1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemELorlatinibCat. No.: HY-12215CAS No.: 1454846-35-5Synonyms: PF-06463922分式: CHFNO分量: 406.41作靶點: ALK作通路: Protein Tyrosine Kinase/RTK儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實驗 DMSO : 28 mg/mL (68

2、.90 mM)H2O : 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (6.15 mM); Clear solution2. 請依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemESolubility: 2.5 mg/mL (6.15 mM); Suspended solution; Need ultrasonic3. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubi

3、lity: 2.5 mg/mL (6.15 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 Lorlatinib (PF-064639220種有效的，ALK/ROS1 雙抑制劑，抑制 ROS1，野ALK 和 突變ALK (L1196M)的 Ki 分別為0.02 nM，0.07 nM 和 0.7 nM。IC50 & Target Ki: 0.02 nM (ROS1), 0.07 nM (ALK WT), 0.7 nM (ALK L1196M)體外研究 Lorlatinib demonstrates significant cell activity again

4、st ALK and a large set of ALK clinical mutations withIC50 ranging from 0.2 nM-77 nM 1. Lorlatinib significantly inhibits cell proliferation and induces cellapoptosis in the HCC78 human NSCLC cells harboring SLC34A2-ROS1 fusions and the BaF3-CD74-ROS1cells expressing human CD74-ROS1 2. Lorlatinib als

5、o shows potent growth inhibitory activity and inducesapoptosis in the NSCLC cells harboring either non-mutant ALK or mutant ALK fusions 3.體內(nèi)研究 In rats, Lorlatinib displays low plasma clearance, a moderate volume of distribution, a reasonable half-life,low propensity for p-glycoprotein 1-mediated eff

6、lux and a bioavailability of 100% 1. In vivo, Lorlatinib showscytoreductive antitumor efficacy in the NIH3T3 xenograft models expressing human CD74-ROS1 and Fig-ROS1 via inhibition in ROS1 phosphorylation and the downstream signaling molecules, as well as inhibitionof the cell cycle protein Cyclin D

7、1 in tumors 2. In vivo, Lorlatinib also demonstrates marked antitumoractivity in mice bearing tumor xenografts expressing EML4-ALK, EML4-ALK-L1196M, EML4-ALK-G1269A,EML4-ALK-G1202R or NPM-ALK 3.PROTOCOLKinase Assay 1 Recombinant human wild-type and mutant ALK kinase domain proteins (amino acids 1093

8、1411) areproduced in-house using baculoviral expression, preactivated via autophosphorylation with MgATP, andassayed for kinase activity using a microfluidic mobility shift assay. The reactions contained 1.3 nM wild-typeALK or 0.5 nM mutant ALK (appropriate to produce 15-20% phosphorylation of pepti

9、de substrate after 1 h ofreaction), 3 M 5-FAM-KKSRGDYMTMQIG-CONH2), 5 mM MgCl2, and the Km level of ATP in 25 mMHepes, pH 7.1. The inhibitors are shown to be ATP-competitive from kinetic and crystallographic studies. TheKi values are calculated by fitting the conversion (%) to a competitive inhibiti

10、on equation. ROS1 enzyme isassayed as described above for ALK, except using 0.25 nM recombinant human ROS1 catalytic domain(amino acids 1883-2347). Kinase inhibitor selectivity is evaluated using a 206-kinase panel.MCE has not independently confirmed the accuracy of these methods. They are for refer

11、ence only.Cell Assay 2 Cells are seeded in 96-well plates in growth medium containing 10% FBS and are cultured overnight at 37C.The following day, serial dilutions of Lorlatinib or appropriate controls are added to the designated wells, andcells are incubated at 37C for 72 h. A CellTiter-Glo assay i

12、s performed to determine the relative cellnumbers. IC50 values are calculated by concentration-response curve fitting using a four-parameter2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEanalytical method.MCE has not independently confirmed the accuracy of these methods. They are for reference onl

13、y.Animal De novoGBM tumorigenesis is initiated in LSL-FIG-ROS1;Cdkn2a/;LSL-Luc mice through intracranialAdministration 2 stereotactic injections of Adeno-Cre as described previously. Tumor development is monitored using BLI asdescribed below. Once tumors reach a given size (107 p-1s-1cm-2sr-1), anim

14、als are randomLy enrolledinto vehicle control or 3-, 7-, or 14-d treatment with the indicated doses of Lorlatinib. Drug is administeredthrough s.c. implanted Alzet osmotic pumps. After treatment, mice are killed, GBM tumors aremicrodissected, and tissues are flash-frozen in liquid N2. The remaining

15、brains are processed for histology.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻 Nat Commun. 2017 Oct 30;8(1):1197. J Anal Methods Chem. 2019 Mar. Harvard Medical School LINCS LIBRARYSee more customer validations on HYPERLINK / www.MedChem

16、EREFERENCES1. Johnson TW, et al. Discovery of (10R)-7-amino-12-fluoro-2,10,16-trimethyl-15-oxo-10,15,16,17-tetrahydro-2H-8,4-(metheno)pyrazolo4,3-h2,5,11-benzoxadiazacyclotetradecine-3-carbonitrile (PF-06463922), a macrocyclic inhibitor of anaplasticlymphoma kinas2. Zou HY, et al. PF-06463922 is a p

17、otent and selective next-generation ROS1/ALK inhibitor capable of blocking crizotinib-resistant ROS1mutations. Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3493-83. Zou HY, et al. PF-06463922 is a potent and selective next-generation ROS1/ALK inhibitor capable of blocking crizotinib-resistant ROS1mutation