Synthesis and Pharmacological Characterization of Visabron, a Backbone Cyclic Peptide Dual Antagonist of α4β1 (VLA-4)/α9β1 Integrin for Therapy of Multiple Sclerosis.

Adi Lahiani,Cezary Marcinkiewicz,Philip Lazarovici,Michal Klazas,Shira Merzbach,Erik M. Schaefer,Amnon Hoffman,Adi Schumacher-Klinger,Haim Ovadia,Johnny N. Naoum,Chaim Gilon,Limor Rubin,Susan Cornell-Kennon,Jehoshua Katzhendler

doi:10.1021/jacsau.1c00496

Adi Lahiani, Cezary Marcinkiewicz + Show 12 more

Open Access

https://doi.org/10.1021/jacsau.1c00496

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Abstract

Integrins α4β1/ α9β1 are important in the pathogenesis and progression of inflammatory and autoimmune diseases by their roles in leukocyte activation and trafficking. Natalizumab, a monoclonal antibody selectively targeting α4β1 integrin and blocking leukocyte trafficking to the central nervous system, is an immunotherapy for multiple sclerosis (MS). However, due to its adverse effects associated with chronic treatment, alternative strategies using small peptide mimetic inhibitors are being sought. In the present study, we synthesized and characterized visabron c (4–4), a backbone cyclic octapeptide based on the sequence TMLD, a non-RGD unique α4β1 integrin recognition sequence motif derived from visabres, a proteinous disintegrin from the viper venom. Visabron c (4–4) was selected from a minilibrary with conformational diversity based on its potency and selectivity in functional adhesion cellular assays. Visabron c (4–4)’s serum stability, pharmacokinetics, and therapeutic effects following ip injection were assessed in an experimental autoimmune encephalomyelitis (EAE) animal model. Furthermore, visabron c (4–4)’s lack of toxic effects in mice was verified by blood analysis, tissue pathology, immunogenicity, and “off-target” effects, indicating its significant tolerability and lack of immunogenicity. Visabron c (4–4) can be delivered systemically. The in vitro and in vivo data justify visabron c (4–4) as a safe alternative peptidomimetic lead compound/drug to monoclonal anti-α4 integrin antibodies, steroids, and other immunosuppressant drugs. Moreover, visabron c (4–4) design may pave the way for developing new therapies for a variety of other inflammatory and/or autoimmune diseases.

Highlights

Integrins are a family of cell surface receptors composed of noncovalently linked α and β subunits that regulate many biological functions in eukaryotic cells and play essential roles in regulating cell adhesion and migration.[1]
Pubs.acs.org/jacsau study, we describe the isolation of visabres disintegrin from viper venom, identification of the TMLD motif selective for α4β1/α4β7 integrin inhibition, and attempts for its cyclization to generate visabron, a backbone cyclic peptide dual antagonist of α4β1 (VLA-4)/ α9β1 integrin with druglike properties
Mass spectroscopy analyses of this fraction indicated it is enriched in disintegrins but still contaminated with other venom proteins (Table S3)

Summary

■ INTRODUCTION

Integrins are a family of cell surface receptors composed of noncovalently linked α and β subunits that regulate many biological functions in eukaryotic cells and play essential roles in regulating cell adhesion and migration.[1]. The α4β1 and α4β7 integrins have been established as the most selective receptors for fibronectin, vascular adhesion molecule (VCAM-1), and mucosal addressin cell adhesion molecule-1 (MdCAM-1) and have been recognized as important targets for the development of monoclonal antibodies, peptidomimetics, and small molecules for therapy of multiple sclerosis and inflammatory diseases of the digestive system.[2,3] The α4β1 ( known as very late antigen-4 (VLA-4) or CD49d/CD29), α4β7, and α9β1 integrins bind to an acidic motif termed “LDV” in fibronectin They share several structural and functional properties, have similar tissue expression profiles, and are classified into a common subgroup within the integrin family.

■ RESULTS AND DISCUSSION

■ REFERENCES

■ ACKNOWLEDGMENTS