WWL70 _ABHD6 抑制剂 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEWWL70Cat. No.: HY-100337CAS No.: 947669-91-2分式: CHNO分量: 437.49作靶点: MAGL作通路: Metabolic Enzyme/Protease储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性数据体外实验 DMSO : 17.33 mg/mL (39.61 mM; Need ultrasonic)H

2、2O : 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (5.71 mM); Suspended solution; Need ultrasonicBIOLOGICAL ACTIVITY1/3 Master of Small Molecules 您边的抑制剂师www.MedChemE物活性 WWL70个选择性的 / 解酶结构域 6 (ABHD6) 抑制剂，其 IC50 值为 70 nM。IC50 & Target IC50: 70 nM (ABHD6) 1体外研究 At 1 h after WWL70 (10 M) treatme

3、nt, 2-Arachidonoylglycerol (2-AG) is increased by 20% compare tountreated cells. At either 1 or 10 M, WWL70 completely blocks the lipopolysaccharide (LPS)-inducedincrease of PGE2. The enhanced mRNA expression of mPGES-1 and mPGES-2 by LPS is also reduced byWWL70. The IC50 of WWL70 to inhibit the PGE

4、2 biosynthesis is about 100 nM 2.体内研究 Although post-treatment with WWL70 at 5mg/kg does not have any effect, treatment with WWL70 at 10mg/kgimproves the performance significantly. WWL70 treatment improves motor coordination of traumatic braininjury (TBI) mice in a concentration dependent manner. The

5、 latency to fall in animals treated with WWL70 at 5mg/kg increases from 74.924.8 to 99.575.21 on day 3 (p 3.PROTOCOLKinase Assay 2 One hundred micrograms per milliliter of BV2 microsomes are pre-incubated with WWL70 for 5 min at 23C,then mixed with 10 M of Arachidonic acid (AA) for 1 min at 23C. 500

6、 g/mL brain microsomes areincubated with 10 M of AA for 2 min at 23C. The reaction is stopped by mixing with stannous acid (5mg/mL in 0.1 N HCl) to deactivate the enzyme and convert intermediate PGH2 to PGF2, followed by themeasurement of PGE2 concentration by Enzyme-linked immunoassay (EIA). The ac

7、tivity is determined aftersubtraction with the amount of PGE2 in the microsome fraction incubated without substrate 2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 2 BV2 cells (90% confluence) in 10-cm dishes are treated with WWL70 (10 M) f

8、or 1 h. After rinsing with PBSonce, the cells are collected by centrifugation at 5000 g for 2 min. The pellet is suspended with 0.1 mL of0.02% trifluoroacetic acid (TFA) and 1 nmol of 2-AG-d8 by pipetting and dispersed in 4 mL of acetonitrile in asilanized glass tube to precipitate the debris overni

9、ght at -20C. The supernatant after centrifuged at 5000 gfor 5 min is transferred to a new glass tube and evaporated under a nitrogen gas stream in a mild hot waterbath (approximately 35C). 2-AG is resuspended with 0.1 mL of acetonitrile and stored at -80C until massanalysis 2.MCE has not independent

10、ly confirmed the accuracy of these methods. They are for reference only.Animal Seven-week-old, male C57BL/6 mice weighing 25 to 30g are used in this study. Animals are maintainedAdministration 3 under a controlled environment with a temperature of 232C, a 12h light/dark cycle, and access to food and

11、water ad libitum. WWL70 (5mg/kg or 10mg/kg) in physiologic saline or an equal volume of 1% DMSO insaline (10mL/kg) is injected intraperitoneally, and then once a day for 3, 7, or 21 days depending on theexperimental design. During the 21-day treatment regimen, animals are subjected to a battery of b

12、ehavioraltests at different time points. Two hours after the last injection on day 21 post-injury, animals are sacrificedand brain tissues are collected for histological analysis 3.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES2/3 Master of

13、Small Molecules 您边的抑制剂师www.MedChemE1. Li W, et al. A functional proteomic strategy to discover inhibitors for uncharacterized hydrolases. J Am Chem Soc. 2007 Aug8;129(31):9594-5.2. Tanaka M, et al. WWL70 attenuates PGE2 production derived from 2-arachidonoylglycerol in microglia by ABHD6-independent

14、mechanism. J Neuroinflammation. 2017 Jan 10;14(1):7.3. Tchantchou F, et al. Selective inhibition of alpha/beta-hydrolase domain 6 attenuates neurodegeneration, alleviates blood brain barrierbreakdown, and improves functional recovery in a mouse model of traumatic brain injury. J Neurotrauma. 2013 Apr 1;30(7