Lipid-peptide-polymer conjugates and nanoparticles thereof

Abstract

The present invention provides a conjugate having a peptide with from about 10 to about 100 amino acids, wherein the peptide adopts a helical structure. The conjugate also includes a first polymer covalently linked to the peptide, and a hydrophobic moiety covalently linked to the N-terminus of the peptide, wherein the hydrophobic moiety comprises a second polymer or a lipid moiety. The present invention also provides helix bundles form by self-assembling the conjugates, and particles formed by self-assembling the helix bundles. Methods of preparing the helix bundles and particles are also provided.

1 .- 17 . (canceled) 
     
     
         18 . A conjugate comprising:
 a three-helix bundle-forming peptide having from about 10 to about 100 amino acids;   a first polymer covalently linked to the three-helix bundle-forming peptide, wherein the first polymer is linked to the three-helix bundle-forming peptide at an amino acid other than the N- or C-terminus; and   a hydrophobic moiety covalently linked to the N-terminus of the peptide, wherein the hydrophobic moiety comprises a second polymer or a lipid moiety comprising from 1 to 6 C 10-20  alkyl groups.   
     
     
         19 . The conjugate of  claim 18 , wherein the amino acids in the three-helix bundle-forming peptide are characterized by a heptad periodicity, -abcdefg-,
 wherein the amino acids at positions a and d are hydrophobic amino acids, and   wherein the amino acids at positions e and g form salt bridges between adjacent helices.   
     
     
         20 . The conjugate of  claim 19 , wherein each amino acid at position a and position d is independently selected from the group consisting of valine, leucine, isoleucine, methionine, phenylalanine, and tryptophan. 
     
     
         21 . The conjugate of  claim 19 , wherein each amino acid at position e and position g is independently selected from the group consisting of lysine, arginine, histidine, aspartate, and glutamate. 
     
     
         22 . The conjugate of  claim 18 , wherein the first polymer is a hydrophilic polymer. 
     
     
         23 . The conjugate of  claim 18 , wherein the first polymer is polyethyleneglycol. 
     
     
         24 . The conjugate of  claim 18 , wherein the second polymer comprises polybutadiene. 
     
     
         25 . The conjugate of  claim 18 , wherein the lipid moiety comprises from 1 to 6 C 10-20  alkyl groups. 
     
     
         26 . The conjugate of  claim 18 , wherein the lipid moiety comprises 1, 2, or 4 C 10-20  alkyl groups. 
     
     
         27 . The conjugate of  claim 18 , further comprising an amino acid residue covalently linked to the C-terminus of the three-helix bundle-forming peptide. 
     
     
         28 . The conjugate of  claim 27 , wherein the amino acid residue comprises a member selected from the group consisting of GGG, HHH, KK, EE, RGD and AYSSGAPPMPPF. 
     
     
         29 . The conjugate of  claim 18 , wherein
 the amino acids in the three-helix bundle-forming peptide are characterized by a heptad periodicity, -abcdefg-, wherein   each amino acid at position a and position d is independently selected from the group consisting of valine, leucine, isoleucine, methionine, phenylalanine, and tryptophan, and   each amino acid at position e and position g is independently selected from the group consisting of lysine, arginine, histidine, aspartate, and glutamate;   the first polymer comprises polyethylene glycol;   the hydrophobic moiety comprises the lipid moiety which comprises lysine and two C 16  alkyl chains; and   an amino acid residue of from 2 to about 20 amino acids, covalently linked to the C-terminus of the three-helix bundle-forming peptide.   
     
     
         30 . A helix bundle comprising 3 conjugates of  claim 18 . 
     
     
         31 . A particle comprising from about 20 to about 200 conjugates of  claim 18 . 
     
     
         32 . The particle of  claim 31 , further comprising at least one member selected from the group consisting of a therapeutic agent, a diagnostic agent, DNA, and an oligonucleotide. 
     
     
         33 . A method of forming a particle of  claim 31 , the method comprising:
 contacting a plurality of conjugates of  claim 18  such that the conjugates self-assemble to form the particles of  claim 31 .   
     
     
         34 . The method of  claim 33 , wherein the conjugates are at a concentration of from about 1 nM to about 1 M.