Palbociclib (PD-0332991) HCl Licensed by Pfizer

Catalog No.S1116

Palbociclib (PD-0332991) HCl is a highly selective inhibitor of CDK4/6 with IC50 of 11 nM/16 nM in cell-free assays, respectively. It shows no activity against CDK1/2/5, EGFR, FGFR, PDGFR, InsR, etc. Phase 3.

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Palbociclib (PD-0332991) HCl Chemical Structure

Palbociclib (PD-0332991) HCl Chemical Structure
Molecular Weight: 483.99

Validation & Quality Control

Customer Product Validation(6)

Quality Control & MSDS

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Product Information

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  • Research Area
  • Inhibition Profile

Product Description

Biological Activity

Description Palbociclib (PD-0332991) HCl is a highly selective inhibitor of CDK4/6 with IC50 of 11 nM/16 nM in cell-free assays, respectively. It shows no activity against CDK1/2/5, EGFR, FGFR, PDGFR, InsR, etc. Phase 3.
Targets CDK4/CyclinD3 [1]
(Cell-free assay)
CDK4/CyclinD1 [1]
(Cell-free assay)
CDK6/CyclinD2 [1]
(Cell-free assay)
CDK2/CyclinE2 [1]
(Cell-free assay)

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IC50 9 nM 11 nM 15 nM >10 μM
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 Ser780. 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 0.04-0.17 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]
Cell Data
Cell LinesAssay TypeConcentrationIncubation TimeFormulationActivity DescriptionPMID
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... Click to View More Cell Line Experimental Data

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]
Features Non-cytotoxic, and halts cancer cell growth & could be used in glioblastoma that has relapsed after temozolomide treatment (a chemotherapeutic used to treat many cancers).

Protocol(Only for Reference)

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.

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
Method Cells are seeded at 2 × 104 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.

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)
Dosages 0-150 mg/kg
Administration Given by gavage

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] Fry DW, et al. Mol Cancer Ther, 2004, 3(11), 1427-1438.

[2] Menu E, et al. Cancer Res, 2008, 68(14), 5519-5523.

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Clinical Trial Information( data from http://clinicaltrials.gov, updated on 2016-05-07)

NCT Number Recruitment Conditions Sponsor
/Collaborators
Start Date Phases
NCT02730429 Not yet recruiting Endometrial Cancer Nordic Society for Gynaecologic Oncology|ENGOT|GCIG August 2016 Phase 2
NCT02738866 Not yet recruiting Metastatic Breast Cancer Sidney Kimmel Comprehensive Cancer Center|Pfizer July 2016 Phase 2
NCT02764541 Not yet recruiting Breast Cancer Dana-Farber Cancer Institute|Pfizer May 2016 Phase 2
NCT02684032 Not yet recruiting Breast Cancer Pfizer May 2016 Phase 1
NCT02600923 Recruiting Advanced Breast Cancer (Female) Pfizer April 2016 Phase 3

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Chemical Information

Download Palbociclib (PD-0332991) HCl SDF
Molecular Weight (MW) 483.99
Formula

C24H29N7O2.HCl

CAS No. 827022-32-2
Storage 3 years -20℃powder
6 months-80℃in solvent
Synonyms N/A
Solubility (25°C) * In vitro DMSO 3 mg/mL warmed (6.19 mM)
Water 30 mg/mL (61.98 mM)
Ethanol <1 mg/mL (<1 mM)
In vivo Saline 20 mg/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 6-acetyl-8-cyclopentyl-5-methyl-2-(5-(piperazin-1-yl)pyridin-2-ylamino)pyrido[2,3-d]pyrimidin-7(8H)-one hydrochloride

Customer Product Validation (6)


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Rating
Source Clin Cancer Res 2011 17(13), 4513-22. Palbociclib (PD-0332991) HCl purchased from Selleck
Method Western Blot/Growth assay
Cell Lines OE33/OE19/Flo1A cells
Concentrations 125 nM
Incubation Time 1/3/21 day
Results PD-0332991 reduces expression of cyclinA in all 3 cell lines. PD-0332991 has no effect on the expression level of either total pRB or RB2 (p130) in EAC cells. PD-0332991 caused a significant reduction in colony formation in soft agar in all 3 cell lines tested, indicative of a reduction in anchorage independence.

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Rating
Source Pharmacogenomics J 2013 13(1), 94-104. Palbociclib (PD-0332991) HCl purchased from Selleck
Method Immunofluorescence
Cell Lines Patient tumor cell
Concentrations 3 uM
Incubation Time
Results Tumor organoids treated with VPA/ PD0332991 or SAHA/PD0332991 exhibited reduced structural integrity and increased cellular scatter with increasing concentrations of the drugs.

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Rating
Source Clin Transl Oncol 2012 14(3), 197-206 . Palbociclib (PD-0332991) HCl purchased from Selleck
Method Immunofluorescence
Cell Lines Murine MSCs cells
Concentrations 2 uM
Incubation Time 24-48 h
Results We show that when cells are treated with PD332991 for 24–48 h there is a marked decrease in nuclear expression of cdk4 in PAX7- FKHR-expressing mMSCs. Further analysis by immunofluorescence showed that there is a minor upregulation in expression of MyoD1 in cells treated with cdk4 inhibitor for 48 h.

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Rating
Source Dr. Gao Zhang of University of Pennsylvania. Palbociclib (PD-0332991) HCl purchased from Selleck
Method Western blot
Cell Lines WM3734 melanoma cells
Concentrations 0.25-4 μM
Incubation Time 36 h
Results

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Rating
Source Dr. Sabine Paternot and Dr Pierre Roger of IRIBHM, ULB, Brussels. Palbociclib (PD-0332991) HCl purchased from Selleck
Method Western blot
Cell Lines T98G glioma cells
Concentrations 1 μM
Incubation Time
Results

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Rating
Source Dr. Kun-Wei Liu from University of Pittsburgh. Palbociclib (PD-0332991) HCl purchased from Selleck
Method IB analysis, Quantification of soft agar assays
Cell Lines LN319 cells
Concentrations
Incubation Time
Results

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

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
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