copanlisib pi3k inhibitor  (MedChemExpress)

Journal: International journal of molecular sciences

Article Title: Protein Tyrosine Phosphatase Non-Receptor 11 ( PTPN11 /Shp2) as a Driver Oncogene and a Novel Therapeutic Target in Non-Small Cell Lung Cancer (NSCLC).

doi: 10.3390/ijms241310545

Figure Lengend Snippet: Figure 3. PTPN11 mutations result in elevated SHP2-phosphatse activity and activate MAPK and PI3K pathway signalling. Serum-starved cells treated with 100 ng/mL EGF for 5 min were lysed, and SHP2 was immunoprecipitated from whole cell lysates. Immunoprecipitate was used to de- termine phosphatase activity, as described in the materials and methods. (A) SHP2-phosphatse activity in H661 (PTPN11-mutated) compared to H1703 (PTPN11-WT), Calu-3 (PTPN11-WT), and H157 (PTPN11-WT, KRAS-mutated). Shp2-phosphatase activity in H1701 (B) and H1299 (C) cells transduced with indicated PTPN11 mutations. Data are represented as mean ± s.e.m. (n = 3). (D) NCI-H1703 and NCI-H157 cells were transduced with wildtype or mutated PTPN11: a serum starved and stimulated with an epidermal growth factor (100 ng/mL) for 5 min. p-ERK1/2—phospho- ERK 1/2; t-ERK1/2—total ERK 1/2; p-AKT ser 473—phospho-AKT (phosphorylated at serine 473); t-AKT—total AKT. Blots are representative of 3 independent experiments. (NS = No Significance, * p < 0.05, ** p < 0.01, *** p <0.001)

Article Snippet: The Shp2 inhibitor (SHPi) (SHP099), PI3K inhibitor copanlisib (cop) and the MEK1/2 inhibitor refametinib (ref) were obtained from Selleckchem and stocks (10 mM SHPi, 10 mM refametinib, 5 mM copanlisib;) were prepared in 100% DMSO, 100% DMSO and 100% DMSO with 10 mM TFA, respectively.

Techniques: Activity Assay, Immunoprecipitation, Transduction

Journal: International journal of molecular sciences

Article Title: Protein Tyrosine Phosphatase Non-Receptor 11 ( PTPN11 /Shp2) as a Driver Oncogene and a Novel Therapeutic Target in Non-Small Cell Lung Cancer (NSCLC).

doi: 10.3390/ijms241310545

Figure Lengend Snippet: Figure 5. SHP2 inhibitor (SHPi) improves the response to MAPK and PI3K pathway targeting therapies. H661 and H1703 cells plated and after 24 h were treated with SHPi (10 µM) daily or Cop (20 nM) or Ref (10 µM) alone or in combination with SHPi. 72 h following treatment, an Alamar Blue cell viability assay was performed (A). Data (n = 3 independent experiments) are expressed as mean ± SEM. Statistical significance was determined using one-way ANOVA correcting for multiple comparisons using Tukey’s test and reporting adjusted p values. (* p < 0.05, ** p < 0.01).

Article Snippet: The Shp2 inhibitor (SHPi) (SHP099), PI3K inhibitor copanlisib (cop) and the MEK1/2 inhibitor refametinib (ref) were obtained from Selleckchem and stocks (10 mM SHPi, 10 mM refametinib, 5 mM copanlisib;) were prepared in 100% DMSO, 100% DMSO and 100% DMSO with 10 mM TFA, respectively.

Techniques: Viability Assay

Journal: International journal of molecular sciences

Article Title: Protein Tyrosine Phosphatase Non-Receptor 11 ( PTPN11 /Shp2) as a Driver Oncogene and a Novel Therapeutic Target in Non-Small Cell Lung Cancer (NSCLC).

doi: 10.3390/ijms241310545

Figure Lengend Snippet: Figure 6. SHPi prevents feedback activation of MAPK and PI3K pathways following targeted treatments. H661 and H1703 cells seeded at a density of 2 × 105 cells per well and were treated 24 h later with SHPi (10 µM) daily or Cop (20 nM) or Ref (10 µM) alone or in combination with SHPi for 72 h. The expression of pAKT and pERK was analysed using Western blotting (representative images displayed) (A,B), and densitometry was quantified using either ImageJ or ImageLab software 5.2.1. (The volume intensity of the band relative to actin was matched to the total controls and displayed relative to the vehicle control). H661 Results are expressed as mean ± SEM (n = 4) (C,D), and H1703 results are expressed as mean ± SEM (n = 3) (E,F). Statistical significance was determined using one- way ANOVA correcting for multiple comparisons using Tukey’s test and reporting adjusted p values. (* p <0.05, **** p < 0.0001).

Article Snippet: The Shp2 inhibitor (SHPi) (SHP099), PI3K inhibitor copanlisib (cop) and the MEK1/2 inhibitor refametinib (ref) were obtained from Selleckchem and stocks (10 mM SHPi, 10 mM refametinib, 5 mM copanlisib;) were prepared in 100% DMSO, 100% DMSO and 100% DMSO with 10 mM TFA, respectively.

Techniques: Activation Assay, Expressing, Western Blot, Software, Control

Journal: International journal of molecular sciences

Article Title: Protein Tyrosine Phosphatase Non-Receptor 11 ( PTPN11 /Shp2) as a Driver Oncogene and a Novel Therapeutic Target in Non-Small Cell Lung Cancer (NSCLC).

doi: 10.3390/ijms241310545

Figure Lengend Snippet: Figure 7. PTPN11 and PI3K targeting treatments do not alter tumour formation or invasion in a chick embryo xenograft model. 2 × 106 H661 cells were implanted into the CAM according to the assay schedule (A) on day 7 of embryonic development. Following 72 h of tumour establishment, developing tumours were treated in situ. SHPi (10 µM) treatments were added daily, and Cop (20 nM) treatments were added once on day 10. On day 14, tumour visibility was noted as either visible (VT) or non-visible (NVT). Statistical significance was determined using Fisher’s exact test, and no statistical significance was found (B) On day 14, xenografts were excised with the silicon ring, formalin-fixed, and stained for H&E (C,D). Areas of tumour were denoted by black arrows, and CAM areas were demonstrated by red arrows and matrigel was denoted by grey arrows. Images were collected an EVOS m5000 microscope with EVOS imaging software at 4× and 20× magnification (D).

Article Snippet: The Shp2 inhibitor (SHPi) (SHP099), PI3K inhibitor copanlisib (cop) and the MEK1/2 inhibitor refametinib (ref) were obtained from Selleckchem and stocks (10 mM SHPi, 10 mM refametinib, 5 mM copanlisib;) were prepared in 100% DMSO, 100% DMSO and 100% DMSO with 10 mM TFA, respectively.

Techniques: In Situ, Staining, Microscopy, Imaging, Software

Journal: Anti-Cancer Drugs

Article Title: RIDR-PI-103, ROS-activated prodrug PI3K inhibitor inhibits cell growth and impairs the PI3K/Akt pathway in BRAF and MEK inhibitor-resistant BRAF-mutant melanoma cells

doi: 10.1097/CAD.0000000000001500

Figure Lengend Snippet: Summary of the efficacy of RIDR-PI-103 in BRAF and MEK inhibitor-resistant BRAF-mutant melanoma. RIDR-PI-103 (a prodrug of PI-103) is activated in a high ROS environment in TDR cells and releases biologically active PI-103. PI-103 is a pan-PI3K inhibitor and inhibits the activation of Akt and S6. RIDR-PI-103 inhibits cell proliferation, growth, and migration in TDR cells. Figure generated using biorender.com. ANOVA, analysis of variance; RIDR, ROS-induced drug release; ROS, reactive oxygen species.

Article Snippet: Only five PI3K inhibitors – Copanlisib (Aliqopa), Idelalisib (Zydelig), Alpelisib (PIQRAY), Duvelisib (COPIKTRA), and Umbralisib (Ukoniq) have been FDA approved to date [ ].

Techniques: Mutagenesis, Activation Assay, Migration, Generated