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Daminozide

Alias: DIMG; B 995; Aminozide
Cat No.:V19151 Purity: ≥98%
Daminozide is a plant growth regulator and a selective inhibitor of human KDM2/7 histone demethylase, with IC50s of 0.55, 1.5 and 2.1 μM for PHF8, KDM2A and KIAA1718 respectively.
Daminozide
Daminozide Chemical Structure CAS No.: 1596-84-5
Product category: New1
This product is for research use only, not for human use. We do not sell to patients.
Size Price Stock Qty
1g
5g
Other Sizes
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Product Description
Daminozide is a plant growth regulator and a selective inhibitor of human KDM2/7 histone demethylase, with IC50s of 0.55, 1.5 and 2.1 μM for PHF8, KDM2A and KIAA1718 respectively. Daminozide is >100-fold selective for the KDM2/7 subfamily compared to other demethylase subfamily members tested.
Biological Activity I Assay Protocols (From Reference)
ln Vitro
Compared to other demethylase subfamily members examined (IC50=127 μM for KDM3A, or higher for other demethylases), dacazine is >100-fold more selective for the KDM2/7 subfamily [1].
ln Vivo
The model organism Drosophila melanogaster is teratogenic and genotoxic when exposed to doses of 200–20,000 ppm of dinonozide [2].
ADME/Pharmacokinetics
Absorption, Distribution and Excretion
This study selected miniature pigs as experimental animals because their stomach conditions are similar to those of humans. The experimental animals were orally administered a radiolabeled test substance at a dose of 5 mg/kg body weight. The results showed that the substance was rapidly excreted, with approximately 60% eliminated within 48 hours. The highest concentration of the metabolite (UDMH) in urine was 2.8 ppm; the concentration in feces was lower. Furthermore, a radiolabeled 1,1-dimethylnitrosamine at a concentration of 0.69 ppm was detected in the urine of the experimental animals. A second study used radiolabeled daminozide for topical application to the skin at concentrations of 0.5, 0.05, or 0.005 mg/cm² for up to 24 hours. Within 24 hours, the absorbed and excreted dose in the experimental animals was as high as 1.8%. The distribution and metabolic pathways of (14)C daminozide in seedlings of Acer buergerianum and Platanus orientalis were studied using autoradiography. Radioactivity was detected in all parts of the plants. (14)C is concentrated in the meristematic regions of leaves. Analysis of extracts from red oak (Quercus rubra) treated with (14)C-labeled hydrazine maleate or daminozide showed that both growth inhibitors distributed towards the apex and base within 22 days. When leaves of 3-year-old apple trees (Cox's Orange pippin variety) were smeared with (14)C-labeled daminozide, more than half of the radioactive material accumulated in the fruit. Metabolites in the plant include 1,1-dimethylhydrazine.
Toxicity/Toxicokinetics
Toxicity Data
LC50 (Rat) > 147,000 mg/m³/1h
5,000 mg/kg LD50 (Mouse, oral): 6,300 mg/kg LD50 (Mouse, intraperitoneal injection): 1,325 mg/kg LC50 (Rat, inhalation): > 20 mg/L/hr
References

[1]. Rose NR, Woon EC, Tumber A, Plant growth regulator daminozide is a selective inhibitor of human KDM2/7 histone demethylases. J Med Chem. 2012 Jul 26;55(14):6639-43.

[2]. Exploration of teratogenic and genotoxic effects of fruit ripening retardant Alar (Daminozide) on model organism Drosophila melanogaster. Interdiscip Toxicol. 2018 May;11(1):27-37.

Additional Infomation
According to the U.S. Environmental Protection Agency (EPA), daminohydrazide may be carcinogenic. 2,2-Dimethylhydrazine succinate is a tasteless white crystal or powder. (NTP, 1992) Daminohydrazide is a straight-chain fatty acid. Daminohydrazide—also known as Alar, Kylar, B-NINE, DMASA, SADH, or B 995—is a plant growth regulator, a chemical sprayed on fruit to regulate its growth, facilitate harvesting, and prevent apples from falling from the tree before ripening. This ensures that apples remain bright red and firm during storage. Alar was first approved for use in the U.S. in 1963, primarily for apples, until 1989, when manufacturers voluntarily withdrew the product after the EPA proposed banning it due to its high carcinogenic risk to consumers.
These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C6H12N2O3
Molecular Weight
160.173
Exact Mass
160.084
CAS #
1596-84-5
PubChem CID
15331
Appearance
White to off-white solid powder
Density
1.2±0.1 g/cm3
Boiling Point
329.5±44.0 °C at 760 mmHg
Melting Point
162-164 °C(lit.)
Flash Point
153.1±28.4 °C
Vapour Pressure
0.0±1.6 mmHg at 25°C
Index of Refraction
1.497
LogP
-0.5
Hydrogen Bond Donor Count
2
Hydrogen Bond Acceptor Count
4
Rotatable Bond Count
4
Heavy Atom Count
11
Complexity
156
Defined Atom Stereocenter Count
0
InChi Key
NOQGZXFMHARMLW-UHFFFAOYSA-N
InChi Code
InChI=1S/C6H12N2O3/c1-8(2)7-5(9)3-4-6(10)11/h3-4H2,1-2H3,(H,7,9)(H,10,11)
Chemical Name
4-(2,2-dimethylhydrazinyl)-4-oxobutanoic acid
Synonyms
DIMG; B 995; Aminozide
HS Tariff Code
2934.99.9001
Storage

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Shipping Condition
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
Solubility Data
Solubility (In Vitro)
DMSO : ~106.67 mg/mL (~665.98 mM)
H2O : ~100 mg/mL (~624.34 mM)
Solubility (In Vivo)
Solubility in Formulation 1: ≥ 2.5 mg/mL (15.61 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

Solubility in Formulation 2: ≥ 2.5 mg/mL (15.61 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C,1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (15.61 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.


Solubility in Formulation 4: 100 mg/mL (624.34 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication (<60°C).

 (Please use freshly prepared in vivo formulations for optimal results.)
Preparing Stock Solutions 1 mg 5 mg 10 mg
1 mM 6.2434 mL 31.2168 mL 62.4337 mL
5 mM 1.2487 mL 6.2434 mL 12.4867 mL
10 mM 0.6243 mL 3.1217 mL 6.2434 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.

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What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?
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What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:
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Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
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Definitions of molecular mass, molecular weight, molar mass and molar weight:
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In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)
Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)
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Calculation results

Working concentration mg/mL;

Method for preparing DMSO stock solution mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.

Method for preparing in vivo formulation:Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.

(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
             (2) Be sure to add the solvent(s) in order.

Biological Data
  • Heatmap of JmjC demethylase inhibition by a set of 2OG analogues Daminozide 19 is selective for KDM2A. Tricarboxylic acid cycle intermediates succinate 1 and fumarate 2 were generally poor demethylase inhibitors, though KDM4E7 was an exception to this trend. (R)- and (S)-2-hydroxyglutarate enantiomers, produced by gain-of-function mutations to isocitrate dehydrogenase were inhibitors of the KDM2, KDM3 and KDM4 histone demethylases.12 The catechols 16 and 17, bipyridyl 18 and 8-hydroxyquinoline 21 inhibited all demethylases screened. By contrast, 5 most potently inhibited PHD2 (prolyl hydroxylase domain enzyme isoform 2). Each compound was screened in an AlphaScreen assay and is represented as % inhibition of the enzyme at 20 μM compound. J Med Chem . 2012 Jul 26;55(14):6639-43.
  • Crystal structures reveal the mode of inhibition of the KDM2/7 subfamily by daminozide PHF8 (with Zn(II) substituting for Fe(II),) and KDM4A (with Ni(II) substituting for Fe(II)) and FIH (with Zn(II) substituting for Fe(II)) (Table S3). Daminozide binds the metal through its hydrazide amine lone pair and carbonyl oxygen. Two distinct orientations of daminozide are observed in the two KDM4A molecules in the asymmetric unit; both are shown (see Table S2 for electron density maps). Selectivity of daminozide for the KDM2/7 subfamily may arise because they possess a hydrophobic region (Tyr257, Val255 and Ile313) into which the two daminozide methyl groups may bind. In contrast, the equivalent regions of KDM4A, FIH and the other tested demethylases/oxygenases are more hydrophilic. J Med Chem . 2012 Jul 26;55(14):6639-43.
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