A-β immunogenic peptide carrier conjugates and methods of producing same

摘要

The present invention is directed to methods of producing conjugates of Aβ peptide immunogens with protein/polypeptide carrier molecules, which are useful as immunogens, wherein peptide immunogens are conjugated to protein carriers via activated functional groups on amino acid residues of the carrier or of the optionally attached linker molecule, and wherein any unconjugated reactive functional groups on amino acid residues are inactivated via capping, thus retaining the immunological functionality of the carrier molecule, but reducing the propensity for undesirable reactions that could render the conjugate less safe or effective. Furthermore, the invention also relates to such immunogenic products and immunogenic compositions containing such immunogenic products made by such methods.

主权项

1. A method of producing an immunogenic conjugate, the method comprising the steps of:
(a) derivatizing one or more functional groups of a carrier protein to generate an activated functional group on the carrier protein; (b) reacting a peptide immunogen comprising an Aβ fragment with the carrier protein of step (a) under conditions to form a conjugate, wherein the Aβ fragment comprises residues 16-23 of Aβ (SEQ ID NO:21) and the peptide immunogen is covalently attached to the activated functional group on the carrier protein; and (c) further reacting the conjugate of step (b) with N-acetylcysteamine to inactivate any remaining activated functional group on the carrier protein to generate the immunogenic conjugate, wherein a capping molecule, formed by reaction of the conjugate with the N-acetylcysteamine, is covalently attached to the remaining activated functional group on the carrier protein, wherein the immunogenic conjugate has the formula: wherein, C is a CRM197 carrier protein, Xd is a derivatized functional group of an amino acid residue of the carrier protein, P is the peptide immunogen comprising the Aβ fragment covalently attached to the derivatized functional group of the amino acid residue of the carrier protein, R is the capping molecule covalently attached to the derivatized functional group of an amino acid residue of the carrier protein, wherein the functionality of the carrier protein is preserved such that it retains its ability to elicit the desired immune response against the peptide immunogen that would otherwise not occur without a carrier, n is an integer greater than 0, but less than or equal to 38, and p is an integer greater than 0, but less than or equal to 38.