Vorinostat (SAHA, MK0683)

Catalog No.S1047

Vorinostat (suberoylanilide hydroxamic acid, SAHA) is an HDAC inhibitor with IC50 of ~10 nM in a cell-free assay.

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Vorinostat (SAHA, MK0683) Chemical Structure

Vorinostat (SAHA, MK0683) Chemical Structure
Molecular Weight: 264.3

Validation & Quality Control

Cited by 97 publications:

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Quality Control & MSDS

Related Compound Libraries

HDAC Inhibitors with Unique Features

Product Information

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  • Research Area
  • Vorinostat (SAHA, MK0683) Mechanism
  • Combination Therapy
    Combination Therapy

Product Description

Biological Activity

Description Vorinostat (suberoylanilide hydroxamic acid, SAHA) is an HDAC inhibitor with IC50 of ~10 nM in a cell-free assay.
Targets HDAC [1]
(Cell-free assay)
IC50 ~10 nM
In vitro Vorinostat inhibits the activities of HDAC1 and HDAC3 with IC50 of 10 nM and 20 nM, respectively. Vorinostat also results in a marked hyperacetylation of histone H4. [1] Vorinostat inhibits the growth of three prostate cancer cell lines LNCaP, PC-3 and TSU-Pr1 at micromolar concentrations (2.5-7.5 μM), and induces dose-dependent cell death in LNCaP cells. [2] Vorinostat treatment in MCF-7 cells inhibits cell proliferation at an IC50 of 0.75 μM resulting in the accumulation of cells in the G1 and G2-M phase of the cell cycle. Vorinostat also induces differentiation in the estrogen receptor-negative cell line SKBr-3 and the retinoblastoma-negative cell line MDA-468. [3] Vorinostat treatment at 1 μM for 8 hours or more is sufficient to irreversibly induce apoptosis of human multiple myeloma (MM) cells. The gene expression profiles of Vorinostat treated MM cells are not hallmarked by global transcriptional activation, but by coordinated transcriptional changes of specific functional groups of genes such as cytokine-induced proliferative/survival signaling cascades, oncogenes-tumor suppressor genes, regulators of apoptosis, DNA synthesis-repair and cell cycle, and proteasome-ubiquitin function. [4]
Cell Data
Cell LinesAssay TypeConcentrationIncubation TimeFormulationActivity DescriptionPMID
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VMRC-RCZMWnHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?M3fOd2lEPTB;M{[uN|AzOiEQvF2=NV3iZ|lGW0GQR1XS
NMC-G1MVHHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?NEGwZ|FKSzVyPUO3MlA2OjlizszNNXPnb|RvW0GQR1XS
D-336MGNIHyW3JIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=MXrJR|UxRTRzLki0OFIh|ryPNE\WWZNUSU6JRWK=
UACC-62NIqwOVBIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=Mn\xTWM2OD12Mj61OVA2KM7:TR?=NGfESmxUSU6JRWK=
C-33-ANGT3bGlIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=NEDxUJNKSzVyPUS1MlMzPTZizszNMkSxV2FPT0WU
HCC1937NX[zV2tTT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=NEfhdYFKSzVyPUS3MlE6PThizszNMX7TRW5ITVJ?
HDLM-2MlLuS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?MnSzTWM2OD12Nz6yNlg4KM7:TR?=MkX2V2FPT0WU
NCI-H727Moq2S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?MWTJR|UxRTR5LkK0OVEh|ryPMlPnV2FPT0WU
NYMmXXS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?NWPqc2VNUUN3ME20PU4{QTB6IN88US=>MkXrV2FPT0WU
NCI-H2291M2PZW2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7MoraTWM2OD12OT64N|AyKM7:TR?=MW\TRW5ITVJ?

... Click to View More Cell Line Experimental Data

In vivo Administration of Vorinostat (~100 mg/kg/day) significantly inhibits the growth of CWR22 human prostate xenografts in nude mice with tumor reductions of 78%, 97% and 97%, at doses of 25 mg/kg/day, 50 mg/kg/day and 100 mg/kg/day, respectively, compared with control. Vorinostat induces the accumulation of acetylated core histones and prostate-specific antigen mRNA expression in CWR22 cells, resulting in higher levels of serum prostate-specific antigen than predicted from tumor volume alone. [2] Oral administration of Vorinostat (0.67g/L) crosses the blood-brain barrier, increases histone acetylation in the brain, and dramatically improves the motor impairment in the R6/2 mice model of Huntington's disease. [5]
Features

Protocol(Only for Reference)

Kinase Assay:

[1]

Immunoprecipitation-HDAC assays The lysate of Jurkat cells is incubated for 1 hour on ice and cleared by centrifugation at 12,000 g for 10 minutes at 4 °C. Supernatants are precleared with 30 μL of 50% protein G-Sepharose slurry for 1 hour at 4 °C. Beads are pelleted by centrifugation and supernatants are incubated for 1 hour at 4 °C with 10 μg of IgG fraction from anti-HDAC1 or HDAC3 polyclonal antisera (preincubated 2 hours at room temperature with either the homologous or heterologous immunizing peptide). Both antisera are raised in rabbits against the carboxylterminal peptide of HDAC1 and HDAC3 by using synthetic peptides coupled to keyhole limpet hemocyanin. 30 μL of 50% protein G-Sepharose slurry is added for 1 hour at 4 °C. Immune complexes are pelleted by centrifugation and washed three times with 1 mL of lysis buffer. Beads are resuspended in 200 μL of HDAC buffer (20 mM Tris-HCl, pH 8.0/150 mM NaCl/10% glycerol), and the HDAC assay is performed with an 3H-acetylated peptide corresponding to amino acids 1-24 of histone H4. Released [3H]acetic acid is quantified by scintillation counting. For inhibitions studies, the immunoprecipitated complexes are preincubated with the different concentrations of Vorinostat for 30 minutes at 4 °C.

Cell Assay:

[2]

Cell lines LNCaP, PC-3, and TSU-Pr1
Concentrations Dissolved in DMSO, final concentrations ~7.5 μM
Incubation Time 1, 2, 3 and 4 days
Method

Cells are exposed to various concentrations of Vorinostat for 1, 2, 3 and 4 days. Cell viability is assessed by trypan blue dye exclusion.

Animal Study:

[2]

Animal Models Male BALB/c nude (nu/nu) mice implanted with CWR22 tumor cells
Formulation Dissolved in DMSO
Dosages ~100 mg/kg/day
Administration Injection i.p.

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

SpeciesMouseRatRabbitGuinea pigHamsterDog
Weight (kg)0.020.151.80.40.0810
Body Surface Area (m2)0.0070.0250.150.050.020.5
Km factor36128520
Animal A (mg/kg) = Animal B (mg/kg) multiplied by  Animal B Km
Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×  mouse Km(3)  = 11.2 mg/kg
rat Km(6)
1

References

[1] Richon VM, et al. Proc Natl Acad Sci U S A, 1998, 95(6), 3003-3007.

[2] Butler LM, et al. Cancer Res, 2000, 60(18), 5165-5170.

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Clinical Trial Information( data from http://clinicaltrials.gov, updated on 2016-06-25)

NCT Number Recruitment Conditions Sponsor
/Collaborators
Start Date Phases
NCT02589145 Recruiting Lymphoma M.D. Anderson Cancer Center|Celgene Corporation June 2016 Phase 1|Phase 2
NCT02707900 Not yet recruiting HIV-1 Infection Cynthia L Gay, MD|National Institute of Allergy and Infec  ...more Cynthia L Gay, MD|National Institute of Allergy and Infectious Diseases (NIAID)|University of North Carolina, Chapel Hill March 2016 Phase 1
NCT02638090 Recruiting Lung Cancer|Non-small Cell Lung Cancer H. Lee Moffitt Cancer Center and Research Institute|Merck  ...more H. Lee Moffitt Cancer Center and Research Institute|Merck Sharp & Dohme Corp. February 2016 Phase 1|Phase 2
NCT02619253 Recruiting Renal Cell Carcinoma|Urinary Bladder Neoplasms Roberto Pili|Indiana University January 2016 Phase 1|Phase 2
NCT02336074 Recruiting HIV Imperial College London|Medical Research Council|Universi  ...more Imperial College London|Medical Research Council|University of Oxford|University of Cambridge|Chelsea and Westminster NHS Foundation Trust|Royal Free Hospital NHS Foundation Trust|Brighton and Sussex University Hospitals NHS Trust|Guys and St Thomas NHS Foundation Trust|Central and North West London NHS Foundation Trust November 2015 Phase 2

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Chemical Information

Download Vorinostat (SAHA, MK0683) SDF
Molecular Weight (MW) 264.3
Formula

C14H20N2O3

CAS No. 149647-78-9
Storage 3 years -20℃powder
6 months-80℃in solvent
Synonyms N/A
Solubility (25°C) * In vitro DMSO 53 mg/mL (200.52 mM)
Ethanol 3 mg/mL (11.35 mM)
Water <1 mg/mL (<1 mM)
In vivo 2% DMSO+30% PEG 300+ddH2O 5mg/mL
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
Chemical Name N1-​hydroxy-​N8-​phenyl-octanediamide

Customer Product Validation(20)


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Source Nat Biotechnol 2011 29, 255-265. Vorinostat (SAHA, MK0683) purchased from Selleck
Method Immunofluorescence analysis
Cell Lines K562 cells
Concentrations 5 µM
Incubation Time 6 h
Results SAHA and other nonselective HDAC inhibitors increased steady-state acetylation of tubulin and histones manifested by the staining of acetylated microtubules and punctuate nuclear staining of acetylated histone.

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Source Nature 2011 471, 235-9. Vorinostat (SAHA, MK0683) purchased from Selleck
Method MTT cell viability assay
Cell Lines T-ALL cell lines
Concentrations 0.01-100 uM
Incubation Time 72 h
Results It examined responsiveness to dexamethasone and the class I/II HDAC inhibitor vorinostat in a panel of T-ALL cell lines with wild type or mutant CREBBP alleles. This demonstrated sensitivity to vorinostat at clinically useful concentrations (IC50 below 1礛) in the majority of cell lines tested.

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Source J Exp Med 2012 209, 35-50. Vorinostat (SAHA, MK0683) purchased from Selleck
Method Western blot
Cell Lines BM cells
Concentrations 100 ug/g
Incubation Time 11 days
Results Because of the observed decrease in global histone H3 acetylation, It treated NF-E2 tg mice with the HDAC-I vorinostat (SAHA) to restore physiological levels of histone H3 acetylation. Vorinostat treatment significantly increased H3 acetylation in NF-E2 tg animals.

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Source Clin Cancer Res 2012 18, 3822-33. Vorinostat (SAHA, MK0683) purchased from Selleck
Method Western blotting
Cell Lines LNCaP/22Rv1 cells
Concentrations 0.5/1μM
Incubation Time 24 h
Results The apoptosis induced by 0.5μM or 1μM SAHA was significantly increased in cells co-treated with 10μM bicalutamide; this was paralleled by demonstrable knockdown of c-FLIPL and c-FLIPS expression in these cells and by an enhanced level of cleaved PARP protein (Figure A/B). At the highest concentration of SAHA, co-treatment induced a rise of caspase-8 and caspase-3/7 activity to 245.3±28.9% and 265.5±15.6%, respectively (Figure C, left panel). Similarly, in LNCaP cells, caspase activity was augmented in response to SAHA and SAHA/bicalutamide; where there was a > 3-fold increase in caspase 3/7 activity with 2μM SAHA in combination with bicalutamide (Figure C, right panel). To confirm the importance of caspase-8 in initiating drug-induced apoptosis, caspase-8 activity was inhibited by the addition of Z-IETD-fmk prior to the drug treatments (Figure D).

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Source Diabetologia 2012 55, 2421-2431. Vorinostat (SAHA, MK0683) purchased from Selleck
Method Analyses of efficacy, potency and IC50
Cell Lines INS-1 cells
Concentrations
Incubation Time
Results HC toxin, CI-994, PCI24781 and LAQ824 all protected against cytokine-induced beta cell death, whereas the HDAC8-selective inhibitor PCI34051 showed no protection.

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Source Mol Carcinog 2012 52, 459-74. Vorinostat (SAHA, MK0683) purchased from Selleck
Method Real-Time Cell Growth Analysis
Cell Lines HT29/HCT116 cells
Concentrations 1-3 μM
Incubation Time 48 h
Results Treatment of HT29 and HCT116 CRC cells with increasing concentrations of butyrate or SAHA led to decreased proliferation over a 48 h period.

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Source J Neurochem 2012 123 Suppl 2, 108-15. Vorinostat (SAHA, MK0683) purchased from Selleck
Method Confocal microscopy/pixel-intensity measurements
Cell Lines mouse optic nerves
Concentrations 5 μM
Incubation Time
Results Suberoylanilide hydroxyamic acid (SAHA) preserves CFP (+) expression in mouse optic nerves (MONs) from older mice.

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Source J Transl Med 2011 9, 37. Vorinostat (SAHA, MK0683) purchased from Selleck
Method Tumor xenograft studies
Cell Lines female athymic nude mice
Concentrations 25 mg/kg
Incubation Time two weeks
Results SAHA , it self showed some anti-cancer effect in two tumor models ( Figure D and E). However, when mice were treated with S AHA followed by IL-13-PE, a significant decrease in tumor size was observed. In addition, 50% of mice showed complete elimination of their tumors in combination group. Next, we e valuated anti-cancer effect of combination of SAHA and IL-13-PE in IL-13Ra 2-positive pancreatic cancer model (HS766T and M IA -PaCa2). We observed that IL-13 - PE could significantly decrease tumor size in both IL-13Ra 2- positive tumors (Figure F and G). But when combined with SAHA , IL-13-PE not only decreased tumor size but also completely eliminated tumors in 66 to 83% of mice. These data suggest that SAHA can enhance anti-cancer effect of IL-13-PE even in IL-13Ra 2-positive pancreatic cancers.

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Source J Transl Med 2011 9, 37. Vorinostat (SAHA, MK0683) purchased from Selleck
Method qRT-PCR/immunostaining
Cell Lines female athymic nude mice
Concentrations 25 mg/kg
Incubation Time two weeks
Results Human IL-13Rα2 mRNA was significantly increased in tumors in both SAHA treated mice (Figure A). IL-13-PE treatment had no effect by itself but in combination with SAHA , a significant decrease in IL-13Rα2 expression was observed. In contrast, none of the organs except brain showed a modest increase in mouse IL-13Rα2 mRNA expression (Figure B). We also examined IL-13Rα2 protein expression by IHC. Similar to mRNA results, human IL-13Rα2 was dramatically increased in tumors from SAHA treated mice and when combined with IL-13 - PE, a decrease in IL-13Rα2 expression was observed ( Figure C ) . In normal tissues, mouse IL-13Rα2 was not detected or levels were below the detection limit of the assay in all organs examined (Figure D).

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Source Exp Dermatol 2010 19, 1096-1102. Vorinostat (SAHA, MK0683) purchased from Selleck
Method Tumor volume measurement
Cell Lines MyLa2059 cells
Concentrations 60 mg/kg
Incubation Time
Results The tumor growth was suppressed in mice treated with vorinostat when compared to mice treated with vehicle.

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Source Invest New Drugs 2012 30, 541-547. Vorinostat (SAHA, MK0683) purchased from Selleck
Method Western blot
Cell Lines BT-474 cells
Concentrations 0-10 μM
Incubation Time 8 h
Results Proteolytic processing of the caspase substrate PARP, a hall mark of apoptosis, increased in the BT-474 cells treated with SAHA and TRAIL.These results demonstrate that the combined treatment with SAHA and TRAIL induces significant apoptosis in these breast cancer cell lines.The activation of caspase-8 induced by TRAIL was clearly enhanced by pre-treatment with SAHA.The amount of intact Bid clearly diminished in the cells treated with both SAHA and TRAIL when compared to the cells treated with TRAIL or SAHA alone(Fig.a).The cellular levels of both cFLIPL and cFLIPS protein decreased in a dose-depen dent manner following exposure to SAHA, where as the levels of other prote ins in the TRAIL signalling pathway were not affected by SAHA (Fig.b)

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Source Invest New Drugs 2012 30, 541-547. Vorinostat (SAHA, MK0683) purchased from Selleck
Method Analysis of apoptosis
Cell Lines BT-474 cells, MDA-MB-231 cells
Concentrations 0-10 µM
Incubation Time 8 h
Results When the cells were exposed to SAHA for 8 h prior to the overni ght addition of TRAIL , apoptotic cell death was increased signifi cantly in both cell lines, particularly in the highly resistant BT-474 cells that over-expre s the ErbB2 /Her-2/ neu oncogene.

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Source Invest New Drugs 2012 30, 541-547. Vorinostat (SAHA, MK0683) purchased from Selleck
Method Western blot
Cell Lines BT-474 cells
Concentrations 5 µM
Incubation Time 8 h
Results Both MG132 and epoxomycin blocked the down-regulation of the two cFLIP isoforms induced by SAHA in BT-474 cells (Fig. a).even though siRNA treatment effectively depleted Itch/AIP4 in SAHA-treated cells, its depletion did not influence the down-regulation of cFLIP proteins (Fig. b).

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Source Invest New Drugs 2012 30, 541-547. Vorinostat (SAHA, MK0683) purchased from Selleck
Method Analysis of apoptosis, Western blot
Cell Lines BT-474 cells
Concentrations 0-10 μM
Incubation Time 8 h
Results Cells over-expressing cFLIPL were clearly more resistant to SAHA-induced sensitization to TRAIL-mediated apoptosis than cells expressing normal cFLIP levels (Fig. b)

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Source BMC Neurosci 2011 12, 50. Vorinostat (SAHA, MK0683) purchased from Selleck
Method Flow cytometry/Western blot
Cell Lines NSCs
Concentrations 1 μM
Incubation Time 48 h
Results Western blot analysis of whole cell extracts from adult NSCs treated with HDACi for 48 hours confirms SAHA and NaB treatment results in an increase inhistone3 (H3)acetylation levels when compared to DMSO and water vehicle controls (Figure a). Qu antita-tive an alysis of adult NSCs after 7-days HDACi treatment confirms SAHA and NaB treatment dramatically inhibits the formation of neurospheres (diameter≥ 50 μm) and increase the prevalence of small (≥ 4 cells,diameter < 50 μm) cell clusters in culture (Figure c ). flow cytometry reveals SAHA and NaB exposureresults in a 10 % reduction in cell viability compared to vehicle controls after 48 hours treatment (Figure d).

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Source BMC Neurosci 2011 12, 50. Vorinostat (SAHA, MK0683) purchased from Selleck
Method Flow cytometry/Fluorescence micrographs
Cell Lines NSCs
Concentrations 1 μM
Incubation Time 16 h
Results The addition of either SAHA or NaB significantly inhibited DNA synthesis in adult mouse NSCsin vitro. these flow cytometry data indicate that the suppression of NSC proliferation by SAHA and NaB result s in a G1-to-S phase block in cell cycle progression.

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Source BMC Neurosci 2011 12, 50. Vorinostat (SAHA, MK0683) purchased from Selleck
Method immunofluorescence cell fate assay/Western blot
Cell Lines NSCs
Concentrations 1 μM
Incubation Time 48 h
Results SAHA significantly reduced glial (p < 0.001) and oligodendrocyte (p < 0.05) differentiated cell fate in culture w hen compared to DMSO vehicle controls (Figure a). Fold-changes of b-catenin nuclear protein levels normalized to laminin A/C reveal marginal opposing +/- effects of SAHA (-1.49 ± 0 .11) and NaB (1.36 ± 0.33) treatment, indicating SAHA modulates cell fate via mechanisms independent of increased β-catenin stabilization.

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Source Pediatr Surg Int 2012 28, 579-89. Vorinostat (SAHA, MK0683) purchased from Selleck
Method DAPI staining and microscopy/ Western blot
Cell Lines HUH6 cells/ HepT1 cells
Concentrations 0.2/0.5 μM
Incubation Time 72 h
Results The effect of VX-680 and/or SAHA on cell morphology was similar in both, HepT1 and HUH6 cells (Fig. b). There was also a significant increase in both the nuclei and cell diameter in HepT1 cells when treated with VX–680 alone or in combination. In both HB cell. As shown in Fig. c, In both HB cell lines treatment with VX–680 and the combination with SAHA led to a decrease in histone H3 phosphorylation.

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Source Dr. Akihiko Yoshimura of Akihiko Yoshimura. Vorinostat (SAHA, MK0683) purchased from Selleck
Method Hoechst staining
Cell Lines C2C12 cells
Concentrations
Incubation Time
Results

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Source Dr. Zhang of Tianjin Medical University. Vorinostat (SAHA, MK0683) purchased from Selleck
Method Western blot
Cell Lines MDB-MA-231 cells
Concentrations 0-20 μM
Incubation Time
Results

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Tel: +1-832-582-8158 Ext:3

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