AEGiS-13IAC: A potent anti-HIV compound produced through knowledge-based design of a phage display selection strategy.

13th International AIDS Conference

Durban, South Africa - July 9-July 14, 2000

A potent anti-HIV compound produced through knowledge-based design of a phage display selection strategy.

Int Conf AIDS 2000 Jul 9-14; 13:39 (abstract no.. LbPeA7029)

Gorochov G, Hartley O, Dorgham K, Perez-Bercaff D, David A, Gaertner H, Offord RE, Pancino G, Debre P
Faculty of Paris, Hopital Pitie-Salpetriere, Paris, France. Fax: +33 1 42 17 74 90, E-mail: guy.gorochov@psl.ap-hop-paris.fr.

BACKGROUND: The chemokine receptors CCR5 and CXCR4 are promising targets for HIV therapy. We have used a phage display aproach 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 introduced diversity into the 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. An original selection strategy on CCR5-expressing cells was used.

RESULTS: 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. 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.

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.

Keywords: AEGIS, HIV Infections, HIV-1, RANTES, Receptors, Chemokine, HIV Seropositivity, Chemokines, Macrophages, AIDS
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