Quisinostat (JNJ-26481585)

 Distributor Tel: +886-2-2827-1197jimmy@new.abscience.com.tw

Biological Activity

Description	JNJ-26481585 is a novel second-generation HDAC inhibitor with highest potency for HDAC1 with IC50 of 0.11 nM, modest potent to HDACs 2, 4, 10, and 11; greater than 30-fold selectivity against HDACs 3, 5, 8, and 9 and lowest potency to HDACs 6 and 7. Phase 2.
Targets	HDAC1	HDAC2	HDAC4	HDAC10	HDAC11
IC50	0.11 nM	0.33 nM	0.64 nM	0.46 nM	0.37 nM [1]
In vitro	JNJ-26481585 exhibits broad spectrum antiproliferative activity in solid and hematologic cancer cell lines, such as all lung, breast, colon, prostate, brain, and ovarian tumor cell lines, with IC50 ranging from 3.1-246 nM, which is more potent than vorinostat, R306465, panobinostat, CRA-24781, or mocetinostat in various human cancer cell lines tested. [1] A recent study shows that JNJ-26481585 promotes myeloma cell death at low nanomolar concentrations by resulting in Mcl-1 depletion and Hsp72 induction. [2]
In vivo	In an HDAC1-responsive A2780 ovarian tumor screening model, JNJ-26481585 dosing at its maximal tolerated dose (10 mg/kg i.p. and 40 mg/kg p.o.) for 3 days leads to an HDAC1-regulated fluorescence , which predicts tumor growth inhibition. Furthermore, JNJ-26481585 also shows more potent inhibitory effects on the growth of C170HM2 colorectal liver metastases than 5-fluorouracil/Leucovorin. [1]
Features	An orally bioavailable, second-generation, hydroxamic acid-based HDAC inhibitor.

Protocol(Only for Reference)

Kinase Assay: ^[1]

HDAC activity assays

In all cases, full-length HDAC proteins are expressed using baculovirus-infected Sf9 cells. In addition, HDAC3 is coexpressed as a complex with human NCOR2. For assessing activity of HDAC1-containing cellular complexes, immunoprecipitated HDAC1 complexes are incubated with an [3H]acetyl- labeled fragment of histone H4 peptide [biotin-(6-aminohexanoic)Gly-Ala-(acetyl[3H])Lys-Arg-His-Arg-Lys-Val-NH2] in a total volume of 50μL enzyme assay buffer (25mM HEPES (pH 7.4), 1 M sucrose, 0.1 mg/mL BSA and 0.01% (v/v) Triton X-100). Incubation is performed for 45 minutes at 37 °C (immunoprecipitates) or 30 min at room temperature. Before addition of substrate, HDAC inhibitors are added at increasing concentrations and preincubated for 10 minutes at room temperature. After incubation, the reaction is quenched with 35μL stop buffer (1 M HCl and 0.4 M acetic acid). Released [3H]acetic acid is extracted with 800μL ethyl acetate and quantified by scintillation counting. Equal amounts of HDAC1 are immunoprecipitated as indicated by Western blot analysis. HDAC1 activity results are presented as mean ± SD of three independent experiments on a single lysate.

Cell Assay: Cell proliferation assays

Cell lines	NCL-H2106, Colo699 and LNCAP cells
Concentrations	0-300 nM
Incubation Time	24 hours
Method	All cell lines are obtained from American Type Culture Collection and cultured according to instructions. The effect of HDAC inhibitors on cell proliferation is measured using an MTT. Proliferation of non–small cell lung carcinoma (NSCLC) cell lines is assessed using an Alamar Blue–based assay. For proliferation of hematologic cell lines, cells are incubated for 72 hours and the cytotoxic activity is evaluated by MTS assay. Data are presented as mean IC50 or IC40 ± SD of at least three independent experiments.

Animal Study: ^[1]

Animal Models	HCT116 human colon carcinoma cells are injected s.c. into the inguinal region of athymic male NMRI nu/nu mice, C170HM2 cell suspensions are injected into the peritoneal cavity of male MFI nude mice.
Dosages	≤10 mg/kg
Administration	Administered via both p.o. and i.p.
Solubility	15% Captisol, 30 mg/mL

1

References

Clinical Trial Information( data from http://clinicaltrials.gov)

NCT Number	Recruitment	Conditions	Sponsor /Collaborators	Start Date	Phases
NCT00677105	Completed	Lymphoma|Neoplasms	Johnson & Johnson Pharmaceutical Research & Development, L.L.C.|Janssen Pharmaceutica N.V., Belgium	2007-08	Phase 1
NCT00676728	Terminated	Advanced or Refractory Leukemia|Myelodysplastic Syndromes	Johnson & Johnson Pharmaceutical Research & Development, L.L.C.	2008-12	Phase 1
NCT01464112	Recruiting	Multiple Myeloma	Janssen Research & Development, LLC	2011-09	Phase 1
NCT01486277	Active, not recruiting	Cutaneous T-cell Lymphoma; Histone Deacetylase Inhibitor; Lymphoma; Oncology	Janssen Research & Development, LLC	2011-11	Phase 2

Chemical Information

Download Quisinostat (JNJ-26481585) SDF

Chemical Name	N-hydroxy-2-(4-(((1-methyl-1H-indol-3-yl)methylamino)methyl)piperidin-1-yl)pyrimidine-5-carboxamide

Preparing Stock Solutions

Stock Solution (1ml DMSO)	1mM	10mM	20mM	30mM
Mass(mg)	0.394476	3.94476	7.88952	11.83428

Customer Reviews (1)

Click to enlarge	Rating
	Source	Dr. Zhang of Tianjin Medical University. Quisinostat (JNJ-26481585) purchased from Selleck
	Method	Western blot
	Cell Lines	MDA-MB-231 cells
	Concentrations	0-20 μM
	Incubation Time
	Results	Increased Histone H3 acetylation in MDA-MB-231 cells treated with JNJ-26481585 was observed.

Product Citations (1)

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  Selective cancer targeting with prodrugs activated by histone deacetylases and a tumour-associated protease. [Ueki N, et al. Nat Commun 2013;4:2735]