Myosin light chain kinase inhibitors and methods of use

Abstract

Disclosed are inhibitors of myosin light chain kinase, pharmaceutical compositions and kits comprising the inhibitors and methods of use.

1 . A method of inhibiting myosin light chain kinase in a cell comprising myosin light chain kinase comprising contacting the cell with a myosin light chain kinase inhibitor in an amount effective to inhibit myosin light chain kinase activity in the cell, the inhibitor having the general formula:
   A-B-C   
       wherein B is covalently bonded to A and C and wherein:
 (a) A and C each comprise at least two basic amino acids; and 
 (b) B comprises Xaa1-Xaa2-Xaa3 wherein 
 (i) Xaa1 is selected from the group consisting of Tyr, Val, Lys, Gln, Phe, Ser, Pro, Thr, Asn, and Arg; 
 (ii) Xaa2 is covalently bonded to Xaa1, and is selected from the group consisting of Lys, Val, Thr, Trp, His, Met, Asn, Ala, Glu, Phe, Gln, and Arg; and 
 (iii) Xaa3 is covalently bonded to Xaa2, and is selected from the group consisting of Ala, Asp, Glu, Phe, Gly, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, and Tyr; and wherein 
 (c) at least one amino acid is a D-amino acid, or the inhibitor comprises at least one non-hydrolyzable bond. 
 
     
     
         2 . The method of  claim 1 , wherein the cell is selected from the group consisting of an epithelial cell, an endothelial cell, and a smooth muscle cell. 
     
     
         3 . The method of  claim 1 , wherein the cell is a mammalian cell. 
     
     
         4 . The method of  claim 3 , wherein the mammalian cell is contacted in vivo. 
     
     
         5 . The method of  claim 1 , wherein the cell is contacted with the inhibitor by contacting the cell with a pharmaceutical composition comprising the inhibitor and a pharmaceutically acceptable carrier. 
     
     
         6 . A method of altering permeability of an epithelial tight junction in vivo comprising administering to a mammal in need thereof an amount of a myosin light chain kinase inhibitor effective to alter permeability, the inhibitor having the general formula:
   A-B-C   
       wherein B is covalently bonded to A and C and wherein:
 (a) A and C each comprise at least two basic amino acids; and 
 (b) B comprises Xaa1-Xaa2-Xaa3 wherein 
 (i) Xaa1 is selected from the group consisting of Tyr, Val, Lys, Gln, Phe, Ser, Pro, Thr, Asn, and Arg; 
 (ii) Xaa2 is covalently bonded to Xaa1, and is selected from the group consisting of Lys, Val, Thr, Trp, His, Met, Asn, Ala, Glu, Phe, Gln, and Arg; and 
 (iii) Xaa3 is covalently bonded to Xaa2, and is selected from the group consisting of Ala, Asp, Glu, Phe, Gly, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, and Tyr; and wherein 
 (c) at least one amino acid is a D-amino acid, or the inhibitor comprises at least one non-hydrolyzable bond.