US4698301A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Angiogenic factor production
Assignee: University of Manchester
•
Filing: 1983-11-29
•
Granted: 1987-10-06
•
Expires: 2003-11-29
Abstract
Abstract New angiogenic factors of low molecular weight (under 1000), useful as treatment agents for wound healing and for biological assaying, made from in vitro cultures of a wide variety of tumor cells by a treatment which breaks up the proteinaceous tumor angiogenesis factors ("TAF") in them and liberates the more mobile low molecular weight factors. The treatment can be applied to cell extracts or the aqueous portion of the culture, and the proteinaceous part may be rendered insoluble or captured chemically so as to release the new factor. Separation techniques which can be used include solvent extraction of dried liquid fractions, using polar solvents such as ethanol, and the liberated factor can be purified chromatographically using adsorbents for which the factor has affinity, especially a basic adsorbent and preferably polysaccharide-based, followed by elution.
Inventors
- 👤 Jacqueline B. Weiss
- 👤 Christopher R. Hill
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 05:36
CPC Classifications
A; A61; A61K; A61K35/00; A61K35/12; A61K35/13
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
112 words
A method for the preparation of an angiogenic factor of molecular weight of between 300 and 600, and having the ability to activate pro-collagenase, which comprises the steps of: (1) growing tumour cells in vitro in an aqueous nutrient culture medium by a suspension culture procedure so that said factor is released into the culture medium, (2) separating the grown cells from the culture medium to obtain an extract containing said angiogenic factor in admixture with proteinaceous materials, (3) treating the extract from step (2) to remove proteinaceous materials from it, and (4) recovering the angiogenic factor of molecular weight of between 300 and 600 from the extract remaining from step (3).
Claim 2
Independent Claim
112 words
A method for the preparation of an angiogenic factor of molecular weight of between 300 and 600, and having the ability to activate pro-collagenase, which comprises the steps of: (1) growing tumour cells in vitro in an aqueous nutrient culture medium by a suspension culture procedure so that said factor is released into the culture medium, (2) separating the grown cells from the culture medium to obtain an extract containing said angiogenic factor in admixture with proteinaceous materials, (3) treating the extract from step (2) to remove proteinaceous materials from it, and (4) recovering the angiogenic factor of molecular weight of between 300 and 600 from the extract remaining from step (3).
Claim 3
Dependent Claim
45 words
A method as claimed in claim 1 wherein the separated culture medium from step (2) is evaporated to dryness and the dried residue is extracted with a sufficient amount of a solvent which dissolves the low molecular weight angiogenic factor but not the proteinaceous material.
Claim 4
Dependent Claim
42 words
A method as claimed in claim 1 wherein the separated culture medium from step (2) is treated to render the proteinaceous material insoluble, and the insolubilised proteinaceous material is separated to leave an aqueous solution containing the low molecular weight angiogenic factor.
Claim 5
Dependent Claim
65 words
A method as claimed in claim 1 wherein the separated culture medium from step (2) is treated with a material which competes with the low molecular weight angiogenic factor for the proteinaceous materials and thereby releases the low molecular weight angiogenic factor from combination with the proteinaceous material, and then separating an aqueous fraction containing the low molecular weight angiogenic factor by dialysis and/or diafiltration.
Claim 6
Dependent Claim
33 words
A method as claimed in claim 1 wherein the low molecular weight angiogenic factor recovered by separation from the proteinaceous material is purified by adsorption on a basic adsorbent, and then eluted therefrom.
Claim 7
Dependent Claim
40 words
A method as claimed in claim 1 wherein the low molecular weight angiogenic factor recovered by separation from the proteinaceous material is purified by adsorption on an adsorbent comprising a fatty material on an inorganic substrate, and then eluted therefrom.
Claim 8
Dependent Claim
35 words
A method as claimed in claim 1 wherein the low molecular weight angiogenic factor recovered by separation from the proteinaceous material is purified by adsorption on an adsorbent comprising a collagen and then eluted therefrom.
Claim 9
Dependent Claim
51 words
A low molecular weight angiogenic factor which is anionic and water soluble, has the ability to activate procollagenase, has a molecular weight of between 300 and 600, binds to C 18 -coated silica, is CAM-active, and stimulates mitosis in capillary endothelial cells, obtained by a process as claimed in claim 1.
Claim 10
Dependent Claim
18 words
A composition comprising a low molecular weight angiogenic factor as claimed in claim 8 in combination with heparin.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1990-10-05 | 1991-04-06 | 1991-10-06 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1994-10-05 | 1995-04-06 | 1995-10-06 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1998-10-05 | 1999-04-06 | 1999-10-06 | $7700.00 | Window Closed / Lapsed |