# US 2014/0193411 A1 · Multimeric constructs

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- Number: US 2014/0193411 A1
- Document ID: US20140193411A1
- Status: Published application
- Kind: A1
- Application number: 14/170,257
- Filed: 2014-01-31
- Published: 2014-07-10
- Earliest claimed: 2007-03-12
- Assignees: Genzyme Corporation
- Applicants: Genzyme Corporation
- Inventors: Abraham Scaria; Peter Pechan; Samuel Wadsworth
- Web page: https://patentgrep.com/patent/US20140193411A1

## CPC

- C07K 16/22: against growth factors {; against growth regulators}
- A61P 3/10: for hyperglycaemia, e.g. antidiabetics
- A61P 9/10: for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
- A61P 11/06: Antiasthmatics
- A61P 19/02: for joint disorders, e.g. arthritis, arthrosis
- A61P 19/08: for bone diseases, e.g. rachitism, Paget's disease
- A61P 27/02: Ophthalmic agents
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 35/00: Antineoplastic agents
- C07K 14/475: Growth factors; Growth regulators
- C07K 14/52: Cytokines; Lymphokines; Interferons
- C07K 14/705: Receptors; Cell surface antigens; Cell surface determinants {(tumour specific antigens C07K14/4748)}
- C12P 21/00: Preparation of peptides or proteins (single cell protein C12N1/00)
- 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)
- C07K 2319/30: Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
- C07K 2319/32: fusions with soluble part of a cell surface receptor, "decoy receptors"
- C07K 2319/735: containing a domain for self-assembly, e.g. a viral coat protein (includes phage display)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## Abstract

Multimeric fusion proteins of an Ig-like domain of Flt-1 are rendered functional by inclusion of a linker moiety. Vectors encoding the fusion proteins and host cells expressing the fusion proteins can be used therapeutically to block neovascularization in individuals with pathological conditions related to neovascularization. Such conditions include age-related macular degeneration, cancer, psoriasis, proliferative diabetic retinopathy, asthma, uveitis, osteoarthritis, and rheumatoid arthritis. The same means of multimerization used for an Iglike domain of Flt-1, i.e., a linker and a multimerization domain, can be used for other polypeptides, including extracellular receptors, antibody variable regions, cytokines, chemokines, and growth factors.

## Claims

### Claim 1

A method of inhibiting VEGF activity in a cell comprising delivering to the cell a nucleic acid molecule that encodes a fusion protein of the formula X-Y-Z, wherein
X comprises the Ig-like domain 2 of VEGF-R1 but lacks the Ig-like domains 1 and 3 of a VEGF receptor, with said Ig-like domain 2 of VEGF-R1 being covalently linked to moiety Z via moiety Y;
Y is selected from the group consisting of gly9 (SEQ ID NO: 27), glu9 (SEQ ID NO: 28), ser9 (SEQ ID NO: 29), gly5-cys-pro2-cys (SEQ ID NO: 30), (gly4ser)3 (SEQ ID NO: 31), Ser-Cys-Val-Pro-Leu-Met-Arg-Cys-Gly-Gly-Cys-Cys-Asn (SEQ ID NO: 32), Pro-Ser-Cys-Val-Pro-Leu-Met-Arg-Cys-Gly-Gly-Cys-Cys-Asn (SEQ ID NO: 13), Gly-Asp-Leu-Ile-Tyr-Arg-Asn-Gln-Lys (SEQ ID NO: 26), and Gly9-Pro-Ser-Cys-Val-Pro-Leu-Met-Arg-Cys-Gly-Gly-Cys-Cys-Asn (SEQ ID NO: 34), and
Z is a CH3 region of an IgG heavy chain molecule or an Fc portion of an antibody molecule,
whereby an effective amount of said fusion protein is expressed in the cell thereby inhibiting VEGF activity.

### Claim 2 (depends on 1)

The method of claim 1, wherein the fusion protein comprises a sequence selected from the group consisting of SEQ ID NO: 2, 8, 21, 23, and 25.

### Claim 3 (depends on 1)

The method of claim 1, wherein the nucleic acid is delivered in vivo.

### Claim 4 (depends on 1)

The method of claim 1, wherein Z is an Fc portion of an antibody molecule.

### Claim 5 (depends on 1)

The method of claim 1, wherein X consists of the Ig-like domain 2 of VEGF-R1 (Flt-1).

### Claim 6

A method of inhibiting VEGF activity in a mammal comprising delivering to the mammal a therapeutically effective amount of a fusion protein of the formula X-Y-Z, wherein X comprises the Ig-like domain 2 of VEGF-R1 but lacks the Ig-like domains 1 and 3 of a VEGF receptor, with said Ig-like domain 2 of VEGF-R1 being covalently linked to moiety Z via moiety Y;
Y is selected from the group consisting of gly9 (SEQ ID NO: 27), glu9 (SEQ ID NO: 28), ser9 (SEQ ID NO: 29), gly5-cys-pro2-cys (SEQ ID NO: 30), (gly4ser)3 (SEQ ID NO: 31), Ser-Cys-Val-Pro-Leu-Met-Arg-Cys-Gly-Gly-Cys-Cys-Asn (SEQ ID NO: 32), Pro-Ser-Cys-Val-Pro-Leu-Met-Arg-Cys-Gly-Gly-Cys-Cys-Asn (SEQ ID NO: 13), Gly-Asp-Leu-Ile-Tyr-Arg-Asn-Gln-Lys (SEQ ID NO: 26), and Gly9-Pro-Ser-Cys-Val-Pro-Leu-Met-Arg-Cys-Gly-Gly-Cys-Cys-Asn (SEQ ID NO: 34), and
Z is a CH3 region of an IgG heavy chain molecule or an Fc portion of an antibody molecule,
thereby inhibiting VEGF activity.

### Claim 7 (depends on 6)

The method of claim 6, wherein X consists of the IgG-like domain 2 of VEGF-R1 (FLT-1).

### Claim 8 (depends on 6)

The method of claim 6, wherein said fusion protein is capable of binding to VEGF.

### Claim 9 (depends on 6)

The method of claim 6, wherein Z is an IgG1 CH3 region.

### Claim 10 (depends on 6)

The method of claim 6, wherein Z is an IgG2 CH3 region.

### Claim 11 (depends on 6)

The method of claim 6, wherein the Fc is an IgG Fc.

### Claim 12 (depends on 11)

The method of claim 11, wherein the IgG Fc is IgG1 Fc.

### Claim 13 (depends on 6)

The method of claim 6, wherein the mammal has wet age-related macular degeneration or proliferative diabetic retinopathy.

### Claim 14 (depends on 6)

The method of claim 6, wherein the mammal has cancer.

### Claim 15 (depends on 6)

The method of claim 6, wherein the mammal has rheumatoid arthritis.

### Claim 16 (depends on 6)

The method of claim 6, wherein the mammal has asthma.

### Claim 17 (depends on 6)

The method of claim 6, wherein the mammal has osteoarthritis.

### Claim 18 (depends on 6)

The method of claim 6, wherein neovascularization is inhibited.