WO2023210691A1 - Dispositif de traitement à l'aide une solution contenant des principes actifs et système de circulation extracorporelle d'une solution contenant des principes actifs - Google Patents

Dispositif de traitement à l'aide une solution contenant des principes actifs et système de circulation extracorporelle d'une solution contenant des principes actifs Download PDF

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Publication number
WO2023210691A1
WO2023210691A1 PCT/JP2023/016454 JP2023016454W WO2023210691A1 WO 2023210691 A1 WO2023210691 A1 WO 2023210691A1 JP 2023016454 W JP2023016454 W JP 2023016454W WO 2023210691 A1 WO2023210691 A1 WO 2023210691A1
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WO
WIPO (PCT)
Prior art keywords
active ingredient
containing solution
cells
blood
processing device
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PCT/JP2023/016454
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English (en)
Japanese (ja)
Inventor
昌治 竹内
淳 澤山
文智 小沢
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国立大学法人東京大学
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Publication of WO2023210691A1 publication Critical patent/WO2023210691A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M3/00—Tissue, human, animal or plant cell, or virus culture apparatus
    • C12M3/06—Tissue, human, animal or plant cell, or virus culture apparatus with filtration, ultrafiltration, inverse osmosis or dialysis means

Definitions

  • Patent Document 2 discloses a configuration that performs a process of imparting nutritional substances to blood via hollow fibers as an extracorporeal active ingredient-containing solution circulation system.
  • the active ingredient-containing solution processing device and the extracorporeal active ingredient-containing solution circulation system of Patent Document 1 use a blood component regulator in addition to a blood purification device, a problem arises in that the device becomes larger.
  • the active ingredient-containing solution processing device and the extracorporeal active ingredient-containing solution circulation system of Patent Document 2 are provided with a nutritional substance supply channel and a body fluid supply channel in addition to the blood circulation channel, so they are different from Patent Document 1. Similarly, a problem arises in that the device becomes larger. Furthermore, as the device becomes larger, there is also the problem that the amount of blood to be circulated outside the body increases.
  • the present invention has been made in consideration of the above points, and provides an active ingredient-containing solution processing device and an extracorporeal active ingredient-containing solution circulation system that can reduce the size of the device and suppress the amount of active ingredient-containing solution to be circulated outside the body.
  • the purpose is to provide
  • the active ingredient-containing solution processing device of the first aspect and a pump that circulates at least the component in the active ingredient-containing solution from the introduction part to the discharge part of the chamber are provided.
  • An extracorporeal active ingredient-containing solution circulation system is provided.
  • FIG. 1 is a diagram showing a schematic configuration of an extracorporeal blood circulation system 100.
  • the extracorporeal blood circulation system 100 circulates at least some components of the blood outside the body of the living body LB, and performs predetermined processing on some of the blood components in the blood processing device 1.
  • the extracorporeal blood circulation system 100 includes a blood processing device 1, an infusion reservoir 10, a first introduction piping system 11, a first discharge piping system 12, a second introduction piping system 13, and a second discharge piping system 14. , a first pump 21, a second pump 22, a third pump 23, and a fourth pump 24.
  • the infusion fluid reservoir 10 stores infusion fluid.
  • the second introduction piping system 13 includes piping that introduces the infusion discharged from the infusion reservoir 10 into the blood processing apparatus 1 .
  • the second discharge piping system 14 includes piping that discharges the infusion fluid discharged from the blood processing device 1 toward the infusion fluid reservoir 10 .
  • the third pump 23 is provided in the second introduction piping system 13 and sends the transfusion toward the blood processing apparatus 1 .
  • the fourth pump 24 is provided in the second discharge piping system 14 and sends the infusion from the blood processing device 1 toward the infusion reservoir 10 .
  • the blood processing device 1 includes a chamber 30 and a linear member F.
  • the chamber 30 includes a chamber body 31, a lid 40, a fiber support (support) 50, and filters 61 and 62.
  • the chamber body 31 has a cylindrical shape extending in the vertical direction.
  • the chamber body 31 is open at the top.
  • the chamber body 31 includes a bottom wall 32 , a peripheral wall 33 , a top wall 34 , and a processing space 35 .
  • the pedestal portion 51 is located at the lower end of the fiber support 50.
  • the pedestal portion 51 has a flange shape extending in the radial direction.
  • the pedestal portion 51 has an annular magnet 51A facing downward.
  • the support shaft 52 has a cylindrical shape extending upward from the pedestal portion 51.
  • the diameter of the support shaft 52 is smaller than the diameter of the pedestal portion 51.
  • a linear member F (details will be described later) is wound around the outer periphery of the support shaft 52. As shown in FIG.
  • the fitting convex portion 53 has a cylindrical shape extending upward from the upper end of the support shaft 52.
  • the diameter of the fitting convex portion 53 is smaller than the diameter of the support shaft 52.
  • the fitting protrusion 53 is inserted into and fitted into the fitting recess 40a of the lid 40 from below.
  • the fitting protrusion 53 is removable from the fitting recess 40a.
  • the fitting recess 54 is recessed upward from the lower surface of the pedestal 51 .
  • the fitting protrusion 32B of the bottom wall portion 32 is inserted into the fitting recess 54 from below and fitted therein.
  • the through channel 55 passes through the fiber support 50 in the vertical direction.
  • the lower end of the through passage 55 opens into the fitting recess 54 .
  • the fitting recess 54 is removable from the fitting protrusion 32B.
  • the fiber support 50 When the fitting protrusion 53 fits into the fitting recess 40a of the lid 40, the fiber support 50 is positioned on the lid 40, and the through passage 55 communicates with the through passage 40b.
  • the fitting protrusion 32B fits into the fitting recess 54 the fiber support 50 is positioned in the chamber body 31, and the through passage 55 communicates with the introduction port 32D. That is, the fiber support 50 is detachably attached to the chamber 30.
  • the introduction port 32D, the through channel 55, and the through channel 40b communicate with each other.
  • the blood processing apparatus 1 in which the fiber support 50 is arranged in the processing space 35 of the chamber body 31 has a maximum diameter of about 2 cm and a maximum length of about 10 cm.
  • the filter 61 is arranged at the boundary 60A between the through-flow path 55 and the through-flow path 40b.
  • Boundary 60A is a discharge part from which blood is discharged in chamber 30.
  • the filter 62 is arranged at a boundary 60B between the introduction port 32D and the through flow path 55.
  • Boundary 60B is an introduction part into which blood is introduced into chamber 30.
  • the filters 61 and 62 stop the linear member F inside the chamber 30.
  • the mesh size of the filters 61 and 62 is smaller than the size of the linear member F.
  • the filter 62 also at the boundary 60B, which is the introduction part, in consideration of causing the liquid to flow back when performing air removal or the like.
  • the core part 210 has a hydrogel filled with a plurality of cells 115.
  • Shell portion 220 covers the outer periphery of core portion 210 .
  • the shell portion 220 is a hydrogel that is dissociable from the hydrogel forming the core portion 210 .
  • FIG. 4 is a schematic diagram illustrating the manufacturing process of cell fiber F.
  • the microfluidic device 140 includes an introduction port 110 for introducing a first liquid material to form a core section 210, an introduction port 120 for introducing a second liquid material for forming a shell section 220, and a shell section 220. It has an introduction port 130 for introducing the third liquid material to be gelled.
  • extracellular matrix components include, but are not limited to, collagen (type I, type II, type III, type V, type XI, etc.), mouse EHS tumor extract (type IV collagen, laminin, heparan sulfate proteoglycan, etc.). (including) reconstituted basement membrane components, gelatin, agar, agarose, fibrin, glycosaminoglycans, hyaluronic acid, proteoglycans, thrombin, aprotinin, alginic acid, and the like.
  • pH-responsive gelling solution examples include an alginate solution, a chitosan solution, a carboxymethyl cellulose solution, and an acrylic acid-based synthetic solution.
  • the injection speed of the solution at the introduction ports 110 and 120 is not particularly limited, but if the diameter of the microfluidic device 140 is about 50 ⁇ m to 2 mm, it may be about 10 to 500 ⁇ L/min. By adjusting the injection speed of the solution at the inlets 110 and 120, the diameter of the core portion and the coating thickness of the shell portion can be adjusted as appropriate.
  • the injection speed of the solution at the introduction port 130 is not particularly limited, but may be, for example, about 1 to 10 mL/min.
  • the core portion 210 contains various growth factors suitable for maintaining, proliferating, or expressing functions of the cells 115, such as epidermal growth factor (EGF), platelet-derived growth factor (PDGF), transforming growth factor (TGF), insulin-like Growth factors (IGF), fibroblast growth factors (FGF), nerve growth factors (NGF), etc. may be included.
  • EGF epidermal growth factor
  • PDGF platelet-derived growth factor
  • TGF transforming growth factor
  • IGF insulin-like Growth factors
  • FGF fibroblast growth factors
  • NGF nerve growth factors
  • an appropriate concentration can be selected depending on the type of growth factor.
  • the amount of blood to be used can be adjusted using a small device. becomes possible to suppress.
  • the polysaccharide is at least one selected from the group consisting of alginic acid, starch, glycogen, cellulose, xanthan gum, hyaluronic acid, carrageenan, pectin, pullulan, and salts thereof.
  • the hydrophilic polymers include polyethylene glycol, polyethyleneimine, polyvinyl alcohol, polyacrylic acid, polyacrylamide, polyvinyl alcohol, polyvinylpyrrolidone, polyvinylacetamide, polyamine, poly(4-styrene sulfonic acid), poly(allylamine hydrochloride), At least one selected from the group consisting of poly(vinylsulfonic acid, sodium salt), poly(diallyldimethylammonium chloride), and poly(2-methacryloyloxyethylphosphorylcholine).
  • a cell fiber F is obtained in which the core portion 210 is occupied by cells 115 and the shell portion 220 is gelled.
  • the shell part 220 contains the block copolymer, so by adjusting the temperature to a temperature below LCST, the block copolymer becomes water-soluble, and by stirring the cell fiber F in water, the block copolymer becomes water-soluble. , block copolymers can be removed.
  • a cellular fiber F in which the core portion 210 is filled with cells and the shell portion 220 is macroporous (porous) is obtained.
  • FIG. 5 is a fluorescence image at 37° C. of a cell fiber F having a shell portion 220 with a macroporous structure in which the core portion is filled with fluorescent nanoparticles with a diameter of 100 nm.
  • FIG. 6 is a fluorescence image of the cell fiber F in FIG. 5 when the temperature reached 4° C. after 1 hour. As shown in FIG. 5, it was confirmed that when the shell portion was at 37° C., the fluorescent nanoparticles with a diameter of 100 nm could not pass through the shell portion and remained in the core portion. As shown in FIG. 6, when the shell portion was at 4° C., no fluorescent nanoparticles with a diameter of 100 nm were observed in the core portion, and it was confirmed that the fluorescent nanoparticles passed through the shell portion.
  • S protein spike protein
  • S protein beads that can mimic the cell infection mechanism of SARS-CoV-2 will be produced.
  • S protein beads bound by the affinity of Ni-NTA and His tag were prepared.
  • the binding between the red fluorescent beads and the S protein was confirmed by measuring the fluorescence resonance energy transfer from the S protein to the red fluorescent beads.
  • a rat was used as a living LB in the extracorporeal blood circulation system 100, the first introduction piping system 11 and the first discharge piping system 12 were connected to the rat's vein, and a solution containing 4 ⁇ g/mL of S protein beads was injected into the vein. .
  • the amount of S protein beads was measured in the first introduction piping system 11 for 45 minutes every 5 minutes after the solution containing S protein beads was injected. Injection of a solution containing S protein beads and measurement of S protein beads every 5 minutes were performed for a sample in which cell fiber F was filled with VeroE6 cells and a sample in which cell fiber F was not filled with cells. .
  • Figure 8 shows the relationship between the time after injection of a solution containing S protein beads and the measured amount of S protein beads for a sample filled with VeroE6 cells and a sample in which cell fiber F is not filled with cells.
  • FIG. 8 it was confirmed that in the sample using VeroE6 cells, S protein beads decreased more over time than in the sample not using cells. This is because membrane proteins in VeroE6 cells come into contact with S protein beads and are instantly bound (infected) and captured, reducing the amount of S protein beads contained in the blood in the biological LB and purifying the blood. This is thought to be the result of
  • FIG. 9 is a fluorescence image of a sample in which cell fiber F is not filled with cells.
  • FIG. 10 is a fluorescence image of a sample in which cell fiber F was filled with VeroE6 cells. As shown in FIG. 9, in the sample in which cell fiber F was not filled with cells, capture of S protein beads was not confirmed. On the other hand, as shown in FIG. 10, in the sample in which the cell fiber F was filled with VeroE6 cells, blood purification due to the capture of S protein beads was confirmed.
  • the cell fiber F which is filled with affinity cells that have an affinity for viruses and has a length of several tens of meters, is used, so that the virus can be captured without reducing the removal ability. can do.
  • Physiologically active substances released into circulating blood include albumin, proteins such as enzymes, hormones such as insulin, exosomes, MicroRNA, sugars, lipids, and the like.
  • Exocrine cells that release albumin, a physiologically active substance, into circulating blood as an exocrine secretion include hepatocytes.
  • Exocrine cells that release enzymes, which are physiologically active substances, into circulating blood as exocrine secretions include enzyme-producing cells.
  • Exocrine cells that release hormones such as insulin, which are physiologically active substances, into circulating blood as exocrine secretions include hormone-producing cells such as ⁇ cells.
  • Exocrine cells that release exosomes, which are physiologically active substances, into the circulating blood include airway epithelial cells.
  • physiologically active substances can be replenished into the circulating blood.
  • blood exhibiting hypoalbuminemia etc. can be supplemented with albumin
  • blood exhibiting lysosomal diseases etc. can be supplemented with enzymes. That is, the extracorporeal blood circulation system 100 of this embodiment functions as a physiologically active substance. According to this embodiment, there is no need to separately provide a physiologically active substance supply channel in addition to a blood circulation channel, and it is possible to downsize the device and suppress the amount of blood to be circulated outside the body.
  • human albumin was detected in all four samples. Since human albumin is normally not detected in rats, it was confirmed that human albumin released from exocrine cells in cell fiber F was replenished into circulating blood via cell fiber F.
  • the cell fibers F can be used to connect affinity cells that express a membrane protein that has an affinity for a specific substance contained in blood, and exocrine cells that release physiologically active substances as exocrine substances into the blood.
  • the cell fiber F may include both the above-mentioned affinity cells and exocrine cells. In this case, by using the extracorporeal blood circulation system 100, both blood purification processing and physiologically active substance replenishment processing can be performed.
  • the extracorporeal blood circulation system 100 has a configuration in which the infusion circulation path is provided, but a configuration in which the infusion circulation path is not provided may also be used.
  • a configuration may be adopted in which the first discharge piping system 12 provided with the second pump 22 is connected to the discharge section 33B.
  • blood in the processing space 35 is sent from the discharge section 33B toward the living body LB via the first discharge piping system 12 by driving the second pump 22.
  • the linear member F may have a structure including a holding portion 220 that holds the cells 115 and allows at least some components of blood to pass therethrough.
  • the holding portion 220 in this configuration is formed into a linear shape using, for example, alginic acid having the above-mentioned macroporous structure.
  • the active ingredient-containing solution may be, for example, an infusion solution containing water, electrolytes, nutrients, etc., or an antibody production solution.
  • the chamber has at least an introduction part into which the component in the active ingredient-containing solution is introduced, and a discharge part from which at least the component in the active ingredient-containing solution is discharged, Of the introduction part and the discharge part, at least the discharge part is provided with a filter for keeping the linear member inside the chamber.
  • the active ingredient-containing solution processing device according to Supplementary Note 1. (Additional note 3)
  • the holding part is a mixture containing a polysaccharide and a block copolymer of a polymer having a lower critical solution temperature and a hydrophilic polymer.
  • the specific substance is a virus
  • the affinity cells are angiotensin converting enzyme 2 (ACE2) expressing cells
  • ACE2 angiotensin converting enzyme 2
  • the physiologically active substance is albumin
  • the exocrine cell is a hepatocyte
  • the physiologically active substance is an enzyme
  • the exocrine cell is an enzyme-producing cell
  • the linear member is a core portion filled with a plurality of the cells; A shell part that covers the outer periphery of the core part as the holding part; is a cell fiber having The active ingredient-containing solution processing device according to any one of Supplementary Notes 1 to 7.
  • An active ingredient-containing solution processing device according to any one of Supplementary Notes 1 to 8;
  • An extracorporeal active ingredient-containing solution circulation system comprising: a pump that circulates at least the components of the active ingredient-containing solution from an inlet to an outlet of the chamber.
  • the present invention can be applied to an active ingredient-containing solution treatment device and an extracorporeal active ingredient-containing solution circulation system.
  • Blood processing device active ingredient-containing solution processing device

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Abstract

La présente invention a pour but de procurer un dispositif de traitement de solution contenant des principes actifs pouvant réduire la quantité d'une solution contenant des principes actifs à faire circuler de manière extracorporelle, en réduisant la taille du dispositif. La présente invention comprend : une chambre contenant au moins une partie des composants du sang ; et une fibre cellulaire située à l'intérieur de la chambre et présentant une partie centrale remplie de cellules multiples et une partie extérieure recouvrant la circonférence externe de la partie centrale. Les cellules comprennent : des cellules affinitaires présentant une affinité avec une substance spécifique contenue dans un composant du sang ; et/ou des cellules sécrétoires externes libérant une substance bioactive dans le sang sous la forme d'une sécrétion externe.
PCT/JP2023/016454 2022-04-27 2023-04-26 Dispositif de traitement à l'aide une solution contenant des principes actifs et système de circulation extracorporelle d'une solution contenant des principes actifs WO2023210691A1 (fr)

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JP2022073502 2022-04-27

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5981211A (en) * 1988-05-23 1999-11-09 Regents Of The University Of Minnesota Maintaining cells for an extended time by entrapment in a contracted matrix
WO2015178427A1 (fr) * 2014-05-20 2015-11-26 国立大学法人 東京大学 Microfibre creuse
WO2020032221A1 (fr) * 2018-08-10 2020-02-13 持田製薬株式会社 Microfibre creuse d'alginate
CN112375737A (zh) * 2020-11-12 2021-02-19 常州市第一人民医院 一种生物吸附材料及制备方法、病毒吸附系统和辅助透析循环系统
WO2023085441A1 (fr) * 2021-11-10 2023-05-19 国立大学法人東京大学 Structure macroporeuse

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5981211A (en) * 1988-05-23 1999-11-09 Regents Of The University Of Minnesota Maintaining cells for an extended time by entrapment in a contracted matrix
WO2015178427A1 (fr) * 2014-05-20 2015-11-26 国立大学法人 東京大学 Microfibre creuse
WO2020032221A1 (fr) * 2018-08-10 2020-02-13 持田製薬株式会社 Microfibre creuse d'alginate
CN112375737A (zh) * 2020-11-12 2021-02-19 常州市第一人民医院 一种生物吸附材料及制备方法、病毒吸附系统和辅助透析循环系统
WO2023085441A1 (fr) * 2021-11-10 2023-05-19 国立大学法人東京大学 Structure macroporeuse

Non-Patent Citations (3)

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Title
ANDREI MARIA; STANESCU PAUL O.; DRAGHICI CONSTANTIN; TEODORESCU MIRCEA: "Degradable thermosensitive injectable hydrogels with two-phase composite structure from aqueous solutions of poly(N-isopropylacrylamide-co-5,6-benzo-2-methylene-1,3-dioxepane)—poly(ethylene glycol) triblock copolymers and biopolymers", COLLOID & POLYMER SCIENCE, SPRINGER VERLAG, HEIDELBERG, DE, vol. 295, no. 10, 21 July 2017 (2017-07-21), DE , pages 1805 - 1816, XP036321074, ISSN: 0303-402X, DOI: 10.1007/s00396-017-4161-2 *
MAZARI-ARRIGHI ELSA, OKITSU TERU, TERAMAE HIROKI, AOYAGI HOSHIMI, KIYOSAWA MAHIRO, YANO MARIKO, CHATELAIN FRANÇOIS, FUCHS ALEXANDR: "In vitro proliferation and long-term preservation of functional primary rat hepatocytes in cell fibers", SCIENTIFIC REPORTS, vol. 12, no. 1, XP093105228, DOI: 10.1038/s41598-022-12679-3 *
NAGATA SHOGO, OZAWA FUMISATO, NIE MINGHAO, TAKEUCHI SHOJI: "3D culture of functional human iPSC-derived hepatocytes using a core-shell microfiber", PLOS ONE, vol. 15, no. 6, pages e0234441, XP093105227, DOI: 10.1371/journal.pone.0234441 *

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