Synthesis, Screening and Characterization of Novel Potent Arp2/3 Inhibitory Compounds Analogous to CK-666
Branched actin networks polymerized by the Actin-related protein 2 and 3 (Arp2/3) complex play key roles in force generation and membrane remodeling. These networks are particularly important for cell migration, where they drive membrane protrusions of lamellipodia. Several Arp2/3 inhibitory compoun...
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| Format: | Article |
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Frontiers Media S.A.
2022-05-01
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| Series: | Frontiers in Pharmacology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fphar.2022.896994/full |
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| author | Artem I. Fokin Roman N. Chuprov-Netochin Alexander S. Malyshev Alexander S. Malyshev Stéphane Romero Marina N. Semenova Leonid D. Konyushkin Sergey V. Leonov Victor V. Semenov Alexis M. Gautreau Alexis M. Gautreau |
| author_facet | Artem I. Fokin Roman N. Chuprov-Netochin Alexander S. Malyshev Alexander S. Malyshev Stéphane Romero Marina N. Semenova Leonid D. Konyushkin Sergey V. Leonov Victor V. Semenov Alexis M. Gautreau Alexis M. Gautreau |
| author_sort | Artem I. Fokin |
| collection | DOAJ |
| description | Branched actin networks polymerized by the Actin-related protein 2 and 3 (Arp2/3) complex play key roles in force generation and membrane remodeling. These networks are particularly important for cell migration, where they drive membrane protrusions of lamellipodia. Several Arp2/3 inhibitory compounds have been identified. Among them, the most widely used is CK-666 (2-Fluoro-N-[2-(2-methyl-1H-indol-3-yl)ethyl]-benzamide), whose mode of action is to prevent Arp2/3 from reaching its active conformation. Here 74 compounds structurally related to CK-666 were screened using a variety of assays. The primary screen involved EdU (5-ethynyl-2′-deoxyuridine) incorporation in untransformed MCF10A cells. The resulting nine positive hits were all blocking lamellipodial protrusions and cell migration in B16-F1 melanoma cells in secondary screens, showing that cell cycle progression can be a useful read-out of Arp2/3 activity. Selected compounds were also characterized on sea urchin embryos, where Arp2/3 inhibition yields specific phenotypes such as the lack of triradiate spicules and inhibition of archenteron elongation. Several compounds were filtered out due to their toxicity in cell cultures or on sea urchin development. Two CK-666 analogs, 59 (N-{2-[5-(Benzyloxy)-2-methyl-1H-indol-3-yl] ethyl}-3-bromobenzamide) and 69 (2,4-Dichloro-N-[2-(7-chloro-2-methyl-1H-indol-3-yl) ethyl]-5-[(dimethylamino) sulfonyl] benzamide), were active in all assays and significantly more efficient in vivo than CK-666. These best hits with increased in vivo potency were, however, slightly less efficient in vitro than CK-666 in the classical pyrene-actin assay. Induced-fit docking of selected compounds and their possible metabolites revealed interaction with Arp2/3 that suppresses Arp2/3 activation. The data obtained in our screening validated the applicability of original assays for Arp2/3 activity. Several previously unexplored CK-666 structural analogs were found to suppress Arp2/3 activation, and two of them were identified as Arp2/3 inhibitors with improved in vivo efficiency. |
| first_indexed | 2024-04-14T00:25:56Z |
| format | Article |
| id | doaj.art-bcbf0001405b4855b2adee551bb48ce7 |
| institution | Directory Open Access Journal |
| issn | 1663-9812 |
| language | English |
| last_indexed | 2024-04-14T00:25:56Z |
| publishDate | 2022-05-01 |
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| series | Frontiers in Pharmacology |
| spelling | doaj.art-bcbf0001405b4855b2adee551bb48ce72022-12-22T02:22:45ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122022-05-011310.3389/fphar.2022.896994896994Synthesis, Screening and Characterization of Novel Potent Arp2/3 Inhibitory Compounds Analogous to CK-666Artem I. Fokin0Roman N. Chuprov-Netochin1Alexander S. Malyshev2Alexander S. Malyshev3Stéphane Romero4Marina N. Semenova5Leonid D. Konyushkin6Sergey V. Leonov7Victor V. Semenov8Alexis M. Gautreau9Alexis M. Gautreau10CNRS UMR7654, Ecole Polytechnique, Institut Polytechnique de Paris, Palaiseau, FranceDepartment of Molecular and Bio Physics, Moscow Institute of Physics and Technology, Dolgoprudny, RussiaLomonosov Moscow State University, Faculty of Medicine, Moscow, RussiaDukhov Research Institute of Automatics (VNIIA), Moscow, RussiaCNRS UMR7654, Ecole Polytechnique, Institut Polytechnique de Paris, Palaiseau, FranceN. K. Koltzov Institute of Developmental Biology RAS, Moscow, RussiaN. D. Zelinsky Institute of Organic Chemistry RAS, Moscow, RussiaDepartment of Molecular and Bio Physics, Moscow Institute of Physics and Technology, Dolgoprudny, RussiaN. D. Zelinsky Institute of Organic Chemistry RAS, Moscow, RussiaCNRS UMR7654, Ecole Polytechnique, Institut Polytechnique de Paris, Palaiseau, FranceCenter of Molecular and Cellular Biology, Skolkovo Institute of Science and Technology, Moscow, RussiaBranched actin networks polymerized by the Actin-related protein 2 and 3 (Arp2/3) complex play key roles in force generation and membrane remodeling. These networks are particularly important for cell migration, where they drive membrane protrusions of lamellipodia. Several Arp2/3 inhibitory compounds have been identified. Among them, the most widely used is CK-666 (2-Fluoro-N-[2-(2-methyl-1H-indol-3-yl)ethyl]-benzamide), whose mode of action is to prevent Arp2/3 from reaching its active conformation. Here 74 compounds structurally related to CK-666 were screened using a variety of assays. The primary screen involved EdU (5-ethynyl-2′-deoxyuridine) incorporation in untransformed MCF10A cells. The resulting nine positive hits were all blocking lamellipodial protrusions and cell migration in B16-F1 melanoma cells in secondary screens, showing that cell cycle progression can be a useful read-out of Arp2/3 activity. Selected compounds were also characterized on sea urchin embryos, where Arp2/3 inhibition yields specific phenotypes such as the lack of triradiate spicules and inhibition of archenteron elongation. Several compounds were filtered out due to their toxicity in cell cultures or on sea urchin development. Two CK-666 analogs, 59 (N-{2-[5-(Benzyloxy)-2-methyl-1H-indol-3-yl] ethyl}-3-bromobenzamide) and 69 (2,4-Dichloro-N-[2-(7-chloro-2-methyl-1H-indol-3-yl) ethyl]-5-[(dimethylamino) sulfonyl] benzamide), were active in all assays and significantly more efficient in vivo than CK-666. These best hits with increased in vivo potency were, however, slightly less efficient in vitro than CK-666 in the classical pyrene-actin assay. Induced-fit docking of selected compounds and their possible metabolites revealed interaction with Arp2/3 that suppresses Arp2/3 activation. The data obtained in our screening validated the applicability of original assays for Arp2/3 activity. Several previously unexplored CK-666 structural analogs were found to suppress Arp2/3 activation, and two of them were identified as Arp2/3 inhibitors with improved in vivo efficiency.https://www.frontiersin.org/articles/10.3389/fphar.2022.896994/fullArp2/3branched actinArp2/3 inhibitorsCK-666actin polymerization |
| spellingShingle | Artem I. Fokin Roman N. Chuprov-Netochin Alexander S. Malyshev Alexander S. Malyshev Stéphane Romero Marina N. Semenova Leonid D. Konyushkin Sergey V. Leonov Victor V. Semenov Alexis M. Gautreau Alexis M. Gautreau Synthesis, Screening and Characterization of Novel Potent Arp2/3 Inhibitory Compounds Analogous to CK-666 Frontiers in Pharmacology Arp2/3 branched actin Arp2/3 inhibitors CK-666 actin polymerization |
| title | Synthesis, Screening and Characterization of Novel Potent Arp2/3 Inhibitory Compounds Analogous to CK-666 |
| title_full | Synthesis, Screening and Characterization of Novel Potent Arp2/3 Inhibitory Compounds Analogous to CK-666 |
| title_fullStr | Synthesis, Screening and Characterization of Novel Potent Arp2/3 Inhibitory Compounds Analogous to CK-666 |
| title_full_unstemmed | Synthesis, Screening and Characterization of Novel Potent Arp2/3 Inhibitory Compounds Analogous to CK-666 |
| title_short | Synthesis, Screening and Characterization of Novel Potent Arp2/3 Inhibitory Compounds Analogous to CK-666 |
| title_sort | synthesis screening and characterization of novel potent arp2 3 inhibitory compounds analogous to ck 666 |
| topic | Arp2/3 branched actin Arp2/3 inhibitors CK-666 actin polymerization |
| url | https://www.frontiersin.org/articles/10.3389/fphar.2022.896994/full |
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