Methods for attenuating release of inflammatory mediators and peptides useful therein

Abstract

The present invention includes methods of inhibiting or suppressing cellular secretory processes. More specifically the present invention relates to inhibiting or reducing the release of inflammatory mediators from inflammatory cells by inhibiting the mechanism associated with the release of inflammatory mediators from granules in inflammatory cells. In this regard, the present invention discloses an intracellular signaling mechanism that illustrates several novel intracellular targets for pharmacological intervention in disorders involving secretion of inflammatory mediators from vesicles in inflammatory cells. Peptide fragments and variants thereof of MANS peptide as disclosed in the present invention are useful in such methods.

1 . A method of inhibiting the release of at least one inflammatory mediator from a granule in at least one inflammatory cell in a tissue and/or fluid of a subject comprising:
 administration to said tissue and/or fluid a therapeutically effective amount of a pharmaceutical composition comprising at least one peptide having an amino acid sequence selected from the group consisting of:   (a) an amino acid sequence having from 4 to 23 contiguous amino acids of a reference sequence, GAQFSKTAAKGEAAAERPGEAAVA (SEQ ID NO. 1);   (b) an amino acid sequence having the sequence, GAQFSKTAAKGEAAAERPGEAAVA (SEQ ID NO. 1); and   (c) an amino acid sequence substantially identical to the sequence defined in (a),   wherein the C-terminal amino acid of the peptide is optionally independently chemically modified, and the N-terminal amino acid of the peptide is independently chemically modified by acylation with a carboxylic acid selected from the group consisting of a C2 to C13 saturated or unsaturated aliphatic carboxylic acid, a C14 saturated or unsaturated aliphatic carboxylic acid, a C15 to C24 saturated or unsaturated aliphatic carboxylic acid, and trifluoroacetic acid, or is not chemically modified, with the proviso that said peptide is modified by acylation when its amino acid sequence begins with the sequence GAQF of the reference sequence by acylation only with a carboxylic acid selected from the group consisting of a C2 to C13 saturated or unsaturated aliphatic carboxylic acid, a C14 unsaturated aliphatic carboxylic acid, a C15 to C24 saturated or unsaturated aliphatic carboxylic acid, and trifluoroacetic acid, or is not chemically modified, wherein said peptide, optionally combined with a pharmaceutically acceptable carrier, and in a therapeutically effective inflammatory mediator release-reducing amount to reduce the release of said inflammatory mediator from at least one inflammatory cell as compared to release of said inflammatory mediator from at least one of the same type of inflammatory cell that would occur in the absence of said at least one peptide.   
     
     
         2 .- 21 . (canceled) 
     
     
         22 . The method according to  claim 1 , wherein said subject is afflicted by a respiratory disease. 
     
     
         23 . The method according to  claim 22 , wherein said respiratory disease is selected from the group consisting of asthma, chronic bronchitis, COPD and cystic fibrosis. 
     
     
         24 . The method according to  claim 1 , wherein said subject is a mammal. 
     
     
         25 . The method according to  claim 24 , wherein said mammal is selected from the group consisting of humans, canines, equines and felines. 
     
     
         26 . The method according to  claim 1 , wherein said administration is selected from the group consisting of topical administration, parenteral administration, rectal administration, pulmonary administration, nasal administration, and oral administration. 
     
     
         27 . The method according to  claim 26 , wherein said pulmonary administration comprises an aerosol. 
     
     
         28 . The method according to  claim 27 , wherein said aerosol is generated from a dry powder inhaler, a metered dose inhaler or nebulizer. 
     
     
         29 . The method according to  claim 1 , further comprising administration to said subject of a second molecule selected from the group consisting of an antibiotic, an antiviral compound, an antiparasitic compound, an anti-inflammatory compound, and an immunomodulator. 
     
     
         30 .- 37 . (canceled) 
     
     
         38 . An isolated peptide having an amino acid sequence selected from the group consisting of:
 (a) an amino acid sequence having from 4 to 23 contiguous amino acids of a reference sequence, GAQFSKTAAKGEAAAERPGEAAVA (SEQ ID NO. 1);   (b) an amino acid sequence having the sequence, GAQFSKTAAKGEAAAERPGEAAVA (SEQ ID NO. 1); and   (c) an amino acid sequence substantially identical to the sequence defined in (a),   wherein the C-terminal amino acid of the peptide is optionally independently chemically modified, and the N-terminal amino acid of the peptide is independently chemically modified by acylation with a carboxylic acid selected from the group consisting of a C2 to C13 saturated or unsaturated aliphatic carboxylic acid, a C14 saturated or unsaturated aliphatic carboxylic acid, a C15 to C24 saturated or unsaturated aliphatic carboxylic acid, and trifluoroacetic acid, or is not chemically modified, with the proviso that said peptide is modified by acylation when its amino acid sequence begins with the sequence GAQF of the reference sequence by acylation only with a carboxylic acid selected from the group consisting of a C2 to C13 saturated or unsaturated aliphatic carboxylic acid, a C14 unsaturated aliphatic carboxylic acid, a C15 to C24 saturated or unsaturated aliphatic carboxylic acid, and trifluoroacetic acid, or is not chemically modified, wherein said peptide, optionally combined with a pharmaceutically acceptable carrier, and in a therapeutically effective inflammatory mediator release-reducing amount to reduce the release of said inflammatory mediator from at least one inflammatory cell as compared to release of said inflammatory mediator from at least one of the same type of inflammatory cell that would occur in the absence of said at least one peptide.   
     
     
         39 . The peptide according to  claim 38 , wherein said peptide is acetylated at the alpha N-terminal amino acid. 
     
     
         40 . (canceled) 
     
     
         41 . The peptide according to  claim 40 , wherein said peptide consists of acetyl-peptide 106 (SEQ ID NO: 106). 
     
     
         42 . The method according to  claim 38 , wherein said peptide consists of at least four contiguous amino acid residues. 
     
     
         43 . (canceled) 
     
     
         44 . The peptide according to  claim 38 , wherein said peptide is myristoylated at the alpha N-terminal amino acid. 
     
     
         45 . The peptide according to  claim 38 , wherein said peptide is amidated with ammonia at the alpha C-terminal amino acid. 
     
     
         46 . The peptide according to  claim 38 , wherein the peptide comprises an amino acid sequence of (a) wherein the N-terminal amino acid of the amino acid sequence of (a) is selected from amino acid position 2 to 21 of the reference sequence, GAQFSKTAAKGEAAAERPGEAAVA (SEQ ID NO. 1). 
     
     
         47 .- 53 . (canceled) 
     
     
         54 . A pharmaceutical composition comprising an isolated peptide according to  claim 38  and a pharmaceutically acceptable carrier. 
     
     
         55 . The pharmaceutical composition according to  claim 54 , wherein said composition is sterile, sterilizable or sterilized. 
     
     
         56 . A kit comprising at least one isolated peptide according to  claim 38 . 
     
     
         57 . (canceled) 
     
     
         58 . A method of treating or preventing adult respiratory distress syndrome (ARDS), acute pulmonary inflammation, or acute thermal injury in a subject in need thereof, comprising:
 administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one peptide having an amino acid sequence selected from the group consisting of:   (a) an amino acid sequence having from 4 to 23 contiguous amino acids of a reference sequence, GAQFSKTAAKGEAAAERPGEAAVA (SEQ ID NO. 1);   (b) an amino acid sequence having the sequence, GAQFSKTAAKGEAAAERPGEAAVA (SEQ ID NO. 1); and   (c) an amino acid sequence substantially identical to the sequence defined in (a),   wherein the C-terminal amino acid of the peptide is optionally independently chemically modified, and the N-terminal amino acid of the peptide is independently chemically modified by acylation with a carboxylic acid selected from the group consisting of a C2 to C13 saturated or unsaturated aliphatic carboxylic acid, a C14 saturated or unsaturated aliphatic carboxylic acid, a C15 to C24 saturated or unsaturated aliphatic carboxylic acid, and trifluoroacetic acid, or is not chemically modified, with the proviso that said peptide is modified by acylation when its amino acid sequence begins with the sequence GAQF of the reference sequence by acylation only with a carboxylic acid selected from the group consisting of a C2 to C13 saturated or unsaturated aliphatic carboxylic acid, a C14 unsaturated aliphatic carboxylic acid, a C15 to C24 saturated or unsaturated aliphatic carboxylic acid, and trifluoroacetic acid, or is not chemically modified.