DY-46-2 is a highly efficient and selective novel non-nucleoside DNA methyltransferase 3A (DNMT3A) inhibitor (antagonist) with IC50 of 1.3 μM.

Physicochemical Properties

Molecular Formula	C19H22N6O5S
Molecular Weight	446.480182170868
Exact Mass	446.137
CAS #	1105110-83-5
PubChem CID	71035751
Appearance	White to off-white solid powder
LogP	-0.1
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	6
Heavy Atom Count	31
Complexity	824
Defined Atom Stereocenter Count	0
SMILES	C1N(CCN2CCCC2)C(=O)C2C(C(NC3=CC=C(S(N)(=O)=O)N=C3)=O)=C(C)OC=2N=1
InChi Key	FAZNWIGAOYXZGW-UHFFFAOYSA-N
InChi Code	InChI=1S/C19H22N6O5S/c1-12-15(17(26)23-13-4-5-14(21-10-13)31(20,28)29)16-18(30-12)22-11-25(19(16)27)9-8-24-6-2-3-7-24/h4-5,10-11H,2-3,6-9H2,1H3,(H,23,26)(H2,20,28,29)
Chemical Name	6-methyl-4-oxo-3-(2-pyrrolidin-1-ylethyl)-N-(6-sulfamoylpyridin-3-yl)furo[2,3-d]pyrimidine-5-carboxamide
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

ln Vitro	DY-46-2 exhibits DNMT3A inhibitory activity with an IC50 value of 0.39 μM, increasing linearly with DNA concentration (IDT-01) [1]. It is available at concentrations ranging from 0.1 to 100 μM for 24, 48, and 72 hours. DY-46-2 (0.1-100 μM, 24, 48, and 72 h) exhibits inhibitory effect against G9a, DNMT1, and DNMT3B, with IC50 values of >500 μM, 105 μM, and 13.0 μM, respectively [1]. For HCT116, U937, K562, A549, DU145, and 72-hour exposure, DY-46-2 (0.1-100 μM, 24, 48, and 72 hours) had IC50 values of 0.7 μM, 0.3 μM, 0.7 μM, 0.5 μM, 2.1 μM, and 1.7 μM, and for THP 91 μM -1, respectively. Peripheral blood mononuclear cells (PBMCs) are not cytotoxic to DY-46-2 (0.1-100 μM, 24, 48, and 72 hours) and it effectively suppresses the proliferation of cancer cells [1]. In HCT116 cells, DY-46-2 (1 μM) dramatically lowers the reactive expression of silenced TSG (p53) and DNMT3A protein levels [1].
Cell Assay	Cell Viability Assay[1] Cell Types: THP-1, HCT116, U937, K562, A549, DU145 and PBMC Cell Tested Concentrations: 0.1-100 μM Incubation Duration: 24, 48 and 72 hrs (hours) Experimental Results: Significant across the dose range Inhibited potency and time-dependent manner and was not cytotoxic in non-tumor PBMC (IC50 >100 μM). Cell proliferation assay[1] Cell Types: THP-1, HCT116, U937, K562, A549, DU145 and PBMC Cell Tested Concentrations: 0.1-100 μM Incubation Duration: 24, 48 and 72 hrs (hours) Experimental Results: demonstrated high anti-proliferative activity against All cancer cells have cytotoxicity in the micromolar range. Western Blot Analysis [1] Cell Types: HCT116 Cell Tested Concentrations: 1 μM Incubation Duration: 72 hrs (hours) Experimental Results: DNMT3A and p53 protein levels were diminished in HCT116 cells, apparently sufficient to reactively express silenced TSG (p53).
References	[1]. Discovery of novel non-nucleoside inhibitors with high potency andselectivity for DNA methyltransferase 3A. European Journal of Medicinal Chemistry (2022).

Solubility Data

Solubility (In Vitro)

DMSO : ~50 mg/mL (~111.99 mM)

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	2.2397 mL	11.1987 mL	22.3974 mL
	5 mM	0.4479 mL	2.2397 mL	4.4795 mL
	10 mM	0.2240 mL	1.1199 mL	2.2397 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.