Molecular engineering of short half-life small peptides (VIP, αMSH and γ₃MSH) fused to latency-associated peptide results in improved anti-inflammatory therapeutics

Abstract

Incorporation of small anti-inflammatory peptides within the LAP shell and delivered as recombinant protein or through gene therapy can control inflammatory and arthritic disease. This platform delivery can be developed to control human arthritides and other autoimmune diseases.

Latency can be conferred to a variety of cytokines by fusion with latency-associated peptide from TGF-β

“…In addition, LAP- 3 MSH recombinant protein was demonstrated to be more effective in inhibiting urate crystal-induced peritonitis at a 30 fold lower molar concentration than the free  3 MSH peptide. LAP-VIP had a thousand fold longer half-life than free VIP [6].…”

“…We have made fusion proteins between LAP and several therapeutic agents. We replaced the furin cleavage site in LAP with a collagenous matrix metalloproteinase (MMP) cleavage site and replaced TGF- with IFN- [5] or small anti-inflammatory peptides [6]. Metalloproteinases are tightly regulated enzymes involved in development, cell migration and are highly upregulated by pro-inflammatory cytokines and during tissue remodelling [7].…”

“…We have also mutated Cys 33 to Ser to avoid interactions of our fusion proteins with LTBP, a modification that did not affect secretion levels of LAP fusion proteins [6].…”

“…Both were therapeutic in CIA in DBA/1 mice when delivered by intramuscular DNA delivery after onset of disease compared with the empty LAP construct alone [6]. In addition, LAP- 3 MSH recombinant protein was demonstrated to be more effective in inhibiting urate crystal-induced peritonitis at a 30 fold lower molar concentration than the free  3 MSH peptide.…”

A comparative study of matrix metalloproteinase and aggrecanase mediated release of latent cytokines at arthritic joints

“…To overcome the limitations of most cytokines, we created fusion proteins of the latency-associated peptide (LAP) of TGF-β and several therapeutic agents. The furin cleavage site in LAP was replaced with a collagenous matrix metalloproteinase (MMP) cleavage site and TGF-β was replaced with interferon-β (IFN-β) or short anti-inflammatory peptides 3 4. We have also mutated Cys 33 to Ser to avoid interactions of our fusion proteins with latent TGF-β binding protein; this mutation did not affect secretion levels of the LAP fusion proteins 4 5.…”