title: Combination screening in vitro identifies synergistically acting KP372-1 and cytarabine against acute myeloid leukemia.
abstract: Cytogenetic lesions often alter kinase signaling in acute myeloid leukemia (AML) and the addition of kinase inhibitors to the treatment arsenal is of interest. We have screened a kinase inhibitor library and performed combination testing to find promising drug-combinations for synergistic killing of AML cells. Cytotoxicity of 160 compounds in the library InhibitorSelect™ 384-Well Protein Kinase Inhibitor I was measured using the fluorometric microculture cytotoxicity assay (FMCA) in three AML cell lines. The 15 most potent substances were evaluated for dose-response. The 6 most cytotoxic compounds underwent combination synergy analysis based on the FMCA readouts after either simultaneous or sequential drug addition in AML cell lines. The 4 combinations showing the highest level of synergy were evaluated in 5 primary AML samples. Synergistic calculations were performed using the combination interaction analysis package COMBIA, written in R, using the Bliss independence model. Based on obtained results, an iterative combination search was performed using the therapeutic algorithmic combinatorial screen (TACS) algorithm. Of 160 substances, cell survival was ⩽50% at <0.5μM for Cdk/Crk inhibitor, KP372-1, synthetic fascaplysin, herbimycin A, PDGF receptor tyrosine kinase inhibitor IV and reference-drug cytarabine. KP372-1, synthetic fascaplysin or herbimycin A obtained synergy when combined with cytarabine in AML cell lines MV4-11 and HL-60. KP372-1 added 24h before cytarabine gave similar results in patient cells. The iterative search gave further improved synergy between cytarabine and KP372-1. In conclusion, our in vitro studies suggest that combining KP372-1 and cytarabine is a potent and synergistic drug combination in AML.
authors: Department of Medical Sciences, Uppsala University, Box 256, 751 05 Uppsala, Sweden. Electronic address: albin.osterroos@medsci.uu.se.
authors: A
authors: A
authors: Österroos
authors: Department of Medical Sciences, Uppsala University, Box 256, 751 05 Uppsala, Sweden.
authors: M
authors: M
authors: Kashif
authors: Department of Medical Sciences, Uppsala University, Box 256, 751 05 Uppsala, Sweden.
authors: C
authors: C
authors: Haglund
authors: Department of Medical Sciences, Uppsala University, Box 256, 751 05 Uppsala, Sweden.
authors: K
authors: K
authors: Blom
authors: Department of Medical Sciences, Uppsala University, Box 256, 751 05 Uppsala, Sweden.
authors: M
authors: M
authors: Höglund
authors: Department of Medical Sciences, Uppsala University, Box 256, 751 05 Uppsala, Sweden.
authors: C
authors: C
authors: Andersson
authors: Department of Medical Sciences, Uppsala University, Box 256, 751 05 Uppsala, Sweden.
authors: M G
authors: MG
authors: Gustafsson
authors: Department of Medical Sciences, Uppsala University, Box 256, 751 05 Uppsala, Sweden.
authors: A
authors: A
authors: Eriksson
authors: Department of Medical Sciences, Uppsala University, Box 256, 751 05 Uppsala, Sweden.
authors: R
authors: R
authors: Larsson
mesh_terms: D000328:Adult; D000964:Antimetabolites, Antineoplastic / Q000737:chemistry / Q000494:pharmacology; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003561:Cytarabine / Q000819:agonists* / Q000494:pharmacology; D004354:Drug Screening Assays, Antitumor; D004357:Drug Synergism; D005260:Female; D019793:Fluorescein / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D006576:Heterocyclic Compounds, 4 or More Rings / Q000494:pharmacology*; D057166:High-Throughput Screening Assays; D006801:Humans; D020128:Inhibitory Concentration 50; D015470:Leukemia, Myeloid, Acute / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D008297:Male; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors* / Q000378:metabolism; D054852:Small Molecule Libraries; D013050:Spectrometry, Fluorescence; D013777:Tetrazoles / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
chemical_list: D000964:Antimetabolites, Antineoplastic; D005456:Fluorescent Dyes; D006576:Heterocyclic Compounds, 4 or More Rings; C501199:KP372-1; D047428:Protein Kinase Inhibitors; D054852:Small Molecule Libraries; D013777:Tetrazoles; D003561:Cytarabine; D051057:Proto-Oncogene Proteins c-akt; D019793:Fluorescein
keywords: Acute myeloid leukemia; Drug development; KP372-1; Synergy; Tyrosine kinase inhibitor