Stabilized Protein Crystals, Formulations Containing Them and Methods of Making Them

Abstract

This invention relates to methods for the stabilization, storage and delivery of biologically active macromolecules, such as proteins, peptides and nucleic acids. In particular, this invention relates to protein crystals, formulations and compositions comprising them. Provided are methods and compositions for encapsulating proteins, glycoproteins, enzymes, antibodies, hormones and peptide crystals or crystal formulations into compositions for biological delivery.

1 . A formulation, said formulation comprising:
 (a) a protein crystal, and   (b) at least one polymeric carrier, wherein said formulation is encapsulated within a matrix of said polymeric carrier such that the formulation is characterized by at least a 60 fold greater shelf life when stored at 50° C. than the soluble form of said protein in solution at 50° C., as measured by T 1/2 .   
     
     
         2 .- 10 . (canceled) 
     
     
         11 . The formulation according to  claim 1 , wherein said protein is a therapeutic protein. 
     
     
         12 . The formulation according to  claim 11 , wherein said therapeutic protein is an antibody. 
     
     
         13 . The formulation according to  claim 11 , wherein said therapeutic protein is a vaccine antigen. 
     
     
         14 . (canceled) 
     
     
         15 . The formulation according to  claim 1 , wherein said ingredient is further comprising an excipient. 
     
     
         16 . The formulation according to  claim 15 , wherein said excipient is selected from the group consisting of sucrose, trehalose, lactitol, gelatin, hydroxypropyl-β-cyclodextrin, methoxypolyethylene glycol and polyethylene glycol. 
     
     
         17 .- 30 . (canceled) 
     
     
         31 . A method of treating a mammal comprising the step of administering to the mammal an effective amount of the formulation according to  claim 1 . 
     
     
         32 . (canceled) 
     
     
         33 . (canceled) 
     
     
         34 . A composition for the release of a protein, said composition comprising:
 (a) a protein crystal formulation, and   (b) at least one polymeric carrier, wherein said formulation is encapsulated within a matrix of said polymeric carrier, and wherein said formulation is characterized by at least a 59 fold greater shelf life when stored at 40° C. and 75% humidity than the nonformulated form of said protein crystal when stored at 40° C. and 75% humidity, as measured by T 1/2 .   
     
     
         35 .- 41 . (canceled) 
     
     
         42 . The composition according to  claim 34 , wherein said protein is a therapeutic protein. 
     
     
         43 . The composition according to  claim 42 , wherein said therapeutic protein is selected from the group consisting of antibodies and, human growth hormones. 
     
     
         44 . The composition according to  claim 42 , wherein said therapeutic protein is a vaccine antigen. 
     
     
         45 .- 47 . (canceled) 
     
     
         48 . The composition according to  claim 34 , wherein said polymeric carrier is a biodegradable polymer. 
     
     
         49 . The composition according to  claim 34  wherein said polymeric carrier is a biocompatible polymer. 
     
     
         50 . The composition according to  claim 34  wherein said polymeric carrier is a polymer selected from one or more of the group consisting of poly (acrylic acid), poly (cyanoacrylates), poly (amino acids), poly (anhydrides), poly (depsipeptide), poly (esters), poly (lactic acid), poly (lactic-co-glycolic acid) or PLGA, poly (β-hydroxybutryate), poly (caprolactone), poly (dioxanone); poly (ethylene glycol), poly ((hydroxypropyl)methacrylamide, poly [(organo)phosphazene], poly (ortho esters), poly (vinyl alcohol), poly (vinylpyrrolidone), maleic anhydride-alkyl vinyl ether copolymers, pluronic polyols, albumin, alginate, cellulose and cellulose derivatives, collagen, fibrin, gelatin, hyaluronic acid, oligosaccharides, glycaminoglycans, sulfated polysaccharides, blends and copolymers thereof. 
     
     
         51 .- 109 . (canceled) 
     
     
         110 . A process for producing microspheres by encapsulating protein crystals, comprising the steps of:
 (a) suspending said protein crystals in a polymeric carrier dissolved in an organic solvent to produce a suspension of coated protein crystals;   (b) transferring said suspension of coated protein crystals, polymeric carrier and organic solvent to an aqueous solution containing an emulsifier; and   (c) hardening said polymeric carrier by evaporating the organic solvent in the presence of said emulsifier to produce said microspheres.   
     
     
         111 . The process according to  claim 110 , wherein said protein is selected from the group consisting of glycoproteins, enzymes, hormones, antibodies and peptides. 
     
     
         112 . The process according to  claim 110 , wherein said polymeric carrier is a biodegradable polymer. 
     
     
         113 . The process according to  claim 110 , wherein said polymeric carrier is a biocompatible polymer. 
     
     
         114 . The process according to  claim 110 , wherein said emulsifier is polyvinyl alcohol. 
     
     
         115 . The process according to  claim 110 , wherein said emulsifier is a surfactant. 
     
     
         116 . The process according to  claim 110 , wherein said polymeric carrier is a polymer selected from one or more of the group consisting of poly (acrylic acid), poly (cyanoacrylates), poly (amino acids), poly (anhydrides), poly (depsipeptide), poly (esters), poly (lactic acid), poly (lactic-co-glycolic acid) or PLGA, poly (β-hydroxybutryate), poly (caprolactone), poly (dioxanone), poly (ethylene glycol), poly ((hydroxypropyl)methacrylamide, poly [(organo)phosphazene], poly (ortho esters), poly (vinyl alcohol), poly (vinylpyrrolidone), maleic anhydride-alkyl vinyl ether copolymers, pluronic polyols, albumin, alginate, cellulose and cellulose derivatives, collagen, fibrin, gelatin, hyaluronic acid, oligosaccharides, glycaminoglycans, sulfated polysaccharides, blends and copolymers thereof. 
     
     
         117 .- 187 . (canceled)