Human bikunin

• US 7,019,123 B2
• Filed: 10/10/2001
• Issued: 03/28/2006
• Est. Priority Date: 03/11/1996
• Status: Expired due to Fees

• Current Assignee: Bayer Corporation (Bayer AG)
• Original Assignee: Bayer Corporation (Bayer AG)
• Inventors: Tamburini, Paul P., Davis, Gary, Muller, Daniel K., Delaria, Katherine A., Marlor, Christopher W.
• Primary Examiner(s): Weber, Jon
• Assistant Examiner(s): Liu, Samuel W.
• Application Number:
  US09/974,026
• Publication Number:
  US 20030194398A1
• Time in Patent Office: 1,630 Days
• Field of Search: 536/23.1, 514/8, 514/44, 435/320.1, 435/440, 435/195, 435/69.1
• US Class Current: 536/23.1
• CPC Class Codes: 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
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 7/04 Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/8114 Kunitz type inhibitors
• CPC Class Codes: C07K 14/8117 Bovine/basic pancreatic trypsin inhibitor (BPTI, aprotinin)

Abstract

The instant invention provides for proteins, polypeptides, nucleic acid sequences, constructs, expression vectors, host cells, pharmaceutical compositions of, and methods for using human placental bikunin, serine protease inhibitor domains, and fragments thereof

Claims

1. An isolated nucleic acid sequence, wherein the nucleic acid encodes a protein having serine protease inhibitory activity, comprising one of the following amino acid sequences:

(SEQ ID NO;

52)ADRERSIHDF CLVSKVVGRC RASMPRWWYNVTDGSCQLFV YGGCDGNSNN50 YLTKEECLKK CATVTENATG DLATSRNAADSSVPSAPRRQ DSEDHSSDMF100 NYEEYCTANA VTGPCRASFP RWYFDVERNSCNNFIYGGCR GNKNSYRSEE150 ACMLRCFRQQ ENPPLPLGSK;

170 (SEQ ID NO;

49) 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

MAQLCGLRRSRAFLALL GSLLLSGVLA−

1 ADRERSIHDF CLVSKVVGRC RASMPRWWYNVTDGSCQLFV YGGCDGNSNN50 YLTKEECLKK CATVTENATG DLATSRNAADSSVPSAPRRQ DSEDHSSDMF100 NYEEYCTANA VTGPCRASFP RWYFDVERNSCNNFIYGGCR GNKNSYRSEE150 ACMLRCFRQQ ENPPLPLGSK VVVLAGLFVMVLILFLGASM VYLIRVARRN200 QERALRTVWS SGDDKEQLVK NTYVL;

225 (SEQ ID NO;

71)ADRERSIHDF CLVSKVVGRC RASMPRWWYNVTDGSCQLFV YGGCDGNSNN50 YLTKEECLKK CATVTENATG DLATSRNAADSSVPSAPRRQ DSEDHSSDMF100 NYEEYCTANA VTGPCRASFP RWYFDVERNSCNNFIYGGCR GNKNSYRSEE150 ACMLRCFRQQ ENPPLPLGSK VVVLAGLFVMVLILFLGASM VYLIRVARRN200 QERALRTVWS SGDDKEQLVK NTYVL;

225 (SEQ ID NO;

2)AGSFLAWL GSLLLSGVLA−

1 ADRERSIHDF CLVSKVVGRC RASMPRWWYNVTDGSCQLFV YGGCDGNSNN50 YLTKEECLKK CATVTENATG DLATSRNAADSSVPSAPRRQ DSEDHSSDMF100 NYEEYCTANA VTGPCRASFP RWYFDVERNSCNNFIYGGCR GNKNSYRSEE150 ACMLRCFRQQ ENPPLPLGSK VVVLAGAVS179 (SEQ ID NO;

45) 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

MLRAEADGVSRLL GSLLLSGVLA−

1 ADRERSIHDF CLVSKVVGRC RASMPRWWYNVTDGSCQLFV YGGCDGNSNN50 YLTKEECLKK CATVTENATG DLATSRNAADSSVPSAPRRQ DSEDHSSDMF100 NYEEYCTANA VTGPCRASFP RWYFDVERNSCNNFIYGGCR GNKNSYRSEE150 ACMLRCFRQQ ENPPLPLGSK VVVLAGLFVMVLILFLGASM VYLIRVARRN200 QERALRTVWS SGDDKEQLVK NTYVL;

225 (SEQ ID NO;

47) 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

MAQLCGLRRSRAFLALL GSLLLSGVLA−

1 ADRERSIHDF CLVSKVVGRC RASMPRWWYNVTDGSCQLFV YGGCDGNSNN50 YLTKEECLKK CATVTENATG DLATSRNAADSSVPSAPRRQ DSEDHSSDMF100 NYEEYCTANA VTGPCRASFP RWYFDVERNSCNNFIYGGCR GNKNSYRSEE150 ACMLRCFRQQ ENPPLPLGSK VVVLAGLFVMVLILFLGASM VYLIRVARRN200 QERALRTVWS FGD;

213 (SEQ ID NO;

70)ADRERSIHDF CLVSKVVGRC RASMPRWWYNVTDGSCQLFV YGGCDGNSNN50 YLTKEECLKK CATVTENATG DLATSRNAADSSVPSAPRRQ DSEDHSSDMF100 NYEEYCTANA VTGPCRASFP RWYFDVERNSCNNFIYGGCR GNKNSYRSEE150 ACMLRCFRQQ ENPPLPLGSK VVVLAGLFVMVLILFLGASM VYLIRVARRN200 QERALRTVWS FGD;

213 (SEQ ID NO;

4) 

 

 

 

 

 

IHDF CLVSKVVGRC RASMPRWWYNVTDGSCQLFV YGGCDGNSNN50 YLTKEECLKK CATV;

64 (SEQ ID NO;

6)YEEYCTANA VTGPCRASFP RWYFDVERNSCNNFIYGGCR GNKNSYRSEE150 ACMLRCFRQ;

159 (SEQ ID NO;

3) 

 

 

 

 

 

IHDF CLVSKVVGRC RASMPRWWYNVTDGSCQLFV YGGCDGNSNN50 YLTKEECLKK CATVTENATG DLATSRNAADSSVPSAPRRQ DSEDHSSDMF100 NYEEYCTANA VTGPCRASFP RWYFDVERNSCNNFIYGGCR GNKNSYRSEE150 ACMLRCFRQ;

159 (SEQ ID NO;

50) 

 

 

 

 

 

 

 

 

 

 

CLVSKVVGRC RASMPRWWYNVTDGSCQLFV YGGCDGNSNN50 YLTKEECLKK CATVTENATG DLATSRNAADSSVPSAPRRQ DSEDHSSDMF100 NYEEYCTANA VTGPGRASFP RWYFDVERNSCNNFIYGGCR GNKNSYRSEE150 ACMLRC; and

156 (SEQ ID NO;

1)ADRERSIHDF CLVSKVVGRC RASMPRWWYNVTDGSCQLFV YGGCDGNSNN50 YLTKEECLKK CATVTENATG DLATSRNAADSSVPSAPRRQ DSEDHSSDMF100 NYEEYCTANA VTGPCRASFP RWYFDVERNSCNNFIYGGCR GNKNSYRSEE150 ACMLRCFRQQ ENPPLPLGSK VVVLAGAVS.179

2. The nucleic acid sequence of claim 1, wherein the nucleic acid comprises a sequence selected from the group consisting of SEQ ID NOS:

9, 32, 44, 46, 48 and 51.

3. A self-replicating expression vector, comprising a nucleic acid sequence of claim 1 or 2.

4. The expression vector of claim 3, wherein the expression vector expresses said protein in an appropriate protein expression system that:

(a) is glycosylated;

(b) contains at least one intra-chain cysteine-cysteine disulfide bond;

or

(c) is glycosylated and contains at least one intra-chain cysteine-cysteine disulfide bond.

5. A method of producing a protein encoded by a nucleic acid of claim 1, comprising:

(a) inserting the nucleic acid into an appropriate protein expression vector by use of a recombinant DNA technology, to create a bikunin expression vector; and

(b) subjecting the bikunin expression vector to an appropriate protein expression system.

6. The method of claim 5, wherein the protein:

(a) is glycosylated;

(b) contains at least one intra-chain cysteine-cysteine disulfide bond;

or

(c) is both glycosylated and contains at least one intra-chain cysteine-cysteine disulfide bond.