Pyroxamide (NSC 696085)

Catalog No.S2190 Batch:S219001

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Technical Data

Formula

C13H19N3O3
Molecular Weight 265.31 CAS No. 382180-17-8
Solubility (25°C)* In vitro DMSO 17 mg/mL (64.07 mM)
Ethanol 2 mg/mL (7.53 mM)
Water Insoluble
In vivo (Add solvents to the product individually and in order)
Clear solution
5%DMSO 40%PEG300 5%Tween80 50%ddH2O
0.85mg/ml Taking the 1 mL working solution as an example, add 50 μL of 17 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results. 
Clear solution
5% DMSO 95% Corn oil
0.85mg/ml Taking the 1 mL working solution as an example, add 50 μL of 17 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results. 
* <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.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description Pyroxamide(NSC 696085) is a potent inhibitor of affinity-purified HDAC1 with ID50 of 100 nM. It also induces growth suppression and cell death in human rhabdomyosarcoma in vitro.
In vitro

Pyroxamide causes the accumulation of acetylated core histones in MEL cells cultured with the agent. Pyroxamide, at micromolar concentrations, induces terminal differentiation and inhibits proliferation of murine erythroleukemia(MEL) cells. Pyroxamide (1.25-20 mM) causes dose-dependent inhibition of growth by cell cycle arrest of prostate carcinoma (LNCaP), neuroblastoma (KCN-69n), and bladder carcinoma (T24) cells in culture, with similar efficacy in all of the cell lines. [1] A concentration of 1.25-20.0 μM Pyroxamide causes a dose-dependent decrease in viable cell number and an increase in percentage of dead cells over time in two Rhabdomyosarcoma cell lines, RD (embryonal ) and RH30B (alveolar). Accumulation of acetylated histones and induction of p21/WAF1 expression are obersevd in cells exposed to Pyroxamide. [2] Pyroxamide shows a dose- and time-dependent proliferation inhibition, induction of apoptosis and histone H4 hyperacetylation in three B-cell precursor (BCP)-acute lymphoblastic leukemia (ALL) cell lines (Reh, Nalm6, Z33). The calculated IC50 after 96 hours of Pyroxamide incubation are 2-6 μM. [3]
In vivo

Administration of 100 or 200 mg/kg of Pyroxamide daily for 21 days causes significant, dose-dependent suppression of the growth of the tumor xenograft. A dose of 300 mg/kg Pyroxamide is lethal to all of the mice in the treatment group within 1 week. After in vivo administration of Pyroxamide, accumulation of acetylated histones and a dose-dependent increase in the expression of p21/WAF1 protein level are observed. [1]

Protocol (from reference)

Kinase Assay:

[1]
  • HDAC Inhibition Assays

    A MEL cell line expressing the epitope Flag-tagged HDAC1 is generated. HDAC1-Flag is affinity purified by immune-precipitation using M2 anti-Flag antibody-coated agarose, followed by elution from the agarose using the Flag peptide. [3H]acetate-labeled cellular histones are prepared from MEL cells and are used as a substrate for the HDAC activity assay. Released [3H] acetic acid is quantified by scintillation counting. For inhibition studies, the enzyme preparations are preincubated with Pyroxamide (10 to 100,000 nM) for 30 minutes at 4 °
Cell Assay:

[1]
  • Cell lines

    MEL Ds19/Sc9 cells

  • Concentrations

    0.625-8 μM

  • Incubation Time

    5 days

  • Method

    1-2 × 105 cells/mL MEL Ds19/Sc9 cells are cultured with increasing concentrations of Pyroxamide (0.625-8 μM) for 5 days, at which time the Benzidine positivity is determined as an indicator of terminal differentiation. MEL cell density is measured using a Coulter counter and viability is assessed by trypan blue exclusion.
Animal Study:

[1]
  • Animal Models

    Human CWR22 prostate cancer xenograft is s.c. injected in nude mice.

  • Dosages

    100, 200, or 300 mg/kg

  • Administration

    Administered daily via i.p. injection.

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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