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ac220 cat. #5793  (ApexBio)
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Potency comparison of FLT3 inhibitors in cellular assays
Ac220 Cat. #5793, supplied by ApexBio, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Potency comparison of FLT3 inhibitors in cellular assays

Journal: Theranostics

Article Title: LT-171-861, a novel FLT3 inhibitor, shows excellent preclinical efficacy for the treatment of FLT3 mutant acute myeloid leukemia

doi: 10.7150/thno.46593

Figure Lengend Snippet: Potency comparison of FLT3 inhibitors in cellular assays

Article Snippet: Sorafenib (Cat. #3009) and AC220 (Cat. #5793) were purchased from ApexBio Company (Houston, TX).

Techniques: Comparison

Potent pro-apoptotic ability of LT-171-861 in leukemia cells harboring FLT3-ITD mutation. ( A ) MOLM13 cells and MV4-11 cells were respectively incubated with indicated concentrations of LT-171-861, sorafenib and AC220 for 2 h. Cell lysates were used to perform western blot assay. ( B ) MV4-11 cells and MOLM13 cells were treated with LT-171-861 for 36 h. Cells then stained with propidium iodide (PI) /annexin V-FITC and then evaluated by flow cytometry. Annexin V + /PI - and annexin V + /PI + cells were considered dead cells and shown in the bottom chart (n=3). ( C ) MOLM13 cells and MV4-11 cells were treated with indicated concentrations of LT-171-861 for 36 h. PARP, caspase 3, caspase 8, Mcl-1 were evaluated in the cell lysate by western blot. ( D ) MV4-11 cells were treated with LT-171-861 (200 nM), AC220 (200nM) or sorafenib (200 nM) for indicated time periods (0, 4, 8, 12, 24, and 36 h) respectively. Then Cells were stained with propidium iodide/annexin V-FITC and evaluated by flow cytometry. ( E ) MOLM13 cells and MV4-11 cells were treated with cytarabine, LT-171-861 or a combination of cytarabine and LT-171-861 (ratio of 10:1), and cell viability was measured by MTT assay. Error bars show standard deviation (SD), and the combination index (CI) value was calculated by CompuSyn 1.0 software.

Journal: Theranostics

Article Title: LT-171-861, a novel FLT3 inhibitor, shows excellent preclinical efficacy for the treatment of FLT3 mutant acute myeloid leukemia

doi: 10.7150/thno.46593

Figure Lengend Snippet: Potent pro-apoptotic ability of LT-171-861 in leukemia cells harboring FLT3-ITD mutation. ( A ) MOLM13 cells and MV4-11 cells were respectively incubated with indicated concentrations of LT-171-861, sorafenib and AC220 for 2 h. Cell lysates were used to perform western blot assay. ( B ) MV4-11 cells and MOLM13 cells were treated with LT-171-861 for 36 h. Cells then stained with propidium iodide (PI) /annexin V-FITC and then evaluated by flow cytometry. Annexin V + /PI - and annexin V + /PI + cells were considered dead cells and shown in the bottom chart (n=3). ( C ) MOLM13 cells and MV4-11 cells were treated with indicated concentrations of LT-171-861 for 36 h. PARP, caspase 3, caspase 8, Mcl-1 were evaluated in the cell lysate by western blot. ( D ) MV4-11 cells were treated with LT-171-861 (200 nM), AC220 (200nM) or sorafenib (200 nM) for indicated time periods (0, 4, 8, 12, 24, and 36 h) respectively. Then Cells were stained with propidium iodide/annexin V-FITC and evaluated by flow cytometry. ( E ) MOLM13 cells and MV4-11 cells were treated with cytarabine, LT-171-861 or a combination of cytarabine and LT-171-861 (ratio of 10:1), and cell viability was measured by MTT assay. Error bars show standard deviation (SD), and the combination index (CI) value was calculated by CompuSyn 1.0 software.

Article Snippet: Sorafenib (Cat. #3009) and AC220 (Cat. #5793) were purchased from ApexBio Company (Houston, TX).

Techniques: Mutagenesis, Incubation, Western Blot, Staining, Flow Cytometry, MTT Assay, Standard Deviation, Software

Inhibition of FLT3 in BaF3 cell models and in stroma-protecting setting. ( A ) Indicated FLT3 mutations were expressed in BaF3 cells respectively. Transformed BaF3 cells were incubated for 2 h with indicated concentrations of LT-171-861, and ( B-E ) for direct comparison with 3 inhibitors. The autophosphorylation level of FLT3 was visualized and compared by western blot analysis. (E) MOLM13 cells were treated with indicated concentrations of LT-171-861, sorafenib and AC220 for 72 h in condition medium (CM) and normal culture medium (NM). The CM was prepared from HS-5 cell culture for 4 days under routine culture conditions, clarified by centrifugation, and used immediately. The CM was added to complete medium at a final concentration of 40%. Cell viabilities were measured by MTT assay. Data were shown as mean±SD. ( F ) MOLM13 cells were incubated for 8 h in CM with indicated concentrations of LT-171-861 and AC220. The FLT3 phosphorylation levels of FLT3, STAT5 and ERK 1/2 were determined by Western blot.

Journal: Theranostics

Article Title: LT-171-861, a novel FLT3 inhibitor, shows excellent preclinical efficacy for the treatment of FLT3 mutant acute myeloid leukemia

doi: 10.7150/thno.46593

Figure Lengend Snippet: Inhibition of FLT3 in BaF3 cell models and in stroma-protecting setting. ( A ) Indicated FLT3 mutations were expressed in BaF3 cells respectively. Transformed BaF3 cells were incubated for 2 h with indicated concentrations of LT-171-861, and ( B-E ) for direct comparison with 3 inhibitors. The autophosphorylation level of FLT3 was visualized and compared by western blot analysis. (E) MOLM13 cells were treated with indicated concentrations of LT-171-861, sorafenib and AC220 for 72 h in condition medium (CM) and normal culture medium (NM). The CM was prepared from HS-5 cell culture for 4 days under routine culture conditions, clarified by centrifugation, and used immediately. The CM was added to complete medium at a final concentration of 40%. Cell viabilities were measured by MTT assay. Data were shown as mean±SD. ( F ) MOLM13 cells were incubated for 8 h in CM with indicated concentrations of LT-171-861 and AC220. The FLT3 phosphorylation levels of FLT3, STAT5 and ERK 1/2 were determined by Western blot.

Article Snippet: Sorafenib (Cat. #3009) and AC220 (Cat. #5793) were purchased from ApexBio Company (Houston, TX).

Techniques: Inhibition, Transformation Assay, Incubation, Comparison, Western Blot, Cell Culture, Centrifugation, Concentration Assay, MTT Assay, Phospho-proteomics

PK/PD and preliminary toxicity evaluations of LT-171-861. ( A ) Concentrations of LT-171-861 in plasma collected at indicated time points were measured by HPLC/LC-MAS analysis. Blue square and solid line represent intravenous injection administration. ( B ) PD study of LT-171-861 in BALB/c nude mice bearing tumors (120 mm 3 ). Mice were sacrificed at indicated time points and then tumor tissues were dissected and lysed. Phosphorylation levels of FLT3, STAT5 and ERK1/2 were measured by western blotting. ( C ) Bone marrow mononuclear cells (BMMCs) were isolated from healthy C57BL/6 mice. Cells were treated with indicated concentrations of LT-171-861. Apoptosis assays were conducted after 36-hour incubation. ( D ) Healthy C57BL/6 mice (n=6) were intravenously injected with vehicle or LT-171-861 (10 mg/kg) every other day and were orally administered with AC220 (20 mg/kg) or sorafenib (20 mg/kg) once per day for 18 days. 24 h after the last administration, c-Kit + cells were evaluated by flow cytometry. ( E ) PBMCs collected from 3 healthy donors, were incubated with LT-171-861 at indicated concentrations for 72 h and cell viabilities were conducted to evaluate by MTT assay. Data were shown with mean±SD.

Journal: Theranostics

Article Title: LT-171-861, a novel FLT3 inhibitor, shows excellent preclinical efficacy for the treatment of FLT3 mutant acute myeloid leukemia

doi: 10.7150/thno.46593

Figure Lengend Snippet: PK/PD and preliminary toxicity evaluations of LT-171-861. ( A ) Concentrations of LT-171-861 in plasma collected at indicated time points were measured by HPLC/LC-MAS analysis. Blue square and solid line represent intravenous injection administration. ( B ) PD study of LT-171-861 in BALB/c nude mice bearing tumors (120 mm 3 ). Mice were sacrificed at indicated time points and then tumor tissues were dissected and lysed. Phosphorylation levels of FLT3, STAT5 and ERK1/2 were measured by western blotting. ( C ) Bone marrow mononuclear cells (BMMCs) were isolated from healthy C57BL/6 mice. Cells were treated with indicated concentrations of LT-171-861. Apoptosis assays were conducted after 36-hour incubation. ( D ) Healthy C57BL/6 mice (n=6) were intravenously injected with vehicle or LT-171-861 (10 mg/kg) every other day and were orally administered with AC220 (20 mg/kg) or sorafenib (20 mg/kg) once per day for 18 days. 24 h after the last administration, c-Kit + cells were evaluated by flow cytometry. ( E ) PBMCs collected from 3 healthy donors, were incubated with LT-171-861 at indicated concentrations for 72 h and cell viabilities were conducted to evaluate by MTT assay. Data were shown with mean±SD.

Article Snippet: Sorafenib (Cat. #3009) and AC220 (Cat. #5793) were purchased from ApexBio Company (Houston, TX).

Techniques: Clinical Proteomics, Injection, Phospho-proteomics, Western Blot, Isolation, Incubation, Flow Cytometry, MTT Assay

Inhibitory efficacy of LT-171-861 against patient blasts. ( A ) AML blasts harboring FLT3-ITD (patient 1 and 5) and FLT3-wt (patient 2-4 and 6) were treated with indicated concentrations of LT-171-861 for 36 h. Cells then stained with propidium iodide (PI)/annexin V and then evaluated by flow cytometry. ( B ) AML patient blasts expressing FLT3-ITD (patient 7, 9 and 10) were incubated with indicated concentrations of LT-171-861, sorafenib and AC220 for 72 h and cell viabilities were evaluated with MTT assay. ( C ) Blasts from AML patients harboring FLT3-ITD (patient 7 and 8) were incubated with LT-171-861 for 2 h, and p-FLT3, p-STAT5 and p-ERK1/2 were tested by western blot.

Journal: Theranostics

Article Title: LT-171-861, a novel FLT3 inhibitor, shows excellent preclinical efficacy for the treatment of FLT3 mutant acute myeloid leukemia

doi: 10.7150/thno.46593

Figure Lengend Snippet: Inhibitory efficacy of LT-171-861 against patient blasts. ( A ) AML blasts harboring FLT3-ITD (patient 1 and 5) and FLT3-wt (patient 2-4 and 6) were treated with indicated concentrations of LT-171-861 for 36 h. Cells then stained with propidium iodide (PI)/annexin V and then evaluated by flow cytometry. ( B ) AML patient blasts expressing FLT3-ITD (patient 7, 9 and 10) were incubated with indicated concentrations of LT-171-861, sorafenib and AC220 for 72 h and cell viabilities were evaluated with MTT assay. ( C ) Blasts from AML patients harboring FLT3-ITD (patient 7 and 8) were incubated with LT-171-861 for 2 h, and p-FLT3, p-STAT5 and p-ERK1/2 were tested by western blot.

Article Snippet: Sorafenib (Cat. #3009) and AC220 (Cat. #5793) were purchased from ApexBio Company (Houston, TX).

Techniques: Staining, Flow Cytometry, Expressing, Incubation, MTT Assay, Western Blot