Multivalent ligands which modulate angiogenesis

Claim 2

A peptide having the amino acid sequence set forth in SEQ ID NO.: 5, SEQ ID NO.: 6, a subsequence of SEQ ID NO.: 5 having at least ten amino acid residues or a subseqence of SEQ ID NO.: 6 having at least ten amino acid residues.

Claim 3

The peptide of claim 1 wherein the N-terminus is substituted with an amine protecting group and/or the C-terminus is substituted with a carboxylic acid protecting group.

Claim 4

The peptide of claim 1 wherein the peptide has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 5 or SEQ ID NO.: 6 having from ten to fifteen amino acid residues.

Claim 5

The peptide of claim 1 wherein the peptide has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 5 or SEQ ID NO.: 6 having from about ten to about fifteen amino acid residues, wherein one, two or three amino acid residues in the sequence of the peptide can differ from the amino acid residue(s) in the corresponding position(s) of the subsequence.

Claim 6

The peptide of claim 1 wherein the peptide has an amino acid sequence set forth in SEQ ID NO.: 5, SEQ ID NO.: 6, a subsequence of SEQ ID NO.: 5 having at least sixteen amino acid residues or a subsequence of SEQ ID NO.: 6 having at least sixteen amino acid residues.

Claim 7

The peptide of claim I wherein the peptide has an amino acid sequence set forth in SEQ ID NO.: 5, SEQ ID NO. 6, a subsequence of SEQ ID NO.: 5 having at least sixteen amino acid residues or a subsequence of SEQ ID NO.: 6 having at least sixteen amino acid residues, wherein one, two, three or four amino acid residues in the sequence of the peptide can differ from the amino acid residue(s) in the corresponding positions of SEQ ID NO.: 5, SEQ ID NO. 6, the subsequence of SEQ ID NO.: 5 or the subsequence of SEQ ID NO.: 6.

Claim 8

The peptide of claim 1 wherein the peptide has an amino acid sequence as set forth in SEQ ID NOS.: 3 or 4.

Claim 9

A multivalent ligand set forth in the following structural formula: wherein: B is a multilinker backbone; n is an integer from two to about twenty; each L is a covalent bond or linking group; each P is a peptide, wherein at least two of the peptides are a peptide derivative of an AHR of an angiogenic protein, a hybrid peptide, a peptide derivative of a hybrid peptide, or a combination thereof, wherein: a) said multivalent ligand has angiogenic activity and b) each peptide and each linker or covalent bond is independently chosen.

Claim 10

A multivalent ligand set forth in the following structural formula: wherein: B is a multilinker backbone; n is an integer from two to about twenty; each L is a covalent bond or linking group; each P is a peptide, wherein at least two of the peptides are a peptide derivative of an AHR of an angiogenic protein, a hybrid peptide, a peptide derivative of a hybrid peptide, or a combination thereof, wherein: a) said multivalent ligand has angiogenic activity and b) each peptide and each linker or covalent bond is independently chosen.

Claim 11

The multivalent ligand of claim 8 wherein each L is the same and each peptide is the same.

Claim 12

The multivalent ligand of claim 9 wherein the multilinker backbone is a polymer with two or more reactive groups.

Claim 13

The multivalent ligand of claim 9 wherein the multilinker backbone is a polypeptide.

Claim 14

The multivalent ligand of claim 11 wherein the multilinker backbone is a cascading or pennant polypeptide.

Claim 15

The mutlivalent ligand of claim 12 wherein each linker is a covalent bond, a glycine residue or a residue of glycine-glycine.

Claim 16

The multivalent ligand of claim 13 wherein the cascading or pennant polypeptide is a polylysine.

Claim 17

The multivalent ligand of claim 13 wherein the cascading or pennant polypeptide is a polyaspartic acid or a polyglutamic acid.

Claim 18

The multivalent ligand of claim 11 wherein the backbone contains a cysteine residue, wherein a bond is formed between the thiol group in the cysteine side chain and a linker or peptide.

Claim 19

The multivalent ligand of claim 14 wherein each peptide is a peptide derivative of the angiogenic homology region of an angiogenic protein.

Claim 20

The multivalent ligand of claim 17 wherein the peptide derivative has between about ten and about twenty-eight amino acid residues.

Claim 21

The multivalent ligand of claim 18 wherein the AHR is set forth in SEQ ID NO.: 1 or SEQ ID NO.: 2; and wherein the peptide derivative has between about ten and about twenty-eight amino acid residues or amino acid residue analogs.

Claim 22

The multivalent ligand of claim 19 wherein: a) each peptide derivative has between ten and fifteen amino acid residues; and b) each peptide derivative has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 1 or a subsequence of SEQ ID NO.: 2.

Claim 23

The multivalent ligand of claim 19 wherein: a) each peptide derivative has between ten and fifteen amino acid residues; and b) each peptide derivative has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 1 or a subsequence of SEQ ID NO.: 2, with the proviso that one, two or three amino acid residues in the sequence of c) the peptide derivative can differ from the amino acid residue(s) in the corresponding position(s) of the subsequence.

Claim 24

The multivalent ligand of claim 19 wherein: a) each peptide derivative has between fifteen and twenty-eight amino acid residues; and b) each peptide derivative has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 1 or a subsequence of SEQ ID NO.: 2.

Claim 25

The multivalent ligand of claim 19 wherein: a) each peptide derivative has between fifteen and twenty-eight amino acid residues; and b) each peptide derivative has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 1 or a subsequence of SEQ ID NO.: 2, with the proviso that one, two three or four amino acid residues in the sequence of the peptide derivative can differ from the amino acid residue(s) in the corresponding position(s) of the subsequence.

Claim 26

The multivalent ligand of claim 19 wherein: a) the multivalent ligand has four peptide derivatives set forth in SEQ ID NO.: 1 or SEQ ID NO.: 2; and b) the multilinker backbone is a pennant polylysine consisting of three lysine residues.

Claim 27

The multivalent ligand of claim 14 wherein each peptide comprises a sequence of amino acids AA 1 through AA 28 or a subsequence thereof having at least ten amino acids, wherein: AA 1 is lysine, ornithine, arginine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline, 2-amino-4-guanidinobutanoic acid, glycine or alanine; AA 2 is aspartic acid, asparagine, glutamic acid, glutamine, leucine, valine, isoleucine, methionine or a substituted or unsubstituted aliphatic or aryl ester of glutamic acid or aspartic acid; AA 3 is phenylalanine, alanine, tyrosine, tryptophan or glycine; AA 4 is threonine, glycine, alanine, cysteine or serine; AA 5 is alanine, threonine, glycine, cysteine or serine; AA 6 is phenylalanine, tyrosine or tryptophan; AA 7 is arginine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline or 2-amino-4-guanidinobutanoic acid; AA 8 is tryptophan, alanine, phenylalanine, tyrosine or glycine; AA 9 is arginine, phenylalanine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline, 2-amino-4-guanidinobutanoic acid, lysine, ornithine, tyrosine or tryptophan; AA 10 is leucine, isoleucine, methionine or valine; AA 11 is serine, threonine or alanine; AA 12 is histidine, serine, threonine, cysteine, lysine or ornithine; AA 13 is arginine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline or 2-amino-4-guanidinobutanoic acid; AA 14 is proline, leucine, valine, isoleucine or methionine; AA 15 is lysine, glutamine, histidine, ornithine, asparagine, arginine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline or 2-amino-4-guanidinobutanoic acid; AA 16 is threonine, aspartic acid, serine, glutamic acid or a substituted or unsubstituted aliphatic or aryl ester of glutamic acid or aspartic acid; AA 17 is glycine, leucine, alanine, valine, isoleucine or methionine; AA 18 is phenylalanine, tyrosine or tryptophan; AA 19 is isoleucine, serine, valine, leucine, methionine, cysteine or threonine; AA 20 is arginine, isoleucine, ornithine, lysine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline, 2-amino-4-guanidinobutanoic acid, leucine, valine or methionine; AA 21 is methionine, isoleucine, leucine or valine; AA 22 is valine, arginine, leucine, isoleucine, methionine, ornithine, lysine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline or 2-amino-4-guanidinobutanoic acid; AA 23 is methionine, arginine, leucine, isoleucine, valine, ornithine, lysine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline or 2-amino-4-guanidinobutanoic acid; AA 24 is phenylalanine, alanine, tyrosine, tryptophan or glycine; AA 25 is aspartic acid, asparagine, glutamic acid, glutamine or a substituted or unsubstituted aliphatic or aryl ester of glutamic acid or aspartic acid; AA 26 is glycine, arginine, alanine, ornithine, lysine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline or 2-amino-4-guanidinobutanoic acid; AA 27 is lysine, alanine, arginine, glycine, serine, ornithine, arginine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline or 2-amino-4-guanidinobutanoic acid; and AA 28 is lysine, alanine, arginine, glycine, serine, ornithine, arginine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline or 2-amino-4-guanidinobutanoic acid; wherein each peptide and each linker or covalent bond is independently chosen and the multivalent ligand has angiogenic activity.

Claim 28

The multivalent ligand of claim 25 wherein the peptide has a equence comprising a subsequence of SEQ ID NO.: 1 and a subsequence of SEQ ID NO.: 2, wherein each subsequence has from five to fifteen amino acid residues.

Claim 29

The multivalent linker of claim 26 wherein the peptide has a sequence consisting of a subsequence of SEQ ID NO: 1 and a subsequence of SEQ ID NO.: 2, wherein each subsequence has from five to fifteen amino acid residues.

Claim 30

The multivalent linker of claim 27 wherein the subsequence of SEQ ID NO.: 1 and the subsequence of SEQ ID NO.: 2 each have about nine amino acid residues.

Claim 31

The multivalent linker of claim 25 wherein: a) the multilinker backbone is a pennant polylysine consisting of three lysine residues; b) the multivalent ligand comprises four peptides set forth in SEQ ID NO. 3, SEQ ID NO.: 4 or SEQ ID NOS. 9-12; and c) each linker is a covalent bond.

Claim 32

The multivalent ligand of claim 14 wherein each peptide is a hybrid peptide or a peptide derivative thereof.

Claim 33

The multivalent ligand of claim 30 wherein the peptide is a set forth in SEQ ID NO.: 3, SEQ ID NO.: 4 or SEQ ID NOS. 9-12 or a peptide derivative of SEQ ID NO.: 3, SEQ ID NO.: 4 or SEQ ID NOS. 9-12.

Claim 34

The multivalent ligand of claim 31 wherein the peptide derivative has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 3, SEQ ID NO.: 4 or SEQ ID NOS. 9-12 having from ten to fifteen amino acid residues.

Claim 35

The multivalent ligand of claim 31 wherein the peptide derivative has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 3, SEQ ID NO.: 4 or SEQ ID NOS. 9-12 having from about ten to about fifteen amino acid residues, wherein one, two or three amino acid residue(s) in the sequence of the peptide derivative can differ from the amino acid residue(s) in the corresponding position(s) of the subsequence.

Claim 36

The multivalent ligand of claim 31 wherein the peptide derivative has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 3, SEQ ID NO.: 4 or SEQ ID NOS. 9-12, said subsequence having from sixteen to twenty-eight amino acid residues.

Claim 37

The multivalent ligand of claim 31 wherein the peptide derivative has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 3, SEQ ID NO.: 4, or SEQ ID NOS. 9-12 said subsequence having from about sixteen to about twenty-eight amino acid residues, wherein one, two, three or four amino acid residue(s) in the sequence of the peptide derivative can differ from the amino acid residue(s) in the corresponding position(s) of the subsequence.

Claim 40

The method of claim 36 wherein the multivalent ligand is anti-angiogenic and the subject is being treated for cancer.

Claim 41

The method of claim 36 wherein the multivalent ligand is pro-angiogenic.

Claim 42

The method of claim 36 wherein: a) the multivalent peptide backbone is a pennant or cascading polylysine; b) each linker and each polypeptide is the same; c) and each linker is a covalent bond, a glycine residue or a glycine-glycine residue.

Claim 43

The method of claim 39 wherein each peptide is a peptide derivative of the AHR of an angiogenic protein.

Claim 44

The method of claim 40 wherein: a) the angiogenic homology region is set forth in SEQ ID NO.: 1 or SEQ ID NO.: 2; and b) the peptide derivative has between about ten and about twenty-eight amino acid residues or amino acid residue analogs.

Claim 45

The method of claim 41 wherein: a) each peptide derivative has between ten and fifteen amino acid residues; and b) each peptide derivative has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 1 or a subsequence of SEQ ID NO.: 2.

Claim 46

The method of claim 41 wherein: a) each peptide derivative has between ten and fifteen amino acid residues; and b) each peptide derivative has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 1 or a subsequence of SEQ ID NO.: 2, with the proviso that one, two or three amino acid residues in the sequence of c) the peptide derivative can differ from the amino acid residue(s) in the corresponding position(s) in the subsequence.

Claim 47

The method of claim 41 wherein: a) each peptide derivative has between fifteen and twenty-eight amino acid residues; and b) each peptide derivative has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 1 or a subsequence of SEQ ID NO.: 2.

Claim 48

The method of claim 41 wherein: a) each peptide derivative has between fifteen and twenty-eight amino acid residues; and b) each peptide derivative has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 1 or a subsequence of SEQ ID NO.: 2, with the proviso that one, two, three or four amino acid residues in the sequence of the peptide derivative can differ from the amino acid residue(s) in the corresponding position of the subsequence.

Claim 49

The method of claim 41 wherein the multilinker backbone is a pennant polylysine consisting of four amino acids, and the peptide derivative is set forth in SEQ ID NO.: I or SEQ ID NO.: 2.

Claim 50

The method of claim 39 wherein the peptide comprises a sequence of amino acids AA 1 through AA 28 or a subsequence thereof having at least ten amino acids, wherein: AA 1 is lysine, ornithine, arginine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline, 2-amino-4-guanidinobutanoic acid, glycine or alanine; AA 2 is aspartic acid, asparagine, glutamic acid, glutamine, leucine, valine, isoleucine, methionine or a substituted or unsubstituted aliphatic or aryl ester of glutamic acid or aspartic acid; AA 3 is phenylalanine, alanine, tyrosine, tryptophan, leucine, isoleucine, methionine, valine or glycine; AA 4 is threonine, glycine, alanine, cysteine or serine; AA 5 is alanine, threonine, glycine, cysteine or serine; AA 6 is phenylalanine, tyrosine or tryptophan; AA 7 is arginine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline or 2-amino-4-guanidinobutanoic acid; AA 8 tryptophan, alanine, phenylalanine, tyrosine or glycine; AA 9 is arginine, phenylalanine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline, 2-amino-4-guanidinobutanoic acid, lysine, ornithine, tyrosine or tryptophan; AA 10 is leucine, isoleucine, methionine or valine; AA 11 is serine, threonine or alanine; AA 12 is histidine, serine, threonine, cysteine, lysine or ornithine; AA 13 is arginine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline or 2-amino-4-guanidinobutanoic acid; AA 14 is proline, leucine, valine, isoleucine or methionine; AA 15 is lysine, glutamine, histidine, ornithine, asparagine, arginine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline or 2-amino-4-guanidinobutanoic acid; AA 16 is threonine, aspartic acid, serine, glutamic acid or a substituted or unsubstituted aliphatic or aryl ester of glutamic acid or aspartic acid; AA 17 is glycine, leucine, alanine, valine, isoleucine or methionine; AA 18 is phenylalanine, tyrosine or tryptophan; AA 19 is isoleucine, serine, valine, leucine, methionine, cysteine or threonine; AA 20 is arginine, isoleucine, ornithine, lysine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline, 2-amino-4-guanidinobutanoic acid, leucine, valine or methionine; AA 21 is methionine, isoleucine, leucine or valine; AA 22 is valine, arginine, leucine, isoleucine, methionine, ornithine, lysine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline or 2-amino-4-guanidinobutanoic acid; AA 23 is methionine, arginine, leucine, soleucine, valine, ornithine, lysine, N-nitroarginine, β-cycloarginine, y-hydroxyarginine, N-amidinocitruline or 2-amino-4-guanidinobutanoic acid; AA 24 is phenylalanine, alanine, tyrosine, tryptophan or glycine; AA 25 is aspartic acid, asparagine, glutamic acid, glutamine or a substituted or unsubstituted aliphatic or aryl ester of glutamic acid or aspartic acid; AA 26 is glycine, arginine, alanine, ornithine, lysine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline or 2-amino-4-guanidinobutanoic acid; AA 27 is lysine, alanine, arginine, glycine, serine, ornithine, arginine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline or 2-amino-4-guanidinobutanoic acid; and AA 28 is lysine, alanine, arginine, glycine, serine, ornithine, arginine, N-nitroarginine, β-cycloarginine, γ-hydroxyarginine, N-amidinocitruline or 2-amino-4-guanidinobutanoic acid; wherein each peptide and each linker or covalent bond is independently chosen and the multivalent ligand has angiogenic activity.

Claim 51

The method of claim 47 wherein the peptide has a sequence comprising a subsequence of SEQ ID NO.: 1 and a subsequence of SEQ ID NO.: 2, wherein each subsequence has from five to fifteen amino acid residues.

Claim 52

The method of claim 48 wherein the peptide has a sequence consisting of a subsequence of SEQ ID NO.: 1 and a subsequence of SEQ ID NO.: 2, wherein each subsequence has from five to fifteen amino acid residues.

Claim 53

The method of claim 49 the subsequence of SEQ ID NO.: 1 and the subsequence of SEQ ID NO.: 2 each have at least nine amino acid residues.

Claim 54

The method of claim 49 wherein the peptide is a hybrid peptide or a peptide derivative thereof.

Claim 55

The method of claim 51 wherein the hybrid peptide is set forth in SEQ ID NO.: 3, SEQ ID NO.: 4 or SEQ ID NOS. 9-12 or is a peptide derivative thereof.

Claim 56

The method of claim 52 wherein the peptide derivative of SEQ ID NO.: 3, SEQ ID NO.: 4 or SEQ ID NOS. 9-12 have between about ten and about twenty-eight amino acid residues or amino acid residue analogs.

Claim 57

The method of claim 53 wherein the peptide derivative has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 3, SEQ ID NO.: 4 or SEQ ID NOS. 9-12 having from ten to fifteen amino acid residues.

Claim 58

The method of claim 53 wherein the peptide derivative has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 3, SEQ ID NO.: 4 or SEQ ID NOS. 9-12 having from about ten to about fifteen amino acid residues, wherein one, two or three amino acid residues in the sequence of the peptide derivative can differ from the amino acid residue(s) at the corresponding positions(s) of the subsequence.

Claim 59

The method of claim 53 wherein the peptide derivative has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 3 or a subsequence of SEQ ID NO.: 4 or SEQ ID NOS. 9-12, said subsequence having from sixteen to twenty-eight amino acid residues.

Claim 60

The method of claim 53 wherein the peptide derivative has an amino acid sequence set forth in a subsequence of SEQ ID NO.: 3, a subsequence of SEQ ID NO.: 4 or SEQ ID NOS. 9-12, said subsequence having from about sixteen to about twenty-eight amino acid residues, wherein one, two, three or four amino acid residues in the sequence of the peptide derivative can differ from the amino acid residue(s) at the corresponding positions(s) of the subsequence.

Claim 61

The method of claim 53 wherein the peptide derivative is set forth in SEQ ID NO.: 3, SEQ ID NO.: 4 or SEQ ID NOS. 9-12 and wherein the multilinker backbone is a pennant polylysine consisting of three lysine residues.

Claim 62

A polypeptide multivalent ligand set forth in the following structural formula: wherein: m is an integer from zero to about twenty; each S is a peptide spacer having from about five to about thirty amino acids; each P is a peptide having from about ten to about forty amino acid residues, wherein at least two of the peptides are peptide derivatives of an AHR of an angiogenic protein, a hybrid peptide or a peptide derivative of a hybrid peptide, wherein: a) said polypeptide multivalent ligand has angiogenic activity; and b) each peptide and each peptide spacer is independently chosen.

Claim 63

A polypeptide multivalent ligand set forth in the following structural formula: wherein: m is an integer from zero to about twenty; each S is a peptide spacer having from about five to about thirty amino acids; each P is a peptide having from about ten to about forty amino acid residues, wherein at least two of the peptides are peptide derivatives of an AHR of an angiogenic protein, a hybrid peptide or a peptide derivative of a hybrid peptide, wherein: a) said polypeptide multivalent ligand has angiogenic activity; and b) each peptide and each peptide spacer is independently chosen.