EN6
目录号 : GC35982
An activator of autophagy
Cas No.:1808714-73-9
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >99.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
EN6 is an activator of autophagy.1 It inhibits mammalian target of rapamycin complex 1 (mTORC1) signaling by binding covalently to cysteine 277 of the ATP6V1A catalytic subunit of the lysosomal vacuolar ATPase (v-ATPase) and impairing v-ATPase coupling with Rag GTPases. It is selective for inhibition of mTORC1 over mTORC2 signaling. EN6 increases lysosome acidification and increases cellular clearance of TDP-43 protein aggregates, which are a pathological feature of several neurodegenerative diseases, in a v-ATPase-dependent manner in U2OS osteosarcoma cells. It also inhibits mTORC1 signaling and enhances autophagy in mouse skeletal muscle and heart when administered at a dose of 50 mg/kg.
1.Chung, C.Y.-S., Shin, H.R., Berdan, C.A., et al.Covalent targeting of the vacuolar H+-ATPase activates autophagy via mTORC1 inhibitionNat. Chem. Biol.15776-785(2019)
| Cas No. | 1808714-73-9 | SDF | |
| Canonical SMILES | O=C(C1=CN(C2=CC=CC=C2F)N=C1)NC3=CC=C(F)C(NC(C=C)=O)=C3 | ||
| 分子式 | C19H14F2N4O2 | 分子量 | 368.34 |
| 溶解度 | DMSO: 5 mg/mL (13.57 mM) | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.7149 mL | 13.5744 mL | 27.1488 mL |
| 5 mM | 0.543 mL | 2.7149 mL | 5.4298 mL |
| 10 mM | 0.2715 mL | 1.3574 mL | 2.7149 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Marinobacter goseongensis sp. nov., from seawater
Int J Syst Evol Microbiol 2008 Dec;58(Pt 12):2866-70.PMID:19060073DOI:10.1099/ijs.0.65522-0.
A Gram-negative marine bacterium, designated strain EN6(T), was isolated from seawater of the East Sea of Korea. The organism grew in 1-25 % (w/v) NaCl and at 10-37 degrees C and pH 5.3-9.3, with optimal growth occurring in 4-5 % NaCl and at 25-30 degrees C and pH 7.5. Phylogenetic analysis of the 16S rRNA gene sequence of strain EN6(T) placed this bacterium in the clade Marinobacter within the class Gammaproteobacteria. The 16S rRNA gene sequence similarity between strain EN6(T) and Marinobacter lipolyticus SM19(T), the most closely related species, was 98.4 %, and the level of DNA-DNA relatedness between the two strains was 22 %. On the basis of the phylogenetic analysis and phenotypic and chemotaxonomic data, strain EN6(T) is considered to represent a novel species of the genus Marinobacter. The name Marinobacter goseongensis sp. nov. is proposed, with strain EN6(T) (=KCTC 12515(T)=DSM 19471(T)) as the type strain.
Covalent targeting of the vacuolar H+-ATPase activates autophagy via mTORC1 inhibition
Nat Chem Biol 2019 Aug;15(8):776-785.PMID:31285595DOI:10.1038/s41589-019-0308-4.
Autophagy is a lysosomal degradation pathway that eliminates aggregated proteins and damaged organelles to maintain cellular homeostasis. A major route for activating autophagy involves inhibition of the mTORC1 kinase, but current mTORC1-targeting compounds do not allow complete and selective mTORC1 blockade. Here, we have coupled screening of a covalent ligand library with activity-based protein profiling to discover EN6, a small-molecule in vivo activator of autophagy that covalently targets cysteine 277 in the ATP6V1A subunit of the lysosomal v-ATPase, which activates 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. Consistently, EN6 clears TDP-43 aggregates, a causative agent in frontotemporal dementia, in a lysosome-dependent manner. Our results provide insight into how the v-ATPase regulates mTORC1, and reveal a unique approach for enhancing cellular clearance based on covalent inhibition of lysosomal mTORC1 signaling.
3BDO Alleviates Seizures and Improves Cognitive Function by Regulating Autophagy in Pentylenetetrazol (PTZ)-Kindled Epileptic Mice Model
Neurochem Res 2022 Dec;47(12):3777-3791.PMID:36243819DOI:10.1007/s11064-022-03778-8.
3-benzyl-5-((2-nitrophenoxy) methyl)-dihydrofuran-2(3 H)-one (3BDO) is a mTOR agonist that inhibits autophagy. The main purpose of this study is to investigate the effects of 3BDO on seizure and cognitive function by autophagy regulation in pentylenetetrazol (PTZ)-kindled epileptic mice model. The PTZ-kindled epileptic mice model was used in study. The behavioral changes and electroencephalogram (EEG) of the mice in each group were observed. The cognitive functions were tested by Morris water maze test. The loss of hippocampal neurons was detected by hematoxylin-eosin (HE) staining and immunofluorescence analysis. Immunohistochemistry, western blot and q-PCR were employed to detect the expression of autophagy-related proteins and mTOR in the hippocampus and cortex. Less seizures, increased hippocampal neurons and reduced astrocytes of hippocampus were observed in the 3BDO-treated epileptic mice than in the PTZ-kindled epileptic mice. Morris water maze test results showed that 3BDO significantly improved the cognitive function of the PTZ-kindled epileptic mice. Western blot analyses and q-PCR revealed that 3BDO inhibited the expression of LC3, Beclin-1, Atg5, Atg7 and p-ULK1/ULK1, but increased that of p-mTOR/mTOR, p-P70S6K/P70S6K in the hippocampus and temporal lobe cortex of epileptic mice. Immunohistochemistry and immunofluorescence also showed 3BDO inhibited the LC3 expression and increased the mTOR expression in the hippocampus of epileptic mice. In addition, the autophagy activator EN6 reversed the decrease in the 3BDO-induced autophagy and aggravated the seizures and cognitive dysfunction in the epileptic mice. 3BDO regulates autophagy by activating the mTOR signaling pathway in PTZ-kindled epileptic mice model, thereby alleviating hippocampus neuronal loss and astrocytes proliferation, reducing seizures and effectively improving cognitive function. Therefore, 3BDO may have potential value in the treatment of epilepsy.
GSK-3β inhibition elicits a neuroprotection by restoring lysosomal dysfunction in neurons via facilitation of TFEB nuclear translocation after ischemic stroke
Brain Res 2022 Mar 1;1778:147768.PMID:34968440DOI:10.1016/j.brainres.2021.147768.
Lysosomal dysfunction is an essential pathogenesis of autophagic neuronal injury after ischemic stroke. As a result of cerebral ischemia, transcription factor EB (TFEB) is greatly phosphorylated by prominently activated glycogen synthase kinase-3β (GSK-3β). This increased TFEB phosphorylation decreases its nuclear translocation and subsequently leads to reduced lysosomal biosynthesis, which ultimately results in lysosomal dysfunction. The present study is to investigate whether the lysosomal dysfunction in neurons can be restored to alleviate post-stroke damage by GSK-3β inhibition. The GSK-3β activity was inhibited by pre-treatment with CHIR-99021 (CHIR) for 3 days before middle cerebral artery occlusion (MCAO) surgery in rats. Besides, the lysosomal capacity was altered by pre-administration with Bafilomycin A1 (Baf-A1) and EN6, respectively. Twenty-four hours after MCAO/reperfusion, the penumbral tissues were obtained to detect the GSK-3β, cytoplasmic and nuclear TFEB, and proteins in autophagic/lysosomal pathway by western blot and immunofluorescence, respectively. Meanwhile, the infarct volume, neurological deficits and neuron survival were assessed to evaluate the neurological outcomes elicited by GSK-3β inhibition. The results demonstrated that the neurological injury could be significantly mitigated by GSK-3β inhibition in MCAO + CHIR group, compared with that in MCAO group. Moreover, CHIR-facilitated TFEB nuclear translocation in neurons was coupled with reinforced lysosomal activities and attenuated autophagic substrates. However, GSK-3β inhibition-induced neuroprotection was greatly counteracted by Baf-A1-weakened lysosomal capacity. Conversely, EN6-reinforced lysosomal activities further ameliorated the autophagic/lysosomal signaling, and synergistically alleviated the neurological damage upon GSK-3β inhibition after MCAO/reperfusion. Our data suggests that GSK-3β inhibition-augmented neuroprotection against ischemic stroke is elicited by restoring the lysosomal dysfunction in neurons.
Safety effects of low-cost engineering measures. An observational study in a Portuguese multilane road
Accid Anal Prev 2012 Sep;48:346-52.PMID:22664700DOI:10.1016/j.aap.2012.02.004.
Single carriageway multilane roads are not, in general, a very safe type of road, mainly because of the high number of seriously injured victims in head-on collisions, when compared with dual carriageway multilane roads, with a median barrier. In this paper the results of a study on the effect of the application of several low cost engineering measures, aimed at road infrastructure correction and road safety improvement on a multilane road (EN6), are presented. The study was developed by the National Laboratory of Civil Engineering (LNEC) for the Portuguese Road Administration and involved a comparison of selected aspects of motorized traffic behaviour (traffic volumes and speeds) measured in several sections of EN6, as well as monitoring of road safety developments in the same road. The applied low cost engineering measures allowed a reduction of 10% in the expected annual number of personal injury accidents and a 70% decrease in the expected annual number of head-on collisions; the expected annual frequency of accidents involving killed and seriously injured persons was reduced by 26%.