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US6020313A
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Method for inducing bone formation using an extract of human osteosarcoma cell line SAOS-2
Assignee: KANSAS C/O OFFICE OF RESEARCH AND GRADUATE STUDIES, University of
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Filing: 1993-02-08
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Granted: 2000-02-01
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Expires: 2013-02-08
Abstract
Abstract The invention relates to a method for inducing bone formation via administration of an extract of devitalized, freeze dried Saos-2 cells. The extract is prepared by contacting the freeze dried Saos-2 cells, which are devitalized in the act of freeze drying, with a weak denaturing agent of the type used to separate proteins from cells. The extract does have properties superior to comparable treatment using whole Saos-2 cells. The extract is then further treated by applying it to a removing separating gel, and then fractions from the gel which have a molecular weight of from about 10 kd to about 60 kd. Also described are the extract itself, and formulations containing it.
Inventors
- 👤 H. Clarke Anderson
- 👤 Howard H. T. Hsu
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 09:41
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/475; C07K14/51; A; A61; A61K; A61K35/00; A61K35/12; A61K35/13; Y; Y10; Y10S; Y10S530/00; Y10S530/827; Y10S530/828
Claims Analysis
Total Claims: 25
Claim 1
Independent Claim
122 words
Method for inducing bone formation in a mammal comprising administering an amount of an extract of Saos-2 cells sufficient to promote bone healing or bone formation to a mammal in need of said bone healing or bone formation, said extract of Saos-2 cells obtained by contacting a sample of devitalized, freeze dried Saos-2 cells with a denaturing agent in an amount and concentration to permit refolding of any denatured protein to active form or insufficient to denature protein to form a supernatant and a precipitate, applying said supernatant to a protein absorbing gel, and removing protein fractions from said absorbing gel having a molecular weight range of from about 10 kd to about 60 kd, to form an extract of Saos-2 cells.
Claim 2
Dependent Claim
122 words
Method for inducing bone formation in a mammal comprising administering an amount of an extract of Saos-2 cells sufficient to promote bone healing or bone formation to a mammal in need of said bone healing or bone formation, said extract of Saos-2 cells obtained by contacting a sample of devitalized, freeze dried Saos-2 cells with a denaturing agent in an amount and concentration to permit refolding of any denatured protein to active form or insufficient to denature protein to form a supernatant and a precipitate, applying said supernatant to a protein absorbing gel, and removing protein fractions from said absorbing gel having a molecular weight range of from about 10 kd to about 60 kd, to form an extract of Saos-2 cells.
Claim 3
Dependent Claim
26 words
Method of claim 1, wherein said denaturing agent is a solution of guanidium hydrochloride at a concentration which permits a protein to refold to active form.
Claim 4
Dependent Claim
21 words
Method of claim 1, wherein said denaturing agent is a solution of urea at a concentration insufficient to denature a protein.
Claim 5
Dependent Claim
15 words
Method of claim 2, wherein said guanidium hydrochloride is at a concentration of about 4M.
Claim 6
Dependent Claim
14 words
Method of claim 3, wherein said urea is at a concentration of about 6M.
Claim 7
Dependent Claim
11 words
Method of claim 1, wherein said devitalized Saos-2 cells are defatted.
Claim 8
Dependent Claim
30 words
Method of claim 1, wherein said mammal is in need of healing of a bone defect treatment of a bone tumor or of promotion of assimilation of a bone graft.
Claim 9
Dependent Claim
15 words
Method of claim 1, further comprising lyophilizing said extract prior to administering to said mammal.
Claim 10
Dependent Claim
16 words
Method of claim 1, comprising combining said extract with collagen prior to administering to said mammal.
Claim 11
Dependent Claim
116 words
Method for inducing bone formation in a mammal comprising administering an amount of an extract of Saos-2 cells sufficient to promote bone healing or bone formation to a mammal in need of said bone healing or bone formation, said extract of Saos-2 cells obtained by contacting a sample of devitalized, freeze dried Saos-2 cells with a denaturing agent in an amount and concentration to permit refolding of any denatured protein to active form or insufficient to denature protein to form a supernatant and a precipitate, removing said supernatant from said precipitate, resuspending said precipitate in a solution, treating said solution to form a second supernatant and a second precipitate, wherein said second precipitate is said extract.
Claim 12
Dependent Claim
116 words
Method for inducing bone formation in a mammal comprising administering an amount of an extract of Saos-2 cells sufficient to promote bone healing or bone formation to a mammal in need of said bone healing or bone formation, said extract of Saos-2 cells obtained by contacting a sample of devitalized, freeze dried Saos-2 cells with a denaturing agent in an amount and concentration to permit refolding of any denatured protein to active form or insufficient to denature protein to form a supernatant and a precipitate, removing said supernatant from said precipitate, resuspending said precipitate in a solution, treating said solution to form a second supernatant and a second precipitate, wherein said second precipitate is said extract.
Claim 13
Dependent Claim
89 words
Extract of Saos-2 cells useful in inducing bone formation, said extract prepared by contacting a sample of devitalized freeze dried Saos-2 cells with a solution of a denaturing agent in an amount and concentration to permit refolding of any denatured protein to active form or insufficient to denature protein to form a supernatant and a precipitate, separating said supernatant therefrom, applying said supernatant to a protein absorbing gel and removing fractions from said protein absorbing gel having a molecular weight of from about 10 kd to about 60 kd.
Claim 14
Dependent Claim
89 words
Extract of Saos-2 cells useful in inducing bone formation, said extract prepared by contacting a sample of devitalized freeze dried Saos-2 cells with a solution of a denaturing agent in an amount and concentration to permit refolding of any denatured protein to active form or insufficient to denature protein to form a supernatant and a precipitate, separating said supernatant therefrom, applying said supernatant to a protein absorbing gel and removing fractions from said protein absorbing gel having a molecular weight of from about 10 kd to about 60 kd.
Claim 15
Dependent Claim
14 words
Extract of claim 11, wherein said denaturing agent is a solution of guanidium hydrochloride.
Claim 16
Dependent Claim
13 words
Extract of claim 11, wherein said denaturing agent is a solution of urea.
Claim 17
Dependent Claim
17 words
Extract of claim 12, wherein said solution of guanidium hydrochloride is at a concentration of about 4M.
Claim 18
Dependent Claim
16 words
Extract of claim 13, wherein said solution of urea is at a concentration of about 6M.
Claim 19
Dependent Claim
7 words
Extract of claim 11, in solution form.
Claim 20
Dependent Claim
7 words
Extract of claim 11, in pellet form.
Claim 21
Dependent Claim
7 words
Extract of claim 11, in lyophilized form.
Claim 22
Dependent Claim
17 words
Composition useful in inducing bone formation comprising the extract of claim 11 and a pharmaceutically acceptable carrier.
Claim 23
Dependent Claim
10 words
Composition of claim 19, wherein said pharmaceutical carrier is biodegradable.
Claim 24
Dependent Claim
12 words
Composition of claim 20, wherein said biodegradable pharmaceutical carrier is a matrix.
Claim 25
Dependent Claim
14 words
Composition useful in inducing bone formation comprising the extract of claim 11 and collagen.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2003-01-31 | 2003-08-02 | 2004-02-01 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2007-01-31 | 2007-08-02 | 2008-02-01 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2011-01-31 | 2011-08-02 | 2012-02-01 | $7700.00 | Window Closed / Lapsed |