The With-No-Lysine (K) (WNK) kinases play an essential role in regulating blood pressure and homeostasis of body fluid and electrolyte. WNK463 is an orally bioavailable pan-WNK-kinase inhibitor that inhibits the in vitro kinase activity of all four WNK family members (WNK1, WNK2, WNK3, and WNK4) with IC50 values of 5nM, 1nM, 6nM, and 9nM, respectively. It also inhibits WNK1-catalyzed phosphorylation of the native WNK substrate, oxidative stress response 1, with an IC50 value of 41nM. In spontaneously hypertensive rats (34 – 42 weeks of age), oral administration of WNK463 at doses of 1, 3, or 10mg/kg body weight resulted in a dose-dependent decrease in blood pressure, a simultaneous increase in heart rate, and significant increases in urine output, urinary sodium, and potassium excretion rate[1]
WNK463 细胞实验
Cell Line
Concentration
Treated Time
Description
References
H2009
1 µM
48 h
WNK463 significantly increased the death of cancer cells by activated PBMCs
Exp Mol Med. 2022 Nov;54(11):1913-1926.
MC38
1 µM
72 h
WNK463 reduced PD-L1 expression in MC38 cells
Exp Mol Med. 2022 Nov;54(11):1913-1926.
CD4+ T cells
2.5 μM
1 h
To investigate the role of WNK1 kinase activity in TCR/CD28-induced proliferation of CD4+ T cells, results showed that WNK1 kinase activity is required for TCR/CD28-induced proliferation of CD4+ T cells.
Nat Commun. 2025 Feb 21;16(1):1857.
LR73 cells
10 μM
1 h
To evaluate the effect of WNK463 on the phagocytosis of apoptotic cells by LR73 cells, results showed that WNK463 treatment enhanced efferocytosis.
Nat Cell Biol. 2019 Dec;21(12):1532-1543.
bone marrow-derived macrophages
10 μM
1 h
To evaluate the effect of WNK463 on the phagocytosis of apoptotic cells by bone marrow-derived macrophages, results showed that WNK463 treatment enhanced efferocytosis.
Nat Cell Biol. 2019 Dec;21(12):1532-1543.
peritoneal macrophages
10 μM
1 h
To evaluate the effect of WNK463 on the phagocytosis of apoptotic cells by peritoneal macrophages, results showed that WNK463 treatment enhanced efferocytosis.
Nat Cell Biol. 2019 Dec;21(12):1532-1543.
mouse podocytes
0.5 to 20 μM
WNK463 reduced phosphorylation of both SPAK and OSR1, indicating that OSR1/SPAK are partially activated by WNK prior to osmotic stimulation.
Front Cell Dev Biol. 2021 Feb 2;8:618898.
HEK293T cells
10 μM
2 h
Inhibition of WNK kinases, enhancing the catalytic activity of Nedd4-2
Proc Natl Acad Sci U S A. 2022 Jul 26;119(30):e2122495119.
MDCK cells
10 μM
2 h
Inhibition of WNK kinases, enhancing the catalytic activity of Nedd4-2
Proc Natl Acad Sci U S A. 2022 Jul 26;119(30):e2122495119.
mpkCCD c14 cells
100 nM
24 h
To investigate the effect of WNK463 on TRPV4 activity, results showed that WNK463 significantly reduced TRPV4-dependent Ca2+ influx.
Cells. 2021 Jun 12;10(6):1482.
MDA-MB-231 cells
1 μM
24 h
WNK463 reduced invasion of MDA-MB-231 cells in collagen matrices
Mol Cancer Ther. 2021 Oct;20(10):1800-1808.
WNK463 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Mice
RAG1-deficient mice
Intraperitoneal injection
2 mg
3 consecutive days
To investigate the role of WNK1 in T cells, results showed that WNK1-deficient T cells are unable to support a TD antibody response.
Nat Commun. 2025 Feb 21;16(1):1857.
Mice
Thymic engulfment model and peritoneal engulfment model
Intraperitoneal injection
1 mg/kg
Injected 1 hour before and concurrently with injection of apoptotic cells or dexamethasone, lasting for 12 hours
To evaluate the effect of WNK463 on the phagocytosis of apoptotic cells by resident macrophages in mice, results showed that WNK463 treatment enhanced efferocytosis.
Nat Cell Biol. 2019 Dec;21(12):1532-1543.
Mice
C57BL/6 mice
Drinking water
1 mg/kg
Once daily for 3 days
To investigate the effect of WNK463 on TRPV4 activity, results showed that WNK463 significantly reduced TRPV4-dependent Ca2+ influx.
Cells. 2021 Jun 12;10(6):1482.
Mice
MDA-MB-231 bone metastasis model
Oral
1-1.5 mg/kg
Daily administration for 4 weeks
WNK463 reduced tumor burden and metastasis in mice
Mol Cancer Ther. 2021 Oct;20(10):1800-1808.
Mice
Model of ischemic stroke
Intraperitoneal injection
5 mg/kg
Two doses at 3 h and 8 h post reperfusion
ZT-1a reduced brain infarct volume and cerebral edema, and improved neurological function
Nat Commun. 2020 Jan 7;11(1):78.
Mice
MC38 syngeneic colon adenocarcinoma model
Oral
5 or 10 mg/kg
Once daily for 9 days
WNK463 dose-dependently suppressed tumor growth and increased IFN-γ and TNF-α produced by CD4+ and CD8+ T cells in tumor-infiltrating lymphocytes
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