RN486 (RN-486) is a novel, potent, reversible and selective BTK (Bruton's tyrosine kinase) inhibitor with potential anti-inflammatory activity. It inhibits BTK with an IC50 of 4 nM. RN486 exhibits strong in vivo anti-inflammatory and bone-protective effects in mouse CIA and rat adjuvant-induced arthritis (AIA) models. In the AIA model, RN486 inhibits both joint and systemic inflammation either alone or in combination with methotrexate, reducing both paw swelling and inflammatory markers in the blood.

Physicochemical Properties

Molecular Formula	C35H35FN6O3
Molecular Weight	606.69
Exact Mass	606.275
CAS #	1242156-23-5
Related CAS #	1242156-23-5
PubChem CID	46908026
Appearance	Off-white to light yellow solid powder
Density	1.4±0.1 g/cm3
Boiling Point	868.6±65.0 °C at 760 mmHg
Flash Point	479.1±34.3 °C
Vapour Pressure	0.0±0.3 mmHg at 25°C
Index of Refraction	1.696
LogP	2.95
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	7
Heavy Atom Count	45
Complexity	1210
Defined Atom Stereocenter Count	0
InChi Key	ZTUJNJAKTLHBEX-UHFFFAOYSA-N
InChi Code	InChI=1S/C35H35FN6O3/c1-39-12-14-41(15-13-39)26-8-9-32(37-19-26)38-30-18-25(20-40(2)34(30)44)27-4-3-5-31(28(27)21-43)42-11-10-23-16-24(22-6-7-22)17-29(36)33(23)35(42)45/h3-5,8-11,16-20,22,43H,6-7,12-15,21H2,1-2H3,(H,37,38)
Chemical Name	6-cyclopropyl-8-fluoro-2-(2-hydroxymethyl-3-{1-methyl-5-[5-(4-methyl-piperazin-1-yl)-pyridin-2-ylamino]-6-oxo-1,6-dihydro-pyridin-3-yl}-phenyl)-2H-isoquinolin-1-one
Synonyms	RN-486; RN 486; RN486;
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

ln Vitro	Fcγ receptor engagement-mediated tumor necrosis factor α production in monocytes (IC50 = 7 nM), Fcε receptor cross-linking-induced degranulation in mast cells (IC50 = 2.9 nM), and B cell antigen receptor-induced expression of an activation marker, CD69, in B cells in whole blood (IC50 = 21 nM) are all blocked by RN486[1]. Convulxin stimulation led to increased production of pro-inflammatory cytokines, IL-6 and IL-8, in a co-culture system including human primary synovial FLS and activated human platelets. This effect is dose-dependently inhibited by RN486[2].
ln Vivo	In rat adjuvant-induced arthritis (AIA) and mouse CIA models, RN486 has strong anti-inflammatory and bone-protective properties. In the AIA model, paw edema and inflammatory markers in the blood are decreased by RN486 (1–30 mg/kg), which also reduces joint and systemic inflammation[1].
Animal Protocol	N/A Rodent models
References	[1]. RN486, a selective Bruton's tyrosine kinase inhibitor, abrogates immune hypersensitivity responses and arthritis in rodents. J Pharmacol Exp Ther. 2012 Apr;341(1):90-103. [2]. Bruton's Tyrosine Kinase mediates platelet receptor-induced generation of microparticles: a potential mechanism for amplification of inflammatory responses in rheumatoid arthritis synovial joints. Immunol Lett. 2013 Feb;150(1-2):97-104. [3]. Suppression of glomerulonephritis in lupus-prone NZB × NZW mice by RN486, a selective inhibitor of Bruton's tyrosine kinase. Arthritis Rheum. 2013 Sep;65(9):2380-91.

Solubility Data

Solubility (In Vitro)	DMSO: 62 mg/mL (102.2 mM) Water:<1 mg/mL Ethanol:<1 mg/mL
Solubility (In Vivo)	5% DMSO+30% PEG 300+10% Tween 80+ddH2O: 5mg/mL  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.6483 mL	8.2414 mL	16.4829 mL
	5 mM	0.3297 mL	1.6483 mL	3.2966 mL
	10 mM	0.1648 mL	0.8241 mL	1.6483 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.