PF-573228 is an ATP-competitive focal adhesion kinase (FAK) inhibitor (IC50=4nM). PF-573228 binds to the FAK kinase domain and inhibits FAK autophosphorylation at Tyr397. PF-573228 disrupts the formation of the FAK-Src complex and downstream signaling for haptotaxis, cell migration, and focal adhesion turnover. PF-573228 can be used in studies related to tumors (such as small cell lung cancer, endocrine-resistant breast cancer, glioblastoma), cardiac fibrosis, skin fibrosis/hypertrophic scars[1-4].
In vitro, co-treatment of HepG2, Huh7, and MHCC97H cells with PF-573228 and 2.5µM Sorafenib for 1-48 hours inhibited Sorafenib-induced upregulation of FAK phosphorylation at Tyr397. PF-573228 synergized with Sorafenib to reduce HCC cell viability and suppress colony formation[5]. Treatment of H2596 cells, MiaPaca 2 cells, and Capan 1 cells with 1-10µM PF-573228 for 24-72 hours dose-dependently reduced cell viability, induced cell cycle arrest, and increased the proportion of apoptotic cells[6]. Treatment of ARK-1 cells with 10µM PF-573228 for 1 hour reduced FAK phosphorylation levels at Tyr397. Treatment of ARK-1 cell-formed 3D spheroids with 10µM PF-573228 for 48 hours significantly reduced spheroid perimeter, decreased Cyclin D1 levels, and reduced cell migration speed and cumulative migration distance[7].
In vivo, NKG mice bearing subcutaneous xenografts of MGC803-CPZ-OENC or MGC803-CPZ-OE cells reaching approximately 100mm³ received daily intraperitoneal injections of 5mg/kg PF-573228 for 5 days per week for 35 consecutive days. PF-573228 inhibited xenograft tumor growth, with a more pronounced inhibitory effect in the CPZ-OE group compared to the CPZ-OENC group[8]. C57B/6 mice received a single intratracheal instillation of 3U/kg Bleomycin Sulfate to establish a pulmonary fibrosis model, followed by daily intraperitoneal injections of 50mg/kg PF-573228 for 21 consecutive days. PF-573228 reduced lung tissue fibrosis, decreased whole-lung Fibronectin and procollagen-1 protein levels, and reduced α-SMA expression[9]. On day 3 after optic nerve transection, Sprague-Dawley rats received a single intravitreal injection of 25mM PF-573228 (4μL). PF-573228 reduced the density of Fluoro-Gold retrograde-labeled retinal ganglion cells in the retinal ganglion cell layer and decreased the number of surviving injured retinal ganglion cells[10].
References:
[1] Slack-Davis JK, Martin KH, Tilghman RW, et al. Cellular characterization of a novel focal adhesion kinase inhibitor. J Biol Chem. 2007 May 18;282(20):14845-52.
[2] Maher S, Bayachou M, Fu P, et al. Focal adhesion kinase activation is involved in contractile stimulation-induced detrusor muscle contraction in mice. Eur J Pharmacol. 2023 Aug 5;952:175807.
[3] Tsang VSK, Riccio F, Wilson AS, et al. A human iPSC-based neural spheroid platform for modelling glioblastoma infiltration using high-content imaging. Sci Rep. 2025 Dec 13;16(1):1223.
[4] Zhang H, Schaefer A, Wang Y, et al. Gain-of-Function RHOA Mutations Promote Focal Adhesion Kinase Activation and Dependency in Diffuse Gastric Cancer. Cancer Discov. 2020 Feb;10(2):288-305.
[5] Feng J, Ye L, Chen C, et al. RhoE downregulation leads to enhanced cholesterol biosynthesis and sorafenib resistance in hepatocellular carcinoma. J Biol Chem. 2025 Dec;301(12):110918.
[6] Kanteti R, Mirzapoiazova T, Riehm JJ, et al. Focal adhesion kinase a potential therapeutic target for pancreatic cancer and malignant pleural mesothelioma. Cancer Biol Ther. 2018;19(4):316-327.
[7] Lopez-Mejia IC, Pijuan J, Navaridas R, et al. Oxidative stress-induced FAK activation contributes to uterine serous carcinoma aggressiveness. Mol Oncol. 2023;17(1):98-118.
[8] Luo R, Huang W, Wang H, et al. CPZ promotes the proliferation of gastric cancer by activating the Wnt pathway through binding with DVL2. Discover Oncol. 2025;16:2085.
[9] Zhao XK, Cheng Y, Cheng ML, et al. Focal Adhesion Kinase Regulates Fibroblast Migration via Integrin beta-1 and Plays a Central Role in Fibrosis. Sci Rep. 2016 Jan 14;6:19276.
[10] D'Onofrio PM. Integrin Activation and Focal Adhesion Kinase in the Survival and Regeneration of Injured Adult Rat CNS Neurons. MSc Thesis, University of Toronto. 2014.