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Chemical Structure| 2012607-27-9 Chemical Structure| 2012607-27-9

Structure of WNK463
CAS No.: 2012607-27-9

Chemical Structure| 2012607-27-9

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WNK463 is a potant With-No-Lysine (WNK) kinase inhibitor with IC50s of 5/1/6/9 nM for WNK1/2/3/4.

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Product Details of WNK463

CAS No. :2012607-27-9
Formula : C21H24F3N7O2
M.W : 463.46
SMILES Code : O=C(C1=CN=CN1C2CCN(C3=NC=C(C4=NN=C(C(F)(F)F)O4)C=C3)CC2)NC(C)(C)C
MDL No. :MFCD30489744
InChI Key :HWSHOMMVLGBIDN-UHFFFAOYSA-N
Pubchem ID :121487936

Safety of WNK463

GHS Pictogram:
Signal Word:Warning
Hazard Statements:H302-H315-H319-H335
Precautionary Statements:P261-P305+P351+P338

Related Pathways of WNK463

GPCR

Isoform Comparison

Biological Activity

In Vitro:

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.

In Vivo:

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 Exp Mol Med. 2022 Nov;54(11):1913-1926.

Protocol

Bio Calculators
Preparing Stock Solutions 1mg 5mg 10mg

1 mM

5 mM

10 mM

2.16mL

0.43mL

0.22mL

10.79mL

2.16mL

1.08mL

21.58mL

4.32mL

2.16mL

Dissolving Methods
Please choose the appropriate dissolution scheme according to your animal administration guide.For the following dissolution schemes, clear stock solution should be prepared according to in vitro experiments, and then cosolvent should be added in turn:

in order to ensure the reliability of the experimental results, the clarified stock solution can be properly preserved according to the storage conditions; The working fluid for in vivo experiment is recommended to be prepared now and used on the same day;

The percentage shown in front of the following solvent refers to the volume ratio of the solvent in the final solution; If precipitation or precipitation occurs in the preparation process, it can be assisted by heating and/or ultrasound.
Protocol 1
Protocol 2

References

 

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