Fig 1: Concentration-dependent inhibition of EF1a-1 processing in cancer cells. A, A549 lung cancer cells were treated for 24 hours with the indicated concentrations (mol/L) of M8891 and analyzed for the presence of MetEF1a-1 by simple western blotting. Tubulin served as loading control. A virtual blot-like image generated by the Protein Simple Compass software is shown. B, A549 lung cancer cell line, HT1080 fibrosarcoma cell line, and U87MG glioblastoma cancer cell line were treated with the indicated concentrations of M8891 for 24 hours and analyzed by flow cytometry using MetEF1a-1 antibody. C, Genome-wide CRISPR screen to identify modifiers of pharmacologic MetAP2 inhibition. CRISPR screens were performed with A549, HT1080, and U87MG in the presence of IC40 concentrations of MetAP2 inhibitors, TNP-470 or M8891. normZ scores from both CRISPR screens were plotted (y axis: TNP-470, x axis: M8891) per cell line. Selected genes (MetAP1, MetAP2, CDKN1A, and TP53) are highlighted. Abbreviations: CRISPR, clustered regularly interspaced short palindromic repeats; EF1a-1, elongation factor 1-alpha 1; IC40, 40% inhibitory concentration; MetAP2, methionine aminopeptidase 2; MetEF1a-1, methionine-containing N-terminus of elongation factor 1-alpha 1; normZ, normalized gene-level z-score.
Fig 2: Antiangiogenic, antitumoral activities, and PK/PD relationship of M8891. A, Treatment scheme to assess blood vessel formation in VEGFR2-luc mice implanted with Matrigel. One day after implantation, mice were treated with M8891 (5, 10, 20 mg/kg, daily, per os) or anti-VEGFR antibody B20 (20 mg/kg, twice weekly, intravenously). After 14 days of treatment mice were treated with luciferin and Matrigel plugs were removed, and bioluminescence was quantified. Representative images for Matrigel plugs from vehicle- and M8891-treated mice (10 mg/kg dose) are shown. B, Quantification of bioluminescence from different treatment groups from the study outlined in A displayed as a bar diagram ± standard error of the mean. Statistically significant reductions of bioluminescence in comparison with vehicle-treatment group are indicated by *. C, Tumor growth curves of ccRCC model Caki-1 treated with vehicle or different doses of M8891 (10, 25, 50 mg/kg, twice daily, per os). Statistically significant differences versus vehicle-treated control group are indicated (***, P < 0.0001; *, P < 0.05). D, Relationship of PK to PD effect on inhibition of N-terminal processing of EF1a-1 by M8891 administered to Caki-1 tumor-bearing mice. Plasma levels of M8891 and formation of unprocessed MetEF1a-1 in different treatment groups of M8891 (10, 25, or 100 mg/kg, single administration, per os) is shown as a function of time. In tumors from vehicle-treated animals sampled at 24 hours, MetEF1a1 was undetectable. E, Tumor growth curves of gastric cancer PDX model GXF-1172 treated with vehicle or M8891 (150 mg/kg, twice daily, per os). Mice in the vehicle-treated group were sacrificed on day 28 due to reaching a mean tumor volume >800 mm³. At treatment end with M8891 (day 80), 7 of 10 tumor-bearing mice were still alive, three mice were removed due to tumor progression on days 35 (two mice) and 49 (one mouse). F, Tumor growth curves of colon cancer PDX model CXF-1783 treated with vehicle or M8891 (150 mg/kg, twice daily, per os). Selected mice in the vehicle-treated group were sacrificed due to reaching the tumor growth limit on the following days: 24 (n = 2), 31 (n = 3), 35 (n = 2), 42 (n = 1), and 49 (n = 3). On the last treatment day with M8891 (day 80), 6 of 10 tumor-bearing mice were still alive and four were removed due to tumor progression on days 49 (n = 2), 56, and 59. Abbreviations: EF1a-1, elongation factor 1-alpha 1; NFG, Matrigel without growth factors; MetEF1a-1, methionine-containing N-terminus of elongation factor 1-alpha 1; PD, pharmacodynamics; PK, pharmacokinetics; SEM, standard error of the mean; VEGFR, VEGF receptor.
Supplier Page from Abcam for Recombinant Human eEF1A1/EF-Tu protein (denatured) (Tag Free)