INHIBITORS OF TYPE 2 VASCULAR ENDOTHELIAL GROWTH FACTOR RECEPTORS

• US 20140179896A1
• Filed: 11/04/2013
• Published: 06/26/2014
• Est. Priority Date: 12/05/2003
• Status: Active Grant

• Current Assignee: Bristol-Myers Squibb Company
• Original Assignee: Bristol-Myers Squibb Company
• Inventors: CHEN, Yan, WRIGHT, Martin C., LIM, Ai Ching, GETMANOVA, Elena, HARRIS, Alan S., GOKEMEIJER, Jochem, SUN, Lin, WITTEKIND, Michael
• Granted Patent: US 9,328,157 B2
• Time in Patent Office: Days
• US Class Current: 530/300
• CPC Class Codes: A61K 2039/505 comprising antibodies
• 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: A61K 38/39 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 27/06 Antiglaucoma agents or miotics
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 35/04 : specific for metastasis
• CPC Class Codes: A61P 37/00 : Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 : Immunomodulators
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• 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: C07H 21/04 : with deoxyribosyl as saccharide radical
• CPC Class Codes: C07K 14/4703 : Inhibitors; Suppressors
• CPC Class Codes: C07K 14/52 : Cytokines; Lymphokines; Interferons
• CPC Class Codes: C07K 14/78 : Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
• CPC Class Codes: C07K 16/2863 : against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 16/30 : from tumour cells
• CPC Class Codes: C07K 19/00 : Hybrid peptides , i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
• CPC Class Codes: C07K 2317/34 : Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/73 : Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2318/20 : Antigen-binding scaffold molecules wherein the scaffold is not an immunoglobulin variable region or antibody mimetics
• CPC Class Codes: C07K 2319/21 : containing a His-tag
• CPC Class Codes: C07K 2319/23 : containing a GST-tag
• CPC Class Codes: C07K 2319/30 : Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 2319/31 : fusions, other than Fc, for prolonged plasma life, e.g. albumin
• CPC Class Codes: C07K 2319/41 : containing a Myc-tag
• CPC Class Codes: C07K 2319/42 : containing a HA(hemagglutinin)-tag
• CPC Class Codes: C07K 2319/43 : containing a FLAG-tag
• CPC Class Codes: C12N 15/09 : Recombinant DNA-technology
• CPC Class Codes: C12N 15/1062 : mRNA-Display, e.g. polypeptide and encoding template are connected covalently
• CPC Class Codes: C40B 40/08 : Libraries containing RNA or DNA which encodes proteins, e.g. gene libraries
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof

Abstract

The present disclosure relates to novel vascular endothelial growth factor receptor (VEGFR)-binding polypeptides and methods for using these polypeptides to inhibit biological activities mediated by vascular endothelial growth factors (VEGFs). The present disclosure also provides various improvements relating to single domain binding polypeptides.

Claims

1-6. -6. (canceled)

7. A polypeptide comprising a fibronectin type III tenth (¹⁰Fn3) domain, wherein the ¹⁰Fn3 domain (i) comprises a modified amino acid sequence in one or more of the AB, BC, CD, DE, EF and FG loops relative to the wild-type ¹⁰Fn3 domain set forth in SEQ ID NO:

5, (ii) binds to a target molecule not bound by the wild-type ¹⁰Fn3 domain, and (iii) comprises a C-terminal tail comprising the sequence EIDKPXQ, wherein X is C or S.

8. The polypeptide of claim 7, wherein the lysine corresponding to amino acid position 8 of SEQ ID NO:

5 is substituted with glutamine.

9. An isolated nucleic acid encoding the polypeptide of claim 7.

10. A polypeptide comprising a fibronectin type III tenth (¹⁰Fn3) domain, wherein the ¹⁰Fn3 domain (i) has a deletion or mutation of 1 to 8 amino acids in the N-terminus of the wild-type ¹⁰Fn3 domain set forth in SEQ ID NO:

5, (ii) comprises a modified amino acid sequence in one or more of the AB, BC, CD, DE, EF and FG loops relative to the wild-type ¹⁰Fn3 domain, and (iii) binds to a target molecule not bound by the wild-type ¹⁰Fn3 domain.

11. The polypeptide of claim 10, wherein the polypeptide has a deletion of 1 to 8 amino acids at the N-terminus of the wild-type ¹⁰Fn3 domain set forth in SEQ ID NO:

5.

12. The polypeptide of claim 11, wherein the polypeptide has as deletion of 8 amino acids at the N-terminus of the wild-type ¹⁰Fn3 domain set forth in SEQ ID NO:

5.

13. The polypeptide of claim 10, wherein one or two amino acid residues are added to the N-terminus of the ¹⁰Fn3 domain.

14. The polypeptide of claim 11, wherein a methionine and/or glycine residue is added back to the N-terminus of the ¹⁰Fn3 domain.

15. The polypeptide of claim 10, wherein the lysine corresponding to amino acid position 8 of SEQ ID NO:

5 is substituted with glutamine.

16. The polypeptide of claim 10, wherein the ¹⁰Fn3 domain comprises a C-terminal tail of 1 to 7 amino acids.

17. The polypeptide of claim 10, wherein binding of the polypeptide to the target molecule is increased in the absence of the N-terminal VSDVPRDL sequence relative to a polypeptide which is identical in sequence but comprises the N-terminal VSDVPRDL sequence.

18. An isolated nucleic acid encoding the polypeptide of claim 10.

24. The polypeptide of claim 17, wherein the target molecule and the homology from the second mammal shares at least 80% identity across the entire protein sequence.

19. A polypeptide comprising a fibronectin type III tenth (¹⁰Fn3) domain, wherein the ¹⁰Fn3 domain (i) comprises a modified amino acid sequence in one or more of the AB, BC, CD, DE, EF and FG loops relative to the wild-type ¹⁰Fn3 domain, (ii) binds to a target molecule not bound by the wild-type ¹⁰Fn3 domain;

and (iii) binds to the target molecule from a first mammal and to a homolog thereof from a second mammal.

20. The polypeptide of claim 19, wherein the first mammal is a human.

21. The polypeptide of claim 20, wherein the second mammal is a mammal used in preclinical testing.

22. The polypeptide of claim 21, wherein the second mammal is a mouse, rat, guinea pig, dog, or non-human primate.

23. The polypeptide of claim 20, wherein the target molecule and the homolog from the second mammal shares at least 80% identity across a region of at least 50 amino acids.

25. The polypeptide of claim 20, wherein the ¹⁰Fn3 domain binds to both the human target molecule and homolog thereof with a dissociation constant of less than 1×

10<sup>−

6</sup>M.

26. An isolated nucleic acid encoding the polypeptide of claim 19.