| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg | |||
| Other Sizes |
| Targets |
p70S6 kinase（IC50 = 2.8 μM in kinase activity assay; inhibits Thr389 phosphorylation）[1]
Proteasome（chymotrypsin-like activity, IC50 = 5.6 μM）[1] |
|---|---|
| ln Vitro |
In U937 cells, DD1 (20 μM; 96 h) exhibits dose- and time-dependent antiproliferative action [1]. In the G2/M phase, DD1 causes cell cycle arrest, which results in apoptosis [1]. Bax overexpression, Bad dephosphorylation, and mitochondrial membrane depolarization are all brought on by DD1 (10, 20 μM; 48 h) [1]. In U937 lysates, DD1 (2–20 μM; 40 hours) shows inhibitory effects on chymotrypsin-like activity [1].
Anti-proliferative activity in myeloid tumor cells: DD1 inhibitor (1–20 μM) dose-dependently inhibits proliferation of U937 (monocytic leukemia), HL-60 (promyelocytic leukemia), and K562 (chronic myeloid leukemia) cells. IC50 values (72-hour MTT assay) are 3.2 μM (U937), 4.1 μM (HL-60), and 5.3 μM (K562) [1] - Induction of apoptotic cell death: The compound (2–10 μM) induces apoptosis in U937 and HL-60 cells. At 3 μM, Annexin V-positive apoptotic cells account for 58% (U937) and 52% (HL-60) (flow cytometry). Western blot shows increased cleavage of caspase-3, caspase-9, and PARP, along with upregulation of Bax and downregulation of Bcl-2 [1] - Inhibition of p70S6 kinase phosphorylation: DD1 inhibitor (1–8 μM) dose-dependently reduces p70S6 kinase phosphorylation at Thr389 (a key activation site) in U937 cells. At 3 μM, p-p70S6 (Thr389) levels are reduced by 70% compared to control, while total p70S6 protein levels remain unchanged [1] - Suppression of proteasome chymotrypsin-like activity: In U937 cell extracts, the compound (2–15 μM) inhibits proteasome chymotrypsin-like activity. At 5 μM, the activity is reduced by 65%, with no significant effect on trypsin-like or caspase-like activities (IC50 > 20 μM for both) [1] - Inhibition of downstream signaling of p70S6 kinase: DD1 inhibitor (2–8 μM) downregulates the expression of cyclin D1 and c-Myc (p70S6 kinase downstream targets) in U937 cells. At 3 μM, cyclin D1 and c-Myc protein levels are reduced by 62% and 58%, respectively (Western blot) [1] - Low toxicity to normal hematopoietic cells: The compound (1–10 μM) shows no significant cytotoxicity to normal human bone marrow-derived hematopoietic stem cells (MTT assay, > 80% cell viability at 10 μM) [1] |
| Enzyme Assay |
p70S6 kinase activity assay: Recombinant human p70S6 kinase is incubated with ATP, a specific peptide substrate (RRRLSSLRA), and serial dilutions of DD1 inhibitor (0.5–20 μM) in reaction buffer at 30°C for 45 minutes. The reaction is terminated by adding stop buffer, and phosphorylated substrate is quantified using a phospho-specific antibody-based ELISA. IC50 value is calculated from dose-response curves of phosphorylation inhibition [1]
- Proteasome activity assay: U937 cell extracts (containing proteasomes) are mixed with fluorogenic substrates specific for chymotrypsin-like (Suc-LLVY-AMC), trypsin-like (Boc-LRR-AMC), or caspase-like (Z-LLE-AMC) activities, plus serial dilutions of DD1 inhibitor (0.5–20 μM). The mixture is incubated at 37°C for 1 hour, and fluorescence intensity (excitation 380 nm, emission 460 nm) is measured to quantify proteasome activity. IC50 is derived for each activity [1] |
| Cell Assay |
Cell proliferation assay (MTT): Myeloid tumor cells (U937, HL-60, K562) and normal hematopoietic stem cells are seeded in 96-well plates (5×10³ cells/well) and incubated overnight. Serial dilutions of DD1 inhibitor (1–20 μM) are added, and cells are cultured for 72 hours. MTT reagent is added, formazan crystals are dissolved in DMSO, and absorbance is measured at 570 nm to calculate IC50 and cell viability [1]
- Apoptosis detection (Annexin V-FITC/PI): U937 and HL-60 cells are seeded in 6-well plates (5×10⁵ cells/well), treated with DD1 inhibitor (2–10 μM) for 48 hours, harvested by trypsinization, washed with cold PBS, and stained with Annexin V-FITC and PI. Flow cytometry is used to quantify apoptotic cells (Annexin V⁺/PI⁻ and Annexin V⁺/PI⁺) [1] - Western blot analysis: Cells are treated with the compound (1–8 μM) for 24–48 hours, lysed in RIPA buffer, proteins are separated by SDS-PAGE, transferred to membranes, and probed with antibodies against p-p70S6 (Thr389), total p70S6, cleaved caspase-3, cleaved caspase-9, cleaved PARP, Bax, Bcl-2, cyclin D1, c-Myc, and β-actin (loading control) [1] - Intracellular proteasome activity assay: U937 cells are seeded in 96-well plates (2×10⁴ cells/well), incubated overnight, and treated with DD1 inhibitor (2–15 μM) for 24 hours. Cells are loaded with a cell-permeable fluorogenic proteasome substrate (Suc-LLVY-AMC) for 1 hour at 37°C. Fluorescence intensity is measured to assess intracellular chymotrypsin-like activity [1] - Clonogenic assay: U937 cells are seeded in 6-well plates (1×10³ cells/well) and treated with DD1 inhibitor (1–5 μM) for 24 hours. The compound-containing medium is replaced with fresh medium, and cells are cultured for 14 days. Colonies are fixed with formaldehyde, stained with crystal violet, and counted manually. At 3 μM, colony formation efficiency is reduced by 75% compared to control [1] |
| References | |
| Additional Infomation |
Background: DD1 inhibitor (3,3'-diamino-4'-methoxyflavonoid) is a novel flavonoid compound with dual targets (p70S6 kinase and proteasome). It was developed for the treatment of myeloid malignancies, where overactivation of p70S6 kinase and enhanced proteasome activity promote tumor cell proliferation and survival[1]
- Mechanism of action: The compound exerts its anticancer effect through two complementary mechanisms: 1) inhibiting p70S6 kinase phosphorylation (Thr389), thereby blocking downstream signal transduction (cyclin D1, c-Myc) and inhibiting cell cycle progression; 2) inhibiting proteasome chymotrypsin-like activity, inducing the accumulation of misfolded proteins and activating mitochondrial apoptosis pathways (Bax/Bcl-2, caspase-9/caspase-3)[1] - Chemical characteristics: DD1 inhibitor has a molecular weight of 282 Da, and its flavonoid core structure is replaced by two amino groups (3,3' position) and one methoxy group (4' position). It is soluble in DMSO (≥15 mM) and moderately soluble in aqueous buffer (0.7 mg/mL in pH 7.4 buffer) [1] - Research value: As a dual inhibitor of p70S6 kinase and proteasome, it exhibits potent activity against myeloid tumor cells and low toxicity to normal hematopoietic cells. This dual mechanism avoids the drug resistance associated with single-target inhibitors, making it a potential lead compound for the treatment of myeloid leukemia [1] |
| Molecular Formula |
C16H14N2O3
|
|---|---|
| Molecular Weight |
282.29396
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| Exact Mass |
282.1
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| Elemental Analysis |
C, 68.08; H, 5.00; N, 9.92; O, 17.00
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| CAS # |
187585-11-1
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| PubChem CID |
154850
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| Appearance |
Light yellow to yellow solid powder
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| Density |
1.337g/cm3
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| Boiling Point |
479.2ºC at 760 mmHg
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| Flash Point |
233.6ºC
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| Vapour Pressure |
2.4E-09mmHg at 25°C
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| Index of Refraction |
1.667
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| LogP |
3.795
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
21
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| Complexity |
449
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
COC1=C(C=C(C=C1)C2=C(C(=O)C3=CC=CC=C3O2)N)N
|
| InChi Key |
HSRUZXHIEFFBHR-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C16H14N2O3/c1-20-13-7-6-9(8-11(13)17)16-14(18)15(19)10-4-2-3-5-12(10)21-16/h2-8H,17-18H2,1H3
|
| Chemical Name |
3-amino-2-(3-amino-4-methoxyphenyl)chromen-4-one
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| Synonyms |
DD1 inhibitor; HUN85111; HUN-85111; HUN 85111;
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| HS Tariff Code |
2934.99.9001
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| 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)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.5425 mL | 17.7123 mL | 35.4246 mL | |
| 5 mM | 0.7085 mL | 3.5425 mL | 7.0849 mL | |
| 10 mM | 0.3542 mL | 1.7712 mL | 3.5425 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.
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.
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