Cat. No.: HY-128892

EN6

5 Cited Publications

Customer Review

Based on 5 publication(s) in Google Scholar

EN6 is a small-molecule in vivo autophagy activator that covalently targets cysteine 277 in the ATP6V1A subunit of the lysosomal v-ATPase. EN6-mediated modification of ATP6V1A uncouples v-ATPase from Rag, leading to inhibition of mTORC1 signalling, increased lysosomal acidification, and activation of autophagy. EN6 also scavenges TDP-43 aggregates (causative agents of frontotemporal dementia) in a lysosome-dependent manner.

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

Purity: 99.42%
CAS No.: 1808714-73-9
Formula: C₁₉H₁₄F₂N₄O₂
Molecular Weight:368.34

Storage:

Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 2 years , -20°C, 1 year

EN6 Chemical Structure

CAS No.: 1808714-73-9

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Top Publications Citing

Use of MCE Products

Fluorescent Dye

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

Biological Activity
- Description
- In Vitro
- In Vivo
Chemical Information
Publications (5)
Solvent & Solubility
- In Vitro
- In Vivo
Purity & Documentation
References
Help & FAQs
Classifications
Bioz

Biological Activity

Description

In Vitro

EN6 (50 μM; 1, 4, 8 h) increases the levels of LC3BII, and triggers formation of LC3 puncta in a time- and dose-dependent manner, in HEK293A cells^[1].
? EN6 leads to significant increases in the number autophagosomes and autolysosomes in HEK293A cells^[1].
? EN6 (25 μM; 1 h) blocks mTORC1 lysosomal localization and activation in HEK293A cells^[1].
? EN6 (50 μM; 4 h) activates the v-ATPase and lysosome acidification in HEK293A cells^[1].
? EN6 (25 μM; 7 h) promotes autophagic clearance of protein aggregates in IPTG-inducible GFP-TDP43 U2OS osteosarcoma cell line model^[1].

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

Western Blot Analysis^[1]

Cell Line:

HEK293A cells
Concentration:

50 μM
Incubation Time:

1, 4, 8 h
Result:

Time- and dose-dependently triggered formation of LC3 puncta and increased the levels of LC3BII.

Western Blot Analysis^[1]

Cell Line:

HEK293A cells
Concentration:

25 μM
Incubation Time:

1 h
Result:

Led to complete inactivation of mTORC1 signaling, as shown by reduced levels of phosphorylated canonical substrates, S6 kinase 1 (S6K1), 4EBP1, and ULK1.

Immunofluorescence^[1]

Cell Line:

HEK293A cells
Concentration:

50 μM
Incubation Time:

4 h
Result:

Led to significantly increased acidification of the lysosome in HEK293A cells, and this heightened acidification was blocked by BafA1.

Immunofluorescence^[1]

Cell Line:

IPTG-inducible GFP-TDP43 U2OS osteosarcoma cell line model
Concentration:

25 μM
Incubation Time:

7 h
Result:

Reduced IPTG-induced TDP43 aggregates by 75 %.

In Vivo

EN6 (50 mg/kg; i.p.; single) inhibits mTORC1 and activates autophagy in vivo^[1].

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

Animal Model:

Six-week-old male C57BL/6 mice^[1].
Dosage:

50 mg/kg
Administration:

Intraperitoneal injection; single
Result:

Significantly inhibited mTORC1 signaling in both skeletal muscle and heart, as demonstrated by reduced phosphorylation of S6, 4EBP1 and ULK1.
Strongly activated autophagy as shown by heightened LC3BII levels and reduced p62 levels.

Chemical Information

CAS No. 1808714-73-9
Appearance Solid
Molecular Weight 368.34
Formula C₁₉H₁₄F₂N₄O₂
Color White to off-white
SMILES

O=C(C1=CN(C2=CC=CC=C2F)N=C1)NC3=CC=C(F)C(NC(C=C)=O)=C3
Shipping

Room temperature in continental US; may vary elsewhere.

Storage

Powder	-20°C	3 years
	4°C	2 years
In solvent	-80°C	2 years
	-20°C	1 year

Publications (5)

All Cell Imaging/StainingWBIFHistological Imaging/Staining

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Adv Sci (Weinh)

Exploring the Link Between Autophagy-Lysosomal Dysfunction and Early Heterotopic Ossification in Tendons. [Abstract]

2024 May 13:e2400790. PMID: 38741381

EN6 purchased from MedChemExpress. Usage Cited in: Adv Sci (Weinh). 2024 May 13:e2400790. [Abstract]

EN6 (50 mg/kg; every other day for 14 d) significantly impaired the level of extracellular matrix calcification of tendon fibroblasts.

EN6 purchased from MedChemExpress. Usage Cited in: Adv Sci (Weinh). 2024 May 13:e2400790. [Abstract]

EN6 (50 mg/kg; every other day for 14 d) reduced extracellular vesicles specific protein CD9 and autolysosomes associated LC3 and LAMP1 in tendon fibroblasts treated with IL‐1β+TNF‐α.

EN6 purchased from MedChemExpress. Usage Cited in: Adv Sci (Weinh). 2024 May 13:e2400790. [Abstract]

EN6 (50 mg/kg; every other day) decreased the green fluorescent signal in the Achilles tendon of C57BL/6J mice, suggesting that autophagosomes were hydrolyzed after binding to lysosomes and autophagic flux was restored. Impaired autophagic flux (yellow arrows) and normal autophagic flux (red arrows).

EN6 purchased from MedChemExpress. Usage Cited in: Adv Sci (Weinh). 2024 May 13:e2400790. [Abstract]

EN6 (50 mg/kg; every other day) significantly decreased the expression level of LC3B and P62 in the Achilles tendons of C57BL/6J mice, suggesting restoration of autophagic flux.

EN6 purchased from MedChemExpress. Usage Cited in: Adv Sci (Weinh). 2024 May 13:e2400790. [Abstract]

EN6 (50 mg/kg; every other day) significantly reduced heterotopic ossification (HO) in tendons of C57BL/6J mice at 1 and 3 weeks after surgery.
Proc Natl Acad Sci U S A

Endocytosis triggers V-ATPase-SYK-mediated priming of cGAS activation and innate immune response. [Abstract]

2022 Oct 25;119(43):e2207280119. PMID: 36252040
Neurochem Res

3BDO Alleviates Seizures and Improves Cognitive Function by Regulating Autophagy in Pentylenetetrazol (PTZ)-Kindled Epileptic Mice Model. [Abstract]

2022 Dec;47(12):3777-3791. PMID: 36243819
Res Sq

2026 Jan 9.
bioRxiv

2025 June 03.

Solvent & Solubility

In Vitro:

DMSO : 5 mg/mL (13.57 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Ethanol : 1.11 mg/mL (3.01 mM; ultrasonic and warming and heat to 60°C)

View the Complete Stock Solution Preparation Table

Preparing Stock Solutions

View the Complete Stock Solution Preparation Table

Concentration Solvent Mass

1 mg

5 mg

10 mg

Concentration

Solvent

Mass

1 mM

2.7149 mL

13.5744 mL

27.1488 mL

5 mM

0.5430 mL

2.7149 mL

5.4298 mL

10 mM

0.2715 mL

1.3574 mL

2.7149 mL

Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.

Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

Molarity Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

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Dilution Calculator

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In Vivo:

Working Solution

For the following dissolution methods, please prepare the working solution directly:

It is recommended to prepare fresh solutions and use them promptly within a short period of time.

The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: 50% PEG300 50% Saline

Solubility: 10 mg/mL (27.15 mM); Suspended solution; Need ultrasonic

Purity & Documentation

Select Batch:

Purity: 99.98%

Assay: 99.98%

ee.: 99.98%

Data Sheet (277 KB)
SDS (396 KB)
- English - EN (396 KB)
- Français - FR (396 KB)
- Deutsch - DE (396 KB)
- Norwegian - NO (396 KB)
- Español - ES (396 KB)
- Swedish - SV (396 KB)
- Italian - IT (396 KB)
- Korean - KR (396 KB)
- Portuguese - PT (396 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]. Chung CY, et al. Covalent targeting of the vacuolar H+-ATPase activates autophagy via mTORC1 inhibition. Nat Chem Biol. 2019 Aug;15(8):776-785. [Content Brief]

Help & FAQs

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.

Complete Stock Solution Preparation Table

Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg
Ethanol / DMSO	1 mM	2.7149 mL	13.5744 mL	27.1488 mL	67.8721 mL
DMSO	5 mM	0.5430 mL	2.7149 mL	5.4298 mL	13.5744 mL
DMSO	10 mM	0.2715 mL	1.3574 mL	2.7149 mL	6.7872 mL

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EN6

Autophagy Compound Library

Anti-Aging Compound Library

Covalent Screening Library

Cysteine Targeted Covalent Library

Cell Death Library

Bioactive Compound Library Max

MCE Bioactive Compound Library

Covalent Screening Library Plus

Bioactive Compound Library

High-Throughput Bioactive Compound Library

Publications Citing Use of MedChemExpress (MCE) EN6

Biological Activity

Chemical Information

Publications (5)

Solvent & Solubility

Purity & Documentation

References

Help & FAQs

Complete Stock Solution Preparation Table

EN6

Autophagy Compound Library

Anti-Aging Compound Library

Covalent Screening Library

Cysteine Targeted Covalent Library

Cell Death Library

Bioactive Compound Library Max

MCE Bioactive Compound Library

Covalent Screening Library Plus

Bioactive Compound Library

High-Throughput Bioactive Compound Library

Publications Citing Use of MedChemExpress (MCE) EN6

Biological Activity

Chemical Information

Publications (5)

Solvent & Solubility

Purity & Documentation

References

Help & FAQs

Complete Stock Solution Preparation Table

EN6 Related Classifications

HY-128892 Related Classifications