DB818 dihydrochloride
Based on 1 Customer Validation
DB818 dihydrochloride is a HOXA9 transcription factor inhibitor with an IC50 of 0.2-0.3 µM. DB818 dihydrochloride inhibits HOXA9 DNA binding and transcription, binding to AT-rich minor grooves with a Kd of 4.6 nM. DB818 dihydrochloride enters the nucleus and induces cancer cell apoptosis, cell cycle arrest, and granulocytic and monocytic differentiation, while inhibiting proliferation, colony formation, and leukemic burden. DB818 dihydrochloride downregulates MYB and BCL2, upregulates FOS, and weakly downregulates HOXA9 protein. DB818 dihydrochloride has anti-Babesia activity. DB818 dihydrochloride can be used in research related to acute myeloid leukemia, non-small cell lung cancer, and babesiosis.
For research use only. We do not sell to patients.
- Purity : 99.99%
- Formula: C19H18Cl2N6S
- Molecular Weight:433.36
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Storage:
-20°C, sealed storage, away from moisture
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
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Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Protocols
- Purity & Documentation
- References
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Biological Activity
Description
Cellular Effect
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Cell Line
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Type | Value | Description | References |
|---|---|---|---|---|
| MigA9 | IC50 |
9.97 μM
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Antiproliferative activity against murine MigA9 leukemia cells assessed as reduction in cell survival after 72 hrs by MTS assay.
Antiproliferative activity against murine MigA9 leukemia cells assessed as reduction in cell survival after 72 hrs by MTS assay.
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30645099 |
| L6 | IC50 |
10.6 μg/mL
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Cytotoxicity against rat L6 skeletal myoblasts by Alamar blue assay.
Cytotoxicity against rat L6 skeletal myoblasts by Alamar blue assay.
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21537025 |
In Vitro
DB818 (1-50 μM; 96 h) dihydrochloride sensitivity in primary AML blasts positively correlates with HOXA9 expression, and the NPM1 and MLL mutant subtypes exhibit higher sensitivity[1].
DB818 (20 μM; 48 h) dihydrochloride selectively inhibits HOXA9-dependent transcription in K562 cells without affecting K562 survival[2].
DB818 (2.5 μM; 10 days) dihydrochloride reduces the clonogenic capacity of MigA9 cells and shifts colony subtypes toward granulocyte/monocyte differentiation, but has no such effect in HOXA9-negative K562 cells[2].
DB818 (10-20 μM; 24-48 h) dihydrochloride inhibits the binding of HOXA9 to its consensus DNA oligonucleotide in OCI/AML3, MV4-11, and THP-1 cells[4].
DB818 (10-20 μM; 96 h) dihydrochloride shows stronger inhibition of the growth of OCI/AML3, MV4-11, and THP-1 AML cell lines than of other leukemia cell lines, and has a slight inhibitory effect on normal lymphocytes[4].
DB818 (2.5-5 μM; 6-24 h) dihydrochloride reverses the HOXA9-related transcriptional program in MigA9 cells and induces changes in the expression of differentiation/apoptosis-related genes[2].
DB818 dihydrochloride increases the annexin V-positive rate in MV4-11, OCI/AML3, and THP-1 cells and upregulates CD11b in THP-1 cells[4].
DB818 (10-20 μM; 24-48 h) dihydrochloride downregulates MYB and MYC in OCI/AML3, MV4-11, and THP-1 cells, and slightly reduces BCL2 and HOXA9 in OCI/AML3 cells[4].
DB818 (20 μM; 24 h) dihydrochloride alters the expression of HOXA9 target genes in OCI/AML3, MV4-11, and THP-1 cells, downregulating BCL2 and MYB and upregulating FOS[4].
DB818 (5-15 μM, 3-7 days) dihydrochloride induces features of monocyte-to-macrophage differentiation in THP-1 cells[1].
DB818 (5-10 μM; 48 h) dihydrochloride induces the expression of granulocyte/monocyte differentiation-related genes, including CD11b and CD14, in MigA9 cells[2].
DB818 dihydrochloride does not significantly promote differentiation in normal human CD34+ lineage differentiation assays, and its toxicity is lower than that of AraC[1].
DB818 dihydrochloride inhibits the binding of HOXA9 to its cognate HBS DNA in a cell-free ELISA-derived assay[2].
DB818 dihydrochloride is a potent inhibitor of HOXA9 binding to HBS and Akap13-HBS DNA, as confirmed by EMSA[2].
DB818 dihydrochloride selectively recognizes the 5′-ATTTA sequence within the HOXA9 homeodomain binding site[2].
DB818 (1 nM-1 μM; 3 min) dihydrochloride binds the HOXA9 homologous HBS sequence with a KD of 4.6 nM, showing approximately 75-fold selectivity over the mutant HBS[2].
DB818 (20 μM; 2 h) dihydrochloride localizes to the nucleus of MigA9 leukemia cells[2].
DB818 dihydrochloride binds the AATT minor groove of d (CGCGAATTCGCG)2 and forms direct and water-mediated hydrogen-bond contacts in the PDB 1VZK complex[3].
DB818 dihydrochloride exhibits cytotoxicity against L6 rat skeletal muscle myoblasts, with an IC50 of 10.6 μg/mL[6].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Cell Line:THP-1 cells; EOL-1 and U937 cells
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Concentration:5, 10, 15 μM
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Incubation Time:3 or 6 days (5 μM); 72 h (5 μM); 7 days (5, 10, 15 μM)
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Result:Induced morphological changes including reduced nucleus-to-cytoplasm ratio, basophilic cells, phagocytic vacuoles, and pseudopods.
CD11b surface expression increased at 7 days, starting on day 2; CD14 increased later at day 4; CD11b/CD14 double-positive cells increased dose-dependently at 5, 10, and 15 μM.
Differentiation was also induced in EOL-1 and U937 cells.
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Cell Line:THP-1 AML cell line; U937, EOL-1, MV4-11, SHI-1, and HOXA9-independent leukemia cells
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Concentration:10 μM (proliferation); 1.5, 2.5 and 5 μM (clonogenicity); 5, 10 and 15 μM (cell death/apoptosis)
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Incubation Time:days indicated (proliferation); 14 days (clonogenicity); 7 days (cell death/apoptosis)
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Result:Decreased proliferation, reduced colony formation, increased PI-positive dead cells, and increased AnnexinV/PI double-positive apoptotic cells.
Significantly decreased colony number and size in HOXA9-positive U937, EOL-1, MV4-11, and SHI-1 cell lines, while HOXA9-independent leukemia cells remained unaffected.
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Cell Line:Hoxa9-expressing murine MigA9 leukemia cells
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Concentration:10, 15, and 20 μM
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Incubation Time:24 and 48 h
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Result:PI-positive cells at 24 and 48 h: 10 μM = 6.95% and 29.40%; 15 μM = 13.50% and 64.03%; 20 μM = 21.07% and 75.53%.
Sub-G1 at 48 h: 10 μM = 9.46%; 15 μM = 14.58%; 20 μM = 12.00%.
Cell cycle at 48 h: 10 μM = G0/G1 50.38%, S 35.42%, G2/M 14.19%; 15 μM = G0/G1 58.30%, S 14.80%, G2/M 26.90%; 20 μM = G0/G1 56.02%, S 9.25%, G2/M 34.73%.
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Cell Line:Murine MigA9 leukemia cells
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Concentration:5 and 10 μM
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Incubation Time:48 h
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Result:Increased expression of Itgam (CD11b), Oasl2, Akap13, Arfgef1, and CD14.
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Cell Line:human AML cell lines OCI/AML3, MV4-11, THP-1, other AML/ALL cell lines, and normal lymphocytes
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Concentration:10, 20 μM
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Incubation Time:96 h
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Result:Suppressed the growth of five AML and two ALL cell lines in a dose-dependent manner.
MV4-11, THP-1, and OCI/AML3 cells were more susceptible than the other cell lines.
Slightly inhibited the growth of normal lymphocytes from two out of three volunteers.
Time-course curves confirmed growth suppression of OCI/AML3, MV4-11, and THP-1 cells.
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Cell Line:human AML cell lines OCI/AML3, MV4-11, and THP-1
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Concentration:10 and 20 μM
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Incubation Time:24 and 48 h
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Result:Down-regulated MYB and MYC protein expression in OCI/AML3, MV4-11, and THP-1 cells and slightly reduced BCL2 in OCI/AML3 cells.
Also weakly down-regulated HOXA9 expression.
In Vivo
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:NSG (NOD.Cg-Prkdcscid-Il2rgtm1Wjl/SzJ; 6-8 weeks old)[1]
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Dosage:5 mg/kg
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Administration:i.p.
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Result:Bioluminescent THP-1-Luc signal decreased over time relative to vehicle.
Spleen weight was decreased.
The number of THP-1 (hCD45-positive) cells per spleen was decreased.
The percentage and/or median expression of hCD11b-positive THP-1 cells was increased in bone marrow, liver, blood, and ascites.
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Animal Model:NSG (NOD.Cg-Prkdcscid-Il2rgtm1Wjl/SzJ)[1]
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Dosage:5 mg/kg
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Administration:i.p.
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Result:Reduced splenomegaly associated with U937 xenograft.
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Animal Model:Swiss NMRI (female, 18-20 g)[6]
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Dosage:25 mg/kg; 12.5 mg/kg
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Administration:s.c.; at 4, 24, 48, and 72 h postinfection; 4 days
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Result:At 25 mg/kg s.c. for 4 days, cured 3/3 mice.
At 12.5 mg/kg s.c. for 4 days, all mice showed recrudescence around day 20 postinfection.
- Nuclear Protein Extraction (High-Salt/Hypotonic Fractionation)
- Protocol for Electrophoretic Mobility Shift Assay (EMSA)
- Detection of Apoptosis
- TUNEL staining for apoptotic DNA fragmentation
- Flow cytometric DNA-content cell-cycle staining
- Annexin V plus membrane-impermeant dye apoptosis staining
- BrdU Incorporation Assay
- Apoptosis Solutions
- Protocol for Cell Cycle
- Soft Agar Colony Formation Assay
- Colony Formation (Clonogenic) Assay
- Cell Viability Determination by MTT Colorimetric Assay
Chemical Information
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Appearance Solid
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Molecular Weight 433.36
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Formula C19H18Cl2N6S
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Color Off-white to yellow
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SMILES
N=C(C1=CC=C(C2=CC=C(C3=NC4=CC=C(C(N)=N)C=C4N3)S2)C=C1)N.Cl.Cl
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
-20°C, sealed storage, away from moisture
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
Solvent & Solubility
In Vitro:
DMSO : 15 mg/mL (34.61 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
H2O : 5 mg/mL (11.54 mM; Need ultrasonic and warming)
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 (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.
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 (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Working solution concentration: 0.22 mg/mL
Purity & Documentation
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Data Sheet (300 KB)
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SDS (251 KB)
- English - EN (251 KB)
- Français - FR (251 KB)
- Deutsch - DE (251 KB)
- Norwegian - NO (251 KB)
- Español - ES (251 KB)
- Swedish - SV (251 KB)
- Italian - IT (251 KB)
- Korean - KR (251 KB)
- Portuguese - PT (251 KB)
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Handling Instructions (2659 KB)
References
[1]. Lambert M, et al. Induction of AML cell differentiation using HOXA9/DNA binding inhibitors as a potential therapeutic option for HOXA9-dependent AML. HemaSphere. 2024 May;8(5):e77. [Content Brief]
[2]. Depauw S, et al. Heterocyclic Diamidine DNA Ligands as HOXA9 Transcription Factor Inhibitors: Design, Molecular Evaluation, and Cellular Consequences in a HOXA9-Dependant Leukemia Cell Model. Journal of medicinal chemistry. 2019 Feb 14;62(3):1306-1329. [Content Brief]
[4]. Sonoda Y, et al. Effects of HOXA9 Inhibitor DB818 on the Growth of Acute Myeloid Leukaemia Cells. Anticancer research. 2021 Apr;41(4):1841-1847. [Content Brief]
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 (sealed storage, away from moisture). 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 |
|---|---|---|---|---|---|
| H2O / DMSO | 1 mM | 2.3076 mL | 11.5378 mL | 23.0755 mL | 57.6888 mL |
| 5 mM | 0.4615 mL | 2.3076 mL | 4.6151 mL | 11.5378 mL | |
| 10 mM | 0.2308 mL | 1.1538 mL | 2.3076 mL | 5.7689 mL | |
| DMSO | 15 mM | 0.1538 mL | 0.7692 mL | 1.5384 mL | 3.8459 mL |
| 20 mM | 0.1154 mL | 0.5769 mL | 1.1538 mL | 2.8844 mL | |
| 25 mM | 0.0923 mL | 0.4615 mL | 0.9230 mL | 2.3076 mL | |
| 30 mM | 0.0769 mL | 0.3846 mL | 0.7692 mL | 1.9230 mL |
* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.
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