<p>Figure S1. Kinase interaction map for AMG 337; Figure S2. In a large unbiased cancer cell line viability screen only MET-amplified cell lines were sensitive to treatment with an analogue of AMG 337 (Compound 5); Figure S3: AMG 337 inhibits the phosphorylation of MET and but not its downstream effectors in MET-amplified, KRAS mutant NSCLC cell line NCI-H1573; Figure S4. Cell lines harboring MET FISH scores >3 exhibited sensitivity to AMG 337. MET FISH analysis was performed on a subset of cancer cell lines exhibiting elevated MET gene copy number; Figure S5. Selective inhibition of MET exhibits partial effects on the viability of U-87 MG glioblastoma cells harboring an HGF/MET autocrine loop; Figure S6. Increases in MET gene number correlate with high levels of total MET protein; Figure S7. AMG 337 inhibits Gab-1 phosphorylation in a concentration dependent manner in the TPR-MET mouse tumor model.</p>
Supplementary Figure 2 from Identification of a Novel Recepteur d'Origine Nantais/c-Met Small-Molecule Kinase Inhibitor with Antitumor Activity <i>In vivo</i>
Supplementary Figure 1 from Identification of a Novel Recepteur d'Origine Nantais/c-Met Small-Molecule Kinase Inhibitor with Antitumor Activity <i>In vivo</i>
4976 c-Met is a receptor tyrosine kinase that signals through its ligand, hepatocyte growth factor (HGF), to mediate multiple biological activities including proliferation, survival, motility and migration. In cancer, HGF/c-Met signaling is often deregulated. Overexpression of c-Met occurs frequently in numerous types of tumors. Gene amplification and activating mutations leading to constitutive receptor activation have also been described in patient tumors. In addition, altered expression of this axis in several mouse models has implicated c-Met in tumorigenesis. Therefore, targeting c-Met has promise as an approach to treating patients with tumors harboring activated c-Met. AMG 458 was identified as a selective, orally bioavailable, ATP-competitive small molecule c-Met inhibitor. AMG 458 potently inhibited c-Met enzymatic activity with an IC50 of 2 nM and blocked c-Met phosphorylation in PC3 cells with an IC50 of 60 nM. AMG 458 inhibited signaling in a dose-dependent manner in vitro in NIH-3T3 cells stably expressing TPR-Met, a constitutively active form of c-Met. Consistent with its activity in vitro, AMG 458 significantly inhibited HGF-mediated c-Met phosphorylation in the mouse liver at 30 mg/kg PO (p
Abstract Homozygous deletion of chr9p21, containing genes CDKN2A and MTAP, occurs in about 15% of cancers. MTAP loss, a key enzyme in the methionine and adenine salvage pathways, leads to the accumulation of its substrate MTA. MTA competes with the methyl donor SAM for binding to type II arginine methyltransferase PRMT5, placing PRMT5 in a hypomorphic state and vulnerable to further PRMT5 inhibition. MTA-cooperative PRMT5 inhibitors are an emerging class of therapeutics targeting MTAP null tumors. AMG 193, currently in Phase 1 trials, and its representative analog AM-9747 have broad spectrum activity in MTAP null tumor models across hematologic and solid tumor indications. Mechanistically, AM-9747 induces DNA damage, shown by increased phosphorylated H2AX, resulting in cell cycle arrest and senescence. Exploration of clinically relevant therapeutic combinations with standard of care (SOC) chemotherapies or targeted agents that could potentiate this DNA damage or target orthogonal pathways is a rational therapeutic approach. Here, we evaluated AM-9747 in combination with SOC chemotherapies and assessed synergy using the Chou-Talalay Method to generate Combination Index (CI) scores. SOC agents representing a variety of mechanisms of action (paclitaxel, carboplatin, gemcitabine, pemetrexed, irinotecan, 5-FU) were evaluated and results ranged from strongly synergistic (CI<0.3) to additive (CI=1) in a panel of non-small cell lung carcinoma (NSCLC) and pancreatic cancer cell lines. The SOC combinations augmented AM-9747 induced DNA damage resulting in increased cell cycle arrest and nuclear count assays confirmed the synergistic antiproliferative effects of the combinations. Combinations of AM-9747 and one strongly synergistic agent (carboplatin) and one moderately synergistic agent (paclitaxel) were evaluated in H292 NSCLC xenografts. Significant reductions in tumor burden compared to single agent treatments were observed with AM-9747 + paclitaxel (67% TGI) and AM-9747 + carboplatin (82% TGI). Additionally, roughly 2-3% of NSCLC tumors harbor both MTAP loss and a KRAS G12C mutation. To target both pathways we combined the FDA-approved KRAS G12C inhibitor sotorasib with AM-9747 and observed synergy (CI<0.6) in MIAPACA2 cells. Nuclear counts confirmed a greater cell growth inhibition in the combination and immunoblots confirmed on target inhibition of both pathways. Combination treatment of AM-9747 and sotorasib in LU99 NSCLC xenografts resulted in significant tumor regression compared to each single agent with 10/10 mice tumor free. This combination is being tested in additional MTAP null KRAS G12C mutant models. Overall, our data suggests that combining MTA-cooperative inhibitor AM-9747 with SOC or clinically relevant targeted agents is a compelling therapeutic strategy for the treatment of MTAP null cancers. Citation Format: Katherine Slemmons, Brian Belmontes, Siyuan Liu, Jodi Moriguchi, Antonia Policheni, Paul E. Hughes. The MTA-cooperative PRMT5 inhibitor AM-9747 exhibits robust antitumor activity in combination with clinically relevant chemotherapies and targeted agents in MTAP null tumor models. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6246.
Abstract Bevacizumab is a monoclonal antibody targeting VEGF that is clinically approved for the treatment of multiple indications in oncology, including colorectal, lung and renal cancer as well as glioblastoma. Clinical activity has also been demonstrated in breast and ovarian cancer as well, however, the time periods of benefit for all of these indications is in months (1-5). Multiple pathways are known to participate in the process of angiogenesis, and the limited effects of bevacizumab may represent the maximum activity of targeting just one of them. The purpose of the present study was to examine the effect of simultaneously targeting multiple angiogenic axes including angiopoietins 1 and 2, Dll4 and VEGF. Trebananib is a recombinant peptide-Fc fusion protein that selectively blocks the Tie2 interactions of both Ang1 and Ang2. Trebananib, as well as a recombinant antibody to Dll4 were produced at Amgen, Inc., Thousand Oaks, CA. Bevacizumab was purchased from a local pharmacy. These reagents were employed in the combination treatment of three different preclinical murine tumor models (U87, glioblastoma; MiaPaca and BxPC3, pancreatic carcinoma), one of which (BxPC3) was specifically selected for its relative resistance to VEGF/KDR inhibition. Harlan Athymic nu/nu female mice at 6-7 weeks of age were injected subcutaneously on the right flank with 5 x 106 tumor cells in the presence of Matrigel and treated when the tumors were approximately 300 mm3. The results, as supported by histomorphometric analysis (percent blood vessel area, viable tumor burden), demonstrate that under conditions of optimal biological dosing, the addition of trebananib to a two-way combination of bevacizumab and anti-Dll4 resulted in a statistically significant enhancement of therapeutic effect for the three-way combination, an observation made consistently over 3 different tumor models and 3 different dose levels of bevacizumab and anti-Dll4 (p = or < 0.0024; RMANOVA). Furthermore, the therapeutic effect in all three models approximated prolonged stasis, the ultimate theoretical goal of antiangiogenesis in the treatment of cancer. 1. NEJM, vol. 350, p. 2335, 2004. 2. NEJM, vol. 355, p. 2542, 2006. 3. Lancet, vol. 370, p. 2103, 2007. 4. NEJM, vol. 357, p. 2666, 2007. 5. J. Clin. Oncol., vol. 30, p. 2039, 2012. Citation Format: Bethany Mattson, Jodi Moriguchi, H. Toni Jun, Angela Coxon, Dave Cordover, Steve Kaufman, Jon Oliner, Charlie Starnes. Triple combination of bevacizumab, anti-DLL4 (delta like ligand 4) and trebananib gives enhanced therapeutic effects in three xenograft tumor models. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2991. doi:10.1158/1538-7445.AM2014-2991
