US2022288062A1PendingUtilityA1

Treatment of t-cell mediated diseases

84
Assignee: AMPIO PHARMACEUTICALS INCPriority: May 15, 2003Filed: May 23, 2022Published: Sep 15, 2022
Est. expiryMay 15, 2023(expired)· nominal 20-yr term from priority
A61P 25/02A61P 43/00A61P 1/00A61K 9/0048A61P 13/00A61P 11/00A61P 27/02A61P 17/00A61P 1/16A61P 5/50A61P 1/04A61P 19/02A61K 38/12A61K 9/48A61P 35/00A61K 9/00A61P 19/04A61P 37/08A61K 31/4965C07D 241/08A61P 37/06A61P 1/18A61P 13/12A61K 31/495A61P 25/00A61K 45/06A61P 37/02A61P 21/04A61P 37/00A61K 9/20A61P 29/00A61P 21/00A61P 7/06A61P 5/38A61P 17/06A61P 3/10A61P 5/14A61P 9/10
84
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Claims

Abstract

The invention provides a method of treating T-cell mediated diseases and a method of inhibiting the activation of T-cells using certain diketopiperazines. The invention also provides methods of synthesizing diketopiperazines and pharmaceutical compositions comprising certain diketopiperazines. The invention further provides methods of making improved pharmaceutical compositions of proteins and peptides by either increasing or decreasing the content of diketopiperazines in the compositions and the resultant improved pharmaceutical compositions.

Claims

exact text as granted — not AI-modified
We claim: 
     
         1 . A method of treating a T-cell mediated disease comprising administering to an animal in need thereof an effective amount of a diketopiperazine having the following formula: 
       
         
           
           
               
               
           
         
       
       wherein:
 R 1  and R 2 , which may be the same or different, each is: 
 (a) a side chain of an amino acid, wherein the amino acid is glycine, alanine, valine, norvaline, α-aminoisobutyric acid, 2,4-diaminobutyric acid, 2,3-diaminobutyric acid, leucine, isoleucine, norleucine, serine, homoserine, threonine, aspartic acid, asparagine, glutamic acid, glutamine, lysine, hydroxylysine, histidine, arginine, homoarginine, citrulline, phenylalanine, p-aminophenylalanine, tyrosine, tryptophan, thyroxine, cysteine, homocysteine, methionine, penicillamine or ornithine; provided, however, that when R 1  is the side chain of asparagine or glutamine, then R 2  cannot be the side chain of lysine or ornithine, and when R 1  is the side chain of lysine or ornithine, then R 2  cannot be the side chain of asparagine or glutamine; 
 (b) R 1  is —CH 2 —CH 2 —CH 2 — or —CH 2 —CH(OH)—CH 2 — and together with the adjacent ring nitrogen forms proline or hydroxyproline, R 2  is —CH 2 —CH 2 —CH 2 — or —CH 2 —CH(OH)—CH 2 — and together with the adjacent ring nitrogen forms proline or hydroxyproline, or both R 1  and R 2  are each independently —CH 2 —CH 2 —CH 2 — or —CH 2 —CH(OH)—CH 2 — and together with the adjacent ring nitrogens form proline or hydroxyproline; or 
 (c) a derivative of a side chain of an amino acid, wherein the amino acid is one of those recited in (a), and the derivatized side chain has:
 (i) an —NH 2  group replaced by an —NHR 3  or —N(R 3 ) 2  group, wherein each R 3  may independently be a substituted or unsubstituted alkyl, cycloalkyl, heterocycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl; 
 (ii) an —OH group replaced by an —O—PO 3 H 2  or —OR 3  group, wherein each R 3  may independently be a substituted or unsubstituted alkyl, cycloalkyl, heterocycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl; 
 (iii) a —COOH group replaced by a —COOR 3  group, wherein each R 3  may independently be a substituted or unsubstituted alkyl, cycloalkyl, heterocycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl; 
 (iv) a —COOH group replaced by a —CON(R 4 ) 2  group, wherein each R 4  may independently be H or a substituted or unsubstituted alkyl, cycloalkyl, heterocycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl; 
 (v) an —SH group replaced by —S—S—CH 2 —CH(NH 2 )—COOH or —S—S—CH 2 —CH 2 —CH(NH 2 )—COOH; 
 (vi) a —CH 2 — group replaced by a —CH(NH 2 )— or a —CH(OH)— group; 
 (vii) a —CH 3  group replaced by a —CH 2 —NH 2  or a —CH 2 —OH group; and/or 
 (viii) an H which is attached to a carbon atom replaced by a halogen; or 
 
 
       a physiologically-acceptable salt thereof. 
     
     
         2 . The method of  claim 1  wherein R 1 , R 2  or both is the side chain of aspartic acid, the side chain of glutamic acid, or a derivative of a side chain of aspartic acid or glutamic acid wherein the —COOH group is replaced by a —COOR 3  group or a —CON(R 4 ) 2  group. 
     
     
         3 . The method of  claim 2  wherein R 1  is the side chain of aspartic acid or a derivative of the side chain of aspartic acid wherein the —COOH group is replaced by a —COOR 3  group or a —CON(R 4 ) 2  group, and R 2  is the side chain of alanine. 
     
     
         4 . The method of  claim 2  wherein R 1  is the side chain of aspartic acid or a derivative of the side chain of aspartic acid wherein the —COOH group is replaced by a —COOR 3  group or a —CON(R 4 ) 2  group, and R 2  is the side chain of tyrosine. 
     
     
         5 . The method of  claim 2  wherein R 1  is the side chain of glutamic acid or a derivative of the side chain of glutamic acid wherein the —COOH group is replaced by a —COOR 3  group or a —CON(R 4 ) 2  group, and R 2  is the side chain of alanine. 
     
     
         6 . The method of  claim 2  wherein R 1  is the side chain of glutamic acid or a derivative of the side chain of glutamic acid wherein the —COOH group is replaced by a —COOR 3  group or a —CON(R 4 ) 2  group, and R 2  is the side chain of tyrosine. 
     
     
         7 . The method of  claim 2  wherein R 1  is the side chain of aspartic acid or glutamic acid and R 2  is the side chain of alanine. 
     
     
         8 . The method of  claim 2  wherein R 1  is the side chain of aspartic acid or glutamic acid and R 2  is the side chain of tyrosine. 
     
     
         9 . The method of  claim 1  wherein R 1  and R 2  are both a hydrophobic side chain or a hydrophobic side chain derivative. 
     
     
         10 . The method of  claim 9  wherein:
 (a) R 1  and R 2 , which may be the same or different, each is the side chain of glycine, alanine, valine, norvaline, α-aminobutyric acid, leucine, isoleucine, norleucine or phenylalanine; 
 (b) R 1  is —CH 2 —CH 2 —CH 2 — and together with the adjacent nitrogen atom forms proline, and R 2  is —CH 2 —CH 2 —CH 2 — and together with the adjacent nitrogen atom forms proline; or 
 (c) R 1  is the side chain of glycine, alanine, valine, norvaline, α-aminobutyric acid, leucine, isoleucine, norleucine or phenylalanine, and R 2  is —CH 2 —CH 2 —CH 2 — and together with the adjacent nitrogen atom forms proline. 
 
     
     
         11 . The method of  claim 10  wherein R 1  is the side chain of glycine and R 2  is the side chain of leucine. 
     
     
         12 . The method of  claim 10  wherein R 1  is —CH 2 —CH 2 —CH 2 — and together with the adjacent nitrogen atom forms proline and R 2  is the side chain of phenylalanine. 
     
     
         13 . The method of  claim 10  wherein R 1  is —CH 2 —CH 2 —CH 2 — and together with the adjacent nitrogen atom forms proline and R 2  is the side chain of alanine. 
     
     
         14 . The method of  claim 1  wherein R 1 , R 2  or both is the side chain of methionine, the side chain of arginine or a derivative of these side chains. 
     
     
         15 . The method of  claim 14  wherein R 1  is the side chain of methionine and R 2  is the side chain of arginine. 
     
     
         16 . The method of any one of  claims 1 - 15  wherein the animal is a human. 
     
     
         17 . The method of any one of  claims 1 - 15  wherein the T-cell mediated disease is graft rejection, graft versus host disease, an unwanted delayed-type hypersensitivity reaction, a T-cell mediated pulmonary disease or an autoimmune disease. 
     
     
         18 . The method of any one of  claims 1 - 15  wherein the T-cell mediated disease is multiple sclerosis, neuritis, polymyositis, psoriasis, vitiligo, Sjogren's syndrome, rheumatoid arthritis, Type 1 diabetes, autoimmune pancreatitis, inflammatory bowel diseases, Crohn's disease, ulcerative colitis, celiac disease, glomerulonephritis, scleroderma, sarcoidosis, autoimmune thyroid diseases, Hashimoto's thyroiditis, Graves disease, myasthenia gravis, Addison's disease, autoimmune uveoretinitis, pemphigus vulgaris, primary biliary cirrhosis, pernicious anemia, or systemic lupus erythematosis. 
     
     
         19 . The method of any one of  claims 1 - 15  wherein the T-cell mediated disease is pulmonary fibrosis or idiopathic pulmonary fibrosis. 
     
     
         20 . A method of inhibiting activation of T-cells comprising administering to an animal in need thereof an effective amount of a diketopiperazine having the following formula: 
       
         
           
           
               
               
           
         
       
       wherein:
 R 1  and R 2 , which may be the same or different, each is: 
 (a) a side chain of an amino acid, wherein the amino acid is glycine, alanine, valine, norvaline, α-aminoisobutyric acid, 2,4-diaminobutyric acid, 2,3-diaminobutyric acid, leucine, isoleucine, norleucine, serine, homoserine, threonine, aspartic acid, asparagine, glutamic acid, glutamine, lysine, hydroxylysine, histidine, arginine, homoarginine, citrulline, phenylalanine, p-aminophenylalanine, tyrosine, tryptophan, thyroxine, cysteine, homocysteine, methionine, penicillamine or ornithine; provided, however, that when R 1  is the side chain of asparagine or glutamine, then R 2  cannot be the side chain of lysine or ornithine, and when R 1  is the side chain of lysine or ornithine, then R 2  cannot be the side chain of asparagine or glutamine; 
 (b) R 1  is —CH 2 —CH 2 —CH 2 — or —CH 2 —CH(OH)—CH 2 — and together with the adjacent ring nitrogen forms proline or hydroxyproline, R 2  is —CH 2 —CH 2 —CH 2 — or —CH 2 —CH(OH)—CH 2 — and together with the adjacent ring nitrogen forms proline or hydroxyproline, or both R 1  and R 2  are each independently —CH 2 —CH 2 —CH 2 — or —CH 2 —CH(OH)—CH 2 — and together with the adjacent ring nitrogens form proline or hydroxyproline; or 
 (c) a derivative of a side chain of an amino acid, wherein the amino acid is one of those recited in (a), and the derivatized side chain has:
 (i) an —NH 2  group replaced by an —NHR 3  or —N(R 3 ) 2  group, wherein each R 3  may independently be a substituted or unsubstituted alkyl, cycloalkyl, heterocycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl; 
 (ii) an —OH group replaced by an —O—PO 3 H 2  or —OR 3  group, wherein each R 3  may independently be a substituted or unsubstituted alkyl, cycloalkyl, heterocycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl; 
 (iii) a —COOH group replaced by a —COOR 3  group, wherein each R 3  may independently be a substituted or unsubstituted alkyl, cycloalkyl, heterocycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl; 
 (iv) a —COOH group replaced by a —CON(R 4 ) 2  group, wherein each R 4  may independently be H or a substituted or unsubstituted alkyl, cycloalkyl, heterocycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl; 
 (v) an —SH group replaced by —S—S—CH 2 —CH(NH 2 )—COOH or —S—S—CH 2 —CH 2 —CH(NH 2 )—COOH; 
 (vi) a —CH 2 — group replaced by a —CH(NH 2 )— or a —CH(OH)— group; 
 (vii) a —CH 3  group replaced by a —CH 2 —NH 2  or a —CH 2 —OH group; and/or 
 (viii) an H which is attached to a carbon atom replaced by a halogen; or 
 
 
       a physiologically-acceptable salt thereof. 
     
     
         21 . The method of  claim 20  wherein R 1 , R 2  or both is the side chain of aspartic acid, the side chain of glutamic acid, or a derivative of a side chain of aspartic acid or glutamic acid wherein the —COOH group is replaced by a —COOR 3  group or a —CON(R 4 ) 2  group. 
     
     
         22 . The method of  claim 21  wherein R 1  is the side chain of aspartic acid or a derivative of the side chain of aspartic acid wherein the —COOH group is replaced by a —COOR 3  group or a —CON(R 4 ) 2  group, and R 2  is the side chain of alanine. 
     
     
         23 . The method of  claim 21  wherein R 1  is the side chain of aspartic acid or a derivative of the side chain of aspartic acid wherein the —COOH group is replaced by a —COOR 3  group or a —CON(R 4 ) 2  group, and R 2  is the side chain of tyrosine. 
     
     
         24 . The method of  claim 21  wherein R 1  is the side chain of glutamic acid or a derivative of the side chain of glutamic acid wherein the —COOH group is replaced by a —COOR 3  group or a —CON(R 4 ) 2  group, and R 2  is the side chain of alanine. 
     
     
         25 . The method of  claim 21  wherein R 1  is the side chain of glutamic acid or a derivative of the side chain of glutamic acid wherein the —COOH group is replaced by a —COOR 3  group or a —CON(R 4 ) 2  group, and R 2  is the side chain of tyrosine. 
     
     
         26 . The method of  claim 21  wherein R 1  is the side chain of aspartic acid or glutamic acid and R 2  is the side chain of alanine. 
     
     
         27 . The method of  claim 21  wherein R 1  is the side chain of aspartic acid or glutamic acid and R 2  is the side chain of tyrosine. 
     
     
         28 . The method of  claim 20  wherein R 1  and R 2  are both a hydrophobic side chain or a hydrophobic side chain derivative. 
     
     
         29 . The method of  claim 28  wherein:
 (a) R 1  and R 2 , which may be the same or different, each is the side chain of glycine, alanine, valine, norvaline, α-aminobutyric acid, leucine, isoleucine, norleucine or phenylalanine; 
 (b) R 1  is —CH 2 —CH 2 —CH 2 — and together with the adjacent nitrogen atom forms proline, and R 2  is —CH 2 —CH 2 —CH 2 — and together with the adjacent nitrogen atom forms proline; or 
 (c) R 1  is the side chain of glycine, alanine, valine, norvaline, α-aminobutyric acid, leucine, isoleucine, norleucine or phenylalanine, and R 2  is —CH 2 —CH 2 —CH 2 — and together with the adjacent nitrogen atom forms proline. 
 
     
     
         30 . The method of  claim 29  wherein R 1  is the side chain of glycine and R 2  is the side chain of leucine. 
     
     
         31 . The method of  claim 29  wherein R′ is —CH 2 —CH 2 —CH 2 — and together with the adjacent nitrogen atom forms proline and R 2  is the side chain of phenylalanine. 
     
     
         32 . The method of  claim 29  wherein R′ is —CH 2 —CH 2 —CH 2 — and together with the adjacent nitrogen atom forms proline and R 2  is the side chain of alanine. 
     
     
         33 . The method of  claim 20  wherein R′, R 2  or both is the side chain of methionine, the side chain of arginine or a derivative of these side chains. 
     
     
         34 . The method of  claim 33  wherein R 1  is the side chain of methionine and R 2  is the side chain of arginine. 
     
     
         35 . The method of any one of  claims 20 - 34  wherein the animal is a human. 
     
     
         36 . The method of any one of  claims 20 - 34  wherein the diketopiperazine is used to treat inflammation or an inflammatory disease which is caused or exacerbated at least in part by T-cell activation. 
     
     
         37 . A pharmaceutical composition comprising a pharmaceutically-acceptable carrier and a diketopiperazine having the following formula: 
       
         
           
           
               
               
           
         
       
       wherein:
 R 5  and R 6 , which may be the same or different, each is: 
 (a) a side chain of an amino acid, wherein the amino acid is glycine, alanine, valine, norvaline, α-aminoisobutyric acid, 2,4-diaminobutyric acid, 2,3-diaminobutyric acid, leucine, isoleucine, norleucine, serine, homoserine, threonine, lysine, hydroxylysine, histidine, arginine, homoarginine, citrulline, phenylalanine, p-aminophenylalanine, tyrosine, tryptophan, thyroxine or ornithine; provided, however, that when R 5  is the side chain of asparagine or glutamine, then R 6  cannot be the side chain of lysine or ornithine, and when R 5  is the side chain of lysine or ornithine, then R 6  cannot be the side chain of asparagine or glutamine; 
 (b) R 5  is —CH 2 —CH 2 —CH 2 — or —CH 2 —CH(OH)—CH 2 — and together with the adjacent ring nitrogen forms proline or hydroxyproline, R 6  is —CH 2 —CH 2 —CH 2 — or —CH 2 —CH(OH)—CH 2 — and together with the adjacent ring nitrogen forms proline or hydroxyproline, or both R 5  and R 6  are each independently —CH 2 —CH 2 —CH 2 — or —CH 2 —CH(OH)—CH 2 — and together with the adjacent ring nitrogens form proline or hydroxyproline; or 
 (c) a derivative of a side chain of an amino acid, wherein the amino acid is one of those recited in (a), and the derivatized side chain has:
 (i) an —NH 2  group replaced by an —NHR 3  or —N(R 3 ) 2  group, wherein each R 3  may independently be a substituted or unsubstituted alkyl, cycloalkyl, heterocycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl; 
 (ii) an —OH group replaced by an —O—PO 3 H 2  or —OR 3  group, wherein each R 3  may independently be a substituted or unsubstituted alkyl, cycloalkyl, heterocycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl; 
 (iii) a —CH 2 — group replaced by a —CH(NH 2 )— or a —CH(OH)— group; 
 (iv) a —CH 3  group replaced by a —CH 2 —NH 2  or a —CH 2 —OH group; and/or 
 (v) an H which is attached to a carbon atom replaced by a halogen; or 
 
 
       a physiologically-acceptable salt thereof. 
     
     
         38 . The composition of  claim 37  wherein R 5  and R 6  are both a hydrophobic side chain or a hydrophobic side chain derivative. 
     
     
         39 . The composition of  claim 38  wherein:
 (a) R 5  and R 6 , which may be the same or different, each is the side chain of glycine, alanine, valine, norvaline, α-aminobutyric acid, leucine, isoleucine, norleucine or phenylalanine; 
 (b) R 5  is —CH 2 —CH 2 —CH 2 — and together with the adjacent nitrogen atom forms proline, and R 6  is —CH 2 —CH 2 —CH 2 — and together with the adjacent nitrogen atom forms proline; or 
 (c) R 5  is the side chain of glycine, alanine, valine, norvaline, α-aminobutyric acid, leucine, isoleucine, norleucine or phenylalanine, and R 6  is —CH 2 —CH 2 —CH 2 — and together with the adjacent nitrogen atom forms proline. 
 
     
     
         40 . The composition of  claim 39  wherein R 5  is the side chain of glycine and R 6  is the side chain of leucine. 
     
     
         41 . The composition of  claim 39  wherein R 5  is —CH 2 —CH 2 —CH 2 — and together with the adjacent nitrogen atom forms proline and R 6  is the side chain of phenylalanine. 
     
     
         42 . The composition of  claim 39  wherein R 5  is —CH 2 —CH 2 —CH 2 — and together with the adjacent nitrogen atom forms proline and R 6  is the side chain of alanine. 
     
     
         43 . The composition of  claim 37  wherein R 5 , R 6  or both is the side chain of methionine, the side chain of arginine or a derivative of these side chains. 
     
     
         44 . The composition of  claim 43  wherein R 5  is the side chain of methionine and R 6  is the side chain of arginine. 
     
     
         45 . A method of treating a T-cell mediated disease comprising administering to an animal in need thereof an effective amount of a pharmaceutical composition comprising a protein or peptide normally found in the animal, the protein or peptide having been treated so that the composition also comprises at least one diketopiperazine derived from the protein or peptide. 
     
     
         46 . The method of  claim 45  where in the protein is albumin. 
     
     
         47 . The method of  claim 45  wherein the protein is immunoglobulin. 
     
     
         48 . The method of  claim 45  wherein the protein is erythropoietin. 
     
     
         49 . The method of any one of  claims 45 - 48  wherein the pharmaceutical composition is administered orally. 
     
     
         50 . The method of any one of  claims 45 - 48  wherein the animal is a human and the protein or peptide is a human protein or peptide. 
     
     
         51 . A method of inhibiting T-cell activation comprising administering to an animal in need thereof an effective amount of a pharmaceutical composition comprising a protein or peptide normally found in the animal, the protein or peptide having been treated so that the composition also comprises at least one diketopiperazine derived from the protein or peptide. 
     
     
         52 . The method of  claim 51  where in the protein is albumin. 
     
     
         53 . The method of  claim 51  wherein the protein is immunoglobulin. 
     
     
         54 . The method of  claim 51  wherein the protein is erythropoietin. 
     
     
         55 . The method of any one of  claims 51 - 54  wherein the pharmaceutical composition is administered orally. 
     
     
         56 . The method of any one of  claims 51 - 54  wherein the animal is a human and the protein or peptide is a human protein or peptide. 
     
     
         57 . A method of synthesizing a diketopiperazine comprising heating a solution of a protein or peptide under conditions effective to cause the formation of the diketopiperazine. 
     
     
         58 . The method of  claim 57  wherein the protein is albumin. 
     
     
         59 . The method of  claim 57  wherein the protein is an immunoglobulin. 
     
     
         60 . The method of  claim 57  wherein the protein is erythropoietin. 
     
     
         61 . The method of  claim 57  wherein the diketopiperazine is purified from the solution. 
     
     
         62 . The method of any one of  claims 57 - 61  wherein the solution is heated for four days at 60° C. 
     
     
         63 . A method of synthesizing a diketopiperazine comprising contacting a solution of a protein or peptide with an enzyme that cleaves the two N-terminal or the two C-terminal amino acids of the protein or peptide under conditions effective to produce the diketopiperazine. 
     
     
         64 . The method of  claim 63  wherein the protein is albumin. 
     
     
         65 . The method of  claim 63  wherein the protein is an immunoglobulin. 
     
     
         66 . The method of  claim 63  wherein the protein is erythropoietin. 
     
     
         67 . The method of  claim 63  wherein the enzyme is a dipeptidyl peptidase 
     
     
         68 . The method of  claim 63  wherein the enzyme is a carboxypeptidase. 
     
     
         69 . The method of any one of  claims 63 - 68  wherein the diketopiperazine is purified from the solution. 
     
     
         70 . An improved pharmaceutical composition of a protein or peptide, the improvement comprising a decreased content of diketopiperazines in the composition. 
     
     
         71 . The composition of  claim 70  wherein the protein is albumin. 
     
     
         72 . The composition of  claim 70  wherein the protein is an immunoglobulin. 
     
     
         73 . The composition of  claim 70  wherein the protein is erythropoietin. 
     
     
         74 . A method of making an improved pharmaceutical composition of a protein or peptide, the method comprising removing from the composition at least some of the diketopiperazines present in the composition. 
     
     
         75 . The composition of  claim 74  wherein the protein is albumin. 
     
     
         76 . The composition of  claim 74  wherein the protein is an immunoglobulin. 
     
     
         77 . The composition of  claim 74  wherein the protein is erythropoietin. 
     
     
         78 . A method of making an improved pharmaceutical composition of a protein or peptide, the method comprising treating a solution of the protein or peptide so as to increase the content of diketopiperazines. 
     
     
         79 . The method of  claim 78  wherein the solution is heated under conditions effective to cause the formation of diketopiperazines 
     
     
         80 . The method of  claim 79  wherein the solution is heated for four days at 60° C. 
     
     
         81 . The method of  claim 78  wherein the solution is contacted with an enzyme that cleaves the two N-terminal or the two C-terminal amino acids of the protein or peptide under conditions effective to produce the diketopiperazines. 
     
     
         82 . The method of  claim 81  wherein the enzyme is a dipeptidyl peptidase 
     
     
         83 . The method of  claim 81  wherein the enzyme is a carboxypeptidase. 
     
     
         84 . The method of  claim 78  wherein the protein is albumin. 
     
     
         85 . The method of  claim 78  wherein the protein is an immunoglobulin. 
     
     
         86 . The method of  claim 78  wherein the protein is erythropoietin. 
     
     
         87 . An improved pharmaceutical composition of a protein or peptide, the improvement comprising an increased content of diketopiperazines in the composition. 
     
     
         88 . The composition of  claim 87  wherein the protein is albumin. 
     
     
         89 . The composition of  claim 87  wherein the protein is an immunoglobulin. 
     
     
         90 . The composition of  claim 87  wherein the protein is erythropoietin. 
     
     
         91 . The composition of any one of  claims 87 - 90  which is suitable for oral administration.

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