XMU-MP-3 is a potent non-covalent BTK inhibitor (antagonist) with IC50s of 10.7 nM and 17.0 nM for BTK WT and BTK C481S mutant in the presence of 10 μM ATP, respectively. XMU-MP-3 also causes apoptosis.

Physicochemical Properties

Molecular Formula	C27H27F3N8O
Molecular Weight	536.551494836807
Exact Mass	536.225
CAS #	2031152-08-4
PubChem CID	122648639
Appearance	Light yellow to brown solid powder
LogP	5.2
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	10
Rotatable Bond Count	5
Heavy Atom Count	39
Complexity	848
Defined Atom Stereocenter Count	0
InChi Key	MCEWMWAJAOMTBE-UHFFFAOYSA-N
InChi Code	InChI=1S/C27H27F3N8O/c1-16-8-9-21(32-25(39)18-6-5-7-20(11-18)27(28,29)30)12-22(16)38-14-19-13-31-26(34-24(19)36(3)15-38)33-23-10-17(2)35-37(23)4/h5-13H,14-15H2,1-4H3,(H,32,39)(H,31,33,34)
Chemical Name	N-[3-[2-[(2,5-dimethylpyrazol-3-yl)amino]-8-methyl-5,7-dihydropyrimido[4,5-d]pyrimidin-6-yl]-4-methylphenyl]-3-(trifluoromethyl)benzamide
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	IC50: 10.7 nM (BTK WT), 17.0 nM (BTK C481S), Apoptosis[1]
ln Vitro	XMU-MP-3 suppresses the growth of BTK-transformed Ba/F3 cells with a 48-hour half-life of 0.001–10,000 nM [1]. Ramos, JeKo-1, and NALM-6 are all inhibited by XMU-MP-3 (1–1000 nM), with IC50 values of 326.6 nM, 685.6 nM, and 1065 nM, respectively[1]. -cells Ba/F3 [1]. Ba/F3 cells from BTK (C481S) undergo apoptosis when exposed to XMU-MP-3 at 5000 nM[1]. In Ba/F3 cells transfected with BTK, there is dose-dependent auto- and transphosphorylation of BTK at the Y223 and Y551 sites[1].
ln Vivo	In mouse xenograft models, XMU-MP-3 (25 and 50 mg/kg) significantly inhibits tumor growth[1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: BTK-transformed and parental Ba/F3 cells Tested Concentrations: 0.001, 0.01, 0.1, 1, 10, 100, 1000, 10000 nM Incubation Duration: 48 hrs (hours) Experimental Results: Inhibited BTK-transformed Ba/F3 cell proliferation with an IC50 of 11.4 nM, while it demonstrated negligible anti-proliferative effects on parental wild-type Ba/F3 cells (IC50 >10000 nM). Western Blot Analysis[1] Cell Types: BTK-transformed Ba/F3 cells Tested Concentrations: 10, 50, 100, 500, 1000 nM Incubation Duration: 4 hrs (hours) Experimental Results: The phosphorylation levels of BTK Y223 and Y551 were Dramatically decreased at concentrations as low as 100 nM, and completely suppressed at the concentration of 1000 nM.
Animal Protocol	Animal/Disease Models: Nu/nu BALB/ c mice (4-6 weeks of age) bearing BTK-transformed Ba/F3 and Ramos xenograft models[1] Doses: 25 and 50 mg/kg Route of Administration: Treated by tail vein injection; the injection volume was 0.1 mL per 10 g; daily for 14 days Experimental Results: Dramatically diminished the tumor size without affecting animal weights.
References	[1]. A Non-Covalent Inhibitor XMU-MP-3 Overrides Ibrutinib-Resistant Btk C481S Mutation in B-cell Malignancies. Br J Pharmacol. 2019 Dec;176(23):4491-4509.

Solubility Data

Solubility (In Vitro)

DMSO : 100 mg/mL (186.38 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.66 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 (4.66 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	1.8638 mL	9.3188 mL	18.6376 mL
	5 mM	0.3728 mL	1.8638 mL	3.7275 mL
	10 mM	0.1864 mL	0.9319 mL	1.8638 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.