40, p70S6K, 4EBP1, and S6RP. Short-term exposure to PF-04691502 predominantly inhibits PI3K, whereas mTOR inhibition persists for 24 to 48 hours. PF-04691502 induces cell cycle G(1) arrest, concomitant with upregulation of p27 Kip1 and reduction of Rb. ^[1]
• In Vivo: Antitumor activity of PF-04691502 is observed in U87 (PTEN null), SKOV3 (PIK3CA mutation), and gefitinib- and erlotinib-resistant non-small cell lung carcinoma xenografts. ^[1] PF-04691502 inhibits tumor growth at 7 days by 72%. FDG-PET imaging revealed that PF-04691502 reduces glucose metabolism dramatically. Tissue biomarkers of PI3K/mTOR pathway activity, p-AKT (S473), and p-RPS6 (S240/244), are also dramatically inhibited following PF-04691502 treatment. ^[2]
• Clinical Trials: PF-04691502 is currently in Phase II clinical trials in patients with recurrent endometrial cancer and Breast Cancer. A Phase II clinical trials in patients with solid tumors has been completed.

Combination Therapy

• Description: Combination with PF-04691502, PD-0325901 induces striking tumor regression, apoptosis associated with upregulation of Bim and downregulation of Mcl-1, and greatly improves duration of survival. ^[2] PD-0325901 at 1.5 mg/kg SID and in combination with PF-04691502 (7.5 mg/kg; SID) significantly inhibits growth of H460 (carry Kras and PIK3CA mutations) orthotopic lung tumors. ^[3]

Protocol

• Protocol: Kinase Assay ^[1]
• Kinase Assay: The fluorescence polarization assay for ATP competitive inhibition is done as follows: mPI3Kα dilution solution (90 nM) is prepared in fresh assay buffer (50 mM Hepes pH 7.4, 150 mM NaCl, 5 mM DTT, 0.05% CHAPS) and kept on ice. The enzyme reaction contains 0.5 nM mouse PI3Kα (p110α/p85α complex purified from insect cells), 30 μM PIP2, PF-04691502 (0, 1, 4, and 8 nM), 5 mM MgCl₂, and 2-fold serial dilutions of ATP (0–800 μM). Final dimethyl sulfoxide is 2.5%. The reaction is initiated by the addition of ATP and terminated after 30 minutes with 10 mM EDTA. In a detection plate, 15 uL of detector/probe mixture containing 480 nM GST-Grp1PH domain and 12 nM TAMRA tagged fluorescent PIP3 in assay buffer is mixed with 15 uL of kinase reaction mixture. The plate is shaken for 3 minutes, and incubated for 35 to 40 minutes before reading on an LJL Analyst HT.
• Protocol: Cell Assay ^[1]
• Cell Lines: BT20, U87MG, and SKOV3 cells
• Concentrations: 0-3 mM
• Incubation Time: 3 days
• Methods: BT20, U87MG, and SKOV3 cells are plated at 3,000 cell/well in 96-well culture plates in growth medium with 10% FBS. Cells are incubated overnight and treated with DMSO (0.1% final) or serial diluted compound for 3 days. Resazurin is added to 0.1 mg/mL. Plates are incubated at 37 °C in 5% CO₂ for 3 hours. Fluorescence signals are read as emission at 590 nm after excitation at 530 nm. IC50 values are calculated by plotting fluorescence intensity to drug concentration in nonlinear curves.
• Protocol: Animal Study ^[2]
• Animal Models: LSL-Kras^G12D heterozygous mice (B6.129-Kras^tm4Tyj) and Pten^del mice (c;129S4-Pten^tm1Hwu/J), Orthotopic transplant of ovarian tumors
• Formulation: 0.5% methylcellulose
• Doses: daily at either 7.5 or 10 mg/kg
• Administration: Administered via oral gavage

References

• References: [1] Yuan J, Mol Cancer Ther, 2011, 10(11), 2189-2199
• References: [2] Kinross KM, Mol Cancer Ther, 2011, 10(8), 1440-1449
• References: [3] Simmons BH, Cancer Chemother Pharmacol, 2012, 70(2), 213-220

REFERENCES

• Yuan J, Mol Cancer Ther, 2011, 10(11), 2189-2199
• Kinross KM, Mol Cancer Ther, 2011, 10(8), 1440-1449
• Simmons BH, Cancer Chemother Pharmacol, 2012, 70(2), 213-220