Methods and compositions targeting viral and cellular ITAM motifs, and use of same in identifying compounds with therapeutic activity

Abstract

This invention provides methods of treating, reducing the incidence of, and inhibiting metastasis formation of carcinomas, sarcomas, Epstein-Barr virus-induced malignancies, B cell proliferative disorders, and mast cell activation disorders, comprising administering to a subject a compound that inhibits an interaction of a first protein and an immunoreceptor tyrosine-based activation motif (ITAM) of a second protein, and screening methods for identifying ITAM-inhibitory compounds and peptides. This invention also provides peptides that inhibit signaling by ITAMs.

1 . A method of treating a breast cell carcinoma in a subject, comprising administering to said subject a compound that inhibits an interaction of a first protein and an immunoreceptor tyrosine-based activation motif (ITAM) of a second protein, thereby treating a breast cell carcinoma in a subject.  
     
     
         2 . The method of  claim 1 , wherein said compound comprises a peptide homologous to said ITAM.  
     
     
         3 . The method of  claim 2 , wherein said peptide comprises a modification selected from hydroxylation, amidation, esterification, formylation, gamma-carboxyglutamic acid hydroxylation, methylation, phosphorylation, sulfation, glycosylation, reduction, oxidation, disulfide modification, introduction of a thioether bond, introduction of a thiolester bond, a backbone condensation, or a biotinylation.  
     
     
         4 . The method of  claim 2 , wherein said peptide comprises an amino acid selected from a D amino acid; pyrrolidone carboxylic acid; 2-aminoadipic acid; 3-aminoadipic acid; beta-alanine; beta-aminoproprionic acid; 2-aminobutyric acid; 4-aminobutyric acid; piperidinic acid; 6-aminocaproic acid; 6-aminoheptanoic acid; 2-aminoheptanoic acid; 2-aminoisobutyric acid; 3-aminoisobutyric acid; 2-aminopimelic acid; 2,4 diaminobutyric acid; desmosine; 2,2 diaminopimelic acid; 2,3 diaminopropionic acid; N-ethylglycine; N-ethylasparagine; hydroxylysine; allo-hydroxylysine; 3-hydroxyproline; 4-hydroxyproline; isodesmosine; allo-isoleucine; N-methylglycine; sarcosine; methylisoleucine; methyllysine; methylvaline; norvaline; norleucine; 6-aminohexanoic acid; citrulline; cysteic acid; cyclohexylalanine; alpha-amino isobutyric acid; t-butylglycine; t-butylalanine; phenylglycine; an N-alpha-methyl amino acid, a C-alpha-methyl amino acid, a beta-methyl amino acid, or orthinine.  
     
     
         5 . The method of  claim 1 , wherein said second protein is a viral protein.  
     
     
         6 . A method of reducing an incidence of a breast cell carcinoma in a subject, comprising administering to said subject a compound that inhibits an interaction of a first protein and an immunoreceptor tyrosine-based activation motif (ITAM) of a second protein, thereby reducing an incidence of a breast cell carcinoma in a subject.  
     
     
         7 . The method of  claim 6 , wherein said compound comprises a peptide homologous to said ITAM.  
     
     
         8 . The method of  claim 7 , wherein said peptide comprises a modification selected from hydroxylation, amidation, esterification, formylation, gamma-carboxyglutamic acid hydroxylation, methylation, phosphorylation, sulfation, glycosylation, reduction, oxidation, disulfide modification, introduction of a thioether bond, introduction of a thiolester bond, a backbone condensation, or a biotinylation.  
     
     
         9 . The method of  claim 7 , wherein said peptide comprises an amino acid selected from a D amino acid; pyrrolidone carboxylic acid; 2-aminoadipic acid; 3-aminoadipic acid; beta-alanine; beta-aminoproprionic acid; 2-aminobutyric acid; 4-aminobutyric acid; piperidinic acid; 6-aminocaproic acid; 6-aminoheptanoic acid; 2-aminoheptanoic acid; 2-aminoisobutyric acid; 3-aminoisobutyric acid; 2-aminopimelic acid; 2,4 diaminobutyric acid; desmosine; 2,2 diaminopimelic acid; 2,3 diaminopropionic acid; N-ethylglycine; N-ethylasparagine; hydroxylysine; allo-hydroxylysine; 3-hydroxyproline; 4-hydroxyproline; isodesmosine; allo-isoleucine; N-methylglycine; sarcosine; methylisoleucine; methyllysine; methylvaline; norvaline; norleucine; 6-aminohexanoic acid; citrulline; cysteic acid; cyclohexylalanine; alpha-amino isobutyric acid; t-butylglycine; t-butylalanine; phenylglycine; an N-alpha-methyl amino acid, a C-alpha-methyl amino acid, a beta-methyl amino acid, or orthinine.  
     
     
         10 . The method of  claim 6 , wherein said second protein is a viral protein.  
     
     
         11 . A method of inhibiting a formation of a metastasis of a breast cell carcinoma in a subject, comprising administering to said subject a compound that inhibits an interaction of a first protein and an immunoreceptor tyrosine-based activation motif (ITAM) of a second protein, thereby inhibiting a formation of a metastasis of a breast cell carcinoma in a subject.  
     
     
         12 . The method of  claim 11 , wherein said compound comprises a peptide homologous to said ITAM.  
     
     
         13 . The method of  claim 12 , wherein said peptide comprises a modification selected from hydroxylation, amidation, esterification, formylation, gamma-carboxyglutamic acid hydroxylation, methylation, phosphorylation, sulfation, glycosylation, reduction, oxidation, disulfide modification, introduction of a thioether bond, introduction of a thiolester bond, a backbone condensation, or a biotinylation.  
     
     
         14 . The method of  claim 12 , wherein said peptide comprises an amino acid selected from a D amino acid; pyrrolidone carboxylic acid; 2-aminoadipic acid; 3-aminoadipic acid; beta-alanine; beta-aminoproprionic acid; 2-aminobutyric acid; 4-aminobutyric acid; piperidinic acid; 6-aminocaproic acid; 6-aminoheptanoic acid; 2-aminoheptanoic acid; 2-aminoisobutyric acid; 3-aminoisobutyric acid; 2-aminopimelic acid; 2,4 diaminobutyric acid; desmosine; 2,2 diaminopimelic acid; 2,3 diaminopropionic acid; N-ethylglycine; N-ethylasparagine; hydroxylysine; allo-hydroxylysine; 3-hydroxyproline; 4-hydroxyproline; isodesmosine; allo-isoleucine; N-methylglycine; sarcosine; methylisoleucine; methyllysine; methylvaline; norvaline; norleucine; 6-aminohexanoic acid; citrulline; cysteic acid; cyclohexylalanine; alpha-amino isobutyric acid; t-butylglycine; t-butylalanine; phenylglycine; an N-alpha-methyl amino acid, a C-alpha-methyl amino acid, a beta-methyl amino acid, or orthinine.  
     
     
         15 . The method of  claim 11 , wherein said second protein is a viral protein.  
     
     
         16 . A method of treating a fibrosarcoma or Kaposi's sarcoma in a subject, comprising administering to said subject a compound that inhibits an interaction of a first protein and an immunoreceptor tyrosine-based activation motif (ITAM) of a second protein, thereby treating a fibrosarcoma or Kaposi's sarcoma in a subject.  
     
     
         17 . The method of  claim 16 , wherein said compound comprises a peptide homologous to said ITAM.  
     
     
         18 . The method of  claim 17 , wherein said peptide comprises a modification selected from hydroxylation, amidation, esterification, formylation, gamma-carboxyglutamic acid hydroxylation, methylation, phosphorylation, sulfation, glycosylation, reduction, oxidation, disulfide modification, introduction of a thioether bond, introduction of a thiolester bond, a backbone condensation, or a biotinylation.  
     
     
         19 . The method of  claim 17 , wherein said peptide comprises an amino acid selected from a D amino acid; pyrrolidone carboxylic acid; 2-aminoadipic acid; 3-aminoadipic acid; beta-alanine; beta-aminoproprionic acid; 2-aminobutyric acid; 4-aminobutyric acid; piperidinic acid; 6-aminocaproic acid; 6-aminoheptanoic acid; 2-aminoheptanoic acid; 2-aminoisobutyric acid; 3-aminoisobutyric acid; 2-aminopimelic acid; 2,4 diaminobutyric acid; desmosine; 2,2 diaminopimelic acid; 2,3 diaminopropionic acid; N-ethylglycine; N-ethylasparagine; hydroxylysine; allo-hydroxylysine; 3-hydroxyproline; 4-hydroxyproline; isodesmosine; allo-isoleucine; N-methylglycine; sarcosine; methylisoleucine; methyllysine; methylvaline; norvaline; norleucine; 6-aminohexanoic acid; citrulline; cysteic acid; cyclohexylalanine; alpha-amino isobutyric acid; t-butylglycine; t-butylalanine; phenylglycine; an N-alpha-methyl amino acid, a C-alpha-methyl amino acid, a beta-methyl amino acid, or orthinine.  
     
     
         20 . The method of  claim 16 , wherein said second protein is a viral protein.  
     
     
         21 . A method of reducing an incidence of a fibrosarcoma or Kaposi's sarcoma in a subject, comprising administering to said subject a compound that inhibits an interaction of a first protein and an immunoreceptor tyrosine-based activation motif (ITAM) of a second protein, thereby reducing an incidence of a fibrosarcoma or Kaposi's sarcoma in a subject.  
     
     
         22 . The method of  claim 21 , wherein said compound comprises a peptide homologous to said ITAM.  
     
     
         23 . The method of  claim 22 , wherein said peptide comprises a modification selected from hydroxylation, amidation, esterification, formylation, gamma-carboxyglutamic acid hydroxylation, methylation, phosphorylation, sulfation, glycosylation, reduction, oxidation, disulfide modification, introduction of a thioether bond, introduction of a thiolester bond, a backbone condensation, or a biotinylation.  
     
     
         24 . The method of  claim 22 , wherein said peptide comprises an amino acid selected from a D amino acid; pyrrolidone carboxylic acid; 2-aminoadipic acid; 3-aminoadipic acid; beta-alanine; beta-aminoproprionic acid; 2-aminobutyric acid; 4-aminobutyric acid; piperidinic acid; 6-aminocaproic acid; 6-aminoheptanoic acid; 2-aminoheptanoic acid; 2-aminoisobutyric acid; 3-aminoisobutyric acid; 2-aminopimelic acid; 2,4 diaminobutyric acid; desmosine; 2,2 diaminopimelic acid; 2,3 diaminopropionic acid; N-ethylglycine; N-ethylasparagine; hydroxylysine; allo-hydroxylysine; 3-hydroxyproline; 4-hydroxyproline; isodesmosine; allo-isoleucine; N-methylglycine; sarcosine; methylisoleucine; methyllysine; methylvaline; norvaline; norleucine; 6-aminohexanoic acid; citrulline; cysteic acid; cyclohexylalanine; alpha-amino isobutyric acid; t-butylglycine; t-butylalanine; phenylglycine; an N-alpha-methyl amino acid, a C-alpha-methyl amino acid, a beta-methyl amino acid, or orthinine.  
     
     
         25 . The method of  claim 21 , wherein said second protein is a viral protein.  
     
     
         26 . A method of inhibiting a formation of a metastasis of a fibrosarcoma or Kaposi's sarcoma in a subject, comprising administering to said subject a compound that inhibits an interaction of a first protein and an immunoreceptor tyrosine-based activation motif (ITAM) of a second protein, thereby inhibiting a formation of a metastasis of a fibrosarcoma or Kaposi's sarcoma in a subject.  
     
     
         27 . The method of  claim 26 , wherein said compound comprises a peptide homologous to said ITAM.  
     
     
         28 . The method of  claim 27 , wherein said peptide comprises a modification selected from hydroxylation, amidation, esterification, formylation, gamma-carboxyglutamic acid hydroxylation, methylation, phosphorylation, sulfation, glycosylation, reduction, oxidation, disulfide modification, introduction of a thioether bond, introduction of a thiolester bond, a backbone condensation, or a biotinylation.  
     
     
         29 . The method of  claim 27 , wherein said peptide comprises an amino acid selected from a D amino acid; pyrrolidone carboxylic acid; 2-aminoadipic acid; 3-aminoadipic acid; beta-alanine; beta-aminoproprionic acid; 2-aminobutyric acid; 4-aminobutyric acid; piperidinic acid; 6-aminocaproic acid; 6-aminoheptanoic acid; 2-aminoheptanoic acid; 2-aminoisobutyric acid; 3-aminoisobutyric acid; 2-aminopimelic acid; 2,4 diaminobutyric acid; desmosine; 2,2 diaminopimelic acid; 2,3 diaminopropionic acid; N-ethylglycine; N-ethylasparagine; hydroxylysine; allo-hydroxylysine; 3-hydroxyproline; 4-hydroxyproline; isodesmosine; allo-isoleucine; N-methylglycine; sarcosine; methylisoleucine; methyllysine; methylvaline; norvaline; norleucine; 6-aminohexanoic acid; citrulline; cysteic acid; cyclohexylalanine; alpha-amino isobutyric acid; t-butylglycine; t-butylalanine; phenylglycine; an N-alpha-methyl amino acid, a C-alpha-methyl amino acid, a beta-methyl amino acid, or orthinine.  
     
     
         30 . The method of  claim 26 , wherein said second protein is a viral protein.  
     
     
         31 . A method of treating a Burkitt's lymphoma, a Hodgkin's disease, or a nasopharyngeal carcinoma (NPC) in a subject, comprising administering to said subject a compound that inhibits an interaction of a first protein and an immunoreceptor tyrosine-based activation motif (ITAM) of a second protein, thereby treating a Burkitt's lymphoma, a Hodgkin's disease, or a NPC in a subject.  
     
     
         32 . The method of  claim 31 , wherein said compound comprises a peptide homologous to said ITAM.  
     
     
         33 . The method of  claim 32 , wherein said peptide comprises a modification selected from hydroxylation, amidation, esterification, formylation, gamma-carboxyglutamic acid hydroxylation, methylation, phosphorylation, sulfation, glycosylation, reduction, oxidation, disulfide modification, introduction of a thioether bond, introduction of a thiolester bond, a backbone condensation, or a biotinylation.  
     
     
         34 . The method of  claim 32 , wherein said peptide comprises an amino acid selected from a D amino acid; pyrrolidone carboxylic acid; 2-aminoadipic acid; 3-aminoadipic acid; beta-alanine; beta-aminoproprionic acid; 2-aminobutyric acid; 4-aminobutyric acid; piperidinic acid; 6-aminocaproic acid; 6-aminoheptanoic acid; 2-aminoheptanoic acid; 2-aminoisobutyric acid; 3-aminoisobutyric acid; 2-aminopimelic acid; 2,4 diaminobutyric acid; desmosine; 2,2 diaminopimelic acid; 2,3 diaminopropionic acid; N-ethylglycine; N-ethylasparagine; hydroxylysine; allo-hydroxylysine; 3-hydroxyproline; 4-hydroxyproline; isodesmosine; allo-isoleucine; N-methylglycine; sarcosine; methylisoleucine; methyllysine; methylvaline; norvaline; norleucine; 6-aminohexanoic acid; citrulline; cysteic acid; cyclohexylalanine; alpha-amino isobutyric acid; t-butylglycine; t-butylalanine; phenylglycine; an N-alpha-methyl amino acid, a C-alpha-methyl amino acid, a beta-methyl amino acid, or orthinine.  
     
     
         35 . The method of  claim 31 , wherein said second protein is a viral protein.  
     
     
         36 . A method of reducing an incidence of a Burkitt's lymphoma, a Hodgkin's disease, or a nasopharyngeal carcinoma (NPC) in a subject, comprising administering to said subject a compound that inhibits an interaction of a first protein and an immunoreceptor tyrosine-based activation motif (ITAM) of a second protein, thereby reducing an incidence of a Burkitt's lymphoma, a Hodgkin's disease, or NPC in a subject.  
     
     
         37 . The method of  claim 36 , wherein said compound comprises a peptide homologous to said ITAM.  
     
     
         38 . The method of  claim 37 , wherein said peptide comprises a modification selected from hydroxylation, amidation, esterification, formylation, gamma-carboxyglutamic acid hydroxylation, methylation, phosphorylation, sulfation, glycosylation, reduction, oxidation, disulfide modification, introduction of a thioether bond, introduction of a thiolester bond, a backbone condensation, or a biotinylation.  
     
     
         39 . The method of  claim 37 , wherein said peptide comprises an amino acid selected from a D amino acid; pyrrolidone carboxylic acid; 2-aminoadipic acid; 3-aminoadipic acid; beta-alanine; beta-aminoproprionic acid; 2-aminobutyric acid; 4-aminobutyric acid; piperidinic acid; 6-aminocaproic acid; 6-aminoheptanoic acid; 2-aminoheptanoic acid; 2-aminoisobutyric acid; 3-aminoisobutyric acid; 2-aminopimelic acid; 2,4 diaminobutyric acid; desmosine; 2,2 diaminopimelic acid; 2,3 diaminopropionic acid; N-ethylglycine; N-ethylasparagine; hydroxylysine; allo-hydroxylysine; 3-hydroxyproline; 4-hydroxyproline; isodesmosine; allo-isoleucine; N-methylglycine; sarcosine; methylisoleucine; methyllysine; methylvaline; norvaline; nor-leucine; 6-aminohexanoic acid; citrulline; cysteic acid; cyclohexylalanine; alpha-amino isobutyric acid; t-butylglycine; t-butylalanine; phenylglycine; an N-alpha-methyl amino acid, a C-alpha-methyl amino acid, a beta-methyl amino acid, or orthinine.  
     
     
         40 . The method of  claim 36 , wherein said second protein is a viral protein.  
     
     
         41 . A method of inhibiting a formation of a metastasis of a Burkitt's lymphoma, a Hodgkin's disease, or a nasopharyngeal carcinoma (NPC) in a subject, comprising administering to said subject a compound that inhibits an interaction of a first protein and an immunoreceptor tyrosine-based activation motif (ITAM) of a second protein, thereby inhibiting a formation of a metastasis of a Burkitt's lymphoma, a Hodgkin's disease, or a NPC in a subject.  
     
     
         42 . The method of  claim 41 , wherein said compound comprises a peptide homologous to said ITAM.  
     
     
         43 . The method of  claim 42 , wherein said peptide comprises a modification selected from hydroxylation, amidation, esterification, formylation, gamma-carboxyglutamic acid hydroxylation, methylation, phosphorylation, sulfation, glycosylation, reduction, oxidation, disulfide modification, introduction of a thioether bond, introduction of a thiolester bond, a backbone condensation, or a biotinylation.  
     
     
         44 . The method of  claim 42 , wherein said peptide comprises an amino acid selected from a D amino acid; pyrrolidone carboxylic acid; 2-aminoadipic acid; 3-aminoadipic acid; beta-alanine; beta-aminoproprionic acid; 2-aminobutyric acid; 4-aminobutyric acid; piperidinic acid; 6-aminocaproic acid; 6-aminoheptanoic acid; 2-aminoheptanoic acid; 2-aminoisobutyric acid; 3-aminoisobutyric acid; 2-aminopimelic acid; 2,4 diaminobutyric acid; desmosine; 2,2 diaminopimelic acid; 2,3 diaminopropionic acid; N-ethylglycine; N-ethylasparagine; hydroxylysine; allo-hydroxylysine; 3-hydroxyproline; 4-hydroxyproline; isodesmosine; allo-isoleucine; N-methylglycine; sarcosine; methylisoleucine; methyllysine; methylvaline; norvaline; norleucine; 6-aminohexanoic acid; citrulline; cysteic acid; cyclohexylalanine; alpha-amino isobutyric acid; t-butylglycine; t-butylalanine; phenylglycine; an N-alpha-methyl amino acid, a C-alpha-methyl amino acid, a beta-methyl amino acid, or orthinine.  
     
     
         45 . The method of  claim 41 , wherein said second protein is a viral protein.  
     
     
         46 . A method of treating a B cell proliferative disorder in a subject, comprising administering to said subject a compound that inhibits an interaction of a first protein and an immunoreceptor tyrosine-based activation motif (ITAM) of a second protein, thereby treating a B cell proliferative disorder in a subject.  
     
     
         47 . A method of reducing an incidence of a B cell proliferative disorder in a subject, comprising administering to said subject a compound that inhibits an interaction of a first protein and an immunoreceptor tyrosine-based activation motif (ITAM) of a second protein, thereby reducing an incidence of a B cell proliferative disorder in a subject.  
     
     
         48 . A method of treating a mast cell activation disorder in a subject, comprising administering to said subject a compound that inhibits an interaction of a first protein and an immunoreceptor tyrosine-based activation motif (ITAM) of a second protein, thereby treating a mast cell activation disorder in a subject.  
     
     
         49 . A method of reducing an incidence of a mast cell activation disorder in a subject, comprising administering to said subject a compound that inhibits an interaction of a first protein and an immunoreceptor tyrosine-based activation motif (ITAM) of a second protein, thereby reducing an incidence of a mast cell activation disorder in a subject.