Patent US4468379A successfully refreshed from Google Patents.
US4468379A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Leukocyte extracts for affecting the immune system
Assignee: Endeavor Corp
•
Filing: 1982-11-15
•
Granted: 1984-08-28
•
Expires: 2002-11-15
Abstract
Abstract A group of substances derived from human leukocytes (white blood cells) are described that amplify, suppress, or otherwise modulate the response of the immune system to reintroduction of antigens. The described materials are generically effective as to antigens rather than specific as to particular antigens, but the described modulators of immune response are effective only in respect of antigens with which the recipient has previously been challenged. The described materials are extracted from leukocyte dialysates by processes involving high pressure liquid chromatography with specified resin/solvent systems. Uses and pharmaceutical compositions for use of the materials are also described.
Inventors
- 👤 A. Arthur Gottlieb
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 05:56
CPC Classifications
A; A61; A61K; A61K45/00; A61K45/05; A61K35/00; A61K35/12; A61K35/14; A61K35/15; B; B01; B01D; B01D61/00; B01D61/24; B01D61/243; B01D61/244; B01D15/00; B01D15/08; B01D15/26; B01D15/32; B01D15/325; B01D2311/00; B01D2311/06; B01D2311/26; B01D2311/2649; B01D2311/2697
Claims Analysis
Total Claims: 49
Claim 1
Independent Claim
135 words
A process of purifying modulator material from an extract of human leukocytes, and of separating said material from substantially all fluorescamine-reactive material and from L-suppressor, comprising the steps of: (1) dialyzing said extract through a dialysis membrane having a nominal molecular weight cutoff of about 3500, thereby producing a dialysate; (2) fractionating said dialysate by gel filtration, thereby producing a plurality of dialysate-fractions; (3) modulator-assaying said dialysate-fractions, thereby ascertaining modulator activity of said dialysate-fractions; (4) selecting modulator-assayed dialysate-fractions having substantial modulator activity and pooling them; (5) applying said selected and pooled modulator-assayed dialysate-fractions to a reverse-phase high pressure liquid chromatography column, packed with octadecyl silane, (6) eluting said column with an ethanol-in water gradient, thereby producing a plurality of effluent-fractions; and (7) selecting predetermined effluent-fractions on the basis of ethanol concentration therein, and collecting them.
Claim 2
Dependent Claim
135 words
A process of purifying modulator material from an extract of human leukocytes, and of separating said material from substantially all fluorescamine-reactive material and from L-suppressor, comprising the steps of: (1) dialyzing said extract through a dialysis membrane having a nominal molecular weight cutoff of about 3500, thereby producing a dialysate; (2) fractionating said dialysate by gel filtration, thereby producing a plurality of dialysate-fractions; (3) modulator-assaying said dialysate-fractions, thereby ascertaining modulator activity of said dialysate-fractions; (4) selecting modulator-assayed dialysate-fractions having substantial modulator activity and pooling them; (5) applying said selected and pooled modulator-assayed dialysate-fractions to a reverse-phase high pressure liquid chromatography column, packed with octadecyl silane, (6) eluting said column with an ethanol-in water gradient, thereby producing a plurality of effluent-fractions; and (7) selecting predetermined effluent-fractions on the basis of ethanol concentration therein, and collecting them.
Claim 3
Dependent Claim
53 words
The process of claim 1 wherein: (a) said gradient includes ethanol in water from a concentration of approximately 0% to at least approximately 12%, and (b) said selected and collected effluent-fractions are those having a refractive index between approximately 1.332 and approximately 1.336, whereby a first amplifier is extracted in substantially purified form.
Claim 4
Dependent Claim
51 words
The process of claim 1 wherein: (a) said gradient includes ethanol in water from below approximately 28% to at least approximately 50%, and (b) said selected and collected effluent-fractions are those having a refractive index between approximately 1.340 and approximately 1.347, whereby a second amplifier is extracted in substantially purified form.
Claim 5
Dependent Claim
51 words
The process of claim 1 wherein: (a) said gradient includes ethanol in water from below approximately 65% to at least approximately 80%, and (b) said selected and collected effluent-fractions are those having a refractive index between approximately 1.353 and approximately 1.355, whereby a third amplifier is extracted in substantially purified form.
Claim 6
Dependent Claim
51 words
The process of claim 1 wherein: (a) said gradient includes ethanol in water from below approximately 81% to at least approximately 91%, and (b) said selected and collected effluent-fractions are those having a refractive index between approximately 1.356 and approximately 1.358, whereby a fourth amplifier is extracted in substantially purified form.
Claim 7
Dependent Claim
51 words
The process of claim 1 wherein: (a) said gradient includes ethanol in water from below approximately 92% to at least approximately 97%, and (b) said selected and collected effluent-fractions are those having a refractive index between approximately 1.3590 and approximately 1.3598, whereby a fifth amplifier is extracted in substantially purified form.
Claim 8
Dependent Claim
51 words
The process of claim 1 wherein: (a) said gradient includes ethanol in water from below approximately 97% to at least approximately 99%, and (b) said selected and collected effluent-fractions are those having a refractive index between approximately 1.3600 and approximately 1.3608, whereby a first suppressor is extracted in substantially purified form.
Claim 9
Dependent Claim
48 words
The process of claim 1 wherein: (a) said gradient includes ethanol in water from below approximately 98% to and including pure ethanol, and (b) said selected and collected effluent-fractions are those having a refractive index over approximately 1.3608, whereby a sixth amplifier is extracted in substantially purified form.
Claim 10
Dependent Claim
127 words
A process of purifying L-suppressor material from an extract of human leukocytes, and of separating said L-suppressor material from amplifiers 1-6, from S-suppressor, and from substantially all fluorescamine-reactive material, comprising the steps of: (1) dialyzing said extract through a first dialysis membrane having a nominal molecular weight cutoff of approximately 12,000, thereby producing a first dialysate, (2) dialyzing said first dialysate through a second dialysis membrane having a nominal molecular weight cutoff of about 3500, thereby producing a second dialysate and a retentate of the second dialysis, (3) applying said retentate of the second dialysis to a hydroxylapatite chromatography column, (4) eluting said column with ammonium bicarbonate aqueous solution gradient from approximately 0.08M to approximately 0.2M, and (5) collecting the material eluting between approximately 0.1M and 0.15M.
Claim 11
Dependent Claim
127 words
A process of purifying L-suppressor material from an extract of human leukocytes, and of separating said L-suppressor material from amplifiers 1-6, from S-suppressor, and from substantially all fluorescamine-reactive material, comprising the steps of: (1) dialyzing said extract through a first dialysis membrane having a nominal molecular weight cutoff of approximately 12,000, thereby producing a first dialysate, (2) dialyzing said first dialysate through a second dialysis membrane having a nominal molecular weight cutoff of about 3500, thereby producing a second dialysate and a retentate of the second dialysis, (3) applying said retentate of the second dialysis to a hydroxylapatite chromatography column, (4) eluting said column with ammonium bicarbonate aqueous solution gradient from approximately 0.08M to approximately 0.2M, and (5) collecting the material eluting between approximately 0.1M and 0.15M.
Claim 12
Dependent Claim
8 words
The product of the process of claim 1.
Claim 13
Dependent Claim
8 words
The product of the process of claim 2.
Claim 14
Dependent Claim
8 words
The product of the process of claim 3.
Claim 15
Dependent Claim
8 words
The product of the process of claim 4.
Claim 16
Dependent Claim
8 words
The product of the process of claim 5.
Claim 17
Dependent Claim
8 words
The product of the process of claim 6.
Claim 18
Dependent Claim
8 words
The product of the process of claim 7.
Claim 19
Dependent Claim
8 words
The product of the process of claim 8.
Claim 20
Dependent Claim
8 words
The product of the process of claim 9.
Claim 21
Dependent Claim
137 words
A composition of matter consisting of material which: (a) is derived from leukocytes; (b) is in purified form, and is purified substantially free of fluorescamine-reactive substances; (c) affects a response of the immunity system of an animal or human body, portion of said body, or matter taken therefrom, where said response: (1) is to the reintroduction to said body of at least one antigen to which said body has been previously exposed, (2) is specifically attributable to a function of the immunity system of said animal or human, and (3) occurs following said reintroduction of said antigen; and (d) is substantially free of other, different material that has an effect on said response opposite to the effect to the effect of said composition on said response-- said composition of matter being hereinafter referred to as "modulator material."
Claim 22
Dependent Claim
137 words
A composition of matter consisting of material which: (a) is derived from leukocytes; (b) is in purified form, and is purified substantially free of fluorescamine-reactive substances; (c) affects a response of the immunity system of an animal or human body, portion of said body, or matter taken therefrom, where said response: (1) is to the reintroduction to said body of at least one antigen to which said body has been previously exposed, (2) is specifically attributable to a function of the immunity system of said animal or human, and (3) occurs following said reintroduction of said antigen; and (d) is substantially free of other, different material that has an effect on said response opposite to the effect to the effect of said composition on said response-- said composition of matter being hereinafter referred to as "modulator material."
Claim 23
Dependent Claim
42 words
Modulator material of claim 19 with the further limitation that: (a) said luekocytes from which said modulator is derived are human leukocytes; and (b) said body is a human body-- said further-limited modulator material being hereinafter referred to as "human modulator material."
Claim 24
Dependent Claim
36 words
Human modulator material of claim 20 with the further limitation that said modulator affects said response by increasing the speed or magnitude of said response--said further-limited human modulator material being hereinafter referred to as "amplifier material."
Claim 25
Dependent Claim
37 words
Amplifier material of claim 21 with the further limitation that said amplifier material is: (a) dialyzable through a dialysis membrane having a nominal molecular weight cutoff of about 3500; and (b) O.S. elutable with an ethanol-in-water gradient.
Claim 26
Dependent Claim
112 words
Amplifier material of claim 22, further limited by being material substantially: (a) entirely O.S. elutable in the portions of said gradient where the effluent has a refractive index between approximately 1.332 and approximately 1.336; (b) not O.S. elutable in the portions of said gradient where the effluent has a refractive index under approximately 1.332; (c) free from material O.S. elutable in the portions of said gradient where the effluent has a refractive index under approximately 1.332; and (d) free from material O.S. elutable only in the portions of said gradient where the effluent has a refractive index over approximately 1.336-- said further limited amplifier material being hereinafter referred to as "amplifier 1."
Claim 27
Dependent Claim
54 words
Amplifier material of claim 22, further limited by being material substantially: (a) entirely O.S. elutable in the portions of said gradient where the effluent has a refractive index between approximately 1.340 and approximately 1.347; (b) not O.S. elutable in the portions of said gradient where the effluent has a refractive index under approximately 1.340;
Claim 28
Dependent Claim
58 words
(c) free from material O.S. elutable in the portions of said gradient where the effluent has a refractive index under approximately 1.340; and (d) free from material O.S. elutable only in the portions of said gradient where the effluent has a refractive index over approximately 1.347-- said further limited amplifier material being hereinafter referred to as "amplifier 2."
Claim 29
Dependent Claim
58 words
(c) free from material O.S. elutable in the portions of said gradient where the effluent has a refractive index under approximately 1.340; and (d) free from material O.S. elutable only in the portions of said gradient where the effluent has a refractive index over approximately 1.347-- said further limited amplifier material being hereinafter referred to as "amplifier 2."
Claim 30
Dependent Claim
111 words
Amplifier material of claim 22, further limited by being substantially: (a) entirely O.S. elutable in the portions of said gradient where the effluent has a refractive index between approximately 1.353 and approximately 1.355; (b) not O.S. elutable in the portions of said gradient where the effluent has a refractive index under approximately 1.353; (c) free from material O.S. elutable in the portions of said gradient where the effluent has a refractive index under approximately 1.353; and (d) free from material O.S. elutable only in the portions of said gradient where the effluent has a refractive index over approximately 1.355-- said further limited amplifier material being hereinafter referred to as "amplifier 3."
Claim 31
Dependent Claim
112 words
Amplifier material of claim 22, further limited by being substantially: (a) entirely O.S. elutable in the portions of said gradient where the effluent has a refractive index between approximately 1.356 and approximately 1.358; (b) not O.S. elutable in the portions of said gradient where the effluent has a refractive index under approximately 1.356; and (c) free from material O.S. elutable in the portions of said gradient where the effluent has a refractive index under approximately 1.356; and (d) free from material O.S. elutable only in the portions of said gradient where the effluent has a refractive index over approximately 1.358-- said further limited amplifier material being hereinafter referred to as "amplifier 4."
Claim 32
Dependent Claim
111 words
Amplifier material of claim 22, further limited by being substantially: (a) entirely O.S. elutable in the portions of said gradient where the effluent has a refractive index between approximately 1.359 and approximately 1.360; (b) not O.S. elutable in the portions of said gradient where the effluent has a refractive index under approximately 1.359; (c) free from material O.S. elutable in the portions of said gradient where the effluent has a refractive index under approximately 1.359; and (d) free from material O.S. elutable only in the portions of said gradient where the effluent has a refractive index over approximately 1.360-- said further limited amplifier material being hereinafter referred to as "amplifier 5."
Claim 33
Dependent Claim
85 words
Amplifier material of claim 22, further limited by being substantially: (a) entirely O.S. elutable in the portions of said gradient where the effluent has a refractive index over approximately 1.361; (b) not O.S. elutable in the portions of said gradient where the effluent has a refractive index under approximately 1.361; and (c) free from material O.S. elutable in the portions of said gradient where the effluent has a refractive index under approximately 1.361-- said further limited amplifier material being hereinafter referred to as "amplifier 6."
Claim 34
Dependent Claim
37 words
Human modulator material of claim 20 with the further limitation that said modulator material affects said response by decreasing the speed or magnitude of said response--said further-limited human modulator material being hereinafter referred to as "suppressor material."
Claim 35
Dependent Claim
126 words
Suppressor material of claim 29, further limited by being material: (a) dialyzable through a dialysis membrane having a nominal molecular weight cutoff of approximately 3500; (b) substantially entirely O.S. elutable in the portions of said gradient where the effluent has a refractive index between approximately 1.3600 and approximately 1.3608; (c) substantially not O.S. elutable in the portions of said gradient where the effluent has a refractive index under approximately 1.3600; (d) substantially free of material O.S. elutable in the portions of said gradient where the effluent has a refractive index under approximately 1.3600; and (e) substantially free of material O.S. elutable only in the portions of said gradient where the refractive index is over approximately 1.3608-- said further-limited suppressor material being hereinafter refrerred to as "S-suppressor."
Claim 36
Dependent Claim
106 words
Suppressor material of claim 29, further limited by being material: (a) dialyzable through a dialysis membrane having a nominal molecular weight cutoff of approximately 12,000; (b) not dialyzable through a dialysis membrane having a nominal molecular weight cutoff of approximately 3500; (c) substantially entirely chromatographically elutable from hydroxylapatite by ammonium bicarbonate aqueous solution gradient, in the portions of said gradient between approximately 0.1M and approximately 0.15M; (d) substantially not so elutable below approximately 0.1M; (e) substantially free of material so elutable below approximately 0.1M; and (f) substantially free of material so elutable only above approximately 0.15M-- said further-limited suppressor material being hereinafter referred to as "L-suppressor."
Claim 37
Dependent Claim
46 words
Human modulator material of claim 20, in an effective dosage amount, and in a pharmaceutically acceptable vehicle, thereby comprising a pharmaceutical composition for modulating the response of the immune system of a human body to reintroduction of antigens to which said body has previously been exposed.
Claim 38
Dependent Claim
25 words
The composition of claim 32 wherein said human modulator material is amplifier material and said response is modulated by increasing the speed or magnitude thereof.
Claim 39
Dependent Claim
23 words
The composition of claim 33 wherein said amplifier material includes at least one member of the group consisting of amplifiers 1 to 6.
Claim 40
Dependent Claim
25 words
The composition of claim 32 wherein said human modulator material is suppressor material and said response is modulated by decreasing the speed or magnitude thereof.
Claim 41
Dependent Claim
11 words
The composition of claim 35 wherein said suppressor material includes S-suppressor.
Claim 42
Dependent Claim
11 words
The composition of claim 35 wherein said suppressor material includes L-suppressor.
Claim 43
Independent Claim
44 words
A method of modulating the immune response of a human body to the reintroduction of antigen to which said human body has previously been exposed, comprising administering to said body human modulator material, in an effective dosage amount, and in a pharmaceutically acceptable vehicle.
Claim 44
Independent Claim
44 words
A method of modulating the immune response of a human body to the reintroduction of antigen to which said human body has previously been exposed, comprising administering to said body human modulator material, in an effective dosage amount, and in a pharmaceutically acceptable vehicle.
Claim 45
Dependent Claim
25 words
The method of claim 38 wherein said human modulator material is amplifier material and said response is modulated by increasing the speed or magnitude thereof.
Claim 46
Dependent Claim
23 words
The method of claim 39 wherein said amplifier material includes at least one member of the group consisting of amplifiers 1 to 6.
Claim 47
Dependent Claim
25 words
The method of claim 38 wherein said human modulator material is suppressor material and said response is modulated by decreasing the speed or magnitude thereof.
Claim 48
Dependent Claim
11 words
The method of claim 41 wherein said suppressor material is S-suppressor.
Claim 49
Dependent Claim
11 words
The method of claim 41 wherein said suppressor material is L-suppressor.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1987-08-28 | 1988-02-27 | 1988-08-28 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-08-28 | 1992-02-27 | 1992-08-28 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-08-28 | 1996-02-27 | 1996-08-28 | $7700.00 | Window Closed / Lapsed |