Biocompatible device

Abstract

The present invention is a biocompatible device comprising an internal space that can include cells or tissues that secrete a biologically active substance, wherein the internal space is formed by disposing a biocompatible composite membrane including a semi-permeable membrane layer and a non-woven fabric layer in the device such that the semi-permeable membrane layer is on the outer side and the non-woven fabric layer is on the internal-space side, the semi-permeable membrane layer contains a cellulose derivative laminated on a non-woven fabric using the non-woven fabric as a support, and the non-woven fabric layer contains a thermoplastic non-woven fabric.

Description

Biocompatible Devices

[0001] The present invention relates to a biocompatible device for transplanting cells or the like into a living body, a method for producing the same, and a kit for biological transplantation.

[0002] One of the regenerative medical treatments for dysfunctional organs involves transplanting cells or tissues into living organisms, and devices for use in such treatments have been developed. For example, bioartificial pancreases (BAPs), also known as bioartificial islets, have been studied as a treatment for type 1 diabetes. These devices encapsulate pancreatic islets in a polymer gel or semipermeable membrane that can isolate them from the recipient's immune cells and allow permeation of nutrients, insulin, and other nutrients. Various transplantation devices have been reported (Patent Documents 1 to 4). Biocompatible transplantation devices for use in treatments other than type 1 diabetes have also been reported (Patent Documents 5 to 7). An immunoisolation device using a membrane made of polypropylene fiber and an ethylene-vinyl alcohol copolymer has also been reported (Patent Document 8). A transplantation device using chemically crosslinked alginate has also been reported (Patent Documents 9 and 10).

[0003] Japanese Patent Publication No. 2022-502458 Japanese Patent Application Laid-Open No. 2019-097442 International Publication No. 2016 / 184872 Pamphlet Japanese Patent Application Laid-Open No. 2017-502008 Japanese Patent Application Laid-Open No. 2018-515582 Japanese Patent Application Laid-Open No. 2014-200199 Japanese Patent Application Laid-Open No. 2004-209228 International Publication No. 2022 / 092198 Pamphlet International Publication No. 2020 / 262642 Pamphlet International Publication No. 2022 / 137345 Pamphlet

[0004] In the circumstances described above, there has been a demand for new, practical biocompatible devices.

[0005] The present invention has the following features (1) to (5). (1) It is possible to prepare a biocompatible composite membrane including a semipermeable membrane layer containing a cellulose derivative laminated on a nonwoven fabric support, and a nonwoven fabric layer containing a thermoplastic nonwoven fabric. (2) The use of this composite membrane makes it possible to improve the strength of the membrane when covering a hydrogel that can contain cells, tissues, etc. that secrete biologically active substances. (3) The nonwoven fabric surface of the composite membrane can be fused, making it possible to easily fabricate devices by heat sealing. (4) When a device fabricated using this composite membrane is transplanted, it is expected to improve the adhesion rate in vivo and the healing rate of the recipient. (5) By placing the nonwoven fabric surface of the composite membrane inside a device, the hydrogel contained in the internal space of the device, which can contain cells, tissues, etc. that secrete biologically active substances, is immobilized on the nonwoven fabric surface of the composite membrane, making it possible to prevent a decrease in the viability of the cells, etc.

[0006] More specifically, the present invention relates to a biocompatible device having an internal space that can contain cells or tissues that secrete a biologically active substance, the internal space being formed by placing a biocompatible composite membrane comprising a semipermeable membrane layer and a nonwoven fabric layer on the device, with the semipermeable membrane layer on the outside and the nonwoven fabric layer on the internal space side, the semipermeable membrane layer comprising a cellulose derivative laminated on the nonwoven fabric using the nonwoven fabric as a support, and the nonwoven fabric layer comprising a nonwoven fabric having thermoplastic properties, as well as a method for producing the biocompatible device.

[0007] Exemplary aspects of the present invention are as follows [1-1] to [4A-3].

[0008] [1-1] A biocompatible device in which an internal space is formed by a biocompatible composite membrane including a semipermeable membrane layer and a nonwoven fabric layer, wherein the semipermeable membrane layer includes a cellulose derivative formed using a nonwoven fabric as a support, and the nonwoven fabric layer includes the nonwoven fabric having thermoplastic properties, wherein the semipermeable membrane layer is disposed on the outside and the nonwoven fabric layer is disposed on the internal space side of the device, and wherein the internal space contains cells or tissues that secrete a biologically active substance.

[0009] [1-2] The device according to [1-1], which has immunoisolation ability. [1-3] The device according to [1-1] or [1-2], wherein the cellulose derivative is cellulose acetate. [1-4] The device according to any one of [1-1] to [1-3], wherein the nonwoven fabric contains one or more resins selected from the group consisting of polyethylene, polypropylene, and polyethylene terephthalate. [1-5] The device according to any one of [1-1] to [1-4], wherein the internal space is formed by partial fusion of the nonwoven fabric layers.

[0010] [1-6] The device according to any one of [1-1] to [1-5], wherein the composite membrane has a structure in which the cellulose derivative partially penetrates a nonwoven fabric. [1-7] The device according to any one of [1-1] to [1-6], wherein the cells or tissues are encapsulated in an internal space using a hydrogel as a carrier. [1-8] The device according to any one of [1-1] to [1-7], wherein the cells are insulin-secreting cells or pancreatic islets. [1-9] The device according to any one of [1-1] to [1-8], wherein the thickness of the composite membrane is 10 to 1500 μm.

[0011] [1-10] The device according to any one of [1-1] to [1-9], wherein the strength of the composite membrane is 1 MPa or more as measured by a tensile test in accordance with JIS K 7127. [1-11] The device according to any one of [1-1] to [1-10], which is used for biological transplantation. [1-12] A biocompatible composite membrane for producing the device according to any one of [1-1] to [1-11].

[0012] [1-13] A biological transplant kit comprising: a biocompatible device having an internal space formed by a biocompatible composite membrane including a semipermeable membrane layer and a nonwoven fabric layer; and a container containing cells or tissues that secrete a bioactive substance, wherein the cells or tissues are encapsulated in the internal space. [1-14] A method for producing a biocompatible device, comprising the step of covering a hydrogel having cells or tissues that secrete a bioactive substance embedded therein with a biocompatible composite membrane including a semipermeable membrane layer and a nonwoven fabric layer, wherein the semipermeable membrane layer is disposed on the outer side and the nonwoven fabric layer is disposed on the hydrogel side. [1-15] A method for producing a biocompatible device, comprising the step of injecting a solution containing cells or tissues that secrete a bioactive substance mixed in a hydrogelable solution into the internal space formed by the biocompatible composite membrane including a semipermeable membrane layer and a nonwoven fabric layer, and gelling the hydrogelable solution, wherein the semipermeable membrane layer is disposed on the outer side and the nonwoven fabric layer is disposed on the internal space side of the composite membrane.

[0013] [1A-1-1] A biocompatible device having an internal space capable of containing cells or tissues that secrete a biologically active substance, wherein the internal space is formed by placing a biocompatible composite membrane comprising a semipermeable membrane layer and a nonwoven fabric layer on the device, with the semipermeable membrane layer on the outer side and the nonwoven fabric layer on the internal space side, wherein the semipermeable membrane layer comprises a cellulose derivative laminated on the nonwoven fabric using the nonwoven fabric as a support, and the nonwoven fabric layer comprises a nonwoven fabric having thermoplastic properties.

[0014] [1A-1-2] A biocompatible device having an internal space capable of containing cells or tissues that secrete a biologically active substance, wherein the internal space is formed by placing a biocompatible composite membrane consisting of a semipermeable membrane layer and a nonwoven fabric layer on the device, with the semipermeable membrane layer on the outside and the nonwoven fabric layer on the internal space side, wherein the semipermeable membrane layer contains a cellulose derivative laminated on the nonwoven fabric using the nonwoven fabric as a support, and the nonwoven fabric layer contains a nonwoven fabric having thermoplastic properties.

[0015] [1A-1-3] A biocompatible device having an internal space capable of containing cells or tissues that secrete a biologically active substance, wherein the internal space is formed by placing a biocompatible composite membrane having a cellulose derivative laminated on one side of a nonwoven fabric on the device with the surface of the composite membrane on which the cellulose derivative is laminated facing outward.

[0016] [1A-2] The device according to any one of [1A-1-1] to [1A-1-3] above, which has immunoisolation ability. [1A-3] The device according to any one of [1A-1-1] to [1A-2] above, wherein the cellulose derivative is cellulose acetate.

[0017] [1A-4] The device according to any one of [1A-1-1] to [1A-3], wherein the nonwoven fabric comprises one or more resins selected from the group consisting of polyethylene (PE), polypropylene (PP), and polyethylene terephthalate (PET). [1A-5] The device according to any one of [1A-1-1] to [1A-4], wherein the internal space is formed by partial fusion of a surface of the nonwoven fabric in the composite membrane that is not laminated with a cellulose derivative. [1A-6] The device according to any one of [1A-1-1] to [1A-5], wherein the composite membrane has a structure in which the cellulose derivative partially penetrates the nonwoven fabric. [1A-7] The biocompatible device according to any one of [1A-1-1] to [1A-6], wherein the cells secreting the bioactive substance are insulin-secreting cells or pancreatic islets.

[0018] [1A-8] A device described in any one of [1A-1-1] to [1A-7], wherein the cells or tissues secreting the biologically active substance are encapsulated in an internal space using a hydrogel as a carrier.

[0019] [1A-9] The biocompatible device according to [1A-8], wherein the hydrogel is a gel of one or more selected from the group consisting of alginate, collagen, hyaluronan, gelatin, fibronectin, elastin, tenascin, laminin, vitronectin, polypeptide, heparan sulfate, chondroitin, chondroitin sulfate, keratan, keratan sulfate, dermatan sulfate, carrageenan, heparin, chitin, chitosan, agarose, agar, cellulose, methylcellulose, carboxymethylcellulose, glycogen and derivatives thereof, fibrin, fibrinogen, thrombin, polyglutamic acid, polylactic acid, polyglycolic acid, lactic acid-glycolic acid copolymer, gellan gum, xanthan gum, galactomannan, guar gum, locust bean gum, and tara gum.

[0020] [1A-9-1] The biocompatible device according to [1A-8], wherein the hydrogel is a gel of one or more selected from the group consisting of alginate, collagen, hyaluronan, gelatin, fibronectin, elastin, tenascin, laminin, vitronectin, polypeptide, heparan sulfate, chondroitin, chondroitin sulfate, keratan, keratan sulfate, dermatan sulfate, carrageenan, heparin, chitin, chitosan, agarose, agar, cellulose, methylcellulose, carboxymethylcellulose, glycogen, and derivatives thereof.

[0021] [1A-9-2] The biocompatible device according to [1A-8], wherein the hydrogel is a hydrogel obtained by gelling alginic acid or a hydrogel (crosslinked alginic acid gel) obtained by gelling by forming crosslinks using alginic acid derivatives represented by the following formulas (IA) and (II-A): [Alginic acid derivative represented by formula (IA)] Formula (IA): [In formula (IA), (ALG) represents alginic acid; -NHCO- represents an amide bond via any carboxyl group of alginic acid; -L １Ａ - is the following partial structural formula: (wherein x=1 to 50; and where -CH ２ - stands for -C(=O)-, -O-, -NH-, -N(C１－３ alkyl group)-, -S-, C ３～８ It may be substituted with 1 to 15 groups such as a cycloalkyl ring, a benzene ring, a 5- or 6-membered aromatic heterocyclic ring, or a 5- or 6-membered non-aromatic heterocyclic ring; ２ The hydrogen atom of - is a halogen atom, a hydroxyl group, an amino group, C １－３ Alkyl group, —O—C １－３ Alkyl group, —NH(C １－３ alkyl group), -N(C １－３ alkyl group) ２ , -COO-M (M = Na, K, 1 / 2 Ca, hydrogen atom, C １－３ alkyl group), hydroxy C １－３ Alkyl group, C ２－４ Alkanoyl group, —S—C １－３ Alkyl group, —SO ２ -C １－３ A chemically modified alginic acid derivative represented by the formula (wherein the formula does not include the areas outside the dashed lines at both ends):

[0022] [Alginic acid derivative represented by formula (II-A)] Alginic acid derivative represented by the following formula (II-A): [In formula (II-A), (ALG) represents alginic acid; -NHCO- represents an amide bond via any carboxyl group of alginic acid; -L ２Ａ - is the following partial structural formula: wherein y=1 to 50; ２ - stands for -C(=O)-, -O-, -NH-, -N(C １－３ alkyl group)-, -S-, C ３～８ It may be substituted with 1 to 15 groups such as a cycloalkyl ring, a benzene ring, a 5- or 6-membered aromatic heterocyclic ring, or a 5- or 6-membered non-aromatic heterocyclic ring; ２ The hydrogen atom of - is a halogen atom, a hydroxyl group, an amino group, C １－３ Alkyl group, —O—C １－３ Alkyl group, —NH(C １－３ alkyl group), -N(C １－３ alkyl group) ２, -COO-M (M = Na, K, 1 / 2 Ca, hydrogen atom, C １－３ alkyl group), hydroxy C １－３ Alkyl group, C ２－４ Alkanoyl group, —S—C １－３ Alkyl group, —SO ２ -C １－３ A chemically modified alginic acid derivative represented by the formula (which may be substituted with 1 to 10 groups such as alkyl groups, phenyl groups, benzyl groups, 5- to 6-membered aromatic heterocycles, or 5- to 6-membered non-aromatic heterocycles) (the formula does not include the areas outside the dashed lines at both ends).

[0023] [1A-9-3] In the above [1A-9-2], in the chemically modified alginic acid derivative represented by formula (IA), -L １Ａ - is preferably x=2 to 45, and -L １Ａ -CH in - ２ - stands for -C(=O)-, -O-, -NH-, -N(C １－３ may be substituted with 1 to 15 groups such as alkyl groups, cyclohexane rings, 6-membered aromatic heterocycles, 6-membered non-aromatic heterocycles, and benzene rings; -L １Ａ -CH in - ２ The hydrogen atom of - is a hydroxyl group, an amino group, C １－３ Alkyl group, —O—C １－３ Alkyl group, —NH(C １－３ alkyl group), -N(C １－３ alkyl group) ２ , -COO-M (M = Na, K, 1 / 2 Ca, hydrogen atom, C １－３ More preferably, x is 2 to 45, and -L １Ａ -CH in - ２ - stands for -C(=O)-, -O-, -NH-, -N(C １－３ It may be substituted with 1 to 15 groups such as alkyl groups, cyclohexane rings, benzene rings, etc.; more preferably, x=2 to 45, and -L １Ａ -CH in - ２ - may be substituted with 1 to 15 groups such as -C(=O)-, -O-, -NH-, or a benzene ring; particularly preferably, x=3 to 25, and -L １Ａ -CH in - ２- may be substituted with 1 to 15 groups such as -C(=O)-, -O-, -NH-, or a benzene ring; most preferably, x=3 to 15, and -L １Ａ -CH in - ２ - is a linker which may be substituted with 1 to 10 groups such as -C(=O)-, -O-, -NH-, or a benzene ring.

[0024] [1A-9-4] In the above [1A-9-2], -L of the chemically modified alginic acid derivative represented by formula (IA) １Ａ is preferably as shown in the table below: and more preferably, a linker selected from the following table: A linker (wherein the formula does not include the areas outside the dashed lines at both ends) selected from the following: A linker (wherein the formula does not include the areas outside the dashed lines at both ends) selected from the following table is particularly preferred: (in each formula, the areas outside the dashed lines at both ends are not included).

[0025] [1A-9-5] In the above [1A-9-2], Aky of the alginic acid derivative represented by formula (IA) is preferably a 7- to 9-membered cyclic alkyne group (-CH ２ The hydrogen atom of - is a halogen atom, a hydroxyl group, an amino group, a keto group, C １－３ Alkyl group, —O—C １－３ Alkyl group, —NH(C １－３ alkyl group), -N(C １－３ alkyl group) ２ , -COO-M (M = Na, K, 1 / 2 Ca, hydrogen atom, C １－３ The cyclic alkyne group may be substituted with 1 to 5 groups selected from groups such as C ３～８ and more preferably an 8-membered cyclic alkyne group (cyclooctyne group) (the —CH of the cyclic alkyne group is optionally condensed with 1 to 3 cycloalkyl rings, benzene rings, or 5- or 6-membered aromatic heterocycles). ２ The hydrogen atom of - is a halogen atom, a keto group, C１－３ Alkyl group, —O—C １－３ and the cyclic alkyne group may be fused with 1 to 3 cyclopropane rings, benzene rings, or 5-membered aromatic heterocycles; more preferably, the cyclic alkyne group is a group represented by the following partial structural formula: (wherein the formula does not include the areas to the right of the dashed lines at both ends); and even more preferably, a group selected from the following partial structural formula: (wherein the formula does not include the areas to the right of the dashed lines at both ends) is a cyclic alkyne group selected from the group consisting of: (wherein the right side of the dashed lines at both ends is not included); most preferably, a cyclic alkyne group selected from the group consisting of the following partial structural formula: (wherein the right side of the dashed lines at both ends is not included)

[0026] [1A-9-6] In the above [1A-9-2], in the alginic acid derivative represented by formula (II-A), -L ２Ａ - is preferably y=5 to 40, and -L ２Ａ -CH in - ２ - stands for -C(=O)-, -O-, -NH-, -N(C １－３ may be substituted with 1 to 15 groups such as alkyl group, cyclohexane ring, 6-membered aromatic heterocyclic ring, 6-membered non-aromatic heterocyclic ring, benzene ring, etc., ２Ａ -CH in - ２ The hydrogen atom of - is a hydroxyl group, an amino group, C １－３ Alkyl group, —O—C １－３ Alkyl group, —NH(C １－３ alkyl group), -N(C １－３ alkyl group) ２ , -COO-M (M = Na, K, 1 / 2 Ca, hydrogen atom, C １－３ More preferably, y is 5 to 40, and -L ２Ａ -CH in - ２ - stands for -C(=O)-, -O-, -NH-, -N(C １－３It is more preferable that y is 5 to 40, and -L ２Ａ -CH in - ２ - stands for -C(=O)-, -O-, -NH-, -N(C １－３ It is particularly preferred that y is 5 to 20, and -L ２Ａ -CH in - ２ - may be substituted with 1 to 10 groups such as -C(=O)-, -O-, -NH-, or a benzene ring; most preferably, y=5 to 15, and -L ２Ａ -CH in - ２ - is a linker which may be substituted with 1 to 10 groups such as -C(=O)-, -O-, -NH-, or a benzene ring.

[0027] [1A-9-7] In the above [1A-9-2], -L of the alginic acid derivative represented by formula (II-A) ２Ａ is preferably as shown in the table below: and more preferably a linker selected from the following: A linker (wherein the formula does not include the areas outside the dashed lines at both ends) selected from the following: A linker (wherein the formula does not include the areas outside the dashed lines at both ends) selected from the following table is particularly preferred: is a linker selected from the following (wherein the formula does not include the areas outside the dashed lines at both ends):

[0028] [1A-10] In the above [1A-9-2], the alginic acid derivative represented by formula (IA) is preferably represented by the following formulae (A01) to (A15): [In each formula, (ALG) represents alginic acid; and -NHCO- bonded to (ALG) represents an amide bond via any carboxyl group of alginic acid]; More preferably, it is selected from the following formula (A01) or (A02): In each formula, (ALG) represents alginic acid; and -NHCO- bonded to (ALG) represents an amide bond via any carboxyl group of alginic acid.

[0029] [1A-11] In the above [1A-9-2], the alginic acid derivative represented by formula (II-A) is preferably represented by the following formulas (N01) to (N15): [In each formula, (ALG) represents alginic acid; and -NHCO- bonded to (ALG) represents an amide bond via any carboxyl group of alginic acid]; More preferably, the following formulae (N01) to (N04): In each formula, (ALG) represents alginic acid; and -NHCO- bonded to (ALG) represents an amide bond via any carboxyl group of alginic acid.

[0030] [1A-12] The biocompatible device according to any one of [1A-1-1] to [1A-11], wherein the thickness of the composite film is about 10 to about 1500 μm. [1A-13] The biocompatible device according to any one of [1A-1-1] to [1A-12], wherein the strength of the composite film measured by a tensile test in accordance with JIS K 7127 is about 1 MPa or more.

[0031] [1A-14] The biocompatible device according to any one of [1A-1-1] to [1A-13] above, which is used for biological transplantation. [1A-15] The biocompatible device according to any one of [1A-1-1] to [1A-14] above, which is formed by laminating a cellulose acetate derivative on one side of a nonwoven fabric by the Robb-Srirajan method. [1A-16] A biocompatible composite membrane for producing the biocompatible device according to any one of [1A-1-1] to [1A-15] above.

[0032] [1A-17] A biological transplant kit comprising a biocompatible device in which an internal space is formed by a biocompatible composite membrane in which a cellulose derivative is laminated on one side of a nonwoven fabric, and a carrier containing cells or tissues that secrete a biologically active substance, the cells or tissues being used so as to be enclosed in the internal space.

[0033] [1A-18-1] A method for producing a biocompatible device, comprising the step of covering a hydrogel, into which cells or tissues that secrete a bioactive substance are embedded, with a biocompatible composite membrane including a semipermeable membrane layer and a nonwoven fabric layer, wherein the semipermeable membrane layer is disposed on the outer side and the nonwoven fabric layer is disposed on the hydrogel side. [1A-18-2] A method for producing a biocompatible device, comprising the step of covering a hydrogel, into which cells or tissues that secrete a bioactive substance are embedded, with a biocompatible composite membrane comprising a nonwoven fabric on one side of which a cellulose derivative is laminated, wherein the side of the biocompatible composite membrane onto which the cellulose derivative is laminated is disposed on the outer side. [1A-18-3] The method for producing a biocompatible device according to [1A-18-1] or [1A-18-2], wherein the hydrogel is a hydrogel into which one or more of the gellable substances described in any one of [1A-9] to [1A-11] has been gelled.

[0034] [1A-19-1] A method for manufacturing a biocompatible device, comprising the steps of: injecting a solution containing cells or tissues that secrete a bioactive substance, the solution being mixed with a hydrogelable solution, into an internal space formed by a biocompatible composite membrane comprising a semipermeable membrane layer and a nonwoven fabric layer, and gelling the hydrogelable solution, wherein in the step, the semipermeable membrane layer of the composite membrane faces the external side and the nonwoven fabric layer faces the internal space. [1A-19-2] A method for manufacturing a biocompatible device, comprising the steps of injecting a mixed solution containing cells or tissues that secrete a bioactive substance, the solution being mixed with a hydrogelable solution, into an internal space formed by a biocompatible composite membrane having a cellulose derivative laminated on one side of a nonwoven fabric, and gelling the mixed solution, wherein in the step, the surface of the biocompatible composite membrane on which the cellulose derivative is laminated faces the external side. [1A-19-3] The method for producing a biocompatible device according to [1A-19-1] or [1A-19-2], wherein the hydrogelable solution is a solution containing one or more of the gellable substances described in any one of [1A-9] to [1A-11].

[0035] [2-1] A biocompatible composite membrane comprising a semipermeable membrane layer and a nonwoven fabric layer, wherein the semipermeable membrane layer comprises a cellulose derivative formed using a nonwoven fabric as a support, and the nonwoven fabric layer comprises the nonwoven fabric having thermoplastic properties, and is used to encapsulate cells or tissues. [2-2] The composite membrane according to [2-1], which has immunoisolation ability. [2-3] The composite membrane according to [2-1] or [2-2], wherein the cellulose derivative is cellulose acetate. [2-4] The composite membrane according to any one of [2-1] to [2-3], wherein the nonwoven fabric comprises one or more resins selected from the group consisting of polyethylene, polypropylene, and polyethylene terephthalate. [2-5] The composite membrane according to any one of [2-1] to [2-4], wherein an internal space can be formed by partial fusion of the nonwoven fabric layers. [2-6] The composite membrane according to any one of [2-1] to [2-5], wherein the composite membrane has a structure in which the cellulose derivative partially penetrates the nonwoven fabric. [2-7] The composite membrane according to any one of [2-1] to [2-6], wherein the cells or tissues are encapsulated in an internal space using a hydrogel as a carrier. [2-8] The composite membrane according to any one of [2-1] to [2-7], wherein the cells or tissues are cells or tissues that secrete a biologically active substance. [2-9] The composite membrane according to any one of [2-1] to [2-8], wherein the cells are insulin-secreting cells or pancreatic islets. [2-10] The composite membrane according to any one of [2-1] to [2-9], wherein the thickness is 10 to 1500 μm. [2-11] The composite membrane according to any one of [2-1] to [2-10], wherein the strength of the composite membrane measured by a tensile test in accordance with JIS K 7127 is 1 MPa or more. [2-12] The composite membrane according to any one of [2-1] to [2-11], wherein the composite membrane is used for producing a biotransplant device.

[0036] [2A-1-1] A biocompatible composite membrane comprising a semipermeable membrane layer and a nonwoven fabric layer, wherein the semipermeable membrane layer comprises a cellulose derivative laminated on the nonwoven fabric using the nonwoven fabric as a support, and the nonwoven fabric layer comprises a thermoplastic nonwoven fabric, and is used for encapsulating cells or tissues. [2A-1-2] A biocompatible composite membrane comprising a semipermeable membrane layer and a nonwoven fabric layer, wherein the semipermeable membrane layer comprises a cellulose derivative laminated on the nonwoven fabric using the nonwoven fabric as a support, and the nonwoven fabric layer comprises a thermoplastic nonwoven fabric, and is used for encapsulating cells or tissues. [2A-1-3] A biocompatible composite membrane comprising a cellulose derivative laminated on one side of a nonwoven fabric, and is used for encapsulating cells or tissues.

[0037] [2A-2] The composite membrane according to any one of [2A-1-1] to [2A-1-3] above, which has immunoisolation ability. [2A-3] The composite membrane according to any one of [2A-1-1] to [2A-2] above, wherein the cellulose derivative is cellulose acetate. [2A-4] The composite membrane according to any one of [2A-1-1] to [2A-3] above, wherein the nonwoven fabric comprises one or more resins selected from the group consisting of polyethylene (PE), polypropylene (PP), and polyethylene terephthalate (PET).

[0038] [2A-5] The composite membrane according to any one of [2A-1-1] to [2A-4], wherein an internal space can be formed by partial fusion of the nonwoven fabric surface of the composite membrane on which the cellulose derivative is not laminated. [2A-6] The composite membrane according to any one of [2A-1-1] to [2A-5], wherein the composite membrane has a structure in which the cellulose derivative partially penetrates the nonwoven fabric. [2A-7] The composite membrane according to any one of [2A-1-1] to [2A-6], wherein cells or tissues are encapsulated using a hydrogel as a carrier in an internal space formed by partial fusion of the nonwoven fabric surface of the composite membrane on which the cellulose derivative is not laminated. [2A-7-1] The composite membrane according to [2A-7], wherein the hydrogel is a hydrogel obtained by gelling one or more of the gellable substances according to any one of [1A-9] to [1A-11].

[0039] [2A-8] The composite membrane according to [2A-7] or [2A-7-1] above, wherein the cells or tissues are cells or tissues that secrete a biologically active substance. [2A-9] The composite membrane according to any one of [2A-1-1] to [2A-8] above, wherein the cells are insulin-secreting cells or pancreatic islets. [2A-10] The composite membrane according to any one of [2A-1-1] to [2A-9] above, wherein the thickness is about 10 to about 1500 μm. [2A-11] The composite membrane according to any one of [2A-1-1] to [2A-10] above, wherein the strength of the membrane measured by a tensile test in accordance with JIS K 7127 is about 1 MPa or more. [2A-12] The composite membrane according to any one of [2A-1-1] to [2A-11] above, wherein the composite membrane is used for producing a bioimplant device. [2A-13] The composite membrane according to any one of [2A-1-1] to [2A-12], wherein the composite membrane is formed by laminating a cellulose acetate derivative on one side of a nonwoven fabric by the Robb-Srirajan method.

[0040] [3-1] The device according to any one of [1-7] to [1-11], or the composite membrane according to any one of [2-7] to [2-12], wherein the hydrogel comprises one or more gelled hydrogels selected from the group consisting of alginate, collagen, hyaluronan, gelatin, fibronectin, elastin, tenascin, laminin, vitronectin, polypeptide, heparan sulfate, chondroitin, chondroitin sulfate, keratan, keratan sulfate, dermatan sulfate, carrageenan, heparin, chitin, chitosan, agarose, agar, cellulose, methylcellulose, carboxymethylcellulose, glycogen and derivatives thereof, fibrin, fibrinogen, thrombin, polyglutamic acid, polylactic acid, polyglycolic acid, lactic acid-glycolic acid copolymer, gellan gum, xanthan gum, galactomannan, guar gum, locust bean gum, and tara gum.

[0041] [3-2] The device according to any one of [1-7] to [1-11] or the composite membrane according to any one of [2-7] to [2-12], wherein the hydrogel comprises a hydrogel gelled with one or more selected from the group consisting of alginate, collagen, hyaluronan, gelatin, fibronectin, elastin, tenascin, laminin, vitronectin, polypeptide, heparan sulfate, chondroitin, chondroitin sulfate, keratan, keratan sulfate, dermatan sulfate, carrageenan, heparin, chitin, chitosan, agarose, agar, cellulose, methylcellulose, carboxymethylcellulose, glycogen, and derivatives thereof.

[0042] [3-3] The device according to any one of [1-7] to [1-11] or the composite membrane according to any one of [2-7] to [2-12], wherein the hydrogel comprises a hydrogel formed by gelling alginic acid or an alginic acid derivative. [3-4] The device according to any one of [1-7] to [1-11] or the composite membrane according to any one of [2-7] to [2-12], wherein the hydrogel comprises a hydrogel formed by gelling an alginic acid derivative by chemical crosslinking.

[0043] [3-5] The device or composite membrane according to [3-4], wherein the alginic acid derivative comprises an alginic acid derivative represented by the following formula (HI) and formula (H-II): [Alginic acid derivative represented by formula (HI)] The alginic acid derivative represented by the following formula (HI): [In formula (HI), (ALG) represents alginic acid; -NHCO- represents an amide bond via any carboxyl group of alginic acid; -L １ - is a divalent linker bonding to a cyclic alkyne group (Akn); [Alginic acid derivative represented by formula (H-II)] Alginic acid derivative represented by the following formula (H-II): [In formula (H-II), (ALG) represents alginic acid; -NHCO- represents an amide bond via any carboxyl group of alginic acid; -L ２ - is a divalent linker that bonds to the azide group.

[0044] [3-6] In formula (HI), -L１ - is - (CH ２ ) ｎ１ - (where n1=1 to 50, and -CH ２ - may be replaced by 1 to 10 groups selected from -CO-, -CONH-, -NHCO-, -O-CONH-, -NHCO-O-, -O- and -NH-, or a benzene ring, ２ The hydrogen atom of - is C １－3 C substituted with alkyl or phenyl groups １－3 Akn is an 8-membered cyclic alkyne group (which may be further fused with one or two benzene rings, cyclopropane rings, or 1,2,3-triazole rings, and the fused rings may have -L １ -, and -CH in the 8-membered cyclic alkyne group ２ - may be an 8-membered ring group substituted by 1 or 2 groups selected from -C(=O)-, -CONH-, and -NH-, and -CH ２ The hydrogen atom of - is C １－３ an alkyl group, a fluorine atom, a hydroxyl group, or C １－３ and wherein in formula (H-II), -L ２ -ga- (CH ２ ) ｎ２ - (where n2=1 to 50, and -CH ２ - may be replaced by 1 to 10 groups selected from -CONH-, -NHCO-, -O- and -NH-, or a benzene ring or a pyridine ring, and the -CH ２ The hydrogen atom of - is C １－3 The device or composite film according to [3-5], wherein the alkyl group is a substituted or unsubstituted alkyl group.

[0045] [3-7] In formula (HI), -L １ - is - (CH ２ ) ｎ１ - (where n1=2 to 15, and -CH ２- may be replaced by 1 to 5 groups selected from -CO-, -CONH-, -NHCO-, -O-CONH-, -NHCO-O-, -O- and -NH-, or a benzene ring, ２ The hydrogen atom of - is C １－3 C substituted with alkyl or phenyl groups １－3 Akn is an 8-membered cyclic alkyne group (which may be further fused with 1 or 2 benzene rings, and the 8-membered cyclic alkyne group may be fused with -CH ２ In the formula (H-II), -L may be an 8-membered ring group in which - is replaced by -NH-. ２ -ga- (CH ２ ) ｎ２ - (where n2=2 to 15, and -CH ２ The device or composite film according to [3-5] or [3-6], wherein - may be replaced by 1 to 5 groups selected from -CONH-, -NHCO-, -O-, and -NH-, or a benzene ring.

[0046] [3-8] The device or composite membrane according to any one of [3-4] to [3-7], wherein the chemical crosslinking is achieved by combining alginic acid derivatives represented by the following formulas (H-I) and (H-II): [Alginic acid derivative represented by formula (H-I)] The alginic acid derivative represented by the following formula (H-I): [In formula (HI), (ALG) represents alginic acid; -NHCO- represents an amide bond via any carboxyl group of alginic acid; -L １ - represents the following partial structural formula (each formula does not include the areas outside the wavy lines at both ends): Akn represents a divalent linker selected from the group consisting of: wherein the asterisk represents a chiral center]; [Alginic acid derivatives represented by formula (H-II)] The following formula (H-II): [In formula (H-II), (ALG) represents alginic acid; -NHCO- represents an amide bond via any carboxyl group of alginic acid; -L ２ - represents the following partial structural formula (each formula does not include the areas outside the wavy lines at both ends): The alginic acid derivative is represented by the following formula:

[0047] [3-9] The alginic acid derivative represented by formula (H-I) is the following formula (A01) or (A02), and the alginic acid derivative represented by formula (H-II) is the following formula (N01), (N02), (N03) or (N04). [3-5] to [3-8] The device or composite membrane according to any one of the above. [In each formula, (ALG) represents alginic acid; -NHCO- bonded to (ALG) represents an amide bond via any carboxyl group of alginic acid]

[0048] [3-10] The chemically crosslinked alginic acid derivative is such that any carboxyl group of the first alginic acid and any carboxyl group of the second alginic acid are represented by the following formula (H-III-L): [In formula (H-III-L), —CONH— and —NHCO— at both ends represent an amide bond via any carboxyl group of alginic acid; １ - is as defined in any one of [3-5] to [3-8]; -L ２ - is as defined in any one of [3-5] to [3-8]; X is the following partial structural formula: (In each formula, the areas outside the dashed lines at both ends are not included, and an asterisk represents a chiral center.) The device or composite membrane according to any one of [3-4] to [3-9], wherein the alginate is a cross-linked alginate bonded via a cyclic group selected from the group consisting of:

[0049] [3-11] In the formula (H-III-L), -L １ - represents the following partial structural formula (each formula does not include the areas outside the dashed lines at both ends): -L is a bivalent linker selected from the group ２ - represents the following partial structural formula (each formula does not include the areas outside the dashed lines at both ends): X is a divalent linker selected from the group consisting of the following partial structural formulas (in each formula, the areas outside the dashed lines at both ends are not included): The device or composite membrane according to [3-10], wherein the cyclic group is selected from the group consisting of:

[0050] [3-12] In the formula (H-III-L), -L ２ -XL １ The combination of - is represented by the following partial structural formula (each formula does not include the areas outside the dashed lines at both ends): The device or composite membrane according to [3-10] or [3-11], selected from the group consisting of:

[0051] [3-13] In the formula (H-III-L), -L ２ -XL １ The combination of - is represented by the following partial structural formula (the formula does not include the areas outside the dashed lines at both ends): The device or composite membrane according to any one of [3-10] to [3-12], selected from the group consisting of:

[0052] [3A] A hydrogel (crosslinked alginic acid) gelled by forming crosslinks using the alginic acid derivatives represented by the formula (IA) and the formula (II-A) is a hydrogel represented by the following formula (III-Lx): [In formula (III-Lx), —NHCO— on the right end represents an amide bond derived from the alginic acid derivative of formula (IA), and —CONH— on the left end represents an amide bond derived from the alginic acid derivative of formula (II-A); １Ａ - and -L ２Ａ - is the same as defined in the above embodiment [1A-9-2]; -X Ａ - is the following partial structural formula: (In the formula, C ５－９ -CH in the cycloalkene ring ２ - may be substituted with 1 to 4 groups selected from the group consisting of -NH-, -S-, -O-, or =C(=O); C ５－９ -CH in the cycloalkene ring ２ The hydrogen atom of - is a halogen atom, a hydroxyl group, an amino group, a keto group, C １－３ Alkyl group, —O—C １－３ Alkyl group, —NHC １－３alkyl group, —N(C １－３ alkyl group) ２ , -COO-M (M = Na, K, 1 / 2 Ca, hydrogen atom, C １－３ alkyl group), and the like; ５－９ The cycloalkene ring has C ３－８ may be condensed with 1 to 3 cycloalkyl rings, benzene rings, or 5- or 6-membered aromatic heterocycles; ５－９ The cycloalkene ring has C ３－８ When a cycloalkyl ring, a benzene ring, or a 5- or 6-membered aromatic heterocycle is condensed, -L １Ａ - is the C ３－８ The biocompatible device according to [1A-9-2] above is a cross-linked alginate gel bonded via a cyclic group represented by the formula (the formula does not include the areas outside the dashed lines at both ends):

[0053] [3A-1] In the above [3A], preferred, more preferred, further preferred, particularly preferred, most preferred -L １Ａ - is the -L described in [1A-9-3] above. １Ａ [3A-1-2] In the above [3A], -L is preferably, more preferably, further preferably, particularly preferably, １Ａ - is the -L described in [1A-9-4] above. １Ａ - is the same as the definition.

[0054] [3A-2] In the above [3A], preferred, more preferred, further preferred, particularly preferred, most preferred -L ２Ａ - is the -L described in [1A-9-6] above. ２Ａ [3A-2-2] In the above [3A], -L is preferably, more preferably, further preferably, particularly preferably, ２Ａ - is the -L described in [1A-9-7] above. ２Ａ - is the same as the definition.

[0055] [3A-3] In the above aspect [3A], -X Ａ - is preferably the following partial structural formula: (in each formula, the areas outside the dashed lines at both ends are not included); more preferably, a cyclic group selected from the following partial structural formula: (in each formula, the areas outside the dashed lines at both ends are not included); more preferably, a cyclic group selected from the group consisting of the following partial structural formulas: (In each formula, the areas outside the dashed lines at both ends are not included.)

[0056] [3A-4] -L described in [3A] to [3A-3] １Ａ -, -L ２Ａ -, -X Ａ By appropriately combining the definitions of -, a preferred embodiment of the formula (III-Lx) in the crosslinked alginate gel can be arbitrarily formed. [3A-5] In the formula (III-Lx) of the above [3A], -L １Ａ -X Ａ -L ２Ａ Specific preferred combinations of - are shown in the table below: More preferred combinations are those listed in the table below: It is a combination described in

[0057] [4-1] The biological transplant kit according to [1-13], wherein the hydrogel contains the alginic acid derivative or cross-linked alginic acid according to any one of [3-5] to [3-13]. [4-2] The manufacturing method according to [1-14], wherein the hydrogel contains the alginic acid derivative or cross-linked alginic acid according to any one of [3-5] to [3-13]. [4-3] The manufacturing method according to [1-15], wherein the solution contains an alginic acid solution containing the alginic acid derivative or cross-linked alginic acid according to any one of [3-5] to [3-13].

[0058] [4A-1] The biological transplant kit described in [1-13] or [1A-17] above, wherein the hydrogel comprises the alginic acid derivative or crosslinked alginic acid described in any one of [1A-9] to [1A-11] or [3A] to [3A-5] above. [4A-2] The manufacturing method described in [1-14], [1A-18-1] or [1A-18-2] above, wherein the hydrogel comprises the alginic acid derivative or crosslinked alginic acid described in any one of [1A-9] to [1A-11] or [3A] to [3A-5] above. [4A-3] The manufacturing method described in [1-15], [1A-19-1] or [1A-19-2] above, wherein the solution comprises an alginic acid solution containing the alginic acid derivative or crosslinked alginic acid described in any one of [1A-9] to [1A-11] or [3A] to [3A-5] above.

[0059] One aspect of the present invention provides a novel biocompatible device that can be used in practice. Another aspect of the present invention provides a biocompatible composite membrane that can be used as a component of the device. Furthermore, another aspect of the present invention provides a method for producing the device and a kit for biological transplantation.

[0060] Figure 1 is a schematic diagram of a biocompatible device according to one embodiment of the present invention. Figure 2 is a schematic diagram of a cross section of a biocompatible composite membrane according to one embodiment of the present invention. Figure 3 is a schematic diagram of a cross section of a biocompatible device according to one embodiment of the present invention. Figure 4 is a photograph of a biocompatible device according to one embodiment of the present invention.

[0061] Regarding the numerical ranges described herein, the upper and lower limits can be arbitrarily combined. For example, when a numerical range is described as "3.0 to 10, 3.2 to 8," the ranges "3.0 to 8" and "3.2 to 10" are also included in the numerical ranges described herein. Furthermore, when a numerical range is described as "3.0 or more, 3.2 or more, and 10 or less, 8 or less," the ranges "3.0 to 8" and "3.2 to 10" are also included in the numerical ranges described herein. Furthermore, as a numerical range described herein, for example, "5 to 10" means a range of "5 or more, 10 or less." Furthermore, in specifying the upper and lower limits described herein, the numerical range from the lower limit to the upper limit can be specified by appropriately selecting from the respective options and combining them arbitrarily. In addition, multiple combinations of the various requirements described as preferred aspects described herein can be used.

[0062] 1. Biocompatible Devices Provided herein is a biocompatible device having an internal space formed by a biocompatible composite membrane including a semipermeable membrane layer and a nonwoven fabric layer, wherein the semipermeable membrane layer contains a cellulose derivative formed using a nonwoven fabric as a support, and the nonwoven fabric layer contains the thermoplastic nonwoven fabric, with the semipermeable membrane layer positioned on the outside and the nonwoven fabric layer facing the internal space, and the internal space containing cells or tissue. Also provided is a biocompatible device having an internal space capable of containing cells or tissue secreting a biologically active substance, wherein the internal space is formed by placing a biocompatible composite membrane including / consisting of a semipermeable membrane layer and a nonwoven fabric layer on the outside and the nonwoven fabric layer facing the internal space, with the semipermeable membrane layer containing a cellulose derivative laminated on the nonwoven fabric using the nonwoven fabric as a support, and the nonwoven fabric layer containing a thermoplastic nonwoven fabric. Furthermore, there is provided a biocompatible device having an internal space capable of containing cells or tissues secreting a biologically active substance, the internal space being formed by placing a biocompatible composite membrane having a cellulose derivative laminated on one side of a nonwoven fabric, with the surface of the composite membrane on which the cellulose derivative is laminated facing outward in the device. As used herein, a "biocompatible device" refers to a device in which an internal space is formed by a biocompatible composite membrane comprising a semipermeable membrane layer and a nonwoven fabric layer, regardless of whether cells or tissues are encapsulated therein. In other words, a device in which an internal space is formed by a biocompatible composite membrane having a cellulose derivative laminated on one side of a nonwoven fabric. In this specification, "biocompatible device" and "biocompatible composite membrane" may be simply referred to as "device" and "composite membrane." In this specification, "biocompatibility" means a property that is considered to pose no problems in terms of biological safety when maintained in a living body, and examples of this include the property of not inducing interactions between the various biological materials that make up the biocompatible device and the living body, or local reactions in tissues adjacent to the biological materials, or systemic reactions, or the like, or of reducing such reactions; possessing these properties is said to be biocompatible.In this specification, the terms "biocompatible device" and "biocompatible composite membrane" mean "device having biocompatibility" and "composite membrane having biocompatibility", respectively.

[0063] A preferred embodiment of the biocompatible device of the present invention includes not only a semipermeable membrane layer but also a nonwoven fabric layer. This device has suitable physical properties for long-term in vivo use, including transplantation into a living organism, while also providing appropriate substance permeability and good flexibility and strength. Furthermore, the internal space of the biocompatible device of the present invention, for example, when cells or tissues are encapsulated using a hydrogel as a carrier, exhibits excellent shape retention of the hydrogel. Therefore, the internal space of the biocompatible device of the preferred embodiment can provide an environment suitable for the survival and functional maintenance of cells or tissues, allowing the encapsulated cells or tissues to exert their functions (e.g., hypoglycemic effect due to glucose-responsive insulin secretion, when insulin-producing cells are used) for a long period of time. Furthermore, by arranging the semipermeable membrane layer and the nonwoven fabric layer (a biocompatible composite membrane in which a cellulose derivative is laminated on one side of a nonwoven fabric) as described above, the biocompatible device of the preferred embodiment of the present invention can be a highly safe device with minimal adhesions, inflammation, etc.

[0064] A preferred embodiment of the device has immunoisolation capability. As used herein, "immunoisolation capability" refers to the property of isolating cells or tissues from the recipient's immune system. By providing this property, immune rejection reactions (e.g., those due to cellular immune responses and humoral immune responses) in the recipient can be prevented upon transplantation. The immunoisolation capability can be adjusted by selecting the permeability of the semipermeable membrane described below so as to block the entry of antibodies, immune cells, and the like.

[0065] As used herein, "crosslinking (reaction)" refers to "ionic crosslinking (reaction)" or "chemical crosslinking (reaction)." As used herein, "ionic crosslinking (reaction)" refers to the formation of an ionic crosslink (ionic bond) between certain alginic acid or alginic acid derivatives via bonding with a divalent or higher metal ion. Furthermore, as used herein, "chemical crosslinking (reaction)" refers to "crosslinking (reaction) by chemical bonding," meaning that a chemical crosslink (chemical bond) is formed between the alginic acid derivatives by performing a chemical reaction (e.g., Huisgen reaction) using a certain alginic acid derivative. Each component of the biocompatible device will be described in detail below.

[0066] 1-1. Biocompatible Composite Membrane In this specification, the term "biocompatible composite membrane" includes a semipermeable membrane layer and a nonwoven fabric layer. The semipermeable membrane layer includes a cellulose derivative formed using a nonwoven fabric as a support, and the nonwoven fabric layer includes the nonwoven fabric having thermoplastic properties. The semipermeable membrane layer includes a cellulose derivative laminated on the nonwoven fabric using the nonwoven fabric as a support, and the nonwoven fabric layer includes a nonwoven fabric having thermoplastic properties. The biocompatible composite membrane is a membrane in which a cellulose derivative is laminated on one side of a nonwoven fabric, and the nonwoven fabric is thermoplastic.

[0067] (Semipermeable membrane layer) The semipermeable membrane layer comprises a cellulose derivative formed using a nonwoven fabric as a support. The semipermeable membrane layer comprises a cellulose derivative laminated on a nonwoven fabric using a nonwoven fabric as a support. In some embodiments, the semipermeable membrane layer comprises a semipermeable membrane containing the cellulose derivative. In other embodiments, the semipermeable membrane layer consists of a semipermeable membrane containing the cellulose derivative. As used herein, a "semipermeable membrane" is a membrane that allows only molecules or ions below a certain size to pass through. A semipermeable membrane is a system of a solute that does not pass through the semipermeable membrane and a solvent that exhibits permeability; when solutions of two concentrations are brought into contact via the semipermeable membrane, osmotic pressure is generated across the membrane, allowing only the solvent to pass through.

[0068] As used herein, the term "cellulose derivatives" is a general term for compounds in which different substituents have been introduced into hydroxy groups contained in cellulose molecules, and are classified into cellulose ethers formed by ether bonds (e.g., methyl cellulose, ethyl cellulose, ethyl methyl cellulose, carboxymethyl cellulose, carboxyethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, benzyl cellulose, trityl cellulose, cyanoethyl cellulose, aminoethyl cellulose, etc.) and cellulose esters formed by ester bonds (e.g., cellulose acetate, diacetyl cellulose, triacetyl cellulose, cellulose propionate, cellulose butyrate, etc.).

[0069] In some embodiments, the cellulose derivative includes a cellulose ester. In some embodiments, the cellulose ester includes cellulose acetate. In other embodiments, the semipermeable membrane layer may contain a resin other than a cellulose derivative as a component of the semipermeable membrane. Such resins are not particularly limited, but examples include polysulfone-based polymers, polyacrylonitrile-based polymers, polyamide-based polymers, and polycarbonate-based polymers. In one embodiment of the present invention, the content of the cellulose derivative in the components of the semipermeable membrane is about 50 to about 100 mass%, about 60 to about 100 mass% in another embodiment, and about 70 to about 100 mass% in yet another embodiment, based on the total amount of the components of the semipermeable membrane. In another embodiment of the present invention, the content of the resin other than a cellulose derivative in the components of the semipermeable membrane is about 1 to about 50 mass%, about 1 to about 40 mass% in another embodiment, and about 1 to about 30 mass% in yet another embodiment, based on the total amount of the components of the semipermeable membrane.

[0070] In this specification, unless a preferred range is stated, the term "about" may include values ​​up to ±10% of the numerical value, and in some embodiments, up to ±20% of the numerical value.

[0071] In another embodiment of the present invention, the semipermeable membrane is made of a cellulose derivative. That is, in this embodiment, the semipermeable membrane is substantially free of resins other than cellulose derivatives. As used herein, "substantially free" does not exclude cases where a certain component is inevitably mixed in during the manufacturing process, but it is preferable that such unavoidable components be kept to a minimum. In one embodiment of the present invention, in which the semipermeable membrane is substantially free of resins other than cellulose derivatives, the content of resins other than cellulose derivatives is, specifically, less than about 0.1% by mass, less than about 0.05% by mass in another embodiment, and less than about 0.01% by mass in yet another embodiment, based on the total amount of the constituent components of the semipermeable membrane. Furthermore, the semipermeable membrane of one embodiment of the present invention is substantially free of ethylene-vinyl alcohol-based copolymers. The content of ethylene-vinyl alcohol-based copolymers in the constituent components of the semipermeable membrane is, specifically, less than about 0.1% by mass, less than about 0.05% by mass in another embodiment, and less than about 0.01% by mass in yet another embodiment, based on the total amount of the constituent components of the semipermeable membrane.

[0072] Examples of semipermeable membranes that form the semipermeable membrane layer include those having properties and functions equivalent to membranes or tubes used in dialysis, and examples of trade names include Cellu-Sep T Tubular Membrane (Membrane Filtration Products), Spectra Biotec Membrane (REPLIGEN), Spectra / Por CE Dialysis Tubing (REPLIGEN), etc. The semipermeable membrane can be produced, for example, by dissolving a resin containing a cellulose derivative in a solvent and solidifying the dissolved resin.

[0073] The semipermeable membrane used herein has a "molecular weight cutoff." The "molecular weight cutoff" refers to the maximum molecular weight that is not substantially blocked. Molecules with a molecular weight above the molecular weight cutoff are substantially prevented from passing through the semipermeable membrane. In some embodiments, the "molecular weight cutoff" of the semipermeable membrane is 100 kDa, and in another embodiment, 300 kDa. By using a semipermeable membrane with a molecular weight cutoff value within the above range, a semipermeable membrane layer can be formed that maintains a constant rate of permeation of low molecular weight components such as glucose, insulin, cellular nutrients, and oxygen, while inhibiting the permeation of macromolecules such as antibodies and complement, or immune cells. For example, in the case of a semipermeable membrane such as Spectra / Por CE dialysis tubing (Repligen), which is a cellulose ester dialysis membrane, the cutoff value is referred to as "MWCO" and is sold in specifications such as 100 to 500 Da, 0.5 to 1 kDa, 3.5 to 5 kDa, 8 to 10 kDa, 20 kDa, 50 kDa, 100 kDa, 300 kDa, and 1000 kDa. The unit symbol for Dalton is Da, and 1,000 Da means 1 kDa.

[0074] In this specification, the thickness of the semipermeable membrane layer (semipermeable membrane) (SLT: 22 in Figure 2) can be considered to be the thickness of the layer obtained by subtracting the thickness of the nonwoven fabric layer (nonwoven fabric) (described below) (NLT: 21 in Figure 2) from the thickness of the biocompatible composite membrane (MT), and can be determined by SLT = MT - NLT. The thickness of the semipermeable membrane layer (semipermeable membrane) can be calculated, for example, by cutting the composite membrane to an arbitrary size from the biocompatible device, observing the cross section of the composite membrane under a microscope or the like, and subtracting the thickness of the nonwoven fabric from the thickness of the entire composite membrane.

[0075] The thickness (SLT) of the semipermeable membrane layer (semipermeable membrane) is not particularly limited, and examples thereof include 0 μm or more, about 0.1 μm or more, about 1.0 μm or more, about 5.0 μm or more, and about 10.0 μm or more. The upper limit of the thickness of the semipermeable membrane layer (semipermeable membrane) is not particularly limited, and examples thereof include about 500 μm or less, about 400 μm or less, about 300 μm or less, about 200 μm or less, and about 100 μm. However, SLT does not equal 0 μm.

[0076] The composite membrane of one embodiment of the present invention has a structure in which a constituent component of the semipermeable membrane (e.g., a cellulose derivative) partially penetrates into the nonwoven fabric, but as will be described later, the thickness of the portion where the constituent component of the semipermeable membrane partially penetrates into the nonwoven fabric (212 in Figure 2) is not included in the thickness of the semipermeable membrane layer (semipermeable membrane).

[0077] The semipermeable membrane layer can be formed, for example, by applying a solution containing a cellulose acetate derivative to one side of a nonwoven fabric and laminating it using the Robb-Srirajan method, as described in the Examples below.

[0078] In this specification, "penetration" and "penetrated" refer to partial penetration of the cellulose derivative (semipermeable membrane) from the surface of the nonwoven fabric to the interior. For details about penetration of the cellulose derivative, please refer to the description of "nonwoven fabric layer" below.

[0079] (Nonwoven Fabric Layer) The nonwoven fabric forming the nonwoven fabric layer is not particularly limited as long as it can serve as a support for the semipermeable membrane. Conventional semipermeable membranes have tended to be difficult to obtain sufficient strength against external forces such as tearing. Furthermore, when a cellulose derivative such as cellulose acetate is used as the semipermeable membrane, the high melting point of the cellulose derivative makes the thermocompression bonding of the membrane complicated, and the sealing and strength of the fused portion may be insufficient. In light of these circumstances, in the present invention, the strength of the composite membrane can be improved by forming a composite membrane by laminating the semipermeable membrane on the nonwoven fabric, using the nonwoven fabric as a support for the semipermeable membrane. Furthermore, the use of a thermoplastic nonwoven fabric improves the heat-sealing properties of the composite membrane, thereby suppressing or preventing leakage of cells or tissues from the device and maintaining the immunoisolation ability of the device.

[0080] Furthermore, the irregularities on the nonwoven fabric surface also function as anchors for the hydrogel encapsulated in the interior space of the biocompatible device (described below). This, for example, helps maintain the shape of the hydrogel in the interior space of the biocompatible device and prevents the gel from becoming distorted, folded, or collapsed in the interior space of the device. Without being bound by theory, this is thought to improve the utilization of cells or tissues embedded in the hydrogel and further enhance the therapeutic effect.

[0081] In one embodiment, the nonwoven fabric layer includes a portion where a semipermeable membrane component (e.g., a cellulose derivative) has partially penetrated the nonwoven fabric. That is, the nonwoven fabric layer of one embodiment of the present invention includes a portion where the semipermeable membrane component has not penetrated the nonwoven fabric and a portion where the semipermeable membrane component has partially penetrated the nonwoven fabric. The portion where the semipermeable membrane component has partially penetrated the nonwoven fabric is formed by immersing the nonwoven fabric in a solution of a cellulose derivative or the like that forms the semipermeable membrane layer, thereby allowing the semipermeable membrane component (e.g., a cellulose derivative) to penetrate into the mesh structure of the nonwoven fabric.

[0082] Thus, the composite membrane of one embodiment of the present invention is not a structure in which a semipermeable membrane and a nonwoven fabric are simply bonded together (e.g., a laminate structure), but rather has a structure in which the components of the semipermeable membrane partially penetrate the mesh structure of the nonwoven fabric and solidify, thereby integrating the semipermeable membrane and the nonwoven fabric. Therefore, the device of one embodiment of the present invention does not include a laminate membrane formed by lamination processing as the composite membrane. This makes it possible to prevent peeling, chipping, or damage to the semipermeable membrane, and to obtain a composite membrane that is stable over time.

[0083] In another embodiment of the present invention, the nonwoven fabric layer may have a semipermeable membrane component (e.g., a cellulose derivative) permeated throughout the entire nonwoven fabric. In this embodiment, the nonwoven fabric layer does not have a portion (211 in FIG. 2) that is not permeated by the semipermeable membrane component. In a preferred embodiment of the present invention, from the viewpoint of maintaining the thermal fusion properties of the nonwoven fabric layer, the nonwoven fabric layer includes a portion (211 in FIG. 2) that is not permeated by the semipermeable membrane component and a portion (212 in FIG. 2) where the semipermeable membrane component has partially permeated the nonwoven fabric.

[0084] As used herein, "fusion" refers to a state in which fibers are melted and bonded together. As used herein, "sealing" refers to, for example, a process in which two biocompatible composite membranes are stacked together with the semipermeable membrane layer disposed on the outer surface, and then a portion of the membranes several mm to several cm from the edge is heated to fuse the thermoplastic nonwoven fabric layer together, thereby forming a seal.

[0085] The nonwoven fabric can be made from a thermoplastic resin. Specific examples of such resins include polyolefin resins such as polyethylene (PE) and polypropylene (PP), polyester resins such as polyethylene terephthalate (PET), fluorine-containing resins such as polytetrafluoroethylene (PTFE), polyvinyl resins such as polystyrene (PS), and polyamide resins such as nylon. These resins may be used alone or in combination. In a preferred embodiment of the present invention, the nonwoven fabric contains a resin having a melting point in the range of about 90 to about 180°C, from the viewpoint of enhancing thermal adhesion. Among these, the nonwoven fabric preferably contains at least a polyolefin resin or polyester resin having a melting point in the range of about 90 to about 180°C, from the viewpoint of enhancing the strength of the device and the thermal adhesion of the composite membrane. For example, the nonwoven fabric preferably contains at least polyethylene.

[0086] Furthermore, the nonwoven fabric of one embodiment of the present invention may be formed from fibers containing two components with different melting points. Specifically, the nonwoven fabric of one embodiment of the present invention may be formed from, for example, core-sheath composite fibers. In a preferred embodiment of the present invention, the nonwoven fabric may be formed from heat-fusible core-sheath fibers in order to enhance heat-fusible properties. More specifically, such nonwoven fabrics include nonwoven fabrics formed from core-sheath fibers with polypropylene or polyethylene terephthalate as the core material and polyethylene as the sheath material.

[0087] From the above viewpoint, it is also preferable that the nonwoven fabric does not use a biodegradable or bioabsorbable resin. Examples of biodegradable or bioabsorbable resins include polylactic acid, polyglycolic acid, and poly-p-dioxane. Furthermore, the nonwoven fabric of one embodiment of the present invention is substantially free of an ethylene-vinyl alcohol copolymer. In one embodiment of the present invention, the content of the ethylene-vinyl alcohol copolymer in the components of the nonwoven fabric is less than about 0.1% by mass, less than about 0.05% by mass in another embodiment, and less than about 0.01% by mass in yet another embodiment, based on the total amount of the components of the nonwoven fabric.

[0088] In one embodiment of the present invention, the thickness of the nonwoven fabric layer (nonwoven fabric) refers to the thickness of the nonwoven fabric itself, regardless of the degree of penetration of the components of the semipermeable membrane. The thickness of the nonwoven fabric layer (nonwoven fabric) is specifically about 10 to about 1000 μm, in another embodiment about 50 to about 750 μm, and in yet another embodiment about 100 to about 500 μm. In addition, in one embodiment of the present invention, the basis weight (grammage) of the nonwoven fabric is about 5 to about 100 g / m ２ and in another embodiment, from about 10 to about 75 g / m ２ and in yet another embodiment, from about 15 to about 50 g / m ２ is.

[0089] (Biocompatible Composite Membrane) In one aspect of the present invention, the thickness of the biocompatible composite membrane can be the sum of the thickness of the nonwoven fabric itself and the thickness of the semipermeable membrane layer, regardless of the degree to which the components of the semipermeable membrane have penetrated into the nonwoven fabric. Specifically, the thickness of the biocompatible composite membrane is about 10 to about 1500 μm in one aspect, about 25 to about 800 μm in another aspect, about 25 to about 400 μm in yet another aspect, and about 100 to about 300 μm in yet another aspect.

[0090] The biocompatible composite membrane of the present invention is prepared, for example, by laminating a cellulose derivative on one side of a nonwoven fabric using the Robb-Srirajan method, as described in the Examples below, and is sometimes called an asymmetric porous membrane.

[0091] 1-2. Shape, Structure, etc. of the Biocompatible Device The biocompatible device of the present invention has an internal space formed by a biocompatible composite membrane including a semipermeable membrane layer and a nonwoven fabric layer. In this device, the semipermeable membrane layer is disposed on the outer side and the nonwoven fabric layer is disposed on the internal space side, and the internal space contains cells or tissues that secrete a biologically active substance. Furthermore, the biocompatible device of the present invention has an internal space formed by a biocompatible composite membrane in which a cellulose derivative is laminated on one side of a nonwoven fabric. In this device, the side of the biocompatible composite membrane on which the cellulose derivative is laminated is disposed on the outer side, and the internal space contains cells or tissues that secrete a biologically active substance.

[0092] For example, FIG. 1 is a conceptual diagram of a biocompatible device according to one embodiment of the present invention. In FIG. 1, the biocompatible device 1 has an internal space 3 formed by a biocompatible composite membrane 2. The internal space 3 is hollow so that cells or tissues 4 can be enclosed therein. The shape, volume, etc. of the internal space 3 are not particularly limited and can be appropriately designed depending on the intended use of the device 1. In one embodiment, the internal space 3 may be formed by partial fusion of a nonwoven fabric layer (nonwoven fabric) 21. In another embodiment, the cells or tissues 4 may be enclosed in the internal space 3 using a hydrogel 5 as a carrier. In another embodiment, the cells or tissues 4 do not need to use a hydrogel 5 as a carrier. In this case, the cells or tissues 4 can be enclosed in the internal space 3 in a state where they are mixed in a culture medium or the like.

[0093] Fig. 2 is a conceptual diagram showing a cross section of a biocompatible composite membrane 2 according to one embodiment of the present invention. In Fig. 2, the semipermeable membrane layer (semipermeable membrane) 22 constituting the biocompatible composite membrane 2 is disposed on the outer side of the biocompatible device 1, and the nonwoven fabric layer (nonwoven fabric) 21 is disposed on the inner space 3 side of the biocompatible device 1. As described above, the nonwoven fabric layer (nonwoven fabric) 21 may have a portion 211 where the semipermeable membrane constituent components have not penetrated, and a portion 212 where the semipermeable membrane constituent components have partially penetrated the nonwoven fabric.

[0094] The shape of the biocompatible device is not limited to the shape shown in Figure 1, as long as an internal space is formed by the biocompatible composite membrane. The method for forming the internal space by the biocompatible composite membrane is not particularly limited, but examples include a method in which two biocompatible composite membranes (e.g., square, rectangular, etc.) are prepared, placed face-to-face with the semipermeable membrane layer on the outside and the nonwoven fabric layer on the internal space side, and then fused and sealed at the edges (the fused portion is indicated by 6 in Figure 3). Another embodiment includes a method in which a single biocompatible composite membrane (e.g., square, rectangular, etc.) is prepared, folded, and placed face-to-face with the semipermeable membrane layer on the outside and the nonwoven fabric layer on the internal space side, and then fused and sealed at the edges. Although not shown, one embodiment of the device may have one or more internal spaces. Methods for forming multiple internal spaces include, for example, dividing one internal space into multiple spaces, or connecting multiple devices each having one internal space to form a single device having multiple internal spaces.

[0095] The shape of the device can be designed to a shape and size appropriate for the site of implantation. Specific examples of device shapes include flat plates. "Flat" refers to a flat plate having a substantially uniform thickness and a large area. Examples of such plate shapes include polygonal shapes such as triangles, rectangles, and pentagons, and circles. The thickness of a flat device is, in one embodiment, about 0.1 to about 100 mm, in another embodiment, about 0.2 to about 50 mm, in yet another embodiment, about 0.5 to about 20 mm, and in yet another embodiment, about 1 to about 20 mm. Furthermore, in one embodiment, the flat device has the aforementioned thickness and is substantially uniform throughout the entire plate. The thickness variation of a plate-shaped device is preferably within ±10%, more preferably within ±5%.

[0096] The thickness of a flat device is the thickness of the thickest part of the device. For example, the device may have a shape that at first glance resembles a rugby ball, with both ends being slightly thinner and the center being thicker than the ends. In such a shape, the thickness of the device means the thickness near the center, which is the thickest part. In some embodiments, the shape of a flat device is, for example, expressed as length x width x thickness, with a length of about 1 to about 200 mm, a width of about 1 to about 200 mm, and a thickness of about 0.2 to about 50 mm. In other embodiments, the flat device has an area, for example, expressed as length x width, of about 1 to about 40,000 mm. ２ , and a thickness of about 0.2 to about 50 mm, about 0.5 to about 50 mm, or about 1 to about 50 mm. In these embodiments, the device can be circular, square, hexagonal, octagonal, etc.

[0097] When the device is tubular, the shape of the device is not particularly limited as long as it is tubular or fibrous and elongated. A tubular shape is not limited to a circular cross-section perpendicular to the central axis, but may also be an asymmetrical structure or a deformed shape, for example, a polygonal cross-section such as a triangle, a square, or a pentagon, or an elliptical cross-section. In one embodiment, the cross-sectional shape is circular. Also included are tubes formed by connecting both ends to form a ring, or multiple such rings connected to form a chain, as well as tubes bundled together or connected in the form of a sheet.

[0098] When the device has a tubular shape, the diameter of its cross section (hereinafter, in the case of a non-circular device, this refers to the major axis or maximum diameter) is, for example, about 0.1 to about 100 mm in one embodiment, about 0.2 to about 50 mm in another embodiment, about 0.5 to about 20 mm in yet another embodiment, and about 1 to about 10 mm in yet another embodiment. Furthermore, when the device has a tubular shape, the length is not limited, but is, for example, about 10 mm to about 5 m in one embodiment, about 10 mm to less than 30 cm in another embodiment, and about 30 cm to about 5 m in yet another embodiment.

[0099] 1-3. Cells or Tissues Encapsulated in Biocompatible Devices Cells or tissues that can be encapsulated in biocompatible devices include, but are not limited to, cells or tissues that secrete bioactive substances (substances with pharmacological or biological activity). Bioactive substances are not particularly limited, but are preferably those used as pharmaceuticals, including, for example, small molecule drugs, medium molecule drugs such as peptides and oligonucleotides, biopharmaceuticals such as proteins, and physiologically active substances. As used herein, "biologically active substances" includes not only physiologically active substances inherent in living organisms (biologically active natural products), but also modified or altered bioactive substances, substances that activate or inhibit bioactivity, and the like. This term also includes naturally occurring substances as well as substances produced by genetic engineering or chemically synthesized, including prodrugs of these substances. Specific examples include vitamins, coenzymes, hormones, antibiotics, neurotransmitters, cytokines, enzymes, growth factors, antibodies, and other biological factors. More specific examples of biological factors include insulin, dopamine, factor VIII, factor IX, and the like. The cells secreting the bioactive substance include not only natural cells but also artificially modified cells, as well as cell masses consisting of multiple cells. The cells secreting the bioactive substance may also be transplantable cells. Examples of transplantable cells include insulin-secreting cells, pancreatic islets, and pancreatic islet cells. Examples of cells capable of secreting other bioactive substances include dopamine-secreting cells, pituitary cells, growth hormone-secreting cells, parathyroid cells, nerve growth factor-secreting cells, blood coagulation factor-secreting cells, hepatocytes, parathyroid cells, erythropoietin-secreting cells, and norepinephrine-secreting cells.

[0100] "Insulin-secreting cells" refer to cells that have the function of secreting insulin, such as beta cells that secrete insulin among cells that constitute pancreatic islets. Furthermore, "insulin-secreting cells" may also refer to cells that have acquired insulin secretion function through differentiation, maturation, modification, or the like, and may include, for example, cells with insulin secretion function obtained by differentiating stem cells such as iPS cells, ES cells, or somatic stem cells (e.g., mesenchymal stem cells), cells with insulin secretion function obtained by maturing immature cells or progenitor cells, and cells to which insulin secretion function has been imparted by genetic recombination. Here, differentiating or maturing the cells includes culturing the cells, and thus, cells obtained by differentiation or maturation may include cells obtained by culture.

[0101] A "pancreatic islet," also known as an islet of Langerhans, is a cell cluster consisting of an average of approximately 2,000 pancreatic islet cells. A pancreatic islet is composed of five types of cells: α cells that secrete glucagon, β cells that secrete insulin, δ cells that secrete somatostatin, ε cells that secrete ghrelin, and PP (pancreatic polypeptide) cells that secrete pancreatic polypeptide.

[0102] As used herein, the term "pancreatic islet cells" refers to cells that contain at least one of the five types of cells that constitute the pancreatic islets, and preferably includes at least β cells. In some embodiments, the pancreatic islet cells may be a mixture containing all of α cells, β cells, δ cells, ε cells, and PP cells, or may be cells contained in pancreatic islets.

[0103] Furthermore, "pancreatic islet cells" may be cells that have become pancreatic islet cells through differentiation, maturation, modification, etc. In this case, "pancreatic islet cells" may include, for example, pancreatic islet cells obtained by differentiating stem cells such as iPS cells, ES cells, and somatic stem cells (e.g., mesenchymal stem cells), as well as pancreatic islet cells obtained by maturing immature cells or progenitor cells.

[0104] When used for transplantation, "insulin-secreting cells" or "pancreatic islets (including pancreatic islet cells)" preferably have sufficient viability and function to recover a pathological condition of a patient when transplanted into the patient. The function of insulin-secreting cells, pancreatic islets, or pancreatic islet cells is, for example, to secrete insulin, and it is preferred that glucose responsiveness is maintained even after transplantation.

[0105] The donor of "insulin-secreting cells," "pancreatic islets," or "pancreatic islet cells" is an animal in one embodiment, a vertebrate in another embodiment, or a mammal in yet another embodiment, specifically a human, pig, monkey, rat, or mouse, and particularly a human or pig. In some embodiments, the donor of "insulin-secreting cells," "pancreatic islets," or "pancreatic islet cells" is a pig in order to alleviate the shortage of donors. "Insulin-secreting cells," "pancreatic islets," or "pancreatic islet cells" may be either pancreatic islets or pancreatic islet cells obtained from a donor animal, or insulin-secreting cells or pancreatic islet cells obtained from donor-derived cells, and may be, for example, insulin-secreting cells or pancreatic islet cells differentiated from human-derived ES cells or iPS cells.

[0106] When the "insulin-secreting cells," "pancreatic islets," or "pancreatic islet cells" are derived from pigs, examples include adult porcine pancreatic islets, or fetal, neonatal, or perinatal porcine pancreatic islets, or insulin-secreting cells or pancreatic islet cells obtained from such pancreatic islets. The pancreatic islets may be appropriately cultured before use, or islets obtained by maturing fetal, neonatal, or perinatal porcine pancreatic islets may also be used.

[0107] Examples of blood coagulation factor-secreting cells include factor VIII-secreting cells and factor IX-secreting cells.

[0108] Other cells for encapsulation include animal cells in one embodiment, vertebrate-derived cells in another embodiment, and mammalian-derived cells in yet another embodiment, particularly human-derived or porcine-derived cells. The vertebrate-derived cells (particularly human-derived cells) may be stem cells (e.g., pluripotent stem cells (mutilative cells) or somatic stem cells), progenitor cells, or mature cells. Examples of pluripotent stem cells include embryonic stem (ES) cells, germline stem (GS) cells, and induced pluripotent stem (iPS) cells. Examples of somatic stem cells include mesenchymal stem cells (MSCs), hematopoietic stem cells, amniotic cells, umbilical cord blood cells, bone marrow-derived cells, cardiac muscle stem cells, adipose-derived stem cells, and neural stem cells. These stem cells may also be differentiated and used.

[0109] Examples of progenitor cells and mature cells that can be used include cells derived from skin, dermis, epidermis, muscle, cardiac muscle, nerve, bone, cartilage, endothelium, brain, epithelium, heart, kidney, liver, spleen, oral cavity, cornea, bone marrow, umbilical cord blood, amniotic membrane, and hair. Examples of human-derived cells that can be used include ES cells, iPS cells, MSCs, chondrocytes, osteoblasts, osteoblast precursor cells, mesenchymal cells, myoblasts, cardiac muscle cells, cardiac muscle blasts, neurons, hepatocytes, fibroblasts, corneal endothelial cells, vascular endothelial cells, corneal epithelial cells, amniotic cells, umbilical cord blood cells, bone marrow-derived cells, and hematopoietic stem cells. Furthermore, the origin of the cells may be either autologous or allogeneic.

[0110] In some embodiments, the biocompatible device may contain a substance for maintaining the viability and function of cells or tissues, a substance (such as a hydrogel) that allows cells or tissues secreting bioactive substances to be uniformly dispersed, etc. The device of the present invention encompasses both embodiments that contain these and embodiments that do not.

[0111] In some embodiments, cells or tissues are encapsulated in an internal space using a hydrogel as a carrier. As used herein, "hydrogel" refers to a water-insoluble polymer with a three-dimensional network structure and its water-swollen form. In this specification, hydrogel may be simply referred to as "gel."

[0112] In one embodiment of the present invention, materials used to form hydrogels include alginic acid (including alginate esters and alginates, as described below), collagen, hyaluronan, gelatin, fibronectin, elastin, tenascin, laminin, vitronectin, polypeptides, heparan sulfate, chondroitin, chondroitin sulfate, keratan, keratan sulfate, dermatan sulfate, carrageenan, heparin, chitin, chitosan, agarose, agar, cellulose, methylcellulose, carboxymethylcellulose, glycogen and derivatives thereof, as well as fibrin, fibrinogen, thrombin, polyglutamic acid, polylactic acid, polyglycolic acid, lactic acid-glycolic acid copolymer, gellan gum, xanthan gum, galactomannan, guar gum, locust bean gum, and tara gum. These materials may be used alone or in combination of two or more. Furthermore, these materials can be hydrogelated by heat, temperature, light, or chemical (e.g., ionic crosslinking, chemical crosslinking, etc.) action. In another aspect of the present invention, the material used to form the hydrogel may be alginic acid or a derivative thereof. In this case, it is preferable to use alginic acid or a derivative thereof modified so that it can gel by chemical crosslinking. Specific examples of chemically modified alginic acid derivatives include those described in Aspects [3-5] to [3-9] and [1A-9-2] to [1A-11] above. Furthermore, examples of alginic acid with chemical crosslinking (chemically crosslinked alginic acid) include those described in Aspects [3-10] to [3-13] and [3A] to [3A-5] above. By including alginic acid, or in some aspects, chemically crosslinked alginic acid, as the hydrogel, the network structure of alginic acid or chemically crosslinked alginic acid can be more firmly bound to the irregularities on the surface of the nonwoven fabric, thereby further improving the anchoring effect of the nonwoven fabric.

[0113] The hydrogel of one embodiment of the present invention is substantially free of a vinyl alcohol-based polymer. In one embodiment of the present invention, the content of the vinyl alcohol-based polymer in the hydrogel is less than about 0.1% by mass, less than about 0.05% by mass in another embodiment, and less than about 0.01% by mass in yet another embodiment, based on the total amount of the components of the hydrogel.

[0114] Hereinafter, alginic acid and alginic acid derivatives, which are raw materials for the chemically crosslinked alginic acid that constitutes the hydrogel in one embodiment of the device, will be described.

[0115] 2. Alginic Acid In this specification, the term "alginic acid" refers to at least one type of alginic acid (sometimes referred to as "alginic acids") selected from the group consisting of alginic acid, alginate esters, and salts thereof (e.g., sodium alginate). The alginic acid used may be naturally occurring or synthetic, but is preferably naturally occurring.

[0116] In this specification, alginic acid may be expressed as (ALG)-COOH, where alginic acid is (ALG) and any one of the carboxyl groups of alginic acid is -COOH.

[0117] In some embodiments, the alginic acid is sodium alginate. Commercially available sodium alginate can be used as the sodium alginate. In the examples described below, sodium alginate A-1, A-2, A-3, B-1, B-2, and B-3 (distributed by Mochida Pharmaceutical Co., Ltd.) shown in the table below is used as the sodium alginate. The viscosity, weight-average molecular weight, and M / G ratio of a 1 w / w% aqueous solution of each sodium alginate are shown in the table below.

[0118]

[0119] In this specification, the terms "alginate ester" and "alginate salt" are not particularly limited, but are required to have no functional groups that inhibit the crosslinking reaction. Preferred examples of alginate esters include propylene glycol alginate.

[0120] In this specification, examples of alginate salts include monovalent alginic acid salts and divalent alginic acid salts. Preferred examples of monovalent alginic acid salts include sodium alginate, potassium alginate, and ammonium alginate, more preferably sodium alginate or potassium alginate, and particularly preferably sodium alginate. Preferred examples of divalent alginic acid salts include calcium alginate, magnesium alginate, barium alginate, and strontium alginate.

[0121] For other explanations regarding alginic acid, see PCT / JP2019 / 023478 (filed June 13, 2019), PCT / JP2020 / 047100 (filed December 17, 2020), and PCT / JP2019 / 007655 (filed February 27, 2019).

[0122] 3. Alginic Acid Derivative In some embodiments, the alginic acid derivative in the present specification is an alginic acid derivative in which a reactive group in the Huisgen reaction or a reactive group complementary to the reactive group is introduced to any one or more carboxyl groups of alginic acid via an amide bond and a divalent linker. More specifically, the alginic acid derivative is represented by the following formula (IA): [In formula (IA), (ALG), -NHCO-, -L １Ａ - and Aky are defined as in [1A-9-2] above], and an alginic acid derivative represented by the following formula (II-A): [In formula (II-A), (ALG), —NHCO—, and —L ２Ａ - is the same as defined in [1A-9-2] above], and is an alginic acid derivative represented by the following formula (HI): [In formula (HI), (ALG) represents alginic acid; -NHCO- represents an amide bond via any carboxyl group of alginic acid; -L １ - is a divalent linker that bonds to a cyclic alkyne group (Akn)], and an alginic acid derivative represented by the following formula (H-II): [In formula (H-II), (ALG) represents alginic acid; -NHCO- represents an amide bond via any carboxyl group of alginic acid; -L ２ - is a divalent linker that bonds to the azide group. １ -or-L ２ Any straight-chain group can be used for the reactive group (-(CH ２ ) ｎ -, n = 1 to 30) (-CH ２ - may be replaced by a plurality of (for example, 1 to 10, or 1 to 5) groups such as -C(═O)-, -CONH-, -O-, -NH-, -S-, a benzene ring, or a heterocycle (a 5- or 6-membered aromatic heterocycle or a 5- or 6-membered non-aromatic heterocycle such as a pyridine ring, a piperidine ring, or a piperazine ring), and the -CH ２ The hydrogen atom of - is an oxo group (=O), C １－６ Examples of the alkyl group include, but are not limited to, alkyl groups (e.g., methyl groups, ethyl groups, n-propyl groups, isopropyl groups, etc.), halogen atoms (e.g., fluorine atoms, chlorine atoms, bromine atoms, iodine atoms, etc.), hydroxyl groups (—OH), etc., which may be substituted with a plurality of groups (e.g., 1 to 10 groups, or 1 to 5 groups).

[0123] In the -NH-CO- group of the chemically modified alginic acid derivative represented by the formula (IA), formula (II-A), formula (HI) or formula (H-II), the hydrogen atom of the imino group (-NH-) can be substituted with a methyl group to form an -N(Me)-CO- group. In the chemically modified alginic acid derivative represented by the formula (HI) or formula (H-II), the linker (-L １ -, -L ２ The bonding mode between the linker (-L) and alginic acid, or the chemically modified alginic acid derivative represented by the formula (IA) or (II-A), １Ａ -, -L ２ＡThe bond between the alginic acid and the hydroxyl group is an —NH—CO— bond or an —N(Me)—CO— bond, preferably an —NH—CO— bond. The —CO— in the —NH—CO— bond or the —N(Me)—CO— bond is derived from the carboxyl group of alginic acid.

[0124] In this specification, unless otherwise specified, the term "cyclic alkyne group" means a "5- to 9-membered cycloalkyne group", and the -CH ２ The cyclic alkyne group also includes a cycloalkyne group in which - is substituted with 1 to 4 groups selected from the group consisting of -NH-, -S-, -O-, or =C(=O). ２ - hydrogen atom is a halogen atom, a hydroxyl group, an amino group, a keto group, C １－３ Alkyl group, —O—C １－３ Alkyl group, —NH(C １－３ alkyl group), -N(C １－３ alkyl group) ２ , -COO-M (M = Na, K, 1 / 2 Ca, hydrogen atom, C １－３ The cyclic alkyne group may be substituted with 1 to 5 groups selected from groups such as C 3-8 The ring may be condensed with 1 to 3 rings selected from a cycloalkyl ring, a benzene ring, and a 5- or 6-membered aromatic heterocycle.

[0125] In this specification, unless otherwise specified, the term "5- to 9-membered cycloalkyne group" refers to a monocyclic saturated cycloalkyl group having 5 to 9 carbon atoms, -CH ２ -CH ２ - means a group in which - is replaced by -C≡C-, and examples thereof include a cyclopentyne group, a cyclohexyne group, a cycloheptyne group, a cyclooctyne group, and a cyclononyne group. ２ The - may be substituted with 1 to 4 groups selected from the group consisting of -NH-, -S-, -O-, or =C(=O).

[0126] In this specification, unless otherwise specified, the cyclic alkyne group is preferably a 7- to 9-membered cyclic alkyne group (the —CH ２- also includes a cycloalkyne group substituted with 1 to 4 groups selected from the group consisting of -NH-, -S-, -O-, or =C(=O); -CH on the ring of a 7- to 9-membered cyclic alkyne group ２ - hydrogen atom is a halogen atom, a hydroxyl group, an amino group, a keto group, C １－３ Alkyl group, —O—C １－３ Alkyl group, —NH(C １－３ alkyl group), -N(C １－３ alkyl group) ２ , -COO-M (M = Na, K, 1 / 2 Ca, hydrogen atom, C １－３ The 7- to 9-membered cyclic alkyne group may be substituted with 1 to 5 groups selected from groups such as C 3-8 It may be fused with 1 to 3 rings selected from a cycloalkyl ring, a benzene ring, and a 5- or 6-membered aromatic heterocycle; more preferably, it is an 8-membered cyclic alkyne group (cyclooctyne group) (the —CH ２ - also includes a cycloalkyne group substituted with 1 to 4 groups selected from the group consisting of -NH-, -S-, -O-, or =C(=O); -CH on the ring of the cyclooctyne group ２ - hydrogen atom is a halogen atom, a hydroxyl group, an amino group, a keto group, C １－３ Alkyl group, —O—C １－３ Alkyl group, —NH(C １－３ alkyl group), -N(C １－３ alkyl group) ２ , -COO-M (M = Na, K, 1 / 2 Ca, hydrogen atom, C １－３ The cyclooctyne group may be substituted with 1 to 5 groups selected from groups such as C 3-8 and a cycloalkyl ring, a benzene ring, or a 5- or 6-membered aromatic heterocycle, each of which may be condensed with 1 to 3 rings; more preferably, a cycloalkyl ring, a benzene ring, or a 5- or 6-membered aromatic heterocycle; [wherein the right side of the dashed lines at both ends is not included]; particularly preferably, a group selected from the following partial structural formula: [wherein the right side of the dashed lines at both ends is not included]; most preferably, a group selected from the following partial structural formula: [wherein the right side of the dashed lines at both ends is not included].

[0127] In one embodiment of the alginic acid derivative, in the formula (IA) or (HI), -L １Ａ -or-L １ - is - (CH ２ ) ｎ１ - (where n1=1 to 50, and -CH ２ - may be replaced by 1 to 10 groups selected from -CO-, -CONH-, -NHCO-, -O-CONH-, -NHCO-O-, -O- and -NH-, or a benzene ring, ２ The hydrogen atom of - is C １－3 C substituted with alkyl or phenyl groups １－3 Aky or Akn is an 8-membered cyclic alkyne group (wherein the cyclic alkyne group may be further fused with 1 or 2 benzene rings, cyclopropane rings or 1,2,3-triazole rings, and in the fused ring, -L １ -, and -CH ２ - may be replaced by 1 or 2 groups selected from -C(=O)-, -CONH-, and -NH-, and the -CH ２ The hydrogen atom of - is C １－３ Alkyl group, fluorine atom, hydroxyl group, C １－３ and the compound represented by formula (II-A) or (H-II) is a compound represented by formula (II-B) or (H-II), wherein -L ２Ａ -or-L ２ -ga- (CH ２ ) ｎ２ - (where n2=1 to 50, and -CH ２ - may be replaced by 1 to 10 groups selected from -CONH-, -NHCO-, -O- and -NH-, or a benzene ring or a pyridine ring, and the -CH ２ The hydrogen atom of - is C １－3 The compound is a compound in which the formula (I) is a substituted or unsubstituted alkyl group.

[0128] In the above compounds, n1 is preferably 1 to 20, and more preferably 1 to 15 (for example, 2 to 15). １Ａ-or-L １ A preferred embodiment of - is -(CH ２ ) ｎ１ -Medium, -CH ２ - may be replaced by 1 to 5, more preferably 1 to 3, groups selected from -CO-, -CONH-, -NHCO-, -O-CONH-, -NHCO-O-, -O-, and -NH-. Aky or Akn is preferably a group in which one or two benzene rings may be condensed, and -CH in the 8-membered cyclic alkyne group ２ n2 is an 8-membered cyclic alkyne group in which - may be replaced by -NH-. In a preferred embodiment, n2 is 1 to 20, and in a more preferred embodiment, it is 1 to 15 (for example, 2 to 15). ２Ａ -or-L ２ A preferred embodiment of - is -(CH ２ ) ｎ２ -Medium, -CH ２ - is a group which may be substituted by 1 to 5, more preferably 1 to 3, groups selected from -CONH-, -NHCO-, -O- and -NH-.

[0129] A preferred embodiment of the alginic acid derivative is the alginic acid derivative represented by the formula (IA) above, wherein -L １Ａ - or in the formula (HI), -L １ - is - (CH ２ ) ｎ１ - (where n1=1 to 15 (e.g., 2 to 15), and -CH ２ - may be replaced by 1 to 5 groups selected from -CO-, -CONH-, -NHCO-, -O-CONH-, -NHCO-O-, -O- and -NH-, or a benzene ring, ２ The hydrogen atom of - is C １－3 C substituted with alkyl or phenyl groups １－3 Aky or Akn is an 8-membered cyclic alkyne group (wherein one or two benzene rings may be further condensed to the cyclic alkyne group, and the —CH ２ In the formula (II-A), -L may be an 8-membered ring group in which - is replaced by -NH-.２Ａ - or in the formula (H-II), -L ２ - is - (CH ２ ) ｎ２ - (where n2=1 to 15 (e.g., 2 to 15), and -CH ２ - may be replaced by 1 to 5 groups selected from -CONH-, -NHCO-, -O- and -NH-, or a benzene ring.

[0130] Unless otherwise specified, in this specification, examples of the "halogen atom" include a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom.

[0131] In this specification, unless otherwise specified, 1-３ Examples of the "alkyl group" include methyl, ethyl, propyl, and isopropyl groups.

[0132] In this specification, unless otherwise specified, ２－４ The "alkanoyl group" refers to the group defined above as "C １－３ A carbonyl group is bonded to an alkyl group. １－３ alkylcarbonyl group, and examples thereof include acetyl, propionyl, butyryl, and the like.

[0133] In this specification, unless otherwise specified, 3-8 The "cycloalkyl ring" includes monocyclic or polycyclic saturated or unsaturated cycloalkyl rings having 3 to 8 carbon atoms, such as cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, and cyclooctane rings.

[0134] In this specification, unless otherwise specified, ５－９ The "cycloalkene ring" includes monocyclic cycloalkene rings having 5 to 9 carbon atoms, such as cyclopentene, cyclohexene, cycloheptene, cyclooctene, and cyclononene rings.

[0135] In this specification, unless otherwise specified, the term "5- or 6-membered aromatic heterocycle" means a 5- or 6-membered unsaturated ring containing 1 to 4 heteroatoms selected from the group consisting of nitrogen atoms, sulfur atoms, and oxygen atoms.

[0136] Unless otherwise specified, in this specification, examples of the "5- to 6-membered aromatic heterocycle" include pyrrole, furan, thiophene, imidazole, pyrazole, oxazole, isoxazole, thiazole, isothiazole, triazole, oxadiazole, furazan, thiadiazole, tetrazole, pyridine, pyridazine, pyrimidine, pyrazine, triazine, thiadiazine, and the like.

[0137] In this specification, unless otherwise specified, the term "5- to 6-membered non-aromatic heterocycle" refers to a 5- to 6-membered saturated heterocycle containing 1 to 4 heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom. In this specification, unless otherwise specified, examples of the "5- to 6-membered non-aromatic heterocycle" include rings such as pyrrolidine, tetrahydrofuran, thiolane, piperidine, dihydropyran, tetrahydropyran, tetrahydrothiopyran, piperazine, dioxane, morpholine, thiomorpholine, and quinuclidine.

[0138] In the present specification, the alginic acid derivatives represented by the formula (IA), formula (II-A), formula (HI) or formula (H-II) can be produced, for example, by the synthesis method of alginic acid derivatives described below.

[0139] In one embodiment of the alginic acid derivative, in the formula (HI), -L １ In one embodiment of the alginic acid derivative, in the formula (H-II), -L ２ - is a compound in which the definition is the same as that in the above embodiment [3-8].

[0140] In one embodiment of the alginic acid derivative, in the formula (IA), -L １Ａ In one embodiment of the alginic acid derivative, in the formula (II-A), -L ２Ａ - is a compound in which the definition of - is the same as that of the above-mentioned embodiment [1A-9-2].

[0141] In the present specification, specific examples of the alginic acid derivatives represented by the formula (IA) or (HI) include those represented by the following formula: Examples of alginic acid derivatives include, but are not limited to, those represented by the following formula:

[0142] In the present specification, specific examples of the alginic acid derivatives represented by the formula (II-A) or (H-II) include those represented by the following formula: Examples of alginic acid derivatives include, but are not limited to, those represented by the following formula:

[0143] Unless otherwise specified, in this specification, when a variable substituent is substituted on a cyclic group, the variable substituent is not bonded to a specific carbon atom of the cyclic group. For example, the variable substituent Rs in the following formula A means that it can be substituted on any of carbon atoms i, ii, iii, iv, or v in the formula A.

[0144]

[0145] The weight-average molecular weight of the alginic acid derivatives represented by formula (IA) or formula (II-A) herein, and the alginic acid derivatives represented by formula (HI) or formula (H-II) herein is about 100,000 Da to about 3,000,000 Da, preferably about 300,000 Da to about 2,500,000 Da, and more preferably about 500,000 Da to about 2,000,000 Da. The molecular weight of the alginic acid derivative can be determined by the methods described in PCT / JP2019 / 023478 (filed June 13, 2019) and PCT / JP2020 / 047100 (filed December 17, 2020).

[0146] In the present specification, Aky-L of formula (IA) or formula (HI) １Ａ -NH- group or Akn-L １ The —NH— group does not necessarily have to be bonded to all the carboxyl groups of the alginic acid structural unit, and the N ３ -L ２Ａ -NH- group or N ３ -L ２ It is not necessary for the --NH-- group to be bonded to all of the carboxyl groups of the alginic acid building block.

[0147] In the present specification, Aky-L of formula (IA) or formula (HI) １Ａ -NH- group or Akn-L １ When the —NH— group is referred to as a reactive group, it is the N ３ -L ２Ａ -NH- group or N ３ -L ２ The —NH— group serves as a complementary reactive group. ３ -L ２Ａ -NH- group or N ３ -L ２ When the —NH— group is referred to as a reactive group, it is Aky-L of formula (IA) or formula (HI). １Ａ -NH- group or Akn-L １ The —NH— group serves as a complementary reactive group.

[0148] In this specification, the introduction rate of the reactive group or the complementary reactive group is about 0.1% to about 30%, or about 1% to about 30%, preferably about 2% to about 20%, and more preferably about 3% to about 10%, respectively.

[0149] The introduction rate of the reactive group or complementary reactive group is the number of uronic acid monosaccharide units into which each reactive group has been introduced, expressed as a percentage, among the uronic acid monosaccharide units, which are the repeating units of alginic acids. Unless otherwise specified, the percentage used for the introduction rate of the reactive group or complementary reactive group in an alginic acid derivative (formula (IA), formula (II-A), formula (HI), or formula (H-II)) refers to mol%. The introduction rate of each reactive group or complementary reactive group can be determined by the methods described in PCT / JP2019 / 023478 (filed June 13, 2019) and PCT / JP2020 / 047100 (filed December 17, 2020).

[0150] In this specification, the cyclic alkyne group (Akn) in formula (H-I) and the azide group in formula (H-II) form a triazole ring by a Huisgen reaction, thereby forming a crosslink. Furthermore, the cyclic alkyne group (Aky) in formula (IA) and the azide group in formula (II-A) form a triazole ring by a Huisgen reaction, thereby forming a crosslink. For details of the Huisgen reaction, see PCT / JP2019 / 023478 (filed June 13, 2019) and PCT / JP2020 / 047100 (filed December 17, 2020).

[0151] 4. Method for synthesizing alginic acid derivatives The alginic acid derivatives represented by formula (IA), formula (II-A), formula (HI), and formula (H-II) can be prepared, for example, by the following production method (Aky, -L in each formula). １Ａ -, -L ２Ａ For the definitions of -, (ALG), etc., see above; Aky is Akn, -L １Ａ -Ha-L １ - to, -L ２Ａ -Ha-L ２ - may be replaced by ). For more detailed reaction conditions, please refer to the general production methods and examples in PCT / JP2019 / 023478 (filed June 13, 2019) and PCT / JP2020 / 047100 (filed December 17, 2020). By referring to these, the alginic acid derivatives described in this specification can be produced.

[0152] 5. Cross-linked Alginic Acid Cross-linked alginic acid can be (i) via a divalent metal ion bond, (ii) via a chemical bond, or (iii) via both a divalent metal ion bond and a chemical bond. All cross-linked alginic acids have the property of forming a gel-like to semi-solid, and in some cases, a sponge-like form. Examples of divalent metal ions include, but are not limited to, calcium ions, magnesium ions, barium ions, strontium ions, and zinc ions. In one embodiment, the cross-linked alginic acid is calcium ions or barium ions.

[0153] Crosslinked alginate via divalent metal ion bonds undergoes an ultrafast reaction that is reversible, whereas crosslinked alginate via chemical bonds undergoes a slow reaction under relatively mild conditions and is irreversible. The physical properties of crosslinked alginate can be adjusted, for example, by changing the concentration of the aqueous solution containing the divalent metal ion (e.g., calcium chloride aqueous solution, barium chloride aqueous solution, etc.) or the introduction rate of reactive groups introduced into alginate. The divalent metal ion concentration (e.g., calcium ion or barium ion concentration) of the aqueous solution containing the divalent metal ion is not particularly limited, but is, for example, in one embodiment, in the range of about 1 mmol / L to about 1 mol / L, in another embodiment, in the range of about 2 mmol / L to about 500 mmol / L, and in yet another embodiment, in the range of about 5 mmol / L to about 100 mmol / L. In this specification, an aqueous solution containing a divalent metal ion may be simply referred to as a divalent metal ion solution, and unless otherwise specified, this divalent metal ion solution refers to an aqueous solution.

[0154] By utilizing the crosslinking reaction, it is possible to prepare various alginate structures (e.g., hydrogels). For example, specific structures can be instantly created from an alginate solution by ionic crosslinking, and crosslinking reactions using chemical bonds can be used to strengthen the structure (e.g., to achieve long-term stability, etc.). Furthermore, for example, in alginate structures crosslinked via both divalent metal ion bonds and chemical bonds, the divalent metal ions incorporated by ionic crosslinking can be reversibly released, creating a structure in which only crosslinks via chemical bonds remain. A preferred embodiment of crosslinked alginate includes chemically crosslinked alginate obtained by crosslinking reactions using chemical bonds. Another preferred embodiment of crosslinked alginate includes the above-mentioned chemically crosslinked alginate and ionically crosslinked alginate via divalent metal ion bonds. Yet another preferred embodiment of crosslinked alginate includes the above-mentioned chemically crosslinked alginate and, optionally, the above-mentioned ionically crosslinked alginate.

[0155] A crosslinked alginic acid in one embodiment can be obtained by mixing alginic acid derivatives of the formula (H-I) and the formula (H-II) and then carrying out a Huisgen reaction. Alternatively, a crosslinked alginic acid in one embodiment can be obtained by mixing alginic acid derivatives of the formula (IA) and the formula (II-A) and then carrying out a Huisgen reaction. The crosslinked alginic acid in one embodiment forms a three-dimensional network structure via chemical crosslinking (crosslinking by a triazole ring formed from an alkyne group and an azide group).

[0156] In some embodiments, the crosslinked alginic acid has a bond between any carboxyl group of a first alginic acid and any carboxyl group of a second alginic acid represented by the following formula (H-III-L): [In formula (H-III-L), -CONH- and -NHCO- at both ends represent an amide bond via any carboxyl group of alginic acid; -L １ -, -L ２ -, and X is the same as defined in any one of the above embodiments [3-10] to [3-13], and is a cross-linked alginic acid bonded via an amide bond.

[0157] In some embodiments, when preparing crosslinked alginic acid, the mixing ratio of the alginic acid derivative of formula (H-I) to the alginic acid derivative of formula (H-II) is, for example, 1 to 1.5:1, preferably 1.2 to 1.5:1, or 1 to 1.2:1, more preferably 1:1, in terms of the weight ratio of the derivative of formula (H-I) to the derivative of formula (H-II).

[0158] In some embodiments, the mixing ratio of the alginic acid derivative of formula (H-II) to the alginic acid derivative of formula (H-I) when preparing the crosslinked alginic acid is, for example, 1 to 4.0:1, preferably 1.5 to 4.0:1, or 1.2 to 1.5:1, or 1 to 1.2:1, more preferably 1:1, in terms of the weight ratio of the derivative of formula (H-II) to the derivative of formula (H-I).

[0159] In some embodiments, when preparing crosslinked alginic acid, the mixing ratio of the alginic acid derivative of formula (H-I) to the alginic acid derivative of formula (H-II) is, more preferably, the ratio of the introduction rate (mol%) of the reactive group of the alginic acid derivative of formula (H-I) to the alginic acid derivative of formula (H-II), for example, 1 to 1.5:1, preferably 1.2 to 1.5:1, or 1 to 1.2:1, more preferably 1:1.

[0160] In some embodiments, when preparing a crosslinked alginic acid, the mixing ratio of the alginic acid derivative of formula (H-II) to the alginic acid derivative of formula (H-I) is, more preferably, the ratio of the introduction rate (mol%) of the reactive group of the alginic acid derivative of formula (H-II) to the alginic acid derivative of formula (H-I), for example, 1 to 4.0:1, preferably 1.5 to 4.0:1, or 1.2 to 1.5:1, or 1 to 1.2:1, more preferably 1:1.

[0161] In addition, in the above mixing ratio, the alginic acid derivative of formula (HI) can be replaced with the alginic acid derivative of formula (H-II), and the alginic acid derivative of formula (H-II) can be replaced with the alginic acid derivative of formula (HI).

[0162] In crosslinked alginic acid, it is not necessary for all carboxyl groups in the constituent units of alginic acid to have crosslinks of the above formula (H-III-L). The introduction rate of crosslinks represented by the above formula (H-III-L) in crosslinked alginic acid (also referred to as crosslinking rate) is, for example, in the range of about 0.1 to about 80%, about 0.3 to about 60%, about 0.5 to about 30%, or about 1.0 to about 10%.

[0163] The concentration of the alginic acid derivative of formula (HI) or (H-II) in the Huisgen reaction to obtain crosslinked alginic acid is usually in the range of about 1 to about 500 mg / mL, preferably about 5 to about 100 mg / mL.

[0164] In some embodiments, the crosslinked alginic acid has a bond between any carboxyl group of the first alginic acid and any carboxyl group of the second alginic acid represented by the following formula (III-Lx): [In formula (III-Lx), —NHCO— on the right end represents an amide bond derived from the alginic acid derivative of formula (IA), and —CONH— on the left end represents an amide bond derived from the alginic acid derivative of formula (II-A); １Ａ - and -L ２Ａ - is the same as defined in the above [1A-9-2]; -X Ａ - is cross-linked alginic acid bonded via an amide bond through the same definition as in [3A] above.

[0165] In some embodiments, when preparing crosslinked alginic acid, the mixing ratio of the alginic acid derivative of formula (IA) to the alginic acid derivative of formula (II-A) is, for example, 1 to 1.5:1, preferably 1.2 to 1.5:1, or 1 to 1.2:1, more preferably 1:1, by weight of the derivative of formula (IA) to the derivative of formula (II-A).

[0166] In some embodiments, the mixing ratio of the alginic acid derivative of formula (II-A) to the alginic acid derivative of formula (IA) when preparing the crosslinked alginic acid is, for example, 1 to 4.0:1, preferably 1.5 to 4.0:1, or 1.2 to 1.5:1, or 1 to 1.2:1, more preferably 1:1, in terms of the weight ratio of the derivative of formula (II-A) to the derivative of formula (IA).

[0167] In some embodiments, the mixing ratio of the alginic acid derivative of formula (IA) to the alginic acid derivative of formula (II-A) when preparing the crosslinked alginic acid is, more preferably, the ratio of the introduction rate (mol%) of the reactive group of the alginic acid derivative of formula (IA) to the alginic acid derivative of formula (II-A) is, for example, 1 to 1.5:1, preferably 1.2 to 1.5:1, or 1 to 1.2:1, more preferably 1:1.

[0168] In some embodiments, when preparing crosslinked alginic acid, the mixing ratio of the alginic acid derivative of formula (II-A) to the alginic acid derivative of formula (IA) is, more preferably, the ratio of the introduction rate (mol%) of the reactive group of the alginic acid derivative of formula (II-A) to the alginic acid derivative of formula (IA) is, for example, 1 to 4.0:1, preferably 1.5 to 4.0:1, or 1.2 to 1.5:1, or 1 to 1.2:1, more preferably 1:1.

[0169] In addition, in the above mixing ratio, the alginic acid derivative of formula (IA) can be replaced with the alginic acid derivative of formula (II-A), and the alginic acid derivative of formula (II-A) can be replaced with the alginic acid derivative of formula (IA).

[0170] In crosslinked alginic acid, it is not necessary for all carboxyl groups in the constituent units of alginic acid to have crosslinks of the formula (III-Lx). The introduction rate of crosslinks represented by the formula (III-Lx) (also referred to as crosslinking rate) in crosslinked alginic acid is, for example, in the range of about 0.1 to about 80%, about 0.3 to about 60%, about 0.5 to about 30%, or about 1.0 to about 10%.

[0171] The concentration of the alginic acid derivative of formula (IA) or formula (II-A) in the Huisgen reaction to obtain crosslinked alginic acid is usually in the range of about 1 to about 500 mg / mL, preferably about 5 to about 100 mg / mL.

[0172] The reaction temperature for the Huisgen reaction is usually in the range of about 4 to about 60°C, preferably about 15 to about 40°C.

[0173] The stirring time for forming crosslinked alginic acid is, for example, from a few seconds to about 24 hours, from a few seconds to about 12 hours, from a few seconds to about 30 minutes, or from a few seconds to about 10 minutes.

[0174] The reaction solvent or reaction solution used in the Huisgen reaction is not particularly limited, and examples thereof include tap water, pure water (e.g., distilled water, ion-exchanged water, RO water, RO-EDI water, etc.), ultrapure water, cell culture medium, phosphate buffered saline (PBS), and saline, with ultrapure water being preferred.

[0175] The cross-linked alginate in some embodiments is a cross-linked alginate containing chemical cross-links via triazole rings formed by the Huisgen reaction and ionic cross-links partially formed by calcium ions or barium ions.

[0176] 6. Method for Producing a Biocompatible Device The method for producing a biocompatible device will be described in more detail. The method for producing a biocompatible device according to one aspect of the present invention may be, for example, the method described in "6-1" or "6-2" below.

[0177] 6-1. Manufacturing Method of Biocompatible Device (1) One embodiment of the manufacturing method includes the following steps (a) to (d): Step (a): A step of preparing a semipermeable membrane layer formed from a cellulose derivative using a nonwoven fabric as a support, thereby forming a biocompatible composite membrane having a structure in which the cellulose derivative partially penetrates the nonwoven fabric. Step (b): A step of mixing cells or tissues (e.g., cells or tissues secreting a biologically active substance) with a hydrogelable solution (e.g., an alginate solution, or a gellable solution described in [3-1], [3-2], or [1A-9] to [1A-11] above). Step (c): A step of gelling the solution obtained in step (b). Step (d): A step of covering the hydrogel obtained in step (c), in which the cells or tissues are embedded, with the biocompatible composite membrane obtained in step (a), including the semipermeable membrane layer and the nonwoven fabric layer. In step (d), the semipermeable membrane layer is arranged on the outer side, and the nonwoven fabric layer is arranged on the hydrogel side. The order of the above steps is not particularly limited. For example, step (a) can be performed before step (b), between step (b) and step (c), or after step (c). In addition, multiple steps may be performed simultaneously as appropriate.

[0178] The step (a) can be replaced with the following step (A1) or step (A2): Step (A1): A step of forming a biocompatible composite membrane including / consisting of a semipermeable membrane layer formed by laminating a cellulose derivative on a nonwoven fabric used as a support, and a nonwoven fabric layer having a structure in which the cellulose derivative partially penetrates the nonwoven fabric. Step (A2): A step of forming a biocompatible composite membrane by laminating a cellulose derivative on one side of a nonwoven fabric used as a support, and the cellulose derivative partially penetrates the nonwoven fabric.

[0179] <Steps (a), (A1), and (A2)> In step (a), a semipermeable membrane layer formed from a cellulose derivative is prepared using a nonwoven fabric as a support, and a biocompatible composite membrane having a structure in which the cellulose derivative partially penetrates the nonwoven fabric is formed. In step (A1), a biocompatible composite membrane is formed, which includes / consists of a semipermeable membrane layer formed by laminating a cellulose derivative on the nonwoven fabric using the nonwoven fabric as a support, and a nonwoven fabric layer having a structure in which the cellulose derivative partially penetrates the nonwoven fabric. In step (A2), a cellulose derivative is laminated on one side of the nonwoven fabric using the nonwoven fabric as a support, and a biocompatible composite membrane is formed in which the cellulose derivative partially penetrates the nonwoven fabric.

[0180] In step (a), step (A1), or step (A2), a cellulose derivative (e.g., cellulose acetate) is first mixed with a solvent to prepare a solution containing the cellulose derivative (also referred to as a cellulose derivative-containing solution). The solvent to be mixed with the cellulose derivative is not particularly limited, but examples thereof include dimethyl sulfoxide, N,N-dimethylformamide, N,N-dimethylacetamide, dioxane, and acetone.

[0181] In one embodiment of the present invention, the cellulose derivative content in the cellulose derivative-containing solution is about 1 to about 30% by mass, about 5 to about 20% by mass in another embodiment, and about 7 to about 15% by mass in yet another embodiment, based on the total amount of the cellulose derivative-containing solution. The cellulose derivative-containing solution is then brought into contact with a nonwoven fabric to allow the nonwoven fabric to permeate with the cellulose derivative-containing solution. Examples of methods for permeating the nonwoven fabric with the cellulose derivative-containing solution include pouring or applying the cellulose derivative-containing solution onto the surface of the nonwoven fabric, spraying the cellulose derivative-containing solution onto the nonwoven fabric, and immersing the nonwoven fabric in the cellulose derivative-containing solution. For example, the Robb-Srirayan method described in the Examples below can also be used. The nonwoven fabric permeated with the cellulose derivative-containing solution is then immersed in a water bath or the like maintained at about 40 to about 80°C to coagulate the cellulose derivative. This results in a biocompatible composite membrane of one embodiment. Furthermore, this method can also form a biocompatible composite membrane having a structure in which the cellulose derivative partially penetrates the nonwoven fabric layer. The resulting composite membrane can be washed, cut, and stored as needed, preferably in a refrigerator.

[0182] <Step (b)> In step (b), cells or tissues are mixed with a hydrogelatinizable solution (e.g., an alginic acid solution, a gelatinizable solution described in [3-1], [3-2], or [1A-9] to [1A-11] above, etc.). Examples of hydrogelatinizable alginic acid solutions include aqueous sodium alginate solutions and the alginic acid derivatives represented by the aforementioned formulas (H-I), (H-II), (I-A), and (II-A). In step (b), for example, an aqueous solution or physiological saline solution containing about 0.1 to about 5 wt % of the alginic acid derivative is prepared, and the required amount of the cells or tissues is suspended in the solution. Here, the "hydrogelatinizable solution" refers to, for example, two types of solutions: a solution of the alginic acid derivative represented by the aforementioned formula (H-I) and a solution of the alginic acid derivative represented by the aforementioned formula (H-II). Alternatively, for example, two solutions may be used: a solution of the alginic acid derivative represented by the aforementioned formula (IA) and a solution of the alginic acid derivative represented by the aforementioned formula (II-A). In this case, in step (b), these two solutions are prepared separately without being mixed. In this case, the cells or tissues may be mixed in only one of the two solutions, or in both.

[0183] In addition, materials other than alginic acid derivatives, such as the alginic acid derivatives represented by the above-mentioned formulas (HI) and (H-II), and the alginic acid derivatives represented by formulas (IA) and (II-A), may be added to the alginic acid solution that can be hydrogelled. Such materials include, for example, alginic acid (including alginic acid esters and alginates), collagen, hyaluronan, gelatin, fibronectin, elastin, tenascin, laminin, vitronectin, polypeptides, heparan sulfate, chondroitin, chondroitin sulfate, keratan, keratan sulfate, dermatan sulfate, carrageenan, heparin, chitin, chitosan, agarose, agar, cellulose, methylcellulose, carboxymethylcellulose, glycogen and derivatives thereof, as well as fibrin, fibrinogen, thrombin, polyglutamic acid, polylactic acid, polyglycolic acid, lactic acid-glycolic acid copolymer, gellan gum, xanthan gum, galactomannan, guar gum, locust bean gum, and tara gum.

[0184] <Step (c)> In step (c), the solution obtained in step (b) is gelled. For example, when the alginic acid derivatives represented by the aforementioned formulas (H-I) and (H-II) are used as raw materials for the hydrogelable alginic acid solution, or when the alginic acid derivatives represented by formulas (I-A) and (II-A) are used, the two solutions obtained in step (b) are mixed to cause chemical crosslinking via the Huisgen reaction, thereby gelling the solution. Furthermore, by adding or contacting the solution obtained in step (b) with a crosslinking agent such as a solution containing a divalent metal ion (e.g., a calcium chloride aqueous solution, a barium chloride aqueous solution, etc.) or a solution containing a chemical crosslinker, a hydrogel can be produced via bonds (e.g., divalent metal ion bonds, chemical bonds) due to a crosslinking reaction. In step (c), these multiple gelation methods can also be combined. For example, when the alginic acid derivatives represented by the aforementioned formulas (H-I) and (H-II) or the alginic acid derivatives represented by formulas (I-A) and (II-A) are used as raw materials for the hydrogelable alginic acid solution, the two solutions obtained in step (b) can be mixed and then contacted with a solution containing a divalent metal ion, which promotes chemical crosslinking and ionic crosslinking, thereby gelling the solution. This method can also expedite the formation of a hydrogel structure in which cells or tissues are embedded, allowing specific structures to be created instantly. Furthermore, in step (c), the solutions obtained in step (b) can be mixed and a bioprinter can be used to form the structure.

[0185] <Step (d)> In step (d), the cell- or tissue-embedded hydrogel obtained in step (c) is coated with a biocompatible composite membrane comprising a semipermeable membrane layer and a nonwoven fabric layer obtained in step (a), step (A1), or step (A2). The hydrogel is coated with the semipermeable membrane layer of the biocompatible composite membrane facing outward and the nonwoven fabric layer facing the hydrogel (the hydrogel is coated with the surface of the biocompatible composite membrane where the cellulose derivative is laminated on the nonwoven fabric facing outward). As described above, a method for coating the hydrogel with the biocompatible composite membrane includes placing the hydrogel between two biocompatible composite membranes cut to an appropriate size and heat sealing the edges of the two composite membranes. In a preferred embodiment, the hydrogel can be coated with the composite membrane by heat sealing four or fewer times, three or fewer times, two or fewer times, or one time. Through these steps, a biocompatible device according to one embodiment of the present invention can be produced.

[0186] 6-2. Manufacturing Method of Biocompatible Device (2) One embodiment of the manufacturing method includes the following steps (i) to (v): Step (i): A step of preparing a semipermeable membrane layer formed from a cellulose derivative using a nonwoven fabric as a support, thereby forming a biocompatible composite membrane having a structure in which the cellulose derivative partially penetrates the nonwoven fabric. Step (ii): A step of forming an internal space using the biocompatible composite membrane obtained in step (i), in which the semipermeable membrane layer of the biocompatible composite membrane is arranged on the outer side and the nonwoven fabric layer is arranged on the internal space side. Step (iii): A step of mixing cells or tissues (e.g., cells or tissues secreting a biologically active substance) with a hydrogelatinizable solution (e.g., an alginate solution, a gelatinizable solution described in [3-1], [3-2], or [1A-9] to [1A-11] above, etc.). Step (iv): A step of injecting the solution obtained in step (iii) into the internal space formed by the biocompatible composite membrane of step (ii). Step (v): A step of gelling the solution injected in step (iv). The order of the above steps is not particularly limited. For example, steps (i) to (ii) may be performed before step (iii), or may be performed between step (iii) and step (iv). Furthermore, multiple steps may be performed simultaneously as appropriate.

[0187] The step (i) can be replaced with the following step (i-1) or step (i-2): Step (i-1): A step of forming a biocompatible composite membrane including / consisting of a semipermeable membrane layer formed by laminating a cellulose derivative on a nonwoven fabric used as a support, and a nonwoven fabric layer having a structure in which the cellulose derivative partially penetrates the nonwoven fabric. Step (i-2): A step of forming a biocompatible composite membrane by laminating a cellulose derivative on one side of a nonwoven fabric used as a support, and the cellulose derivative partially penetrates the nonwoven fabric.

[0188] <Steps (i), (i-1), and (i-2)> In step (i), a semipermeable membrane layer formed from a cellulose derivative is prepared using a nonwoven fabric as a support, thereby forming a biocompatible composite membrane having a structure in which the cellulose derivative partially penetrates the nonwoven fabric. The specific method for step (i) is the same as step (a) in the biocompatible device manufacturing method (1) described above. In step (i-1), a biocompatible composite membrane is formed, which includes / consists of a semipermeable membrane layer formed by laminating a cellulose derivative on the nonwoven fabric using a nonwoven fabric as a support, and a nonwoven fabric layer having a structure in which the cellulose derivative partially penetrates the nonwoven fabric. The specific method for step (i-1) is the same as step (A1) in the biocompatible device manufacturing method (1) described above. In step (i-2), a cellulose derivative is laminated on one side of the nonwoven fabric using a nonwoven fabric as a support, thereby forming a biocompatible composite membrane in which the cellulose derivative partially penetrates the nonwoven fabric. The specific method of step (i-2) is the same as step (A2) of the above-mentioned method (1) for producing a biocompatible device.

[0189] <Step (ii)> In step (ii), an internal space is formed in the biocompatible composite membrane obtained in step (i), step (i-1), or step (i-2). In this step, the biocompatible composite membrane is arranged so that the semipermeable membrane layer faces outward and the nonwoven fabric layer faces the internal space. Alternatively, the biocompatible composite membrane is arranged so that the surface on which the cellulose derivative is laminated to the nonwoven fabric faces outward. As described above, a method for forming an internal space in the biocompatible composite membrane includes placing two biocompatible composite membranes cut to an appropriate size and heat sealing the edges of the two composite membranes. In a preferred embodiment of the manufacturing method, the internal space can be formed in the composite membrane by heat sealing four or less times, three or less times, two or less times, or one time.

[0190] <Step (iii)> In step (iii), cells or tissues are mixed with a solution capable of hydrogelation. The specific method of step (iii) is the same as step (b) of the above-mentioned method (1) for producing a biocompatible device.

[0191] <Step (iv)> In step (iv), the solution obtained in step (iii) is injected into the internal space formed by the biocompatible composite membrane in step (ii). The method for injecting the solution into the internal space is not particularly limited, and the solution may be injected into the internal space using a pipette or the like in an amount that does not leak from the composite membrane.

[0192] <Step (v)> In step (v), the solution injected in step (iv) is gelled. The specific method for step (v) is the same as step (c) in the biocompatible device manufacturing method (1) described above. By undergoing these steps, the biocompatible device of one aspect of the present invention can be manufactured.

[0193] 7. Physical Properties of Biocompatible Devices The physical properties of biocompatible devices can be evaluated based on the physical properties of the biocompatible composite membrane. <Substance Permeability> The substance permeability of a biocompatible composite membrane according to one embodiment of the present invention can be evaluated using a chamber for a drug oral absorption evaluation system (D / P system). For example, a membrane is placed in a D / P system chamber, and the permeation of each added solution (e.g., insulin solution, glucose solution, albumin solution, dextran solution) is measured over time for up to 24 hours. The substance permeability can then be calculated using the following formula: Permeability (%) = 100 × test substance concentration in permeating solution / test substance concentration at permeation equilibrium. More specifically, substance permeability can be evaluated using the method described in the Examples below.

[0194] In one embodiment of the biocompatible composite membrane of the present invention, the glucose permeability after 24 hours, as measured by the method described in the Examples below, is about 80% or more, in another embodiment about 85% or more, and in yet another embodiment about 90% or more. In another embodiment of the biocompatible composite membrane of the present invention, the insulin permeability after 24 hours, as measured by the method described below, is about 30% or more, in another embodiment about 40% or more, and in yet another embodiment about 50% or more. In another embodiment of the biocompatible composite membrane of the present invention, the dextran (molecular weight: about 150 kD) permeability after 24 hours, as measured by the method described below, is about 30% or less, in another embodiment about 20% or less, and in yet another embodiment about 10% or less.

[0195] <Strength> The strength of the biocompatible composite membrane of one embodiment of the present invention can be measured by a tensile test, a tear test, a burst test, or the like. For example, it can be measured by a tensile test in accordance with JIS K 7127 or ASTM D882. In one embodiment of the biocompatible device of the present invention, the strength of the composite membrane measured by the tensile test is, for example, 1 MPa or more, preferably about 1 to about 100 MPa, and more preferably about 5 to about 100 MPa. Furthermore, the tear test and the burst test can be measured by common test methods.

[0196] The heat-seal integrity of the biocompatible composite membrane of one embodiment of the present invention can be measured by the method described in the Examples below. Alternatively, the heat-seal integrity can be measured by a method adapted from a filter integrity test. For example, the bubble point value can be measured using an integrity test kit for small-volume devices (Merck, SLTEST000) and used as an index of heat-seal integrity.

[0197] 8. Biocompatibility of Devices and Composite Membranes The devices and composite membranes provided herein are biocompatible. The alginic acid derivatives and crosslinked alginic acid constituting the hydrogels of the present invention have been shown to have good biocompatibility, similar to alginic acid (see, for example, PCT / JP2019 / 023478, PCT / JP2020 / 047100, and PCT / JP2019 / 007655).

[0198] The biocompatibility of the device and composite membrane herein will be confirmed in the examples relating to biocompatibility described later.

[0199] 9. Uses and Methods of Use of Biocompatible Devices The biocompatible devices provided herein can be used as bioimplantation devices. A certain embodiment of the biocompatible device can be suitably used for the treatment of diabetes. The device can be used to treat type 1 diabetes, type 2 diabetes, and diabetes caused by other mechanisms, and is preferably used for the treatment of type 1 diabetes. A preferred embodiment of the device not only has excellent biocompatibility, but also excellent stability, low cytotoxicity, little adhesion or inflammation at the implantation site, little gel dissolution, and is capable of maintaining a blood glucose lowering effect and regulating blood glucose over a long period of time.

[0200] The implantation site of the device is not particularly limited, and examples thereof include subcutaneous, intramuscular, intraperitoneal, intrahepatic, intraomental, and subrenal capsule sites, but implantation is preferably subcutaneous, intramuscular, intraperitoneal, or intraomental. The device can be implanted into a living body by incision and placement, endoscopy, laparoscopy, or the like.

[0201] 10. Biological Transplant Kit A biological transplant kit is provided herein. In some embodiments, the biological transplant kit includes a biocompatible device having an internal space formed by a biocompatible composite membrane including a semipermeable membrane layer and a nonwoven fabric layer, and a container containing cells or tissue that secrete a biologically active substance, and is used so that the cells or tissue are encapsulated in the internal space. The terms "biocompatible device," "cells or tissue," etc. are as explained above.

[0202] A biological transplant kit according to one embodiment may include, in a single package, (1) a pack containing a biocompatible device and (2) a container (e.g., a vial) containing frozen cells or tissues secreting a biologically active substance. The biocompatible device (1) may be packaged with a portion of the biocompatible composite membrane (e.g., one of the four sides in the case of a flat plate) unfused so that the cells or tissues can be sealed in the internal space before use. In this case, the cells or tissues can be sealed in the internal space of the biocompatible device before use, and the unfused portion (e.g., the remaining side in the case of a flat plate) can be heat-sealed to seal the internal space.

[0203] In one embodiment, the biological transplant kit may further include, in addition to the above (1) and (2), (3) a container such as a vial containing a freeze-dried alginic acid derivative that is gelled by chemical crosslinking, and (4) a container such as a vial containing physiological saline for dissolution, all in one package. In another embodiment, the biological transplant kit may contain the cells or tissue of (2) above and the alginic acid derivative of (3) above, dissolved in a container such as a vial in advance. In yet another embodiment, the biological transplant kit may contain the cells or tissue of (2) above mixed with a solution of the alginic acid derivative of (3) above, gelled, and then sealed in a pack in advance. The kit of the present invention may further include accessories such as syringes, pipettes, injection needles, dedicated fillers, and instruction manuals, as appropriate.

[0204] All documents and publications mentioned herein are hereby incorporated by reference in their entirety for any purpose.

[0205] Furthermore, the objects, features, advantages, and ideas of the present invention will be apparent to those skilled in the art from the description in this specification, and those skilled in the art will be able to easily implement the present invention from the description in this specification. The best mode for carrying out the invention and specific examples show preferred embodiments of the present invention and are shown for the purpose of illustration and explanation, and are not intended to limit the present invention thereto. It will be apparent to those skilled in the art that various modifications can be made based on the description in this specification within the spirit and scope of the present invention disclosed in this specification.

[0206] The chemically modified alginic acid derivatives used in the following examples were synthesized in accordance with the method described in the examples of WO 2020 / 262642. The alginic acid derivative (A01) or (A02) having a cyclic alkyne group, and the alginic acid derivative (N01) having an azide group were used. By mixing these two alginic acid derivatives, a Huisgen reaction occurs, and chemically crosslinked alginic acid containing the crosslinked structures of the formulas (LZ-8) and (LZ-8-r), or chemically crosslinked alginic acid containing the crosslinked structures of the formulas (LY-2) and (LY-2-r), is formed, forming a hydrogel. In addition, by using an ionic crosslinking reaction with a divalent metal ion in combination, the formation of the hydrogel structure can be accelerated, and in some examples, it is used in combination.

[0207] [Example A] Example of composite membrane production (1) Preparation of membrane-forming solution (dope) Cellulose acetate (LT-75) (manufactured by Daicel Corporation) 11 wt%, PEG200 (manufactured by Hayashi Pure Chemical Industries) 10 wt%, and dimethyl sulfoxide (DMSO) (manufactured by Nacalai Tesque) 79 wt% were weighed to prepare a mixed solution of PEG200 and DMSO. Next, cellulose acetate was gradually added to the mixed solution while stirring at 70 to 80°C to obtain a dope for membrane formation.

[0208] (2) Preparation of Composite Membranes. Composite membranes were prepared using the Loeb-Sourirajan method, a method for producing asymmetric porous membranes (see, for example, S. Loeb and S. Sourirajan, ibid. No. 59-28 (1959)). A nonwoven fabric (HOP-30H) (manufactured by Hirose Paper Co., Ltd.) was cut to the same size as a glass plate and attached to the glass plate with tape. A film applicator was placed on one end of the nonwoven fabric on the glass plate, and dope kept at 60°C was evenly dispensed from one end of the applicator to a width of approximately 1 cm. The applicator was pressed against the nonwoven fabric and slid to cast the dope onto the nonwoven fabric. The nonwoven fabric and the glass plate were then submerged in a 60°C water bath for several minutes to solidify the dope, yielding a composite membrane in which cellulose acetate was laminated to the nonwoven fabric. The obtained composite membrane was washed together with the glass plate with water, and then the composite membrane was peeled off from the glass plate and immersed in water overnight to thoroughly remove the solvent contained in the membrane.

[0209] (3) Properties, Physical Properties, etc. of the Composite Membrane - Heat-sealing integrity: Two composite membranes cut into rectangles measuring approximately 3.5 cm x 2.0 cm were stacked with the nonwoven fabric layers facing each other, and three sides, excluding one 2.0 cm side, were heat-sealed using a polysealer (Fuji Impulse, P-300) to form an envelope-like structure. A coloring liquid (5 mg / mL TRITC-dextran 150 solution (TdB Labs) prepared in saline) was injected into the internal space created by the heat-sealing, and the remaining side was heat-sealed. After a certain period of time, it was confirmed that the coloring liquid did not seep outside the sealed area. If there was no liquid seepage, it was determined that the heat-sealing properties were intact. The composite membrane of the present invention did not seep outside the sealed area, confirming sufficient heat-sealing properties. - Strength: Visual and tactile observations confirmed that the strength of the composite membrane was higher than that of commercially available membranes.

[0210] Example B: Evaluation of Permeability of Composite Membranes. The permeability of composite membranes was evaluated using a horizontal chamber (Osaka Rika) for the drug oral absorption evaluation system (D / P system). A 3 cm x 3 cm sample of the membrane was immersed in a 0.1% Tween 20 solution (dialysis solution) prepared with saline and then placed in the chamber. 9 mL of dialysis solution was placed in the chamber's addition side, and 7.5 mL in the chamber on the opposite side (recovery side) separated by the membrane. Then, with both chambers agitated (200 rpm) at room temperature, 1 mL of the following test solution was added to the addition side to initiate the permeability test. The test was initiated upon addition of each solution, and samples were taken from the recovery side 2 and 24 hours after the start of the test. After 24 hours, samples were also taken from the addition side. The test substance concentration in each sampled solution was calculated based on the quantitative value obtained using a measurement kit for each test substance or the quantitative value obtained by fluorescent measurement of the labeled substance. The permeability was then calculated using the following formula: (Membranes used) Example: Composite membrane prepared in Example A Reference example: Dialysis tube "Spectra / Pore CE (molecular weight cutoff: 100,000)" manufactured by Repligen (hereinafter referred to as commercially available membrane) (Test solutions) 100 mg / dL glucose solution (manufactured by Otsuka Pharmaceutical) 2000 ng / mL insulin solution (manufactured by Eli Lilly) 500 μg / mL FITC-albumin solution (manufactured by Sigma) 2000 μg / mL TRITC-dextran150 solution (manufactured by TdB Labs) (Formula for calculating substance permeability) Permeability (%) = 100 × concentration of test substance in sampled solution / concentration of test substance at permeability equilibrium

[0211] The substance permeabilities of the composite membranes of the examples and the commercially available membranes are shown in Table 16. The orientation of the evaluation membranes is shown here with the semipermeable membrane layer side as the outside and the nonwoven fabric layer side as the inside for the composite membrane and the outside of the cylindrical dialysis tube as the outside and the inside as the inside for the commercially available membrane, respectively, and the direction of substance permeation in the evaluation system is indicated by an arrow. For example, "composite membrane (outside → inside)" means that the evaluation was performed with the semipermeable membrane layer side of the composite membrane facing the addition side and the nonwoven fabric layer side facing the recovery side. As shown in Table 16, the composite membranes of the examples exhibited substance permeability equivalent to that of commercially available membranes, and it was confirmed that they functioned as semipermeable membranes.

[0212] [Example C] Evaluation of Complement Sequestration Ability The device containing a solution of sensitized sheep red blood cells was shaken in a human serum-mixed buffer solution, and the hemolysis rate was measured to evaluate the device's complement sequestration ability. (1) Preparation of a Device for Evaluating Complement Sequestration Ability The above alginate derivatives (A02) and (N01) were dissolved in CH50 buffer (the buffer solution provided with One Point CH50 "Seiken" (Denka Co., Ltd.)) to a final concentration of 0.5% alginate derivative (equal mixture of alginate derivative (A02) and alginate derivative (N01) at a final concentration of 0.25%), and one-third of the solution was added with sensitized sheep red blood cells (red blood cell-containing alginate solution). (Preparation of Hydrogel) A commercially available membrane (dialysis tubing with a flat width of 1 cm) was cut to a length of approximately 2.7 cm, and one 1 cm wide edge was heat-pressed to form an envelope-like structure. 100 μL of red blood cell-containing alginate solution was injected into the internal space created by molding. After the remaining side was heat-pressed, the device was immersed in a 10 mmol / L BaCl2 solution for 10 minutes to promote hydrogel formation. (Example Device) The hydrogel prepared above was removed from the bag. Two rectangular sheets of the composite membrane prepared in Example A, each approximately 1.5 cm x 2.5 cm in size, were prepared and placed on top and bottom of the gel with the nonwoven fabric layer facing the gel side. The four sides were heat-pressed to form an example device. (Reference Example Device) The hydrogel wrapped in the commercially available membrane prepared above was used as a reference example device.

[0213] (2) Evaluation Method: Evaluation devices containing the composite membrane of the example or a commercially available membrane encapsulating hydrogel were immersed in 3 mL of CH50 buffer containing 10 μL of human serum (complement control "Seiken" HC(H) (Denka Corporation)) and incubated at 30°C for 24 hours. After incubation, the absorbance at 541 nm of each supernatant was measured, and the hemolysis rate was calculated. As a control, a mixture of CH50 buffer and sensitized sheep red blood cells at a volume ratio of 3:1 (red blood cell solution) was used instead of the device. 100 μL of the red blood cell solution was added to 3 mL of CH50 buffer containing 10 μL of human serum and incubated in the same manner as the device. This was evaluated as a complement hemolysis control group. The hemolysis rate of each sample was calculated by considering 100 μL of the red blood cell solution added to 3.01 mL of water as 100% hemolysis.

[0214] As a result, the hemolysis rate in the complement hemolysis control group was about 70%, but in both the Example device using the composite membrane of the Example and the Reference Example device using the commercially available membrane, the hemolysis rate was 1% or less, and almost no hemolysis was observed. Therefore, it was confirmed that the device made using the composite membrane of the Example has the ability to sequester complement.

[0215] Example D: Glucose-responsive insulin secretion test The glucose-responsive insulin secretion ability was evaluated using a device containing insulin-secreting cells. (1) Device preparation: MIN6 cells (2.5 × 10 cells) derived from pancreatic β cells were added to 100 μL of a solution prepared by mixing equal amounts of 0.5% aqueous solutions of the alginic acid derivatives (A01) and (N01). 6 Two composite membranes prepared in Example A were cut into rectangles measuring approximately 2 cm x 1 cm. The two were stacked with the nonwoven fabric layers facing each other, and three sides, excluding one 1 cm side, were heat-pressed to form an envelope-like structure. Similarly to Example C, a commercially available membrane was also molded into an envelope-like structure of the same size. The cell suspension was then injected into each internal space created by molding. After the remaining side of each structure was heat-pressed, it was immersed in a 50 mmol / L CaCl2 solution for 10 minutes to promote hydrogel formation, and used as an example device and a reference example device for evaluation.

[0216] (2) Evaluation method After culturing the device at 37°C for one day using a culture medium (Optimized DMEM (AddexBio) containing 15% FBS), the glucose concentration-dependent insulin secretion was evaluated. Specifically, the device was immersed in a 2 mmol / L glucose solution for one hour or a 20 mmol / L glucose solution for two hours, and the insulin concentration secreted into the solution under each condition was measured. This allows the insulin secretion ability of the cells in response to glucose concentration to be evaluated.

[0217] The results of the glucose-responsive insulin secretion test for the Example device prepared using the composite membrane of the Example or the Reference Example device prepared using the commercially available membrane are shown in Table 17. In Table 17, "HG" represents high glucose (20 mmol / L glucose solution) and "LG" represents low glucose (2 mmol / L glucose solution). The device fabricated using the composite membrane of the present example showed glucose-responsive insulin secretion equivalent to that of the device using the commercially available membrane, suggesting that the two types of devices exert equivalent cellular functions.

[0218] [Example E] Biocompatibility test of composite membranes (rats) (1) Preparation of membranes The following four types of membranes were prepared for implantation: (Example): The composite membrane prepared in Example A was sterilized in advance by UV irradiation, and cut into a rectangle measuring approximately 2.8 cm x approximately 1.8 cm. Two of these were stacked with the nonwoven fabric layers facing each other, and the four sides were heat-pressed together. (Reference Example): Two commercially available membranes were cut into a rectangle measuring approximately 2 cm x approximately 1 cm, and the four sides were heat-pressed together. (Adhesion-inducing control): A membrane made by cutting a polyurethane containing 0.75% zinc diethyldithiocarbamate (Hadano Research Institute, Food and Drug Safety Center, General Incorporated Foundation) into a piece approximately 2 cm x approximately 1 cm and sterilizing it in an autoclave. (Non-effect control): A membrane made by cutting a high-density polyethylene sheet (Hadano Research Institute, Food and Drug Safety Center, General Incorporated Foundation) into a piece approximately 2 cm x approximately 1 cm and sterilizing it in an autoclave.

[0219] (2) Implantation and Evaluation Methods The above four membranes were implanted intraperitoneally into 8-week-old Sprague-Dawley rats (Japan SLC) under anesthesia, and the wounds were closed. The rats were then raised and observed. Two weeks after implantation, the abdomen was opened under anesthesia, and intraperitoneal adhesions and the state of the implants were visually observed. The visual evaluation results of membrane adhesions, membrane changes, and inter-organ adhesions are shown in Table 18.

[0220] As shown in Table 18, the composite membrane of the Example did not show any membrane adhesions, membrane changes, or inter-organ adhesions, making it suitable for transplantation. On the other hand, slight inter-organ adhesions were observed in the commercially available membrane of the Reference Example. Weight loss was observed in all cases, but this was transient and likely due to the effects of treatment, and the animals subsequently recovered. No abnormalities were observed in their general condition.

[0221] [Example F] Device Biocompatibility Test (Dog, Miniature Pig) (1) Device Fabrication The following two types of implantable devices were fabricated. (Example Device) Two composite membranes fabricated in Example A, cut into squares measuring approximately 2.4 cm x 2.4 cm, were stacked with the nonwoven fabric layers facing inward, and three sides were thermocompressed to form an envelope-like structure. 156 μL of a mixture of equal amounts of 1% aqueous solutions of alginic acid derivatives (A02) and (N01) was injected into the internal space created by the molding. After thermocompression of the remaining side, the structure was immersed in a 10 mmol / L BaCl2 solution for 10 minutes to promote hydrogel formation, resulting in an example device. (Reference Example Device) Two commercially available membranes cut into squares measuring approximately 2.4 cm x 2.4 cm were stacked, and three sides were thermocompressed to form an envelope-like structure. A 156 μL mixture of equal amounts of 1% aqueous solutions of the alginic acid derivatives (A02) and (N01) was poured into the internal space created by molding. After the remaining side was thermocompressed, the structure was immersed in a 10 mmol / L BaCl2 solution for 10 minutes to promote the formation of a hydrogel, resulting in a reference device.

[0222] (2) Implantation and evaluation methods Each device was implanted into the abdominal cavity of beagle dogs and minipigs (Goettingen minipigs) under anesthesia, and the wounds were closed. The animals were then raised and observed. Four weeks after implantation, the abdomen was opened under anesthesia, and intraperitoneal adhesions and the state of the implant were visually observed.

[0223] As a result, slight adhesion with surrounding tissues was observed in the cases where the Reference Example device was implanted. On the other hand, no adhesion, inflammation, etc. was observed in the cases where the Example device was implanted. Furthermore, no effect on abdominal organs was observed in the cases where the Example device was implanted.

[0224] [Example G] Fabrication of devices using a 3D printer and biocompatibility tests (dogs and minipigs) (1) Fabrication of devices An aqueous solution containing alginic acid derivatives (A02) and (N01) (final concentration of each: 0.5%), alginic acid (A-2: sodium alginate manufactured by Mochida Pharmaceutical Co., Ltd., listed in Table 1) (final concentration: 1%), and BaCl2 (final concentration: 2.5 mmol / L) was prepared and printed on a 3D printer (BIO X TM Using a 410 μm diameter nozzle (manufactured by CELLINK), the aqueous solution was injected into a rectangular shape of approximately 6.5 cm x 3 cm. A 10 mmol / L BaCl2 solution was then layered over the injected solution and allowed to stand for 10 minutes to form a hydrogel. Two rectangular sheets of the composite membrane prepared in Example A, cut into approximately 7 cm x 3.5 cm rectangles, were placed on top and bottom of the hydrogel, with the nonwoven fabric layer facing the gel side, and the four sides were thermocompressed to form an implantable device.

[0225] (2) Implantation Method and Evaluation Method The fabricated devices were implanted in beagle dogs and minipigs (Goettingen minipigs) in the same manner as in Example F (device biocompatibility test (dogs, minipigs)) and evaluated.

[0226] As a result, no adhesions or inflammation thought to be caused by the device materials were observed in the device fabricated using the hydrogel fabricated with a 3D printer and the composite membrane fabricated in Example A.

[0227] Example H: Implantation Site Evaluation in Biocompatibility Tests (Dogs and Minipigs) (1) Device Fabrication Two composite membranes prepared in Example A were cut into rectangles measuring approximately 7 cm x 3.5 cm. Two sheets were stacked with the nonwoven fabric layers facing each other, and the three edges, excluding one short edge, were thermocompressed to form an envelope-like structure. 700 μL of an equal mixture of 1% aqueous solutions of the alginic acid derivatives (A02) and (N01) was injected into the resulting internal space. After thermocompression of the remaining edge, the structure was immersed in a 10 mmol / L BaCl2 solution for 10 minutes to promote hydrogel formation and prepare a device for implantation.

[0228] (2) Implantation method and evaluation method According to the method of Example F (Device biocompatibility test (dogs, minipigs)), beagle dogs and minipigs (Goettingen minipigs) were used, and the above device was implanted into the following three implantation sites. Implantation site 1: Intraperitoneal cavity Under anesthesia, the device was implanted into the abdominal cavity and the incision was closed. Implantation site 2: Intraomental pocket After laparotomy under anesthesia, the omentum was folded in half and sewn to create a pocket. The device was implanted in the pocket and the incision was closed. Implantation site 3: Abdominal wall After laparotomy under anesthesia, the device was sutured to the abdominal wall off the midline and the incision was closed.

[0229] Four weeks after implantation, the patient underwent laparotomy under anesthesia, and the condition of the implant and intraperitoneal adhesions were visually examined. As a result, no adhesions or inflammation thought to be due to the device material were observed, regardless of the implantation site.

[0230] Example I: Evaluation of the efficacy of the device (in mice) (1) Preparation of the device MIN6 cells (2.2 × 10 cells) were cultured in the same manner as in Example D in 50 μL of a solution prepared by mixing equal amounts of 0.5% aqueous solutions of the alginic acid derivatives (A01) and (N01). 6Two composite membranes prepared in Example A were cut into rectangles measuring approximately 2 cm x 1 cm. The two were stacked with the nonwoven fabric layers facing each other, and three sides, excluding one 1 cm side, were heat-pressed to form an envelope-like structure. Similarly to Example C, a commercially available membrane was also molded into an envelope-like structure of the same size. The cell suspension was injected into each internal space created by molding. After the remaining side of each structure was heat-pressed, it was immersed in a 50 mmol / L CaCl2 solution for 10 minutes to promote hydrogel formation, and these were used to evaluate the Example and Reference Example devices.

[0231] (2) Implantation and evaluation methods: Each device was implanted intraperitoneally into C57BL / 6 mice that had been induced with diabetes by streptozotocin under anesthesia, and the wound was closed. The mice were then kept and observed. Blood glucose levels were measured over a 4-week period after implantation.

[0232] The efficacy of the example device and the reference example device is shown in Table 5. Here, a cure was judged when the blood glucose level at each measurement decreased by 200 mg / dL or more compared to the blood glucose level before implantation. The cure was judged based on the blood glucose levels of each animal 5 and 25 days after implantation, and the proportion of cured animals out of all animals was calculated as the cure rate.

[0233] As a result, the Example device was confirmed to be as effective as or better than the Reference Example device using a commercially available membrane. In particular, the healing rate 25 days after implantation was 33% for the Reference Example device, while the Example device showed a higher healing rate of 80%. The unevenness of the nonwoven fabric surface may have functioned as an anchor for the hydrogel, preventing the gel from shifting, folding, or collapsing within the device. As a result, it is thought that the utilization rate of the cells embedded in the hydrogel was improved, further enhancing the therapeutic effect.

[0234] REFERENCE SIGNS LIST 1 Biocompatible device 2 Biocompatible composite membrane 21 Nonwoven fabric layer (nonwoven fabric) 211 Portion not penetrated by semipermeable membrane components 212 Portion in which semipermeable membrane components have partially penetrated into nonwoven fabric 22 Semipermeable membrane layer (semipermeable membrane) 3 Internal space 4 Cells or tissues 5 Hydrogel 6 Fused portion

[0235] The present invention may include the following aspects: [1] A biocompatible device having an internal space capable of containing cells or tissues secreting a biologically active substance, the internal space being formed by placing a biocompatible composite membrane comprising a semipermeable membrane layer and a nonwoven fabric layer on the device, with the semipermeable membrane layer on the outer side and the nonwoven fabric layer on the internal space side, the semipermeable membrane layer comprising a cellulose derivative laminated on the nonwoven fabric using the nonwoven fabric as a support, the nonwoven fabric layer comprising a thermoplastic nonwoven fabric. [2] The device according to [1] above, which has immunoisolation ability. [3] The device according to [1] above, in which the cellulose derivative is cellulose acetate. [4] The device according to [1] above, in which the nonwoven fabric comprises one or more resins selected from the group consisting of polyethylene, polypropylene, and polyethylene terephthalate. [5] The device according to [1] above, in which the internal space is formed by partial fusion of the nonwoven fabric surface of the composite membrane that is not laminated with the cellulose derivative. [6] The device according to [1], wherein the composite membrane has a structure in which the cellulose derivative partially penetrates into a nonwoven fabric. [7] The device according to [1], wherein the cells secreting a bioactive substance are insulin-secreting cells or pancreatic islets. [8] The device according to [1], wherein the cells or tissue secreting a bioactive substance are encapsulated in the internal space using a hydrogel as a carrier. [9] The device according to [1], wherein the thickness of the composite membrane is 10 to 1500 μm.

[10] The device according to any one of [1] to [9], which is used for biological transplantation.

[11] A biocompatible composite membrane for producing the device according to any one of [1] to [9].

[12] A biological transplantation kit comprising a biocompatible device in which an internal space is formed by a biocompatible composite membrane including a semipermeable membrane layer and a nonwoven fabric layer, and a container containing cells or tissue secreting a bioactive substance, wherein the cells or tissue are encapsulated in the internal space.

[13] A method for producing a biocompatible device, comprising a step of covering a hydrogel in which cells or tissues that secrete a biologically active substance are embedded with a biocompatible composite membrane including a semipermeable membrane layer and a nonwoven fabric layer, wherein in the step, the semipermeable membrane layer is positioned on the outside and the nonwoven fabric layer is positioned on the hydrogel side.

[14] A method for producing a biocompatible device, comprising the steps of injecting a solution containing cells or tissues that secrete a biologically active substance, the cells or tissues being mixed with a hydrogelatinizable solution, into an internal space formed by a biocompatible composite membrane comprising a semipermeable membrane layer and a nonwoven fabric layer, and gelling the hydrogelatinizable solution, wherein in the step, the semipermeable membrane layer of the composite membrane is on the outer side and the nonwoven fabric layer is on the internal space side.

Claims

1. A biocompatible device having an internal space capable of containing cells or tissues that secrete a biologically active substance, the internal space being formed by placing a biocompatible composite membrane comprising a semipermeable membrane layer and a nonwoven fabric layer, the semipermeable membrane layer being on the outer side of the device and the nonwoven fabric layer being on the internal space side, the semipermeable membrane layer comprising a cellulose derivative laminated on the nonwoven fabric using the nonwoven fabric as a support, and the nonwoven fabric layer comprising a nonwoven fabric having thermoplastic properties.

2. The device of claim 1, having immunoisolation capabilities.

3. The device of claim 1, wherein the cellulose derivative is cellulose acetate.

4. The device of claim 1, wherein the nonwoven fabric comprises one or more resins selected from the group consisting of polyethylene, polypropylene, and polyethylene terephthalate.

5. The device according to claim 1, wherein the internal space is formed by partial fusion of the nonwoven fabric surface of the composite membrane that is not laminated with a cellulose derivative.

6. The device of claim 1, wherein the cells secreting the bioactive substance are insulin-secreting cells or pancreatic islets.

7. The device according to claim 1, wherein the cells or tissues secreting the biologically active substance are encapsulated in the internal space using a hydrogel as a carrier.

8. The device of claim 1, wherein the thickness of the composite membrane is between 10 and 1500 μm.

9. A device according to any one of claims 1 to 8, for use in biological implantation.

10. A biocompatible composite membrane for producing a device according to any one of claims 1 to 8.

11. A kit for biological transplantation comprising: a biocompatible device in which an internal space is formed by a biocompatible composite membrane including a semipermeable membrane layer and a nonwoven fabric layer; and a container containing cells or tissue that secrete a biologically active substance, the cells or tissue being used so as to be enclosed in the internal space.

12. A method for producing a biocompatible device, comprising the step of covering a hydrogel in which cells or tissues that secrete a biologically active substance are embedded with a biocompatible composite membrane comprising a semipermeable membrane layer and a nonwoven fabric layer, in which the semipermeable membrane layer is placed on the outside and the nonwoven fabric layer is placed on the hydrogel side.

13. A method for producing a biocompatible device, comprising the steps of injecting a solution containing cells or tissues secreting a biologically active substance, the cells or tissues being mixed with a hydrogellable solution, into an internal space formed by a biocompatible composite membrane comprising a semipermeable membrane layer and a nonwoven fabric layer, and gelling the hydrogellable solution, wherein in the step, the semipermeable membrane layer of the composite membrane is on the outer side and the nonwoven fabric layer is on the internal space side.

Images