EP0164719A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Active site modified protease alpha-1-antitrypsin inhibitors and their production
Assignee: Novartis Vaccines and Diagnostics Inc
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Filing: 1985-06-10
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Granted: —
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Expires: 2005-06-10
Abstract
Abstract Proteinaceous compositions are provided which inhibit naturally occurring serine proteases. Particularly, an amino acid sequence analogousto human α 1 - antitrypsin is modified at the active site while maintaining protease inhibition. The methionine at the active site is substituted with an oxidatively stable amino acid, while other amino acids may also be changed, added or deleted, particularly at the termini. The proteinaceous compositions can be produced in yeast, particularly the S. carlsbergensislS. cerevisiae hybrid strain AB110 (containing plasmid pC1/1GAPATi9) deposited at the A.T.C.C. on May 9, 1984, and given Accession No. 20709.
Inventors
- 👤 Philip J. Barr
- 👤 Robert A. Hallewell
- 👤 Steven Rosenberg
- 👤 Anthony J. Brake
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-04 16:26
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/80; C12N15/81; C07; C07K; C07K14/00; C07K14/81; C07K14/8107; C07K14/811; C07K14/8121; C07K14/8125
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
58 words
A polypeptide oxidative stable serine protease inhibitor of less than about 500 amino acids, having a neutral aliphatic amino acid substituting for methionine at the inhibitory active site and, wherein said active site is flanked by amino acid sequences of at least about 5 amino acids having substantially the same amino acid sequence of human α 1 -antitrypsin.
Claim 2
Dependent Claim
58 words
A polypeptide oxidative stable serine protease inhibitor of less than about 500 amino acids, having a neutral aliphatic amino acid substituting for methionine at the inhibitory active site and, wherein said active site is flanked by amino acid sequences of at least about 5 amino acids having substantially the same amino acid sequence of human α 1 -antitrypsin.
Claim 3
Dependent Claim
12 words
An inhibitor according to Claim 1, wherein said serine protease is elastase.
Claim 4
Dependent Claim
30 words
An inhibitor according to Claim 1, wherein the amino acid sequence is substantially the amino acid sequence of the amino acids of said human α 1 -antitrypsin 310 to 390.
Claim 5
Dependent Claim
16 words
An inhibitor according to Claim 1, wherein said neutral aliphatic amino acid is valine or alanine.
Claim 6
Dependent Claim
24 words
A method for inhibiting a serine protease which comprises combining said serine protease with an inhibiting amount of an inhibitor according to Claim 1.
Claim 7
Dependent Claim
35 words
A polypeptide oxidative stable human leukocyte elastase inhibitor differing from wild-type human aI-antitrypsin by fewer than ten amino acids and having the methionine at the active site substituted with a neutral oxidatively stable amino acid.
Claim 8
Dependent Claim
35 words
A polypeptide oxidative stable human leukocyte elastase inhibitor differing from wild-type human aI-antitrypsin by fewer than ten amino acids and having the methionine at the active site substituted with a neutral oxidatively stable amino acid.
Claim 9
Dependent Claim
17 words
An inhibitor according to Claim 6, wherein said neutral oxidatively stable amino acid is valine or alanine.
Claim 10
Independent Claim
98 words
A method for producing novel serine protease inhibitors which comprises: growing a unicellular microorganism host containing a DNA sequence capable of replication and including transcriptional and translational initiation and termination signals functional in said host and structural gene under the transcriptional and translational regulation of said signals, wherein said structural gene encodes an oligopeptide sequence of at least about 20 amino acids having substantially the same sequence as human α 1 -antitrypsin and incorporating the inhibitory active site, wherein the codon for methionine at said active site is substituted by a codon expressing a neutral aliphatic amino acid.
Claim 11
Independent Claim
98 words
A method for producing novel serine protease inhibitors which comprises: growing a unicellular microorganism host containing a DNA sequence capable of replication and including transcriptional and translational initiation and termination signals functional in said host and structural gene under the transcriptional and translational regulation of said signals, wherein said structural gene encodes an oligopeptide sequence of at least about 20 amino acids having substantially the same sequence as human α 1 -antitrypsin and incorporating the inhibitory active site, wherein the codon for methionine at said active site is substituted by a codon expressing a neutral aliphatic amino acid.
Claim 12
Dependent Claim
18 words
A method according to Claim 8, wherein said DNA sequence encodes substantially the entire human α 1 -antitrypsin.
Claim 13
Dependent Claim
11 words
A method according to Claim 9, wherein said host is yeast.
Claim 14
Dependent Claim
14 words
A method according to Claim 10, wherein said yeast is a hybrid yeast strain.
Claim 15
Dependent Claim
16 words
A method according to Claim 10, wherein said yeast is an S. carlsbergensis/S. cerevisiae hybrid strain.
Claim 16
Dependent Claim
15 words
A method according to Claim 10, wherein the DNA sequence is pCl/1PHO5ATi(Val), pCl/lGAPATi(Val) or pCl/1GAPATi(Ala/Pst).
Claim 17
Dependent Claim
51 words
An S. carlsbergensis/S. cerevisiae hybrid strain obtained by crossing auxotrophic leu S. carlsbergensis yeast cells with non-complementing leu pep4 - s. cerevisiae yeast cells and selecting for an auxotrophic leu - S. carlsbergensis/S. cerevisiae hybrid strain providing for efficient expression of at least a portion of the α 1 -antitrypein gene.
Claim 18
Dependent Claim
51 words
An S. carlsbergensis/S. cerevisiae hybrid strain obtained by crossing auxotrophic leu S. carlsbergensis yeast cells with non-complementing leu pep4 - s. cerevisiae yeast cells and selecting for an auxotrophic leu - S. carlsbergensis/S. cerevisiae hybrid strain providing for efficient expression of at least a portion of the α 1 -antitrypein gene.
Claim 19
Dependent Claim
15 words
A hybrid strain according to Claim 14, which strain is AB110, A.T.C.C. Accession No. 20709.
References Cited (Backward Citations)
No citation records indexed.
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