Surface plasmon resonance and cytotoxicity assays of drug efficacies predicted computationally to inhibit p53/MDM2 interaction

Xiaoying Wang; Patrycja Magdziarz; Ernest Enriquez; Wang Zhao; Chris Quan; Narek Darabedian; Jamil Momand; Feimeng Zhou

doi:10.1016/j.ab.2019.01.012

Surface plasmon resonance and cytotoxicity assays of drug efficacies predicted computationally to inhibit p53/MDM2 interaction

Anal Biochem. 2019 Mar 15:569:53-58. doi: 10.1016/j.ab.2019.01.012. Epub 2019 Feb 2.

Authors

Xiaoying Wang¹, Patrycja Magdziarz², Ernest Enriquez², Wang Zhao², Chris Quan², Narek Darabedian², Jamil Momand³, Feimeng Zhou⁴

Affiliations

¹ College of Chemistry and Chemical Engineering, Central South University, Changsha, Hunan, 410083, PR China.
² Department of Chemistry and Biochemistry, California State University, Los Angeles, Los Angeles, CA, 90032, USA.
³ Department of Chemistry and Biochemistry, California State University, Los Angeles, Los Angeles, CA, 90032, USA. Electronic address: JMomand@calstatela.edu.
⁴ Department of Chemistry and Biochemistry, California State University, Los Angeles, Los Angeles, CA, 90032, USA. Electronic address: fzhou@calstatela.edu.

Abstract

Docking on the p53-binding site of murine double minute 2 (MDM2) by small molecules restores p53's tumor-suppressor function. We previously assessed 3244 FDA-approved drugs via "computational conformer selection" for inhibiting MDM2 and p53 interaction. Here, we developed a surface plasmon resonance method to experimentally confirm the inhibitory effects of the known MDM2 inhibitor, nutlin-3a, and two drug candidates predicted by our computational method. This p53/MDM2 interaction displayed a dosage-dependent weakening when MDM2 is pre-mixed with drug candidates. The inhibition efficiency order is nutlin-3a (IC₅₀ = 97 nM) > bepridil (206 nM) > azelastine (307 nM). Furthermore, we verified their anti-proliferation effects on SJSA-1 (wild-type p53 and overexpressed MDM2), SW480 (mutated p53), and SaOs-2 (deleted p53) cancer cell lines. The inhibitory order towards SJSA-1 cell line is nutlin-3a (IC₅₀ = 0.8 μM) > bepridil (23 μM) > azelastine (25 μM). Our experimental results are in line with the computational prediction, and the higher IC₅₀ values from the cell-based assays are due to the requirement of higher drug concentrations to penetrate cell membranes. The anti-proliferation effects of bepridil and azelastine on the cell lines with mutated and deleted p53 implied some p53-independent anti-proliferation effects.

Keywords: Drug screening; Inhibitory effects; MDM2; Repurposing of existing drugs; p53.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Bepridil / chemistry
Bepridil / metabolism
Cell Line, Tumor
Humans
Imidazoles / chemistry
Imidazoles / metabolism
Piperazines / chemistry
Piperazines / metabolism
Protein Binding
Proto-Oncogene Proteins c-mdm2 / antagonists & inhibitors
Proto-Oncogene Proteins c-mdm2 / genetics
Proto-Oncogene Proteins c-mdm2 / metabolism*
Small Molecule Libraries / chemistry*
Small Molecule Libraries / metabolism
Surface Plasmon Resonance*
Tumor Suppressor Protein p53 / antagonists & inhibitors
Tumor Suppressor Protein p53 / genetics
Tumor Suppressor Protein p53 / metabolism*

Substances

Imidazoles
Piperazines
Small Molecule Libraries
Tumor Suppressor Protein p53
nutlin 3
Bepridil
Proto-Oncogene Proteins c-mdm2

Grants and funding

R01 GM105898/GM/NIGMS NIH HHS/United States