ML401 is a potent chemical probe that selectively antagonizes EBI2 (known also as GPR183) with IC50 of 1.03 nM. ML401 showed activity in a chemotaxis assay (IC50=6.24 nM). ML401 displays good stability and is non-cytotoxic.

Physicochemical Properties

Molecular Formula	C20H20BRCLN2O
Molecular Weight	419.742603302002
Exact Mass	418.04
Elemental Analysis	C, 57.23; H, 4.80; Br, 19.04; Cl, 8.45; N, 6.67; O, 3.81
CAS #	1597489-14-9
PubChem CID	73169083
Appearance	Off-white to light yellow solid powder
LogP	4.4
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	2
Rotatable Bond Count	4
Heavy Atom Count	25
Complexity	449
Defined Atom Stereocenter Count	0
SMILES	C1CN(CCN1CC2=CC=C(C=C2)Cl)C(=O)/C=C/C3=CC=C(C=C3)Br
InChi Key	NXTNVQJKGHAGAX-BJMVGYQFSA-N
InChi Code	InChI=1S/C20H20BrClN2O/c21-18-6-1-16(2-7-18)5-10-20(25)24-13-11-23(12-14-24)15-17-3-8-19(22)9-4-17/h1-10H,11-15H2/b10-5+
Chemical Name	(E)-3-(4-bromophenyl)-1-[4-[(4-chlorophenyl)methyl]piperazin-1-yl]prop-2-en-1-one
Synonyms	ML 401; ML-401; ML401
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	EBI2 ( IC50 = 1.03 nM )
ln Vitro	ML401 exhibits no toxicity (>50 μM), and it is non-cytotoxic in MTT in both LnCap and IMR-32 cells (>50 μM) in immortalized Fa2-N4 human hepatocytes[1].
ln Vivo	ML401 exhibits extremely good stability in mouse and human plasma[1].
References	[1]. Functional Antagonists of EBI-2. Probe Reports from the NIH Molecular Libraries Program [Internet].

Solubility Data

Solubility (In Vitro)

DMSO: 10.4~42 mg/mL (24.8~100.1 mM) Ethanol: ~4 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 1.04 mg/mL (2.48 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 10.4 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: ≥ 1.04 mg/mL (2.48 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 10.4 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	2.3824 mL	11.9121 mL	23.8243 mL
	5 mM	0.4765 mL	2.3824 mL	4.7649 mL
	10 mM	0.2382 mL	1.1912 mL	2.3824 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.