Therapeutic apheresis filter system combination consisting of a separator filter, an L-tryptophan TSO bioreactor, and a fractionator.

Abstract

Filter system for filtering plasma with a separator and a fractionator and a bioreactor connected between the separator and the fractionator, which contains silica and immobilized tryptophan side chain oxidase (TSO) as the enzyme.

Description

Specification of technical area

[0001] Filter systems for therapeutic apheresis as supportive treatment in cancer therapies State of the art and deficiencies in current designs

[0002] One of the major obstacles in modern cancer therapy is the development of resistance to chemotherapeutic agents and new targeted therapies such as kinase inhibitors and monoclonal antibodies (1, 2). This resistance makes it difficult to sustainably control tumor growth. Furthermore, there is currently no single therapeutic intervention that is effective for all types of cancer. Existing options are often associated with severe side effects, which severely impair patients' quality of life (2). Due to these limitations, there is an urgent need for new therapeutic concepts that are both more effective and better tolerated. In particular, the following sources (1-15) are referred to: 1. Reichle A, Diddens H, Rastetter J, Berdel WE (1991). Resistance mechanisms of malignant cells to cytostatic drugs. Dtsch Med Wschr 116:186–191. 2. Volm M, Mattern J, Samsel B (1991). Increased incidence of cytostatic-resistant lung tumors in smokers. Dtsch Med Wschr 116:1303-1306. 3. Rosenfeld HJ, Watanabe AK, Roberts J(1977). Mechanism of action of indolyl-3-alkane-α-hydroxylase. Biol Chem 252:6970–6973. 4. Roberts J Schmid FA, Rosenfeld HJ (1979). Biologic and antineoplastic effects of enzyme-mediated in vivo depletion of L-glutamine, L-tryptophan and L.-histidine. Cancer Treat Rep 63:1045–1054. 5. Cook SJ, Pogson CI, Smith SA (1980). Indoleamine 2,3dioxygenase. Biochem J 189:461–466, 6. Kidd JG (1953). Regression of transplanted lymphoma induced in vive by means of normal Guinea pigserum: II. Studies on the nature of the active serum constituent: histological mechanism of regression: tests for effects of guinea pig serum on lymphoma cells in vitro: Discussion. J Exp Med 98:565. 7. Kidd JG (1953). Regression of transplanted lymphoma induced in vivo by means of normal Guinea pigserum: II. Studies on the nature of the active serum constituent histological mechanism of regression: tests for effects of guinea pig serum on lymphoma cell in vitro: Discussion. J Exp Med 98:583. 8. Broome JD (1961). Evidence that the L-asparaginase activity in guinea pig serum is responsible for its antilymphoma effects. Nature 191:114. 9. Schmer G, Roberts J (1979). Molecular engineering of the L-tryptophandepleting enzymeindolyl-3-alkane-αhydroxylase. Canc Treat Rep 63:1123-1126. 10. Schmer G, Roberts J (1979). Induction of hypothermia in mice by semiartificial cell containing indolyl-3-alkane-αhydroxylase. Trans Am Soc Artif Int Org 25:39-43. 11. Dennis MB, Jensen WH, Bauermeister U Vienken J, Schmer G (1988). Successful use of a miniturized plasmapheresis circuit for long-term use in rabbits. Trans Am Soc Artif Int Org. 34:651-654. 12. Bambauer R, Yefu W. (2023). L-Tryptophan Depletion Using a New Bioreactor: A Possible New Cancer Therapy. Br J Healthc Med Res. 10(1):120-130. Doi:10.14738 / jbe mi.101.13737. 13. Schmer G, Kruegger M, Cole JJ (1977). Gel-bound resealed red cell membranes: A new type of semiartificial organs. J Biol Chem 252:2640-2647. 14. Straube R, Müller G, Voit-Bak K, et al. (2019). Metabolic and nonmetabolic peripheral neuropathy: Is there a place for therapeutic apheresis? Hormone Metab Res. 51(12):779- 784. 15. Bornstein SR, Voit-Bak K, Rosenthal P, et al. (2020). Extracorporeal apheresis therapy for Alzheimer disease-targeting lipids, stress, and inflammation. Molec Psych. 25:275-282. Technical problem and its solution

[0003] Recent studies have shown that extracorporeal systems with antitumor enzymes, so-called bioreactors, offer a promising option for tumor treatment (3-5). One such approach focuses on the depletion of essential amino acids, particularly L-tryptophan, which is used by tumor cells for protein synthesis and survival (6-8).

[0004] By removing L-tryptophan, tumor growth can be significantly reduced and even stopped (9). A key advantage of this strategy is that L-tryptophan is an essential amino acid that cannot be synthesized by the tumor itself, unlike other amino acids such as L-asparagine, which the tumor can replace through de novo synthesis (9). This prevents the development of resistance to treatment and makes L-tryptophan an ideal target for extracorporeal depletion.

[0005] For this purpose, the enzyme tryptophan side-chain oxidase (TSO) is used, which can degrade L-tryptophan in blood plasma. There are three isoenzymes: TSO I, II, and III, with TSO III being the most efficient in degrading L-tryptophan (10). A key problem, however, is that these enzymes, when derived from bacterial sources, can be contaminated with high levels of endotoxins that have toxic effects on the human body (11). To circumvent this, this invention relies on endotoxin-free TSO enzymes from fungal sources (11, 12).

[0006] To solve these problems, the features of the independent claims are proposed. Advantageous further developments are set forth in the dependent claims.

[0007] A filter system for filtering plasma may comprise a separator (primary filter) and a fractionator (secondary filter), and at least one bioreactor connected between the separator and the fractionator. The bioreactor may contain silica and immobilized tryptophan side-chain oxidase (TSO) as the enzyme.

[0008] The TSO can advantageously be a TSO I isoenzyme, preferably a TSO II isoenzyme, and particularly preferably a TSO III isoenzyme. The TSO can preferably be an endotoxin-free TSO from fungal sources.

[0009] The fractionator can be a highly selective filter system made of biocompatible hollow-fiber membranes that removes specific molecules from the plasma, especially those with a molecular size of more than 10 nm. The bioreactor can contain silica spheres coupled to the TSO enzymes. The TSO enzymes can be coupled to Zetaffinity microcolumns. The Zetaffinity microcolumns can preferably be made of polyacrylic cellulose copolymer.

[0010] A first pump can be connected upstream of the bioreactor and / or a second pump can be connected downstream of the bioreactor. A first expansion chamber can be provided upstream of the first pump and / or a second expansion chamber can be provided downstream of the second pump.

[0011] An apheresis device can have a filter system, particularly for extracorporeal plasmapheresis. The filter system can comprise at least one separator and one fractionator, wherein the fractionator can be a highly selective filter system that removes specific molecules from the plasma, particularly those with a molecular size of more than 10 nm. In addition, at least one (or more) tryptophan side chain oxidase (TSO) bioreactors can be provided, wherein the TSO bioreactor can be based on activated silica, preferably silica spheres, coupled with immobilized tryptophan side chain oxidase enzymes.

[0012] Methods for performing apheresis can be proposed, in particular extracorporeal plasmapheresis with double-membrane filtration, wherein withdrawn whole blood is passed through a filter system. The filter system can comprise a separator and a fractionator, wherein the separator separates plasma from solid blood components of the whole blood. The fractionator can be a highly selective filter system that removes specific molecules from the plasma, in particular those with a molecular size of more than 10 nm. At least one tryptophan side chain oxidase (TSO) bioreactor can be provided, and the TSO bioreactor can be based on activated silica, preferably silica spheres, coupled with immobilized tryptophan side chain oxidase enzymes. The plasma of the withdrawn whole blood can become purified plasma by passing through the filter system, which in turn can be combined to form whole blood.

[0013] Advantageous embodiments and further details of the present invention are described below using various exemplary embodiments with reference to schematic figures. The connection is explained in more detail in the schematic drawings. Explanation of the invention with application example

[0014] The invention describes an innovative therapeutic apheresis filter system combination, with a fractionator (TKM-58) filter and an L-tryptophan TSO bioreactor, as in Fig.1. The bioreactor is based on activated silica spheres coupled with TSO enzymes, which in turn are tightly coupled to Zetaffinity microcolumns made of polyacrylic cellulose copolymer. The TSO bioreactor is integrated into the plasmapheresis system, after the membrane that separates blood into blood cells and plasma. The plasma is passed through the bioreactor, where L-tryptophan is broken down into metabolites through enzymatic reactions. The filtered plasma is then reunited with the blood cells and returned to the patient, where the metabolites are excreted by the human kidney (13).

[0015] Zetaffinity microcolumns are specialized chromatographic columns used in protein analysis, biomolecule separation, and affinity chromatography. The name is derived from the terms "zeta" (often referring to charge effects) and "affinity" (specific interactions). They can be used for the separation and purification of proteins, peptides, and other biomolecules.

[0016] The treatment cycle, which consists of 15–20 sessions (5x / week for 4–5 hours) and can be repeated every 2–3 months, keeps the L-tryptophan level in the patient's blood consistently low, forcing the tumor to release its intracellular L-tryptophan (13). This effectively suppresses tumor growth without the serious side effects that might occur with longer treatments. Plasma filtration itself is highly efficient, and potential side effects, such as immunological reactions, can be controlled through regular plasmapheresis (14, 15). Furthermore, the disclosure in the "American Journal of Biomedical Science & Research" entitled "A New Bioreactor For L-Tryptophan Depletion as A New Cancer Therapy," ISSN: 2642-1747, pages 148–154 (12 / 2024), is incorporated in its entirety by reference into the present application.

[0017] Existing features, components, and specific details may be exchanged and / or combined to create further embodiments, depending on the required application. Any modifications within the scope of the knowledge of the person skilled in the art are implicitly disclosed in this description. New advantages through invention

[0018] This novel filter system combination offers several significant advantages. First, the integration of the TSO bioreactor with the fractionator (specifically, the TKM-58) filter enables simultaneous reduction of L-tryptophan and removal of cellular debris and toxins, increasing treatment efficiency and minimizing side effects. Second, the risk of toxic effects from endotoxins is avoided by using TSO enzymes from fungal sources that are endotoxin-free (8). Third, the integration of a pump upstream or downstream of the bioreactor, as well as expansion chambers upstream of the plasma pump and upstream of the confluence of the refluxing blood, minimizes the risk of air bubble formation in the tubing system, thus increasing the safety of the therapy.

[0019] Overall, this invention offers a promising new method for treating cancer based on the targeted depletion of an essential amino acid while ensuring high efficiency and safety through its combination of bioreactor and filter system. QUOTES CONTAINED IN THE DESCRIPTION

[0000] This list of documents submitted by the applicant was generated automatically and is included solely for the convenience of the reader. This list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions. Cited non-patent literature

[0000] Reichle A, Diddens H, Rastetter J, Berdel WE (1991). Resistance mechanisms of malignant cells to cytostatic drugs. Dtsch Med Wschr 116:186-191

[0002] Volm M, Mattern J, Samsel B (1991). Häufung von zytostatica-resistenten Lungentumoren bei Rauchern. Dtsch Med Wschr 116:1303 -1306

[0002] Rosenfeld HJ, Watanabe AK, Roberts J(1977). Mechanism of action of indolyl-3-alkane-α- hydroxylase. Biol Chem 252:6970-6973

[0002] Roberts J Schmid FA, Rosenfeld HJ (1979). Biologic and antineoplastic effects of enzyme-mediated in vivo depletion of L-glutamine, L-tryptophan and L.- histidine. Cancer Treat Rep 63:1045-1054

[0002] Cook SJ, Pogson CI, Smith SA (1980). Indoleamine 2,3dioxygenase. Biochem J 189:461- 466

[0002] Kidd JG (1953). Regression of transplanted lymphoma induced in vive by means of normal Guinea pigserum: II. Studies on the nature of the active serum constituent: histological mechanism of regression: tests for effects of guinea pig serum on lymphoma cells in vitro: Discussion. J Exp Med 98:565

[0002] Kidd JG (1953). Regression of transplanted lymphoma induced in vivo by means of normal Guinea pigserum: II. Studies on the nature of the active serum constituent histological mechanism of regression: tests for effects of guinea pig serum on lymphoma cell in vitro: Discussion. J Exp Med 98:583

[0002] Broome JD (1961). Evidence that the L-asparaginase activity in guinea pig serum is responsible for its antilymphoma effects. Nature 191:114

[0002] Schmer G, Roberts J (1979). Molecular engineering of the L-tryptophandepleting enzymeindolyl-3-alkane-αhydroxylase. Canc Treat Rep 63:1123-1126

[0002] Schmer G, Roberts J (1979). Induction of hypothermia in mice by semiartificial cell containing indolyl-3-alkane-αhydroxylase. Trans Am Soc Artif Int Org 25:39-43

[0002] Dennis MB, Jensen WH, Bauermeister U Vienken J, Schmer G (1988). Successful use of a miniturized plasmapheresis circuit for long-term use in rabbits. Trans Am Soc Artif Int Org. 34:651-654

[0002] Bambauer R, Yefu W. (2023). L-Tryptophan Depletion Using a New Bioreactor: A Possible New Cancer Therapy. Br J Healthc Med Res. 10(1):120-130. Doi:10.14738 / jbe mi.101.13737

[0002] Schmer G, Kruegger M, Cole JJ (1977). Gel-bound resealed red cell membranes: A new type of semiartificial organs. J Biol Chem 252:2640-2647

[0002] Straube R, Müller G, Voit-Bak K, et al. (2019). Metabolic and nonmetabolic peripheral neuropathy: Is there a place for therapeutic apheresis? Hormone Metab Res. 51(12):779- 784

[0002] Bornstein SR, Voit-Bak K, Rosenthal P, et al. (2020). Extracorporeal apheresis therapy for Alzheimer disease-targeting lipids, stress, and inflammation. Molec Psysch. 25:275-282

[0002] American Journal of Biomedical Science & Research“ mit dem Titel „A New Bioreactor For L-Tryptophan Depletion as A New Cancer Therapy“, ISSN: 2642-1747 auf Seite 148-154 (12 / 2024

[0016]

Claims

[1] Filter system for the filtration of plasma with a separator and a fractionator and a bioreactor connected between the separator and the fractionator, which has silica and immobilized tryptophan side chain oxidase (TSO) as enzyme. [2] Filter system according to claim 1, wherein the TSO is an isoenzyme TSO I, preferably an isoenzyme TSO II and particularly preferably an isoenzyme TSO III. [3] A filter system according to claim 1 or 2, wherein the TSO is endotoxin-free TSO from fungal sources. [4] Filter system according to one of the preceding claims, wherein the fractionator is a highly selective filter system that removes specific molecules from the plasma, in particular those with a molecular size of more than 10 nm. [5] Filter system according to one of the preceding claims, wherein the bioreactor comprises silica spheres coupled with the TSO enzymes. [6] The filter system of claim 5, wherein the TSO enzymes are coupled to Zetaffinity microcolumns. [7] The filter system of claim 6, wherein the Zetaffinity microcolumns are made of polyacrylic cellulose copolymer. [8] Filter system according to one of the preceding claims, wherein a first pump is provided upstream of the bioreactor and / or a second pump is provided downstream of the bioreactor. [9] Filter system according to one of the preceding claims, wherein a first expansion chamber is provided upstream of the first pump and / or a second expansion chamber is provided downstream of the second pump. [10] Device for cancer therapy with a plasmapheresis system for separating blood into blood cells and plasma and introducing them into separate channels and a filter system according to one of the preceding claims, and with a supply line for blood from a collection point on the patient to the plasmapheresis system, wherein the filter system is arranged downstream of the plasmapheresis system and is connected to the channel with the plasma, and with a merging of the plasma from the filter system and the separated blood cells, which flows into a channel that carries blood for introduction into the patient's blood system. [11] Apheresis device with a filter system, in particular for extracorporeal plasmapheresis, the filter system comprising: a separator and a fractionator, wherein the fractionator is a highly selective filter system that removes specific molecules from the plasma, in particular those with a molecular size of more than 10 nm; and at least one tryptophan side chain oxidase (TSO) bioreactor; wherein the TSO bioreactor is based on activated silica, preferably silica spheres, coupled with immobilized tryptophan side chain oxidase enzymes. [12] The device according to claim 11, wherein the bioreactor is arranged between the separator and the fractionator with a pump. [13] The device according to claim 11 or 12, wherein the TSO bioreactor is based on activated silica spheres coupled with TSO enzymes, which are tightly coupled to Zetaffinity microcolumns, and which consist at least partially, preferably completely, of polyacrylic cellulose copolymer. [14] The device according to at least one of the preceding claims, wherein at least one pump is arranged before or after the bioreactor and wherein the pump is in particular a plasma pump. [15] The device according to at least one of the preceding claims, wherein a first expansion chamber is provided, which is preferably arranged upstream of the pump in order to avoid air in the tube system of the apheresis device and / or wherein a second expansion chamber is provided, which is arranged upstream of a confluence of the returning blood and the filtered plasma. [16] The device according to at least one of the preceding claims, wherein the separator is a plasma filter and the fractionator is a differential filter for filtering out eluate from the plasma.

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