1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEInfigratinib phosphateCat. No.: HY-13311ACAS No.: 1310746-10-1Synonyms: BGJ-398 phosphate; NVP-BGJ398 (phosphate)分式: CHClNOP分量: 658.47作靶點: FGFR作通路: Protein Tyrosine Kinase/RTK儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 m

2、onths-20C 1 monthBIOLOGICAL ACTIVITY物活性 Infigratinib phosphate (BGJ-398 phosphate)是FGFR抑制劑，對FGFR1，FGFR2和FGFR3的IC50分別為0.9，1.4和1 nM，對FGFR4和VEGFR2的抑制性40倍，對Abl，Fyn，Kit，Lck和Lyn乎活性。IC50 & Target FGFR1 FGFR2 FGFR3 FGFR40.9 nM (IC50) 1.4 nM (IC50) 1 nM (IC50) 60 nM (IC50)體外研究 Infigratinib phosphate inhibits

3、 FGFR1, FGFR2, and FGFR3 with IC50=1 nM, FGFR3K650E with IC50=4.9nM, and FGFR4 with IC50=60 nM. IC50 values for all other kinases are in the M range (FYN, LCK, YES,and ABL, IC50=1.9, 2.5, 1.1, and 2.3 M, respectively) except for VEGFR2, KIT, and LYN, which areinhibited at submicromolar concentration

4、s (IC50=0.18, 0.75, and 0.3 M, respectively). Infigratinib inhibits theproliferation of the FGFR1-, FGFR2-, and FGFR3-dependent BaF3 cells with IC50 values which are in thelow nanomolar range and comparable to those observed for the inhibition of the receptors kinase activity inthe enzymatic assay.

5、For the remaining cells, all IC50 values are greater than 1.5 M except for VEGFR2(IC50 1449 and 938 nM), for which there is at least a 400-fold selectivity versus FGFR1, FGFR2, and FGFR31. Infigratinib (ranging between 1 nM and 10 M) is potent at inhibiting cell growth of FGFR2-mutantendometrial can

6、cer cells 2.體內研究Infigratinib is administered to athymic nude mice implanted subcutaneously with RT112/luc1 tumors: eitheras a 5 mg/kg intravenous bolus in NMP/PEG200 (1:9, v/v) or orally by gavage as a suspension in1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEPEG300/D5W (2:1, v/v) at a 20 mg/kg

7、dose. The relevant pharmacokinetic (PK) parameters indicate that theoral bioavailability of Infigratinib in this study is 32%. After intravenous dosing, Infigratinib shows a rapiddistribution from the vascular compartment into the peripheral tissues, translating into a high volume ofdistribution (26

8、 L/kg). The plasma clearance is high at 3.3 L/h/kg (61% of liver blood flow). The ratio of tumorto plasma after oral dosing based on AUC is determined to be 10 1. Infigratinib (30 mg/kg) significantlyinhibits the growth of FGFR2-mutated endometrial cancer xenograft models 2.PROTOCOLKinase Assay 1 Th

9、e enzymatic kinase activity is assessed by measuring the phosphorylation of a synthetic substrate by thepurified GST-fusion FGFR3-K650E kinase domain, in the presence of radiolabeled ATP. Enzyme activitiesare measured by mixing 10 L of a 3-fold concentrated Infigratinib solution or control with 10 L

10、 of thecorresponding substrate mixture (peptidic substrate, ATP and 33PATP). The reactions are initiated byaddition of 10 L of a 3-fold concentrated solution of the enzyme in assay buffer. The final concentrations ofthe assay components are as following: 10 ng of GST-FGFR3-K650E, 20 mM Tris-HCl, pH

11、7.5, 3 mM MnCl2,3 mM MgCl2, 1 mM DTT, 250 g/mL PEG 20000, 2 g/mL poly(EY) 4:1, 1% DMSO and 0.5 M ATP (-33P-ATP 0.1 Ci). The assay is carried out according to the filter binding (FB) method in 96-well plates at roomtemperature for 10 min in a final volume of 30 L including the components as indicated

12、 above. Theenzymatic reactions are stopped by the addition of 20 L of 125 mM EDTA, and the incorporation of 33P intothe polypeptidic substrates is quantified as following: 30 L of the stopped reaction mixture are transferredonto Immobilon-PVDF membranes previously soaked for 5 min with methanol, rin

13、sed with water, soaked for5 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 L). Free membranes are removed andwashed four times on a shaker with 1% H3PO4 and once with ethanol. Membrane

14、s are dried and overlaidwith addition of 10 L/well of a scintillation fluid. The plates are eventually sealed and counted in a microplatescintillation counter. IC50 values are calculated by linear regression analysis of the percentage inhibition ofNVP-BGJ398 1.MCE has not independently confirmed the

15、 accuracy of these methods. They are for reference only.Cell Assay 1 Murine BaF3 cell lines are cultured in RPMI-1640 media supplemented with 10% FBS, 4.5 g/L glucose, 1.5g/L sodium bicarbonate, and Pen/Strep. Cells are passaged twice weekly. Compound-mediated inhibition ofBaF3 cell proliferation an

16、d viability is assessed using a Luciferase bioluminescent assay. Exponentiallygrowing BaF3 or BaF3 Tel-TK cells are seeded into 384-well plates (4250 cells/well) at 50 L/well using a Fill liquid dispenser in fresh medium. Infigratinib is serially diluted in DMSO and arrayed in a polypropylene384-wel

17、l plate. Then 50 nL of compound are transferred into the plates containing the cells by using thepintool transfer device, and the plates incubated at 37C (5% CO2) for 48 h. Then 25 L of Bright-Glo areadded, and luminescence is quantified using an Analyst-GT. Custom curve-fitting software is used to

18、producea logistic fit of percent cell viability as a function of the logarithm of inhibitor concentration. The IC50 value isdetermined as the concentration of compound needed to reduce cell viability to 50% of a DMSO control 1.MCE has not independently confirmed the accuracy of these methods. They a

19、re for reference only.Animal Mice 1Administration 1 Female HsdNpa: Athymic Nude-nu mice are used. Infigratinib is formulated as a suspension in PEG300/D5W2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemE(2:1, v/v) and administered orally for 12 consecutive days at the doses of 10 and 30 mg/kg/qd. Tu

20、mor andbody weight data are analyzed by ANOVA with post hoc Dunnetts test for comparison of treatment versuscontrol group. The post hoc Tukey test is used for intragroup comparison. Statistical analysis is performedusing GraphPad prism 4.02. As a measure of efficacy, the T/C (%) value is calculated.

21、Rats 1Female nude Rowett rats 6-9 weeks of age are used. Infigratinib is formulated as a solution in acetic acid-acetate buffer pH 4.6/PEG300 (1:1, v/v) and applied daily by gavage to the tumor-bearing rats (n=8) for 20consecutive days at doses of 5, 10, and 15 mg/kg/qd (free base equivalents). The

22、application volume is 5mL/kg. Tumor volumes are measured with calipers and determined according to the formula:lengthwidthheight/6. Antitumor activity is expressed as T/C (%): (mean change of tumor volume oftreated animals/mean change of tumor volume of control animals)100. Regressions (%) are calculated.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產品發(fā)表的科研獻 Cancer Discov. 2018 Mar;8(3):354-369. Sci Transl Med. 2018 Jul 18;10(450). pii: eaaq1093. Ann Rheum Dis. 2016 May;75(5):883-90. J Control Release. 2018 Sep 28;286:254-263.