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

Loratinib;PF-6463922;PF-06463922;劳拉替尼;Lorlatinib

[1]Zou HY, et al. Mol Cancer Ther. 2013, 12, A277. [2]Wenlong Li, Rolf W. Sparidans, Yaogeng Wang, Maria C. Lebre, Els Wagenaar, Jos H. Beijnen, and Alfred H. P-glycoprotein (MDR1/ABCB1) restricts brain accumulation and Cytochrome P450-3A (CYP3A) limits oral availability of the novel ALK/ROS1 inhibitor lorlatinib [J]. International journal of cancer. 2018 Oct 15;143(8):2029-2038. [3]Zou HY, et al. Mol Cancer Ther. 2013, 12, C253. [4]Spatari, Claudia, et al. Bioanalytical assay for the quantification of the ALK inhibitor lorlatinib in mouse plasma using liquid chromatography-tandem mass spectrometry. Journal of Chromatography B. 2018 Apr 15;1083:204-208. [5]Sparidans R W, Li W, Schinkel A H, et al. Bioanalytical liquid chromatography-tandem mass spectrometric assay for the quantification of the ALK inhibitors alectinib, brigatinib and lorlatinib in plasma and mouse tissue homogenates[J]. Journal of pharmaceutical and biomedical analysis. 2018 Nov 30;161:136-143. [6]Wenlong Li, Rolf W. Sparidans, Yaogeng Wang, Maria C. Lebre, Jos H. Beijnen, and Alfred H. Schinkel. Oral coadministration of elacridar and ritonavir enhances brain accumulation and oral availability of the novel ALK/ROS1 inhibitor lorlatinib [J]. European Journal of Pharmaceutics and Biopharmaceutics. 2019 Mar;136:120-130. [7]Johnson TW, et al. J Med Chem. 2014, 57(11), 4720-4744.

[1]Li W, Sparidans R W, Wang Y, et al. P-glycoprotein (MDR1/ABCB1) restricts brain accumulation and Cytochrome P450-3A (CYP3A) limits oral availability of the novel ALK/ROS1 inhibitor lorlatinib. International Journal of Cancer. 2018, 143(8): 2029-2038 [2]Li W, Sparidans R W, Wang Y, et al. Oral coadministration of elacridar and ritonavir enhances brain accumulation and oral availability of the novel ALK/ROS1 inhibitor lorlatinib. European Journal of Pharmaceutics and Biopharmaceutics. 2019, 136: 120-130 [3]Spatari C, Li W, Schinkel A H, et al. Bioanalytical assay for the quantification of the ALK inhibitor lorlatinib in mouse plasma using liquid chromatography-tandem mass spectrometry. Journal of Chromatography B. 2018, 1083: 204-208 [4]Sparidans R W, Li W, Schinkel A H, et al. Bioanalytical liquid chromatography-tandem mass spectrometric assay for the quantification of the ALK inhibitors alectinib, brigatinib and lorlatinib in plasma and mouse tissue homogenates. Journal of Pharmaceutical and Biomedical Analysis. 2018 Nov 30;161:136-143

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