Fibroblast growth factor mutants having improved functional half-life and methods of their use

Patent metadata

Field	Value
Publication number	US-10000540-B2
Application number	US-201615295833-A
Country	US
Kind code	B2
Filing date	Oct 17, 2016
Priority date	May 20, 2009
Publication date	Jun 19, 2018
Grant date	Jun 19, 2018

Abstract

Mutant fibroblast growth factor (FGF) proteins having a polypeptide sequence with a high sequence identity to proteins encoded by members of the Fgf-1 subfamily of genes from a mammalian species, such as human, and with a specific amino acid substitution of an alanine at a position corresponding to amino acid position 66 of human FGF-1 with a cysteine and/or a specific amino acid substitution of a phenylalanine at a position corresponding to amino acid position 132 of human FGF-1 with a tryptophan (based on the 140 amino acid numbering scheme of human FGF-1) are provided. Other amino acid mutations or substitutions may be combined. Polynucleotide sequences encoding the mutant FGF proteins and host cells containing such polynucleotide sequences are provided. Methods of administering a mutant FGF protein to an individual to treat an ischemic condition or disease or a wound or tissue injury are also provided.

First claim

That which is claimed:

1. A composition comprising a mutant fibroblast growth factor (FGF) protein having a polypeptide sequence that is at least 90% identical to the wild-type human FGF-1 protein having SEQ ID NO: 1, the phenylalanine at an amino acid position of the mutant FGF polypeptide sequence corresponding to amino acid position 132 of SEQ ID NO: 1 being substituted with tryptophan, the mutant FGF protein have a greater functional half-life than SEQ ID NO: 1.

2. The composition of claim 1 , wherein the mutant FGF protein comprises SEQ ID NO: 7.

3. The composition of claim 1 , wherein the mutant FGF protein consists of SEQ ID NO: 7.

4. The composition of claim 1 , wherein the mutant FGF protein has a polypeptide sequence that is at least 95% identical to the wild-type human FGF-1 protein having SEQ ID NO: 1.

5. The composition of claim 1 , wherein the mutant FGF protein is contained in a pharmaceutical composition with a pharmaceutically acceptable carrier.

6. The composition of claim 1 , wherein the mutant FGF protein includes at least one additional mutation selected from: Leu44→Phe, Met 67→Ile, Leu73→Val, Val109→Leu, Leu111→Ile and Cys117→Val.

7. The composition of claim 1 , wherein the mutant FGF protein includes at least one additional mutation selected from: Ile42→Cys, Cys83→Ile, Ile130→Cys, Phe22→Tyr, Tyr64→Phe, and Phe108→Tyr.

8. The composition of claim 1 , wherein the mutant FGF protein includes at least one additional mutation selected from:
(a) Lys12 substituted with Cys, Thr, or, Val;
(b) Leu46 substituted with Val;
(c) Glu87 substituted with Val;
(d) Asn95 substituted with Val; and
(e) Pro134 substituted with Cys, Thr, or Val.

9. A method of triggering proliferation of cells expressing at least one fibroblast growth factor receptor (FGFR) in a patient, the method comprising:
administering to a patient having a wound, tissue damage, and/or an ischemic condition a therapeutically effective amount of composition comprising a mutant fibroblast growth factor (FGF) protein having a polypeptide sequence that is at least 90% identical to the wild-type human FGF-1 protein having SEQ ID NO: 1, the phenylalanine at an amino acid position of the mutant FGF polypeptide sequence corresponding to amino acid position 132 of SEQ ID NO: 1 being substituted with tryptophan, the mutant FGF protein have a greater functional half-life than SEQ ID NO: 1, wherein the mutant FGF protein triggers proliferation of cells expressing at least one FGFR.

10. The method of claim 9 , wherein the mutant FGF protein comprises SEQ ID NO:7.

11. The method of claim 9 , wherein the mutant FGF protein consists of SEQ ID NO: 7.

12. The method of claim 9 , wherein the mutant FGF protein has a polypeptide sequence that is at least 95% identical to the wild-type human FGF-1 protein having SEQ ID NO: 1.

13. The method of claim 9 , wherein the mutant FGF protein is contained in a pharmaceutical composition with a pharmaceutically acceptable carrier.

14. The method of claim 9 , wherein the mutant FGF protein includes at least one additional mutation selected from: Leu44→Phe, Met 67→Ile, Leu73→Val, Val109→Leu, Leu111→Ile and Cys117→Val.

15. The method of claim 9 , wherein the mutant FGF protein includes at least one additional mutation selected from: Ile42→Cys, Cys83→Ile, Ile130→Cys, Phe22→Tyr, Tyr64→Phe, and Phe108→Tyr.

16. The method of claim 9 , wherein the mutant FGF protein includes at least one additional mutation selected from:
(a) Lys12 substituted with Cys, Thr, or, Val;
(b) Leu46 substituted with Val;
(c) Glu87 substituted with Val;
(d) Asn95 substituted with Val; and
(e) Pro134 substituted with Cys, Thr, or Val.

Assignees

• Univ Florida State Res Found

Inventors

• Blaber Michael
• Lee Jihun

Classifications

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P17/02

  for treating wounds, ulcers, burns, scars, keloids, or the like · CPC title

• C07H21/00

  Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids · CPC title

• A61K38/18

  Growth factors; Growth regulators · CPC title

• A61K38/00

  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 title