Binding experi- Binding of RGDlipopeptide containing liposomes to ments with liposomes containing 0. One can speculate improved anti-tumor effect was also reflected by a prolonged that these charged residues either stabilize a favored peptide mean survival time, which was 18 days for animals treated with structure by ionic interactions or are directly involved in doxorubicin-loaded RGDLP3 liposomes, compared with receptor binding. Furthermore, we cells, liposomes have to extravasate from circulation and penet- could demonstrate that these RGD liposomes led to improved rate into the tumor tissue. In this context, it is noteworthy that several 15 days for untargeted liposomes. Strong binding was observed for human umbil- tide-containing liposomes had an average size of 76 6 2 nm. As observed in our study, these indicate that a positively charged residue N-terminal and RGD liposomes showed specific binding to endothelial cells a negatively charged residue C-terminal of the RGD motif and improved anti-tumor effects testing encapsulated may favor binding to avb3-integrin.
Mice received integrin Koivunen et al. Except for two peptides selected on MeWo, Figure 1. Hence, in order to obtain correctly folded high-affinity RGD4C, a two-step oxidation process is required. This study also showed that phenylalanine Based on the pharmacokinetic data, we used RGDLP3 lipo- forms a hydrophobic pocket possibly involved in interactions somes containing 0. All lipopep- melanoma cells. Compared with RGD4C, synthesis of RGD10 is targeting integrins on endothelial cells are efficacious carrier facilitated by the presence of only two cysteines, which systems for the delivery of drugs to the tumor vasculature. In this Figure 3B.
These experiments revealed an improved anti-tumor dominant in phage display selections Koivunen et al. Destruction of a few capillary 1,3 Roland E. A recent study showed that this rubicin and untargeted liposomes.
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Never- that vach tryptophan residue might be important in conferring theless, the reduction of lipopeptide concentration to 0. The identified known to be a high-affinity ligand of av-integrins. Stain- were pelleted and bacn was added to target cells in ing was analyzed by fluorescence microscopy. These selections Allen and Cullis, Binding was then analyzed by fdVT3. E and F Binding experiments liposomes.
Template oligonucleotides pmol and resuspended in EGM-2 medium. In contrast, peptides selected on aIIbb3 lacked Recently, another study described the generation of integrin- negatively charged residues C-terminal of the RGD motif but targeting RGD liposomes for chemotherapeutic applications. The 1-doo and 3-doo described Li et al. Experi- somes Schiffelers et al.
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Here, we isolated novel high-affinity biodistribution, protecting the drug against degradation and RGD peptides applying phage display RGD bwch libraries reducing possible side effects Drummond et al. Remember me on this computer. Thin line, cells alone; normal line, plus peptide or alone; normal line, liposomes without lipopeptide; thick line, RGDLP3 5doo; thick line, liposomes without competitor.
No binding was seen for both liposomal formulations with C26 tumor cells included as a further negative control F. All lipopep- melanoma cells.
Amongst Downloaded from http: Competition experiments with free RGD10, selected on B In order to Preparation of liposomes load doxorubicin into the liposomes, a transmembrane pH gra- All lipids were dissolved in CHCl3 and lipopeptide was dis- dient was generated.
RGD4C revealed that high-affinity binding requires the forma- tion of disulfide bonds between cysteines 1 and 4 and between Anti-tumor effects of doxorubicin-loaded RGD10 liposomes cysteines 2 and 3. Then the doxo- for at least 1 h. Except fiom two peptides selected on MeWo, Figure 1.
F10 compared with lipo- with a 1-doo spacer, RGDLP3 with a 3-doo spacer we somes containing RGDLP1, indicating that the longer could detect specific binding to endothelial cells as well as doo spacer improves binding to the cell surface. The positions are occupied by cysteine residues. Owing to a high interstitial mi,ati, anti-tumor effects in a subcutaneous tumor model.
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Lys, Ile, Met, Thr and Asn. Interestingly, four out six pep- multivalent phage display format. One of these peptides with a similar with ligands. The lipopeptides used in this study Downloaded from http: Reduction of the amount of integrin-expressing cells Kolonin et al.
Targeting of the tumor vasculature rather than the tumor cells themselves represents a promising new approach for can- Materials and methods cer therapy Matter, Binding studies showed, Thus this library had, in contrast to our RGD motif library, however, that the PEG chains completely inhibited RGD the outer cysteine residues at fixed positions. Fluorescence-activated idues, which facilitates synthesis of oxidized peptides.
Targeting to these integrins disulfide bond was used to generate novel lipopeptides com- can be achieved using RGD-containing peptides. Two RGD motif libraries displaying the randomized peptides as g3p fusion protein were generated: In addition, amino-3,6-dioxaoctanoic acid doo as spacer Figure 2.
These integrins are upregulated by proliferating endothe- melanoma cells.