PYC pyc therapeutics limited

how to get into the brain, page-5

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    Hi Wayne,

    Further to our discussion re Roche & BBBP's, I came across this technical presentation today which I believe points to the Phylomer BEN_5000 (the focus of additional studies with Roche) as a BBBP. I've highlighted in bold font what caught my attention and am currently researching this some more as it may provide us with a few more clues.

    http://www.phylogica.com/media/PhylogicaTechnologyCapabilities-web-Feb2012.pdf

    Screening for novel Cell-Penetrating Phylomers® (CPPs)

    In collaboration with Roche, Phylogica has searched its Phylomer® libraries for novel cell penetrating peptides that can be used to deliver therapeutic cargoes inside cells. Phage displayed Phylomer® libraries were screened for the ability to internalise into whole mammalian cells, which yielded almost 1000 unique hits. A subset of 186 synthetic and recombinant peptides, were then assessed using a combination of FACS and live cell confocal microscopy to confirm cell-penetrating activity (Figure 12). A total of 17 novel cell-penetrating peptides were identified corresponding to a functional hit-rate of 11%.

    Notably, multiple classes of CPPs were identified, reflecting the high levels of structural and functional diversity found within the Phylomer® libraries. These included the well characterised TAT-like class of cationic amphipathic peptides, as well as bacterial virulence factors known to be involved in cell invasion and also a panel of novel sequences not previously associated with cell penetration. In addition, several CPPs were shown to only internalise selectively into certain cell types, raising the possibility that therapeutic cargoes could be delivered in a cell-specific manner (Figure 13).

    Of particular interest were two novel peptides derived from fragments of natural proteins, which were shown to selectively internalise into a murine brain endothelial cell-line (bEnd.3). These peptides are now the subject of ongoing studies to determine their ability to penetrate across the blood brain barrier. All Phylomer® CPPs were found to be non-toxic at concentrations up to 50µM when tested using a cell-based fluorescent vital dye assay, which is in vast excess of their minimal concentration required for functional activity.

    Regards,
    Tony




 
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