US4996159A
Expired / Lapsed
US
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Neovascularization inhibitors and methods for their production and use
Assignee: Johns Hopkins University
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Filing: 1990-01-26
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Granted: 1991-02-26
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Expires: 2010-01-26
Abstract
Abstract Neovascularization inhibitors are disclosed, which are purified polypeptides recovered from cultured cells, including retinal pigment epithelial cells and human fibroblast cells. The polypeptides may be used for the treatment of diseases in which new blood vessel formation plays a role, such as diabetic retinopathy, senile macular degeneration, tumor growth, and rheumatoid arthritis.
Inventors
- 👤 Bert M. Glaser
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-10 01:48
CPC Classifications
C; C07; C07K; C07K16/00; C07K16/18; A; A61; A61P; A61P9/00; C07K14/00; C07K14/435; C07K14/46; C07K14/47; C07K14/4701; C07K14/4702; C07K14/4703; A61K; A61K38/00; Y; Y10; Y10S; Y10S530/00; Y10S530/827; Y10S530/849
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
66 words
A process for producing a neovascularization inhibitor comprising the steps of: culturing retinal pigment epithelial cells to produce a culture medium containing said neovascularization inhibitor; and purifying said neovascularization inhibitor from said medium wherein said neovascularization inhibitor has a molecular weight of approximately 57,000±3,000 and an isoelectric point of approximately 4.6±0.3 and is stable in an environment with a pH of approximately from 2 to 3.
Claim 2
Dependent Claim
66 words
A process for producing a neovascularization inhibitor comprising the steps of: culturing retinal pigment epithelial cells to produce a culture medium containing said neovascularization inhibitor; and purifying said neovascularization inhibitor from said medium wherein said neovascularization inhibitor has a molecular weight of approximately 57,000±3,000 and an isoelectric point of approximately 4.6±0.3 and is stable in an environment with a pH of approximately from 2 to 3.
Claim 3
Dependent Claim
22 words
A process as recited in claim 1 wherein said retinal pigment epithelial cells are obtained from animals eyes that lack a tapetum.
Claim 4
Dependent Claim
14 words
A process as recited in claim 2 wherein said animal eyes are human eyes.
Claim 5
Dependent Claim
14 words
A process as recited in claim 2 wherein said animal eyes are pig eyes.
Claim 6
Dependent Claim
77 words
A process for producing a neovascularization inhibitor comprising the steps of: culturing retinal pigment epithelial cells to produce a culture medium containing said neovascularization inhibitor; separating said medium from said cells; and substantially purifying said inhibitor by subjecting said medium to hydrophobic interaction chromatography wherein said neovascularization inhibitor has a molecular weight of approximately 57,000±3,000 and an isoelectric point of approximately 4.6±0.3 and is stable in an environment with a pH of approximately from 2 to 3.
Claim 7
Dependent Claim
77 words
A process for producing a neovascularization inhibitor comprising the steps of: culturing retinal pigment epithelial cells to produce a culture medium containing said neovascularization inhibitor; separating said medium from said cells; and substantially purifying said inhibitor by subjecting said medium to hydrophobic interaction chromatography wherein said neovascularization inhibitor has a molecular weight of approximately 57,000±3,000 and an isoelectric point of approximately 4.6±0.3 and is stable in an environment with a pH of approximately from 2 to 3.
Claim 8
Dependent Claim
27 words
A process as recited in claim 5 comprising the additional step of purifying said substantially purified inhibitor by subjecting said substantially purified inhibitor to size exclusion chromatography.
Claim 9
Dependent Claim
23 words
A process as recited in claim 6 wherein said dialysis is employed at any stage after said medium is separated from said cells.
Claim 10
Dependent Claim
34 words
A process as recited in claim 7 wherein said acid dialysis is employed after said medium is separated from said cells and before said step of substantially purifying said inhibitor by hydrophobic interaction chromatography.
Claim 11
Dependent Claim
105 words
A process for obtaining a neovascularization inhibitor produced by retinal pigment epithelial cells comprising the steps of: culturing retinal pigment epithelial cells to produce a culture medium containing a neovascularization inhibitor wherein said neovascularization inhibitor has a molecular weight of approximately 57,000±3,000 and an isoelectric point of approximately 4.6±0.3 and is stable in an environment with a pH of approximately from 2 to 3; separating said medium from said cells; bringing said medium into contact with immobilized antibodies to said neovascularization inhibitor so as to bind said neovascularization inhibitor in said medium to said immobilized antibodies; and separating said neovascularization inhibitor from said immobilized antibodies.
Claim 12
Dependent Claim
105 words
A process for obtaining a neovascularization inhibitor produced by retinal pigment epithelial cells comprising the steps of: culturing retinal pigment epithelial cells to produce a culture medium containing a neovascularization inhibitor wherein said neovascularization inhibitor has a molecular weight of approximately 57,000±3,000 and an isoelectric point of approximately 4.6±0.3 and is stable in an environment with a pH of approximately from 2 to 3; separating said medium from said cells; bringing said medium into contact with immobilized antibodies to said neovascularization inhibitor so as to bind said neovascularization inhibitor in said medium to said immobilized antibodies; and separating said neovascularization inhibitor from said immobilized antibodies.
Claim 13
Dependent Claim
100 words
A process for producing a neovascularization inhibitor comprising the steps of: growing human retinal pigment epithelial cells in a culture medium; separating said medium from said cells; subjecting said separated medium to acid dialysis to produce a dialysate containing said inhibitor; substantially purifying said inhibitor in said dialysate by reverse-phase high performance liquid chromatography; and purifying said substantially purified inhibitor by gel permeation high performance liquid chromatography wherein said neovascularization inhibitor has a molecular weight of approximately 57,000±3,000 and an isoelectric point of approximately 4.6±0.3 ad is stable in an environment with a pH of approximately from 2 to 3.
Claim 14
Dependent Claim
100 words
A process for producing a neovascularization inhibitor comprising the steps of: growing human retinal pigment epithelial cells in a culture medium; separating said medium from said cells; subjecting said separated medium to acid dialysis to produce a dialysate containing said inhibitor; substantially purifying said inhibitor in said dialysate by reverse-phase high performance liquid chromatography; and purifying said substantially purified inhibitor by gel permeation high performance liquid chromatography wherein said neovascularization inhibitor has a molecular weight of approximately 57,000±3,000 and an isoelectric point of approximately 4.6±0.3 ad is stable in an environment with a pH of approximately from 2 to 3.
Claim 15
Dependent Claim
35 words
The process of claim 9 wherein said step of separating comprises lowering the pH of the environment of the inhibitor-antibody complex to approximately from 2 to 3 and eluting said inhibitor from said immobilized antibodies.
Claim 16
Dependent Claim
11 words
The process of claim 9 wherein said antibodies are polyclonal antibodies.
Claim 17
Dependent Claim
11 words
The process of claim 9 wherein said antibodies are monoclonal antibodies.
References Cited (Backward Citations)
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