US2017190748A1PendingUtilityA1

Anti-inflammatory proteins and peptides and methods of preparation and use thereof

Assignee: MANUKAMED HOLDINGS LTD PARTNERSHIPPriority: Jun 22, 2012Filed: Jan 11, 2017Published: Jul 6, 2017
Est. expiryJun 22, 2032(~5.9 yrs left)· nominal 20-yr term from priority
A61P 9/00A61P 3/10A61P 39/06A61P 37/02A61P 37/08A61P 37/06A61P 31/00A61P 31/12A61P 29/00A61P 27/02A61P 25/28A61P 31/22A61P 35/00A61P 11/06A61P 17/02A61P 15/00A61P 11/00A61P 19/02A61P 1/04A61P 17/06A61P 25/00A61P 17/00A61K 38/00C12Q 1/37G01N 2333/43565C07K 14/8139C07K 5/1019C07K 5/0808A61K 38/1767A61K 35/644C07K 5/1013A61L 2300/41G01N 2333/96466A61L 2300/252C07K 5/0812A61L 15/32G01N 2500/04G01N 2500/20C07K 14/43572C07K 5/1024C07K 5/0815A61L 15/44A61L 2300/25C12Y 304/22001
28
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Claims

Abstract

The present disclose relates to anti-inflammatory proteins/peptides, their uses, methods of preparation and methods of their detection. In particular, the invention relates to major royal jelly proteins modified by methyglyoxal and fragments thereof from a Leptospermum derived honey and royal jelly.

Claims

exact text as granted — not AI-modified
1 .- 24 . (canceled) 
     
     
         25 . An isolated functional fragment of a major royal jelly protein (MRJP), which has anti-inflammatory capacity, and which is not a naturally occurring functional fragment of a major royal jelly protein, wherein a lysine or arginine amino acid residue of the functional fragment has been chemically modified by methylglyoxal (MGO), and wherein:
 a. the functional fragment comprises an amino acid sequence selected from the group consisting of:   
       
         
           
                 
                 
               
                     
                   i.  
                 
                     
                   (SEQ ID NO: 11) 
                 
                     
                   KNNNQNDN; 
                 
                     
                     
                 
                     
                   ii.  
                 
                     
                   (SEQ ID NO: 12) 
                 
                     
                   KLH; 
                 
                     
                     
                 
                     
                   iii.  
                 
                     
                   (SEQ ID NO: 13) 
                 
                     
                   KSNNRHNNND; 
                 
                     
                     
                 
                     
                   iv.  
                 
                     
                   (SEQ ID NO: 14) 
                 
                     
                   KHNN; 
                 
                     
                     
                 
                     
                   v.  
                 
                     
                   (SEQ ID NO: 15) 
                 
                     
                   KNQAHLD; 
                 
                     
                     
                 
                     
                   vi.  
                 
                     
                   (SEQ ID NO: 16) 
                 
                     
                   KNTRCISP; 
                 
                     
                     
                 
                     
                   vii.  
                 
                     
                   (SEQ ID NO: 17) 
                 
                     
                   KTNFFSIFL;  
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       
         
           
                 
                 
               
                     
                   viii.  
                 
                     
                   (SEQ ID NO: 64) 
                 
                     
                     PFKISIHL ; 
                 
                     
                     
                 
                     
                   ix.  
                 
                     
                   (SEQ ID NO: 69) 
                 
                     
                     NQKNNNQNDN ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 66) 
                 
                     
                     NQKN ; 
                 
                     
                     
                 
                     
                   x.  
                 
                     
                   (SEQ ID NO: 71) 
                 
                     
                     SKLH ; 
                 
                     
                     
                 
                     
                   xi.  
                 
                     
                   (SEQ ID NO: 81) 
                 
                     
                     QNKHNN ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 80) 
                 
                     
                     KH ;  
                 
                     
                     
                 
                     
                   xii.  
                 
                     
                   (SEQ ID NO: 84) 
                 
                     
                     LVK ;  
                 
                     
                     
                 
                     
                   xiii.  
                 
                     
                   (SEQ ID NO: 86) 
                 
                     
                     LIR ;  
                 
                     
                     
                 
                     
                   xiv.  
                 
                     
                   (SEQ ID NO: 127) 
                 
                     
                     FDR ;  
                 
                     
                     
                 
                     
                   xv.  
                 
                     
                   (SEQ ID NO: 128) 
                 
                     
                     HNIR ;  
                 
                     
                   and 
                 
                     
                     
                 
                     
                   xvi.  
                 
                     
                   (SEQ ID NO: 130) 
                 
                     
                     FTK   
                 
                     
                     
                 
                     
                   (SEQ ID NO: 137) 
                 
                     
                     QNGNK ; 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       or
 b. the functional fragment comprises a peptide analogue selected from the group consisting of: 
 
       
         
           
                 
                 
               
                     
                   i. 
                 
                     
                   (SEQ ID NO: 24) 
                 
                     
                     Ac-NQKNNNQNDN-OH ; 
                 
                     
                     
                 
                     
                   ii. 
                 
                     
                   (SEQ ID NO: 27) 
                 
                     
                     Ac-NQKN-OH ; 
                 
                     
                     
                 
                     
                   iii. 
                 
                     
                   (SEQ ID NO: 31) 
                 
                     
                     Ac-SKLH-OH ; 
                 
                     
                     
                 
                     
                   iv. 
                 
                     
                   (SEQ ID NO: 43) 
                 
                     
                     Ac-QNKHNN-OH ;  
                 
                     
                     
                 
                     
                   (SEQ ID NO: 45) 
                 
                     
                     Ac-KH-OH ; 
                 
                     
                     
                 
                     
                   v. 
                 
                     
                   (SEQ ID NO: 48) 
                 
                     
                     Ac-LVK-OH ; 
                 
                     
                     
                 
                     
                   vi. 
                 
                     
                   (SEQ ID NO: 51) 
                 
                     
                   A c-LIR-OH ; 
                 
                     
                     
                 
                     
                   vii. 
                 
                     
                   (SEQ ID NO: 53) 
                 
                     
                     Ac-FDR-OH ; 
                 
                     
                     
                 
                     
                   viii. 
                 
                     
                   (SEQ ID NO: 59) 
                 
                     
                     Ac-HNIR-OH ; 
                 
                     
                     
                 
                     
                   ix. 
                 
                     
                   (SEQ ID NO: 88) 
                 
                     
                     H-FTK-OH ;  
                 
                     
                   and 
                 
                     
                     
                 
                     
                   x. 
                 
                     
                   (SEQ ID NO: 106) 
                 
                     
                     Ac-QNGNK-OH . 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         26 .- 29 . (canceled) 
     
     
         30 . The isolated functional fragment of  claim 25 , which is chemically synthesized. 
     
     
         31 . The isolated functional fragment of  claim 25 , which is recombinantly produced. 
     
     
         32 . A composition comprising the isolated functional fragment of  claim 25 , or an analogue thereof. 
     
     
         33 . A wound dressing comprising a functional fragment of a major royal jelly protein (MRJP); wherein the functional fragment has anti-inflammatory activity; wherein the functional fragment has been isolated, enriched, chemically synthesized, or recombinantly produced; wherein a lysine or arginine amino acid residue of the functional fragment has been chemically modified by methylglyoxal (MGO); and wherein:
 a. the functional fragment comprises an amino acid sequence selected from the group consisting of:   
       
         
           
                 
                 
               
                     
                   i. 
                 
                     
                   (SEQ ID NO: 11) 
                 
                     
                   KNNNQNDN; 
                 
                     
                     
                 
                     
                   ii. 
                 
                     
                   (SEQ ID NO: 12) 
                 
                     
                   KLH; 
                 
                     
                     
                 
                     
                   iii. 
                 
                     
                   (SEQ ID NO: 13) 
                 
                     
                   KSNNRHNNND; 
                 
                     
                     
                 
                     
                   iv. 
                 
                     
                   (SEQ ID NO: 14) 
                 
                     
                   KHNN; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 15) 
                 
                     
                   KNQAHLD; 
                 
                     
                     
                 
                     
                   vi.  
                 
                     
                   (SEQ ID NO: 16) 
                 
                     
                   KNTRCISP; 
                 
                     
                     
                 
                     
                   vii.  
                 
                     
                   (SEQ ID NO: 17) 
                 
                     
                   KTNFFSIFL; 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       
         
           
                 
                 
               
                     
                   viii. 
                 
                     
                   (SEQ ID NO: 64) 
                 
                     
                     PFKISIHL ; 
                 
                     
                     
                 
                     
                   ix. 
                 
                     
                   (SEQ ID NO: 69) 
                 
                     
                     NQKNNNQNDN ;  
                 
                     
                     
                 
                     
                   (SEQ ID NO: 66) 
                 
                     
                     NQKN ; 
                 
                     
                     
                 
                     
                   x. 
                 
                     
                   (SEQ ID NO: 71) 
                 
                     
                     SKLH ; 
                 
                     
                     
                 
                     
                   xi. 
                 
                     
                   (SEQ ID NO: 81) 
                 
                     
                     QNKHNN ;  
                 
                     
                     
                 
                     
                   (SEQ ID NO: 80) 
                 
                     
                     KH ; 
                 
                     
                     
                 
                     
                   xii. 
                 
                     
                   (SEQ ID NO: 84) 
                 
                     
                     LVK ; 
                 
                     
                     
                 
                     
                   xiii. 
                 
                     
                   (SEQ ID NO: 86) 
                 
                     
                     LIR ; 
                 
                     
                     
                 
                     
                   xiv. 
                 
                     
                   (SEQ ID NO: 127) 
                 
                     
                     FDR ; 
                 
                     
                     
                 
                     
                   xv. 
                 
                     
                   (SEQ ID NO: 128) 
                 
                     
                     HNIR ;  
                 
                     
                   and 
                 
                     
                     
                 
                     
                   xvi. 
                 
                     
                   (SEQ ID NO: 130) 
                 
                     
                   
                     FTK  
                   
                 
                     
                     
                 
                     
                   (SEQ ID NO: 137) 
                 
                     
                     QNGNK ; 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       or
 b. the functional fragment comprises a peptide analogue selected from the group consisting of: 
 
       
         
           
                 
                 
               
                     
                   i. 
                 
                     
                   (SEQ ID NO: 24) 
                 
                     
                     Ac-NQKNNNQNDN-OH ; 
                 
                     
                     
                 
                     
                   ii. 
                 
                     
                   (SEQ ID NO: 27) 
                 
                     
                     Ac-NQKN-OH ; 
                 
                     
                     
                 
                     
                   iii. 
                 
                     
                   (SEQ ID NO: 31) 
                 
                     
                     Ac-SKLH-OH ; 
                 
                     
                     
                 
                     
                   iv. 
                 
                     
                   (SEQ ID NO: 43) 
                 
                     
                     Ac-QNKHNN-OH ;  
                 
                     
                     
                 
                     
                   (SEQ ID NO: 45) 
                 
                     
                     Ac-KH-OH ; 
                 
                     
                     
                 
                     
                   v. 
                 
                     
                   (SEQ ID NO: 48) 
                 
                     
                     Ac-LVK-OH ; 
                 
                     
                     
                 
                     
                   vi. 
                 
                     
                   (SEQ ID NO: 51) 
                 
                     
                     Ac-LIR-OH ; 
                 
                     
                     
                 
                     
                   vii. 
                 
                     
                   (SEQ ID NO: 53) 
                 
                     
                     Ac-FDR-OH ; 
                 
                     
                     
                 
                     
                   viii. 
                 
                     
                   (SEQ ID NO: 59) 
                 
                     
                     Ac-HNIR-OH ; 
                 
                     
                     
                 
                     
                   ix. 
                 
                     
                   (SEQ ID NO: 88) 
                 
                     
                     H-FTK-OH ;  
                 
                     
                   and 
                 
                     
                     
                 
                     
                   x. 
                 
                     
                   (SEQ ID NO: 106) 
                 
                     
                     Ac-QNGNK-OH . 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         34 .- 44 . (canceled) 
     
     
         45 . A method of inhibiting Cathepsin B activity in a cellular tissue, comprising the step of contacting the cellular tissue with a functional fragment of a major royal jelly protein (MRJP); wherein the functional fragment has been isolated, enriched, synthesized, or recombinantly produced; wherein a lysine or arginine amino acid residue of the functional fragment has been chemically modified by methylglyoxal (MGO); and wherein
 a. the functional fragment comprises an amino acid sequence selected from the group consisting of:   
       
         
           
                 
                 
               
                     
                   i. 
                 
                     
                   (SEQ ID NO: 10) 
                 
                     
                     KISIHL ; 
                 
                     
                     
                 
                     
                   ii. 
                 
                     
                   (SEQ ID NO: 11) 
                 
                     
                     KNNNQNDN ; 
                 
                     
                     
                 
                     
                   iii. 
                 
                     
                   (SEQ ID NO: 12) 
                 
                     
                     KLH ; 
                 
                     
                     
                 
                     
                   iv. 
                 
                     
                   (SEQ ID NO: 13) 
                 
                     
                     KSNNRHNNND ; 
                 
                     
                     
                 
                     
                   v. 
                 
                     
                   (SEQ ID NO: 14) 
                 
                     
                     KHNN ; 
                 
                     
                     
                 
                     
                   vi. 
                 
                     
                   (SEQ ID NO: 15) 
                 
                     
                     KNQAHLD ; 
                 
                     
                     
                 
                     
                   vii. 
                 
                     
                   (SEQ ID NO: 16) 
                 
                     
                     KNTRCISP ; 
                 
                     
                     
                 
                     
                   viii. 
                 
                     
                   (SEQ ID NO: 17) 
                 
                     
                     KTNFFSIFL ; 
                 
                     
                     
                 
                     
                   ix. 
                 
                     
                   (SEQ ID NO: 64) 
                 
                     
                     PFKISIHL ; 
                 
                     
                     
                 
                     
                   x. 
                 
                     
                   (SEQ ID NO: 69) 
                 
                     
                     NQKNNNQNDN ;  
                 
                     
                     
                 
                     
                   (SEQ ID NO: 66) 
                 
                     
                     NQKN ; 
                 
                     
                     
                 
                     
                   xi. 
                 
                     
                   (SEQ ID NO: 71) 
                 
                     
                     SKLH ; 
                 
                     
                     
                 
                     
                   xii. 
                 
                     
                   (SEQ ID NO: 81) 
                 
                     
                     QNKHNN ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 80) 
                 
                     
                     KH ; 
                 
                     
                     
                 
                     
                   xiii. 
                 
                     
                   (SEQ ID NO: 84) 
                 
                     
                     LVK ; 
                 
                     
                     
                 
                     
                   xiv. 
                 
                     
                   (SEQ ID NO: 86) 
                 
                     
                     LIR ; 
                 
                     
                     
                 
                     
                   xv. 
                 
                     
                   (SEQ ID NO: 127) 
                 
                     
                     FDR ; 
                 
                     
                     
                 
                     
                   xvi. 
                 
                     
                   (SEQ ID NO: 128) 
                 
                     
                     HNIR ;  
                 
                     
                   and 
                 
                     
                     
                 
                     
                   xvii.  
                 
                     
                   (SEQ ID NO: 130) 
                 
                     
                   
                     FTK  
                   
                 
                     
                     
                 
                     
                   (SEQ ID NO: 137) 
                 
                     
                     QNGNK ; 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       or
 b. the functional fragment comprises a peptide analogue selected from the group consisting of: 
 
       
         
           
                 
                 
               
                     
                   i. 
                 
                     
                   (SEQ ID NO: 24) 
                 
                     
                     Ac-NQKNNNQNDN-OH ; 
                 
                     
                     
                 
                     
                   ii. 
                 
                     
                   (SEQ ID NO: 27) 
                 
                     
                     Ac-NQKN-OH ; 
                 
                     
                     
                 
                     
                   iii. 
                 
                     
                   (SEQ ID NO: 31) 
                 
                     
                     Ac-SKLH-OH ; 
                 
                     
                     
                 
                     
                   iv. 
                 
                     
                   (SEQ ID NO: 43) 
                 
                     
                     Ac-QNKHNN-OH ;  
                 
                     
                     
                 
                     
                   (SEQ ID NO: 45) 
                 
                     
                     Ac-KH-OH ; 
                 
                     
                     
                 
                     
                   v. 
                 
                     
                   (SEQ ID NO: 48) 
                 
                     
                     Ac-LVK-OH ; 
                 
                     
                     
                 
                     
                   vi. 
                 
                     
                   (SEQ ID NO: 51) 
                 
                     
                     Ac-LIR-OH ; 
                 
                     
                     
                 
                     
                   vii. 
                 
                     
                   (SEQ ID NO: 53) 
                 
                     
                     Ac-FDR-OH ; 
                 
                     
                     
                 
                     
                   viii. 
                 
                     
                   (SEQ ID NO: 59) 
                 
                     
                     Ac-HNIR-OH ; 
                 
                     
                     
                 
                     
                   ix. 
                 
                     
                   (SEQ ID NO: 88) 
                 
                     
                     H-FTK-OH ; 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   x. 
                 
                     
                   (SEQ ID NO: 106) 
                 
                     
                     Ac-QNGNK-OH . 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         46 .- 53 . (canceled) 
     
     
         54 . The method of  claim 45 , wherein the method reduces inflammation in the cellular tissue. 
     
     
         55 . The method of  claim 54 , wherein the inflammation is associated with one or more of the group consisting of: an inflammatory disorder, a cardiovascular disorder, a neurological disorder, a pulmonary disorder, a proliferative disorder, an infectious disease or associated syndrome, an allergic, immunological or autoimmune disorder, and inflammation associated with a wound. 
     
     
         56 . The method of  claim 54 , wherein the inflammation is associated with one or more of the group consisting of: rheumatoid arthritis and polyarthritis. 
     
     
         57 . The method of  claim 54 , wherein the inflammation is associated with one or more of the group consisting of: Alzheimer's disease, progressive multifocal leukoencephalopathy (PML), and multiple sclerosis. 
     
     
         58 . The method of  claim 54 , wherein the inflammation is associated with one or more of the group consisting of: asthma, bronchitis, bronchopneumonia, adult respiratory distress syndrome, emphysema, and chronic obstructive pulmonary disease (COPD). 
     
     
         59 . The method of  claim 54 , wherein the inflammation is associated with one or more of the group consisting of: genital herpes, Epstein-Barr virus (EBV) infection, EBV associated encephalitis, and cytomegalovirus (CMV) associated choreoretinitis. 
     
     
         60 . The method of  claim 54 , wherein the inflammation is associated with one or more of the group consisting of: Wegeneres granulomatosis, choreoretinitis, encephalitis, gastroenteritis, uveitis, psoriasis, and dermatitis. 
     
     
         61 . The method of  claim 54 , wherein the inflammation is associated with melanoma. 
     
     
         62 . The method of  claim 55 , wherein the wound is one or more of the group consisting of: a diabetic ulcer, fungating wound, puncture wound, cut, bite, and surgical wound. 
     
     
         63 . The method of  claim 54 , wherein the method comprises administration of the functional fragment to a subject, and wherein the administration includes one or more of oral, topical, and parenteral administration. 
     
     
         64 . The method of  claim 54 , wherein the method comprises administration of the functional fragment to a subject, and wherein the administration includes one or more of intravenous, intravitreal, or intramuscular administration. 
     
     
         65 . The method of  claim 54 , wherein the method reduces the rate of phagocytosis by immune system cells. 
     
     
         66 . The method of  claim 54 , wherein the method inhibits receptors for phagocytosis on immune system cells. 
     
     
         67 . The method of  claim 54 , wherein the method reduces respiratory burst and release of reactive oxygen species from inflammatory cells.

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