Selumetinib, carboplatin, irinotecan, and temsirolimus were purchased from MedChem Express. JQ1 was kindly provided by Dr. James Bradner (Dana-Farber Cancer Institute, Boston, MA, USA). Drugs were resuspended in N-Methyl-2-pyrrolidone (NMP) to create a stock solution (Sigma Aldrich) and diluted in PTD buffer (30 % PEG-400; 5 % Tween 80; 65 % Dextrose water, D5W, Sigma Aldrich) before drug dosing.
Selumetinib
Manufactured by MedChemExpress
Sourced in United States
Selumetinib is a small molecule inhibitor that targets the MEK1/2 enzyme. It is a white to off-white solid that is used for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Trametinib, by MedChemExpress (3 mentions)
Cobimetinib, by MedChemExpress (2 mentions)
Erlotinib, by MedChemExpress (2 mentions)
Temsirolimus, by MedChemExpress (1 mentions)
Carboplatin, by MedChemExpress (1 mentions)
Irinotecan, by MedChemExpress (1 mentions)
Alamarblue assay, by Thermo Fisher Scientific (1 mentions)
Ci 1040, by MedChemExpress (1 mentions)
Abt 263, by Merck Group (1 mentions)
Mkc3946, by MedChemExpress (1 mentions)
Mek162, by MedChemExpress (1 mentions)
Buparlisib, by MedChemExpress (1 mentions)
Prism 9, by GraphPad (1 mentions)
Copanlisib, by MedChemExpress (1 mentions)
Tamoxifen, by MedChemExpress (1 mentions)
Human recombinant epidermal growth factor, by Thermo Fisher Scientific (1 mentions)
Cerulein, by MedChemExpress (1 mentions)
Kribb11, by MedChemExpress (1 mentions)
Transforming growth factor alpha, by Thermo Fisher Scientific (1 mentions)
Torkinib, by MedChemExpress (1 mentions)
16 protocols using selumetinib
1
Drug Formulation and Dosing Protocol
2
Anticancer Activity Screening Assay
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AMPC was synthesized in the Department of Studies in Organic Chemistry, University of Mysore20 . Selumetinib (AZD6244), Pimasertib (AS703026), CI-1040 (PD184352), and Trametinib (GSK1120212) were purchased from MedChemExpress (Monmouth Junction, NJ) and re-constituted in DMSO. Cells were seeded at 2000 cells per well in 96-well plates with 2% FBS media and treated with drug for 48 to 72 h. The viability of cells was determined using AlamarBlue assay (ThermoFisher Scientific, Waltham, MA).
3
Multi-Compound Cytotoxicity Screening
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The following chemicals were used: 2c [9 (link)], dimethyl sulfoxide (DMSO), Doxorubicin, MK2206, Torin1, Bafilomycin A1, Chloroquine, ABT263 (Sigma), Gemcitabine, XMD8-92, YKL-06-061, ABT199 (CSN Scientific), Selumetinib, TMP195, MKC3946 (MedChemExpress), SAHA (Cayman Chemicals), NKL54 [46 (link)].
4
Cytotoxicity of MEK Inhibitors
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The in vitro cytotoxicity of MEK162, Selumetinib and Trametinib (MedChem Express, Stockholm, Sweden) was tested by MTS and MTT assays. All inhibitors were weighed, dissolved in dimethyl sulfoxide (DMSO) and stored at -20°C until use. Cytotoxicity assay dilutions were prepared in cell culture medium, keeping final DMSO concentration <0.5%. Final concentrations of the small molecule inhibitors ranged from 50 μM to 0.15 nM, indicated in the respective figures. The in vitro sensitivity of cell lines was assessed by using 4-day MTS conversion assays, as described previously [28 (link)]. In vitro cytotoxicity of patient cells was assessed by using a 4-day MTT conversion assay, as described before [8 (link)]. Data were normalized to vehicle (DMSO) controls.
5
Pharmacokinetics of PI3Ki and MEKi
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The PI3K inhibitor (PI3Ki) buparlisib (BKM120) and the MEK inhibitor (MEKi) selumetinib (AZD6244) were purchased from MedChem Express (Monmouth Junction, NJ) or Chemietek (Indianapolis, IN) and dissolved in dimethyl sulfoxide (DMSO). In single-dose pharmacokinetics studies in human patients, maximum observed plasma concentrations for buparlisib and selumetinib were 2–5 μM (1–2 μg/mL) and 1.1–2.0 μM (0.5–0.9 μg/mL), respectively [39 (link), 40 (link)]. Depending on experimental requirements, drugs were used at or above these clinically relevant dose ranges.
6
Cytotoxicity Profiling of Antitumor Drugs
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Four antitumor drugs including cobimetinib, copanlisib, selumetinib, and tamoxifen were purchased from MedChemExpress (Shanghai, China). For the CCK-8 assay, OS cells in the logarithmic growth phase were plated in 96-well plates and treated with different concentrations of drugs. After 72 h of drug induction, 10 µL CCK-8 solution was added to the cells and incubated for 2.5 h. The optical density (OD) at 490 nm was measured on a microplate reader. The IC50 value was calculated on the GraphPad Prism 9 software by non-linear regression analysis.
7
Cerulein-Induced Pancreatic Injury Model
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Cerulein (40 μg/kg or 5 μg/mouse), KRIBB11 (mainly 2 μM for cells and 50 mg/kg for mice), erlotinib (2 μM for cells and 100 mg/kg for mice), selumetinib (100 nM for cells) and torkinib (50 nM for cells) were purchased from MedChem Express (Monmouth Junction, NJ, USA). Recombinant human epidermal growth factor (EGF mainly 20 ng/ml for cells) and transforming growth factor alpha (mainly TGFα 50 ng/ml for cells) were purchased from PeproTech (Rocky Hill, NJ, USA). All reagents were stored according to the manufacturer’s instructions.
8
Combinatorial Anticancer Drug Screening
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A549 cells were seeded in 96-well plates at a density of 2x103/well and maintained in an incubator overnight. After the removal of the culturing medium, and the cells were fed with fresh medium containing dasatinib, selumetinib, and MK-2206 (MedChemExpress LLC. Monmouth Junction, USA) alone, or in combination. Cell viability was measured by the Dojindo cell counting kit-8 (CCK-8, GlpBio, USA) at 24, 48, and 72 hours.
9
Inhibition of MEK and EGFR Pathways
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E7080 (lenvatinib) was purchased from AdooQ BioScience (Irvine, CA, USA). Selective MEK inhibitors, U0126 for the in vitro study and AZD6244 (selumetinib) for the in vivo study, were purchased from MedChemExpress (Monmouth Junction, NJ, USA) and Selleck Chemicals (Houston, TX, USA), respectively.
10
Organoid-derived Tumor Cell Viability Assay
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KraslslG12D/wt; p53lox/lox and KrasG12D/wt; p53∆/∆ organoids were seeded in 100‐µL 10% Matrigel per 96‐well. Two, 24, and 48 hours after seeding, the organoids were lysed and CellTiter‐Glo 3D Luminescent Cell Viability Assay (Promega, Madison, WI) was performed according to the manufacturer’s protocol.
For inhibitor treatment, KrasG12D/wt; p53∆/∆; LMP_shRenilla.713 tumor cell lines established as 2D or 3D cultures from primary, organoid‐derived tumors were plated at 1,000 cells (2D cell line) and 10,000 cells (3D cell line) per 96‐well and treated with selumetinib (3.36 µM; MedChem Express, Monmouth, NJ), NVP‐BKM120 (0.25 µM; MedChem Express), or a combination of both inhibitors for 24 hours and 48 hours. At the indicated time points, cells were lysed using CellTiter‐Glo Luminescent Cell Viability Assay or CellTiter‐Glo 3D Luminescent Cell Viability Assay, and luminescence was acquired on a Glomax Multi Detection System (Promega, Madison, WI).
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KraslslG12D/wt; p53lox/lox and KrasG12D/wt; p53∆/∆ organoids were seeded in 100‐µL 10% Matrigel per 96‐well. Two, 24, and 48 hours after seeding, the organoids were lysed and CellTiter‐Glo 3D Luminescent Cell Viability Assay (Promega, Madison, WI) was performed according to the manufacturer’s protocol.
For inhibitor treatment, KrasG12D/wt; p53∆/∆; LMP_shRenilla.713 tumor cell lines established as 2D or 3D cultures from primary, organoid‐derived tumors were plated at 1,000 cells (2D cell line) and 10,000 cells (3D cell line) per 96‐well and treated with selumetinib (3.36 µM; MedChem Express, Monmouth, NJ), NVP‐BKM120 (0.25 µM; MedChem Express), or a combination of both inhibitors for 24 hours and 48 hours. At the indicated time points, cells were lysed using CellTiter‐Glo Luminescent Cell Viability Assay or CellTiter‐Glo 3D Luminescent Cell Viability Assay, and luminescence was acquired on a Glomax Multi Detection System (Promega, Madison, WI).
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