METHODS AND COMPOSITIONS FOR BINDING VEGF

• US 20190002546A1
• Filed: 05/21/2018
• Published: 01/03/2019
• Est. Priority Date: 11/19/2015
• Status: Active Application

• Current Assignee: Zhuhai Tairuishang Biopharm, Ltd.
• Original Assignee: Zhuhai Tairuishang Biopharm, Ltd.
• Inventors: Li, Ronghao, Wu, John Jun
• Application Number:
  US15/985,541
• Publication Number:
  US 20190002546A1
• Time in Patent Office: Days
• CPC Class Codes: A61K 38/1866 Vascular endothelial growth factor [VEGF]
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61P 27/00 Drugs for disorders of the senses
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/71 for growth factors; for growth regulators
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2319/01 containing a localisation/targetting motif
• 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 2319/32 fusions with soluble part of a cell surface receptor, "decoy receptors"
• CPC Class Codes: C07K 2319/70 containing domain for protein-protein interaction
• CPC Class Codes: C07K 2319/735 containing a domain for self-assembly, e.g. a viral coat protein (includes phage display)
• CPC Class Codes: C07K 2319/74 containing a fusion for binding to a cell surface receptor
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins

Abstract

The present invention relates to fusion polypeptide compositions comprising immunoglobulin-type-2 binding domains of Vascular Endothelial Growth Factor Receptor, isolated nucleic acids encoding the compositions and vectors and host cells containing the same, and methods of using such compositions in treatment of diseases, disorders, and conditions.

Claims

1. A fusion polypeptide comprising two or more VEGF receptor immunoglobulin-like-type 2 domains fused with a dimerization polypeptide, wherein at least one of said two or more VEGF receptor immunoglobulin-like-type 2 domains is fused to an N-terminus of said dimerization polypeptide and at least another of said two or more VEGF receptor immunoglobulin-like-type 2 domains is fused to a C-terminus of said dimerization polypeptide.

3. The fusion polypeptide of claim 1, wherein said dimerization polypeptide is a fragment crystallizable (Fc) domain.

4. The fusion polypeptide of claim 1, wherein said dimerization domain comprises a first cysteine residue capable of forming a disulfide bond to a second cysteine residue.

5. The fusion polypeptide of claim 1, further comprising at least one hinge region between said dimerization polypeptide and said two or more VEGF receptor immunoglobulin-like-type 2 domains.

6. The fusion polypeptide of claim 1, wherein said two or more VEGF receptor immunoglobulin-like-type 2 domains:

are each at least 80% identical to a human VEGF receptor immunoglobulin-like-type 2 domain selected from the group consisting of SEQ ID NOs 4-6;

are each at least 90% identical to a human VEGF receptor immunoglobulin-like-type 2 domain selected from the group consisting of SEQ ID NOs 4-6;

are each at least 95% identical to a human VEGF receptor immunoglobulin-like-type 2 domain selected from the group consisting of SEQ ID NOs 4-6;

are each a human VEGF receptor immunoglobulin-like-type 2 domain selected from the group consisting of SEQ ID NOs 4-6;

are each SEQ ID NO;

4;

are each SEQ ID NO;

5;

or

are each SEQ ID NO;

6.

13. The fusion polypeptide of claim 1, wherein said two or more VEGF receptor immunoglobulin-like-type 2 domains are not identical.

14. The fusion polypeptide of claim 13, wherein said two or more VEGF receptor immunoglobulin-like-type 2 domains are at least two distinct human VEGF receptor immunoglobulin-like-type 2 domains.

15. The fusion polypeptide of claim 14, wherein said at least two distinct human VEGF receptor immunoglobulin-like-type 2 domains are selected from the group consisting of SEQ ID NOs:

4-6.

16. The fusion polypeptide of claim 1, wherein said fusion polypeptide substantially lacks a VEGF receptor immunoglobulin-like-type 3 domain selected from the group consisting of SEQ ID NOs 1-3.

17. The fusion polypeptide of claim 1, wherein said fusion polypeptide comprises no more than 60 amino acids of a VEGF receptor immunoglobulin-like-type 3 domain selected from the group consisting of SEQ ID NOs 1-3.

18. The fusion polypeptide of claim 16, wherein said VEGF receptor immunoglobulin-like-type 3 domain is:

at least 80% identical to a human VEGF receptor immunoglobulin-like-type 3 domain selected from the group consisting of SEQ ID NOs 1-3;

at least 90% identical to a human VEGF receptor immunoglobulin-like-type 3 domain selected from the group consisting of SEQ ID NOs 1-3;

at least 95% identical to a human VEGF receptor immunoglobulin-like-type 3 domain selected from the group consisting of SEQ ID NOs 1-3;

SEQ ID NO;

1;

SEQ ID NO;

2;

or

SEQ ID NO;

3.

24. The fusion polypeptide of claim 1, wherein said fusion polypeptide does not comprise SEQ ID NO:

8.

25. The fusion polypeptide of claim 1, wherein a homodimer of said fusion polypeptide exhibits high-affinity binding to VEGF.

26. The fusion polypeptide of claim 25, wherein said homodimer binds to VEGF with a higher affinity than the polypeptide shown in SEQ ID NO:

9.

27. The fusion polypeptide of claim 25, wherein said homodimer binds to VEGF with a higher affinity than a human VEGF receptor.

29. An isolated polynucleotide molecule encoding said fusion polypeptide of claim 1.

30. The isolated polynucleotide molecule of claim 29, wherein said polynucleotide comprises SEQ ID NO:

7.

37. A method for inhibiting angiogenesis in a subject in need thereof comprising:

administering to said subject in need thereof a therapeutically effective amount of a homodimer of said fusion polypeptide of claim 1.

38. The method of claim 37, wherein said administering is effected by a local administration or a systemic administration to said subject.

39. The method of claim 38, wherein said administration is to an eye of said subject.

40. The method of claim 39, wherein said administration is to a tumor tissue of said subject.

41. The method of claim 39, wherein said administration is intravenous injection, intraperitoneal injection, or intraperitoneal injection.

44. The method of claim 37, wherein said angiogenesis is a manifestation of a condition selected from the group consisting of age-related macular degeneration, diabetic retinopathy, choroidal neovascularization, cystoid macular edema, diabetic macular edema, retinal vascular occlusion, corneal neovascularization, corneal transplantation, neovascular glaucoma, pterygium chronic conjunctivitis, angiogenesis related therapy failure such as laser coagulation, and surgical retinal transplantation.

45. The method of claim 44, wherein said condition is AMD.

46. The method of claim 44, wherein said condition is diabetic retinopathy.

47. The method of claim 44, wherein said administering results in one or more improved symptoms of said condition, wherein said symptoms are selected from the group consisting of a decrease in mean choroidal neovascularization (CNV) leakage, improved mean visual acuity, a reduction in mean foveal retinal thickness, a reduction in mean macular size, and a reduction in mean lesion size.

48. The method of claim 47, wherein said one or more improved symptoms of said condition remains improved for at least 1 month following said administration.

49. The method of claim 47, wherein said homodimer is administered by intravitreal injection at an amount from about 1 mg to about 3 mg

50. The method of claim 49, wherein said homodimer is administered by intravitreal injection of an amount of about 2 mg.

51. The method of claim 37, wherein said angiogenesis is a manifestation of a tumor.

53. The method of claim 51, wherein said fusion polypeptide is administered by an intravenous injection comprising an amount of from about 0.1 to about 30 mg/kg, or from about 1 to about 8 mg/kg.

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