Vascular endothelial growth factor receptor targeting peptide-elastin fusion polypeptides and their self-assembled nanostructures

Patent metadata

Field	Value
Publication number	US-2018208642-A1
Application number	US-201615744278-A
Country	US
Kind code	A1
Filing date	Oct 19, 2016
Priority date	Apr 7, 2016
Publication date	Jul 26, 2018
Grant date	—

Abstract

Disclosed is a fusion polypeptide for inhibiting neovascularization, including a peptide specifically binding to vascular endothelial growth factor (VEGF) receptors, and a hydrophilic elastin-based polypeptide (hydrophilic EBP) linked to the peptide.

First claim

1 . A fusion polypeptide for inhibiting neovascularization, comprising:
a peptide specifically binding to vascular endothelial growth factor (VEGF) receptors; and a hydrophilic elastin-based polypeptide (hydrophilic EBP) linked to the peptide.

2 . The fusion polypeptide according to claim 1 , wherein the peptide specifically binds to VEGF receptor Flt1 or Flk-1/KDR.

3 . The fusion polypeptide according to claim 2 , wherein the peptide is an anti-Flt1 peptide [SEQ ID NO. 38].

4 . The fusion polypeptide according to claim 1 , wherein the hydrophilic EBP is composed of an amino acid sequence represented by Formula 1 or 2 below:

Formula 1

[SEQ ID NO. 1]n;

or

Formula 2

[SEQ ID NO. 2]n, wherein

SEQ ID NO. 1 is consisted of

[VPGXG VPGXG VPGXG VPGXG VPGXG VPGXG];

SEQ ID NO. 2 is consisted of

[VPAXG VPAXG VPAXG VPAXG VPAXG VPAXG];

n is an integer of 1 or more, and represents the number of repeats of SEQ ID NO. 1 or SEQ ID NO. 2; and
X is an amino acid other than proline, is selected from any natural or artificial amino acids when the pentapeptide VPGXG or VPAXG is repeated, and at least one of X is a hydrophilic amino acid.

5 . The fusion polypeptide according to claim 4 , wherein the hydrophilic EBP is consisted of an amino acid sequence represented by Formula 1 or 2:
in Formula 1, n is 1, and each X of the pentapeptide repeats is consisted of, A (Ala), G (Gly), and I (Ile) in a ratio of 1:4:1 [SEQ ID NO. 20]; K (Lys), G (Gly), and I (Ile) in a ratio of 1:4:1 [SEQ ID NO. 22]; D (Asp), G (Gly), and I (Ile) in a ratio of 1:4:1 [SEQ ID NO. 24]; or E (Glu), G (Gly), and I (Ile) in a ratio of 1:4:1 [SEQ ID NO. 26], or in Formula 2, n is 1, and each X of the pentapeptide repeats is consisted of, A (Ala), G (Gly), and I (Ile) in a ratio of 1:4:1 [SEQ ID NO. 21]; K (Lys), G (Gly), and I (Ile) in a ratio of 1:4:1 [SEQ ID NO. 23]; D (Asp), G (Gly), and I (Ile) in a ratio of 1:4:1 [SEQ ID NO. 25]; or E (Glu), G (Gly), and I (Ile) in a ratio of 1:4:1 [SEQ ID NO. 27].

6 . The fusion polypeptide according to claim 4 , wherein the hydrophilic EBP is consisted of an amino acid sequence represented by Formula 2:
in Formula 2, n is 3, 6, 12 or 24, and the pentapeptide repeats correspond to SEQ ID NO. 41, SEQ ID NO. 42, SEQ ID NO. 43 or SEQ ID NO. 44 and each X of the pentapeptide repeats is consisted of A (Ala), G (Gly), and I (Ile) in a ratio of 1:4:1, or in Formula 2, n is 12, and the pentapeptide repeats correspond to [SEQ ID NO. 45] and each X of the pentapeptide repeats is consisted of E (Glu), G (Gly), and I (Ile) in a ratio of 1:4:1.

7 . The fusion polypeptide according to claim 1 , wherein the fusion polypeptide further comprises a hydrophobic elastin-based polypeptide (hydrophobic EBP) linked to the hydrophilic EBP, and
the hydrophobic EBP is consisted of an amino acid sequence represented by Formula 1 or 2 below:

Formula 1

[SEQ ID NO. 1]n;

or

Formula 2

[SEQ ID NO. 2]n, wherein

SEQ ID NO. 1 is consisted of

[VPGXG VPGXG VPGXG VPGXG VPGXG VPGXG];

SEQ ID NO. 2 is consisted of

[VPAXG VPAXG VPAXG VPAXG VPAXG VPAXG];

n is an integer of 1 or more, and represents the number of repeats of SEQ ID NO. 1 or SEQ ID NO.2; and
X is an amino acid other than proline, is selected from any natural or artificial amino acids when the pentapeptide VPGXG or VPAXG is repeated, and at least one of X is a hydrophobic or aliphatic amino acid.

8 . The fusion polypeptide according to claim 7 , wherein the hydrophobic EBP is consisted of an amino acid sequence represented by Formula 1 or 2:
in Formula 1, n is 1, and each X of the pentapeptide repeats is consisted of G (Gly), A (Ala), and F (Phe) in a ratio of 1:3:2 [SEQ ID NO. 28], or in Formula 2, n is 1, and each X of the pentapeptide repeats is consisted of, G (Gly), A (Ala), and F (Phe) in a ratio of 1:3:2 [SEQ ID NO. 29]; K (Lys), A (Ala), and F (Phe) in a ratio of 1:3:2 [SEQ ID NO. 30]; D (Asp), A (Ala), and F (Phe) in a ratio of 1:3:2 [SEQ ID NO. 31]; K (Lys) and F (Phe) in a ratio of 3:3 [SEQ ID NO. 32]; D (Asp) and F (Phe) in a ratio of 3:3 [SEQ ID NO. 33]; H (His), A (Ala), and I (Ile) in a ratio of 3:2:1 [SEQ ID NO. 34]; H (His) and G (Gly) in a ratio of 5:1 [SEQ ID NO. 35]; or G (Gly), C (Cys), and F (Phe) in a ratio of 1:3:2 [SEQ ID NO. 36].

9 . The fusion polypeptide according to claim 7 , wherein the hydrophobic EBP is consisted of an amino acid sequence represented by Formula 2:
in Formula 2, n is 12, and each X of the pentapeptide repeats is consisted of G (Gly), A (Ala), and F (Phe) in a ratio of 1:3:2 [SEQ ID NO. 46], or in Formula 2, n is 24, and each X of the pentapeptide repeats is consisted of G (Gly), A (Ala), and F (Phe) in a ratio of 1:3:2 [SEQ ID NO. 47].

10 . The fusion polypeptide according to claim 1 , wherein the fusion polypeptide is consisted of an amino acid sequence corresponding to SEQ ID NO. 48, SEQ ID NO. 49, SEQ ID NO. 50 or SEQ ID NO. 51.

11 . The fusion polypeptide according to claim 7 , wherein the fusion polypeptide is consisted of an amino acid sequence corresponding to SEQ ID NO. 52 or SEQ ID NO. 53.

12 . The fusion polypeptide according to claim 7 , wherein the fusion polypeptide forms a self-assembled nanostructure having a core-shell structure, when the hydrophobic EBP forms a core structure and the hydrophilic EBP and the VEGF receptor-specific peptide form a shell structure by a temperature stimulus.

13 . The fusion polypeptide according to claim 12 , wherein the self-assembled nanostructu

Assignees

• Iucf Hyu Erica Campus
• Univ Inje Ind Acad Coop Found

Inventors

• Lim Dong Woo
• Kang Min Jung
• Lee Jae Sang
• Park Sae-Gwang

Classifications

• A61K47/64

  Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent (peptidic linkers A61K47/65) · CPC title

• A61K38/1866

  Vascular endothelial growth factor [VEGF] · CPC title

• C07K2319/20

  containing a tag with affinity for a non-protein ligand · CPC title

• B82Y5/00

  Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title