替洛利生鹽酸鹽作用機(jī)制 - Medchemexpress - MCE中國

1、Product Data SheetPitolisant hydrochlorideCat. No.: HY-12199BCAS No.: 903576-44-3分式: CHClNO分量: 332.31作靶點(diǎn): Histamine Receptor作通路: GPCR/G Protein; Immunology/Inflammation; Neuronal Signaling儲(chǔ)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(shí)驗(yàn) H2O : 100 mg/mL (300.92 mM; Need

2、 ultrasonic)DMSO : 43 mg/mL (129.40 mM)* means soluble, but saturation unknown.SolventMass1 mg 5 mg 10 mgConcentration制備儲(chǔ)備液1 mM 3.0092 mL 15.0462 mL 30.0924 mL5 mM 0.6018 mL 3.0092 mL 6.0185 mL10 mM 0.3009 mL 1.5046 mL 3.0092 mL請(qǐng)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(chǔ)備液；旦配成溶液，請(qǐng)分裝保存，避免反復(fù)凍融造成的產(chǎn)品失效。儲(chǔ)備液的保存式和期限：-80C, 6

3、 months; -20C, 1 month。-80C 儲(chǔ)存時(shí)，請(qǐng)?jiān)?6 個(gè)內(nèi)使，-20C 儲(chǔ)存時(shí)，請(qǐng)?jiān)?1 個(gè)內(nèi)使。體內(nèi)實(shí)驗(yàn)請(qǐng)根據(jù)您的實(shí)驗(yàn)動(dòng)物和給藥式選擇適當(dāng)?shù)娜芙獍?。以下溶解案都?qǐng)先按照 In Vitro 式配制澄清的儲(chǔ)備液，再依次添加助溶劑：為保證實(shí)驗(yàn)結(jié)果的可靠性，澄 的儲(chǔ)備液可以根據(jù)儲(chǔ)存條件，適當(dāng)保存；體內(nèi)實(shí)驗(yàn)的作液，建議您現(xiàn)現(xiàn)配，當(dāng)天使； 以下溶劑前顯的百分 指該溶劑在您配制終溶液中的體積占；如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過加熱和/或超聲的式助溶1. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5

4、mg/mL (7.52 mM); Clear solution此案可獲得 2.5 mg/mL (7.52 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲(chǔ)備液加到 900 L 20% 的 SBE-CD 理鹽溶液中，混合均勻。2. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (7.52 mM); Clear solutionPage 1 of 2 www.MedChemE此案可獲得 2.5 mg/mL (7.52 mM，飽和度未知) 的澄 溶液，此案不適于實(shí)驗(yàn)周 期在半

5、個(gè)以上的實(shí)驗(yàn)。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲(chǔ)備液加到 900 L 油中，混合均勻。BIOLOGICAL ACTIVITY物活性 Pitolisant鹽酸鹽(BF2.649;Ciproxidine)新型的重 組H3受體選擇性反向激動(dòng)劑，Ki為0.16 nM。IC & Target Ki: 0.16 nM (H3 receptor)1EC50: 1.5 nM (H3 receptor)1體外研究 On the stimulation of guanosine 5-O-(3-35Sthio)triphosphate binding to this r

6、eceptor, Pitolisant (BF2.649) behavesas a competitive antagonist with a Ki value of 0.16 nM and as an inverse agonist with an EC50 value of 1.5 nM and anintrinsic activity 50% higher than that of ciproxifan. Pitolisant displaces 125Iiodoproxyfan binding from mouse braincortical membranes with an IC5

7、0 value of 26.44.5 nM. Taking into account the Kd value of the radioligand (1619pM), the deduced Ki value for Pitolisant is 141 nM. Pitolisant displaces 125Iiodoproxyfan binding from membranesof rat glioma C6 cells stably expressing the human H3 receptor with an IC50 value of 4.20.2 nM. Taking into

8、accountthe Kd value of the radioligand (504 pM), the deduced Ki value for Pitolisant is 2.70.5 nM. Pitolisant progressivelyreverses this response with a Hill coefficient close to unity and an IC50 value of 33068 nM, leading to a Ki value of174 nM. Pitolisant elicits a dose-dependent decrease of the

9、basal-specific 35SGTPS binding to membranes with amaximal effect corresponding to 751% of the basal-specific binding and an EC50 value of 1.50.1 nM1.體內(nèi)研究 The administration of Pitolisantat a single dose of 10 mg/kg 30 min before a single dose of LY170053 (2 mg/kg b.w.)also significantly affects immo

10、bility time in the FST. Subsequent administration of the aforementioned drug sequencein mice statistically significantly increases the duration of immobility in comparison to the time determined in thecontrol group in the FST. It decreased locomotor activity as well. In contrast, the results obtaine

11、d in subchronictreatment after fifteen administrations of both drugs (Pitolisant 10 mg/kg b.w., and after 30 min LY170053 2 mg/kgb.w., and again after 4 h LY170053 2 mg/kg b.w.) show that the administration of Pitolisant followed by that ofLY170053 equalized the locomotor activity in mice; in compar

12、ison to the level of motility in the control group, towhich only Pitolisant is administered. More importantly, this combination of drugs significantly reduces immobilitytime to the level obtained in the control group in the forced swim test in mice one-way ANOVA; F (3,20)=4.226,P=0.01812. Rats given

13、 Pitolisant (10 mg/kg) during the conditioning phase stayed 50294 s on the pairedtexture, a value not statistically different from that of controls, indicating that Pitolisant did not support placepreference3.PROTOCOLKinase Assay 1 35SGTPS binding assays are performed. CHO-K1 cells stably expressing

14、 the human H3 receptor (400 fmol/mgprotein) are homogenized in ice-cold buffer (50 mM Tris/HCl, pH 7.4). Homogenates are centrifuged twice (20,000gfor 10 min at 4C), and the final pellet is resuspended in 50 volumes of buffer. Membranes (550 g of protein) arepretreated with adenosine deaminase (1 U/

15、mL) and incubated for 60 min at 25C with 0.1 nM 35SGTPS and thedrugs to be tested in a final volume of 1 mL of assay buffer (50 mM Tris/HCl, 50 mM NaCl, 5 mM MgCl2, 10 M GDP,and 0.02% bovine serum albumin, pH 7.4). The nonspecific binding is determined using 10 M nonradioactive GTPS.Incubations are

16、stopped by rapid filtration under vacuum through GF/B glass fiber filters. After washing with ice-coldwater, the radioactivity trapped on filters is counted by liquid scintillation spectrometry. A similar assay is used toassess competitive antagonism. In brief, membranes (10 g of protein) of HEK-293

17、 cells stably expressing the humanH3 receptor (600 fmol/mg protein) are preincubated in presence of Pitolisant in the buffer (50 mM Tris/HCl, pH 7.4,10 mM MgCl2, 100 mM NaCl, and 10 M GDP) in a 96-well microplate under gentle agitation at room temperaturePage 2 of 3 www.MedChemE(19-20C) for 30 min b

18、efore the addition of 0.1 nM 35SGTPS (final volume 200 L). The nonspecific binding isdetermined using a 10 M concentration of nonradioactive GTPS. After 30 min, incubations performed in triplicateare stopped by rapid filtration under vacuum on a Multiscreen MAFCOB50 microplate. Radioactivity trapped

19、 on filtersis counted by liquid scintillation spectrometry1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice2Administration 23 Adult female Albino Swiss mice weighing 20-22 g are used in the study. LY170053 or Pitolisant are suspended in 1 %T

20、ween 80. The compounds or vehicle are administered intraperitoneally (i.p.) 30 min prior to the acute experiment. Inthe Pitolisant+LY170053 group, Pitolisant is administered 15 min before LY170053. Subchronic treatment is done atabout 9:00 am (0.2 mL Tween to control group, Pitolisant-10 mg/kg b.w.

21、to Pitolisant group, LY170053-2 mg/kg b.w.to LY170053 group, Pitolisant-10 mg/kg b.w. and LY170053 after 15 min-2 mg/kg b.w. to Pitolisant+LY170053 group)and at about 1:00 pm (LY170053 group and Pitolisant+LY170053 group).Rats3Male Wistar rats (220-300 g) receive vehicle (methylcellulose 1%, p.o.),

22、and Pitolisant (10 mg/kg, p.o.). Ninety minuteslater, they are killed by decapitation and nucleus accumbens are dissected out, weighed, frozen in liquid nitrogen andstored at -80C. Tissues are homogenized in 1 mL of a 0.4 N perchloric acid/2.7 mM EDTA solution. Aftercentrifugation (8000 rpm, 20 min,

23、 4C), supernatants are analysed by HPLC coupled to electrochemical detection.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻(xiàn) Brain Res. 2020 May 5;1740:146873.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Ligneau X, et al. BF2.649 1-3-3-(4-Chlorophenyl)propoxypro