Cat. No.: HY-125535

OSU-53

OSU-53 is an orally active AMPK activator and mTOR inhibitor, with an IC₅₀ of 0.3 μM for AMPK, an EC₅₀ of 2-5 μM for AMPK, and an IC₅₀ of 8.23 μM for mTOR. OSU-53 inhibits the Akt signaling pathway, directly activates AMPK by binding to its autoinhibitory domain, reduces IKKβ-mediated phosphorylation of Foxo3a, blocks Akt-mediated phosphorylation of MDM2, and promotes the nuclear localization and stabilization of Foxo3a, while directly inhibiting mTOR. OSU-53 inhibits epithelial-mesenchymal transition (EMT), induces epithelial phenotype, suppresses invasion and metastasis, induces autophagy, inhibits the activation of ERK and Akt, reduces the secretion of nitric oxide and proinflammatory cytokines, and inhibits the migration of MDSC; at high doses, it induces MDSC apoptosis, attenuates the immunosuppressive effect of MDSC, reduces MDSC levels, and blocks incision-induced mechanical hyperalgesia. OSU-53 can be used in studies related to breast cancer, prostate cancer, thyroid cancer, melanoma and postoperative pain.

For research use only. We do not sell to patients.

CAS No.: 1290069-19-0
Formula: C₂₅H₂₄F₃N₃O₆S₂
Molecular Weight:583.60

Storage:

Please store the product under the recommended conditions in the Certificate of Analysis.

Data Sheet (301 KB)

OSU-53 Chemical Structure

CAS No.: 1290069-19-0

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Use of MCE Products

Strong Fluorescence Signal / Multi-Disciplinary Applications / High Fluorescence Specificity

Biological Activity
- Description
- IC₅₀ & Target
- Cellular Effect
- In Vitro
- In Vivo
Chemical Information
Purity & Documentation
References
Calculators
Help & FAQs
Classifications
Bioz

Biological Activity

Description

IC₅₀ & Target^[1]^[2]^[3]^[4]

AMPK

0.3 μM (IC₅₀)

AMPK

2-5 μM (EC₅₀)

mTOR

8.23 μM (IC₅₀)

Akt

IKKβ

FOXO3a

MDM2

MDSC

All AMPK Isoforms :

All mTOR Isoforms :

All Akt Isoforms :

All IKK Isoforms :

All FOXO Isoforms :

FOXO1
FOXO3

All ERK Isoforms :

Cellular Effect

Cell Line	Type	Value	Description	References
MCF7	IC50	5 μM	Antiproliferative activity against human MCF-7 cancer cells assessed as reduction in cell viability incubated for 72 hrs by MTT assay. Antiproliferative activity against human MCF-7 cancer cells assessed as reduction in cell viability incubated for 72 hrs by MTT assay.	24994714
MDA-MB-231	IC50	7.5 μM	Antiproliferative activity against human MDA-MB-231 cancer cells assessed as reduction in cell viability incubated for 72 hrs by MTT assay. Antiproliferative activity against human MDA-MB-231 cancer cells assessed as reduction in cell viability incubated for 72 hrs by MTT assay.	24994714
MDA-MB-468	IC50	5 μM	Antiproliferative activity against human MDA-MB-468 cancer cells assessed as reduction in cell viability incubated for 72 hrs by MTT assay. Antiproliferative activity against human MDA-MB-468 cancer cells assessed as reduction in cell viability incubated for 72 hrs by MTT assay.	24994714
4T1	IC50	6 μM	Antiproliferative activity against mouse 4T1 cancer cells assessed as reduction in cell viability incubated for 72 hrs by MTT assay. Antiproliferative activity against mouse 4T1 cancer cells assessed as reduction in cell viability incubated for 72 hrs by MTT assay.	24994714
LNCaP	IC50	9 μM	Antiproliferative activity against human LNCaP cancer cells assessed as reduction in cell viability incubated for 72 hrs by MTT assay. Antiproliferative activity against human LNCaP cancer cells assessed as reduction in cell viability incubated for 72 hrs by MTT assay.	24994714
DU-145	IC50	8 μM	Antiproliferative activity against human DU-145 cancer cells assessed as reduction in cell viability incubated for 72 hrs by MTT assay. Antiproliferative activity against human DU-145 cancer cells assessed as reduction in cell viability incubated for 72 hrs by MTT assay.	24994714
PC-3	IC50	5 μM	Antiproliferative activity against human PC-3 cancer cells assessed as reduction in cell viability incubated for 72 hrs by MTT assay. Antiproliferative activity against human PC-3 cancer cells assessed as reduction in cell viability incubated for 72 hrs by MTT assay.	24994714

In Vitro

OSU-53 directly activates recombinant AMPK α1β1γ2 protein with an IC₅₀ of 0.3 μM^[1].
OSU-53 (18 h) dose-dependently inhibits the migration of MDA-MB-231 and PC-3 cells, with effects independent of cell viability reduction^[1].
OSU-53 (24 h) dose-dependently inhibits the invasion of MDA-MB-231 and PC-3 cells, with effects independent of cell viability reduction^[1].
OSU-53 (9-16 days) dose-dependently inhibits the formation of invasive, stellate colonies by MDA-MB-231 and PC-3 cells in three-dimensional culture^[1].
OSU-53 reduces F-actin stress fiber formation in MDA-MB-231 cells^[1].
OSU-53 (1-5 μM; 18 h) activates AMPK in the murine MDSC cell line MSC-2, as shown by increased p-AMPK levels following 5 μM treatment for 18 hours, without changing total AMPK levels^[2].
OSU-53 (5-10 μM; 72 h) inhibits the viability of MCF-7, MDA-MB-231, MDA-MB-468, 4T1, LNCaP, DU-145, and PC-3 cancer cell lines with IC₅₀ values of 5 μM, 7.5 μM, 5 μM, 6 μM, 9 μM, 8 μM, and 5 μM respectively after 72 h of treatment, and has no antiproliferative effect on normal MECs and PrECs^[1].
OSU-53 (2.5-10 μM) reverses the mesenchymal phenotype of MDA-MB-231, 4T1, PC-3, and MDA-MB-468 cells via AMPK activation, inducing increased expression of epithelial markers and decreased expression of mesenchymal markers at the mRNA and/or protein level^[1].
OSU-53 (5 μM) induces Foxo3a binding to the FBE3 and FBE4 regions of the E-cadherin promoter in MDA-MB-468, PC-3, and MDA-MB-231 cells^[1].
OSU-53 (5 μM; 24 h) promotes nuclear localization of Foxo3a and cytoplasmic sequestration of MDM2 in MDA-MB-231, 4T1, and PC-3 cells by suppressing Akt and IKKβ phosphorylation^[1].
OSU-53 (5 μM; 12 h) increases Foxo3a protein stability by reducing MDM2-mediated ubiquitination in cancer cells, an effect blocked by ectopic MDM2 or constitutively active Akt expression^[1].
OSU-53 (1-10 μM; 24-72 h) inhibits the proliferation of all tested human thyroid cancer cell lines in a dose-dependent manner, with greatest potency (GI₅₀ 1.63-4.51 μM) in lines with activating RAS or BRAF^V600E mutations, and lower potency (GI₅₀ 5.90-7.26 μM) in FTC133 and TPC1 cells^[3].
OSU-53 (5 μM; 6-72 h) activates AMPK, suppresses mTOR signaling, and modulates ERK and AKT activation in human ATC cell lines, with cell-specific differences based on RAS or BRAF^V600E mutation status^[3].
OSU-53 (1-10 μM; 6-72 h) activates AMPK in BCPAP and FTC133 human DTC cell lines, suppresses mTOR signaling in all three tested DTC lines, and modulates ERK and AKT activation in a cell-specific manner^[3].
OSU-53 (1-10 μM; 24-72 h; 5 μM; 24-48 h) does not rely primarily on AMPK activation for induced growth inhibition or autophagy in BCPAP and FTC133 human DTC cell lines, though AMPK contributes partially to mTOR/p70S6K signaling suppression in these cells^[3].
OSU-53 (0.01-15 μM; cell-free assay; 1-10 μM; 24 h) directly inhibits mTOR activity in a cell-free system (IC₅₀ 8.23 μM) and reduces mTOR phosphorylation in U-Hth-7 and Hth-104 human thyroid cancer cell lines^[3].
OSU-53 (0.5-10 μM; 18 h, overnight) does not induce apoptosis in the murine MDSC cell line MSC-2 at concentrations of 0.5, 1, and 5 μM, but induces apoptosis at 10 μM^[2].
OSU-53 (0.5-5 μM; 12 h pre-incubation plus 24 h LPS stimulation) reduces nitric oxide production in LPS-stimulated murine MDSC cell line MSC-2, with a 3-fold decrease observed at 5 μM treatment^[2].
OSU-53 (0.5-5 μM; overnight) reduces migration of the murine MDSC cell line MSC-2 toward tumor cell-derived chemoattractants, with a significant decrease observed at 5 μM treatment^[2].
OSU-53 (0.5-5 μM; 24 h) reduces secretion of pro-inflammatory cytokines in LPS-stimulated murine MDSC cell line MSC-2, with a 1.5-fold decrease in TNF-α and 2.9-fold decrease in IL-6 observed at 5 μM treatment^[2].
OSU-53 (5 μM; 24-48 h) induces autophagy in human thyroid cancer cell lines with RAS or BRAF^V600E mutations, but does not induce meaningful apoptosis in any of the tested lines^[3].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Cell Viability Assay^[1]

Cell Line:

MCF-7, MDA-MB-231, MDA-MB-468, 4T1, LNCaP, DU-145, PC-3, normal mammary epithelial cells (MECs), normal prostate epithelial cells (PrECs)
Concentration:

5-10 μM
Incubation Time:

72 h
Result:

Inhibited viability of seven cancer cell lines with IC50 values of 5 μM, 7.5 μM, 5 μM, 6 μM, 9 μM, 8 μM, and 5 μM respectively after 72 h.
Exhibited no antiproliferative effect on normal MECs and PrECs.

Apoptosis Analysis^[2]

Cell Line:

murine MDSC cell line MSC-2
Concentration:

0.5 μM, 1 μM, 5 μM, 10 μM
Incubation Time:

18 h (0.5 μM, 1 μM, 5 μM); overnight (10 μM)
Result:

Did not induce apoptosis in MSC-2 cells at concentrations up to 5 μM.
Caused a significant increase in apoptotic cells at the 10 μM concentration.

Western Blot Analysis^[2]

Cell Line:

murine MDSC cell line MSC-2
Concentration:

1 μM, 5 μM
Incubation Time:

18 h
Result:

Did not alter total AMPK protein levels in MSC-2 cells.
Induced an increase in phosphorylated AMPK (p-AMPK) levels at 5 μM.

Cell Migration Assay ^[2]

Cell Line:

murine MDSC cell line MSC-2
Concentration:

0.5 μM, 1 μM, 5 μM
Incubation Time:

overnight
Result:

Caused a significant reduction in the number of MSC-2 cells migrating toward tumor cell supernatant at 5 μM compared to controls.

ELISA Assay^[2]

Cell Line:

murine MDSC cell line MSC-2
Concentration:

0.5 μM, 1 μM, 5 μM
Incubation Time:

24 h
Result:

Caused a 1.5-fold decrease in TNF-α levels at 5 μM in LPS-stimulated MSC-2 cells.
Caused a 2.9-fold decrease in IL-6 levels at 5 μM in LPS-stimulated MSC-2 cells.

Cell Proliferation Assay^[3]

Cell Line:

C643, U-Hth-7, SW1736, Hth-104, FTC133, TPC1, BCPAP
Concentration:

1 μM, 5 μM, 10 μM
Incubation Time:

24 h; 48 h; 72 h
Result:

Inhibited proliferation of all 7 thyroid cancer cell lines in a dose-dependent manner.
Reduced GI₅₀ values to 1.63-4.51 μM in cell lines with activating RAS or BRAF V600E mutations.
Resulted in a GI₅₀ of 5.90 μM in FTC133 (PTEN null) cells.
Resulted in a GI₅₀ of 7.26 μM in TPC1 (RET/PTC1) cells.
Caused significantly less growth inhibition in FTC133 and TPC1 compared to BCPAP at 5 μM.

Western Blot Analysis^[3]

Cell Line:

C643, U-Hth-7, SW1736, Hth-104
Concentration:

5 μM
Incubation Time:

6 h; 12 h; 24 h; 48 h; 72 h
Result:

Induced time-dependent activation of AMPK (increased p-AMPK relative to total AMPK) in all four ATC cell lines.
Reached peak AMPK activation at 12 hours and declined by 48 hours in RAS mutant lines (C643, U-Hth-7).
Activated TSC2 (increased p-TSC2) in three of the four cell lines, with no appreciable increase in C643.
Suppressed mTOR signaling (decreased p-p70S6K and p-S6) across all four cell lines.
Suppressed ERK activation (decreased p-ERK) after 48 hours in BRAF V600E mutant lines (SW1736, Hth-104) but not consistently in RAS mutant lines.
Down-regulated AKT activation (decreased p-AKT) in C643, U-Hth-7, and SW1736, but not Hth-104.

Western Blot Analysis^[3]

Cell Line:

BCPAP, FTC133, TPC1, Hth-104
Concentration:

5 μM; 1, 2.5, 5, 7.5, 10 μM
Incubation Time:

6 h; 12 h; 24 h; 48 h; 72 h (5 μM treatment); 24 h (Hth-104 dose-response); 48 h (FTC133, TPC1 dose-response)
Result:

Induced time-dependent AMPK activation (increased p-AMPK) in BCPAP and FTC133, but not TPC1, with 5 μM treatment.
Suppressed p-p70S6K activation in all three DTC cell lines, with complete abolition of p-p70S6K in BCPAP after 6 hours.
Induced time-dependent inactivation of ERK (decreased p-ERK) in FTC133, with variable inactivation in BCPAP and TPC1.
Induced time-dependent inactivation of AKT (decreased p-AKT) in FTC133 and TPC1, but not BCPAP.
Caused modest dose-dependent AMPK activation in Hth-104 and FTC133, but not TPC1, with dose-response treatment.
Decreased p-p70S6K in all three cell lines with dose-response treatment.

Western Blot Analysis^[3]

Cell Line:

C643, U-Hth-7, SW1736, Hth-104, BCPAP, FTC133, TPC1
Concentration:

5 μM
Incubation Time:

6 h; 12 h; 24 h; 48 h; 72 h
Result:

Induced increased conversion of LC3 I to LC3 II (autophagy marker) in all BRAF V600E mutant lines (BCPAP, Hth-104, SW1736) and RAS mutant lines (C643, U-Hth-7).
Caused modest LC3 I/II conversion in TPC1 and absent conversion in FTC133.
Induced a slight increase in PARP cleavage (apoptosis marker) only in U-Hth-7 after 48 hours of treatment.

In Vivo

OSU-53 (50-100 mg/kg; p.o.; daily; 21 days) activates AMPK, reverses the mesenchymal phenotype of 4T1 tumors, and reduces lung metastatic burden in syngeneic BALB/c mice^[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Model:

BALB/cAnNCr (female, 5-7 weeks of age, syngeneic orthotopic breast cancer model via injection of 4T1 cells)^[1]
Dosage:

50 mg/kg; 100 mg/kg
Administration:

p.o.; daily; 21 days
Result:

Exhibited no changes in body weight.
Modestly inhibited primary tumor growth.
Dose-dependently increased phosphorylation of AMPK and ACC, reduced phosphorylation of Akt, and increased Foxo3a expression in tumor lysates.
Increased E-cadherin levels and decreased vimentin, YB-1, Snail, and Twist levels in tumor lysates.
Reduced the number and size of metastatic nodules on the lung surface relative to controls.

Chemical Information

CAS No. 1290069-19-0
Molecular Weight 583.60
Formula C₂₅H₂₄F₃N₃O₆S₂
SMILES

O=S(C1=CC=C([N+]([O-])=O)C(C(F)(F)F)=C1)(NC2=CC=C(/C=C(SC(N3CC4(C)CCCCC4)=O)/C3=O)C=C2)=O
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

Please store the product under the recommended conditions in the Certificate of Analysis.

Purity & Documentation

Select Batch:

Purity: 99.98%

Assay: 99.98%

ee.: 99.98%

Data Sheet (301 KB)
COA (2659 KB)
Elemental Analysis Report (2659 KB)
HNMR (2659 KB)
CNMR (2659 KB)
RP-HPLC (2659 KB)
NP-HPLC (2659 KB)
MS (2659 KB)
LCMS (2659 KB)
IR (2659 KB)
GC (2659 KB)
OR (2659 KB)
SDS-PAGE (2659 KB)
SEC-HPLC (2659 KB)
Other Report (2659 KB)
Handling Instructions (2659 KB)

References

[1]. Chou CC, et al. AMPK reverses the mesenchymal phenotype of cancer cells by targeting the Akt-MDM2-Foxo3a signaling axis. Cancer Res. 2014 Sep 1;74(17):4783-95.

[2]. Trikha P, et al. Targeting myeloid-derived suppressor cells using a novel adenosine monophosphate-activated protein kinase (AMPK) activator. Oncoimmunology. 2016 Jul 25;5(9):e1214787.

[3]. Plews RL, et al. A novel dual AMPK activator/mTOR inhibitor inhibits thyroid cancer cell growth. The Journal of clinical endocrinology and metabolism. 2015 May;100(5):E748-56. [Content Brief]

[4]. Burton MD, et al. Pharmacological activation of AMPK inhibits incision-evoked mechanical hypersensitivity and the development of hyperalgesic priming in mice. Neuroscience. 2017 Sep 17;359:119-129.

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How should lyophilized recombinant proteins be reconstituted and stored?

1. Before opening the cap, centrifuge the vial at 13000 rpm for 20-30 seconds. This step will ensure that any lyophilized powder that may have adhered to the cap or walls is collected at the bottom of the vial, minimizing the risk of product loss.

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OSU-53

All AMPK Isoforms

All mTOR Isoforms

All Akt Isoforms

All IKK Isoforms

All FOXO Isoforms

All ERK Isoforms

Biological Activity

Chemical Information

Purity & Documentation

References

Calculators

Help & FAQs

OSU-53 Related Classifications

HY-125535 Related Classifications

Bioz