The small molecule inhibitor NAV-2729 has a complex target profile including multiple ADP-ribosylation factor regulatory proteins

Author:

Rosenberg, Eric M

Jian, Xiaoying

Soubias,Olivier

Yoon, Hye-Young

Yadav, Mukesh P

Hammoudeh, Sarah

Pallikkuth, Sandeep

Akpan, Itoro

Chen, Pei-Wen

Maity, Tapan K

Jenkins, Lisa M

Yohe,Marielle

Byrd,Robert

Randazzo, Paul A
Author Address

Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute; Bethesda, MD, USA., Center for Structural Biology Laboratory, Center for Cancer Research, National Cancer Institute; Frederick, MD, USA., Department of Biology, Williams College; Williamstown, MA, USA., Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute; Bethesda, MD, USA., Pediatric Oncology Branch, Center for Cancer Research, National Cancer Institute; Bethesda, MD, USA; Laboratory of Cell and Developmental Signaling, Center for Cancer Research; Frederick, MD, USA., Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute; Bethesda, MD, USA. Electronic address: randazzp@mail.nih.gov.,

Abstract:

The ADP-ribosylation factor (Arf)2 GTPases and their regulatory proteins are implicated in cancer progression. NAV-2729 was previously identified as a specific inhibitor of Arf6 that reduced progression of uveal melanoma in an orthotopic xenograft. Here, our goal was to assess the inhibitory effects of NAV-2729 on the proliferation of additional cell types. We found NAV-2729 inhibited proliferation of multiple cell lines, but Arf6 expression did not correlate with NAV-2729 sensitivity, and knockdown of Arf6 affected neither cell viability nor sensitivity to NAV-2729. Furthermore, binding to native Arf6 was not detected; however, we determined that NAV-2729 inhibited both Arf exchange factors and Arf GTPase activating proteins (GAPs). ASAP1, a GAP linked to cancer progression, was further investigated. We demonstrated that NAV-2729 bound to the PH domain of ASAP1 and changed ASAP1 cellular distribution. However, ASAP1 knockdown did not fully recapitulate the cytoskeletal effects of NAV-2729 nor affect cell proliferation. Finally, our screens identified 48 other possible targets of NAV-2729. These results illustrate the complexities of defining targets of small molecules and identify NAV-2729 as a model PH domain- binding inhibitor. Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.

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