Urocortin 2 analogs and uses thereof

Abstract

Disclosed are polypeptides that are analogs of urocortin 2 that have pharmacological activity similar to urocortin 2 but have improved water solubility compared to urocortin 2, and pharmaceutical compositions of the polypeptides of the present invention. Also disclosed are polynucleotides encoding the polypeptides, and methods of treating pathophysiological states employing pharmaceutical compositions of the polypeptides and polynucleotides of the present invention. In addition, disclosed are vectors and host cells that include a nucleic acid encoding a polypeptide of the present invention, and kits that include pharmaceutical compositions of the present invention.

1 . An isolated polypeptide of formula (I):
   X 1 —X 2 —X 3 —X 4 —X 5 —X 6   (I)
   wherein X 1  is either absent or a polypeptide having 1-200 residues;   X 2  is absent or 1-20 amino acid residues that are each individually selected from either arginine (R) or lysine (K);   X 3  is SEQ ID NO:1, SEQ ID NO:19, SEQ ID NO:21, or SEQ ID NO:22;   X 4  is either absent, G, GH, or GHC;   X 5  is absent or 1-20 amino acid residues that are each individually selected from either arginine (R) or lysine (K);   X 6  is either absent or a polypeptide having 1-200 residues;   provided that if X 3  is SEQ ID NO:1, X 4 , X 5 , and X 6  are all absent, then X 2  is not absent; further provided that if X 3  is SEQ ID NO:19, X 5  is absent, and X 6  is absent, then X 4  is G, GH, or GHC.   
     
     
         2 . The isolated polypeptide of  claim 1 , wherein X 3  is SEQ ID NO:1. 
     
     
         3 . The isolated polypeptide of  claim 1 , wherein X 3  is SEQ ID NO:19. 
     
     
         4 . The isolated polypeptide of  claim 2 , wherein X 2  is arginine (R). 
     
     
         5 . The isolated polypeptide of  claim 1 , wherein the polypeptide is further defined as being amidated at the C-terminus. 
     
     
         6 . The isolated polypeptide of  claim 5 , wherein the polypeptide is SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:9. 
     
     
         7 . The isolated polypeptide of  claim 6 , wherein the polypeptide is SEQ ID NO:5. 
     
     
         8 . The isolated polypeptide of  claim 1 , wherein X 2  is lysine (K). 
     
     
         9 . The isolated polypeptide of  claim 1 , wherein the polypeptide comprises SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, or SEQ ID NO:18. 
     
     
         10 . A recombinant nucleic acid comprising a nucleic acid segment encoding for a polypeptide of  claim 1 . 
     
     
         11 . The recombinant nucleic acid of  claim 10 , wherein X 3  is SEQ ID NO:1. 
     
     
         12 . The recombinant nucleic acid of  claim 10 , wherein X 3  is SEQ ID NO:19. 
     
     
         13 . The recombinant nucleic acid of  claim 11 , wherein X 2  is arginine (R). 
     
     
         14 . The recombinant nucleic acid of  claim 10 , wherein X 2  is lysine (K). 
     
     
         15 . The recombinant nucleic acid of  claim 10 , wherein X 4  is G. 
     
     
         16 . The recombinant nucleic acid of  claim 10 , wherein X 4  is absent. 
     
     
         17 . The recombinant nucleic acid of  claim 10 , wherein the polypeptide is SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, or SEQ ID NO:18. 
     
     
         18 . A chimeric polypeptide comprising:
 a) a first amino acid sequence that is a polypeptide as set forth in  claim 1 ; and   b) a second amino acid sequence, wherein the second amino acid sequence is a therapeutic amino acid sequence.   
     
     
         19 . The chimeric polypeptide of  claim 18 , wherein the second amino acid sequence enhances the bioavailability of the first amino acid sequence. 
     
     
         20 . The chimeric polypeptide of  claim 19 , wherein the second amino acid sequence is selected from the group consisting of a low density lipoprotein receptor binding domain of apolipoprotein B (SEQ ID NO:20), an Fc amino acid sequence, or a toxin. 
     
     
         21 . The chimeric polypeptide of  claim 20 , wherein the second amino acid sequence is a toxin selected from the group consisting of gelonin, dodecandrin, tricosanthin, tricokirin, bryodin, mirabilis antiviral protein, barley ribosome-inactivating protein (BRIP), pokeweed antiviral protein (PAPs), saporin, luffin, momordin, ricin, abrin, diphtheria toxin A, pertussis toxin A subunit,  E. coli  enterotoxin toxin A subunit, cholera toxin (CTX) and  Pseudomonas  toxin c-terminal. 
     
     
         22 . A polynucleotide encoding a chimeric polypeptide as set forth in  claim 18 . 
     
     
         23 . A pharmaceutical composition comprising:
 a) the isolated polypeptide of  claim 1  or a recombinant nucleic acid that encodes such a polypeptide; and   b) a pharmaceutically acceptable carrier.   
     
     
         24 . A pharmaceutical composition comprising:
 a) a chimeric polypeptide of  claim 18  or a polynucleotide encoding such a polypeptide; and   b) a pharmaceutically acceptable carrier.   
     
     
         25 . The pharmaceutical composition of  claim 24 , wherein the pharmaceutically acceptable carrier comprises water or normal saline. 
     
     
         26 . A method of treating a pathophysiological state in a subject, comprising administering to the subject a pharmaceutical composition comprising;
 a) an isolated polypeptide as set forth in  claim 1  or a recombinant nucleic acid that encodes such a polypeptide; and   b) a pharmaceutically acceptable carrier.   
     
     
         27 . The method of  claim 26 , wherein the subject is a mammal. 
     
     
         28 . The method of  claim 27 , wherein the mammal is a human, a primate, a horse, a cow, a sheep, a pig, a dog, a cat, a rat, or a mouse. 
     
     
         29 . The method of  claim 28 , wherein the mammal is a human. 
     
     
         30 . The method of  claim 26 , wherein the pathophysiological state is selected from the group consisting of high body temperature, appetite dysfunction, congestive heart failure, stress, anxiety, and undesirably low levels of ACTH secretion. 
     
     
         31 . A vector comprising a nucleic acid sequence as set forth in  claim 10 , wherein said vector further comprises regulatory elements necessary for expression of said nucleic acid sequence in a cell. 
     
     
         32 . The vector of  claim 31 , further defined as an viral vector. 
     
     
         33 . The vector of  claim 32 , wherein said viral vector is an adenoviral vector, a lentiviral vector, or an adeno-associated viral vector. 
     
     
         34 . A host cell transfected with the vector of  claim 31 , wherein said cell is selected from the group consisting of a bacterial cell, a mammalian cell, a plant cell, or an insect cell. 
     
     
         35 . A kit comprising a sealed container comprising a pharmaceutically acceptable composition as set forth in  claim 23 . 
     
     
         36 . The kit of  claim 35 , further comprising a secondary pharmaceutical agent that can be applied in the treatment of a pathophysiological state.