Nuciferine-DataSheet-生命科學(xué)試劑-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemENuciferineCat. No.: HY-N0049CAS No.: 475-83-2分式: CHNO分量: 295.38作靶點: 5-HT Receptor; Dopamine 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)體外實驗 DMSO : 11

2、.11 mg/mL (37.61 mM; Need ultrasonic)H2O : 40% PEG300 5% Tween-80 45% salineSolubility: 1.11 mg/mL (3.76 mM); Clear solution2. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 1.11 mg/mL (3.76 mM); Clear solution1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEBIOLOGICAL ACTIVITY物活性 Nuciferine種 5-HT2A，5-

3、HT2C 和 5-HT2B 拮抗劑，IC50 分別為 478 nM，131 nM 和 1 M；也5-HT7的反向激動劑，IC50 為 150 nM。IC50 & Target IC50: 131 nM (5-HT2C receptor), 150 nM (5-HT7 receptor), 478 nM (5-HT2A receptor), 1 M (5-HT2Breceptor) 1EC50: 64 nM (D2 receptor), 2.6 M (D5 receptor), 700 nM (5-HT6 receptor),3.2 M (5-HT1A receptor), 2 M(D4 rec

4、eptor) 1體外研究 Nuciferine is a partial agonist at DD2 receptor with an activity (Emax=67% of dopamine) similar toaripiprazole (Emax=50% of dopamine). In line with its partial agonist activity, Nuciferine inhibited dopamine-induced activation of Gi with a potency similar to clozapine (Nuciferine KB=62

5、nM; Clozapine KB=20 nM) asdetermined via Schild regression analysis 1. The natural product Nuciferine acts as an effective inhibitor ofadult worm motility. Nuciferine is effective at inhibiting both basal and 5-HT evoked motility of adultschistosomes. Nuciferine inhibits Sm.5HTRL and schistosomule w

6、ith 0.240.04 and 0.620.22 M,respectively 2.體內(nèi)研究 In rodent models relevant to antipsychotic drug action, Nuciferine blocks head-twitch responses anddiscriminative stimulus effects of a 5-HT2A agonist, substituted for clozapine discriminative stimulus,enhanced amphetamine induced locomotor activity, i

7、nhibited phencyclidine (PCP)-induced locomotor activity,and rescued PCP-induced disruption of prepulse inhibition without induction of catalepsy. In the presence of1 or 3 mg/kg Nuciferine, cumulative PCP doses produce similar substitution to PCP alone. In the clozapine-trained animals, a dose-depend

8、ent substitution for 1.25 mg/kg clozapine is seen at 10 mg/kg Nuciferine(80.63% drug lever responding), with an ED50 value of 5.42 mg/kg (95% CI 3.09-9.48 mg/kg) while the lowerdoses tested (0.1 mg/kg-3 mg/kg) fails to produce substitution for clozapines discriminative cue. In additionto a high perc

9、entage of responding on the clozapine-appropriate lever, 10 mg/kg Nuciferine also producessignificant rate suppression as compared to vehicle control points (p 1.PROTOCOLKinase Assay 1 For affinity determination, Nuciferine is subjected to primary radioligand binding assays tested at a single 10M co

10、ncentration to displace 50% of the radioligand at a given receptor target. If a more than 50% of theradioligand is displaced, Nuciferine is selected for a secondary binding assay tested at 11 concentrations intriplicate in competition with the radioligand to generate an IC50 and Ki. Binding assays a

11、re performed in 96-well plates with 125 L per well in appropriate binding buffer using radioligand at or near the Kd. Plates areincubated at room temperature in the dark for 90 min. Reactions are stopped by vacuum filtrations onto 0.3%polyethyleneimine soaked 96-well filter mats using a 96-well Filt

12、ermate harvester, followed by at least threewashes of cold wash buffer. Scintillation cocktail is melted onto dried filters and radioactivity is counted usinga Wallac Trilux Microbeta 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 Cells

13、are plated into 48-well plates one day before uptake is performed. Cells are washed with 0.5 mL uptakebuffer (4 mM Tris, 6.25 mM HEPES, 120 mM NaCl, 5 mM KCl, 1.2 mM CaCl2, 1.2 mM MgSO4, 5.6 mM D-2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEglucose, 1.7 mM ascorbic acid, and 1 M pargyline, pH 7.

14、4). Cells are incubated with 225 L uptake bufferwith or without the indicated concentration of Nuciferine for 15 minutes. After incubation, 25 L uptake buffercontaining 3H-DA and DA is added for a final concentration of 20 nM 3H-DA and 1 M DA. Cells areincubated at 37C for 20 minutes or for the time

15、 indicated. Nonspecific uptake is determined in the presenceof 10 M nomifensine. Uptake is terminated by aspirating uptake buffer and washing each well twice with 0.5mL ice-cold uptake buffer. Cells are lysed in 0.1 N NaOH and transferred to vials containing 3 mL scintillationcocktail. Radioactivity

16、 is quantitated using a Beckman LS6500 counter. Data are analyzed in Graph PadPrism 5.0 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice 1Administration 1 Adult male NIH Swiss mice weighing approximately 25 g are used. Mice are injected wit

17、h either Nuciferine (1,3, or 10 mg/kg, i.p.) or vehicle, n=4 mice/condition. Fifteen minutes later, mice are injected with 1 mg/kg DOI(i.p.) and immediately placed in an observation chamber (new cage without bedding). Head-twitches(operationally defined as a rapid rotational jerk of the head that ca

18、n be distinguished from species-appropriate grooming or scratching behaviors) are counted for 20 minutes in 5 minute bins. For the time-course study, mice are pretreated with 3.0 mg/kg Nuciferine (i.p.) at 60, 45, 30, 15, or 0 minutes (co-injection)prior to the 1.0 mg/kg DOI (i.p.) injection, and he

19、ad-twitches are counted as described above. In oneexperiment, mice (n=4 per condition) are pretreated with an injection (s.c.) of 3.0 mg/kg Nuciferine or vehicle15 minutes prior to 1.0 mg/kg DOI injection (i.p.) and head-twitches are counted as described above. Allexperiments are performed by 3 obse

20、rvers, with 2 observers blinded to the experimental conditions which areevenly distributed. Power analyses are performed with the resulting data. The two highest doses ofNuciferine tested (10 and 3 mg/kg), had 0.96 and 0.88 power to detect significance (=0.05). As theseexperiments are performed blinded and in distinct mice, further re