RGFP966

Alias: RGFP966; RGFP-966; RGFP 966

Cat No.:V0272 Purity: ≥98%

COA Product Data Instructions for use SDS

RGFP966 (RGF-P966) is a novel, potent and highly selective inhibitor of HDAC3 (histone deacetylase 3) with potential anticancer activity.

RGFP966 Chemical Structure CAS No.: 1357389-11-7
Product category: HDAC

This product is for research use only, not for human use. We do not sell to patients.

Size	Price	Stock	Qty
1mg
5mg
10mg
25mg
50mg
100mg
250mg
Other Sizes

ADD TO CART CHECKOUT BULK INQUIRY

1-708-310-1919 sales@invivochem.com

Other Forms of RGFP966:

(E,E)-RGFP966

Official Supplier of:

Top Publications Citing lnvivochem Products

Nature 2021, 597(7874):119-125.

Cell 2020,183:1202-1218.e25.

Science Adv 2022: 8, eabm9427.

Nature 597, 119–125 (2021)

Cell Stem Cell 2020,26(6):845-861.e12.

Science Trans Med 2023,15(701):eadd7872.

STTT 2023,8(1):91.

Cell Reports 2023,42(4):112313

Science Trans Med 2023, 15: eadd7872.

Cancer Cell 2021,39(4):529-547.e7.

Cancer Discov 2022,12(4):1106-1127.

Immunity 2023,56(3):620-634.e11.

J Am Chem Soc 2022,144:21831-21836.

Cell 2020,183:1202-1218.e25.

Science Adv 2022:8,eabm9427.

Nature 2021,597(7874):119-125.

Cell 2020,183:1202-1218.e25.

PNAS Nexus 2022,1(3):pgac063.

ACS Nano 2023,17(10):9110-9125.

Biomaterial 2023 Sep16:302:122333.

Purity & Quality Control Documentation

Current batch：

Purity: ≥98%

COA

MSDS

NMR

Instructions

Product Description
Bioactivity & Protocols
Physicochemical Properties
Solubility Data
Calculators
Biological Data

Product Description

RGFP966 (RGF-P966) is a novel, potent and highly selective inhibitor of HDAC3 (histone deacetylase 3) with potential anticancer activity. In a cell-free assay, it inhibits HDAC3 with an IC50 of 80 nM and shows >200-fold selectivity for HDAC3 over other HDACs.

Biological Activity I Assay Protocols (From Reference)

Targets

HDAC3 ( IC50 = 80 nM )

ln Vitro

In vitro activity: RGFP966 is a competitive tight-binding HDAC inhibitor that exhibits slow-on/slow-off behavior. Its IC50 for HDAC3 is 0.08μM, and at concentrations up to 15μM, it does not effectively inhibit any other HDAC.[1] Two CTCL cell lines treated with RGFP966 for 24 hours before western blot analysis showed increased acetylation at H3K9/K14, H3K27, and H4K5, but not at H3K56ac. Due to enhanced apoptosis linked to DNA damage and impeded S phase progression, RGFP966 reduces cell growth in CTCL (cutaneous T cell lymphoma) cell lines. Within the first hour of medication treatment, RGFP966 significantly lowers the DNA replication fork velocity.[2]

ln Vivo

RGFP966 treatment (10 mg/kg)improves object memory's long-term memory. RGFP966 (3 or 10 mg/kg, s.c.) lessens the chance of reinstating cocaine-conditioned place preference and promotes its extinction.[1]

Enzyme Assay

Assays for acetylation are derived from the homogenous fluorescence release method. Following pre-incubation periods ranging from 0 to 3 hours, purified recombinant enzymes are incubated in the standard HDAC buffer with serially diluted inhibitors at the concentrations shown in the figures. After pre-incubation, acetyl-Lys(Ac)-AMC substrate (at 10 μM, corresponding to the Km for both HDAC1 and HDAC3) is added. An hour is given for the reaction to continue. An Tecan M200 96-well plate reader is used to measure the fluorescence emission after an hour of adding the trypsin peptidase developer at a final concentration of 5 mg/ml.

Cell Assay

At 26105 cells/mL, cells are counted and divided into T25 (Corning) flasks. After that, cells are treated once at hour 0 with either DMSO or HDIs. At 0, 24, 48, and 72 hours following treatment, 100 ml aliquots are taken in triplicate from each flask, distributed into a flat bottom 96-well plate, and 10 ml of alamar blue is added to each well. Using the Biotek Synergy MX Microplate Reader, fluorescence is measured after a 4-hour incubation period.

Animal Protocol

Mice: The N171-82Q transgenic mice are kept in their housing and given free access to food and water, as well as a typical 12-hour light/dark cycle beginning at 6:00 a.m. Beginning at 8 weeks of age, mice are given 3 injections per week by subcutaneous injection of RGFP966 (10 or 25 mg/kg) for a duration of 10 weeks. A solution of 75% polyethylene glycol 200/25% sodium acetate (6.25 mM) is used to dissolve RGFP966; mice in the control group were given the same volume of drug vehicle. Weights are recorded twice a week for the body. At eighteen weeks of age, mice are killed by overdosing on isofluorane anesthesia, six hours after the last injection. Brains are taken out, and 4% paraformaldehyde is either intracardially infused, or the cortex and striata are separated out for gene expression experiments.
Rats: The experiment uses thirty-three adult male Sprague Dawley rats weighing between 275 and 350 grams. A posttraining systemic injection of either RGPF966 (10 mg/kg, s.c.) or vehicle (at a volume similar to drug treatment) is administered to each subject right after the daily training session.

References

[1]. Proc Natl Acad Sci U S A . 2013 Feb 12;110(7):2647-52.

[2]. PLoS One . 2013 Jul 22;8(7):e68915.

These protocols are for reference only. InvivoChem does not independently validate these methods.

Physicochemical Properties

Molecular Formula

C21H19FN4O

Molecular Weight

362.4

Exact Mass

362.15

Elemental Analysis

C, 69.60; H, 5.28; F, 5.24; N, 15.46; O, 4.41

CAS #

1357389-11-7

Related CAS #

1396841-57-8

Appearance

White solid powder

SMILES

C1=CC=C(C=C1)/C=C/CN2C=C(C=N2)/C=C/C(=O)NC3=C(C=C(C=C3)F)N

InChi Key

BLVQHYHDYFTPDV-VCABWLAWSA-N

InChi Code

InChI=1S/C21H19FN4O/c22-18-9-10-20(19(23)13-18)25-21(27)11-8-17-14-24-26(15-17)12-4-7-16-5-2-1-3-6-16/h1-11,13-15H,12,23H2,(H,25,27)/b7-4+,11-8+

Chemical Name

(E)-N-(2-amino-4-fluorophenyl)-3-[1-[(E)-3-phenylprop-2-enyl]pyrazol-4-yl]prop-2-enamide

Synonyms

RGFP966; RGFP-966; RGFP 966

HS Tariff Code

2934.99.9001

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)

Solubility Data

Solubility (In Vitro)

DMSO: 50~72 mg/mL (138~198.7 mM)

Water: <1 mg/mL

Ethanol: <1 mg/mL

Solubility (In Vivo)

1% DMSO+30% polyethylene glycol+1% Tween 80: 30mg/mL

(Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.7594 mL	13.7969 mL	27.5938 mL
	5 mM	0.5519 mL	2.7594 mL	5.5188 mL
	10 mM	0.2759 mL	1.3797 mL	2.7594 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.

Calculator

Mass

Concentration

Volume

Molecular Weight*

Calculate Reset

Molarity Calculator allows you to calculate the mass, volume, and/or concentration required for a solution, as detailed below:

Calculate the Mass of a compound required to prepare a solution of known volume and concentration
Calculate the Volume of solution required to dissolve a compound of known mass to a desired concentration
Calculate the Concentration of a solution resulting from a known mass of compound in a specific volume

See Example

An example of molarity calculation using the molarity calculator is shown below:
What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?

Enter 350.26 in the Molecular Weight (MW) box
Enter 10 in the Concentration box and choose the correct unit (mM)
Enter 5 in the Volume box and choose the correct unit (mL)
Click the “Calculate” button
The answer of 17.513 mg appears in the Mass box. In a similar way, you may calculate the volume and concentration.

Concentration (Start)

Volume (Start)

Concentration (End)

Volume (End)

Calculate Reset

Dilution Calculator allows you to calculate how to dilute a stock solution of known concentrations. For example, you may Enter C1, C2 & V2 to calculate V1, as detailed below:

What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:

Enter 10 into the Concentration (Start) box and choose the correct unit (mM)
Enter 25 into the Concentration (End) box and select the correct unit (mM)
Enter 25 into the Volume (End) box and choose the correct unit (mL)
Click the “Calculate” button
The answer of 62.5 μL (0.1 ml) appears in the Volume (Start) box

Molecular formula

Total molecular weight

g/mol

Calculate Reset

Molecular Weight Calculator allows you to calculate the molar mass and elemental composition of a compound, as detailed below:

Note: Chemical formula is case sensitive: C12H18N3O4 c12h18n3o4
Instructions to calculate molar mass (molecular weight) of a chemical compound:

To calculate molar mass of a chemical compound, please enter the chemical/molecular formula and click the “Calculate’ button.

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

Molecular mass (or 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.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

Calculate Reset

Reconstitution Calculator allows you to calculate the volume of solvent required to reconstitute your vial.

Enter the mass of the reagent and the desired reconstitution concentration as well as the correct units
Click the “Calculate” button
The answer appears in the Volume (to add to vial) box

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal to make 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 in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)

% DMSO

% Tween 80

% ddH₂O

Calculate Reset

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.

Note： (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.

Biological Data

HDAC3 inhibition enhances the rate and persistence of extinction of cocaine-CPP.Proc Natl Acad Sci U S A.2013 Feb 12;110(7):2647-52.
HDAC3 inhibition simultaneously reduces drug-seeking behavior and enhances memory.Proc Natl Acad Sci U S A.2013 Feb 12;110(7):2647-52.
HDAC3 inhibition promotes histone acetylation and gene expression during extinction consolidation.Proc Natl Acad Sci U S A.2013 Feb 12;110(7):2647-52.