In an EGFR-inhibitor comparison, 200 nM pelitinib for 12 hours reduced FLIP-L protein in PC-9 cells relative to DMSO. HCC827 cells did not show a comparable marked reduction in the displayed blot. Pelitinib-specific apoptosis or proteasomal-degradation conclusions were not established by this comparison. [2]
In SK-Hep1 cells, 1 µM EKB-569 for 24 hours increased the reported G0/G1 fraction from 72.8% to 86.4%. SNU-475 showed a smaller change, from 75.8% to 81.2%, at 5 µM. [1]
In a 48-hour MTS assay, pelitinib growth-inhibition IC50 values were 3.14 µM in HAP1 wild-type cells, 2.03 µM in PWWP2B-knockout cells and 0.71 µM in RNF43-knockout cells. These leukemia-derived cell models provide cellular response data rather than EGFR enzyme potency. [3]
Pelitinib comparator and c-FLIP protein in EGFR-mutant NSCLC
Cell-based study · PC-9 and HCC827 NSCLC cells with EGFR exon 19 deletion [2]
In Fig. 1D, the pelitinib lane shows a marked decrease of FLIP-L in PC-9 relative to DMSO, whereas HCC827 does not show a comparable marked decrease. This is a qualitative comparator observation; the main mechanistic and apoptosis experiments used osimertinib.
Literature-Reported Treatment Conditions
- Treatment concentration
- 200 nM
- Exposure duration
- 12 h
- Method
- Western blot; DMSO control and actin loading control. Other EGFR inhibitors were separate comparator treatments.
- Vehicle
- DMSO
- Reported limits
- Pelitinib salt/form, catalog number, lot and blot replication count are not specified for panel D; no densitometry or pelitinib IC50 is supplied there.
Methods and source locations
Compound reported in the paper: Pelitinib (EKB-569); Compound label in the paper: Pelitinib
Evidence applies to the named compound and tested preparation/model. It does not establish equivalence of an Adooq lot or an unspecified salt.
Fig. 1D and caption; Materials and Methods: Reagents; Results first subsection
FLIP-L protein relative to DMSO in PC-9 and HCC827: Western blot with actin loading control — Fig. 1D, pelitinib and DMSO lanes; caption: 200 nM, 12 h
Cell-cycle response to EKB-569
Cell-based study · SK-Hep1 cells; SNU-475 limiting comparator reported in Results [1]
EKB-569 increased the G0/G1 fraction in SK-Hep1. The paper reports a smaller shift in SNU-475 at a higher concentration.
Literature-Reported Treatment Conditions
- Treatment concentration
- SK-Hep1: 1 µM; SNU-475: 5 µM (reported comparison)
- Exposure duration
- 24 h
- Method
- Propidium iodide DNA staining and flow cytometry; untreated SK-Hep1 comparator is shown. SNU-475 numerical comparison is reported in Results but not plotted in Fig. 1.
- Reported limits
- Vehicle and independent replicate count/error estimates for the cell-cycle experiment are not specified.
Methods and source locations
Compound reported in the paper: Pelitinib (EKB-569); Compound label in the paper: EKB-569
Evidence applies to the named compound and tested preparation/model. It does not establish equivalence of an Adooq lot or an unspecified salt.
Fig. 1 image/caption; Results: Increase of G0-G1 arrest by EKB-569; Methods: Cell cycle analysis; Chemicals and antibodies
G0/G1 fraction in SK-Hep1: Flow cytometry; values reported in Results and visually checked against Fig. 1 — Results: Increase of G0-G1 arrest; Fig. 1
G0/G1 fraction in SNU-475: Flow cytometry; text-reported limiting comparison, not shown in the Fig. 1 image — Results: Increase of G0-G1 arrest by EKB-569
Model-dependent ERK and AKT phosphorylation response
Cell-based study · SK-Hep1 and SNU-475 cells [1]
After 24 h, phosphorylated ERK and AKT bands decreased with EKB-569 concentration in SK-Hep1. SNU-475 did not display the same concentration-dependent suppression. Total ERK/AKT and β-actin controls are displayed.
Literature-Reported Treatment Conditions
- Treatment concentration
- 0, 1, 2.5 and 5 µM
- Exposure duration
- 24 h
- Method
- Western blot of phosphorylated and total ERK/AKT, CDK4, p21 and β-actin; 0 µM comparator.
- Reported limits
- Vehicle is not specified. No numerical densitometry/IC50 is given for these blots.
Methods and source locations
Compound reported in the paper: Pelitinib (EKB-569); Compound label in the paper: EKB-569
Evidence applies to the named compound and tested preparation/model. It does not establish equivalence of an Adooq lot or an unspecified salt.
Fig. 2 image/caption; Results: EKB-569 decreased the phosphorylation of AKT and ERK; Methods: Western blot analysis and Chemicals and antibodies
Phosphorylated ERK and AKT versus total proteins and β-actin: Western blot; at least two independently prepared lysates per protein, per caption — Fig. 2; Results corresponding subsection
Pelitinib growth-inhibition response across HAP1 genotypes
Cell-based study · HAP1 wild type, HAP1 PWWP2B knockout and HAP1 RNF43 knockout; leukemia-derived near-haploid cells [3]
Table 2 reports cellular growth-inhibition IC50 values of 3.14, 2.03 and 0.71 µM in wild-type, PWWP2B-knockout and RNF43-knockout cells, respectively. These are not EGFR enzyme IC50 values or measurements in gastric cancer cells.
Literature-Reported Treatment Conditions
- Treatment concentration
- 0.3125, 0.625, 1.25, 2.5, 5, 10, 20 and 40 µM
- Exposure duration
- 48 h
- Method
- MTS cell growth/viability assay; untreated controls; nonlinear dose-response fitting using GraphPad Prism.
- Reported limits
- Solvent and exact pelitinib salt/lot are not specified. Table 2 gives point IC50 estimates without uncertainty.
Methods and source locations
Compound reported in the paper: Pelitinib (EKB-569); Compound label in the paper: Pelitinib
Evidence applies to the named compound and tested preparation/model. It does not establish equivalence of an Adooq lot or an unspecified salt.
Table 2, Pelitinib row and genotype headers; Methods: Confirmatory growth inhibition assays and High-throughput drug screening; Results: FDA-approved drug library screen
Growth-inhibition IC50 in HAP1 wild type / PWWP2B KO / RNF43 KO: 48-h MTS assay; Results reports n = 3 for dose-response experiments; Table 2 contains no IC50 uncertainty — Table 2, Pelitinib row; Methods: Confirmatory growth inhibition assays
