Antitumor agent-101 is a selective covalent inhibitor of lysine methyltransferase G9a/GLP, with IC50s of 8.5 nM and 5.5 nM for G9a and GLP, respectively. Antitumor agent-101 displays antitumor effects in a PANC-1 xenograft model.

Physicochemical Properties

Molecular Formula	C26H38N6O3
Molecular Weight	482.62
CAS #	2848632-52-8
Appearance	Typically exists as solid at room temperature
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)

Biological Activity

Targets	G9a 8.5 nM (IC50) GLP 5.5 nM (IC50)
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.
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].
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.
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]
References	[1]. Feng Z, et.al. Structure-Based Design and Characterization of the Highly Potent and Selective Covalent Inhibitors Targeting the Lysine Methyltransferases G9a/GLP. J Med Chem. 2023 Jun 22;66(12):8086-8102.

Solubility Data

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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (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.