AFM32a (PAD2-IN-1) HCl, a benzimidazole-based analogue, is a potent and specific inhibitor of protein arginine deiminase 2 (PAD2). AFM32a HCl is more selective for PAD2 than PAD4 (95-fold) and PAD3 (79-fold).

Physicochemical Properties

Molecular Formula	C25H30CLFN6O3
Molecular Weight	517
CAS #	2988594-85-8
Related CAS #	AFM32a;2095109-82-1
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	Protein Arginine Deiminase 2 (PAD2)[1]
ln Vitro	Compound 32a, or AFM32a hydrochloride, has an EC 50 of 8.3 μM in HEK293T/PAD2 cells in the target engagement experiment. This indicates that AFM32a's improved potency outweighs its relatively weak cell entry ability[1]. With an EC 50 of 2.7 μM, AFM32a (compound 32a; 1-25 μM) hydrochloride treatment substantially suppresses histone H3 citrullination in HEK293T/PAD2 cells[1].
References	[1]. Development of a Selective Inhibitor of Protein Arginine Deiminase 2. J Med Chem. 2017 Apr 13;60(7):3198-3211.

Solubility Data

Solubility (In Vitro)

DMSO :~100 mg/mL (~193.42 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.84 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 25.0 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: ≥ 2.5 mg/mL (4.84 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 25.0 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (4.84 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.9342 mL	9.6712 mL	19.3424 mL
	5 mM	0.3868 mL	1.9342 mL	3.8685 mL
	10 mM	0.1934 mL	0.9671 mL	1.9342 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.