Nirogacestat (PF-03084014)

Name: Nirogacestat (PF-03084014)
Brand: Selleck Chemicals
SKU: S8018
Rating: 5 (9 reviews)

For research use only.

Catalog No.S8018 Synonyms: PF-3084014

9 publications

Nirogacestat (PF-03084014) Chemical Structure

CAS No. 1290543-63-3

Nirogacestat (PF-03084014, PF-3084014) is a selective gamma-secretase inhibitor with IC50 of 6.2 nM in a cell-free assay. Nirogacestat (PF-03084014, PF-3084014) induces apoptosis. Phase 2.

Selleck's Nirogacestat (PF-03084014) has been cited by 9 publications

Stem Cell Res Ther, 2020, 11(1):258

PubMed: 32586404 ( click the link to review the publication )

Pharm Res, 2020, 37(10):185

PubMed: 32888109 ( click the link to review the publication )

Mol Ther Oncolytics, 2019, 13:58-66

PubMed: 31016228 ( click the link to review the publication )

Front Immunol, 2019, 10:162

PubMed: 30792717 ( click the link to review the publication )

J Cell Physiol, 2019, 234(5):5940-5952

PubMed: 30515785 ( click the link to review the publication )

J Biol Chem, 2019, 294(21):8543-8554

PubMed: 30940724 ( click the link to review the publication )

Exp Cell Res, 2019, 10.1016/j.yexcr.2019.06.015

PubMed: 31211955 ( click the link to review the publication )

Arch Oral Biol, 2019, 100:93-99

PubMed: 30822705 ( click the link to review the publication )

Cell Prolif, 2018, 51(3):e12424

PubMed: 29232766 ( click the link to review the publication )

2 Customer Reviews

The expression of p-EGFR and NICD was assessed by western blot after treatment for 3 d.

Cell Prolif, 2018, 51(3):e12424. Nirogacestat (PF-03084014) purchased from Selleck.

(F) Tumour tissues were further analysed to detect the efficacy of the inhibitors by IHC staining using p-EGFR and Notch1. (4Fa) Notch1 staining was mostly confined on the cell membrane. (4Fc) After Erlotinib treatment, the Notch1 protein was cleaved and translocated to the nucleus. (4Fb) p-EGFR was overexpressed in the control group. (4Fd) Erlotinib treatment inhibited p-EGFR expression. (4Fe-f) PF-03084014 treatment reduced both Notch1 and p-EGFR expression. (4Fg-h) Synthetic therapy reduced the activation of Notch1 and EGFR pathway. Scale bar = 50 μm. *P < .05, **P < .01, ***P < .001

Cell Prolif, 2017, doi: 10.1111/cpr.12424. Nirogacestat (PF-03084014) purchased from Selleck.

Purity & Quality Control

Choose Selective Gamma-secretase Inhibitors

Biological Activity

Description

Targets

γ-secretase ^[1]
(cell-free assay)

6.2 nM

In vitro

PF-03084014 inhibits Notch receptor cleavage in cellular assays using HPB-ALL cells that harbor mutations in both the heterodimerization and PEST domains in Notch1with IC50 of 13.3 nM. PF-03084014 downregulates Notch target genes Hes-1, and cMyc expression in HPB-ALL cells with IC50 of <1 nM and 10 nM, respectively. PF-03084014 inhibits cell growth of a subset of human T-ALL cell lines (HPB-ALL, DND-41, TALL-1,and Sup-T1) through induction of cell cycle arrest and apoptosis with IC50s of 30-100 nM. ^[1] PF-03084014 reduces proliferation of HUVECs with IC50 of 0.5 μM, and decreases the lumen formation with an IC50 value of 50 nM. PF-03084014 (1 μM) has no antiproliferative effect in MX1 cells; however, it inhibits migration by 95%. ^[2]

Assay

Methods	Test Index	PMID
Western blot	N1ICD / Hes-1 / Hey-1 / p-MEK / MEK / c-PARP ; PubMed: 23402814 Cancer Lett Immunobloting of lysates from Panc265 orthotopic experiment demonstrating that PF-03084014 and combination of GEM plus PF-03084014 strongly inhibit N1ICD and the expression of Notch targets Hes-1 and Hey-1. Cleaved-PARP (c-PARP) levels were elevated in GEM and GEM plus PF-03084014 treatment, indicating higher apoptosis compared to Control or PF-03084014 treatment groups. While the treatments could not modulate MEK1/2 protein expression, PF-03084014 and combination of GEM plus PF-03084014 treatment inhibit MEK1/2 phosphorylation. Numbers on the left side of the blots indicate the molecular weight of the proteins in kilodaltons.	23402814

In vivo

PF-03084014 orally administrated in a single dose of 200 mg/kg, causes maximal NICD inhibition for ∼80% in xenograft HPB-ALL tumors. PF-03084014 shows robust antitumor activity in this mode with a maximal tumor growth inhibition of ∼ 92% at dose of 150 mg/kg, accompanied by a significant reduction of NICD/Notch1, tumor mitotic index (Ki67), and apoptosis (activated caspase-3) staining. ^[1] PF-03084014 (120 mg/kg) induces apoptosis, antiproliferation, reduces tumor cell self-renewal ability, impaires tumor vasculature, and decreases metastasis activity in breast cancer HCC1599 tumor-bearing mice. PF-03084014 treatment displays significant antitumor activity in various types of the breast xenograft models with TGI value of at least 50%. ^[2]

Protocol

Kinase Assay:^[3]	- Collapse γ-secretase assay: A DNA fragment encoding amino acids 596 - 695 of the 695-aa isoform of APP (APP695) and the Flag sequence (DYKDDDDK) at the C terminus is generated by PCR amplification with suitably designed oligonucleotides and the APP695 cDNA. The Met that serves as the translation start site is residue 596 of APP695 (the P1 residue with respect to theβ-secretase cleavage site). This DNA fragment is inserted into the prokaryotic expression vector pET2-21b. The recombinant protein, C100Flag, is overproduced in Escherichia coli [strain BL21(DE3)] and purified by Mono-Q column chromatography. C100Flag (1.7 μM) is incubated with cell membranes (0.5 mg/mL) in the presence of CHAPSO, CHAPS (3-[(3-cholamidopropyl)dim-ethylammonio]-1-propanesulfonate), or Triton X-100 (0, 0.125, 0.25, 0.5, or 1%) in buffer B (50 mM Pipes, pH 7.0y 5mM MgCl₂/5 mM CaCl₂/150 mM KCl) at 37°C. The reactions are stopped by adding RIPA (150 mM NaCl/1.0% NP-40/0.5% sodium deoxycholatey 0.1% SDS/50 mM Tris HCl, pH 8.0) and boiling for 5 min. The samples ae centrifuged and the supernatant solutions are assayed for the Aβ peptides by ECL. The Aβ40- and Aβ42-related products from γ-secretase-mediated processing of C100Flag possess a Met at the N terminus and are thus defined as M-Aβ40 and M-Aβ42, respectively. Likewise, supernatant solution (0.125 mg/mL) from CHAPSO-extracted HeLa cell membranes (solubilized γ-secretase) is incubated with C100Flag (1.7 μM) in buffer B containing 0.25% CHAPSO and subsequently assayed for M-Aβ40 and M-Aβ42 by using ECL.
Cell Research:^[1]	- Collapse Cell lines: Human T-ALL cell lines HPB-ALL Concentrations: ~1 μM Incubation Time: 7 days Method: Cells are seeded in 96-well plates at 10,000 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℃ 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. (Only for Reference)
Animal Research:^[1]	- Collapse Animal Models: Human T-cell acute lymphoblastic leukemia xenografts HPB-ALL Dosages: 150 mg/kg Administration: p.o. twice daily (Only for Reference)

References

Solubility (25°C)

In vitro	DMSO	97 mg/mL warmed (198.1 mM)
	Water	Insoluble
	Ethanol	'97 mg/mL warmed

* 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 Information Download Nirogacestat (PF-03084014) SDF

Molecular Weight	489.64
Formula	C₂₇H₄₁F₂N₅O
CAS No.	1290543-63-3
Storage	3 years-20°C powder 2 years-80°C in solvent
Synonyms	PF-3084014
Smiles	CCCC(C(=O)NC1=CN(C=N1)C(C)(C)CNCC(C)(C)C)NC2CCC3=C(C2)C(=CC(=C3)F)F

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

Dosage

mg/kg

Average weight of animals

Dosing volume per animal

Number of animals

Step 2: Enter the in vivo formulation (Different batches have different solubility ratios, please contact Selleck to provide you with the correct ratio)

% DMSO+ % + % Tween 80+ % ddH₂O

Calculate Reset

Calculation results:

Working concentration： mg/ml；

Method for preparing DMSO master liquid: ： mg drug pre-dissolved in μL DMSO (Master liquid 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 DMSO master liquid, next addμL PEG300， mix and clarify, next addμL Tween 80，mix and clarify, next add μL ddH₂O，mix and clarify.

Note：1.Please make sure the liquid is clear before adding the next solvent.
2.Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.

Bio Calculators

Molarity Calculator

Calculate the mass, volume or concentration required for a solution. The Selleck molarity calculator is based on the following equation:

Mass (mg) = Concentration (mM) × Volume (mL) × Molecular Weight (g/mol)

*When preparing stock solutions, please always use the batch-specific molecular weight of the product found on the via label and MSDS / COA (available on product pages).

Dilution Calculator

Calculate the dilution required to prepare a stock solution. The Selleck dilution calculator is based on the following equation:

Concentration _(start) x Volume _(start) = Concentration _(final) x Volume _(final)

This equation is commonly abbreviated as: C1V1 = C2V2 ( Input Output )

* When preparing stock solutions always use the batch-specific molecular weight of the product found on the vial label and MSDS / COA (available online).

The Serial Dilution Calculator Equation

Serial Dilutions
Concertration (start):
Dilution-folds:

Computed Result

C1=C0/X	C1:	LOG(C1):
C2=C1/X	C2:	LOG(C2):
C3=C2/X	C3:	LOG(C3):
C4=C3/X	C4:	LOG(C4):
C5=C4/X	C5:	LOG(C5):
C6=C5/X	C6:	LOG(C6):
C7=C6/X	C7:	LOG(C7):
C8=C7/X	C8:	LOG(C8):

Molecular Weight Calculator

Enter the chemical formula of a compound to calculate its molar mass and elemental composition:

Tip: Chemical formula is case sensitive. C10H16N2O2 c10h16n2o2

Instructions to calculate molar mass (molecular weight) of a chemical compound:

To calculate molar mass of a chemical compound, please enter its chemical formula and click 'Calculate'.

Definitions of molecular mass, molecular weight, molar mass and molar weight:

Molecular mass (molecular weight) is the mass of one molecule of a substance and is expressed in the unified atomic mass units (u). (1 u is equal to 1/12 the mass of one atom of carbon-12)
Molar mass (molar weight) is the mass of one mole of a substance and is expressed in g/mol.

Molarity Calculator

Clinical Trial Information (data from https://clinicaltrials.gov, updated on 2020-09-01)

NCT Number	Recruitment	interventions	Conditions	Sponsor/Collaborators	Start Date	Phases
NCT02338531	Withdrawn	Drug: PF-03084014\|Procedure: Breast cancer surgery	Breast Cancer	Jules Bordet Institute	June 2015	Phase 2
NCT02299635	Terminated	Drug: PF-03084014	Triple Negative Breast Neoplasms	Pfizer	February 3 2015	Phase 2
NCT02109445	Terminated	Drug: PF-03084014\|Drug: Gemcitabine\|Drug: Nab-paclitaxel	Metastatic Cancer Pancreas	Pfizer\|Academic GI Cancer Consortium (AGICC)	September 3 2014	Phase 2
NCT01876251	Terminated	Drug: PF-03084014\|Drug: Docetaxel	Breast Cancer Metastatic	Pfizer	November 4 2013	Phase 1

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.

Handling Instructions

Tel: +1-832-582-8158 Ext:3

If you have any other enquiries, please leave a message.

Gamma-secretase Signaling Pathway Map

Related Gamma-secretase Products

AZD3839 ^New

AZD3839 is a potent and selective BACE1 inhibitor with K_i of 26.1 nM, about 14-fold selectivity over BACE2. Phase 1.
Xanthohumol ^New

Xanthohumol, a prenylated chalcone from hop, inhibits COX-1 and COX-2 activity and shows chemopreventive effects. Xanthohumol inhibits diacylglycerol acyltransferase 1 (DGAT1) and DGAT2 with both IC50 of 40 μM. Xanthohumol is also a potent antiviral agent against a series of DNA and RNA viruses. Xanthohumol induces growth inhibition and apoptosis in cancer cells. Phase 1.
A-438079 HCl ^New

A-438079 HCl is a potent, and selective P2X7 receptor antagonist with pIC50 of 6.9.
Avagacestat (BMS-708163)

Avagacestat (BMS-708163) is a potent, selective, orally bioavailable γ-secretase inhibitor of Aβ40 and Aβ42 with IC50 of 0.3 nM and 0.27 nM, demonstrating a 193-fold selectivity against Notch. Phase 2.

Features:Appears to be more “notch sparing” than semagacestat (LY450139).
RO4929097

RO4929097 is a γ secretase inhibitor with IC50 of 4 nM in a cell-free assay, inhibiting cellular processing of Aβ40 and Notch with EC50 of 14 nM and 5 nM, respectively. Phase 2.
Semagacestat (LY450139)

Semagacestat (LY450139) is a γ-secretase blocker for Aβ42, Aβ40 and Aβ38 with IC50 of 10.9 nM, 12.1 nM and 12.0 nM, also inhibits Notch signaling with IC50 of 14.1 nM in H4 human glioma cell. Phase 3.

Features:The best characterized γ-secretase inhibitor that has reached the clinic.
DAPT (GSI-IX)

DAPT (GSI-IX, LY-374973) is a novel γ-secretase inhibitor, which inhibits Aβ production with IC50 of 20 nM in HEK 293 cells. DAPT enhances the apoptosis of human tongue carcinoma cells and regulates autophagy.
MK-0752

MK-0752 is a moderately potent γ-secretase inhibitor, which reduces Aβ40 production with IC50 of 5 nM. Phase 1/2.

Features:A moderately potent γ-secretase inhibitor.
LY411575

LY411575 is a potent γ-secretase inhibitor with IC50 of 0.078 nM/0.082 nM (membrane/cell-based), also inhibits Notch cleavage with IC50 of 0.39 nM in APP or NΔE expressing HEK293 cells. LY411575 induces apoptosis.
Dibenzazepine (YO-01027)

Dibenzazepine (YO-01027) is a dipeptidic γ-secretase inhibitor with IC50 of 2.6 nM and 2.9 nM in cell-free assays for APPL and Notch cleavage, respectively.

Choose Your Country or Region

By Signaling Pathways

By product type

Nirogacestat (PF-03084014)