Supplemental Table 1. Biochemical and Cellular Potency and Selectivity of PF

Similar documents
SUPPLEMENTARY INFORMATION

(A) RT-PCR for components of the Shh/Gli pathway in normal fetus cell (MRC-5) and a

Supplementary Figure 1. Basal level EGFR across a panel of ESCC lines. Immunoblots demonstrate the expression of phosphorylated and total EGFR as

Supplementary Materials. for Garmy-Susini, et al, Integrin 4 1 signaling is required for lymphangiogenesis and tumor metastasis

A263 A352 A204. Pan CK. pstat STAT3 pstat3 STAT3 pstat3. Columns Columns 1-6 Positive control. Omentum. Rectosigmoid A195.

Supplemental Figure 1. Western blot analysis indicated that MIF was detected in the fractions of

Supplementary Materials for

Supplemental Table S1: Inhibition of HDAC class I and class II family by CUDC-101 (IC50 in nm)

Supplementary Figure (OH) 22 nanoparticles did not affect cell viability and apoposis. MDA-MB-231, MCF-7, MCF-10A and BT549 cells were

Supplementary information

(a) Schematic diagram of the FS mutation of UVRAG in exon 8 containing the highly instable

SUPPLEMENTARY INFORMATION

Supplementary Table 1. Antibodies and dilutions used in the immunohistochemical study

(A) PCR primers (arrows) designed to distinguish wild type (P1+P2), targeted (P1+P2) and excised (P1+P3)14-

ALM301: Allosteric Isoform selective Akt inhibitor

(PDGF), 9 ( -2 (FGF-2), SMO

Supplementary Information and Figure legends

A549 and A549-fLuc cells were maintained in high glucose Dulbecco modified

Supplemental Figure 1. Isolation and characterization of CD133+ neurosphere-like

(A) Cells grown in monolayer were fixed and stained for surfactant protein-c (SPC,

A Hepatocyte Growth Factor Receptor (Met) Insulin Receptor hybrid governs hepatic glucose metabolism SUPPLEMENTARY FIGURES, LEGENDS AND METHODS

Supplementary Figure 1.TRIM33 binds β-catenin in the nucleus. a & b, Co-IP of endogenous TRIM33 with β-catenin in HT-29 cells (a) and HEK 293T cells

Supplemental Data. Prolonged Rapamycin Treatment Inhibits mtorc2 Assembly and Akt/PKB. Supplemental Experimental Procedures

Supplementary Figure 1. Characterization of NMuMG-ErbB2 and NIC breast cancer cells expressing shrnas targeting LPP. NMuMG-ErbB2 cells (a) and NIC

RXDX-101 & RXDX-102. Justin Gainor, MD February 20 th, 2014

Impact of hyper-o-glcnacylation on apoptosis and NF-κB activity SUPPLEMENTARY METHODS

Supplementary Figure S1. Supplementary Figure S1. Chemical structure of CH

Supplementary Information. Targeting BRK-positive Breast Cancer with Small

Supplementary Figure 1. EC-specific Deletion of Snail1 Does Not Affect EC Apoptosis. (a,b) Cryo-sections of WT (a) and Snail1 LOF (b) embryos at

Antitumor Activity of CUDC-305, a Novel Oral HSP90 Inhibitor, in Solid and Hematological Tumor Xenograft Models

Supplementary Figure 1. Identification of tumorous sphere-forming CSCs and CAF feeder cells. The LEAP (Laser-Enabled Analysis and Processing)

Supplemental figure 1. PDGFRα is expressed dominantly by stromal cells surrounding mammary ducts and alveoli. A) IHC staining of PDGFRα in

Immunostaining was performed on tumor biopsy samples arranged in a tissue-microarray format or on

Type of file: PDF Size of file: 0 KB Title of file for HTML: Supplementary Information Description: Supplementary Figures

Nature Neuroscience: doi: /nn Supplementary Figure 1

Supplemental Figure 1. Intracranial transduction of a modified ptomo lentiviral vector in the mouse

Supplementary Information

Co-Development of DELFIA Technology. - Assessment of DELFIA technology in drug biodistribution in small animals

Supplementary Figure 1: si-craf but not si-braf sensitizes tumor cells to radiation.

To determine the effect of over-expression and/or ligand activation of. PPAR / on cell cycle, cell lines were cultured as described above until ~80%

Mechanisms of resistance to JAK inhibitors. L. Knoops

Incorporation of photo-caged lysine (pc-lys) at K273 of human LCK allows specific control of the enzyme activity.

Figure S1. Generation of inducible PTEN deficient mice and the BMMCs (A) B6.129 Pten loxp/loxp mice were mated with B6.

Supplementary Figure 1. Characterization of ALDH-positive cell population in MCF-7 cells. (a) Expression level of stem cell markers in MCF-7 cells or

Nieuwe targets en cfdna

Supplementary Figure 1. A. Bar graph representing the expression levels of the 19 indicated genes in the microarrays analyses comparing human lung

Supplementary Figure 1: Expression of NFAT proteins in Nfat2-deleted B cells (a+b) Protein expression of NFAT2 (a) and NFAT1 (b) in isolated splenic

Supplementary Information POLO-LIKE KINASE 1 FACILITATES LOSS OF PTEN-INDUCED PROSTATE CANCER FORMATION

Figure S1. PMVs from THP-1 cells expose phosphatidylserine and carry actin. A) Flow

(a) Significant biological processes (upper panel) and disease biomarkers (lower panel)

SUPPLEMENTARY MATERIAL

Fang et al. NMuMG. PyVmT unstained Anti-CCR2-PE MDA-MB MCF MCF10A

Supplementary Figure 1

TITLE: Notch in Pathological Angiogenesis and Lymphangiogenesis

Osamu Tetsu, MD, PhD Associate Professor Department of Otolaryngology-Head and Neck Surgery School of Medicine, University of California, San

SUPPLEMENTARY FIGURES

hexahistidine tagged GRP78 devoid of the KDEL motif (GRP78-His) on SDS-PAGE. This

Supplemental Information. Otic Mesenchyme Cells Regulate. Spiral Ganglion Axon Fasciculation. through a Pou3f4/EphA4 Signaling Pathway

Supplementary Figure 1 CD4 + T cells from PKC-θ null mice are defective in NF-κB activation during T cell receptor signaling. CD4 + T cells were

Supplementary Figures

IntelliGENSM. Integrated Oncology is making next generation sequencing faster and more accessible to the oncology community.

B16-F10 (Mus musculus skin melanoma), NCI-H460 (human non-small cell lung cancer

mtor Inhibition Specifically Sensitizes Colorectal Cancers with KRAS or BRAF Mutations to BCL-2/BCL-

BCR-ABL uncouples canonical JAK2-STAT5 signaling in chronic myeloid

Supplementary Table 1. List of primers used in this study

Inhibitors of Methionine Aminopeptidase-2 2 in the Treatment of Non-Hodgkin

SHREE ET AL, SUPPLEMENTAL MATERIALS. (A) Workflow for tumor cell line derivation and orthotopic implantation.

Supplementary Figure 1. PD-L1 is glycosylated in cancer cells. (a) Western blot analysis of PD-L1 in breast cancer cells. (b) Western blot analysis

c-abl PDGFRα PDGFRβ VEGFR-1 VEGFR-2 FLT-3 c-fms c-kit Ref. 2 (ND) 9.4 (ND) Supplementary Table 2. Chemical structure of the RTKIs used in this study.

Pharmacologic inhibition of histone demethylation as a therapy for pediatric brainstem glioma

p = formed with HCI-001 p = Relative # of blood vessels that formed with HCI-002 Control Bevacizumab + 17AAG Bevacizumab 17AAG

CD14 + S100A9 + Monocytic Myeloid-Derived Suppressor Cells and Their Clinical Relevance in Non-Small Cell Lung Cancer

SUPPLEMENTARY INFORMATION

Targeting the Molecular Chaperone Hsp90 in Cancer: What Does the Biology Tell Us? Len Neckers TAT 2011

Table S1. New colony formation 7 days after stimulation with doxo and VCR in JURKAT cells

RayBio KinaseSTAR TM Akt Activity Assay Kit

Scavenger receptor class B type I regulates cellular cholesterol metabolism and cell signaling associated with breast cancer development.

FGL2 A new biomarker for cancer in a simple blood test

Effective Targeting of Quiescent Chronic Myelogenous

PREPARED FOR: U.S. Army Medical Research and Materiel Command Fort Detrick, Maryland

Clinical Policy: Cetuximab (Erbitux) Reference Number: PA.CP.PHAR.317

Supplementary Materials for

SUPPLEMENT. Materials and methods

VEGFR2-Mediated Vascular Dilation as a Mechanism of VEGF-Induced Anemia and Bone Marrow Cell Mobilization

PLX7486 Background Information October Candidate for CRUK Combinations Alliance

SUPPLEMENTARY FIGURES AND TABLE

2 nd line Therapy and Beyond NSCLC. Alan Sandler, M.D. Oregon Health & Science University

(A) Dose response curves of HMLE_shGFP (blue circle), HMLE_shEcad (red square),

HCC1937 is the HCC1937-pcDNA3 cell line, which was derived from a breast cancer with a mutation

SUPPLEMENTARY INFORMATION

BRaf V600E cooperates with Pten silencing to elicit metastatic melanoma (Nature Genetics Supplementary Information)

Supplemental Data. TGF-β-mediated mir-181a expression promotes breast cancer metastasis by targeting Bim.

Supplemental Information. Differential Effects of EGFL6 on Tumor. versus Wound Angiogenesis

Supplementary Figure 1: Characterisation of phospho-fgfr-y463 antibody. (A)

MECHANISMS OF 1ST GENERATION RESISTANCE. Byoung Chul Cho, M.D., Ph.D.

SUPPLEMENTARY INFORMATION. Supplementary Figures S1-S9. Supplementary Methods

Supplementary Figure 1

Postn MCM Smad2 fl/fl Postn MCM Smad3 fl/fl Postn MCM Smad2/3 fl/fl. Postn MCM. Tgfbr1/2 fl/fl TAC

Cancer therapeutics in the clinical setting. Need for alternate therapeutics. Target multiple cancer pathways. Circumvent the genetic mutations

Supporting Information

Transcription:

Supplemental Table 1. Biochemical and Cellular Potency and Selectivity of PF- 02341066

Assay IC 50 nm Selectivity Ratio d Biochemical Activity In Vitro c-met/hgfr enzyme (Ki, nm) a 4 NA Cellular Activity In Vitro c-met phosphorylation in human tumor cell lines (mean IC 50 ) b,c 11 NA c-met phosphorylation in mouse IMCD3 epithelial cells (IC 50 ) b 5 NA Phosphorylation of c-met WT in NIH3T3 cells (IC 50 ) b 13 NA Phosphorylation of c-met mutant V1092I in NIH3T3 cells (IC 50 ) b 19 NA Phosphorylation of c-met mutant H1094R in NIH3T3 cells (IC 50 ) b 2 0.1X Phosphorylation of c-met mutant Y1230C in NIH3T3 cells (IC 50 ) b 127 11X Phosphorylation of c-met mutant Y1235D in T47D cells (IC 50 ) b 92 8X Phosphorylation of c-met mutant M1250T in NIH3T3 cells (IC 50 ) b 15 NA Phosphorylation of c-met in NCI-H69 cells expressing c-met R988C variant (IC 50 ) b 13 NA Phosphorylation of c-met in HOP92 cells expressing c-met T1010I variant (IC 50 ) b 16 NA NPM-ALK phosphorylation in human Karpas299 lymphoma cells (IC 50 ) b 24 2X Cellular Activity Against Non-Target Kinases In Vitro MSP-stimulated RON phosphorylation in NIH-3T3-RON cells (mean IC 50 ) b 189 17X RON phosphorylation in RON-GYRB cells (mean IC 50 ) b 298 27X Gas6-stimulated Axl phosphorylation in NIH-3T3-Axl cells (mean IC 50 ) b 322 29X Ligand-stimulated Tie-2 phosphorylation in NIH-3T3-Tie-2/EGFR cells (mean IC 50 ) b 448 41X NGF-stimulated TrkA phosphorylation in Trk A-PAE cells (mean IC 50 ) b 580 53X BDNF-stimulated TrkB phosphorylation in Trk B-PAE cells (mean IC 50 ) b 399 36X BCR-Abl phosphorylation in BCR-Abl-BaF3 cells (mean IC 50 ) b,e 1159 >100X Insulin-stimulated insulin receptor phosphorylation in 293-IRK cells (mean IC 50 ) b 2887 >250X CD3-stimulated Lck-dependent Zap70 phosphorylation in Jurkat cells (mean IC 50 ) b 2741 >250X Gas6-stimulated Sky phosphorylation in NIH-3T3-Sky cells (mean IC 50 ) b >10000 ~1000X Definitions a Ki for c-met/hgfr enzyme inhibition determined by monitoring NADH oxidation coupled to ATP turnover. b IC 50 values were determined after exposure of various cell lines to several concentrations of PF-02341066 for 1 hour and measuring phosphorylation in cellular protein lysates by ELISA as described in Materials and Methods. IC 50 values were generated by curve fitting using a four-parameter analysis. c Mean IC 50 value derived from mean IC 50 value for c-met/hgfr phosphorylation across a panel of 7 human tumor cell lines (i.e., A549, MDA-MB-231, GTL-16, HT29, 786-O, Colo-205, A498). d Mean c cell c-met IC 50 was used to calculate the selectivity ratio for cell assays. Selectivity Index was calculated as IC 50, (c-met)/ic 50, (target). e BaF3 cells engineered to express human BCR-Abl were acquired from Dr. Brian Druker (Oregon Health and Science University, Portland). WT = Wild Type Antibody reagents and growth factors: Anti-phospho Zap70 was acquired from Zymed/Invitrogen (Carlsbad, CA). Anti-RON, FGFR1, PDGFRβ, Trk, Tie-2, and anti-phospho tyrosine (PY-20) were acquired from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-Axl and Sky antibodies and MSP, Gas6, NGF, EGF, and Insulin were acquired from R&D Systems (Minneapolis, MN). Anti-IRK was acquired from BD Pharmingen (San Diego, CA). Anti-VEGFR2 was acquired from Novus Biologicals (Littleton, CO). Anti-total and phospho ALK and anti-c- ABL were acquired from Cell Signaling Technologies (Boston, MA). BDGF was acquired from GibcoBRL/Invitrogen (Carlsbad, CA).

Supplemental Table 2. Effect of PF-02341066 on c-met -Dependent Phenotypes in Cancer Cells PF-02314066 Assay Concentration IC 50 (nm) a Tumor Cell Phenotypes Proliferation (MTT assay) of GTL-16 gastric carcinoma cells (mean IC 50 ) 9.7 Apoptosis (ccdna assay) of GTL-16 gastric carcinoma cells (mean IC 50 ) 8.4 HGF-stimulated NCI-H441 NSCLC cell Boyden Chamber migration (IC 50 ) 11 HGF-stimulated NCI-H441 NSCLC cell Boyden Chamber Matrigel invasion (IC 50 ) 6.1 HGF-stimulated MDCK cell colony scattering (mean IC 50 ) 16 Endothelial Cell Phenotypes HGF-stimulated HUVEC endothelial cell survival (MTT assay) (mean IC 50 ) 11 HGF-stimulated HUVEC cell Matrigel invasion (mean IC 50 ) 35 HMVEC endothelial cell tubulogenesis in fibrin gels (estimated IC 50 ) 80 a IC 50 values were generated by curve fitting using a four-parameter analysis. Definitions: NSCLC=non-small cell lung cancer; MDCK=Madin-Darby Canine Kidney; HUVEC=human umbilical vein endothelial cells; HMVEC=human microvascular endothelial cells

Supplemental Figure Legends Supplemental Figure 1. PF-02341066 (R)-3-[1-(2,6-Dichloro-3-fluoro-phenyl)- ethoxy]-5-(1-piperidin-4-yl-1h-pyrazol-4-yl)-pyridin-2-ylamine Supplemental Figure 2. PF-02341066 Inhibited HGF-stimulated NCI-H441Cell Migration and Invasion in vitro- NCI-H441 cells were treated with HGF (25 ng/ml) or designated concentrations of PF-02341066 and migration of cells across a membrane (Top Panel) and/or a layer of matrigel (Invasion--Lower Panel) to the lower well of a 2- chamber plate was assessed after 22 hours. Cells that invaded or migrated to the lower well of the plate were then fixed, nuclei were stained with DAPI, and the number of migrating or invading cells was determined utilzing ImagePro Plus software. Supplemental Figure 3. Dose-Dependent Induction of Apoptosis by PF-2341066 in Tumor Xenografts In Vivo. Immunohistochemical evaluation of activated caspase-3 expression was determined in GTL-16 (A) or U87MG (B) tumors at 50 mg/kg/day qd on study day 4. For each study, athymic mice bearing established GTL-16 or U87MG xenografts were orally administered vehicle or PF-2341066 at the indicated dose levels qd. At designated study days at 4 hours post-administration of PF-2341066, mice were humanely euthanized, tumors were resected and fixed in 10% neutral buffered formalin for 24 hours, and then placed in 70% ethanol. Fixed tumors were embedded in paraffin, cut into 4 mm sections, and immunostained for activated caspase-3 as described in Materials and Methods. Slides were visually assessed for expression and distribution of activated caspase-3 by light microscopy.

Supplemental Figure 1 NH N N Cl O N Supplemental Figure 2 F Cl NH 2 Supplemental Figure 3 A. B.

2024 © healthdocbox.com Privacy Policy | Terms of Service | Feedback