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Abstract
POTENT ANTI-HIV CHEMOKINE ANALOGUES PRODUCED THROUGH A PHAGE DISPLAY SELECTION STRATEGY.
GOROCHOV G, HARTLEY O, OORGHAM K, PANCINO G, DEBRE P, OFFORD R
Background: the chemokine receptors CCR5 and CXCR4 are promising targets for HIV therapy. We have used a phage display approach to isolate mutant forms of RANTES with antiviral activity considerably in excess of the native sequence.
Methods: the mutants were selected from a library whose design was informed by existing models of chemokine structure and activity. (1) We decided to introduce diversity into sequence encoding the N-terminus of the protein, a region known to be involved in the activation of receptors by bound chemokines. (2) The mutant sequences were also extended mimicking the N-terminal extension seen in the previously described inhibitors. (3) The mutant proteins were fused to the phage gene 3 protein via their C-termini, as this region is not believed to play a crucial role in chemokine-receptor interaction. An original selection strategy on CCR5-expressing cells was used. Proteins encoded by two of the most abundant selected sequences (S1 and S2) were prepared by total chemical synthesis, and their activity towards CCR5 was determined.
Results: S2 induces surface down-modulation of, and inhibits viral coat-mediated cell fusion (IC50 of 300 pM). S2 is more potent than (AOP)-RANTES and consistently achieves complete protection of activated lymphocytes and macrophages against primary HIV-1 strains at a concentration of 10 nM. Results obtained from second generation libraries will also be presented.
Conclusions: Using a limited knowledge of the mechanistic and structure-activity factors that determine the anti-HIV action of RANTES analogues, it was possible to tailor a library design and selection strategy capable of isolating one of the most potent anti-HIV chemokine analogue so far described. This technique might be applicable to the discovery of novel inhibitors of other G protein coupled receptors of pharmacological interest.
The 1st. IAS Conference on HIV Pathogenesis and Treatment
Abstract no.
74
Suggested Citation
"GOROCHOVG, et al.
POTENT ANTI-HIV CHEMOKINE ANALOGUES PRODUCED THROUGH A PHAGE DISPLAY SELECTION STRATEGY..
Oral Presentation:
The 1st. IAS Conference on HIV Pathogenesis and Treatment
:
Abstract no.
74"
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