Truncated cystine-knot proteins

• US 8,772,236 B2
• Filed: 02/05/2010
• Issued: 07/08/2014
• Est. Priority Date: 02/06/2009
• Status: Expired due to Fees

• Current Assignee: Academisch Ziekenhuis Maastricht, Pepscan Systems B.V. (Biosynth Carbosynth), Universiteit Maastricht
• Original Assignee: Academisch Ziekenhuis Maastricht, Pepscan Systems B.V. (Biosynth Carbosynth), Universiteit Maastricht
• Inventors: Timmerman, Peter, Puijk, Wouter C., Hackeng, Tilman M., Griffioen, Arjan W.
• Primary Examiner(s): Shafer, Shulamith H
• Application Number:
  US13/138,372
• Publication Number:
  US 20120231000A1
• Time in Patent Office: 1,614 Days
• Field of Search: None
• US Class Current: 514/13.3
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 15/18 Feminine contraceptives
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/52 Cytokines; Lymphokines; Interferons

Abstract

The invention relates to the fields of protein chemistry, biology and medicine. More specifically, it relates to the design and preparation of proteinmimics of members of the cystine-knot growth factor superfamily. Further, the invention relates to the use of these proteinmimics as a medicament or prophylactic agent. The invention provides proteinmimics of members of the cystine-knot growth factor superfamily, preferably for use in immunogenic and/or therapeutic compositions.

Claims

1. A proteinmimic of a member of the cystine-knot growth factor superfamily, wherein the proteinmimic comprises a motif

X0-C1-X1-C2-X2-C3-X3-C4-X4-C5-X5-C6-X6 (SEQ ID NO:

2),

wherein C1 to C6 are cysteine residues that form a cystine-knot structure in which C1 is linked to C4, C2 is linked to C5, and C3 is linked to C6,

wherein

X0 comprises KFMDVYQRSY (amino acids 1-10 of SEQ ID NO;

12),

X1 comprises HPIETLVDIFQEYPDEIEYIFKPSAVPLMR (amino acids 2-31 of SEQ ID NO;

27),

X2 comprises GGA,

X3 comprises NDEGLE (amino acids 37-42 of SEQ ID NO;

27),

X4 comprises VPTEESNITMQIMRIKPHQGQHIGEMSFLQHNK (amino acids 44-76 of SEQ ID NO;

27),

X5 comprises E, and

X6 comprises RPKKDRARQE (amino acids 90-99 of SEQ ID NO;

12),

or wherein said protein mimic consists of a sequence having at least 95% sequence identity to SEQ ID NO;

27;

wherein said protein mimic comprises the consensus sequence P[PSR]CVXXXRC2[GSTA]GCC3 (SEQ ID NO;

5) wherein at least one cysteine, other than C2 or C3, is replaced by another amino acid residue and wherein X means any amino acid, [PSR] means P or S or R and [GSTA] means G or S or T or A.

4. A pharmaceutical composition comprising the proteinmimic of claim 1 and a pharmaceutically acceptable carrier, diluent and/or excipient.

5. The pharmaceutical composition of claim 4, wherein said proteinmimic is coupled to an immunogenic carrier.

6. A method for treating a tumor-related disease wherein inhibition of angiogenesis is desired and/or treating age-related macular degeneration (AMD), the method comprising:

administering a therapeutically effective amount of the proteinmimic of claim 1 to a subject suffering from, or at risk of suffering from said disorder.

7. A method for producing antibodies against a member of the cystine-knot growth factor superfamily, comprising administering a proteinmimic according to claim 1 to a non-human animal, and obtaining antibodies against a member of the cystine-knot growth factor superfamily, which antibodies are produced by said animal.

9. The pharmaceutical composition of claim 5, wherein the immunogenic carrier is diphtheria toxin and/or keyhole limpet haemocyanin.

2. A proteinmimic member of the cystine-knot growth factor superfamily, wherein said member of the cystine-knot growth factor superfamily is placental growth factor (PLGF), and wherein said proteinmimic consists of SEQ ID NO:

14.

3. A proteinmimic of a member of the cystine-knot growth factor superfamily, wherein said member is sclerostin, and wherein said proteinmimic consists of SEQ ID NO:

31.

8. A proteinmimic of a member of the cystine-knot growth factor superfamily, wherein the proteinmimic consists of SEQ ID NO:

35.

10. A proteinmimic of human Vascular Endothelial Growth Factor (hVEGF), wherein the proteinmimic comprises a motif

X0-C1-X1-C2-X2-C3-X3-C4-X4-C5-X5-C6-X6 (SEQ ID NO:

2),

wherein C1 to C6 are cysteine residues that form a cystine-knot structure in which C1 is linked to C4, C2 is linked to C5, and C3 is linked to C6,

wherein X0 comprises KFMDVYQRSY (amino acids 1-10 of SEQ ID NO;

12), X1 comprises HPIETLVDIFQEYPDEIEYIFKPSAVPLMR (amino acids 2-31 of SEQ ID NO;

27), X2 comprises GGA, X3 comprises NDEGLE (amino acids 47-52 of SEQ ID NO;

12), X4 comprises VPTEESNITMQIMRIKPHQGQHIGEMSFLQHNK (amino acids 54-86 of SEQ ID NO;

12), X5 comprises E, and X6 comprises RPKKDRARQE (amino acids 47-52 of SEQ ID NO;

12).

11. A proteinmimic of human Vascular Endothelial Growth Factor (hVEGF), wherein the proteinmimic comprises a motif

C1-X1-C2-X2-C3-X3-C4-X4-C5-X5-C6 (SEQ ID NO:

2),

wherein C1 to C6 are cysteine residues that form a cystine-knot structure in which C1 is linked to C4, C2 is linked to C5, and C3 is linked to C6,

wherein X1 comprises HPIETLVDIFQEYPDEIEYIFKPSAVPLMR (amino acids 2-31 of SEQ ID NO;

27), X2 comprises GGA, X3 comprises NDEGLE (amino acids 37-42 of SEQ ID NO;

27), X4 comprises VPTEESNITMQIMRIKPHQGQHIGEMSFLQHNK (amino acids 44-76 of SEQ ID NO;

27), and X5 comprises E.

12. The proteinmimic of claim 11, consisting of SEQ ID NO:

27.