Tepotinib

Manufactured by Selleck Chemicals

Sourced in United States

Tepotinib is a laboratory reagent used for scientific research purposes. It is a small molecule that functions as a kinase inhibitor. The core function of Tepotinib is to inhibit the activity of the MET receptor tyrosine kinase.

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Lab products found in correlation

7 protocols using tepotinib

Apoptosis Detection Using Ramucirumab

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Ramucirumab, paclitaxel, and tepotinib were obtained from Selleck Chemicals (Houston, TX, USA). The annexin V-APC/propidium iodide (PI) apoptosis detection kit was obtained from Thermo Fisher Scientific (Waltham, MA, USA).

Sohn S.H., Sul H.J., Kim B.J, & Zang D.Y. (2024). Comparison of Tepotinib, Paclitaxel, or Ramucirumab Efficacy According to the Copy Number or Phosphorylation Status of the MET Gene: Doublet Treatment versus Single Agent Treatment. International Journal of Molecular Sciences, 25(3), 1769.

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Evaluating EGF, NRG1b, and HGF Signaling

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Recombinant human epidermal growth factor (EGF) and neuregulin 1b (NRG1b) were purchased from R&D Systems (Minneapolis, MN). Recombinant human hepatocyte growth factor (HGF) was purchased from Peprotech (Rocky Hill, NJ). Collagen was obtained from Advanced Biomatrix (Carlsbad, CA) and fibronectin was obtained from Sigma (St. Louis, MO). Neratinib, erlotinib, tepotinib were purchased from SelleckChem (Houston, TX) and prepared at stock concentrations in fresh 100% DMSO (Amresco) before final dilution into assay medium. The monoclonal anti-HER2 antibody, 2C4, was expressed and purified from a mouse hybridoma cell line obtained from ATCC (Manassas, VA). Drugs for in vivo studies were prepared in 10% captisol.

MacNeil I.A., Khan S.A., Sen A., Soltani S.M., Burns D.J., Sullivan B.F, & Laing L.G. (2022). Functional signaling test identifies HER2 negative breast cancer patients who may benefit from c-Met and pan-HER combination therapy. Cell Communication and Signaling : CCS, 20, 4.

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Tepotinib Inhibits Melanoma Cell Growth

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The human malignant melanoma cell line WM451 was purchased from the Cell Bank of Central South University. The cells were incubated in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.) at 37°C in a humidified incubator with 5% CO₂. Subsequently, different concentrations of tepotinib (1, 2, 5, 10, 20 and 50 ng/ml; Selleck Chemicals) were used to treat WM451 cells for 48 h (13 (link)). For tepotinib + hepatocyte growth factor (HGF; R&D Systems, Inc.)-treated group, 50 ng/ml of HGF (a natural agonist of MET) and 10 ng/ml tepotinib were concurrently administered to WM451 cells for 48 h.

Jing G., Yu F, & Xue H. (2022). Tepotinib suppresses proliferation, invasion, migration, and promotes apoptosis of melanoma cells via inhibiting MET and PI3K/AKT signaling pathways. Oncology Letters, 23(6), 170.

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Evaluation of Kinase Inhibitors

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Tepotinib, AZD1208 and dasatinib were purchased from Selleck Chemicals (Houston, TX, U.S.). Savolitinib was purchased from Med Chem Express (Monmouth Junction, NJ, U.S.). PIM447 (LGH447), crizotinib and cabozantinib were purchased from Selleck Chemicals (Houston, TX, U.S.). Drugs were prepared in dimethyl sulfoxide (DMSO) at a concentration of 10–100 mM stock solutions and stored at −20 °C. Further dilutions were made in culture medium to final concentration before use. Primary antibodies and secondary antibodies used are in Table S1.

Attili I., Bonanno L., Karachaliou N., Bracht J.W., Berenguer J., Codony-Servat C., Codony-Servat J., Aldeguer E., Gimenez-Capitan A., Dal Maso A., Fassan M., Chaib I., Molina-Vila M.A., Passaro A., de Marinis F., Pasello G., Guarneri V., Conte P.F, & Rosell R. (2020). SRC and PIM1 as potential co-targets to overcome resistance in MET deregulated non-small cell lung cancer. Translational Lung Cancer Research, 9(5), 1810-1821.

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Inhibition of Oncogenic Pathways

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Doxazosin mesylate, crizotinib, capmatinib, tepotinib, osimertinib and lazertinib were purchased from Selleckchem (Radnor, PA). Amivantamab was obtained from MedchemExpress (Monmouth Junction, NJ). Triton X-100, propidium iodide, PBS tablet, dimethyl sulfoxide (DMSO) and cOmplete™ protease inhibitor cocktail were obtained from Sigma-Aldrich (St. Louis, MO). The immunoblotting and immunostaining antibodies utilized were obtained as follows: c-MET, phospho-c-MET (Y1234/1235), EGFR, phospho-EGFR (Y1068), MEK, phospho-MEK (S217/221), AKT, phospho-AKT (S473), cleaved caspase-3 (Asp175), cleaved caspase-7 (Asp198), cleaved caspase-8 (Asp391), PARP, JAK2, CD44, OCT4 and SOX2 (Cell Signaling Technology, Beverly, MA); phospho-JAK2 (Y1007/1008), STAT3, phospho-STAT3 (Y705) and P-glycoprotein (Abcam, Cambridge, MA); survivin, cyclin D1 and ALDH1A1 (Santa Cruz Biotechnology, CA); GAPDH (Invitrogen, Carlsbad, CA). The secondary antibodies used were HRP-conjugated anti-rabbit and mouse IgG (Bio-Rad Laboratories Inc, CA) and Alexa Fluor-488 or -594 goat anti-mouse and rabbit IgG (Invitrogen).

Kim S., Park J.M., Park S., Jung E., Ko D., Park M., Seo J., Nam K.D., Kang Y.K., Lee K., Farrand L., Kim Y.J., Kim J.Y, & Seo J.H. (2023). Suppression of TNBC metastasis by doxazosin, a novel dual inhibitor of c-MET/EGFR. Journal of Experimental & Clinical Cancer Research : CR, 42, 292.

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Cell Survival Assay for Targeted Drugs

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The effect of tepotinib (SelleckChem, Houston, TX, USA), bortezomib (SelleckChem), and DMF (FUJIFILM Wako) on cell survival was determined using the trypan blue dye exclusion assay as previously described [44 (link)].

Tsubaki M., Seki S., Takeda T., Chihara A., Arai Y., Morii Y., Imano M., Satou T., Shimomura K, & Nishida S. (2020). The HGF/Met/NF-κB Pathway Regulates RANKL Expression in Osteoblasts and Bone Marrow Stromal Cells. International Journal of Molecular Sciences, 21(21), 7905.

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Evaluating c-MET Inhibitor Tepotinib in Gastric Cell Lines

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The SNU620, MKN45, MKN28, KATO III, and AGS gastric cell lines were obtained from the Korean Cell Line Bank (Seoul, Korea) and maintained in RPMI 1640 supplemented with 10% fetal bovine serum. The cells were cultured in 100% humidity and 5% CO₂ at 37 °C. The c-MET inhibitor drug (tepotinib) was purchased from Selleck Chemicals (Houston, TX, USA). The annexin V-APC/propidium iodide (PI) apoptosis detection kit (Thermo Fisher Scientific, Rockford, IL, USA) was used in the present study.

Sohn S.H., Sul H.J., Kim B., Kim B.J., Kim H.S, & Zang D.Y. (2020). Tepotinib Inhibits the Epithelial–Mesenchymal Transition and Tumor Growth of Gastric Cancers by Increasing GSK3β, E-Cadherin, and Mucin 5AC and 6 Levels. International Journal of Molecular Sciences, 21(17), 6027.

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