| Size | Price | Stock | Qty |
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| 10mg |
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| Other Sizes |
| Targets |
CENP-E 16.1 nM (IC50)
PF-2771 specifically targets the centromere protein E (CENP-E), a mitotic kinesin (kinesin-7) that plays an essential role in the spindle assembly checkpoint and chromosome alignment during mitosis. The compound inhibits CENP-E motor activity with an IC50 of 16.1 nM. PF-2771 shows no inhibitory effect on ATPase activities of highly related kinesins (0% inhibition of Eg5/KSP, chromokinesin, and MCAK at both 1 or 10 microM), demonstrating excellent selectivity within the kinesin superfamily. |
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| ln Vitro |
With an IC50 of 16.1 nM, PF-2771 is a strong and specific inhibitor of CENP-E that inhibits CENP-E motor activity. The ATPase activities of closely related kinesins are not inhibited by PF-2771 (0% inhibition of MCAK, chromokinesin, and Eg5/KSP at both 1 and 10 μM PF-2771). 74 protein kinases are inactively inhibited by PF-2771 (all <23% inhibition at 1 μM, <40% at 10 μM PF-2771). With EC50s of less than 0.1 μM, PF-2771 is cytotoxic to the basal-like breast cancer tumor cell survival; however, it has no effect on normal or premalignant cell lines (EC50 > 5 μM). In MDA-MB-468 cells, PF-2771 (100 nM) causes a chromosomal congression defect[1].
In vitro, PF-2771 (IC50 = 16.1 nM) potently and selectively inhibits CENP-E motor activity. It exhibits inactive activity against 74 protein kinases (all <23% inhibition at 1 microM, <40% at 10 microM). PF-2771 is cytotoxic to basal-like breast cancer tumor cell survival with EC50s of <0.1 microM, while having no effect on normal or premalignant cell lines (EC50 >5 microM). In MDA-MB-468 cells, PF-2771 (100 nM) causes chromosomal congression defects during mitosis, linking CENP-E-dependent molecular events such as histone H3 Ser-10 phosphorylation (phospho-HH3-Ser10) to functional mitotic defects. |
| ln Vivo |
In SCID mice with AA1077 mammary tumors, PF-2771 (100 mg/kg, intraperitoneally every day) potently suppresses CENP-E motor activity and promotes tumor regression[1].
In vivo, PF-2771 demonstrates potent antitumor efficacy. In SCID mice bearing AA1077 mammary tumors, PF-2771 (100 mg/kg, intraperitoneal injection daily) potently suppresses CENP-E motor activity and promotes tumor regression. Complete tumor regression is observed in a patient-derived basal-a breast cancer xenograft model treated with PF-2771. Tumor regression is also observed with PF-2771 in a taxane-resistant basal-a model. These findings suggest CENP-E may be an effective therapeutic target for triple-negative/basal-a breast cancer. |
| Enzyme Assay |
To assess CENP-E motor inhibition in a cell-free system, a biochemical ATPase assay is used. Purified recombinant CENP-E motor domain is incubated with varying concentrations of PF-2771 (0, 2.5, 5, 10, 20, 30, 45 nM) in reaction buffer containing ATP at concentrations ranging from 0.977 to 1000 microM. The enzymatic activity is measured by quantifying the amount of inorganic phosphate released using a colorimetric or malachite green-based detection method. The IC50 value (16.1 nM) is calculated from the dose-response inhibition curve.
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| Cell Assay |
For cellular assays, basal-like breast cancer cell lines such as MDA-MB-468 are seeded in 96-well plates and treated with serial dilutions of PF-2771 (0.01 to 10 microM) for 48-72 hours. Cell viability is measured using the CellTiter-Glo luminescent assay to determine EC50 values. For mechanistic studies, cells are treated with PF-2771 (e.g., 100 nM) and fixed. Immunofluorescence staining for histone H3 phosphorylated at Ser-10 (phospho-HH3-Ser10) is performed to monitor chromosome congression defects. Chromosomal alignment is assessed by confocal microscopy.
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| Animal Protocol |
For in vivo efficacy studies, immunocompromised mice (SCID mice) bearing AA1077 mammary tumor xenografts are used. PF-2771 is formulated in a suitable vehicle such as 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline at a concentration of 10 mg/mL. The compound is administered via intraperitoneal injection daily at a dose of 100 mg/kg. Tumor volume is measured with calipers every 2-3 days. Endpoints include tumor growth inhibition (TGI) and tumor regression assessment. Tumors are harvested for Western blot analysis of phospho-HH3-Ser10 to confirm target engagement.
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| ADME/Pharmacokinetics |
Pharmacokinetic analysis in a basal-a xenograft tumor model shows that PF-2771 exposure is well correlated with increased phospho-HH3-Ser10 levels and tumor growth regression, indicating favorable pharmacodynamic properties. The compound is soluble in DMSO (≥32 mg/mL, 57.75 mM). For in vivo administration, it can be formulated in 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline (≥2.5 mg/mL, 4.51 mM) to achieve systemic exposure sufficient for efficacy. Predicted logP: 3.8, tPSA: 106, density: 1.22+/-0.1 g/cm3.
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| Toxicity/Toxicokinetics |
Specific toxicology data for PF-2771 is not detailed in the literature. However, its highly selective mechanism targeting CENP-E during mitosis suggests on-target toxicity may be primarily associated with rapidly dividing cells, as observed in preclinical efficacy studies where the compound was tolerated at doses up to 100 mg/kg daily in SCID mice. No significant weight loss or acute toxicity was reported at efficacious doses, indicating a manageable safety profile for a research compound. Its selectivity over 74 protein kinases also suggests minimal off-target kinase-related toxicity.
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| References | |
| Additional Infomation |
PF-2771 was developed by Pfizer Worldwide Research and Development as a first-in-class potent and selective chemical probe for CENP-E. The compound's mechanism of action involves inhibiting CENP-E motor ATPase activity, which disrupts proper chromosome congression during mitosis, leading to mitotic arrest and cell death specifically in cancer cells with high levels of chromosomal instability. This selectivity provides a unique therapeutic window for treating basal-like/triple-negative breast cancer. PF-2771 is a preclinical research compound; no clinical trials or FDA approval has been reported for this agent.
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| Molecular Formula |
C29H36CLN5O4
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|---|---|
| Molecular Weight |
554.08
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| Exact Mass |
553.245
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| CAS # |
2070009-55-9
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| PubChem CID |
118704759
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| Appearance |
White to off-white solid powder
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| Density |
1.22±0.1 g/cm3(Predicted)
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| LogP |
3.8
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
12
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| Heavy Atom Count |
39
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| Complexity |
820
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CC(C)OC1=C(C=C(C=C1)C(=O)N[C@@H](CC2=CC=C(C=C2)C3=CN(C(=N3)C(=O)C)C)CNC(=O)CN(C)C)Cl
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| InChi Key |
VHBJNUOZEQTSNN-QHCPKHFHSA-N
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| InChi Code |
InChI=1S/C29H36ClN5O4/c1-18(2)39-26-12-11-22(14-24(26)30)29(38)32-23(15-31-27(37)17-34(4)5)13-20-7-9-21(10-8-20)25-16-35(6)28(33-25)19(3)36/h7-12,14,16,18,23H,13,15,17H2,1-6H3,(H,31,37)(H,32,38)/t23-/m0/s1
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| Chemical Name |
N-[(2S)-1-[4-(2-acetyl-1-methylimidazol-4-yl)phenyl]-3-[[2-(dimethylamino)acetyl]amino]propan-2-yl]-3-chloro-4-propan-2-yloxybenzamide
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO: ≥ 32 mg/mL (57.75 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.51 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (4.51 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (4.51 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8048 mL | 9.0240 mL | 18.0479 mL | |
| 5 mM | 0.3610 mL | 1.8048 mL | 3.6096 mL | |
| 10 mM | 0.1805 mL | 0.9024 mL | 1.8048 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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