Glucagon-like peptide-1 crystals

Abstract

The invention provides individual tetragonal flat rod shaped or plate-like crystals of glucagon-like peptide-1 related molecules, processes for their preparation, compositions and methods of use. The crystal preparations exhibit extended time action in vivo and are useful for treating diabetes, obesity and related conditions.

1. Crystals comprising zinc, and a GLP-1 analog wherein the crystals have a tetragonal flat rod shaped or plate-like morphology and the GLP-1 analog is a peptide of the formula:
 R 1 -X-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Y-Gly-Gln-Ala-Ala-Lys-Z-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-R 2  SEQ ID NO:2  
 
       wherein: R 1  is selected from the group consisting of L-histidine, D-histidine, desamino-histidine, 2-amino-histidine, beta-hydroxy-histidine, homohistidine, alpha-fluoromethyl-histidine, and alpha-methyl-histidine; X is selected from the group consisting of Ala, Gly, Val, Thr, Met, Ile, and alpha-methyl-Ala; Y is selected from the group consisting of Glu, Gln, Ala, Thr, Ser, and Gly; Z is selected from the group consisting of Glu, Gln, Ala, Thr, Ser, and Gly; and R 2  is selected from the group consisting of NH 2 , and Gly-OH. 
     
     
       2. The crystals of  claim 1  wherein X is selected from the group consisting of Gly, Val, Thr, Met, Ile, and alpha-methyl-Ala. 
     
     
       3. The crystals of  claim 2  having a size from between about 2-25 microns by 10-150 microns with a depth from about 0.5-5 microns. 
     
     
       4. A pharmaceutical formulation comprising crystals as claimed in  claim 1  together with one or more pharmaceutical acceptable diluents, carriers, or excipients. 
     
     
       5. A pharmaceutical formulation comprising crystals as claimed in  claim 2  with one or more pharmaceutically acceptable diluents, carriers, or excipients. 
     
     
       6. A pharmaceutical formulation comprising crystals as claimed in  claim 3  with one or more pharmaceutically acceptable diluents, carriers, or excipients. 
     
     
       7. Crystals comprising zinc, and a GLP-   1  analog wherein the crystals have a tetragonal flat rod shaped or plate - like morphology and the GLP -   1  analog has at least one modification selected from the group consisting of:    ( a )  substitution of glycine, serine, cysteine, threonine, asparagine, glutamine, tyrosine, alanine, valine, isoleucine, leucine, methionine, phenylalanine, arginine, or D - lysine for lysine at position  26  and/or position  34 ; or substitution of glycine, serine, cysteine, threonine, asparagine, glutamine, tyrosine, alanine, valine, isoleucine, leucine, methionine, phenylalanine, lysine, or D - arginine for arginine at position  36 ;      ( b )  substitution of an oxidation - resistant amino acid for tryptophan at position  31 ;      ( c )  substitution of at least one of tyrosine for valine at position  16 ; lysine for serine at position  18 ; aspartic acid for glutamic acid at position  21 ; serine for glycine at position  22 ; arginine for glutamine at position  23 ; arginine for alanine at position  24 ; and glutamine for lysine at position  26 ; and      ( d )  substitution comprising at least one of: glycine, serine, or cysteine for alanine at position  8 ; aspartic acid, glycine, serine, cysteine, threonine, asparagine, glutamine, tyrosine, alanine, valine, isoleucine, leucine, methionine, or phenylalanine for glutamic acid at position  9 ; serine, cysteine, threonine, asparagine, glutamine, tyrosine, alanine, valine, isoleucine, leucine, methionine, or phenylalanine for glycine at position  10 ; and glutamic acid for aspartic acid at position  15 ; and      ( e )  substitution of glycine, serine, cysteine, threonine, asparagine, glutamine, tyrosine, alanine, valine, isoleucine, leucine, methionine, or phenylalanine or the D - or N - acylated or alkylated form of histidine for histidine at position  7 .     
     
     
       8. The crystals of  claim 7 , wherein the GLP-   1  analog comprises an arginine substituted for lysine at position  34 .   
     
     
       9. The crystals of  claim 8 , wherein an epsilon- amino group of lysine is acylated.   
     
     
       10. The crystals of  claim 7  having a size from between about  2 -   25  microns by  10   -   150  microns with a depth from about  0 . 5   -   5  microns.   
     
     
       11. The crystals of  claim 8  having a size from between about  2 -   25  microns by  10   -   150  microns with a depth from about  0 . 5   -   5  microns.   
     
     
       12. The crystals of  claim 9  having a size from between about  2 -   25  microns by  10   -   150  microns with a depth from about  0 . 5   -   5  microns.   
     
     
       13. A pharmaceutical formulation comprising crystals as claimed in  claim 8  with one or more pharmaceutically acceptable diluents, carriers, or excipients. 
     
     
       14. A pharmaceutical formulation comprising crystals as claimed in  claim 9  with one or more pharmaceutically acceptable diluents, carriers, or excipients.