HPK1-IN-3 is a potent and specific ATP-competitive hematopoietic progenitor kinase 1 (HPK1; MAP4K1) inhibitor (antagonist) with IC50 of 0.25 nM. HPK1-IN-3 has IL-2 cell potency with EC50 of 108 nM in human peripheral blood mononuclear cells (PBMC).

Physicochemical Properties

Molecular Formula	C23H22F4N6O2
Molecular Weight	490.453398227692
Exact Mass	490.174
CAS #	2739844-34-7
PubChem CID	155883041
Appearance	White to yellow solid powder
LogP	4.2
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	11
Rotatable Bond Count	6
Heavy Atom Count	35
Complexity	731
Defined Atom Stereocenter Count	0
InChi Key	RZLXOEIALAVGKH-UHFFFAOYSA-N
InChi Code	InChI=1S/C23H22F4N6O2/c1-33-7-6-12-9-18(35-2)17(8-13(12)11-33)30-22-29-10-14(20(28)34)21(32-22)31-19-15(23(25,26)27)4-3-5-16(19)24/h3-5,8-10H,6-7,11H2,1-2H3,(H2,28,34)(H2,29,30,31,32)
Chemical Name	4-[2-fluoro-6-(trifluoromethyl)anilino]-2-[(6-methoxy-2-methyl-3,4-dihydro-1H-isoquinolin-7-yl)amino]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

Targets	HPK1 0.25 nM (IC50)
ln Vitro	In human monocyte-derived dendritic cells, treatment with HPK1-IN-3 (Compound 27; 0.25–4 μM; 24 hours) significantly increased the concentration-dependent levels of pro-inflammatory cytokines TNF-α and IL-6 [1].
References	[1]. Discovery of Diaminopyrimidine Carboxamide HPK1 Inhibitors as Preclinical Immunotherapy Tool Compounds. ACS Med Chem Lett. 2021 Mar 19;12(4):653-661.

Solubility Data

Solubility (In Vitro)

DMSO: 83.33 mg/mL (169.91 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.24 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 20.8 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: ≥ 2.08 mg/mL (4.24 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 20.8 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	2.0389 mL	10.1947 mL	20.3894 mL
	5 mM	0.4078 mL	2.0389 mL	4.0779 mL
	10 mM	0.2039 mL	1.0195 mL	2.0389 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.