Synthesis and characterization of a thermally-responsive tumor necrosis factor antagonist.
Numerous antagonists of tumor necrosis factor alpha (TNFalpha) have been developed to attenuate inflammation and accompanying pain in many disease processes. Soluble TNF receptor type II (sTNFRII) is one such antagonist that sequesters TNFalpha away from target receptors and attenuates its activity. Systemic delivery of soluble TNF receptors or other antagonists may have deleterious side effects associated with immune suppression, so that strategies for locally targeted drug delivery are of interest. Elastin-like polypeptides (ELPs) are biopolymers capable of in situ drug depot formation through thermally-driven supramolecular complexes at physiological temperatures. A recombinant fusion protein between ELP and sTNFRII was designed and evaluated for retention of bivalent functionality. Thermal sensitivity was observed by formation of supramolecular submicron-sized particles at 32 degrees C, with gradual resolubilization from the depot observed at physiological temperatures. In vitro refolding of the sTNFRII domain was required and the purified product exhibited an equilibrium dissociation constant for interacting with TNFalpha that was seven-fold higher than free sTNFRII. Furthermore, anti-TNF activity was observed in inhibiting TNFalpha-mediated cytotoxicity in the murine L929 fibrosarcoma assay. Potential advantages of this ELP-sTNFRII fusion protein as an anti-TNFa drug depot include facility of injection, in situ depot formation, low endotoxin content, and functionality against TNFalpha.
Department of Biomedical Engineering, Duke University, Durham, NC, USA; Division of Neurosurgery, The Ottawa Hospital, Ottawa, ON, Canada.
Mohammed F
MF
Shamji
Jun
J
Chen
Allan H
AH
Friedman
William J
WJ
Richardson
Ashutosh
A
Chilkoti
Lori A
LA
Setton
D000818:Animals; D004549:Elastin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005354:Fibrosarcoma / Q000378:metabolism / Q000473:pathology; D006801:Humans; D020128:Inhibitory Concentration 50; D007700:Kinetics; D007739:L Cells; D051379:Mice; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D020673:Protein Renaturation; D017434:Protein Structure, Tertiary; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012995:Solubility; D013696:Temperature; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000138:chemical synthesis* / Q000737:chemistry
D010455:Peptides; D047889:Receptors, Tumor Necrosis Factor, Type II; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D004549:Elastin