Lichitsky, Boris V. Komogortsev, Andrey N. Dudinov, Arkady A. Krayushkin, Mikhail M. Khodot, Evgenii N. Samet, Alexander V. Silyanova, Eugenia A. Konyushkin, Leonid D. Karpov, Alexei S. Gorses, Delphine Radimerski, Thomas Semenova, Marina N. Kiselyov, Alex S. Semenov, Victor V. Benzimidazolyl-pyrazolo[3,4‑<i>b</i>]pyridinones, Selective Inhibitors of MOLT‑4 Leukemia Cell Growth and Sea Urchin Embryo Spiculogenesis: Target Quest 1,3-Substituted pyrazolo­[3,4-<i>b</i>]­pyridinones <b>11</b>–<b>18</b> were synthesized by a three-component condensation of Meldrum’s acid with aryl aldehydes and 1,3-substituted 5-aminopyrazoles. Their biological activity was evaluated using the <i>in vivo</i> phenotypic sea urchin embryo assay and the <i>in vitro</i> cytotoxicity screen against human cancer cell lines. In the sea urchin embryo model, 1-benzimidazolyl-pyrazolo­[3,4-<i>b</i>]­pyridinones <b>11</b> caused inhibition of hatching and spiculogenesis at sub-micromolar concentrations. These compounds also selectively and potently inhibited growth of the MOLT-4 leukemia cell line. Subsequent structure–activity relationship studies determined the benzimidazolyl fragment as an essential pharmacophore for both effects. We applied numerous techniques for target identification. A preliminary QSAR target identification search did not result in tangible leads. Attempts to prepare a relevant photoaffinity probe that retained potency in both assays were not successful. Compounds <b>11</b> were further characterized for their activity in a wild-type versus Notch-mutant leukemia cell lines, and in <i>in vitro</i> panels of kinases and matrix metalloproteinases. Using a series of diverse modulators of spiculogenesis as standards, we excluded multiple signaling networks including Notch, Wnt/β-catenin, receptor tyrosine kinases (VEGF/VEGFR, FGF/FGFR), PI3K, and Raf-MEK-ERK as possible targets of <b>11</b>. On the other hand, matrix metalloproteinase-9/hatching enzyme was identified as one potential target. kinase;MOLT -4 leukemia cell line;VEGF;FGF;three-component condensation;photoaffinity probe;vivo phenotypic sea urchin embryo assay;sub-micromolar concentrations;Selective Inhibitors;3K;sea urchin embryo model;target identification;PI;Sea Urchin Embryo Spiculogenesis;cytotoxicity screen;benzimidazolyl fragment;QSAR target identification search;Notch-mutant leukemia cell lines;spiculogenesi;aryl aldehydes;pyridinone;5- aminopyrazoles;cancer cell lines;matrix metalloproteinases;Compounds 11 2019-11-14
    https://acs.figshare.com/articles/journal_contribution/Benzimidazolyl-pyrazolo_3_4_i_b_i_pyridinones_Selective_Inhibitors_of_MOLT_4_Leukemia_Cell_Growth_and_Sea_Urchin_Embryo_Spiculogenesis_Target_Quest/10305338
10.1021/acscombsci.9b00135.s001