Abstract:Members of the Wee family of kinases negatively regulate the cell cycle via phosphorylation of CDK1 and are considered potential drug targets. Herein, we investigated the structure–function relationship of human Wee1, Wee2, and Myt1 (PKMYT1). Purified recombinant full-length proteins and kinase domain constructs differed substantially in phosphorylation states and catalytic competency, suggesting complex mechanisms of activation. A series of crystal structures reveal unique features that distinguish Wee1 and W… Show more
“…At the time of writing this manuscript, the 1.9Å co-crystal structure of the human WEE1 kinase domain in complex with AZD1775 was published (PDB: 5V5Y). [13] The proposed binding orientation of AZD1775 in the ATP-binding site of WEE1 in our computational model was in good agreement with the co-crystal structure (Supporting Information, Figure S4), with only minor differences observed in the orientation of the side chains. Importantly, both our computational model and the WEE1-AZD1775 co-crystal structure equally support the proposed structural modifications to AZD1775 described above.…”
Section: Resultssupporting
confidence: 80%
“…The WEE1-AZD1775 co-crystal structure indicates that the allyl group may have a hydrophobic contact with Lys328. [11c] Further compounds may need to be developed to more thoroughly investigate the scope of replacing the allyl group to include substituents that maintain the electronic distribution of negative electrostatic and hydrophobic fields and that can be accommodated in the small hydrophobic pocket in the ATP-binding site of WEE1.…”
Section: Discussionmentioning
confidence: 99%
“…A kinome interaction network study identified ABL1, LCK, LRRK2, TNK2, and SYK as targets of AZD1775 with Ki values below 1 μM. [11a] Recently, it has been reported that AZD1775 has potent nanomolar activity against PLK1, and that AZD1775 should now be considered a dual WEE1 and PLK1 inhibitor. [11b, 11c] A full kinome profile for AZD1775 across a panel of 468 human kinases at a concentration of 500 n m identified PLK1, FLT3 (D835V), JAK3, GCN2 (S808G), ABL1 (H396P), and JAK2 as additional kinase targets.…”
Section: Discussionmentioning
confidence: 99%
“…[11a] Recently, it has been reported that AZD1775 has potent nanomolar activity against PLK1, and that AZD1775 should now be considered a dual WEE1 and PLK1 inhibitor. [11b, 11c] A full kinome profile for AZD1775 across a panel of 468 human kinases at a concentration of 500 n m identified PLK1, FLT3 (D835V), JAK3, GCN2 (S808G), ABL1 (H396P), and JAK2 as additional kinase targets. [11c] Furthermore, the authors suggested that WEE1 and PLK1 were the most relevant targets for mediating the anticancer effects of AZD1775; however, our studies support that WEE1 inhibition can be achieved with little impact on cytotoxicity.…”
Section: Discussionmentioning
confidence: 99%
“…In preclinical studies, AZD1775 demonstrated potent cytotoxicity in a broad panel of cancer cell lines and led to xenograft tumor growth inhibition; [10] however, these anticancer activities cannot be attributed to WEE1 inhibition alone since AZD1775 is known to inhibit other kinases and may have off-target effects outside of the kinome. [6, 11] The combination of WEE1 inhibitors with DNA-damaging agents has been proposed to be more effective in p53-mutant cancer cells that rely on DNA repair at the G2-M checkpoint. However, several preclinical studies have demonstrated that AZD1775 has anticancer activity independent of p53 function, and this has also been attributed to the alternative functions of WEE1 and/or the off-target effects of AZD1775.…”
WEE1 kinase regulates the G /M cell-cycle checkpoint, a critical mechanism for DNA repair in cancer cells that can confer resistance to DNA-damaging agents. We previously reported a series of pyrazolopyrimidinones based on AZD1775, a known WEE1 inhibitor, as an initial investigation into the structural requirements for WEE1 inhibition. Our lead inhibitor demonstrated WEE1 inhibition in the same nanomolar range as AZD1775, and potentiated the effects of cisplatin in medulloblastoma cells, but had reduced single-agent cytotoxicity. These results prompted the development of a more comprehensive series of WEE1 inhibitors. Herein we report a series of pyrazolopyrimidinones and identify a more potent WEE1 inhibitor than AZD1775 and additional compounds that demonstrate that WEE1 inhibition can be achieved with reduced single-agent cytotoxicity. These studies support that WEE1 inhibition can be uncoupled from the potent cytotoxic effects observed with AZD1775, and this may have important ramifications in the clinical setting where WEE1 inhibitors are used as chemosensitizers for DNA-targeted chemotherapy.
“…At the time of writing this manuscript, the 1.9Å co-crystal structure of the human WEE1 kinase domain in complex with AZD1775 was published (PDB: 5V5Y). [13] The proposed binding orientation of AZD1775 in the ATP-binding site of WEE1 in our computational model was in good agreement with the co-crystal structure (Supporting Information, Figure S4), with only minor differences observed in the orientation of the side chains. Importantly, both our computational model and the WEE1-AZD1775 co-crystal structure equally support the proposed structural modifications to AZD1775 described above.…”
Section: Resultssupporting
confidence: 80%
“…The WEE1-AZD1775 co-crystal structure indicates that the allyl group may have a hydrophobic contact with Lys328. [11c] Further compounds may need to be developed to more thoroughly investigate the scope of replacing the allyl group to include substituents that maintain the electronic distribution of negative electrostatic and hydrophobic fields and that can be accommodated in the small hydrophobic pocket in the ATP-binding site of WEE1.…”
Section: Discussionmentioning
confidence: 99%
“…A kinome interaction network study identified ABL1, LCK, LRRK2, TNK2, and SYK as targets of AZD1775 with Ki values below 1 μM. [11a] Recently, it has been reported that AZD1775 has potent nanomolar activity against PLK1, and that AZD1775 should now be considered a dual WEE1 and PLK1 inhibitor. [11b, 11c] A full kinome profile for AZD1775 across a panel of 468 human kinases at a concentration of 500 n m identified PLK1, FLT3 (D835V), JAK3, GCN2 (S808G), ABL1 (H396P), and JAK2 as additional kinase targets.…”
Section: Discussionmentioning
confidence: 99%
“…[11a] Recently, it has been reported that AZD1775 has potent nanomolar activity against PLK1, and that AZD1775 should now be considered a dual WEE1 and PLK1 inhibitor. [11b, 11c] A full kinome profile for AZD1775 across a panel of 468 human kinases at a concentration of 500 n m identified PLK1, FLT3 (D835V), JAK3, GCN2 (S808G), ABL1 (H396P), and JAK2 as additional kinase targets. [11c] Furthermore, the authors suggested that WEE1 and PLK1 were the most relevant targets for mediating the anticancer effects of AZD1775; however, our studies support that WEE1 inhibition can be achieved with little impact on cytotoxicity.…”
Section: Discussionmentioning
confidence: 99%
“…In preclinical studies, AZD1775 demonstrated potent cytotoxicity in a broad panel of cancer cell lines and led to xenograft tumor growth inhibition; [10] however, these anticancer activities cannot be attributed to WEE1 inhibition alone since AZD1775 is known to inhibit other kinases and may have off-target effects outside of the kinome. [6, 11] The combination of WEE1 inhibitors with DNA-damaging agents has been proposed to be more effective in p53-mutant cancer cells that rely on DNA repair at the G2-M checkpoint. However, several preclinical studies have demonstrated that AZD1775 has anticancer activity independent of p53 function, and this has also been attributed to the alternative functions of WEE1 and/or the off-target effects of AZD1775.…”
WEE1 kinase regulates the G /M cell-cycle checkpoint, a critical mechanism for DNA repair in cancer cells that can confer resistance to DNA-damaging agents. We previously reported a series of pyrazolopyrimidinones based on AZD1775, a known WEE1 inhibitor, as an initial investigation into the structural requirements for WEE1 inhibition. Our lead inhibitor demonstrated WEE1 inhibition in the same nanomolar range as AZD1775, and potentiated the effects of cisplatin in medulloblastoma cells, but had reduced single-agent cytotoxicity. These results prompted the development of a more comprehensive series of WEE1 inhibitors. Herein we report a series of pyrazolopyrimidinones and identify a more potent WEE1 inhibitor than AZD1775 and additional compounds that demonstrate that WEE1 inhibition can be achieved with reduced single-agent cytotoxicity. These studies support that WEE1 inhibition can be uncoupled from the potent cytotoxic effects observed with AZD1775, and this may have important ramifications in the clinical setting where WEE1 inhibitors are used as chemosensitizers for DNA-targeted chemotherapy.
The 4-anilinoquinoline and 4-anilinoquinazoline ring systems have been the focus of significant efforts in prior kinase drug discovery programs, which have led to approved medicines. Broad kinome profiles of these compounds have now been assessed with the advent of advanced screening technologies. These ring systems, while originally designed for specific targets including epidermal growth factor receptor (EGFR), actually display a number of potent collateral kinase targets, some of which have been associated with negative clinical outcomes. We have designed and synthesized a series of 4-anilino-quin(az)olines in order to better understand the structure activity relationships of three main collateral kinase targets of quin(az)oline-based kinase inhibitors: cyclin G associated kinase (GAK), STE20-like serine/threonine-protein kinase (SLK) and Serine/threonine-protein kinase 10 (STK10). This was achieved through a series of quantitative structure activity relationship (QSAR) analysis and water mapping of the kinase ATP binding sites.
Protein kinase, membrane‐associated tyrosine/threonine 1 (PKMYT1), which is associated with progression of tumor, is upregulated in a variety of cancers. However, its expression and the underlying molecular mechanisms in the context of bladder cancer (BLCA) remain elusive. Here we found that PKMYT1 expression was markedly higher expression in BLCA, which was correlated with poorer prognosis compared with low expression. Knockdown of PKMYT1 significantly inhibited the BLCA cells proliferation in vivo and in vitro, and migration and invasion, reduced G2/M phase in cell cycle and induced apoptosis. Mechanically, YAP and TEAD1 knockdown suppressed PKMYT1 expression in BLCA cells, whereas overexpression of YAP upregulated PKMYT1 expression and YAP prompted PKMYT1 transcriptional expression via TEAD1‐mediated direct binding to PKMYT1 promotor. Collectively, these findings suggest that PKMYT1, functioning as a direct gene target regulated by YAP/TEAD1, could serve as a potential indicator of progression and prognosis in BLCA. Further, PKMYT1 could serve as a novel therapeutic target for BLCA.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.