Cell lines

H4 human glioma cells stably overexpressing human wild-type APP695

Preparation method

The solubility of this compound in DMSO is >10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37 ℃ for 10 minutes and/or shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.

Reaction Conditions

24 h; 10 μM

Applications

Using H4 cells stably overexpressing human wild-type APP, LY450139 reduced the secretion of Aβ42, Aβ40, and Aβ38 in 96-well-cultured media and increased β-CTF in cell lysates as expected, although this increase was unexpectedly attenuated at high concentrations. The biphasic β-CTF response to LY450139 was also found in Western blot analysis of six-well-cultured cells using two anti-bodies, 82E1 and 6E10, which recognize the N-terminal region of Aβ.

Animal models

Female Tg2576 mice

Dosage form

3 mg/kg, oral taken.

Applications

To determine the sites of β-CTF accumulation in response to LY450139, immunofluorescence of anti-human Aβ/β-CTF N-terminal-specific antibody (82E1) was applied to hippocampal slices from Tg2576 mice. Three-month-old Tg2576 mice were administered 3 mg/kg LY450139 for 8 d, with the brain fixed 3 h after the last administration. When immunofluorescence intensities in these regions were normalized with that in the CA3 stratum pyramidale of the same slice, LY450139-treated mice showed significantly higher relative immunoreactivity compared with the vehicle-treated mice.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

Semagacestat (also known as LY450139), [(2S)-2-hydroxy-N-((2S)-1-((1S)-3-methyl-2-oxo-2,3,4,5-tetrahydro-1H-benzo[d]azepin-1ylamino)-1-oxopropan-2-yl)-3-methylbutanamide] is an azepine class γ-secretase, which is currently being investigated as a potential disease-modifying agent for the treatment of Alzheimer’s disease (AD). It reduces the rate of formation of amyloid-β (Aβ), a major component of the neuritic plaque in the brains of AD patients, in human subjects and animal models including mice, beagle dogs and guinea pigs. According to previous studies, semagacestat slows the accumulation of Aβ in the brains of transgenic mice overexpressing mutant human amyloid precursor protein V717F (PDAPP mice) and exhibits a dose-dependent decrease of plasma in AD patients.

Reference

[1].Ping Yi, Chad Hadden, Palaniappan Kulanthaivel, Nathan Calvert, William Annes, Thomas Brown, Robert J. Barbuch, Archana Chaudhary, Mosun A. Ayan-Oshodi, and Barbara J. Ring. Disposition and metabolism of semagacestat, a γ-secretase inhibitor, in humans. Drug Metabolism and Disposition 2010; 38(4): 554-565