STO-609

Catalog No.S8274 Batch:S827402

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Technical Data

Formula

C19H10N2O3

Molecular Weight 314.29 CAS No. 52029-86-4
Solubility (25°C)* In vitro DMSO 3 mg/mL (9.54 mM)
Water Insoluble
Ethanol Insoluble
In vivo (Add solvents to the product individually and in order)
Clear solution
5% DMSO 95% Corn oil
0.075mg/ml Taking the 1 mL working solution as an example, add 50 μL of 1.5 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results. 
Clear solution
5%DMSO 40%PEG300 5%Tween80 50%ddH2O
0.15mg/ml Taking the 1 mL working solution as an example, add 50 μL of 3 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results. 
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description STO-609 is a specific inhibitor of the Ca2+/Calmodulin-dependent protein kinase kinase(CaM-KK) that inhibits the activities of recombinant CaM-KKα and CaM-KKβ isoforms, with Ki values of 80 and 15 ng/ml, respectively, and also inhibits their autophosphorylation activities. STO-609 inhibits AMPKK activity and inhibits autophagy.
Targets
AMPK [4] CaM-KKβ [1]
(Cell-free assay)
CaM-KKα [1]
(Cell-free assay)
47 nM(Ki) 0.25 μM(Ki)
In vitro

STO-609 inhibits the activities of recombinant CaM-KKα and CaM-KKβ isoforms, with Ki values of 80 and 15 ng/ml, respectively, and also inhibits their autophosphorylation activities. STO-609 is highly selective for CaM-KK without any significant effect on the downstream CaM kinases (CaM-KI and -IV). In transfected HeLa cells, STO-609 suppresses the Ca2+-induced activation of CaM-KIV in a dose-dependent manner. STO-609 can permeate cells and is a competitive inhibitor of ATP[1].

In vivo

In vivo administration of STO-609 results in increased osteoblasts and diminished osteoclasts, conferring significant protection from ovariectomy (OVX)-induced osteoporosis in adult mice[2]. ICV administration of STO-609 in vivo did not affect the counterregulatory responses to neuroglucopenia and AMPK activation induced by glucopenia[3].

Protocol (from reference)

Cell Assay:

[1]

  • Cell lines

    HeLa cells

  • Concentrations

    0.01-10 μg/ml

  • Incubation Time

    6 h

  • Method

    HeLa cells were maintained in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum. Cells were subcultured in 6-cm dishes 12 h before transfection. The cells were then transferred to serum-free medium and treated with a mixture of either 3 g of pME18s plasmid DNA or 3 g of HA(hemagglutinin-tagged)-CaM-KIV and 20 μg of LipofectAMINE reagent in 2.5 ml of medium. After 20 h of incubation, the cells were further cultured in serum-free medium for 6 h in either the absence or presence of various concentrations of STO-609 (0.01-10 μg/ml in Me2SO at a final concentration of 0.5%) and then treated with or without 1 μM ionomycin for 5 min. Stimulation was terminated by the addition of 1 ml of lysis buffer, 2 mM EDTA, 2 mM EGTA, 1% Nonidet P-40, 10% glycerol, 0.2 mM phenylmethylsulfonyl fluoride, 10 mg/liter leupeptin, 10 mg/liter trypsin inhibitor, and 1 μM microcystin LR), and the cells were lysed for 30min on ice. The cell extract was collected and centrifuged at 15,000×g for 15 min, the supernatant was precleared with 40 μl of Protein G-Sepharose for 2h at 4 °C, and the supernatant was mixed with 4 g of anti-HA antibody for 3h. 40 μl of Protein G-Sepharose was then applied to the extract and incubated overnight. The immunoprecipitated resin was washed three times with 1 ml of the lysis buffer and then washed with 1 ml of kinase buffer. Protein G-Sepharose with immunoprecipitated HA-CaM-KIV was subjected to the protein kinase assay in the presence of 1 mM EGTA using syntide-2 as a substrate.

Animal Study:

[2]

  • Animal Models

    WT, Camkk2−/− and Camk4−/− mice (all in C57BL/6 background)

  • Dosages

    10 μmol/kg

  • Administration

    i.p.

Selleck's STO-609 has been cited by 20 publications

Mechanical regulation of lipid and sugar absorption by Piezo1 in enterocytes [ Acta Pharm Sin B, 2024, 14(8):3576-3590] PubMed: 39220873
Targeting Dlat-Trpv3 pathway by hyperforin elicits non-canonical promotion of adipose thermogenesis as an effective anti-obesity strategy [ J Adv Res, 2024, S2090-1232(24)00555-1] PubMed: 39631519
Artemisinin conferred cytoprotection to human retinal pigment epithelial cells exposed to amiodarone-induced oxidative insult by activating the CaMKK2/AMPK/Nrf2 pathway [ J Transl Med, 2024, 22(1):844] PubMed: 39285426
Aβ25-35-induced autophagy and apoptosis are prevented by the CRMP2-derived peptide ST2-104 (R9-CBD3) via a CaMKKβ/AMPK/mTOR signaling hub [ PLoS One, 2024, 19(9):e0309794] PubMed: 39325788
Mechanical activation of VE-cadherin stimulates AMPK to increase endothelial cell metabolism and vasodilation [ bioRxiv, 2024, 2024.05.09.593171] PubMed: 38798670
Highly expressed SERCA2 triggers tumor cell autophagy and is a druggable vulnerability in triple-negative breast cancer [ Acta Pharm Sin B, 2022, 12(12):4407-4423] PubMed: 36561988
Amino acid starvation-induced LDLR trafficking accelerates lipoprotein endocytosis and LDL clearance [ EMBO Rep, 2022, e53373] PubMed: 34994492
Nuclear UHRF1 is a gate-keeper of cellular AMPK activity and function [ Cell Res, 2021, 10.1038/s41422-021-00565-y] PubMed: 34561619
Vitamin D-VDR (vitamin D receptor) regulates defective autophagy in renal tubular epithelial cell in streptozotocin-induced diabetic mice via the AMPK pathway [ Autophagy, 2021, 1-14] PubMed: 34432556
Elaiophylin reduces body weight and lowers glucose levels in obese mice by activating AMPK [ Cell Death Dis, 2021, 12(11):972] PubMed: 34671010

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

NOT FOR HUMAN, VETERINARY DIAGNOSTIC OR THERAPEUTIC USE.