Macroencapsulated devices including immunomodulatory compounds

Abstract

Large encapsulated devices and methods of using the same are disclosed. In one embodiment, a macroencapsulated device may include one or more membrane layers configured to encapsulate a population of cells and at least one immunomodulatory compound. The immunomodulatory compounds may be included in an immunomodulatory material, including, for example, a matrix. In some cases, the immunomodulatory material or compound may mitigate undesirable foreign body responses during implantation of the device, is low in cytotoxicity, and / or is capable of promoting cell viability.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority under 35 U.S.C. §119(e) to U.S. Provisional Application Serial No. 63 / 445,517, filed on February 14, 2023, which is incorporated by reference in its entirety for all purposes. Technical Field

[0003] The disclosed embodiments relate to macroencapsulated devices that include immunomodulatory compounds. Background Art

[0004] Therapeutic devices that deliver biological products can be used to treat various metabolic disorders, such as diabetes. Some of these devices include large encapsulated devices that can be used to house cells capable of producing a desired biological product, such as insulin. The devices can be implanted at a location in a subject to provide the desired biological product. Summary of the Invention

[0005] In some aspects, a macroencapsulated device is provided. In some embodiments, a macroencapsulated device comprises: a first membrane layer; a second membrane layer disposed on the first membrane layer, wherein the first membrane layer and the second membrane layer are bonded together to form a seal that extends at least partially around an interior volume disposed between the first membrane layer and the second membrane layer, wherein the first membrane layer and / or the second membrane layer comprise pores; and at least one immunomodulatory compound associated with the first membrane layer and / or the second membrane layer, wherein the at least one immunomodulatory compound comprises at least one selected from the group consisting of a toll-like receptor 4 (TLR4) inhibitor, a nuclear factor kappa light-chain enhancer of activated B cells (NF-κB) inhibitor, a colony stimulating factor 1 receptor (CSF1R) inhibitor, and a NOD, LRR, and pyrin domain-containing protein 3 (NLRP3) inflammasome inhibitor.

[0006] In some aspects, a method for delivering a therapeutic composition produced by a cell population is provided. In some embodiments, the method comprises: diffusing the therapeutic composition across at least one membrane layer that at least partially encapsulates the cell population; and exposing the tissue surrounding the at least one membrane layer to at least one immunomodulatory compound selected from the group consisting of a toll-like receptor 4 (TLR4) inhibitor, a nuclear factor kappa light-chain enhancer of activated B cells (NF-κB) inhibitor, a colony-stimulating factor 1 receptor (CSF1R) inhibitor, and a NOD, LRR, and pyrin domain-containing protein 3 (NLRP3) inflammasome inhibitor.

[0007] In some aspects, an immunomodulatory material is provided. In some embodiments, the immunomodulatory material comprises: a matrix; and at least one immunomodulatory compound included in the matrix, wherein the at least one immunomodulatory compound is selected from the following group: a toll-like receptor 4 (TLR4) inhibitor, a nuclear factor kappa light-chain enhancer of activated B cells (NF-κB) inhibitor, a colony-stimulating factor 1 receptor (CSF1R) inhibitor, and a NOD, LRR, and pyrin domain-containing protein 3 (NLRP3) inflammasome inhibitor.

[0008] In some aspects, a composition is provided. In some embodiments, the composition comprises: a cell population; and at least one immunomodulatory compound selected from the group consisting of a toll-like receptor 4 (TLR4) inhibitor, a nuclear factor kappa light-chain enhancer of activated B cells (NF-κB) inhibitor, a colony-stimulating factor 1 receptor (CSF1R) inhibitor, and a NOD, LRR, and pyrin domain-containing protein 3 (NLRP3) inflammasome inhibitor.

[0009] It should be understood that the aforementioned concepts and the additional concepts discussed below can be arranged in any suitable combination, as the present disclosure is not limited in this respect. In addition, other advantages and novel features of the present disclosure will become apparent from the following detailed description of various non-limiting embodiments when considered in conjunction with the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] The accompanying drawings are not intended to be drawn to scale. In the drawings, each identical or nearly identical component shown in various figures may be represented by a like numeral. For clarity, not every component may be labeled in every drawing. In the drawings:

[0011] Figure 1A shows a side view of a macroencapsulation device prior to loading according to one embodiment;

[0012] Figure 1B According to one embodiment Figure 1A A top view of a macroencapsulation device;

[0013] Figure 1C According to one embodiment, after loading with the desired material Figure 1A A side view of a macroencapsulation device;

[0014] Figure 2 shows a perspective view in cross-section of a portion of a first exemplary macroencapsulated device after being filled with a desired material according to one embodiment;

[0015] Figure 3A flow chart illustrating a method for delivering a therapeutic composition produced by a cell population contained within a macroencapsulated device, according to one embodiment;

[0016] Figure 4 A schematic diagram illustrating an immunomodulatory material associated with a macroencapsulated device according to one embodiment is shown;

[0017] Figure 5 is a graph showing the ratio of IL-10 to IL-1β secreted by M0 macrophages in the presence of a panel of compounds according to one embodiment;

[0018] Figure 6 is a graph showing the amount of IL-1β per cell secreted by M0 macrophages in the presence of a panel of compounds according to one embodiment;

[0019] Figure 7 is a graph showing the amount of IL-1β per cell secreted by M0 macrophages in the presence of MCC950 according to one embodiment;

[0020] Figures 8A-8D Tannic acid ( Figure 8A ), pexidartinib ( Figure 8B )、MCC950( Figure 8C ) and TAK242( Figure 8D ) schematic diagram of the molecular structure;

[0021] Figures 9A-9B is a diagram showing the composition of M0 macrophages ( Figure 9A ) or M1 macrophages ( Figure 9B ) Graph showing the fold change in secreted IL-1β under each compound condition relative to the control (DMSO control without any compound);

[0022] Figure 10 is a graph showing the fold change in cell viability of rat pancreatic islets in the presence of a panel of immunomodulatory compounds according to one embodiment;

[0023] Figure 11 is a graph showing fluctuations in cell number when cells are cultured in the presence of various compounds according to one embodiment;

[0024] Figures 12A-12B is a graph showing the presence of an EGM-2 control ( Figure 12A ) and EGM-2 with DMSO ( Figure 12B ) Microscopic images of endothelial network formation by HUVECs;

[0025] Figures 12C-12E is a microscopy image showing endothelial network formation of HUVECs in the presence of various concentrations of tannic acid according to one embodiment;

[0026] Figures 12F-12H is a microscopy image showing endothelial network formation by HUVECs in the presence of various concentrations of pecidinib according to one embodiment;

[0027] Figure 12I-11 2L is a microscopy image showing endothelial network formation by HUVECs in the presence of various concentrations of TAK242 according to one embodiment;

[0028] Figures 12M-12Q is a microscopy image showing endothelial network formation by HUVECs in the presence of various concentrations of MCC9550 according to one embodiment;

[0029] Figure 12R-12T is a microscopy image showing endothelial network formation by HUVECs in the presence of various concentrations of GDC-2394 according to one embodiment;

[0030] Figures 13A-13B According to one embodiment, the lyophilized fibrin glue containing the immunomodulatory compound is prepared before swelling with PBS ( Figure 13A ) and after swelling ( Figure 13B )

[0031] Figures 14A-14B According to one embodiment, the Figure 14A ) and 100% fibrinogen ( Figure 14B ) SEM image of fibrin glue;

[0032] Figures 15A-15B is a graph showing the extraction of tannic acid from a sample containing 100% fibrinogen ( Figure 15A ) and 50% fibrinogen ( Figure 15B ) is a graph showing the profile of fibrin glue release;

[0033] Figures 16A-16C is a diagram showing the MCC950 from fibrin glue ( Figure 16A ), TAK242 from fibrin glue ( Figure 16B ) and Pecidinib from fibrin glue ( Figure 16C ) a graph of the profile of the release;

[0034] Figure 16D is a graph showing the stability of MCC950, Pesidatinib, and TAK242 in aqueous media according to one embodiment;

[0035] Figures 17A-17B Fibrin glue containing immunomodulatory drugs is shown to be in indirect contact with macrophages according to one embodiment ( Figure 17A ) and direct contact ( Figure 17B ) a cross-sectional view of a device;

[0036] Figure 18 is a graph showing the fold change in the amount of IL-1β after 4 and 7 days when M0 macrophages are directly or indirectly contacted with fibrin glue containing various immunomodulatory drugs according to one embodiment;

[0037] Figures 19A-19B is the fold change of dead cells in SC islets for various immunomodulatory compounds according to one embodiment ( Figure 19A ) and percentage of dead cells ( Figure 19B )

[0038] Figure 19C is a graph showing the percent viability of SC-islets after exposure to various concentrations of MD2-IN-1 or GDC-2394, according to one embodiment; and

[0039] Figures 20A-20B is a graph of the tannic acid from the sintered membrane ( Figure 19A ) and unsintered films ( Figure 19B ) Plot of cumulative release. DETAILED DESCRIPTION

[0040] In the drive of delivering biological products to treat metabolic disorders such as diabetes, different types of implantable therapeutic devices have been engineered. However, in some cases, the implantation of therapeutic devices may cause undesirable foreign body response. Macrophages play a key role in foreign body response (FBR). Although classical activated macrophages (M1) can be relevant to generating proinflammatory responses (for example, by generating proinflammatory cytokines such as IL-1 β), alternative activated macrophages (M2) can be relevant to generating anti-inflammatory responses (for example, by generating anti-inflammatory cytokines such as IL-10). During use, therapeutic devices may usually trigger undesirable proinflammatory responses (for example, by excessively producing proinflammatory cytokines), which can drive and form undesirable collagen layers around the device, which may subsequently cause the encapsulated cells in the device to suffer from hypoxia. In addition, during use, it may be difficult to regulate the foreign body responses (for example, proinflammatory responses and anti-inflammatory responses) of implantable therapeutic devices.

[0041] In view of the above, the inventors have recognized that while promoting angiogenesis and vascularization, the benefits associated with weakening potential foreign body responses are associated with the use of large encapsulated devices. For example, the large encapsulated device of the present disclosure may contain at least one immunomodulatory compound or be otherwise associated with the at least one immunomodulatory compound, which is capable of inhibiting or reducing the proinflammatory response associated with exposing the device to tissue. In some embodiments, the at least one immunomodulatory compound can be provided in the form of an immunomodulatory material, as described in further detail below. In some cases, the immunomodulatory compound can have a particularly advantageous composition with low cytotoxicity and / or can promote cell viability. The at least one immunomodulatory compound can be capable of inhibiting the proinflammatory response of macrophages (e.g., M0 and / or M1 macrophages). For example, the at least one immunomodulatory compound can be selected from the following groups: toll-like receptor 4 (TLR4) inhibitors, nuclear factor kappa light chain enhancer of activated B cells (NF-κB) inhibitors, colony stimulating factor 1 receptor (CSF1R) inhibitors, and NOD, LRR, and pyrin domain-containing protein 3 (NLRP3) inflammasome inhibitors. These immunomodulatory compounds can be used alone and / or two or more of the immunomodulatory compounds can be used in combination with one another. As described in more detail below, the immunomodulatory compounds can be associated with the macroencapsulated device and portions thereof in any of a variety of suitable ways, including, for example: applied to a tissue adjacent to which the macroencapsulated device is positioned; disposed within the macroencapsulated device; disposed in or on one or more membrane layers of the macroencapsulated device; and / or in any other manner.

[0042] In some embodiments, immunomodulatory compounds and materials described herein can be associated with large encapsulated devices. For example, in one group of embodiments, a large encapsulated device can include multiple membrane layers, such as a first membrane layer and a second membrane layer placed on the first membrane layer. According to some embodiments, the first membrane layer and the second membrane layer can be bonded together to form a seal that extends at least partially around the internal volume placed between the first membrane layer and the second membrane layer. In some cases, the first membrane layer and the second membrane layer can be bonded together to form a seal that extends completely around the internal volume placed between the first membrane layer and the second membrane layer. The seal can be formed in any appropriate position on the membrane layer, such as along the periphery of the membrane layer. In some embodiments, the internal volume can be configured to accommodate a cell colony. The first membrane layer and the second membrane layer can be configured to prevent a cell colony from passing through the device. Any cell of the various appropriate types of cells described elsewhere herein can be placed in the internal volume of the device.

[0043] In some embodiments, the membrane layer can be porous in nature. For example, the first membrane layer and / or the second membrane layer can include a plurality of pores. According to some embodiments, the pores can allow the biological product and / or one or more immunomodulatory compounds to be transported through the membrane layer. The membrane layer can have any of a variety of suitable material properties, including specific material composition, membrane thickness, pore size, etc., as described in more detail below.

[0044] In some aspects, a method for delivering a therapeutic composition produced by a cell population is provided. In some cases, a macroencapsulated device described herein containing a cell population within its interior volume can be used to deliver a therapeutic composition produced by the cells.

[0045] In some embodiments, the method further comprises exposing the tissue surrounding the macroencapsulated device to an immunomodulatory material and / or compound as described herein. The immunomodulatory material and / or compound can be exposed to the tissue by any of a variety of suitable methods. For example, in embodiments in which the immunomodulatory material is disposed on the surface and / or within the pores of one or more membrane layers, the immunomodulatory compound described herein can be released from the surface and / or pores of the one or more membrane layers prior to exposure to the tissue surrounding the at least one membrane layer.

[0046] In some embodiments, a method of treating a subject having a disease is provided. In some embodiments, the method of treating the disease comprises administering a composition described herein to a patient in need thereof. In some embodiments, the disease is type 1 diabetes. In some embodiments, the method of treating type 1 diabetes comprises administering a composition comprising SC-β cells described herein and one or more immunomodulatory compounds to a patient in need thereof. In some embodiments, the method of treating type 1 diabetes comprises administering SC-β cells contained in a device described herein and one or more immunomodulatory compounds described herein to a patient in need thereof. In some embodiments, the method of treating type 1 diabetes comprises administering SC-β cells and one or more immunomodulatory compounds to a patient in need thereof in the absence of a device.

[0047] In embodiments where the immunomodulatory material or compound is fixed to one or more membrane layers via one or more linker molecules, the immunomodulatory compound can be released from the one or more membrane layers upon degradation of the linker molecules at the implantation site. According to some embodiments, the one or more linker molecules can undergo some type of stimulus-responsive degradation, such as enzymatic degradation, hydrolysis, temperature and / or pH-induced degradation, etc. The released immunomodulatory material or compound can then be exposed to tissue surrounding the one or more membrane layers.

[0048] In some embodiments, the immunomodulatory material or compound can be applied directly to the tissue at the implantation area surrounding the one or more membrane layers, rather than being released from the one or more membrane layers. In some such embodiments, the immunomodulatory material or compound can be released into the surrounding tissue when applied directly to the implantation site. Upon release, the immunomodulatory material or compound can be exposed to the tissue surrounding the at least one membrane layer and generate a favorable foreign body response.

[0049] It should be understood that exposure of the immunomodulatory material or compound and diffusion of the therapeutic composition to the surrounding tissue at the implant site can occur in any suitable order, depending on parameters such as the position of the immunomodulatory material or compound relative to the one or more membrane layers, the type of matrix used to encapsulate the compound, etc. For example, in one set of embodiments, both the therapeutic composition and the immunomodulatory material or compound can be exposed to or released to the surrounding tissue simultaneously. Alternatively, the therapeutic composition can diffuse into the surrounding tissue before exposing the immunomodulatory material or compound to the tissue, or vice versa.

[0050] As described above, the methods and devices disclosed herein may include any of a variety of suitable immunomodulatory compounds. According to some embodiments, the immunomodulatory compound can be configured to promote the production of one or more anti-inflammatory cytokines (e.g., IL-10) while inhibiting the production of one or more pro-inflammatory cytokines (e.g., IL-1β). Non-limiting examples of suitable immunomodulatory compounds may include at least one selected from the following groups: toll-like receptor 4 (TLR4) inhibitors, nuclear factor kappa light-chain enhancer of activated B cells (NF-κB) inhibitors, colony stimulating factor 1 receptor (CSF1R) inhibitors, NOD, LRR, and pyrin domain-containing protein 3 (NLRP3) inflammasome inhibitors, and combinations thereof. In other embodiments, immunomodulatory compounds may include Janus kinase inhibitors (JAK) inhibitors (e.g., baricitinib, ruxolitinib), phosphodiesterase-4 (PDE4) inhibitors (e.g., roflumilast, apremilast), CC-chemokine receptor-2 (CCR2) inhibitors (e.g., PF-04136309), phenolic compounds (e.g., polyphenols), toll-like receptor antagonists, IL-1β receptor antagonists (e.g., anakinra), apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain (ASC) inhibitors, caspase inhibitors, gasdermin D (GSDMD) inhibitors, potassium and chloride channel inhibitors and / or nuclear factor erythroid 2-related factor 2 (NFR2) agonists. In particular, without wishing to be bound by any particular theory, it is believed that, under otherwise identical conditions, macroencapsulated devices used with and / or containing (a) certain types of immunomodulatory compounds (e.g., CSF1R inhibitors, NLRP3 inflammasome inhibitors, TLR4 inhibitors, and / or NF-κB inhibitors) can exhibit enhanced reduction of foreign body responses compared to other types of immunomodulatory compounds. In some embodiments, macroencapsulated devices can advantageously include one or more immunomodulatory compounds described elsewhere herein, such as CSF1R inhibitors (e.g., pegidatinib), NLRP3 inflammasome inhibitors (e.g., MCC950), TLR4 inhibitors (e.g., TAK242 and / or tannic acid), and / or NF-κB inhibitors (e.g., TAK242 and / or tannic acid).

[0051] The at least one immunomodulatory compound can be present in the macroencapsulated device or portion thereof at any of a variety of suitable concentrations. For example, the at least one immunomodulatory compound can be present in the matrix, the at least one membrane layer, and / or the device at a concentration greater than or equal to 0.005 mmol / L, greater than or equal to 0.01 mmol / L, greater than or equal to 0.02 mmol / L, greater than or equal to 0.04 mmol / L, greater than or equal to 0.1 mmol / L, greater than or equal to 0.2 mmol / L, greater than or equal to 0.3 mmol / L, greater than or equal to 0.8 mmol / L, greater than or equal to 2 mmol / L, greater than or equal to 4 mmol / L, greater than or equal to 6 mmol / L, greater than or equal to 8 mmol / L, greater than or equal to 10 mmol / L, or greater than or equal to 12 mmol / L. In some embodiments, the at least one immunomodulatory compound is present in the matrix, the at least one membrane layer, and / or the device at a concentration of less than or equal to 20 mmol / L, less than or equal to 15 mmol / L, less than or equal to 12 mmol / L, less than or equal to 10 mmol / L, less than or equal to 8 mmol / L, less than or equal to 6 mmol / L, less than or equal to 4 mmol / L, less than or equal to 2 mmol / L, less than or equal to 1 mmol / L, less than or equal to 0.8 mmol / L, less than or equal to 0.3 mmol / L, less than or equal to 0.1 mmol / L, less than or equal to 0.04 mmol / L, less than or equal to 0.02 mmol / L, or less than or equal to 0.01 mmol / L. Combinations of the above-referenced ranges are possible (e.g., greater than or equal to 0.005 mg and less than or equal to 12 mmol / L).For example, in some embodiments, the at least one immunomodulatory compound may be present in the matrix, the at least one membrane layer, and / or the device at a concentration of 0.005-15 mmol / L, 0.005-12 mmol / L, 0.005-6 mmol / L, 0.005-4 mmol / L, 0.005-2 mmol / L, 0.005-1 mmol / L, 0.005-0.8 mmol / L, 0.01-15 mmol / L, 0.01-12 mmol / L, 0.01-6 mmol / L, 0.01-4 mmol / L, 0.01-2 mmol / L, 0.01-1 mmol / L, 0.01-0.8 mmol / L, 0.02-15 mmol / L, 0.02-12 mmol / L, 0.02-6 mmol / L, 0.02-4 mmol / L l / L, 0.02-2mmol / L, 0.02-1mmol / L, 0.02-0.8mmol / L, 0.04-12mmol / L, 0.04-6mmol / L, 0.0 4-4mmol / L, 0.04-2mmol / L, 0.04-1mmol / L, 0.04-0.8mmol / L, 0.1-15mmol / L, 0.1-12mmol / L , 0.1-6mmol / L, 0.1-4mmol / L, 0.1-2mmol / L, 0.1-1mmol / L, 0.1-0.8mmol / L, 0.3-15mmol / L , 0.3-12mmol / L, 0.3-6mmol / L, 0.3-4mmol / L, 0.3-2mmol / L, 0.3-1mmol / L or 0.3-0.8mmol / L. In some embodiments, the at least one immunomodulatory compound may be present in the matrix, the at least one film layer and / or the device at a concentration greater than or equal to 0.01 mmol / L and less than or equal to 0.8 mmol / L. In some embodiments, the at least one immunomodulatory compound may be present in the matrix, the at least one film layer and / or the device at a concentration greater than or equal to 0.04 mmol / L and less than or equal to 4 mmol / L. In some embodiments, the at least one immunomodulatory compound may be present in the matrix, the at least one film layer and / or the device at a concentration greater than or equal to 0.02 mmol / L and less than or equal to 2 mmol / L. In some embodiments, the at least one immunomodulatory compound may be present in the matrix, the at least one film layer and / or the device at a concentration greater than or equal to 0.3 mmol / L and less than or equal to 12 mmol / L. Other ranges are also possible.

[0052] The at least one immunomodulatory compound can be used in any therapeutic dose of various appropriate therapeutic doses. For example, in some embodiments, the therapeutic dose can be measured as the amount (for example, in micromoles) of the immunomodulatory compound per volume (for example, per liter of tissue). In some embodiments, the immunomodulatory compound can be used in the following therapeutic doses: greater than or equal to 0.05 μM, greater than or equal to 0.1 μM, greater than or equal to 0.125 μM, greater than or equal to 0.15 μM, greater than or equal to 0.25 μM, greater than or equal to 0.5 μM, greater than or equal to 1 μM, greater than or equal to 2 μM, greater than or equal to 4 μM or more and / or less than or equal to 10 μM, less than or equal to 6 μM, less than or equal to 4 μM, less than or equal to 2 μM, less than or equal to 1 μM, less than or equal to 0.5 μM, less than or equal to 0.25 μM, less than or equal to 0.15 μM, less than or equal to 0.125 μM, less than or equal to 0.1 μM or less. Combinations of the above-referenced ranges are possible (e.g., greater than or equal to 0.1 μM and less than or equal to 4 μM). For example, in some embodiments, the immunomodulatory compound can be used in the following therapeutic doses: 0.1-6 μM, 0.1-4 μM, 0.1-3.5 μM, 0.1-3 μM, 0.1-2.5 μM, 0.1-2 μM, 0.1-1.5 μM, 0.1-1 μM, 0.1-0.5 μM, 0.1-0.25 μM, 0.25-6 μM, 0.25-4 μM, 0.25-3.5 μM, 0.25-3 μM, 0.25-2.5 μM, 0.25-2 μM, 0. In some embodiments, the immunomodulatory compound can be used at a therapeutic dose of greater than or equal to 0.125 μM and less than or equal to 0.25 μM. In some embodiments, the immunomodulatory compound can be used at a therapeutic dose of greater than or equal to 0.25 μM and less than or equal to 4 μM. In some embodiments, the immunomodulatory compound can be used at a therapeutic dose greater than or equal to 2 μM and less than or equal to 4 μM. In some embodiments, the immunomodulatory compound can be used at a therapeutic dose greater than or equal to 0.25 μM and less than or equal to 0.50 μM. Other ranges are also possible.

[0053] In one set of embodiments, the immunomodulatory compound comprises a colony stimulating factor 1 receptor (CSF1R) inhibitor. Any of a variety of CSF1R inhibitors can be used, including but not limited to pecidinib or a salt, analog, or derivative thereof. In one set of embodiments, the immunomodulatory compound comprises pecidinib. According to some embodiments, under otherwise identical conditions, the presence of a particular type of CSF1R inhibitor (e.g., pecidinib) in a macroencapsulated device can result in a relatively high reduction in foreign body response compared to other types of CSF1R inhibitors. In some embodiments, the one or more CSF1R inhibitors can be present in the matrix at a concentration between or equal to 0.01 mmol / L and less than or equal to 0.8 mmol / L. In some embodiments, the one or more CSF1R inhibitors can be present in the matrix at a concentration between or equal to 0.005 mmol / L and less than or equal to 1 mmol / L. In some embodiments, the one or more CSF1R inhibitors can be present in the matrix at a concentration between or equal to 0.005 mmol / L and less than or equal to 1.5 mmol / L. In addition, one or more CSF1R inhibitors can be used at a therapeutic dose between or equal to 0.25 μM and less than or equal to 4 μM. In some embodiments, one or more CSF1R inhibitors can be used at a therapeutic dose between or equal to 0.125 μM and less than or equal to 0.25 μM. In some embodiments, one or more CSF1R inhibitors can be used at a therapeutic dose between or equal to 0.1 μM and less than or equal to 0.35 μM. In some embodiments, one or more CSF1R inhibitors can be used at a therapeutic dose between or equal to 0.05 μM and less than or equal to 0.5 μM. Of course, it is also contemplated to use CSF1R inhibitors at concentrations and / or therapeutic doses greater than or less than those described above, including the ranges of general immunomodulatory compounds described above, as the present disclosure is not limited thereto.

[0054] Alternatively or additionally, according to some embodiments, the immunomodulatory compound comprises an NLRP3 inflammasome inhibitor. Any NLRP3 inhibitor among various NLRP3 inhibitors can be used, including but not limited to MCC950, GDC-2394, YQ128, NLRP3 inflammasome inhibitor I, musk ketone, 4'-methoxyresveratrol, CY-09, INF39, dapansutrile, sterol, licorice chalcone B and dimethyl itaconate or its salt, analog or derivative. In one group of embodiments, the immunomodulatory compound comprises MCC950. In some embodiments, the immunomodulatory compound comprises GDC-2394. In some embodiments, the immunomodulatory compound comprises YQ128. In some embodiments, the immunomodulatory compound comprises NLRP3 inflammasome inhibitor I. In some embodiments, the immunomodulatory compound comprises musk ketone, 4'-methoxyresveratrol. In some embodiments, the immunomodulatory compound comprises CY-09. In some embodiments, the immunomodulatory compound comprises INF39. In some embodiments, the immunomodulatory compound comprises dapansutrile. In some embodiments, the immunomodulatory compound comprises asterone. In some embodiments, the immunomodulatory compound comprises licorice chalcone B. In some embodiments, the immunomodulatory compound comprises dimethyl itaconate. According to some embodiments, under otherwise identical conditions, the presence of a specific type of NLPR3 inhibitor (e.g., MCC950) in a large encapsulated device can cause a relatively high foreign body response reduction compared to other types of NLPR3 inhibitors. In some embodiments, one or more NLRP3 inflammasome inhibitors may be present in the matrix at a concentration between or equal to 0.04 mmol / L and less than or equal to 4 mmol / L. In some embodiments, one or more NLRP3 inflammasome inhibitors may be present in the matrix at a concentration between or equal to 0.02 mmol / L and less than or equal to 6 mmol / L. In some embodiments, one or more NLRP3 inflammasome inhibitors may be present in the matrix at a concentration between or equal to 0.01 mmol / L and less than or equal to 8 mmol / L. In addition, one or more NLRP3 inflammasome inhibitors may be used in a therapeutic dose between or equal to 0.25 μM and less than or equal to 4 μM. In some embodiments, one or more NLRP3 inflammasome inhibitors may be used at a therapeutic dose between 0.2 μM and less than or equal to 5 μM. In some embodiments, one or more NLRP3 inflammasome inhibitors may be used at a therapeutic dose between 0.15 μM and less than or equal to 6 μM. Of course, it is also contemplated to use NLRP3 inflammasome inhibitors at concentrations and / or therapeutic doses greater than or less than those described above, including the range of general immunomodulatory compounds described above, as the present disclosure is not limited thereto.

[0055] Alternatively or additionally, in some embodiments, the immunomodulatory compound comprises an NLRP3 inhibitor comprising GDC-2394. In some embodiments, the immunomodulatory compound comprises an NLRP3 inhibitor comprising YQ128. In some embodiments, the immunomodulatory compound comprises an NLRP3 inhibitor comprising NLRP3 inflammasome inhibitor I. In some embodiments, the immunomodulatory compound comprises an NLRP3 inhibitor comprising musk ketone. In some embodiments, the immunomodulatory compound comprises an NLRP3 inhibitor comprising 4'-methoxyresveratrol. In some embodiments, the immunomodulatory compound comprises an NLRP3 inhibitor comprising CY-09. In some embodiments, the immunomodulatory compound comprises an NLRP3 inhibitor comprising INF39. In some embodiments, the immunomodulatory compound comprises an NLRP3 inhibitor comprising dapamsulfuronil. In some embodiments, the immunomodulatory compound comprises an NLRP3 inhibitor comprising sterone. In some embodiments, the immunomodulatory compound comprises an NLRP3 inhibitor comprising licochalcone B. In some embodiments, the immunomodulatory compound comprises an NLRP3 inhibitor comprising dimethyl itaconate.

[0056] Alternatively or in addition, according to some embodiments, the immunomodulatory compound comprises a toll-like receptor 4 (TLR4) inhibitor. Any of a variety of TLR4 inhibitors can be used, including but not limited to TAK242, tannic acid, MD2-IN-1, TLR4-IN-C34, IAXO-102 and / or MD2-TLR4-IN-1 or a salt, analog or derivative thereof. In one group of embodiments, the immunomodulatory compound comprises TAK242. In some embodiments, the immunomodulatory compound comprises tannic acid. In some embodiments, the immunomodulatory compound comprises MD2-IN-1. In some embodiments, the immunomodulatory compound comprises TLR4-IN-C34. In some embodiments, the immunomodulatory compound comprises IAXO-102. In some embodiments, the immunomodulatory compound comprises MD2-TLR4-IN-1. According to some embodiments, under otherwise identical conditions, the presence of a specific type of TLR4 inhibitor (e.g., TAK242 and / or tannic acid) in a macroencapsulated device can cause a relatively high reduction in foreign body response compared to other types of TLR4 inhibitors. In some embodiments, one or more TLR4 inhibitors may be present in the matrix at a concentration between or equal to 0.3 mmol / L and 12 mmol / L. In some embodiments, one or more TLR4 inhibitors may be present in the matrix at a concentration between or equal to 0.2 mmol / L and 15 mmol / L. In some embodiments, one or more TLR4 inhibitors may be present in the matrix at a concentration between or equal to 0.02 mmol / L and 2 mmol / L. In some embodiments, one or more TLR4 inhibitors may be present in the matrix at a concentration between or equal to 0.01 mmol / L and 4 mmol / L. In addition, one or more TLR4 inhibitors may be used in a therapeutic dose between or equal to 2 μM and less than or equal to 4 μM. In some embodiments, one or more TLR4 inhibitors may be used in a therapeutic dose between or equal to 1 μM and less than or equal to 6 μM. In some embodiments, one or more TLR4 inhibitors may be used in a therapeutic dose between or equal to 0.25 μM and less than or equal to 0.5 μM. In some embodiments, one or more TLR4 inhibitors can be used in a therapeutic dose between 0.15 μM and less than or equal to 1 μM. Of course, it is also contemplated to use TLR4 inhibitors at concentrations and / or therapeutic doses greater than or less than those described above, including the ranges of general immunomodulatory compounds described above, as the present disclosure is not limited thereto.

[0057] Alternatively or additionally, in some embodiments, the immunomodulatory compound comprises a TLR4 inhibitor comprising TLR4-IN-C34. In some embodiments, the immunomodulatory compound comprises a TLR4 inhibitor comprising IAXO-102. In some embodiments, the immunomodulatory compound comprises a TLR4 inhibitor comprising MD2-TLR4-IN-1.

[0058] Alternatively or in addition, according to some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor. Any of the various NF-κB inhibitors can be used, including but not limited to TAK242, tannic acid, sitagliptin, cornus glycosides, SM-7368, NF-κB-IN-1, IQ 3, sulfasalazine (NSC 667219), erdosteine, evodiamine, UCB-9260, triptolide (PG490), curcuminol, SN50, INH14, SC75741, JSH-23, luteolin, olive acid, morulin A, eleutheroside E and / or madecassic acid or its salt, analog or derivative. In one group of embodiments, the immunomodulatory compound comprises an NF-κB inhibitor including TAK242. In one group of embodiments, the immunomodulatory compound comprises an NF-κB inhibitor including tannic acid. In one set of embodiments, the immunomodulatory compound comprises TAK242 and / or tannic acid. According to some embodiments, under otherwise identical conditions, the presence of a specific type of NF-κB inhibitor (e.g., TAK242 and / or tannic acid) in a macroencapsulated device can result in a relatively high reduction in foreign body response compared to other types of NF-κB inhibitors. In some embodiments, one or more NF-κB inhibitors can be present in the matrix at a concentration between or equal to 0.3 mmol / L and 12 mmol / L. In some embodiments, one or more NF-κB inhibitors can be present in the matrix at a concentration between or equal to 0.2 mmol / L and 15 mmol / L. In some embodiments, one or more NF-κB inhibitors can be present in the matrix at a concentration between or equal to 0.02 mmol / L and 2 mmol / L. In some embodiments, one or more NF-κB inhibitors can be present in the matrix at a concentration between or equal to 0.01 mmol / L and 4 mmol / L. In addition, one or more NF-κB inhibitors can be used in a therapeutic dose between or equal to 2 μM and less than or equal to 4 μM. In some embodiments, one or more NF-κB inhibitors may be used at a therapeutic dose between 1 μM and less than or equal to 6 μM. In some embodiments, one or more NF-κB inhibitors may be used at a therapeutic dose between 0.25 μM and less than or equal to 0.5 μM. In some embodiments, one or more NF-κB inhibitors may be used at a therapeutic dose between 0.15 μM and less than or equal to 1 μM. Of course, it is also contemplated to use NF-κB inhibitors at concentrations and / or therapeutic doses greater than or less than those described above, including the range of general immunomodulatory compounds described above, as the present disclosure is not limited thereto.

[0059] Alternatively or additionally, in some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising sitagliptin. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising cornusin. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising SM-7368. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising NF-κB-IN-1. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising IQ 3. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising sulfasalazine (NSC 667219). In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising erdosteine. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising evodiamine. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising UCB-9260. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising triptolide (PG490). In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising curcuminol. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising SN50. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising INH14. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising SC75741. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising JSH-23. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising luteolin. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising oliveric acid. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising moruproside A. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising eleutheroside E. In some embodiments, the immunomodulatory compound comprises an NF-κB inhibitor comprising madecassic acid.

[0060] Alternatively or in addition, according to some embodiments, the immunomodulatory compound comprises a TLR-4 and / or NF-κB inhibitor (e.g., TAK242 and / or tannic acid). In some embodiments, the immunomodulatory compound comprises a TLR-4 and NF-κB inhibitor, such as TAK242. In some embodiments, the immunomodulatory compound comprises a TLR-4 and NF-κB inhibitor, such as tannic acid. According to some embodiments, the TLR-4 and / or NF-κB inhibitor comprises a polyphenol with anti-inflammatory and / or redox scavenging properties. In one set of embodiments, the immunomodulatory compound comprises tannic acid. In some embodiments, the tannic acid may be present in the matrix at a concentration between or equal to 0.02 mmol / L and 2 mmol / L. In addition, tannic acid may be used in a therapeutic dose between or equal to 0.25 μM and less than or equal to 0.5 μM. In one set of embodiments, the immunomodulatory compound comprises TAK242. In some embodiments, TAK242 may be present in the matrix at a concentration between or equal to 0.3 mmol / L and 12 mmol / L. Additionally, TAK242 can be used at a therapeutic dose between 2 μM or less and 4 μM or less. Of course, it is also contemplated to use tannic acid and / or TAK242 at concentrations and / or therapeutic doses greater than or less than those described above, including the range of general immunomodulatory compounds described above, as the present disclosure is not limited thereto.

[0061] In some embodiments, the immunomodulatory compound comprises a Janus kinase inhibitor (JAK) inhibitor (e.g., baricitinib, ruxolitinib). In some embodiments, the immunomodulatory compound comprises a phosphodiesterase-4 (PDE4) inhibitor (e.g., roflumilast, apremilast). In some embodiments, the immunomodulatory compound comprises a CC-chemokine receptor-2 (CCR2) inhibitor (e.g., PF-04136309). In some embodiments, the immunomodulatory compound comprises a phenolic compound (e.g., a polyphenol). In some embodiments, the immunomodulatory compound comprises a toll-like receptor antagonist. In some embodiments, the immunomodulatory compound comprises an IL-1β receptor antagonist (e.g., anakinra). In some embodiments, the immunomodulatory compound comprises an apoptosis-associated speck-like protein (ASC) inhibitor containing a C-terminal caspase recruitment domain. In some embodiments, the immunomodulatory compound comprises a caspase inhibitor. In some embodiments, the immunomodulatory compound comprises a gasdermin D (GSDMD) inhibitor. In some embodiments, the immunomodulatory compound comprises a potassium and chloride channel inhibitor. In some embodiments, the immunomodulatory compound comprises a nuclear factor erythroid 2-related factor 2 (NFR2) agonist.

[0062] In some embodiments, the immunomodulatory material can comprise a matrix in which the at least one immunomodulatory compound is dispersed or otherwise included. According to some embodiments, the matrix can comprise a hydrogel, microparticles, nanoparticles, a biodegradable material, and / or a swellable polymer. The at least one immunomodulatory compound can be associated with the matrix in any of a variety of suitable ways, such as being contained within and / or conjugated to the matrix. The matrix can comprise any of the various suitable compositions described elsewhere herein.

[0063] The immunomodulatory material may be present in any of the various suitable locations in the large encapsulated device. According to some embodiments, the immunomodulatory material may be associated with one or more membrane layers (e.g., the first membrane layer and / or the second membrane layer) of the large encapsulated device in any of the various suitable ways described herein. In one group of embodiments, the immunomodulatory material is placed on one or more membrane layers in the membrane layer, such as on the first membrane layer and / or the second membrane layer. For example, according to one embodiment, the immunomodulatory material may be placed on the surface of one or more membrane layers as a surface coating. The immunomodulatory material may be used as a matrix containing an immunomodulatory compound or as an immunomodulatory compound itself (without a matrix) surface coating on one or more membrane layers. In some cases, the surface coating may be applied to the inner surface and / or outer surface of one or more membrane layers. Without wishing to be bound by any particular theory, it is believed that applying the immunomodulatory material to the outer surface of one or more membrane layers may be advantageous. This may allow the immunomodulatory material to be in direct contact with surrounding tissues during implantation and contribute to a more favorable foreign body response.

[0064] Alternatively or in addition, immunomodulatory material can be placed on one or more film layers by chemical fixation to the surface of one or more film layers. For example, in one embodiment, immunomodulatory compounds described herein can be fixed to the surface (for example, outer surface) of one or more film layers by chemical conjugation. In some cases, one or more linker molecules can be used to fix immunomodulatory compounds to one or more film layers. In certain embodiments, one or more linker molecules can be biodegradable and can be easily degraded, such as enzymatic degradation, hydrolysis, pH and / or temperature-induced degradation, etc. Immunomodulatory compounds can be directly conjugated to one or more film layers and / or included in a matrix that is then conjugated to one or more film layers. Alternatively or in addition, immunomodulatory material can be placed in the pores of at least one film layer in (for example, impregnation) one or more film layers, such as in the pores of the first film layer and / or the second film layer. Alternatively or in addition, immunomodulatory material can be placed in the internal volume of the device formed between one or more film layers.

[0065] Alternatively or in addition, the immunomodulatory material can be placed adjacent to one or more membrane layers, such as adjacent to the first membrane layer and / or the second membrane layer. In one embodiment, the immunomodulatory material can be placed around at least a portion of one or more membrane layers. For example, the immunomodulatory material can be placed around at least a portion of the periphery of one or more layers. In some cases, the immunomodulatory material can be applied to tissue within the surgical area (during implantation), which can be located near a large encapsulated device and may be placed against the large encapsulated device after implantation. According to some embodiments, the immunomodulatory material is applied to the membrane and / or adjacent positions using the methods described above, for example, on the outer surface of the membrane layer and / or directly applied to the tissue area around the large encapsulated device, so that the immunomodulatory compound can be in direct contact with the tissue during implantation and may be advantageous. Such application methods may cause the favorable foreign body response described elsewhere herein.

[0066] As described above, in some embodiments, the immunomodulatory material comprises a matrix in which the immunomodulatory compound can be dispersed. In some cases, the matrix comprises a hydrogel. According to some embodiments, the term hydrogel can refer to a gel in which a cross-linked polymer matrix is ​​completely or partially swollen by water, one or more water-compatible alcohols or a combination thereof. The polymer matrix can be cross-linked by chemical or physical means. As a non-limiting example, the hydrogel can be cross-linked by covalent bonds, ionic interactions, hydrogen bonding, chain entanglement or self-association of microphase separation parts. In addition, it should be understood that such hydrogels can exist and be used in a dehydrated (e.g., unswollen or lyophilized) state.

[0067] In some embodiments, the hydrogel may comprise a polymer matrix selected from at least one water-compatible organic polymer, an alcohol-compatible organic polymer, and combinations thereof. The polymer may be homopolymeric, heteropolymeric (including but not limited to any copolymer distribution of cross-linked polymers or copolymers), and may be linear, branched, hyperbranched, dendritic, or cross-linked to any extent. Examples of suitable polymers include, but are not limited to, gelatin, methylcellulose, hydroxyethyl methylcellulose, hydroxypropyl methylcellulose, polyethylene glycol, polyethylene oxide, polyacrylamide, polyacrylic acid, polymethacrylic acid, salts of polyacrylic acid, salts of polymethacrylic acid, poly(2-hydroxyethyl methacrylate), polycaprolactone, polylactic acid, polyglycolic acid, polylactic-co-glycolic acid, polyvinyl alcohol, polyanhydrides such as poly(methacrylic) anhydride, poly(acrylic) anhydride, polysebacic anhydride, collagen, keratin, poly(hyaluronic acid), hyaluronic acid-containing polymers and copolymers, polypeptides, dextran, dextran sulfate, chitosan, chitin, agarose gel, fibrin gel, soy-derived hydrogels, and alginate-based hydrogels such as poly(sodium alginate), and combinations thereof.

[0068] In some embodiments, the hydrogel polymer matrix can include any of a variety of biodegradable and / or biocompatible polymers, such as biopolymers (or natural polymers) and / or biodegradable and / or biocompatible synthetic polymers. Examples of suitable biodegradable polymers include synthetic polymers such as polymers of lactic acid and glycolic acid, polyanhydrides, poly(ortho)esters, polyurethanes, poly(butyric acid), poly(valeric acid), poly(caprolactone), poly(hydroxybutyrate), poly(lactide-co-glycolide) and poly(lactide-co-caprolactone), polycaprolactone; and natural polymers such as polysaccharides (e.g., starch, alginate, dextran, agarose, cellulose, etc.), chemical derivatives thereof (substitution, addition of chemical groups such as alkyl, alkylene, hydroxylation, oxidation, and other modifications routinely performed by those skilled in the art), proteins such as gelatin, collagen, keratin, fibrinogen, albumin, albumin and other hydrophilic proteins, zein and other alcohol-soluble proteins and hydrophobic proteins, copolymers, and mixtures thereof. According to some embodiments, biodegradable polymers can be degraded by enzymatic hydrolysis or exposure to water, by surface or body erosion. The hydrogel matrix can contain a polymer or more than a polymer. When there is more than a polymer, the continuous matrix can contain a mixture of a blend of polymers or a copolymer containing a polymer. In certain embodiments, the biodegradable matrix material can be used to release the relevant immunomodulatory compounds when the matrix material is degraded in situ by any degradation pathway in a variety of degradation pathways, such as enzymatic degradation, hydrolysis, pH and / or temperature-induced degradation.

[0069] Specific examples of materials (e.g., hydrogels and / or polymers) that can be mixed with an immunomodulatory compound and / or used to encapsulate the immunomodulatory compound to provide the desired controlled release of the compound can include, but are not limited to, fibrin polymers, alginate, collagen gel, poly(2-hydroxyethyl methacrylate) (pHEMA), poly(2-hydroxypropyl acrylate) (pHPA), poly(lactic-co-glycolic acid) (PLGA), agarose, chitosan, carboxymethyl cellulose, poly(vinyl alcohol) (PVA), Eudragit, poly(methyl methacrylate) (PMMA), polylactic acid (PLA), polycaprolactone (PCL), polyacrylonitrile (PAN), PAN / PVC blends or copolymers, polyurethane, collagen, laminin, polyethylene glycol (PEG), hydroxyapatite, hydroxypropyl cellulose, Pluronics (e.g., F127), microsphere slurries, carbon nanotubes, and / or any other suitable material described elsewhere herein.

[0070] The matrix can be present in any of the various suitable shapes and / or forms described elsewhere herein. In one set of embodiments, the matrix can be in the form of particles, such as microparticles and / or nanoparticles. For example, the matrix can be in the form of hydrogel particles. In addition, it should be understood that the hydrogel particles can include both polymer particles swollen with a fully compatible fluid and polymer particles in a dehydrated state. In one set of embodiments, the polymer particles are PEG and / or PLGA particles. In some cases, one or more immunomodulatory compounds described herein can be contained or encapsulated by the particles, or in some cases, can be chemically conjugated to the particles.

[0071] In some embodiments, the hydrogel is an injectable and / or flowable gel. That is, the injectable and / or flowable gel can be deformed and undergo flow when a force (e.g., shear force) is applied. Such gels can be advantageously used with syringes for delivery and local application. For example, in one group of embodiments, the injectable gel comprises a protein gel, such as a fibrin gel.

[0072] Any method in the various methods can be used to form immunomodulatory materials and / or matrix described herein.For example, in one group of embodiments, protein gel, such as the fibrin gel formed by fibrinogen and thrombin, can be used as matrix.In certain embodiments, the fibrin gel precursor mixture can include a liquid carrier (for example, a diluent).For example, fibrinogen can be diluted in a liquid carrier (for example, PBS) before mixing with thrombin.In one embodiment, before mixing with thrombin, fibrinogen can account for at least 50% (for example, at least 60%, at least 70%, at least 80%) and / or at most 90% (for example, at most 95%, at most 99% or at most 100%) of the total volume of the mixture containing fibrinogen and liquid carrier.In one embodiment, fibrinogen can account for at least 50% (for example, at least 60%, at least 70%, at least 80%) and / or at most 90% (for example, at most 95%, at most 99% or at most 100%) of the total volume of the mixture containing fibrinogen and thrombin. In one embodiment, fibrinogen can account for 50-99%, 50-95%, 60-99%, 60-95%, 70-99%, 70-95%, 80-99% or 80-95% of the total volume of the mixture containing fibrinogen and thrombin. One or more immunomodulatory compounds described herein can be present in the matrix mixture with the amount or concentration described elsewhere herein. The mixture can gel at any temperature in various temperatures, such as forming a gel at about 37 ℃.

[0073] In some embodiments, compositions are described herein that can be used without a large encapsulated device, wherein the compositions are directly implanted in the target tissue. According to certain embodiments, the compositions include a cell colony and at least one immunomodulatory compound. The cell colony may include any cell colony in the various appropriate cell colonies described elsewhere herein, alone or in combination with each other, including, for example, NKX6.1 positive and / or ISL1 positive cells. Similarly, immunomodulatory compounds may include any one or more immunomodulatory compounds in the various appropriate immunomodulatory compounds described elsewhere herein, such as toll-like receptor 4 (TLR4) inhibitors (e.g., TAK242 and / or tannic acid), nuclear factor kappa light chain enhancer (NF-κB) inhibitors of activated B cells (e.g., TAK242 and / or tannic acid), colony stimulating factor 1 receptor (CSF1R) inhibitors (e.g., Pecidinib) and / or protein 3 (NLRP3) inflammasome inhibitors containing NOD, LRR and pyrin domains (e.g., MCC950).

[0074] According to some embodiments, compositions may further include a matrix described elsewhere herein, such as a hydrogel. In certain embodiments, the cell colony may be contained in a matrix, such as encapsulated or dispersed in a matrix. In certain embodiments, the immunomodulatory compound may be associated with the matrix in any of the various appropriate ways described elsewhere herein. For example, in some embodiments, the immunomodulatory compound may be contained in a matrix (e.g., dispersed in, encapsulated in, and / or impregnated in the pores of the matrix). Alternatively or in addition, the immunomodulatory compound may be attached to a part of the matrix, for example, by chemical conjugation or physical association. The matrix may include any of the various suitable compositions described elsewhere herein.

[0075] In some embodiments, the apparatus and methods described herein can be used to realize the sustained exposure or release of immunomodulatory materials or compounds. For example, immunomodulatory materials or compounds can be exposed to or released to the tissue at the implantation site over an extended period of time, such as at least 1 day, at least 2 days, at least 4 days, at least 7 days, at least 10 days, at least 14 days or longer and / or at most 21 days, at most 28 days or longer. For example, immunomodulatory materials or compounds can be exposed to or released to the tissue at the implantation site over a period of at least 1 day. In some embodiments, immunomodulatory materials or compounds can be exposed to or released to the tissue at the implantation site over a period of at least 2 days. In some embodiments, immunomodulatory materials or compounds can be exposed to or released to the tissue at the implantation site over a period of at least 4 days. In some embodiments, immunomodulatory materials or compounds can be exposed to or released to the tissue at the implantation site over a period of at least 7 days. In some embodiments, immunomodulatory materials or compounds can be exposed to or released to the tissue at the implantation site over a period of at least 10 days. In some embodiments, immunomodulatory materials or compounds can be exposed to or released to the tissue at the implantation site over a period of at least 14 days or longer. In certain embodiments, immunomodulatory material or compound can be exposed to or released to the tissue at the implantation site within a time period of at most 21 days. In certain embodiments, immunomodulatory material or compound can be exposed to or released to the tissue at the implantation site within a time period of at most 28 days. The combination of the above ranges is possible (e.g., at least 1 day and at most 28 days, or at least 1 day and at most 2 weeks). Other ranges are also possible.

[0076] The present inventors have found that by using immunomodulatory materials and / or macroencapsulated devices as disclosed herein, foreign body responses can be advantageously modulated during the delivery and / or implantation of the macroencapsulated device into the desired location in the subject. For example, by employing the immunomodulatory materials described herein, macrophages can be polarized toward M2 macrophages (relative to M1 macrophages), such that an unfavorable foreign body response can be at least partially attenuated, and angiogenesis and vascularization can be enhanced. The immunomodulatory materials and / or macroencapsulated devices described herein can be associated with many additional advantages, such as sustained release of immunomodulatory compounds over an extended period of time, little or no cytotoxicity, little or almost no disruption to the formation of vascular networks, enhanced angiogenesis and vascularization, enhanced chemical stability associated with immunomodulatory compounds, effective delivery of therapeutic compositions produced by the cell population contained within the macroencapsulated device, and an overall reduction in foreign body responses compared to devices not containing the immunomodulatory materials described herein.

[0077] As described above, using a large encapsulated device comprising an immunomodulatory compound or material as described herein can be associated with an adverse foreign body response, such as an enhanced reduction in proinflammatory response. For example, compared with a conventional large encapsulated device without an immunomodulatory material or compound, the large encapsulated device comprising one or more of the immunomodulatory material or compound as described herein can cause a reduction in the foreign body response during implantation. In some cases, compared with a large encapsulated device without an immunomodulatory material or compound under similar conditions, the large encapsulated device described herein can cause a reduction in proinflammatory response of at least 1.1, 1.3, 1.5, 2, 4, 6, 10, 15, 20 times or other appropriate multiples. In certain embodiments, compared with a large encapsulated device without an immunomodulatory material or compound under similar conditions, the large encapsulated device described herein can show a reduction in proinflammatory response of less than or equal to or at most 100, 50, 40, 30, 20, 15, 10 times or other appropriate multiples. For example, in some embodiments, compared with a large encapsulated device without an immunomodulatory material or compound under similar conditions, the large encapsulated device described herein can show a reduction in proinflammatory response of at most 30 times. In some embodiments, the macroencapsulated devices described herein can exhibit up to a 50-fold reduction in proinflammatory response compared to macroencapsulated devices without immunomodulatory materials or compounds under similar conditions. Combinations of the above ranges are possible (e.g., reductions by a factor between or equal to 1.1 and 11, 1.1 and 25, 1.1 and 30, 1.1 and 50, or other suitable factors). Other ranges are also possible.

[0078] According to some embodiments, the feature of foreign body response can be that when large encapsulated device is exposed to macrophage, the total amount of anti-inflammatory cytokines (e.g., IL-10) generated by macrophage is relative to the total amount of pro-inflammatory cytokines (e.g., IL-1 β). Macrophages can include monocytes, M0 macrophages and / or M1 macrophages. In some embodiments, under otherwise identical conditions, the total amount of anti-inflammatory cytokines (e.g., IL-10) generated by each macrophage in the presence of a large encapsulated device comprising immunomodulatory compounds and / or materials and the total amount of pro-inflammatory cytokines (e.g., IL-1 β) is at least 1.05 times, 1.1 times, at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 8 times, at least 10 times, at least 15 times or at least 20 times the total amount of anti-inflammatory cytokines generated by each macrophage in the presence of a large encapsulated device without immunomodulatory compounds and / or materials and the total amount of pro-inflammatory cytokines. In some embodiments, under otherwise identical conditions, the ratio of the total amount of anti-inflammatory cytokines (e.g., IL-10) to the total amount of pro-inflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device comprising an immunomodulatory compound and / or material described herein is at least 1.1 times greater than the ratio of the total amount of anti-inflammatory cytokines to the total amount of pro-inflammatory cytokines produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or material. In some embodiments, under otherwise identical conditions, the ratio of the total amount of anti-inflammatory cytokines (e.g., IL-10) to the total amount of pro-inflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device comprising an immunomodulatory compound and / or material described herein is at least 1.5 times greater than the ratio of the total amount of anti-inflammatory cytokines to the total amount of pro-inflammatory cytokines produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or material. In some embodiments, under otherwise identical conditions, the ratio of the total amount of anti-inflammatory cytokines (e.g., IL-10) to the total amount of pro-inflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device comprising an immunomodulatory compound and / or material described herein is at least 2-fold the ratio of the total amount of anti-inflammatory cytokines to the total amount of pro-inflammatory cytokines produced per macrophage in the presence of a macroencapsulated device without the immunomodulatory compound and / or material.In some embodiments, under otherwise identical conditions, the ratio of the total amount of anti-inflammatory cytokines (e.g., IL-10) to the total amount of pro-inflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device comprising an immunomodulatory compound and / or material described herein is at least 3 times greater than the ratio of the total amount of anti-inflammatory cytokines to the total amount of pro-inflammatory cytokines produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or material. In some embodiments, under otherwise identical conditions, the ratio of the total amount of anti-inflammatory cytokines (e.g., IL-10) to the total amount of pro-inflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device comprising an immunomodulatory compound and / or material described herein is at least 4 times greater than the ratio of the total amount of anti-inflammatory cytokines to the total amount of pro-inflammatory cytokines produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or material. In some embodiments, under otherwise identical conditions, the ratio of the total amount of anti-inflammatory cytokines (e.g., IL-10) produced per macrophage to the total amount of pro-inflammatory cytokines (e.g., IL-1β) produced in the presence of a macroencapsulated device containing an immunomodulatory compound and / or material described herein is at least 5 times greater than the ratio of the total amount of anti-inflammatory cytokines to the total amount of pro-inflammatory cytokines produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials. In some embodiments, under otherwise identical conditions, the ratio of the total amount of anti-inflammatory cytokines (e.g., IL-10) produced per macrophage to the total amount of pro-inflammatory cytokines (e.g., IL-1β) produced in the presence of a macroencapsulated device containing an immunomodulatory compound and / or material described herein is at most 10 times, at most 20 times, at most 25 times, at most 30 times, at most 50 times, at most 75 times, or at most 100 times greater than the ratio of the total amount of anti-inflammatory cytokines to the total amount of pro-inflammatory cytokines produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials. In some embodiments, under otherwise identical conditions, the ratio of the total amount of anti-inflammatory cytokines (e.g., IL-10) to the total amount of pro-inflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing an immunomodulatory compound and / or material described herein is at most 10 times the ratio of the total amount of anti-inflammatory cytokines to the total amount of pro-inflammatory cytokines produced per macrophage in the presence of a macroencapsulated device without an immunomodulatory compound and / or material.In some embodiments, under otherwise identical conditions, the ratio of the total amount of anti-inflammatory cytokines (e.g., IL-10) to the total amount of pro-inflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein is at most 20 times the ratio of the total amount of anti-inflammatory cytokines to the total amount of pro-inflammatory cytokines produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials. In some embodiments, under otherwise identical conditions, the ratio of the total amount of anti-inflammatory cytokines (e.g., IL-10) to the total amount of pro-inflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein is at most 25 times the ratio of the total amount of anti-inflammatory cytokines to the total amount of pro-inflammatory cytokines produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials. In some embodiments, under otherwise identical conditions, the ratio of the total amount of anti-inflammatory cytokines (e.g., IL-10) produced per macrophage to the total amount of pro-inflammatory cytokines (e.g., IL-1β) produced in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein is at most 30 times greater than the ratio of the total amount of anti-inflammatory cytokines to the total amount of pro-inflammatory cytokines produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials. In some embodiments, under otherwise identical conditions, the ratio of the total amount of anti-inflammatory cytokines (e.g., IL-10) produced per macrophage to the total amount of pro-inflammatory cytokines (e.g., IL-1β) produced in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein is at most 50 times greater than the ratio of the total amount of anti-inflammatory cytokines to the total amount of pro-inflammatory cytokines produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials. In some embodiments, under otherwise identical conditions, the ratio of the total amount of anti-inflammatory cytokines (e.g., IL-10) to the total amount of pro-inflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing an immunomodulatory compound and / or material described herein is at most 75 times the ratio of the total amount of anti-inflammatory cytokines to the total amount of pro-inflammatory cytokines produced per macrophage in the presence of a macroencapsulated device without an immunomodulatory compound and / or material.In some embodiments, under otherwise identical conditions, the total amount of anti-inflammatory cytokines (e.g., IL-10) generated by each macrophage in the presence of a large encapsulated device containing immunomodulatory compounds and / or materials described herein to the total amount of pro-inflammatory cytokines (e.g., IL-1β) is at most 100 times the total amount of anti-inflammatory cytokines generated by each macrophage in the presence of a large encapsulated device without immunomodulatory compounds and / or materials to the total amount of pro-inflammatory cytokines. Combinations of the foregoing ranges are possible (e.g., at least 1.05 times and at most 100 times, at least 1.1 times and at most 30 times, at least 1.1 times and at most 40 times, at least 2 times and at most 25 times, or at least 6 times and at most 20 times). Other ranges are also possible.

[0079] The following methods can be used to measure the total amount of anti-inflammatory cytokines (for example, IL-10) generated by macrophages and the ratio of the total amount of proinflammatory cytokines (for example, IL-1 β).Macrophages (for example, monocytes, M0 macrophages or M1 macrophages) can be used for 6 days of immunomodulatory compounds or material treatment as described herein, and compared with otherwise identical control samples without immunomodulatory compounds.For each in treated samples and controls, in order to determine the IL-1 β generated by each cell and the IL-10 generated by each cell, the amount of IL-1 β and IL-10 generated by the macrophages in treated samples and controls can be measured, for example, using ELISA, and normalized by the cell viability (such as viable cell number) in the corresponding processed and control samples measured respectively.The cell viability of M0 macrophages can be measured, for example, by using CellTiterGlo luminescent cell viability assay kit. Then, the ratio of the amount of IL-1β to the amount of IL-10 per macrophage of each of the treated samples and the control can be calculated by dividing the amount of IL-1β produced per cell by the amount of IL-10 produced per cell of the corresponding sample.

[0080] In some embodiments, the use of a macroencapsulated device containing an immunomodulatory compound and / or material as described herein can be associated with reduced production of proinflammatory cytokines (e.g., IL-1β) compared to a macroencapsulated device without an immunomodulatory compound and / or material. In some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing an immunomodulatory compound and / or material as described herein can be at least 10%, at least 20%, at least 30%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% less than the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device without an immunomodulatory compound and / or material as described herein. For example, in some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein can be at least 10% less than the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials described herein. In some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein can be at least 20% less than the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials described herein. In some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein can be at least 30% less than the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials described herein. In some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein can be at least 50% less than the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials described herein. In some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing an immunomodulatory compound and / or material described herein can be at least 60% less than the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device without an immunomodulatory compound and / or material described herein.In some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein can be at least 70% less than the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials described herein. In some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein can be at least 80% less than the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials described herein. In some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein can be at least 90% less than the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials described herein. In some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein can be at least 95% less than the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials described herein. In some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein can be up to 50%, up to 60%, up to 70%, up to 80%, up to 90%, up to 95%, or up to 99% less than the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials described herein. In some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein can be up to 50% less than the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials described herein.In some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein can be up to 60% less than the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials described herein. In some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein can be up to 70% less than the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials described herein. In some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein can be up to 80% less than the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials described herein. In some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing immunomodulatory compounds and / or materials described herein can be up to 90% less than the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device without immunomodulatory compounds and / or materials described herein. In some embodiments, in the presence of a large encapsulated device containing an immunomodulatory compound and / or material as described herein, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage can be up to 95% less than in the presence of a large encapsulated device without an immunomodulatory compound and / or material as described herein. In some embodiments, in the presence of a large encapsulated device containing an immunomodulatory compound and / or material as described herein, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage can be up to 99% less than in the presence of a large encapsulated device without an immunomodulatory compound and / or material as described herein. Combinations of the above ranges are possible (e.g., at least 10% and up to 99%, at least 20% and up to 95%, at least 30% and up to 80%, at least 50% and up to 99%, or at least 60% and up to 95%). Other ranges are also possible.For example, in some embodiments, the amount of proinflammatory cytokines (e.g., IL-1β) produced per macrophage in the presence of a macroencapsulated device containing an immunomodulatory compound and / or material described herein can be 10-99%, 10-95%, 10-90%, 10-80%, 10-70%, 10-60%, 10-50%, 10-30%, 10-20%, 20-99%, 20-95%, 20-30%, 20-40%, 20-50%, 20-60%, 20-70%, 20-80%, 20-99%, 20-95%, 20-30%, 20-30%, 20-40%, 20-5 ... 0-90%, 20-80%, 20-70%, 20-60%, 20-50%, 20-30%, 30-90%, 30-95%, 30-90%, 30-80%, 30-70%, 30-60%, 30-50%, 50-99%, 50-95%, 50-90%, 50-80%, 50-70%, 50-60%, 60-99%, 60-95%, 60-90%, 60-80%, 60-70%, 70-99%, 70-95%, 70-90%, 70-80%, 80-99%, 80-95% or 90-99%. The amount of proinflammatory cytokines produced per macrophage cell (eg, IL-1β per cell) can be determined using methods described elsewhere herein.

[0081] The macroencapsulated devices disclosed herein can use any suitable type of membrane and can have any suitable type of construction for containing a cell population to deliver a desired therapeutic compound to a subject. In addition to retaining the cell population within the interior of the device, in some embodiments, the membrane of the device can be configured to protect one or more cell populations housed within the interior of the device from immune attack while allowing the desired biological product produced by the cells, such as insulin, as well as waste and nutrients used and produced by the cells, to pass through the semi-permeable portion of the membrane. In some embodiments, the membrane is configured to protect the cells from immune attack in the absence of immunosuppressive therapy.

[0082] The membrane of the macroencapsulated device can be formed from any suitable biocompatible material. The biocompatible material can be substantially inert to the cells contained within the macroencapsulated device and the surrounding tissue. The biocompatible material can comprise a synthetic polymer or a naturally occurring polymer. In some embodiments, the polymer can also be a linear polymer, a cross-linked polymer, a network polymer, an addition polymer, a condensation polymer, an elastomer, a fibrous polymer, a thermoplastic polymer, a non-degradable polymer, combinations of the foregoing, and / or any other suitable type of polymer, as the present disclosure is not limited in this manner. Suitable types of polymers may include polyvinyl chloride (PVC), polyethylene (PE), polypropylene (PP), polymethyl methacrylate (PMMA), polystyrene (PS), polytetrafluoroethylene (PTFE), expanded polytetrafluoroethylene (ePTFE), polyurethane (PU), polyamide (nylon), polyethylene terephthalate (PET), polyethersulfone (PES), polyetherimide (PEI), polyvinylidene fluoride (PVDF), polycaprolactone (PCL), poly (lactic acid-co-glycolic acid) (PLGA), poly-L-lactide (PLLA), any combination of the foregoing and / or any other suitable polymeric material. The synthesis method for forming one or more porous membranes in the porous membrane from the above-mentioned polymeric materials may include, but is not limited to, expansion, solvent casting, immersion precipitation and phase separation, electrospinning, methods for generating iso-reticular networks, methods for generating trabecular networks, or any other suitable methods for forming porous polymeric membranes.

[0083] The sintering of the membrane can be used to change the porosity and flux characteristics of the membrane. For example, sintering can increase the porosity of the membrane while maintaining its pore structure. Sintering can also improve the mechanical stability and diffusion flux of the membrane. Therefore, sintering can be used to change the porosity and / or mechanical properties of the membrane, which in turn can be used to adjust the porosity and flux characteristics of the large encapsulated device. Therefore, in some embodiments, any desired combination of sintered membranes and / or unsintered membranes can be used. For example, the two outer membranes of the device can be bonded together, wherein the sintered membrane and the unsintered membrane are bonded together, two sintered membranes are bonded together, or two unsintered membranes are bonded together. In addition, in the case where these intermediate membranes can be sintered or unsintered, any number of intermediate membranes positioned between these outer membranes can be used.

[0084] The membrane of macroencapsulated device as described herein can be made of porous membrane material, and described porous membrane material is configured to allow by film transport such as treatment or biological product, and it has the molecular weight of less than about 3000kDa, 2000kDa, 1000kDa, 500kDa, 400kDa, 300kDa, 200kDa, 100kDa, 50kDa, 40kDa, 30kDa, 20kDa, 10kDa, 6kDa, 5kDa, 4kDa, 3kDa, 2kDa, 1kDa and / or any other suitable range, depending on the desired application. Combinations of the above ranges are possible (e.g., at least 1kDa and less than or equal to 3000kDa). Other ranges are also possible. The film of large encapsulated device as described herein can be made of porous membrane material, described porous membrane material is configured to allow by film transport only the material in the molecular weight range of 1-3000kDa, 1-2000kDa, 1-1000kDa, 1-500kDa, 1-400kDa, 1-300kDa, 1-200kDa, 1-100kDa, 1-50kDa, 1-40kDa, 1-30kDa, 1-20kDa, 1-10kDa, 1-6kDa, 1-5kDa, 1-4kDa, 1-3kDa or 1-2kDa, as bioproduct.For example, one or more films of large encapsulated device can be configured to allow the insulin that molecular weight is about 5.8kDa to flow through film.In certain embodiments, one or more films of large encapsulated device can be configured to allow only the material in the scope of 1-10kDa as bioproduct to flow. In certain embodiments, one or more membranes of large encapsulated device can be configured to allow material such as biological product only in the scope of 1-6kDa to flow. In certain embodiments, one or more membranes of large encapsulated device can be configured to allow material such as biological product only in the scope of 1-5kDa to flow. In certain embodiments, one or more membranes of large encapsulated device can be configured to allow material such as biological product only in the scope of 1-4kDa to flow. In certain embodiments, one or more membranes of large encapsulated device can be configured to allow material such as biological product only in the scope of 1-3kDa to flow. In certain embodiments, one or more membranes of large encapsulated device can be configured to allow material such as biological product only in the scope of 1-2kDa to flow. For example, one or more membranes of large encapsulated device can be configured to allow molecular weight to be about 5.8kDa insulin to flow through film. In addition, the size of the pore of film can be set to allow one or more immunomodulatory compounds to transport by material membrane.

[0085] To provide the desired selectivity, the porous membranes used with the macroencapsulation devices disclosed herein can have an open porous structure with an average pore size greater than or equal to about 1 nm, 5 nm, 10 nm, 15 nm, 20 nm, 30 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm, 200 nm, 300 nm, and / or any other suitable size range. Correspondingly, the average pore size of the various films described herein can have an average pore size less than or equal to 2500nm, 2000nm, 1700nm, 1500nm, 1400nm, 1300nm, 1200nm, 1100nm, 1000nm, 900nm, 800nm, 700nm, 600nm, 500nm, 400nm, 300nm, 200nm, 100nm, 90nm, 80nm, 70nm, 60nm, 50nm, 40nm, 30nm, 20nm and / or any other suitable size range. Contemplate the foregoing combination, including, for example, an average pore size between or equal to: 1nm and 20nm, 1nm and 2500nm, 50nm and 1200nm and / or any other suitable combination. In some embodiments, the average pore size of the various films described herein is between 25nm and 1500nm. In some embodiments, the average pore size of the various membranes described herein is between 50nm and 1200nm. In some embodiments, the average pore size of the various membranes described herein is between 50nm and 1000nm. In some embodiments, the average pore size has an upper limit of 1500nm. In some embodiments, the average pore size has an upper limit of 1200nm. In some embodiments, the average pore size has a lower limit of 25nm. In some embodiments, the average pore size has a lower limit of about 50nm. Although specific average pore sizes are described above, it should be understood that any appropriate average pore size can be used for the various membranes described herein, including average pore sizes greater than and less than the above-mentioned average pore sizes.

[0086] In order to provide sufficient strength and / or rigidity to the macroencapsulated device, the various membranes and frames can be made of sufficiently hard materials. The desired rigidity can be provided by an appropriate combination of Young's modulus, thickness, and overall construction of the material balanced with the desired permeability of the device. An appropriate Young's modulus for the various membranes and frames described herein can be at least 10 5 Pa, 10 6 Pa, 10 7 Pa, 10 8 Pa, 10 9 Pa, 10 10Pa and / or any other suitable modulus greater than and less than these ranges. Of course, ranges between the foregoing Young's moduli are contemplated, including, for example, a Young's modulus between or equal to about 10 6 Pa and 10 10 Pa.

[0087] In certain embodiments, it may be desirable that one or more membranes included in the membrane of the macroencapsulated device be hydrophilic, to promote cell loading into the macroencapsulated device and / or promote one or more fluids, biocompounds, therapeutic agents, cell nutrients, cell waste and / or other materials to flow through the membrane of the device. In addition, the hydrophilic outer membrane can also reduce the generation of fibrosis when the device is positioned in vivo. Therefore, the membrane of the macroencapsulated device can be made of hydrophilic material and / or treated with a hydrophilic coating. Suitable hydrophilic material can include but is not limited to suitable hydrophilic polymers, polyethylene glycol, polyvinyl alcohol, polydopamine, any combination thereof and / or can form a coating on the film or can be made into any other suitable hydrophilic material of the film.

[0088] The films described in the various embodiments of the macroencapsulated devices described herein can be bonded to each other using any appropriate bonding method, as the present disclosure is not limited in this manner. For example, adjacent films can be bonded to each other using adhesives, epoxy resins, welding or other fusion-based technologies (e.g., ultrasonic bonding, laser bonding, physical bonding, thermal bonding, etc.), mechanical clamping using a frame or fixture, and / or any other appropriate bonding method. In a specific embodiment, adjacent films can be bonded using a heating tool that is used to press or impact two or more films against each other with a predetermined pressure and / or force for a set fusion time. In view of the foregoing, it will be understood that the present disclosure is not limited to the use of any particular method for bonding films together.

[0089] In certain embodiments, it may be desirable to limit the maximum thickness of a large encapsulated device in a direction perpendicular to the plane where the maximum lateral dimension of the device is located. Therefore, one or more internal portions of the first and second films placed in the framework can be bonded together to limit the degree to which the films can be displaced relative to each other. These bonding portions of the film can be evenly dispersed in the internal portion of the film positioned in the framework. These bonding portions can have any appropriate shape, including, for example, a point, a line, a curve, or any other appropriate shape. Although the bonding internal portion can have any appropriate size for the desired application, in one embodiment, a bonding point is used, and the diameter of the bonding point can be greater than or equal to approximately 0.5mm, 0.75mm, 1mm, 1.25mm, 1.5mm, and / or any other appropriate diameter. Correspondingly, the diameter of the point can be less than or equal to approximately 3mm, 2.75mm, 2.5mm, 2.25mm, 2.0mm, and / or any other appropriate diameter. Combinations of the above-mentioned ranges are contemplated, including, for example, a diameter between or equal to 0.5mm and 3mm. While specific shape and size ranges are provided above, it should be understood that other shapes and sizes, both smaller and larger than those recited, are contemplated, as the present disclosure is not limited in this manner.

[0090] In some embodiments, it may be desirable to improve the vascularization of large encapsulated devices. Thus, in certain embodiments, one or more through holes can be formed in the one or more bonding portions, which are located in an interior portion of the membrane disposed radially inward from the frame of the device. In addition to growing around the upper and lower surfaces of the device, these through holes can also allow vasculature to grow through the through holes. The one or more through holes can be formed in the bonding portion of the membrane using laser ablation, mechanical puncture, cutting, or any other suitable method for forming through holes in the one or more bonding portions of the membrane.

[0091] In some embodiments, the internal volume formed between the first membrane layer and the second membrane layer comprises a plurality of channels. As described in detail below, in some embodiments, one or more portions of adjacent membranes can be bonded together so that the internal volume within the device is subdivided into a plurality of interconnected channels. In some embodiments, the plurality of interconnected channels can be shaped like a lumen, but any suitable shape or configuration of channels can also be used. The channels can have an internal maximum transverse dimension, such as an inner diameter, greater than or equal to 40 μm, 50 μm, 100 μm, 200 μm, 300 μm, 400 μm, and / or any other suitable size. Correspondingly, the channels can have an internal maximum transverse dimension less than or equal to 800 μm, 700 μm, 600 μm, 500 μm, 400 μm, and / or any other suitable size. Combinations of the foregoing are contemplated, including, for example, a plurality of channels having an internal maximum lateral dimension between or equal to: 40 μm and 800 μm, 40 μm and 100 μm, 100 μm and 200 μm, 200 μm and 400 μm, 400 μm and 500 μm, 500 μm and 600 μm, 600 μm and 800 μm, etc. Furthermore, the density of the interconnecting channels forming the various compartments of the device can be greater than or equal to about 10 channels / cm in a lateral plane of the device. 2 , 15 channels / cm 2 , 20 channels / cm 2 , 25 channels / cm 2 , 30 channels / cm 2 , 35 channels / cm 2 , 40 channels / cm 2 , 45 channels / cm 2 , 50 channels / cm 2 , 60 channels / cm 2 , 70 channels / cm 2 , 80 channels / cm 2 , 90 channels / cm 2 , 100 channels / cm 2 , 110 channels / cm 2 , 120 channels / cm 2 , 130 channels / cm 2 , 140 channels / cm 2 , 150 channels / cm 2 , 175 channels / cm 2 or 200 channels / cm 2 Also contemplated are ranges extending between any of the aforementioned channel densities, including, for example, channel densities between or equal to about 10 channels / cm 2 and 200 channels / cm2 , 10 channels / cm 2 and 50 channels / cm 2 , 50 channels / cm 2 and 100 channels / cm 2 , 100 channels / cm 2 and 150 channels / cm 2 or 150 channels / cm 2 and 200 channels / cm 2 Although densities greater and less than the above ranges are also contemplated.

[0092] As described herein, the macroencapsulated device can have any suitable combination of internal volume, external dimensions and / or other suitable physical parameters. For example, the internal volume covered by the outer membrane of the macroencapsulated device can be between or equal to: 40 μL and 250 μL, 40 μL and 100 μL or 100 μL and 250 μL. The width or maximum lateral dimension of the macroencapsulated device can also be between about 20 mm and 80 mm. In addition, in order to provide the desired diffusion of oxygen into the interior of the macroencapsulated device to support the cells contained therein, the maximum oxygen diffusion distance from the outside of the device to the internal part of the device (including the cell colony) can be less than 50 μm, 100 μm, 150 μm, 200 μm, 250 μm, 300 μm, 350 μm, 400 μm, 450 μm or 500 μm. Correspondingly, the maximum thickness or dimension perpendicular to the maximum lateral dimension of the entire device and / or the internal volume located within the device can be less than 50μm, 100μm, 150μm, 200μm, 250μm, 300μm, 350μm, 400μm, 450μm or 500μm.

[0093] Furthermore, in some embodiments, the ratio of the surface area to volume of the device can be greater than or equal to about 20 cm -1 , 40cm -1 , 50cm -1 、60cm -1 , 80cm -1 , 100cm -1 , 120cm -1 , 150cm -1 , 200cm -1 , 300cm -1 , 400cm -1 , 500cm -1 , 600cm -1 , 700cm -1 , 800cm -1 , 900cm -1 or 1000cm -1In some embodiments, the device may have an external surface area to volume ratio between 25 cm -1 and 1250cm -1 In some embodiments, the ratio of the surface area to volume of the device can be between 50 cm -1 and 1000cm -1 In some embodiments, the ratio of the surface area to volume of the device can be between 100 cm -1 with 500cm -1 Ranges extending between any of the aforementioned values ​​of the various dimensions and parameters, as well as ranges greater than or equal to the aforementioned ranges, are also contemplated.

[0094] While specific dimensions, parameters, and relationships related to the macroencapsulated device and the materials from which it is made are described above, it should be understood that dimensions, parameters, and relationships greater than and less than the aforementioned dimensions, parameters, and relationships are contemplated, as the present disclosure is not limited in this manner. Thus, any suitable combination of size, configuration, material properties, and / or relative performance parameters may be used for the device, depending on the desired application.

[0095] In some embodiments, the macroencapsulated device can include at least one cell colony positioned within the internal volume of the device. For example, the cell colony can be positioned within the internal volume formed between two or more opposing adventitia layers of the device, wherein the outer edge of the internal volume can be defined by one or more bonding portions extending around the periphery of the membrane or other appropriate portions of the membrane. In such embodiments, at least the adventitia of the device can be configured to prevent one or more cell colonies from passing through the device. Thus, one or more cell colonies can be maintained within the internal volume of the device. Of course, while it is noted that two adventitia layers are used to form a single internal volume, it is also contemplated that multiple intermediate membranes positioned between the adventitia of the device and / or multiple unconnected internal volumes within the device are used.

[0096] As described above, in some embodiments, the methods and systems disclosed herein can be used to diffuse therapeutic compositions across at least one membrane layer of a large encapsulated device that at least partially encapsulates a cell colony. For example, when a large encapsulated device containing a cell colony is implanted into an implantation site, a therapeutic composition (e.g., a biological product) can be produced by the encapsulated cell colony. Therefore, the therapeutic composition can diffuse across the first membrane layer and / or the second membrane layer. The membrane layer described in more detail below can be configured to prevent the cell colony from passing through the membrane layer while allowing the therapeutic composition to diffuse across the membrane layer. The cell colony can be any cell in the various cells described elsewhere herein, such as cells that produce insulin. The therapeutic agent can be any therapeutic agent in the various therapeutic agents described herein, such as insulin. Depending on the molecular weight and / or membrane properties (e.g., pore size, thickness, etc.) of the therapeutic composition, the therapeutic composition can diffuse at any diffusion rate in various appropriate diffusion rates. In some embodiments, the sustained release of the therapeutic composition can be achieved using the device and methods described herein. For example, therapeutic composition can be released into the tissue at the implantation site in the time period of extension, such as at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 6 weeks or longer time and / or at most 52 weeks, at most 40 weeks, at most 30 weeks, at most 20 weeks or any other suitable time period. The combination of the above-mentioned scope is possible (for example, at least 1 week and at most 52 weeks). In certain embodiments, therapeutic composition can be released into the tissue at the implantation site in at least 2 weeks. In certain embodiments, therapeutic composition can be released into the tissue at the implantation site in at least 4 weeks. In certain embodiments, therapeutic composition can be released into the tissue at the implantation site in at least 6 weeks. In certain embodiments, therapeutic composition can be released into the tissue at the implantation site, continues to exceed 52 weeks. Other scopes are also possible.

[0097] In certain embodiments, the cell colony contained in the internal volume of the large encapsulated device can be a cell colony that secretes insulin. In certain embodiments, the cell colony includes at least one cell derived from a stem cell-derived cell. In certain embodiments, at least one cell is a genetically modified cell. In some cases, compared with comparable cells that are not genetically engineered, at least one cell is genetically engineered to reduce the immune response of the subject when the device is implanted. In certain embodiments, the cell colony is a stem cell-derived cell with glucose-stimulated insulin secretion (GSIS) ability. For example, a suitable cell colony can include pancreatic progenitor cells, endocrine cells, beta cells, alpha cells, delta cells, one or more of the matrix or any combination thereof. In addition, the matrix can include isolated islet cells, isolated pancreatic cells, isolated tissue cells, stem cells, stem cell-derived cells, induced pluripotent cells, differentiated cells, transformed cells or expression systems that can synthesize one or more bio-products. Optionally, in certain embodiments, the matrix can include cells of the second type of the first type of cells that support the synthesis of one or more bio-products. In certain embodiments, the cell can be encapsulated before being placed in the matrix. In such embodiments, the cell can be encapsulated in a microcapsule or can be conformally coated. However, naked cells, ie, unencapsulated cells, may also be used. Of course, other types of cells may be used with the disclosed immunomodulatory compounds and macroencapsulation devices disclosed herein, as the present disclosure is not limited to any particular cell type.

[0098] In some embodiments, the present disclosure provides a composition comprising a cell population disclosed herein and any one or more immunomodulatory compounds. In some embodiments, at least 30% of the cells in the cell population are NKX6.1 positive and ISL1 positive. In some embodiments, at least 20% of the cells in the cell population are NKX6.1 negative and ISL1 positive. In some embodiments, less than 25% of the cells in the cell population are NKX6.1 positive and ISL1 negative. In some embodiments, less than 10% of the cells in the population are NKX6.1 negative and ISL1 negative. In some embodiments, 20-50%, 20-45%, 20-40%, 20-35%, 20-30%, 20-25%, 25-50%, 25-45%, 25-40%, 25-35%, 25-30%, 30-50%, 30-45%, 30-40%, 30-35%, 35-50%, 35-45%, 35-40%, 40-50%, 40-45% or 45-50% of the cells in a population are NKX6.1 positive, ISL1 positive. In some embodiments, 15-50%, 15-40%, 15-30%, 15-25%, 20-50%, 20-45%, 20-40%, 20-35%, 20-30%, 20-25%, 25-50%, 25-45%, 25-40%, 25-35%, 25-30%, 30-50%, 30-45%, 30-40%, 30-35%, 35-50%, 35-45%, 35-40%, 40-50%, 40-45% or 45-50% of the cells in a population are NKX6.1 negative and ISL1 positive. In some embodiments, 1-25%, 1-15%, 1-8%, 2-25%, 2-15%, 2-8%, 3-25%, 3-15%, 3-8%, 6-25%, 6-15%, 6-10%, 9-25%, 9-15%, 15-25%, or 20-30% of the cells in a population are NKX6.1 positive and ISL1 negative. In some embodiments, 1-10%, 1-8%, 1-5%, 1-3%, 3-10%, 3-8%, 5-10%, 5-8%, 10-15%, or 0.5-1% of the cells in a population are NKX6.1 negative and ISL1 negative. In some embodiments, 20-50%, 20-45%, 20-40%, 20-35%, 20-30%, 20-25%, 25-50%, 25-45%, 25-40%, 25-35%, 25-30%, 30-50%, 30-45%, 30-40%, 30-35%, 35-50%, 35-45%, 35-40%, 40-50%, 40-45% or 45-50% of the cells in the population express insulin.In some embodiments, 15-50%, 15-40%, 15-30%, 15-25%, 20-50%, 20-45%, 20-40%, 20-35%, 20-30%, 20-25%, 25-50%, 25-45%, 25-40%, 25-35%, 25-30%, 30-50%, 30-45%, 30-40%, 30-35%, 35-50%, 35-45%, 35-40%, 40-50%, 40-45% or 45-50% of the cells in a population express glucagon.

[0099] In some embodiments, the present disclosure provides a composition comprising any one or more immunomodulatory compounds in a stem cell-derived β cell population and an immunomodulatory compound disclosed herein. The terms "stem cell-derived β cells", "SC-β cells", "functional β cells", "functional pancreatic β cells", "mature SC-β cells" and their grammatical equivalents may refer to cells (e.g., non-natural pancreatic β cells) that display at least one marker indicating pancreatic β cells (e.g., PDX-1 or NKX6.1) and express insulin. In some embodiments, SC-β cells display glucose-stimulated insulin secretion (GSIS) response characteristics of endogenous mature β cells. In some embodiments, the terms "SC-β cells" and "non-natural β cells" as used herein are interchangeable. In some embodiments, "SC-β cells" include mature pancreatic cells. It should be understood that SC-β cells do not need to be derived (e.g., directly) from stem cells. Examples of detailed protocols for generating endocrine cells from stem cells to provide at least one SC-islet cell (e.g., SC-β cell) are described in U.S. Patent Application Publication Nos. US20150240212, US20150218522, US20210198632, and US20220090020, PCT Publications WO2022 / 147056 and WO2022192300, and U.S. Patent No. 11,466,256, each of which is incorporated herein by reference in its entirety. Additional methods of making SC-β cells include, for example, U.S. Patent No. 10,030,229; U.S. Patent No. 10,443,042; U.S. Patent No. 11,466,256, published applications US20200332262, US20150240212, US20150218522, US20210198632, and US20220090020; and PCT publications WO2022 / 147056 and WO2022192300, each of which is incorporated by reference in its entirety.

[0100] In some embodiments, cells express one or more beta cell markers. The term "beta cell marker" refers to, but is not limited to, proteins, peptides, nucleic acids, polymorphisms of proteins and nucleic acids, splice variants, fragments of proteins or nucleic acids, elements, and other analytes specifically expressed or present in pancreatic beta cells. Exemplary beta cell markers may include, but are not limited to, pancreatic and duodenal homeobox 1 (PDX1) polypeptide, insulin, c-peptide, amylin, E-cadherin, Hnf3β, PCI / 3, B2, Nkx2.2, GLUT2, PC2, ZnT-8, ISL1, Pax6, Pax4, NeuroD, 1Inf1b, Hnf-6, Hnf-3β, and MafA, as well as Zhang et al., Diabetes. 50(10): Those described in 2231-6 (2001). In some embodiments, the beta cell marker is a nuclear beta cell marker. In some embodiments, the beta cell marker is PDX1 or PH3. In some embodiments, the cell population comprises non-naturally occurring cells that express C-peptide and ISL1.

[0101] In some embodiments, SC-β cells exhibit responses to multiple glucose loads (e.g., at least one, at least two, or at least three or more continuous glucose loads). In some embodiments, the response is similar to the response of endogenous islets (e.g., human islets) to multiple glucose loads. In some embodiments, the morphology of SC-β cells is similar to the morphology of endogenous β cells. In some embodiments, SC-β cells exhibit in vitro GSIS responses similar to the GSIS responses of endogenous β cells. In some embodiments, SC-β cells exhibit in vivo GSIS responses similar to the GSIS responses of endogenous β cells. In some embodiments, SC-β cells exhibit in vitro and in vivo GSIS responses similar to the GSIS responses of endogenous β cells. In some embodiments, the GSIS responses of a composition comprising any one or more immunomodulatory compounds of a stem cell-derived β cell population and an immunomodulatory compound disclosed herein can be observed within two weeks of transplanting SC-β cells into a host (e.g., a human or animal). In some embodiments, SC-β cells package insulin into secretory granules. In some embodiments, SC-β cells exhibit encapsulated crystalline insulin granules. In some embodiments, SC-β cells exhibit cytokine-induced apoptosis in response to cytokines. In some embodiments, in response to known antidiabetic drugs (e.g., secretagogues), insulin secretion from SC-β cells is enhanced. In some embodiments, SC-β cells are monohormonal. In some embodiments, SC-β cells do not abnormally co-express other hormones, such as glucagon, somatostatin, or pancreatic polypeptide. In some embodiments, SC-β cells exhibit a low replication rate. In some embodiments, SC-β cells increase intracellular Ca2+ in response to glucose. In some embodiments, SC-β cells express lower levels of MAFA compared to β cells from the pancreas of healthy control adult subjects. In some embodiments, SC-β cells express higher levels of MAFB compared to β cells from the pancreas of healthy control adult subjects. In some embodiments, SC-β cells express higher levels of SIX2, HOPX, IAPP, and / or UCN3 compared to β cells from the pancreas of healthy control adult subjects. In some embodiments, SC-β cells do not express MAFA. In some embodiments, SC-β cells express MAFB. In some embodiments, any of the cell markers disclosed herein (e.g., MAFA, MAFB, SIX2, HOPX, IAPP, and / or UCN3) are detected by flow cytometry.

[0102] In some embodiments, any cell in the cells disclosed herein comprises genome disruption in at least one gene sequence, wherein the disruption reduces or eliminates the expression of a protein encoded by the gene sequence. In some embodiments, the at least one gene sequence is an ABO sequence, such that the disruption causes the cell to be type O blood. In some embodiments, the at least one gene sequence encodes an MHC class I gene. In some embodiments, the MHC class I gene encodes beta-2 microglobulin (B2M), HLA-A, HLA-B, or HLA-C. In some embodiments, the at least one gene sequence encodes CIITA. In some embodiments, the cell comprises genome disruption in genes encoding HLA-A and HLA-B, but does not comprise genome disruption in genes encoding HLA-C. In some embodiments, the cell comprises genome disruption in a gene encoding CXCL10. In some embodiments, the cell comprises genome disruption in a gene encoding renal enzyme. In some embodiments, the cell comprises genome disruption in a natural killer cell that activates a ligand gene. In some embodiments, the natural killer cells activating ligand genes encode intercellular adhesion molecule 1 (ICAM1), CD58, CD155, carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1), cell adhesion molecule 1 (CADM1), MHC class I polypeptide-related sequence A (MICA), or MHC class I polypeptide-related sequence B (MICB). In some embodiments, the cells have decreased expression of one or more of beta-2 microglobulin, CIITA, HLA-A, HLA-B, HLA-C, HLA-DP, HLA-DQ, and HLADR relative to non-genetically modified cells. In some embodiments, relative to non-genetically modified cells, cells have increased CD47, PDL1, HLA-G, CD46, CD55, CD59, CTLA, PDL2, HLA-C, HLA-E, HLA-G, C1-inhibitor, IL-35, DUX4, IDO1, IL10, CCL21, CCL22, CD16, CD52, H2-M3, CD200, FASLG, MFGE8 and / or SERPINB9 expression. In a specific embodiment, compared to endogenous islet cells from healthy control subjects, cells disclosed herein (e.g., NKX6.1 positive, ISL1 positive cells) have increased PDL1 expression. In a specific embodiment, compared to endogenous islet cells from healthy control subjects, islet cells disclosed herein (e.g., SC-β cells) have increased CD47 expression. In some embodiments, genome disruption is induced by using a gene editing system, such as CRISPR Cas technology.In some embodiments, any of the isolated cells described herein (e.g., NKX6.1-positive, ISL1-positive cells) comprises a disruption (e.g., a deletion, insertion, translocation, inversion, or substitution of one or more nucleotides) in any one or more of the genes encoding B2M, CIITA, CXCL10, renalase, HLA-A, HLA-B, HLA-C, RFX-ANK, NFY-A, NLRC5, RFX5, RFX-AP, HLA-G, HLA-E, NFY-B, PD-L1, NFY-C, IRF1, TAPI, GITR, 4-1BB, CD28, B , PD-L2, CD58, CD2, HELIOS, IDO1, TRAC, TRB, NFY-A, CCR5, F3, CD142, MICA, MICB, LRP1, HMGB1, ABO, RHD, FUT1, KDM5D, PDGFRa, OLIG2, and / or GFAP. In some embodiments, disruption of a gene results in a decrease in expression of the gene by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% compared to the expression of the gene in cells of the same type without the disruption. In some embodiments, a gene is disrupted using CRISPR / Cas, piggybac transposons, TALENs, and / or zinc finger technology. In some embodiments, the gene is heterozygously disrupted in the cell. In other embodiments, the gene is homozygously disrupted in the cell.

[0103] In some embodiments, the cells described herein (e.g., isolated stem cells or NKX6.1-positive, ISL1-positive cells) are negative for A antigen and negative for B antigen. In some embodiments, the cells described herein are negative for A antigen. In some embodiments, the cells described herein are negative for B antigen. In some embodiments, the cells described herein (e.g., NKX6.1-positive, ISL1-positive cells) are negative for Rh antigen. In some embodiments, the cells described herein (e.g., isolated stem cells or NKX6.1-positive, ISL1-positive cells) are negative for A antigen, negative for B antigen, and negative for Rh antigen. As used herein, "A antigen" refers to a tissue-blood group antigen produced and expressed as a cell surface antigen by 3α-N-acetylgalactosaminyltransferase. As used herein, "B antigen" refers to a tissue-blood group antigen produced and expressed as a cell surface antigen by 3α-galactosaminyltransferase. In some embodiments, the cell comprises a disruption in the ABO gene. In some embodiments, the cell comprises a disruption in the ABO gene such that the cell has reduced or absent levels of A and B antigens. In some embodiments, the cells comprise a disruption in the FUT1 gene. In some embodiments, the cells comprise a disruption in the FUT1 gene such that expression of galactoside 2-α-L-fucosyltransferase 1 is reduced or absent. As used herein, "Rh antigens" refer to highly immunogenic antigens encoded by two highly polymorphic genes, RHD and RHCE. Rh antigen proteins are transmembrane proteins. In some embodiments, the cells comprise a disruption in the RHAG gene. In some embodiments, the cells comprise a disruption in the RHAG gene such that the cells have reduced or absent levels of Rh-associated glycoproteins. In some embodiments, the cells have reduced or eliminated expression of Rh protein antigens selected from the group consisting of: Rh C antigen, Rh E antigen, Kell K antigen (KEL), Duffy (FY) Fyα antigen, Duffy Fy3 antigen, Kidd (JK) Jkb antigen, MNS antigen U, and MNS antigen S.

[0104] In some embodiments, any of the cells disclosed herein (e.g., NKX6.1 positive, ISL1 positive cells) comprises a "safety switch". In some embodiments, the safety switch is a nucleic acid construct encoding a switch protein that inductively causes cell death or stops cell proliferation. In some embodiments, the safety switch is inserted into a defined specific target locus (e.g., a safe harbor locus) in the genome of the engineered cell, typically at two alleles of the target locus. In some embodiments, the target locus is a safe harbor locus, such as ActB or CLYBL. In some embodiments, the target locus is a gene targeted for destruction (e.g., B2M or CIITA). In some embodiments, the switch protein is activated by contact with an effective dose of a clinically acceptable orthologous small molecule. In some embodiments, when activated in some embodiments, the safety switch stops the cell from proliferating by activating apoptosis of the cell. In some embodiments, the switch protein comprises herpes simplex-thymidine kinase. In certain embodiments, the switch protein comprises a human caspase protein, such as caspase 1, caspase 2, caspase 3, caspase 4, caspase 5, caspase 6, caspase 7, caspase 8, caspase 9, caspase 10, caspase 14, etc. In certain embodiments, the protein is human caspase 9. In certain embodiments, the caspase protein is fused to a sequence that provides chemically induced dimerization (CID), wherein dimerization occurs only in the presence of an orthologous activator. One or more CID domains can be fused to the caspase protein, for example, two different CID domains can be fused to the caspase protein. In certain embodiments, the CID domain is a dimerization domain of an FKBP or FRB (FKBP-rapamycin binding) domain of an mTOR activated with a rapamycin analog. In some embodiments, the safety switch is any of the safety switches described in WO2021173449 and Jones et al., 2014, Frontiers in Pharmacology, 5(254): 1-8, each of which is incorporated herein in its entirety.

[0105] In some embodiments, the population further comprises a culture medium. In some embodiments, the culture medium comprises a sugar. In some embodiments, the sugar is sucrose or glucose. In some embodiments, the culture medium comprises a sugar at a concentration between about 0.05% and about 1.5%. In some embodiments, the culture medium is CMRL culture medium; or wherein the culture medium is FRS storage medium.

[0106] Depending on the specific embodiment, cells of therapeutically effective density can be loaded into one or more compartments of the macroencapsulated device. Suitable cell densities placed in the compartments can be greater than or equal to about 1,000 cells / μL, 10,000 cells / μL, 50,000 cells / μL, 100,000 cells / μL, 500,000 cells / μL, 750,000 cells / μL, 1,000,000 cells / μL and / or any other suitable cell density. Suitable cell densities placed in the compartments can also be less than or equal to about 1,000,000 cells / μL, 500,000 cells / μL, 100,000 cells / μL, 50,000 cells / μL, 10,000 cells / μL and / or any other suitable cell density. The foregoing combinations are contemplated, including cell densities between about 1000 cells / μL and 1,000,000 cells / μL. In some embodiments, the cell density placed in the compartment is between 100,000 cells / μL and 1,000,000 cells / μL. In some embodiments, the cell density placed in the compartment is between 75,000 cells / μL and 500,000 cells / μL. In some embodiments, the cell density placed in the compartment is between 500,000 cells / μL and 1,000,000 cells / μL. In some embodiments, the cell density placed in the compartment is between 750,000 cells / μL and 1,000,000 cells / μL. In some embodiments, the cell density placed in the compartment is between 750,000 cells / μL and 1,250,000 cells / μL. Of course, cell densities greater and less than the aforementioned cell densities can also be used, depending on the desired application and the cell type being used.

[0107] The macroencapsulated devices described herein can be implanted in a subject at various locations within the body. In one example, the device can be implanted in a subject via preperitoneal or retrorectus implantation. In other examples, the device can be placed via intra-omental implantation. In another example, the device can be placed via subcutaneous implantation. In another example, the device can be placed via suprahepatic implantation. In some cases, the macroencapsulated devices described herein can be secured at the implantation site within the body using any suitable fixation method, including, for example, applying a tissue adhesive. Suitable tissue adhesives can include, but are not limited to, fibrin, cyanoacrylates, polyethylene glycol, albumin-based adhesives, polymer-based adhesives, and / or any other suitable adhesive. In another example, the device can be secured using platelet-rich plasma and / or any other suitable fixation method, as the present disclosure is not limited in this manner.

[0108] Turning to the drawings, specific non-limiting embodiments are described in further detail. It should be understood that the various systems, components, features, and methods described with respect to these embodiments may be used alone and / or in any desired combination, as the present disclosure is not limited to the specific embodiments described herein.

[0109] Figures 1A-1B Depicted is an embodiment of a large encapsulation device after the membrane has been mounted to a corresponding frame and before being filled with a desired material, such as a cell colony. Specifically, as shown in the figure, the large encapsulation device can include a first membrane 102, a second membrane 104, and a frame 220, which extends along at least a portion of the periphery of the first and second membranes. The first and second membranes can correspond to adjacent layers of a single membrane that has been folded onto itself, and / or the first and second membranes can be separate membranes, as the present disclosure is not limited to this approach. The device is shown in an unfilled, relaxed state, wherein the additional surface area of ​​the first and second membranes relative to the cross-sectional area of ​​the frame in which the membrane is mounted causes the adhesive film to be suspended below the frame due to the resulting relaxation in the membrane. Due to the adhesive portion 122, through-hole 132, and other appropriate features already formed on the membrane within the internal area of ​​the device, the large encapsulation device can now be easily filled with a desired material, such as a cell colony, with minimal additional processing and handling. The internal volume can be filled using ports, openings in the peripheral adhesive portion, and / or any other appropriate method. In either case, after filling the macroencapsulated device with the desired material, the internal volume contained between the first membrane 102 and the second membrane 104 can expand, which can take up slack in the membranes because the membranes are placed under tension in the filled configuration due to the expansion of the internal volume between the membranes. This can cause the first and second membranes to deform so that the membranes extend generally in a direction generally parallel to the plane of the frame 220, see Figure 1C Correspondingly, due to this increase in the internal volume of the now filled device, the first and second membranes can now extend outwardly from opposing surfaces of the frame by approximately equal distances. With the membrane portions 132 having been bonded together at locations located radially inward from the frame, the expanded structure can again form a plurality of interconnected channels 126.

[0110] Any suitable filling method can be used to fill the macroencapsulated device. For example, a cell colony or other desired material can be flowed into the internal volume of the macroencapsulated device formed between the outer membranes of the device. This can be achieved by using a sealable or removable port extending into the internal volume, and / or openings can be present in the peripheral adhesive portion and / or frame of the macroencapsulated device, which can then be sealed. Although any suitable inlet to the internal volume can be used to allow material to flow into the internal volume of the device, the flow of this material can be controlled in a variety of different ways to provide the desired filling of the internal volume. For example, in one embodiment, the pressure applied to the internal volume of the macroencapsulated device can correspond to the desired tension present in the membrane of the device in the filling configuration. Thus, filling of the device can continue until a predetermined pressure and / or membrane tension threshold is reached. However, any suitable method for controlling the amount of material flowing into the internal volume can also be used, as the present disclosure is not limited to this method. This can include, for example, control based on the absolute volume of material flowing into the internal volume, the duration of a given flow rate, and / or any other suitable control method.

[0111] Figure 2 A diagram of a large encapsulated device filled with a cell population is shown. Similar to the above-described embodiment, the device can include a first membrane 102 and a second membrane 104 bonded along their perimeters to form an interior volume 250 between the membranes. The depicted device also includes a bonding portion 124, wherein a through-hole 132 is located within a central portion of the membranes positioned within a frame (not depicted). As shown, interior volume 250 comprises a plurality of interconnected channels.

[0112] Although specific embodiments of macroencapsulated devices have been illustrated in the figures, it should be understood that the various immunomodulatory compounds and materials disclosed herein may be used with any suitable type of macroencapsulated device, as the present disclosure is not limited in this regard.

[0113] Figure 3Shown is a flow chart of a method for delivering a therapeutic composition produced by a cell colony. When a large encapsulated device containing a cell colony is implanted into an implantation site, a therapeutic composition (e.g., a bioproduct) can be produced by the encapsulated cell colony. As shown in step 410, the therapeutic composition can diffuse into the surrounding tissue across the first membrane layer and / or the second membrane layer. Depending on the molecular weight and / or membrane properties (e.g., pore size, thickness, etc.) of the therapeutic composition, the therapeutic composition can diffuse with any diffusion rate in various appropriate diffusion rates. Before, during, or after the therapeutic composition diffuses across at least one membrane, the immunomodulatory compound can be released into the tissue around the large encapsulated device, as shown in step 420. In embodiments in which immunomodulatory material is placed on the surface of one or more membrane layers and / or in the pores, the immunomodulatory compound can be released from the surface and / or the pores of one or more membrane layers. Alternatively or in addition, in embodiments in which immunomodulatory material or compound is fixed to one or more membrane layers by one or more linker molecules, the immunomodulatory compound can be released from one or more membrane layers when one or more linker molecules are degraded. Alternatively or additionally, where the immunomodulatory material or compound is applied directly to the tissue surrounding the implantation area of ​​the one or more membrane layers, the immunomodulatory material or compound can be released upon deposition into the tissue. Next, as shown in step 430, upon release of the immunomodulatory material or compound into the tissue, the material or compound can be exposed to the surrounding tissue and induce a favorable foreign body response, such as a decrease in proinflammatory cytokines and / or an increase in the amount of anti-inflammatory cytokines relative to proinflammatory cytokines produced by macrophages.

[0114] Figure 4 A schematic cross-sectional view of a macroencapsulated device according to some embodiments is shown, comprising an interior volume and an immunomodulatory material associated with one or more membrane layers. In the depicted embodiment, the device 300 comprises a first outer membrane 102, a second outer membrane 104, and an interior volume corresponding to a formed interior compartment 250, wherein the first outer membrane and the second membrane are bonded to each other around the perimeter of the interior compartment. Figure 4As shown, one or more immunomodulatory compounds 145 can be placed in the device in any way in various appropriate ways. For example, the immunomodulatory compound can be placed on the surface (for example, outer surface) of one or more film layers 102 / 104 as a topcoat 170 or 180. In some cases, the immunomodulatory compound can be accommodated together with a matrix 160 (for example, a hydrogel matrix) and deposited on the film layer as a topcoat 180. Alternatively, the immunomodulatory compound can be deposited separately on the film layer (without matrix) as a topcoat 170. In some cases, the immunomodulatory material or compound can be fixed to one or more film layers 102 and / or 104 and maintained on a part of the device or in a part of the device by a linker molecule 150. Alternatively or in addition, the immunomodulatory material or compound included in the composition can be impregnated in the pores of the membrane (not depicted).

[0115] In another embodiment, the immunomodulatory material or compound can be placed in the internal volume 250 of the macroencapsulated device. For example, the immunomodulatory material or compound can be placed in the internal volume 250 alone or together with a matrix 160 (e.g., a polymer or hydrogel). In some cases, the immunomodulatory material or compound can be physically mixed with the matrix 160. In some such cases, the immunomodulatory material or compound can be released by diffusing out of the matrix 160. In other cases, the immunomodulatory material or compound can be fixed to the matrix 160. As previously described, in some embodiments, the matrix 160 can be a biodegradable material that can be used to release the immunomodulatory material or compound when the material degrades. It should be noted that different types of materials can be selected to adjust the degradation rate and adjust the release rate of the immunomodulatory material or compound.

[0116] Alternatively or additionally, the immunomodulatory material can be positioned adjacent to one or more membrane layers, such as outside and adjacent to the first membrane layer 102 and / or the second membrane layer 104. For example, the immunomodulatory material 190 containing the compound (in the matrix 160) can be applied to the tissue surrounding the macroencapsulated device such that the immunomodulatory material and / or compound is positioned around at least a portion of the macroencapsulated device.

[0117] Although not in Figure 4 , but the interior volume 250 can be configured to house a cell population and one or more immunomodulatory materials or compounds as indicated. Alternatively, the one or more immunomodulatory materials or compounds 145 can be fixed to the inner surface, outer surface and / or pores of the membrane layer enclosing the interior volume 140, which houses the cell population.

[0118] It should be noted that although Figure 4A macroencapsulated device comprising a single internal volume is shown, wherein one or more immunomodulatory materials or compounds are disposed within and / or affixed to a portion of the macroencapsulated device, but the disclosed methods, materials, and / or compounds may be used with any suitable type of macroencapsulated device, including any number of internal volumes, arrangement of one or more semipermeable membranes, or other suitable configuration. Figure 4 Any of the foregoing embodiments relating to one or more immunomodulatory materials or compounds shown in and described elsewhere herein are also applicable to any of the other devices and methods disclosed herein, as the present disclosure is not limited to use with any particular configuration of macroencapsulated devices.

[0119] Example 1

[0120] Macrophages play a key role in the foreign body response (FBR). Classically activated macrophages (M1) are pro-inflammatory, while alternatively, activated macrophages (M2) are anti-inflammatory. This example demonstrates that polarization of macrophages toward M2 macrophages can reduce the FBR and promote angiogenesis and vascularization. In this example, a panel of immunomodulatory compounds was tested to determine their effects on macrophage polarization, angiogenesis, and vascularization.

[0121] Human mononuclear cells (THP-1, TIB202 TM Naive macrophages (M0) were differentiated by incubating M0 macrophages with the compounds for 3 days. A panel of compounds was tested for their effects on macrophage polarization by incubating M0 macrophages with the compounds for 6 days. Secretion of interleukin (IL)-1β (a pro-inflammatory cytokine) and IL-10 (an anti-inflammatory cytokine) was measured using enzyme-linked immunosorbent assays (ELISAs), respectively.

[0122] Pro-inflammatory macrophages (M1) were differentiated by treating M0 macrophages with 100 ng / mL lipopolysaccharide (LPS) and 100 ng / mL interferon-γ (IFN-γ) for 3 days. A panel of compounds was tested for their effects on macrophage polarization by culturing M0 with the compounds for 6 days. The secretion of IL-1β (pro-inflammatory cytokine) and IL-10 (anti-inflammatory cytokine) was measured using enzyme-linked immunosorbent assays (ELISAs), respectively.

[0123] M0 macrophages were treated with various compounds for 6 days. These compounds cover a wide range of inhibitors for different cell signaling pathways. Figure 5As shown, the group of compounds tested includes compounds belonging to natural products (e.g., tannic acid, pentacyclic triterpenoid lupeol), CSF1R inhibitors (e.g., pegidatinib, edicotinib, GW2580, BLZ945), CCR2 inhibitors (e.g., PF-04136309), NF-kB inhibitors (e.g., sitagliptin), JAK inhibitors (e.g., ruxolitinib, baricitinib, tofacitinib), PDE4 inhibitors (e.g., roflumilast, apremilast), ROCK inhibitors (e.g., fasudil), NLRP3 inhibitors (e.g., glyburide, forskolin), metabolites (e.g., succinate, itaconate). Figure 5 As shown, the concentration of all test compounds was 1.0 μM except for Pesidatinib (0.25, 0.5 and 1.0 μM). For each treatment condition and drug-free control, the cell viability of M0 was measured by using the CellTiterGlo luminescent cell viability assay kit to show the cytotoxicity of the compound. The amount of IL-1β and IL-10 for each compound treatment and drug-free control was normalized by the corresponding cell viability measured for the corresponding treatment condition or control. The ratio of the normalized IL-10 amount to the normalized IL-1β amount was then calculated for each compound treatment and drug-free control ( Figure 5 The fold change relative to the no-drug control ratio was calculated by dividing the ratio for each compound treatment by the ratio for the no-drug control.

[0124] like Figure 5 As shown, various compounds exhibited elevated IL-10 to IL-1β ratios relative to the no-drug control. Specifically, for tannic acid and pegidatinib, a fold change in the ratio of at least 4 was observed. Tannic acid and pegidatinib reduced IL-1β secretion and increased IL-10 secretion, demonstrating effects on downregulation of M1 macrophages and upregulation of anti-inflammatory macrophages (M2). While tannic acid is a polyphenolic compound with redox scavenging and anti-inflammatory properties, pegidatinib (PLX-3391) is an inhibitor of the colony stimulating factor 1 receptor (CSF1R).

[0125] In addition, the cytotoxicity of the test compounds to macrophages was evaluated. The change fold of cell viability relative to the drug-free control was calculated by dividing the cell viability of each compound treatment by the cell viability of the drug-free control. Pecidinib and ruxolitinib at a concentration of 1.0 μM showed cytotoxicity. However, lower concentrations of pecidinib (0.25 and 0.5 μM) maintained cell viability at the same drug-free control level. All other compounds had no cytotoxicity to macrophages. In addition, tannic acid and pecidinib showed a high ratio of IL-10 to IL-1β. All these data indicate that tannic acid (1 μM) and pecidinib (0.25 and 0.5 μM) may be able to inhibit the pro-inflammatory response of M0 macrophages.

[0126] like Figure 6-7 As shown in Figure 2, M0 macrophages were treated with different compounds at different concentrations. These compounds included pesidatinib, tannic acid, gallic acid, rhein, curcumin, TAK242, and verteporfin. Figure 6 As shown, pecidinib (0.125 and 0.25 μM), tannic acid (0.25, 0.5, and 1.0 μM), and TAK242 (2.0 and 4.0 μM) reduced proinflammatory IL-1β production per cell, and the effects were dose-dependent. On the other hand, rhein (4.0 μM) and curcumin (1.0 μM) increased IL-1β levels. TAK242 is an inhibitor of toll-like receptor 4 (TLR4) and nuclear factor kappa light-chain-enhancer of activated B cells (NF-κB). In addition, as Figure 7 As shown in Figure 3, MCC950 (0.25 to 8 μM) reduced IL-1β levels, but the effect was not dose-dependent, suggesting that a concentration of 0.25 μM may have reached a threshold for the effect level. MCC950 is an inhibitor of the NOD, LRR, and pyrin domain-containing protein 3 (NLRP3) inflammasome. Figure 5-7 As shown, pecidinib (0.125 and 0.25 μM), tannic acid (0.25, 0.5 and 1.0 μM), TAK242 (2.0 and 4.0 μM) and MCC950 (0.25 to 8.0 μM) inhibited IL-1β secretion per cell, which may indicate a reduction in the pro-inflammatory response of macrophages. The molecular structures of the above compounds are provided in Figures 8A-8D In. Figures 8A-8D As shown, tannic acid and TAK242 are both toll-like receptor 4 (TLR4) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) inhibitors, MCC950 is an NLRP3 inflammasome inhibitor, and pegidatinib is a CSF1R inhibitor.

[0127] Example 2

[0128] In this example, a study was conducted to determine the effective in vivo dose of a group of immunomodulatory compounds that can reduce proinflammatory macrophage responses. First, a dose range that can inhibit proinflammatory macrophages was identified. Second, the identified dose range was evaluated to exclude any dose that (i) was cytotoxic to pancreatic islets (therapeutic cell products) and endothelial cells, and (ii) prevented in vitro endothelial network formation (which simulates angiogenesis and vascularization in vivo). Finally, the selected dose range was tested for drug loading and delivery.

[0129] Dose-dependence of various compounds on inhibition of proinflammatory responses

[0130] Figures 9A-9B is shown by M0 macrophages ( Figure 9A ) or M1 macrophages ( Figure 9B ) Graph showing the fold change in IL-1β secretion under each compound condition relative to the control (DMSO control without any compound). Figures 9A-9B As shown, a dose range of tannic acid, pesidatinib, TAK242, and MCC950 was identified that was able to inhibit the proinflammatory response of M0 and M1 macrophages. For M0 macrophages, tannic acid was effective at 0.25, 0.5, and 1.0 μM, pesidatinib was effective at 0.0625, 0.125, and 0.25 μM, TAK242 was effective at 2.0 and 4.0 μM, and MCC950 was effective at 0.25, 0.5, 1.0, 2.0, 4.0, and 8.0 μM ( Figure 9A For M1 macrophages, tannic acid was effective at 1.0 μM, pesiditinib was effective at 0.0625, 0.125, and 0.25 μM, TAK242 was effective at 4.0 μM, and MCC950 was effective at 0.25, 0.5, 1.0, 2.0, 4.0, and 8.0 μM.

[0131] like Figure 10 As shown, rat islets were purchased from the Joslin Diabetes Center and seeded into ultra-low 96-well plates (0.16 mL, 200 islets / mL, 32 islets per well). The fold change in cell viability relative to the drug-free control was calculated by dividing the cell viability of each compound-treated cell by the cell viability of the DMSO control (without any compound). Rat islets were treated with immunomodulatory compounds (pecitartinib, tannic acid, TAK242, and MCC950) for 4 days. The cytotoxicity of the compounds was evaluated using a 3D cell viability assay kit. The results showed that tannic acid (1.0 μM), pecidinib (0.25 μM), and MCC950 (8.0 μM) unexpectedly reduced the viability of rat pancreatic islets.

[0132] Cytotoxicity of various compounds on endothelial cells

[0133] like Figure 11 The cytotoxicity of the compounds to endothelial cells was evaluated on cultured monolayers of human umbilical vein endothelial cells (HUVECs). HUVECs were treated with different compounds at different concentrations for 6 days. Cell proliferation assay kit measures cell number. Compared to normal culture medium control (no compound treatment), a decrease in cell number may indicate the cytotoxicity of the compound. As shown, tannic acid and pesidatinib did not show cytotoxicity at the selected doses. MCC950 was observed to be associated with an increase in cell number. However, higher doses (4.0 and 8.0 μM) of TAK242 showed cytotoxicity to HUVEC.

[0134] Effects of various compounds on endothelial network formation

[0135] In addition, if Figures 12A-12Q As shown, the effect of compounds on endothelial network formation was evaluated by culturing HUVEC in collagen gel. HUVEC expressing green fluorescent protein (GFP) were embedded in collagen gel, and then compounds (e.g., 0.25μM-4.0μM MCC950, 1.0μM-8.0μM TAK242, 0.0625μM-0.25μM Pecidinib, 0.25μM–4.0μM tannic acid) were added to EGM-2 culture medium to study whether they would affect endothelial network formation in collagen gel. After 6 days of treatment, the endothelial network was imaged under a fluorescence microscope. Compared with the EGM-2 culture medium control ( Figure 12A ) and DMSO drug vehicle control ( Figure 12B ) in the network, the selected concentration of tannic acid ( Figures 12C-12E ), Pecidinib ( Figures 12F-12H ) and MCC950( Figures 12M-12Q ) showed a comparable level of endothelial network formation. However, 8.0 μM TAK242 appeared to inhibit endothelial network formation ( Figures 12I-12L ).

[0136] Another tubular endothelial network formation assay was performed using various concentrations of GDC-2394. For this assay, HUVECs were cultured on matrigel and exposed to GDC-2394 for a period of 24 hours. The data showed that tubular formation occurred with the various GDC-2394 concentrations tested (0.1 μM, 0.5 μM, and 2.5 μM). Figure 12R-12T ).

[0137] In summary, the dose ranges in which the compounds appeared both active (effective) and non-cytotoxic were identified. For tannic acid, the dose range was 0.25 μM to 0.5 μM. For pesidatinib, the dose range was 0.125 μM to 0.25 μM. For TAK242, the dose range was 2.0 μM to 4.0 μM. For MCC950, the dose range was 0.25 μM to 4.0 μM. For GDC-2394, the dose range was 0.1 μM to 2.5 μM. Once loaded into the delivery vehicle, the release of the compound can be controlled for approximately 10 days to suppress the pro-inflammatory response of macrophages.

[0138] Example 3

[0139] This example demonstrates various encapsulation schemes for various immunomodulatory compounds. Specifically, fibrin glue (FG) is used as an encapsulating agent for encapsulating immunomodulatory (IMM) compounds (e.g., tannic acid, MCC950, pegidatinib, and TAK242). The fibrin glue-encapsulated immunomodulatory compounds can be implanted as an injectable hydrogel or can be lyophilized to produce a sponge ( Figures 13A-13B ) for easy storage and transportation. Figures 13A-13B Shows the results before swelling with PBS ( Figure 13A ) and after swelling ( Figure 13B ) of freeze-dried fibrin glue encapsulated with immunomodulatory compounds.

[0140] To encapsulate the IMM compound into the fibrin glue, fibrinogen and thrombin were collected separately using a dual-chamber syringe provided by the manufacturer (Baxter Healthcare Corp). 4 μL of different concentrations of IMM compound in dimethyl sulfoxide (DMSO) were mixed with 21 μL of thrombin, and then 25 μL of fibrinogen was added to this mixture. The mixture was then incubated at 37°C for 30 minutes to form the IMM-encapsulated fibrin glue.

[0141] Fibrin glue containing different fibrinogen concentrations (10%, 25%, 50% and 100%) was used to successfully form a hydrogel within 30 minutes at 37°C. As shown, with higher fibrinogen (100%) (as Figure 14B Compared with the Figure 14A The fibrin glue construct (shown) had a larger pore size.

[0142] To study the release of IMM compounds from fibrin glue, IMM-encapsulated FG constructs were placed in 1.2 mL of cell culture medium containing 10% fetal bovine serum (FBS) and incubated at 37°C. At predetermined time points, the culture medium was collected for HPLC analysis and replaced with 1.2 mL of fresh culture medium. The amount of IMM compounds released from the culture medium was separated using a reversed-phase C18 column (Atlantis Silica T3) and detected using a PDA detector in the HPLC system.

[0143] Effect of fibrinogen concentration in FG on TA release

[0144] Fibrin glue (FG) constructs containing different fibrinogen concentrations (50% and 100%) and loaded with different doses of tannic acid (TA) (e.g., 2, 20, and 200 μg TA, respectively) were prepared to investigate the effect of fibrinogen concentration on the release profile of tannic acid from fibrin glue. Figure 15A ) of the fibrin glue (FG) construct provided a similar effect to that of 50% fibrinogen ( Figure 15B ). This can be explained by the smaller pore size of 100% FG compared to 50% FG, which may reduce the diffusion of immunomodulatory (IMM) molecules from the FG construct. A dose of 200 μg of tannic acid (TA) in 50 μL of 100% FG provided sustained release for up to 21 days. Based on these results, 100% fibrinogen was selected as the concentration for encapsulating other IMM compounds.

[0145] Effect of IMM dosage on the release of various IMMs from FG

[0146] Different doses of different immunomodulatory (IMM) compounds (MCC950, Pecidinib, and TAK242) were loaded into 50 μL fibrin glue (FG), and their release into the culture medium at 37°C was analyzed using HPLC ( Figures 16A-16C ). Higher doses of IMM compounds can be loaded into FG to prolong the release above the minimum effective concentration (MEC), as indicated by the dotted line. Figure 16A As shown, at a dose of 200 nmol, MCC950 was released from FG at a concentration above the MEC for 8 days. Figure 16C As shown in Figure 5, at a dose of 40 nmol, pecidinib was released continuously from FG for more than 10 days. Without wishing to be bound by any particular theory, the longer period of sustained release of pecidinib compared to MCC950 can be explained by its higher degree of hydrophobicity. Figure 10As shown in Figure 2, even if a high dose (e.g., 600 nmol) of TAK242 was loaded into FG, it was only able to maintain above the MEC for up to 2 days. Interestingly, on day 8, TAK242 released into the culture medium increased above the MEC, which may be due to the degradation of FG starting from this time point. Figure 16D As indicated, TAK242 was unstable and depleted more rapidly in cell culture medium at 37°C, whereas MCC950 and pecidinib were stable.

[0147] Effects of IMMs released from FG on macrophages

[0148] To investigate the effects of immunomodulatory (IMM) compounds released from fibrin glue (FG) on macrophages, an in vitro experiment was designed in which macrophages were in direct or indirect contact with FG. For indirect contact, M0 macrophages were seeded onto cell culture inserts with a pore size of 0.4 μm, while IMM-encapsulated FG was placed at the bottom of the pores ( Figure 17A For direct contact, IMM-encapsulated FGs were first placed on a cell culture insert, and then M0 macrophages were seeded on top of the construct ( Figure 17B ). Culture media were collected on days 4 and 7 for cytokine analysis (e.g., IL-1β, (TNF)-α, interferon (IFN)-γ). The results are shown in Figure 18 middle.

[0149] like Figure 18 As shown, IMM compounds were released from fibrin gels, and the effects of the released compounds on M0 macrophages were assessed. The fold change was calculated by dividing each compound treatment by the no-drug control. For IL-1β, at the low and high doses tested, pegstatinib did not reduce cytokine levels, low and high doses of MCC950 reduced cytokine secretion, and high doses of TAK242 showed an inhibitory effect ( Figure 18 ). For tumor necrosis factor (TNF)-α, pecidinib showed an inhibitory effect in the direct exposure group. MCC950 and TAK242 showed similar effects on IL-1β. The compounds' effects on interferon (IFN)-γ were similar to those on TNF-α. Overall, the direct exposure group generally appeared to be more effective than the indirect exposure group. High doses of MCC950 and TAK242 were the most effective, and the effects were able to last for 7 days.

[0150] Cytotoxicity of IMM compounds on SC-islets

[0151] To evaluate the cytotoxic effects of IMM compounds on human stem cell-derived islets (SC-islets), higher doses of the above compounds were loaded into fibrin glue (FG).

[0152] like Figure 19A As shown, the effects of the released compounds on the cell viability of SC-islets were evaluated. Only indirect groups were evaluated because there is no direct contact between SC-islets and fibrin glue in vivo. The fold change of dead cells was calculated by dividing each compound treatment by the drug-free control. As shown, a 12nmol dose of MCC950 increased the cell viability of SC-islets, while a 600nmol dose of TAK242 reduced the cell viability of SC-islets. Therefore, MCC950 appears to be effective in reducing proinflammatory cytokines and enhancing the viability of SC-islets. Consistent with SC-islets, the viability of porcine islets cultured with FG encapsulated with MCC950 was also significantly improved compared to the control without MCC950 ( Figure 19B ).

[0153] Figure 19C Shown are the percent viability of SC-islets after exposure to various concentrations of MD2-IN-1 or GDC-2394. The data show that the percent viability of SC-islets at various concentrations of the test compound evaluated was similar to the no drug control group.

[0154] In summary, it was found that various immunomodulatory compounds (e.g., tannic acid, peicidinib, TAK-242, MCC950) showed enhanced ability to reduce macrophage inflammatory responses while also providing low (if any) observed cytotoxicity, sustained release capacity, maintenance of vascularization, and / or other applicable benefits. For example, various immunomodulatory compounds (e.g., tannic acid, peicidinib, MCC950) showed the ability to prevent interruption of vascular network formation within a non-cytotoxic dose range and allowed sustained release over an extended period of time. In particular, TAK242 was observed to be effective in reducing proinflammatory cytokines without causing any observed toxicity to islets, and MCC950 appeared to be effective in reducing proinflammatory cytokines and enhancing islet viability while being non-cytotoxic to islets over a wide range of doses. Immunomodulatory compounds (e.g., peicidinib, MCC950) also showed drug stability over time in aqueous media at 37°C.

[0155] Example 4

[0156] This example demonstrates the release of immunomodulatory compounds (e.g., tannic acid (TA)) from sintered films relative to unsintered films. For example, various concentrations of tannic acid (10 μg / mL, 100 μg / mL, 1 mg / mL, 10 mg / mL) were used to soak sintered films ( Figure 20A ) and unsintered films ( Figure 20B), and the cumulative release of tannic acid from the corresponding membranes was monitored over a period of 22 days. As shown, the cumulative release (or release rate) of tannic acid from the sintered membranes was approximately half of the cumulative release (or release rate) of tannic acid from the unsintered membranes over the same period.

[0157] Although several embodiments of the present invention have been described and illustrated herein, those skilled in the art will readily devise various other devices and / or structures for performing the functions described herein and / or achieving the results and / or one or more of the advantages, and each of such variations and / or modifications is considered to be within the scope of the present invention. More generally, those skilled in the art will readily recognize that all parameters, dimensions, materials, and configurations described herein are meant to be exemplary, and that actual parameters, dimensions, materials, and / or configurations will depend on the specific application or applications for which the teachings of the present invention are used. Those skilled in the art will recognize or be able to ascertain using only routine experimentation many equivalents to the specific embodiments of the present invention described herein. Therefore, it should be understood that the foregoing embodiments are presented by way of example only, and that, within the scope of the appended claims and their equivalents, the present invention may be practiced otherwise than as specifically described and claimed. The present invention is directed to each individual feature, system, article, material, kit, and / or method described herein. In addition, any combination of two or more such features, systems, articles, materials, kits, and / or methods, provided such features, systems, articles, materials, kits, and / or methods are not mutually inconsistent, is included within the scope of the present invention.

Claims

1. A macroencapsulation device comprising: first film layer; a second film layer disposed on the first film layer, wherein the first film layer and the second film layer are bonded together to form a seal that extends at least partially around an interior volume disposed between the first film layer and the second film layer, wherein the first film layer and / or the second film layer include pores; and At least one immunomodulatory compound, the at least one immunomodulatory compound being associated with the first membrane layer and / or the second membrane layer, wherein the at least one immunomodulatory compound comprises at least one selected from the group consisting of a toll-like receptor 4 (TLR4) inhibitor, a nuclear factor kappa light-chain enhancer of activated B cells (NF-κB) inhibitor, a colony-stimulating factor 1 receptor (CSF1R) inhibitor, and a NOD, LRR, and pyrin domain-containing protein 3 (NLRP3) inflammasome inhibitor.

2. The macroencapsulated device of claim 1, wherein the at least one immunomodulatory compound is included in a matrix.

3. The macroencapsulated device according to any one of the preceding claims, wherein the at least one immunomodulatory compound is disposed on the first membrane layer and / or the second membrane layer.

4. The macroencapsulated device according to any one of the preceding claims, wherein the at least one immunomodulatory compound is disposed on a frame of the macroencapsulated device.

5. The macroencapsulated device according to any one of the preceding claims, wherein the at least one immunomodulatory compound is disposed in the pores of the first membrane layer and / or the second membrane layer.

6. The macroencapsulated device according to any one of the preceding claims, wherein the at least one immunomodulatory compound is fixed to the first membrane layer and / or the second membrane layer via a linker.

7. The macroencapsulated device of any one of the preceding claims, wherein when implanted, the macroencapsulated device exhibits a reduction in a pro-inflammatory response compared to a macroencapsulated device without the at least one immunomodulatory compound under otherwise substantially identical conditions.

8. The macroencapsulated device of any one of the preceding claims, wherein the first and second film layers are bonded along the perimeters of the first and second films to form the interior volume therebetween.

9. The macroencapsulated device according to any one of the preceding claims, wherein the first membrane layer and / or the second membrane layer is semi-permeable.

10. The macroencapsulated device of any preceding claim, further comprising a frame extending along at least a portion of the perimeter of the first and second membrane layers.

11. The macroencapsulated device of any one of the preceding claims, wherein the first and second membrane layers are configured to prevent passage of a cell population from the device.

12. The macroencapsulated device of claim 11, further comprising the cell population disposed in the interior volume.

13. The macroencapsulated device according to any one of claims 11 to 12, wherein the cell population comprises at least one selected from the group consisting of pancreatic progenitor cells, endocrine cells, alpha cells, delta cells, and beta cells.

14. The microencapsulated device of any one of claims 11 to 12, wherein the cell population comprises NKX6.1-positive, ISL1-positive cells.

15. A method for delivering a therapeutic composition produced by a cell population, the method comprising: allowing the therapeutic composition to diffuse across at least one membrane layer that at least partially encapsulates the cell population; and The tissue surrounding the at least one membrane layer is exposed to at least one immunomodulatory compound selected from the group consisting of a toll-like receptor 4 (TLR4) inhibitor, a nuclear factor kappa light-chain enhancer of activated B cells (NF-κB) inhibitor, a colony-stimulating factor 1 receptor (CSF1R) inhibitor, and a NOD, LRR, and pyrin domain-containing protein 3 (NLRP3) inflammasome inhibitor.

16. The method of claim 15, wherein the at least one immunomodulatory compound is included in a matrix.

17. The method of claim 16, wherein the matrix comprises a hydrogel.

18. The method of any one of claims 15 to 17, wherein exposing the tissue surrounding the at least one membrane layer to the at least one immunomodulatory compound comprises releasing the at least one immunomodulatory compound from the at least one membrane layer.

19. The method of any one of claims 15 to 18, wherein exposing the tissue surrounding the at least one membrane layer to the at least one immunomodulatory compound comprises applying the at least one immunomodulatory compound to the tissue surrounding the at least one membrane layer.

20. The method of any one of claims 15 to 19, wherein the therapeutic composition is released across the at least one membrane layer to tissue within the subject over a period of at least 2 weeks.

21. The method of any one of claims 15 to 20, wherein the at least one immunomodulatory compound is exposed to the tissue over a period of at least 1 day and at most 2 weeks.

22. The method of any one of claims 15 to 21, wherein the therapeutic composition comprises a therapeutic agent selected from the group consisting of insulin and glucagon.

23. The method according to any one of claims 15 to 22, wherein the cell population comprises at least one selected from the group consisting of pancreatic progenitor cells, endocrine cells, alpha cells, delta cells, and beta cells.

24. The method of any one of claims 15 to 22, wherein the cell population comprises NKX6.1-positive, ISL1-positive cells.

25. The method of any one of claims 15 to 24, further comprising increasing the amount of anti-inflammatory cytokines produced relative to pro-inflammatory cytokines when the tissue is exposed to the immunomodulatory compound.

26. An immunomodulatory material comprising: matrix; and Included in the matrix is ​​at least one immunomodulatory compound selected from the group consisting of a toll-like receptor 4 (TLR4) inhibitor, a nuclear factor kappa light-chain enhancer of activated B cells (NF-κB) inhibitor, a colony-stimulating factor 1 receptor (CSF1R) inhibitor, and a NOD, LRR, and pyrin domain-containing protein 3 (NLRP3) inflammasome inhibitor.

27. The immunomodulatory material of claim 26, wherein the matrix comprises a hydrogel.

28. The immunomodulatory material according to any one of claims 26 to 27, wherein the immunomodulatory material comprises a plurality of microparticles and / or nanoparticles.

29. The immunomodulatory material according to any one of claims 26 to 28, wherein the matrix comprises a protein gel.

30. The immunomodulatory material according to any one of claims 26 to 29, wherein the matrix is ​​selected from the following group: fibrin glue, alginate, collagen gel, polyethylene glycol (PEG), polylactic-co-glycolic acid (PLGA), polycaprolactone (PCL), matrigel, poly-L-lactide (PLLA) and / or polyglycolic acid (PGA).

31. The immunomodulatory material according to any one of claims 26 to 30, wherein the matrix comprises fibrin glue comprising fibrinogen at a concentration between 10 wt% and 100 wt%.

32. The immunomodulatory material according to any one of claims 26 to 31 , wherein the immunomodulatory material comprises a cell population.

33. The immunomodulatory material according to claim 32, wherein the cell population comprises at least one cell selected from the group consisting of pancreatic progenitor cells, endocrine cells, α cells, δ cells, and β cells.

34. The immunomodulatory material according to any one of claims 32 to 33, wherein the cell population comprises NKX6.1-positive, ISL1-positive cells.

35. A composition comprising: cell populations; and At least one immunomodulatory compound selected from the group consisting of a toll-like receptor 4 (TLR4) inhibitor, a nuclear factor kappa light-chain enhancer of activated B cells (NF-κB) inhibitor, a colony-stimulating factor 1 receptor (CSF1R) inhibitor, and a NOD, LRR, and pyrin domain-containing protein 3 (NLRP3) inflammasome inhibitor.

36. The composition of claim 35, wherein the composition comprises a matrix.

37. The composition of any one of claims 35 to 36, wherein the cell colony is contained in a matrix.

38. The composition of claim 36 or 37, wherein the matrix comprises a hydrogel.

39. The composition of any one of claims 35 to 38, wherein the immunomodulatory compound is contained in a matrix.

40. The composition of any one of claims 36 to 39, wherein the matrix comprises a protein gel.

41. The composition of any one of claims 36 to 40, wherein the matrix is ​​selected from the group consisting of fibrin glue, alginate, collagen gel, polyethylene glycol (PEG), polylactic-co-glycolic acid (PLGA), polycaprolactone (PCL), matrigel, poly-L-lactide (PLLA) and / or polyglycolic acid (PGA).

42. The composition of any one of claims 36 to 41, wherein the matrix comprises fibrin glue comprising fibrinogen at a concentration of between 10 wt% and 100 wt%.

43. The composition of any one of claims 35 to 42, wherein the cell population comprises at least one cell selected from the group consisting of pancreatic progenitor cells, endocrine cells, alpha cells, delta cells, and beta cells.

44. The composition of any one of claims 35 to 43, wherein the cell population comprises NKX6.1-positive, ISL1-positive cells.

45. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the at least one immunomodulatory compound comprises the TLR4 inhibitor.

46. ​​The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the at least one immunomodulatory compound comprises the NF-κB inhibitor.

47. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the at least one immunomodulatory compound comprises the CSF1R inhibitor.

48. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the at least one immunomodulatory compound comprises the NLRP3 inflammasome inhibitor.

49. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the CSF1R inhibitor comprises pexidartinib.

50. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the TLR4 comprises at least one of TAK242 and tannic acid or a derivative or analog thereof.

51. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the NF-kB inhibitor comprises at least one of TAK242 and tannic acid or a derivative or analog thereof.

52. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the NLRP3 inflammasome inhibitor comprises MCC950 or a derivative or analog thereof.

53. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the TLR4 and / or NF-kB inhibitor comprises a polyphenol or a derivative or analog thereof.

54. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the at least one immunomodulatory compound is present at a concentration greater than or equal to 0.005 mmol / L and less than or equal to 12 mmol / L.

55. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the at least one immunomodulatory compound comprises MCC950 at a concentration greater than or equal to 0.04 mmol / L and less than or equal to 4 mmol / L.

56. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the at least one immunomodulatory compound comprises pesidatinib at a concentration greater than or equal to 0.01 mmol / L and less than or equal to 0.8 mmol / L.

57. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the at least one immunomodulatory compound comprises TAK242 at a concentration greater than or equal to 0.3 mmol / L and less than or equal to 12 mmol / L.

58. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the at least one immunomodulatory compound comprises tannic acid at a concentration greater than or equal to 0.02 mmol / L and less than or equal to 2 mmol / L.

59. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the at least one immunomodulatory compound is present in a therapeutic dose greater than or equal to 0.1 μM and less than or equal to 4 μM.

60. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the at least one immunomodulatory compound comprises MCC950 at a therapeutic dose of between or equal to 0.25 μM and less than or equal to 4 μM.

61. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the at least one immunomodulatory compound comprises a therapeutic dose of between or equal to 0.125 μM and less than or equal to 0.25 μM of pesidatinib.

62. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the at least one immunomodulatory compound comprises TAK242 at a therapeutic dose of between or equal to 2 μM and less than or equal to 4 μM.

63. The macroencapsulated device, method, composition or immunomodulatory material of any preceding claim, wherein the at least one immunomodulatory compound comprises a therapeutic dose of tannic acid between or equal to 0.25 μM and less than or equal to 0.5 μM.

64. A method of treating a patient with type 1 diabetes, the method comprising administering to the patient in need thereof a composition according to any one of claims 35 to 63.

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