HPOB 1429651-50-2_HDAC_DNA Damage_Signaling Pathways_Products

HPOB is a novel, potent and selective inhibitor of histone deacetylase 6 (HDAC6) with potential anticancer activity. It exhibits >30-fold selectivity for HDAC6 over other HDAC isoforms and inhibits HDAC6 with an IC50 of 56 nM.

Physicochemical Properties

Molecular Formula

C17H18N2O4

Molecular Weight

314.34

Exact Mass

314.126

Elemental Analysis

C, 64.96; H, 5.77; N, 8.91; O, 20.36

CAS #

1429651-50-2

Related CAS #

1429651-50-2

PubChem CID

71532921

Appearance

Off-white to pink solid powder

Density

1.3±0.1 g/cm3

Index of Refraction

1.649

LogP

-0.05

Hydrogen Bond Donor Count

Hydrogen Bond Acceptor Count

Rotatable Bond Count

Heavy Atom Count

Complexity

388

Defined Atom Stereocenter Count

SMILES

O=C(N(CCO)C1=CC=CC=C1)CC2=CC=C(C(NO)=O)C=C2

InChi Key

RFAZNTABYJYOAR-UHFFFAOYSA-N

InChi Code

InChI=1S/C17H18N2O4/c20-11-10-19(15-4-2-1-3-5-15)16(21)12-13-6-8-14(9-7-13)17(22)18-23/h1-9,20,23H,10-12H2,(H,18,22)

Chemical Name

N-hydroxy-4-[2-[N-(2-hydroxyethyl)anilino]-2-oxoethyl]benzamide

Synonyms

HPOB

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

HDAC6 ( IC50 = 0.056 μM ); HDAC3/NCOR2 ( IC50 = 1.7 μM ); HDAC8 ( IC50 = 2.8 μM ); HDAC1 ( IC50 = 2.9 μM ); HDAC10 ( IC50 = 3.0 μM ); HDAC2 ( IC50 = 4.4 μM )

ln Vitro

HPOB (8, 16, or 32 μM; 72 hours) inhibits the growth of normal or transformed cells, but not their viability[1].
HPOB causes acetylated α-tubulin and acetylated peroxiredoxin, substrates of HDAC6, to accumulate in both normal (HFS) and transformed (LNCAP, U87, and A549) cells, but not acetylated histones. HPOB increases the death of transformed cells (LNCAP, U87, and A549 cells) induced by etoposide, doxorubicin, and SAHA, but not normal cell death[1].
Clever PARP (a sign of apoptosis) was found to be upregulated in LNCaP cells treated with HPOB and etoposide as well. The accumulation of γH2AX in LNCaP cells was indicative of increased DNA damage accumulation when HPOB and etoposide were combined, as opposed to when etoposide was used alone[1].
HPOB reduces the damage caused by corticosterone in rat adrenal pheochromocytoma PC12 cells by blocking the intrinsic apoptotic pathway and mitochondrial GR translocation[2].

ln Vivo

HPOB (300 mg/kg; i.p.; daily for 18 days) and SAHA (50 mg/kg) inhibits the development of CWR22 tumors that have already grown[1].

Enzyme Assay

The fluorogenic release of 7-amino-4-methylcoumarin from substrate upon deacetylase enzymatic activity is used to detect the in vitro activities of the 11 recombinant human zinc-dependent HDAC enzymes. Ten percent DMSO in HDAC assay buffer is used to prepare a series of dilutions of the unique HDAC6 compound, tubacin, and SAHA. Five microliters of each dilution is then added to a 50 microliter reaction, ensuring that the final concentration of DMSO is one percent in each reaction. The enzymatic reactions are carried out in duplicate in a 50-μL mixture containing an HDAC substrate, an HDAC enzyme, an HDAC assay buffer, 5 μg BSA, and a test compound for 30 minutes at 37 °C. Following the enzymatic reactions, 50 μL of 2× HDAC developer is added to every well, and the plate is left to incubate for a further fifteen minutes at room temperature. A Synergy microplate reader is used to measure the fluorescence intensity at an excitation of 360 nm and an emission of 460 nm. The assays include positive controls (SAHA, a known HDAC inhibitor) and negative controls (no enzyme, no inhibitor, or medication with no HDAC inhibition activity). The drug concentration that causes a 50% decrease in HDAC activity when compared to the control is known as the IC50.

Cell Assay

The prescribed doses of HPOB are cultivated in normal (HFS) and transformed (LNCaP, A549, and U87) cells for a duration of 72 hours. A positive control is five micromolars of SAHA. Prism 5 was utilized to construct the graphs.

Animal Protocol

Dissolved in DMSO; 300 mg/kg daily; i.p. injection.

Mice bearing CWR22 human prostate cancer xenografts

References

[1]. Proc Natl Acad Sci U S A . 2013 Sep 24;110(39):15704-9.

Solubility Data

Solubility (In Vitro)

DMSO: 50~62 mg/mL (159.1~197.2 mM)

Water: <1 mg/mL

Ethanol: ~38 mg/mL warmed (120.9mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.95 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 (7.95 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 (7.95 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.

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

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	3.1813 mL	15.9063 mL	31.8127 mL
	5 mM	0.6363 mL	3.1813 mL	6.3625 mL
	10 mM	0.3181 mL	1.5906 mL	3.1813 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

HPOB 1429651-50-2

CAS No.: 1429651-50-2

Physicochemical Properties

Biological Activity

Solubility Data