BMS-935177 is a potent, selective, reversible, and 2nd generation inhibitor of Bruton’s Tyrosine Kinase (BTK) with an IC50 of 3 nM. It outperforms other kinases, including those of the Tec family (TEC, BMX, ITK, and TXK, among others), in terms of kinase selectivity (5- and 67-fold selectivity) and activity against BTK. Clinical applications for BMS-935177 include the treatment of autoimmune diseases. BMS-935177 inhibited CD69 surface expression in peripheral B cells stimulated with anti-IgM and anti-IgG, and it also inhibited calcium flux in human Ramos B cells with an IC50 of 27 nM. BMS-935177 was chosen to proceed into clinical development due to its notable in vitro potency, strong in vivo activity, good pharmacokinetic profile, and tolerable safety profile.

Physicochemical Properties

Molecular Formula	C31H26N4O3
Molecular Weight	502.56
Exact Mass	502.2
Elemental Analysis	C, 74.09; H, 5.21; N, 11.15; O, 9.55
CAS #	1231889-53-4
Related CAS #	1231889-53-4
PubChem CID	70647728
Appearance	Light yellow to yellow solid powder
Density	1.4±0.1 g/cm3
Boiling Point	805.4±75.0 °C at 760 mmHg
Flash Point	440.9±37.1 °C
Vapour Pressure	0.0±3.0 mmHg at 25°C
Index of Refraction	1.707
LogP	3.98
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	4
Heavy Atom Count	38
Complexity	951
Defined Atom Stereocenter Count	0
SMILES	OC(C)(C)C1C=CC2=C(C=1)NC1C(C(N)=O)=CC=C(C3C=CC=C(C=3C)N3C=NC4C=CC=CC=4C3=O)C2=1
InChi Key	TVJRDCQUZMGBAB-UHFFFAOYSA-N
InChi Code	InChI=1S/C31H26N4O3/c1-17-19(8-6-10-26(17)35-16-33-24-9-5-4-7-22(24)30(35)37)20-13-14-23(29(32)36)28-27(20)21-12-11-18(31(2,3)38)15-25(21)34-28/h4-16,34,38H,1-3H3,(H2,32,36)
Chemical Name	7-(2-hydroxypropan-2-yl)-4-[2-methyl-3-(4-oxoquinazolin-3-yl)phenyl]-9H-carbazole-1-carboxamide
Synonyms	BMS-935177; BMS935177; BMS 935177
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	BTK (IC50 = 2.8 nM); TEC (IC50 = 13 nM); BLK (IC50 = 20 nM); BMX (IC50 = 24 nM); TrkA (IC50 = 30 nM)
ln Vitro	BMS-935177 exhibits selectivity over the SRC family of kinases by more than 50 times, including 110% selectivity over SRC. The remaining kinases, TRKA, HER4, TRKB, and RET, were inhibited with a potency less than 150 nM (50-fold selectivity). When peripheral B cells are stimulated with anti-IgM and anti-IgG, it suppresses CD69 surface expression and calcium flux in human Ramos B cells (IC50 = 27 nM). But when B cells are stimulated via the CD40 receptor using CD40 ligand, BMS-935177 has no effect on the expression of CD69 on their surface. With an IC50 value of 14 nM, BMS-935177 effectively inhibited TNFα production against IgG-containing immune complex-driven low affinity activating Fcγ receptor (FcγRIIa and FcγRIII) end points in peripheral blood mononuclear cells (PBMCs)[1].
ln Vivo	For BMS-935177, plasma protein binding is high across all species, with less than 1% free for humans. In spite of its low aqueous solubility, it has good oral bioavailability in all preclinical species, from both suspension and solution dosing. BMS-935177 has an oral bioavailability of 84% to 100% when given as a solution dose in rat, mouse, dog, and cynomolgus monkey studies. Single intravenous (IV) infusion studies show low clearance. The half-lives of BMS-935177 in mice and rats administered intravenously at a dose of 2 mg/kg are 4 and 5.1 hours, respectively[1].
Enzyme Assay	BMS-935177 is tested at 11 different concentrations after being dissolved at 10 mM in DMSO. BMS-935177, 1 nM of human recombinant BTK, 1.5 μM of fluoresceinated peptide, 20 μM (Km app) of ATP, and 20 mM HEPES, pH 7.4, 10 mM MgCl2, 0.015% Brij 35 surfactant, and 4 mM DTT in 1.6% DMSO) are added to V-bottom 384-well plates, with a final volume of 30 μL of assay buffer. 45 μL of 35 mM EDTA is added to each sample to stop the reaction after it has been incubated for 60 minutes at room temperature. By electrophoretic separation of the fluorescent substrate and phosphorylated product (excitation, 488 nm; emission, 530 nm), the reaction mixture is examined.
Animal Protocol	Rats: In the PK studies, male Sprague-Dawley rats weighing 255-298 g are employed. Rats are given T99.5% 10 mM citrate buffer, pH 4, 0.02% DOSS, Methocel A4M, and BMS-935177 by oral gavage (1, 5, and 20 mg/kg) in order to examine the oral bioavailability of the drug following crystalline microsuspension doses. Following oral dosage, serial blood samples are taken at 0.25, 0.5, 0.75, 1, 2, 4, 6, 8, and 24 hours later. After centrifugation at 4 °C (1500-2000g), plasma samples are kept at -20 °C until analysis. Mice: BMS-935177 (10 and 30 mg/kg) is tested in a mouse anti-collagen antibody-induced arthritis (CAIA) inflammation model to compare its effectiveness with vehicle and dexamethasone (Dex). Four different combinations of monoclonal antimouse type II collagen antibodies—one milligram of each—are injected intraperitoneally (ip) into mice. The vehicle (EtOH:TPGS:PEG300, 5:5:90), BMS-935177 (10 or 30 mg/kg), or dexamethasone (dex, 1 mg/kg) is immediately started as a daily oral dose. The mice receive an intraperitoneal injection of 1.25 mg/kg lipopolysaccharide (LPS) after three days. The mice are then observed three times a week to observe the progression and intensity of paw inflammation.
References	[1]. Small Molecule Reversible Inhibitors of Bruton's Tyrosine Kinase (BTK): Structure-Activity Relationships Leading to the Identification of 7-(2-Hydroxypropan-2-yl)-4-[2-methyl-3-(4-oxo-3,4-dihydroquinazolin-3-yl)phenyl]-9H-carbazole-1-c.

Solubility Data

Solubility (In Vitro)

DMSO: >100 mg/mL Water: < 1 mg/mL Ethanol: >100 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: 3.75 mg/mL (7.46 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 37.5 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: 3.75 mg/mL (7.46 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 37.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.9898 mL	9.9491 mL	19.8981 mL
	5 mM	0.3980 mL	1.9898 mL	3.9796 mL
	10 mM	0.1990 mL	0.9949 mL	1.9898 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.