EP0320148A1
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Analogs of fibroblast growth factor
Assignee: Amgen Inc
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Filing: 1988-11-23
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Granted: —
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Expires: 2008-11-23
Abstract
Abstract Disclosed are recombinant basic fibroblast growth factor analogs possessing part or all of the primary structural conformation and one or more of the biological properties of a mammalian (e.g., human) basic fibroblast growth factor, wherein at least one of the cysteine residues of the naturally occurring basic fibroblast growth factor is replaced with a residue of a different amino acid. Also disclosed is a process for producing such analogs wherein a host cell is transformed or transfected with an exogenous DNA sequence encoding for the basic fibroblast growth factor analogs. Purification methods for the analogs are also disclosed as well as in vivo applications. Biologically active analogs of basic fibroblast growth factor are more stable and facilitate purification of basic fibroblast growth factor.
Inventors
- 👤 Tsutomu Arakawa
- 👤 Gary M. Fox
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-03 16:09
CPC Classifications
C; C07; C07H; C07H15/00; C07H15/02; C07H15/12; C07K; C07K14/00; C07K14/435; C07K14/475; C07K14/50; C07K14/503; A; A61; A61P; A61P17/00; A61P43/00; A61K; A61K38/00
Claims Analysis
Total Claims: 33
Claim 1
Independent Claim
57 words
An analog of basic fibroblast growth factor which differs from naturally occurring basic fibroblast growth factor in terms of the identity and/or location of one of more amino acid residues, wherein at least one of the cysteine residues of said naturally occurring basic fibroblast growth factor is is replaced by a residue of a different amino acid.
Claim 2
Dependent Claim
57 words
An analog of basic fibroblast growth factor which differs from naturally occurring basic fibroblast growth factor in terms of the identity and/or location of one of more amino acid residues, wherein at least one of the cysteine residues of said naturally occurring basic fibroblast growth factor is is replaced by a residue of a different amino acid.
Claim 3
Dependent Claim
23 words
The analog of Claim 1 wherein said different amino acid is selected from the group consisting of serine, alanine, aspartic acid and asparagine.
Claim 4
Dependent Claim
12 words
The analog of Claim 2 wherein, said different amino acid is serine.
Claim 5
Dependent Claim
23 words
The analog of Claim 1 wherein at least one of said replaced cysteine residues is a cysteine residue existing as a free sulfhydryl.
Claim 6
Dependent Claim
21 words
The analog of Claim 1 wherein two of said cysteine residues are replaced by a residue of a different amino acid.
Claim 7
Dependent Claim
19 words
The analog of Claim 5 wherein two of said replaced cysteine residues are cysteine residues existing as free sulfhydryls.
Claim 8
Dependent Claim
46 words
An analog of basic fibroblast growth factor having the amino acid sequence set forth in Table VI and allelic variants thereof, wherein at least one of the cysteine residues at positions 26, 70, 88, and 93, is replaced by a residue of a different amino acid.
Claim 9
Dependent Claim
46 words
An analog of basic fibroblast growth factor having the amino acid sequence set forth in Table VI and allelic variants thereof, wherein at least one of the cysteine residues at positions 26, 70, 88, and 93, is replaced by a residue of a different amino acid.
Claim 10
Dependent Claim
30 words
The analog of Claim 7 wherein at least one terminal amino acid residue is deleted while said analog substantially retains the biological activity of naturally occurring basic fibroblast growth factor.
Claim 11
Dependent Claim
23 words
The analog of Claim 7 wherein said different amino acid is selected from the groups consisting of serine, alanine, aspartic acid, and asparagine.
Claim 12
Dependent Claim
12 words
The analog of Claim 8 wherein said different amino acid is serine.
Claim 13
Dependent Claim
29 words
The analog of Claim 7 wherein at least one of said replaced cysteine residues is selected from the group consisting of cysteines at amino acid positions 70 and 88.
Claim 14
Dependent Claim
21 words
The analog of Claim 7 wherein two of said cysteine residues is replaced by a residue of a different amino acid.
Claim 15
Dependent Claim
19 words
The analog of Claim 11 wherein said replaced amino acids comprise cysteines at amino acid positions 70 and 88.
Claim 16
Dependent Claim
19 words
The analog of Claim 11 wherein said replaced cysteine residues comprise cysteines at amino acid positions 26 and 93.
Claim 17
Dependent Claim
21 words
The analog of Claim 12 wherein said replaced cysteine residues comprise cysteines at amino acid positions 26, 70, 88, and 93.
Claim 18
Dependent Claim
21 words
A DNA sequence encoding for procaryotic or eucaryotic expression of an analog of a basic fibroblast growth factor of Claim 7.
Claim 19
Dependent Claim
18 words
The DNA sequence of Claim 16 wherein said DNA sequence is modified from human fibroblast growth factor gene.
Claim 20
Dependent Claim
20 words
The DNA sequence of Claim 17 wherein said human basic fibroblast growth factor gene is modified by oligonucleotide site-directed mutagenesis.
Claim 21
Dependent Claim
23 words
The DNA sequence of Claim 18 wherein said human basic fibroblast growth factor is modified from a bovine basic fibroblast growth factor gene.
Claim 22
Dependent Claim
19 words
The DNA sequence of Claim 19 wherein said bovine fibroblast growth factor gene is modified by oligonucleotide site-directed mutagenesis.
Claim 23
Dependent Claim
23 words
A pharmaceutical composition comprising a therapeutically effective amount of a basic fibroblast growth factor analog according to Claim 1 and pharmaceutically acceptable adjuvants.
Claim 24
Dependent Claim
26 words
A method for treating a wound comprising administering to said wound a therapeutically effective amount of a basic fibroblast growth factor analog according to Claim 1.
Claim 25
Dependent Claim
12 words
The method of Claim 22 wherein said wound is a surface wound.
Claim 26
Dependent Claim
12 words
The method of Claim 22 wherein said wound is a surgical wound.
Claim 27
Dependent Claim
14 words
The method of Claim 22 wherein said wound includes a bone fracture or defect.
Claim 28
Dependent Claim
12 words
The method of Claim 22 wherein said wound includes a damaged nerve.
Claim 29
Dependent Claim
24 words
A method for generating tissue and/or organs comprising administering a therapeutically effective amount of a basic fibroblast growth factor analog according to Claim 1.
Claim 30
Dependent Claim
33 words
A procaryotic or eucaryotic host cell transformed or transfected with DNA according to Claim 16 in a manner allowing the host cell to express a basic fibroblast growth factor analog of Claim 7.
Claim 31
Dependent Claim
56 words
A method of producing a purified and isolated basic fibroblast growth factor analog comprising the steps of: transfecting or transforming host cells with DNA according to Claim 16; culturing the transfected or transformed host cells to allow the host cells to express a basic fibroblast growth factor analog; and, isolating said basic fibroblast growth factor analog.
Claim 32
Dependent Claim
31 words
A method for the purification of a recombinant basic fibroblast growth factor analog according to Claim 7 comprising subjecting a supernatant containing basic fibroblast growth factor analog to non heparin chromatography.
Claim 33
Dependent Claim
44 words
A method for the purification of a recombinant basic fibroblast growth factor analog according to Claim 7 comprising the steps of: solubilizing the inclusion bodies from host cell cultures containing basic fibroblast growth factor; and, subjecting the solubilized inclusion bodies to non heparin chromatography.
References Cited (Backward Citations)
No citation records indexed.
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