Synthetic studies on the extracellular domain of the T cell immunoreceptor with immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain (TIGIT) using Trt-K10 solubilizing tags

Naoya Iwamoto; Jumpei Sasaki; Saya Ohno; Keisuke Aoki; Yusuke Usui; Shinsuke Inuki; Hiroaki Ohno; Shinya Oishi

doi:10.1016/j.bmc.2023.117585

Synthetic studies on the extracellular domain of the T cell immunoreceptor with immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain (TIGIT) using Trt-K₁₀ solubilizing tags

Bioorg Med Chem. 2024 Feb 1:99:117585. doi: 10.1016/j.bmc.2023.117585. Epub 2023 Dec 30.

Authors

Naoya Iwamoto¹, Jumpei Sasaki¹, Saya Ohno², Keisuke Aoki³, Yusuke Usui¹, Shinsuke Inuki¹, Hiroaki Ohno¹, Shinya Oishi⁴

Affiliations

¹ Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
² Laboratory of Medicinal Chemistry, Kyoto Pharmaceutical University, Yamashina-ku, Kyoto 607-8412, Japan.
³ Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan; Laboratory of Medicinal Chemistry, Kyoto Pharmaceutical University, Yamashina-ku, Kyoto 607-8412, Japan.
⁴ Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan; Laboratory of Medicinal Chemistry, Kyoto Pharmaceutical University, Yamashina-ku, Kyoto 607-8412, Japan. Electronic address: soishi@mb.kyoto-phu.ac.jp.

PMID: 38219557
DOI: 10.1016/j.bmc.2023.117585

Abstract

The T cell immunoreceptor with immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain (TIGIT) is an inhibitory immunoreceptor expressed on lymphocytes that serves as a promising target for cancer immunotherapy. In this study, facile synthetic protocols to produce the extracellular domain of TIGIT were investigated for applications of TIGIT in mirror-image screening. During the synthesis via sequential native chemical ligations, we encountered problems with significantly poor solubility of the ligated products. Introducing trityl-type solubilizing auxiliaries, which also functioned as temporary protecting groups for cysteine residues, facilitated a flexible order of ligations and efficient purification protocols. After refolding under appropriate conditions, the synthetic TIGIT showed a sufficient affinity toward its target ligand CD155.

Keywords: CD155; Chemical protein synthesis; Solubilizing tag; T cell immunoreceptor with immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain; TIGIT.

MeSH terms

Immunoglobulins*
Immunotherapy
Receptors, Immunologic
T-Lymphocytes*
Tyrosine

Substances

Immunoglobulins
Receptors, Immunologic
Tyrosine