US5266464A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Method of screening for protein inhibitors and activators
Assignee: Housey Pharmaceuticals Inc
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Filing: 1989-08-10
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Granted: 1993-11-30
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Expires: 2009-08-10
Abstract
Abstract Inhibitors and activators of a protein whose expression affects a phenotypic characteristic of the cell, especially a cultural or morphological characteristic, are identified by their more pronounced effect on cells producing higher, usually non-naturally occurring, levels of the protein, than on cells producing little or none of the protein. In a preferred assay, the effect is observable with the naked eye. By this method, tamoxifen is identified as an inhibitor of PKC activity in cell culture.
Inventors
- 👤 Gerard Housey
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 03:05
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/02; C12N; C12N9/00; C12N9/10; C12N9/12; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/5005; G01N33/5008
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
98 words
A test kit for determining whether a substance is an inhibitor or activator of a protein whose production evokes a responsive change in a phenotypic characteristic other than level of said protein in said cell per se, which comprises: (a) a first mammalian cell line which produces the protein and exhibits said phenotypic response thereto; and (b) a second mammalian cell line which produces the protein at a lower level than the first cell line, or does not produce the protein, and which exhibits said phenotypic response to the protein to a lesser degree or not at all.
Claim 2
Dependent Claim
98 words
A test kit for determining whether a substance is an inhibitor or activator of a protein whose production evokes a responsive change in a phenotypic characteristic other than level of said protein in said cell per se, which comprises: (a) a first mammalian cell line which produces the protein and exhibits said phenotypic response thereto; and (b) a second mammalian cell line which produces the protein at a lower level than the first cell line, or does not produce the protein, and which exhibits said phenotypic response to the protein to a lesser degree or not at all.
Claim 3
Dependent Claim
109 words
A test kit for determining whether a substance is an inhibitor or activator of a protein whose production evokes a responsive change in a phenotypic characteristic other than level of said protein in said cell per se, which comprises: (a) a first cell line which produces the protein and exhibits said phenotypic response thereto; (b) a second cell line which produces the protein at a lower level than the first cell line, or does not produce the protein, and which exhibits said phenotypic response to the protein to a lesser degree or not at all; and (c) a reference reagent comprising a known inhibitor or activator of the protein.
Claim 4
Dependent Claim
109 words
A test kit for determining whether a substance is an inhibitor or activator of a protein whose production evokes a responsive change in a phenotypic characteristic other than level of said protein in said cell per se, which comprises: (a) a first cell line which produces the protein and exhibits said phenotypic response thereto; (b) a second cell line which produces the protein at a lower level than the first cell line, or does not produce the protein, and which exhibits said phenotypic response to the protein to a lesser degree or not at all; and (c) a reference reagent comprising a known inhibitor or activator of the protein.
Claim 5
Independent Claim
144 words
A method of determining whether a substance is an inhibitor or activator of a protein whose production by a cell evokes a responsive change in a phenotypic characteristic other than the level of said protein in said cell per se, which comprises: (a) providing a first FDC-P1 cell line which produces the protein and exhibits said phenotypic response thereto; (b) providing a second FDC-P1 cell line which produces the protein at a lower level than the first cell line, or does not produce the protein at all, and which exhibits said phenotypic response to the protein to a lesser degree or not at all; (c) incubating the substance with the first and second FDC-P1 cell lines; and (d) comparing the phenotypic response of the first FDC-P1 cell line to the substance with the phenotypic response of the second FDC-P1 cell line to the substance.
Claim 6
Independent Claim
144 words
A method of determining whether a substance is an inhibitor or activator of a protein whose production by a cell evokes a responsive change in a phenotypic characteristic other than the level of said protein in said cell per se, which comprises: (a) providing a first FDC-P1 cell line which produces the protein and exhibits said phenotypic response thereto; (b) providing a second FDC-P1 cell line which produces the protein at a lower level than the first cell line, or does not produce the protein at all, and which exhibits said phenotypic response to the protein to a lesser degree or not at all; (c) incubating the substance with the first and second FDC-P1 cell lines; and (d) comparing the phenotypic response of the first FDC-P1 cell line to the substance with the phenotypic response of the second FDC-P1 cell line to the substance.
Claim 7
Dependent Claim
45 words
The method of claim 3 wherein said first cell line is obtained by introducing a gene encoding the protein of interest into a FDC-P1 host cell, said gene being under the control of a promoter functional in the FDC-P1 cell, whereby said gene is expressed.
Claim 8
Independent Claim
144 words
A method of determining whether a substance is an inhibitor or activator of a protein whose production by a cell evokes a responsive change in a phenotypic characteristic other than the level of said protein in said cell per se, which comprises: (a) providing a first CHO cell line which produces the protein and exhibits said phenotypic response thereto; (b) providing a second CHO cell line which produces the protein at a lower level than the first cell line, or does not produce the protein at all, and which exhibits said phenotypic response to the protein to a lesser degree or not at all; (c) incubating the substance with the first and second CHO cell lines; and (d) comparing the phenotypic response of the first CHO cell line to the substance with the phenotypic response of the second CHO cell line to the substance.
Claim 9
Independent Claim
144 words
A method of determining whether a substance is an inhibitor or activator of a protein whose production by a cell evokes a responsive change in a phenotypic characteristic other than the level of said protein in said cell per se, which comprises: (a) providing a first CHO cell line which produces the protein and exhibits said phenotypic response thereto; (b) providing a second CHO cell line which produces the protein at a lower level than the first cell line, or does not produce the protein at all, and which exhibits said phenotypic response to the protein to a lesser degree or not at all; (c) incubating the substance with the first and second CHO cell lines; and (d) comparing the phenotypic response of the first CHO cell line to the substance with the phenotypic response of the second CHO cell line to the substance.
Claim 10
Dependent Claim
45 words
The method of claim 5 wherein said first cell line is obtained by introducing a gene encoding the protein of interest into a CHO host cell, said gene being under the control of a promoter functional in the CHO cell, wherein said gene is expressed.
Claim 11
Independent Claim
119 words
A method of determining whether a substance is an inhibitor or activator of an insulin receptor which comprises: (a) providing a first cell line which produces an insulin receptor and exhibits a phenotypic response thereof; (b) providing a second cell line which produces the insulin receptor at a lower level than the first cell line, or does not produce the insulin receptor at all, and which exhibits a phenotypic response to the insulin receptor to a lesser degree or not at all; (c) incubating the substance with the first and second cell lines; and (d) comparing the phenotypic response of the first cell line to the substance with the phenotypic response of the second cell line to the substance.
Claim 12
Independent Claim
119 words
A method of determining whether a substance is an inhibitor or activator of an insulin receptor which comprises: (a) providing a first cell line which produces an insulin receptor and exhibits a phenotypic response thereof; (b) providing a second cell line which produces the insulin receptor at a lower level than the first cell line, or does not produce the insulin receptor at all, and which exhibits a phenotypic response to the insulin receptor to a lesser degree or not at all; (c) incubating the substance with the first and second cell lines; and (d) comparing the phenotypic response of the first cell line to the substance with the phenotypic response of the second cell line to the substance.
Claim 13
Independent Claim
118 words
A method of determining whether a substance is inhibitor or activator of a serotonin receptor, which comprises: (a) providing a first cell line which produces a serotonin receptor and exhibits a phenotypic response thereto; (b) providing a second cell line which produces the serotonin receptor at a lower level than the first cell line, or does not produce the serotonin receptor at all, and which exhibits a phenotypic response to the serotonin receptor to a lesser degree or not at all; (c) incubating the substance with the first and second cell lines; and (d) comparing the phenotypic response of the first cell line to the substance with the phenotypic response of the second cell line to the substance.
Claim 14
Independent Claim
118 words
A method of determining whether a substance is inhibitor or activator of a serotonin receptor, which comprises: (a) providing a first cell line which produces a serotonin receptor and exhibits a phenotypic response thereto; (b) providing a second cell line which produces the serotonin receptor at a lower level than the first cell line, or does not produce the serotonin receptor at all, and which exhibits a phenotypic response to the serotonin receptor to a lesser degree or not at all; (c) incubating the substance with the first and second cell lines; and (d) comparing the phenotypic response of the first cell line to the substance with the phenotypic response of the second cell line to the substance.
References Cited (Backward Citations)
US4480038A
Cited
US4532204A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1996-11-29 | 1997-05-31 | 1997-11-30 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2000-11-29 | 2001-05-31 | 2001-11-30 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2004-11-29 | 2005-05-31 | 2005-11-30 | $7700.00 | Window Closed / Lapsed |