MRT67307 HCl

Catalog No.S7948 Batch:S794801

Technical Data

Formula

C₂₆H₃₆N₆O₂· xHCl

Molecular Weight

464.6 (free-base)

CAS No.

1781882-89-0

Solubility (25°C)*

In vitro

DMSO

92 mg/mL

Water

92 mg/mL

Ethanol

92 mg/mL

In vivo (Add solvents to the product individually and in order)

Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O	4.6mg/ml	Taking the 1 mL working solution as an example, add 50 μL of 92 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.
Clear solution	5% DMSO 1 95% Corn oil	0.77mg/ml	Taking the 1 mL working solution as an example, add 50 μL of 15.4 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.

* <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

MRT67307 is a potent and dual IKKϵ and TBK1 inhibitor with IC50 of 160 and 19 nM, respectively. MRT67307 potently inhibits ULK1 and ULK2 and blocks autophagy.

Targets

IKKϵ ^[1] (Cell-free assay)	TBK1 ^[1] (Cell-free assay)
160 nM	190 nM

In vitro

In macrophages, MRT67307 prevents the phosphorylation of IRF3 and the production of IFNβ. In wild-type MEFs, MRT67307 enhances the IL-1-stimulated phosphorylation of p105 at Ser933 and RelA at both Ser468 and Ser536, and also enhances IL-1-stimulated activation of NF-κB-dependent gene transcription. ^[1] MRT67307 increases IL-10 production and suppresses proinflammatory cytokine production via a cAMP response element-Binding protein (CREB)-regulated transcriptional coactivator (CRTC) 3 Dependent Mechanism. ^[2] In addition, MRT67307 inhibit ULK and block autophagy in MEF cells. ^[3]

Protocol (from reference)

Kinase Assay:^{^[1]}	Protein kinase assays Substrates and kinases are diluted in 50 mM Tris/HCl (pH 7.5), 0.1% 2-mercaptoethanol, 0.1 mM EGTA and 10 mM magnesium acetate. Reactions are initiated with [γ-³²P]ATP (2500 c.p.m./pmol) to a final concentration of 0.1 mM and terminated after 15 min at 30 °C by the addition of SDS and EDTA (pH 7.0) to final concentrations of 1.0% (w/v) and 20 mM respectively. After heating for 5 min at 100 °C and separation by SDS/PAGE, the phosphorylated proteins are detected by autoradiography.
Cell Assay:^[3]	Cell lines MEFs and 293T cells Concentrations 10 μM Incubation Time 1 h Method MEFs and 293T cells were grown in DMEM, supplemented with 10% fetal bovine serum and penicillin/streptomycin, and cultured at 37 °C, 5% CO2. For induction of autophagy, cells were typically grown to 75% confluency, washed twice, and incubated in Earle's balanced salt solution (EBSS) for 1 h (or complete medium as a control) in the presence or absence of 10 μM MRT67307, 1 μM MRT68921, or 50 nM bafilomycin A1 and followed by lysis and immunoblotting with the indicated antibodies.

Kinase Assay:^{^[1]}

Protein kinase assays
Substrates and kinases are diluted in 50 mM Tris/HCl (pH 7.5), 0.1% 2-mercaptoethanol, 0.1 mM EGTA and 10 mM magnesium acetate. Reactions are initiated with [γ-³²P]ATP (2500 c.p.m./pmol) to a final concentration of 0.1 mM and terminated after 15 min at 30 °C by the addition of SDS and EDTA (pH 7.0) to final concentrations of 1.0% (w/v) and 20 mM respectively. After heating for 5 min at 100 °C and separation by SDS/PAGE, the phosphorylated proteins are detected by autoradiography.

Cell Assay:^[3]

Cell lines
MEFs and 293T cells
Concentrations
10 μM
Incubation Time
1 h
Method
MEFs and 293T cells were grown in DMEM, supplemented with 10% fetal bovine serum and penicillin/streptomycin, and cultured at 37 °C, 5% CO2. For induction of autophagy, cells were typically grown to 75% confluency, washed twice, and incubated in Earle's balanced salt solution (EBSS) for 1 h (or complete medium as a control) in the presence or absence of 10 μM MRT67307, 1 μM MRT68921, or 50 nM bafilomycin A1 and followed by lysis and immunoblotting with the indicated antibodies.

References

Customer Product Validation

: , , Methods Enzymol. 2017, 587:391-404

: Data from [Data independently produced by , , Mol Med Rep, 2018, 17(2):3085-3091]

Selleck's MRT67307 HCl has been cited by 17 publications

TBK1-Zyxin signaling controls tumor-associated macrophage recruitment to mitigate antitumor immunity [ EMBO J, 2024, 10.1038/s44318-024-00244-9]	PubMed: 39304793
mRNAs encoding self-DNA reactive cGAS enhance the immunogenicity of lipid nanoparticle vaccines [ mBio, 2023, 10.1128/mbio.02506-23]	PubMed: 37937842
A Chemical Proteomics Approach to Discover Regulators of Innate Immune Signaling [ Viruses, 2023, 15(5)1112]	PubMed: 37243198
Thymosin α-1 Reverses M2 Polarization of Tumor-Associated Macrophages during Efferocytosis [ Cancer Res, 2022, 82(10):1991-2002]	PubMed: 35364609
Revisiting the Mongolian Gerbil Model for Hepatitis E Virus by Reverse Genetics [ Microbiol Spectr, 2022, e0219321]	PubMed: 35230152
Human TBK1 deficiency leads to autoinflammation driven by TNF-induced cell death [ Cell, 2021, S0092-8674(21)00885-0]	PubMed: 34363755
Type I Interferon Regulates a Coordinated Gene Network to Enhance Cytotoxic T Cell-Mediated Tumor Killing. [ Cancer Discov, 2020, 10(3):382-393]	PubMed: 31974171
Deubiquitinase USP35 restrains STING-mediated interferon signaling in ovarian cancer [ Cell Death Differ, 2020, 10.1038/s41418-020-0588-y]	PubMed: 32678307
HSV-1 and Zika Virus but Not SARS-CoV-2 Replicate in the Human Cornea and Are Restricted by Corneal Type III Interferon [ Cell Rep, 2020, 33(5):108339]	PubMed: 33147451
SARS-CoV-2 Infects Human Engineered Heart Tissues and Models COVID-19 Myocarditis [ bioRxiv, 2020, 2020.11.04.364315]	PubMed: 33173875

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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.