GNE-1858 is a potent ATP-competitive hematopoietic progenitor cell kinase-1 (HPK1) inhibitor, with IC50s of 1.9 nM, 1.9 nM, and 4.5 against wild-type and active mimic mutant strains HPK1-TSEE and HPK1-SA, respectively. nM.

Physicochemical Properties

Molecular Formula	C21H26F2N8
Molecular Weight	428.48
Exact Mass	428.224
CAS #	2680616-96-8
PubChem CID	146026186
Appearance	Light yellow to yellow solid powder
LogP	2.7
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	5
Heavy Atom Count	31
Complexity	622
Defined Atom Stereocenter Count	0
SMILES	C1=NC(NC2C=C(C3CCNC3)N=C(N3CCC(F)(F)C3)N=2)=CC2N(C(C)C)N=CC1=2
InChi Key	JKPAJCPZQIJPPS-UHFFFAOYSA-N
InChi Code	InChI=1S/C21H26F2N8/c1-13(2)31-17-8-18(25-10-15(17)11-26-31)28-19-7-16(14-3-5-24-9-14)27-20(29-19)30-6-4-21(22,23)12-30/h7-8,10-11,13-14,24H,3-6,9,12H2,1-2H3,(H,25,27,28,29)
Chemical Name	N-[2-(3,3-difluoropyrrolidin-1-yl)-6-pyrrolidin-3-ylpyrimidin-4-yl]-1-propan-2-ylpyrazolo[4,3-c]pyridin-6-amine
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	Hematopoietic progenitor kinase-1 (HPK1) is a type of protein kinase that is linked to Ste20 and is expressed only in the hematopoietic compartment, which includes T cells, B cells, and dendritic cells. It is a serine/threonine kinase. HPK1 inhibits T cell proliferation and effector function as a negative-feedback regulator of T cell receptor signaling[1].
References	[1]. Hematopoietic Progenitor Kinase-1 Structure in a Domain-Swapped Dimer. Structure. 2019 Jan 2;27(1):125-133.e4.

Solubility Data

Solubility (In Vitro)

DMSO: 20 mg/mL (46.68 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2 mg/mL (4.67 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 20.0 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 mg/mL (4.67 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 20.0 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 mg/mL (4.67 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 20.0 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	2.3338 mL	11.6692 mL	23.3383 mL
	5 mM	0.4668 mL	2.3338 mL	4.6677 mL
	10 mM	0.2334 mL	1.1669 mL	2.3338 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.