AFM-30a HCl is a potent protein arginine deiminase 2 (PAD2) inhibitor (antagonist) with excellent PAD2 selectivity. AFM-30a HCl binds to PAD2 with EC50 of 9.5 μM. AFM-30a HCl also inhibits H3 citrullination with EC50 of 0.4 μM. AFM-30a HCl may be used in research into certain cancers and a variety of autoimmune diseases such as rheumatoid arthritis (RA), multiple sclerosis, lupus and ulcerative colitis.

Physicochemical Properties

Molecular Formula	C24H28CLFN6O3
Molecular Weight	502.97
Related CAS #	AFM-30a;2095107-57-4
Appearance	White to off-white solid powder
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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.
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	EC50: 9.5 μM (PAD2); 0.4 μM (H3 citrullination)[1]
ln Vitro	Compound 30a, or AFM-30a hydrochloride, exhibits an EC50 of 9.5 μM and high efficacy to penetrate HEK293T/PAD2 cells and covalently alter PAD2[1]. With an EC50 of 0.4 μM, AFM-30a hydrochloride exhibits good capacity to block H3 citrullination[1].
ln Vivo	In PAD2-/-transgenic mice, AFM-30a hydrochloride reduces airway remodeling and inhibits NLRP3 signaling[2].
Cell Assay	Cell Viability Assay[1] Cell Types: HEK293T/PAD2 cells Tested Concentrations: Various concentrations Incubation Duration: 24 h Experimental Results: demonstrated low cytotoxicity for cells.
References	[1]. Development of a Selective Inhibitor of Protein Arginine Deiminase 2. J Med Chem. 2017 Apr 13;60(7):3198-3211. [2]. Role of PAD2 Regulated Inflammasome Signaling in Arsenic Induced Airway Inflammation and Remodeling. American Journal of Respiratory and Critical Care Medicine 2020;201:A2976.

Solubility Data

Solubility (In Vitro)

H2O :~50 mg/mL (~99.41 mM) DMSO :~50 mg/mL (~99.41 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 5 mg/mL (9.94 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 50.0 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: ≥ 5 mg/mL (9.94 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 50.0 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.9882 mL	9.9410 mL	19.8819 mL
	5 mM	0.3976 mL	1.9882 mL	3.9764 mL
	10 mM	0.1988 mL	0.9941 mL	1.9882 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.