Lorlatinib 是一种具有口服活性，选择性，脑渗透性和 ATP 竞争性的ROS1/ALK抑制剂，具有抗癌活性。它对 ROS1、野生型 ALK 和 ALKL1196M的Ki值分别为<0.025 nM、<0.07 nM 和 0.7 nM。

PF-06463922 is an orally available, ATP-competitive inhibitor of the receptor tyrosine kinases, anaplastic lymphoma kinase (ALK) and C-ros oncogene 1 (Ros1), with potential antineoplastic activity.

在表达人CD74-ROS1 和 Fig-ROS1的NIH3T3异种移植模型中,PF-06463922通过抑制ROS1磷酸化和下游信号分子,以及对周期蛋白D1的抑制发挥作用,能够抑制细胞增殖.在负荷肿瘤移植物,过表达EML4-ALK,EML4-ALK-L1196M,EML4-ALK-G1269A,EML4-ALK-G1202R 或NPM-ALK的小鼠体内,PF-06463922表现出明显的抗肿瘤活性.

在含有SLC34A2-ROS1 融合物的HCC78人NSCLC细胞和表达人CD74-ROS1的BaF3-CD74-ROS1细胞中,PF-06463922能够抑制细胞增殖,同时诱导细胞凋亡。在含有非突变型ALK或突变型ALK融合物的NSCLC 细胞中, PF-06463922能够抑制细胞增殖,同时诱导细胞凋亡。对ALK和大量ALK临床突变型（IC50=0.2 -77 nM）, PF-06463922表现出明显的细胞活性。

Recombinant human wild-type and mutant ALK kinase domain proteins (amino acids 1093–1411) are produced in-house using baculoviral expression, preactivated via autophosphorylation with MgATP, and assayed for kinase activity using a microfluidic mobility shift assay. The reactions contained 1.3 nM wild-type ALK or 0.5 nM mutant ALK (appropriate to produce 15-20% phosphorylation of peptide substrate after 1 h of reaction), 3 μM 5-FAM-KKSRGDYMTMQIG-CONH2), 5 mM MgCl2, and the Km level of ATP in 25 mM Hepes, pH 7.1. The inhibitors are shown to be ATP-competitive from kinetic and crystallographic studies. The Ki values are calculated by fitting the conversion (%) to a competitive inhibition equation. ROS1 enzyme is assayed 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.

Cells are seeded in 96-well plates in growth medium containing 10% FBS and are cultured overnight at 37°C. The following day, serial dilutions of PF-06463922 or appropriate controls are added to the designated wells, and cells are incubated at 37°C for 72 h. A CellTiter-Glo assay is performed to determine the relative cell numbers. IC50 values are calculated by concentration-response curve fitting using a four-parameter analytical method.

1454846-35-5

C21H19FN6O2

406.41

PF-06463922;PF-6463922;劳拉替尼;Loratinib

DMSO:40.6 mg/mL (100 mM)
Ethanol:40.6 mg/mL (100 mM)
Powder: -20°C for 3 years
In solvent: -80°C for 2 years