| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
G9a 8.5 nM (IC50) GLP 5.5 nM (IC50)
G9a (lysine methyltransferase); GLP (lysine methyltransferase); H3K9me2 (histone H3 lysine 9 dimethylation); PANC-1 pancreatic cancer cells; MDA-MB-231 breast cancer cells; Cell proliferation; Colony formation |
|---|---|
| ln Vitro |
Compound 27 (Antitumor Agent-101; 0-5 μM; 48 hours) suppresses PANC-1 and MDA-MB-231 cell proliferation and colony formation [1]. H3K9me2 levels in PANC-1 and MDA-MB-231 cells are efficiently reduced by antitumor agent-101 (0-10 μM; 0-96 hours) in a concentration- and time-dependent manner.
Antitumor agent-101 (Compound 27) (0-5 μM; 48 hours) significantly inhibits proliferation and colony formation of PANC-1 and MDA-MB-231 cells with IC50 values of 2.68 and 2.88 μM, respectively. It effectively reduces H3K9me2 in PANC-1 and MDA-MB-231 cells in a concentration- and time-dependent manner (0-10 μM; 0-96 hours). The compound significantly suppresses colony formation in MDA-MB-231 and PANC-1 cell lines at 2.5 μM. |
| ln Vivo |
Compound 27 (Antitumor Agent-101), administered intraperitoneally five days a week at a dose of 2 mg/kg, stops the formation of PANC-1 xenograft tumors by blocking G9a/GLP methyltransferase activity [1]. The mean residence time (MRT) of antitumor agent-101 (2 mg/kg, i.p.) was 0.61 hours, and its Cmax was 316 ng/mL [1].
In vivo, Antitumor agent-101 shows antitumor efficacy in the PANC-1 xenograft model. The compound is a selective covalent inhibitor of G9a/GLP and demonstrates target engagement in vivo as evidenced by reduced H3K9me2 levels in tumor tissues. Further pharmacokinetic characterization is needed to establish its absorption, distribution, metabolism, and excretion profile. |
| Enzyme Assay |
For G9a/GLP inhibition assays: incubate recombinant G9a or GLP with substrate peptide (e.g., H3 peptide) and S-adenosylmethionine (SAM) in assay buffer. Add Antitumor agent-101 at varying concentrations (0.1 nM to 10 μM) and measure methyltransferase activity by scintillation proximity assay or fluorescence-based detection. Calculate IC50 by nonlinear regression.
|
| Cell Assay |
Cell Proliferation Assay[1]
Cell Types: PANC-1 and MDA-MB-231 cells Tested Concentrations: 0, 1.25, 2.5, 5 μM Incubation Duration: 48 hrs (hours) Experimental Results: Ihibited proliferation of PANC-1 and MDA-MB-231 cells with IC50s of 2.68 and 2.88 μM, respectively. Dramatically suppressed the colony formation in MDA-MB-231 and PANC-1 cell lines at 2.5 μM. Western Blot Analysis[1] Cell Types: PANC-1 and MDA-MB-231 cells Tested Concentrations: 2.5, 5, 10 μM Incubation Duration: 4, 48, 72, or 96 hrs (hours) Experimental Results: Effectively decreased H3K9me2 in PANC -1 and MDA-MB-231 cells in a concentration- and time-dependent manner. Still Dramatically inhibited the levels of H3K9me2 in the cells treated with compound 27 were still Dramatically inhibited within 24h after Antitumor agent-101 was washed out, and the levels of H3K9me2 were recovered after 48h. For antiproliferative assays: seed PANC-1 and MDA-MB-231 cells in 96-well plates and treat with Antitumor agent-101 at concentrations of 0, 1.25, 2.5, and 5 μM for 48 hours. Assess cell viability by CCK-8 assay. For colony formation: seed cells at low density and treat with compound (2.5 μM) for 10-14 days, then stain with crystal violet and count colonies. For H3K9me2 detection: use Western blot with specific antibody. |
| Animal Protocol |
Animal/Disease Models: PANC-1 xenograft tumor models in male Balb/c nu/nu (nude) mice[1]
Doses: 2 mg/kg Route of Administration: intraperitoneal (ip)injection (ip ), 5 days a week (5 days on and 2 days off). Experimental Results: demonstrated potent antitumor activity with a tumor growth inhibition (TGI) rate of 52.2% with no obvious toxicity. demonstrated lower levels of H3K9me2 than the vehicle group. Animal/Disease Models: Male ICR Mice (pharmacokinetic/PK assay)[1] Doses: 2 mg/kg Route of Administration: intraperitoneal (ip)injection Experimental Results: pharmacokinetic/PK parameters for antitumor agent-101 (Compound 27) in rats [1] For in vivo studies: use a murine xenograft model with PANC-1 cells implanted subcutaneously into immunodeficient mice. When tumors reach 100-200 mm³, administer Antitumor agent-101 by intraperitoneal injection or oral gavage at doses ranging from 5-50 mg/kg daily for 2-3 weeks. Measure tumor volume twice weekly. At sacrifice, harvest tumors for H3K9me2 detection by Western blot and immunohistochemistry. For pharmacokinetic studies: collect plasma at various time points for LC-MS/MS analysis. |
| References |
| Molecular Formula |
C26H38N6O3
|
|---|---|
| Molecular Weight |
482.62
|
| Exact Mass |
482.301
|
| CAS # |
2848632-52-8
|
| PubChem CID |
168476152
|
| Appearance |
Typically exists as solid at room temperature
|
| tPSA |
83.1
|
| Hydrogen Bond Donor Count |
1
|
| Rotatable Bond Count |
10
|
| Heavy Atom Count |
35
|
| Complexity |
682
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CN1CCC(CC1)NC2=NC(=NC3=CC(=C(C=C32)OC)OCCCN4CCCC4)N(C)C(=O)C=C
|
| InChi Key |
GHCDVBSKNGUGCK-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C26H38N6O3/c1-5-24(33)31(3)26-28-21-18-23(35-16-8-13-32-11-6-7-12-32)22(34-4)17-20(21)25(29-26)27-19-9-14-30(2)15-10-19/h5,17-19H,1,6-16H2,2-4H3,(H,27,28,29)
|
| Chemical Name |
N-[6-methoxy-4-[(1-methylpiperidin-4-yl)amino]-7-(3-pyrrolidin-1-ylpropoxy)quinazolin-2-yl]-N-methylprop-2-enamide
|
| 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) |
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
|
|---|---|
| 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 | 2.0720 mL | 10.3601 mL | 20.7202 mL | |
| 5 mM | 0.4144 mL | 2.0720 mL | 4.1440 mL | |
| 10 mM | 0.2072 mL | 1.0360 mL | 2.0720 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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