丙磺舒作用机制 - Medchemexpress - MCE中国

1、Product Data SheetProbenecidCat. No.: HY-B0545CAS No.: 57-66-9分式: CHNOS分量: 285.36作靶点: TRP Channel; Bacterial; HIV作通路: Membrane Transporter/Ion Channel; Neuronal Signaling; Anti-infection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 : 100 mg/mL (350.43 mM)H2O :

2、0.1 mg/mL (insoluble)* means soluble, but saturation unknown.SolventMass1 mg 5 mg 10 mgConcentration制备储备液1 mM 3.5043 mL 17.5217 mL 35.0435 mL5 mM 0.7009 mL 3.5043 mL 7.0087 mL10 mM 0.3504 mL 1.7522 mL 3.5043 mL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；旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存式和期限：-80C, 6 months; -20C, 1 m

3、onth。-80C 储存时，请在 6 个内使，-20C 储存时，请在 1 个内使。体内实验请根据您的实验动物和给药式选择适当的溶解案。以下溶解案都请先按照 In Vitro 式配制澄清的储备液，再依次添加助溶剂：为保证实验结果的可靠性，澄 的储备液可以根据储存条件，适当保存；体内实验的作液，建议您现现配，当天使； 以下溶剂前显的百分 指该溶剂在您配制终溶液中的体积占；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的式助溶1. 请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (8.76

4、 mM); Suspended solution; Need ultrasonic此案可获得 2.5 mg/mL (8.76 mM) 的均匀悬浊液，悬浊液可于服和腹腔注射。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 400 L PEG300 中，混合均匀；向上述体系中加50 L Tween-80，混合均匀；然后继续加 450 L 理盐定容 1 mL。2. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (8.76 mM); Clear solutionPage 1 of 2 www.Med

5、ChemE此案可获得 2.5 mg/mL (8.76 mM，饱和度未知) 的澄 溶液，此案不适于实验周 期在半个以上的实验。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 900 L 油中，混合均匀。BIOLOGICAL ACTIVITY物活性 Probenecid。种有效的选择性瞬时受体电位草酸受体通道 2 (TRPV2) 激动剂。Probenecid 还抑制 pannexin 1 通道IC & Target TRPV21体外研究 Probenecid efficiently inhibits ATP-dependent active vesicul

6、ar N-ethylmaleimide glutathione (NEM-GS) uptake byboth MRP1 and MRP2. A significant inhibition of the MRP1-ATPase is observed at higher organic anionconcentrations. In contrast, the ATPase activity of MRP2 is strongly stimulated by both Probenecid (approximate KACT=250 M), sulfinpyrazone (KACT=300 M

7、), and indomethacin (KACT=150 M), and ATPase activation is even strongerthan in the case of NEM-GS. The organic anion activation of the MRP2-ATPase followed bell-shaped curves, withmaximum values obtained at about 2 mM for Probenecid, 800 M for sulfinpyrazone, and 400 M for indomethacin2. Probenecid

8、 is an inhibitor of the hTAS2R16, hTAS2R38, and hTAS2R43 bitter taste receptors. Probenecid acts on asubset of TAS2Rs and inhibits through a novel, allosteric mechanism of action. Probenecid is also commonly used toenhance cellular signals in GPCR calcium mobilization assays. Probenecid specifically

9、 inhibits the cellular responsemediated by the bitter taste receptor hTAS2R16 and provide molecular and pharmacological evidence for directinteraction with this GPCR using a non-competitive (allosteric) mechanism3.体内研究 Administration of Probenecid to WT mice results in increased contractility as mea

10、sured via ejection fraction (EF)relative to EF in control mice given saline. The increased contractility is noted within 5 minutes of the bolus injection with all doses at or above 75 mg/kg (peak change of 5.263.35, 8.402.80, 7.322.52 for 75mg/kg, 100mg/kg and200mg/kg, respectively). The measured ch

11、ange in contractility as measured at 5 minute intervals (for 30 minutes total)revealed a dose dependent increase in contractility with an estimated EC50 of 49.33 mg/kg. The EF remained at anelevated state for at least 1 hour on subjects (n=5, dose of 200 mg/kg IV) that are evaluated for a longer per

12、iod oftime (average increase in EF over baseline of 8.92.57)1.PROTOCOLCell Assay 1 HEK-293T cells are transfected with hTAS2R expression constructs using Lipofectamine 2000 in poly-lysine coated,black 384-well plates with clear bottoms and incubated for 22 hours at 37C. Growth media is removed and c

13、ells arewashed twice with HBSS containing 20 mM HEPES, then loaded with a calcium indicator dye in HBSS containing 20mM HEPES (Calcium 4 Assay kit) with or without 1 mM Probenecid. Cells are incubated at 37C for 1 hour in thepresence of both dye and Probenecid, then moved to a Flexstation II-384 set

14、 for 32C. After a 15-minute temperatureequilibration (without washout), indicated compounds are injected (at t=25 seconds) and fluorescence is measuredfor 100 to 180 seconds, reading every 3 seconds. Data sets are analyzed and represented as % over baseline signalusing Prism 5.0 software. For Schild

15、 plots, replicates of raw calcium flux values are expressed as % over baselinesignal. The mean value at 36 seconds (corresponding to the maximum flux signal) for each concentration of TAS2Rligand in the presence of the indicated concentration of Probenecid is plotted against the log of ligandconcent

16、ration. Data points are fit using non-linear regression in GraphPad Prism1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice1 In order to obtain a dose response curve, male C57 WT (n=39) mice 12-16 weeks of age are anesthetized withAdministrat

17、ion 1 isoflurane while intravenous jugular access (IV) is obtained under a microscope. Subsequently, an echocardiographicPage 2 of 3 www.MedChemEstudy with both M-mode and B-mode is obtained in parasternal long axis (PSLAX) as described below. Either salineor different doses of Probenecid (increasin

18、g from 2 to 200mg/kg) are injected (bolus IV) for the initial contractilitystudies in WT mice.MCE has not independently confirmed the accuracy of these methods. They are for reference only.户使本产品发表的科研献 Cell Mol Immunol. 2019 Mar 25. Int J Mol Sci. 2019 Mar 5;20(5). pii: E1125. J Agric Food Chem. 2016

19、 Oct 26;64(42):7899-7910. Lasers Surg Med. 2017 Sep;49(7):719-726. Regul Toxicol Phar. 2019 Aug 23:104449.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Koch SE, et al. Probenecid: novel use as a non-injurious positive inotrope acting via cardiac TRPV2 stimulation. J Mol Cell Cardiol. 2012 Jul;53(1):134-44.2. Bakos E, et al. Int