Non-Natural Amino Acids

Abstract

This invention relates to non-natural desamino alkyl amino acid compounds, methods of making, and peptides containing these compounds as their N-terminus moieties. A preferred example is neurotensin (8-13) in which the N terminus is an alpha desamino, alpha methyl N,N dimethyl homolysine residue.

1 . A non-natural desamino, alkyl amino acid compound having Formula I: 
       
         
           
           
               
               
           
         
         wherein 
         n is an integer of from 0 to 5; 
         m is zero or an integer of 1; 
         R is a straight or branched chain alkyl group of C 1 -C 6 , or an aromatic group of C 6 -C 18  or a corresponding substituted aromatic group with one or two substituents selected from halogen, alkyloxy, carboxy, amide or alkyl in any combination, or a heteroaromatic group of C 4 -C 18  and one or two heteroatoms selected from oxygen, sulfur and nitrogen in any combination or a corresponding substituted heteraromatic group with one or two substituents selected from halogen, alkyloxy, carboxy, amide or alkyl in any combination; 
         R 1 , R 2 , and R 3  are, independently, hydrogen or branched or straight chain alkyl, alkenyl or alkynyl of C 1 -C 6  or an aromatic group of C 6 -C 18  or a corresponding substituted aromatic group with one or two substituents selected from halogen, alkyloxy, carboxy, amide or alkyl in any combination, or a heteroaromatic group of C 4 -C 18  and one or two heteroatoms selected from oxygen, sulfur and nitrogen in any combination or a corresponding substituted heteraromatic group with one or two substituents selected from halogen, alkyloxy, carboxy, amide or alkyl in any combination and with the proviso that a maximum of two of R 1 , R 2 , and R 3  may be selected to be the aromatic, substituted aromatic, heteroaromatic or substituted heteroaromatic group; 
         and Cα is a carbon atom having either R or S stereochemistry; 
         or an ester, amide, alkyl amide or metal cation or ammonium salt of the carboxylic acid group thereof, or an organic or inorganic acid salt of the amine group thereof, or any combination thereof. 
       
     
     
         2 . The compound of  claim 1 , wherein the stereochemistry at Cα is S. 
     
     
         3 . The compound of  claim 1 , wherein R, R 1 , R 2 , and R 3  are, independently, hydrogen or methyl. 
     
     
         4 . The compound of  claim 1 , wherein n is an integer from 2 to 5. 
     
     
         5 . A non-natural desamino, alkyl amino acid compound of the formula II: 
       
         
           
           
               
               
           
         
         wherein 
         n is an integer of from 0 to 6; 
         when dashed line a is not present, X and Y are independently, hydrogen or lower branched or straight chain alkyl, alkenyl or alkynyl of C 1 -C 6 ; 
         when dashed line a is present, X—Y is (CH 2 ) z , wherein z is an integer of from 1-8; 
         R is a straight or branched chain alkyl group of C 1 -C 6 , or an aromatic group of C 6 -C 18  or a corresponding substituted aromatic group with one or two substituents selected from halogen, alkyloxy, carboxy, amide or alkyl in any combination, or a heteroaromatic group of C 4 -C 18  and one or two heteroatoms selected from oxygen, sulfur and nitrogen in any combination or a corresponding substituted heteraromatic group with one or two substituents selected from halogen, alkyloxy, carboxy, amide or alkyl in any combination; 
         R 4  is hydrogen or lower branched or straight chain alkyl, alkenyl or alkynyl of C 1 -C 6 , or an aromatic group of C 6 -C 18  or a corresponding substituted aromatic group with one or two substituents selected from halogen, alkyloxy, carboxy, amide or alkyl in any combination, or a heteroaromatic group of C 4 -C 18  and one or two heteroatoms selected from oxygen, sulfur and nitrogen in any combination or a corresponding substituted heteraromatic group with one or two substituents selected from halogen, alkyloxy, carboxy, amide or alkyl in any combination, and; 
         C α  is a carbon atom and the stereochemistry at C α  is either R or S; 
         or an ester, amide, alkyl amide or metal cation or ammonium salt of the carboxylic acid group thereof, or an organic or inorganic acid salt of the amine group thereof, or any combination thereof. 
       
     
     
         6 . The compound of  claim 5 , wherein n is an integer from 2 to 5 and
 z is an integer from 2 to 4.   
     
     
         7 . A non-natural desamino, alkyl amino acid compound of the formula III: 
       
         
           
           
               
               
           
         
         wherein 
         n is an integer of from 0 to 5; 
         X—Y is (CH 2 ) z , wherein z is an integer of from 0 to 6; 
         R is a straight or branched chain alkyl group of C 1 -C 6 , or an aromatic group of C 6 -C 18  or a corresponding substituted aromatic group with one or two substituents selected from halogen, alkyloxy, carboxy, amide or alkyl in any combination, or a heteroaromatic group of C 4 -C 18  and one or two heteroatoms selected from oxygen, sulfur and nitrogen in any combination or a corresponding substituted heteraromatic group with one or two substituents selected from halogen, alkyloxy, carboxy, amide or alkyl in any combination; 
         R 6 , and R 7  are, independently, hydrogen or lower branched or straight chain alkyl, alkenyl or alkynyl of C 1 -C 6 , or an aromatic group of C 6 -C 18  or a corresponding substituted aromatic group with one or two substituents selected from halogen, alkyloxy, carboxy, amide or alkyl in any combination, or a heteroaromatic group of C 4 -C 18  and one or two heteroatoms selected from oxygen, sulfur and nitrogen in any combination or a corresponding substituted heteraromatic group with one or two substituents selected from halogen, alkyloxy, carboxy, amide or alkyl in any combination; and 
         C α  is a carbon atom and the stereochemistry at C α  is either R or S; 
         or an ester, amide, alkyl amide or metal cation or ammonium salt of the carboxylic acid group thereof, or an organic or inorganic acid salt of the amine group thereof, or any combination thereof. 
       
     
     
         8 . The compound of  claim 7 , wherein n is an integer from 2 to 5 and z is an integer from 2 to 4. 
     
     
         9 . A non-natural desamino, alkyl amino acid compound of the formula IV: 
       
         
           
           
               
               
           
         
         wherein 
         n is an integer of from 0 to 5; 
         R is a straight or branched chain alkyl group of C 1 -C 6 , or an aromatic group of C 6 -C 18  or a corresponding substituted aromatic group with one or two substituents selected from halogen, alkyloxy, carboxy, amide or alkyl in any combination, or a heteroaromatic group of C 4 -C 18  and one or two heteroatoms selected from oxygen, sulfur and nitrogen in any combination or a corresponding substituted heteraromatic group with one or two substituents selected from halogen, alkyloxy, carboxy, amide or alkyl in any combination; 
         R 9 , R 10 , and R 11  are, independently, hydrogen or lower branched or straight chain alkyl, alkenyl or alkynyl of C 1 -C 6 , or an aromatic group of C 6 -C 18  or a corresponding substituted aromatic group with one or two substituents selected from halogen, alkyloxy, carboxy, amide or alkyl in any combination, or a heteroaromatic group of C 4 -C 18  and one or two heteroatoms selected from oxygen, sulfur and nitrogen in any combination or a corresponding substituted heteraromatic group with one or two substituents selected from halogen, alkyloxy, carboxy, amide or alkyl in any combination and with the proviso that a maximum of two of R 9 , R 10 , and R 11  may be selected to be the aromatic, substituted aromatic, heteroaromatic or substituted heteroaromatic group; and 
         C α  is a carbon atom and the stereochemistry at C α  is either R or S; 
         or an ester, amide, alkyl amide or metal cation or ammonium salt of the carboxylic acid group thereof, or an organic or inorganic acid salt of the amine group thereof, or any combination thereof. 
       
     
     
         10 . A compound of  claim 9 , wherein n is an integer from 2 to 4. 
     
     
         11 . A compound of any of  claims 1 - 10 , wherein the side chain amine group is protected by a protecting group that prevents undesired reaction of the amino group and is removable by a chemical method that does not also cause cleavage of other groups. 
     
     
         12 . A compound of any of  claims 1 - 11 , wherein the side chain carboxyl group is protected by a protecting group that prevents undesired reaction of the carboxyl group and is removable by a chemical method that does not also cause cleavage of other groups. 
     
     
         13 . A compound of  claim 11  or  12 , wherein the protecting group is BOC (t-butoxy carbonyl), FMOC (fluorenylmethoxycarbonyl), Alloc (allyloxycarbonyl), CBZ (benzyloxycarbonyl), Pbf (2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl), NO 2  (nitro), Pmc (2,2,5,7,8-pentamethylchroman-6-sulfonyl), Mtr (4-methoxy-2,3,6-trimethylbenzenesulfonyl), or Tos (tosyl). 
     
     
         14 . A semisynthetic peptide comprising a non-natural amino acid compound of any of  claims 1 - 13 . 
     
     
         15 . The semisynthetic peptide of  claim 14  comprising a non-natural amino acid compound of any of  claims 1 - 13  as its N-terminus moiety. 
     
     
         16 . The semisynthetic peptide of  claim 15  which comprises a non-natural amino acid compound of any of  claims 1 - 13  as the N-terminal moiety of a semisynthetic peptide of neurotensin (8-13). 
     
     
         17 . The semisynthetic peptide of  claim 16  which is ABS41, ABS44, ABS46, ABS201, ABS202, or ABS203. 
     
     
         18 . The semisynthetic peptide of  claim 15  which comprises a non-natural amino acid compound of any of  claims 1 - 13  as the N-terminal moiety of a semisynthetic peptide of a transcription factor, a ligand for an cellular receptor, a hormone or an extracellular binding peptide. 
     
     
         19 . The semisynthetic peptide of  claim 15  which comprises a non-natural amino acid compound of any of  claims 1 - 13  as the N-terminal moiety of a semisynthetic peptide of ofenkephlin, LHRH or analogs thereof, a neuropeptide, a glycoincretin, integrin or analogs thereof, a glucagon, a glucagon-like peptide, an antithrombotic peptide, a cytokine, an interleukin, a transferrin, an interferon, an endothelin, a natriuretic hormone, an extracellular kinase ligand, an angiotensin enzyme inhibitor, a peptide antiviral compound, thrombin, substance P, substance G, somatotropin, somatostatin, GnRH or analogues thereof, secretin, bradykinin, vasopressin or analogues thereof, insulin or analogs thereof, neurotensin, proinsulin, or a growth factor. 
     
     
         20 . The semisynthetic peptide of any of  claims 14 - 19  which has an extended half-life in vivo and/or in vitro as compared to a peptide having the same sequence as the semisynthetic peptide that does not comprise the non-natural amino acid compound substituted as its N-terminus moiety. 
     
     
         21 . A pharmaceutical composition comprising a peptide according to any of  claims 14 - 20  and a pharmaceutical carrier. 
     
     
         22 . A pharmaceutical composition according to  claim 21  wherein the peptide is present in unit dosage form. 
     
     
         23 . A cosmetic formulation comprising a desamino alkyl amino acid compound of any of  claims 1 - 13  and a cosmetic base formulation. 
     
     
         24 . A cosmetic formulation according to  claim 23  wherein the cosmetic base formulation is an aqueous or oil base. 
     
     
         25 . A cosmetic formulation comprising a semisynthetic peptide of any of  claims 14 - 20  and a cosmetic base formulation. 
     
     
         26 . A cosmetic formulation according to  claim 25  wherein the cosmetic base formulation is an aqueous or oil base. 
     
     
         27 . A compound of any one of  claims 1 - 13  for use in medical therapy. 
     
     
         28 . The use of a compound of any of  claims 1 - 13  for the manufacture of a medicament useful for treating psychosis in a mammal. 
     
     
         29 . The use of  claim 28 , wherein the psychosis is schizophrenia. 
     
     
         30 . The use of a compound of any of  claims 1 - 13  for the manufacture of a medicament useful for treating cancer in a mammal. 
     
     
         31 . The use of a compound of any of  claims 1 - 13  for the manufacture of a medicament useful for treating pain in a mammal. 
     
     
         32 . A peptide of any of  claims 14 - 20  for use in medical therapy. 
     
     
         33 . The use of a peptide of any of  claims 14 - 20  for the manufacture of a medicament useful for treating psychosis in a mammal. 
     
     
         34 . The use of  claim 33 , wherein the psychosis is schizophrenia. 
     
     
         35 . The use of a peptide of any of  claims 14 - 20  for the manufacture of a medicament useful for treating cancer in a mammal. 
     
     
         36 . The use of a peptide of any of  claims 14 - 20  for the manufacture of a medicament useful for treating pain in a mammal. 
     
     
         37 . A method to lower the body temperature of a patient, comprising administering to the patient an effective amount of a peptide of any of  claims 14 - 20  so as to lower the body temperature of the patient. 
     
     
         38 . A method to lower the body temperature of a patient, comprising administering to the patient an effective amount of a composition of any of  claims 21 - 22  so as to lower the body temperature of the patient. 
     
     
         39 . A method to treat cancer, comprising administering to a patient an effective amount of a peptide of any of  claims 14 - 20  so as to treat the cancer. 
     
     
         40 . A method to treat cancer, comprising administering to a patient an effective amount of a composition of any of  claims 21 - 22  so as to treat the cancer. 
     
     
         41 . A method to treat pain, comprising administering to a patient an effective amount of a peptide of any of  claims 14 - 20  so as to treat the pain. 
     
     
         42 . A method to treat pain, comprising administering to a patient an effective amount of a composition of any of  claims 21 - 22  so as to treat the pain. 
     
     
         43 . A method to treat a patient with psychosis, comprising administering to the patient an effective amount of a peptide of any of  claims 14 -20 so as to treat the psychosis. 
     
     
         44 . A method to treat a patient with psychosis, comprising administering to the patient an effective amount of a composition of any of  claims 21 - 22  so as to treat the psychosis. 
     
     
         45 . A method for screening a peptide containing a non-natural amino acid compound for an activity, comprising the steps of:
 a) measuring a biological activity of a first peptide having a known amino acid sequence; and   b) measuring the same biological activity of a semisynthetic peptide of any of  claims 14 - 20  wherein the semisynthetic peptide has the same sequence as the first peptide except for the non-natural amino acid compound, or is a truncated version of the first peptide except for the non-natural amino acid compound.   
     
     
         46 . The method of  claim 45 , wherein the biological activity is poptosis, apoptosis, cell signaling, ligand binding, transcription, translation, metabolism, cell growth, cell differentiation, homeostasis, half-life, solubility, transport, or stability. 
     
     
         47 . The method of  claim 45 , wherein the biological activity includes a direct or indirect assessment of the ability of the semisynthetic peptide to pass through a biological barrier. 
     
     
         48 . The method of  claim 45 , wherein the biological activity is selectivity. 
     
     
         49 . A method of treating a patient with a disease that is affected by administration to the patient of a known first peptide, comprising administering to the patient a semisynthetic peptide of any of  claims 14 - 20  wherein the semisynthetic peptide has the same sequence as the first peptide except for the non-natural amino acid compound, or is a truncated version of the first peptide except for the non-natural amino acid compound. 
     
     
         50 . A method of increasing the ability of a known first peptide to cross a biological barrier of a subject, comprising substituting a semisynthetic peptide of any of  claims 14 - 20  wherein the semisynthetic peptide has the same sequence as the first peptide except for the non-natural amino acid compound, or is a truncated version of the first peptide except for the non-natural amino acid compound. 
     
     
         51 . The method of  claim 50 , wherein the barrier comprises the blood brain barrier, a cell membrane, intestinal epithelium, skin, or blood-ocular. 
     
     
         52 . The method of  claim 50 , wherein the barrier is the blood brain barrier. 
     
     
         53 . A method of increasing the selectivity of a known peptide, comprising substituting for the known peptide a semisynthetic peptide of any of  claims 14 - 20  wherein the semisynthetic peptide has the same sequence as the first peptide except for the non-natural amino acid compound, or is a truncated version of the first peptide except for the non-natural amino acid compound. 
     
     
         54 . A method of increasing the resistance of a known peptide to digestion by a peptidase, comprising substituting for the known peptide a semisynthetic peptide of any of  claims 14 - 20  wherein the semisynthetic peptide has the same sequence as the first peptide except for the non-natural amino acid compound, or is a truncated version of the first peptide except for the non-natural amino acid compound. 
     
     
         55 . A method of treating a patient with a disease that is affected by administration to the patient of a known first peptide that crosses a body barrier, comprising administering to the patient a semisynthetic peptide of any of  claims 14 - 20  wherein the semisynthetic peptide has the same sequence as the first peptide except for the non-natural amino acid compound, or is a truncated version of the first peptide except for the non-natural amino acid compound. 
     
     
         56 . A method of treating a patient with a disease of the brain that is affected by administration to the patient of a known first peptide, comprising administering to the patient a semisynthetic peptide of any of  claims 14 - 20  wherein the semisynthetic peptide has the same sequence as the first peptide except for the non-natural amino acid compound, or is a truncated version of the first peptide except for the non-natural amino acid compound. 
     
     
         57 . A method for preparing a semisynthetic peptide with an extended half-life in vivo comprising substituting for a known peptide a semisynthetic peptide of any of  claims 14 - 20  wherein the semisynthetic peptide has the same sequence as the first peptide except for the non-natural amino acid compound, or is a truncated version of the first peptide except for the non-natural amino acid compound.