Gastrin compositions and formulations, and methods of use and preparation

Abstract

An embodiment of the invention provided herein is a pharmaceutical composition comprising a gastrin compound having an extended activity upon administration to a subject in comparison with native gastrin. Methods are provided of conjugating portions of the amino acid sequence of gastrin having functional ability to bind to the gastrin/CCK receptor, to various carrier moieties, including the use of amino acid spacer regions, and use of bifunctional cross-linking reagents. Methods of treating a diabetes patient with the compositions are provided.

What is claimed is:  
     
         1 . A pharmaceutical composition comprising a gastrin compound having an extended activity upon administration to a subject in comparison with native gastrin.  
     
     
         2 . A gastrin compound comprising: Z-Y m -X n -AA 1 -AA 2 -AA 3 -AA 4 -AA 5 -AA 6 , wherein AA 1 , is Tyr or Phe, AA 2  is Gly, Ala, or Ser, AA 3  is Trp, Val, or Ile, AA 4  is Met or Leu, AA 5  is Asp or Glu, and AA 6  is Phe or Tyr the AA 6  being amidated; wherein Z is a polymer which when the polymer is a protein, Z is the amino acid sequence of the protein; Y m  is an optional spacer region comprising m amino acid residues of a small neutral amino acid, and X is selected from any consecutive portions of: residues 1-28 of SEQ ID NO: 1, residues 1-28 of SEQ ID NO: 2, residues 1-11 of SEQ ID NO: 3, and residues 1-11 of SEQ ID NO: 4, providing that the gastrin compound binds a gastrin/CCK receptor.  
     
     
         3 . The gastrin compound according to  claim 2 , wherein AA 1 -AA 2 -AA 3 -AA 4 -AA 5 -AA 6  is Tyr-Gly-Trp-Met-Asp-Phe.  
     
     
         4 . The gastrin compound according to  claim 2 , wherein AA 1 -AA 2 -AA 3 -AA 4 -AA 5 -AA 6  is Tyr-Gly-Trp-Leu-Asp-Phe.  
     
     
         5 . The gastrin compound according to  claim 2 , wherein Z is a protein.  
     
     
         6 . The gastrin compound according to  claim 5 , wherein Z is human serum albumin.  
     
     
         7 . The gastrin compound according to  claim 2 , wherein Y is a sequence comprising m residues having glycine alternating with alanine or having a random sequence of glycine and alanine.  
     
     
         8 . The gastrin compound according to  claim 2 , wherein X is selected from the group of sequences: position 1 to position 11 of SEQ ID NO: 3; position 1 to position 11 of SEQ ID NO: 4; position 2 to position 11 of SEQ ID NO: 3; and position 2 to position 11 of SEQ ID NO: 4.  
     
     
         9 . The gastrin compound according to  claim 2 , further comprising a cysteine residue at the amino terminus of Y when m is greater than 1, or at the amino terminus of X when m is 0.  
     
     
         10 . The gastrin compound according to  claim 2 , wherein m is 0 to about 20 residues.  
     
     
         11 . The gastrin compound according to  claim 2 , wherein X n -AA 1 -AA 2 -AA 3 -AA 4 -AA 5 -AA 6  further comprises a bifunctional cross-linking agent for linkage to Z if m is 0.  
     
     
         12 . The gastrin compound according to  claim 2  which is recombinantly produced.  
     
     
         13 . A nucleotide sequence encoding the gastrin compound according to  claim 2 .  
     
     
         14 . A cell carrying the nucleotide sequence according to  claim 13 .  
     
     
         15 . The cell according to  claim 14  which is a bacterial or a yeast cell.  
     
     
         16 . The bacterial cell according to  claim 15  which is selected from the group consisting of an  Escherichia,  a  Bacillus,  and a  Streptomyces.    
     
     
         17 . The yeast cell according to  claim 15  which is selected from the group consisting of a  Saccharomyces,  a  Kluyveromyces,  a  Schizosaccharomyces  and a  Pichia.    
     
     
         18 . The gastrin compound according to  claim 1  wherein the gastrin component contains at least amino acids selected from the group of: positions 29-34 of SEQ ID NO:1; positions 29-34 of SEQ ID NO:2; positions 12-17 of SEQ ID NO: 3; and positions 12-17 of SEQ ID NO: 4, and the gastrin is further associated with a protein, a polymer, a lipid or a carbohydrate.  
     
     
         19 . The gastrin compound according to either of claims  2  or  18 , wherein the gastrin component contains at least amino acids at positions 29-34 of SEQ ID NO:2 or positions 12-17 of SEQ ID NO:4.  
     
     
         20 . The gastrin compound according to  claim 18 , wherein the polymer is a polyethylene glycol (PEG) or a dextran.  
     
     
         21 . The gastrin compound according to  claim 18 , wherein the protein is a serum albumin.  
     
     
         22 . The gastrin compound according to  claim 21 , wherein the serum albumin is human serum albumin.  
     
     
         23 . A gastrin compound comprising a structure C-Y m -X, wherein C is Cys or Lys, Y m  is an optional spacer region comprising m amino acid residues of a small neutral amino acid, and X is at least six amino acid residues comprising sequences selected from at least positions 12-17 of gastrin-17 (SEQ ID NO: 3 and 4) and at least positions 29-34 of gastrin-34 (SEQ ID NO: 1 and 2).  
     
     
         24 . The gastrin compound according to  claim 23 , further conjugated to a polymer.  
     
     
         25 . The gastrin compound according to  claim 23 , further conjugated to a polyethylene glycol (PEG) or a dextran.  
     
     
         26 . The gastrin compound according to  claim 23 , further conjugated to a protein.  
     
     
         27 . The gastrin compound according to  claim 23 , further comprising a bifunctional cross-linking agent wherein a first reactive end of the cross-linking agent is covalently linked to C.  
     
     
         28 . The gastrin compound according to  claim 23 , wherein a second reactive end of the cross-linking agent is covalently linked to a polymer or protein.  
     
     
         29 . The gastrin compound according to  claim 23 , wherein C-Y m -X is produced recombinantly or is synthesized by peptide synthesis.  
     
     
         30 . The gastrin compound according to any of claims  1 ,  2  and  23 , in an effective dose.  
     
     
         31 . The gastrin compound according to any of claims  1 ,  2  and  23 , further comprising an agent for immune suppression.  
     
     
         32 . The gastrin compound according to any of claims  1 ,  2  and  23 , further comprising a growth factor.  
     
     
         33 . The gastrin compound according to  claim 32 , wherein the growth factor is a glucagon-like peptide 1 receptor ligand.  
     
     
         34 . The gastrin compound according to  claim 32 , wherein the growth factor is an EGF receptor ligand.  
     
     
         35 . The gastrin compound according to  claim 1 ,  2  and  23 , further comprising a hypoglycemic agent.  
     
     
         36 . The gastrin compound according to any of claims  1 ,  2  and  23 , further comprising a pharmaceutically acceptable carrier.  
     
     
         37 . A method of treating a subject having diabetes, comprising administering a gastrin compound according to any of claims  1 ,  2  and  23 .  
     
     
         38 . The method according to  claim 37 , wherein frequency of administering the gastrin compound is less than frequency of administration of a native gastrin.  
     
     
         39 . The method according to  claim 37 , further comprising measuring a physiological indicator of islet neogenesis.  
     
     
         40 . The method according to  claim 37 , further comprising measuring fasting blood glucose (FBG).  
     
     
         41 . The method according to  claim 37 , further comprising decreasing insulin dependency.  
     
     
         42 . A method of making a gastrin compound comprising associating an amino acid sequence of a gastrin with a carrier composition.  
     
     
         43 . The method according to  claim 42 , wherein prior to associating the gastrin with the carrier, the gastrin is modified to comprise a cysteine substitution or an additional cysteine residue.  
     
     
         44 . The method according to  claim 43 , wherein the cysteine substitution is a replacement of pyroglutamate.  
     
     
         45 . The method according to  claim 42 , wherein the gastrin amino acid sequence comprises at least positions selected from the group of: residues 29-34 of amino acid sequence SEQ ID NO: 1; residues 29-34 of amino acid sequence SEQ ID NO: 2; residues 12-17 of amino acid sequence SEQ ID NO: 3; and residues 12-17 of amino acid sequence SEQ ID NO: 4.  
     
     
         46 . The method according to  claim 43 , wherein the cysteine is at the amino terminus of the gastrin.  
     
     
         47 . The method according to  claim 42 , further comprising prior to associating the gastrin with the carrier, modifying the gastrin to further comprise a bifunctional cross-linking agent.  
     
     
         48 . A method of treating a diabetes patient comprising administering to the patient a modified gastrin capable of covalently reacting with a serum protein.  
     
     
         49 . The method according to  claim 48 , wherein the modified gastrin comprises a sequence of a native gastrin capable of binding to the gastrin/CCK receptor and an amino terminal cysteine or lysine.  
     
     
         50 . The method according to  claim 42 , wherein the sequence of the native gastrin is selected from the group of: residues 29-34 of amino acid sequence SEQ ID NO: 1; residues 29-34 of amino acid sequence SEQ ID NO: 2; residues 12-17 of amino acid sequence SEQ ID NO: 3; and residues 12-17 of amino acid sequence SEQ ID NO: 4.  
     
     
         51 . A method for maintaining for an extended period of time an increased gastrin serum level compared with the serum level of a peptide having an amino acid sequence of a gastrin, the method comprising administering a gastrin compound according to any of  claim 1 ,  2  and  23 .  
     
     
         52 . A kit comprising at least one effective dose of a gastrin compound according to any of claims  1 ,  2  and  23 .  
     
     
         53 . A gastrin compound according to  claim 9 , further comprising a bifunctional crosslinking agent for linkage to Z.