Small molecule LX2343 ameliorates cognitive deficits in AD model mice by targeting both amyloid β production and clearance


BIBLIOGRAPHIC THERAPEUTIC AGENT ANIMAL MODEL EXPERIMENTAL DESIGN OUTCOMES

Bibliographic

Year of Publication:
2016
Contact PI Name:
Xu Shen
Contact PI Affiliation:
CAS Key Laboratory of Receptor Research and State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai and University of Chinese Academy of Sciences, Beijing, China
Co-Authors:
Xiao-dan Guo, Guang-long Sun, Ting-ting Zhou, Xin Xu, Zhi-yuan Zhu, Vatcharin Rukachaisirikul, Li-hong Hu
Primary Reference (PubMED ID):
Funding Source:
National Natural Science Foundation of China
NSFC-TRF Collaboration Projects
Drug Innovation Project of SIMM
Study Goal and Principal Findings:

Aim: Streptozotocin (STZ) is widely used to induce oxidative damage and to impair glucose metabolism, apoptosis, and tau/Aβ pathology, eventually leading to cognitive deficits in both in vitro and in vivo models of Alzheimer's disease (AD). In this study, we constructed a cell-based platform using STZ to induce stress conditions mimicking the complicated pathologies of AD in vitro, and evaluated the anti-amyloid effects of a small molecule, N-(1,3-benzodioxol-5-yl)-2-[5-chloro-2-methoxy(phenylsulfonyl)anilino]acetamide (LX2343) in the amelioration of cognitive deficits in AD model mice.

Methods: Cell-based assays for screening anti-amyloid compounds were established by assessing Aβ accumulation in HEK293-APPswand CHO-APP cells, and Aβ clearance in primary astrocytes and SH-SY5Y cells after the cells were treated with STZ in the presence of the test compounds. Autophagic flux was observed using confocal laser scanning microscopy. APP/PS1 transgenic mice were administered LX2343 (10 mg·kg-1·d-1, ip) for 100 d. After LX2343 administration, cognitive ability of the mice was evaluated using Morris water maze test, and senile plaques in the brains were detected using Thioflavine S staining. ELISA assay was used to evaluate Aβ and sAPPβ levels, while Western blot analysis was used to measure the signaling proteins in both cell and animal brains.

Results: LX2343 (5-20 μmol/L) dose-dependently decreased Aβ accumulation in HEK293-APPsw and CHO-APP cells, and promoted Aβ clearance in SH-SY5Y cells and primary astrocytes. The anti-amyloid effects of LX2343 were attributed to suppressing JNK-mediated APPThr668 phosphorylation, thus inhibiting APP cleavage on one hand, and inhibiting BACE1 enzymatic activity with an IC50 value of 11.43±0.36 μmol/L, on the other hand. Furthermore, LX2343 acted as a non-ATP competitive PI3K inhibitor to negatively regulate AKT/mTOR signaling, thus promoting autophagy, and increasing Aβ clearance. Administration of LX2343 in APP/PS1 transgenic mice significantly ameliorated cognitive deficits and markedly ameliorated the Aβ pathology in their brains.

Conclusion: LX2343 ameliorates cognitive dysfunction in APP/PS1 transgenic mice via both Aβ production inhibition and clearance promotion, which highlights the potential of LX2343 in the treatment of AD.

Bibliographic Notes:
Li-hong Hu and Xu Shen (CAS Key Laboratory of Receptor Research and State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai and University of Chinese Academy of Sciences, Beijing, China) are corresponding authors on this paper.

Therapeutic Agent

Therapeutic Information:
Therapy Type:
Small Molecule
Therapeutic Agent:
LX2343
Therapeutic Target:
Multi Target

Animal Model

Model Information:
Species:
Mouse
Model Type:
APPxPS1
Strain/Genetic Background:
B6C3

Experimental Design

Is the following information reported in the study?:
Power/Sample Size Calculation
Randomized into Groups
Blinded for Treatment
Blinded for Outcome Measures
Pharmacokinetic Measures
Pharmacodynamic Measures
Toxicology Measures
ADME Measures
Biomarkers
Dose
Formulation
Route of Delivery
Duration of Treatment
Frequency of Administration
Age of Animal at the Beginning of Treatment
Age of Animal at the End of Treatment
Sex as a Biological Variable
Study Balanced for Sex as a Biological Variable
Number of Premature Deaths
Number of Excluded Animals
Statistical Plan
Genetic Background
Inclusion/Exclusion Criteria Included
Conflict of Interest

Outcomes

Outcome Measured
Outcome Parameters
Behavioral
Morris Water Maze
Motor Function
Swimming Speed
Histopathology
beta Amyloid Load
Dense-core/Compact Plaques
Biochemical
A Disintegrin and Metalloproteinase Domain 10 (ADAM10)
Autophagosomal Marker LC3-I
Autophagosomal Marker LC3-II
Protein Kinase B (Akt/PKB)
phospho-Protein Kinase B (phospho-Akt/PKB)
beta-Site Amyloid Precursor Protein Cleaving Enzyme 1 (BACE1)
Brain-beta Amyloid Peptide 40
Brain-beta Amyloid Peptide 42
IC50
c-Jun N-terminal Kinase (JNK)
phospho-c-Jun N-terminal Kinase (phospho-JNK)
Mechanistic Target of Rapamycin (mTOR)
phospho-Mechanistic Target of Rapamycin (phospho-mTOR)
p62/Sequestosome 1 (SQSTM1)
Phosphoinositide 3-Kinase (PI3K)
phospho-Phosphoinositide 3-Kinase (phospho-PI3K)
Postsynaptic Density Protein 95 (PSD95)
Amyloid Precursor Protein (APP)
Soluble Amyloid Precursor Protein beta (sAPP beta)
Synaptophysin
Unc-51 Like Autophagy Activating Kinase 1 (ULK1)
phospho-Unc-51 Like Autophagy Activating Kinase 1 (phospho-ULK1)
Vesicle-Associated Membrane Protein (VAMP)
phospho-p70 S6 Kinase
Immunochemistry
beta Amyloid Load
Autophagosomal Marker LC3-II
Cell Biology
beta Amyloid Peptide Clearance
Cell Viability
Autophagic Markers
Toxicology
Alanine Aminotransferase (ALT)
Aspartate Aminotransferase (AST)
Body Weight
Cell Viability
Liver Function
Mortality
Blood/Serum Clinical Chemistry

Source URL: http://alzped.nia.nih.gov/small-molecule-lx2343