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Quizartinib

Manufactured by LC Laboratories
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Quizartinib is a tyrosine kinase inhibitor that targets the FLT3 receptor. It is used for research purposes.

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Lab products found in correlation

Midostaurin, by LC Laboratories (3 mentions) Sorafenib, by LC Laboratories (3 mentions) Lestaurtinib, by LC Laboratories (3 mentions) Recombinant human serum albumin, by Merck Group (2 mentions) Mifepristone, by Merck Group (2 mentions) Sterile pbs, by Thermo Fisher Scientific (2 mentions) Lyophilized bovine plasma, by Merck Group (2 mentions) Trihexyphenidyl hydrochloride, by Selleck Chemicals (2 mentions) Flt3 s 18 antibody, by Santa Cruz Biotechnology (2 mentions) Rpmi 1640, by Thermo Fisher Scientific (2 mentions) Rpmi 1640, by Merck Group (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Entospletinib, by Selleck Chemicals (1 mentions) Venetoclax, by Chemietek (1 mentions) Gilteritinib, by Chemietek (1 mentions) Dasatinib, by LC Laboratories (1 mentions) Arsenic trioxide, by Merck Group (1 mentions) 8 anilinonaphthalene 1 sulfonic acid ans, by Merck Group (1 mentions) Plx51107, by Plexxikon (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions)

7 protocols using quizartinib

1

Inhibitor Compound Preparation Protocol

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ARQ 531 was synthesized and provided by ArQule, Inc., (Burlington, MA). Quizartinib, midostaurin, and dasatinib were purchased from LC Laboratories (Woburn, MA). Gilteritinib and venetoclax were purchased from Chemietek (Indianapolis, IN). Entospletinib (GS-9973) was purchased from Selleckchem (Houston, TX). All chemical compounds were dissolved in Dimethyl sulfoxide (DMSO) to obtain a 10 mM stock solution.
Elgamal O.A., Mehmood A., Jeon J.Y., Carmichael B., Lehman A., Orwick S.J., Truxall J., Goettl V.M., Wasmuth R., Tran M., Mitchell S., Lapalombella R., Eathiraj S., Schwartz B., Stegmaier K., Baker S.D., Hertlein E, & Byrd J.C. (2020). Preclinical efficacy for a novel tyrosine kinase inhibitor, ArQule 531 against acute myeloid leukemia. Journal of Hematology & Oncology, 13, 8.
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2

Preparation and Characterization of FLT3 Inhibitor Compounds

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AGP purified from human or bovine plasma (Sigma) was resuspended in unsupplemented RPMI1640 (Gibco) at 10 mg/mL for working stocks. Recombinant human serum albumin (Sigma) was resuspended in RPMI1640 at 100 mg/mL. Lyophilized bovine plasma (Sigma) was reconstituted in sterile PBS (Gibco). TTT-3002 was a generous gift of TauTaTis, Inc. Lestaurtinib, midostaurin, sorafenib, and quizartinib were purchased from LC Laboratories. Each FLT3 TKI was dissolved at 10 mmol/L in 100% sterile-filtered dimethylsulfoxide (DMSO, Sigma). Mifepristone (Sigma) was dissolved at 100 mmol/L in 100% DMSO. Trihexyphenidyl hydrochloride (Selleckchem) was dissolved at 100 mmol/L in methanol. For cell-based assays, working stocks of 10 μmol/L were prepared for each drug in RPMI1640 supplemented with 0.1% DMSO and 0.2% BSA. For spectrophotometric studies, working stocks of 10 μmol/L were prepared using PBS without sera or albumin. ANS (Sigma) was dissolved in DMSO at 200 mmol/L and then diluted to 400 μmol/L with PBS (0.2% DMSO, final). Western blot analysis was performed using the FLT3 S-18 antibody (Santa Cruz Biotechnology) and for other proteins as indicated (Cell Signaling Technology).
Young D.J., Nguyen B., Li L., Higashimoto T., Levis M.J., Liu J.O, & Small D. (2021). A Method for Overcoming Plasma Protein Inhibition of Tyrosine Kinase Inhibitors. Blood Cancer Discovery, 2(5), 532-547.
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3

AML Cell Line Culture and Compound Preparation

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AML cell lines were cultured in RPMI1640 medium supplemented with 10% fetal calf serum (FCS) and 1% penicillin/streptomycin. Primary samples were acquired upon Johns Hopkins institutional review board (IRB) approval with written informed consent from all patients and healthy volunteers in accordance with the Declaration of Helsinki. Bone marrow (BM) mononuclear cells (MNCs) were isolated by centrifugation and stored in liquid nitrogen until use.
Arsenic trioxide (Sigma) was dissolved in 1M NaOH at 60°C for 20 min and prepared as a 25mM stock solution. Quizartinib (AC220), sorafenib and lestaurtinib (LC Laboratories) were dissolved in dimethylsulfoxide (DMSO) and prepared as 10mM stock solutions in RPMI with 0.1% DMSO. Antibodies used are listed in Supplementary Table 3.
Nagai K., Hou L., Li L., Nguyen B., Seale T., Shirley C., Ma H., Levis M., Ghiaur G., Duffield A, & Small D. (2018). Combination of ATO with FLT3 TKIs eliminates FLT3/ITD+ leukemia cells through reduced expression of FLT3. Oncotarget, 9(68), 32885-32899.
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4

Characterization of FLT3 Inhibitor Interactions

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AGP purified from human or bovine plasma (Sigma) was resuspended in unsupplemented RPMI 1640 (Gibco) at 10 mg/mL for working stocks. Recombinant human serum albumin (Sigma) was resuspended in RPMI 1640 at 100 mg/mL. Lyophilized bovine plasma (Sigma) was reconstituted in sterile PBS (Gibco). TTT-3002 was a generous gift of TauTaTis, Inc. (San Diego, CA). Lestaurtinib, midostaurin, sorafenib and quizartinib were purchased from LC Laboratories. Each FLT3 TKI was dissolved at 10 mM in 100% sterile-filtered dimethylsulfoxide (DMSO, Sigma). Mifepristone (Sigma) was dissolved at 100 mM in 100% DMSO. Trihexyphenidyl hydrochloride (Selleckchem) was dissolved at 100 mM in methanol. For cell-based assays, working stocks of 10 μM were prepared for each drug in RPMI 1640 supplemented with 0.1% DMSO and 0.2% bovine serum albumin. For spectrophotometric studies, working stocks of 10 μM were prepared using PBS without sera or albumin. 8-anilinonaphthalene1-sulfonic acid (ANS, Sigma) was dissolved in DMSO at 200 mM, and then diluted to 400 μM with PBS (0.2% DMSO, final). Western analysis was performed using the FLT3 S-18 antibody (Santa Cruz), and for other proteins as indicated (Cell Signaling Technology).
Young D.J., Nguyen B., Li L., Higashimoto T., Levis M.J., Liu J.O, & Small D. (2021). A Method for Overcoming Plasma Protein Inhibition of Tyrosine Kinase Inhibitors. Blood Cancer Discovery, 2(5), 532-547.
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5

Cytotoxic Agents Quizartinib and PLX51107

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PLX51107 was synthesized at Plexxikon Inc. (Berkeley, CA, USA), a Daiichi Sankyo subsidiary. Quizartinib was obtained from LC Laboratories (Woburn, MA, USA). Stock solutions in dimethyl sulfoxide (DMSO) were stored at -20°C. Dilutions from the stock solutions were prepared in RPMI (Gibco, Waltham, MA, USA) with 10% fetal bovine serum (FBS) (Gemini Bio Products, Sacramento, CA, USA), penicillin/streptomycin, and 2 M L-glutamine (Gibco, Waltham, MA, USA). The final concentration of DMSO in all experiments was ≤0.1%.
Lee L.Y., Hizukuri Y., Severson P., Powell B., Zhang C., Ma Y., Narahara M., Sumi H., Hernandez D., Rajkhowa T., Bollag G, & Levis M. (2021). A novel combination regimen of BET and FLT3 inhibition for FLT3-ITD acute myeloid leukemia. Haematologica, 106(4), 1022-1033.
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6

Cytarabine, Quizartinib, and Flavopiridol Treatment

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Embryos were soaked in egg water containing cytarabine (Pfizer, Milano, Italy), quizartinib (LC Laboratories, Woburn, MA, USA) or flavopiridol (Santa Cruz, Dallas, TX, USA; sc-202157) for drug treatment. Adult fish were intraperitoneally injected with cytarabine (600–2000 mg/kg) and flavopiridol (30–130 mg/kg) once daily for 4 days. The doses for intraperitoneal injection were based on the trials in murine models,31 (link),32 (link) and 4–20-fold higher doses were applied in adult fish.
Liu W., Wu M., Huang Z., Lian J., Chen J., Wang T., Leung A., Liao Y., Zhang Z., Liu Q., Yen K., Lin S., Zon L., Wen Z., Zhang Y, & Zhang W. (2016). c-myb hyperactivity leads to myeloid and lymphoid malignancies in zebrafish. Leukemia, 31(1), 222-233.
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7

Measuring Cellular Viability with ATP Assay

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ATP-based measurements of cellular viability were performed by plating cells in 200 μL of media in 96-well plates. The number of cells and biological replicates seeded varied depending on the cell line and the duration of the experiment. At the indicated times, 40 μL of CellTiter-Glo reagent (Promega) was added to each well, mixed for 5 min, after which the luminescence was measured on the SpectraMax M5 Luminometer (Molecular Devices). For the drug treatment experiments, FRAX-597, Ruxolitinib, and Selumetinib were obtained from Selleckchem and Quizartinib from LC Laboratories.
Wang T., Yu H., Hughes N.W., Liu B., Kendirli A., Klein K., Chen W.W., Lander E.S, & Sabatini D.M. (2017). Gene Essentiality Profiling Reveals Gene Networks and Synthetic Lethal Interactions with Oncogenic Ras. Cell, 168(5), 890-903.e15.
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