Name: RO4929097 (RG-4733)
Brand: Selleck Chemicals
SKU: S1575
Availability: InStock
Rating: 5 (125 reviews)

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Quality Control

Batch: Purity: 99.77%

99.77

Related Targets	HDAC Caspase Proteasome MMP HCV Protease Cysteine Protease Tyrosinase DPP HIV Protease Serine Protease
Other Secretase Inhibitors	DAPT LY411575 Nirogacestat (PF-03084014) Dibenzazepine (YO-01027) MK-0752 Semagacestat (LY450139) Avagacestat (BMS-708163) MDL-28170 L-685,458 Itanapraced (CHF 5074)

Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Incubation Time	Formulation	Activity Description	PMID
U87	Cell Viability Assay	30 μM	4 d		decreases cell viability	24495907
U87 R1	Cell Viability Assay	30 μM	4 d		decreases cell viability	24495907
MGG4	Cell Viability Assay	30 μM	4 d		decreases cell viability	24495907
MGG6	Cell Viability Assay	30 μM	4 d		decreases cell viability	24495907
MGG8	Cell Viability Assay	30 μM	4 d		decreases cell viability	24495907
MGG23	Cell Viability Assay	30 μM	4 d		decreases cell viability	24495907
SUM149	Cell Viability Assay	0-10 μM	72 h		inhibits cell growth dose dependently	22547109
Sum190	Cell Viability Assay	0-10 μM	72 h		inhibits cell growth dose dependently	22547109
WM35	Function Assay	10 uM	24 h	DMSO	decreases the levels of NOTCH downstream target HES1	21980408
WM98.1	Function Assay	10 uM	24 h	DMSO	decreases the levels of NOTCH downstream target HES1	21980408
WM115	Function Assay	10 uM	24 h	DMSO	decreases the levels of NOTCH downstream target HES1	21980408
WM983A	Function Assay	10 uM	24 h	DMSO	decreases the levels of NOTCH downstream target HES1	21980408
WM3248	Function Assay	10 uM	24 h	DMSO	decreases the levels of NOTCH downstream target HES1	21980408
WM35	Growth Inhibition Assay	10 uM	0-18 d	DMSO	inhibits cell growth time dependently	21980408
WM98.1	Growth Inhibition Assay	10 uM	0-18 d	DMSO	inhibits cell growth time dependently	21980408
WM115	Growth Inhibition Assay	10 uM	0-18 d	DMSO	inhibits cell growth time dependently	21980408
WM983A	Growth Inhibition Assay	10 uM	0-18 d	DMSO	inhibits cell growth time dependently	21980408
WM3248	Growth Inhibition Assay	10 uM	0-18 d	DMSO	inhibits cell growth time dependently	21980408
A673	qHTS assay					29435139
DAOY	qHTS assay					29435139
BT-37	qHTS assay					29435139
BT-12	qHTS assay					29435139
OHS-50	qHTS assay					29435139
SJ-GBM2	qHTS assay					29435139
SK-N-MC	qHTS assay					29435139
NB-EBc1	qHTS assay					29435139
LAN-5	qHTS assay					29435139
Click to View More Cell Line Experimental Data

Solubility

In vitro
Batch:

DMSO : 94 mg/mL (200.25 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Ethanol : 50 mg/mL

Water : Insoluble

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
Mass value Mass unit	Concentration value Concentration unit	Volume value Volume unit	Molecular weight (g/mol)

Dilution Calculator Molecular Weight Calculator

In vivo batch selection In vivo Batch:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 95%saline Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	0.200mg/ml (0.43mM)	Taking the 1 mL working solution as an example, add 50 μL of 4 mg/ml clear DMSO stock solution to 950 μL of saline and mix evenly. The mixed solution should be used immediately for optimal results.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	4.700mg/ml (10.01mM)	Taking the 1 mL working solution as an example, add 50 μL of 94 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.

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: g

Dosing volume per animal: μL

Number of animals:

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO

%

% Tween 80

% ddH₂O

% DMSO

+

%

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 μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, 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.

Chemical Information, Storage & Stability

Molecular Weight	469.4	Formula	C₂₂H₂₀F₅N₃O₃	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	847925-91-1	Download SDF		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
Synonyms	RG-4733			Smiles	CC(C)(C(=O)NCC(C(F)(F)F)(F)F)C(=O)NC1C2=CC=CC=C2C3=CC=CC=C3NC1=O

Mechanism of Action

Targets/IC₅₀/Ki	γ secretase (Cell-free assay) 4 nM Notch (Cell-free assay) 5 nM Aβ40 (Cell-free assay) 14 nM
In vitro	RO4929097 decreases the amount of Aβ peptides secreted into the culture medium in HEK293 cells with EC50 of 14 nM. This compound strongly inhibits Notch processing with EC50 of 5 nM in the Notch cell-based reporter assay. The potency of this chemical in cell-free and cellular assays is in the low nanomolar range with >100-fold selectivity observed with respect to 75 other proteins of various types including receptors, ion channels, and enzymes (CEREP panel). After 5 days of treatment, it reduces the production of ICN in the human NSCLC A549 cells inducing a flattened and less transformed tumor cell phenotype in tissue culture. It blocks Notch processing in human non-small cell lung carcinoma cells and decreases expression of the Notch transcriptional target gene Hes1. Treatment with this compound reveals a two- to threefold decrease in the expression of direct Notch target genes, Hes1, Hey1, and Heyl in SUM149 and a 3.5- to eightfold decrease in expression in SUM190 cells. It modestly inhibits the growth of SUM149 cells in a dose-dependent manner. At a concentration of 1 μM of this chemical, growth inhibition is 20 % for SUM149 and 10 % for SUM190 cells, relative to vehicle-treated controls. It decreases the production of inflammatory cytokines by T-cells. Furthermore, with this treatment, there is a shift in favor of TH2 over TH1 cytokines. In addition, T-cell activation induced IL-6 production would be increased with this compound. Upon this treatment, the selected melanoma cell lines reveals downregulation of NOTCH downstream effector HES1. A decrease in the amount of melanospheres formed upon this treatment in primary melanoma cell lines is detected.
Kinase Assay	In vitro potency assays
Kinase Assay	After RO4929097 is used, the Aβ peptides are measured by ECL assays using a variety of anti-Aβ antibodies and an Origen 1.5 Analyzer. The 4G8 murine mAb binds an epitope in the Aβ peptide (within amino acids 18–21) that is immediately distal to the α-secretase cleavage site. The G2–10 murine mAb binds the C terminus that is exposed after γ-secretase-mediated cleavage to generate amino acid 40 of the Aβ40 peptide. The FCA3542 rabbit antibody binds the C terminus that is exposed after γ-secretase-mediated cleavage to generate amino acid 42 of the Aβ42 peptide. The 4G8 mAb is biotinylated with biotin-LC-sulfo-N-hydroxysuccinimide-ester. The G2–10 and FCA3542 antibodies are ruthenylated with TAG-N-hydroxysuccinimide ester. Aβ(x-40) is detected with biotinylated 4G8 and ruthenylated G2–10. Aβ(x-42) is detected with biotinylated 4G8 and ruthenylated FCA3542.
In vivo	Oral injection of 3 to 60 mg/kg RO4929097 once daily or twice daily to nude mice bearing A549 NSCLC xenografts for either 7, 14, or 21 days of a 21-day schedule results in significant tumor growth inhibition compared with vehicle-treated animals. The tumor growth inhibition values ranges from 66% to 91%. When mice are treated with 60 mg/kg of this compound twice daily with the 7+/14- schedule, treatment initially arouses regression of established A549 tumors. At the end of the 21-day cycle (day 47), tumor growth prevention is still 91% compared with vehicle control mice. Inhibition of tumor growth remains prolonged and sustained up to 34 days post-treatment (day 67). On day 67, these mice are retreated with the same dose of this chemical for a second cycle (7 days) until day 74. Importantly, the antitumor effects are sustained after dosing is completed. This compound leads to reduced expression of genes associated with angiogenesis in A549 xenograft model. In contrast, the RO4929097-resistant H460a xenograft displays little change in expression of these genes, underscoring the in vivo anti-angiogenesis mechanism of action of this agent. For IL6 and IL8 overexpressing tumors, it no longer impacts angiogenesis or the infiltration of tumor associated fibroblasts.
References	[1] https://pubmed.ncbi.nlm.nih.gov/19773430/ [2] https://pubmed.ncbi.nlm.nih.gov/22547109/ [3] https://pubmed.ncbi.nlm.nih.gov/21980408/ [4] https://pubmed.ncbi.nlm.nih.gov/21315665/ [5] https://pubmed.ncbi.nlm.nih.gov/10801983/

Applications

Methods	Biomarkers	Images	PMID
Western blot	Hey1 Snail / N-cadherin / Twist / E-cadherin Akt / p-Akt / Notch / IGF1R / FBXW7 NICD / Hes1 / Hes3 / Hes5		29899322
Growth inhibition assay	Cell viability		30669546

Tech Support

Handling Instructions

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

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Frequently Asked Questions

Question 1:
How about its half-life?

Answer:
Its half-life is about 20 hours based on the following paper: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3869895/

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RO4929097 (RG-4733) γ-Secretase inhibitor

Chemical Structure

Molecular Weight: 469.4