Binding of novel peptide inhibitors of type IV collagenases to phospholipid membranes and use in liposome targeting to tumor cells in vitro.
We have recently described a novel cyclic peptide inhibitor CTTHWGFTLC (CTT) for matrix metalloproteinases (MMP)-2 and MMP-9, also called type IV collagenases or gelatinases (E. Koivunen et al., NAT: BIOTECHNOL:, 17: 768-774, 1999). As indicated by its amino acid composition, CTT is hydrophobic, and its partitioning into phospholipid films could be verified by the monolayer technique. Augmented fluorescence emission anisotropy (from 0.064 to 0.349) and reduced collisional quenching by I(-) of the Trp residue in CTT was evident in the presence of unilamellar phosphatidylcholine/phosphatidylethanolamine liposomes, revealing the association of CTT with the lipid bilayers. Gelatinases are potential targets of therapeutic intervention in cancer, and inhibitors of these enzymes can prevent tumor progression in animal models. CTT enhanced 3- to 4-fold the cellular uptake of liposome-encapsulated water-soluble fluorescent marker, rhodamine B by gelatinase-expressing cells. Gelatinase targeting seems to be essential, as modified peptides that were less potent gelatinase inhibitors were also less efficient in promoting the cellular uptake of liposomes. Augmented killing ( approximately 4-fold) of U937 leukemia and HT1080 sarcoma cells was obtained by the CTT-enhanced delivery of Adriamycin-containing liposomes, compared with control liposomes administered without the peptide. These results suggest a novel type of utility for small gelatinase inhibitors in targeted cancer therapy.
Medina
O P
OP
Helsinki Biophysics and Biomembrane Group, Department of Medical Chemistry, Institute of Biomedicine, FIN-00014 University of Helsinki, Helsinki, Finland.
Söderlund
T
T
Laakkonen
L J
LJ
Tuominen
E K
EK
Koivunen
E
E
Kinnunen
P K
PK
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D004791:Enzyme Inhibitors / Q000378:metabolism* / Q000493:pharmacokinetics / Q000494:pharmacology; D006801:Humans; D008081:Liposomes / Q000378:metabolism / Q000493:pharmacokinetics*; D061965:Matrix Metalloproteinase Inhibitors; D009842:Oligopeptides / Q000378:metabolism* / Q000494:pharmacology; D010456:Peptides, Cyclic / Q000378:metabolism / Q000493:pharmacokinetics / Q000494:pharmacology; D010743:Phospholipids / Q000378:metabolism*; D020298:U937 Cells
C415426:CTTHWGFTLC peptide; D004791:Enzyme Inhibitors; D008081:Liposomes; D061965:Matrix Metalloproteinase Inhibitors; D009842:Oligopeptides; D010456:Peptides, Cyclic; D010743:Phospholipids