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US4486408A
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Insoluble crosslinked cytotoxic oxidase-peroxidase system
Assignee: Individual
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Filing: 1981-04-07
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Granted: 1984-12-04
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Expires: 2001-04-07
Abstract
Abstract A therapeutic preparation, and a process of using the preparation, for the control of tumor growth and immune diseases. The preparation is an immobilized cytotoxic enzyme system which is carried by an insoluble substrate that is administered into the mammalian subject by injection or implantation. The preferred embodiment of the preparation uses a polymerized albumin carrier containing immobilized glucose oxidase and peroxidase. The immobilized enzyme system is prepared in the form of small particles that can be administered either by implantation or by injection into tumor-bearing mammals. To date, antibacterial activity has been demonstrated only in vitro. Intraperitoneal injection into rats bearing Novikoff hepatomas or into mice bearing B16-F1 melanomas or L1210 (Lymphoid) tumors has resulted in the selected killing of tumor tissues. Several daily administrations are required to achieve complete destruction of the tumor. Additionally, the preparation is also active as a stimulant of the specific immune response to neoplasia. Its effect on the immune system could be useful in the stimulation of the immune system in diseases where the action of the endogenous immune system is insufficient.
Inventors
- 👤 Johnathan L. Kiel
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 08:26
CPC Classifications
A; A61; A61K; A61K38/00; A61K38/16; A61K38/43; A61K38/44; A61K38/17; A61K38/38; A61K38/385; A61K38/443; A61K47/00; A61K47/30; A61K47/42; A61K9/00; A61K9/0012; A61K9/0019; A61K9/14; A61K9/141; A61K9/146; A61K9/19; C; C12; C12N; C12N11/00; C12N11/18; C12Y; C12Y101/00; C12Y101/03; C12Y101/03004; C12Y111/00; C12Y111/01; C12Y111/01007; A61K9/06
Claims Analysis
Total Claims: 52
Claim 1
Independent Claim
91 words
A process for preparing a composition of matter possessing cytotoxic activity toward prokaryotic and eukaryotic cells comprising the steps of: a. mixing in a buffer solution: a first enzyme which is an oxidase; a second enzyme which is a peroxidase, wherein said second enzyme is capable of chemically cross-linking with said first enzyme, and wherein both enzymes remain active; and a support; b. adding a polymerizing agent to said solution; and c. allowing said solution to polymerize; and wherein the final composition of said composition of matter forms a cross-linked composition.
Claim 2
Dependent Claim
91 words
A process for preparing a composition of matter possessing cytotoxic activity toward prokaryotic and eukaryotic cells comprising the steps of: a. mixing in a buffer solution: a first enzyme which is an oxidase; a second enzyme which is a peroxidase, wherein said second enzyme is capable of chemically cross-linking with said first enzyme, and wherein both enzymes remain active; and a support; b. adding a polymerizing agent to said solution; and c. allowing said solution to polymerize; and wherein the final composition of said composition of matter forms a cross-linked composition.
Claim 3
Dependent Claim
46 words
A process, as set forth in claim 1, wherein said process comprises the additional steps of: a. cooling said polymerized solution until a gel is formed; B. removing any soluble material in said gel, thereby resulting in an insoluble gel; and c. drying said insoluble gel.
Claim 4
Dependent Claim
16 words
A process as set forth in claim 1, wherein said first enzyme is a glucose oxidase.
Claim 5
Dependent Claim
15 words
A process, as set forth in claim 1, wherein said second enzyme is horseradish peroxidase.
Claim 6
Dependent Claim
15 words
A process, as set forth in claim 1, wherein said second enzyme is lacto peroxidase.
Claim 7
Dependent Claim
15 words
A process, as set forth in claim 1, wherein said support is bovin serum albumin.
Claim 8
Dependent Claim
15 words
A process, as set forth in claim 1, wherein said support is human serum albumin.
Claim 9
Dependent Claim
29 words
A process, as set forth in claim 2, wherein said process further includes the additional steps of: a. freezing said insoluble gel; and b. lyophilizing said frozen insoluble gel.
Claim 10
Dependent Claim
68 words
A process, as set forth in claim 2, wherein said step of removing any soluble material in said gel comprises the constituent steps of: a. soaking said gel in liquid water to remove any soluble material in said gel, thereby resulting in a mixture of an insoluble gel in a liquid medium; b. filtering said mixture from said soaking to separate said insoluble gel from said liquid medium.
Claim 11
Dependent Claim
35 words
A process, as set forth in claim 8, wherein said process further includes the additional steps of: a. grinding said lyophilized insoluble gel; and b. mixing said ground lyophilized insoluble gel in a saline solution.
Claim 12
Dependent Claim
97 words
A composition of matter possessing cytotoxic activity toward prokaryotic and eukaryotic cells comprising: a. a first enzyme which is an oxidase b. a second enzyme which is a peroxidase, wherein said second enzyme is chemically cross-linked with said first enzyme, and wherein both enzymes remain active; c. a support; wherein said first enzyme, said second enzyme, and said support are mixed in a buffer solution; and d. a polymerizing agent which is added to said solution, whereby said solution is polymerized; and wherein the final composition of said composition of matter forms a cross-linked composition of matter.
Claim 13
Dependent Claim
97 words
A composition of matter possessing cytotoxic activity toward prokaryotic and eukaryotic cells comprising: a. a first enzyme which is an oxidase b. a second enzyme which is a peroxidase, wherein said second enzyme is chemically cross-linked with said first enzyme, and wherein both enzymes remain active; c. a support; wherein said first enzyme, said second enzyme, and said support are mixed in a buffer solution; and d. a polymerizing agent which is added to said solution, whereby said solution is polymerized; and wherein the final composition of said composition of matter forms a cross-linked composition of matter.
Claim 14
Dependent Claim
33 words
A composition of matter, set forth in claim 11, wherein said polymerized solution is cooled to form a gel, and the gel is dryed, and soluble material is removed from said dried gel.
Claim 15
Dependent Claim
18 words
A composition of matter, as set forth in claim 11, wherein said first enzyme is a glucose oxidase.
Claim 16
Dependent Claim
17 words
A composition of matter, as set forth in claim 11, wherein said second enzyme is horseradish peroxidase.
Claim 17
Dependent Claim
17 words
A composition of matter, as set forth in claim 11, wherein said second enzyme is lacto peroxidase.
Claim 18
Dependent Claim
17 words
A composition of matter, as set forth in claim 11, wherein said support is bovin serum albumin.
Claim 19
Dependent Claim
17 words
A composition of matter, as set forth in claim 11, wherein said support is human serum albumin.
Claim 20
Dependent Claim
51 words
A composition of matter possessing cytotoxic activity toward prokaryotic and eukaryotic cells comprising: a. a first enzyme which is an oxidase; b. a second enzyme which is a peroxidase; and c. a support; wherein said first enzyme, said second enzyme, and said support are chemically cross-linked to form an insoluble material.
Claim 21
Dependent Claim
51 words
A composition of matter possessing cytotoxic activity toward prokaryotic and eukaryotic cells comprising: a. a first enzyme which is an oxidase; b. a second enzyme which is a peroxidase; and c. a support; wherein said first enzyme, said second enzyme, and said support are chemically cross-linked to form an insoluble material.
Claim 22
Dependent Claim
17 words
A composition of matter, as set forth in claim 18, wherein said composition comprises a solid polymer.
Claim 23
Dependent Claim
18 words
A composition of matter, as set forth in claim 18, wherein said first enzyme is a glucose oxidase.
Claim 24
Dependent Claim
17 words
A composition of matter, as set forth in claim 18, wherein said second enzyme is horseradish peroxidase.
Claim 25
Dependent Claim
17 words
A composition of matter, as set forth in claim 18, wherein said second enzyme is lacto peroxidase.
Claim 26
Dependent Claim
17 words
A composition of matter, as set forth in claim 18, wherein said support is bovine serum albumin.
Claim 27
Dependent Claim
17 words
A composition of matter, as set forth in claim 18, wherein said support is human serum albumin.
Claim 28
Dependent Claim
110 words
A process for cytolysis of prokaryotic and eukaryotic cells in living organisms comprising the steps of: a. preparing a mixture of a support and a composition of matter possessing cytolysis activity toward prokaryotic and eukeryotic cells in living organisms, wherein said composition of matter is cross-linked with said support and is formed by: mixing in a buffer solution a first enzyme which is an oxidase, a second enzyme which is a peroxidase, and said support; and polymerizing said composition of matter by adding a polymerizing agent thereto such that the final composition of said composition of matter forms a cross-linked composition; and b. inserting said mixture into a living organism.
Claim 29
Dependent Claim
110 words
A process for cytolysis of prokaryotic and eukaryotic cells in living organisms comprising the steps of: a. preparing a mixture of a support and a composition of matter possessing cytolysis activity toward prokaryotic and eukeryotic cells in living organisms, wherein said composition of matter is cross-linked with said support and is formed by: mixing in a buffer solution a first enzyme which is an oxidase, a second enzyme which is a peroxidase, and said support; and polymerizing said composition of matter by adding a polymerizing agent thereto such that the final composition of said composition of matter forms a cross-linked composition; and b. inserting said mixture into a living organism.
Claim 30
Dependent Claim
69 words
A process, as set forth in claim 25, wherein after performing the step of preparing a mixture of a support and of a composition of matter and prior to performing the step of inserting said mixture into a living organism, the following additional steps are performed: a. cooling said composition of matter to form a gel; b. drying said gel; and c. removing all soluble material from said gel.
Claim 31
Dependent Claim
22 words
A process, as set forth in claim 25, wherein said living organism is selected from the group of animals known as mammals.
Claim 32
Dependent Claim
14 words
A process, as set forth in claim 25, wherein said mammal is a human.
Claim 33
Dependent Claim
16 words
A process, as set forth in claim 25, wherein said cytolytic activity is directed against neoplasia.
Claim 34
Dependent Claim
46 words
A process, as set forth in claim 25, wherein the step of inserting said mixture into a living organism is performed by infusing said mixture into said living organism, such that said infused mixture stimulates the activity of the endogeneous immune system of said living organism.
Claim 35
Dependent Claim
73 words
A composition of matter possessing cytotoxic activity toward prokaryotic and eukaryotic cells comprising: a. a peroxidase selected from the group consisting of horseradish peroxidase, lactoperoxidase, myeloperoxidase and snake skin peroxidase; b. an enzyme that produces hydrogen peroxide or superoxide in sufficient amounts and for a sufficient period of time as to maintain the cytotoxicity of said peroxidase; and c. a support; wherein said peroxidase, said enzyme, and said support form a cross-linked composition.
Claim 36
Dependent Claim
73 words
A composition of matter possessing cytotoxic activity toward prokaryotic and eukaryotic cells comprising: a. a peroxidase selected from the group consisting of horseradish peroxidase, lactoperoxidase, myeloperoxidase and snake skin peroxidase; b. an enzyme that produces hydrogen peroxide or superoxide in sufficient amounts and for a sufficient period of time as to maintain the cytotoxicity of said peroxidase; and c. a support; wherein said peroxidase, said enzyme, and said support form a cross-linked composition.
Claim 37
Dependent Claim
16 words
A composition of matter, as set forth in claim 31, wherein said support comprises a protein.
Claim 38
Dependent Claim
15 words
A composition of matter, as set forth in claim 32, wherein said protein is albumin.
Claim 39
Dependent Claim
16 words
A composition of matter, as set forth in claim 31, wherein said enzyme is glucose oxidase.
Claim 40
Dependent Claim
16 words
A composition of matter, as set forth in claim 31, wherein said enzyme is xanthine oxidase.
Claim 41
Dependent Claim
38 words
A composition of matter, as set forth in claim 31, wherein said peroxidase, said enzyme, and said support are dissolved in a buffer solution, and a polymerizing agent is added to said solution, whereby said solution is polymerized.
Claim 42
Dependent Claim
34 words
A composition of matter, as set forth in claim 36, wherein said polymerized solution is cooled to form a gel, and the gel is dried, and soluble material is removed from said dried gel.
Claim 43
Dependent Claim
99 words
A solid composition of matter possessing cytotoxic activity against tumor cells upon administration to a living tumor-bearing subject comprising: a. a first enzyme selected from the group of enzymes which in the presence of hydrogen peroxide and a halide ion possess cytolytic activity toward prokaryotic and eukaryotic cells; b. a second enzyme selected from the group consisting of enzymes that produce hydrogen peroxide or superoxide in sufficient amounts and for a sufficient period of time as to maintain the cytotoxicity of said first enzyme; and c. a support wherein said first and second enzymes are crosslinked to said support.
Claim 44
Dependent Claim
99 words
A solid composition of matter possessing cytotoxic activity against tumor cells upon administration to a living tumor-bearing subject comprising: a. a first enzyme selected from the group of enzymes which in the presence of hydrogen peroxide and a halide ion possess cytolytic activity toward prokaryotic and eukaryotic cells; b. a second enzyme selected from the group consisting of enzymes that produce hydrogen peroxide or superoxide in sufficient amounts and for a sufficient period of time as to maintain the cytotoxicity of said first enzyme; and c. a support wherein said first and second enzymes are crosslinked to said support.
Claim 45
Dependent Claim
17 words
A composition of matter, as set forth in claim 38, wherein said first enzyme is horseradish peroxidase.
Claim 46
Dependent Claim
16 words
A composition of matter, as set forth in claim 38, wherein said first enzyme is lactoperoxidase.
Claim 47
Dependent Claim
16 words
A composition of matter, as set forth in claim 38, wherein said first enzyme is myeloperoxidase.
Claim 48
Dependent Claim
18 words
A composition of matter, as set forth in claim 38, wherein said first enzyme is snake skin peroxidase.
Claim 49
Dependent Claim
16 words
A composition of matter, as set forth in claim 38, wherein said first enzyme is peroxidase.
Claim 50
Dependent Claim
17 words
A composition of matter, as set forth in claim 38, wherein said second enzyme is glucose oxidase.
Claim 51
Dependent Claim
16 words
A composition of matter, as set forth in claim 38, wherein said support is a protein.
Claim 52
Dependent Claim
15 words
A composition of matter, as set forth in claim 45, wherein said protein is albumin.
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) | 1987-12-04 | 1988-06-04 | 1988-12-04 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-12-04 | 1992-06-04 | 1992-12-04 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-12-04 | 1996-06-04 | 1996-12-04 | $7700.00 | Window Closed / Lapsed |