6-Iodopravadoline-AM630-DataSheet-生命科學(xué)試劑-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemE6-IodopravadolineCat. No.: HY-15421CAS No.: 164178-33-0Synonyms: AM630分式: CHINO分量: 504.36作靶點: Cannabinoid Receptor作通路: GPCR/G Protein; Neuronal Signaling儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)

2、體外實驗 DMSO : 50 mg/mL (99.14 mM; Need ultrasonic)Mass Solvent1 mg 5 mg 10 mg Concentration制備儲備液1 mM 1.9827 mL 9.9136 mL 19.8271 mL5 mM 0.3965 mL 1.9827 mL 3.9654 mL10 mM 0.1983 mL 0.9914 mL 1.9827 mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度，選擇合適的溶劑配制儲備液，并請注意儲備液的保存式和期限。體內(nèi)實驗請根據(jù)您的實驗動物和給藥式選擇適當(dāng)?shù)娜芙獍?，配制前請先配制澄的儲備液，再依次添加助溶?為保證實驗結(jié)果的可靠

3、性，體內(nèi)實驗的作液，建議您現(xiàn)現(xiàn)配，當(dāng)天使；澄的儲備液可以根據(jù)儲存條件，適當(dāng)保存；以下溶劑前的百分指該溶劑在您配制終溶液中的體積占)：1. 請依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (4.96 mM); Clear solution2. 請依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (4.96 mM); Clear solution3. 請依序添加每種溶劑： 10% DMSO 90% corn oil

4、1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemESolubility: 2.5 mg/mL (4.96 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 6-Iodopravadoline (AM630)種選擇性的 CB2 拮抗劑，Ki 值為 31.2 nM，對其選擇性對 CB1 受體的 165倍。IC50 & Target Ki: 31.2 nM (CB2)體外研究 The AM251 and 6-Iodopravadoline (AM630)-evoked Ca2+ influxes into TG sen

5、sory neurons aereconcentration-dependent, and fitted. The EC50 for AM251 and 6-Iodopravadoline are 7.37 M and 15.6 M,respectively. AM251 and 6-Iodopravadoline activate TRPA1 in TG sensory neurons. 6-Iodopravadoline iscomparable in value in both TRPA1 and TRPV1/TRPA1 expressing CHO cells (2 and 4.6 M

6、, respectively).AM251 and 6-Iodopravadoline activation of TRPA1 is modulated by TRPV1 2. 6-Iodopravadoline (0, 50,100, and 200 nM) is not toxic to RAW264.7 cells. 6-Iodopravadoline (100 nM) substantially inhibitsosteoclastogenesis in cultures with RANKL and Ti particles in a dose-dependent manner 3.

7、 6-Iodopravadoline (1 M) blocks the CP-55,940 dose response with EC50 of 170 nM at human and EC50 of110 nM at rat cannabinoid CB2 receptor 4.體內(nèi)研究 6-Iodopravadoline (AM630) (2, 3 mg/kg, i.p.) significantly reduces the time spent in the light box comparedwith vehicle group. 6-Iodopravadoline (AM630) (

8、1, 2 or 3 mg/kg, i.p., twice a day) produces a significantanxiolytic effect, increasing the time spent in the light box at all of the doses used 1.PROTOCOLKinase Assay 4 HEK cells stabLy expressing human or rat cannabinoid CB2 receptors are grown until a confLuentmonoLayer is formed. The ceLLs are h

9、arvested and homogenized in Tris-EDTA (TE) buffer (50 mM Tris-HCl,pH 7.4, 1 mM MgCl2, and 1 mM EDTA) using a poLytron for 210 s bursts in the presence of proteaseinhibitors, followed by centrifugation at 45,000g for 20 min. The final membrane pellet is re-homogenized instorage buffer (50 mM Tris-HCl

10、, pH 7.4, 1 mM MgCl2, and 1 mM EDTA and 10% sucrose) and frozen at80C until used. Saturation binding reactions are initiated by the addition of membrane preparation (proteinconcentration of 5 g/well for the human CB2and 20 g/well for the rat CB2) into wells of a deep-well pLatecontaining (3HCP-55,94

11、0 (120 Ci/mmol) in assay buffer (50 mM Tris-HCl, pH 7.4, 2.5 mM EDTA, 5 mMMgCl2, and 0.5 mg/mL fatty acid-free bovine aLbumin serum). After incubation at 30C for 90 min, bindingreaction is terminated by the addition of 300 L/well of coLd assay buffer followed by rapid vacuum filtrationthrough a UniF

12、ilter-96 GF/C filter pLates (pre-soaked in 1 mg/mL bovine serum aLbumin for 2 h). The boundactivity is counted in a TopCount using Microscint-20. In some experiments with endogenous ligands,experiments are performed in the presence of AEBSF (50 M). Saturation experiments are conducted with12 concent

13、rations of 3HCP-55,940 ranging from 0.01 to 8 nM. Competition experiments are conducted with0.5 nM 3HCP-55,940 and five concentrations (1 nM to 10 M) of displacing Ligands. Nonspecific binding isdefined by the addition of 10 M unLabeLed CP-55,940 in both saturation and competition bindingassays. Kd

14、vaLues from saturation binding assays and Ki vaLues from the competition binding assays are2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEdetermined with one site binding or one site competition curve fitting equations using Prism software.MCE has not independently confirmed the accuracy of these

15、methods. They are for reference only.Cell Assay 3 The effect of AM630 and Ti particles on RAW cell viability is examined using the 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazoliumbromide (MTT) assay. RAW 264.7 (1103 cells/well) is cultured in the presence ofAM630 (0, 50, 100, and 200 nM) for 24, 4

16、8, and 72 h, with or without Ti particles, and then incubated withMTT (0.5 mg/mL) at 37C for 2 h. The culture medium is then replaced with equal volumes of DMSO todissolve formazan crystals, followed by shaking at room temperature for 10 min. Absorbance at 490 nm ismeasured using a microplate reader

17、.MCE has not independently confirmed the accuracy of these methods. They are for reference only. EBioMedicine. 2020 Mar 3;53:102696. EBioMedicine. 2020 Mar;53:102696. Biomed Pharmacother. 2019 Feb;110:145-154. Chem Biol Interact. 2018 Oct 19;297:16-24. Eur J Pain. 2017 May;21(5):804-814.See more cus

18、tomer validations on HYPERLINK / www.MedChemEREFERENCES1. Garcia-Gutierrez, Maria S., et al. Chronic blockade of cannabinoid CB2 receptors induces anxiolytic-like actions associated withalterations in GABAA receptors. British Journal of Pharmacology (2012), 165(4), 951-964.2. Patil, Mayur., et al. Cannabinoid receptor antagonists AM251 and AM630 activate TRPA1 in sensory neurons. Neuropharmacology(2011), 61(4), 778-788.3. Geng, D. C., et al. Inhibition of titanium particle-induced inflammatory osteolysis through inactivation of cannabinoid receptor 2 byAM630. Journal of Biomedical Materi