Cell-penetrating compstatin analogs and uses thereof

Abstract

In some aspects, the present invention cell-penetrating compstatin analog and compositions comprising cell-penetrating compstatin analog. In some aspects, the invention further provides methods of using cell-penetrating compstatin analogs treat a complement-mediated disorder. e.g., to inhibit complement-mediated damage to a cell, tissue, or organ, to inhibit production or release of biologically active C3 cleavage products.

1 . A compound comprising a compstatin analog moiety and a cell penetrating moiety (CPM). 
     
     
         2 . The compound of  claim 1 , wherein the CPM comprises a cell penetrating peptide. 
     
     
         3 . The compound of  claim 1 , wherein the CPM comprises a cationic cell penetrating peptide. 
     
     
         4 . The compound of  claim 1 , wherein the compstatin analog moiety is linked to the CPM via a linker. 
     
     
         5 - 6 . (canceled) 
     
     
         7 . The compound of  claim 1 , wherein the CPM comprises or consists of a cell penetrating peptide the sequence of which comprises or consists of or is derived from a sequence set forth in Table 2. 
     
     
         8 . The compound of  claim 1 , wherein wherein the compstatin analog moiety comprises a cyclic peptide having a core sequence of X′aa-Gln-Asp-Xaa-Gly (SEQ ID NO: 3), where X′aa and Xaa are selected from Trp and analogs of Trp. 
     
     
         9 . The compound of  claim 1 , wherein the compstatin analog moiety comprises a cyclic peptide having a core sequence of X′aa-Gln -Asp-Xaa-Gly-X″aa (SEQ ID NO: 4), where X′aa and Xaa are each independently selected from Trp and analogs of Trp, and X″aa is selected from His, Ala, single methyl unbranched amino acids, Phe, Trp, and analogs of Trp. 
     
     
         10 . (canceled) 
     
     
         11 . The compound of  claim 1 , wherein the compstatin analog is a compound that comprises a cyclic peptide having a sequence of X′aa1-X′aa2-X′aa3-X′aa4-Gln-Asp-Xaa-Gly-X″aa1-X″aa2-X″aa3-X″aa4-X″aa5 (SEQ ID NO: 5), where X′aa4 and Xaa are selected from Trp and analogs of Trp, wherein X′aa1, X′aa2, X′aa3, X″aa1, X″aa2, X″aa3, X″aa4, and X″aa5, are independently selected from among amino acids and amino acid analogs, wherein the peptide is cyclized via a bond between X′aa2 and X″aa4. 
     
     
         12 - 13 . (canceled) 
     
     
         14 . The compound of  claim 1 , wherein the compstatin analog is a compound that comprises a cyclic peptide having a sequence: 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 6) 
                 
                     
                   Xaa1 - Cys -Val - Xaa2 - Gln - Asp - Xaa2* - 
                 
                     
                     
                 
                     
                   Gly - Xaa3 - His - Arg - Cys - Xaa4; 
                 
             
                
                
                
                
               
            
           
         
       
       wherein:
 Xaa1 is Ile, Val, Leu, B 1 —Ile, B 1 —Val, B 1 -Leu or a dipeptide comprising Gly-Ile or B 1 -Gly-Ile, and B 1  represents a first blocking moiety; 
 Xaa2 and Xaa2* are independently selected from Trp and analogs of Trp; 
 Xaa3 is His, Ala or an analog of Ala, Phe, Trp, or an analog of Trp; 
 Xaa4 is L-Thr, D-Thr, Ile, Val, Gly, a dipeptide selected from Thr-Ala and Thr-Asn, or a tripeptide comprising Thr-Ala-Asn, wherein a carboxy terminal —OH of any of the L-Thr, D-Thr, Ile, Val, Gly, Ala, or Asn optionally is replaced by a second blocking moiety B 2 ; and 
 the two Cys residues are joined by a disulfide bond. 
 
     
     
         15 . The compound of  claim 14 , wherein
 Xaa1 is Ile, Val, Leu, Ac-Ile, Ac-Val, Ac-Leu or a dipeptide comprising Gly-Ile or Ac-Gly-Ile;
 Xaa2 and Xaa2* are independently selected from Trp and analogs of Trp; 
 Xaa3 is His, Ala or an analog of Ala, Phe, Trp, or an analog of Trp; 
 Xaa4 is L-Thr, D-Thr, Ile, Val, Gly, a dipeptide selected from Thr-Ala and Thr-Asn, or a tripeptide comprising Thr-Ala-Asn, wherein a carboxy terminal —OH of any of the L-Thr, D-Thr, Ile, Val, Gly, Ala, or Asn optionally is replaced by —NH2. 
   
     
     
         16 . (canceled) 
     
     
         17 . The compound of  claim 14 , wherein Xaa2 is an analog of Trp comprising a substituted or unsubstituted bicyclic aromatic ring component or two or more substituted or unsubstituted monocyclic aromatic ring components. 
     
     
         18 . (canceled) 
     
     
         19 . The compound of  claim 14 , wherein Xaa2 is an analog of Trp comprising a lower alkoxy or lower alkyl substituent at the 1 or 5 position of tryptophan or a halogen substituent at the 5 or 6 position of tryptophan. 
     
     
         20 . The compound of  claim 19 , wherein Xaa2 is an analog of Trp comprising a lower alkoxy or lower alkyl substituent at the 1 or 5 position of tryptophan or a halogen substituent at the 5 or 6 position of tryptophan and Xaa2* is Trp. 
     
     
         21 . The compound of  claim 1 , wherein the compstatin analog comprises a cyclic peptide having a sequence selected from the group consisting of: SEQ ID NOs: 9-36. 
     
     
         22 . The compound of  claim 1 , wherein the compstatin analog comprises a cyclic peptide having a sequence selected from the group consisting of: SEQ ID NOs: 14, 21, 28, 29, 32, 33, 34, or 36. 
     
     
         23 - 32 . (canceled) 
     
     
         33 . A method of inhibiting activation of primate complement component 3 (C3) comprising contacting said C3 with a compound of  claim 1 . 
     
     
         34 - 35 . (canceled) 
     
     
         36 . A method of reducing the sensitivity of a primate cell or subject to a complement activation stimulus, the method comprising contacting the primate cell or subject with the compstatin analog of  claim 1 . 
     
     
         37 . The method of  claim 36 , wherein the cell or subject is human. 
     
     
         38 . The method of  claim 37 , wherein the method comprises administering the compstatin analog to a subject. 
     
     
         39 . A method of treating a subject in need of treatment for a complement-mediated disorder, the method comprising administering the compstatin analog of  claim 1  to the subject. 
     
     
         40 - 43 . (canceled) 
     
     
         44 . A composition comprising (a) an isolated cell, tissue, or organ; and (b) a cell penetrating compstatin analog. 
     
     
         45 . The composition of  claim 44 , wherein the isolated cell is a liver cell, endothelial cell, epithelial cell, or immune system cell. 
     
     
         46 . A method of characterizing a cell-penetrating compstatin analog (CPCA), the method comprising: (a) contacting a cell that expresses primate C3 with a CPCA in vitro; and (b) assessing the activity of the CPCA. 
     
     
         47 . The method of  claim 46 , wherein assessing the activity of the CPCA comprises exposing the cell to a stimulus that would, in the absence of a CPCA, cause release of C3 cleavage products from the cell; and measuring the amount of one or more C3 cleavage products released from the cell.