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Chemical Structure| 1808714-73-9 Chemical Structure| 1808714-73-9

Structure of EN6
CAS No.: 1808714-73-9

Chemical Structure| 1808714-73-9

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EN6 is a small-molecule activator of autophagy that covalently targets cysteine 277 in the ATP6V1A subunit of the lysosomal v-ATPase, which activates mechanistic target of rapamycin complex 1 (mTORC1) via the Rag guanosine triphosphatases. EN6-mediated ATP6V1A modification decouples the v-ATPase from the Rags, leading to inhibition of mTORC1 signaling, increased lysosomal acidification and activation of autophagy.

4.5 *For Research Use Only! Not for Human Use. We Do Not Sell to Patients.

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Product Details of EN6

CAS No. :1808714-73-9
Formula : C19H14F2N4O2
M.W : 368.34
SMILES Code : O=C(C1=CN(C2=CC=CC=C2F)N=C1)NC3=CC=C(F)C(NC(C=C)=O)=C3
English Name :N-[4-Fluoro-3-(prop-2-enamido)phenyl]-1-(2-fluorophenyl)-1H-pyrazole-4-carboxamide
MDL No. :MFCD31665468
InChI Key :SUSXQEYPNDORDQ-UHFFFAOYSA-N
Pubchem ID :99640033

Safety of EN6

Isoform Comparison

Biological Activity

In Vitro:

Cell Line
Concentration Treated Time Description References
U2OS cells 25 μM 7 hours EN6 treatment reduced TDP-43 aggregates by 75%, and this clearance was lysosome- and v-ATPase-dependent. Nat Chem Biol. 2019 Aug;15(8):776-785
HEK293A cells 25 μM 4 hours EN6 treatment significantly increased the formation of LC3 puncta, similar to Torin1 treatment. Nat Chem Biol. 2019 Aug;15(8):776-785
HEK293A cells 50 μM 24 hours EN6 treatment increased LC3B degradation to a comparable degree to direct mTORC1 inhibitors, rapamycin and Torin1. Nat Chem Biol. 2019 Aug;15(8):776-785
Mycobacterium tuberculosis H37Rv 100 µg/mL 4 hours Evaluate the inhibitory effect of DCS on M. tuberculosis, results showed DCS inhibits two enzymes: alanine racemase (Alr) and D-Ala:D-Ala ligase (Ddl) Nat Commun. 2019 Sep 13;10(1):4177

In Vivo:

Species
Animal Model
Administration Dosage Frequency Description References
C57BL/6 mice Intraperitoneal (i.p.) 50 mg/kg Single dose, analyzed after 4 h EN6 treatment significantly inhibited mTORC1 signaling in skeletal muscle and heart and activated autophagy, as shown by increased LC3BII levels and reduced p62 levels. Nat Chem Biol. 2019 Aug;15(8):776-785

Protocol

Bio Calculators
Preparing Stock Solutions 1mg 5mg 10mg

1 mM

5 mM

10 mM

2.71mL

0.54mL

0.27mL

13.57mL

2.71mL

1.36mL

27.15mL

5.43mL

2.71mL

Dissolving Methods
The prepared working fluid is recommended to be prepared now and used up as soon as possible in a short period of time. The percentage shown in front of the following solvent refers to the volume ratio of the solvent in the final solution; If precipitation or precipitation occurs in the preparation process, it can be assisted by heating and/or ultrasound.
Protocol 1

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