MRL-494 HCl is an antibacterial agent, an inhibitor (blocker/antagonist) of BamA, and has resistance to efflux and outer membrane permeability. MRL-494 HCl inhibits Gram-positive (Gram+) bacteria (Staphylococcus aureus COL with a MIC of 12.5 μM) and Gram-negative (Gram+) bacteria (E. coli JCM158 with a MIC of 25 μM).

Physicochemical Properties

Molecular Formula	C26H36CLFN16O2
Molecular Weight	659.12
Exact Mass	658.287
CAS #	2699937-04-5
Related CAS #	MRL-494;2434898-43-6
PubChem CID	154925628
Appearance	Typically exists as solid at room temperature
Hydrogen Bond Donor Count	8
Hydrogen Bond Acceptor Count	12
Rotatable Bond Count	12
Heavy Atom Count	46
Complexity	1050
Defined Atom Stereocenter Count	0
InChi Key	OUQBLPVSJWIYJV-UHFFFAOYSA-N
InChi Code	InChI=1S/C26H35FN16O2.ClH/c27-15-5-3-14(4-6-15)21-40-42-43(41-21)12-20(45)32-16-7-9-17(10-8-16)33-24-37-25(39-26(38-24)36-23(30)31)34-18(13-1-2-13)11-19(44)35-22(28)29;/h3-6,13,16-18H,1-2,7-12H2,(H,32,45)(H4,28,29,35,44)(H6,30,31,33,34,36,37,38,39);1H
Chemical Name	3-cyclopropyl-N-(diaminomethylidene)-3-[[4-(diaminomethylideneamino)-6-[[4-[[2-[5-(4-fluorophenyl)tetrazol-2-yl]acetyl]amino]cyclohexyl]amino]-1,3,5-triazin-2-yl]amino]propanamide;hydrochloride
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 Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage.(2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.
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	β-barrel assembly machine A (BamA)[1]
ln Vitro	MRL-494 causes the cytoplasmic membrane to rupture fatally. By focusing on BamA, MRL-494 prevents the synthesis of OM proteins (OMPs) from outside the outer membrane (OM). Strong anti-microbial effects against both Gram-positive and Gram-negative bacteria are demonstrated by MRL-494. The MRL-494 MIC values against E. Coli (WT), E. Coli (ΔtolC), E. K. Coli (\tolC envA101). pneumonia, A. baumannii (WT), A. Baumannii (ΔlpxC), Pseudomonas aeruginosa (efflux deficient), Pseudomonas aeruginosa (WT), Methicillin-resistant Staphylococcus aureus, and Bacillus subtilis rpoB18 have respective concentrations of 25 μM, 25 μM, 25 μM, 100 μM, 200 μM, 100 μM, 12.5 μM, and 25 μM[1].
References	[1]. Hart EM, A small-molecule inhibitor of BamA impervious to efflux and the outer membrane permeability barrier. Proc Natl Acad Sci U S A. 2019 Oct 22;116(43):21748-21757.

Solubility Data

Solubility (In Vitro)	H2O : 110 mg/mL (166.89 mM) DMSO : 100 mg/mL (151.72 mM)
Solubility (In Vivo)	Solubility in Formulation 1: 25 mg/mL (37.93 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with heating and sonication.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.5172 mL	7.5859 mL	15.1717 mL
	5 mM	0.3034 mL	1.5172 mL	3.0343 mL
	10 mM	0.1517 mL	0.7586 mL	1.5172 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.