small-molecule fgfr inhibitors  (MultiTarget Pharmaceuticals)

Journal: Molecular Cancer

Article Title: Clinical advances and challenges in targeting FGF/FGFR signaling in lung cancer

doi: 10.1186/s12943-024-02167-9

Figure Lengend Snippet: FGFR genomic alterations in lung cancer

Article Snippet: These approaches include small-molecule FGFR inhibitors that target the ATP-binding site of the tyrosine kinase domain of various growth factor receptors (multitarget TKIs), selective TKIs that specifically target the kinase domain of FGFRs, and biological agents (Fig. ).

Techniques: Mutagenesis

Journal: Molecular Cancer

Article Title: Clinical advances and challenges in targeting FGF/FGFR signaling in lung cancer

doi: 10.1186/s12943-024-02167-9

Figure Lengend Snippet: Targeting FGF/FGFR signaling in lung cancer. Drugs targeting FGFR that are at the clinical stage include multitarget TKIs (nintedanib, dovitinib, pazopanib, and ponatinib) and selective TKIs (pan-FGFR inhibitors such as erdafitinib and rogaratinib; FGFR1/2/3 inhibitors such as pemigatinib, infigratinib and fexagratinib). FP-1039, which targets FGF, has also been investigated in clinical trials. New strategies for targeting FGF/FGFR include the use of FGFR degraders (DGY-09-192 and LC-MB12), specific FGFR inhibitors and anti-FGF2 aptamers

Article Snippet: These approaches include small-molecule FGFR inhibitors that target the ATP-binding site of the tyrosine kinase domain of various growth factor receptors (multitarget TKIs), selective TKIs that specifically target the kinase domain of FGFRs, and biological agents (Fig. ).

Techniques: Clinical Proteomics

Journal: Molecular Cancer

Article Title: Clinical advances and challenges in targeting FGF/FGFR signaling in lung cancer

doi: 10.1186/s12943-024-02167-9

Figure Lengend Snippet: Clinical trials of multitarget TKIs in lung cancer

Article Snippet: These approaches include small-molecule FGFR inhibitors that target the ATP-binding site of the tyrosine kinase domain of various growth factor receptors (multitarget TKIs), selective TKIs that specifically target the kinase domain of FGFRs, and biological agents (Fig. ).

Techniques: Clinical Proteomics, Control, Amplification, Adjuvant

Journal: Molecular Cancer

Article Title: Clinical advances and challenges in targeting FGF/FGFR signaling in lung cancer

doi: 10.1186/s12943-024-02167-9

Figure Lengend Snippet: Clinical trials of selective FGFR inhibitors in lung cancer

Article Snippet: These approaches include small-molecule FGFR inhibitors that target the ATP-binding site of the tyrosine kinase domain of various growth factor receptors (multitarget TKIs), selective TKIs that specifically target the kinase domain of FGFRs, and biological agents (Fig. ).

Techniques: Clinical Proteomics

Journal: Molecular Cancer

Article Title: Clinical advances and challenges in targeting FGF/FGFR signaling in lung cancer

doi: 10.1186/s12943-024-02167-9

Figure Lengend Snippet: Resistance mechanisms of targeted FGFR therapies in lung cancer. In addition to FGFR mutations, changes in signaling pathways and the tumor microenvironment confer resistance to targeted FGFR therapies in lung cancer. ( A ) Bypass and downstream pathway activation. ( B ) Multidrug resistance pathway activation and DNA damage inhibition. ( C ) Cytokine reprogramming in the tumor microenvironment

Article Snippet: These approaches include small-molecule FGFR inhibitors that target the ATP-binding site of the tyrosine kinase domain of various growth factor receptors (multitarget TKIs), selective TKIs that specifically target the kinase domain of FGFRs, and biological agents (Fig. ).

Techniques: Protein-Protein interactions, Activation Assay, Inhibition

Journal: Cancer Research

Article Title: CRISPR-Mediated Kinome Editing Prioritizes a Synergistic Combination Therapy for FGFR1-Amplified Lung Cancer

doi: 10.1158/0008-5472.can-20-2276

Figure Lengend Snippet: Figure 1. CRISPR/Cas9 knockout screens to iden- tify synthetic lethal targets in FGFR1- amplified lung cancer cells treated with the FGFR inhibitor AZD4547. A, Sche- matic of the timeline and experimental procedures of CRISPR/Cas9 screen using a pooled sgRNA library targeting the human kinome. B, Box plots of log2- transformed sgRNA normalized read counts from H520 cells before drug treatment (baseline) after 21-day treat- ment with vehicle (vehicle) or AZD4547 (AZD4547), with experimental repli- cates (n ¼ 3) indicated (Baseline_1/2/ 3, Vehicle_1/2/3, and AZD4547_1/2/3). C, Scatterplots showing log2-trans- formed sgRNA normalized read counts of AZD4547- versus vehicle-treated H520 cells, with sgRNAs targeting PIP4K2C, PLK1, and CSNK2A1 highlight- ed. Although PIP4K2C is a substrate of mTOR previously shown to be a syn- thetic lethal partner with FGFR1, PLK1 and CSNK2A1 (encoding CK2a) have been reported to function in the same process (DNA damage repair), high- lighting the potential of PLK1 and CSNK2A1 as novel synthetic lethal genes with FGFR1. D, Frequency histograms showing the top screen hits (n ¼ 7; ranked by log2FC) negatively selected in vehicle versus baseline (left) and AZD4547 versus vehicle (right) after day 21. Individual sgRNAs targeting the indicated genes are highlighted by red lines, with the log2FC and P values indicated.

Article Snippet: Small-molecule inhibitors targeting FGFR (AZD4547, BGJ398), PLK1 (BI2536, BI6267), microtubule (paclitaxel), and other RTKs were purchased from Selleckchem and shown in Supplementary Table S2.

Techniques: CRISPR, Knock-Out, Transformation Assay

Journal: Cancer Research

Article Title: CRISPR-Mediated Kinome Editing Prioritizes a Synergistic Combination Therapy for FGFR1-Amplified Lung Cancer

doi: 10.1158/0008-5472.can-20-2276

Figure Lengend Snippet: Figure 2. Genetic and pharmacological antago- nism of PLK1 plus FGFR inhibitors syn- ergistically suppresses proliferation and enhances apoptosis in FGFR1-amplified lung cancer cells. A, Immunoblots of H1703 and H520 cells treated for 2hours with AZD4547 as indicated concentra- tion.B,Immunoblots ofH1703and H520 cells transfected with PLK1-specific siRNAs (si-PLK1) or scramble control siRNAs (si-Control). C, Dose–response curves of H1703 and H520 cells ex- pressing PLK1 or control siRNAs to AZD4547.Cellproliferationwasassayed 72 hours after drug treatment (84 hours after siRNA transfection). Data are pre- sented as mean of three independent experiments (n ¼ 3). D, H1703 and H520 cells expressing PLK1 or control siRNAs weretreatedwithAZD4547for72hours and cultured in drug-free medium for additional 7–14 days. Surviving cells afterthe treatmentwerefixedand visu- alized by crystal violet staining. Repre- sentative images of three independent experiments (n ¼ 3) are shown. E, H1703 and H520 cells expressing PLK1 or control siRNAs were treated with AZD4547 for 72 hours before anal- ysis by apoptotic assay. Data are pre- sented as mean SD (n ¼ 3). , P < 0.01 and , P < 0.001 by two-way ANOVA with Tukey multiple compari- sons test. ns, not significant. F–I, The PLK1 inhibitor BI2536 synergistically enhances the antiproliferative effect of AZD4547 in FGFR1-amplified lung can- cer cells. H1703 and H520 cells treated for 72 hours with AZD4547 and BI2536, aloneorincombination,weresubjected to proliferation analysis (F and H) and clonogenic assay (G and I). The plot of fraction affected (Fa) and combination index (CI) are shown underneath, with CI< 1.0 indicatingsynergistic effect.The heatmap (G and I; right) indicates the percentage of viable cells after the treatment, based on quantification of clonogenic results (right). Data are presented as mean SD (n ¼ 3). , P < 0.001; , P < 0.0001 by two-way ANOVA and Tukey post hoc test. J and K, H1703 (J) and H520 (K) cells treated with the indicated drugs for 24, 48, and 72 hours were subjected to flow cytometry–based apoptosis assay. Data are presented as mean of threeindependent experiments (n ¼ 3).

Article Snippet: Small-molecule inhibitors targeting FGFR (AZD4547, BGJ398), PLK1 (BI2536, BI6267), microtubule (paclitaxel), and other RTKs were purchased from Selleckchem and shown in Supplementary Table S2.

Techniques: Western Blot, Transfection, Control, Expressing, Cell Culture, Staining, Clonogenic Assay, Cytometry, Apoptosis Assay

Journal: Cancer Research

Article Title: CRISPR-Mediated Kinome Editing Prioritizes a Synergistic Combination Therapy for FGFR1-Amplified Lung Cancer

doi: 10.1158/0008-5472.can-20-2276

Figure Lengend Snippet: Figure 3. Combined FGFR and PLK1 inhibition enhances G2–M arrest and DNA damage- induced apoptosis by activating the CHK2/ E2F1 axis. A, Gene set enrichment analysis of H520 and H1581 cells treated with CH5183284/Debio 1347, a selective FGFR inhibitor. Transcriptomic gene expression data are based on the GEO dataset GSE73024. B, Cell-cycle analysis of H1703 cells treated with AZD4547 (5 mmol/L) and BI2536 (5 nmol/L), alone or in combination, for 24, 48, and 72 hours. Data are presented as mean of three inde- pendent experiments (n ¼ 3). C, Immuno- blots of H1703 cells treated with vehicle (DMSO), AZD4547, and BI2536, alone or in combination, for 6 and 24 hours. D, Immunofluorescence of H1703 cells treated with vehicle (DMSO), AZD4547 (5 mmol/L), and BI2536 (5 nmol/L), alone or in combination, for 24 hours. Cells were subsequently stained with antibodies against gH2AX (red) and pHH3 (green) and DAPI (blue; nuclei). Scale bar (white), 25 mm. E, Immunoblots of H1703 cells transfected with E2F1 or control siRNAs and subsequently treated with vehicle () or AZD4547 (5 mmol/L) plus BI2536 (5 nmol/L) for 24 hours. F, H1703 cells transfected with E2F1 or control siRNAs were treated with vehicle (DMSO), AZD4547 (5 mmol/L), BI2536 (5 nmol/L), and the drug combination for 48 hours and subsequently analyzed by apoptotic assay. Data are presented as mean SD (n ¼ 3). , P < 0.001 by two-way ANOVA with Tukey multiple comparisons test.

Article Snippet: Small-molecule inhibitors targeting FGFR (AZD4547, BGJ398), PLK1 (BI2536, BI6267), microtubule (paclitaxel), and other RTKs were purchased from Selleckchem and shown in Supplementary Table S2.

Techniques: Inhibition, Gene Expression, Cell Cycle Assay, Western Blot, Staining, Transfection, Control

Journal: Cancer Research

Article Title: CRISPR-Mediated Kinome Editing Prioritizes a Synergistic Combination Therapy for FGFR1-Amplified Lung Cancer

doi: 10.1158/0008-5472.can-20-2276

Figure Lengend Snippet: Figure 4. PLK1 promotesacquired resistance to FGFR-targeted therapy. A,Immunoblots of AZD4547-resistant (H1703R, H520R)and parental (H1703, H520)cells after 24 hour treatment with vehicle or the PLK1 inhibitor BI2536 (5 nmol/L). B and C, Dose–response curves (top) and clonogenic assay (middle and bottom) of AZD4547- resistant (H1703R, H520R) and parental (H1703, H520) cells to BI2536. Cellswere treated for 72 hours before subjected to proliferation and clonogenicassay.Data are presented as mean SD (n ¼ 3), with representative results and quantification of clonogenic assay shown. , P < 0.05; , P < 0.01; , P < 0.001; , P < 0.0001 by one-way ANOVA with Tukey multiple comparisons test. D, AZD4547-resistant (H1703R, H520R) and parental (H1703, H520) cells treated for 72 hours with BI2536 (5 nmol/L) were analyzed by flow cytometry–based apoptosis assay. The percentage of apoptotic cells was determined by the ratio of apoptotic populations (Annexin Vþ/PI plus Annexin Vþ/PIþ) versus the total cells. Data are presented as mean SD (n ¼ 3). , P < 0.01; , P < 0.001 by one-way ANOVA with Tukey multiple comparisons test. E, Immunoblots of H1703R and H520R cells transfected with E2F1 or control siRNAs. F, H1703R and H520R transfected with E2F1 or control siRNAs were treated with BI2536 (5 nmol/L) for 72 hours and subsequently analyzed by apoptotic assay. Data are presented as mean SD (n ¼ 3). , P < 0.05; , P < 0.01 by one-way ANOVA with Tukey multiple comparisons test. G, Schematic diagram depicting the findings of this study. PLK1 activation promotes DNA damage repair and restrains CHK2 and E2F1 phosphorylation, which compensates for FGFR inhibtion-induced cell-cycle arrest (left). However, combined FGFR1/PLK1 inhibition induces DNA damage, leading to the induction of CHK2 and E2F1 phosphorylation and in turn apoptosis.

Article Snippet: Small-molecule inhibitors targeting FGFR (AZD4547, BGJ398), PLK1 (BI2536, BI6267), microtubule (paclitaxel), and other RTKs were purchased from Selleckchem and shown in Supplementary Table S2.

Techniques: Western Blot, Clonogenic Assay, Cytometry, Apoptosis Assay, Transfection, Control, Activation Assay, Phospho-proteomics, Inhibition

Journal: Cancer Research

Article Title: CRISPR-Mediated Kinome Editing Prioritizes a Synergistic Combination Therapy for FGFR1-Amplified Lung Cancer

doi: 10.1158/0008-5472.can-20-2276

Figure Lengend Snippet: Figure 5. Combined FGFR- and PLK1-targeted therapy potently inhibits FGFR1-amplified lung cancer xenografts. A and B, Growth curve of H1703 (A) and H520 (B) xenografts treated with vehicle, AZD4547 (10 mg/kg/d), BI2536 (40 mg/kg/wk) or BI6727 (5 mg/kg/d), alone and in combination. Data are shown as mean SD. , P < 0.001; , P < 0.0001 by one-way ANOVA with Tukey multiple comparisons test. C–F, Relative tumor volume (C and D) and weights (E and F) of H1703 and H520 xenografts after the treatment for 22 or 24 days. , P < 0.05; , P < 0.01 by one-way ANOVA with Tukey multiple comparisons test. G, Growth curve of an FGFR1-amplified SQLC PDX model (BE937T) treated with vehicle, AZD4547 (10 mg/kg/d), BI6727 (5 mg/kg/d), and the drug combination for the indicated time. Data are shown as mean SD. , P < 0.01 by one-way ANOVA with Tukey multiple comparisons test. H and I, Relative tumor volume (H) and weights (I) of PDX (BE937T) xenografts after the treatment for 21 days. ns, not significant; , P < 0.05 by one-way ANOVA with Tukey multiple comparisons test. J, Hematoxylin and eosin (H&E) and IHC analysis (gH2AX, caspase-3, and Ki-67) of PDX (BE973T) xenografts after the treatment.Original overall magnification, 400. Scalebar,200 mm. K, Quantification of the IHC data (J) for the positivity of gH2AX, caspase-3, and Ki-67. , P < 0.05; , P < 0.01; , P < 0.001; , P < 0.0001 by one-way ANOVA with Tukey multiple comparisons test.

Article Snippet: Small-molecule inhibitors targeting FGFR (AZD4547, BGJ398), PLK1 (BI2536, BI6267), microtubule (paclitaxel), and other RTKs were purchased from Selleckchem and shown in Supplementary Table S2.

Techniques:

Journal: Molecular and Cellular Neurosciences

Article Title: Regional effects of endocannabinoid, BDNF and FGF receptor signalling on neuroblast motility and guidance along the rostral migratory stream

doi: 10.1016/j.mcn.2014.12.001

Figure Lengend Snippet: FGFR signalling regulates neuroblast migration in the RMS. Time-lapse movies of GFP-labelled neuroblasts in the descending arm of the RMS from slices treated with different drugs targeting the FGFR signalling system (1 μM AZD4547 or 1 μg/ml FGFR1-Fc) were analysed using Volocity. Representative migratory tracks of 15 cells over a 3 hour filming period from a control (A), an AZD4547 (B) or a FGFR1-Fc (C) -treated brain slice. White stars mark the end point of each migration track, OB indicates the location of the olfactory bulb. Inhibiting FGF signalling increased the percentage of time spent immobile by neuroblasts (D), while significantly decreasing the percentage of cells migrating towards the OB (E) and the overall cell displacement (F). Graphs show mean ± s.e.m. (n = 6–7 brain slices for each condition, ~ 15–30 cells analysed per slice); *p < 0.05, **p < 0.01.

Article Snippet: Two selective small molecule FGFR inhibitors were used and these were PD173074 (Calbiochem) and AZD4547 (Active Biochem).

Techniques: Migration, Control, Slice Preparation

Journal: Molecular and Cellular Neurosciences

Article Title: Regional effects of endocannabinoid, BDNF and FGF receptor signalling on neuroblast motility and guidance along the rostral migratory stream

doi: 10.1016/j.mcn.2014.12.001

Figure Lengend Snippet: FGF and eCB signalling can operate independently in culture. RMS explants isolated from P5–P8 mouse brains were embedded in Matrigel and left to migrate for either 6 h or 24 h before fixation. Representative pictures of explants stained with phalloidin treated with vehicle (A), 1 μM of the FGFR inhibitor PD173074, (B) or 2 ng/ml FGF-2 (C) taken 24 h after embedding. In the 6 h assay, treatment with 1 μM PD173074 significantly decreased migration to the same extent observed after incubation with the CB1/2 antagonists AM251 and JTE-907 (0.5 μM each) (D). When FGFR and CB receptors were inhibited simultaneously, there was no further significant decrease in migration (D). In the 24 h assay FGF-2 (2 ng/ml) significantly promoted migration to the same level observed after incubation with the CB1 agonist ACEA (0.5 μM), and treatment with both did not cause any additive effect on migration (E). In the 24 h assay the CB1 agonist ACEA, the CB2 agonist JWH-133 and the MAGL inhibitor JZL-184 (all at 0.5 μM) could all still significantly stimulate migration in the presence of the FGFR inhibitor PD173074 (1 μM) (F). In the presence of the CB receptor antagonists AM251 and JTE-907 (0.5 μM each), FGF-2 (2 ng/ml) still induced a significant migratory response in the 24 h assay. Graphs show mean ± SEM (n = 4); *p < 0.05, **p < 0.01, ***p < 0.001. Bar, 100 μm for (A–C).

Article Snippet: Two selective small molecule FGFR inhibitors were used and these were PD173074 (Calbiochem) and AZD4547 (Active Biochem).

Techniques: Isolation, Staining, Migration, Incubation

Journal: Molecular and Cellular Neurosciences

Article Title: Regional effects of endocannabinoid, BDNF and FGF receptor signalling on neuroblast motility and guidance along the rostral migratory stream

doi: 10.1016/j.mcn.2014.12.001

Figure Lengend Snippet: Signalling through FGFR regulates the morphology of migrating neuroblasts in vivo . P2 mouse pups were electroporated with pCX-EGFP and 5 days later treated with the FGFR inhibitor AZD4547 (12.5 mg/kg I.P., two doses with 12 hour interval). After 24 h, brains were fixed, sliced and stained for GFP. Representative pictures of migrating neuroblasts in animals treated with vehicle (A) and AZD4547 (B) in 4 different regions along the RMS (labelled 1–4 as depicted in the cartoon). Inhibiting FGFR signalling significantly decreased the process length of migrating neuroblasts only in regions 1 and 2 of the RMS (C). Graphs show mean ± s.e.m. (n = 3–4 animals for each condition, 6 consecutive slices were analysed per brain, ~ 100–200 cells analysed per region); *p < 0.05, **p < 0.01. Bar, 50 μm for (A–B).

Article Snippet: Two selective small molecule FGFR inhibitors were used and these were PD173074 (Calbiochem) and AZD4547 (Active Biochem).

Techniques: In Vivo, Staining

Journal: Frontiers in Oncology

Article Title: Targeting Genomic Alterations in Squamous Cell Lung Cancer

doi: 10.3389/fonc.2013.00195

Figure Lengend Snippet: Targetable genes and ongoing clinical trials in squamous cell carcinoma of the lung .

Article Snippet: Genomic and cell line sensitivity studies on cancer cell lines also demonstrated sensitivity of FGFR gene alterations for the pan-FGFR small molecule inhibitor NVP-BGJ398 (Novartis, Basel, Switzerland) ( ).

Techniques: Clinical Proteomics, Amplification, Mutagenesis, Expressing