20 kDa PEGylated Adrenomedullin as a New Therapeutic Candidate for Inflammatory Bowel Disease

Shinya Ashizuka,Sayaka Nagata,Masaji Tamura,Taku Yoshiya,Mariko Tokashiki,Kumiko Yoshizawa-Kumagaye,Joji Kato,Goro Miki,Kazuo Kitamura,Motoo Yamasaki,Nobuko Kuroishi

doi:10.3390/gidisord2040033

Shinya Ashizuka, Sayaka Nagata + Show 9 more

Open Access

https://doi.org/10.3390/gidisord2040033

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Abstract

Human adrenomedullin (AM), a hypotensive peptide, also has anti-colitis activity. We prepared a polyethylene glycol (PEG) ylated form of AM through the conjugation of PEG-AM (1–15) and AM (15–52). Highly pure monomeric 20 kDa PEG-AM (20kPEG-AM) stimulated cyclic adenosine monophosphate production in HEK-293 cells stably expressing the type 1 AM receptor in a dose-dependent manner. The half-life of 20kPEG-AM was 7.4 h following subcutaneous administration in mice. We assessed the anti-colitis effect of subcutaneous 20kPEG-AM administration in the dextran sodium sulfate murine colitis model. Single and double subcutaneous injection of 20kPEG-AM significantly reduced total inflammation scores. These results suggest that 20kPEG-AM is a promising therapeutic candidate for the treatment of human inflammatory bowel diseases.

Highlights

Human adrenomedullin (AM), a 52-residue peptide with an amidated C-terminus and an intramolecular disulfide bond, was initially identified as a vasodilatory peptide derived from a human pheochromocytoma [1]
The N-terminal segment without a polyethylene glycol (PEG) moiety was first synthesized by Boc solid-phase peptide synthesis and purified by high-performance liquid chromatography (HPLC)
The N- and C-terminal segments were ligated in the presence of thiophenol to yield reduced PEG20000-AM, which was oxidized with iodine and purified by HPLC to yield 20kPEG-AM

Summary

Introduction

Human adrenomedullin (AM), a 52-residue peptide with an amidated C-terminus and an intramolecular disulfide bond, was initially identified as a vasodilatory peptide derived from a human pheochromocytoma [1]. AM showed therapeutic effects in experimental models of ischemic heart disease, stroke, retinochoroidal disease, and insulin intolerance [4,6,7,8,9]. AM showed therapeutic effects in an experimental model of dextran sulfate sodium (DSS)-induced colitis. AM showed a therapeutic effect in patients with ulcerative colitis and Crohn’s disease following continuous intravenous injection [10,11]. These preclinical and clinical applications of AM required continuous or frequent administration because the plasma half-life of AM is only

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