Nemo coiled coil mimics and methods of using same

A macrostructure comprising a parallel coiled-coil, wherein the parallel coiled-coil comprises:

• a first coil of Formula I and a second coil of Formula II:
• (Formula I)
• (SEQ ID NO:
• 1)
• T1-f0-g0-a1-b1-c1-d1-e1-f1-g1-a2-b2-c2-d2-e2-f2-g2-a3-
• b3-c3-d3-e3-T2
• (Formula II)
• (SEQ ID NO:
• 2)
• T3-g′0-a′1-b′1-c′1-d′1-e′1-f′1-g′1-a′2-b′2-c′2-d′2-
• e′2-f′2-g′2-a′3-b′3-c′3-d′3-e′3-f′3-T4,
• wherein:
• each a1-3, b1-3, c1-3, d1-3, e1-3, f0-2, g0-2, a′1-3, b′1-3, c′1-3, d′1-3, e′1-3, f1-3, and g′0-2 is independently absent or a residue selected from the group consisting of modified or unmodified amino acid residues and analogues thereof;
• one or more of the following residue pairs are covalently bound by a linker:
• g0-e′1, g1-e′2, g2-e′3, d1-d′1, d2-d′2, d3-d′3, a1-a′1, a2-a′2, a3-a′3, e1-g′0, e2-g′1, and e3-g′2;
• each T1 and T3 is independently a point of attachment from a terminal nitrogen to one or more moieties, wherein each moiety is independently H, —PG1, —C(O)R, —C(O)NR2, —C(O)NH2, —R, —C(O)OR, an amino acid or analogue thereof, a peptide, a targeting moiety, or a tag, where PG1 is an amine protecting group and each R is independently hydrogen, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, an aryl, a heteroaryl, a heterocyclyl, an arylalkyl, a peptide, a targeting moiety, or a tag;
• and
• each T2 and T4 is independently a point of attachment from a terminal carbonyl to H, -OPG2, -NPG2, —OR, —OH, —NR2,—NH2, —N(R)C(O)C1-6 alkyl, -N(H)C(O)C1-6 alkyl, an amino acid or analogue thereof, a peptide, a targeting moiety, or a tag, where PG2 is a carboxylic acid protecting group and each R is independently hydrogen, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, an aryl, a heteroaryl, a heterocyclyl, an arylalkyl, a peptide, a targeting moiety, or a tag;
• and wherein:
• the first coil comprises at least fourteen contiguous residues, wherein the at least fourteen contiguous residues have the formula X1-X2-X3-X4_X5-X6-X7-X8-X9-X10-X11-X12-X13-X14;
• the second coil comprises at least fourteen contiguous residues, wherein the at least fourteen contiguous residues have the formula X′1—X′2—X′3—X′4-X's-X′6—X′7—X′8—X′9—X′10—X′11—X′12—X′13—X′14;
• and
• wherein each residue is selected from the groups indicated below (superscript letters indicate each residue's location within Formula I and Formula II;
• residues in the a, a′, d, d′, e, e′, g, and g′ positions can optionally be modified to facilitate attachment of a linker or replaced with a linker)
• First Coil
• Second Coil
• Preferred
• Preferred
• Residue
• Group
• Residue(s)
• Residue
• Group
• Residue(s)
• fX1
• any residue
• Val, Glu
• gX2
• modified or
• Trp
• gX′1
• any residue
• Ala, Glu,
• unmodified
• azidolysine,
• Trp and
• Cys
• analogues
• thereof
• aX3
• any
• Cys, HCys, Leu,
• aX′2
• any
• Cys, HCys, Leu,
• hydrophobic
• Ile, allylleucine,
• hydrophobic
• Ile, allylleucine,
• residue
• Val, allylglycine,
• residue
• Val, allylglycine,
• Thr,
• Thr,
• selenocysteine,
• selenocysteine,
• hexafluoroleucine,
• hexafluoroleucine,
• hexafluorovaline
• hexafluorovaline
• (or analogue of
• (or analogue of
• any of the
• any of the
• preceding
• preceding
• residues)
• residues)
• bX4
• any
• His, Arg
• bX′3
• modified or
• His
• positively
• unmodified
• charged
• His and
• residue
• analogues
• thereof
• cX5
• modified or
• Gln
• cX′4
• any
• Gln, Arg
• unmodified
• positively
• Gln and
• charged
• analogues
• residue,
• thereof
• modified or
• unmodified
• Gln and
• analogues
• thereof
• dX6
• any
• Cys, HCys, Leu,
• dX′5
• any
• Cys, HCys, Leu,
• hydrophobic
• Ile, allylleucine,
• hydrophobic
• Ile, allylleucine,
• residue
• Val, allylglycine,
• residue
• Val, allylglycine,
• Thr,
• Thr,
• selenocysteine,
• selenocysteine,
• hexafluoroleucine,
• hexafluoroleucine,
• hexafluorovaline
• hexafluorovaline
• (or analogue of
• (or analogue of
• any of the
• any of the
• preceding
• preceding
• residues)
• residues)
• eX7
• any residue
• Phe, Tyr, Trp,
• eX′6
• modified or
• Phe
• azidolysine, Cys
• unmodified
• Phe and
• analogues
• thereof
• fX8
• any
• Gln, Glu
• fX′7
• any
• Gln, Arg
• negatively
• positively
• charged
• charged
• residue,
• residue,
• modified or
• modified or
• unmodified
• unmodified
• Gln and
• Gln and
• analogues
• analogues
• thereof
• thereof
• gX9
• modified or
• QCy
• gX′8
• azidolysine
• azidolysine, Glu,
• unmodified
• and
• Arg
• QCy and
• analogues
• analogues
• thereof,
• thereof
• modified or
• unmodified
• Glu and
• analogues
• thereof,
• modified or
• unmodified
• Arg and
• analogues
• thereof
• aX10
• any
• Cys, HCys, Leu,
• aX′9
• any
• Cys, HCys, Leu,
• hydrophobic
• Ile, allylleucine,
• hydrophobic
• Ile, allylleucine,
• residue
• Val, allylglycine,
• residue
• Val, allylglycine,
• Thr,
• Thr,
• selenocysteine,
• selenocysteine,
• hexafluoroleucine,
• hexafluoroleucine,
• hexafluorovaline
• hexafluorovaline
• (or analogue of
• (or analogue of
• any of the
• any of the
• preceding
• preceding
• residues)
• residues)
• bX11
• any residue
• Asp, Arg
• bX′10
• modified or
• Asp
• unmodified
• Asp and
• analogues
• thereof
• cX12
• modified or
• Arg
• cX′11
• any
• Asn, Glu
• unmodified
• negatively
• Arg and
• charged
• analogues
• residue,
• thereof
• modified or
• unmodified
• Asn and
• analogues
• thereof
• dX13
• any
• Cys, HCys, Leu,
• dX′12
• any
• Cys, HCys, Leu,
• hydrophobic
• Ile, allylleucine,
• hydrophobic
• Ile, allylleucine,
• residue
• Val, allylglycine,
• residue
• Val, allylglycine,
• Thr,
• Thr,
• selenocysteine,
• selenocysteine,
• hexafluoroleucine,
• hexafluoroleucine,
• hexafluorovaline
• hexafluorovaline
• (or analogue of
• (or analogue of
• any of the
• any of the
• preceding
• preceding
• residues)
• residues)
• eX14
• azidolysine
• azidolysine, Glu,
• eX′13
• modifiedor
• Ile
• and
• Arg
• unmodified
• analogues
• Ile and
• thereof,
• analogues
• modified or
• thereof
• unmodified
• Glu and
• analogues
• thereof,
• modified or
• unmodified
• Arg and
• analogues
• thereof
• fX′14
• modified or
• Arg
• unmodified
• Arg and
• positively
• charged
• analogues
• thereof