SB-408124

• 产品号: S1545
• CAS号: 288150-92-5
• 分子式: C19H18F2N4O
• 分子量: 356.3692264
• 保 存: -20°C

产品别名

• 标题: SB-408124
• IUPAC标准名: 3-(6,8-difluoro-2-methylquinolin-4-yl)-1-[4-(dimethylamino)phenyl]urea
• IUPAC传统名: 3-(6,8-difluoro-2-methylquinolin-4-yl)-1-[4-(dimethylamino)phenyl]urea
• 别名: SB408124

产品登记号

• CAS号: 288150-92-5

产品性质

• 作用靶点: OX1
• 成盐信息: Free Base
• 保存条件: -20°C

产品详细信息

详细说明 (English)

Biological Activity

• Description: SB-408124 (Tocris-1963) is a non-peptide antagonist for OX1 with K_i of 57 nM and 27 nM in both whole cell and membrane, respectively.
• Targets:

  OX1 (whole cell)

  ;

  OX1 (membrane)

• IC50:

  57 nM (K_i)

  ;

  27 nM (K_i) ^[1]

• In Vitro: SB-408124 binds hypocretin type 1 receptor (HcrtR1) with pK_i values of 7.57. Calcium mobilization studies shows that SB-408124 is a functional antagonist of the OX1 receptor with a affinity of approximately 50-fold selectivity over the OX2 receptor. ^[1] A recent study indicates that pretreatment of primary cultures of rat astrocytes with SB-401824 before Orexin A administration significantly reduced the stimulatory action of Orexin A on both basal and forskolin-acivated cAMP production. ^[2]
• In Vivo: SB-408124 (30 μg/10 μL, administered intracerebroventricularly) decreases Orexin-A induced water intake in Wistar rats. Intracerebroventricularly administered Orexin-A (30 μg/10 μL) blocks the vasopressin (VP) level increase induced by either histamine or 2.5% NaCl administration, and this blocking effect is moderated by pretreatment with SB-408124. ^[3] Intracerebroventricular pretreatment with SB-408124 (50 mM, 5 μL/h) prevents Bicuculline (BIC)-induced increases in endogenous glucose production (EGP). ^[4]

Protocol

• Protocol: Kinase Assay ^[1]
• Whole cell binding assays: After overnight culture in 96-well Packard Cultur plates, the medium is discarded and cells are incubated in buffer containing 150 mM NaCl, 20 mM HEPES and 0.5% bovine serum albumin (pH 7.4) for 60 minutes at 25 °C. Saturation studies are carried out by incubating cells with a range of concentrations of [^3H]SB-408124 (0.2–24 nM); the total assay volume is 250 μL. Protein content is assayed by lysing cells with 0.1M NaOH and using the Bradford method with bovine serum albumin (BSA) as a standard. Association kinetic studies are performed by measuring the specific binding of [^3H]SB-408124 (3 nM) at 1–60 minutes after addition of [^3H]SB-408124. All assays are terminated by washing the cells three times with 250μL ice-cold phosphate-buffered saline. A volume of 100 μL of Microscint 40 is added to each well and the plate is left at room temperature for 2 hours. Cell-associated radioactivity is then measured using a Packard Topcount, with a count time of 2 minute/well.
• Membrane-based SPA binding assays: CHO-K1_OX1 cell membranes (75 μg/mL) are precoupled by shaking with wheatgerm-agglutinin polyvinyltoluene (WGA-PVT) scintillation proximity assay (SPA) beads (5 mg/mL) in buffer containing 25 mM HEPES, 2.5 mM MgCl₂, 0.5 mM EDTA and 0.025% bacitracin (pH 7.4) at 4 °C for 1 hour. The bead-membrane suspension is centrifuged at 300× g and resuspended in the same volume of room temperature assay buffer. A volume of 100 μL of bead-membrane suspension is incubated with [^3H]SB-674042 (5 nM) in a total assay volume of 200 μL in a 96-well Packard Optiplate to give a final protein concentration of 7.5 μg/well. Nonspecific binding is measured as that remaining in the presence of 3 μM SB-408124. Assay plates are shaken for 10 min and then incubated at room temperature for 4 hours before being counted on a Packard TopCount scintillation counter (count time 2 min/well). Saturation studies are carried out by incubating bead-membranes (equivalent to 7.5 μg protein/well and 2.5 mg beads/mL) with a range of concentrations of [^3H]SB-674042 (0.1–20 nM). Protein content is assayed using the Bradford method using bovine serum albumin as a standard. Association kinetic studies are performed by measuring specific binding of [^3H]SB-674042 (5 nM) at 1–30 min after addition of bead-membranes (equivalent to 7.5 μg protein/well and 2.5 mg beads/mL). For dissociation studies, bead-membranes are first incubated with [^3H]SB-674042 (5 nM) for 30 min. Specific binding is then measured at 2–120 min after the addition of 3 μM SB-408124. Competition studies are performed by incubating bead-membranes (equivalent to 7.5 μg protein/well and 2.5 mg beads/mL) with [^3H]SB-674042 (5 nM) and a range of concentrations of the test compound.
• Protocol: Animal Study ^[3]
• Animal Models: Male Wistar rats
• Formulation: SB-408124 is dissolved in saline.
• Doses: 30 μg/10 μL
• Administration: Intracerebroventricularly (i.c.v.) injected into the lateral ventricle of the rat.

References

• References: [1] Langmead CJ, et al, Br J Pharmacol, 2004, 141(2) , 340-346.
• References: [2] Woldan-Tambor A, et al, Pharmacol Rep, 2011, 63(3), 717-723.
• References: [3] Kis Gk, et al, Pflugers Arch, 2012, 463(4), 531-536
• References: [4] Yi CX, et al, Diabetes, 2009, 58(9), 1998-2005.

参考文献

• Langmead CJ, et al, Br J Pharmacol, 2004, 141(2) , 340-346.
• Woldan-Tambor A, et al, Pharmacol Rep, 2011, 63(3), 717-723.
• Kis Gk, et al, Pflugers Arch, 2012, 463(4), 531-536
• Yi CX, et al, Diabetes, 2009, 58(9), 1998-2005.