Crizotinib (LC Laboratories), osimertinib (LC Laboratories), or the combination were formulated in 0.5% hydroxypropyl methylcellulose (HPMC) and given by oral gavage at 25 mg/kg daily for 5 days per week. An MSK-LX29-SpCTRE tumor from a mouse fed with dox chow for 4 weeks prior to dissociation was incubated with the rAAV at an MOI of 1.6 million genomic copies/cell for 1 hour at 37C. Cells were washed twice with PBS and then engrafted in a 50% Matrigel mixture into a single flank of 20 6-8 week old female NSG mice. All mice were administered dox chow at the time of engraftment and treatment groups were randomized to 4 (vehicle groups) or 5 (C/O groups) mice once tumors reached 100 mm3. Tumors were collected once they reached 1000-1500 mm3 and genomic analysis was performed as described below.
Osimertinib
Manufactured by LC Laboratories
Sourced in United States
Osimertinib is a small-molecule tyrosine kinase inhibitor. It is a laboratory reagent used for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Afatinib, by LC Laboratories (3 mentions)
Erlotinib, by LC Laboratories (3 mentions)
Neratinib, by LC Laboratories (2 mentions)
Mobocertinib, by MedChemExpress (2 mentions)
Crizotinib, by LC Laboratories (1 mentions)
Capmatinib, by Selleck Chemicals (1 mentions)
Lorlatinib, by Selleck Chemicals (1 mentions)
Trametinib, by ChemScene (1 mentions)
Lapatinib, by Selleck Chemicals (1 mentions)
Captisol, by ChemScene (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Dabrafenib, by LC Laboratories (1 mentions)
Erlotinib, by Roche (1 mentions)
Trametinib, by LC Laboratories (1 mentions)
Vemurafenib, by LC Laboratories (1 mentions)
Dimethyl sulfoxide (dmso), by Carl Roth (1 mentions)
Berberine, by Merck Group (1 mentions)
Lapatinib, by LC Laboratories (1 mentions)
Gefitinib, by Selleck Chemicals (1 mentions)
Agarose powder, by Merck Group (1 mentions)
11 protocols using osimertinib
1
Evaluating Crizotinib and Osimertinib in MSK-LX29-SpCTRE Model
2
Synthesis and Characterization of Anti-Cancer Compounds
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Alectinib was synthesized by Chugai Pharmaceutical Co., Ltd. (Tokyo, Japan). Erlotinib was provided by F. Hoffmann-La Roche Ltd. (Basel, Switzerland). Infrigatinib (BGJ398), osimertinib, neratinib, and dabrafenib were obtained from LC Laboratories (Woburn, MA, USA). AZD4547 and trametinib were obtained from ChemScene (Monmouth Junction, NJ, USA). Lorlatinib, capmatinib, and lapatinib were obtained from Selleck Chemicals (Houston, TX, USA). The anti-cancer compound library was obtained from TargetMol (Boston, MA, USA). All agents were dissolved in dimethyl sulfoxide (DMSO: Sigma-Aldrich, St. Louis, MO, USA) for in vitro assays and in a 6% (w/v) solution of Captisol (ChemScene) for in vivo assays. DMSO and Captisol were used as vehicle controls. Recombinant human FGF1, FGF2, FGF7, and FGF12 proteins were obtained from R&D Systems (Minneapolis, MN, USA) and dissolved in phosphate-buffered saline (PBS, Sigma-Aldrich). Recombinant human FGF9 and FGF18 proteins were obtained from Abcam (Cambridge, UK) and dissolved in distilled water. Recombinant human FGF11 protein was obtained from LifeSpan Biosciences (Seattle, WA, USA) and dissolved in Tris buffer (Horizon Discovery, Cambridge, UK).
3
Cell Culture Drug Preparation Protocol
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For cell culture studies, osimertinib (LC Laboratories, USA, Cat. No. 1421373-65-0), trametinib (LC Laboratories, USA, Cat. No. 871700-17-3), and vemurafenib (LC Laboratories, USA, Cat. No. 918504-65-1) were dissolved in dimethyl sulfoxide (DMSO) (Carl Roth, Germany, Cat. No. 4720.4) to a final stock concentration of 10 mM. Cisplatin (pharmacy of University Hospital of Cologne) was diluted to 3.33 mM in 0.9% NaCl.
4
Inhibition of Cell Signaling Pathways
5
Synthesis and Characterization of TAS2940
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TAS2940 fumarate, hereafter referred to as TAS2940, was used for all our experiments. The doses are presented as free base equivalents. TAS2940 was synthesized at Taiho Pharmaceutical Co., Ltd., as per the procedure described in (Figure S1 ). The structure was confirmed using electrospray ionization–mass spectrometry and nuclear magnetic resonance spectroscopy, and its purity was determined to be greater than 99% by HPLC. Lapatinib, afatinib, neratinib, and osimertinib were purchased from LC Laboratories, Selleck Chemicals, Hangzhou APIChem Technology Co., Ltd, and ChemScene LLC, respectively; tucatinib and poziotinib were obtained from Haoyuan ChemExpress Co., Ltd. Each compound was dissolved in DMSO for in vitro assays. For in vivo experiments, TAS2940 was suspended in 0.5% HPMC, while poziotinib and afatinib were suspended in a vehicle solution as described earlier.13 , 25
6
Establishing Mutant EGFR Ba/F3 Cell Line
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Ba/F3 cells were transduced with lentiviral
particles encoding mutant EGFR. After 48 h, cells were placed in blasticidin-containing
media for 14 days, followed by incubation in IL-3-free media for another
14 days to enable selection of mutant EGFR Ba/F3 cells that were IL-3-independent.
These cells were then expanded and used in subsequent assays. A-431
and NCI-H1975 cells in 10% FBS DMEM + 1x Pen-Strep were purchased
from ATCC, PC-9 cells in 10% FBS RPMI + 1x Pen-Strep were purchased
from Millipore Sigma, Osimertinib was purchased from LC Laboratories,
and Gefitinib was purchased from Selleckchem.
particles encoding mutant EGFR. After 48 h, cells were placed in blasticidin-containing
media for 14 days, followed by incubation in IL-3-free media for another
14 days to enable selection of mutant EGFR Ba/F3 cells that were IL-3-independent.
These cells were then expanded and used in subsequent assays. A-431
and NCI-H1975 cells in 10% FBS DMEM + 1x Pen-Strep were purchased
from ATCC, PC-9 cells in 10% FBS RPMI + 1x Pen-Strep were purchased
from Millipore Sigma, Osimertinib was purchased from LC Laboratories,
and Gefitinib was purchased from Selleckchem.
7
NSCLC Cell Line and PDO Characterization
8
Synthesis and Structural Characterization of DS-1205c
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DS-1205b (chemical name: N-[4-(2-amino-5-{4-[(2R)-1,4-dioxan-2-ylmethoxy]-3-methoxyphenyl}pyridin-3-yl)-3-fluorophenyl]-5-methyl-4′-oxo-1′-(tetrahydro-2H-pyran-4-ylmethyl)-1′,4′-dihydro-2,3′-bipyridine-5′-carboxamide 1 4/5 sulfate trihydrate, molecular formula: C41H42FN5O7·1 4/5H2SO4·3H2O) was synthesized by Daiichi Sankyo Co., Ltd. (Tokyo, Japan). The molecular formula was redefined as a non-stoichiometric sulfate hydrate, DS-1205c, C41H42FN5O7·xH2SO4·yH2O (where x is approximately 1 4/5 and y is approximately 3) for clinical trials. Erlotinib and osimertinib were provided by LC Laboratories (Woburn, MA, USA) and Shanghai Sun-shine Chemical Technology Corporation Ltd. (Shanghai, China), respectively. BGB324 was synthesized by Daiichi Sankyo Co., Ltd. (Tokyo, Japan).
9
Evaluating Mobocertinib and Tyrosine Kinase Inhibitors
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CLN-081 was provided by Taiho Pharmaceutical Co., Ltd. Mobocertinib and pimitespib were purchased from MedChemExpress. Poziotinib, osimertinib, afatinib, and erlotinib were purchased from LC Laboratories.
10
Cell Line Culturing and Compound Evaluation Protocol
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All cell lines were obtained from the ATCC (Manassas, VA, USA). The 4T1 (RRID:CVCL_0125) and H1975 (RRID:CVCL_1511) cell lines were cultured in RPMI-1640 medium supplemented with 2 mM L-Glutamine, 1% penicillin/streptomycin (pen/strep) and 10% v/v Fetal Bovine Serum (FBS); A549 (RRID:CVCL_0023) cells were cultured in Ham’s F12 Nutrient Medium supplemented with 2 mM L-Glutamine, 1% pen/strep and 10% FBS; MDA-MB-231 (RRID:CVCL_0062) and HEK293T (RRID:CVCL_0063) cell lines were cultured in DMEM, High Glucose (4500 mg/L D-glucose) supplemented with 2 mM L-Glutamine, 1% pen/strep and 10% FBS and HCT 116 cells were cultured in McCoy’s 5A Medium supplemented with 2 mM L-Glutamine, 1% pen/strep and 10% FBS. All the cell culture media and supplements including FBS were purchased from Biological Industries (Kibbutz Beit-Haemek, Kibbutz, Israel).
Cells were routinely cultured in 75 cm2 tissue culture flask and kept in a humidified atmosphere with 5% CO2 at 37 °C. All cell lines were tested for mycoplasma contamination every 3 months using commercial PCR-based kit (Biological Industries).
FX9847 was provided by Felicitex Therapeutics, Inc. (Natick, MA, USA); Osimertinib (Cat#O-7200) was purchased from LC Laboratories (Woburn, MA, USA); Irinotecan (Cat# I1406), 5-Azacytidine (Cat#A2385) were purchased from Sigma-Aldrich (St. Louis, MO, USA).
Cells were routinely cultured in 75 cm2 tissue culture flask and kept in a humidified atmosphere with 5% CO2 at 37 °C. All cell lines were tested for mycoplasma contamination every 3 months using commercial PCR-based kit (Biological Industries).
FX9847 was provided by Felicitex Therapeutics, Inc. (Natick, MA, USA); Osimertinib (Cat#O-7200) was purchased from LC Laboratories (Woburn, MA, USA); Irinotecan (Cat# I1406), 5-Azacytidine (Cat#A2385) were purchased from Sigma-Aldrich (St. Louis, MO, USA).
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