US2014051646A1PendingUtilityA1

Cell penetrating peptides

Assignee: MEDICAL RES COUNCILPriority: Jun 4, 2008Filed: Aug 14, 2013Published: Feb 20, 2014
Est. expiryJun 4, 2028(~1.9 yrs left)· nominal 20-yr term from priority
A61P 7/06A61P 9/00A61P 35/00A61P 25/14A61P 25/16A61P 3/00A61P 25/28A61P 17/00A61P 21/00A61K 31/7125C07K 14/001C07K 2319/10A61P 21/04A61K 47/64C07K 7/02A61K 47/48246
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Claims

Abstract

The present invention discloses cell penetrating peptides and conjugates of a cell penetrating peptide and a cargo molecule.

Claims

exact text as granted — not AI-modified
1 . A method of treating, preventing or ameliorating a disease in a patient in need of such treatment, prevention or amelioration, the method comprising administering a peptide-cargo conjugate to the patient thereby treating, preventing or ameliorating the disease, wherein the peptide-cargo conjugate comprises a peptide chemically conjugated to a cargo molecule, the peptide having a primary sequence structure comprised of at least three domains, having the arrangement:
 N-terminus [Domain 1]-[Domain 2]-[Domain 3] C-terminus in which:   (i) Domain 1 comprises a sequence chosen from:   
       
         
           
                 
                 
                 
               
                     
                   RXRRBRRXR 
                   [SEQ ID NO: 81] 
                 
                     
                     
                 
                     
                   RBRRXRRBR, or 
                   [SEQ ID NO: 82] 
                 
                     
                     
                 
                     
                   RXRRXR 
                   [SEQ ID NO: 74] 
                 
             
                
                
                
                
                
               
            
           
         
         (ii) Domain 2 comprises a sequence chosen from: 
       
       
         
           
                 
                 
                 
               
                     
                   ILFQ, or 
                   [SEQ ID NO: 86] 
                 
                     
                     
                 
                     
                   ILIQ 
                   [SEQ ID NO: 202] 
                 
             
                
                
                
               
            
           
         
         (iii) Domain 3 comprises a sequence chosen from: 
       
       
         
           
                 
                 
                 
               
                     
                   RXRBRXR 
                   [SEQ ID NO: 159] 
                 
                     
                     
                 
                     
                   RBRXRBR 
                   [SEQ ID NO: 162] 
                 
                     
                     
                 
                     
                   RXRRXR 
                   [SEQ ID NO: 161] 
                 
                     
                     
                 
                     
                   RXRXRXR 
                   [SEQ ID NO: 160] 
                 
                     
                     
                 
                     
                   RXRBRX 
                   [SEQ ID NO: 163] 
                 
                     
                     
                 
                     
                   MKWHK, or 
                   [SEQ ID NO: 203] 
                 
                     
                     
                 
                     
                   WKWHK 
                   [SEQ ID NO: 204] 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       wherein X=aminohexanoyl, B=betaAlanine, 
       and wherein the cargo molecule comprises an active agent capable of treating, preventing or ameliorating the disease. 
     
     
         2 . The method of  claim 1 , wherein the cargo molecule is conjugated to the peptide at the C-terminus of the peptide. 
     
     
         3 . The method of  claim 1 , wherein the cargo molecule is chosen from a nucleic acid, peptide nucleic acid (PNA), phosphorodiamidate morpholino oligonucleotide (PMO), locked nucleic acid (LNA), antisense oligonucleotide, short interfering RNA (siRNA), peptide, cyclic peptide, protein, or drug. 
     
     
         4 . The method of  claim 1 , wherein the cargo molecule has a molecular weight less than 5,000 Da. 
     
     
         5 . The method of  claim 1 , wherein the disease is caused by a splicing deficiency. 
     
     
         6 . The method of  claim 5 , wherein the cargo molecule comprises an oligonucleotide, peptide nucleic acid (PNA), phosphorodiamidate morpholino oligonucleotide (PMO), or locked nucleic acid (LNA) capable of preventing or correcting the splicing defect and/or increasing the production of correctly spliced mRNA molecules. 
     
     
         7 . The method of  claim 1 , wherein the disease is chosen from Duchenne Muscular Dystrophy (DMD), Menkes Disease, β-thalassemia, frontotemporal dementia, parkinsonism, spinal muscular atrophy, Hutchinson-Gilford Progeria Syndrome, Ataxiatelangiectasia mutated (ATM), or cancer. 
     
     
         8 . The method of  claim 1 , wherein the cargo molecule is PNADMD [SEQ ID NO:115] or PMODMD [SEQ ID NO:206] or a molecule having at least 90% sequence identity to one of PNADMD [SEQ ID NO:115] or PMODMD [SEQ ID NO:206]. 
     
     
         9 . The method of  claim 1 , wherein the disease is Duchenne Muscular Dystrophy (DMD) and the cargo molecule is PNADMD [SEQ ID NO:115] or PMODMD [SEQ ID NO:206] or a molecule having at least 90% sequence identity to one of PNADMD [SEQ ID NO:115] or PMODMD [SEQ ID NO:206]. 
     
     
         10 . The method of  claim 1 , wherein Domain 2 comprises a sequence chosen from ILFQY [SEQ ID NO:88], or ILIQY [SEQ ID NO:296]. 
     
     
         11 . The method of  claim 1 , wherein Domain 2 comprises a sequence chosen from: 
       
         
           
                 
                 
                 
               
                     
                   Z 2 Z 3 ILFQZ 4 , or 
                   [SEQ ID NO: 89] 
                 
                     
                     
                 
                     
                   Z 2 Z 3 ILIQZ 4   
                   [SEQ ID NO: 297] 
                 
             
                
                
                
               
            
           
         
       
       wherein Z 2 =I or no amino acid, Z 3 =K, dK, H, R or no amino acid, Z 4 =N or Y. 
     
     
         12 . The method of  claim 1 , wherein Domain 3 comprises a sequence chosen from: 
       
         
           
                 
                 
                 
               
                     
                   Z 7 Z 8 MKWHK, or 
                   [SEQ ID NO: 298] 
                 
                     
                     
                 
                     
                   Z 7 Z 8 WKWHK 
                   [SEQ ID NO: 299] 
                 
             
                
                
                
               
            
           
         
       
       where Z 7 =R, dR, K or dK (wherein d=D-aminoacid)
 Z 8 =R, H or K. 
 
     
     
         13 . The method of  claim 1 , wherein Domain 3 comprises a sequence chosen from: 
       
         
           
                 
                 
                 
               
                     
                   dRRMKWHK 
                   [SEQ ID NO: 111] 
                 
                     
                     
                 
                     
                   dRHMKWHK 
                   [SEQ ID NO: 155] 
                 
                     
                     
                 
                     
                   dRRWKWHK 
                   [SEQ ID NO: 157] 
                 
                     
                     
                 
                     
                   dKRMKWHK, or 
                   [SEQ ID NO: 156] 
                 
                     
                     
                 
                     
                   dKHWKWHK. 
                   [SEQ ID NO: 158] 
                 
             
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         14 . The method of  claim 1 , wherein Domain 1 comprises a sequence chosen from: 
       
         
           
                 
                 
               
                   (RXR) n  where n = 2, 3 or 4, or 
                   [SEQ ID NOs: 74-76] 
                 
                     
                 
                   RXRRXRIdR. 
                   [SEQ ID NO: 80] 
                 
             
                
                
                
               
            
           
         
       
     
     
         15 . The method of  claim 1 , wherein the peptide has a maximum length of 30 residues, including natural amino acids, X and B residues. 
     
     
         16 . The method of  claim 1 , wherein Domain 1 has a length of from 6 to 11 residues, Domain 2 has a length of from 4 to 9 residues, and Domain 3 has a length of from 4 to 15 residues, wherein the lengths include natural amino acids, X and B residues. 
     
     
         17 . The method of  claim 1 , wherein the peptide comprises, or consists of, a sequence chosen from one of SEQ ID NOs:242-295 ( FIG. 25 ) or a sequence having at least 90% sequence identity to one of SEQ ID NOs:242-295 ( FIG. 25 ). 
     
     
         18 . The method of  claim 1 , wherein the peptide comprises, or consists of, the sequence of Domains 1 to 3 or Domains 1 to 4 of one of Pip-5e, Pip-5j or Pip-5l (SEQ ID NOs:230, 234 or 236) or a sequence having at least 90% sequence Identity to the sequence of Domains 1 to 3 or Domains 1 to 4 of one of Pip-5e, Pip-5j or Pip-5l (SEQ ID NOs:230, 234 or 236). 
     
     
         19 . The method of  claim 1 , wherein the peptide comprises, or consists of, a sequence chosen from one of: 
       
         
           
                 
                 
                 
               
                     
                   Z 1 Z 2 Z 3 ILFQZ 4 Z 5 RMKWHK 
                   [SEQ ID NO: 113] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILFQZ 4 Z 5 HMKWHK 
                   [SEQ ID NO: 183] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILFQZ 4 Z 5 RWKWHK 
                   [SEQ ID NO: 184] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILFQZ 4 Z 5 HWKWHK 
                   [SEQ ID NO: 185] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILFQZ 4 RXRBRXR 
                   [SEQ ID NO: 186] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILFQZ 4 RXRXRXR 
                   [SEQ ID NO: 187] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILFQZ 4 RXRRXR 
                   [SEQ ID NO: 188] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILFQZ 4 RBRXRBR 
                   [SEQ ID NO: 189] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILFQZ 4 RXRBRXR 
                   [SEQ ID NO: 190] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILFQZ 4 RXRBRX 
                   [SEQ ID NO: 191] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILIQZ 4 Z 5 RMKWHK 
                   [SEQ ID NO: 192] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILIQZ 4 Z 5 HMKWHK 
                   [SEQ ID NO: 193] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILIQZ 4 Z 5 RWKWHK 
                   [SEQ ID NO: 194] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILIQZ 4 Z 5 HWKWHK 
                   [SEQ ID NO: 195] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILIQZ 4 RXRBRXR 
                   [SEQ ID NO: 196] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILIQZ 4 RXRXRXR 
                   [SEQ ID NO: 197] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILIQZ 4 RXRRXR 
                   [SEQ ID NO: 198] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILIQZ 4 RBRXRBR 
                   [SEQ ID NO: 199] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILIQZ 4 RXRBRXR 
                   [SEQ ID NO: 200] 
                 
                     
                     
                 
                     
                   Z 1 Z 2 Z 3 ILIQZ 4 RXRBRX 
                   [SEQ ID NO: 201] 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       wherein:
 Z 1 =(RXFR) n  where n=2, 3, or 4; (RBR), where n=2, 3, or 4; (R), where m=5, 6, 7 or 8; RXRRXRIdR [SEQ ID NO:80]; RXRRBRRXR [SEQ ID NO:81]; or RBRRXRRBR SEQ ID NO:82; 
 Z 2 =I or no amino acid 
 Z 3 =K, dK, H, R or no amino acid 
 Z 4 =N, Y 
 Z 5 =R or dR. K or dK 
 
       wherein d=D-aminoacid. 
     
     
         20 . The method of  claim 1 , wherein the peptide comprises, or consists of, a sequence chosen from one of SEQ ID NOs:34-73 ( FIG. 16 ), or a sequence having at least 90% sequence identity to one of SEQ ID NOs:34-73 ( FIG. 16 ). 
     
     
         21 . The method of  claim 1 , wherein the peptide further comprises a linker sequence at the C-terminus. 
     
     
         22 . The method of  claim 21 , wherein the linker sequence is chosen from BCys, XCys, Cys, GGCys, BBCys, BXCys, XBCys, BX, or XB.

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