NUAK1-IN-3

NUAK1-IN-3 is a potent and selective inhibitor of NUAK1, with an IC50 of 0.49 nM. It also inhibits NUAK2 and JAK3, exhibiting IC50 values of 265 nM and 225 nM respectively. The compound interacts with NUAK1 at Glu139, forming a salt bridge with its bicyclic nitrogen atom and Asp142, and enhances hydrophobic interactions through a fluorine atom. NUAK1-IN-3 reduces the phosphorylation of MYPT1, thereby inhibiting the NUAK1-MYPT1 signaling axis while suppressing the proliferation, migration, and invasion of triple-negative breast cancer cells. It can reverse TGF-β1-induced epithelial-mesenchymal transition (EMT) marker changes, downregulating Snail and N-cadherin expression, while upregulating E-cadherin expression in tumor tissues. Additionally, NUAK1-IN-3 inhibits tumor growth in triple-negative breast cancer xenograft models, making it a useful tool for research related to this cancer.

NUAK1:0.49 nM

NUAK1-IN-3 (Compound 10i) is a potent inhibitor of NUAK1 with an IC₅₀ of 0.49 nM, demonstrating selectivity over JAK3 (IC₅₀ = 225 nM) and NUAK2 (IC₅₀ = 265 nM) at 459-fold and 541-fold, respectively. At concentrations of 0-50 μM for 96 hours, NUAK1-IN-3 inhibits the proliferation of triple-negative breast cancer cells BT549 and MDA-MB-231, with IC₅₀ values of 2.8 μM and 3.4 μM, respectively. Additionally, NUAK1-IN-3 at 1000-10000 nM for 16 hours reduces the phosphorylation of MYPT1 in a concentration-dependent manner in these cancer cells. When used in concentrations between 1-5 μM for 24 hours, it dose-dependently inhibits the migration of BT549 and MDA-MB-231 cells. At 3-10 μM over 24 hours, it also suppresses the invasion of these cells, with the maximum effect at 10 μM comparable to NUAK1 knockdown via siRNA. Furthermore, NUAK1-IN-3 in the range of 1-10 μM for 24 hours impedes the EMT pathway by downregulating N-cadherin and Snail expression in the presence of TGF-β₁ in a dose-dependent fashion in these cells.

NUAK1-IN-3 (Compound 10i) administered intraperitoneally at doses of 60-100 mg/kg once daily for 16 consecutive days effectively inhibits the growth of MDA-MB-231 triple-negative breast cancer xenografts in NCG mice in a dose-dependent manner.