WNK2-IN-1

WNK2-IN-1 (compound M04) (0.1-1000 μM; 24 h) exhibits an IC₅₀ of 416 μM for cell viability in human T/C-28a2 chondrocytes^[1].
WNK2-IN-1 (1-400 μM) reduces pSPAK levels in T/C-28a2 chondrocytes in a dose-dependent manner; at 200-400 μM, its reducing effect on pSPAK is comparable to that of the pan-WNK inhibitor WNK463 (HY-100626)^[1].
WNK2-IN-1 (1-200 μM) reduces WNK2 kinase activity in a dose-dependent manner in recombinant kinase assays; it does not affect WNK1 under the same conditions, and decreases WNK3 and WNK4 activity in a dose-dependent manner. However, the degree of inhibition of WNK3/WNK4 is weaker than that of WNK2, and does not reach the inhibitory level of WNK463^[1].
WNK2-IN-1 (400 μM; 5 min) reduces pSPAK to levels comparable to those achieved with WNK463 in T/C-28a2 chondrocytes; (R)-WNK2-IN-1 produces a similar reduction in pSPAK, whereas (S)-WNK2-IN-1 exhibits only a minor effect, indicating that the R-enantiomer is the active form^[1].
Compared with WNK463 (10 μM), WNK2-IN-1 (100 μM; 24 h) upregulates MMP2 and MMP13 and downregulates GREM2 and BMP4 in primary human chondrocytes not stimulated with IL1β; meanwhile, it enhances cell growth, Hedgehog signaling pathway inhibition, and immune response-related pathways, and downregulates autophagy-, starvation response-, and chemical stress-related pathways^[1].
WNK2-IN-1 (100 μM; 24 h; 10 ng/mL IL1β; 100 mM Sorbitol (HY-B0400)) reduces the expression of IL6 and CCL2 to control levels, increases COL2A1 expression, and prevents IL1β-induced downregulation of ACAN in T/C-28a2 chondrocytes^[1].
WNK2-IN-1 (100 μM; 24 h; 10 ng/mL IL1β; 100 mM Sorbitol) reduces the expression of CCL2 and FGF1 in primary human chondrocytes; no significant changes in gene expression are observed in control chondrocytes not stimulated with IL1β^[1].
Compared with WNK463 (10 μM), WNK2-IN-1 (100 μM; 24 h; 10 ng/mL IL1β) reduces the expression of BMP2, BMP4, NFKB1, MMP13 and FN1 and upregulates SLC1A5 and GREM1; meanwhile, it enhances pathways related to growth, TGFβ signaling, circadian rhythm and potassium transport, and downregulates pathways associated with TLR, interferon α/β/γ, interleukin, and RIPK-mediated NFκB activation^[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.