PMID- 17968946
OWN - NLM
STAT- MEDLINE
DCOM- 20071219
LR  - 20141120
IS  - 0004-3591 (Print)
IS  - 0004-3591 (Linking)
VI  - 56
IP  - 11
DP  - 2007 Nov
TI  - 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.
PG  - 3650-61
AB  - 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.
FAU - Shamji, Mohammed F
AU  - Shamji MF
AD  - Duke University, Durham, North Carolina 27708, USA.
FAU - Betre, Helawe
AU  - Betre H
FAU - Kraus, Virginia B
AU  - Kraus VB
FAU - Chen, Jun
AU  - Chen J
FAU - Chilkoti, Ashutosh
AU  - Chilkoti A
FAU - Pichika, Rajeswari
AU  - Pichika R
FAU - Masuda, Koichi
AU  - Masuda K
FAU - Setton, Lori A
AU  - Setton LA
LA  - eng
GR  - R01-EB0-002263/EB/NIBIB NIH HHS/United States
GR  - R21-AR-052745/AR/NIAMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Arthritis Rheum
JT  - Arthritis and rheumatism
JID - 0370605
RN  - 0 (Anti-Inflammatory Agents)
RN  - 0 (Interleukin 1 Receptor Antagonist Protein)
RN  - 0 (Peptides)
RN  - 0 (Receptors, Interleukin-1)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 9007-58-3 (Elastin)
SB  - IM
MH  - Animals
MH  - Anti-Inflammatory Agents/*pharmacokinetics
MH  - Cell Division/drug effects/immunology
MH  - Chondrocytes/cytology/drug effects/immunology
MH  - Drug Delivery Systems/methods
MH  - Drug Design
MH  - Elastin/*genetics
MH  - Humans
MH  - In Vitro Techniques
MH  - Interleukin 1 Receptor Antagonist Protein/*genetics
MH  - Intervertebral Disc/cytology
MH  - Lymphocytes/cytology/immunology
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Peptides/genetics
MH  - Receptors, Interleukin-1/metabolism
MH  - Recombinant Fusion Proteins/*genetics/*pharmacokinetics
MH  - Temperature
MH  - Thymus Gland/cytology
MH  - U937 Cells
EDAT- 2007/10/31 09:00
MHDA- 2007/12/20 09:00
CRDT- 2007/10/31 09:00
PHST- 2007/10/31 09:00 [pubmed]
PHST- 2007/12/20 09:00 [medline]
PHST- 2007/10/31 09:00 [entrez]
AID - 10.1002/art.22952 [doi]
PST - ppublish
SO  - Arthritis Rheum. 2007 Nov;56(11):3650-61. doi: 10.1002/art.22952.