PCI-34051 950762-95-5_HDAC_DNA Damage_Signaling Pathways_Products

PCI-34051 is a novel, potent and specific inhibitor of histone deacetylase 8 (HDAC8) with potential anticancer activity. It exhibits >200-fold selectivity for HDAC8 over HDAC1 and 6, and 1000-fold higher selectivity over HDAC2/3/10. In a cell-free assay, it inhibits HDAC8 with an IC50 of 10 nM.

Physicochemical Properties

Molecular Formula

C17H16N2O3

Molecular Weight

296.32

Exact Mass

296.116

Elemental Analysis

C, 68.91; H, 5.44; N, 9.45; O, 16.20

CAS #

950762-95-5

Related CAS #

950762-95-5

PubChem CID

24753719

Appearance

White to gray solid powder

Density

1.3±0.1 g/cm3

Index of Refraction

1.621

LogP

2.2

Hydrogen Bond Donor Count

Hydrogen Bond Acceptor Count

Rotatable Bond Count

Heavy Atom Count

Complexity

382

Defined Atom Stereocenter Count

SMILES

O=C(C1C=C2C(C=CN2CC2C=CC(OC)=CC=2)=CC=1)NO

InChi Key

AJRGHIGYPXNABY-UHFFFAOYSA-N

InChi Code

InChI=1S/C17H16N2O3/c1-22-15-6-2-12(3-7-15)11-19-9-8-13-4-5-14(10-16(13)19)17(20)18-21/h2-10,21H,11H2,1H3,(H,18,20)

Chemical Name

N-hydroxy-1-[(4-methoxyphenyl)methyl]indole-6-carboxamide

Synonyms

PCI-34051; PCI 34051; PCI34051

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)

Biological Activity

Targets

HDAC8 ( IC50 = 10 nM ); HDAC6 ( IC50 = 2.9 μM ); HDAC1 ( IC50 = 4 μM ); HDAC10 ( IC50 = 13 μM )

ln Vitro

In vitro activity: PCI-34051 has a promising potency for HDAC8 with a Ki of 10 nM. PCI-34051 exhibits a high degree of selectivity (roughly five times) for HDAC8 in comparison to other class I HDACs, such as HDAC1. More than 200-fold selectivity over HDAC1 and HDAC6, as well as more than 1000-fold selectivity over HDAC2, HDAC3, and HDAC10, are revealed by PCI-34051. OVCAR-3 ovarian tumor line is inhibited by PCI-34051 with a GI50 of 6 μM and 15% cell death. In the sensitive cell lines treated with PCI-34051 at concentrations less than 25 μM at 24 hours or earlier, no significant histone acetylation or tubulin acetylation is seen. In cell lines that are exclusively derived from T-cell malignancies, PCI-34051 selectively induces cytotoxicity. PCI-34051 causes apoptosis in a caspase-dependent manner. There is an increase in activity from 12 to 24 to 48 hours after treatment with 5 μM PCI-34051, which is consistent with the higher levels of caspase activity at this timepoint and is another sign of apoptosis. A characteristic effect of the extrinsic apoptotic pathway, Bid cleavage, is not stimulated by PCI-34051. PLCγ1-deficient J.gamma1 line shows a significant reduction in the amount of PCI-34051-induced apoptosis, whereas P116 and J.RT3-T.5 are sensitive to PCI-34051. Moreover, the apoptosis caused by PCI-34051 is significantly impacted by steady-state calcium levels. PCI-34051 causes the release of cytochrome c from the mitochondria.[1]

ln Vivo

In order to lessen airway remodeling in asthma, PCI-34051 and dexamethasone administration lowers eosinophilic inflammation and airway hyperresponsiveness.

Enzyme Assay

Measurements are conducted in a reaction volume of 100 μL using 96-well assay plates in a fluorescence plate reader for PCI-34051 characterization. for every isozyme. The HDAC protein is combined with PCI-34051 at different concentrations and incubated for 15 minutes in reaction buffer (50 mM HEPES, 100 mM KCl, 0.001% Tween-20, 5% dimethyl sulfoxide, pH7.4) supplemented with bovine serum albumin at concentrations of 0-0.05%. The reaction is started by adding acetyl-gly-Ala-(N-acetyl-Lys)-amino-4-methylcoumarin at a final concentration of 25–100 μM and trysin at a final concentration of 50 nM. The fluorescence is measured over a 30-minute period with an excitation wavelength of 335 nm and a detection wavelength of 460 nm, following a 30-minute lag time. The reaction rate is determined by measuring the increase in fluorescence with time.

Cell Assay

Human umbilical vein endothelial cells and tumor cell lines are cultured for at least two doubling times, and growth is assessed using an Alamar Blue fluorometric cell proliferation assay, per the manufacturer's recommendation, at the conclusion of PCI-34051 exposure. In 96-well plates, PCI-34051 is tested in triplicate wells. A logistic equation with four parameters is used to estimate the concentration needed to inhibit cell growth by 50% (GI50) and the 95% confidence intervals from nonlinear regression.

Animal Protocol

Mice: There is an asthma mouse model used. In short, the mice used are 72 healthy female BALB/C mice that weigh between 18 and 22 g and are 6 to 8 weeks old. The animals are kept apart and given unlimited access to normal food and water in a pathogen-free environment. The week before the experiment starts, the animals are housed. Six treatment groups consist of the following mice: PCI-34051, Givinostat, Dexamethasone, Tubastatin A HCl, normal control, and simple asthma. Ovabumin (OVA, 20 μg) and aluminum hydroxide gel (2 mg) are used to sensitize the mice in the final five groups on the first, eighth, and fifteenth days. Seven days following the previous sensitization, an ultrasonic atomizing device (3 mL/min for 30 min, three times/week for 8 weeks) is used to atomize OVA (20 mg/mL). Intraperitoneal injections of Dexamethasone (2.0 mg/kg), TSA (0.5 mg/kg), PCI-34051 (0.5 mg/kg), and Givinostat (0.5 mg/kg) are given half an hour prior to excitation. OVA is substituted with normal saline in the normal control group.

References

[1]. A novel histone deacetylase 8 (HDAC8)-specific inhibitor PCI-34051 induces apoptosis in T-cell lymphomas. Leukemia. 2008 May;22(5):1026-34.

[2]. Therapeutic effects of histone deacetylase inhibitors in a murine asthma model. Inflamm Res. 2016 Dec;65(12):995-1008.

Additional Infomation

N-hydroxy-1-[(4-methoxyphenyl)methyl]-6-indolecarboxamide is an indolecarboxamide.

Solubility Data

Solubility (In Vitro)

DMSO: ~59 mg/mL (~199.1 mM)

Water: <1 mg/mL

Ethanol: <1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (8.44 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 (8.44 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.

Solubility in Formulation 3: ≥ 2.5 mg/mL (8.44 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly.

Solubility in Formulation 4: 30% PEG400+0.5% Tween80+5% propylene glycol: 30mg/mL

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

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	3.3747 mL	16.8737 mL	33.7473 mL
	5 mM	0.6749 mL	3.3747 mL	6.7495 mL
	10 mM	0.3375 mL	1.6874 mL	3.3747 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.

PeptideDB

PCI-34051 950762-95-5

CAS No.: 950762-95-5

Physicochemical Properties

Biological Activity

Solubility Data