title: PEGylation extends circulation half-life while preserving in vitro and in vivo activity of tissue inhibitor of metalloproteinases-1 (TIMP-1).
abstract: Excess proteolytic activity of matrix metalloproteinases (MMPs) contributes to the development of arthritis, cardiovascular diseases and cancer progression, implicating these enzymes as therapeutic targets. While many small molecule inhibitors of MMPs have been developed, clinical uses have been limited, in part by toxicity and off-target effects. Development of the endogenous tissue inhibitors of metalloproteinases (TIMPs) as recombinant biopharmaceuticals represents an alternative therapeutic approach; however, the short plasma half-life of recombinant TIMPs has restricted their potential in this arena. To overcome this limitation, we have modified recombinant human TIMP-1 (rhTIMP-1) by PEGylation on lysine residues. We analyzed a mixture of mono- and di-PEGylated rhTIMP-1 species modified by attachment of 20 kDa mPEG chains (PEG(20K)-TIMP-1), as confirmed by SELDI-TOF mass spectrometry. This preparation retained complete inhibitory activity toward the MMP-3 catalytic domain and partial inhibitory activity toward full length MMP-9. Pharmacokinetic evaluation showed that PEGylation extended the plasma half-life of rhTIMP-1 in mice from 1.1 h to 28 h. In biological assays, PEG(20K)-TIMP-1 inhibited both MMP-dependent cancer cell invasion and tumor cell associated gelatinase activity. Overall these results suggest that PEGylated TIMP-1 exhibits improved potential for development as an anti-cancer recombinant protein therapeutic, and additionally may offer potential for clinical applications in the treatment of other diseases.
authors.affiliation: Department of Cancer Biology, Mayo Clinic Cancer Center, Jacksonville, Florida, United States of America.
authors.forename: Jyotica
authors.initials: J
authors.lastname: Batra
authors.forename: Jessica
authors.initials: J
authors.lastname: Robinson
authors.forename: Christine
authors.initials: C
authors.lastname: Mehner
authors.forename: Alexandra
authors.initials: A
authors.lastname: Hockla
authors.forename: Erin
authors.initials: E
authors.lastname: Miller
authors.forename: Derek C
authors.initials: DC
authors.lastname: Radisky
authors.forename: Evette S
authors.initials: ES
authors.lastname: Radisky
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D018093:Gelatinases / Q000378:metabolism; D006207:Half-Life; D006801:Humans; D008239:Lysine / Q000737:chemistry*; D019278:Matrix Metalloproteinase 3 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D061965:Matrix Metalloproteinase Inhibitors / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D009361:Neoplasm Invasiveness; D011092:Polyethylene Glycols; D011480:Protease Inhibitors / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D061965:Matrix Metalloproteinase Inhibitors; D011480:Protease Inhibitors; D011994:Recombinant Proteins; C560822:TIMP1 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1; D011092:Polyethylene Glycols; D018093:Gelatinases; D019278:Matrix Metalloproteinase 3; D020780:Matrix Metalloproteinase 9; D008239:Lysine