TY - JOUR
T1 - Synthesis and characterization of a thermally-responsive tumor necrosis factor antagonist
AU - Shamji, Mohammed F.
AU - Chen, Jun
AU - Friedman, Allan H.
AU - Richardson, William J.
AU - Chilkoti, Ashutosh
AU - Setton, Lori A.
N1 - Funding Information:
This work was funded by NIH R01EB002263 (LAS), NIH R21AR052745 (LAS), NIH R01GM061232 (AC), and a Pratt-Gardner Predoctoral Research Fellowship (MFS).
PY - 2008/8/7
Y1 - 2008/8/7
N2 - Numerous antagonists of tumor necrosis factor alpha (TNFα) 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 TNFα 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 °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 TNFα that was seven-fold higher than free sTNFRII. Furthermore, anti-TNF activity was observed in inhibiting TNFα-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 TNFα.
AB - Numerous antagonists of tumor necrosis factor alpha (TNFα) 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 TNFα 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 °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 TNFα that was seven-fold higher than free sTNFRII. Furthermore, anti-TNF activity was observed in inhibiting TNFα-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 TNFα.
KW - Drug depot
KW - Elastin-like polypeptide
KW - Fusion protein
KW - Soluble tumor necrosis factor receptor
KW - Tumor necrosis factor alpha
UR - https://www.scopus.com/pages/publications/47349085363
U2 - 10.1016/j.jconrel.2008.04.021
DO - 10.1016/j.jconrel.2008.04.021
M3 - Article
C2 - 18547669
AN - SCOPUS:47349085363
SN - 0168-3659
VL - 129
SP - 179
EP - 186
JO - Journal of Controlled Release
JF - Journal of Controlled Release
IS - 3
ER -