axl inhibitors sgi-7079  (MultiTarget Pharmaceuticals)

Journal: Cancers

Article Title: Synergistic Inhibition of Drug Resistant KRAS Mutant Non-Small Cell Lung Cancer by Co-Targeting AXL and SRC

doi: 10.3390/cancers17030490

Figure Lengend Snippet: Downstream effects of AXL and SRC in KRAS mutant NSCLC. ( a ) Western blot image showing protein expression of AXL- and SRC-related markers in AXL knockout A549 cells. ( b ) Western blot image showing protein expression for AXL and phospho-SRC in A549 cells treated with either siAXL, Dasatinib or both. ( c ) Western blot showing protein expression of KRAS and phospho-SHP2 (T542 and T580) in A549 cells before or after siAXL or knockout treatments. Increased expression in SHP2 activity is observed when AXL is inhibited. ( d ) Western blot image for downstream KRAS activity in A549 cells treated with either siAXL, Dasatinib or both.

Article Snippet: Dasatinib and AXL inhibitor, SGI-7079, were purchased from Selleck chemicals (Houston, TX, USA).

Techniques: Mutagenesis, Western Blot, Expressing, Knock-Out, Activity Assay

Journal: Cancers

Article Title: Synergistic Inhibition of Drug Resistant KRAS Mutant Non-Small Cell Lung Cancer by Co-Targeting AXL and SRC

doi: 10.3390/cancers17030490

Figure Lengend Snippet: Effect of co-inhibiting AXL and SRC in KRAS mutant NSCLC. ( a ) Western blot image for apoptotic proteins in A549 cells treated with either siAXL, Dasatinib or both. ( b ) Cytochrome-C assay for A549 cells; scale bar = 100 µm. ( c ) Cell-Death assay: Bright field Microscopy images (20×) for triggered cell death in A549 cells before or after either siAXL or DAS or treatment with both; scale bar = 200 µm. ( d ) Cell-Death assay: Bright field microscopy images (20×) of H460AK cells (i) before and (ii) after treatment with Dasatinib (DAS) for 48 h; scale bar = 200 µm. Results indicate higher cell death after DAS treatment in AXL knockout H460 cells.

Article Snippet: Dasatinib and AXL inhibitor, SGI-7079, were purchased from Selleck chemicals (Houston, TX, USA).

Techniques: Mutagenesis, Western Blot, Microscopy, Knock-Out

Journal: Cancers

Article Title: Synergistic Inhibition of Drug Resistant KRAS Mutant Non-Small Cell Lung Cancer by Co-Targeting AXL and SRC

doi: 10.3390/cancers17030490

Figure Lengend Snippet: Synergistic AXL and SRC therapy in vitro. ( a ) Dasatinib (IC 50 ) sensitization of NSCLC cell lines after co-inhibition of AXL + SRC using MTT assay (72 h). ( b ) Dasatinib (IC 50 ) sensitization of A549 and H460 cell lines comparing co-inhibition of AXL + SRC using siAXL or AXLi or CRISPR-knockout methods in an MTT assay (72 h). p -values are denoted by * ( p < 0.05), ** ( p < 0.01), *** ( p < 0.001) or **** ( p < 0.0001).

Article Snippet: Dasatinib and AXL inhibitor, SGI-7079, were purchased from Selleck chemicals (Houston, TX, USA).

Techniques: In Vitro, Inhibition, MTT Assay, CRISPR, Knock-Out

Journal: Cancers

Article Title: Synergistic Inhibition of Drug Resistant KRAS Mutant Non-Small Cell Lung Cancer by Co-Targeting AXL and SRC

doi: 10.3390/cancers17030490

Figure Lengend Snippet: Synergistic AXL and SRC therapy in vivo. ( a ) Tumor growth reduction in A549 xenografts treated with Dasatinib or combination of AXLi and Dasatinib ( b ) Corresponding graph showing tumor growth inhibition (%). ( c ) Tumor volume reduction plot for excised tumors. ( d ) Western blot image for DDR2, SHP2 and SRC activity in tumor lysates at the end of study. p -values are denoted by * ( p < 0.05), ** ( p < 0.01), *** ( p < 0.001) or **** ( p < 0.0001).

Article Snippet: Dasatinib and AXL inhibitor, SGI-7079, were purchased from Selleck chemicals (Houston, TX, USA).

Techniques: In Vivo, Inhibition, Western Blot, Activity Assay

Journal: Oncotarget

Article Title: Axl inhibition induces the antitumor immune response which can be further potentiated by PD-1 blockade in the mouse cancer models

doi: 10.18632/oncotarget.21125

Figure Lengend Snippet: ( A ) ID8 cells were treated with vehicle or R428 at indicated doses before performing proliferation, migration and invasion assays. ( B ) ID8 cells were transfected with mock, control siNC or siRNA specific for Axl (siAxl-2) and subsequently subjected to proliferation, migration and invasion assays. Proliferation and invasion was plotted as a percentage of growth relative to vehicle-treated cells with migration plotted as a percentage relative to zero time point of each treated cells. The experiments were performed thrice with 3 technical replicates, and data are expressed as mean ± SEM, ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, one-way ANOVA followed by Tukey's multiple comparisons test.

Article Snippet: Axl tyrosine kinase inhibitors R428 (BGB324) and SGI-7079 were purchased from Selleck Inc. R428 and SGI-7079 specificities for Axl have been previously evaluated [ , ].

Techniques: Migration, Transfection, Control

Journal: Oncotarget

Article Title: Axl inhibition induces the antitumor immune response which can be further potentiated by PD-1 blockade in the mouse cancer models

doi: 10.18632/oncotarget.21125

Figure Lengend Snippet: ( A ) Schematic regimen for testing the in vivo optimal dose of R428 (top) or R428 treatment in mice depleted of lymphocyte subsets (bottom). ( B ) C57BL/6 mice (6 per group) bearing 10-day-established ID8 tumors were treated with control vehicle or R428 at the indicated dose for two weeks and their overall survival was evaluated. ( C ) Nude mice (6 per group) bearing 10-day-established ID8 tumors were treated with R428 at the indicated dose for two weeks and their overall survival was evaluated. ( D ) C57BL/6 mice (6 per group) bearing 10-day-established ID8 tumors with lymphocyte subset depletion were treated with R428 at the indicated dose for two weeks and their overall survival was evaluated. Data are representative of two (B and D) or three (C) independent experiments, ** p < 0.01, *** p < 0.001, log-rank test (B–D).

Article Snippet: Axl tyrosine kinase inhibitors R428 (BGB324) and SGI-7079 were purchased from Selleck Inc. R428 and SGI-7079 specificities for Axl have been previously evaluated [ , ].

Techniques: In Vivo, Control

Journal: Oncotarget

Article Title: Axl inhibition induces the antitumor immune response which can be further potentiated by PD-1 blockade in the mouse cancer models

doi: 10.18632/oncotarget.21125

Figure Lengend Snippet: C57BL/6 mice (3 per group) bearing 10-day-established ID8 tumors were treated with control vehicle or R428 for 5 days and then TILs and immune-associated genes within tumors were analyzed by FACS and qPCR respectively. ( A ) The representative FACS plots, the percentage and absolute number of CD45 + TILs in tumors from vehicle or R428 treated mice. ( B ) The percentage and absolute number of CD4 + FoxP3 – T cells, CD8 + T cells, CD3 + DX5 + NK cells, CD11c + MHCII + cDCs, CD11b + F4/80 + Ly-6G – monocytes/macrophages (Mono/Mφ), CD11b + F4/80 – Ly-6G + granulocytes and CD4 + FoxP3 + Treg cells within TILs. ( C ) The percentages and representative FACS plots of CD69 + and Ki-67 + cells among tumor-infiltrating CD4 + T cells and CD8 + T cells. ( D ) The percentages and representative FACS plots of IFN-γ + cells among tumor-infiltrating CD4 + T cells and CD8 + T cells in response to phorbol myristyl acetate (PMA)/Ionomycin stimulation. ( E ) The expression of genes associated with type-1 T cell or suppressive cells recruitment and functionality. Mean values of messenger RNA levels in control group were set to 1. Data are derived from three tumors per treatment and representative of two independent experiments, * p < 0.05, ** p < 0.01, *** p < 0.001, two-tailed student t test.

Article Snippet: Axl tyrosine kinase inhibitors R428 (BGB324) and SGI-7079 were purchased from Selleck Inc. R428 and SGI-7079 specificities for Axl have been previously evaluated [ , ].

Techniques: Control, Expressing, Derivative Assay, Two Tailed Test

Journal: Oncotarget

Article Title: Axl inhibition induces the antitumor immune response which can be further potentiated by PD-1 blockade in the mouse cancer models

doi: 10.18632/oncotarget.21125

Figure Lengend Snippet: C57BL/6 mice (3 per group) bearing 10-day-established ID8 tumors were treated with control vehicle or R428 for 5 days and tumor-infiltrating cDCs were analyzed by FACS. ( A ) Representative FACS plots for CD103 versus CD11b within CD11c + MHCII + cDC gate, and percentage (within CD11c + MHCII + cells) and absolute number of CD103 + CD11c + MHCII + cDCs. ( B ) Representative FACS plots for CD86 versus CD40 within CD11b + or CD103 + CD11c + MHCII + gate and percentage of CD86 + CD40 + within CD11b + or CD103 + CD11c + MHCII + cDCs. ( C ) Representative FACS plots for CD11c versus IL-12p40 within CD11b + or CD103 + CD11c + MHCII + gate and percentage of IL-12p40 + within CD11b + or CD103 + CD11c + MHCII + cDCs. Data are representative of two independent experiments, * p < 0.05, ** p < 0.01, two-tailed student t test.

Article Snippet: Axl tyrosine kinase inhibitors R428 (BGB324) and SGI-7079 were purchased from Selleck Inc. R428 and SGI-7079 specificities for Axl have been previously evaluated [ , ].

Techniques: Control, Two Tailed Test

Journal: Oncotarget

Article Title: Axl inhibition induces the antitumor immune response which can be further potentiated by PD-1 blockade in the mouse cancer models

doi: 10.18632/oncotarget.21125

Figure Lengend Snippet: ( A – B ) C57BL/6 mice (3 per group) bearing 10-day-established ID8 tumors were treated with control vehicle, R428 or SGI-7079 for 5 days and tumor cells and tumor-infiltrating T cells were analyzed by FACS. (A) The representative FACS plots (left) and statistical results (right) of PD-1 expression on tumor-infiltrating CD4 + T cells and CD8 + T cells. (B) The representative FACS plots (left) and statistical results (right) of PD-L1 and MHC-I expression on tumor cells. ( C ) Schematic regimen for combined Axl inhibition (R428 or SGI-7079 treatment) and PD-1 blockade in ID8 tumor-bearing mice. ( D ) C57BL/6 mice (6 per group) bearing 10-day-established ID8 tumors were treated with either single or combined R428/SGI-7079 and anti-PD-1 mAb for two weeks and their overall survival was evaluated. ( E ) Ninety days after first tumor challenge, long-term survivors (LTS) were rechallenged with ID8 (i.p.) or unrelated TC1 (s.c.) tumor cells and their tumor growth was evaluated. Naïve mice with same challenge were used as controls. Data are representative of two (A and B) or three (D) independent experiments, ** p < 0.01, *** p < 0.001, one-way ANOVA followed by Tukey's multiple comparisons test (A and B) or log-rank test (D).

Article Snippet: Axl tyrosine kinase inhibitors R428 (BGB324) and SGI-7079 were purchased from Selleck Inc. R428 and SGI-7079 specificities for Axl have been previously evaluated [ , ].

Techniques: Control, Expressing, Inhibition