PD173074 showed model-dependent activity in a 72-hour cancer cell viability panel: reported EC50 values were 0.022 μM in KATO III, 0.05 μM in RT4 and 0.12 μM in SNU-16, compared with >5 μM in VM-CUB1, NCI-H1703 and JHH-7. [1]
In CA1 and H1 human embryonic stem cells, a six-day combination of BMP4, A83-01 and 0.1 μM PD173074 induced some trophoblast-associated markers but did not reproduce the full first-trimester trophoblast marker profile assessed by the authors. Low C19MC microRNAs and retained classical HLA expression limit lineage interpretation. These findings apply to the combination, without isolating the contribution of PD173074. [3]
Reversible inhibition of inducible FGFR1 signaling
Cell-based study · Serum-starved MCF10A human mammary epithelial cells expressing inducible FGFR1 (iFGFR1). [1]
At 20 nM, PD173074 strongly suppressed AP20187-induced receptor tyrosine phosphorylation and ERK1/2 phosphorylation. Activity was largely lost following washout and a 6-hour drug-free interval, whereas FIIN-1 retained inhibition. Receptor/ERK abundance and tubulin blots accompany the phospho readouts.
Literature-Reported Treatment Conditions
- Treatment concentration
- 20 nM PD173074; the washout panel also includes 2 nM.
- Exposure duration
- 30-minute inhibitor exposure; washout followed by 6 hours in serum-free medium and 30-minute stimulation.
- Method
- Immunoprecipitation and immunoblotting; matched no-washout signaling comparison.
- Vehicle
- DMSO control; final percentage not stated in the selected methods.
- Co-treatment
- AP20187 (100 nM) activates the engineered receptor; FIIN-1 is a separate comparator.
- Reported limits
- Biological replicate count and quantitative densitometry not stated for these panels.
Methods and source locations
Compound reported in the paper: PD173074; Compound label in the paper: PD173074
Evidence applies to the reported experimental material and conditions; no inference of current Adooq lot performance.
Figure 3C–D and captions; Results: A biotin-labeled FIIN-1; Experimental Procedures: Washout experiment
iFGFR1 tyrosine phosphorylation and downstream ERK1/2 phosphorylation: Anti-phosphotyrosine after anti-HA immunoprecipitation; phospho-ERK1/2 blot; DMSO ± AP20187 controls and total protein/loading controls. — Figure 3C, caption and associated Results
Recovery of iFGFR1 and ERK1/2 phosphorylation after drug removal: Comparison with sustained FIIN-1 inhibition; no inhibitor controls and receptor/ERK/loading controls. — Figure 3D; Experimental Procedures: Washout experiment
Variable cancer cell viability responses
Cell-based study · Human cancer cell panel; representative sensitive gastric/bladder models and insensitive bladder/lung/liver models. [1]
The 72-hour viability panel shows substantial model dependence. KATO III, RT4 and SNU-16 were sensitive, while VM-CUB1, NCI-H1703 and JHH-7 had reported PD173074 EC50 values above 5 μM. These are cellular viability endpoints, not purified kinase IC50 values.
Literature-Reported Treatment Conditions
- Treatment concentration
- Concentration-response testing; full dose series not stated for the Table 2 retest.
- Exposure duration
- 72 hours.
- Method
- Syto-60 staining; values expressed relative to untreated control wells; EC50 fitted with GraphPad Prism 4.
- Vehicle
- Untreated control wells; final vehicle percentage not stated for this assay.
- Co-treatment
- PD173074 and FIIN-1 were tested separately.
- Reported limits
- Replicate counts and uncertainty estimates for Table 2 EC50 values are not stated in the read table/methods.
Methods and source locations
Compound reported in the paper: PD173074; Compound label in the paper: PD173074
Evidence applies to the reported experimental material and conditions; no inference of current Adooq lot performance.
Table 2 and caption; Results: FIIN-1 inhibits proliferation of cancer cells; Experimental Procedures: Cell viability assays
Cellular viability after 72 hours: Syto-60 staining relative to untreated controls. — Table 2: KATO III, RT4 and SNU-16 rows; Cell viability assays
Cellular viability after 72 hours: Same panel and endpoint as sensitive models; operator retained exactly. — Table 2: VM-CUB1, NCI-H1703 and JHH-7 rows; Cell viability assays
FGFR3 kinase-domain interaction by NMR
Biochemical assay · Purified wild-type human FGFR3 kinase domain, residues 455–768, compared in ligand-free and PD173074-bound states. [2]
PD173074 binding altered backbone amide chemical shifts, particularly in the P-loop, hinge and DFG-latch regions. These data support a direct interaction with the FGFR3 kinase domain; they do not provide an enzyme IC50 or an affinity constant.
Literature-Reported Treatment Conditions
- Method
- TROSY-based NMR and chemical-shift perturbation mapping. Protein samples were 77–230 μM; this range is not a PD173074 treatment concentration.
- Vehicle
- PD173074 stock prepared in DMSO.
- Co-treatment
- None; ligand-free protein served as comparator.
- Reported limits
- A single final PD173074 concentration and incubation duration are not provided for the adopted qualitative comparison. Free and bound assignment spectra were acquired at 298 K and 303 K, respectively.
Methods and source locations
Compound reported in the paper: PD173074; Compound label in the paper: PD173074
Evidence applies to the reported experimental material and conditions; no inference of current Adooq lot performance.
Figures 1–2 and captions; Methods and experiments: NMR spectroscopy and data processing; Assignment and data deposition
Backbone amide chemical-shift perturbations on inhibitor interaction: Comparison of apo and inhibitor-complex spectra; mapping onto a homology model, not a newly solved inhibitor co-crystal structure. — Figure 2 and caption; Assignment and data deposition
Limits of trophoblast identity after BAP differentiation
Cell-based study · CA1 and H1 human embryonic stem cells, with FGF2-maintained controls and JEG-3 marker comparator. [3]
A six-day BMP4/A83-01/PD173074 combination induced KRT7 and reduced pluripotency markers, but the resulting cells did not reproduce the full set of first-trimester trophoblast markers used by the authors. C19MC miRNAs remained low and HLA-A/HLA-B expression was retained. This is a combination study and cannot establish a PD173074-alone differentiation effect.
Literature-Reported Treatment Conditions
- Treatment concentration
- 0.1 μM PD173074.
- Exposure duration
- 6 days; harvested on day 6.
- Method
- Differentiation in mouse embryonic fibroblast-conditioned medium; immunostaining, qRT-PCR, methylation analysis and flow cytometry.
- Vehicle
- FGF2 control received a matched amount of DMSO.
- Co-treatment
- BAP: BMP4 10 ng/mL plus A83-01 1 μM plus PD173074 0.1 μM; control: FGF2 4 ng/mL.
- Reported limits
- No PD173074-alone arm; salt/solvate and lot identity not specified.
Methods and source locations
Compound reported in the paper: PD173074; Compound label in the paper: PD173074
Evidence applies to the reported experimental material and conditions; no inference of current Adooq lot performance.
Results: BMP-Treated hESC; Figures 4–5 and captions; Document S1, culture conditions table (PDF p.7) and differentiation protocol (p.8)
Protein and transcript marker profile: Immunofluorescence and qRT-PCR; n = 3 independent experiments for Figure 4A–B; Figure 4B error bars are SE. — Figure 4A–B and caption; Results: BMP-Treated hESC
C19MC microRNA expression: qRT-PCR normalized to miR-103a; n = 3 independent experiments; error bars are SE; mammary gland negative control. — Figure 5A and caption; Results: BMP-Treated hESC
HLA class I surface profile: Flow cytometry with isotype controls; n = 3 independent experiments. — Figure 5B–E and caption; Results: BMP-Treated hESC
