MHY1485

Catalog No.S7811 Batch:S781103

Technical Data

Formula

C₁₇H₂₁N₇O₄

Molecular Weight

387.39

CAS No.

326914-06-1

Solubility (25°C)*

In vitro

DMSO

10 mg/mL (25.81 mM)

Water

Insoluble

Ethanol

Insoluble

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

Clear solution

5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O

0.8mg/ml

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.

* <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

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

Targets

mTOR ^[1]

In vitro

MHY1485 suppresses the basal autophagic flux. MHY1485 causes the accumulation of LC3II and enlargement of the autophagosomes in a dose- and time- dependent manner.^[1]

MHY1485 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 MHY1485 followed by allo-transplantation promoted secondary follicle growth. Treatment with MHY1485 and subsequent grafting allowed the derivation of mature oocytes and healthy offspring.^[2]

In vivo

MHY1485 is a potent, and cell-permeable mTORC1 agonist.

Protocol (from reference)

Kinase Assay:^{^[1]}	Activation of mTOR 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 (50 mM Tris-HCl, 150 mM NaCl, 1% NP-40, 1 mM DTT, 0.1 mM NaF, 1 mM PMSF, 1 mg/mL pepstatin, 1 mg/mL leupeptin, and 1 mg/mL aprotinin). 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.
Cell Assay:^{^[3]}	Cell lines TU177 cells Concentrations 10 µM Incubation Time 24 h Method Cells were treated with indicated concentration of drug for 24 h.

Kinase Assay:^{^[1]}

Activation of mTOR
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 (50 mM Tris-HCl, 150 mM NaCl, 1% NP-40, 1 mM DTT, 0.1 mM NaF, 1 mM PMSF, 1 mg/mL pepstatin, 1 mg/mL leupeptin, and 1 mg/mL aprotinin). 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.

Cell Assay:^{^[3]}

Cell lines
TU177 cells
Concentrations
10 µM
Incubation Time
24 h
Method
Cells were treated with indicated concentration of drug for 24 h.

References

[4] Fang Y, et al. Hepatology. 2022 Jun;75(6):1386-1401.

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Customer Product Validation

: Data from [Data independently produced by , , FASEB J, 2018, 32(1):512-528]

: Data from [Data independently produced by , , Int J Biol Sci, 2017, 13(11):1398-1408]

: Data from [Data independently produced by , , Biochem J, 2017, 474(16):2733-2747]

: Data from [Data independently produced by , , Oncol Rep, 2018, 40(1):425-433]

Selleck's MHY1485 has been cited by 80 publications

Gestational exposure to PM2.5 disrupts fetal development by suppressing placental trophoblast syncytialization via progranulin/mTOR signaling [ Sci Total Environ, 2024, 921:171101]	PubMed: 38387595
Metformin improves memory via AMPK/mTOR-dependent route in a rat model of Alzheimer's disease [ Iran J Basic Med Sci, 2024, 27(3):360-365.]	PubMed: 38333746
MHY1485 potentiates immunogenic cell death induction and anti-cancer immunity following irradiation [ J Radiat Res, 2024, rrad107]	PubMed: 38330507
Antitumor activity of afatinib in EGFR T790M-negative human oral cancer therapeutically targets mTOR/Mcl-1 signaling axis [ Research Square, 2024, 10.21203/rs.3.rs-3872267/v1]	PubMed: none
Cold exposure protects against medial arterial calcification development via autophagy [ J Nanobiotechnology, 2023, 21(1):226]	PubMed: 37461031
Butyrate dictates ferroptosis sensitivity through FFAR2-mTOR signaling [ Cell Death Dis, 2023, 14(4):292]	PubMed: 37185889
A novel gene REPTOR2 activates the autophagic degradation of wing disc in pea aphid [ Elife, 2023, 12e83023]	PubMed: 36943031
A novel gene REPTOR2 activates the autophagic degradation of wing disc in pea aphid [ Elife, 2023, 12e83023]	PubMed: 36943031
Propionate promotes gluconeogenesis by regulating mechanistic target of rapamycin (mTOR) pathway in calf hepatocytes [ Anim Nutr, 2023, 10.1016/j.aninu.2023.07.001]	PubMed: 37841648
A unique poly(A) tail profile uncovers the stability and translational activation of TOP transcripts during neuronal differentiation [ iScience, 2023, 26(9):107511]	PubMed: 37636056

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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.

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