Decitabine

Decitabine is a potent inhibitor of DNA methylation, used to treat myelodysplastic syndrome (MDS).

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Decitabine Chemical Structure

Decitabine Chemical Structure
Molecular Weight: 228.21

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Product Information

  • Compare DNA Methyltransferase Inhibitors
    Compare DNA Methyltransferase Products
  • Research Area
  • Combination Therapy
    Combination Therapy

Product Description

Biological Activity

Description Decitabine is a potent inhibitor of DNA methylation, used to treat myelodysplastic syndrome (MDS).
Targets DNA methylation [1]
(HL-60, KG1a cells)
In vitro Decitabine inhibits cell growth in a dose and time-dependent manner with IC50 of approximately 438 nM and 43.8 nM for 72 hours and 96 hours exposure in HL-60 and KG1a leukemic cells, respectively. [1] A recent study shows that Decitabine exhibits high anti-proliferative and pro-apoptotic activity against anaplastic large cell lymphoma (ALCL), and inhibits [3H]–thymidine uptake in KARPAS-299 cells with EC50 of 0.49 μM. [2]
Cell Data
Cell LinesAssay TypeConcentrationIncubation TimeFormulationActivity DescriptionPMID
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... Click to View More Cell Line Experimental Data

In vivo In a ALK+ KARPAS-299 murine xenograft model, Decitabine at a dose of 2.5 mg/kg causes increased apoptosis and reduced proliferation of tumor cells, and also results in demethylation of tumor suppressor p16INK4A. [2]
Features

Protocol(Only for Reference)

Kinase Assay: [1]

DNA synthesis assay The rate of DNA synthesis is measured by the incorporation of radioactive thymidine into DNA. HL-60 and KG1a cells are suspended in 2 mL RPMI medium containing 10% fetal serum in 6-well (35 mm diameter) dishes and incubated with different concentrations of corresponding drugs for 48 hours (drugs are added simultaneously). At 48 hours, 0.5 μCi [3H] thymidine (6.7 Ci/mmol) is added to each well and incubated for an additional 24 hours. The cells are placed on GF/C glass fiber filters (2.4 cm diameter), washed with cold 0.9% NaCl, 5% cold trichloroacetic acid and ethanol. The filters containing the DNA are then dried, placed in EcoLite scintillation liquid (ICN) and the radioactivity measured using Beckman LS 6000IC scintillation counter. The IC50 is defined as the concentration of drug that inhibits 50% of the DNA synthesis of the leukemic cell lines from the dose–response curve.

Cell Assay: [1]

Cell lines HL-60 and KG1a
Concentrations 0-500 nM
Incubation Time 96 hours
Method For the growth inhibition assay, cells in log phase are placed in 5 mL of medium. Different concentrations of Decitabine are added to the medium simultaneously. Cell counts are performed at the indicated times using a model ZM Coulter Counter. The concentration that produces 50% inhibition of growth (IC50) is determined from the growth curves of the drug treated leukemic cell lines.

Animal Study: [2]

Animal Models KARPAS-299 human cells are inoculated subcutaneously into the right and left flanks of the mice.
Formulation Decitabine is dissolved in sterile PBS .
Dosages ≤2.5 mg/kg
Administration Administered via 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] Shaker S, et al. Leuk Res, 2003, 27(5), 437-444.

[2] Hassler MR, et al. Biochimie, 2012, doi.org/10.1016/j.biochi.2012.05.029.

Clinical Trial Information( data from http://clinicaltrials.gov, updated on 2016-01-10)

NCT Number Recruitment Conditions Sponsor
/Collaborators
Start Date Phases
NCT02634827 Recruiting Secondary Acute Myeloid Leukemia|Untreated Adult Acute Myeloid Leukemia Mayo Clinic|National Cancer Institute (NCI) January 2016 Phase 2
NCT02632721 Not yet recruiting Leukemia, Myeloid, Acute Boehringer Ingelheim January 2016 Phase 2
NCT02564536 Not yet recruiting Chronic Myelomonocytic Leukemia|Juvenile Myelomonocytic Leukemia|Atypical Chronic Myeloid Leukemia|Myeloproliferative Neoplasm|Myelodysplastic Synd  ...more Chronic Myelomonocytic Leukemia|Juvenile Myelomonocytic Leukemia|Atypical Chronic Myeloid Leukemia|Myeloproliferative Neoplasm|Myelodysplastic Syndromes|Myelofibrosis Washington University School of Medicine|CTI BioPharma January 2016 Phase 0
NCT02323607 Not yet recruiting Recurrent Adult Acute Myeloid Leukemia|Secondary Acute Myeloid Leukemia|Therapy-Related Acute Myeloid Leukemia|Untreated Adult Acute Myeloid Leukemia Bhavana Bhatnagar|CTI BioPharma|Ohio State University Com  ...more Bhavana Bhatnagar|CTI BioPharma|Ohio State University Comprehensive Cancer Center December 2015 Phase 1
NCT02469415 Recruiting Leukemia M.D. Anderson Cancer Center|CTI BioPharma September 2015 Phase 2

view more

Chemical Information

Download Decitabine SDF
Molecular Weight (MW) 228.21
Formula

C8H12N4O4

CAS No. 2353-33-5
Storage 3 years -20℃powder
6 months-80℃in solvent
Synonyms Deoxycytidine
Solubility (25°C) * In vitro DMSO 45 mg/mL (197.18 mM)
Water 10 mg/mL (43.81 mM)
Ethanol <1 mg/mL (<1 mM)
In vivo 30% propylene glycol, 5% Tween 80, 65% D5W 30 mg/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 4-amino-1-((2R,4S,5R)-4-hydroxy-5-(hydroxymethyl)-tetrahydrofuran-2-yl)-1,3,5-triazin-2(1H)-one

Customer Product Validation (3)


Click to enlarge
Rating
Source PLoS One 2014 9(5), e97719. Decitabine purchased from Selleck
Method Western blot
Cell Lines SW1116, LOVO cells
Concentrations 10 uM
Incubation Time 48 h
Results Decitabine (10 uM) could not lead to significant alterations in AKT, or mTOR activity, as assessed by Western blot for phosphorylation of AKT, mTOR and S6K. Additionally, it observed minimal reductions of phosphorylation of AKT, mTOR and S6K with 5 uM gefitinib in SW1116 and 4 礛 in LOVO cells. However, the combination of decitabine and gefitinib completely abrogated AKT and mTOR activities in SW1116 and LOVO cells.

Click to enlarge
Rating
Source PLoS One 2014 9(5), e97719. Decitabine purchased from Selleck
Method Annexin V-FITC/PI analysis
Cell Lines SW1116, LOVO cells
Concentrations 10 uM
Incubation Time 48 h
Results SW1116 and LOVO cells were treated with the two compounds, alone or in combination, for 48 h and then cell apoptosis was determined by Annexin V-FITC and propidium iodide (PI) staining and flow cytometry analysis. Treatment with gefitinib (2 uM) or decitabine (10 uM) alone had weak effects on apoptosis in SW1116 cells. However, there was a significantly higher apoptosis rate found upon treatment with the combination of gefitinib and decitabine.

Click to enlarge
Rating
Source Leuk Res 2014 38(3), 402-10. Decitabine purchased from Selleck
Method Bisulfite sequencing analysis
Cell Lines NB4 cells
Concentrations 10 uM
Incubation Time 24 h
Results Mature miR-125a expression increased with increasing amounts of decitabine treatment after 24 hours.

Product Use Citation (6)

Tech Support & FAQs

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.

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