PRMT5-IN-19 (Compound 41) is an orally bioactive, non-nucleoside PRMT5 selective inhibitor (antagonist) with IC50s of 23.9 nM (radioactive biochemical assay), 47 nM (AlphaLISA assay). PRMT5-IN-19 can occupy the SAM-binding pocket in PRMT5 and block the methyltransferase activity, with better selectivity than PRMTs and PKMTs. PRMT5-IN-19 inhibits cell growth/proliferation by inducing apoptosis and may be utilized in cancer-related research.

Physicochemical Properties

Molecular Formula	C25H24N4O
Molecular Weight	396.48
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

ln Vitro	Compound 41, PRMT5-IN-19, exhibits potent anti-proliferative properties against the A375 cell, as evidenced by its IC50 value of 1.36 μM[1]. Compared to other histone methyltransferases (PRMT1 and PRMT4) and PKMTS (EZH2, NSD2, MLL1, and MLL4), PRMT5-IN-19 has better selectivity for PRMT5 (IC50 value of 23.9 nM) [1]. The SAM-binding pocket in PRMT5 is where PRMT5-IN-19 binds[1]. Multiple cancer cell lines (A-375, CHL-1, SNU-423, SNU-449, MDA-MB-231, MDA-MA-453, MV-4-11, MOLM13) are inhibited in their growth by PRMT5-IN-19 (4-5 days), with an IC50 value ranging from 1.08 to 3.45 μM[1]. In A375 cells, PRMT5-IN-19 inhibits arginine symmetrical dimethylation [1]. Apoptosis is induced in a concentration-dependent manner by PRMT5-IN-19 (0–4 μM, 48 h), which reduces the growth of A375 cells[1].
ln Vivo	With favorable PK properties and considerable anticancer activity, PRMT5-IN-19 (Compound 41, A375 xenograft model, 75 mg/kg/d, po, 19 days) does not cause noticeable toxicity or apparent body weight loss[1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: A-375, CHL-1, SNU-423, SNU-449, MDA-MB-231, MDA-MA-453, MV-4- 11, MOLM13 Tested Concentrations: 0-10 μM Incubation Duration: 5 days Experimental Results: Inhibited proliferation of multiple cancer cell lines with IC50 value ranging from 1.08 to 3.45 μM. Western Blot Analysis[1] Cell Types: A-375 cells Tested Concentrations: 0.5, 1, 2, 4,8 μM Incubation Duration: 48 h. Experimental Results: Inhibited arginine symmetrical dimethylation in a dose-dependent manner.
Animal Protocol	Animal/Disease Models: A375 cell-derived nude mouse xenograft model[1]. Doses: 75 mg/kg/d Route of Administration: Po, 19 days Experimental Results: Had no effect on the body weight, displayed antitumor efficacy with a tumor growth inhibition (TGI) rate of 73% by inhibiting the methyltransferase activity of PRMT5. Animal/Disease Models: Rats and mice[1]. Doses: 10 mg/kg for po, 3 mg/kg for iv Route of Administration: Po, iv (pharmacokinetic/PK Analysis) Experimental Results: pharmacokinetic/PK parameters for PRMT5 -IN-19 in SD Rats and Micea,c [1].
References	[1]. Deqin Rong, et al. Structure-Aided Design, Synthesis, and Biological Evaluation of Potent and Selective Non-Nucleoside Inhibitors Targeting Protein Arginine Methyltransferase 5. J Med Chem. 2022 Jun 9;65(11):7854-7875.

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	2.5222 mL	12.6110 mL	25.2220 mL
	5 mM	0.5044 mL	2.5222 mL	5.0444 mL
	10 mM	0.2522 mL	1.2611 mL	2.5222 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.