RN486 是选择性的，具有口服活性的BTK抑制剂 (IC50:4.0 nM;Kd:0.31 nM)，对其他激酶的活性较低。它可用于类风湿关节炎和系统性红斑狼疮的研究。

RN486 is an effective and specific BTK inhibitor (IC50: 4 nM).

在AIA模型中,RN486单独或与甲氨蝶呤联用,可使关节和全身性炎症受到抑制,并使足肿胀和血液中炎症标志物减少.在啮齿动物模型中,RN486的功能活性类似,有效防止I型和III型超敏反应.且在小鼠CIA和大鼠佐剂性关节炎模型中,RN486具有较强的骨保护和抗炎作用.

RN486既可选择性抑制Bruton酪氨酸激酶,又在多种细胞类型的人类细胞试验中表现出功能活性,阻断Fcε受体交联诱导的肥大细胞脱粒(IC50：2.9 nM),Fcγ受体衔接介导的单核细胞中肿瘤坏死因子α的产生(IC50：7.0 nM),以及B细胞抗原受体诱导的活化标志物CD69在全血B细胞中的表达(IC50：21.0 nM)。

Radiometric kinase assay: The enzymatic kinase activity is assessed by measuring the phosphorylation of a synthetic substrate by the purified GST-fusion FGFR3-K650E kinase domain, in the presence of radiolabeled ATP. Enzyme activities are measured by mixing 10 μL of a 3-fold concentrated BGJ398 solution or control with 10 μL of the corresponding substrate mixture (peptidic substrate, ATP and [γ33P]ATP). The reactions are initiated by addition of 10 μL of a 3-fold concentrated solution of the enzyme in assay buffer. The final concentrations of the assay components are as following: 10 ng of GST-FGFR3-K650E, 20 mM Tris-HCl, pH 7.5, 3 mM MnCl2, 3 mM MgCl2, 1 mM DTT, 250 μg/mL PEG 20000, 2 μg/mL poly(EY) 4:1, 1% DMSO and 0.5 μM ATP (γ-[33P]-ATP 0.1 μCi). The assay is carried out according to the filter binding (FB) method in 96-well plates at room temperature for 10 minutes in a final volume of 30 μL including BGJ398. The enzymatic reactions are stopped by the addition of 20 μL of 125 mM EDTA, and the incorporation of 33P into the polypeptidic substrates is quantified as following: 30 μL of the stopped reaction mixture are transferred onto Immobilon-PVDF membranes previously soaked for 5 minutes with methanol, rinsed with water, soaked for 5 min with 0.5% H3PO4, and mounted on vacuum manifold with disconnected vacuum source. After spotting, vacuum is connected, and each well rinsed with 0.5% H3PO4 (200 μL). Free membranes are removed and ished four times on a shaker with 1% H3PO4 and once with ethanol. Membranes are dried and overlaid with addition of 10 μL/well of a scintillation fluid. The plates are eventually sealed and counted in a microplate scintillation counter. IC50 values are calculated by linear regression analysis of the percentage inhibition of the BGJ398.

1242156-23-5

C35H35FN6O3

606.702

DMSO:58 mg/mL (95.6 mM)
Ethanol:<1 mgml
H2O:<1 mgml
Powder: -20°C for 3 years
In solvent: -80°C for 2 years