Covalent photo-crosslinked polysaccharides and methods of use thereof

Abstract

Described herein are polysaccharide polymers capable of covalently cross-linking with another portion, such as another polysaccharide polymer, as well as related compositions, hydrogel capsules comprising the polysaccharide polymers, and uses thereof.

Description

[0001] Priority Declaration

[0002] This application claims priority to U.S. Application No. 63 / 357,874, filed on July 1, 2022, and U.S. Application No. 63 / 452,125, filed on March 14, 2023. The disclosure of each of the foregoing applications is incorporated herein by reference in its entirety. Background Art

[0003] The function of implanted devices depends largely on the recipient's biological immune response pathways (Anderson et al., Semin. Immunol. 20:86–100 (2008); Langer, Adv. Mater. 21:3235–3236 (2009)). Modulation of the immune response can have beneficial effects on the fidelity and function of these devices. Therefore, there is a need in the art for new compounds, compositions, and devices to achieve this goal. Summary of the Invention

[0004] Described herein are polysaccharide polymers that can be covalently cross-linked with another part, such as another polysaccharide polymer, and related compositions, hydrogel capsules, and their uses comprising the polysaccharide polymers. In one embodiment, the polysaccharide polymer comprises a photoactive cross-linking moiety (such as a compound of formula (IV)) or a pharmaceutically acceptable salt thereof. In one embodiment, the polysaccharide polymer comprises a photoactive cross-linking moiety (e.g., a compound of formula (IV)) and a compound of formula (I) (e.g., a defibrotic compound) or a pharmaceutically acceptable salt thereof. These polysaccharide polymers can be incorporated into hydrogel capsules that can encapsulate cells. Incorporating a photoactive cross-linking agent into the polysaccharide polymer and then into a hydrogel capsule incorporated with the polysaccharide polymer can allow for the regulation of certain properties of the hydrogel capsule, including capsule diameter, stability, and integrity.

[0005] The details of one or more embodiments of the present invention are set forth herein. Other features, objects, and advantages of the invention will be apparent from the detailed description, drawings, examples, and claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0007] Figure 1 Shown are representative LC-UV chromatograms used to characterize the components in the photoactive crosslinker reaction.

[0008] Figure 2A-2C Representative images of two-compartment alginate hydrogel capsules are shown. Figure 2A Figure 2 is an image of an alginate hydrogel capsule in which both the inner and outer compartments contain a blend of VLVG / SLG100 alginate. Figure 2Bis an image of an alginate hydrogel capsule in which both the inner and outer compartments contain a blend of modified and unmodified alginates (VLVG / SLG100), and in which the inner compartment contains dextrin beads to simulate mammalian cells. Figure 2C is an image of a hydrogel in which both the inner and outer compartments contain a blend of methacrylamide-modified VLVG / SLG100 (70 / 30), and in which the inner compartment contains dextrin beads to simulate mammalian cells.

[0009] Figure 3A-3C Show Figure 2A-2C Representative images of alginate hydrogel capsules after one month of storage in buffer.

[0010] Figures 4A-4D Representative images comparing the size and stability of doubly cross-linked alginate hydrogel capsules prepared with different concentrations of photoactive cross-linker: 7% photoactive cross-linker ( Figure 4A ); 5% photoactive crosslinker ( Figure 4B ); 3% photoactive crosslinker ( Figure 4C ); and 1.5% photoactive crosslinker ( Figure 4D ).

[0011] Figure 4E To show Figures 4A-4D Graph of the corresponding burst strengths of the alginate hydrogel capsules described in .

[0012] Figure 5 The average burst strength of exemplary hydrogel capsules (2) comprising a covalently cross-linked polymer comprising Compound 101 as described in Example 7 was compared to that of ionically cross-linked alginate hydrogels (2).

[0013] Figure 6 The average burst strength of exemplary hydrogel capsules (2) comprising a covalently cross-linked polymer comprising Compound 111 as described in Example 7 was compared to that of ionically cross-linked alginate hydrogels (2).

[0014] Figure 7A-7B The hydrogel capsule 1 ( Figure 7A ) and hydrogel capsule 2 ( Figure 7B ) is an exemplary microscopic image of .

[0015] Figure 8A-8B Shown are exemplary microscopic images of doubly cross-linked alginate hydrogel capsules containing exemplary mammalian cells.

[0016] Figure 9Bar graph comparing the fold change in macrophage adhesion of various modified alginate hydrogel capsules described herein; (1): empty spheres; (2)-(5): doubly cross-linked polymer hydrogel capsules containing exemplary mammalian cells.

[0017] Figures 10A-10B The viability percentage of exemplary mammalian cells encapsulated in double cross-linked alginate hydrogel capsules is shown ( Figure 10A ) and total cell number ( Figure 10B ); (1) and (3) were performed at room temperature; (2) and (4) were performed at 4°C.

[0018] Figures 11A-11B UV exposure time is shown ( Figure 11A ) and UV intensity ( Figure 11B ) on the bursting strength of exemplary modified alginate hydrogel capsules.

[0019] Figures 12A-12B UV exposure time is shown ( Figure 12A ) and UV intensity ( Figure 12B ) on the viability of mammalian cells encapsulated in exemplary modified alginate hydrogel capsules.

[0020] Figure 13 is a graph showing the effect of UV exposure on the rate of release of encapsulated dextran polymers from exemplary modified alginate polymers described herein.

[0021] Figure 14 Graphs comparing the viability of mammalian cells encapsulated in different exemplary covalently cross-linked modified alginate capsules described herein over time; (1) ionically cross-linked alginate hydrogel capsules; (2) ionically cross-linked alginate hydrogel capsules comprising RGD peptide; (3) and (5) doubly cross-linked alginate hydrogel capsules; (4) doubly cross-linked alginate hydrogel capsules comprising RGD peptide.

[0022] Figure 15 Shown are the effects of different concentrations of modified alginate polymers on the cytotoxicity of ARPE-19 cells.

[0023] Figures 16A-16C The in vivo retention time is shown to be dependent on compound retention for exemplary modified alginate hydrogel capsules described herein ( Figure 16A ), structural integrity ( Figure 16B ) and mechanical strength ( Figure 16C ); light grey represents doubly cross-linked alginate hydrogel capsules, and dark grey represents ionically cross-linked alginate hydrogel capsules.

[0024] Figure 17Shown are the effects of in vivo residence time on the viability of exemplary mammalian cells encapsulated in modified alginate hydrogel capsules described herein; light grey represents doubly cross-linked hydrogel capsules, and dark grey represents ionically cross-linked alginate hydrogel capsules.

[0025] Figures 18A-18B The effect of different mole percentages of (i) the compound of formula (I) and (ii) the compound of formula (IV) on the size ( Figure 18A ), initial rupture strength ( Figure 18B ) and absolute burst strength ( Figure 18C ); diamonds represent 1X compound 101; circles represent 2X compound 101; and squares represent 3X compound 101.

[0026] Figure 19 The viability of exemplary mammalian cells in the double-crosslinked alginate hydrogel capsules of Example 4 is shown.

[0027] Figure 20 Graphs showing relative PFO responses on: (1) empty spheres; (2) doubly cross-linked alginate hydrogel capsules containing exemplary cells as described herein; and (3) polystyrene beads.

[0028] Figures 21A-21C Varying the batch size has an impact on the size of the two-compartment hydrogel capsules described herein ( Figure 21A ),strength( Figure 21B ) and IgG absorption ( Figure 21C The hydrogel capsules comprised the same composition as described in Example 9 (Hydrogel Capsule #1).

[0029] Figures 22A-22B Effect of varying the density of the compound of formula (IV) on insulin diffusion from the two-compartment hydrogel capsules described herein ( Figure 22A ) and IgG absorption ( Figure 22B ) were compared.

[0030] Figures 23A-23C The effect of varying the density of the compound of formula (IV) on the strength of the two-compartment hydrogel capsules described herein is shown. Figure 23A ), flexibility ( Figure 23B ) and size ( Figure 23C ), where (1; circle) refers to the control hydrogel capsule, and (2; square), (3; triangle), and (4; diamond) refer to hydrogel capsules containing 75%, 62%, and 50% reduction of Compound 205, respectively. The hydrogel capsules contained the same composition as described in Example 9 (Hydrogel Capsule #1).

[0031] Figures 24A-24D The effect of varying the density of the compound of formula (IV) on the strength of the two-compartment hydrogel capsules described herein further comprising exemplary mammalian cells is shown ( Figure 24A ), flexibility ( Figure 24B ),size( Figure 24C ) and IgG absorption ( Figure 24D ), where (1; circles) refers to control hydrogel capsules, and (2; squares) and (3; triangles) refer to hydrogel capsules containing 62% and 50% reduction of compound 205, respectively. Sample (4) refers to ionically cross-linked spheres in the absence of a covalent cross-linker; sample (5) refers to doubly cross-linked spheres, wherein the inner and outer compartments comprise alginate modified with compounds 101 and 205 (hydrogel capsule #2 from Example 9); and (6) refers to doubly cross-linked spheres further comprising exemplary mammalian cells, wherein the inner and outer compartments comprise alginate modified with compounds 101 and 205 (hydrogel capsule #2 from Example 9).

[0032] Figures 25A-25D The effect of varying UV irradiation conditions on the size of the two-compartment hydrogel capsules described herein ( Figure 25A ),strength( Figure 25B ), flexibility ( Figure 25C ) and IgG absorption ( Figure 25D ). The hydrogel capsules tested were the hydrogel capsules described in Example 9 (hydrogel capsule #1). Sample (1) refers to ionically crosslinked spheres in the absence of a covalent crosslinker; sample (2) refers to doubly crosslinked spheres, wherein the inner and outer compartments comprise alginate modified with compounds 101 and 205 (hydrogel capsule #2 from Example 9); sample (3) refers to doubly crosslinked spheres, wherein the inner and outer compartments comprise alginate modified with compounds 101 and 205 (hydrogel capsule #2 from Example 9), further comprising exemplary mammalian cells; and sample (4) refers to an empty well control.

[0033] Figure 26 The volume swelling ratios of exemplary two-compartment hydrogel capsules described herein in different media are shown. The hydrogel capsule tested was the hydrogel capsule described in Example 9 (Hydrogel Capsule #1).

[0034] FIG. 27 shows the effect of different alginate types in the inner compartment on the strength of exemplary two-compartment hydrogel capsules described herein ( Figure 27A ), flexibility ( Figure 27B ), and encapsulated cell viability ( Figure 27C ) and IgG absorption ( Figure 27D). The hydrogel capsules tested were those described in Example 9 (hydrogel capsule #1), but with altered inner compartment alginate. Sample (1) refers to ionically crosslinked spheres in the absence of a covalent crosslinker; sample (2) refers to doubly crosslinked spheres in which the inner and outer compartments comprised alginate modified with compounds 101 and 205 (hydrogel capsule #2 from Example 9); sample (3) refers to doubly crosslinked spheres further comprising exemplary pancreatic islet cells in which the inner and outer compartments comprised alginate modified with compounds 101 and 205 (hydrogel capsule #2 from Example 9); and sample (4) refers to an empty well control.

[0035] Figure 28 Graph showing the rate of barium release from exemplary alginate hydrogel capsules. Sample (1) refers to ionically crosslinked spheres in the absence of a covalent crosslinker washed 15 times with buffer; sample (2) refers to hydrogel capsule #1 from Example 9 washed 8 times with buffer; and sample (3) refers to hydrogel capsule #1 from Example 9 washed with EDTA.

[0036] Figure 29 Bar graph showing residual barium in exemplary alginate hydrogel capsules (Hydrogel Capsule #1 from Example 9) after an increasing number of washes with CMRL or HPLM medium.

[0037] Figure 30 is a schematic diagram showing an overview of the indirect curing process.

[0038] Figures 31A-31C The indirect covalent cross-linking method is shown to improve the strength of the exemplary two-compartment hydrogel capsules described herein ( Figure 31A ),size( Figure 31B ), insulin diffusion ( Figure 31C )’s impact.

[0039] Figure 32 is a schematic diagram depicting exemplary architectures of the polymers and associated hydrogel capsules described herein. DETAILED DESCRIPTION

[0040] The present disclosure provides a polysaccharide polymer comprising a photoactive cross-linking portion and a compound of formula (I), as well as related compositions and hydrogel capsules comprising the polysaccharide polymer, and preparation methods and uses thereof.

[0041] Abbreviations and definitions

[0042] In order to make the present disclosure more easily understood, certain technical and scientific terms are specifically defined below. Unless specifically defined elsewhere in this document, all other technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this invention belongs.

[0043] As used herein, including the appended claims, singular forms such as "a," "an," and "the" include their corresponding plural referents unless the context clearly dictates otherwise.

[0044] "About" or "approximately" when used herein means to modify a numerically defined parameter (e.g., a physical description of a hydrogel capsule such as diameter, sphericity, the number of cells encapsulated therein, the number of capsules in a formulation) to mean that the recited value is within the acceptable functional range of the defined parameter as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, such as the limitations of the measurement system, including the acceptable error range of the measurement system. For example, "about" can mean a range of 20% above and below the recited value. As a non-limiting example, a hydrogel capsule defined as having a diameter of approximately 1.5 millimeters (mm) and encapsulating approximately 5 million (M) cells can have a diameter of 1.2 to 1.8 mm and can encapsulate 4 M to 6 M cells. As another non-limiting example, a formulation of approximately 100 devices (e.g., hydrogel capsules) includes a formulation having 80 to 120 devices. In some embodiments, the term "about" means that the modified parameter can vary by as much as 15%, 10%, or 5% above and below the stated value of the parameter. Alternatively, particularly with respect to certain properties of the devices described herein, such as hydrogel capsule integrity of the defibrotizing compound, the term "about" can mean within an order of magnitude above and below the recited value, e.g., within 5-fold, 4-fold, 3-fold, 2-fold, or 1-fold.

[0045] As used herein, the term "acquire" or "acquiring" refers to the process of obtaining a value (e.g., a numerical value) or an image or physical entity (e.g., a sample) by "directly obtaining" or "indirectly obtaining" a value or physical entity. "Directly obtaining" means performing a process (e.g., performing an analytical method or protocol) to obtain a value or physical entity. "Indirectly obtaining" refers to receiving the value or physical entity from another party or source (e.g., a third-party laboratory that directly obtains a physical entity or value). Directly obtaining a value or physical entity includes performing a process that includes a physical change of a physical substance or the use of a machine or device. Examples of directly obtaining a value include obtaining a sample from a human subject. Directly obtaining a value includes performing a process that uses a machine or device, such as using a fluorescence microscope to obtain fluorescence microscopy data.

[0046] As used herein, "Administer," "administering," or "administration" refers to implanting, absorbing, ingesting, injecting, or otherwise introducing an entity described herein (e.g., a hydrogel capsule, device, or a formulation of a hydrogel capsule or device) into a subject, or providing such an entity to a subject for administration.

[0047] As used herein, "defibrotic" means a compound or material that reduces the foreign body reaction (FBR). For example, the amount of FBR induced in a biological tissue by implanting a device (e.g., a hydrogel capsule) comprising a defibrotic compound (e.g., a hydrogel capsule comprising a polymer covalently modified with a compound listed in Table 3) into the tissue is lower than the FBR induced by implanting an afibrotic-null reference device, i.e., a device lacking any defibrotic compound but having substantially the same composition (e.g., the same CBP-polymer, one or more of the same cell types) and structure (e.g., size, shape, number of compartments). In one embodiment, the FBR is induced using an assay known in the art, such as described in WO 2017 / 075630 or using Vegas, A. et al., Nature Biotechnol One or more of the assays / methods described (above) (e.g., subcutaneous tissue protease measurement of implanted capsules, Masson's trichrome staining (MT), hematoxylin or eosin staining of tissue sections, quantification of collagen density, cell staining and confocal microscopy of macrophages (CD68 or F4 / 80), myofibroblasts (α-muscle actin, SMA), or general cell deposition, quantification of 79 RNA sequences for known inflammatory factors and immune cell markers, or FACS analysis of macrophages and neutrophils on the device (e.g., capsule) retrieved after 14 days in the intraperitoneal space of a suitable test subject (e.g., immunocompetent mouse)) assesses the extent of FBR through an immune response in tissue containing the implanted device (e.g., hydrogel capsule), which may include, for example, protein adsorption, macrophages, multinucleated foreign body giant cells, fibroblasts, and angiogenesis. In one embodiment, FBR is assessed by measuring the level of one or more immune response biomarkers (e.g., cathepsins, TNF-α, IL-13, IL-6, G-CSF, GM-CSF, IL-4, CCL2, or CCL4) in tissue containing the implant. In some embodiments, the FBR induced by a device of the invention (e.g., a hydrogel capsule comprising a defibrotic compound disposed on its outer surface) is at least about 80%, about 85%, about 90%, about 95%, about 99%, or about 100% lower than the FBR induced by a reference device without FBR, e.g., a device that is substantially identical to the device being tested or claimed except for the lack of a device for reducing FBR (e.g., a hydrogel capsule that does not comprise a defibrotic compound but is otherwise substantially identical to the claimed capsule). In some embodiments, the FBR (e.g., the level of one or more biomarkers) is measured after about 30 minutes, about 1 hour, about 6 hours, about 12 hours, about 1 day, about 2 days, about 3 days, about 4 days, about 1 week, about 2 weeks, about 1 month, about 2 months, about 3 months, about 6 months, or more.

[0048] As used herein, "cell" refers to an engineered cell or an unengineered cell. In one embodiment, the cell is an immortalized cell or an engineered cell derived from an immortalized cell. In one embodiment, the cell is a living cell, e.g., viable as measured by any technique described herein or known in the art.

[0049] As used herein, "cell binding peptide (CBP)" means a linear or cyclic peptide comprising an amino acid sequence of a cell binding domain of a ligand derived from a cell adhesion molecule (CAM) (e.g., a cell adhesion molecule that mediates cell-matrix attachment or cell-cell attachment). The length of the CBP is less than 50, 40, 30, 25, 20, 15, or 10 amino acids. In one embodiment, the length of the CBP is between 3 and 12 amino acids, between 4 and 10 amino acids, or 3, 4, 5, 6, 7, 8, 9, or 10 amino acids in length. The CBP amino acid sequence may be identical to a naturally occurring binding domain sequence, or may be a conservatively substituted variant thereof. In one embodiment, the CAM ligand is a mammalian protein. In one embodiment, the CAM ligand is a human protein selected from the group of proteins listed in Table 1 below. In one embodiment, the CBP comprises a cell binding sequence listed in Table 1 below, or a conservatively substituted variant thereof. In one embodiment, the CBP comprises at least one of the cell binding sequences listed in Table 1 below. In one embodiment, the CBP consists essentially of the cell binding sequences listed in Table 1 below. In one embodiment, the CBP is an RGD peptide, meaning that the peptide comprises the amino acid sequence RGD (SEQ ID NO: 43) and optionally comprises one or more additional amino acids located at one or both of the N-terminus and / or the C-terminus. In one embodiment, the CBP is a cyclic peptide comprising RGD (SEQ ID NO: 43), such as one of the cyclic RGD peptides described in Vilaca, H. et al., Tetrahedron 70(35): 5420-5427 (2014). In one embodiment, the CBP is a linear peptide comprising RGD (SEQ ID NO: 43) and is less than 6 amino acids in length. In one embodiment, the CBP is a linear peptide consisting essentially of RGD (SEQ ID NO: 43) or RGDSP (SEQ ID NO: 59).

[0050] Table 1: Exemplary CAM ligand proteins and cell binding sequences

[0051]

[0052] As used herein, "CBP-polymer" means a polymer comprising at least one cell-binding peptide molecule covalently attached to a polymer via a linker. In one embodiment, the polymer is not a peptide or polypeptide. In one embodiment, the polymer in the CBP-polymer does not contain any amino acids. In one embodiment, the polymer in the CBP-polymer is a synthetic or naturally occurring polysaccharide, such as an alginate, such as sodium alginate. In one embodiment, the linker is an amino acid linker (i.e., consisting essentially of a single amino acid, or a peptide of several identical or different amino acids) that is linked to the N-terminus or C-terminus of the CBP via a peptide bond. In one embodiment, the C-terminus of the amino acid linker is linked to the N-terminus of the CBP, and the N-terminus of the amino acid linker is linked to at least one side carboxyl group in the polysaccharide via an amide bond. In one embodiment, the structure of the linker-CBP is represented as G (1-4) In one embodiment, the monosaccharide moiety of the CBP-polysaccharide (e.g., CBP-alginate) is not modified by CBP, e.g., the unmodified moiety has a free carboxyl group or lacks a modifiable side carboxyl group. In one embodiment, the number of polysaccharide moieties with covalently attached CBP is less than any one of the following values: 99%, 95%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, 5%, 1%.

[0053] In one embodiment, the density of CBP modifications in a CBP-polymer is estimated by combustion analysis of percent nitrogen. In one embodiment, the CBP-polymer is an RGD-polymer (e.g., RGD-alginate), which is a polymer (e.g., alginate) covalently modified with a linker-RGD molecule (e.g., a peptide consisting essentially of GRGD (SEQ ID NO: 62) or GRGDSP (SEQ ID NO: 60)), and the density of linker-RGD molecule modifications (e.g., conjugation density) is from about 0.05% nitrogen (N) to 1.00% N, from about 0.10% N to about 0.75% N, from about 0.20% N to about 0.50% N, or from about 0.30% N to about 0.40% N, as determined using an assay described herein. In one embodiment, the conjugation density of the linker-RGD modification in RGD-alginate (e.g., MMW alginate covalently modified with GRGDSP (SEQ ID NO: 60)) is 0.1 to 1.0, 0.2 to 0.8, 0.3 to 0.7, 0.3 to 0.6, 0.4 to 0.6 micromoles of linker-RGD moiety per gram of RGD-polymer in solution (e.g., saline solution) with a viscosity of 80 to 120 cP, as determined by any assay capable of quantifying the amount of peptide conjugated to the polymer, e.g., the quantitative peptide conjugation assay described herein. Unless otherwise expressly stated or apparent from the context, a particular recited numerical concentration, concentration range, density, or density range of CBP in a CBP-polymer refers to the concentration or density of conjugated CBP molecules, i.e., it does not include any residual free (e.g., unconjugated) CBP that may be present in the CBP-polymer.

[0054] As used herein, "conservatively modified variants" or "conservative substitutions" refer to variants of reference peptides or polypeptides that are identical to a reference molecule except for having one or more conservative amino acid substitutions in the amino acid sequence. In one embodiment, the conservatively modified variant consists of an amino acid sequence that is at least 70%, 80%, 85%, 90%, 95%, 97%, 98% or 99% identical to the reference amino acid sequence. Conservative amino acid substitutions refer to substitutions of amino acids with similar characteristics (e.g., charge, side chain size, hydrophobicity / hydrophilicity, main chain conformation and rigidity, etc.) that have minimal effect on the biological activity of the resulting substituted peptide or polypeptide. Conservative substitution tables of functionally similar amino acids are well known in the art, and exemplary substitutions grouped by functional characteristics are listed in Table 2 below.

[0055] Table 2. Exemplary conservative amino acid substitution groups.

[0056]

[0057] As used herein, "crosslinked" and variations thereof, such as "crosslinking" or "x-linked," refers to at least one chemical bond (e.g., an ionic bond or a covalent bond) between two or more polymers. In some embodiments, when there are two or more chemical bonds, crosslinking refers to a mixture of covalent and ionic bonds. In some embodiments, when there are two or more chemical bonds, crosslinking refers to covalent bonds of different types (e.g., covalent bonds comprising different or orthogonal functional groups). In some embodiments, when there are two or more chemical bonds, crosslinking refers to covalent bonds of the same type (e.g., covalent bonds comprising the same functional group). In some embodiments, when there are two or more chemical bonds, crosslinking refers to ionic bonds of the same type (e.g., covalent bonds comprising the same ion (e.g., Ba)). 2+ ) of ionic bonds).

[0058] As used herein, "derived from" with respect to cells refers to cells obtained from a tissue, cell line, or cell, which are then optionally cultured, passaged, differentiated, induced, etc. to produce derived cells. For example, mesenchymal stem cells can be derived from mesenchymal tissue and then differentiated into a variety of cell types.

[0059] As used herein, "device" refers to any implantable article described herein (e.g., a particle, a hydrogel capsule, an implant, a medical device). In some embodiments, the device contains cells (e.g., living cells) capable of expressing a therapeutic agent after implantation of the device and has a structure that supports cell viability by allowing cellular nutrients to enter the device. In some embodiments, the device allows metabolic byproducts and / or therapeutic agents produced by the living cells to be released from the device.

[0060] As used herein, "differential volume" refers to the volume of a compartment within a device described herein, excluding the space occupied by another compartment or compartments. For example, the differential volume of a second compartment (e.g., an outer compartment) in a two-compartment device having an inner compartment and an outer compartment refers to the volume within the second compartment excluding the space occupied by the first compartment (the inner compartment).

[0061] As used herein, " effective amount " refers to the amount of a component of a biological response such as to treat a disease, disorder or illness, a device composition, or a device or device composition, such as comprising a cell (such as an engineered cell) or a plurality of hydrogel capsules of an agent (such as a therapeutic agent) produced by a cell (such as an engineered RPE cell). In some embodiments, the term " effective amount " refers to the amount of a component of a device, such as the cell number in the device, the density of the defibrotic compound placed on the surface of the device and / or in the barrier compartment, the density of the photoactive cross-linking agent in the cell compartment. As will be understood by those skilled in the art, effective amount can vary depending on factors such as the following: desired biological endpoint; pharmacokinetics of therapeutic agent, composition or device (such as hydrogel capsule, particle); the illness treated; mode of administration; and age and health status of the subject. Effective amount covers therapeutic and preventive treatments. For example, in order to alleviate FBR, an effective amount of formula (I) compound can reduce the fibrosis on or near an implanted device or prevent the growth or spread of fibrotic tissue on or near an implanted device.

[0062] As used herein, "endogenous nucleic acid" is a nucleic acid that naturally occurs in the cells of a subject.

[0063] As used herein, an "endogenous polypeptide" is a polypeptide that naturally occurs in the cells of a subject.

[0064] As used herein, "engineered cells" are cells with non-natural changes and are typically comprised of nucleic acid sequences (e.g., DNA or RNA) or polypeptides that do not exist (or exist at different levels) in other similar cells under similar conditions of unengineered (exogenous nucleic acid sequences). In one embodiment, engineered cells comprise exogenous nucleic acids (e.g., vectors or altered chromosomal sequences). In one embodiment, engineered cells comprise exogenous polypeptides. In one embodiment, engineered cells comprise exogenous nucleic acid sequences, such as sequences that are not present in unengineered similar cells, such as DNA or RNA. In one embodiment, exogenous nucleic acid sequences are chromosomal, such as exogenous nucleic acid sequences are exogenous sequences placed in endogenous chromosomal sequences. In one embodiment, exogenous nucleic acid sequences are chromosomal or extrachromosomal, such as non-integrated vectors. In one embodiment, exogenous nucleic acid sequences comprise RNA sequences, such as mRNA. In one embodiment, exogenous nucleic acid sequences comprise chromosomal or extrachromosomal exogenous nucleic acid sequences, and the chromosomal or extrachromosomal exogenous nucleic acid sequences comprise sequences that are expressed as RNA, such as mRNA or regulatory RNA. In one embodiment, the exogenous nucleic acid sequence comprises a chromosome or extrachromosomal nucleic acid sequence, and the chromosome or extrachromosomal nucleic acid sequence comprises a coded polypeptide or is expressed as a sequence of a polypeptide. In one embodiment, the exogenous nucleic acid sequence comprises a first chromosome or extrachromosomal exogenous nucleic acid sequence, and the first chromosome or extrachromosomal exogenous nucleic acid sequence regulates the conformation or expression of the second nucleic acid sequence, wherein the second amino acid sequence can be exogenous or endogenous. For example, the engineered cell can comprise the exogenous nucleic acid for controlling the expression of an endogenous sequence. In one embodiment, the engineered cell comprises a polypeptide existing at a level different from that found in unengineered similar cells or at a distribution. In one embodiment, the engineered cell includes the RPE engineered to produce RNA or polypeptide. For example, the engineered cell can comprise an exogenous nucleic acid sequence, and the exogenous nucleic acid sequence comprises a chromosome or extrachromosomal exogenous nucleic acid sequence, and the chromosome or extrachromosomal exogenous nucleic acid sequence comprises and is expressed as RNA, such as the sequence of mRNA or regulatory RNA. In one embodiment, the engineered cell (for example, RPE cell) comprises an exogenous nucleic acid sequence, and the exogenous nucleic acid sequence comprises a chromosome or extrachromosomal nucleic acid sequence, and the chromosome or extrachromosomal nucleic acid sequence comprises a coded polypeptide or is expressed as a sequence of a polypeptide. In one embodiment, the polypeptide is encoded by a codon-optimized sequence to achieve higher expression of the polypeptide than the naturally occurring coding sequence.Codon-optimized sequences can be obtained by commercially available algorithms, such as GeneOptimizer (ThermoFisher Scientific), OptimumGene™ (GenScript, Piscataway, NJ USA), GeneGPS® (ATUM, Newark, CA USA) or Java CodonAdaptation Tool (JCat, www.jcat.de, Grote, A. et al., Nucleic Acids Research, Vol. 33, Supplement No. 2, pp. W526-W531 (2005) production. In one embodiment, the cell of through engineering approaches (for example, RPE cell) comprises the conformation or the exogenous nucleic acid sequence of expression regulating endogenous sequence. In one embodiment, the cell of through engineering approaches (for example, RPE cell) is obtained by colony of the cell of stably transfected or by culture of through engineering approaches (for example, RPE cell) from monoclonal cell line.

[0065] As used herein, "exogenous nucleic acid" is a nucleic acid that does not naturally occur in the cells of a subject.

[0066] As used herein, "exogenous polypeptide" is a polypeptide that does not naturally occur in a subject's cells (e.g., engineered cells). Reference to an amino acid position in a particular sequence refers to the position of the amino acid in a reference amino acid sequence, e.g., the sequence of a full-length mature (after cleavage of the signal peptide) wild-type protein (unless otherwise indicated), and does not exclude the presence of variations (e.g., deletions, insertions, and / or substitutions) at other positions in the reference amino acid sequence.

[0067] Unless otherwise indicated, otherwise " factor VII protein " or " FVII protein " as used herein refer to the polypeptide comprising the amino acid sequence of naturally occurring factor VII protein or its variant, as determined by art-recognized mensuration, and described polypeptide has FVII biological activity, for example, promotes blood coagulation.Naturally occurring FVII exists with single-chain zymogen, zymogen-like two-chain polypeptide and fully activated two-chain form (FVIIa).In some embodiments, mentioning of FVII includes its single-chain and two-chain form, including zymogen-like and FVIIa.The FVII protein that can be produced by equipment described herein (equipment containing engineered RPE cell) includes wild-type primates (for example, mankind), porcine, canine and murine protein, and the variant of this type of wild-type protein, including fragment, mutant, variant with one or more amino acid replacements and / or disappearances.

[0068] Unless otherwise indicated, " factor VIII protein " or " FVIII protein " as used herein refer to the polypeptide comprising the amino acid sequence of naturally occurring factor VIII polypeptides or its variant, as determined by art-recognized mensuration, and the polypeptide has FVIII biological activity, such as coagulant activity. The FVIII protein that can be expressed by equipment described herein (such as equipment containing engineered RPE cells) includes wild-type primates (such as humans), porcine, canine and murine proteins, and the variant of such wild-type protein, including fragment, mutant, variant with one or more amino acid substitutions and / or deletions, B domain deletion (BDD) variant, single-chain variant, and any one of the aforementioned wild-type or variant and the fusion of half-life extension polypeptide. In one embodiment, cell is engineered to encode a precursor factor VIII polypeptide (for example, with a signal sequence) that has a complete or partial deletion of B domains. In one embodiment, cell is engineered to encode a single-chain factor VIII polypeptide.

[0069] Unless otherwise indicated, " factor IX protein " or " FIX protein " as used herein refer to the polypeptide comprising the amino acid sequence of naturally occurring factor IX protein or its variant, as determined by art-recognized mensuration, and the polypeptide has FIX biological activity, such as coagulant activity. FIX is produced as an inactive zymogen, and the inactive zymogen is converted into active form by the factor XIa excision of activating peptide, thus producing heavy chain and light chain linked together by one or more disulfide bonds. The FIX protein that can be produced by equipment described herein (such as equipment containing engineered RPE cells) includes wild-type primates (such as humans), porcine, canine and murine proteins, and the variant of this type of wild-type protein, including fragment, mutant, variant with one or more amino acid substitutions and / or deletions, and any one of the aforementioned wild-type or variant proteins and the fusion of half-life-extending polypeptides. In one embodiment, cell is engineered to encode full-length wild-type human factor IX polypeptide (such as, with signal sequence) or its functional variant. Unless otherwise indicated, " interleukin -2 protein " or " IL-2 protein " as used herein means a polypeptide comprising the amino acid sequence of a naturally occurring IL-2 protein or a variant thereof, as determined by an assay recognized in the art, the polypeptide having IL-2 biological activity, such as IL-2 receptor signaling in activated Treg cells. The IL-2 protein that can be produced by a device as described herein (such as a device containing engineered RPE cells) includes wild-type primates (such as humans), porcine, canine and murine proteins, and variants of such wild-type proteins.

[0070] As used herein, "islet cells" are intended to include any naturally occurring or synthetically produced or modified cells that are intended to partially or completely reproduce, mimic, or otherwise express some or all of the functions of the islet cells of Langerhans. The term "islet cells" includes glucose-responsive insulin-producing cells derived from stem cells (e.g., from an induced pluripotent stem cell line).

[0071] As used herein, the term "mesenchymal stem cell" or "MSFC" refers to a cell derived from a mesodermal lineage or a cell having at least one characteristic characteristic of a cell of a mesodermal lineage, and wherein the MSFC: i) is not in a terminal state of differentiation; and ii) can be terminally differentiated into one or more cell types. MSFC does not include cells of endoderm origin, such as intestinal cells, or cells of ectoderm origin, such as cells derived from the skin, CNS, or neural cells. In one embodiment, MSFC is multipotent. In one embodiment, MSFC is not fully potent. In one embodiment, MSFC comprises one or more of the following characteristics:

[0072] a) it comprises mesenchymal stem cells (MSCs) or cells derived therefrom, including: cells derived from primary cell cultures of MSCs; cells directly isolated (without long-term culture, e.g., fewer than 5 or 10 passages or cell division cycles since isolation) from naturally occurring MSCs, e.g., from humans or other mammals; cells derived from transformed, potent, immortalized, or long-term (e.g., more than 5 or 10 passages or cell division cycles) MSC cultures; and

[0073] b) It comprises cells obtained from less differentiated cells, such as cells that develop, program, or reprogram (e.g., in vitro) into MSCs, or cells that are substantially similar to one or more of naturally occurring MSCs or cells from primary or long-term MSC cultures, except for any genetic engineering, or cells described in a) above. Examples of less differentiated cells from which MSFCs can be derived include IPS cells, embryonic stem cells, or other totipotent or enriched cells; see, e.g., Chen, YS et al. (2012) Stem Cells Transl Med 1(83-95); Frobel, J et al. (2014) Semin Cell Reports 3(3):414-422; Zou, L et al. (2013) Sci Rep 3:2243.

[0074] As used herein, "parathyroid hormone" or "PTH" means a polypeptide or peptide comprising the amino acid sequence of a naturally occurring parathyroid hormone polypeptide or peptide or a variant thereof, e.g., a polypeptide or peptide having PTH biological activity as determined by an art-recognized assay. PTH polypeptides and peptides that can be expressed by the encapsulated cells described herein include wild-type primate (e.g., human), porcine, canine, and murine proteins, as well as variants of such wild-type proteins. Such PTH polypeptides and peptides can consist essentially of the wild-type human sequence of the preproPTH polypeptide (115 amino acids), the proPTH polypeptide (90 amino acids), the mature 84 amino acid peptide (PTH (1-84)), and biologically active variants thereof, such as the truncated variant peptide PTH (1-34).

[0075] As used herein, "photoactive" refers to a compound or moiety that is capable of forming a reactive species upon activation by light (e.g., light of a specific wavelength). In one embodiment, the photoactive compound is chemically inert under standard laboratory conditions but becomes reactive upon activation by light. A "photoactive crosslinker" is a crosslinker compound that is activated to crosslink upon activation by light.

[0076] As used herein, a "polymer composition" is a composition (e.g., a solution, a mixture) comprising one or more polymers. As a class, "polymers" include homopolymers, heteropolymers, copolymers, block polymers, block copolymers and can be natural and synthetic. Homopolymers contain one type of structural unit or monomer, while copolymers contain more than one type of monomer.

[0077] As used herein, "polypeptide" refers to a polymer comprising amino acid residues linked by peptide bonds and having at least two, and in some embodiments at least 10, 50, 75, 100, 150, or 200 amino acid residues.

[0078] As used herein, "Prevention," "prevent," and "preventing" refer to treatment that includes administering or applying a therapy, such as a composition of a device that encapsulates cells (e.g., as described herein), prior to the onset of a disease, disorder, or condition to preclude physical manifestations of the disease, disorder, or condition. In some embodiments, "Prevention," "prevent," and "preventing" require that signs or symptoms of the disease, disorder, or condition have not yet developed or been observed. In some embodiments, treatment includes prevention, and in other embodiments, it does not include prevention.

[0079] "Alternative therapy" or "alternative protein" is a therapeutic protein or its functional fragment that replaces or enhances the beneficial function of a protein that is reduced, present in insufficient amounts, changes (e.g., mutates) or lacks in a subject suffering from a disease or illness associated with a protein that is reduced, changed or lacks. Examples are certain blood coagulation factors in certain blood coagulation disorders or certain lysosomal enzymes in certain lysosomal storage diseases. In one embodiment, alternative therapy or alternative protein provides the function of an endogenous protein. In one embodiment, alternative therapy or alternative protein has the same amino acid sequence of a naturally occurring variant (e.g., wild-type allele or an allele unrelated to the disease) of the replaced protein. In one embodiment, alternative therapy or alternative protein is different from a naturally occurring variant (e.g., wild-type allele or an allele unrelated to the disease, such as an allele carried by a subject) in an amino acid sequence at no more than about 1%, 2%, 3%, 4%, 5%, 10%, 15% or 20% of the amino acid residues.

[0080] As used herein, "RPE cells" refer to cells having one or more of the following characteristics: a) they comprise retinal pigment epithelial cells (RPE) (e.g., using the ARPE-19 cell line (ATCC ®CRL-2302™) or cells derived therefrom (e.g., cells cultured from the ARPE-19 cell line by stable transfection with an exogenous sequence encoding a therapeutic protein, or such cultured ARPE-19 cells are otherwise engineered to express an exogenous protein or other exogenous substance), cells derived from primary cell cultures of RPE cells, cells isolated directly from naturally occurring RPE cells (e.g., from humans or other mammals) (without long-term culture, e.g., fewer than 5 or 10 generations or rounds of cell division since isolation), cells derived from transformed, immortalized, or long-term (e.g., more than 5 or 10 generations or rounds of cell division) b) cells derived from a less differentiated cell, such as a cell that is developed, programmed, or reprogrammed (e.g., in vitro) into an RPE cell, or a cell that is substantially similar to one or more of a naturally occurring RPE cell or a primary or long-term culture of an RPE cell, except for any genetic engineering (e.g., the cell may be derived from an IPS cell); c) a cell having one or more of the following characteristics: i) it expresses one or more of the biomarkers CRALBP, RPE-65, RLBP, BEST1, or αB-crystallin; ii) it does not express one or more of the biomarkers CRALBP, RPE-65, RLBP, BEST1, or αB-crystallin; iii) it is naturally present in the retina and forms a monolayer above the choroidal vessels in Bruch's membrane; iv) it is responsible for epithelial transport, light absorption, secretion, and immunomodulation in the retina; or v) it is synthetically produced or modified by a naturally occurring cell to have properties similar to those of an immortalized RPE cell line (e.g., the ARPE-19 cell line (ATCC 5015); ® In one embodiment, RPE cell described herein is engineered to express therapeutic agent when being encapsulated in the polysaccharide polymer hydrogel capsule that for example modifies through photoactive cross-linking agent.In other embodiments, RPE cell is without engineered.

[0081] When used in this article to refer to two nucleotide sequences or two amino acid sequences, "sequence identity" or "percentage identity" means that when two sequences are compared and aligned over a comparison window or a specified region to obtain maximum correspondence, the two sequences are identical within the specified region, or have identical nucleotides or amino acids at the nucleotide or amino acid positions at a specified percentage within the specified region. Sequence identity can be determined using standard techniques known in the art, including but not limited to any algorithm described in U.S. Patent Application Publication No. 2017 / 02334455. In one embodiment, the specified percentage of identical nucleotide or amino acid positions is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher.

[0082] As used herein, "spherical" means a device (e.g., a hydrogel capsule or other particle) having a curved surface that forms a sphere (e.g., a perfectly round sphere) or a sphere-like shape, which may have waves and undulations on the surface, for example. Spheres and sphere-like objects can be mathematically defined by rotating a circle, ellipse, or combination about each of three perpendicular axes a, b, and c. For a sphere, the three axes are the same length. In general, a sphere-like shape is an ellipsoid whose semi-major axes are within 10%, 5%, or 2.5% of each other (with respect to their average surface). The diameter of a sphere or sphere-like shape is the average diameter, such as the average of the semi-major axes.

[0083] The term "spheroid" as used herein to refer to a device (e.g., a hydrogel capsule or other particle) means that the device has (i) a perfect or classic oblate spheroid or prolate spheroid shape, or (ii) has a surface that is generally spheroidal, e.g., may have corrugations and undulations and / or may be an ellipsoid with semi-major axes within 100% of each other (with respect to its average surface).

[0084] As used herein, "subject" refers to humans or non-human animals. In one embodiment, the subject is a human (i.e., male or female), such as a person of any age group, a pediatric subject (e.g., infant, child, teenager) or an adult subject (e.g., young person, middle-aged person or elderly person). In one embodiment, the subject is a non-human animal, such as a mammal (e.g., mouse, dog, primate (e.g., cynomolgus monkey or rhesus monkey)). In one embodiment, the subject is a commercially relevant mammal (e.g., cattle, pig, horse, sheep, goat, cat or dog) or a bird (e.g., commercially relevant birds, such as chicken, duck, goose or turkey). In certain embodiments, the animal is a mammal. The animal can be male or female and be in any stage of development. The non-human animal can be a transgenic animal.

[0085] As used herein, "total volume" refers to the volume within one compartment of a multi-compartment device, including the space occupied by the other compartment. For example, the total volume of the second compartment (e.g., the outer compartment) of a two-compartment device refers to the volume within the second compartment, including the space occupied by the first compartment. As used herein, the terms "treatment," "treat," and "treating" refer to alleviating, reversing, relieving one or more of the symptoms, manifestations, or potential causes of a disease, disorder, or illness, delaying its onset, or inhibiting its progression. In one embodiment, treatment includes alleviating, reversing, relieving the symptoms of a disease, disorder, or illness, delaying its onset, or inhibiting its progression. In one embodiment, treatment includes alleviating, reversing, relieving the manifestations of a disease, disorder, or illness, delaying its onset, or inhibiting its progression. In one embodiment, treatment includes alleviating, reversing, relieving, alleviating the potential causes of a disease, disorder, or illness, or delaying its onset. In some embodiments, "treatment" requires that signs or symptoms of the disease, disorder, or illness have appeared or have been observed. In other embodiments, treatment can be administered in the absence of signs or symptoms of the disease or illness, such as in preventive treatment. For example, treatment can be administered to a susceptible individual prior to the onset of symptoms (e.g., taking into account a history of symptoms and / or based on genetic or other susceptibility factors). Treatment can also be continued after symptoms have resolved, e.g., to delay or prevent their recurrence. In some embodiments, treatment includes prevention, and in other embodiments, it does not include prevention.

[0086] Unless otherwise indicated, as used herein, "von Willebrand Factor protein" or "VWF protein" means a polypeptide comprising the amino acid sequence of a naturally occurring VWF polypeptide or a variant thereof, which polypeptide has VWF biological activity, e.g., FVIII binding activity, as determined by an art-recognized assay. VWF proteins that can be produced by the devices described herein (e.g., expressed by engineered cells contained in the devices) include wild-type primate (e.g., human), porcine, canine, and murine proteins, as well as variants of such wild-type proteins. Encapsulated cells can be engineered to encode any of the following VWF polypeptides: a 2813 amino acid precursor VWF, a VWF lacking a 22 amino acid signal peptide and optionally lacking a 741 amino acid prepropeptide, a 2050 amino acid mature VWF protein, and truncated variants thereof.

[0087] "VLVG" or "VLVG alginate" refers to a very low viscosity sodium alginate having an average molecular weight of less than about 75 kDa and comprising greater than 60% guluronate units (ie, having a guluronate to mannuronic acid ratio greater than or equal to 1.5).

[0088] "SG100" or "SG100 alginate" refers to sodium alginate having an average molecular weight of about 150 to 250 kDa and comprising greater than 60% guluronic acid units (ie, having a guluronic acid to mannuronic acid ratio greater than or equal to 1).

[0089] Selected chemical definitions

[0090] The definition of specific functional groups and chemical terms is described in more detail below. Chemical elements are identified according to the periodic table CAS version, Handbook of Chemistry and Physics, 75th edition, inside cover, and specific functional groups are generally defined as described therein. In addition, the general principles of organic chemistry and specific functional moieties and reactivity are described in Thomas Sorrell, Organic Chemistry, University Science Books, Sausalito, 1999; Smith and March, March's Advanced Organic Chemistry, 5th edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; and Carruthers, Some Modern Methods of Organic Synthesis, 3rd edition, Cambridge University Press, Cambridge, 1987.

[0091] Abbreviations used herein have their conventional meanings in the fields of chemistry and biology.The chemical structures and formulae illustrated herein are constructed according to the standard rules of chemical valence known in the chemical arts.

[0092] When a range of values ​​is listed, it is intended to encompass each value and subrange within that range. For example, "C1-C6 alkyl" is intended to encompass C1, C2, C3, C4, C5, C6, C1-C6, C1-C5, C1-C4, C1-C3, C1-C2, C2-C6, C2-C5, C2-C4, C2-C3, C3-C6, C3-C5, C3-C4, C4-C6, C4-C5, and C5-C6 alkyl.

[0093] As used herein, "alkyl" refers to a group having a straight chain or branched saturated hydrocarbon group of 1 to 24 carbon atoms ("C1-C 24In some embodiments, an alkyl group has 1 to 12 carbon atoms ("C1-C 12 alkyl”), 1 to 10 carbon atoms (“C1-C 12 In some embodiments, the alkyl group has 2 to 6 carbon atoms ("C2-C6 alkyl"). Examples of C1-C6 alkyl groups include methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), and n-propyl (C1). ), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), isobutyl (C4), n-pentyl (C5), 3-pentyl (C5), pentyl (C5), neopentyl (C5), 3-methyl-2-butanyl (C5), tert-pentyl (C5), and n-hexyl (C6). Additional examples of alkyl include n-heptyl (C7), n-octyl (C8), and the like. Each example of alkyl can independently be optionally substituted, i.e., unsubstituted ("unsubstituted alkyl") or substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent ("substituted alkyl").

[0094] As used herein, "alkenyl" refers to a straight or branched chain hydrocarbon group having 2 to 24 carbon atoms, one or more carbon-carbon double bonds, and no triple bonds ("C2-C 24 In some embodiments, an alkenyl group has 2 to 10 carbon atoms ("C2-C 10 The carbon-carbon double bond(s) may be internal (as in 2-butenyl) or terminal (as in 1-butenyl). Examples of C2-C4 alkenyl groups include ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), and the like. Examples of C2-C6 alkenyl groups include the aforementioned C 2-4 alkenyl as well as pentenyl (C5), pentadienyl (C5), hexenyl (C6), etc. Each instance of an alkenyl group can independently be optionally substituted, i.e., unsubstituted (an "unsubstituted alkenyl") or substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent (a "substituted alkenyl").

[0095] As used herein, the term "alkynyl" refers to a group having a straight chain or branched hydrocarbon group of 2 to 24 carbon atoms, one or more carbon-carbon triple bonds ("C2-C 24 In some embodiments, an alkynyl group has 2 to 10 carbon atoms ("C2-C 10 The carbon-carbon triple bond(s) may be internal (as in 2-butynyl) or terminal (as in 1-butynyl). Examples of C2-C4 alkynyl groups include ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), and the like. Each example of an alkynyl group may independently be optionally substituted, i.e., unsubstituted (an "unsubstituted alkynyl") or substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent (a "substituted alkynyl").

[0096] As used herein, the term "heteroalkyl" refers to a non-cyclic stable straight or branched chain or a combination thereof comprising at least one carbon atom and at least one heteroatom selected from the group consisting of O, N, P, Si, and S, and wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen heteroatom is optionally quaternized. Heteroatoms O, N, P, S, and Si may be placed in any position of the heteroalkyl group. Exemplary heteroalkyl groups include, but are not limited to, -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2, -S(O)-CH3, -CH2-CH2-S(O)2-CH3, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=N-OCH3, -CH=CH-N(CH3)-CH3, -O-CH3, and -O-CH2-CH3. Up to two or three heteroatoms may be consecutive, such as, for example, -CH2-NH-OCH3 and -CH2-O-Si(CH3)3. When "heteroalkyl" is described, the specific heteroalkyl group is then described, such as -CH2O, -NR C R D It will be understood that the terms heteroalkyl and -CH2O or -NR C R Dare not redundant or mutually exclusive. Instead, specific heteroalkyl groups are described to increase clarity. Therefore, the term "heteroalkyl" should not be interpreted herein to exclude specific heteroalkyl groups such as -CH2O, -NR C R D Each instance of heterocyclyl can independently be optionally substituted, i.e., unsubstituted ("unsubstituted heteroalkyl") or substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent ("substituted heteroalkyl").

[0097] Unless otherwise indicated, the terms "alkylene," "alkenylene," "alkynylene," or "heteroalkylene," alone or as part of another substituent, mean a divalent radical derived from an alkyl, alkenyl, alkynyl, or heteroalkylene, respectively. An alkylene, alkenylene, alkynylene, or heteroalkylene group may be described, for example, as a C1-C6-membered alkylene, a C2-C6-membered alkenylene, a C2-C6-membered alkynylene, or a C1-C6-membered heteroalkylene group, where the term "membered" refers to the non-hydrogen atoms within the group. In the case of heteroalkylene groups, heteroatoms may also occupy one or both of the chain ends (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, etc.). Furthermore, for alkylene and heteroalkylene linking groups, the orientation of the linking group is not implied by the direction in which the chemical formula of the linking group is written. For example, the formula -C(O)2R'- may represent both -C(O)2R'- and -R'C(O)2-.

[0098] As used herein, "aryl" refers to a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 π electrons shared in the ring array) having 6-14 ring carbon atoms and zero heteroatoms ("C6-C 14 In some embodiments, an aryl group has 6 ring carbon atoms ("C6 aryl"; e.g., phenyl). In some embodiments, an aryl group has 10 ring carbon atoms ("C 10 "Aryl"; for example, naphthyl, such as 1-naphthyl and 2-naphthyl). In some embodiments, an aryl group has 14 ring carbon atoms ("C 14 "aryl"; for example, anthracenyl). Aryl can be described as, for example, C6-C 10 In some embodiments, the term "membered" refers to a non-hydrogen ring atom within the moiety. Aryl includes phenyl, naphthyl, indenyl, and tetrahydronaphthyl. Each instance of an aryl group can independently be optionally substituted, i.e., unsubstituted (an "unsubstituted aryl") or substituted (a "substituted aryl") with one or more substituents.

[0099] As used herein, "heteroaryl" refers to a radical of a 5-10 membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 π electrons shared in the ring array) having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-10 membered heteroaryl"). In heteroaryl groups containing one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom when valence permits. Heteroaryl bicyclic ring systems can include one or more heteroatoms in one or both rings. "Heteroaryl" also includes ring systems in which a heteroaryl ring as defined above is fused to one or more aryl groups, wherein the point of attachment is on the aryl or heteroaryl ring, and in such cases, the number of ring members refers to the number of ring members in the fused (aryl / heteroaryl) ring system. Bicyclic heteroaryl groups, where one ring contains no heteroatoms (e.g., indolyl, quinolinyl, carbazolyl, etc.), can be attached at either ring, i.e., the ring bearing the heteroatom (e.g., 2-indolyl) or the ring containing no heteroatoms (e.g., 5-indolyl). Heteroaryl groups can be described, for example, as 6-10 membered heteroaryl groups, where the term "membered" refers to non-hydrogen ring atoms within the moiety.

[0100] In some embodiments, heteroaryl is a 5-10 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-10 membered heteroaryl”). In some embodiments, heteroaryl is a 5-8 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-8 membered heteroaryl”). In some embodiments, heteroaryl is a 5-6 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-6 membered heteroaryl”). In some embodiments, 5-6 membered heteroaryl has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, 5-6 membered heteroaryl has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl group has 1 ring heteroatom selected from nitrogen, oxygen, and sulfur. Each instance of a heteroaryl group can independently be optionally substituted, i.e., unsubstituted (an "unsubstituted heteroaryl") or substituted (a "substituted heteroaryl") with one or more substituents.

[0101] Exemplary 5-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyrrolyl, furanyl, and thiophenyl. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include, but are not limited to, tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyridinyl. Exemplary 6-membered heteroaryl groups containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryl groups containing one heteroatom include, but are not limited to, azaphenyl, oxphenyl, and thienyl. Exemplary 5,6-bicyclic heteroaryls include, but are not limited to, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryls include, but are not limited to, naphthyridinyl, pteridinyl, quinolyl, isoquinolyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. Other exemplary heteroaryls include heme and heme derivatives.

[0102] As used herein, the terms "arylene" and "heteroarylene," by themselves or as part of another substituent, refer to a divalent radical derived from an aryl and heteroaryl group, respectively.

[0103] As used herein, "cycloalkyl" refers to a non-aromatic ring system having 3 to 10 ring carbon atoms ("C3-C 10 In some embodiments, the cycloalkyl group has 3 to 8 ring carbon atoms ("C3-C8 cycloalkyl"), 3 to 6 ring carbon atoms ("C3-C6 cycloalkyl"), or 5 to 10 ring carbon atoms ("C5-C8 cycloalkyl"). 10Cycloalkyl"). Cycloalkyl can be described, for example, as a C4-C7 membered cycloalkyl, where the term "membered" refers to the non-hydrogen ring atoms within the moiety. Exemplary C3-C6 cycloalkyls include, but are not limited to, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), and the like. Exemplary C3-C8 cycloalkyls include but are not limited to Not limited to the aforementioned C3-C6 cycloalkyl and cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), cyclooctyl (C8), cyclooctenyl (C8), cubanyl (C8), bicyclo[1.1.1]pentanyl (C5), bicyclo[2.2.2]octyl (C8), bicyclo[2.1.1]hexanyl (C6), bicyclo[3.1.1]heptanyl (C7), etc. Exemplary C3-C 10 Cycloalkyl includes but is not limited to the aforementioned C3-C8 cycloalkyl and cyclononyl (C9), cyclononenyl (C9), cyclodecyl (C9), 10 ), cyclodecenyl (C 10 ), octahydro-1H-indenyl (C9), decahydronaphthyl (C 10 ), spiro[4.5]decyl (C 10 ) and the like. As illustrated by the foregoing examples, in certain embodiments, a cycloalkyl group is monocyclic (a "monocyclic cycloalkyl") or contains a fused, bridged, or spiro ring system, such as a bicyclic ring system (a "bicyclic cycloalkyl"), and may be saturated or partially unsaturated. "Cycloalkyl" also includes ring systems in which a cycloalkyl ring, as defined above, is fused to one or more aryl groups, wherein the point of attachment is on the cycloalkyl ring, and in such cases, the number of carbons continues to refer to the number of carbons in the cycloalkyl ring system. Each instance of a cycloalkyl group may independently be optionally substituted, i.e., unsubstituted (an "unsubstituted cycloalkyl"), or substituted (a "substituted cycloalkyl") with one or more substituents.

[0104] As used herein, "heterocyclyl" refers to a group of a 3 to 10 membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus and silicon ("3-10 membered heterocyclyl"). In heterocyclyl groups containing one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as long as valence permits. The heterocyclyl group can be monocyclic ("monocyclic heterocyclyl") or a fused, bridged or spirocyclic ring system, such as a bicyclic system ("bicyclic heterocyclyl"), and can be saturated or can be partially unsaturated. A heterocyclyl bicyclic ring system can include one or more heteroatoms in one or both rings. "Heterocyclyl" also includes ring systems in which the heterocyclyl ring as defined above is fused to one or more cycloalkyl groups, wherein the point of attachment is on the cycloalkyl or heterocyclyl ring; or ring systems in which the heterocyclyl ring as defined above is fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring, and in such cases, the number of ring members continues to represent the number of ring members in the heterocyclyl ring system. Heterocyclyl can be described as, for example, 3-7 membered heterocyclyl, where the term "member" refers to the non-hydrogen ring atoms within the moiety, i.e., carbon, nitrogen, oxygen, sulfur, boron, phosphorus, and silicon. Each instance of a heterocyclyl can independently be optionally substituted, i.e., unsubstituted ("unsubstituted heterocyclyl") or substituted ("substituted heterocyclyl") with one or more substituents. In certain embodiments, a heterocyclyl is an unsubstituted 3-10 membered heterocyclyl. In certain embodiments, a heterocyclyl is a substituted 3-10 membered heterocyclyl.

[0105] In some embodiments, a heterocyclyl group is a 5-10 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon (“5-10 membered heterocyclyl”). In some embodiments, a heterocyclyl group is a 5-8 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-8 membered heterocyclyl”). In some embodiments, a heterocyclyl group is a 5-6 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-6 membered heterocyclyl”). In some embodiments, a 5-6 membered heterocyclyl group has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, a 5-6 membered heterocyclyl group has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heterocyclyl has one ring heteroatom selected from nitrogen, oxygen, and sulfur. Exemplary 3-membered heterocyclyls containing one heteroatom include, but are not limited to, aziridinyl, oxiranyl, and thioranyl. Exemplary 4-membered heterocyclyls containing one heteroatom include, but are not limited to, azetidinyl, oxetanyl, and thietanyl. Exemplary 5-membered heterocyclyls containing one heteroatom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrophenylthio, dihydrophenylthio, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclyls containing two heteroatoms include, but are not limited to, dioxolanyl, oxathiolanyl, dithiolanyl, and oxazolidin-2-one. Exemplary 5-membered heterocyclyls containing three heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclic groups containing 1 heteroatom include, but are not limited to, piperidinyl, piperazinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclic groups containing 2 heteroatoms include, but are not limited to, piperazinyl, morpholinyl, dithianyl, and dioxanyl. Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to, triazinanyl or thiomorpholinyl-1,1-dioxide. Exemplary 7-membered heterocyclic groups containing 1 heteroatom include, but are not limited to, azepanyl, oxepanyl, and thiepanyl. Exemplary 8-membered heterocyclic groups containing 1 heteroatom include, but are not limited to, azocanyl, oxepanyl, and thiecanyl. Exemplary 5-membered heterocyclic groups fused to a C6 aryl ring (also referred to herein as 5,6-bicyclic heterocycles) include, but are not limited to, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, benzoxazolinone, and the like. Exemplary 6-membered heterocyclyl groups (also referred to herein as 6,6-bicyclic heterocycles) fused to an aryl ring include, but are not limited to, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like.

[0106] As used herein, "amino" refers to the group -NR 70 R 71, where R 70 and R 71 Each independently represents hydrogen, C1-C8 alkyl, C3-C 10 Cycloalkyl, C4-C 10 Heterocyclic group, C6-C 10 Aryl and C5-C 10 In some embodiments, amino refers to NH2.

[0107] As used herein, "cyano" refers to -CN.

[0108]

[0046] As used herein, "halo" or "halogen," by itself or as part of another substituent, means, unless otherwise stated, a fluorine (F), chlorine (Cl), bromine (Br), or iodine (I) atom.

[0109] As used herein, "hydroxy" refers to the group -OH.

[0110] As defined herein, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl groups are optionally substituted (e.g., "substituted" or "unsubstituted" alkyl, "substituted" or "unsubstituted" alkenyl, "substituted" or "unsubstituted" alkynyl, "substituted" or "unsubstituted" heteroalkyl, "substituted" or "unsubstituted" cycloalkyl, "substituted" or "unsubstituted" heterocyclyl, "substituted" or "unsubstituted" aryl, or "substituted" or "unsubstituted" heteroaryl). In general, the term "substituted," whether preceded by the term "optionally" or not, means that at least one hydrogen present on a group (e.g., a carbon or nitrogen atom) is replaced with a permissible substituent, for example, a substituent that results in a stable compound, for example, a compound that does not undergo spontaneous transformation, such as by rearrangement, cyclization, elimination, or other reactions. Unless otherwise indicated, a "substituted" group has a substituent at one or more substitutable positions of the group, and when substitution occurs at more than one position in any given structure, the substituent at each position is the same or different. The term "substituted" is intended to include substitution with all permissible substituents of an organic compound (e.g., any substituent described herein that results in the formation of a stable compound). The present disclosure contemplates any and all such combinations to achieve stable compounds. For purposes of this disclosure, heteroatoms (such as nitrogen) may have hydrogen substituents and / or any suitable substituents that satisfy the valence of the heteroatoms and produce the formation of a stable portion as described herein.

[0111] Two or more substituents may optionally be linked to form an aryl, heteroaryl, cycloalkyl or heterocyclyl group. It has been found that such so-called ring-forming substituents are typically, but not necessarily, attached to a cyclic base structure. In one embodiment, the ring-forming substituents are attached to adjacent members of the base structure. For example, two ring-forming substituents attached to adjacent members of the cyclic base structure form a fused ring structure. In another embodiment, the ring-forming substituents are attached to a single member of the base structure. For example, two ring-forming substituents are attached to a single member of the cyclic base structure, producing a spirocyclic structure. In another embodiment, the ring-forming substituents are attached to non-adjacent members of the base structure.

[0112] The compounds of formula (I) described herein may contain one or more asymmetric centers and are therefore capable of existing in a variety of isomeric forms (e.g., enantiomers and / or diastereomers). For example, the compounds described herein may be in the form of individual enantiomers, diastereomers, or geometric isomers, or may be in the form of stereoisomer mixtures (including racemic mixtures and mixtures enriched in one or more stereoisomers). Isomers may be separated from the mixture by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers may be prepared by asymmetric synthesis. See, e.g., Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, Tables of Resolving Agents and Optical Resolutions, p. 268 (EL Eliel, ed., Univ. of Notre Dame Press, Notre Dame, IN 1972). The present disclosure further encompasses the compounds described herein as individual isomers substantially free of other isomers, as well as, alternatively, as mixtures of different isomers.

[0113] As used herein, a pure enantiomer compound is substantially free of other enantiomers or stereoisomers of the compound (i.e., in enantiomeric excess). In other words, the "S" form of the compound is substantially free of the "R" form of the compound and is therefore an enantiomeric excess of the "R" form. The term "enantiomerically pure" or "pure enantiomer" means that the compound comprises more than 75% by weight, more than 80% by weight, more than 85% by weight, more than 90% by weight, more than 91% by weight, more than 92% by weight, more than 93% by weight, more than 94% by weight, more than 95% by weight, more than 96% by weight, more than 97% by weight, more than 98% by weight, more than 99% by weight, more than 99.5% by weight, or more than 99.9% by weight of the enantiomer. In certain embodiments, weight is the total weight of all enantiomers or stereoisomers based on the compound.

[0114] The compounds of formula (I) described herein may also contain one or more isotopic substitutions. For example, H may be in any isotopic form, including 1 H. 2 H (D or deuterium) and 3 H (or tritium); C can be in any isotopic form, including 12 C. 13 C and 14 C; O can be in any isotopic form, including 16 O and 18 O etc.

[0115] The term "pharmaceutically acceptable salt" is intended to include salts of the active compounds prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein. When the compounds of formula (I) used to prepare the devices of the present disclosure contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base (either neat or in a suitable inert solvent). Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino or magnesium salts, or similar salts. When the compounds used in the present disclosure contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid or phosphorous acid; and salts derived from organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, and the like. Also included are salts of amino acids such as arginine salts; and salts of organic acids such as glucuronic acid or galacturonic acid (see, for example, Berge et al., Journal of Pharmaceutical Science 66: 1-19 (1977)). Certain specific compounds used in the devices of the present disclosure (e.g., particles, hydrogel capsules) contain both basic and acidic functional groups that allow the compound to be converted into base addition salts or acid addition salts. These salts can be prepared by methods known to those skilled in the art. Other pharmaceutically acceptable carriers known to those skilled in the art are also suitable for use in the present disclosure.

[0116] As used herein, "polysaccharide" refers to a polymer of monosaccharides or disaccharide carbohydrates bound together by glycosidic linkages. Polysaccharides can be linear or branched. Exemplary monosaccharides include glucose, galactose, mannose, allose, altrose, talose, idose, gulose, fructose, ribose, arabinose, lyxose, xylose, rhamnose, glucuronic acid, galacturonic acid, uronic acid, mannuronic acid and guluronic acid. Exemplary polysaccharides include alginate, agar, agarose, carrageenan, hyaluronate, amylopectin, glycogen, gelatin, cellulose, amylose, chitin, chitosan or derivatives or variants thereof, for example, as described in Laurienzo (2010), Mar Drugs 9:2435-65.

[0117] The devices of the present disclosure may contain compounds of formula (I) in the form of prodrugs. Prodrugs are compounds that readily undergo chemical changes under physiological conditions to provide compounds useful in preparing the devices of the present disclosure. Additionally, prodrugs can be converted into useful compounds of formula (I) by chemical or biochemical methods in an ex vivo environment.

[0118] Some of the compounds of formula (I) described herein can exist in unsolvated form and solvated form (including hydrated form). In general, solvated form is equivalent to unsolvated form and is encompassed within the scope of the present disclosure. Some of the compounds of formula (I) described herein can exist in multiple crystals or amorphous forms. In general, all physical forms are equivalent for the purposes encompassed by the present disclosure and are intended to fall within the scope of the present disclosure.

[0119] The term "solvate" refers to a form of a compound that is associated with a solvent, typically by a solvolysis reaction. This physical association may include hydrogen bonding. Commonly used solvents include water, methanol, ethanol, acetic acid, DMSO, THF, diethyl ether, and the like. The compounds described herein can be prepared, for example, in crystalline form and can be solvated. Suitable solvates include pharmaceutically acceptable solvates and further include stoichiometric solvates and non-stoichiometric solvates.

[0120] The term "hydrate" refers to a compound associated with water. Typically, the number of water molecules contained in a hydrate of a compound is in a certain ratio to the number of compound molecules in the hydrate. Thus, a hydrate of a compound can be represented, for example, by the general formula R∙x HO, where R is the compound and x is a number greater than 0.

[0121] As used herein, the term "tautomer" refers to a compound structure that is an interchangeable form and that varies in terms of hydrogen atom and electron displacement. Thus, by the movement of π electrons and atoms (usually H), the two structures can be in equilibrium. For example, enols and ketones are tautomers because they can be rapidly converted to each other by treatment with an acid or base. Tautomeric forms may be relevant to the optimization of chemical reactivity and biological activity of related compounds.

[0122] As used herein, the symbol " ” refers to attachment to a surface of an entity, such as a polymer (e.g., a hydrogel-forming polymer such as alginate) or an implantable device (e.g., a particle, a hydrogel capsule). The attachment represented by " can refer to direct attachment to an entity, such as a polymer (e.g., alginate) or an implantable element, or can refer to linkage to an entity through an attachment group. As described herein, an "attachment group" refers to a moiety used to link a compound of Formula (I)-(IV) to an entity (e.g., a hydrogel capsule or implantable device as described herein), and can comprise any attachment chemistry known in the art. A list of exemplary attachment groups is summarized in Bioconjugate Techniques (3rd ed., Greg T. Hermanson, Waltham, MA: Elsevier, Inc, 2013), which is incorporated herein by reference in its entirety. In some embodiments, the attachment group includes alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -C(O)-, -OC(O)-, -N(R C )-、-N(R C )C(O)-、-C(O)N(R C )-、-N(R C )N(R D )-、-NCN-、-C(=N(R C )(R D ))O-、-S-、-S(O) x -、-OS(O) x -、-N(R C )S(O) x -、-S(O) x N(R C )-、-P(R F ) y -、-Si(OR A )2-、-Si(R G )(OR A )-、-B(OR A )- or metal, where R A 、R C 、R D 、R F 、R G Each of x and y is independently as described herein. In some embodiments, the attachment group comprises an amine, a ketone, an ester, an amide, an alkyl, an alkenyl, an alkynyl, or a thiol. In some embodiments, the attachment group is a cross-linking agent. In some embodiments, the attachment group is -C(O)(C1-C6-alkylene)-, wherein the alkylene is replaced by R 1 Replace, and R 1 As described herein. In some embodiments, the attachment group is -C(O)(C1-C6-alkylene)-, wherein the alkylene is substituted with 1-2 alkyl groups (e.g., 1-2 methyl groups). In some embodiments, the attachment group is -C(O)C(CH3)2 -. In some embodiments, the attachment group is -C(O)(methylene)-, wherein the alkylene group is substituted with 1-2 alkyl groups (e.g., 1-2 methyl groups). In some embodiments, the attachment group is -C(O)CH(CH3)-. In some embodiments, the attachment group is -C(O)C(CH3)-.

[0123] As used herein, the terms "covalent," "covalent bond," and "covalent bonding" refer to a type of chemical bond involving the sharing of electrons between two adjacent atoms. Examples of covalent bonds include bonds formed between carbon and hydrogen (C-H bonds), carbon atoms (C-C bonds), carbon and oxygen atoms (C-O bonds), and carbon and nitrogen (C-N bonds). Depending on the nature of the atoms, a covalent bond can be a single, double, or triple bond, i.e., a covalent bond can involve the sharing of one, two, or three electron pairs.

[0124] As used herein, the terms "ion," "ionic bond," and "ionic bonding" refer to a type of chemical bond involving Coulombic attraction between adjacent oppositely charged atoms (ie, ions).

[0125] Modified polysaccharide polymers

[0126] The polysaccharide polymers described herein are covalently modified with a covalent photoactive crosslinking agent. In one embodiment, the polysaccharide polymer can be a linear, branched or crosslinked polysaccharide polymer, or a polysaccharide polymer with a selected molecular weight range, degree of polymerization, viscosity or melt flow rate. Branched polysaccharide polymers can include one or more of the following types: star polymers, comb polymers, brush polymers, dendritic polymers, graft-copolymers, ladder polymers and dendritic polymers. In some embodiments, the branched polysaccharide polymer is a star polymer. In some embodiments, the branched polysaccharide polymer is a comb polymer. In some embodiments, the branched polysaccharide polymer is a brush polymer. In some embodiments, the branched polysaccharide polymer is a dendritic polymer. In some embodiments, the branched polysaccharide polymer is a graft-(co)polymer. In some embodiments, the branched polysaccharide polymer is a ladder polymer. In some embodiments, the branched polysaccharide polymer is a dendritic polymer. The polysaccharide polymer can be a thermosensitive polymer, such as a gel (e.g., a gel that becomes solid or liquid when exposed to heat or a certain temperature) or a photocrosslinkable polymer. In some embodiments, the polysaccharide polymer is a photocrosslinkable polymer. In some embodiments, the polysaccharide polymer can be biodegradable, for example, contain unstable bonds, or can be dissociated by enzymes (such as lyases). In some embodiments, the polysaccharide polymer is composed of a single type of repeating monomer units. In other embodiments, the polysaccharide polymer is composed of different types of repeating monomer units (for example, two types of repeating monomer units, three types of repeating monomer units, for example, polymer blends). In some embodiments, the polysaccharide polymer can be composed of mannuronic acid and guluronic acid monomers.

[0127] In some embodiments, the polysaccharide polymer is a naturally occurring or synthetic polymer. In some embodiments, the polysaccharide polymer is a naturally occurring polysaccharide or a synthetic polysaccharide. In one embodiment, the polysaccharide polymer is cellulose, such as carboxymethyl cellulose. In one embodiment, the polysaccharide polymer is polylactic acid, polyglycoside, or polycaprolactone. In one embodiment, the polysaccharide polymer is a hyaluronate, such as sodium hyaluronate. In one embodiment, the polysaccharide polymer is collagen, elastin, or gelatin. In one embodiment, the polysaccharide polymer is chitin.

[0128] In some embodiments, polysaccharide polymer is the polymer forming hydrogel.The polymer forming hydrogel comprises the hydrophilic structure that enables it to hold a large amount of water in three-dimensional network.The polymer forming hydrogel may include the polymer forming homopolymer hydrogel capsule, copolymer hydrogel capsule or polypolymer interpenetrating polymer hydrogel capsule, and can be amorphous, semi-crystalline or crystalline in nature, for example, as described in Ahmed (2015) J Adv Res 6:105-121.Exemplary polymer forming hydrogel includes protein (for example, collagen), gelatin, polysaccharide (for example, starch, alginate, hyaluronate, agarose) and synthetic polysaccharide.

[0129] Exemplary polysaccharide polymers include alginate, agar, agarose, carrageenan, hyaluronate, pullulan, glycogen, gelatin, cellulose, amylose, chitin, chitosan or its derivatives or variants, for example, as described in Laurienzo (2010), Mar Drugs 9:2435-65. Polysaccharide polymers can include heparin, chondroitin sulfate, dermatan, dextran or carboxymethyl cellulose. In some embodiments, the polysaccharide polymer is a cell surface polysaccharide.

[0130] In some embodiments, the polysaccharide polymer is alginate.Alginate is the polysaccharide being made up of β-D-mannuronic acid (M) and α-L-guluronic acid (G).In some embodiments, alginate is high guluronic acid (G) alginate, and comprises and is greater than about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or more guluronic acid (G).In some embodiments, alginate is high mannuronic acid (M) alginate, and comprises and is greater than about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or more mannuronic acid (M).In some embodiments, the ratio of M:G is about 1.In some embodiments, the ratio of M:G is less than 1.In some embodiments, the ratio of M:G is greater than 1.In some embodiments, alginate has the approximate molecular weight of <75 kDa and optionally the G:M ratio of ≥1.5. In some embodiments, the alginate has an approximate molecular weight of 75 kDa to 150 kDa and optionally a G:M ratio of > 1.5. In some embodiments, the alginate has an approximate molecular weight of 150 to 250 kDa and optionally a G:M ratio of > 1.5.

[0131] The polysaccharide polymer (for example, any polymer described herein, for example any alginate described herein) or its pharmaceutically acceptable salt comprising the sugar moiety with formula (I) structure can be modified on one or more monomeric units. In some embodiments, at least 0.5% of the sugar monomer in the polysaccharide polymer has the structure of formula (I) (for example, at least 1%, 2.5%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or more sugar monomer has the structure of formula (I)). In some embodiments, 0.5% to 50%, 10% to 90%, 10% to 50% or 25% to 75% of the sugar monomer of the polysaccharide polymer has the structure of formula (I). In some embodiments, 1% to 20% of the sugar monomer of the polysaccharide polymer has the structure of formula (I). In some embodiments, 1% to 10% of the sugar monomer of the polysaccharide polymer has the structure of formula (I). In some embodiments, 1% to 50% of the sugar monomers have the structure of formula (I).

[0132] In some embodiments, the polysaccharide polymer (when comprising sugar monomers having a structure of Formula I) comprises (compared to the unmodified polymer) an increase in N% of at least 0.1%, 0.2%, 0.5%, 1.0%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% N by weight, where N% is determined by elemental analysis and corresponds to the amount of compound of Formula I in the modified polymer.

[0133] In some embodiments, the polysaccharide polymer (when comprising sugar monomers having a structure of Formula I) comprises (compared to the unmodified polymer) an increase in N% of 0.1 wt% to 10 wt% N, where N% is determined by elemental analysis and corresponds to the amount of compound of Formula I in the modified polymer.

[0134] In some embodiments, the polysaccharide polymer (when comprising sugar monomers having a structure of Formula (I)) comprises an increase in N% (compared to the unmodified polymer) of 0.1 wt% to 2 wt% N, where N% is determined by elemental analysis and corresponds to the amount of compound of Formula I in the modified polymer.

[0135] In some embodiments, the polysaccharide polymer (when comprising sugar monomers having a structure of Formula (I)) comprises (compared to the unmodified polymer) an increase in N% of 2 to 4 wt% N, where N% is determined by elemental analysis and corresponds to the amount of compound of Formula I in the modified polymer.

[0136] In some embodiments, the polysaccharide polymer (when comprising sugar monomers having a structure of Formula (I)) comprises (compared to the unmodified polymer) an increase in N% of 4 to 8 wt% N, where N% is determined by elemental analysis and corresponds to the amount of compound of Formula I in the modified polymer.

[0137] In some embodiments, any polysaccharide polymer described herein (e.g., alginate) comprises a sugar monomer having one or more of formula (Ia), (Ib), (Ic), (Id), (Ie), (If), or a pharmaceutically acceptable salt thereof. In some embodiments, the polysaccharide polymer comprises a sugar monomer having a structure of formula (II-a). In some embodiments, the polymer is modified with a compound of formula (II-b). In some embodiments, the polysaccharide polymer comprises a sugar monomer having a structure of formula (II-c). In some embodiments, the polysaccharide polymer comprises a sugar monomer having a structure of formula (II-d). In some embodiments, the polysaccharide polymer comprises a sugar monomer having a structure of formula (II-e). In some embodiments, the polysaccharide polymer comprises a sugar monomer having a structure of formula (II-f).

[0138] In some embodiments, the polysaccharide polymer (e.g., alginate) is modified with a compound shown in Table 3. In some embodiments, the polymer (e.g., alginate) modified with a compound of formula (I) is not a modified polymer described in any of WO2012 / 112982, WO2012 / 167223, WO2014 / 153126, WO2016 / 187225, WO2016 / 019391, WO2017 / 075630, WO 2017 / 075631, WO 2018 / 067615, WO 2019 / 169333, and US 2016-0030359.

[0139] Defibrotizing compounds

[0140] In some embodiments, the polysaccharide polymers described herein further comprise at least one defibroting compound of formula (I):

[0141] (I),

[0142] or a pharmaceutically acceptable salt thereof, wherein:

[0143] A is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -O-, -C(O)O-, -C(O)-, -OC(O)-, -N(R C )–、–N(R C )C(O)–、–C(O)N(R C )–、-N(R C )C(O)(C1-C6-alkylene)–、-N(R C )C(O)(C1-C6-alkenylene)–, –N(R C )N(R D )–、–NCN–、–C(=N(R C )(R D ))O–, –S–, –S(O) x –,–OS(O) x –、–N(R C )S(O) x –, –S(O) x N(R C )–、–P(R F ) y –、–Si(OR A )2–、–Si(R G )(OR A )–、–B(OR A )— or a metal, each of which is optionally linked to an attachment group (such as an attachment group described herein) and optionally surrounded by one or more R 1 Replace; L 1 and L 3 Each of the R 2 Replace; L 2 is a bond; M is absent, alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally replaced by one or more R 3 substituted; P is absent, cycloalkyl, heterocyclyl or heteroaryl, each of which is optionally substituted by one or more R 4 Substituted; Z is hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, –OR A ,–C(O)R A 、–C(O)OR A 、–C(O)N(R C )(R D ),–N(R C )C(O)R A , cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally replaced by one or more R5 Replace; each R A 、R B 、R C 、R D 、R E 、R F and R G are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, azido, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally replaced by one or more R 6 Substitute; or R C and R D Together with the nitrogen atom to which it is attached, it forms a 6 substituted ring (eg, 5- to 7-membered ring); each R 1 、R 2 、R 3 、R 4 、R 5 and R 6 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 、S(O) x R E1 、–OS(O) x R E1 ,–N(R C1 )S(O) x R E1 ,–S(O) x N(R C1 )(R D1 ),–P(R F1 ) y , cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally replaced by one or more R 7 Replace; each R A1 、R B1 、R C1 、R D1 、R E1 and R F1are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally replaced by one or more R 7 Replace; each R 7 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, oxo, hydroxy, cycloalkyl, or heterocyclyl; x is 1 or 2; and y is 2, 3, or 4.

[0144] In some embodiments, the compound of formula (I) is a compound of formula (Ia): (Ia),

[0145] or a pharmaceutically acceptable salt thereof, wherein: A is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -O-, -C(O)O-, -C(O)-, -OC(O)-, -N(R C )–、–N(R C )C(O)–、–C(O)N(R C )–、–N(R C )N(R D )–、N(R C )C(O)(C1-C6-alkylene)–、-N(R C )C(O)(C1-C6-alkenylene)–, –NCN–, –C(=N(R C )(R D ))O–, –S–, –S(O) x –,–OS(O) x –、–N(R C )S(O) x –, –S(O) x N(R C )–、–P(R F ) y –、–Si(OR A )2–、–Si(R G )(OR A )–、–B(OR A )— or a metal, each of which is optionally linked to an attachment group (such as an attachment group described herein) and optionally surrounded by one or more R 1 Replace; L 1 and L 3 Each of the R 2 Replace; L 2is a bond; M is absent, alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally replaced by one or more R 3 substituted; P is optionally substituted with one or more R 4 substituted heteroaryl; Z is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally substituted by one or more R 5 Replace; each R A 、R B 、R C 、R D 、R E 、R F and R G are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, azido, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally replaced by one or more R 6 Substitute; or R C and R D Together with the nitrogen atom to which it is attached, it forms a 6 substituted ring (eg, 5- to 7-membered ring); each R 1 、R 2 、R 3 、R 4 、R 5 and R 6 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 、S(O) x R E1 、–OS(O) x R E1 ,–N(R C1 )S(O) x R E1 ,–S(O) x N(R C1 )(R D1 ),–P(R F1 ) y, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally replaced by one or more R 7 Replace; each R A1 、R B1 、R C1 、R D1 、R E1 and R F1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally replaced by one or more R 7 Replace; each R 7 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, oxo, hydroxy, cycloalkyl, or heterocyclyl; x is 1 or 2; and y is 2, 3, or 4.

[0146] In some embodiments, for Formula (I) and (Ia), A is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -O-, -C(O)O-, -C(O)-, -OC(O)-, -N(R C )C(O)-、-N(R C )C(O)(C1-C6-alkylene)-, -N(R C )C(O)(C1-C6-alkenylene)-or-N(R C )-. In some embodiments, A is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -O-, -C(O)O-, -C(O)-, -OC(O)-, or -N(R C )-. In some embodiments, A is alkyl, alkenyl, alkynyl, heteroalkyl, -O-, -C(O)O-, -C(O)-, -OC(O)-, or -N(R C )-. In some embodiments, A is alkyl, -O-, -C(O)O-, -C(O)-, -OC(O), or -N(R C )-. In some embodiments, A is -N(R C )C(O)-、-N(R C )C(O)(C1-C6-alkylene)-or-N(R C )C(O)(C1-C6-alkenylene)-. In some embodiments, A is -N(R C )-. In some embodiments, A is -N(R C )-, and R C and R D In some embodiments, A is -NH-. In some embodiments, A is -N(RC )C(O)(C1-C6-alkylene)-, wherein the alkylene is replaced by R 1 In some embodiments, A is -N(R C )C(O)(C1-C6-alkylene)-, and R 1 is alkyl (e.g., methyl). In some embodiments, A is -NHC(O)C(CH3)2-. In some embodiments, A is -N(R C )C(O)(methylene)-, and R 1 is alkyl (e.g., methyl). In some embodiments, A is -NHC(O)CH(CH3)-. In some embodiments, A is -NHC(O)C(CH3)-.

[0147] In some embodiments, for Formula (I) and (Ia), L 1 is a bond, alkyl, or heteroalkyl. In some embodiments, L 1 is a bond or an alkyl group. In some embodiments, L 1 In some embodiments, L 1 In some embodiments, L 1 is C1-C6 alkyl. In some embodiments, L 1 Is -CH2-, -CH(CH3)-, -CH2CH2CH2 or -CH2CH2-. In some embodiments, L 1 It is -CH2- or -CH2CH2-.

[0148] In some embodiments, for Formula (I) and (Ia), L 3 is a bond, alkyl, or heteroalkyl. In some embodiments, L 3 In some embodiments, L 3 In some embodiments, L 3 It is C1-C 12 In some embodiments, L 3 is C1-C6 alkyl. In some embodiments, L 3 In some embodiments, L 3 In some embodiments, L 3 is optionally replaced by one or more R 2 (e.g., oxo) substituted C1-C 12 In some embodiments, L 3 is optionally replaced by one or more R 2 In some embodiments, L 3is -C(O)OCH2-, -CH2(OCH2CH2)2-, -CH2(OCH2CH2)3-, CH2CH2O-, or -CH2O-. In some embodiments, L 3 It is -CH2O-.

[0149] In some embodiments, for Formula (I) and Formula (Ia), M is absent, alkyl, heteroalkyl, aryl, or heteroaryl. In some embodiments, M is heteroalkyl, aryl, or heteroaryl. In some embodiments, M is absent. In some embodiments, M is alkyl (e.g., C1-C6 alkyl). In some embodiments, M is -CH2-. In some embodiments, M is heteroalkyl (e.g., C1-C6 heteroalkyl). In some embodiments, M is (-OCH2CH2-)z, wherein z is an integer selected from 1 to 10. In some embodiments, z is an integer selected from 1 to 5. In some embodiments, M is -OCH2CH2-, (-OCH2CH2-)2, (-OCH2CH2-)3, (-OCH2CH2-)4, or (-OCH2CH2-)5. In some embodiments, M is -OCH2CH2-, (-OCH2CH2-)2, (-OCH2CH2-)3, or (-OCH2CH2-)4. In some embodiments, M is (-OCH2CH2-)3. In some embodiments, for Formula (I) and Formula (Ia), M is absent, alkyl, heteroalkyl, aryl, or heteroaryl. In some embodiments, M is heteroalkyl, aryl, or heteroaryl. In some embodiments, M is absent. In some embodiments, M is alkyl (e.g., C1-C6 alkyl). In some embodiments, M is -CH2-. In some embodiments, M is heteroalkyl (e.g., C1-C6 heteroalkyl). In some embodiments, M is (-OCH2CH2-)z, wherein z is an integer selected from 1 to 10. In some embodiments, z is an integer selected from 1 to 5. In some embodiments, M is -OCH2CH2-, (-OCH2CH2-)2, (-OCH2CH2-)3, (-OCH2CH2-)4, or (-OCH2CH2-)5. In some embodiments, M is -OCH2CH2-, (-OCH2CH2-)2, (-OCH2CH2-)3, or (-OCH2CH2-)4. In some embodiments, M is (-OCH2CH2-)3. In some embodiments, M is aryl. In some embodiments, M is phenyl. In some embodiments, M is unsubstituted phenyl. In some embodiments, M is In some embodiments, M is replaced by R 7 (For example, 1 R 7 In some embodiments, M is In some embodiments, R 7 It's CF3.

[0150] In some embodiments, for Formula (I) and Formula (Ia), P is absent, heterocyclic, or heteroaryl. In some embodiments, P is absent. In some embodiments, for Formula (I) and (Ia), P is a tricyclic, bicyclic, or monocyclic heteroaryl. In some embodiments, P is a monocyclic heteroaryl. In some embodiments, P is a nitrogen-containing heteroaryl. In some embodiments, P is a monocyclic nitrogen-containing heteroaryl. In some embodiments, P is a 5-membered heteroaryl. In some embodiments, P is a 5-membered nitrogen-containing heteroaryl. In some embodiments, P is tetrazolyl, imidazolyl, pyrazolyl, or triazolyl, pyrrolyl, oxazolyl, or thiazolyl. In some embodiments, P is tetrazolyl, imidazolyl, pyrazolyl, or triazolyl or pyrrolyl. In some embodiments, P is imidazolyl. In some embodiments, P is In some embodiments, P is triazolyl. In some embodiments, P is 1,2,3-triazolyl. In some embodiments, P is .

[0151] In some embodiments, P is a heterocyclyl. In some embodiments, P is a 5-membered heterocyclyl or a 6-membered heterocyclyl. In some embodiments, P is an imidazolidinone. In some embodiments, P is In some embodiments, P is thiomorpholinyl-1,1-dioxide.

[0152] In some embodiments, P is .

[0153] In some embodiments, for Formula (I) and Formula (Ia), Z is alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl. In some embodiments, Z is heterocyclyl. In some embodiments, Z is a monocyclic or bicyclic heterocyclyl. In some embodiments, Z is an oxygen-containing heterocyclyl. In some embodiments, Z is a 4-membered heterocyclyl, a 5-membered heterocyclyl, or a 6-membered heterocyclyl. In some embodiments, Z is a 6-membered heterocyclyl. In some embodiments, Z is a 6-membered oxygen-containing heterocyclyl. In some embodiments, Z is tetrahydropyranyl. In some embodiments, Z is , or In some embodiments, Z is a 4-membered oxygen-containing heterocyclic group. In some embodiments, Z is .

[0154] In some embodiments, Z is a bicyclic oxygen-containing heterocyclic group. In some embodiments, Z is a phthalic anhydride group. In some embodiments, Z is a sulfur-containing heterocyclic group. In some embodiments, Z is a 6-membered sulfur-containing heterocyclic group. In some embodiments, Z is a 6-membered heterocyclic group containing a nitrogen atom and a sulfur atom. In some embodiments, Z is a thiomorpholinyl-1,1-dioxyl group. In some embodiments, Z is In some embodiments, Z is a nitrogen-containing heterocyclic group. In some embodiments, Z is a 6-membered nitrogen-containing heterocyclic group. In some embodiments, Z is .

[0155] In some embodiments, Z is a bicyclic heterocyclyl. In some embodiments, Z is optionally substituted with one or more R 5 In some embodiments, Z is 2-oxa-7-azaspiro[3.5]nonyl. In some embodiments, Z is In some embodiments, Z is 1-oxa-3,8-diazaspiro[4.5]decan-2-one. In some embodiments, Z is .

[0156] In some embodiments, for Formula (I) and (Ia), Z is aryl. In some embodiments, Z is monocyclic aryl. In some embodiments, Z is phenyl. In some embodiments, Z is (e.g., surrounded by 1 R 5 In some embodiments, Z is a monosubstituted phenyl group, wherein one R 5 In some embodiments, Z is a monosubstituted phenyl group, wherein one R 5 In some embodiments, Z is a monosubstituted phenyl group, wherein one R 5 In some embodiments, Z is a monosubstituted phenyl group, wherein one R 5 In some embodiments, Z is a monosubstituted phenyl group, wherein one R 5 In some embodiments, Z is a monosubstituted phenyl group, wherein one R 5 In some embodiments, Z is a monosubstituted phenyl group, wherein one R 5 In some embodiments, Z is a monosubstituted phenyl group, wherein one R 5 Located in the opposite position.

[0157] In some embodiments, for Formula (I) and (Ia), Z is alkyl. In some embodiments, Z is C1-C 12 In some embodiments, Z is C1-C 10In some embodiments, Z is C1-C8 alkyl. In some embodiments, Z is surrounded by 1-5 R 5 In some embodiments, Z is replaced by 1 R 5 In some embodiments, Z is replaced by 1 R 5 Substituted C1-C8 alkyl, wherein R 5 is alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 、-C(O)R B1 、-OC(O)R B1 or -N(R C1 )(R D1 In some embodiments, Z is replaced by 1 R 5 Substituted C1-C8 alkyl, wherein R 5 Yes-OR A1 or-C(O)OR A1 In some embodiments, Z is replaced by 1 R 5 Substituted C1-C8 alkyl, wherein R 5 Yes-OR A1 Or -C(O)OH. In some embodiments, Z is -CH3.

[0158] In some embodiments, for Formula (I) and (Ia), Z is heteroalkyl. In some embodiments, Z is C1-C 12 In some embodiments, Z is C1-C 10 In some embodiments, Z is C1-C8 heteroalkyl. In some embodiments, Z is C1-C6 heteroalkyl. In some embodiments, Z is optionally substituted by one or more R 5 In some embodiments, Z is substituted with 1-5 R 5 Substituted nitrogen and sulfur containing heteroalkyl. In some embodiments, Z is N-methyl-2-(methylsulfonyl)eth-1-amino (aminyl).

[0159] In some embodiments, Z is -OR A or-C(O)OR A In some embodiments, Z is -OR A (e.g., -OH or -OCH3). In some embodiments, Z is -OCH3. In some embodiments, Z is -C(O)OR A (e.g., -C(O)OH).

[0160] In some embodiments, Z is hydrogen.

[0161] In some embodiments, L 2 is a key and P and L 3 In some embodiments, L 2 is a bond, P is a heteroaryl, L 3 is a bond, and Z is hydrogen. In some embodiments, P is heteroaryl, L 3 is heteroalkyl, and Z is alkyl.

[0162] In some embodiments, the compound of formula (I) is a compound of formula (Ib):

[0163] (Ib),

[0164] or a pharmaceutically acceptable salt thereof, wherein ring M 1 is cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally substituted by 1-5 R 3 Substitution; Ring Z 1 is optionally replaced by 1-5 R 5 Substituted cycloalkyl, heterocyclyl, aryl or heteroaryl; R 2a 、R 2b 、R 2c and R 2d Each of is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, nitro, amino, cycloalkyl, heterocyclyl, aryl or heteroaryl, or R 2a and R 2b or R 2c and R 2d Each of the groups together form an oxo group; X is absent, N(R 10 )(R 11 ), O or S; R C is hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted by 1-6 R 6 Replace; each R 3 、R 5 and R 6 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、-C(O)OR A1 、-C(O)R B1 、-OC(O)R B1 、-N(R C1 )(R D1 )、-N(R C1 )C(O)R B1 、-C(O)N(R C1 ), SRE1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; R 10 and R 11 Each of which is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, -C(O)OR A1 、-C(O)R B1 、-OC(O)R B1 、-C(O)N(R C1 ), cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 Replace; each R 7 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxy, cycloalkyl or heterocyclyl; each m and n is independently 1, 2, 3, 4, 5 or 6; and “ " refers to a connection to an attachment group or polymer described herein. In some embodiments, for each R 3 and R 5 , each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally and independently substituted with halo, oxo, cyano, cycloalkyl or heterocyclyl.

[0165] In some embodiments, the compound of formula (Ib) is a compound of formula (Ibi):

[0166] (Ibi),

[0167] or a pharmaceutically acceptable salt thereof, wherein ring M 2 is optionally replaced by one or more R 3 Substituted aryl or heteroaryl; Ring Z 2 is a cycloalkyl, heterocyclyl, aryl or heteroaryl group; R 2a 、R 2b 、R 2c and R 2d Each of R is independently hydrogen, alkyl or heteroalkyl, or R 2a and R 2b or R 2c and R 2d Each of together form an oxo group; X is absent, O or S; each R 3 and R 5 are independently alkyl, heteroalkyl, halogen, oxo, -ORA1 、-C(O)OR A1 or -C(O)R B1 , wherein each alkyl and heteroalkyl is optionally substituted with halogen; or two R 5 Together with ring Z 2 fused 5-6 membered ring; each R A1 and R B1 is independently hydrogen, alkyl or heteroalkyl; m and n are each independently 1, 2, 3, 4, 5 or 6; p is 0, 1, 2, 3, 4, 5 or 6; and “ ” refers to a connection to an attachment group or polymer described herein.

[0168] In some embodiments, the compound of formula (Ibi) is a compound of formula (Ib-ii):

[0169] (Ib-ii),

[0170] or a pharmaceutically acceptable salt thereof, wherein ring Z 2 is a cycloalkyl, heterocyclyl, aryl or heteroaryl group; R 2c and R 2d Each of R is independently hydrogen, alkyl or heteroalkyl, or R 2c and R 2d together to form an oxo group; each R 3 and R 5 are independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 or -C(O)R B1 , wherein each alkyl and heteroalkyl is optionally substituted with halogen; each R A1 and R B1 is independently hydrogen, alkyl or heteroalkyl; each of p and q is independently 0, 1, 2, 3, 4, 5 or 6; and “ ” refers to a connection to an attachment group or polymer described herein.

[0171] In some embodiments, the compound of formula (I) is a compound of formula (Ic):

[0172] (Ic),

[0173] or a pharmaceutically acceptable salt thereof, wherein ring Z 2 is a cycloalkyl, heterocyclyl, aryl or heteroaryl group; R 2c and R 2d Each of R is independently hydrogen, alkyl or heteroalkyl, or R 2c and R 2d together to form an oxo group; each R3 and R 5 are independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 or -C(O)R B1 , wherein each alkyl and heteroalkyl is optionally substituted with halogen; each R A1 and R B1 is independently hydrogen, alkyl or heteroalkyl; m is 1, 2, 3, 4, 5 or 6; each of p and q is independently 0, 1, 2, 3, 4, 5 or 6; and “ ” refers to a connection to an attachment group or polymer described herein.

[0174] In some embodiments, the compound of formula (I) is a compound of formula (Id):

[0175] (Id),

[0176] or a pharmaceutically acceptable salt thereof, wherein ring Z 2 is cycloalkyl, heterocyclyl, aryl or heteroaryl; X is absent, O or S; R 2a 、R 2b 、R 2c and R 2d Each of R is independently hydrogen, alkyl or heteroalkyl, or R 2a and R 2b or R 2c and R 2d Each of the R 5 are independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 or -C(O)R B1 , wherein each alkyl and heteroalkyl is optionally substituted with halogen; each R A1 and R B1 is independently hydrogen, alkyl, or heteroalkyl; each of m and n is independently 1, 2, 3, 4, 5, or 6; p is 0, 1, 2, 3, 4, 5, or 6; and “ ” refers to a connection to an attachment group or polymer described herein.

[0177] In some embodiments, the compound of formula (I) is a compound of formula (Ie):

[0178] (Ie),

[0179] or a pharmaceutically acceptable salt thereof, wherein ring Z 2 is cycloalkyl, heterocyclyl, aryl or heteroaryl; X is absent, O or S; R 2a 、R2b 、R 2c and R 2d Each of R is independently hydrogen, alkyl or heteroalkyl, or R 2a and R 2b or R 2c and R 2d Each of the R 5 are independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 or -C(O)R B1 ; Each R A1 and R B1 is independently hydrogen, alkyl, or heteroalkyl; each of m and n is independently 1, 2, 3, 4, 5, or 6; p is 0, 1, 2, 3, 4, 5, or 6; and “ ” refers to a connection to an attachment group or polymer described herein.

[0180] In some embodiments, the compound of formula (I) is a compound of formula (If):

[0181] (If),

[0182] or a pharmaceutically acceptable salt thereof, wherein M is optionally replaced by one or more R 3 substituted alkyl; Ring P is optionally substituted with one or more R 4 Substituted heteroaryl; L 3 is optionally replaced by one or more R 2 substituted alkyl or heteroalkyl; Z is alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally replaced by one or more R 5 Replacement; R 2a and R 2b Each of R is independently hydrogen, alkyl or heteroalkyl, or R 2a and R 2b together to form an oxo group; each R 2 、R 3 、R 4 and R 5 are independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 or -C(O)R B1 ; Each R A1 and R B1 is independently hydrogen, alkyl or heteroalkyl; n is independently 1, 2, 3, 4, 5 or 6; and “ ” refers to a connection to an attachment group or polymer described herein.

[0183] In some embodiments, the compound of formula (I) is a compound of formula (II):

[0184] (II),

[0185] or a pharmaceutically acceptable salt thereof, wherein M is a bond, an alkyl group, or an aryl group, wherein the alkyl group and the aryl group are optionally replaced by one or more R 3 Replace; L 3 is optionally replaced by one or more R 2 substituted alkyl or heteroalkyl; Z is hydrogen, alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl or -OR A , wherein alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally replaced by one or more R 5 Replacement; R A is hydrogen; R 2a and R 2b Each of R is independently hydrogen, alkyl or heteroalkyl, or R 2a and R 2b together to form an oxo group; each R 2 、R 3 and R 5 are independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 or -C(O)R B1 ; Each R A1 and R B1 is independently hydrogen, alkyl or heteroalkyl; n is independently 1, 2, 3, 4, 5 or 6; and “ ” refers to a connection to an attachment group or polymer described herein.

[0186] In some embodiments, the compound of formula (II) is a compound of formula (II-a):

[0187] (II-a),

[0188] or a pharmaceutically acceptable salt thereof, wherein L 3 is alkyl or heteroalkyl, each of which is optionally replaced by one or more R 2 Substituted; Z is hydrogen, alkyl, heteroalkyl or -OR A , wherein alkyl and heteroalkyl are optionally replaced by one or more R 5 Replacement; R 2a and R 2b Each of R is independently hydrogen, alkyl or heteroalkyl, or R 2a and R 2b together to form an oxo group; each R 2 、R 3 and R 5are independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 or -C(O)R B1 ; R A is hydrogen; each R A1 and R B1 is independently hydrogen, alkyl or heteroalkyl; n is independently 1, 2, 3, 4, 5 or 6; and “ ” refers to a connection to an attachment group or polymer described herein.

[0189] In some embodiments, the compound of formula (I) is a compound of formula (III):

[0190] (III),

[0191] or a pharmaceutically acceptable salt thereof, wherein Z 1 is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally substituted by 1-5 R 5 Replacement; R 2a 、R 2b 、R 2c and R 2d Each of is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, nitro, amino, cycloalkyl, heterocyclyl, aryl or heteroaryl; or R 2a and R 2b or R 2c and R 2d Together they form an oxo group; R C is hydrogen, alkyl, alkenyl, alkynyl or heteroalkyl, wherein each of the alkyl, alkenyl, alkynyl or heteroalkyl is optionally substituted with 1-6 R 6 Replacement; R 3 、R 5 and R 6 Each of which is independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 or -C(O)R B1 ; Each R A1 and R B1 is independently hydrogen, alkyl or heteroalkyl; m and n are each independently 1, 2, 3, 4, 5 or 6; q is an integer from 0 to 25; and “ ” refers to a connection to an attachment group or polymer described herein.

[0192] In some embodiments, the compound of formula (III) is a compound of formula (III-a):

[0193] (III-a),

[0194] or a pharmaceutically acceptable salt thereof, wherein ring Z 2 is cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally substituted by 1-5 R 5 Replacement; R 2a 、R 2b 、R 2c and R 2d Each of R is independently hydrogen, alkyl, heteroalkyl, halo; or 2a and R 2b or R 2c and R 2d Together they form an oxo group; R 3 and R 5 Each of which is independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 or -C(O)R B1 ; Each R A1 and R B1 is independently hydrogen, alkyl or heteroalkyl; m and n are each independently 1, 2, 3, 4, 5 or 6; o and p are each independently 0, 1, 2, 3, 4 or 5; q is an integer from 0 to 25; and “ ” refers to a connection to an attachment group or polymer described herein.

[0195] In some embodiments, the compound of formula (III-a) is a compound of formula (III-b):

[0196] (III-b),

[0197] or a pharmaceutically acceptable salt thereof, wherein ring Z 2 is cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally substituted by 1-5 R 5 Replacement; R 2a 、R 2b 、R 2c and R 2d Each of R is independently hydrogen, alkyl, heteroalkyl, halo; or 2a and R 2b or R 2c and R 2d Together they form an oxo group; R 3 and R 5 Each of which is independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 or -C(O)R B1 ; Each R A1 and R B1is independently hydrogen, alkyl or heteroalkyl; m and n are each independently 1, 2, 3, 4, 5 or 6; o and p are each independently 0, 1, 2, 3, 4 or 5; q is an integer from 0 to 25; and “ ” refers to a connection to an attachment group or polymer described herein.

[0198] In some embodiments, the compound of formula (III-a) is a compound of formula (III-c):

[0199] (III-c),

[0200] or a pharmaceutically acceptable salt thereof, wherein X is C(R')(R"), N(R') or S(O) x ; R' and R" are each independently hydrogen, alkyl, halogen or cycloalkyl; R 2a 、R 2b 、R 2c and R 2d Each of R is independently hydrogen, alkyl, heteroalkyl or halo; or 2a and R 2b or R 2c and R 2d Together they form an oxo group; R 3 and R 5 Each of which is independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 or -C(O)R B1 ; Each R A1 and R B1 is independently hydrogen, alkyl or heteroalkyl; m and n are each independently 1, 2, 3, 4, 5 or 6; p is 0, 1, 2, 3, 4 or 5; q is an integer from 0 to 25; x is 0, 1 or 2; and “ ” refers to a connection to an attachment group or polymer described herein.

[0201] In some embodiments, the compound of formula (III-c) is a compound of formula (III-d):

[0202] (III-d),

[0203] or a pharmaceutically acceptable salt thereof, wherein X is C(R')(R"), N(R') or S(O) x ; R' and R" are each independently hydrogen, alkyl, halogen or cycloalkyl; R 2a 、R 2b 、R 2c and R 2d Each of R is independently hydrogen, alkyl, heteroalkyl or halo; or2a and R 2b or R 2c and R 2d Together they form an oxo group; R 3 and R 5 Each of which is independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 or -C(O)R B1 ; Each R A1 and R B1 is independently hydrogen, alkyl or heteroalkyl; m and n are each independently 1, 2, 3, 4, 5 or 6; p is 0, 1, 2, 3, 4 or 5; q is an integer from 0 to 25; x is 0, 1 or 2; and “ ” refers to a connection to an attachment group or polymer described herein.

[0204] In some embodiments, the compound of formula (I) is a compound of formula (III-e):

[0205] (III-e),

[0206] or a pharmaceutically acceptable salt thereof, wherein Z 1 is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally substituted by 1-5 R 5 Replacement; R 2a 、R 2b 、R 2c and R 2d Each of is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, nitro, amino, cycloalkyl, heterocyclyl, aryl or heteroaryl; or R 2a and R 2b or R 2c and R 2d Each of the R C is hydrogen, alkyl, alkenyl, alkynyl or heteroalkyl, wherein each of the alkyl, alkenyl, alkynyl or heteroalkyl is optionally substituted with 1-6 R 6 Replacement; R 3 、R 5 and R 6 Each of which is independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 or -C(O)R B1 ; Each R 12 are independently deuterium, alkyl, heteroalkyl, haloalkyl, halo, cyano, nitro, or amino; each R A1 and R B1is independently hydrogen, alkyl or heteroalkyl; m and n are each independently 1, 2, 3, 4, 5 or 6; q is an integer from 0 to 25; w is 0 or 1; and " ” refers to a connection to an attachment group or polymer described herein.

[0207] In some embodiments, the compound of formula (I) is a compound of formula (III-f):

[0208] (III-f),

[0209] or a pharmaceutically acceptable salt thereof, wherein ring Z 1 is cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally substituted by 1-5 R 5 Replacement; R 2a 、R 2b 、R 2c and R 2d Each of R is independently hydrogen, alkyl, heteroalkyl, halo; or 2a and R 2b or R 2c and R 2d Together they form an oxo group; R C is hydrogen, alkyl, alkenyl, alkynyl or heteroalkyl, wherein each of the alkyl, alkenyl, alkynyl or heteroalkyl is optionally substituted with 1-6 R 6 Replacement; R 3 、R 5 and R 6 Each of which is independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 or -C(O)R B1 ; Each R 12 are independently deuterium, alkyl, heteroalkyl, haloalkyl, halogen, cyano, nitro or amino; each R A1 and R B1 is independently hydrogen, alkyl or heteroalkyl; m and n are each independently 1, 2, 3, 4, 5 or 6; o and p are each independently 0, 1, 2, 3, 4 or 5; q is an integer from 0 to 25; w is 0 or 1; and “ ” refers to a connection to an attachment group or polymer described herein.

[0210] In some embodiments, the compound of formula (I) is a compound of formula (III-g):

[0211] (III-g),

[0212] or a pharmaceutically acceptable salt thereof, wherein ring Z 1is cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally substituted by 1-5 R 5 Replacement; R C is hydrogen, alkyl, –N(R C )C(O)R B ,–N(R C )C(O)(C1-C6-alkyl) or –N(R C )C(O)(C1-C6-alkenyl), wherein each of the alkyl and alkenyl groups is optionally substituted with 1-6 R 6 Replacement; R 2a 、R 2b 、R 2c and R 2d Each of R is independently hydrogen or alkyl; or 2a and R 2b or R 2c and R 2d Together they form an oxo group; R 3 、R 5 and R 6 Each of is independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、–C(O)OR A1 or –C(O)R B1 ; R 12 is hydrogen, deuterium, alkyl, heteroalkyl, haloalkyl, halogen, cyano, nitro, or amino; each R A1 and R B1 is independently hydrogen, alkyl or heteroalkyl; m and n are each independently 1, 2, 3, 4, 5 or 6; q is an integer from 0 to 25; x is 0, 1 or 2; and “ ” refers to a connection to an attachment group or polymer described herein.

[0213] In some embodiments, the compound of formula (I) is a compound of formula (III-h):

[0214] (III-h),

[0215] or a pharmaceutically acceptable salt thereof, wherein R C is hydrogen, alkyl, -N(R C )C(O)R B 、-N(R C )C(O)(C1-C6-alkyl) or -N(R C )C(O)(C1-C6-alkenyl), wherein each of the alkyl and alkenyl groups is optionally substituted with 1-6 R 6 Replacement; R 2a 、R 2b 、R 2c and R 2dEach of R is independently hydrogen or alkyl; or 2a and R 2b or R 2c and R 2d Together they form an oxo group; R 3 、R 5 and R 6 Each of which is independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 or -C(O)R B1 ; R 12 is hydrogen, deuterium, alkyl, heteroalkyl, haloalkyl, halo, cyano, nitro, or amino; each R A1 and R B1 is independently hydrogen, alkyl or heteroalkyl; m and n are each independently 1, 2, 3, 4, 5 or 6; q is an integer from 0 to 25; x is 0, 1 or 2; z is 0, 1, 2, 3, 4, 5 or 6, and “ ” refers to a connection to an attachment group or polymer described herein.

[0216] In some embodiments, the compound of formula (I) is a compound of formula (III-i):

[0217] (III-i),

[0218] or a pharmaceutically acceptable salt thereof, wherein X is C(R')(R"), N(R') or S(O) x ; R' and R" are each independently hydrogen, alkyl or halogen; R C is hydrogen, alkyl, -N(R C )C(O)R B 、-N(R C )C(O)(C1-C6-alkyl) or -N(R C )C(O)(C1-C6-alkenyl), wherein each of the alkyl and alkenyl groups is optionally substituted with 1-6 R 6 Replacement; R 2a 、R 2b 、R 2c and R 2d Each of R is independently hydrogen or alkyl; or 2a and R 2b or R 2c and R 2d Together they form an oxo group; R 3 、R 5 and R 6 Each of which is independently alkyl, heteroalkyl, halogen, oxo, -OR A1 、-C(O)OR A1 or -C(O)RB1 ; R 12 is hydrogen, deuterium, alkyl, heteroalkyl, haloalkyl, halo, cyano, nitro, or amino; each R A1 and R B1 is independently hydrogen, alkyl or heteroalkyl; m and n are each independently 1, 2, 3, 4, 5 or 6; q is an integer from 0 to 25; x is 0, 1 or 2; z is 0, 1, 2, 3, 4, 5 or 6, and “ ” refers to a connection to an attachment group or polymer described herein.

[0219] In some embodiments, the compound is a compound of formula (I). In some embodiments, L 2 is a key and P and L 3 Does not exist independently.

[0220] In some embodiments, the compound is a compound of formula (Ia). In some embodiments of formula (II-a), L 2 is a bond, P is a heteroaryl, L 3 is a bond, and Z is hydrogen. In some embodiments, P is heteroaryl, L 3 is heteroalkyl, and Z is alkyl. In some embodiments, L 2 is a key and P and L 3 In some embodiments, L 2 is a bond, P is a heteroaryl, L 3 is a bond, and Z is hydrogen. In some embodiments, P is heteroaryl, L 3 is heteroalkyl, and Z is alkyl.

[0221] In some embodiments, the compound is a compound of formula (Ib). In some embodiments, P is absent, L 1 Yes -NHCH2, L 2 is a bond, M is an aryl group (e.g., phenyl), L 3 is -CH2O, and Z is a heterocyclic group (eg, a nitrogen-containing heterocyclic group such as thiomorpholinyl-1,1-dioxide).

[0222] In some embodiments of Formula (Ib), P is absent, L 1 Formula -NHCH2, L 2 is a key, M does not exist, L 3 is a bond, and Z is a heterocyclic group (eg, an oxygen-containing heterocyclic group such as tetrahydropyranyl, tetrahydrofuranyl, oxetanyl, or oxiranyl).

[0223] In some embodiments, the compound is of formula (Ibi). In some embodiments of formula (Ibi), R 2a and R2b Each of R is independently hydrogen or CH3, 2c and R 2d Each of the following is independently hydrogen, m is 1 or 2, n is 1, X is O, p is 0, M 2 is optionally replaced by one or more R 3 Substituted phenyl, R 3 is -CF3, and Z 2 is a heterocyclic group (for example, an oxygen-containing heterocyclic group such as tetrahydropyranyl, tetrahydrofuranyl, oxetanyl or oxiranyl).

[0224] In some embodiments, the compound is a compound of Formula (Ib-ii). In some embodiments of Formula (Ib-ii), R 2a 、R 2b 、R 2c and R 2d Each of is independently hydrogen, q is 0, p is 0, m is 1, and Z 2 is a heterocyclic group (eg, an oxygen-containing heterocyclic group such as tetrahydropyranyl).

[0225] In some embodiments, the compound is a compound of Formula (Ic). In some embodiments of Formula (Ic), R 2c and R 2d Each of the R is independently hydrogen, m is 1, p is 1, q is 0, R 5 is -CH3, and Z is a heterocyclic group (eg, a nitrogen-containing heterocyclic group such as piperazinyl).

[0226] In some embodiments, the compound is a compound of Formula (Id). In some embodiments of Formula (Id), R 2a 、R 2b 、R 2c and R 2d Each of wherein is independently hydrogen, m is 1, n is 3, X is O, p is 0, and Z is a heterocyclyl (eg, an oxygen-containing heterocyclyl such as tetrahydropyranyl, tetrahydrofuranyl, oxetanyl, or oxiranyl).

[0227] In some embodiments, the compound is of formula (If). In some embodiments of formula (If), R 2a and R 2b Each of the above is independently hydrogen, n is 1, M is -CH2-, P is a nitrogen-containing heteroaryl group (eg, imidazolyl), L 3 is -C(O)OCH2-, and Z is CH3.

[0228] In some embodiments, the compound is a compound of formula (II-a). In some embodiments of formula (II-a), R 2aand R 2b Each of the above is independently hydrogen, n is 1, q is 0, L 3 is -CH2(OCH2CH2)2, and Z is -OCH3.

[0229] In some embodiments of Formula (II-a), R 2a and R 2b Each of the L is independently hydrogen, n is 1, L 3 is a bond or –CH 2， and Z is hydrogen or –OH 。

[0230] In some embodiments, the compound is a compound of formula (III). In some embodiments of formula (III), R 2a 、R 2b 、R 2c and R 2d Each of the R is independently hydrogen, m is 1, n is 2, q is 3, p is 0, R C is hydrogen, and Z 1 is optionally replaced by R 5 Substituted heteroalkyl (e.g., -N(CH3)(CH2CH2)S(O)2CH3).

[0231] In some embodiments, the compound is a compound of formula (III-b). In some embodiments of formula (III-b), R 2a 、R 2b 、R 2c and R 2d Each of is independently hydrogen, m is 0, n is 2, q is 3, p is 0, and Z 2 Is 1 R 5 (e.g., -NH2) substituted aryl (e.g., phenyl).

[0232] In some embodiments, the compound is a compound of formula (III-b). In some embodiments of formula (III-b), R 2a 、R 2b 、R 2c and R 2d Each of the R is independently hydrogen, m is 1, n is 2, q is 3, p is 0, R C is hydrogen, and Z 2 is a heterocyclic group (eg, a nitrogen-containing heterocyclic group, eg, a nitrogen-containing spiroheterocyclic group, eg, 2-oxa-7-azaspiro[3.5]nonyl).

[0233] In some embodiments, the compound is a compound of formula (III-d). In some embodiments of formula (III-d), R 2a 、R 2b 、R2c and R 2d Each of is independently hydrogen, m is 1, n is 2, q is 1, 2, 3 or 4, p is 0, and X is S(O)2. In some embodiments of Formula (III-d), R 2a and R 2b Each of wherein is independently hydrogen, m is 1, n is 2, q is 1, 2, 3 or 4, p is 0, and X is S(O)2.

[0234] In some embodiments, the compound is a compound of formula (Ib), (Id), or (Ie). In some embodiments, the compound is a compound of formula (Ib), (Id), or (II). In some embodiments, the compound is a compound of formula (Ib), (Id), or (If). In some embodiments, the compound is a compound of formula (Ib), (Id), or (III).

[0235] In some embodiments, the compound of formula (I) is not a compound disclosed in WO2012 / 112982, WO2012 / 167223, WO2014 / 153126, WO2016 / 019391, WO 2017 / 075630, US2012-0213708, US 2016-0030359, or US 2016-0030360.

[0236] In some embodiments, the compound of formula (I) comprises a compound shown in Table 3, or a pharmaceutically acceptable salt thereof. In some embodiments, the exterior surface and / or one or more compartments within the devices described herein comprise a small molecule compound shown in Table 3, or a pharmaceutically acceptable salt thereof.

[0237] Table 3: Exemplary compounds of formula (I)

[0238]

[0239]

[0240]

[0241]

[0242]

[0243]

[0244] Conjugation of any of the compounds in Table 3 to a polymer (e.g., alginate) can be performed as described in Example 2 of WO 2019 / 195055 or any other suitable chemical reaction.

[0245] In some embodiments, the compound is a compound of Formula (I) (e.g., (Ia), (Ib), (Ic), (Id), (Ie), (If), (II), (II-a), (III), (III-a), (III-b), (III-c), (III-d), (III-e), (III-f), (III-g), (III-h), or (III-i)), or a pharmaceutically acceptable salt thereof, and is selected from:

[0246] 、 and , or a pharmaceutically acceptable salt thereof.

[0247] In some embodiments, the polysaccharide polymers or devices described herein (e.g., hydrogel capsules) comprise a compound or , or a pharmaceutically acceptable salt of either compound.

[0248] In some embodiments, the polysaccharide polymers or devices described herein (e.g., hydrogel capsules) comprise a compound

[0249] or , or a pharmaceutically acceptable salt of either compound.

[0250] In some embodiments, the polysaccharide polymers or devices described herein (e.g., hydrogel capsules) comprise a compound

[0251] 、 or , or a pharmaceutically acceptable salt of any one of these compounds.

[0252] In some embodiments, the compound of formula (I) (e.g., compound 101 in Table 3) is covalently attached to alginate (e.g., alginate with an approximate MW < 75 kDa, a G:M ratio ≥ 1.5) at a conjugation density of at least 2.0% and less than 9.0%, or 3.0% to 8.0%, 4.0%-7.0%, 5.0% to 7.0%, or 6.0%-7.0% or about 6.8%, as determined by combustion analysis for percentage nitrogen as described in WO 2020 / 069429.

[0253] Photoactive crosslinkers and photoinitiators

[0254] Described herein are polysaccharide polymers covalently bound to photoactive crosslinkers, and compositions and methods of use thereof. The photoactive crosslinker comprises a portion that is activated upon exposure to light. The light can include light of any wavelength from infrared to X-ray energy. In some embodiments, the light comprises ultraviolet light (e.g., between 360 nm and 400 nm, e.g., 370 nm to 390 nm, e.g., 380 nm to 400 nm, e.g., 390 nm to 400 nm). In some embodiments, the light comprises visible light (e.g., between 400 nm and 700 nm). The photoactive crosslinker often comprises at least one unsaturated functional group capable of free radical polymerization. In some embodiments, the photoactive crosslinker comprises an alkenyl group (e.g., C2-C 12 In some embodiments, the photoactive crosslinker comprises an alkynyl group (e.g., C2-C 12 Alkynyl, C2-C8 alkynyl). The moiety that can be activated after exposure to radiation includes aromatic groups, alkenyl, alkynyl and azido groups. Exemplary alkenyl compounds that can serve as photoactive crosslinkers include olefinic acids, such as acrylates, methacrylates, acrylamides and methacrylamides and their corresponding acid chlorides and anhydrides. In some embodiments, the photoactive crosslinker comprises an acrylate group. In some embodiments, the photoactive crosslinker comprises a methacrylate group. In some embodiments, the photoactive crosslinker comprises an acrylamide group. In some embodiments, the photoactive crosslinker comprises a methacrylamide group. Other exemplary alkenyl compounds include enols (e.g., 2-propylene-1-ol), alkenyl halides (such as allyl chloride, etc.), organometallic alkenyl compounds (such as vinyl magnesium bromide), and aryl compounds (e.g., styrene). Exemplary photoactive crosslinkers include acrylates, methacrylates, ethylene glycol dimethacrylate, divinylbenzene, 1,3-diisopropylbenzene and N,N'-methylenebisacrylamide. In one embodiment, the photoactive crosslinker is a bifunctional crosslinker, i.e., has two reactive functional groups. In one embodiment, the photoactive covalent crosslinker has an alkenyl and an amide functional group. In one embodiment, the photoactive crosslinker has an alkenyl and a carboxylate functional group. In one embodiment, the photoactive crosslinker has an alkenyl and an amide functional group.

[0255] In one embodiment, the photoactive crosslinker has formula (IV):

[0256] (IV), or a pharmaceutically acceptable salt or tautomer thereof, wherein X 1 Does not exist, is O, NR 33 or C(R 34a )(R 34b );R 30a 、R30b 、R 31 、R 32 、R 33 、R 34a and R 34b Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 Replace; and each R 7 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, oxo, hydroxy, cycloalkyl, or heterocyclyl.

[0257] In one embodiment, X 1 O, R 30a 、R 30b 、R 31 and R 32 Each of is hydrogen, and R 32 In one embodiment, X 1 O, R 30a 、R 30b 、R 31 and R 32 Each of is hydrogen, and R 32 is a heteroalkyl group (e.g., ethylamine, e.g., -CH2CH2NH2). In one embodiment, the photoactive crosslinker of formula (IV) is a methacrylate. In one embodiment, X 1 Does not exist; R 32 is a halo group (eg, a chloro group); and R 30a 、R 30b and R 31Each of is hydrogen. In one embodiment, the photoactive cross-linking agent of formula (IV) is acryloyl chloride.

[0258] In one embodiment, X 1 NR 33 (e.g., NH), and R 30a 、R 30b 、R 31 and R 32 Each of is hydrogen. In one embodiment, the photoactive cross-linker of formula (IV) is acrylamide.

[0259] In one embodiment, the photoactive crosslinker of formula (IV) has formula (IV-a):

[0260] (IV-a), or a pharmaceutically acceptable salt or tautomer thereof, wherein R 30a 、R 30b 、R 31 、R 32 and R 35 Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 Replace; and each R 7 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, oxo, hydroxy, cycloalkyl, or heterocyclyl.

[0261] In one embodiment, the photoactive crosslinker of formula (IV) has formula (IV-b):

[0262] (IV-b), or a pharmaceutically acceptable salt or tautomer thereof, wherein R30a 、R 30b 、R 31 、R 32 、R 36a and R 36b Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 Replace; each R 7 R is independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxy, cycloalkyl or heterocyclyl; 35 is hydrogen, alkyl, heteroalkyl, halo, cyano, nitro, amino, cycloalkyl, heterocyclyl, aryl, or heteroaryl; and n is 1, 2, 3, 4, 5, or 6.

[0263] In one embodiment, the photoactive crosslinker of formula (IV) has formula (IV-c):

[0264] (IV-c), or a pharmaceutically acceptable salt or tautomer thereof, wherein R 30a 、R 30b and R 31 Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SRE1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; R 32 is alkyl, alkenyl, alkynyl, heteroalkyl, –C(O)OR A1 ,–C(O)R B1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 Replace; and each R 7 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, oxo, hydroxy, cycloalkyl, or heterocyclyl.

[0265] In one embodiment, the photoactive crosslinker of formula (IV) has formula (IV-d):

[0266] (IV-d), or a pharmaceutically acceptable salt or tautomer thereof, wherein R 30a 、R 30b 、R 31 、R 32 、R 36a and R 36b Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; R 32 is alkyl, alkenyl, alkynyl, heteroalkyl, –C(O)OR A1 ,–C(O)R B1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、R C1 、R D1 and R E1are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 Replace; each R 7 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, oxo, hydroxy, cycloalkyl, or heterocyclyl; and n is 1, 2, 3, 4, 5, or 6.

[0267] In some embodiments, the photoactive crosslinker is a compound of Table 4.

[0268] Table 4. Exemplary photoactive crosslinkers of formula (IV)

[0269]

[0270]

[0271] Photoactive crosslinking agents can be used alone or, preferably, in the presence of a photoinitiator. As used herein, "photoinitiator" refers to a molecule that absorbs radiation (e.g., light, such as photons) and forms an active species in an excited state. Various free radical initiators readily recognized by those skilled in the art can be employed in the practice of the present invention. In one embodiment, the photoinitiator is an ultraviolet (UV) photoinitiator. Exemplary UV photoinitiators include lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP), camphorquinone, benzoin methyl ether, 1-hydroxy-cyclohexyl-phenyl-ketone (i.e., Irgacure 184), 2-hydroxy-2-methyl-1-phenyl-1-propanone (i.e., Darocur 1173), 2-hydroxy-1-[4-(2-hydroxyethoxy)phenyl]-2-methylpropan-1-one (i.e., Irgacure 2959), 2-benzyl-2-(dimethylamino)-1-(4-morpholin-4-ylphenyl)butan-1-one (i.e., Irgacure 369), 2-methyl-1-(4-methylthiophenyl)-2-morpholin-4-ylpropan-1-one (i.e., Irgacure 907), diphenyl(2,4,6-trimethylbenzoyl)phosphine oxide (e.g., Darocur In some embodiments, the photoinitiator is LAP. In some embodiments, the photoinitiator is camphorquinone. In some embodiments, the photoinitiator is benzoin methyl ether. In some embodiments, the photoinitiator is Irgacure 2959.

[0272] For visible light polymerization, a system of dye and co-catalyst can be used. Exemplary visible light photoinitiators include 2-(2,4,5,7-tetrabromo-3-hydroxy-6-oxoxanthene-9-yl)benzoic acid (i.e., Eosin Y), Erythrosine, riboflavin, Rose Bengal (rose Bengal), methylene blue and thionine. In some embodiments, the visible light photoinitiator is Eosin Y. In some embodiments, the visible light photoinitiator is Erythrosine. In some embodiments, the visible light photoinitiator is riboflavin. In some embodiments, the visible light photoinitiator is Rose Bengal. In some embodiments, the visible light photoinitiator is methylene blue. In some embodiments, the visible light photoinitiator is thionine. Optionally, a small amount of comonomer can be added to the cross-linking reaction to increase the polymerization rate. Examples of suitable comonomers include vinyl pyrrolidone, acrylamide, methacrylamide, acrylic acid, methacrylic acid, sodium acrylate, sodium methacrylate, hydroxyethyl acrylate, hydroxyethyl methacrylate (HEMA), ethylene glycol diacrylate, ethylene glycol dimethacrylate, pentaerythritol triacrylate, pentaerythritol trimethacrylate, trimethylolpropane triacrylate, trimethylolpropane trimethacrylate, tripropylene glycol diacrylate, tripropylene glycol dimethacrylate, glycerol acrylate, glycerol methacrylate, and the like.

[0273] In some embodiments, the photoinitiator is a thermally activated photoinitiator.

[0274] The photoactive crosslinker can be used in the presence of a single photoinitiator or a plurality of photoinitiators. The plurality of photoinitiators can include 2, 3, 4, 5, 6, 7, 8 or more photoinitiators.

[0275] In one embodiment, the covalently cross-linked portion is present on the polysaccharide polymer with a density of, for example, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20% or more as determined by LC-UV. In one embodiment, the covalently cross-linked portion is present on the polysaccharide polymer with a density of, for example, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20% or more as determined by LC-UV. In one embodiment, the covalently cross-linked portion is present on the polysaccharide polymer with a density of, for example, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20% or more as determined by LC-UV.

[0276] Modified polysaccharide can include photoactive cross-linking agent part.This photoactive cross-linking agent (for example, formula (IV)-(IV-d) compound) can be covalently bound to polysaccharide polymer, for example, alginate.This modified polysaccharide polymer (for example modified alginate polymer) may be able to cross-link with another polymer.In one embodiment, this polysaccharide polymer is modified through more than one type of photoactive cross-linking agent.

[0277] In one embodiment, the polysaccharide polymer is modified with a group capable of free radical polymerization. In one embodiment, the polysaccharide is modified with a compound of any one of formula (IV), (IV-a), (IV-b), (IV-c) or (IV-d). In one embodiment, the modified polysaccharide is modified with a compound selected from Table 4. In one embodiment, the polysaccharide is modified with a compound selected from Table 4. or In one embodiment, the polysaccharide polymer is modified Modified.

[0278] In one embodiment, the modified polysaccharide is a compound of formula (V):

[0279] (V), or a pharmaceutically acceptable salt or tautomer thereof, wherein each of T and U is independently C(R 40 )(R 41 ), O or N(R 42 );R 38a 、R 38b 、R 39a 、R 39b 、R 40 、R 41 and R 42 Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; R 32 and R 35 Each of is hydrogen, alkyl, heteroalkyl, halo, cyano, nitro, amino, cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、RC1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 Replace; each R 7 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxyl, cycloalkyl or heterocyclic; and the photoactive crosslinker has a structure of formula (IV), (IV-a), (IV-b), (IV-c) or (IV-d).

[0280] In one embodiment, the modified polysaccharide polymer of formula (V) has formula (Va):

[0281] (Va), or a pharmaceutically acceptable salt or tautomer thereof, wherein each of T and U is independently C(R 40 )(R 41 ), O or N(R 42 );R 30a 、R 30b 、R 31 、R 32 、R 38a 、R 38b 、R 39a 、R 39b 、R 40 、R 41 and R 42 Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7Replace; and each R 7 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, oxo, hydroxy, cycloalkyl, or heterocyclyl.

[0282] In one embodiment, the modified polysaccharide polymer of formula (V) has formula (Vb):

[0283] (Vb), or a pharmaceutically acceptable salt or tautomer thereof, wherein each of U and T is independently C(R 40 )(R 41 ), O or N(R 42 );R 30a 、R 30b 、R 31 、R 35 、R 38a 、R 38b 、R 39a 、R 39b 、R 40 、R 41 、R 42 、R 43a and R 43b Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 Replace; each R 7 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, oxo, hydroxy, cycloalkyl, or heterocyclyl; and n is 1, 2, 3, 4, 5, or 6.

[0284] In one embodiment, the modified polysaccharide polymer of formula (V) has formula (Vc):

[0285] (Vc), or a pharmaceutically acceptable salt or tautomer thereof, wherein U is C(R 40 )(R 41 ), O or N(R 42 );R 30a 、R 30b 、R 31 、R 35 、R 38a 、R 38b 、R 39a 、R 39b 、R 40 、R 41 、R 42 、R 43a 、R 43b and R 44 Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 Replace; and each R 7 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, oxo, hydroxy, cycloalkyl, or heterocyclyl; and n is 1, 2, 3, 4, 5, or 6.

[0286] In one embodiment, the modified polysaccharide polymer of formula (V) has formula (Vd):

[0287] (Vd), or a pharmaceutically acceptable salt or tautomer thereof, wherein U is C(R 40 )(R 41 ), O or N(R 42 );R 30a 、R30b 、R 31 、R 38a 、R 38b 、R 39a 、R 39b 、R 40 、R 41 、R 42 、R 43a and R 43b Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 Replace; each R 7 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, oxo, hydroxy, cycloalkyl, or heterocyclyl; and n is 1, 2, 3, 4, 5, or 6.

[0288] In one embodiment, the modified polysaccharide polymer comprises formula (VI):

[0289] (VI), or a pharmaceutically acceptable salt or tautomer thereof, wherein W, T 1 、T 2 、U 1 and U 2 Each of which is independently C(R 40 )(R 41 ), O or N(R 42 );R 38a 、R 38b 、R 38c 、R 38d R 39a 、R 39b 、R 39a 、R 39b、R 40 、R 41 and R 42 Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 Replace; and each R 7 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxyl, cycloalkyl or heterocyclyl; p is an integer from 1 to 100; the defiberizing compound has a structure of Formula (I), (Ia), (Ib), (Ibi), (Ib-ii), (Ic), (Id), (Ie), (If), (II), (II-a), (III), (III-a), (III-b), (III-c), (III-d), (III-e), (III-f), (III-g), (III-h) or (III-i); and the photoactive crosslinker has a structure of Formula (IV), (IV-a), (IV-b), (IV-c), (IV-d) or (IV-e).

[0290] In one embodiment, the modified polysaccharide polymer comprises formula (VI-ai):

[0291] The structure of (VI-ai), or a pharmaceutically acceptable salt or tautomer thereof, wherein W, T 1 、U 1 and U 2 Each of which is independently C(R 40 )(R 41 ), O or N(R 42 );R 38a 、R 38b、R 38c 、R 38d 、R 39a 、R 39b 、R 39c 、R 39d 、R 40 、R 41 and R 42 Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 Replace; and each R 7 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, oxo, hydroxy, cycloalkyl, or heterocyclyl; the variable M 1 、Z 1 、R 2a 、R 2b 、R 2c 、R 2d , X, R C , m and n are as defined in formula (Ib); p is an integer from 1 to 100; and the photoactive crosslinker has a structure of formula (IV), (IV-a), (IV-b), (IV-c), (IV-d) or (IV-e).

[0292] In one embodiment, the modified polysaccharide polymer comprises formula (VI-a-ii):

[0293] The structure of (VI-a-ii), or a pharmaceutically acceptable salt or tautomer thereof, wherein W, T 1 、U 1 and U 2 Each of which is independently C(R 40 )(R41 ), O or N(R 42 );R 38a 、R 38b 、R 38c 、R 38d 、R 39a 、R 39b 、R 39c 、R 39d 、R 40 、R 41 and R 42 Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 Replace; and each R 7 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxy, cycloalkyl or heterocyclyl; variable Z 1 、R 2a 、R 2b 、R 2c 、R 2d 、R C , m, n and q are as defined in formula (III-f); p is an integer from 1 to 100; and the photoactive crosslinker has a structure of formula (IV), (IV-a), (IV-b), (IV-c), (IV-d) or (IV-e).

[0294] In one embodiment, the modified polysaccharide polymer comprises formula (VI-bi):

[0295] The structure of (VI-bi), or a pharmaceutically acceptable salt or tautomer thereof, wherein W, X 1 、T 2 、U1 and U 2 Each of which is independently C(R 40 )(R 41 ), O or N(R 42 );R 30a 、R 30b 、R 31 、R 32 、R 38a 、R 38b 、R 38c 、R 38d 、R 39a 、R 39b 、R 39c 、R 39d 、R 40 、R 41 、R 42 and R 44 Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 Replace; and each R 7 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxy, cycloalkyl or heterocyclyl; and the defiberizing compound has the structure of Formula (I), (Ia), (Ib), (Ibi), (Ib-ii), (Ic), (Id), (Ie), (If), (II), (II-a), (III), (III-a), (III-b), (III-c), (III-d), (III-e), (III-f), (III-g), (III-h) or (III-i).

[0296] In one embodiment, the polysaccharide polymer comprises formula (VI-ci):

[0297] (VI-ci), or a pharmaceutically acceptable salt or tautomer thereof, wherein X 1 、U 1 、U 2 Each of W is independently C(R 40 )(R 41 ), O or N(R 42 );R 30a 、R 30b 、R 31 、R 32 、R 38a 、R 38b 、R 38c 、R 38d 、R 39a 、R 39b 、R 39c 、R 39d 、R 40 、R 41 、R 42 and R 44 Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 Replace; and each R 7 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, oxo, hydroxy, cycloalkyl, or heterocyclyl; the variable M 1 、Z 1 、R 2a 、R 2b 、R 2c 、R 2d, X, R C , m and n are as defined in formula (Ib); and p is an integer from 1 to 100.

[0298] In one embodiment, the polysaccharide polymer comprises formula (VI-c-ii):

[0299] The structure of (VI-c-ii), or a pharmaceutically acceptable salt or tautomer thereof, wherein W, X 1 、Y 1 and Y 2 Each of which is independently C(R 40 )(R 41 ), O or N(R 42 );R 30a 、R 30b 、R 31 、R 32 、R 38a 、R 38b 、R 38c 、R 38d 、R 39a 、R 39b 、R 39c 、R 39d 、R 40 、R 41 、R 42 and R 44 Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 Replace; and each R 7 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxy, cycloalkyl or heterocyclyl; variable Z 1、R 2a 、R 2b 、R 2c 、R 2d 、R C , m, n and q are as defined in formula (III-f); and p is an integer from 1 to 100.

[0300] In one embodiment, the modified polysaccharide polymer of formula (VI) is a compound selected from Table 5:

[0301] Table 5. Exemplary compounds of formula (VI) or pharmaceutically acceptable salts thereof.

[0302]

[0303]

[0304]

[0305]

[0306]

[0307] In one embodiment, the modified polysaccharide polymer is 、 、 、 or .

[0308] Polysaccharide polymers described herein can be modified on any suitable functional group (e.g., carboxyl or hydroxyl). In one embodiment, the polysaccharide polymer is modified on a single type of functional group. In one embodiment, the polysaccharide polymer is modified on more than one type of functional group. In one embodiment, the polysaccharide polymer is modified on a carboxyl group. In one embodiment, the polysaccharide polymer is modified on a carboxyl group and a hydroxyl group. In one embodiment, the polysaccharide polymers described herein can be modified by compounds of formula (I) and / or compounds of formula (IV) on one or more functional groups.

[0309] In one embodiment, the degree of modification (i.e., the percentage of functional groups of the polymer modified with a photoactive crosslinker) is about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99%. In one embodiment, the degree of modification (i.e., the percentage of functional groups of the polymer modified with a photoactive crosslinker) is greater than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99%. In one embodiment, the degree of modification (i.e., the percentage of functional groups of the polymer that are modified with the photoactive cross-linker) is less than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%.

[0310] In some embodiments, for example, when the polysaccharide polymers described herein are used to prepare hydrogel capsules having dual crosslinking, the polymers retain sufficient unreacted carboxylic acid groups to allow ionic crosslinking. In some embodiments, the polysaccharide polymers described herein do not contain more than 10% modified carboxylic acid groups. In some embodiments, the polysaccharide polymers described herein do not contain more than 5% modified carboxylic acid groups. In some embodiments, the polysaccharide polymers described herein do not contain more than 5%, 6%, 7%, 8%, 9%, or 10% modified carboxylic acid groups.

[0311] In one embodiment, the polysaccharide polymers described herein are modified with one, two, three or more unique compounds. In one embodiment, the polysaccharide polymers described herein are modified with a photoactive cross-linking agent (e.g., a compound of formula (IV), a compound of formula (I) and a cell adhesion molecule (e.g., any of the cell binding peptides described herein, e.g., RGD or RGDSP)). In one embodiment, the polysaccharide polymers described herein are modified with a photoactive cross-linking agent. In one embodiment, the polysaccharide polymers described herein are modified with a compound of formula (I). In one embodiment, the polysaccharide polymers described herein are modified with a cell adhesion molecule. In one embodiment, the polysaccharide polymers described herein are modified with both a photoactive cross-linking agent and a cell adhesion molecule. In a preferred embodiment, the polysaccharide polymers described herein are modified with both a photoactive cross-linking agent and a compound of formula (I). In one embodiment, the polysaccharide polymers described herein are modified with both a photoactive cross-linking agent (e.g., compound 201) and a compound of formula (I) (e.g., a compound from Table 3).

[0312] In one embodiment, the polysaccharide polymers described herein are and modified with compounds from Table 3. In one embodiment, the polysaccharide polymers described herein are and modified with compounds from Table 3. In one embodiment, the polysaccharide polymers described herein are and modified with compounds from Table 3. In one embodiment, the polysaccharide polymers described herein are and modified with compounds from Table 3. In one embodiment, the polysaccharide polymers described herein are and modified with compounds from Table 3. In one embodiment, the polysaccharide polymers described herein are and modified with compounds from Table 3. In one embodiment, the polysaccharide polymers described herein are and a compound from Table 3. In one embodiment, the polysaccharide polymers described herein are modified with and a compound from Table 3.

[0313] In one embodiment, the polysaccharide polymers described herein are and compounds In one embodiment, the polysaccharide polymers described herein are Modified alginate .

[0314] In one embodiment, polysaccharide polymer described herein is modified through cell adhesion molecules (for example, cell binding peptides) and formula (I) compounds. In some embodiments, the polysaccharide polymer modified through one or both of photoactive cross-linking agents and optional formula (I) compounds (for example, from Table 3) and cell binding peptides is alginate, such as VLVG alginate or SLG-100 alginate, such as low viscosity alginate, such as high G:M than alginate. In some embodiments, polysaccharide polymer described herein is modified through formula (I) compounds and formula (IV) compounds. In some embodiments, polysaccharide polymer described herein is modified through formula (I) compounds, formula (IV) compounds, and does not include heparin. In some embodiments, polysaccharide polymer described herein is modified through formula (I) compounds, formula (IV) compounds, and does not include anti-CD3 antibodies. In some embodiments, polysaccharide polymer described herein is modified through formula (I) compounds, formula (IV) compounds, and does not include anti-CD28 antibodies. In some embodiments, the polysaccharide polymers described herein are modified with a compound of formula (I) or a compound of formula (IV) and do not contain a major histocompatibility peptide. In some embodiments, the polysaccharide polymers described herein are modified with a compound of formula (I) or a compound of formula (IV) and do not contain heparin, anti-CD3 antibodies, anti-CD28 antibodies, or major histocompatibility peptides.

[0315] The hydroxyl groups of the polysaccharide (eg, alginate) may be modified to improve one or more physical properties.The polysaccharide polymers of the present invention may be sulfonated or may be non-sulfonated (ie, should not contain sulfate groups).

[0316] Characteristics of hydrogels and hydrogel capsules

[0317] The present disclosure further features hydrogels and hydrogel capsules comprising the polysaccharide polymers described herein. The hydrogels and hydrogel capsules can be crosslinked (i.e., covalently crosslinked) by photoactive crosslinking groups and / or by, for example, the presence of divalent cations (e.g., Ba 2- ). In one embodiment, the hydrogels or hydrogel capsules described herein are produced by photoactivated crosslinking. In one embodiment, the hydrogels or hydrogel capsules described herein are produced by photoactivated crosslinking and ionic crosslinking (also referred to herein as "double crosslinking"). Those skilled in the art will recognize that other methods of initiating polymerization are also possible, including application of heat, ultrasound, and gamma radiation in the presence of an appropriate initiator.

[0318] The polysaccharide polymers described herein that contain photoactive crosslinker moieties may be capable of undergoing further polymerization, for example, reacting with compatible functional groups on the same or different polymers. In one embodiment, a polymer modified to contain thiol groups and a second polymer modified to contain olefin groups allow for the formation of a crosslinked polymer by reacting the first polymer and the second polymer. In some embodiments, the hydrogel capsules are formed by covalent crosslinking of unsaturated functional groups through a chain growth polymerization process. In one embodiment, the hydrogel capsules are formed by covalent crosslinking of unsaturated functional groups on the first polymer with unsaturated functional groups on the second polymer. In a preferred embodiment, the hydrogel capsules are formed by covalent crosslinking of olefin functional groups on the first polymer with olefin functional groups on the second polymer. In other embodiments, the hydrogel capsules are formed by covalent crosslinking of unsaturated functional groups through a step-growth polymerization process. In one embodiment, the step-growth polymerization process comprises a reaction between one or more unsaturated functional groups (e.g., olefinic groups) of one polysaccharide chain and a thiolated functional group of another polymer chain.

[0319] The hydrogel capsules described herein are formed by cross-linking one or more types of polysaccharide polymers. In one embodiment, the hydrogel capsule comprises only polysaccharide polymers. In one embodiment, the hydrogel capsule comprises the same type of polysaccharide polymer, such as an alginate polymer. In one embodiment, the hydrogel capsule is formed by the polymerization of two identical polysaccharides. In one embodiment, the hydrogel capsule is formed by the polymerization of two different polysaccharides. In one embodiment, the hydrogel capsule comprises a plurality of polymers, such as a plurality of polysaccharide polymers. In one embodiment, the hydrogel capsule comprises a polysaccharide polymer and a non-polysaccharide polymer.

[0320] The hydrogel capsules described herein may be homogeneous, i.e., may not contain non-polysaccharide polymers. In one embodiment, the hydrogel capsules described herein do not contain a polymer selected from polyacrylamide, poly(vinyl alcohol), poly(ethylene oxide), polyethylene glycol (PEG), and polyphosphazene. In one embodiment, the hydrogel capsule does not contain poly(vinyl alcohol). In one embodiment, the hydrogel capsule does not contain poly(ethylene oxide). In one embodiment, the hydrogel capsule does not contain polyethylene glycol (PEG). In one embodiment, the hydrogel capsule does not contain polyphosphazene. In one embodiment of the present invention, the hydrogel capsule is a two-compartment hydrogel capsule. In a preferred embodiment of the present disclosure, the hydrogel capsule consists of an inner compartment and an outer compartment. In one embodiment, the two compartments are formed from the same type of modified polysaccharide. In one embodiment, the two compartments are formed from different types of modified polysaccharides. In one embodiment, the inner compartment is formed from a polysaccharide of formula (Vb) and the outer compartment is formed from a polysaccharide of formula (Vb). In one embodiment, the inner compartment is formed from a polysaccharide of formula (Vd) and the outer compartment is formed from a polysaccharide of formula (Vd). In one embodiment, the inner compartment is formed by the polysaccharide of formula (VI-ci) and the outer compartment is formed by the polysaccharide of formula (VI-ci).

[0321] In some embodiments, the inner compartment is formed by an unmodified polysaccharide, and the outer compartment is formed by a polysaccharide modified with a compound of formula (Vb), (Vd) or (VI-ci). In some embodiments, the outer compartment is formed by an unmodified polysaccharide, and the inner compartment is formed by a polysaccharide modified with a compound of formula (vb), (Vd) or (VI-ci). In some embodiments, the inner compartment is formed by a polysaccharide modified with a cell-binding substance (e.g., a compound of Table 1), and the outer compartment is formed by a polysaccharide modified with a compound of formula (Vb), (Vd) or (VI-ci). In some embodiments, the inner compartment is formed by a polysaccharide modified with a compound of Table 1, and the outer compartment is formed by a polysaccharide modified with a compound of formula (Vb), (Vd) or (VI-ci).

[0322] The present disclosure is characterized in that a dual cross-linked polysaccharide for encapsulating mammalian cells. In one embodiment, the dual cross-linked polysaccharide polymer may further include at least one cell binding peptide (CBP) (as defined herein). When a device (e.g., a hydrogel capsule) comprising the dual cross-linked polysaccharide polymer is implanted in a subject (e.g., a human or other mammalian subject), the cell can express a therapeutic agent. In addition, the device includes at least one means for alleviating FBR (as defined herein). In one embodiment, the means for alleviating FBR includes a defibrotic compound as defined herein. In one embodiment, the defibrotic compound is covalently bound to the dual cross-linked polysaccharide polymer.

[0323] In some embodiments, the device (e.g., hydrogel capsule) further comprises an unmodified polysaccharide polymer. In some embodiments, the unmodified polysaccharide polymer is an unmodified alginate. In some embodiments, the alginate is a high guluronic acid (G) alginate and comprises greater than about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or more guluronic acid (G). In some embodiments, the alginate is a high mannuronic acid (M) alginate and comprises greater than about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or more mannuronic acid (M). In some embodiments, the M:G ratio is about 1. In some embodiments, the M:G ratio is less than 1. In some embodiments, the M:G ratio is greater than 1. In one embodiment, the unmodified alginate has a molecular weight of 150 kDa – 250 kDa and a G:M ratio of ≥ 1.5.

[0324] In some embodiments, the equipment (such as hydrogel capsule) includes modified polysaccharide polymer, and it is defibration polymer, for example, includes the defibration alginate of the alginate chemically modified by formula (I) compound.The alginate in defibration polymer can be identical or different with any unmodified alginate present in equipment.In one embodiment, the density (for example, conjugated amount) of defibration alginate formula (I) compound is between about 4.0% and about 8.0% nitrogen, between about 5.0% and about 7.0% nitrogen or between about 6.0% and about 7.0% nitrogen (for example, as determined by the combustion analysis of nitrogen percentage).In one embodiment, the amount of compound 101 produces about 0.5% to 2%, 2% to 4% N, about 4% to 6% N, about 6% to 8%, or about 8% to 10% N %N increase (compared with unmodified alginate), wherein %N is determined by combustion analysis and corresponds to the amount of the compound 101 in modified alginate.

[0325] Hydrogel capsules described herein can be porous or non-porous. The hole in the polysaccharide hydrogel capsule (for example, the polysaccharide hydrogel capsule formed by alginate hydrogel) serves as the selective permeability membrane of small proteins and molecules, while preventing larger unwanted molecules (such as immunoglobulins) from entering the cells through encapsulation. In a preferred embodiment, hydrogel described herein and hydrogel capsules are porous. In some embodiments, the mean pore size is about 1 nm, 2 nm, 3 nm, 4 nm, 5 nm, 6 nm, 7 nm, 8 nm, 9 nm, 10 nm, 11 nm, 12 nm, 13 nm, 14 nm, 15 nm, 16 nm, 17 nm, 18 nm, 19 nm or 20 nm. In some embodiments, the mean pore size is greater than about 1 nm, 2 nm, 3 nm, 4 nm, 5 nm, 6 nm, 7 nm, 8 nm, 9 nm, 10 nm, 11 nm, 12 nm, 13 nm, 14 nm, 15 nm, 16 nm, 17 nm, 18 nm, 19 nm or 20 nm. In some embodiments, the average pore size is less than about 1 nm, 2 nm, 3 nm, 4 nm, 5 nm, 6 nm, 7 nm, 8 nm, 9 nm, 10 nm, 11 nm, 12 nm, 13 nm, 14 nm, 15 nm, 16 nm, 17 nm, 18 nm, 19 nm, or 20 nm.

[0326] In some embodiments, the average pore size of the first compartment and the second compartment of the particle (e.g., a hydrogel capsule) is substantially the same. In some embodiments, the average pore size of the first compartment and the second compartment of the particle differs by about 1.5%, 2%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% or more. In some embodiments, the average pore size of the particle (e.g., the average pore size of the first compartment and / or the average pore size of the second compartment) depends on various factors, such as the one or more materials in each compartment, the presence and density of the photoactive cross-linking agent, the presence and density of the compound of formula (I).

[0327] Hydrogel capsules described herein should not have the hole that is enough to allow cell (for example, immune cell, such as dendritic cell) to move through the diameter of hydrogel.In some embodiments, the diameter of hole is small enough to prevent antibody from moving through hydrogel.In some embodiments, hydrogel capsules described herein do not have the pore size greater than 75 μ m.In some embodiments, hydrogel capsules described herein do not have the pore size greater than 1 μ m, 2 μ m, 3 μ m, 4 μ m, 5 μ m, 6 μ m, 7 μ m, 8 μ m, 9 μ m, 10 μ m, 15 μ m, 20 μ m, 25 μ m, 30 μ m, 35 μ m, 40 μ m, 45 μ m, 50 μ m, 55 μ m, 60 μ m, 65 μ m, 70 μ m or 75 μ m.

[0328] The physical properties of the hydrogel capsules described herein (e.g., as described in the Examples) control the release of encapsulated molecules and / or limit the absorption or permeation of unwanted molecules outside the capsule (e.g., as determined by dextran permeability assay). In some embodiments, the average molecular weight permeability is about 1 kDa to about 150 kDa. In some embodiments, the average molecular weight permeability is about 1 kDa, 5 kDa, 10 kDa, 15 kDa, 20 kDa, 25 kDa, 30 kDa, 35 kDa, 40 kDa, 45 kDa, 50 kDa, 55 kDa, 60 kDa, 65 kDa, 70 kDa, 75 kDa, 80 kDa, 85 kDa, 90 kDa, 95 kDa, 100 kDa, 105 kDa, 110 kDa, 115 kDa, 120 kDa, 125 kDa, 130 kDa, 135 kDa, 140kDa, 145 kDa, 150 kDa, 155 kDa, 160 kDa, 165 kDa, 170 kDa, 175 kDa, 180 kDa, 185 kDa, 190 kDa, 195 kDa, or 200 kDa. In some embodiments, the average molecular weight permeability is greater than about 1 kDa, 5 kDa, 10 kDa, 15 kDa, 20 kDa, 25 kDa, 30 kDa, 35 kDa, 40 kDa, 45 kDa, 50 kDa, 55 kDa, 60 kDa, 65 kDa, 70 kDa, 75 kDa, 80 kDa, 85 kDa, 90 kDa, 95 kDa, 100 kDa, 105 kDa, 110 kDa, 115 kDa, 120 kDa, 125 kDa, 130 kDa, 135 kDa, 140 kDa, 145 kDa, 150 kDa, 155 kDa, 160 kDa, 165 kDa, 170 kDa, 175 kDa, 180 kDa, 185 kDa, 190 kDa, 195 kDa, or 200 kDa.In some embodiments, the average molecular weight permeability is less than about 1 kDa, 5 kDa, 10 kDa, 15 kDa, 20 kDa, 25 kDa, 30 kDa, 35 kDa, 40 kDa, 45 kDa, 50 kDa, 55 kDa, 60 kDa, 65 kDa, 70 kDa, 75 kDa, 80 kDa, 85 kDa, 90 kDa, 95 kDa, 100 kDa, 105 kDa, 110 kDa, 115 kDa, 120 kDa, 125 kDa, 130 kDa, 135 kDa, 140 kDa, 145 kDa, 150 kDa, 155 kDa, 160 kDa, 165 kDa, 170 kDa, 175 kDa, 180 kDa, 185 kDa, 190 kDa, 195 kDa, or 200 kDa.

[0329] In some embodiments, the hydrogel capsules described herein can be characterized by their absolute burst strength (e.g., crush strength) as determined using a texture analyzer. In some embodiments, the absolute burst strength is between 50 and 800 g. In some embodiments, the hydrogel capsules described herein have a mass of about 50 g, 60 g, 70 g, 80 g, 90 g, 100 g, 110 g, 120 g, 130 g, 140 g, 150 g, 160 g, 170 g, 180 g, 190 g, 200 g, 210 g, 220 g, 230 g, 240 g, 250 g, 260 g, 270 g, 280 g, 290 g, 300 g, 310 g, 320 g, 330 g, 340 g, 350 g, 360 g, 370 g, 380 g, 390 g, 400 g, 410 g, 420 g, 430 g, 440 g, 450 g, 460 g, 470 g, 480 g, 490 g, 500 g, 510 g, 520 g, 700 g, 780 g, 790 g, or an absolute strength of 800 g.In some embodiments, the hydrogel capsules described herein have a mass greater than about 50 g, 60 g, 70 g, 80 g, 90 g, 100 g, 110 g, 120 g, 130 g, 140 g, 150 g, 160 g, 170 g, 180 g, 190 g, 200 g, 210 g, 220 g, 230 g, 240 g, 250 g, 260 g, 270 g, 280 g, 290 g, 300 g, 310 g, 320 g, 330 g, 340 g, 350 g, 360 g, 370 g, 380 g, 390 g, 400 g, 410 g, 420 g, 430 g, 440 g, 450 g, 460 g, 470 g, 480 g, 490 g, 500 g, 510 g, 520 g, 700 g, 780 g, 790 g, or an absolute strength of 800 g. In some embodiments, the hydrogel capsules described herein have a mass of less than about 50 g, 60 g, 70 g, 80 g, 90 g, 100 g, 110 g, 120 g, 130 g, 140 g, 150 g, 160 g, 170 g, 180 g, 190 g, 200 g, 210 g, 220 g, 230 g, 240 g, 250 g, 260 g, 270 g, 280 g, 290 g, 300 g, 310 g, 320 g, 330 g, 340 g, 350 g, 360 g, 370 g, 380 g, 390 g, 400 g, 410 g, 420 g, 430 g, 440 g, 450 g, 460 g, 470 g, 480 g, 490 g, 500 g, 510 g, 520 g, 700 g, 780 g, 790 g, or an absolute strength of 800 g.

[0330] The present disclosure features particles (e.g., hydrogel capsules) comprising a first compartment, a second compartment, a photoactive cross-linking moiety as described herein (e.g., a compound of Formula (IV, V, or VI)), and optionally a compound of Formula (I), such as described herein. The photoactive cross-linking moiety is covalently bonded to a polysaccharide polymer present in the first and / or second compartment. The particles (e.g., hydrogel capsules) can be spherical or have any other shape. The particles (e.g., hydrogel capsules) can comprise materials such as metals, metal alloys, ceramics, polymers, fibers, inert materials, and combinations thereof. The particles (e.g., hydrogel capsules) can be made entirely of one type of material, or can contain a number of other materials within the second (outer) compartment and the first (inner) compartment.

[0331] In some embodiments, the first compartment is modified with a compound of formula (I). In some embodiments, the second compartment is modified with a compound of formula (I). In some embodiments, both the first compartment and the second compartment are independently modified with a compound of formula (I).

[0332] In some embodiments, the particles (e.g., hydrogel capsules) have a maximum linear dimension (LLD) (e.g., average diameter) or size greater than 1 millimeter (mm), preferably 1.5 mm or greater. In some embodiments, the diameter or size of the particles (e.g., hydrogel capsules) can be as large as 10 mm. For example, the particles described herein (e.g., hydrogel capsules) have a size range of 0.5 mm to 10 mm, 1 mm to 10 mm, 1 mm to 8 mm, 1 mm to 6 mm, 1 mm to 5 mm, 1 mm to 4 mm, 1 mm to 3 mm, 1 mm to 2 mm, 1 mm to 1.5 mm, 1.5 mm to 8 mm, 1.5 mm to 6 mm, 1.5 mm to 5 mm, 1.5 mm to 4 mm, 1.5 mm to 3 mm, 1.5 mm to 2 mm, 2 mm to 8 mm, 2 mm to 7 mm, 2 mm to 6 mm, 2 mm to 5 mm, 2 mm to 4 mm, 2 mm to 3 mm, 2.5 mm to 8 mm, 2.5 mm to 7 mm, 2.5 mm to 6 mm, 2.5 mm to 5 mm, 2.5 mm to 4 mm, 2.5 mm to 3 mm, 3 mm to 8 mm, 3 mm to 7 mm, 3 mm to 6 mm, 3 mm to 5 mm, 3 mm to 4 mm, 3.5 mm to 8 In some embodiments, the particles (e.g., hydrogel capsules) have an average diameter or size of between about 1 mm and about 8 mm. In some embodiments, the particles (e.g., hydrogel capsules) have an average diameter or size of between about 1 mm and about 4 mm. In some embodiments, the particles (e.g., hydrogel capsules) have an average diameter or size of between about 1 mm and about 4 mm. In some embodiments, the particles (e.g., hydrogel capsules) have an average diameter or size between about 1 mm and about 2 mm. In some embodiments, the particles (e.g., hydrogel capsules) have an average diameter or size between about 1.5 mm and about 2 mm.

[0333] In some embodiments, the particles (e.g., hydrogel capsules) have a maximum linear dimension (LLD) (e.g., mean diameter) or size of 1 millimeter (mm) or less. In some embodiments, the particles (e.g., hydrogel capsules) have a size range of 0.3 mm to 1 mm, 0.4 mm to 1 mm, 0.5 mm to 1 mm, 0.6 mm to 1 mm, 0.7 mm to 1 mm, 0.8 mm to 1 mm, or 0.9 mm to 1 mm.

[0334] In some embodiments, the second (outer) compartment completely surrounds the first (inner) compartment, and the inner boundary of the second compartment forms an interface with the outer boundary of the first compartment. In such embodiments, the thickness of the second (outer) compartment means the average distance between the outer boundary of the second compartment and the interface between the two compartments. In some embodiments, the thickness of the outer compartment is greater than about 10 nanometers (nm), preferably 100 nm or larger, and can be as large as 1 mm. For example, the thickness of the outer compartment in the particles described herein can be 10 nanometers to 1 millimeter, 100 nanometers to 1 millimeter, 500 nanometers to 1 millimeter, 1 micron (µm) to 1 millimeter, 1 µm to 1 mm, 1 µm to 500 µm, 1 µm to 250 µm, 1 µm to 1 mm, 5 µm to 500 µm, 5 µm to 250 µm, 10 µm to 1mm, 10 µm to 500 µm, or 10 µm to 250 µm. In some embodiments, the outer compartment has a thickness of 100 nanometers to 1 millimeter, between 1 μm and 1 mm, between 1 μm and 500 μm, or between 5 μm and 1 mm.

[0335] In some embodiments, the particles (e.g., hydrogel capsules) include at least one hole or opening, such as to allow free flow of material. In some embodiments, the average pore size of the particles is between about 0.1 μm and about 10 μm. For example, the average pore size can be between 0.1 μm and 10 μm, 0.1 μm and 5 μm, 0.1 μm and 2 μm, 0.15 μm and 10 μm, 0.15 μm and 5 μm, 0.15 μm and 2 μm, 0.2 μm and 10 μm, 0.2 μm and 5 μm, 0.25 μm and 10 μm, 0.25 μm and 5 μm, 0.5 μm and 10 μm, 0.75 μm and 10 μm, 1 μm and 10 μm, 1 μm and 5 μm, 1 μm and 2 μm, 2 μm and 10 μm, 2 μm and 5 μm, or 5 μm and 10 μm. In some embodiments, the average pore size of the particle is between about 0.1 μm and 10 μm. In some embodiments, the average pore size of the particle is between about 0.1 μm and 5 μm. In some embodiments, the average pore size of the particle is between about 0.1 μm and 1 μm. In some embodiments, the average pore size of the first compartment and the second compartment of the particle is substantially the same. In some embodiments, the average pore size of the first compartment and the second compartment of the particle differs by about 1.5%, 2%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% or more. In some embodiments, the average pore size of the particle (for example, the average pore size of the first compartment and / or the average pore size of the second compartment) depends on multiple factors, such as the presence and density of one or more materials and formula (I) compound in each compartment.

[0336] In some embodiments, particle (for example, hydrogel capsule) comprises polysaccharide, and this polysaccharide is alginate.Alginate is the polysaccharide being made up of β-D-mannuronic acid (M) and α-L-guluronic acid (G).In some embodiments, alginate is high guluronic acid (G) alginate, and comprises and is greater than about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or more guluronic acid (G).In some embodiments, alginate is high mannuronic acid (M) alginate, and comprises and is greater than about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or more mannuronic acid (M).In some embodiments, M:G ratio is about 1.In some embodiments, M:G ratio is less than 1.In some embodiments, M:G ratio is greater than 1. In the alginate-containing particles, the amount of alginate (e.g., the actual weight of alginate as a % by weight of the particles) may be at least 5%, such as at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99% or more, e.g., w / w; less than 20%, such as less than 20%, 15%, 10%, 5%, 1%, 0.5%, 0.1% or less.

[0337] In some embodiments, the first compartment and the second compartment both comprise the same polymer. In some embodiments, the first compartment and the second compartment comprise different polymers. In some embodiments, the first compartment comprises alginate. In some embodiments, the second compartment comprises alginate. In some embodiments, the first compartment and the second compartment both comprise alginate. In some embodiments, the alginate in the first compartment is different from the alginate in the second compartment. In some embodiments, the first compartment comprises alginate and the second compartment comprises different polymers (e.g., polysaccharides, such as hyaluronate or chitosan). In some embodiments, the second compartment comprises alginate and the first compartment comprises different polymers (e.g., polysaccharides, such as hyaluronate or chitosan).

[0338] The first compartment and the second compartment can both include a single component (e.g., a polymer) or more than one component (e.g., a blend of polymers). In some embodiments, the first compartment contains only alginate (e.g., a chemically modified alginate, or a blend of unmodified alginate and a chemically modified alginate). In some embodiments, the second compartment contains only alginate (e.g., a chemically modified alginate, or a blend of unmodified alginate and a chemically modified alginate). In some embodiments, the first and second compartments each independently contain only alginate (e.g., a chemically modified alginate, or a blend of unmodified alginate and a chemically modified alginate).

[0339] In some embodiments, the first and second compartments comprise a blend of polymers (i.e., a mixture of polymers). In some embodiments, the first (inner) compartment comprises a blend of polymers. In some embodiments, the second (outer) compartment comprises a blend of polymers. In some embodiments, the first and second compartments comprise a blend of identical polymers. In some embodiments, the first and second compartments comprise a blend of different polymers. In some embodiments, at least one polymer in the blend comprising the outer compartment is covalently modified with a photoactive crosslinker as described herein (e.g., a compound of formula (IV), (V) or (VI)). In some embodiments, at least one polymer in the blend comprising the second (outer) compartment is covalently modified with a defibrotizing compound as described herein (e.g., a compound of formula (I)). In some embodiments, at least one polymer in the blend comprising the second (outer) compartment is covalently modified with both a photoactive crosslinker and a defibrotizing compound.

[0340] In some embodiments, the first compartment comprises a blend of 2, 3, 4, 5, 6, 7, 8, 9, 10 or more polymers. In some embodiments, the first compartment comprises a blend of 2 polymers. In some embodiments, the first compartment comprises a blend of 3 polymers. In some embodiments, the first compartment comprises a blend of 4 polymers. In some embodiments, the first compartment comprises a blend of 5 polymers. In some embodiments, the first compartment comprises a blend of 6 polymers. In some embodiments, the first (inner) compartment comprises a blend of 7 polymers. In some embodiments, the first (inner) compartment comprises a blend of 8 polymers. In some embodiments, the first (inner) compartment comprises a blend of 9 polymers. In some embodiments, the first (inner) compartment comprises a blend of 10 polymers.

[0341] In some embodiments, the second (outer) compartment comprises a blend of 2, 3, 4, 5, 6, 7, 8, 9, 10 or more polymers. In some embodiments, the second (outer) compartment comprises a blend of 2 polymers. In some embodiments, the second (outer) compartment comprises a blend of 3 polymers. In some embodiments, the second (outer) compartment comprises a blend of 4 polymers. In some embodiments, the second (outer) compartment comprises a blend of 5 polymers. In some embodiments, the second (outer) compartment comprises a blend of 6 polymers. In some embodiments, the second (outer) compartment comprises a blend of 7 polymers. In some embodiments, the second (outer) compartment comprises a blend of 8 polymers. In some embodiments, the second (outer) compartment comprises a blend of 9 polymers. In some embodiments, the second (outer) compartment comprises a blend of 10 polymers.

[0342] In some embodiments, the first compartment comprises a blend of polymers and the second compartment does not comprise a blend of polymers. In some embodiments, the first compartment comprises a blend of polymers and the second compartment comprises a single type of polymer.

[0343] In some embodiments, the first compartment does not contain a blend of polymers and the second compartment contains a blend of polymers. In some embodiments, the first compartment contains a single type of polymer and the second compartment contains a blend of polymers.

[0344] In some embodiments, the first and second compartments comprise a blend of polymers, and the polymers of the blend are any two miscible polymers.

[0345] In some embodiments, the first and second compartments comprise a blend of polymers, and the polymer is selected from the group consisting of alginate, hyaluronate, and chitosan.

[0346] In some embodiments, the first and second compartments comprise a blend of polymers selected from the group consisting of alginate, hyaluronate, and chitosan. In some embodiments, the first compartment comprises a blend of polymers selected from the group consisting of alginate, hyaluronate, and chitosan. In some embodiments, the second compartment comprises a blend of polymers selected from the group consisting of alginate, hyaluronate, and chitosan.

[0347] In some embodiments, the first and second compartments comprise a blend of alginate polymers. In some embodiments, the first compartment comprises a blend of alginate polymers. In some embodiments, the second compartment comprises a blend of alginate polymers.

[0348] In some embodiments, the first and second compartments comprise a blend of alginate polymers, and the alginate polymer is selected from high guluronic acid alginate and high mannuronic acid alginate. In some embodiments, the first compartment comprises a blend of alginate polymers, and the alginate polymer is selected from high guluronic acid alginate and high mannuronic acid alginate. In some embodiments, the second compartment comprises a blend of alginate polymers, and the alginate polymer is selected from high guluronic acid alginate and high mannuronic acid alginate.

[0349] In some embodiments, the first and second compartments comprise a blend of alginate polymers, and this alginate polymer is selected from low molecular weight alginate, medium molecular weight alginate, high molecular weight alginate and ultra-high molecular weight alginate. In some embodiments, the first compartment comprises a blend of alginate polymers, and this alginate polymer is selected from low molecular weight alginate, medium molecular weight alginate, high molecular weight alginate and ultra-high molecular weight alginate. In some embodiments, the second compartment comprises a blend of alginate polymers, and this alginate polymer is selected from low molecular weight alginate, medium molecular weight alginate, high molecular weight alginate and ultra-high molecular weight alginate.

[0350] In some embodiments, the first and second compartments comprise a blend of alginate polymers, and the alginate polymer is selected from the group consisting of Kimica Algin IL-2, Kimica Algin IL-6, Kimica Algin 1-1, Kimica Algin 1-3, Kimica Algin 1-5, Kimica Algin 1-8, Kimica Algin LZ-2, Kimica Algin ULV-L3, Kimica Algin ULV-L5, Kimica Algin ULV-1G, Kimica Algin ULV-5G, Kimica Algin ULVIL-6G, Pronova UP VLVM, Pronova UP LVM, Pronova UP MVM, Pronova UP VLVG, PronovaUP MVG, Pronova UP LVG, Pronova SLM20, Pronova SLM100, Pronova SLG20, and PronovaSLG100. In some embodiments, the first compartment contains a blend of alginate polymers, and the alginate polymer is selected from Kimica Algin IL-2, Kimica Algin IL-6, Kimica Algin 1-1, Kimica Algin 1-3, Kimica Algin 1-5, Kimica Algin 1-8, Kimica Algin LZ-2, Kimica Algin ULV-L3, Kimica Algin ULV-L5, Kimica Algin ULV-1G, Kimica Algin ULV-5G, Kimica Algin ULVIL-6G, Pronova UP VLVM, Pronova UP LVM, Pronova UP MVM, Pronova UP VLVG, PronovaUP MVG, Pronova UP LVG, Pronova SLM20, Pronova SLM100, Pronova SLG20, and PronovaSLG100.In some embodiments, the second compartment comprises a blend of alginate polymers, and the alginate polymer is selected from Kimica Algin IL-2, Kimica Algin IL-6, Kimica Algin 1-1, Kimica Algin 1-3, Kimica Algin 1-5, Kimica Algin 1-8, Kimica Algin LZ-2, Kimica Algin ULV-L3, Kimica Algin ULV-L5, Kimica Algin ULV-1G, Kimica Algin ULV-5G, Kimica Algin ULVIL-6G, Pronova UP VLVM, Pronova UP LVM, Pronova UP MVM, Pronova UP VLVG, PronovaUP MVG, Pronova UP LVG, Pronova SLM20, Pronova SLM100, Pronova SLG20, and PronovaSLG100.

[0351] In some embodiments, the first and second compartments include a blend of two alginate polymers in any ratio. In some embodiments, the ratio of the two alginate polymers in the blend is about 99:1, 95:5, 90:10, 85:15, 80:20, 75:25, 70:30, 65:35, 60:40, 55:45 or 50:50. In some embodiments, the ratio of the two alginate polymers in the blend is about 99:1. In some embodiments, the ratio of the two alginate polymers in the blend is about 95:5. In some embodiments, the ratio of the two alginate polymers in the blend is about 90:10. In some embodiments, the ratio of the two alginate polymers in the blend is about 85:15. In some embodiments, the ratio of the two alginate polymers in the blend is about 80:20. In some embodiments, the ratio of the two alginate polymers in the blend is about 75:25. In some embodiments, the ratio of the two alginate polymers in the blend is about 70:30. In some embodiments, the ratio of the two alginate polymers in the blend is about 65:35. In some embodiments, the ratio of the two alginate polymers in the blend is about 60:40. In some embodiments, the ratio of the two alginate polymers in the blend is about 55:45. In some embodiments, the ratio of the two alginate polymers in the blend is about 50:50.

[0352] In some embodiments, the first and second compartments comprise a blend of two alginate polymers in any ratio. In some embodiments, the ratio of the two alginate polymers in the blend is greater than about 99:1, 95:5, 90:10, 85:15, 80:20, 75:25, 70:30, 65:35, 60:40, 55:45 or 50:50. In some embodiments, the ratio of the two alginate polymers in the blend is greater than about 99:1. In some embodiments, the ratio of the two alginate polymers in the blend is greater than about 95:5. In some embodiments, the ratio of the two alginate polymers in the blend is greater than about 90:10. In some embodiments, the ratio of the two alginate polymers in the blend is greater than about 85:15. In some embodiments, the ratio of the two alginate polymers in the blend is greater than about 80:20. In some embodiments, the ratio of the two alginate polymers in the blend is greater than about 75:25. In some embodiments, the ratio of the two alginate polymers in the blend is greater than about 70:30. In some embodiments, the ratio of the two alginate polymers in the blend is greater than about 65:35. In some embodiments, the ratio of the two alginate polymers in the blend is greater than about 60:40. In some embodiments, the ratio of the two alginate polymers in the blend is greater than about 55:45. In some embodiments, the ratio of the two alginate polymers in the blend is greater than about 50:50.

[0353] In some embodiments of the invention, the first and second compartments comprise a blend of VLVG alginate and SLG100 alginate. In some embodiments of the invention, the first compartment comprises a blend of VLVG alginate and SLG100 alginate. In some embodiments of the invention, the second compartment comprises a blend of VLVG alginate and SLG100 alginate.

[0354] In some embodiments of the invention, the first and second compartments comprise a blend of VLVG alginate and SLG100 alginate. In some embodiments of the invention, the first compartment comprises a blend of VLVG alginate and SLG100 alginate. In some embodiments of the invention, the second compartment comprises a blend of VLVG alginate and SLG100 alginate.

[0355] In some embodiments, the polymer in one or both of the first and second compartments is: (i) a low molecular weight alginate, e.g., having an approximate MW of <75 kDa and a G:M ratio of ≥1.5; (ii) a medium molecular weight alginate, e.g., having an approximate molecular weight of 75 to 150 kDa and a G:M ratio of ≥1.5; (iii) a high molecular weight alginate, e.g., having an approximate MW of 150 kDa to 250 kDa and a G:M ratio of ≥1.5; (iv) or a blend of two or more of these alginates. In one embodiment, the polymer in the first (inner) compartment is an unmodified high molecular weight alginate or an unmodified medium molecular weight alginate, and the polymer in the second (outer) compartment is a blend of chemically modified alginate (e.g., alginate modified with Compound 101 shown in Table 3) and unmodified alginate, such as a 70:30 blend or a 60:40 blend of CM-LMW-Alg-101:U-HMW-Alg, which can be prepared as described in the Examples below.

[0356] In some embodiments, the particle (e.g., hydrogel capsule) comprises alginate, and the compound of formula (I) is covalently attached to some or all of the monomers in the alginate. In some embodiments, some or all of the monomers in the alginate are modified by the same compound of formula (I). In some embodiments, some or all of the monomers in the alginate are modified by different compounds of formula (I).

[0357] In some embodiments, the polymer of the first compartment of the particle (e.g., a hydrogel capsule) is modified with one compound of formula (I), and the polymer of the second compartment of the particle (e.g., a hydrogel capsule) is modified with a different compound of formula (I). In some embodiments, the particle (e.g., a hydrogel capsule) comprises a mixture of a polymer modified with a compound of formula (I) and an unmodified polymer (e.g., a polymer not modified with a compound of formula (I)). In some embodiments, the first compartment comprises a mixture (i.e., a blend) of a polymer modified with a compound of formula (I) and an unmodified polymer (e.g., a polymer not modified with a compound of formula (I)). In some embodiments, the second compartment comprises a mixture of a polymer modified with a compound of formula (I) and an unmodified polymer (e.g., a polymer not modified with a compound of formula (I)).

[0358] The polymer of the particles described herein (e.g., hydrogel capsules) can be modified with a compound of formula (I) or a pharmaceutically acceptable salt thereof on one or more monomers of the polymer. The modified polymer of the particle (e.g., hydrogel capsule) can be present in the first (inner) compartment of the particle, the second (outer) compartment of the particle, or both the first (inner) compartment and the second (outer) compartment of the particle. In some embodiments, the modified polymer is only present in the second compartment (which includes the outer particle surface). In some embodiments, at least 0.5% of the monomers of the polymer are modified with a compound of formula (I) (e.g., at least 1%, 2.5%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or more of the monomers of the polymer are modified with a compound of formula (I)). In some embodiments, 0.5% to 50%, 10% to 90%, 10% to 50%, or 25% to 75% of the monomers of the polymer are modified with a compound of formula (I). In some embodiments, 1% to 20% of the monomers of the polymer are modified with compounds of formula (I). In some embodiments, 1% to 10% of the monomers of the polymer are modified with compounds of formula (I).

[0359] In some embodiments, the polymer (e.g., alginate) when modified with a compound of Formula (I), e.g., compound 101 of Table 3, comprises an increase in % N of any of the following values ​​(compared to the unmodified polymer (e.g., alginate): (i) at least 0.1%, 0.2%, 0.5%, 1.0%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% N by weight; (ii) 0.1% to 10% N by weight; (iii) 0.1% to 2% N by weight; (iv) 2% to 4% N by weight; (v) 4% to 8% N by weight; (vi) 5% to 9% N by weight; (vii) 6% to 9% N by weight; (viii) 6% to 8% N by weight; (ix) 7% to 9% N by weight. N; and (x) 8 to 9% by weight N, wherein in each case the % N is determined by combustion analysis (e.g. as described herein in Example 2) and corresponds to the amount of the compound of formula (I) in the modified polymer.

[0360] Particle (for example, hydrogel capsule) (for example, the first compartment or the second compartment therein) can include the formula (I) compound of the amount giving the particle specific characteristics.For example, particle surface (for example, the outside of the outer compartment) can include so that particle has the concentration or density of the formula (I) compound of defibrosis (that is, alleviating foreign body reaction) in subject.In one embodiment, particle surface includes the alginate chemically modified by compound 101 of defibrosis effective amount.In one embodiment, the compound 101 of defibrosis effective amount produces about 0.5% to 2%, 2% to 4% N, about 4% to 6% N, about 6% to 8% or about 8% to 10% N% N increase (compared to unmodified alginate), wherein% N is determined by combustion analysis (for example, as described in Example 2 herein) and corresponds to the amount of the compound 101 in modified alginate.

[0361] In one embodiment, a 5 mm probe attached to a 5 kg load cell is used to perform mechanical testing of hydrogel capsules on a TA.XT plus texture analyzer (Stable Micro Systems, Surrey, United Kingdom). A single capsule is placed on a platform and compressed from above by a probe at a fixed rate of 0.5 mm / s. When a repulsive force of 1 g is measured, contact between the probe and the capsule is detected. The probe continues to advance 90% of the distance between the probe's contact height and the platform, thereby compressing the capsule to its bursting point. The probe's resistance to compression is measured and plotted as a function of probe travel (force versus displacement curve). Typically, before the capsule bursts completely, the capsule will slightly rupture, and the force applied to the probe will be reduced by a small amount. The analysis macro can be programmed to detect a decrease in the force versus displacement curve that occurs for the first time, which is 0.25-0.5 g. The force applied by the probe when this occurs is referred to as the initial rupture force. In one embodiment, the desired mechanical strength of the particles described herein (eg, two-compartment hydrogel capsules) has an initial rupture force greater than 1, 1.5, 2, 2.5, or 3 grams, or at least 2 grams.

[0362] In one embodiment, the desired mechanical strength of a particle (e.g., a hydrogel capsule) is the ability to remain intact at a desired time point after implantation in a subject, e.g., the outer and inner compartments of a hydrogel capsule removed from a subject are visibly intact after retrieval from an immunocompetent mouse when observed by optical microscopy, e.g., by bright field imaging as described in the Examples herein.

[0363] In one embodiment, the surface of the particle comprises alginate chemically modified with Compound 101 in an amount that provides defibrotizing properties and desired mechanical strength to the particle, for example, the concentration or density of Compound 101 in the modified alginate produces an increase in % N (compared to unmodified alginate) of any of the following values: (i) 1% to 3% by weight; (ii) 2% to 4% N by weight; (iii) 4% to 8% N by weight; (iv) 5% to 9% N by weight; (v) 6% to 9% N by weight; (vi) 6% to 8% N by weight; (vii) 7% to 9% N by weight; and (ix) 8% to 9% N by weight; wherein in each case, % N is determined by combustion analysis (e.g., as described in Example 2 herein) and corresponds to the amount of compound of formula (I) in the modified alginate.

[0364] When the particles (e.g., the first compartment or the second compartment thereof) contain alginate, the alginate can be chemically modified with a compound of formula (I) using any suitable method known in the art. For example, the alginate carboxylic acid moiety can be activated to couple with one or more amine-functionalized compounds to obtain alginate modified with a compound of formula (I). The alginate polymer can be dissolved in water (30 ml / g polymer) and treated with 2-chloro-4,6-dimethoxy-1,3,5-triazine (0.5 eq) and N-methylmorpholine (1 eq). A solution (0.3 M) of the compound of formula (I) dissolved in a buffer or solvent (such as acetonitrile) can be added to this mixture. The reactants can be heated to, for example, 55°C and kept for 16 hours, then cooled to room temperature and concentrated via rotary evaporation. The residue can then be dissolved in a buffer or solvent (e.g., water). The mixture can then be filtered, for example, through a cyano-modified silica gel bed (Silicycle), and the filter cake can be washed with water. The resulting solution can then be dialyzed (10,000 MWCO membrane) against buffer or water for 24 hours, eg, exchanging the buffer or water at least once, at least twice, at least three times, or more.The resulting solution can be concentrated, eg, via lyophilization, to yield the desired chemically modified alginate.

[0365] In some embodiments, the particles described herein include cells. In some embodiments, the cells are engineered to produce therapeutic agents (e.g., proteins or polypeptides, such as antibodies, proteins, enzymes, or growth factors). In some embodiments, the cells are placed in the first compartment. In some embodiments, the cells are placed in the second compartment. In some embodiments, the cells are placed in the first compartment and the second compartment does not include cells. Particles (e.g., hydrogel capsules) may include active or inactive fragments of proteins or polypeptides, such as glucose oxidase (e.g., for glucose sensors), kinases, phosphatases, oxygenases, hydrogenases, reductases.

[0366] The particles described herein (e.g., hydrogel capsules) can be configured to release therapeutic agents, such as exogenous substances, such as therapeutic agents described herein. In some embodiments, the therapeutic agent is a compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the therapeutic agent is a biomaterial. In some embodiments, the therapeutic agent is a nucleic acid (e.g., RNA or DNA), a protein (e.g., a hormone, an enzyme, an antibody, an antibody fragment, an antigen or an epitope), a small molecule, a lipid, a drug, a vaccine, or any derivative thereof.

[0367] Particles (e.g., hydrogel capsules) (e.g., as described herein) can be provided in the form of a formulation or composition for implantation or administration to a subject. In some embodiments, at least 20%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% of the particles (e.g., hydrogel capsules) in the formulation or composition have characteristics as described herein, such as average diameter or average pore size.

[0368] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising a blend of VLVG alginate and SLG100 alginate, wherein the VLVG alginate comprises Compound 101 and the SLG100 alginate is unmodified; and (ii) an outer compartment comprising a blend of VLVG alginate and SLG100 alginate, wherein the VLVG alginate comprises Compound 101 and the SLG100 alginate is unmodified.

[0369] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising a blend of VLVG alginate and SLG100 alginate, wherein the VLVG alginate comprises Compound 101 and the SLG100 alginate comprises Compound 205; and (ii) an outer compartment comprising a blend of VLVG alginate and SLG100 alginate, wherein the VLVG alginate comprises Compound 101 and the SLG100 alginate comprises Compound 205.

[0370] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 200.

[0371] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 201.

[0372] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 202.

[0373] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 203.

[0374] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 204.

[0375] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 205.

[0376] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 214.

[0377] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 215.

[0378] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 216.

[0379] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 217.

[0380] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 218.

[0381] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 219.

[0382] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 200.

[0383] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 201.

[0384] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 202.

[0385] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 203.

[0386] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 204.

[0387] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 205.

[0388] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 214.

[0389] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 215.

[0390] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 216.

[0391] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 217.

[0392] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 218.

[0393] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 219.

[0394] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 101 and Compound 201.

[0395] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 101 and Compound 202.

[0396] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 101 and Compound 203.

[0397] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 101 and Compound 204.

[0398] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 101 and Compound 205.

[0399] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 101 and Compound 214.

[0400] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 101 and Compound 215.

[0401] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 101 and Compound 216.

[0402] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 101 and Compound 217.

[0403] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 101 and Compound 218.

[0404] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 101 and Compound 219.

[0405] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 200.

[0406] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 201.

[0407] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 202.

[0408] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 203.

[0409] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 204.

[0410] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 205.

[0411] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 214.

[0412] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 215.

[0413] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 216.

[0414] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 217.

[0415] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 218.

[0416] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 219.

[0417] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 200.

[0418] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 201.

[0419] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 202.

[0420] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 203.

[0421] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 204.

[0422] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 205.

[0423] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 214.

[0424] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 215.

[0425] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 216.

[0426] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 217.

[0427] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 218.

[0428] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate; and (ii) an outer compartment comprising LG20 comprising Compound 114 and Compound 219.

[0429] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 114 and Compound 201.

[0430] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 114 and Compound 202.

[0431] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 114 and Compound 203.

[0432] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 114 and Compound 204.

[0433] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 114 and Compound 205.

[0434] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 114 and Compound 214.

[0435] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 114 and Compound 215.

[0436] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 114 and Compound 216.

[0437] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 114 and Compound 217.

[0438] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 114 and Compound 218.

[0439] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate; and (ii) an outer compartment comprising SLG100 comprising Compound 114 and Compound 219.

[0440] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG20 alginate; and (ii) an outer compartment comprising LG20 alginate modified with Compound 101 and Compound 205.

[0441] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG20 alginate; and (ii) an outer compartment comprising LG20 alginate modified with Compound 101 and Compound 200.

[0442] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG20 alginate; and (ii) an outer compartment comprising LG20 alginate modified with Compound 101 and Compound 201.

[0443] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG20 alginate; and (ii) an outer compartment comprising LG20 alginate modified with Compound 101 and Compound 202.

[0444] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG20 alginate; and (ii) an outer compartment comprising LG20 alginate modified with Compound 101 and Compound 203.

[0445] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG20 alginate; and (ii) an outer compartment comprising LG20 alginate modified with Compound 101 and Compound 204.

[0446] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG20 alginate; and (ii) an outer compartment comprising LG20 alginate modified with Compound 101 and Compound 206.

[0447] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG20 alginate; and (ii) an outer compartment comprising LG20 alginate modified with Compound 114 and Compound 205.

[0448] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG20 alginate; and (ii) an outer compartment comprising LG20 alginate modified with Compound 114 and Compound 200.

[0449] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG20 alginate; and (ii) an outer compartment comprising LG20 alginate modified with Compound 114 and Compound 201.

[0450] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG20 alginate; and (ii) an outer compartment comprising LG20 alginate modified with Compound 114 and Compound 202.

[0451] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG20 alginate; and (ii) an outer compartment comprising LG20 alginate modified with Compound 114 and Compound 203.

[0452] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG20 alginate; and (ii) an outer compartment comprising LG20 alginate modified with Compound 114 and Compound 204.

[0453] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG20 alginate; and (ii) an outer compartment comprising LG20 alginate modified with Compound 114 and Compound 206.

[0454] Cells and therapeutic agents

[0455] The hydrogel or hydrogel capsule of the present invention may include a variety of different cell types (e.g., human cells), including but not limited to: adipocytes, epidermal cells, epithelial cells, endothelial cells, fibroblasts, pancreatic islet cells, mesenchymal stem cells, pericytes, a subtype of any of the foregoing, a cell derived from any of the foregoing, a cell derived from induced pluripotent stem cells, and a mixture of one or more of any of the foregoing. Exemplary cell types include the cell types described in WO 2017 / 075631 and WO 2019 / 195055. In one embodiment, the hydrogel capsule described herein includes a plurality of cells. In one embodiment, a plurality of cells are in the form of a cell suspension before being encapsulated in a hydrogel capsule described herein. The cells in the suspension can take the form of a single cell (e.g., from a monolayer cell culture), or in another form (e.g., placed on a microcarrier (e.g., beads or matrix)) or as a three-dimensional aggregate of cells (e.g., cell clusters or spheroids) provided. The cell suspension may comprise a plurality of cell clusters (eg, as spheroids) or microcarriers.In some embodiments, the hydrogel capsule does not comprise any pancreatic islet cells and does not comprise any cells capable of producing insulin in a glucose-responsive manner.

[0456] In some embodiments, compared to an untreated control group (e.g., a substantially identical hydrogel capsule lacking a compound of formula (I), the hydrogel or hydrogel capsule of the present disclosure reduces immune cell adhesion. In one embodiment, compared to an untreated control, the hydrogel capsule reduces macrophage adhesion. In one embodiment, compared to an untreated control, macrophage adhesion is reduced by about 1 to 10 times. In one embodiment, compared to an untreated control, macrophage adhesion is reduced by about 1 to 8 times. In one embodiment, compared to an untreated control, macrophage adhesion is reduced by about 1 to 7 times. In one embodiment, compared to an untreated control, macrophage adhesion is reduced by about 1 to 6 times. In one embodiment, compared to an untreated control, macrophage adhesion is reduced by about 1 to 5 times.

[0457] The hydrogels or hydrogel capsules of the present disclosure allow encapsulated cells (e.g., engineered cells) to retain viability (e.g., as determined by a cell viability assay). In some embodiments, the hydrogels or hydrogel capsules allow encapsulated cells to retain viability for at least seven days, at least one month, or at least one year.

[0458] The present disclosure is characterized in that a kind of through encapsulated cell, it produces or can produce for preventing or treating disease described herein, disease or illness therapeutic agent.In one embodiment, cell is naturally occurring cell, for example, without engineering.In one embodiment, cell is through engineered cell.In one embodiment, cell is engineered to sense stimulus, for example chemical signal, and expresses therapeutic agent in response to this stimulus.In one embodiment, cell is differentiated from stem cell (for example inducible rich potential stem cell), and differentiated cell can produce therapeutic agent continuously or in response to stimulus.The therapeutic agent produced by through encapsulated cell can be any biological substance, such as nucleic acid (for example nucleotide, DNA or RNA), polypeptide, lipid, sugar (for example monosaccharide, disaccharide, oligosaccharide or polysaccharide) or small molecule, each of which is described in further detail below.Exemplary therapeutic agent includes the agent listed in WO 2017 / 075631 and WO 2019 / 195055.

[0459] In some embodiments, nucleic acid is produced through encapsulated cells (e.g., engineered cells). The nucleic acid produced by cells described herein can vary in size and contain one or more nucleosides or nucleotides, for example, more than 2, 3, 4, 5, 10, 25, 50 or more nucleosides or nucleotides. In some embodiments, nucleic acid is a short fragment of, for example, RNA or DNA, and can be used as a reporter gene or for diagnostic purposes. Exemplary nucleic acid includes single nucleosides or nucleotides (e.g., adenosine, thymidine, cytidine, guanosine, uridine monophosphate, inosine monophosphate), RNA (e.g., mRNA, siRNA, miRNA, RNAi) and DNA (e.g., vector, chromosomal DNA). In some embodiments, nucleic acid has an average molecular weight (kD) of about 0.25, 0.5, 1, 1.5, 2, 2.5, 5, 10, 25, 50, 100, 150, 200 or more.

[0460] In some embodiments, the therapeutic agent is a peptide or polypeptide (e.g., protein), such as a hormone, enzyme, cytokine (e.g., proinflammatory cytokine or anti-inflammatory cytokine), growth factor, coagulation factor or lipoprotein. The peptide or polypeptide (e.g., protein, such as a hormone, growth factor, coagulation factor or coagulation factor, antibody molecule, enzyme, cytokine, cytokine receptor or chimeric protein including a cytokine or cytokine receptor) produced by the cells in the implantable element (e.g., hydrogel capsule described herein) may have a naturally occurring amino acid sequence, or may contain a variant of a naturally occurring sequence. The variant may be a naturally occurring or non-naturally occurring amino acid substitution, mutation, deletion or addition relative to a reference naturally occurring sequence. The naturally occurring amino acid sequence may be a polymorphic variant. The naturally occurring amino acid sequence may be a human or non-human amino acid sequence. In some embodiments, the naturally occurring amino acid sequence or its naturally occurring variant is a human sequence. In addition, the peptide or polypeptide (e.g., protein) used in the present invention may be modified in some way, such as by chemical or enzymatic modification (e.g., glycosylation, phosphorylation). In some embodiments, the peptide has about 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 45, or 50 amino acids. In some embodiments, the protein has an average molecular weight (kD) of 5, 10, 25, 50, 100, 150, 200, 250, 500, or more.

[0461] In some embodiments, protein is a hormone. Exemplary hormones include antidiuretic hormone (ADH), oxytocin, growth hormone (GH), prolactin, growth hormone-releasing hormone (GHRH), thyroid stimulating hormone (TSH), thyrotropin-releasing hormone (TRH), adrenocorticotropic hormone (ACTH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), luteinizing hormone-releasing hormone (LHRH), thyroxine, calcitonin, parathyroid hormone, aldosterone, cortisol, epinephrine, glucagon, insulin, estrogen, progesterone and testosterone. In some embodiments, protein is insulin (e.g., insulin A chain, insulin B chain or proinsulin). In some embodiments, protein is growth hormone, such as human growth hormone (hGH), recombinant human growth hormone (rhGH), bovine growth hormone, methionine-human growth hormone, dephenylalanine human growth hormone and porcine growth hormone. In some embodiments, protein is not insulin (e.g., insulin A chain, insulin B chain or proinsulin).

[0462] In some embodiments, the protein is a growth factor, such as vascular endothelial growth factor (VEGF), nerve growth factor (NGF), platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), epidermal growth factor (EGF), transforming growth factor (TGF), and insulin-like growth factor-I and -II (IGF-I and IGF-II).

[0463] In some embodiments, the protein is a clotting factor or coagulation factor, such as a blood clotting factor or blood coagulation factor. In some embodiments, the protein is a protein that participates in coagulation, the process by which blood changes from a liquid to a solid or gel. Exemplary coagulation factors and clotting factors include factor I (e.g., fibrinogen), factor II (e.g., prothrombin), factor III (e.g., tissue factor), factor V (e.g., prothrombin, unstable factor), factor VI, factor VII (e.g., stable factor, proconvertin), factor VIII (e.g., antihemophilic factor A), factor VIIIC, factor IX (e.g., antihemophilic factor B), factor X (e.g., Stuart-Prower factor), factor XI (e.g., plasma thromboplastin precursor), factor XII (e.g., Hagemann factor), factor XIII (e.g., fibrin stabilizing factor), von Willebrand factor, prekallikrein, heparin cofactor II, high molecular weight kininogen (e.g., Fitzgerald factor), antithrombin III, and fibronectin. In some embodiments, the protein is an anti-coagulation factor, such as protein C.

[0464] In some embodiments, protein is an antibody molecule. As used herein, the term "antibody molecule" refers to a protein, such as an immunoglobulin chain or a fragment thereof comprising at least one immunoglobulin variable domain sequence. The term "antibody molecule" includes, for example, a monoclonal antibody (including a full-length antibody with an immunoglobulin Fc region). In one embodiment, the antibody molecule comprises a full-length antibody or a full-length immunoglobulin chain. In one embodiment, the antibody molecule comprises an antigen-binding or functional fragment of a full-length antibody or a full-length immunoglobulin chain. In one embodiment, the antibody molecule is a monospecific antibody molecule and binds to a single epitope, such as a monospecific antibody molecule with a plurality of immunoglobulin variable domain sequences, each immunoglobulin variable domain sequence binds to the same epitope.

[0465] In one embodiment, antibody molecule is a multispecific antibody molecule, for example, it comprises a plurality of immunoglobulin variable domain sequences, wherein the first immunoglobulin variable domain sequence in the plurality of immunoglobulin variable domain sequences has binding specificity to the first epitope, and the second immunoglobulin variable domain sequence in the plurality of immunoglobulin variable domain sequences has binding specificity to the second epitope. In one embodiment, the first epitope and the second epitope are in the same antigen, for example, on the same protein (or subunit of a multimeric protein). In one embodiment, the multispecific antibody molecule comprises the third, fourth or fifth immunoglobulin variable domain. In one embodiment, the multispecific antibody molecule is a bispecific antibody molecule, a trispecific antibody molecule or a tetraspecific antibody molecule.

[0466] Various types of antibody molecules can be produced by cells in implantable elements (e.g., hydrogel capsules) as described herein, including complete immunoglobulins of any class, fragments thereof, and synthetic proteins containing at least the antigen-binding variable domains of antibodies. Antibody molecules can be antibodies, such as IgG antibodies, such as IgG1, IgG2, IgG3, or IgG4. Antibody molecules can be in the form of antigen-binding fragments, including Fab fragments, F(ab')2 fragments, single-chain variable regions, etc. Antibodies can be polyclonal or monoclonal (mAbs). Monoclonal antibodies can include "chimeric" antibodies, in which a portion of a heavy chain and / or a light chain is identical or homologous to the corresponding sequence in an antibody derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain is identical or homologous to the corresponding sequence in an antibody derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, as long as they specifically bind to the target antigen and / or exhibit desired biological activity. In some embodiments, the antibody molecule is a single domain antibody (e.g., nanobody). Antibodies can also be modified by recombinant means, for example, by deletion, addition or substitution of amino acids, to increase the efficacy of the antibody in mediating the desired function. Exemplary antibodies include anti-β-galactosidase, anti-collagen, anti-CD14, anti-CD20, anti-CD40, anti-HER2, anti-IL-1, anti-IL-4, anti-IL6, anti-IL-13, anti-IL17, anti-IL18, anti-IL-23, anti-IL-28, anti-IL-29, anti-IL-33, anti-EGFR, anti-VEGF, anti-CDF, anti-flagellin, anti-IFN-α, anti-IFN-β, anti-IFN-γ, anti-mannose receptor, anti-VEGF, anti-TLR1, anti-TLR2, anti-TLR3, anti-TLR4, anti-TLR5, anti-TLR6, anti-TLR9, anti-PDF, anti-PD1, anti-PDL-1 or anti-nerve growth factor antibodies. In some embodiments, the antibody is an anti-nerve growth factor antibody (eg, voranuzumab, fasitumab, tanezumab).

[0467] In some embodiments, the protein is a cytokine or cytokine receptor, or a chimeric protein comprising a cytokine or its receptor, including, for example, tumor necrosis factor alpha and beta, their receptors and their derivatives, renin; lipoproteins; colchicine; adrenocorticotropic hormone; vasopressin; somatostatin; lysine vasopressin; cholecystokinin; leuprolide; alpha-1-antitrypsin; atrial natriuretic factor; pulmonary surfactant; plasminogen activators other than tissue-type plasminogen activator (t-PA), such as urokinase; bombesin; thrombin; neprilysin; RANTES (regulates activation, normal T-cell expression and secretion); human macrophage inflammatory protein (MIP-1-α); serum albumin, such as human serum albumin; Müllerian inhibitory substance; relaxin A chain; relaxin B chain; prorelaxin; mouse gonadotropin-related peptide; chorionic gonadotropin; microbial proteins, such as beta-lactamase; DNA enzymes; inhibins; activins; receptors for hormones or growth factors; integrins; proteins A or D; rheumatoid factor; platelet-derived growth factor (PDGF); epidermal growth factor (EGF); transforming growth factors (TGFs), such as TGF-α and TGF-β, including TGF-β1, TGF-β2, TGF-β3, TGF-β4, or TGF-β5; insulin-like growth factors-I and -II (IGF-I and IGF-II); des(1-3)-IGF-I (brain IGF-I), insulin-like growth factor binding protein; CD proteins such as CD-3, CD-4, CD-8 and CD-19; erythropoietin; osteoinductive factors; immunotoxins; interferons such as interferon-α (e.g., interferon α.2A), interferon-β, interferon-γ, interferon-λ and consensus interferon; colony stimulating factors (CSFs), such as M-CSF, GM-CSF and G-CSF; interleukins (ILs), such as IL-1 to IL-10; superoxide dismutase; T cell receptors; surface membrane proteins; decay accelerating factors; transporters; homing receptors; addressins; fertility inhibitors such as prostaglandins; fertility promoters; regulatory proteins; antibodies (including fragments thereof) and chimeric proteins such as immunoadhesins; precursors, derivatives, prodrugs and analogs of these compounds, as well as pharmaceutically acceptable salts of these compounds or their precursors, derivatives, prodrugs and analogs. Suitable proteins or peptides can be natural or recombinant and include, for example, fusion proteins.

[0468] Examples of polypeptides (e.g., proteins) produced by cells in an implantable element (e.g., a hydrogel capsule) described herein also include CCL1, CCL2 (MCP-1), CCL3 (MIP-1α), CCL4 (MIP-1β), CCL5 (RANTES), CCL6, CCL7, CCL8, CCL9 (CCL10), CCL11, CCL12, CCL13, CCL14, CCL15, CCL16, CCL17, CCL18, CCL19, CCL20, CCL21, CCL22, CCL23, CCL24, CCL25, CCL26, CCL27, CCL28, CXCL1 (KC), CXCL2 (SDF1a), CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, CXCL8 (IL8), CXCL9, CXCL10, CXCL11, CXCL12, CXCL13, CXCL14, CXCL15, CXCL16, CXCL17, CX3CL1, XCL1, XCL2, TNFA, TNFB (LTA), TNFC (LTB), TNFSF4, TNFSF5 (CD40LG), TNFSF6, TNFSF7, TNFSF8, TNFSF9, TNFSF10, TNFSF11, TNFSF13B, EDA, IL2, IL15, IL4, IL13, IL7, IL 9. IL21, IL3, IL5, IL6, IL11, IL27, IL30, IL31, OSM, LIF, CNTF, CTF1, IL12a, IL12b, IL23, IL27, IL35, IL14, IL 16. IL32, IL34, IL10, IL22, IL19, IL20, IL24, IL26, IL29, IFNL1, IFNL2, IFNL3, IL28, IFNA1, IFNA2, IFNA4, I FNA5, IFNA6, IFNA7, IFNA8, IFNA10, IFNA13, IFNA14, IFNA16, IFNA17, IFNA21, IFNB1, IFNK, IFNW1, IFNG, IL1A (IL1F1), IL1B (IL1F2), IL1Ra (IL1F3), IL1F5 (IL36RN), IL1F6 (IL36A), IL1F7 (IL37), IL1F8 (IL36B), IL1F9 (IL36G), IL1F10 (IL38), IL33 (IL1F11), IL18(IL1G), IL17, KITLG, IL25 (IL17E), CSF1 (M-CSF), CSF2 (GM-CSF), CSF3 (G-CSF), SPP1,TGFB1, TGFB2, TGFB3, CCL3L1, CCL3L2, CCL3L3, CCL4L1, CCL4L2, IL17B, IL17C, IL17D, IL17F, AIMP1 (SCYE1), MIF, Areg, BC096441, Bmp1, Bmp10, Bmp15, Bmp2, Bmp3, Bmp4, Bmp5, Bmp6, Bmp7, Bmp8a, Bmp8b, C1qtnf4, Ccl21a, Ccl27a, Cd70, Ce r1, Cklf, Clcf1, Cmtm2a, Cmtm2b, Cmtm3, Cmtm4, Cmtm5, Cmtm6, Cmtm7, Cmtm8, Crlf1, Ctf2, Ebi3, Edn1, Fam3b, Fasl, Fgf2, Flt3l, Gdf10, G df11, Gdf15, Gdf2, Gdf3, Gdf5, Gdf6, Gdf7, Gdf9, Gm12597, Gm13271, Gm13275, Gm13276, Gm13280, Gm13283, Gm2564, Gpi1, Grem1, Grem2, G rn, Hmgb1, Ifna11, Ifna12, Ifna9, Ifnab, Ifne, Il17a, Il23a, Il25, Il31, Iltifb, Inhba, Lefty1, Lefty2, Mstn, Nampt, Ndp, Nodal, Pf4, P glyrp1, Prl7d1, Scg2, Scgb3a1, Slurp1, Spp1, Thpo, Tnfsf10, Tnfsf11, Tnfsf12, Tnfsf13, Tnfsf13b, Tnfsf14, Tnfsf15, Tnfsf18, Tnfsf4, Tnfsf8, Tnfsf9, Tslp, Vegfa, Wnt1, Wnt2, Wnt5a, Wnt7a, Xcl1, epinephrine, melatonin, triiodothyronine, prostaglandins, leukotrienes, prostacyclin, thromboxane, islet amylin, Müllerian inhibitory factor or hormone, adiponectin, nephrotropin Adrenal cortex hormone, angiotensin, vasopressin, arginine vasopressin, atrial natriuretic peptide, brain natriuretic peptide, calcitonin, cholecystokinin, cortistatin, enkephalin, endothelin, erythropoietin, follicle-stimulating hormone, galanin, gastric inhibitory polypeptide, gastrin, ghrelin, glucagon, glucagon-like peptide-1, gonadotropin-releasing hormone, hepcidin, human chorionic gonadotropin, human placental lactogen, inhibin, somatomedin, leptin, lipotropin, melanocyte-stimulating hormone, motilin, orexin, oxytocin, pancreatic polypeptide, pituitary adenylate cyclase-activating peptide, relaxin, renin, secretin, somatostatin, thrombopoietin, thyrotropin, thyrotropin-releasing hormone,Vasoactive intestinal peptide, androgens, alpha-glucosidase (also known as acid maltase), glycogen phosphorylase, glycogen debranching enzyme, phosphofructokinase, phosphoglycerate kinase, phosphoglycerate mutase, lactate dehydrogenase, carnitine palmitoyltransferase, carnitine and myoadenylate deaminase.

[0469] In some embodiments, the protein is an alternative therapy or alternative protein. In some embodiments, the alternative therapy or alternative protein is a coagulation factor or a clotting factor, such as Factor VIII (e.g., comprising a naturally occurring human Factor VIII amino acid sequence or a variant thereof) or Factor IX (e.g., comprising a naturally occurring human Factor IX amino acid sequence or a variant thereof).

[0470] In some embodiments, the encapsulated cells are engineered to express Factor VIII, e.g., recombinant Factor VIII. In some embodiments, the cells are derived from human tissue and are engineered to express Factor VIII, e.g., recombinant Factor VIII. In some embodiments, the recombinant Factor VIII is a recombinant Factor VIII with a B domain deleted (FVIII-BDD).

[0471] In some embodiments, the encapsulated cells are derived from human tissue and are engineered to express factor IX, e.g., recombinant factor IX. In some embodiments, the cells are engineered to express factor IX, e.g., wild-type human factor IX (FIX) or a polymorphic variant thereof. In some embodiments, the cells are engineered to express a gain-of-function (GIF) variant of a wild-type FIX protein (FIX-GIF), wherein the GIF variant has a higher specific activity than the corresponding wild-type FIX.

[0472] In some embodiments, the replacement therapy or replacement protein is an enzyme, such as α-galactosidase, α-L-iduronidase (IDUA) or N-sulfoglucosamine sulfohydrolase (SGSH). In some embodiments, the replacement therapy or replacement protein is an enzyme, such as α-galactosidase A (e.g., comprising a naturally occurring human α-galactosidase A amino acid sequence or a variant thereof). In some embodiments, the replacement therapy or replacement protein is a cytokine or an antibody.

[0473] In some embodiments, the therapeutic agent is a sugar, such as a monosaccharide, disaccharide, oligosaccharide or polysaccharide. In some embodiments, sugar includes triose, tetrose, pentose, hexose or heptose moieties. In some embodiments, sugar includes linear monosaccharides or cyclized monosaccharides. In some embodiments, sugar includes glucose, galactose, fructose, rhamnose, mannose, arabinose, glucosamine, galactosamine, sialic acid, mannosamine, glucuronic acid, galacturonic acid, mannuronic acid or guluronic acid moieties. In some embodiments, sugar is attached to protein (e.g., N- connected glycans or O- connected glycans). Exemplary sugars include glucose, galactose, fructose, mannose, rhamnose, sucrose, ribose, xylose, sialic acid, maltose, amylose, inulin, oligofructose, oligogalactose, mannan, lectin, pectin, starch, cellulose, heparin, hyaluronic acid, chitin, amylopectin or glycogen. In some embodiments, the therapeutic agent is a sugar alcohol.

[0474] In some embodiments, therapeutic agent is lipid.Lipid can be hydrophobic or amphipathic, and can form tertiary structure (such as liposome, vesicle or film) or insert in liposome, vesicle or film.Lipid can comprise fatty acid, glycerolipid, glycerophospholipid, sterol lipid, isopentenol lipid, sphingolipid, glycolipid, polyketide or sphingolipid.The example of the lipid produced by cell described herein comprises anandamide, docosahexaenoic acid, prostaglandin, leukotriene, thromboxane, eicosanoid, triglyceride, cannabinoid, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidic acid, ceramide, sphingomyelin, cerebroside, ganglioside, estrogen, androsterone, testosterone, cholesterol, carotenoid, quinone, hydroquinone or ubiquinone.

[0475] In some embodiments, therapeutic agent is a small molecule. Small molecules may include natural products produced by cells. In some embodiments, small molecule availability is poor or does not meet the Lipinski five rules (a group of guidelines for estimating whether small molecules may become human oral active drugs; See, for example, Lipinski, CA et al. (2001) Adv Drug Deliv46:2-36). Exemplary small molecule natural products include antibacterial drugs (e.g., carumonam, daptomycin, fidaxomicin, fosfomycin, ipamycin, nicardinomycin sulfate, mikamycin, mopicin, netilmicin sulfate, teicoplanin, thiomycin, rifamycin, erythromycin, vancomycin), antiparasitic drugs (e.g., artemisinin, ivermectin), anticancer drugs (e.g., doxorubicin, aclarubicin, aminolevulinic acid, arglabin, homoharringtonine, paclitaxel, pentostatin, peplomycin, romidepsin , trabectedin, actinomycin D, bleomycin, chromomycin A, daunorubicin, folinic acid, neocarcinomatine, streptozotocin, trabectedin, vinblastine, vincristine), antidiabetic drugs (for example, voglibose), central nervous system drugs (for example, levodopa, galantamine, zicontide (zicontide)), statins (for example, mevastatin), antifungal drugs (for example, fumagillin, cyclosporin), 1-deoxynojirimycin and theophylline, sterol (cholesterol, estrogen, testosterone).Other small molecule natural products are described in Newman, DJ And and Cragg, M. (2016) J Nat Prod 79:629-661 and Butler, MS et al. (2014) Nat Prod Rep 31:1612-1661.

[0476] In some embodiments, the encapsulated cells are engineered to synthesize non-protein or non-peptide small molecules. For example, in one embodiment, the cells can produce statins (e.g., taurostatin, pravastatin, fluvastatin, or atorvastatin).

[0477] In some embodiments, the therapeutic agent is an antigen (e.g., a viral antigen, a bacterial antigen, a fungal antigen, a plant antigen, an environmental antigen, or a tumor antigen). Antigens are considered by those skilled in the art to have immunostimulatory effects, i.e., to be able to stimulate an immune response or provide effective immunity to the organism or molecule from which they are derived. Antigens can be nucleic acids, peptides, proteins, sugars, lipids, or combinations thereof.

[0478] Encapsulated cells (e.g., engineered or differentiated cells, such as those described herein) can produce a single therapeutic agent or a plurality of therapeutic agents. In some embodiments, the cells produce a single therapeutic agent. In some embodiments, the hydrogel capsule encapsulates a cell cluster comprising cells that produce a single therapeutic agent. In some embodiments, at least about 1%, or about 5%, 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the cells in the cluster produce a single therapeutic agent (e.g., a therapeutic agent described herein). In some embodiments, the encapsulated cells produce a plurality of therapeutic agents, such as at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 therapeutic agents. In some embodiments, the encapsulated cell cluster comprises cells that produce a plurality of therapeutic agents. In some embodiments, at least about 1%, or about 5%, 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or 99% of the cells in the cluster produce a plurality of therapeutic agents (e.g., therapeutic agents described herein). Therapeutic agents may be related or may form a complex. In some embodiments, the therapeutic agent is secreted or released from the cell in an active form. In some embodiments, the therapeutic agent is secreted or released from the cell in an inactive form (e.g., as a prodrug). In the latter case, the therapeutic agent can be activated by a downstream agent (such as an enzyme). In some embodiments, the therapeutic agent is not secreted or released from the encapsulated cell, but is maintained in the cell. For example, the therapeutic agent can be an enzyme that participates in the detoxification or metabolism of an unwanted substance, and the detoxification or metabolism of an unwanted substance occurs in the cell.

[0479] In some embodiments, the hydrogel capsules described herein contain a concentration of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, or 50 M mL -1 In some embodiments, the hydrogel capsules described herein contain a concentration of about 1 M mL -1 In some embodiments, the hydrogel capsules described herein comprise a concentration of about 2 M mL -1 In some embodiments, the hydrogel capsules described herein contain a concentration of about 3 M mL -1 In some embodiments, the hydrogel capsules described herein contain a concentration of about 4 M mL -1 In some embodiments, the hydrogel capsules described herein contain a concentration of about 5 M mL -1 In some embodiments, the hydrogel capsules described herein comprise a concentration of about 6 M mL -1In some embodiments, the hydrogel capsules described herein comprise a concentration of about 7 M mL -1 In some embodiments, the hydrogel capsules described herein contain a concentration of about 8 MmL -1 In some embodiments, the hydrogel capsules described herein contain a concentration of about 9 M mL -1 In some embodiments, the hydrogel capsules described herein contain a concentration of about 10 M mL -1 In some embodiments, the hydrogel capsules described herein contain a concentration of about 15 M mL -1 In some embodiments, the hydrogel capsules described herein contain a concentration of about 20 M mL -1 In some embodiments, the hydrogel capsules described herein contain a concentration of about 25 M mL -1 In some embodiments, the hydrogel capsules described herein contain a concentration of about 30 M mL -1 In some embodiments, the hydrogel capsules described herein contain a concentration of about 35 M mL -1 In some embodiments, the hydrogel capsules described herein contain a concentration of about 40 M mL -1 In some embodiments, the hydrogel capsules described herein contain a concentration of about 45 M mL -1 In some embodiments, the hydrogel capsules described herein contain a concentration of about 50 M mL -1 of mammalian cells.

[0480] In some embodiments, the hydrogel capsules described herein contain a concentration of about 1-50 M mL -1 、1-45 MmL -1 、1-40 M mL -1 、1-35 M mL -1 、1-30 M mL -1 、1-25 M mL -1 、1-20 M mL -1 、1-15 M mL -1 、1-10M mL -1 , 1-5 M mL -1 , 5-50 M mL -1 、5-45 M mL -1 、5-40 M mL -1, 5-35 M mL -1 、5-30 M mL -1 、5-25M mL -1 , 5-20 M mL -1 , 5-15 M mL -1 , 5-10 M mL -1 、10-50 M mL -1 、10-45 M mL -1 、10-40 M mL -1 、10-35 M mL -1 、10-30 M mL -1 、10-25 M mL -1 、10-20 M mL -1 、10-15 M mL -1 、15-50 M mL -1 、15-45 M mL -1 、15-40 M mL -1 、15-35 M mL -1 、15-30 M mL -1 、15-25 M mL -1 、15-20 M mL -1 、20-50M mL -1 、20-45 M mL -1 、20-40 M mL -1 , 20-35 M mL -1 , 20-30 M mL -1 , or 20-25 M mL -1 between mammalian cells.

[0481] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising: a blend of VLVG alginate and SLG100 alginate, wherein the VLVG alginate comprises Compound 101 and the SLG100 alginate is unmodified; and (ii) a concentration of 5-25 M mL -1 and (ii) an outer compartment comprising a blend of VLVG alginate and SLG100 alginate, wherein the VLVG alginate comprises Compound 101 and the SLG100 alginate is unmodified.

[0482] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising: a blend of VLVG alginate and SLG100 alginate, wherein the VLVG alginate comprises Compound 101 and the SLG100 alginate comprises Compound 205; and (ii) a concentration of 5-25 M mL-1 and (ii) an outer compartment comprising a blend of VLVG alginate and SLG100 alginate, wherein the VLVG alginate comprises Compound 101 and the SLG100 alginate comprises Compound 205.

[0483] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG100 alginate and a concentration of 5-25 M mL -1 and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 205.

[0484] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified SLG20 alginate and a concentration of 5-25 M mL -1 and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 205.

[0485] In some embodiments, the hydrogel capsule comprises: (i) an inner compartment comprising unmodified LG20 alginate and a concentration of 5-25 M mL -1 and (ii) an outer compartment comprising LG20 comprising Compound 101 and Compound 205.

[0486] Treatment

[0487] Described herein are methods for preventing or treating a disease, condition, or illness in a subject by administering or implanting a hydrogel capsule comprising a polysaccharide polymer described herein. In some embodiments, the methods described herein directly or indirectly reduce or alleviate at least one symptom of the disease, condition, or illness. In some embodiments, the methods described herein prevent or slow the onset of the disease, condition, or illness. In some embodiments, the subject is a human.

[0488] In some embodiments, the disease, disorder, or condition affects a body system, such as the nervous system (e.g., peripheral or central nervous system), vascular system, skeletal system, respiratory system, endocrine system, lymphatic system, reproductive system, or gastrointestinal tract. In some embodiments, the disease, disorder, or condition affects a part of the body, such as the blood, eyes, brain, skin, lungs, stomach, mouth, ears, legs, feet, hands, liver, heart, kidneys, bones, pancreas, spleen, large intestine, small intestine, spinal cord, muscle, ovaries, uterus, vagina, or penis.

[0489] In some embodiments, the disease, disorder, or condition is a neurodegenerative disease, diabetes (type 1 or type 2), heart disease, an autoimmune disease, cancer, liver disease, a lysosomal storage disease, a blood coagulation disorder or coagulation disorder, an orthopedic disorder, or an amino acid metabolism disorder.

[0490] In some embodiments, the disease, disorder or illness is an autoimmune disease. In some embodiments, the disease, disorder or illness is diabetes (type 1 or type 2 diabetes). In one embodiment, the subject suffers from or is diagnosed with diabetes (e.g., type 1 or type 2 diabetes). The subject can have any biomarker or other diagnostic criteria associated with diabetes, such as high blood sugar levels (e.g., greater than 300 mg / dL, greater than 400 mg / dL) or high hemoglobin A1C levels (e.g., greater than 5.9% hemoglobin A1C levels, greater than 6.5% hemoglobin A1C levels, greater than 7% hemoglobin A1C levels).

[0491] In some embodiments, the disease, disorder, or condition is not type I diabetes and / or is not type II diabetes.

[0492] In some embodiments, the disease, disorder or condition is a neurodegenerative disease. Exemplary neurodegenerative diseases include Alzheimer's disease, Huntington's disease, Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS) and cerebral palsy (CP), dentatorubral-pallidular Lewy body atrophy (DRPLA), neuronal intranuclear hyaline inclusion disease (NIHID), Lewy body dementia, Down syndrome, Hallervorden-Spatz disease, prion disease, argyrophilic grain dementia, corticobasal degeneration, dementia pugilistica, diffuse neurofibrillary tangles, Gerstmann-Strauss-Scheinker disease, Jacob-Kreutzfeldt disease, Niemann-Pick disease type 3, progressive supranuclear palsy, subacute sclerosing panencephalitis, spinocerebellar ataxia, Pick's disease and dentatorubral-pallidular Lewy body atrophy.

[0493] In some embodiments, the disease, disorder, or condition is an autoimmune disease, such as scleroderma, multiple sclerosis, lupus, or allergy.

[0494] In some embodiments, the disease is a liver disease, eg, hepatitis B, hepatitis C, cirrhosis, NASH.

[0495] In some embodiments, the disease, disorder or condition is cancer. Exemplary cancers include leukemia, lymphoma, melanoma, lung cancer, brain cancer (e.g., glioblastoma), sarcoma, pancreatic cancer, kidney cancer, liver cancer, testicular cancer, prostate cancer, or uterine cancer.

[0496] In some embodiments, the disease, disorder, or condition is an orthopedic condition. Exemplary orthopedic conditions include osteoporosis, osteonecrosis, Paget's disease, or a bone fracture.

[0497] In some embodiments, the disease, disorder or condition is a lysosomal storage disease. Exemplary lysosomal storage diseases include Gaucher disease (e.g., type I, type II, type III), Tay-Sachs disease, Fabry disease, Farber disease, Hurler syndrome (also known as mucopolysaccharidosis type I (MPS I)), Hunter syndrome, lysosomal acid lipase deficiency, Niemann-Pick disease, sialidosis, Sanfilippo syndrome (also known as mucopolysaccharidosis type IIIA (MPS3A)), multiple sulfatase deficiency, Mara-Lagnes syndrome, metachromatic leukodystrophy, Krabbe disease, Scheherazade syndrome, Hurler-Schweinberg syndrome, Sly syndrome, hyaluronidase deficiency, Pompe disease, Danon disease, gangliosidosis, or Morquio syndrome.

[0498] In some embodiments, the disease, disorder or condition is a blood coagulation disorder or coagulation disorder. Exemplary blood coagulation disorders or coagulation disorders include hemophilia (e.g., hemophilia A or hemophilia B), von Willebrand disease, thrombocytopenia, uremia, Bernard-Soulier syndrome, factor XII deficiency, vitamin K deficiency, or afibrinogenemia.

[0499] In some embodiments, the disease, condition, or disorder is an amino acid metabolism disorder, such as phenylketonuria, tyrosinemia (eg, type 1 or type 2), alkaptonuria, homocystinuria, hyperhomocysteinemia, maple syrup urine disease.

[0500] In some embodiments, the disease, disorder, or condition is a fatty acid metabolism disorder, such as hyperlipidemia, hypercholesterolemia, galactosemia.

[0501] In some embodiments, the disease, disorder, or condition is a disorder of purine or pyrimidine metabolism, such as Lesch-Nyhan syndrome.

[0502] The present invention further includes methods for identifying a subject having or suspected of having a disease, condition, or illness as described herein, and after such identification, administering to the subject an implantable element comprising cells (e.g., optionally encapsulated by a closure member and optionally modified with a compound of formula (I) or a composition thereof as described herein). In one embodiment, the subject is human. In one embodiment, the subject is human. In one embodiment, the subject is an adult. In one embodiment, the subject is a child (e.g., a subject less than 21 years of age, less than 18 years of age, less than 15 years of age, less than 12 years of age, less than 10 years of age, or less than 6 years of age).

[0503] Method for making particles

[0504] The present disclosure further includes methods for making particles described herein, e.g., particles comprising a polysaccharide polymer, comprising a first compartment, a second compartment, a photoactive cross-linking moiety, and a compound of formula (I). In some embodiments where the particles are hydrogel capsules, the method for making the hydrogel capsules comprises contacting a plurality of droplets comprising a first and a second polymer solution (e.g., each comprising a hydrogel-forming polymer) with an aqueous cross-linking solution. In one embodiment, the aqueous cross-linking solution comprises an ionic cross-linking agent, e.g., a divalent cation such as calcium, barium, and magnesium. The droplets can be formed using any technique known in the art. In further embodiments where the particles are hydrogel capsules, the method for making the hydrogel capsules comprises an irradiation step, wherein the solution comprising the polysaccharide polymer is exposed to ultraviolet light to initiate a photocross-linking reaction. In one embodiment, one or both of the first and second polymer solutions may further contain a photoinitiator.

[0505] Each compartment of the particles described herein may comprise any one or more of the following: an unmodified polymer, a polymer modified with one or both of a compound of formula (I) and a photoactive cross-linking moiety, or a blend of an unmodified polymer and a modified polymer. In brief, in performing a method for preparing particles constructed as two-compartment hydrogel capsules, a volume of a first polymer solution (e.g., comprising an unmodified polymer, a polymer modified with a compound of formula (I), a photoactive cross-linking moiety, or a blend thereof, and optionally containing cells) is loaded into a first syringe connected to the lumen of a coaxial needle. The first syringe can then be connected to a syringe pump vertically oriented above a container containing an aqueous cross-linking solution comprising a cross-linking agent, a buffer, and an osmotic concentration regulator. A volume of a second polymer solution (e.g., comprising an unmodified polymer, a polymer modified with a compound of formula (I), a photoactive cross-linking moiety, or a blend thereof, and optionally containing cells) is loaded into a second syringe connected to the outer lumen of a coaxial needle. The second syringe can then be connected to a syringe pump horizontally oriented relative to the container containing the cross-linking solution. The high-voltage power generator can then be connected to the top and bottom of pin. Syringe pump and power generator can then be used to extrude the first and second polymer solutions by syringe, and the setting of this syringe is determined to reach the required drop rate that polymer solution enters cross-linking solution. Those skilled in the art can easily determine the various combinations of pin lumen size, voltage range, flow rate, drop rate and drip distance to produce 2 compartment hydrogel capsule compositions, wherein most of (for example at least 80%, 85%, 90% or more) capsules are in 10% of target size and have class spheroid shape. After exhausting the polymer solution of the first and second volumes, drop can be allowed to crosslink a certain amount of time in cross-linking solution, for example, approximately five minutes. Cross-linking may require ionic crosslinking (for example, by making drop and ionic crosslinking agent, for example divalent cation contact) and / or covalent crosslinking (for example, by irradiating drop with activation photoactive crosslinking agent, for example methacrylate or methacrylamide).

[0506] Exemplary process parameters for preparing millimeter-sized capsules (e.g., 1.5 mm diameter millimeter-sized capsules) compositions include the following. A coaxial needle is positioned above the surface of the cross-linking solution at a distance sufficient to provide a droplet distance from the needle tip to the surface of the solution. In one embodiment, the distance between the needle tip and the surface of the solution is between 1 cm and 5 cm. In one embodiment, the first and second polymer solutions are extruded through the needle at a total flow rate of 0.05 mL / min to 5 mL / min, or 0.05 mL / min to 2.5 mL / min, or 0.05 mL / min to about 1 mL / min, or 0.05 mL / min to 0.5 mL / min, or 0.1 mL / min to 0.5 mL / min. In one embodiment, the first and second polymer solutions are extruded through the needle at a total flow rate of approximately 0.05 mL / min, 0.1 mL / min, 0.15 mL / min, 0.2 mL / min, 0.25 mL / min, 0.3 mL / min, 0.35 mL / min, 0.4 mL / min, 0.45 mL / min, or 0.5 mL / min. In one embodiment, the flow rates of the first and second polymer solutions through the needle are substantially the same. In one embodiment, the flow rates of the first and second polymer solutions through the needle are different.

[0507] In one embodiment, the voltage of the instrument is between 1 kV and 20 kV, or between 1 kV and 15 kV, or between 1 kV and 10 kV, or between 5 kV and 10 kV. The voltage can be adjusted until the desired drop rate is achieved. In one embodiment, the drop rate of the instrument is between 1 drop / 10 seconds and 50 drops / 10 seconds, or between 1 drop / 10 seconds and 25 drops / 10 seconds.

[0508] In one embodiment, the number of non-particle debris on the surface of the cross-linking solution is determined. The particles that fall to the bottom of the cross-linking container can then be collected, for example, by transferring the cross-linking solution containing the particles to a separate container, leaving any non-particle debris on the surface of the solution in the original cross-linking container. The removed particles can then be allowed to settle, the cross-linking solution can be removed, and the particles can then be washed one or more times with a buffer (e.g., HEPES buffer). In one embodiment, one or more aliquots of the resulting particle composition (e.g., particle preparation) are examined by microscopy to assess the quality of the composition, such as the number of particle defects and satellite particles.

[0509] In some embodiments, the cross-linking solution further comprises a process additive (e.g., a hydrophilic nonionic surfactant). The process additive can reduce the surface tension of the cross-linking solution. Reagents that can be used as process additives in the present disclosure include polysorbate-type surfactants, copolymers of polyethylene oxide (PEO) and polypropylene oxide (PPO), poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) triblock copolymers, and nonionic surfactants such as Tween ® 20. Tween ® 80, Triton™ X-100, IGEPAL® CA-630, poloxamer 188, or poloxamer 407, or a surfactant having substantially the same chemical and physical properties as listed in the table of exemplary surfactants immediately below.

[0510]

[0511] a Hydrophile-lipophile balance

[0512] b Chemical name and synonyms: Polyethylene glycol sorbitan monolaurate, Polyoxyethylene (20) sorbitan monolaurate, Polysorbate 20, Polyoxyethylene 20 sorbitan monododecanoate

[0513] c Chemical name and synonyms: Polyethylene glycol sorbitan monooleate, polyoxyethylene (20) sorbitan monooleate, polysorbate 80, (x)-sorbitan mono-9-octadecenoate poly(oxy-1,2-ethanediyl)

[0514] d Chemical name and synonyms: 4-(1,1,3,3-tetramethylbutyl)phenyl-polyethylene glycol, tert-octylphenoxypolyethoxyethanol, polyethylene glycol tert-octylphenyl ether; octylphenol ethoxylate, octylphenol ethylene oxide condensate

[0515] e Chemical name and synonyms: Branched octylphenoxypolyethoxyethanol, octylphenoxypoly(ethyleneoxy)ethanol

[0516] f Chemical name: Poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol)

[0517] g Chemical name: Poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol)

[0518] In some embodiments, the process additive is a nonionic surfactant. In one embodiment, the process additive comprises more than one surfactant, for example, more than one hydrophilic surfactant. In some embodiments, the process additive does not contain Tween® 20 (polysorbate 20) or Triton™ X-100. In one embodiment, the process additive is IGEPAL® CA-630 (polyethylene glycol sorbitan monooleate). In some embodiments, the process additive is poloxamer 188.

[0519] In some embodiments, a process additive (e.g., a surfactant) is present in the cross-linking solution at a concentration of at least 0.0001% or higher. In some embodiments, the cross-linking solution comprises at least 0.001%, 0.01%, or 0.1% of a process additive. In some embodiments, the process additive is present at a concentration selected from about 0.001% to about 0.1%, about 0.005% to about 0.05%, about 0.005% to about 0.01%, and about 0.01% to about 0.5%. In one embodiment, the process additive is a surfactant and is present at a concentration below the critical micelle concentration of the surfactant.

[0520] In some embodiments, the ionic crosslinker comprises a single type of divalent cation or a mixture of different types, such as Ba 2+ , Ca 2+ 、Sr 2+ In some embodiments, the ionic crosslinking agent is, for example, BaCl2 at a concentration of 1 mM to 100 mM or 7.5 mM to 20 mM. In some embodiments, the ionic crosslinking agent is, for example, CaCl2 at a concentration of 50 mM to 100 mM. In some embodiments, the ionic crosslinking agent is, for example, SrCl2 at a concentration of 37.5 mM to 100 mM. In some embodiments, the ionic crosslinking agent is a mixture of BaCl2 (e.g., 5 mM to 20 mM) and CaCl2 (e.g., 37.5 mM to 12.5 mM) or a mixture of BaCl2 (e.g., 5 mM to 20 mM) and SrCl2 (e.g., 37.5 mM to 12.5 mM).

[0521] In some embodiments, the ionic crosslinker is SrCl2 and the process additive is Tween ®80 (or a surfactant having substantially the same chemical and physical properties as listed in the exemplary surfactant table) at a concentration of less than 0.1%, for example, from about 0.005% to 0.05%, from about 0.005% to about 0.01%. In some embodiments, the concentration of SrCl2 is about 50 mM. In some embodiments, the ionic crosslinker is SrCl2 and the process additive is poloxamer 188 at a concentration of 1%.

[0522] The type and concentration of the buffer in the aqueous cross-linking solution are selected to maintain the solution pH at approximately neutral, e.g., from about 6.5 to about 7.5, from about 7.0 to about 7.5, or about 7.0. In one embodiment, the buffer is compatible with the biological material (e.g., cells) to be encapsulated in the particles. In some embodiments, the buffer in the aqueous cross-linking solution comprises HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid).

[0523] The osmotic pressure regulating agent in the aqueous cross-linking solution is selected to maintain the solution osmotic pressure at a value similar to the osmotic pressure of the polymer solution (which, in some embodiments, includes a cell suspension), for example, with a variation of up to 20%, 10%, or 5% higher or lower osmotic pressure. In some embodiments, the osmotic pressure regulating agent is mannitol at a concentration of 0.1 M to 0.3 M.

[0524] In some embodiments, the cross-linking solution comprises 25 mM HEPES buffer, 20 mM BaCl2, 0.2 M mannitol, and 0.01% poloxamer 188.

[0525] In some embodiments, the cross-linking solution comprises 50 mM strontium chloride hexahydrate, 0.165 M mannitol, 25 mM HEPES, and 0.01% of a surfactant having substantially the same chemical and physical properties as listed in the exemplary surfactant table for Tween 80.

[0526] In one embodiment, the process additive is poloxamer 188, which is present in a detectable amount in the particle composition (e.g., particle formulation) after the washing step. Poloxamer 188 can be detected by any technique known in the art, such as by partially or completely dissolving the particles in an aliquot of the composition via sodium sulfate precipitation and analyzing the supernatant by LC / MS.

[0527] The reduction in surface tension of the cross-linking solution can be assessed by any method known in the art, such as by using a contact angle goniometer or a tensiometer, for example via the du Nouy ring method (see, e.g., Davarci et al. (2017) Food Hydrocolloids 62: 119-127).

[0528] Exemplary embodiments

[0529] 1. A polysaccharide polymer comprising:

[0530] (i) a photoactive cross-linking moiety; and

[0531] (ii) Compound of formula (I):

[0532] (I),

[0533] or a pharmaceutically acceptable salt thereof, wherein:

[0534] A is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -O-, -C(O)O-, -C(O)-, -OC(O)-, -N(R C )-、-N(R C )C(O)-、-C(O)N(R C )-、-N(R C )N(R D )-、-NCN-、-N(R C )C(O)(C1-C6-alkylene)-, -N(R C )C(O)(C2-C6-alkenylene)-、-C(=N(R C )(R D ))O-、-S-、-S(O) x -、-OS(O) x -、-N(R C )S(O) x -、-S(O) x N(R C )-、-P(R F ) y -、-Si(OR A )2-、-Si(R G )(OR A )-、-B(OR A )- or metal, each of which is optionally linked to an attachment group (e.g., an attachment group described herein) and optionally surrounded by one or more R 1 replace;

[0535] L 1 and L 3 Each of the R 2 replace;

[0536] L 2 It is a key;

[0537] M is absent, alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is optionally replaced by one or more R 3 replace;

[0538] P is optionally replaced by one or more R 4 substituted heteroaryl;

[0539] Z is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally substituted by one or more R 5 replace;

[0540] Each R A 、R B 、R C 、R D 、R E 、R F and R G are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, azido, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally replaced by one or more R 6 replace;

[0541] or R C and R D Together with the nitrogen atom to which they are attached, they form a 6 substituted rings (e.g., 5-7 membered rings);

[0542] Each R 1 、R 2 、R 3 、R 4 、R 5 and R 6 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 、–C(O)N(R C1 ), SR E1 、S(O) x R E1 、–OS(O) x R E1 ,–N(R C1 )S(O) x RE1 ,–S(O) x N(R C1 )(R D1 ),–P(R F1 ) y , cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally replaced by one or more R 7 replace;

[0543] Each R A1 、R B1 、R C1 、R D1 、R E1 and R F1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally replaced by one or more R 7 replace;

[0544] Each R 7 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, oxo, hydroxy, cycloalkyl, or heterocyclyl;

[0545] x is 1 or 2; and

[0546] y is 2, 3, or 4.

[0547] 2. The polysaccharide polymer of embodiment 1, wherein the photoactive cross-linking moiety is covalently bound to a sugar monomer within the polysaccharide polymer.

[0548] 3. The polysaccharide polymer of embodiment 2, wherein the photoactive cross-linking moiety is bound to a carboxylate moiety within the sugar monomer.

[0549] 4. The polysaccharide polymer of any one of embodiments 1 to 3, wherein the photoactive cross-linking moiety comprises an alkyl, alkenyl, alkynyl, ester, ketone, amine, or amide group.

[0550] 5. The polysaccharide polymer of any one of embodiments 1 to 3, wherein the photoactive cross-linking moiety comprises an alkyl group.

[0551] 6. The polysaccharide polymer of any one of embodiments 1 to 3, wherein the photoactive cross-linking moiety comprises an alkenyl group.

[0552] 7. The polysaccharide polymer of any one of embodiments 1 to 3, wherein the photoactive cross-linking moiety comprises an alkyne group.

[0553] 8. The polysaccharide polymer of any one of embodiments 1 to 3, wherein the photoactive cross-linking moiety comprises an ester group.

[0554] 9. The polysaccharide polymer of any one of embodiments 1 to 3, wherein the photoactive cross-linking moiety comprises a keto group.

[0555] 10. The polysaccharide polymer of any one of embodiments 1 to 3, wherein the photoactive cross-linking moiety comprises an amine group.

[0556] 11. The polysaccharide polymer of any one of embodiments 1 to 3, wherein the photoactive cross-linking moiety comprises an amide group.

[0557] 12. The polysaccharide polymer of any one of embodiments 1-11, wherein the photoactive cross-linking moiety is capable of reacting with a second photoactive cross-linking moiety when activated by light (eg, ultraviolet light).

[0558] 13. The polysaccharide polymer of any one of embodiments 1-12, wherein the photoactive cross-linking moieties are present on the polysaccharide polymer at a density of at least about 1%, e.g., 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or more, e.g., as determined by comparison to a reference standard (e.g., as determined by LC-MS assay).

[0559] 14. The polysaccharide polymer of any one of embodiments 1-13, wherein the photoactive cross-linking moieties are present on the polysaccharide polymer at a density between 1% and 10%, e.g., 1% to 8%, 1% to 6%, or 1% to 4%, e.g., as determined by comparison to a reference standard.

[0560] 15. The polysaccharide polymer of any one of embodiments 1-14, wherein the photoactive cross-linking moieties are present on the polysaccharide polymer at a density between 1% and 8%, eg, as determined by comparison to a reference standard.

[0561] 16. The polysaccharide polymer of any one of embodiments 1-15, wherein the photoactive cross-linking moieties are present on the polysaccharide polymer at a density between 1% and 6%, eg, as determined by comparison to a reference standard.

[0562] 17. The polysaccharide polymer of any one of embodiments 1-16, wherein the photoactive cross-linking moieties are present on the polysaccharide polymer at a density between 1% and 4%, eg, as determined by comparison to a reference standard.

[0563] 18. The polysaccharide polymer of any one of embodiments 1-17, wherein the polysaccharide polymer is selected from the group consisting of agarose, alginate, amylose, amylopectin, arabinogalactan, cellulose, chitin, chitosan, dextran, fructan, fucoidan, galactan, galactomannan, glycogen, gellan gum, hyaluronic acid, hyaluronate, inulin, laminarin, maltodextrin, pectin, pullulan, xanthan gum, xylan, carrageenan, and raffinose.

[0564] 19. The polysaccharide polymer of any one of embodiments 1-18, wherein the polysaccharide polymer is not agarose, amylose, amylopectin, arabinogalactan, cellulose, chitin, chitosan, dextran, fructan, fucoidan, galactan, galactomannan, glycogen, gellan gum, hyaluronic acid, hyaluronate, inulin, laminarin, maltodextrin, pectin, polytriglucose, xanthan gum, xylan, carrageenan, and raffinose.

[0565] 20. The polysaccharide polymer of any one of embodiments 1-19, wherein the polysaccharide polymer is selected from the group consisting of alginate, hyaluronate, and chitosan.

[0566] 21. The polysaccharide polymer of any one of embodiments 1-20, wherein the polysaccharide polymer is alginate.

[0567] 22. The polysaccharide polymer of any one of embodiments 1-21, wherein the polysaccharide polymer is hyaluronate.

[0568] 23. The polysaccharide polymer of any one of embodiments 1-22, wherein the polysaccharide polymer is chitosan.

[0569] 24. The polysaccharide polymer of embodiment 21, wherein the alginate is high guluronic acid (G) alginate or high mannuronic acid (M) alginate.

[0570] 25. The polysaccharide polymer of embodiment 21 or 24, wherein the alginate is homoguluronic acid (G) alginate.

[0571] 26. The polysaccharide polymer of embodiments 21 or 24-25, wherein the alginate is high mannuronic acid (G) alginate.

[0572] 27. The polysaccharide polymer of any one of embodiments 21 or 24-26, wherein the alginate is not high mannuronic acid (M) alginate.

[0573] 28. The polysaccharide polymer of any one of embodiments 21 or 24-27, wherein the alginate is selected from the group consisting of low molecular weight alginate, medium molecular weight alginate, high molecular weight alginate, and ultra-high molecular weight alginate.

[0574] 29. The polysaccharide polymer of any one of embodiments 21 or 24-28, wherein the alginate is a low molecular weight alginate.

[0575] 30. The polysaccharide polymer of any one of embodiments 21 or 24-29, wherein the alginate is a medium molecular weight alginate.

[0576] 31. The polysaccharide polymer of any one of embodiments 21 or 24-30, wherein the alginate is a high molecular weight alginate.

[0577] 32. The polysaccharide polymer of any one of embodiments 21 or 24-31, wherein the alginate is an ultra-high molecular weight alginate.

[0578] 33. The polysaccharide polymer of any one of embodiments 21 or 24-32, wherein the alginate is selected from the group consisting of low viscosity alginate, medium viscosity alginate, high viscosity alginate, and very high viscosity alginate.

[0579] 34. The polysaccharide polymer of any one of embodiments 21 or 24-33, wherein the alginate is a low viscosity alginate.

[0580] 35. The polysaccharide polymer of any one of embodiments 21 or 24-34, wherein the alginate is a medium viscosity alginate.

[0581] 36. The polysaccharide polymer of any one of embodiments 21 or 24-35, wherein the alginate is a high viscosity alginate.

[0582] 37. The polysaccharide polymer of any one of embodiments 21 or 24-36, wherein the alginate is a very high viscosity alginate.

[0583] 38. The polysaccharide polymer of any one of embodiments 21 or 24-37, wherein the alginate has a purity greater than 90%, e.g., greater than 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% (e.g., as determined by high performance liquid chromatography).

[0584] 39. The polysaccharide polymer of any one of embodiments 21 or 24-38, wherein the alginate has a purity greater than 95%.

[0585] 40. The polysaccharide polymer of any one of embodiments 21 or 24-39, wherein the alginate has a purity greater than 98%.

[0586] 41. The polysaccharide polymer of any one of embodiments 21 or 24-40, wherein the alginate has a purity greater than 99%.

[0587] 42. The polysaccharide polymer of any one of embodiments 21 or 24-41, wherein the alginate has a purity greater than 99.5%.

[0588] 43. The polysaccharide polymer of any one of embodiments 21 or 24-42, wherein the alginate has an endotoxin level of less than 50 EU / g.

[0589] 44. The polysaccharide polymer of any one of embodiments 21 or 24-43, wherein the alginate has a total viable bacterial count (TVC) of less than 100 colony forming units (CFU) per gram.

[0590] 45. A polysaccharide polymer as described in any one of embodiments 21 or 24-44, wherein the alginate is selected from one of the following: Kimica Algin IL-2, Kimica Algin IL-6, Kimica Algin 1-1, Kimica Algin 1-3, Kimica Algin 1-5, Kimica Algin 1-8, Kimica Algin LZ-2, Kimica Algin ULV-L3, Kimica Algin ULV-L5, Kimica Algin ULV-1G, Kimica Algin ULV-5G, Kimica Algin ULVIL-6G, Pronova UP VLVM, Pronova UP LVM, Pronova UP MVM, Pronova UP VLVG, Pronova UP MVG, Pronova UP LVG, Pronova SLM20, Pronova SLM100, Pronova SLG20, Pronova UPLG 20, Pronova LG 100 and Pronova SLG100.

[0591] 46. ​​The polysaccharide polymer of any one of embodiments 21 or 24-45, wherein the alginate is selected from one of the following: Pronova UP VLVM, Pronova UP LVM, Pronova UP MVM, Pronova UP VLVG, Pronova UP MVG, Pronova UP LVG, Pronova SLM20, Pronova SLM100, Pronova SLG20, and Pronova SLG100.

[0592] 47. The polysaccharide polymer of any one of embodiments 21 or 24-46, wherein the alginate is Pronova UP VLVG.

[0593] 48. The polysaccharide polymer of any one of embodiments 21 or 24-46, wherein the alginate is Pronova SLG100.

[0594] 49. The polysaccharide polymer of any one of embodiments 21 or 24-46, wherein the alginate is Pronova SLG20.

[0595] 50. The polysaccharide polymer of embodiment X, wherein the alginate is Pronova LG 20.

[0596] 51. The polysaccharide polymer of any one of embodiments 1-50, wherein the photoactive cross-linking moiety has formula (IV):

[0597] (IV), or a pharmaceutically acceptable salt or tautomer thereof, wherein:

[0598] X 1 Does not exist, O, NR 33 or C(R 34a )(R 34b );

[0599] R 30a 、R 30b 、R 31 、R 32 、R 33 、R 34a and R 34b Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(RD1 ),–N(R C1 )C(O)R B1 ,–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl;

[0600] Each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 replace; and

[0601] Each R 7 is independently alkyl, alkenyl, alkynyl, heteroalkyl, halo, cyano, oxo, hydroxy, cycloalkyl, or heterocyclyl.

[0602] 52. The polysaccharide polymer of embodiment 51, wherein X 1 NR 33 (e.g. NH).

[0603] 53. A polysaccharide polymer as described in any one of embodiments 51-52, wherein R 30a 、R 30b 、R 31 and R 32 Each of is hydrogen.

[0604] 54. The polysaccharide polymer of embodiment 51, wherein X 1 NR 33 , R 30a 、R 30b 、R 31 and R 33 Each of is hydrogen and R 32 is a heteroalkyl group (eg, a nitrogen-containing heteroalkyl group).

[0605] 55. The polysaccharide polymer of embodiment 51, wherein X 1 It is NR 33 , R 30a 、R 30b and R 33 Each of R is hydrogen, 31 is an alkyl group, and R 32 is a heteroalkyl group (eg, a nitrogen-containing heteroalkyl group).

[0606] 56. The polysaccharide polymer of embodiment 51, wherein X1 It is NR 33 , R 30a 、R 30b and R 33 Each of R is hydrogen, 31 It is C1-C 12 Alkyl, and R 32 is a 2- to 12-membered heteroalkyl group (eg, a 2- to 12-membered nitrogen-containing heteroalkyl group).

[0607] 57. The polysaccharide polymer of embodiment 48, wherein X 1 It's O, R 30a 、R 30b 、R 31 and R 33 Each of is hydrogen and R 32 is a heteroalkyl group (eg, a nitrogen-containing heteroalkyl group).

[0608] 58. The polysaccharide polymer of embodiment 51, wherein X 1 It's O, R 30a and R 30b Each of R is hydrogen, 31 is an alkyl group, and R 32 is a heteroalkyl group (eg, a nitrogen-containing heteroalkyl group).

[0609] 59. The polysaccharide polymer of embodiment 51, wherein X 1 It's O, R 30a and R 30b Each of R is hydrogen, 31 It is C1-C 12 Alkyl, and R 32 is a 2- to 12-membered heteroalkyl group (eg, a 2- to 12-membered nitrogen-containing heteroalkyl group).

[0610] 60. The polysaccharide polymer of embodiment 51, wherein X 1 It is C(R 34a )(R 34b ), R 30a 、R 30b 、R 31 、R 33 、R 34a and R 34b Each of is hydrogen and R 32 is a heteroalkyl group (eg, a nitrogen-containing heteroalkyl group).

[0611] 61. The polysaccharide polymer of embodiment 51, wherein X 1 It is C(R 34a )(R 34b ), R 30a 、R 30b 、R31 、R 33 、R 34a and R 34b Each of R is hydrogen, 31 is an alkyl group, and R 32 is a heteroalkyl group (eg, a nitrogen-containing heteroalkyl group).

[0612] 62. The polysaccharide polymer of embodiment 51, wherein X 1 It is C(R 34a )(R 34b ), R 30a 、R 30b 、R 31 、R 33 、R 34a and R 34b Each of R is hydrogen, 31 It is C1-C 12 Alkyl, and R 32 is a 2- to 12-membered heteroalkyl group (eg, a 2- to 12-membered nitrogen-containing heteroalkyl group).

[0613] 63. The polysaccharide polymer of embodiment 51, wherein X 1 Not O.

[0614] 64. The polysaccharide polymer of embodiment 51, wherein X 1 It's O, R 32 is a 2-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, or 12-membered heteroalkyl group.

[0615] 65. The polysaccharide polymer of embodiment 51, wherein X 1 It is NR 33 , R 32 is a 2-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, or 12-membered heteroalkyl group.

[0616] 66. The polysaccharide polymer of embodiment 48, wherein X 1 It is O and R 32 Not a 3-membered heteroalkyl group.

[0617] 67. The polysaccharide polymer of any one of embodiments 48-66, wherein the photoactive cross-linking moiety is selected from the group consisting of: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 and or a pharmaceutically acceptable salt thereof.

[0618] 68. The polysaccharide polymer of any one of embodiments 48-66, wherein the photoactive cross-linking moiety is selected from the group consisting of: 、 、 、 、 、 、 、 、 、 、 、 、 、 and or a pharmaceutically acceptable salt thereof.

[0619] 69. The polysaccharide polymer of any one of embodiments 48-66, wherein the photoactive cross-linking moiety is selected from the group consisting of: 、 、 、 、 、 、 and or a pharmaceutically acceptable salt thereof.

[0620] 70. The polysaccharide polymer of any one of embodiments 1-69, wherein the photoactive cross-linking moiety has formula (IV-a):

[0621] (IV-a), or a pharmaceutically acceptable salt or tautomer thereof, wherein: R 30a 、R 30b 、R 31 、R 32 and R 35 Each of the groups is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 、–C(O)OR A1 ,–C(O)R B1 、–OC(O)R B1 ,–N(R C1 )(R D1),–N(R C1 )C(O)R B1 ,–C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl;

[0622] Each R A1 、R B1 、R C1 、R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted ...

Claims

1. A polysaccharide polymer comprising: (i) a photoactive cross-linking moiety; and (ii) a compound of formula (I): (I)， or a pharmaceutically acceptable salt thereof, wherein: A is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -O-, -C(O)O-, -C(O)-, -OC(O)-, -N(R C )-、-N(R C )C(O)-、-C(O)N(R C )-、-N(R C )N(R D )-、-NCN-、-N(R C )C(O)(C1-C6-alkylene)-, -N(R C )C(O)(C2-C6-alkenylene)-, -C(=N(R C )(R D ))O-、-S-、-S(O) x -、-OS(O) x -、-N(R C )S(O) x -、-S(O) x N(R C )-、-P(R F ) y -、-Si(OR A )2-、-Si(R G )(OR A )-、-B(OR A )- or a metal, each of which is optionally linked to an attachment group (e.g., an attachment group described herein) and is optionally bound by one or more R 1 replace; L 1 and L 3 Each of is independently a bond, an alkyl group or a heteroalkyl group, wherein each alkyl group and heteroalkyl group is optionally substituted by one or more R 2 replace; L 2 is the key; M is absent, alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally replaced by one or more R 3 replace; P is optionally replaced by one or more R 4 substituted heteroaryl; Z is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally substituted by one or more R 5 replace; Each R A , R B , R C , R D , R E , R F and R G are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, azido, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally substituted by one or more R 6 replace; or R C and R D Together with the nitrogen atom to which they are attached, they form a 6 substituted rings (e.g., 5-7 membered rings); Each R 1 , R 2 , R 3 , R 4 , R 5 and R 6 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, –OR A1 , –C(O)OR A1 , –C(O)R B1 、–OC(O)R B1 , –N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 , –C(O)N(R C1 ), SR E1 、S(O) x R E1 、–OS(O) x R E1 , –N(R C1 )S(O) x R E1 , –S(O) x N(R C1 )(R D1 ),–P(R F1 ) y , cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally substituted by one or more R 7 replace; Each R A1 , R B1 , R C1 , R D1 , R E1 and R F1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by one or more R 7 replace; Each R 7 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxy, cycloalkyl or heterocyclyl; x is 1 or 2; and y is 2, 3, or 4.

2. The polysaccharide polymer of claim 1, wherein the photoactive cross-linking moiety is covalently bonded to a sugar monomer within the polysaccharide polymer.

3. The polysaccharide polymer of claim 2, wherein the photoactive cross-linking moiety is bound to a carboxylate moiety within the sugar monomer.

4. The polysaccharide polymer of any one of claims 1 to 3, wherein the photoactive cross-linking moiety comprises an alkyl, alkenyl, alkynyl, ester, ketone, amine or amide group.

5. The polysaccharide polymer of any one of claims 1 to 4, wherein the photoactive cross-linking moiety is capable of reacting with a second photoactive cross-linking moiety when activated by light (eg, UV light).

6. The polysaccharide polymer of any one of claims 1-5, wherein the photoactive cross-linked moieties are present on the polysaccharide polymer at a density of at least about 1%, e.g., 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or more, e.g., as determined by comparison with a reference standard.

7. The polysaccharide polymer of any one of claims 1-6, wherein the photoactive cross-linking moieties are present on the polysaccharide polymer at a density between 1% and 10%, such as 1% to 8%, 1% to 6%, or 1% to 4%, such as determined by comparison with a reference standard.

8. The polysaccharide polymer of any one of claims 1 to 7, wherein the polysaccharide polymer is selected from alginate, hyaluronate and chitosan.

9. The polysaccharide polymer of any one of claims 1 to 8, wherein the polysaccharide polymer is alginate.

10. The polysaccharide polymer according to claim 9, wherein the alginate is high guluronic acid (G) alginate or high mannuronic acid (M) alginate.

11. The polysaccharide polymer of any one of claims 1 to 8, wherein the photoactive cross-linking moiety has formula (IV): (IV), or a pharmaceutically acceptable salt or tautomer thereof, wherein X 1 Does not exist, O, NR 33 or C(R 34a )(R 34b ); R 30a , R 30b , R 31 , R 32 , R 33 , R 34a and R 34b Each of the above is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 , –C(O)OR A1 , –C(O)R B1 、–OC(O)R B1 , –N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 , –C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; Each R A1 , R B1 , R C1 , R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 replace; and Each R 7 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxy, cycloalkyl or heterocyclyl.

12. The polysaccharide polymer according to claim 11, wherein X 1 NR 33 (e.g. NH).

13. The polysaccharide polymer according to any one of claims 11 to 12, wherein R 30a , R 30b , R 31 and R 32 Each of is hydrogen.

14. The polysaccharide polymer of any one of claims 1 to 13, wherein the photoactive cross-linking moiety has formula (IV-a): (IV-a), or a pharmaceutically acceptable salt or tautomer thereof, wherein: R 30a , R 30b , R 31 , R 32 and R 35 Each of the above is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 , –C(O)OR A1 , –C(O)R B1 、–OC(O)R B1 , –N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 , –C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; Each R A1 , R B1 , R C1 , R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 replace; and Each R 7 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxy, cycloalkyl or heterocyclyl.

15. The polysaccharide polymer according to claim 14, wherein R 30a , R 30b , R 31 , R 32 and R 35 Each of is hydrogen.

16. The polysaccharide polymer of any one of claims 1 to 13, wherein the photoactive cross-linking moiety has formula (IV-c): (IV-c), or a pharmaceutically acceptable salt or tautomer thereof, wherein: R 30a , R 30b and R 31 Each of the above is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 , –C(O)OR A1 , –C(O)R B1 、–OC(O)R B1 , –N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 , –C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; R 32 is hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, –C(O)OR A1 , –C(O)R B1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; Each R A1 , R B1 , R C1 , R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 replace; and Each R 7 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxy, cycloalkyl or heterocyclyl.

17. The polysaccharide polymer according to claim 16, wherein R 30a , R 30b , R 31 and R 32 Each of is hydrogen.

18. The polysaccharide polymer of any one of claims 1 to 17, wherein the photoactive cross-linking moiety has a structure selected from Table 4, or a pharmaceutically acceptable salt thereof.

19. The polysaccharide polymer of any one of claims 1 to 18, wherein the photoactive cross-linking moiety is selected from acrylates, methacrylates, acrylamides and methacrylamides, or their corresponding acid chlorides and anhydrides.

20. The polysaccharide polymer of any one of claims 1 to 19, wherein the photoactive cross-linking moiety is selected from Compound 205 or Compound 217, or a pharmaceutically acceptable salt thereof.

21. The polysaccharide polymer according to any one of claims 1 to 20, wherein the compound of formula (I) has a structure selected from Table 3, or a pharmaceutically acceptable salt thereof.

22. The polysaccharide polymer according to any one of claims 1 to 21, wherein the compound of formula (I) is selected from Compound 100, Compound 101, Compound 110, Compound 112, Compound 113, Compound 114, Compound 122 and Compound 123, or a pharmaceutically acceptable salt thereof.

23. The polysaccharide polymer according to any one of claims 1 to 22, wherein the compound of formula (I) is compound 101 or a pharmaceutically acceptable salt thereof.

24. The polysaccharide polymer of any one of claims 1 to 23, wherein the polysaccharide polymer is alginate, the photoactive cross-linking moiety is selected from Compound 205 and Compound 217 or a pharmaceutically acceptable salt thereof, and the compound of formula (I) is Compound 101 or a pharmaceutically acceptable salt thereof.

25. A composition comprising the polysaccharide polymer of any one of claims 1-24.

26. A hydrogel capsule comprising the polysaccharide polymer according to any one of claims 1 to 24.

27. The hydrogel capsule of claim 26, wherein the hydrogel capsule comprises a single compartment comprising the polysaccharide polymer (eg, a polysaccharide polymer described herein).

28. The hydrogel capsule of claim 27, wherein the hydrogel capsule comprises a plurality of compartments, wherein one of the compartments comprises the polysaccharide polymer (eg, a polysaccharide polymer described herein).

29. The hydrogel capsule of claim 28, wherein the hydrogel capsule comprises an inner compartment and an outer compartment.

30. The hydrogel capsule of any one of claims 26-29, wherein: The inner compartment comprises a first polysaccharide polymer comprising a photoactive cross-linking moiety; The outer compartment comprises a second polysaccharide polymer comprising photoactive cross-linking moieties.

31. A hydrogel capsule comprising: (i) an inner compartment comprising a first polysaccharide polymer comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof: Formula (I): (I)， in: A is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -O-, -C(O)O-, -C(O)-, -OC(O)-, -N(R C )-、-N(R C )C(O)-、-C(O)N(R C )-、-N(R C )N(R D )-、-NCN-、-N(R C )C(O)(C1-C6-alkylene)-, -N(R C )C(O)(C2-C6-alkenylene)-, -C(=N(R C )(R D ))O-、-S-、-S(O) x -、-OS(O) x -、-N(R C )S(O) x -、-S(O) x N(R C )-、-P(R F ) y -、-Si(OR A )2-、-Si(R G )(OR A )-、-B(OR A )- or a metal, each of which is optionally linked to an attachment group (e.g., an attachment group described herein) and is optionally bound by one or more R 1 replace; L 1 and L 3 Each of is independently a bond, an alkyl group or a heteroalkyl group, wherein each alkyl group and heteroalkyl group is optionally substituted by one or more R 2 replace; L 2 is the key; M is absent, alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally replaced by one or more R 3 replace; P is optionally replaced by one or more R 4 substituted heteroaryl; Z is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally substituted by one or more R 5 replace; Each R A , R B , R C , R D , R E , R F and R G are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, azido, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally substituted by one or more R 6 replace; or R C and R D Together with the nitrogen atom to which they are attached, they form a 6 substituted rings (e.g., 5-7 membered rings); Each R 1 , R 2 , R 3 , R 4 , R 5 and R 6 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, –OR A1 , –C(O)OR A1 , –C(O)R B1 、–OC(O)R B1 , –N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 , –C(O)N(R C1 ), SR E1 、S(O) x R E1 、–OS(O) x R E1 , –N(R C1 )S(O) x R E1 , –S(O) x N(R C1 )(R D1 ),–P(R F1 ) y , cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally substituted by one or more R 7 replace; Each R A1 , R B1 , R C1 , R D1 , R E1 and R F1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by one or more R 7 replace; Each R 7 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxy, cycloalkyl or heterocyclyl; x is 1 or 2; and y is 2, 3, or 4; and (ii) an outer compartment comprising a second polysaccharide polymer comprising photoactive cross-linking moieties.

32. The hydrogel capsule of any one of claims 26-31, wherein the polysaccharide polymer (eg, the first polysaccharide polymer and / or the second polysaccharide polymer) is selected from the group consisting of alginate, hyaluronate, and chitosan.

33. The hydrogel capsule of any one of claims 26-32, wherein the polysaccharide polymer (eg, the first polysaccharide polymer and / or the second polysaccharide polymer) is alginate.

34. The hydrogel capsule of any one of claims 31-33, wherein the first polysaccharide polymer is alginate.

35. The hydrogel capsule of any one of claims 31-34, wherein the second polysaccharide polymer is alginate.

36. The hydrogel capsule of any one of claims 33-35, wherein the alginate is high guluronic acid (G) alginate or high mannuronic acid (M) alginate.

37. The hydrogel capsule of any one of claims 31-36, wherein the photoactive cross-linking moiety has formula (IV): (IV), or a pharmaceutically acceptable salt or tautomer thereof, wherein: X 1 Does not exist, O, NR 33 or C(R 34a )(R 34b ); R 30a , R 30b , R 31 , R 32 , R 33 , R 34a and R 34b Each of the above is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 , –C(O)OR A1 , –C(O)R B1 、–OC(O)R B1 , –N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 , –C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; Each R A1 , R B1 , R C1 , R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 replace; and Each R 7 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxy, cycloalkyl or heterocyclyl.

38. The hydrogel capsule of claim 37, wherein X 1 YesNR 33 (e.g., NH).

39. The hydrogel capsule of any one of claims 37-38, wherein R 30a , R 30b , R 31 and R 32 Each of is hydrogen.

40. The hydrogel capsule of any one of claims 31-39, wherein the photoactive cross-linking moiety has formula (IV-a): (IV-a), or a pharmaceutically acceptable salt or tautomer thereof, wherein: R 30a , R 30b , R 31 , R 32 and R 35 Each of the above is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 , –C(O)OR A1 , –C(O)R B1 、–OC(O)R B1 , –N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 , –C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; Each R A1 , R B1 , R C1 , R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 replace; and Each R 7 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxy, cycloalkyl or heterocyclyl.

41. The hydrogel capsule of claim 40, wherein R 30a , R 30b , R 31 , R 32 and R 35 Each of is hydrogen.

42. The hydrogel capsule of any one of claims 37-39, wherein the photoactive cross-linking moiety has formula (IV-c): (IV-c), or a pharmaceutically acceptable salt or tautomer thereof, wherein: R 30a , R 30b and R 31 Each of the above is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 , –C(O)OR A1 , –C(O)R B1 、–OC(O)R B1 , –N(R C1 )(R D1 ),–N(R C1 )C(O)R B1 , –C(O)N(R C1 ), SR E1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; R 32 is hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, –C(O)OR A1 , –C(O)R B1 , cycloalkyl, heterocyclyl, aryl or heteroaryl; Each R A1 , R B1 , R C1 , R D1 and R E1 are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted by 1-6 R 7 replace; and Each R 7 are independently alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxy, cycloalkyl or heterocyclyl.

43. The hydrogel capsule of claim 42, wherein R 30a , R 30b , R 31 and R 32 Each of is hydrogen.

44. The hydrogel capsule of any one of claims 31-43, wherein the photoactive cross-linking moiety has a structure selected from Table 4 or a pharmaceutically acceptable salt thereof.

45. The hydrogel capsule of any one of claims 31-44, wherein the photoactive cross-linking moiety is selected from acrylates, methacrylates, acrylamides and methacrylamides, or their corresponding acid chlorides and anhydrides.

46. ​​The hydrogel capsule of any one of claims 31-45, wherein the photoactive cross-linking moiety is selected from Compound 205 or Compound 217, or a pharmaceutically acceptable salt thereof.

47. The hydrogel capsule of any one of claims 31-46, wherein the compound of formula (I) has a structure selected from Table 3, or a pharmaceutically acceptable salt thereof.

48. The hydrogel capsule of any one of claims 31-46, wherein the compound of formula (I) is selected from Compound 100, Compound 101, Compound 110, Compound 112, Compound 113, Compound 114, Compound 122 and Compound 123, or a pharmaceutically acceptable salt thereof.

49. The hydrogel capsule of any one of claims 31-48, wherein the compound of formula (I) is Compound 101 or a pharmaceutically acceptable salt thereof.

50. The hydrogel capsule of any one of claims 31-49, wherein the hydrogel capsule has a diameter between 0.1 mm and 5 mm.

51. The hydrogel capsule of any one of claims 31-50, wherein the hydrogel capsule has a diameter between 1 mm and 5 mm.

52. The hydrogel capsule of any one of claims 31-51, wherein the hydrogel capsule has a diameter between 1 mm and 2.5 mm.

53. The hydrogel capsule of any one of claims 31-52, wherein the hydrogel capsule encapsulates cells.

54. The hydrogel capsule of claim 53, wherein the cells produce a therapeutic agent.

55. The hydrogel capsule of claim 54, wherein the therapeutic agent is a protein, such as a hormone, a coagulation factor, an antibody, or an enzyme.

56. The hydrogel capsule of any one of claims 31-55, wherein the hydrogel capsule is formulated for implantation into a subject (e.g., implantation into the intraperitoneal (IP) space, the abdominal cavity, the omentum, the lesser omental bursa, subcutaneous fat).

57. The hydrogel capsule of any one of claims 31-56, wherein the hydrogel capsule is formulated for implantation into the IP space of a subject.

58. A composition comprising the hydrogel capsule of any one of claims 31-57.

59. A method of producing a hydrogel capsule comprising the polysaccharide polymer of any one of claims 1-24.

60. A method of improving the stability of a hydrogel capsule comprising a polysaccharide polymer, wherein the method comprises providing means for ionically cross-linking the polysaccharide polymer and for covalently cross-linking the polysaccharide polymer.

61. The method of claim 60, wherein the means for ionically crosslinking the polysaccharide polymer comprises using divalent cations (e.g., Ba 2+、 Ca 2+ , Sr 2+ ).

62. The method of any one of claims 60-61, wherein the means for covalently cross-linking the polysaccharide polymer comprises the use of a photoactive cross-linking moiety (e.g., a vinyl cross-linking agent such as methacrylate, methacrylamide, or a pharmaceutically acceptable salt thereof).

63. A hydrogel capsule comprising: (i) an inner compartment comprising at least one cell and a polysaccharide polymer comprising a cell-binding peptide and optionally a photoactive cross-linking agent; and (ii) an outer compartment comprising a photoactive cross-linker as described herein and a compound of formula (I).

64. A hydrogel capsule comprising inner and outer compartments, said compartments comprising a polysaccharide polymer comprising a photoactive crosslinker and a compound of formula (I), wherein said inner compartment comprises cells.

65. A hydrogel capsule comprising: (i) an inner compartment comprising cells and a first polysaccharide polymer, optionally wherein the first polysaccharide polymer comprises a cell-binding peptide; and (ii) an outer compartment comprising a second polymer polysaccharide polymer comprising a photoactive cross-linking agent and a compound of formula (I).

66. A hydrogel capsule comprising: (i) an inner compartment comprising cells and a first polysaccharide polymer, wherein the first polysaccharide polymer is covalently modified with one or both of a photoactive cross-linking agent and a compound of formula (I); and (ii) an outer compartment comprising a second polymer polysaccharide polymer, wherein the second polysaccharide polymer is covalently modified with both a photoactive cross-linking agent and a compound of formula (I).

67. The hydrogel capsule of claim 66, wherein the first polysaccharide polymer and the second polysaccharide polymer are the same.

68. The hydrogel capsule of claim 66, wherein the first polysaccharide polymer and the second polysaccharide polymer are different.

69. The hydrogel capsule of any one of claims 66-68, wherein one or both of the inner compartments further comprises an unmodified polysaccharide polymer.

70. The hydrogel capsule of any one of claims 66-69, wherein the inner compartment further comprises a third polysaccharide polymer covalently modified with a cell-binding peptide.

71. The hydrogel capsule of claim 70, wherein the polymer in the third polysaccharide polymer comprises alginate.

72. A composition for treating a subject suffering from a disease, disorder or condition comprising the hydrogel capsule of any one of claims 26-57 and 63-71.

73. A composition for use as claimed in claim 72, wherein the disease, disorder or condition is diabetes (e.g., type 1 diabetes).

74. A composition for use as described in any one of claims 72-73, wherein the subject is a human.

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