2. Immunology/Inflammation
  3. SOD
  4. ATN-224

ATN-224  (Synonyms: Bis(choline)tetrathiomolybdate)

Cat. No.: HY-16074 Purity: ≥98.0%
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ATN-224 is an oral Cu2+/Zn2+-superoxide dismutase 1 (SOD1) inhibitor. ATN-224 inhibits SOD1 activity in endothelial cells, an effect that is dose dependent with an IC50 of 17.5±3.7 nM.

For research use only. We do not sell to patients.

ATN-224 Chemical Structure

ATN-224 Chemical Structure

CAS No. : 649749-10-0

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Solid + Solvent
10 mM * 1 mL in DMSO
ready for reconstitution
 USD 88 In-stock
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10 mM * 1 mL in DMSO USD 88 In-stock
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10 mg USD 136 In-stock
50 mg USD 450 In-stock
100 mg USD 720 In-stock
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Customer Review

Based on 4 publication(s) in Google Scholar

ATN-224 Related Antibodies

Top Publications Citing Use of Products

    ATN-224 purchased from MedChemExpress. Usage Cited in: Anal Chem. 2018 Jan 16;90(2):1317-1324.  [Abstract]

    Illumination of the ・OH release along the deactivation of SOD1 by enzyme inhibitor ATN-224. The RAW 264.7 cells are treated by 100 μM ATN-224 for 0, 2, 6, 18 h, and then incubated with 10 μM P2 for 30 min. Scale bar: 10 μm. Fluorescence intensity (F) is displayed as yellow bars.

    • Biological Activity

    • Protocol

    • Purity & Documentation

    • References

    • Customer Review

    Description

    ATN-224 is an oral Cu2+/Zn2+-superoxide dismutase 1 (SOD1) inhibitor. ATN-224 inhibits SOD1 activity in endothelial cells, an effect that is dose dependent with an IC50 of 17.5±3.7 nM.

    IC50 & Target

    IC50: 17.5±3.7 nM (SOD1, in endothelial cells)[1]
    In Vitro

    ATN-224 has a specific and high affinity for copper ions (108 mol/L-1) and shows no binding to calcium, iron, magnesium, zinc, or manganese ions at concentrations up to 1 mM as determined by isothermal titration calorimetry. ATN-224 inhibits the proliferation of both HUVEC (IC50=1.4±0.3 μM; n=5). ATN-224 is also able to inhibit the activity of purified bovine SOD1 with an IC50 of 0.33±0.03 μM after 24 hours of incubation. The SOD1 inhibition by ATN-224 is time dependent, reaching maximal inhibition at ~16 hours. ATN-224 seems to inhibit SOD1 by depleting the enzyme of copper. ATN-224 is able to inhibit SOD1 activity in endothelial cells, an effect that is dose dependent with an IC50 of 17.5±3.7 nM. ATN-224 inhibits FGF-2-induced ERK1/2 phosphorylation in a dose-dependent and time-dependent manner with an IC50 between 1.25 and 2.5 μM, consistent with the IC50 for the inhibition of proliferation[1]. ATN-224 is an orally-available inorganic small molecule that inhibits the copper/zinc-dependent enzyme, superoxide dismutase 1 (Cu/Zn-SOD1), in endothelial and tumor cells[2].

    MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

    ATN-224 Related Antibodies
    In Vivo

    ATN-224 also significantly (P<0.05) inhibits angiogenesis in the Matrigel plug model in mice either when added directly to the plug or when given by oral gavage. Inhibition of angiogenesis when ATN-224 is given by oral gavage occurred before there is measurable depletion of copper in either plasma or copper from the Matrigel plug. This result shows that ATN-224 inhibits angiogenesis independently of copper depletion[1].

    MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
    Molecular Weight

    432.56

    Formula

    C10H28MoN2O2S4
    CAS No.
    Appearance

    Solid

    Color

    Pink to red

    SMILES

    OCC[N+](C)(C)C.OCC[N+](C)(C)C.[S-][Mo]([S-])(=S)=S
    Shipping

    Room temperature in continental US; may vary elsewhere.
    Storage
    Powder -20°C 3 years
    4°C 2 years
    In solvent -80°C 6 months
    -20°C 1 month
    Solvent & Solubility
    In Vitro: 

    DMSO : 50 mg/mL (115.59 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 2.3118 mL 11.5591 mL 23.1182 mL
    5 mM 0.4624 mL 2.3118 mL 4.6236 mL
    View the Complete Stock Solution Preparation Table

    * Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
    Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

    • Molarity Calculator

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    Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

    Mass
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    Concentration
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    Volume
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    Molecular Weight *

    Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

    This equation is commonly abbreviated as: C1V1 = C2V2

    Concentration (start)

    C1

    ×
    Volume (start)

    V1

    =
    Concentration (final)

    C2

    ×
    Volume (final)

    V2

    In Vivo:

    Select the appropriate dissolution method based on your experimental animal and administration route.

    For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
    To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
    The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

    • Protocol 1

      Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

      Solubility: ≥ 2.5 mg/mL (5.78 mM); Clear solution

      This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

      Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

      Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
    • Protocol 2

      Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

      Solubility: ≥ 2.5 mg/mL (5.78 mM); Clear solution

      This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

      Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

      Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
    In Vivo Dissolution Calculator
    Please enter the basic information of animal experiments:

    Dosage

    mg/kg

    Animal weight
    (per animal)

    g

    Dosing volume
    (per animal)

    μL

    Number of animals

    Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
    Please enter your animal formula composition:
    %
    DMSO +
    % 
    +
    %
    Tween-80 +
    %
    Saline
    Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
    The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
    Calculation results:
    Working solution concentration: mg/mL
    Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).
    The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
    Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
     If the continuous dosing period exceeds half a month, please choose this protocol carefully.
    Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
    Purity & Documentation

    Purity: ≥98.0%

    COA (247 KB) HNMR (165 KB)

    Handling Instructions (2659 KB)
    References
    Kinase Assay
    [1]

    Cells are plated in a six-well format (100-300,000 per well) and incubated with ATN-224 at indicated concentrations and for indicated times. Cells are then stimulated with 10 ng/mL FGF-2 for various times and lysed. Lysates are subjected to Western blot analysis using an antibody specific for phosphorylated p44/42 mitogen-activated protein kinase (Thr202/Tyr204) with appropriate signal correction using an antibody specific for p44/42 mitogen-activated protein kinase[1].

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.
    Cell Assay
    [1]

    Human umbilical vein endothelial cells (HUVEC) are maintained in M200/LSGS medium and cells are used between passages 2 and 4 for all experiments. For proliferation assay, cells are plated at 3,000 per well on 0.1% gelatin in M200/2% FBS for 4 hours and then stimulated with 2 ng/mL FGF-2 in the presence or absence of ATN-224 up to 48 hours. HUVEC proliferation is determined using either the Alamar Blue or the MTT assay. Multiple myeloma MM1S cells are grown in RPMI 1640 with 10% fetal bovine serum and 2 mM L-glutamine. HL-60 promyelocytic leukemia cells and MOLT-4 acute myeloblastic leukemia cells are plated at 400,000/mL in T-75 flasks and incubated for 48 to 96 hours for proliferation assays. MMS1 cell proliferation is determined using calcein AM[1].

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.
    Animal Administration
    [1]

    Mice[1]
    Cold Matrigel (500 μL) is mixed with 800 ng/mL FGF-2 or 300 ng/mL VEGF and heparin (50 μg/mL). Negative control plugs did not contain the proangiogenic factors. The Matrigel mixture is injected s.c. into 4- to 8-week-old female BALB/c nude mice. In some experiments, either ATN-224 (94 μM) with or without Mn-TBAP (100 μM) or water is added directly to the Matrigel plug in the treated and negative control groups, respectively. Alternatively, mice are treated by oral gavage either with distilled water or ATN-224 everyday from Monday to Friday. Animals are sacrificed and the plugs are recovered 5 days after plug injection. The plugs are then minced and homogenized with a tissue homogenizer, and hemoglobin levels in the plugs are determined using Drabkin's solution.

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.
    References

    Complete Stock Solution Preparation Table

    * Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
    Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

    Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
    DMSO 1 mM 2.3118 mL 11.5591 mL 23.1182 mL 57.7955 mL
    5 mM 0.4624 mL 2.3118 mL 4.6236 mL 11.5591 mL
    10 mM 0.2312 mL 1.1559 mL 2.3118 mL 5.7795 mL
    15 mM 0.1541 mL 0.7706 mL 1.5412 mL 3.8530 mL
    20 mM 0.1156 mL 0.5780 mL 1.1559 mL 2.8898 mL
    25 mM 0.0925 mL 0.4624 mL 0.9247 mL 2.3118 mL
    30 mM 0.0771 mL 0.3853 mL 0.7706 mL 1.9265 mL
    40 mM 0.0578 mL 0.2890 mL 0.5780 mL 1.4449 mL
    50 mM 0.0462 mL 0.2312 mL 0.4624 mL 1.1559 mL
    60 mM 0.0385 mL 0.1927 mL 0.3853 mL 0.9633 mL
    80 mM 0.0289 mL 0.1445 mL 0.2890 mL 0.7224 mL
    100 mM 0.0231 mL 0.1156 mL 0.2312 mL 0.5780 mL
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    ATN-224 Related Classifications

    Help & FAQs
    • Do most proteins show cross-species activity?

      Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

    Keywords:

    ATN-224649749-10-0Bis(choline)tetrathiomolybdateATN224ATN 224SODSuperoxide DismutaseInhibitorinhibitorinhibit

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