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Development and characterization of a fusion protein between thermally responsive elastin-like polypeptide and interleukin-1 receptor antagonist: sustained release of a local antiinflammatory therapeutic.

Publication ,  Journal Article
Shamji, MF; Betre, H; Kraus, VB; Chen, J; Chilkoti, A; Pichika, R; Masuda, K; Setton, LA
Published in: Arthritis Rheum
November 2007

OBJECTIVE: Interleukin-1 receptor antagonist (IL-1Ra) has been evaluated for the intraarticular treatment of osteoarthritis. Such administration of proteins may have limited utility because of their rapid clearance and short half-life in the joint. The fusion of a drug to elastin-like polypeptides (ELPs) promotes the formation of aggregating particles that form a "drug depot" at physiologic temperatures, a phenomenon intended to prolong the presence of the drug. The purpose of this study was to develop an injectable drug depot composed of IL-1Ra and ELP domains and to evaluate the properties and bioactivity of the recombinant ELP-IL-1Ra fusion protein. METHODS: Fusion proteins between IL-1Ra and 2 distinct sequences and molecular weights of ELP were overexpressed in Escherichia coli. Environmental sensitivity was demonstrated by turbidity and dynamic light scattering as a function of temperature. IL-1Ra domain activity was evaluated by surface plasmon resonance, and in vitro antagonism of IL-1-mediated lymphocyte and thymocyte proliferation, as well as IL-1-induced tumor necrosis factor alpha (TNFalpha) expression and matrix metalloproteinase 3 (MMP-3) and ADAMTS-4 messenger RNA expression in human intervertebral disc fibrochondrocytes. IL-1Ra immunoreactivity was assessed before and after proteolytic degradation of the ELP partner. RESULTS: Both fusion proteins underwent supramolecular aggregation at subphysiologic temperatures and slowly resolubilized at 37 degrees C. Interaction with IL-1 receptor was slower in association but equivalent in dissociation as compared with the commercial antagonist. Anti-IL-1 activity was demonstrated by inhibition of lymphocyte and thymocyte proliferation and by decreased TNFalpha expression and ADAMTS-4 and MMP-3 transcription by fibrochondrocytes. ELP domain proteolysis liberated a peptide of comparable size and immunoreactivity as the commercial IL-1Ra. This peptide was more bioactive against lymphocyte proliferation, nearly equivalent to the commercial antagonist. CONCLUSION: The ELP-IL-1Ra fusion protein proved to retain the characteristic ELP inverse phase-transitioning behavior as well as the bioactivity of the IL-1Ra domain. This technology represents a novel drug carrier designed to prolong the presence of bioactive peptides following intraarticular delivery.

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Published In

Arthritis Rheum

DOI

ISSN

0004-3591

Publication Date

November 2007

Volume

56

Issue

11

Start / End Page

3650 / 3661

Location

United States

Related Subject Headings

  • U937 Cells
  • Thymus Gland
  • Temperature
  • Recombinant Fusion Proteins
  • Receptors, Interleukin-1
  • Peptides
  • Mice, Inbred C57BL
  • Mice
  • Lymphocytes
  • Intervertebral Disc
 

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Shamji, M. F., Betre, H., Kraus, V. B., Chen, J., Chilkoti, A., Pichika, R., … Setton, L. A. (2007). Development and characterization of a fusion protein between thermally responsive elastin-like polypeptide and interleukin-1 receptor antagonist: sustained release of a local antiinflammatory therapeutic. Arthritis Rheum, 56(11), 3650–3661. https://doi.org/10.1002/art.22952
Shamji, Mohammed F., Helawe Betre, Virginia B. Kraus, Jun Chen, Ashutosh Chilkoti, Rajeswari Pichika, Koichi Masuda, and Lori A. Setton. “Development and characterization of a fusion protein between thermally responsive elastin-like polypeptide and interleukin-1 receptor antagonist: sustained release of a local antiinflammatory therapeutic.Arthritis Rheum 56, no. 11 (November 2007): 3650–61. https://doi.org/10.1002/art.22952.
Journal cover image

Published In

Arthritis Rheum

DOI

ISSN

0004-3591

Publication Date

November 2007

Volume

56

Issue

11

Start / End Page

3650 / 3661

Location

United States

Related Subject Headings

  • U937 Cells
  • Thymus Gland
  • Temperature
  • Recombinant Fusion Proteins
  • Receptors, Interleukin-1
  • Peptides
  • Mice, Inbred C57BL
  • Mice
  • Lymphocytes
  • Intervertebral Disc