Stapled intracellular-targeting antimicrobial peptides to treat infection

Abstract

Structurally stabilized, e.g., stapled, peptides with the ability to translocate through microbial cell membranes to the interior of microbial cells and exert a biological activity there are provided, as are methods of designing, making and using such peptides.

What is claimed is: 
     
         1 . A compound having the formula: 
       
         
           
           
               
               
           
         
         or a pharmaceutically acceptable salt thereof,
 wherein; 
 each R 1  and R 2  is independently H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkylalkyl, heteroarylalkyl, or heterocyclylalkyl, any of which is substituted or unsubstituted; 
 each R 3  is independently alkylene, alkenylene, or alkynylene, any of which is substituted or unsubstituted; 
 each x is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; 
 each w and y is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20; 
 z is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and 
 each Xaa is independently an amino acid, 
 
         wherein the compound exhibits an intracellular antimicrobial effect against at least one microbe. 
       
     
     
         2 . The compound of  claim 1 , wherein x is 2, 3, or 6. 
     
     
         3 . The compound of  claim 2 , wherein x is 2; R 3  is C 8  alkylene, C 8  alkenylene, or C 8  alkynylene; and the sum of x, w, and y is at least 10. 
     
     
         4 . The compound of  claim 1 , wherein x is 3; R 3  is C 8  alkylene, C 8  alkenylene, or C 8  alkynylene; and the sum of x, w, and y is at least 10. 
     
     
         5 . The compound of  claim 1 , wherein R 3  is substituted with 1, 2, 3, 4, 5, or 6 R 4 , and each R 4  is independently —NH 3  or —OH, wherein each —NH 3  is optionally coupled with another chemical entity. 
     
     
         6 . The compound of  claim 5 , wherein R 3  is substituted with 2 R 4 , and both R 4  are —OH. 
     
     
         7 . The compound of  claim 5 , wherein R 3  is substituted with 2 R 4 , and one R 4  is an optionally substituted —NH 3  and the other R 4  is —OH. 
     
     
         8 . The compound of  claim 1 , wherein at least one Xaa is an amino acid that is a non-natural amino acid. 
     
     
         9 . The compound of  claim 1 , wherein the compound comprises a helix. 
     
     
         10 . The compound of  claim 9 , wherein the helix is an α-helix. 
     
     
         11 . The compound of  claim 1 , wherein R 3  spans from one to two turns on an α-helix. 
     
     
         12 . The compound of  claim 1 , wherein the compound comprises an amino acid sequence that is at least 60% identical to an amino acid sequence selected from the group consisting of: SEQ ID NOs: 1-60. 
     
     
         13 . The compound of  claim 1 , wherein the compound comprises an amino acid sequence that is at least 70% identical to an amino acid sequence selected from the group consisting of: SEQ ID NOs: 1-60. 
     
     
         14 . The compound of  claim 1 , wherein the compound comprises an amino acid sequence that is at least 80% identical to an amino acid sequence selected from the group consisting of: SEQ ID NOs: 1-60. 
     
     
         15 . The compound of  claim 1 , wherein the compound comprises an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of: SEQ ID NOs: 1-60. 
     
     
         16 . The compound of  claim 1 , wherein the compound comprises an amino acid sequence that is any one of: SEQ ID NOs: 1-60. 
     
     
         17 . The compound of  claim 1 , wherein the compound comprises BFStap(i+4)2, BFStap(i+4)3, BFStap(i+4)4, BFStap(i+4)6, BFStap(i+4)7, BFStap(i+4)8, BFStap(i+4)11, BFStap(i+7)1, BFStap(i+7)2, BFStap(i+7)3, BFStap(i+7)4, BFStap(i+7)5, BFStap(i+7)6, BFStap(i+7)7, BFStap(i+7)8, or BFStap(i+7)9. 
     
     
         18 . The compound of  claim 1 , wherein the compound comprises the sequence of BFStap(i+4)7, BFStap(i+4)11, or BFStap(i+7) 6. 
     
     
         19 . The compound of  claim 1 , wherein the antimicrobial effect is a minimum inhibitory concentration between about 0.1 μM and about 50 μM. 
     
     
         20 . The compound of  claim 1 , wherein the antimicrobial effect is a minimum inhibitory concentration between about 0.5 μM and about 20 μM. 
     
     
         21 . The compound of  claim 1 , wherein the microbe is a bacterial organism. 
     
     
         22 . The compound of  claim 21 , wherein the bacterial organism is Gram-positive. 
     
     
         23 . The compound of  claim 21 , wherein the bacterial organism is Gram-negative. 
     
     
         24 . The compound of  claim 21 , wherein the bacterial organism is  Mycobacterium tuberculosis.    
     
     
         25 . A method of treating an infection caused by a microbe, the method comprising administering a therapeutically-effective amount of a compound having the formula: 
       
         
           
           
               
               
           
         
         or a pharmaceutically acceptable salt thereof,
 wherein; 
 each R 1  and R 2  is independently H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkylalkyl, heteroarylalkyl, or heterocyclylalkyl, any of which is substituted or unsubstituted; 
 each R 3  is independently alkylene, alkenylene, or alkynylene, any of which is substituted or unsubstituted; 
 each x is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; 
 each w and y is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20; 
 z is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and 
 each Xaa is independently an amino acid, 
 
         to a subject having, or at risk of having, the infection caused by the microbe, wherein the compound exhibits an intracellular antimicrobial effect against at least one microbe. 
       
     
     
         26 . The method of  claim 25 , wherein the subject is an animal. 
     
     
         27 . The method of  claim 26 , wherein the animal is a mammal. 
     
     
         28 . The method of  claim 26 , wherein the animal is a human. 
     
     
         29 . The method of  claim 25 , wherein the subject is a plant. 
     
     
         30 . The method of  claim 25 , wherein the microbe is a bacterial organism. 
     
     
         31 . The method of  claim 30 , wherein the bacterial organism is Gram-positive. 
     
     
         32 . The method of  claim 30 , wherein the bacterial organism is Gram-negative. 
     
     
         33 . The method of  claim 25 , further comprising administering to the subject a therapeutically-effective amount of an antibiotic. 
     
     
         34 . The method of  claim 33 , wherein the compound and the antibiotic act synergistically to treat the infection. 
     
     
         35 . The method of  claim 33 , wherein the compound and the antibiotic act synergistically to overcome a resistance of the infection to the antibiotic. 
     
     
         36 . A pharmaceutical composition comprising a compound having the formula: 
       
         
           
           
               
               
           
         
         or a pharmaceutically acceptable salt thereof,
 wherein; 
 each R 1  and R 2  is independently H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkylalkyl, heteroarylalkyl, or heterocyclylalkyl, any of which is substituted or unsubstituted; 
 each R 3  is independently alkylene, alkenylene, or alkynylene, any of which is substituted or unsubstituted; 
 each x is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; 
 each w and y is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20; 
 z is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and 
 each Xaa is independently an amino acid, 
 
         wherein the compound exhibits an intracellular antimicrobial effect against at least one microbe.