Inhibitory activities

Recombinant HDAC activity assays were done by Reaction Biology Corporation. In all cases, full-length HDAC proteins were expressed using baculovirus-infected Sf9 cells. In addition, HDAC3 was coexpressed as a complex with human NCOR2. For assessing activity of HDAC1-containing cellular complexes, immunoprecipitated HDAC1 complexes were incubated with an [3H]acetyllabeled fragment of histone H4 peptide [biotin-(6-aminohexanoic)Gly-Ala-(acetyl[3H])Lys-Arg-His-Arg-Lys-Val-NH2]. Equal amounts of HDAC1 were immunoprecipitated as indicated by Western blot analysis. HDAC1 activity results are presented as mean ± SD of three independent experiments on a single lysate.

Cell lines

Non-small cell lung carcinoma (NSCLC) cell lines; hematologic cell lines; human tumor cells; human A2780 ovarian carcinoma cells.

Preparation method

Soluble in DMSO >10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reaction Conditions

cell proliferation: 72 h, apoptosis assays: 24, 48, and 96 h.

Applications

In all lung, breast, colon, prostate, brain, and ovarian tumor cell lines tested, JNJ-26481585 inhibits cell proliferation with IC50 values of 3.1-246 nM. In A2780 ovarian tumor cells, JNJ-26481585 (3-300 nM) significantly and dose-dependently increases the amount of cells positive for Annexin V, which indicates apoptosis.

Animal models

Male athymic nu/nu CD-1 mice injected s.c. with human A2780-p21waf1,cip1 ZsGreen ovarian tumors cells

Dosage form

10 mg/kg i.p. and 40 mg/kg p.o.; once daily for 3 d.

Preparation method

Formulated at 2 mg/mL in 20% hydroxypropyl-β-cyclodextrin (final pH 8.7).

Applications

In the HDAC1-responsive A2780 ovarian tumor screening model, JNJ-26481585 induces a bright and intense fluorescence, which is not uniformly distributed throughout the tissue and fully predicts tumor growth inhibition. Also, JNJ-26481585 induces potent H3 acetylation in the tumor tissue. In HCT116 colon xenografts, JNJ-26481585 (once daily, 10 mg/kg i.p.) for 14 days inhibits tumor volume by 76% and induces H3 acetylation.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

JNJ-26481585 is a novel histone deacetylase (HDAC) inhibitor that exerts a strong potency towards class I HDAC (including HDAC1, HDAC2 and HDAC3), with values of inhibition constant IC₅₀of 0.11, 0.33 and 4.8 nmol/L respectively, leading to strong anti-proliferative activities (IC₅₀ranging from 3.1 to 246 nmol/L) against a broad range of cancer cell lines including lung, breast, colon, prostate, brain and overian cancer cell lines. Recent study results have shown that JNJ-26481585 strongly induce the HDAC1-suppressed p21^waf1,cip1promoter in vivo, histone H3 acetylation in tumor tissue and apoptosis in human colon cancer cell lines (both APC wild-type and mutant) in vitro.

Reference

Arts J, King P, Marien A, Floren W, Belien A, Janssen L, Pilatte I, Roux B, Decrane L, Gilissen R, Hickson I, Vreys V, Cox E, Bol K, Talloen W, Goris I, Andries L, Du Jardin M, Janicot M, Page M, van Emelen K, Angibaud P. JNJ-26481585, a novel "second-generation" oral histone deacetylase inhibitor, shows broad-spectrum preclinical antitumoral activity. Clin Cancer Res. 2009;15(22):6841-6851