Stabilized protein crystals, formulations comprising 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 or nucleic acid crystals, formulations and compositions comprising them. Methods are provided for the crystallization of proteins and nucleic acids and for the preparation of stabilized protein or nucleic acid crystals for use in dry or slurry formulations. The present invention is further directed to encapsulating proteins, glycoproteins, enzymes, antibodies, hormones and peptide crystals or crystal formulations into compositions for biological delivery to humans and animals. According to this invention, protein crystals or crystal formulations are encapsulated within a matrix comprising a polymeric carrier to form a composition. The formulations and compositions enhance preservation of the native biologically active tertiary structure of the proteins and create a reservoir which can slowly release active protein where and when it is needed. Methods are provided preparing stabilized formulations using pharmaceutical ingredients or excipients and optionally encapsulating them in a polymeric carrier to produce compositions and using such protein crystal formulations and compositions for biomedical applications, including delivery of therapeutic proteins and vaccines. Additional uses for the protein crystal formulations and compositions of this invention involve protein delivery in human food, agricultural feeds, veterinary compositions, diagnostics, cosmetics and personal care compositions.

1 - 32 . (canceled) 
     
     
         33 . A composition for the release of a protein, said composition comprising:
 (a) a protein crystal formulation, wherein said formulation comprises a protein crystal and an excipient; 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 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 T1/2.   
     
     
         34 - 35 . (canceled) 
     
     
         36 . A composition for the release of a protein, said composition comprising:
 (a) a protein crystal formulation, wherein said formulation comprises a protein crystal and an excipient; 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 the loss of less than 20% of α-helical structural content of the protein after storage for 4 days at 50° wherein the soluble form of said protein loses more than 50% of its α-helical structural content after storage for 6 hours at 50° C., as measured by FTIR.   
     
     
         37 . (canceled) 
     
     
         38 . A composition for the release of proteins, said composition comprising:
 (a) a protein crystal formulation, wherein said protein is greater than about 10,000 Daltons in molecular weight; and   (b) at least one polymeric carrier, wherein said formulation is encapsulated within a matrix of said polymeric carrier   
     
     
         39 - 77 . (canceled) 
     
     
         78 . A protein crystal composition comprising:
 (a) at least one polymeric carrier; and   (b) protein crystals, wherein said protein crystals are encapsulated within a matrix of said polymeric carrier.   
     
     
         79 . The protein crystal composition according to  claim 78 , wherein said protein is selected from the group consisting of glycoproteins, enzymes, hormones, antibodies, cytokines peptides. 
     
     
         80 - 187 . (canceled)