US2009075890A1PendingUtilityA1

Long lasting synthetic exendin-4 peptide conjugates and methods of use thereof

Assignee: CONJUCHEM BIOTECHNOLOGIES INCPriority: May 17, 1999Filed: Aug 1, 2008Published: Mar 19, 2009
Est. expiryMay 17, 2019(expired)· nominal 20-yr term from priority
A61P 3/10A61K 47/643C07K 14/005C07K 14/57563A61K 38/00C07K 14/605A61P 1/00C12N 2740/16122
67
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Claims

Abstract

Modified insulinotropic peptides are disclosed. The modified insulinotropic peptides are capable of forming a peptidase stabilized insulinotropic peptide. The modified insulinotropic peptides are capable of forming covalent bonds with one or more blood components to form a conjugate. The conjugates may be formed in vivo or ex vivo. The modified peptides are administered to treat humans with diabetes and other related diseases.

Claims

exact text as granted — not AI-modified
1 - 19 . (canceled) 
     
     
         20 . A modified peptide comprising:
 a peptide; and   a maleimide or a maleimido-containing reactive group coupled, optionally via a linker, to the peptide,   the modified peptide being capable of covalently binding in vivo or in vitro by reaction of the reactive group, with a functionality on a blood component;   wherein the peptide comprises exendin-4 or a C-terminal amide thereof.   
     
     
         21 . The modified peptide of  claim 20 , wherein the amino acid sequence of the peptide is that of exendin-4(1-39) or a C-terminal amide thereof. 
     
     
         22 . The modified peptide of  claim 20 , wherein the amino acid sequence of the peptide is that of exendin-4(1-39) Lys 40  or a C-terminal amide thereof. 
     
     
         23 . The modified peptide of  claim 22  which is according to the following formula: 
       
         
           
           
               
               
           
         
       
     
     
         24 . The modified peptide of any one of  claims 20 - 23 , wherein the blood component is albumin. 
     
     
         25 . The modified peptide of  claim 20 , wherein the functionality on the blood component is an amino group, an hydroxyl group, or a thiol group. 
     
     
         26 . A conjugate formed by conjugating
 (a) a peptide coupled, optionally via a linker, to a maleimide or a maleimido-containing reactive group, with   (b) a blood component, by covalent linking of the reactive group with a functionality on the blood component,   wherein the peptide comprises exendin-4 or a C-terminal amide thereof.   
     
     
         27 . The conjugate of  claim 26 , wherein the amino acid sequence of the peptide is that of exendin-4(1-39) or a C-terminal amide thereof. 
     
     
         28 . The conjugate of  claim 26 , wherein the amino acid sequence of the peptide is that of exendin-4(1-39) Lys 40  or a C-terminal amide thereof. 
     
     
         29 . The conjugate of  claim 26  which is according to the formula: 
       
         
           
           
               
               
           
         
         wherein X is S, O, or NH of an amino acid of the blood component; 
         wherein L is present or absent, and when present is selected from the group consisting of —NH(CH 2 ) 2 NH—, —[C(O)—CH 2 —O—(CH 2 ) 2 —NH] m —, and —[C(O)—CH 2 —O—(CH 2 ) 2 —O—(CH 2 ) 2 —NH] m —; 
         wherein Z is a peptide comprising exendin-4 or a C-terminal amide thereof; and 
         wherein m is 1, 2 or 3. 
       
     
     
         30 . A conjugate according to the formula: 
       
         
           
           
               
               
           
         
         wherein X is S, O, or NH of an amino acid of the blood component; 
         wherein L is present or absent, and when present is selected from the group consisting of —NH(CH 2 ) 2 NH—, —[C(O)CH 2 —O—(CH 2 ) 2 —NH] m —, and —[C(O)—CH 2 —O—(CH 2 ) 2 —O—(CH 2 ) 2 —NH] m —; 
         wherein Z is a peptide comprising exendin-4 or a C-terminal amide thereof; and 
         wherein m is 1, 2 or 3. 
       
     
     
         31 . The conjugate of  claim 30 , wherein the amino acid sequence of the peptide is that of exendin-4(1-39) or a C-terminal amide thereof. 
     
     
         32 . The conjugate of  claim 30 , wherein the amino acid sequence of the peptide is that of exendin-4(1-39) Lys 40  or a C-terminal amide thereof. 
     
     
         33 . The conjugate of  claim 30 , wherein L is present. 
     
     
         34 . The conjugate of  claim 33 , wherein Z is covalently attached to L through the NH of an amino acid of Z. 
     
     
         35 . The conjugate of  claim 33 , wherein L is a poly ethoxy amino acid. 
     
     
         36 . The conjugate of  claim 35 , wherein the poly ethoxy amino acid is according to the formula:
   —[C(O)—CH 2 —O—(CH 2 ) 2 —NH] m —   wherein m is 1, 2 or 3.   
     
     
         37 . The conjugate of  claim 35 , wherein the poly ethoxy amino acid is according to the formula:
   —[C(O)—CH 2 —O—(CH 2 ) 2 —O—(CH 2 ) 2 —NH] m —   wherein m is 1, 2 or 3.   
     
     
         38 . The conjugate of  claim 30 , wherein X is S. 
     
     
         39 . The conjugate of  claim 30 , wherein the blood component is albumin. 
     
     
         40 . The conjugate of  claim 38 , wherein the S is of Cys-34 of human serum albumin. 
     
     
         41 . A conjugate according to the following formula: 
       
         
           
           
               
               
           
         
         wherein X is S of Cysteine 34 of albumin. 
       
     
     
         42 . The conjugate of  claim 41 , wherein said albumin is human serum albumin. 
     
     
         43 . A pharmaceutical composition comprising the modified peptide of any of  claims 20  to  25  and a physiologically acceptable medium. 
     
     
         44 . A pharmaceutical composition comprising the conjugate of any of  claims 26  to  42  and a physiologically acceptable medium. 
     
     
         45 . A method of synthesizing a modified exendin-4(1-39) peptide comprising:
 (a) synthesizing a peptide from the carboxy terminal amino acid, wherein the amino acid sequence of the peptide is that of exendin-4(1-39); and   (b) adding a reactive group, optionally via a linking group, to the carboxy terminal amino acid.   
     
     
         46 . A method of synthesizing a modified exendin-4(1-39) peptide comprising:
 (a) synthesizing a peptide from the carboxy terminal amino acid, wherein the amino acid sequence of the peptide is that of exendin-4(1-39) Lys 40 ; and   (b) adding a reactive group, optionally via a linking group, to the carboxy terminal amino acid.   
     
     
         47 . The method of  claim 45  or  46 , wherein the reactive group is maleimide or a maleimido-containing group. 
     
     
         48 . The method of  claim 45  or  46 , wherein the reactive group is succinimide or a succinimido-containing group. 
     
     
         49 . A method of preparing a conjugate comprising:
 contacting a modified peptide of  claim 20  with a blood component under conditions suitable for formation of a conjugate between said modified peptide and said blood component.   
     
     
         50 . The method of  claim 49 , wherein the amino acid sequence of the modified peptide is that of exendin-4(1-39) or a C-terminal amide thereof. 
     
     
         51 . The method of  claim 49 , wherein the amino acid sequence of the modified peptide is that of exendin-4(1-39) Lys 40  or a C-terminal amide thereof. 
     
     
         52 . The method of  claim 49 , wherein the reactive group of the modified peptide is a maleimido-containing group. 
     
     
         53 . The method of  claim 49 , wherein the blood component is albumin. 
     
     
         54 . The method of  claim 53 , wherein said modified peptide is conjugated to Cys-34 of human serum albumin. 
     
     
         55 . The method of  claim 49 , wherein the contacting occurs in vitro. 
     
     
         56 . The method of  claim 49 , wherein the modified peptide is according to the following formula: 
       
         
           
           
               
               
           
         
       
     
     
         57 . A method for treating diabetes in a subject with diabetes, comprising administering to the subject a therapeutically effective amount of the modified peptide of  claim 23 . 
     
     
         58 . The method of  claim 57 , wherein said modified compound is in association with a physiologically acceptable medium. 
     
     
         59 . A method for treating diabetes in a subject with diabetes, comprising administering to the subject a therapeutically effective amount of the conjugate of  claim 41 . 
     
     
         60 . The method of  claim 59 , wherein said albumin is human serum albumin. 
     
     
         61 . The method of  claim 59 , wherein said conjugate is in association with a physiologically acceptable medium. 
     
     
         62 . The method of  claim 60 , wherein said conjugate is in association with a physiologically acceptable medium. 
     
     
         63 . The method of any one of  claims 57  to  62 , wherein the diabetes is Type II diabetes. 
     
     
         64 . The method of any one of  claims 57  to  62 , wherein the subject is a human. 
     
     
         65 . A method for enhancing the expression of insulin in a subject, comprising administering to the subject a therapeutically effective amount of the modified peptide of  claim 23 . 
     
     
         66 . The method of  claim 65 , wherein said modified peptide is in association with a physiologically acceptable medium. 
     
     
         67 . A method for enhancing the expression of insulin in a subject, comprising administering to the subject a therapeutically effective amount of the conjugate of  claim 41 . 
     
     
         68 . The method of  claim 67 , wherein said albumin is human serum albumin. 
     
     
         69 . The method of  claim 67 , wherein said conjugate is in association with a physiologically acceptable medium. 
     
     
         70 . The method of  claim 68 , wherein said conjugate is in association with a physiologically acceptable medium. 
     
     
         71 . The method of any one of  claims 65  to  70 , wherein the subject is a human. 
     
     
         72 . A method for treating obesity in an obese subject, comprising administering to the subject a therapeutically effective amount of the modified peptide of  claim 23 . 
     
     
         73 . The method of  claim 72 , wherein said modified peptide is in association with a physiologically acceptable medium. 
     
     
         74 . A method for treating obesity in an obese subject, comprising administering to the subject a therapeutically effective amount of the conjugate of  claim 41 . 
     
     
         75 . The method of  claim 74 , wherein said albumin is human serum albumin. 
     
     
         76 . The method of  claim 74 , wherein said conjugate is in association with a physiologically acceptable medium. 
     
     
         77 . The method of  claim 75 , wherein said conjugate is in association with a physiologically acceptable medium. 
     
     
         78 . The method of any one of  claims 72  to  77 , wherein the subject is a human. 
     
     
         79 . A method for treating decreased insulin sensitivity in a subject with decreased insulin sensitivity, comprising administering to the subject a therapeutically effective amount of the modified peptide of  claim 23 . 
     
     
         80 . The method of  claim 79 , wherein said modified peptide is in association with a physiologically acceptable medium. 
     
     
         81 . A method for treating decreased insulin sensitivity in a subject with decreased insulin sensitivity, comprising administering to the subject a therapeutically effective amount of the conjugate of  claim 41 . 
     
     
         82 . The method of  claim 81 , wherein said albumin is human serum albumin. 
     
     
         83 . The method of  claim 81 , wherein said conjugate is in association with a physiologically acceptable medium. 
     
     
         84 . The method of  claim 82 , wherein said conjugate is in association with a physiologically acceptable medium. 
     
     
         85 . The method of any one of  claims 79  to  84 , wherein the subject is a human.

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