Walrycin B is a novel antibacterial compound specifically targeting the essential WalR response regulator. IC50 values: 3.13 ug/ml (MIC for S. aureus N315) and 0.39 ug/ml (MIC for B. subtilis 168). With a strong minimum inhibitory concentration (MIC) for both B. subtilis and S. aureus, toxoflavin—a phytotoxin derived from Burkholderia glumae—has been demonstrated to have an unclear mode of action. Antibacterial Walrycin B is an analog of this toxoflavin.The substance may also have bactericidal effects through interaction with WalR. A novel class of highly effective small molecules known as walrycins targets the RR WalR and inhibits this crucial signal transduction pathway, thus killing bacterial cells. These reagents not only hold therapeutic potential but will also serve as valuable tools for future research on the WalK/WalR TCS. Both B. subtilis and S. aureus experience cell death as a result of Walrycin B's targeting of WalR.

Physicochemical Properties

Molecular Formula	C14H10F3N5O2
Molecular Weight	337.2622
Exact Mass	337.078
Elemental Analysis	C, 49.86; H, 2.99; F, 16.90; N, 20.77; O, 9.49
CAS #	878419-78-4
Related CAS #	878419-78-4;
PubChem CID	3164070
Appearance	Yellow to orange solid powder
Density	1.6±0.1 g/cm3
Boiling Point	383.1±52.0 °C at 760 mmHg
Flash Point	185.5±30.7 °C
Vapour Pressure	0.0±0.9 mmHg at 25°C
Index of Refraction	1.645
LogP	-1.24
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	1
Heavy Atom Count	24
Complexity	674
Defined Atom Stereocenter Count	0
SMILES	O=C1N(C)C(=O)C2=NC(C3C=CC(C(F)(F)F)=CC=3)=NN(C2=N1)C
InChi Key	XRVMPTWWKLKLPB-UHFFFAOYSA-N
InChi Code	InChI=1S/C14H10F3N5O2/c1-21-12(23)9-11(19-13(21)24)22(2)20-10(18-9)7-3-5-8(6-4-7)14(15,16)17/h3-6H,1-2H3
Chemical Name	1,6-dimethyl-3-(4-(trifluoromethyl)phenyl)pyrimido[5,4-e][1,2,4]triazine-5,7(1H,6H)-dione
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	bacterial WalR response regulator
References	[1]. Novel antibacterial compounds specifically targeting the essential WalR response regulator. J Antibiot (Tokyo). 2010 Mar;63(3):127-34. [2]. Fuchs BB, Rajamuthiah R, Jayamani E, Vlahovska PM, Ausubel FM, Mylonakis E. NH125 kills methicillin-resistant Staphylococcus aureus persisters by lipid bilayer disruption. Future Med Chem. 2016;8(3):257-69.

Solubility Data

Solubility (In Vitro)

DMSO : 11.5~40 mg/mL (34.10~118.6 mM) Ethanol : ~2 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: 2.5 mg/mL (7.41 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. 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 (7.41 mM) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. 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	2.9651 mL	14.8254 mL	29.6507 mL
	5 mM	0.5930 mL	2.9651 mL	5.9301 mL
	10 mM	0.2965 mL	1.4825 mL	2.9651 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.