US5057540A
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Saponin adjuvant
Assignee: Cambridge Biotech Corp
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Filing: 1990-08-27
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Granted: 1991-10-15
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Expires: 2010-08-27
Abstract
Abstract Substantially pure saponins are disclosed. The saponins of the present invention are useful as immune adjuvants. Disclosed as well are immune response-provoking compositions comprising an antigen in admixture with the substantially pure saponins.
Inventors
- 👤 Charlotte A. Kensil
- 👤 Dante J. Marciani
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 20:43
CPC Classifications
C; C07; C07J; C07J17/00; C07J17/005; A; A61; A61K; A61K39/00; A61K39/39; C07H; C07H15/00; C07H15/20; C07H15/24; C07H15/256; A61K2039/555; A61K2039/55511; A61K2039/55577
Claims Analysis
Total Claims: 21
Claim 1
Independent Claim
105 words
Substantially pure saponin purified from a crude Quillaja saponaria extract wherein said pure saponin is characterized by a single predominant peak which comprises 90% or more of the total area of all peaks of a chromatogram, excluding the solvent peak, when analyzed on reverse phase-HPLC on a Vydac C 4 column having 5 μm particle size, 330 Å pore, 4.6 mm ID×25 cm L in a solvent of 40 mM acetic acid in methanol/water (58/42; v/v) at a flow rate of 1 ml/mixture, and wherein said saponin has immune adjuvant activity and is less toxic when used as an adjuvant than said Quillaja saponaria extract.
Claim 2
Dependent Claim
105 words
Substantially pure saponin purified from a crude Quillaja saponaria extract wherein said pure saponin is characterized by a single predominant peak which comprises 90% or more of the total area of all peaks of a chromatogram, excluding the solvent peak, when analyzed on reverse phase-HPLC on a Vydac C 4 column having 5 μm particle size, 330 Å pore, 4.6 mm ID×25 cm L in a solvent of 40 mM acetic acid in methanol/water (58/42; v/v) at a flow rate of 1 ml/mixture, and wherein said saponin has immune adjuvant activity and is less toxic when used as an adjuvant than said Quillaja saponaria extract.
Claim 3
Dependent Claim
93 words
Substantially pure QA-7 saponin purified from a crude Quillaja saponaria extract wherein said pure saponin is characterized by one predominant peak which comprises 90% or more of the total area of all peaks of a chromatogram, excluding the solvent peak, and having a retention time of approximately 9-10 minutes when analyzed on reverse phase HPLC on a Vydac C 4 column having 5 μm particle size, 330 Å pore, 4.6 mm ID×25 cm L in a solvent of 40 mM acetic acid in methanol/water (58/42; v/v) at a flow rate of 1 ml/minute.
Claim 4
Dependent Claim
93 words
Substantially pure QA-7 saponin purified from a crude Quillaja saponaria extract wherein said pure saponin is characterized by one predominant peak which comprises 90% or more of the total area of all peaks of a chromatogram, excluding the solvent peak, and having a retention time of approximately 9-10 minutes when analyzed on reverse phase HPLC on a Vydac C 4 column having 5 μm particle size, 330 Å pore, 4.6 mm ID×25 cm L in a solvent of 40 mM acetic acid in methanol/water (58/42; v/v) at a flow rate of 1 ml/minute.
Claim 5
Dependent Claim
73 words
The substantially pure QA-7 saponin of claim 2, wherein said saponin has immune adjuvant activity, and wherein said saponin is characterized by a carbohydrate content of about 35% per dry weight as assayed by anthrone, has a UV adsorption maxima of 205-210 nm, has a micellar concentration of 0.06% (w/v) in water and 0.07% in phosphate buffered saline, and causes no detectable hemolysis of sheep red blood cells at concentrations of 200 μg/ml.
Claim 6
Dependent Claim
33 words
The substantially pure QA-7 saponin of claim 3, wherein said carbohydrate content has a composition comprising the monosuccharides: terminal rhamnose, terminal xylose, terminal glucose, terminal galactose, 3-xylose, 3,4-rhamnose, 2,3-fucose, 2,3-glucuronic acid and apiose.
Claim 7
Dependent Claim
91 words
Substantially pure QA-21 saponin purified from a crude Quillaja saponaria extract wherein said pure saponin is characterized by one predominant peak which comprises 90% or more of the total area of all peaks of a chromatogram, excluding the solvent peak, and having a retention time of approximately 51minutes when analyzed on reverse phase-HPLC on a Vydac C 4 column having 5 μm particle size, 330 Å pore, 4.6 mm ID×25 cm L in a solvent of 40 mM acetic acid in methanol/water (58/42; v/v) at a flow rate of 1 ml/minute.
Claim 8
Dependent Claim
91 words
Substantially pure QA-21 saponin purified from a crude Quillaja saponaria extract wherein said pure saponin is characterized by one predominant peak which comprises 90% or more of the total area of all peaks of a chromatogram, excluding the solvent peak, and having a retention time of approximately 51minutes when analyzed on reverse phase-HPLC on a Vydac C 4 column having 5 μm particle size, 330 Å pore, 4.6 mm ID×25 cm L in a solvent of 40 mM acetic acid in methanol/water (58/42; v/v) at a flow rate of 1 ml/minute.
Claim 9
Dependent Claim
75 words
The substantially pure QA-21 saponin of claim 5, wherein said saponin has immune adjuvant activity, and wherein said saponin is characterized by a carbohydrate content of about 22% per dry weight as assayed by anthrone, has a UV absorption maxima of 205-210 nm, has a micellar concentration of about 0.03% (w/v) in water and 0.02% (w/v) in phosphate buffered saline, and causes hemolysis of sheep red blood cells at concentrations of 25 μg/ml or greater.
Claim 10
Dependent Claim
37 words
The substantially pure QA-21 saponin of claim 6, wherein said carbohydrate content has a composition comprising the monosaccharides: terminal rhamnose, terminal arabinose, terminal apiose, terminal xylose, 4-rhamnose, terminal glucose, terminal galactose, 2-fucose, 3-xylose, 3,4-rhamnose and 2,3-glucuronic acid.
Claim 11
Dependent Claim
91 words
A substantially pure QA-17 saponin purified from a crude Quillaja saponaria extract wherein said pure saponin is characterized by one predominant peak which comprises 90% or more of the total area of all peaks of a chromatogram, excluding the solvent peak, and having a retention time of approximately 35 minutes on reverse phase-HPLC on a Vydac C 4 column having 5 μm particle size, 330 Å pore, 4.6 mm ID×25 cm L in a solvent of 40 mN acetic acid in methanol/water (58/42; v/v) at a flow rate of 1 ml/minute.
Claim 12
Dependent Claim
91 words
A substantially pure QA-17 saponin purified from a crude Quillaja saponaria extract wherein said pure saponin is characterized by one predominant peak which comprises 90% or more of the total area of all peaks of a chromatogram, excluding the solvent peak, and having a retention time of approximately 35 minutes on reverse phase-HPLC on a Vydac C 4 column having 5 μm particle size, 330 Å pore, 4.6 mm ID×25 cm L in a solvent of 40 mN acetic acid in methanol/water (58/42; v/v) at a flow rate of 1 ml/minute.
Claim 13
Dependent Claim
71 words
The substantially QA-17 saponin of claim 8, wherein said saponin has immune adjuvant activity, and wherein said saponin is characterized by a carbohydrate content of about 29% per dry weight as assayed by anthrone, has a UV absorption maxima of 205-210 nm, has a micellar concentration of about 0.06% (w/v) in water and 0.03% (w/v) in phosphate-bufferred saline, and causes hemolysis of sheep red blood cells at concentrations of 25 μg/ml.
Claim 14
Dependent Claim
34 words
The substantially pure QA-17 saponin of claim 9, wherein said carbohydrate content has a composition comprising the monosaccharides: terminal rhamnose, terminal xylose, 2-fucose, 3-xylose, 3,4-rhamnose, 2,3-glucuronic acid, terminal glucose, 2-arabinose, terminal galactose and apiose.
Claim 15
Dependent Claim
91 words
A substantially pure QA-18 saponin purified from a crude Quillaja saponaria extract wherein said pure saponin is characterized by one predominant peak which comprises 90% or more of the total area of all peaks of a chromatogram, excluding the solvent peak, and having a retention time of approximately 38 minutes on reverse phase-HPLC on a Vydac C 4 column having 5 μm particle size, 330 Å pore, 4.6 mm ID×25 cm L in a solvent of 40 mM acetic acid in methanol/water (58/42; v/v) at a flow rate of 1 ml/minute.
Claim 16
Dependent Claim
91 words
A substantially pure QA-18 saponin purified from a crude Quillaja saponaria extract wherein said pure saponin is characterized by one predominant peak which comprises 90% or more of the total area of all peaks of a chromatogram, excluding the solvent peak, and having a retention time of approximately 38 minutes on reverse phase-HPLC on a Vydac C 4 column having 5 μm particle size, 330 Å pore, 4.6 mm ID×25 cm L in a solvent of 40 mM acetic acid in methanol/water (58/42; v/v) at a flow rate of 1 ml/minute.
Claim 17
Dependent Claim
70 words
The substantially QA-18 saponin of claim 11, wherein said saponin has immune adjuvant activity, and wherein said saponin is characterized by a carbohydrate content of about 25-26% per dry weight as assayed by anthrone, has a UV absorption maxima of 205-210 nm, has a micellar concentration of 0.04% (w/v) in water and 0.02% (w/v) in phosphate-buffered saline, and causes hemolysis of sheep red blood cells at concentrations of 25 μg/ml.
Claim 18
Dependent Claim
36 words
The substantially pure QA-18 saponin of claim 12, wherein said carbohydrate content has a composition comprising the monosaccharides: terminal rhamnose, terminal arabinose, terminal apiose, terminal xylose, terminal glucose, terminal galatose, 2-fucose, 3-xylose, 3,4-rhamnose and 2,3-glucuronic acid.
Claim 19
Independent Claim
49 words
A method of enhancing an immune response to an antigen in an individual comprising administration of an amount of the substantially pure saponin adjuvants from any of claims 1-7 and 8-13 to said individual in an amount sufficient to enhance the immune response of said individual to said antigen.
Claim 20
Dependent Claim
62 words
A pharmaceutical composition useful for inducing the production of antibodies to an antigen in an individual comprising an immunogenically effective amount of an antigen and at least one substantially pure saponin as in any one of claims 1-7 and 8-13, wherein said substantially pure saponin is present in an amount sufficient to enhance the immune response of said individual to said antigen.
Claim 21
Dependent Claim
12 words
The pharmaceutical composition of claim 15, wherein said individual is a mammal.
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| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1994-10-14 | 1995-04-15 | 1995-10-15 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-10-14 | 1999-04-15 | 1999-10-15 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-10-14 | 2003-04-15 | 2003-10-15 | $7700.00 | Window Closed / Lapsed |