PD 0332991

• Catalog No.: S1116
• CAS Number: 827022-32-2
• M. F.: C24H30ClN7O2
• M. W.: 483.9937
• Storage: -20°C

SYNONYMS

• IUPAC name: 6-acetyl-8-cyclopentyl-5-methyl-2-{[5-(piperazin-1-yl)pyridin-2-yl]amino}-7H,8H-pyrido[2,3-d]pyrimidin-7-one hydrochloride
• IUPAC Traditional name: 6-acetyl-8-cyclopentyl-5-methyl-2-{[5-(piperazin-1-yl)pyridin-2-yl]amino}pyrido[2,3-d]pyrimidin-7-one hydrochloride

DATABASE IDS

• CAS Number: 827022-32-2

PROPERTIES

• Target: CDK
• Salt Data: hydrochloride
• Solubility: DMSO
• Storage Condition: -20°C

DETAILS

Description (English)

Biological Activity

• Description: PD 0332991 is a highly selective inhibitor of CDK4/cyclin D1 and CDK6/cyclin D2 with IC50 of 11 nM and 16 nM, respectively.
• Targets: CDK4/cyclin D1; CDK4/cyclin D2
• IC50: 11 nM; 16 nM ^[1]
• In Vitro: PD 0332991 has little effect on other protein kinases including EGFR, FGFR, PGFR, IR. PD 0332991 is a non-ATP competitive inhibitor of Cdk4. PD 0332991 inhibits MDA-MB-435 breast carcinoma cells with IC50 of 66 nM, which is due to reduced Rb phosphorylation at Ser^780. PD 0332991 inhibits thymidine incorporation into the DNA of Rb-positive human breast, colon, and lung carcinomas as well as human leukemias, with IC50 values ranging from 0.04 to 0.17 μM. PD 0332991 shows no activity in Rb-negative cells. PD 0332991 causes an accumulation of cells in G1 in MDA-MB-453 breast and Colo-205 carcinoma cells. ^[1] PD 0332991 also shows activity in 5T33MM myeloma cells (immunocompetent model) and sensitizes the cells to killing by bortezomib. ^[2] PD 0332991 inhibits luminal ER-positive as well as HER2-amplified breast cancer cell lines including MDA-MB-175, ZR-75-30, CAMA-1, MDA-MB-134, HCC-202 and UACC-893. PD 0332991 enhances the activity of tamoxifen and trastuzumab in these cell lines. PD 0332991 enhances the sensitivity of tamoxifen in the MCF7 tamoxifen-resistant cells. ^[3] A recent study shows that PD 0332991 could suppress malignant rhabdoid tumor (MRT) cell lines including MP-MRT-AN, KP-MRT-RY, G401, KP-MRT-NS and the sensitivity of the MRT cell lines to PD 0332991 is inversely correlated with expression of p16. ^[4]
• In Vivo: PD 0332991 indicates complete tumor stasis in a MDA-MB-435 xenograft at 150 mg/kg. PD 0332991 also shows broad-spectrum antitumor activity in multiple human tumor xenografts by eliminating phospho-Rb and the proliferative marker Ki-67 from tumor tissue and down-regulation of genes under the transcriptional control of E2F. ^[1]
• Clinical Trials: PD 0332991 is currently under Phase II clinical trial for advanced or metastatic liposarcoma.
• Features: PD 0332991 itself does not kill cancer cells - just halts their growth and could be used in which glioblastoma has come back after treatment with temozolomide, a chemotherapy used in many cancer patients.

Protocol

• Protocol: Kinase Assay ^[1]
• Cdk Assays: A stock solution of PD0332991 is prepared in DMSO. CDK assays are performed in 96-well filter plates. All CDK-cyclin kinase complexes are expressed in insect cells through baculovirus infection and purified. The substrate for the assays is a fragment (amino acids 792–928) of pRb fused to GST (GST·RB-Cterm). The total volume in each well is 0.1 mL containing a final concentration of 20 mM Tris-HCl, pH 7.4, 50 mM NaCl, 1 mM dithiothreitol, 10 mM MgCl₂, 25 μM ATP (for CDK4-cyclin D1, CDK6-cyclin D2, and CDK6-cyclin D3) or 12 μM ATP (for CDK2-cyclin E, CDK2-cyclin A, and CDC2-cyclin B) containing 0.25 μCi of [γ-^32P]ATP, 20 ng of enzyme, 1 μg of GST·RB-Cterm, and PD 0332991 (0.001-0.1μM). All components except the [γ-^32P]ATP are added to the wells, and the plate is placed on a plate mixer for 2 min. The reaction is started by adding the [γ-^32P]ATP and the plate is incubated at 25 °C for 15 min. The reaction is terminated by addition of 0.1 mL of 20% trichloroacetic acid and the plate is kept at 4 ?°C for at least 1 hour to allow the substrate to precipitate. The wells are then washed 5 times with 0.2 mL of 10% trichloroacetic acid and radioactive incorporation is determined with a β plate counter.
• Protocol: Cell Assay ^[1]
• Cell Lines: Tumor cell lines including MDA-MB-435, ZR-75-1, T-47D, MCF-7, H1299, Colo-205, MDA-MB-468, H2009, CRRF-CEM and K562
• Concentrations: 0.01-1 μM
• Incubation Time: 24 hours
• Methods: Cells are seeded at 2 × 10^4 per well in a 96-well plate and incubated overnight. PD 0332991 (0.01-1 μM) is added to the wells and incubated at 37 °C for another 24 hours. [^14C]Thymidine (0.1 μCi) is added to each well and incorporation of the radiolabel is allowed to proceed for 72 hours. Incorporated radioactivity is determined with a β plate counter.
• Protocol: Animal Study ^[1]
• Animal Models: Advanced stage human tumor xenografts including Colo-205, MDA-MB-435 breast, SF-295 glioblastoma, ZR-75-1 breast, PC-3 prostate, H125 lung, SW-620 colon, H23 lung and MDA-MB-468 breast (Rb negative) are established in severe combined immunodeficient mice.
• Formulation: Dissolved in sodium lactate buffer (50 mM, pH 4.0)
• Doses: 0-150 mg/kg
• Administration: Given by gavage

References

• References: [1] Fry DW, et al. Mol Cancer Ther, 2004, 3(11), 1427-1438.
• References: [2] Menu E, et al. Cancer Res, 2008, 68(14), 5519-5523.
• References: [3] Finn RS, et al. Breast Cancer Res, 2009, 11(5), R77.
• References: [4] Katsumi Y, et al. Biochem Biophys Res Commun, 2011, 413(1), 62-68.

REFERENCES

• Fry DW, et al. Mol Cancer Ther, 2004, 3(11), 1427-1438.
• Menu E, et al. Cancer Res, 2008, 68(14), 5519-5523.
• Finn RS, et al. Breast Cancer Res, 2009, 11(5), R77.
• Katsumi Y, et al. Biochem Biophys Res Commun, 2011, 413(1), 62-68.