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Purity: ≥98%
PD173074 (PD-173074) is a novel, potent, cell permeable and selective FGFR1 inhibitor with potential antineoplastic activity. In cell-free experiments, it inhibits VEGFR2 with an IC50 of 100-200 nM and FGFR1 with an IC50 of ~25 nM. It also exhibits ~1000-fold selectivity for FGFR1 over PDGFR and c-Src. In vitro, PD173074 demonstrates strong anti-proliferative activity against a variety of cancer cell lines, including UM-UC-14 and MGHU3, which expressed FGFR3 protein mutations.
| Targets |
FGFR1 (IC50 = 25 nM); VEGFR2 (IC50 = 100 nM)
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| ln Vitro |
PD173074 is an FGFR1 ATP-competitive inhibitor with a Ki of approximately 40 nM. Another potent VEGFR2 inhibitor is PD173074. With 1000-fold or higher IC50 values, PD173074 weakly inhibits the activities of Src, InsR, EGFR, PDGFR, MEK, and PKC in comparison to FGFR1. With an IC50 of 1–5 nM and 100–200 nM, respectively, PD173074 inhibits the autophosphorylation of FGFR1 and VEGFR2 in a dose-dependent manner.[1] With an IC50 of 12 nM, PD173074 exhibits dose-dependent inhibition of FGF-2 promotion of granule neuron survival, 1,000 times more potent than SU 5402.[2] In oligodendrocyte (OL) lineage cells, PD173074 specifically blocks the effects of FGF-2 on proliferation, differentiation, and MAPK activation.[3] PD173074 exhibits efficacy against both FGFR3 mutations and the WT receptor in multiple myeloma (MM) cell lines. Moreover, PD173074 has an IC50 of approximately 5 nM and strongly suppresses the dose-dependent autophosphorylation of FGFR3. With an IC50 of less than 20 nM, PD173074 treatment significantly lowers the viability of FGFR3-expressing KMS11 and KMS18 cells. The expression of FGFR3 is closely associated with PD173074's inhibition of aFGF-stimulated MM cell growth. Treatment with PD173074 totally eliminates NIH 3T3 transformation caused by Y373C FGFR3, but not Ras V12. This indicates that PD173074 does not have a non-specific cytotoxic effect and instead targets FGFR3-mediated cell transformation. KMS11 and KMS18 cells are also induced to functionally mature by PD173074.[4]
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| ln Vivo |
PD173074 can be administered to mice at a dose-dependent rate of either 1 mg/kg/day or 2 mg/ka/day to effectively block angiogenesis induced by VEGF or FGF, with no apparent toxicity.[1] PD173074 prevents NIH 3T3 cells transfected with mutant FGFR3 from growing in vivo in nude mice. In a KMS11 xenograft myeloma model, inhibiting FGFR3 with PD173074 slows the growth of tumors and improves mice survival.[4] Compared to control sham-treated animals, oral administration of PD173074 in the H-510 xenograft increases median survival by blocking tumor growth in a manner similar to that observed with single-agent cisplatin administration. Fifty percent of the mice receiving PD173074 in H-69 xenografts experience full responses that last longer than six months. These effects do not result from disruption of the tumor vasculature; rather, they are correlated with increased apoptosis in removed tumors.[5]
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| Enzyme Assay |
In assays with the full-length FGFR-1 kinase, a total volume of 100 μL is used. It contains the following concentrations: 750 μg/mL of a random copolymer of glutamic acid and tyrosine (4:1), different concentrations of PD173074, 60 to 75 ng of enzyme, and 150 mM NaCl, 10 mM MnCl2, 0.2 mM sodium orthovanadate. [γ-32P]ATP (5 μM ATP containing 0.4 μCi of [γ-32P]ATP per incubation) is added to start the reaction, and samples are incubated for 10 minutes at 25°C. Thirty percent trichloroacetic acid is added to stop the reaction, and the material precipitates onto glass-fiber filter mats. The incorporation of [32P] into the glutamate tyrosine polymer substrate is measured by counting the radioactivity retained on the filters in a Wallac 1250 betaplate reader after the filters are cleaned three times with 15% trichloroacetic acid. The radioactivity that remains on the filters after samples without enzyme are incubated is known as nonspecific activity. Total activity (enzyme plus buffer) less nonspecific activity is the formula for calculating specific activity. An IC50 chart is used to calculate the concentration of PD173074 that inhibits FGFR-1 enzymatic activity by 50%.
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| Cell Assay |
There has been prior description of an NIH 3T3 cell line that overexpresses VEGFR2 (Flk-1). Additionally, this cell line naturally expresses FGFR1. In 10 cm2 plates, 1×106 cells grown in DMEM enhanced with 10% calf serum are seeded and given 48 hours to grow. The cells are then put in starvation medium (DMEM with 0.1% calf serum) to become quiescent after the medium is removed. PD 173074 prepared in starvation medium is added to the cells at different concentrations and incubated for 5 minutes after 18 hours. Next, for five minutes at 37°C, the cells are stimulated with a growth factor [100 ng/mL of VEGF or 100 ng/mL of aFGF and 10 µg/mL of heparin]. Following an ice-cold PBS wash, the cells are lysed in 1 mL of lysis buffer that contains phosphatase inhibitor (0.2 mM Na3VO4) and contains 25 mM HEPES pH 7.5, 150 mM NaCl, 1% Triton X-100, 10% glycerol, 1 mM EGTA, 1.5 mM MgCl2, 1 mM PMSF, and 10 µg/mL aprotonin and 10 µg/mL leupeptin. Cell lysates are immunoprecipitated using FGFR1 antibodies for FGFR1 inhibition studies. Phosphotyrosine-specific antibodies are then used for SDS-PAGE and immunoblotting analysis. Cell lysates (20 µL) are subjected to direct SDS-PAGE analysis and immunoblotting using phosphotyrosine-specific antibodies in order to study the inhibition of VEGFR2.
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| Animal Protocol |
Subcutaneous inoculation with 3×105 NIH 3T3 cells expressing Y373C FGFR3 and Ras V12 is performed on six-week-old athymic nude mice. The intraperitoneal injection of 0.05 mol/L lactic acid buffer or 20 mg/kg PD173074 is started on the day of the tumor injection and is administered for nine days. For every experiment, ten mice are used.
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| References |
| Molecular Formula |
C28H41N7O3
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| Molecular Weight |
523.67
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| Exact Mass |
523.33
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| Elemental Analysis |
C, 64.22; H, 7.89; N, 18.72; O, 9.17
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| CAS # |
219580-11-7
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| Related CAS # |
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| Appearance |
Solid powder
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| SMILES |
CCN(CC)CCCCNC1=NC2=NC(=C(C=C2C=N1)C3=CC(=CC(=C3)OC)OC)NC(=O)NC(C)(C)C
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| InChi Key |
DXCUKNQANPLTEJ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C28H41N7O3/c1-8-35(9-2)13-11-10-12-29-26-30-18-20-16-23(19-14-21(37-6)17-22(15-19)38-7)25(31-24(20)32-26)33-27(36)34-28(3,4)5/h14-18H,8-13H2,1-7H3,(H3,29,30,31,32,33,34,36)
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| Chemical Name |
1-tert-butyl-3-[2-[4-(diethylamino)butylamino]-6-(3,5-dimethoxyphenyl)pyrido[2,3-d]pyrimidin-7-yl]urea
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| Synonyms |
PD 173074; PD-173074; PD173074
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.77 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (4.77 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (3.97 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: 5% DMSO+corn oil: 15mg/mL |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9096 mL | 9.5480 mL | 19.0960 mL | |
| 5 mM | 0.3819 mL | 1.9096 mL | 3.8192 mL | |
| 10 mM | 0.1910 mL | 0.9548 mL | 1.9096 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
The expression of MYC and FGFR3 was analyzed by Western blotting in lysates from MGH‐U3 and RT112 cells transfected for 72h withMYCsiRNAs.
TheMYCaccumulation induced by activatedFGFR3 confers sensitivity toBETbromodomain inhibitors inFGFR3‐dependent bladder cancer cellsinvitroandinvivo.EMBO Mol Med.2018 Apr;10(4). pii: e8163. |
MGH‐U3 and RT112 cells were treated with control DMSO, PD [PD173074 (FGFR inhibitor)], SB [SB203580 (p38 inhibitor)] or LY [LY294002 (PI3 kinase inhibitor)] for 72h and cell viability was then assessed by measuring MTT incorporation.
Venn diagram showing the number of upstream regulators (transcription factors) significantly predicted by Ingenuity Pathway Analysis to be involved in the regulation of gene expression observed afterFGFR3knockdown in RT112 and MGH‐U3 cells (left panel).EMBO Mol Med.2018 Apr;10(4). pii: e8163.
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TheMYCaccumulation induced by activatedFGFR3 is dependent on the activation of p38 andAKT.
MYCandFGFR3 are involved in a positive feedback loop in bladder cancer cell lines expressing an activated form ofFGFR3.EMBO Mol Med.2018 Apr;10(4). pii: e8163. |
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