US5250414A
Expired / Lapsed
US
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Diagnostic methods using neurite growth regulatory factors
Assignee: Zurich Universität
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Filing: 1991-06-24
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Granted: 1993-10-05
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Expires: 2011-06-24
Abstract
Abstract The proteins of the present invention include central nervous system myelin associated proteins and metalloproteases associated with glioblastoma cells and other malignant tumors which can metastasize to the brain. The CNS myelin associated proteins inhibit neurite outgrowth in nerve cells and neuroblastoma cells, and can also inhibit fibroblast spreading. Such inhibitory proteins include a 35,000 dalton and a 250,000 dalton molecular weight protein. The CNS myelin associated inhibitory proteins may be used in the treatment of malignant tumors. Antibodies to the CNS myelin associated proteins can be used in the diagnosis and therapies of nerve damage. Monoclonal antibody IN-1 may be used to promote regeneration of nerve fibers over long distances in spinal cord lesions. The metalloproteases of the invention have value in diagnosis of malignancies and the treatment of nerve damage and degenerative disorders of the nervous system. Inhibitors of the metalloproteases in combination with the CNS myelin associated inhibitory proteins can be used in the treatment of malignant tumors. Methods of determining malignant potential of a cell by measuring metalloprotease activity are provided.
Inventors
- 👤 Martin E. Schwab
- 👤 Pierenrico W. Caroni
- 👤 Paolo A. Paganetti
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-28 10:39
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/64; C12N9/6421; C12N9/6489; C07; C07K; C07K14/00; C07K14/435; C07K14/475; C07K14/705; C07K14/71; C07K16/00; C07K16/18; C07K16/22; C07K16/40; C07K5/00; C07K5/04; C07K5/10; C07K5/1002; C07K5/1016; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/575; G01N33/5758; G01N33/74; A; A61; A61K; A61K38/00; Y; Y10; Y10S; Y10S436/00; Y10S436/811; Y10S436/813
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
144 words
A method for diagnosing a malignancy invasive in central nervous system white matter in a patient in vitro comprising detecting the presence in a sample of a plasma membrane-associated metalloprotease characterized by the following properties: (a) a metalloendoprotease activity which cleaves under suitable conditions the peptide benzyloxycarbonyl-Phe-Ala-Phe-Tyr, which activity is maximally active at a pH in the range of 5.5 to 7.0 and is substantially lost in 1% (w/v) CHAPS; and (b) the ability to neutralize the inhibitory substrate property of central nervous system myelin of a higher vertebrate, in which the neutralization is detected by observing the ability of the central nervous system myelin in the presence of the metalloprotease to support neurite outgrowth or fibroblast spreading in vitro; said sample containing cells of the patient suspected of being malignant; in which the presence of the metalloprotease indicates the presence of the malignancy.
Claim 2
Independent Claim
144 words
A method for diagnosing a malignancy invasive in central nervous system white matter in a patient in vitro comprising detecting the presence in a sample of a plasma membrane-associated metalloprotease characterized by the following properties: (a) a metalloendoprotease activity which cleaves under suitable conditions the peptide benzyloxycarbonyl-Phe-Ala-Phe-Tyr, which activity is maximally active at a pH in the range of 5.5 to 7.0 and is substantially lost in 1% (w/v) CHAPS; and (b) the ability to neutralize the inhibitory substrate property of central nervous system myelin of a higher vertebrate, in which the neutralization is detected by observing the ability of the central nervous system myelin in the presence of the metalloprotease to support neurite outgrowth or fibroblast spreading in vitro; said sample containing cells of the patient suspected of being malignant; in which the presence of the metalloprotease indicates the presence of the malignancy.
Claim 3
Dependent Claim
13 words
The method according to claim 1 in which the malignancy is a glioblastoma.
Claim 4
Independent Claim
107 words
A method of determining the malignant potential of a cell suspected of being invasive in the central nervous system comprising measuring in vitro a plasma membrane-associated metallodependent degradative activity of the cell; in which the metallodependent degradative activity neutralizes under suitable conditions the inhibitory substrate property of central nervous system myelin of a higher vertebrate, in which the neutralization is detected by observing the ability of the central nervous system myelin in the presence of the metallodependent degradative activity to support neurite outgrowth or fibroblast spreading in vitro; and in which the amount of metallodependent degradative activity of the cell indicates the malignant potential of the cell.
Claim 5
Independent Claim
107 words
A method of determining the malignant potential of a cell suspected of being invasive in the central nervous system comprising measuring in vitro a plasma membrane-associated metallodependent degradative activity of the cell; in which the metallodependent degradative activity neutralizes under suitable conditions the inhibitory substrate property of central nervous system myelin of a higher vertebrate, in which the neutralization is detected by observing the ability of the central nervous system myelin in the presence of the metallodependent degradative activity to support neurite outgrowth or fibroblast spreading in vitro; and in which the amount of metallodependent degradative activity of the cell indicates the malignant potential of the cell.
Claim 6
Dependent Claim
21 words
The method according to claim 3 in which the metallodependent degradative activity is measured by an in vitro peptide substrate assay.
Claim 7
Dependent Claim
38 words
The method according to claim 4 in which the assay is carried out by determining the concentration of peptide substrate necessary to achieve a defined level of inhibition of spreading by the cell on central nervous system myelin.
Claim 8
Dependent Claim
13 words
The method according to claim 5 in which the peptide substrate is benzyloxycarbonyl-Phe-Ala-Phe-Tyr.
Claim 9
Independent Claim
81 words
A method of determining the malignant potential of a cell suspected of being invasive in the central nervous system comprising measuring in vitro a plasma membrane-associated metalloendoproteolytic activity of the cell, which activity: (a) cleaves under suitable conditions the peptide benzyloxycarbonyl-Phe-Ala-Phe-Tyr; (b) is maximally active at a pH in the range of 5.5 to 7.0; and (c) is substantially lost in 1% (w/v) CHAPS; in which the amount of metalloendoproteolytic activity of the cell indicates the malignant potential of the cell.
Claim 10
Independent Claim
81 words
A method of determining the malignant potential of a cell suspected of being invasive in the central nervous system comprising measuring in vitro a plasma membrane-associated metalloendoproteolytic activity of the cell, which activity: (a) cleaves under suitable conditions the peptide benzyloxycarbonyl-Phe-Ala-Phe-Tyr; (b) is maximally active at a pH in the range of 5.5 to 7.0; and (c) is substantially lost in 1% (w/v) CHAPS; in which the amount of metalloendoproteolytic activity of the cell indicates the malignant potential of the cell.
Claim 11
Dependent Claim
20 words
The method according to claim 7 in which the metalloendoproteolytic activity is measured by an in vitro peptide substrate assay.
Claim 12
Dependent Claim
38 words
The method according to claim 8 in which the assay is carried out by determining the concentration of peptide substrate necessary to achieve a defined level of inhibition of spreading by the cell on central nervous system myelin.
Claim 13
Dependent Claim
13 words
The method according to claim 9 in which the peptide substrate is benzyloxycarbonyl-Phe-Ala-Phe-Tyr.
Claim 14
Independent Claim
61 words
A method of determining the malignant potential of a cell suspected of being invasive in the central nervous system comprising measuring in vitro the amount of a metalloprotease inhibitor necessary to achieve a defined level of inhibition of spreading by the cell on central nervous system myelin, in which the amount of metalloprotease inhibitor indicates the malignant potential of the cell.
Claim 15
Independent Claim
61 words
A method of determining the malignant potential of a cell suspected of being invasive in the central nervous system comprising measuring in vitro the amount of a metalloprotease inhibitor necessary to achieve a defined level of inhibition of spreading by the cell on central nervous system myelin, in which the amount of metalloprotease inhibitor indicates the malignant potential of the cell.
Claim 16
Dependent Claim
18 words
The method according to claim 11 in which the metalloprotease inhibitor is a peptide substrate of the metalloprotease.
Claim 17
Independent Claim
58 words
A method of determining the malignant potential of a cell suspected of being invasive in the central nervous system comprising measuring in vitro the amount of benzyloxycarbonyl-Phe-Ala-Phe-Tyr necessary to achieve a defined level of inhibition of spreading by the cell on central nervous system myelin, in which the amount of benzyloxycarbonyl-Phe-Ala-Phe-Tyr indicates the malignant potential of the cell.
Claim 18
Independent Claim
58 words
A method of determining the malignant potential of a cell suspected of being invasive in the central nervous system comprising measuring in vitro the amount of benzyloxycarbonyl-Phe-Ala-Phe-Tyr necessary to achieve a defined level of inhibition of spreading by the cell on central nervous system myelin, in which the amount of benzyloxycarbonyl-Phe-Ala-Phe-Tyr indicates the malignant potential of the cell.
Claim 19
Dependent Claim
17 words
The method according to claim 3, 7 or 4 in which the cell is a glial cell.
Claim 20
Dependent Claim
17 words
The method according to claim 8, 5 or 9 in which the cell is a glial cell.
Claim 21
Dependent Claim
17 words
The method according to claim 6, 10 or 11 in which the cell is a glial cell.
Claim 22
Dependent Claim
16 words
The method according to claim 12 or 13 in which the cell is a glial cell.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1996-10-04 | 1997-04-05 | 1997-10-05 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2000-10-04 | 2001-04-05 | 2001-10-05 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2004-10-04 | 2005-04-05 | 2005-10-05 | $7700.00 | Window Closed / Lapsed |