EN6 Chemical Properties

Boiling point:

484.1±45.0 °C(Predicted)

Density 

1.32±0.1 g/cm3(Predicted)

storage temp. 

2-8°C

solubility 

DMF: 30 mg/ml; DMF:PBS (pH 7.2) (1:10): 0.09 mg/ml; DMSO: 15 mg/ml

form 

A crystalline solid

pka

11.27±0.70(Predicted)

color 

White to off-white

EN6 Usage And Synthesis

Uses

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[1].

Biological Activity

EN6 is an autophagy activator th at simultaneously upregulates lysosomal vacuolar H(+)-ATPase (v-ATPase) activity and induces its decoupling from the Rag GTPases by covalently targeting v-ATPase subunit ATP6V1A Cys277. EN6 blocks mTORC1 lysosomal recruitment & activation (1-4 hr 25 μM EN6 pretreatment prior to 10-min amino acids (AA) stimulation of AA-starived HEK293) and clears cellular TDP-43 aggregates (25 μM; U2OS with inducible TDP-43). When administered in mice in vivo (50 mg/kg ip.), EN6 inhibits mTORC1 signaling and activates autophagy in both skeletal muscle and heart tissue. Unlike bafilomycin A1 (BafA1), EN6 activates, but not inhibits v-ATPase catalytic activity.

in vivo

storage

Store at -20°C

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. DOI:10.1038/s41589-019-0308-4