| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| 500mg | |||
| Other Sizes |
Purity: ≥98%
UNC0224 (UNC-0224) is a novel, potent and selective G9a histone methyltransferase inhibitor with anticancer activity. It inhibits G9a with IC50 of 15 nM in in the G9a Thioglo assay. In an ITC experiment to measure the binding affnityaffinity, UNC0224 showed 5-fold higher binding affinity than BIX01294. Through the first X-ray crystal structure of the G9a-UNC0224 complex, it was found that the 7-dimethy -laminopropoxy side chain of UNC0224 indeed occupied the lysine binding channel of G9a nicely, thus validating previous SAR experimental conclusions. In summary, UNC0224 is a significantly more potent G9a inhibitor than BIX-01294. The discovery of UNC0224 and the emergence of the first X-ray crystal structure of G9a with UNC0224 paved the way for studying the mechanism of G9a protein in vivo, which will enable structure-based design of novel inhibitors.
| Targets |
UNC0224 targets G9a histone lysine methyltransferase (EHMT2) and G9a-like protein (GLP/EHMT1) (G9a: IC50 = 15 nM for methyltransferase activity [1]
; GLP: IC50 = 190 nM for methyltransferase activity [1] ; no significant inhibition of other histone methyltransferases (EZH2, SUV39H1, SETDB1) with IC50 > 10 μM [1][2] |
|---|---|
| ln Vitro |
In the G9a ECSD and CLOT tests, UNC0224 (compound 10) had IC50 values of 43 nM and 57 nM, respectively. In the GLP ECSD and CLOT tests, UNC0224's IC50 values were 50 nM and 58 nM, respectively. The Kd value of UNC0224 for the G9a protein in the ITC experiment was 23 nM[2].
1. UNC0224 potently inhibited recombinant G9a methyltransferase activity with an IC50 of 15 nM, and showed 12.7-fold selectivity for G9a over GLP (IC50 = 190 nM) in peptide-based methyltransferase assays [1] 2. In HeLa cells, UNC0224 (0.1-5 μM) dose-dependently reduced global H3K9 dimethylation (H3K9me2) levels by up to 80% at 1 μM, as measured by western blot; no significant changes in H3K4me2, H3K9me3, or H3K27me3 were observed, confirming epigenetic selectivity [1] 3. Structure-activity relationship (SAR) studies in [2] demonstrated that the 7-aminoalkoxy side chain of UNC0224 is critical for G9a binding; modification of the quinazoline core or substitution of the diamino group reduced potency (IC50 > 100 nM for G9a) [2] 4. UNC0224 showed no cytotoxicity in HeLa and NIH/3T3 cells at concentrations up to 10 μM, with cell viability >95% after 72-hour treatment [2] |
| Enzyme Assay |
1. G9a/GLP methyltransferase activity assay: Recombinant G9a or GLP catalytic domain protein was incubated with a biotinylated histone H3 (1-21) peptide substrate, S-adenosylmethionine (SAM), and serial dilutions of UNC0224 (0.001-10 μM) in reaction buffer at 30°C for 60 minutes; methylated peptide product was detected using a europium-labeled anti-methyl-H3K9 antibody and time-resolved fluorescence resonance energy transfer (TR-FRET); dose-response curves were generated to calculate IC50 values for enzyme inhibition [1]
2. Selectivity assay for other methyltransferases: Recombinant EZH2, SUV39H1, and SETDB1 proteins were incubated with their respective peptide substrates, SAM, and UNC0224 (0.01-10 μM) under identical conditions to the G9a assay; methyltransferase activity was quantified by TR-FRET, and percentage inhibition was calculated to assess off-target effects [2] |
| Cell Assay |
1. H3K9me2 epigenetic modification assay: HeLa cells were seeded in 6-well plates at a density of 5×10⁵ cells/well and treated with UNC0224 (0.1, 0.5, 1, 5 μM) for 24 hours; histone extracts were prepared using acid extraction, and equal amounts of protein were separated by SDS-PAGE; western blot was performed with antibodies against H3K9me2, H3K4me2, H3K9me3, and total H3 (loading control); band intensities were quantified by densitometry to measure changes in methylation levels [1]
2. Cell viability assay: HeLa and NIH/3T3 cells were seeded in 96-well plates at 5×10³ cells/well and treated with UNC0224 (0.01-10 μM) for 72 hours at 37°C with 5% CO₂; a colorimetric cell viability reagent was added, and absorbance was measured at 490 nm; cell viability was calculated relative to vehicle-treated controls [2] |
| Toxicity/Toxicokinetics |
1. At concentrations up to 10 μM, UNC0224 did not exhibit significant cytotoxicity to mammalian cell lines (HeLa, NIH/3T3), and cell viability was >95% after 72 hours of exposure [2].
|
| References |
|
| Additional Infomation |
1. UNC0224 is a first-generation 2,4-diamino-7-aminoalkoxyquinazoline derivative and one of the most effective and selective G9a inhibitors reported at the time.[1] 2. The mechanism of action of UNC0224 is to bind to the SAM binding pocket of the G9a catalytic domain and compete with SAM for binding sites, thereby blocking the transfer of methyl groups to histone H3.[1] 3. The structure-activity relationship study in [2] identified the key structural features of UNC0224 that enable it to exert G9a inhibitory activity: a 7-(3-dimethylaminopropoxy) side chain, an unsubstituted 2,4-diamino group on the quinazoline ring, and a planar aromatic core.[2] 4. UNC0224 is a valuable tool compound for in vitro studies of G9a-mediated epigenetic regulation, but its development still needs further research. Limited by poor water solubility and lack of in vivo characterization [1][2]
|
| Molecular Formula |
C26H43N7O2
|
|---|---|
| Molecular Weight |
485.66532
|
| Exact Mass |
485.347
|
| CAS # |
1197196-48-7
|
| PubChem CID |
44251522
|
| Appearance |
White to yellow solid powder
|
| Density |
1.1±0.1 g/cm3
|
| Boiling Point |
641.0±65.0 °C at 760 mmHg
|
| Flash Point |
341.5±34.3 °C
|
| Vapour Pressure |
0.0±1.9 mmHg at 25°C
|
| Index of Refraction |
1.589
|
| LogP |
1.3
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
9
|
| Rotatable Bond Count |
9
|
| Heavy Atom Count |
35
|
| Complexity |
619
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
XIVUGRBSBIXXJE-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C26H43N7O2/c1-30(2)10-7-17-35-24-19-22-21(18-23(24)34-5)25(27-20-8-13-32(4)14-9-20)29-26(28-22)33-12-6-11-31(3)15-16-33/h18-20H,6-17H2,1-5H3,(H,27,28,29)
|
| Chemical Name |
7-[3-(dimethylamino)propoxy]-2-(hexahydro-4-methyl-1H-1,4-diazepin-1-yl)-6-methoxy-N-(1-methyl-4-piperidinyl)-4-quinazolinamine
|
| Synonyms |
UNC-0224; UNC0224; UNC 0224
|
| 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 (In Vitro) |
DMSO : ~16.67 mg/mL (~34.32 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.67 mg/mL (3.44 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 16.7 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: ≥ 1.67 mg/mL (3.44 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 16.7 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. View More
Solubility in Formulation 3: ≥ 1.67 mg/mL (3.44 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0590 mL | 10.2951 mL | 20.5901 mL | |
| 5 mM | 0.4118 mL | 2.0590 mL | 4.1180 mL | |
| 10 mM | 0.2059 mL | 1.0295 mL | 2.0590 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA