Molecule

Description

Cancer

Description

Droxinostat (CMH, 5809354) is a selective inhibitor of HDAC3, HDAC6 and HDAC8 with IC50 of 16.9 μM, 2.47μM and 1.46 μM, respectively.

Targets

IC₅₀

In Vitro

Droxinostat is originally identified as a sensitizer of PPC-1 cells to FAS and TRAIL by downregulating the expression of c-Fas-associated death domain-like interleukin-1-converting enzyme-like inhibitory protein (c-FLIP). ^[1]In PPC-1 cells cultured in suspension but not adherent conditions, Droxinostat (20 μM–60 μM) sensitizes cells to anoikis by initially activating caspase 8 with subsequent activation of the mitochondrial pathway. Similarly, Droxinostat also sensitizes other cancer cell lines including PC-3, DU-145, T47D, and OVCAR-3, but not LNCaP or MB-MDA-468, to anoikis or CH-11-induced apoptosis. ^[2]However, the direct targets of Droxinostat remains enigma until recently. It is revealed that in histone deacetylases (HDAC) isoform 1-10, Droxinostat selective inhibits HDAC3, 6, and 8, with IC50 values of 16.9 μM, 2.47 μM, and 1.46 μM, respectively, without inhibiting other HDAC members (IC50 > 20 μM). ^[3]In MCF-7 breast cancer cells, Droxinostat (10 μM–100 μM) sensitizes cells to apoptosis by decreasing c-FLIP_L and c-FLIP_S expression, reducing cell survival, and inducing apoptosis. ^[4]

In Vivo

In SCID mice models, Droxinostat (30 μM)-treated PPC-1 cells results in decreased distant tumor formation than untreated cells. ^[2]

Clinical Trials

Features

Droxinostat is a selective inhibitor of HDAC3, 6, and 8 and acts as a sensitizer to death receptor stimuli.

HDAC Inhibition Assay

HDAC inhibition is assessed using the CycLex HDACs fluorometric assay according to the manufacturer's protocol and using crude nuclear extract from HeLa cells (principally HDAC1 and HDAC2). The relative activity is expressed as (fluorescence intensity of treated samples/fluorescence intensity of controls) × 100%.

Cell Lines

PPC-1 cells

Concentrations

0–100 μM, dissolved in DMSO (final concentration of DMSO is 0.5%)

Incubation Time

24 hours

Methods

PPC-1 cells (1 × 10⁴) are seeded overnight into 96-well flat-bottomed plates in 100 μL of medium containing 2.5% FCS. The next day, Droxinostat is added. CH-11 antibody (100 ng/mL) is then added and the cells are incubated for 24 hours before assessing cell viability by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) dye reduction assay.

Animal Models

Male severe combined immunodeficient (SCID) mice models treated with PPC-1 cells

Formulation

dsRed-PPC-1 cells that treated with Droxinostat (30 μM) or PBS buffer control are either injected via the tail vein or subcutaneously into the hind limbs

Doses

Administration