PF-06447475

製品コードS8202 バッチS820201

印刷

化学情報

Chemical Structure Synonyms N/A Storage
(From the date of receipt)		 3 years -20°C powder
1 years -80°C in solvent
化学式

C17H15N5O
分子量 305.33 CAS No. 1527473-33-1
Solubility (25°C)* 体外 DMSO 61 mg/mL (199.78 mM)
Water Insoluble
Ethanol Insoluble
体内 （毎回新しく調製した物を用意してください）
Homogeneous suspension
CMC-NA
≥5mg/ml Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

溶剤液（一定の濃度）を調合する

生物活性

製品説明 PF-06447475 is a potent, selective, and brain penetrant LRRK2 kinase inhibitor with IC50 of 3 nM.
in vitro In the macrophage cell line Raw264.7, PF-06447475 inhibits endogenous LRRK2 kinase activity with IC50 of <10 nM. [2] In astrocytes, this compound rescues LRRK2 mutation-induced defects in lysosomal morphology and function. [3]
in vivo In G2019S BAC-transgenic mice, PF-06447475 (100 mg/kg, p.o.) inhibits pS935 and pS1292 phosphorylation of LRRK2 with IC50 of 103 nM and 21 nM, respectively. [1] In G2019S-LRRK2 rats, this compound (30 mg/kg, p.o.) blocks α-synuclein-induced dopaminergic neurodegeneration and attenuates neuroinflammation associated with G2019S-LRRK2 expression. [2]

プロトコル(参考用のみ)

キナーゼアッセイ LRRK2 Biochemical Assay
LRRK2 kinase activity is measured using Lantha Screen technology. GST-tagged truncated LRRK2 or mutant G2019S LRRK2 is incubated with fluorescein-labeled peptide substrate LRRKtide in the presence of a dose response of this compound. Upon completion, the assay is stopped and detected with a terbium labeled anti-phospho-ERM antibody. The assay is carried out under the following protocol: 3 μL of a working solution of substrate (233 nM LRRKtide, 117 μM ATP) prepared in assay buffer (50 mM HEEPES, pH 7.5, 3 mM MgCl2, with 2 mM DTT and 0.01% Brij35 added fresh) is added to a low volume Greiner 384-well plate. The compound dose response is prepared by diluting this chemical to a top concentration of 3.16 mM in 100% DMSO and serial diluted by half-log in DMSO 11 times. Aliquots (3.5 μL) of the 100% DMSO dose response are mixed with 46.5 μL of water, and then 1 μL of this mixture is added to the 3 μL of substrate mix in the 384-well plate. The kinase reaction is started with 3 μL of a working solution of LRRK2 enzyme at a concentration of 4 μg/mL. The final reaction concentrations are 100 nM LRRKtide, 50 μM ATP, 1.7 μg/mL LRRK2 enzyme, and a compound dose response with a top dose of 32 μM. The reaction is allowed to progress at room temperature for 2 h or 90 min for the mutant protein and then stopped with the addition of 7 μL of detection buffer (20 mM Tris, pH 7.6, 0.01% NP-40, 0.02% NaN3, 6 mM EDTA with 2 nM terbium labeled anti-phospho-ERM). After an incubation of 1 h at room temperature, the plate is read on an Envision with an excitation wavelength of 340 nm and a reading emission at both 520 and 495 nm. The ratio of the 520 and 495 nm emission is used to analyze the data.
動物実験 動物モデル G2019S BAC-transgenic mice
投薬量 100 mg/kg
投与方法 p.o.

参考

  • https://pubmed.ncbi.nlm.nih.gov/25353650/
  • https://pubmed.ncbi.nlm.nih.gov/26078453/
  • https://pubmed.ncbi.nlm.nih.gov/26251043/

長期の保管のために-20°Cの下で製品を保ってください。

人間や獣医の診断であるか治療的な使用のためにでない。

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