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Development of a Non-IgG PD-1/PD-L1 Inhibitor by in Silico Mutagenesis and an In-Cell Protein-Protein Interaction Assay.
- Source :
-
ACS chemical biology [ACS Chem Biol] 2021 Feb 19; Vol. 16 (2), pp. 316-323. Date of Electronic Publication: 2021 Jan 15. - Publication Year :
- 2021
-
Abstract
- Inhibiting the programmed death-1 (PD-1)/programmed death ligand 1 (PD-L1) axis by monoclonal antibodies (mAbs) is a successful cancer immunotherapy. However, mAb-based drugs have various disadvantages including high production costs and large molecular sizes, which motivated us to develop a smaller alternative drug. Since PD-L1 binds PD-1 with moderate affinity, a higher affinity PD-1 variant should serve as a competitive inhibitor of the wild-type PD-1/PD-L1 interaction. In this report, we conducted in silico point mutagenesis of PD-1 to identify potent PD-1 variants with a higher affinity toward PD-L1 and refined the in silico results using a luciferase-based in-cell protein-protein interaction (PPI) assay. As a result, a PD-1 variant was developed that had two mutated amino acids (T76Y, A132V), termed 2-PD-1. 2-PD-1 could bind with PD-L1 at a dissociation constant of 12.74 nM. Moreover, 2-PD-1 successfully inhibited the PD-1/PD-L1 interaction with a half maximal inhibitory concentration of 19.15 nM and reactivated the T cell with a half maximal effective concentration of 136.1 nM. These results show that in silico mutagenesis combined with an in-cell PPI assay verification strategy successfully prepared a non-IgG inhibitor of the PD-1/PD-L1 interaction.
- Subjects :
- Computer Simulation
HeLa Cells
Humans
Immune Checkpoint Proteins genetics
Immune Checkpoint Proteins metabolism
Lymphocyte Activation drug effects
Mutagenesis
Point Mutation
Programmed Cell Death 1 Receptor genetics
Protein Engineering
T-Lymphocytes drug effects
B7-H1 Antigen metabolism
Immune Checkpoint Proteins pharmacology
Programmed Cell Death 1 Receptor metabolism
Protein Binding drug effects
Subjects
Details
- Language :
- English
- ISSN :
- 1554-8937
- Volume :
- 16
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- ACS chemical biology
- Publication Type :
- Academic Journal
- Accession number :
- 33448787
- Full Text :
- https://doi.org/10.1021/acschembio.0c00817