Adavosertib, dabrafenib, trametinib, sorafenib and lenvatinib were purchased from Selleck Chemicals. Adavosertib, dabrafenib, trametinib, sorafenib and lenvatinib were each dissolved in dimethyl sulfoxide (DMSO; Sigma) to a concentration of 10 mmol/L for in vitro experiments. For xenograft experiments, Adavosertib and lenvatinib were diluted in methyl cellulose (Sigma) and distilled water (1:200 w/v) to a concentration of 12 mg/ml and stored at −80°C until use. dabrafenib and trametinib were dissolved in 0.5% (hydroxypropyl)methyl cellulose (Sigma), 0.2% Tween 80 (Sigma), and distilled water to concentrations of 8 mg/ml and 0.16 mg/ml, respectively, and stored at −80°C until use. sorafenib was dissolved in 50% Kolliphor EL (Sigma) and 50% ethanol (Sigma) to a concentration of 57.6 mg/mL and stored at −80°C; it was further dissolved at a final concentration of 14.4 mg/mL in water before in vivo use.
Lenvatinib
Manufactured by Merck Group
Sourced in United States
Lenvatinib is a laboratory product manufactured by Merck Group. It is a tyrosine kinase inhibitor used for research and development purposes. Its core function is to inhibit the activity of multiple receptor tyrosine kinases involved in angiogenesis and tumor growth.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Tween 80, by Merck Group (2 mentions)
Methylcellulose, by Merck Group (2 mentions)
Sorafenib, by Merck Group (2 mentions)
Kolliphor el, by Merck Group (2 mentions)
Lenvatinib, by Selleck Chemicals (2 mentions)
Trametinib, by Selleck Chemicals (2 mentions)
Hydroxypropyl methylcellulose, by Merck Group (2 mentions)
Trametinib, by Merck Group (2 mentions)
Dabrafenib, by Merck Group (2 mentions)
Adavosertib, by Selleck Chemicals (2 mentions)
Sorafenib, by Selleck Chemicals (2 mentions)
Dabrafenib, by Selleck Chemicals (2 mentions)
Ethanol, by Merck Group (1 mentions)
Adavosertib, by Merck Group (1 mentions)
Epirubicin, by Pfizer (1 mentions)
Iodized oil, by Guerbet (1 mentions)
Annexin 5 fitc kit, by BD (1 mentions)
Facscan flow cytometer, by BD (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
9 protocols using lenvatinib
1
Preparation of Anticancer Drug Formulations
2
Combination TACE and Tyrosine Kinase Inhibitor Therapy
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The treatment protocol was described in a previous study (9 (link)). Each patient received the TACE treatment. Briefly, after local anesthesia with 3–5 ml of 1% lidocaine administered in the groin, hepatic arteriography was performed. Then, 2–20 ml of an injectable suspension of iodized oil (10-20 ml, Guerbet) and epirubicin (40-45 mg, Pfizer) was injected into the target artery using a 2-F microcatheter (Terumo), followed by gelatin sponge particle (350–560 μm, Alicon) embolization until arterial flow stasis was achieved. Treatment was continued until progression to the TACE refractory criteria, unacceptable toxicity, or withdrawal of consent.
Sorafenib (Pfizer) or lenvatinib (Merck) was administered within 7 days before or after TACE. Drug dosage was based on the recommended doses in the SHARP (10 (link)) and REFLECT (7 (link)) studies, respectively: sorafenib: 400 mg orally, twice per day; lenvatinib: 8 mg orally, once per day (body weight < 60 kg) or 12 mg orally, once per day (body weight ≥ 60 kg). Doses were adjusted according to the grade of the National Cancer Institute’s Common Terminology Criteria for Adverse Events (CTCAE version 4.0). lenvatinib or sorafenib was continually administered until disease progression developed or unacceptable adverse events (AEs) appeared.
Sorafenib (Pfizer) or lenvatinib (Merck) was administered within 7 days before or after TACE. Drug dosage was based on the recommended doses in the SHARP (10 (link)) and REFLECT (7 (link)) studies, respectively: sorafenib: 400 mg orally, twice per day; lenvatinib: 8 mg orally, once per day (body weight < 60 kg) or 12 mg orally, once per day (body weight ≥ 60 kg). Doses were adjusted according to the grade of the National Cancer Institute’s Common Terminology Criteria for Adverse Events (CTCAE version 4.0). lenvatinib or sorafenib was continually administered until disease progression developed or unacceptable adverse events (AEs) appeared.
3
Preparation and Formulation of Anticancer Drugs
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Adavosertib, sorafenib, lenvatinib, dabrafenib, and trametinib were purchased from Selleck Chemicals, and diluted in dimethyl sulfoxide (DMSO; Merck) to a concentration of 10 mmol/L and stored at −80 °C until in vitro use. For the in vivo experiments, Adavosertib and lenvatinib were diluted in methyl cellulose (Merck) and distilled water (1:200 w/v) to a final concentration of 12 mg/mL. sorafenib was dissolved in 50/50% Kolliphor EL (Merck) and ethanol (Merck) and further diluted with water to a final concentration of 14.4 mg/mL before use. dabrafenib and trametinib were dissolved in 0.5% (hydroxypropyl) methylcellulose (Merck), 0.2% Tween 80 (Merck), and distilled water to concentrations of 8 mg/mL and 0.16 mg/mL, respectively. All drugs were stored at −80 °C until their in vivo use.
4
Apoptosis Induction in NSCLC Cells
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NSCLC cells (A549) were grown at 37°C until 90% confluences were reached. Cells (1×108) were then incubated with Lenvatinib (2 mg/ml; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) and/or rAd-p53 (1011 pfu) for 48 h at 37°C. After incubation, the tumor cells were trypsinized and collected. The cells were then washed in cold PBS, adjusted to 1×106 cells/ml with PBS, labeled with Annexin V-FITC and PI (Annexin V-FITC kit; BD Biosciences, Franklin Lakes, NJ, USA), and analyzed with a FACScan flow cytometer (BD Biosciences). The treatments were performed in triplicate, and apoptosis was analyzed with a FACScan flow cytometer (BD Biosciences). using CellQuest Pro solfware (v.5.1, BD Biosciences).
5
Hepatocellular Carcinoma Cell Cultivation
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HepG2 and Hep3B HCC cell lines were obtained from the American Type Culture Collection (ATCC; Manassas, VA). HepG2 and Hep3B cells were cultivated in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (Thermo Fisher Scientific, Waltham, MA). Cell lines were regularly tested for mycoplasma infections using Mycoplasma Detection Kit (In vivo Gen, San Diego, CA) as well as regularly changed with thawed stocking cell lines. l -AA, sodium ascorbate, 2′,7-dichlorofluorescein diacetate (DCFH-DA), lenvatinib, propidium iodide (PI), regorafenib, and thiazolyl blue tetrazolium bromide (MTT) were purchased from Sigma Aldrich (St. Louis, MO).
6
Lenvatinib and I-131 Cytotoxicity Assay
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HK-1 cells were incubated with Lenvatinib (5 mg/ml; Sigma-Aldrich; Merck KGaA) or/and I-131 (3 mg/ml; Zhengzhou Double-radioisotope Application Technology Co., Ltd., Zhengzhou, China) in 96-well plates for 48 h in triplicate for each condition. Following incubation, 20 µl MTT (5 mg/ml) in PBS solution was added to each well and was further incubated for 4 h. the majority of the medium was removed and 100 µl dimethyl sulfoxide was added to each wells to dissolve the crystals. The optical density was measured at a wavelength of 450 nm.
7
Examination of Human Liver Cancer Cell Lines
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Five human HCC cell lines (Huh-7, SNU449, PLC-PRF-5, SNU398, and Sk-Hep-1) were examined in this study. SNU449 was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). Huh-7 and PLC-PRF-5 (PLC) were obtained from the RIKEN BioResource Center Cell Bank (Tsukuba, Japan). Dr. Ke, Department of Diabetes Complications and Metabolism, Beckman Research Institute of City of Hope, kindly provided us with SNU398 and Sk-Hep-1 cell lines. All cell lines were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (Gibco, Waltham, MA, USA) and 1% penicillin-streptomycin. All cells were maintained in a 37 °C incubator with a 5% humidified CO2 atmosphere. Curcumin was generously provided by EuroPharma USA, Green Bay, WI, USA, and Lenvatinib was purchased from Sigma-Aldrich, St. Louis, MO, USA.
8
Quantitative Analysis of Lenvatinib and Biochanin A
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Lenvatinib and cyclosporin A were purchased from Sigma-Aldrich (St. Louis, MO, USA). As an internal standard, biochanin A was purchased from Toronto Research Chemicals Inc. (North York, ON, Canada). Polyethylene glycol 400 (PEG 400) and heparin sodium were purchased from Sigma-Aldrich. Pentobarbital sodium was obtained from SCI Pharmtech (Taoyuan, Taiwan). The solvents and reagents for chromatography were purchased from J.T. Baker (Phillipsburg, NJ, USA) and Merck (Darmstadt, Germany). Standard solutions of Lenvatinib and biochanin A in methanol were stored at −20 °C. Triply deionized water from Millipore (Bedford, MA, USA) was used for all preparations.
9
Antioxidant and Anti-Cancer Compound Evaluation
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Melatonin, indole-3-propionic acid and lenvatinib were purchased from Sigma (St. Louis, MO, USA). Sorafenib was purchased from LC Laboratories (Woburn, MA, USA). The LPO-586 kit for LPO was obtained from Enzo Life Science (Farmingdale, NY, USA). All the used chemicals were of analytical grade and came from commercial sources.
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