115. DEVELOPMENT OF POTENT AND STABLE PC6 INHIBITORS TO BLOCK EMBRYO IMPLANTATION FOR FEMALE CONTRACEPTION AND PREVENTION OF HIV

Abstract

Proprotein convertase (PC) 6, a member of the PC family that activate precursor proteins into their active forms, is a critical endometrial factor for embryo implantation. Blocking PC6 production in mice inhibits decidualisation (a critical process of implantation) and inhibits implantation. PCs including PC6 also play a critical role in HIV infection through cleaving HIV envelope precursor protein gp160 into functional gp120 and gp41. PC inhibitors are demonstrated to inhibit HIV transmission via blocking gp160 cleavage. We hypothesised that PC6 is a potential target for the development of female contraception that could also provide protection from HIV infection. One key requirement to prove this concept in an animal model is a potent PC6 inhibitor that is stable in serum. Polyarginine peptide (polyR) is published to be a potent PC6 inhibitor that also inhibits HIV. We have confirmed that polyR inhibits PC6 in vivo and completely blocks decidualisation of human endometrial stromal cells in culture. However, polyR has short serum half-life. The aim of this current study was to generate polyR derivatives that are potent PC6 inhibitors, with increased serum stability. We modified polyR by either PEGylation with different sized PEG (polyethylene glycol) or cyclization, and tested their potency, stability and utility in vivo. Modifications at both terminals of polyR dramatically reduced its PC6 inhibitory potency. PEGylation at the C-terminal, regardless of the PEG size, had no effect. In silico docking experiments showed that N-terminal PEGylation or cyclization affected the binding of polyR to the PC6 active site, but not C-terminal PEGylation. One of the polyR derivatives, C-30kDa-PEG polyR was confirmed to be as potent as the parental peptide but much more stable in serum. Studies are currently in place to inhibit embryo implantation in mice using C-30kDa-PEG polyR and to determine its ability to inhibit HIV.