GSK3494245 (DDD01305143) is a potent, orally bioactive, selective inhibitor of chymotrypsin-like activity that binds to the parasite proteasome at a site sandwiched between the β4 and β5 subunits (for WT L. donovani proteasome IC50=0.16μM). GSK3494245 moderately inhibits the chymotrypsin-like activity of the human proteasome (IC50= 26S=13 µM; enriched THP-1 extract IC50=40 µM). GSK3494245 has good biosafety properties.

Physicochemical Properties

Molecular Formula	C21H23FN6O2
Molecular Weight	410.4447
Exact Mass	410.186
CAS #	2080410-41-7
PubChem CID	126571915
Appearance	White to yellow solid powder
LogP	2.6
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	3
Heavy Atom Count	30
Complexity	600
Defined Atom Stereocenter Count	0
SMILES	FC1C([H])=C([H])C(=C([H])C=1C1C([H])=C([H])N2C(N=1)=NC([H])=C2N1C([H])([H])C([H])([H])OC([H])([H])C1([H])[H])N([H])C(N1C([H])([H])C([H])([H])C([H])([H])C1([H])[H])=O
InChi Key	SAJUCKZZYFFICP-UHFFFAOYSA-N
InChi Code	InChI=1S/C21H23FN6O2/c22-17-4-3-15(24-21(29)27-6-1-2-7-27)13-16(17)18-5-8-28-19(14-23-20(28)25-18)26-9-11-30-12-10-26/h3-5,8,13-14H,1-2,6-7,9-12H2,(H,24,29)
Chemical Name	N-[4-fluoro-3-(3-morpholin-4-ylimidazo[1,2-a]pyrimidin-7-yl)phenyl]pyrrolidine-1-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

ln Vitro	In the L. donovani intramacrophage experiment, where the amastigotes are cultivated in differentiated THP-1 cells, GSK3494245 exhibits an EC50 value of 5.7 μM. Good selectivity against mammalian cell growth inhibition (THP-1 cells; EC50 > 50 μM) is demonstrated by GSK3494245[1]. The pEC50 values for GSK3494245 (DDD01305143) against axenic amastigote and ld InMac are 6.5 and 5.8, respectively. The Ld InMac test for intramacrofaging is performed using L. donovani amastigote in THP-1 cells[2].
ln Vivo	In infected mice (L. donovani, LV9), GSK3494245 (25 mg/kg; taken twice daily for 10 days in a row) results in a >95% reduction in the parasite load[1].
References	[1]. Preclinical candidate for the treatment of visceral leishmaniasis that acts through proteasome inhibition. Proc Natl Acad Sci U S A. 2019;116(19):9318-9323. [2]. Identification of GSK3186899/DDD853651 as a Preclinical Development Candidate for the Treatment of Visceral Leishmaniasis. J Med Chem. 2019;62(3):1180-1202.

Solubility Data

Solubility (In Vitro)

DMSO : 100 mg/mL (243.64 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.09 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 (6.09 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.4364 mL	12.1820 mL	24.3641 mL
	5 mM	0.4873 mL	2.4364 mL	4.8728 mL
	10 mM	0.2436 mL	1.2182 mL	2.4364 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.