STO-609 suppresses the functionality of both recombinant CaM-KKα and CaM-KKβ isoforms, exhibiting Ki measurements of 80 and 15 ng/mL for each, respectively, and hampers their self-phosphorylation processes. It demonstrates a pronounced selectivity towards CaM-KK, showing negligible influence on the subsequent CaM kinases (CaM-KI and -IV), with the IC50 against CaM-KII being 10 μg/mL. Both the constitutively active and the wild-type variants of CaM-KKα are inhibited by STO-609. It also curtails the Ca2+-prompted activation of CaM-KIV within HeLa cells that have undergone transfection, operating in a concentration-dependent fashion. Moreover, at a dose of 1μg/mL, STO-609 markedly lowers the inherent activity of CaM-KK in SH-SY5Y neuroblastoma cells, achieving an inhibition rate of 80% [1].
体外研究
STO-609 suppresses the functionality of both recombinant CaM-KKα and CaM-KKβ isoforms, exhibiting Ki measurements of 80 and 15 ng/mL for each, respectively, and hampers their self-phosphorylation processes. It demonstrates a pronounced selectivity towards CaM-KK, showing negligible influence on the subsequent CaM kinases (CaM-KI and -IV), with the IC50 against CaM-KII being 10 μg/mL. Both the constitutively active and the wild-type variants of CaM-KKα are inhibited by STO-609. It also curtails the Ca2+-prompted activation of CaM-KIV within HeLa cells that have undergone transfection, operating in a concentration-dependent fashion. Moreover, at a dose of 1μg/mL, STO-609 markedly lowers the inherent activity of CaM-KK in SH-SY5Y neuroblastoma cells, achieving an inhibition rate of 80% [1].
作用机制
STO-609 permeates cells and competes with ATP to bind to the ATP-binding pocket on CaM-KK, thereby preventing them from activating the phosphorylation of CaM kinases.
STO-609 细胞实验
Cell Line
Concentration
Treated Time
Description
References
LNCaP cells
5 µM
24 h
Inhibited AR upregulation induced by SDHA/SDHB subunit silencing
EMBO Mol Med. 2021 May 7;13(5):e13427.
A549 cells
1, 2 or 5 µg ml−1
6 h
To examine the effect of STO-609 on AMPK activity, results showed that STO-609 inhibited AMPK activity
Nature. 2012 May 9;485(7400):661-5.
Apc10.1 cells
0.01-10 μM
6 days
To evaluate the effect of STO-609 on AMPK signaling, results showed that low concentrations of STO-609 activated AMPK and induced autophagy and senescence.
Sci Transl Med. 2015 Jul 29;7(298):298ra117.
3T3-L1 cells
10 μg/mL
1 h
Inhibition of Ca2+/Calmodulin-dependent protein kinase kinase (CaMKK) activity
Mol Cell. 2020 Sep 17;79(6):934-949.e14.
HME cells
10 µM
48 h
STO-609 caused cell cycle arrest and cell death in MYC and T58A HME cells.
Br J Cancer. 2020 Mar;122(6):868-884.
SUM159PT cells
10 µM
2 h
STO-609 decreased p-AMPK levels, indicating that AMPK activation is dependent on CAMKK2.
Br J Cancer. 2020 Mar;122(6):868-884.
STO-609 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Mice
Pten null prostate cancer model
Subcutaneous osmotic pump
15 mg/ml
6 weeks
STO-609 treatment significantly reduced prostate weight and slowed prostate cancer progression.
Cancer Res. 2018 Dec 15;78(24):6747-6761
Mice
Breast cancer model
Intraperitoneal injection
100 μM/kg
Three times per week until the end of the experiment
STO-609 inhibited CaMKK2, attenuated mammary tumor growth, and promoted the accumulation of CD8+ T cells and NK1.1+ T cells within the tumors.
Nat Commun. 2019 Jun 4;10(1):2450
Mice
Destabilization of the medial meniscus (DMM) model
Intraperitoneal injection
0.033 mg/kg
Three times per week, for 8 or 12 weeks
Inhibition or absence of CaMKK2 protected against DMM-associated destruction of the cartilage, subchondral bone alterations and synovial inflammation
Osteoarthritis Cartilage. 2022 Jan;30(1):124-136
Mice
Non-alcoholic fatty liver disease (NAFLD) models
Intraperitoneal injection
30 μM/kg
Once daily for 4 weeks
To evaluate the therapeutic effect of STO-609 on NAFLD, results showed that STO-609 significantly improved hepatic steatosis
Sci Rep. 2017 Sep 18;7(1):11793
Mice
LNCaP xenograft model
Oral
10 mg/kg
Three times a week for 12 weeks
IVM treatment significantly reduced tumor growth and PSA levels, and inhibited the activity of the p-CAMKK2/p-p38/p-Hsp27 axis
EMBO Mol Med. 2021 May 7;13(5):e13427.
Mice
High-fat diet model
Oral
2.1 μg
Single dose
To evaluate the effect of STO-609 on AMPK signaling, results showed that STO-609 activated AMPK and induced autophagy and senescence within 30 minutes.
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