LEI-401 is a first-in-class, selective, BBB (blood-brain barrier)-passing NAPE-PLD (N-acylphosphoethanolamine phospholipase D) inhibitor (antagonist) with IC50 of 27 nM. Modulation of emotional behavior in mice by LEI-401.

Physicochemical Properties

Molecular Formula	C20H15BRN4O
Molecular Weight	407.263303041458
Exact Mass	421.247
CAS #	2393840-15-6
PubChem CID	145998143
Appearance	White to off-white solid powder
LogP	3.2
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	6
Heavy Atom Count	31
Complexity	609
Defined Atom Stereocenter Count	2
SMILES	BrC1C=CC=CC=1N1C(C)=C(C(NC2C=CC3C=CC=CC=3N=2)=O)C=N1
InChi Key	GFHJYPQZBBHOCC-UXHICEINSA-N
InChi Code	InChI=1S/C24H31N5O2/c30-20-10-12-28(16-20)22-13-21(23(31)25-14-17-8-9-17)26-24(27-22)29-11-4-7-19(15-29)18-5-2-1-3-6-18/h1-3,5-6,13,17,19-20,30H,4,7-12,14-16H2,(H,25,31)/t19-,20+/m1/s1
Chemical Name	N-(cyclopropylmethyl)-6-[(3S)-3-hydroxypyrrolidin-1-yl]-2-[(3S)-3-phenylpiperidin-1-yl]pyrimidine-4-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 neural cells, LEI-401 decreases a variety of NAEs, including anandamide, in a way that is dependent on NAPE-PLD. With an IC50 of 0.86 μM, LEI-401 (0.04 – 20 μM; 30 min) dose-dependently decreased NAPE-PLD staining in hNAPE-transfected HEK293T cells. In Neuro-2a cells, LEI-401 lowers NAE levels, but not in NAPE-PLD KO cells [1].
ln Vivo	In mice, LEI-401 (30 mg/kg; i.p.) decreases fear extinction[1]. Moreover, HPA axis signaling is activated by LEI-401[1]. t1/2, Cmax, tmax, AUClast, and F values of 2.5 hours, 1370 ng/mL, 2 hours, 6760 h*ng/mL, and 25%, respectively, were observed with LEI-401 (10 mg/kg; po) treatment[1]. Cmax, tmax, AUClast, and F values for LEI-401 (30 mg/kg; ip) dosing were 10,300 ng/mL, 1 hour, 38,600 h*ng/mL, and 48%, respectively[1].
Animal Protocol	Animal/Disease Models: Male 7–12weeks old C57BL/6J mice[1] Doses: 30 mg/kg Route of Administration: Ip Experimental Results: Produced a significant increase in freezing as compared to vehicle. Animal/Disease Models: C57BL/6J mice[1] Doses: 10 mg/kg Route of Administration: Po (pharmacokinetic/PK Analysis) Experimental Results: The t1/2, Cmax, tmax, AUClast, and F values were 2.5 hrs (hours), 1370 ng/mL, 2 hrs (hours), 6760 h*ng/mL, and 25 %, respectively.
References	[1]. Discovery of a NAPE-PLD inhibitor that modulates emotional behavior in mice. Nat Chem Biol. 2020;16(6):667-675.

Solubility Data

Solubility (In Vitro)

DMSO: 100 mg/mL (237.23 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.93 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	2.4554 mL	12.2772 mL	24.5543 mL
	5 mM	0.4911 mL	2.4554 mL	4.9109 mL
	10 mM	0.2455 mL	1.2277 mL	2.4554 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.