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

• ChemBase ID: 73225
• Molecular Formular: C22H27N5O4
• Molecular Mass: 425.48088
• Monoisotopic Mass: 425.20630437
• SMILES: n1c(nc2c(c1C)cc(c(=O)n2[C@H]1CC[C@@H](CC1)OCCO)c1ccc(nc1)OC)N
• Canonical SMILES: OCCO[C@@H]1CC[C@H](CC1)n1c(=O)c(cc2c1nc(N)nc2C)c1ccc(nc1)OC
• InChI: InChI=1S/C22H27N5O4/c1-13-17-11-18(14-3-8-19(30-2)24-12-14)21(29)27(20(17)26-22(23)25-13)15-4-6-16(7-5-15)31-10-9-28/h3,8,11-12,15-16,28H,4-7,9-10H2,1-2H3,(H2,23,25,26)/t15-,16-
• InChIKey: XDLYKKIQACFMJG-WKILWMFISA-N

NAMES AND DATABASE IDS

Names

• 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
• Synonyms: PF-04691502

Database IDs

• CAS Number: 1013101-36-4
• PubChem SID: 162038145
• PubChem CID: 25033539

CALCULATED PROPERTIES

• Acid pKa: 15.106726
• H Acceptors: 8
• H Donor: 2
• LogD (pH = 5.5): 1.3600361
• LogD (pH = 7.4): 1.3660862
• Log P: 1.366164
• Molar Refractivity: 117.4099 cm^3
• Polarizability: 43.91023 Å^3
• Polar Surface Area: 123.69 Å^2
• Rotatable Bonds: 6
• Lipinski's Rule of Five: true

PROPERTIES

Safety Information

• Storage Condition: -20°C

Pharmacology Properties

• Target: mTOR / PI3K

Product Information

• Salt Data: Free Base

DETAILS

Selleck Chemicals - S2743

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