Methods and compositions for liposomal formulation of antigens and uses thereof
Inventors
Fujii, Gary • Szoka, Jr., Francis C. • WATSON, Douglas S.
Assignees
Molecular Express Inc • University of California San Diego UCSD
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Publication Number
US-9364563-B2
Publication Date
2016-06-14
Expiration Date
Abstract
The present invention relates to liposomal vaccine compositions, methods for the manufacture thereof, and methods for the use thereof to stimulate an immune response in an animal. These compositions comprise dimyristoylphosphatidylcholine (“DMPC”); either dimyristoylphosphatidylglycerol (“DMPG”) or dimyristoyltrimethylammonium propane (“DMTAP”) or both DMPC and DMTAP; and at least one sterol derivative providing a covalent anchor for one or more immunogenic polypeptide(s) or carbohydrate(s).
Core Innovation
The disclosed subject matter relates to liposomal vaccine compositions and a method for preparing compositions comprising one or more immunogenic polypeptides of interest. The method combines dimyristoylphosphatidylcholine (DMPC) with one or more lipids selected from dimyristoylphosphatidylglycerol (DMPG) and dimyristoyltrimethylammonium propane (DMTAP) to provide a lipid mixture, and at least one sterol derivative is included in the lipid mixture to provide a lipid mixture suitable for liposome preparation.
Liposomes are prepared from the lipid mixture, and one or more immunogenic polypeptides are covalently coupled to the at least one sterol derivative. The covalent linkage links the one or more immunogenic polypeptides or carbohydrate(s) to between 1% and 100% of the at least one sterol derivative, and the sterol derivatives function as covalent anchors for immunogenic polypeptides and/or carbohydrates.
The sterol-derivative classes and reactive-linker options are described, including cholesterol derivatives and sterol derivatives with functional moieties suitable for covalent coupling. The disclosure also specifies reactive groups such as amine-reactive, sulfhydryl-reactive, carboxyl-reactive, photoaffinity-reactive, arginine linking group, and carbonyl reactive groups, and an (alkylene oxide)n linker with an average length n between 40 and 1000.
Claims Coverage
The document provides one independent claim directed to a method of preparing immunogen-coupled liposomal compositions, with multiple inventive refinements described in dependent claims, including optional immunostimulatory components, covalent-linkage attachment specifics, quantitative linkage fraction constraints, and constraints on linkage chemistry and/or processing used to form the liposomes.
Sterol-derivative anchored immunogenic polypeptide coupling on DMPC-based liposomes
Combining DMPC with one or more lipids selected from DMPG and DMTAP and at least one sterol derivative to provide a lipid mixture; preparing liposomes from said lipid mixture; and covalently coupling one or more immunogenic polypeptides to said at least one sterol derivative such that the immunogenic polypeptide(s) or carbohydrate(s) are covalently linked to between 1% and 100% of said at least one sterol derivative.
Optional immunostimulatory components in the liposomes
The method uses liposomes that further include at least one additional component selected from monophosphoryl lipid A (MPL), resiquimod, flagellin, CpG, and α-galactosylceramide.
Covalent linkage to lysine residues
Immunogenic polypeptides covalently linked to sterol derivatives via lysine residues on the immunogenic polypeptides.
Quantified covalent linkage fraction on sterol derivatives
One or more immunogenic polypeptides are covalently linked to between about 5% and about 10% of at least one sterol derivative.
(Alkylene oxide)n moiety linkage length constraint
Forming the covalent linkage to the immunogenic polypeptide using an (alkylene oxide)n moiety with an average length n between 40 and 1000.
Processing to form liposomes from a lipid mixture
Preparing liposomes from said lipid mixture by drying the mixture, hydrating the dried mixture in an aqueous vehicle, and then sonication, extrusion, or homogenization of the hydrated mixture to form liposomes.
Across the independent claim and its dependents, the inventive concept centers on DMPC-based liposomes containing DMPG and/or DMTAP plus a sterol-derivative covalent anchor, followed by covalent coupling of immunogenic polypeptides or carbohydrates to a defined fraction of the sterol derivative, with optional immunostimulatory components and defined linkage attachment and chemistry refinements.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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