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MHY1485 mTOR activator

Cat.No.S7811

MHY1485 is a potent, and cell-permeable mTOR activator, and this compound also potently inhibits autophagy.

Chemical Structure

Molecular Weight: 387.39

Jump to Mechanism of Action Cell Culture & Working Concentration Solubility Chemical Information, Storage & Stability Specificity

Quality Control

Batch: Purity: 99.97%

99.97

Related Targets	PI3K Akt AMPK GSK-3 ATM/ATR DNA-PK PDPK1 PTEN PP2A PDK PI4K PIKfyve MELK
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Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Incubation Time	Activity Description	PMID
HT-29	Function assay	10 nM, 100 nM, 1 μM, 10 μM	48 h	significantly increased the expression of miR-212 in a dose-dependent manner	30021100
RSC96	Function assay	10 μM	1 day or 2 days	MHY1485 increased phospho-mTOR (Ser 2448), phosphoS6K1 (Thr 389) and NGF expression in RSC96 cells.	30229399
H9C2	Function assay	5 μM		MHY1485 abrogated the effects of Tanshinone IIA on LC3	30798134
Click to View More Cell Line Experimental Data

Chemical Information, Storage & Stability

Molecular Weight	387.39	Formula	C₁₇H₂₁N₇O₄	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	326914-06-1	Download SDF		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
Synonyms	N/A			Smiles	C1COCCN1C2=NC(=NC(=N2)NC3=CC=C(C=C3)[N+](=O)[O-])N4CCOCC4

Solubility

In vitro
Batch:

DMSO : 10 mg/mL (25.81 mM) Warmed with 50°C water bath; Ultrasonicated;
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
Mass value Mass unit	Concentration value Concentration unit	Volume value Volume unit	Molecular weight (g/mol)

Dilution Calculator Molecular Weight Calculator

In vivo batch selection In vivo Batch:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	0.800mg/ml (2.07mM)	Taking the 1 mL working solution as an example, add 50 μL of 16 mg/ml clarified DMSO stock solution to 400 μL PEG300, mix evenly to clarify it; add 50 μL Tween80 to the above system, mix evenly to clarify it; then continue to add 500 μL ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

Dosage: mg/kg

Average weight of animals: g

Dosing volume per animal: μL

Number of animals:

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO

% Tween 80

% ddH₂O

% DMSO

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Mechanism of Action

Targets/IC₅₀/Ki	mTOR ^[1]
In vitro	MHY1485 suppresses the basal autophagic flux. This compound causes the accumulation of LC3II and enlargement of the autophagosomes in a dose- and time- dependent manner.^[1] It increases phospho-mTOR levels and phosphorylation of downstream S6K1 and rpS6 proteins without affecting total mTOR content, total S6K1 and rpS6 levels. Short-term treatment of ovaries with this chemical followed by allo-transplantation promoted secondary follicle growth. Treatment with it and subsequent grafting allowed the derivation of mature oocytes and healthy offspring.^[2]
Kinase Assay	Activation of mTOR
Kinase Assay	Western blot analysis is performed to detect the change of total protein level and levels of phosphorylated forms of mTOR and 4E-BP1 reflecting the activity of mTOR. Ac2F cells are treated with MHY1485 of different concentrations and rapamycin 5 mM as a positive control for 1 h. Cells are washed with cold PBS and harvested. Cell lysates are prepared using RIPA buffer. Protein concentration is determined by the bicinchoninic acid (BCA) method. Equal amounts of protein are separated on 10-12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels. The gels are subsequently transferred onto a polyvinylidene difluoride membrane by electroblotting for 2 h at 60-75 V. The membranes are blocked in a 5% nonfat milk solution in Tris-buffered saline (TBS) with 0.5% Tween-20, and incubated with primary antibodies. Pre-stained protein markers are used for molecular-weight determination.
In vivo	MHY1485 is a potent, and cell-permeable mTORC1 agonist.
References	[1] https://pubmed.ncbi.nlm.nih.gov/22927967/ [2] https://pubmed.ncbi.nlm.nih.gov/25710488/ [3] https://pubmed.ncbi.nlm.nih.gov/36438491/ [4] https://pubmed.ncbi.nlm.nih.gov/34580888/

Applications

Methods	Biomarkers	Images	PMID
Western blot	p-mTOR / mTOR / p-AKT / AKT / p-S6K1 / S6K1 p-Nrf2 / Nrf2		28061443

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