1. Neuronal Signaling
    Stem Cell/Wnt
  2. γ-secretase

PF-3084014 (Synonyms: Nirogacestat; PF-03084014)

Cat. No.: HY-15185 Purity: 99.95%
Data Sheet SDS Handling Instructions

PF-3084014 is a reversible, noncompetitive, and selective γ-secretase inhibitor with IC50 of 6.2 nM.

For research use only. We do not sell to patients.
PF-3084014 Chemical Structure

PF-3084014 Chemical Structure

CAS No. : 1290543-63-3

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Free Sample (0.5-1 mg)   Apply now  
10 mM * 1 mL in DMSO $198 In-stock
5 mg $180 In-stock
10 mg $310 In-stock
50 mg $900 In-stock
100 mg $1400 In-stock
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  • Biological Activity

  • Protocol

  • Technical Information

  • Purity & Documentation

  • References

Description

PF-3084014 is a reversible, noncompetitive, and selective γ-secretase inhibitor with IC50 of 6.2 nM.

IC50 & Target

IC50: 6.2 nM (γ-secretase)[1]

In Vitro

The IC50 of PF-03084014 for γ-secretase enzyme inhibition in cell-free assay for Aβ production using detergent solubilized membranes derived from HeLa cells is determined to be 6.2 nM. When tested for inhibition of Notch receptor cleavage in cellular assays using HPB-ALL cells that harbor mutations in both the heterodimerization and PEST domains in Notch1, the cell IC50 is determined to be 13.3 nM. PF-03084014 causes a significant increase in caspase-3 activities in HPB-ALL and TALL-1 cells as well as an induction of cleaved PARP and cleaved caspase-3 after a 7-day treatment[1].

In Vivo

PF-03084014 shows robust antitumor activity in this model on 14-day twice daily dosing. Tumor growth inhibition is dose dependent, with maximal tumor growth inhibition of ~92% obtained at high dose levels (150 mg/kg). In tumor growth inhibition studies where mice receive repetitive twice daily dosing for more than a week, PF-03084014 is well tolerated at dose levels below 100 mg/kg as no significant weight loss, morbidity, or mortality is observed. When the dose is increased to 150 mg/kg, however, mice have diarrhea and show weight loss (10-15%) approximately 10 days after compound administration. The body weight of treated animals usually returns to normal if dosing holidays are given, suggesting that the toxicity of PF-03084014 is reversible[1]. In the 7-day repeat dose toxicokinetic (TK) and first 1-month combination repeat dose studies, treatment with Dexamethasone alone and Dexamethasone with PF-03084014 cause moderate to marked body weight loss (-10% to -27%) after 7 days treatment. In the second 1-month combination repeat dose study, a similar magnitude of body weight loss (-10% to 22%) occurs with repeat dosing on the first week or third week of treatment with 100 mg/kg PF-03084014 and 1 mg/kg Dexamethasone. When Dexamethasone is not coadministered with PF-03084014 on the second week of study, increases (4%) in body weight are noted, suggesting that the body weight loss is reversible[2].

References
Preparing Stock Solutions
Concentration Volume Mass 1 mg 5 mg 10 mg
1 mM 2.0423 mL 10.2116 mL 20.4232 mL
5 mM 0.4085 mL 2.0423 mL 4.0846 mL
10 mM 0.2042 mL 1.0212 mL 2.0423 mL
Please refer to the solubility information to select the appropriate solvent.
Kinase Assay
[1]

The source of enzyme for this assay is a crude P2 membrane preparation derived from human HeLa cells. The substrate is affinity-purified recombinant human APP-C100-FLAG peptide produced in Escherichia coli. The membranes are solubilized in detergent (1% CHAPSO) and reactions are assembled and incubated in the presence of various concentration of PF-03084014. Amyloid β peptide (Aβ)1-40 is detected by enzyme-linked immunosorbent assay using β-amyloid, 17-24 (4G8) monoclonal antibody, biotin-labeled and QCB40 rabbit polyclonal antiserum to Aβ40. The IC50 is calculated[1]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Cell Assay
[1]

PF-03084014 is dissolved in DMSO and stored, and then diluted with appropriate media (DMSO 0.1%) before use[1].

Cells are seeded in 96-well plates at 2,000 (Sup-T1, Jurkat, and DND-41) or 10,000 (HPB-ALL or TALL-1) cells/well in growth media supplemented with 10% fetal bovine serum. Serial dilutions of PF-03084014 are done in DMSO, appropriate controls or designated concentrations of PF-03084014 are added to each well, and cells are incubated at 37°C for 7 days (final DMSO content 0.1%). Resazurin at a final concentration of 0.1 mg/mL is added to the cells and plates are incubated for 2 to 4 hours. Fluorescent signals are read as emission at 590 nm after excitation at 560 nm. IC50 values are calculated by using the sigmoidal dose-response (variable slope) in GraphPad Prism[1]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Administration
[1][2]

PF-03084014 is prepared in 0.5% methylcellulose (Mice and Rat)[1][2].

Mice[1]
Athymic female mice (nu/nu, 6-8 weeks) are used. For antitumor efficacy, animals bearing tumors of 150 to 300 mm3 in size are randomly divided into groups that received either vehicle (0.5% methylcellulose) or PF-03084014(150 mg/kg, diluted in vehicle), and dosed by oral gavage. Animal body weight and tumor measurements are obtained every 2 to 3 days. Tumor volume (mm3) is measured with Vernier calipers and calculated. Percent (%) inhibition values are measured on the final day of study for drug-treated compared with vehicle-treated mice and are calculated. For all tumor growth inhibition experiments, 8 to 10 mice per dose group are used. Student's t test is used to determine the P value.
Rat[2]
Sprague-Dawley (SD) rats are useds. In the 7-day repeat dose TK study, 3 male rats per group are orally dosed with either 0.5% methylcellulose vehicle, PF-03084014 at 150 mg/kg/day, PF-03084014 at 150 mg/kg/day coadministered with 1 of the oral Dexamethasone doses (0.25, 1.0, 2.5, or 5.0 mg/kg/day) or Dexamethasone at 5 mg/kg/day and euthanized at 24 hr after the last dosing. The blood collection time points for determining PF-03084014 or Dexamethasone mean systemic plasma concentrations are 1, 2, 4, 7, and 24 hr post dosing. Test article-related findings are determined by assessing changes in clinical signs, pre- and post-dose body weights. Blood samples for assessing systemic exposure of PF-03084014 and Dexamethasone are collected from all treatment groups at various times on days 1 and 7 of the study. Blood samples for hematology evaluation are also collected from all the animals at 24 hr after the last dosing. The mean group changes in hematology parameters for treated rats are expressed as a percentage change. Necropsy is performed 24 hr after the last dose, body weights are recorded, and tissues are collected and submitted for histopathologic examinations. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

References
Molecular Weight

489.64

Formula

C₂₇H₄₁F₂N₅O

CAS No.

1290543-63-3

Storage
Powder -20°C 3 years
  4°C 2 years
In solvent -80°C 6 months
  -20°C 1 month
Shipping

Room temperature in continental US; may vary elsewhere

Solvent & Solubility

10 mM in DMSO

* "<1 mg/mL" means slightly soluble or insoluble. "≥" means soluble, but saturation unknown.

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Product Name:
PF-3084014
Cat. No.:
HY-15185
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