SSAA09E2

Cell lines

293T cells (transiently transfected with pCDNA 3.1-ACE2), TZM-bl cells (stably expressing luciferase under the control of the HIV-1 promoter and transfected with pcDNA 3.1-ACE2 plasmid)

Preparation Method

293T cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100units/ml penicillin, and 10μg streptomycin. For binding inhibition assays, approximately 1×10⁶ ACE2-transfected 293T cells were detached, washed, and incubated with various concentrations of SSAA09E2 on ice for 30 minutes, followed by addition of purified SARS-CoV spike receptor binding domain-Fc (RBD-Fc) and incubation on ice for 1 hour. For fusion assays, TZM-bl cells expressing ACE2 were overlaid with 293T cells expressing SARS-S envelope and Tat protein, and SSAA09E2 was added either 45 minutes prior to or 2 hours after the overlay.

Reaction Conditions

0-20μM; 30 minutes pre-incubation

Applications

SSAA09E2 significantly inhibited the binding of SARS-S RBD to ACE2 in a dose-dependent manner as measured by flow cytometry. SSAA09E2 also inhibited SARS/HIV pseudotyped virus entry into ACE2-expressing 293T cells with an EC₅₀ of 3.5 ± 0.5μM. In quantitative cell-to-cell fusion assays, SSAA09E2 inhibited fusion when added prior to overlay of SARS-S expressing cells with ACE2-expressing cells.

References:
[1] París-Coderch L, Soriano A, Jiménez C, et al. The antitumour drug ABTL0812 impairs neuroblastoma growth through endoplasmic reticulum stress-mediated autophagy and apoptosis. Cell Death Dis. 2020 Sep 17;11(9):773.