Contulakin-G, analogs thereof and uses therefor

Abstract

The present invention is directed to contulakin-G (which is the native glycosylated peptide), a des-glycosylated contulakin-G (termed Thr 10 -contulakin-G), and derivatives thereof, to a cDNA clone encoding a precursor of this mature peptide and to a precursor peptide. The invention is further directed to the use of this peptide as a therapeutic for anti-seizure, anti-inflammatory, anti-shock, anti-thrombus, hypotensive, analgesia, anti-psychotic, Parkinson's disease, gastrointestinal disorders, depressive states, cognitive dysfunction, anxiety, tardive dyskinesia, drug dependency, panic attack, mania, irritable bowel syndrome, diarrhea, ulcer, GI tumors, Tourette's syndrome, Huntington's chorea, vascular leakage, anti-arteriosclerosis, vascular and vasodilation disorders, as well as neurological, neuropharmalogical and neuropsychopharmacological disorders.

What is claimed is:  
     
         1 . A method for treating pain in an individual which comprises administering an analgesic effective amount of an active agent to an individual in need of pain treatment, said active agent comprises contulakin-G which comprises the amino acid sequence Xaa 1 -Ser-Glu-Glu-Gly-Gly-Ser-Asn-Ala-Thr-Lys-Lys-Xaa 2 -Tyr-Ile-Leu (SEQ ID NO:1), where Xaa 1  is pyro-Glu, Xaa 2  is proline or hydroxyproline and Thr 10  is modified to contain an O-glycan.  
     
     
         2 . The method of  claim 1 , wherein said pain is acute pain.  
     
     
         3 . The method of  claim 2 , wherein said acute pain is post-trauma.  
     
     
         4 . The method of  claim 1 , wherein said pain is chronic pain.  
     
     
         5 . The method of  claim 4 , wherein said chronic pain results from cancer.  
     
     
         6 . The method of  claim 4 , wherein said chronic pain is neuropathic pain.  
     
     
         7 . The method of  claim 4 , wherein said chronic pain is inflammatory.  
     
     
         8 . The method of  claim 1 , wherein the active agent is administered using a delivery system selected from the group consisting of infusion, pump delivery, bioerodable polymer delivery, microencapsulated cell delivery, injection and macroencapsulated cell delivery.  
     
     
         9 . The method of  claim 8 , wherein administration is into the central nervous system.  
     
     
         10 . The method of  claim 9 , wherein the central nervous system is selected from the group consisting of the intrathecal space, the brain ventricles and the brain parenchyma.  
     
     
         11 . The method of  claim 8 , wherein the administration is selected from the group consisting of subcutaneous, intravenous, intra-arterial and intramuscular.  
     
     
         12 . The method of  claim 1 , wherein the glycan is Gal(β1→3)GalNAc(α1→).  
     
     
         13 . The method of  claim 1 , wherein the glycan has the structure  
       
         
           
           
               
               
           
         
         wherein R 1  is Thr; X is 0; R 2  is OH, NH 2 , NHSO 3 Na, NHAc, O-sulphate, O-phosphate, or O-glycan; R 3  is H, SO 3 , PO 3 , acetyl, sialic acid or monosaccharide; R 4  is H, SO 3 , PO 3 , acetyl or monosaccharide; R 5  is OH, NH 2 , NHSO 3 Na, NHAc, O-sulphate, O-phosphate, O-monosaccharide or, O-acetyl; R 6  is H, SO 3 , PO 3 , acetyl or monosaccharide; R 7  is H, SO 3 , PO 3 , acetyl or monosaccharide; R 8  is H, SO 3 , PO 3 , acetyl or monosaccharide; n is 0-4 and m is 1-4.  
       
     
     
         14 . A method for treating pain in an individual which comprises administering an analgesic effective amount of an active agent to an individual in need of pain treatment, said active agent is selected from the group consisting of 
 (a) a generic contulakin-G having the following general formula Xaa 1 -Xaa 2 -Xaa 3 -Xaa 3 -Gly-Gly-Xaa 2 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa 8 -Xaa 9 -Xaa 10 -Ile-Leu (SEQ ID NO:2), where Xaa 1  is pyro-Glu, Glu, Gln or γ-carboxy-Glu; Xaa 2  is Ser, Thr or S-glycan modified Cys; Xaa 3  is Glu or γ-carboxy-Glu; Xaa 4  is Asn, N-glycan modified Asn or S-glycan modified Cys; Xaa 5  is Ala or Gly; Xaa 6  is Thr, Ser, S-glycan modified Cys, Tyr or any hydroxy containing unnatural amino acid; Xaa 7  is Lys, N-methyl-Lys, N,N-dimethyl-Lys, N,N,N-trimethyl-Lys, Arg, ornithine, homoarginine or any unnatural basic amino acid; Xaa 8  is Ala, Gly, Lys, N-methyl-Lys, N,N-dimethyl-Lys, N,N,N-trimethyl-Lys, Arg, ornithine, homoarginine, any unnatural basic amino acid or X-Lys where X is (CH 2 ) n , phenyl, —(CH 2 ) m —(CH═CH)—(CH 2 ) m H or —(CH 2 ) m —(C≡C)—(CH 2 ) m H in which n is 1-4 and m is 0-2; Xaa 9  is Pro or hydroxy-Pro; and Xaa 10  is Tyr, mono-iodo-Tyr, di-iodo-Tyr, O-sulpho-Tyr, O-phospho-Tyr, nitro-Tyr, Trp, D-Trp, bromo-Trp, bromo-D-Trp, chloro-Trp, chloro-D-Trp, Phe, L-neo-Trp, or any unnatural aromatic amino acid, with the proviso that the generic contulakin-G is not un-glycosylated contulakin-G;    (b) a generic contulakin-G of (a) which is modified to contain an O-glycan, an S-glycan or an N-glycan;    (c) a contulakin-G analog which comprises an N-terminal truncation of from 1 to 9 amino acids of the generic contulakin-G of (a);    (d) a contulakin-G analog of (c), wherein a Ser-O-glycan, Thr-O-glycan or Cys-S-glycan is substituted for the amino acid residue at the truncated N-terminus;    (e) a contulakin-G analog, wherein a Ser-O-glycan, Thr-O-glycan or Cys-S-glycan is substituted for a residue at positions 1-9 of the generic contulakin-G of (a); and    (f) a contulakin-G analog which comprises an N-terminal truncation of 10 amino acids of the generic contulakin-G of (a) which is further modified to contain a Lys-N-glycan at residue 11 of the generic contulakin-G.    
     
     
         15 . The method of  claim 14 , wherein said pain is acute pain.  
     
     
         16 . The method of  claim 15 , wherein said acute pain is post-trauma.  
     
     
         17 . The method of  claim 14 , wherein said pain is chronic pain.  
     
     
         18 . The method of  claim 17 , wherein said chronic pain results from cancer.  
     
     
         19 . The method of  claim 17 , wherein said chronic pain is neuropathic pain.  
     
     
         20 . The method of  claim 17 , wherein said chronic pain is inflammatory.  
     
     
         21 . The method of  claim 14 , wherein the active agent is administered using a delivery system selected from the group consisting of infusion, pump delivery, bioerodable polymer delivery, microencapsulated cell delivery, injection and macroencapsulated cell delivery.  
     
     
         22 . The method of  claim 21 , wherein administration is into the central nervous system.  
     
     
         23 . The method of  claim 22 , wherein the central nervous system is selected from the group consisting of the intrathecal space, the brain ventricles and the brain parenchyma.  
     
     
         24 . The method of  claim 21 , wherein the administration is selected from the group consisting of subcutaneous, intravenous, intra-arterial and intramuscular.  
     
     
         25 . The method of  claim 14 , wherein the glycan is Gal(β1→3)GalNAc(α1→).  
     
     
         26 . The method of  claim 14 , wherein the glycan has the structure  
       
         
           
           
               
               
           
         
         wherein R 1  is Thr, Ser, Cys, Asn or Lys; X is 0 when R is Thr or Ser, or X is S when R 1  is Cys or X is N when R 1  is Asn or Lys; R 2  is OH, NH 2 , NHSO 3 Na, NHAc, O-sulphate, O-phosphate, or O-glycan; R 3  is H, SO 3 , PO 3 , acetyl, sialic acid or monosaccharide; R 4  is H, SO 3 , PO 3 , acetyl or monosaccharide; R 5  is OH, NH 2 , NHSO 3 Na, NHAc, O-sulphate, O-phosphate, O-monosaccharide or, O-acetyl; R 6  is H, SO 3 , PO 3 , acetyl or monosaccharide; R 7  is H, SO 3 , PO 3 , acetyl or monosaccharide; R 8  is H, SO 3 , PO 3 , acetyl or monosaccharide; n is 0-4 and m is 1-4.