Amino acid and amino alcohol based polyamine compounds and medical hydrogels formed therefrom

Abstract

In some aspects, the present disclosure relates to amino acid-based polyamine compounds formed by the ring-opening reaction of two or more amino acid N-carboxyl anhydrides molecules with a polyol having two or more hydroxyl groups under acid catalysis. In some aspects, the present disclosure relates to amino acid-based polyamine compounds formed by the ester coupling reaction of two or more amine-protected amino acid molecules with a polyol having two or more hydroxyl groups in the presence of an ester coupling agent. In some aspects, the present disclosure relates to amino alcohol-based polyamine compounds formed by the ester coupling reaction of two or more amine-protected amino alcohols molecules having one or more amino groups and a single hydroxyl group with a polycarboxylic acid compound having two or more carboxyl groups in the presence of an ester coupling agent.

Description

Cross-references to related applications

[0001] This application claims the benefit of U.S. Provisional Patent Application Serial No. 63 / 595,099, filed November 1, 2023, and U.S. Provisional Patent Application Serial No. 63 / 624,606, filed January 24, 2024, the disclosures of which are incorporated herein by reference. Technical Field

[0002] This disclosure relates to the use of polyamine compounds based on amino acids and amino alcohols as crosslinking agents in the formation of hydrogels, and to hydrogels formed from such compounds. The crosslinking agents and hydrogels can be used, for example, in various medical applications. Background Technology

[0003] SpaceOAR® is based on a multi-arm polyethylene glycol (PEG) polymer functionalized with succinimide glutarate as an active end group. Above a specific pH, this polymer rapidly crosslinks in vivo with lysine trimers (Lys-Lys-Lys) to form a hydrogel. This product has become a highly successful clinical biomaterial in prostate cancer treatment. A further improvement based on this structure involves functionalizing some of the succinimide glutarate end groups with 2,3,5-triiodobenzamide groups, thereby providing radiopermeability. This hydrogel is marketed as SpaceOAR® Vue, a radiopaque version of SpaceOAR® for prostate medical applications.

[0004] Although the current synthetic route for forming trilysine crosslinking agents is sufficient to produce monodisperse, well-defined small molecule oligomers, the method is verbose because it involves five steps and multiple protecting / deprotecting reactions.

[0005] For these and other reasons, alternative crosslinking agents are needed to form hydrogels and to form crosslinked hydrogels that provide enhanced radiation impermeability. Summary of the Invention

[0006] In some aspects, this disclosure provides amino acid-based polyamine compounds formed by (a) an amino acid N-carboxyanhydride molecule with (b) a polyol having two or more hydroxyl groups under acid-catalyzed ring-opening reaction.

[0007] In some embodiments, the amino acid N-carboxyl ring anhydride molecule is selected from glycine N-carboxyl ring anhydride, lysine N-carboxyl ring anhydride, ornithine N-carboxyl ring anhydride, and cysteine ​​N-carboxyl ring anhydride.

[0008] In some respects, this disclosure provides amino acid-based polyamine compounds formed by (a) ester coupling of an amino acid molecule protected by two or more amines with (b) a polyol having two or more hydroxyl groups in the presence of an ester coupling agent.

[0009] In some embodiments, the amine-protected amino acid is selected from amine-protected glycine, amine-protected lysine, amine-protected ornithine, and amine-protected cysteine.

[0010] In some aspects, this disclosure provides amino acid-based polyamine compounds comprising (a) residues of a polyol having two or more hydroxyl groups, and (b) two or more monoamine-terminated amino acid residues, each of which is linked by an ester bond at one of the two or more hydroxyl group residue sites.

[0011] In some embodiments, the amine-terminated amino acid residue is selected from amine-terminated glycine residues, amine-terminated lysine residues, amine-terminated ornithine residues, and amine-terminated cysteine ​​residues.

[0012] In some embodiments that can be used in any of the above aspects and implementations, the polyol has 3 to 20 hydroxyl groups.

[0013] In some aspects, this disclosure relates to amino alcohol-based polyamine compounds formed by ester coupling of (b) a polycarboxylic acid compound having two or more amino groups and a single hydroxyl group with an amine-protected amino alcohol molecule, preferably in the presence of an ester coupling agent.

[0014] In some embodiments, the amine-protected amino alcohol is an amine-protected C2-C alcohol having one, two, three, or four amino groups and a single hydroxyl group. 10 -Amino alcohol.

[0015] In some aspects, this disclosure relates to amino alcohol-based polyamine compounds comprising (a) residues of a polycarboxylic acid compound having two or more carboxyl groups, and (b) two or more monoamine-terminated amino alcohol residues, each of which has one or more amino groups and is connected by an ester bond at one of the two or more carboxyl groups.

[0016] In some embodiments, the amine-terminated amino alcohol residue is an amine-terminated C2-C [substance name missing]. 10 -Amino alcohol residues.

[0017] In some embodiments that can be used in the above aspects and implementations, the polycarboxylic acid has 3 to 20 carboxyl groups.

[0018] In other respects, this disclosure relates to systems for forming hydrogels comprising any of the above aspects and embodiments of an amino acid or amino alcohol-based polyamine compound and a polymer that forms a crosslink with the amino acid or amino alcohol-based polyamine compound.

[0019] In some embodiments, the polymer that crosslinks with the amino acid- or amino alcohol-based polyamine compound is a reactive multi-arm polymer comprising a plurality of hydrophilic polymer arms, each hydrophilic polymer arm comprising a hydrophilic polymer segment and a reactive end group, the reactive end group being covalently crosslinked with a primary amine group of the amino acid- or amino alcohol-based polyamine compound. In some of these embodiments, the hydrophilic polymer segment is selected from polyalkylene oxide segments, polyester segments, polyoxazoline segments, polydioxane-1,0-dioxane segments, and polypeptide segments. In some of these embodiments, the reactive group is an electrophilic group selected from imidazole esters, imidazole carboxylic esters, benzotriazole esters, or imide esters. In some of these embodiments, the reactive group is a cyclic imide ester group.

[0020] In some of these embodiments, the polymer that forms a crosslink with the polyamine compound based on amino acids or amino alcohols is an anionic polymer, comprising multiple anionic groups crosslinked with multiple primary amino ions of the polyamine compound based on amino acids or amino alcohols. In some of these embodiments, the anionic polymer comprises multiple carboxyl groups, sulfonate groups, sulfate groups, phosphate groups, or phosphonate groups.

[0021] In some aspects, this disclosure relates to medical hydrogels formed by crosslinking said amino acid or amino alcohol-based polyamine compounds with a polymer, said polymer being crosslinked with the amino acid or amino alcohol-based polyamine compounds of any of the above aspects and embodiments.

[0022] In some aspects, this disclosure relates to a treatment method comprising administering to a subject a mixture comprising an amino acid or amino alcohol-based polyamine compound and a polymer, wherein the polymer is cross-linked with the amino acid or amino alcohol-based polyamine compound under conditions that cause the amino acid or amino alcohol-based polyamine compound and the polymer to cross-link after administration.

[0023] In some aspects, this disclosure relates to a method for forming amino acid-based polyamine compounds, comprising a ring-opening reaction of an anhydride molecule containing two or more amino acid N-carboxyl rings with a polyol having two or more hydroxyl groups under acid-catalyzed conditions.

[0024] In some embodiments, the amino acid N-carboxyl ring anhydride molecule is a protected amino acid N-carboxyl ring anhydride molecule, and the method further includes a deprotection step after the ring-opening reaction.

[0025] In some embodiments that can be used in the above aspects and implementations, the reaction mixture contains an acid catalyst selected from HCl, phosphoric acid, p-toluenesulfonic acid, methanesulfonic acid and sulfuric acid.

[0026] In some embodiments that can be used in the above aspects and implementations, the ring-opening reaction is carried out with an excess of amino acid N-carboxyl ring anhydride molecules relative to the number of moles of hydroxyl groups in the reaction mixture.

[0027] In some embodiments that can be used in the above aspects and implementations, the ring-opening reaction is carried out with an excess of hydroxyl groups relative to the number of molar excess of anhydride molecules within the N-carboxyl ring of the amino acid in the reaction mixture.

[0028] In some aspects, this disclosure relates to a method for forming amino acid-based polyamine compounds, comprising ester coupling a two- or more amine-protected amino acid molecule with a polyol having two or more hydroxyl groups in the presence of an ester coupling agent; and a deprotection step following the ester coupling reaction.

[0029] In some implementations, the ester coupling agent is a carbodiimide coupling agent compound.

[0030] In some embodiments that can be used in the above aspects and implementations, the ester coupling reaction is carried out with an amino acid molecule protected by an amine in excess of a number of moles relative to the hydroxyl groups in the reaction mixture.

[0031] In some embodiments that can be used in the above aspects and implementations, the ester coupling reaction is carried out with an excess of hydroxyl groups relative to the number of molar excess amino acid molecules protected by the amine in the reaction mixture.

[0032] In some aspects, this disclosure relates to a method for forming an amino alcohol-based polyamine compound, comprising ester coupling a two- or more amine-protected amino alcohol molecule with a polycarboxylic acid molecule having two or more carboxyl groups in the presence of an ester coupling agent, and a deprotection step following the ester coupling reaction.

[0033] In some implementations, the ester coupling agent is a carbodiimide coupling agent.

[0034] In some embodiments that can be used in the above aspects and implementations, the ester coupling reaction is carried out with an amino alcohol molecule protected by an amine in excess relative to the number of moles of carboxyl groups in the reaction mixture.

[0035] In some embodiments that can be used in the above aspects and implementations, the ester coupling reaction is carried out with an excess of carboxyl groups relative to the number of molar excess amino alcohol molecules protected by the amine in the reaction mixture.

[0036] The above and other aspects, implementations, features and benefits of this disclosure will become apparent from the following detailed description. Attached Figure Description

[0037] Figure 1 The illustration schematically depicts a process for forming an amino acid-based polyamine compound from a polyol and an amino acid N-carboxyl ring anhydride, according to one embodiment of the present disclosure.

[0038] Figure 2 The process of forming an amino acid-based polyamine compound from glycerol and glycine N-carboxyl ring anhydride is illustrated schematically according to one embodiment of the present disclosure.

[0039] Figure 3 The process of forming an amino acid-based polyamine compound from inositol and glycine N-carboxyl ring anhydride is illustrated schematically according to one embodiment of the present disclosure.

[0040] Figure 4 The process of forming an amino acid-based polyamine compound from glycine protected by glycerol and amine is illustrated schematically according to one embodiment of the present disclosure.

[0041] Figure 5 The process of forming an amino acid-based polyamine compound from glycine protected by iodixanol and amine is illustrated schematically according to one embodiment of the present disclosure.

[0042] Figure 6 The process of forming an amino alcohol-based polyamine compound from N,N,N',N'-tetra(carboxymethyl)-ethane-1,2-diamine and amine-protected ethanolamine is illustrated schematically according to one embodiment of the present disclosure.

[0043] Figure 7 The process of forming an amino alcohol-based polyamine compound from ethanolamine protected by 2,4,6-triiodobenzene-1,3,5-tricarboxylic acid and an amine is illustrated schematically according to one embodiment of the present disclosure.

[0044] Figure 8 The process of forming an amino alcohol-based polyamine compound from 2-[bis(2-aminoethyl)amino]ethanol protected by 2,4,6-triiodobenzene-1,3,5-tricarboxylic acid and an amine is illustrated schematically according to a further embodiment of the present disclosure.

[0045] Figure 9 A delivery device according to an embodiment of the present disclosure is illustrated schematically. Detailed Implementation

[0046] In some aspects, this disclosure provides amino acid-based polyamine compounds formed by: (a) a ring-opening reaction of an amino acid N-carboxyl ring anhydride (NCA) with a polyol having two or more hydroxyl groups, or (b) an ester coupling reaction of an amine-protected amino acid with a polyol having two or more hydroxyl groups.

[0047] The amino acid-based polyamine compounds according to this disclosure comprise polyol residues and at least two amino acid residues, each of which is covalently linked to a polyol residue via an ester group.

[0048] In embodiments where an amino acid-based polyamine compound is formed by a ring-opening reaction of an NCA with a polyol having two or more hydroxyl groups, the amino acid residues comprise a primary amine group generated by the ring-opening process and a side group specific to each NCA.

[0049] In embodiments where amino acid-based polyamine compounds are formed by ester coupling of an amine-protected amino acid with a polyol having two or more hydroxyl groups, the amino acid residues also contain a primary amino group (after deprotection of the α-amino group) and side groups specific to each amino acid.

[0050] Regarding the ring-opening route, a single amino acid NCA reacts with a single hydroxyl group of a polyol under acid-catalyzed conditions without initiating (or desiring) further NCA polymerization. Therefore, at most one amino acid residue is attached to each hydroxyl site of the polyol. In this respect, by reacting with a molar excess of amino acid NCA relative to the total number of hydroxyl groups provided by the polyol in the reaction mixture, each hydroxyl group of the polyol reacts with a single amino acid NCA, and no hydroxyl groups remain in the resulting amino acid-based polyamine compound. On the other hand, by reacting with a molar excess of hydroxyl groups relative to the amino acid NCA, some hydroxyl groups of the polyol will remain unreacted in the resulting amino acid-based polyamine compound.

[0051] Acid catalysts that can be used in conjunction with the ring-opening reaction of this disclosure include HCl, phosphoric acid, p-toluenesulfonic acid, methanesulfonic acid, and sulfuric acid.

[0052] The amino acid N-carboxyl ring anhydride (NCA) used in this disclosure includes the formula... Among these, R is a side group unique to each amino acid's NCA. Amino acid NCAs include canonical NCAs and non-canonical NCAs. Specific examples of the R group are as follows: alanine (R= -CH3), cysteine ​​(R= -CH2SH), aspartic acid (R= -CH2COOH), glutamic acid (R= -CH2CH2COOH), phenylalanine (R= -CH2C6H5), glycine (R= -H), histidine (R=... Isoleucine (R= -CH(CH3)CH2CH3), Lysine (R= -(CH2)4NH2), Ornithine (R= -(CH2)3NH2), Leucine (R= -CH2CH(CH3)2), Methionine (R= -CH2CH2SCH3), Asparagine (R= -CH2CONH2), Glutamine (R= -CH2CH2CONH2), Arginine (R=-(CH2)3NH-C(NH)NH2), Serine (R= -CH2OH), Threonine (R= -CH(OH)CH3), Selenocysteine ​​(R= -CH2SeH), Valine (R= -CH(CH3)2), Tryptophan (R= ) and tyrosine (R=-CH2-C6H4OH), etc.

[0053] Regardless of which specific amino acid NCA is chosen, any amino acid residue attached to the polyol will have a free amino group at the end of the process.

[0054] Depending on the nature of the R group of the amino acid NCA, protecting groups may be required to prevent unwanted side reactions during ring-opening. For example, it may be necessary to protect amines, carboxylic acids, alcohols, or thiols during ring-opening. Examples of protecting groups used in conjunction with this disclosure include tert-butoxycarbonyl (Boc) groups, carboxybenzyl (Cbz) or (Z) groups, trifluoroacetyl (TFA) groups, 6-nitroveratryloxycarbonyl (Nvoc) groups, 9-fluorenylmethoxycarbonyl (Fmoc) groups, allyloxycarbonyl (Alloc) groups, triphenylmethyl (Trt) groups, tert-butyl (t-Bu) groups, benzyl ester (β-benzyl) groups, O-benzyl, S-ethylsulfonyl, 2,4-dimethoxybenzyl (Dmb), tert-butyldimethylsilyl (TBDMS), allyl, o-nitrobenzyl (PNB), p-methylbenzyl (Meb), and acetaminomethyl (Acm) groups, etc.

[0055] In some embodiments, amino acid NCAs containing protected amine side groups are used. For example, amino acid NCAs containing a protected primary amine side group, such as an aminoalkyl group (e.g., C1-C6-aminoalkyl, including aminomethyl, 2-aminoethyl, 3-aminopropyl, 4-aminobutyl, 5-aminopentyl, and 6-aminohexyl, and isomers thereof) and a protected imidazole side group can be used. Specific examples include amine-protected lysine NCA monomers (where the protecting group is 4-aminobutyl), amine-protected ornithine NCA monomers (where the protecting group is 3-aminopropyl), and amine-protected histidine NCA monomers (where the protecting group is imidazole), etc. More specific examples of such amine-protected amino acid NCAs include lysine (Boc)-NCA. Ornithine (Boc-NCA) Lysine (Z)-NCA, also known as lysine (Cbz)-NCA, Ornithine (Z)-NCA, also known as ornithine (Cbz)-NCA, And lysine (TFA)-NCA, N(Im)-(2,4-dinitrophenyl)-L-histidine NCA. Other examples of amino acid NCAs include protected amino acid NCAs containing a protected carboxyl side group (e.g., C1-C6-carboxylalkyl, including carboxymethyl, 2-carboxyethyl, 3-carboxypropyl, 4-carboxybutyl, 5-carboxypentyl, and 6-carboxyhexyl, and their isomers), such as β-benzyl-L-aspartic acid NCA. , and γ-benzyl-L-glutamic acid NCA, Amino acid NCAs containing protected hydroxyl side groups (e.g., C1-C6-hydroxyalkyl, including hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl, 4-hydroxybutyl, 5-hydroxypentyl, and 6-hydroxyhexyl, as well as their isomers and phenolic side groups), such as O-benzyl-L-serine NCA, , and O-benzyl-L-tyrosine NCA, And protected amino acid NCAs containing protected thiol side groups (e.g., C1-C6-mercaptoalkyl, including mercaptomethyl, 2-mercaptoethyl, 3-mercaptopropyl, 4-mercaptobutyl, 5-mercaptopentyl, and 6-mercaptohexyl, and their isomers), such as S-tert-butylmercaptocysteine ​​NCA. ,etc.

[0056] After the acid-catalyzed ring-opening process, the resulting protected amino acid residues can be protected using known techniques.

[0057] Now we turn to the ester coupling route, where each hydroxyl group of the polyol reacts with a single amino acid by reacting it with an amine-protected amino acid in a molar excess relative to the total number of hydroxyl groups provided by the polyol in the reaction mixture, resulting in no hydroxyl residues in the resulting amino acid-based polyamine compound. Alternatively, by reacting with a molar excess of hydroxyl groups relative to the amine-protected amino acid, some hydroxyl groups of the polyol will remain unreacted in the resulting amino acid-based polyamine compound.

[0058] Ester coupling between a carboxylic acid and a hydroxyl group can be carried out in the presence of a suitable coupling agent, such as a carbodiimide coupling agent, like N,N'-dicyclohexylcarbodiimide (DCC), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), or N,N'-diisopropylcarbodiimide (DIC). The formed ester bond is hydrolyzable.

[0059] The amino acids used in this article for amine protection include both normatively amine-protected and non-normatively amine-protected amino acids.

[0060] The protecting groups of the α-amino group of amine-protected amino acids include tert-butyloxycarbonyl (Boc) groups, 9-fluorenylmethoxycarbonyl (Fmoc) groups, tert-butyl (t-Bu) groups, and benzyl (Bn) groups. When using a Boc protecting group, the amine-protected amino acids used in this disclosure include those of the formula... Those, where R is a side group specific to each protected amino acid. When using the Fmoc protecting group, the amino acids used for amine protection in this disclosure include those of the formula... Those, where R is a side group specific to each protected amino acid. Examples of the R group, as mentioned above, are: glycine (R= -H), alanine (R= -CH3), cysteine ​​(R= -CH2SH), aspartic acid (R= -CH2COOH), glutamic acid (R= -CH2CH2COOH), phenylalanine (R= -CH2C6H5), histidine (R= Isoleucine (R=-CH(CH3)CH2CH3), Lysine (R=-(CH2)4NH2), Ornithine (R=-(CH2)3NH2), Leucine (R=-CH2CH(CH3)2), Methionine (R=-CH2CH2SCH3), Asparagine (R=-CH2CONH2), Glutamine (R=-CH2CH2CONH2), Arginine (R=-(CH2)3NH-C(NH)NH2), Serine (R=-CH2OH), Threonine (R=-CH(OH)CH3), Selenocysteine ​​(R=-CH2SeH), Valine (R=-CH(CH3)2), Tryptophan (R=-CH(CH3)2), ) and tyrosine (R=-CH2-C6H4OH), etc.

[0061] Regardless of the specific amino acid chosen, any amino acid residue attached to a polyol will have at least one free amino group at the end of the deprotection process.

[0062] Depending on the nature of the R group of the amino acid, additional protecting groups may be required to prevent unwanted R reactions during ester coupling. For example, during ester coupling, it may be necessary to protect non-α-amino R groups (such as aminoalkyl groups (e.g., C1-C6-aminoalkyl groups, including aminomethyl, 2-aminoethyl, 3-aminopropyl, 4-aminobutyl, 5-aminopentyl, and 6-aminohexyl, and their isomers) (e.g., for lysine, ornithine, etc.), carboxylic acid R groups (such as carboxylalkyl groups (e.g., C1-C6-carboxylalkyl groups, including carboxymethyl, 2-carboxyethyl, 3-carboxypropyl, 4-carboxybutyl, 5-carboxypentyl, and 6-carboxyhexyl, and their isomers) (e.g., for aspartic acid, glutamic acid, etc.), hydroxy (alcohol) R groups (e.g., for serine, threonine, hydroxyproline, tyrosine), thiol R groups (e.g., for cysteine), guanidino (e.g., for arginine), or imidazole (e.g., for histidine).

[0063] Examples of other protecting groups used in conjunction with this disclosure include protecting groups for amine R groups (such as tert-butoxycarbonyl (Boc) groups, carboxybenzyl (Cbz) groups, allyloxycarbonyl (Alloc) groups, or 6-nitroveratroloxycarbonyl (Nvoc) groups), protecting groups for carboxyl R groups (such as triphenylmethyl (Trt), tert-butyl (t-Bu), benzyl (Bn), 9-fluorenylmethyl (Fm) or 2,4-dimethoxybenzyl (Dmb) groups), protecting groups for hydroxyl R groups (such as tert-butyldimethylsilyl (TBDMS), allyl or o-nitrobenzyl (ONB) groups), and protecting groups for guanidine R groups (such as 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl residues (Pbf)). Protected groups include those for imidazole R groups (such as toluenesulfonyl (Tos) or benzyloxymethyl (Bom) groups), and those for thiol R groups (such as p-methylbenzyl (Meb), acetaminomethyl (Acm), and triphenylmethyl (Trt) groups). In specific cases where the amino R group is protected, commercially available compounds include protected lysine, such as Boc-Lys(Boc)-OH. (CAS#15098-69-8) and protected ornithines, such as Boc-Orn(Boc)-OH, (CAS#57133-29-6).

[0064] After the ester coupling process, the resulting protected amino acid residues can be protected using known techniques.

[0065] Polyols that can be used in conjunction with the ring-opening and ester coupling reactions described herein include those having two or more hydroxyl groups, for example, containing 2 to 100 hydroxyl groups (e.g., having 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, or 100 hydroxyl groups). General categories of molecules include sugars (monosaccharides, disaccharides, trisaccharides, etc.), sugar alcohols, calixarenes, polyhedral oligosemisilsesquioxanes (POSS), cyclodextrins, polyhydroxylated polymers, catechins, flavanols, anthocyanins, stilbenes, and polyphenols. Generally, this disclosure relates to alcohols, including primary alcohols, secondary alcohols, tertiary alcohols, and phenols, that react with amino acid NCAs when initiated by an acid catalyst.

[0066] Exemplary polyols include straight-chain, branched, and cyclic sugars and sugar alcohols, such as glycerol, mannitol, sorbitol, inositol, xylitol, styritol, arabinol, erythritol, pentaerythritol, dipentaerythritol, tripentaerythritol, arbutinol, hexaglycerol, eugenol, fucose, ribose, arabinose, xylose, lysose, rhamnose, galactose, glucose, fructose, sorbitol, mannose, pyranose, azoose, tarose, tagatose, pyranoside, sucrose, lactose, and maltose; short polymers (including dimers, trimers, tetramers, pentamers, hexamers, heptomers, octamers, nonamermers, and decamers) of straight-chain, branched, and cyclic sugars and sugar alcohols (including the aforementioned sugars and sugar alcohols); starch, amylose, dextrin, cyclodextrin; and polyhydroxy crown ethers and polyhydroxy alkyl crown ethers. Exemplary polyols also include aromatic polyols, including 1,1,1-tris(4'-hydroxyphenyl)alkanes, such as 1,1,1-tris(4-hydroxyphenyl)ethane and 2,6-bis(hydroxyalkyl)cresol.

[0067] Exemplary polyols also include polyhydroxylated polymers. For example, in some embodiments, the core region comprises polyhydroxylated polymer residues, such as poly(vinyl alcohol) residues, poly(allyl alcohol), poly(hydroxyethyl acrylate) residues, or poly(hydroxyethyl methacrylate) residues. The length of such polyhydroxylated polymer residues can range, for example, from 2 to 100 monomer units.

[0068] Specific examples of polyols include glycerol, (n=3, where n is the number of -OH functional groups in the molecule), 1,2,4-butanetriol, (n=3), resveratrol (n=3), 3-(2-hydroxyethyl)pentane-1,5-diol, (n=3), Triethanolamine Inositol (n=6), sorbitol, (n = 6), Tripentaerythritol, (n=8), epicatechin, (n=5), epicatechin gallate (n=7), maltohexaose, (n=20), and cyclodextrins with 4-10 glucose units (n=12-30), etc.

[0069] In some embodiments, iodinated polyols can be used to provide radiation impermeability to the resulting polyamine compound. In some of these embodiments, the iodinated polyol is a compound comprising two or more hydroxyl groups and one or more iodinated aromatic groups.

[0070] Examples of iodinated aromatic groups include iodine-substituted monocyclic aromatic groups and iodine-substituted polycyclic aromatic groups, such as iodine-substituted phenyl, iodine-substituted naphthyl, iodine-substituted anthraceneyl, iodine-substituted phenanthryl, and iodine-substituted tetraphenyl. The aromatic group can be substituted with one, two, three, four, five, six, or more iodine atoms. In various embodiments, the aromatic group is further substituted with two or more hydroxyl groups, which can directly replace the aromatic group or can be provided in the form of a hydroxyalkyl group (e.g., a C1-C4 hydroxyalkyl group containing one, two, three, or four carbon atoms and containing one, two, three, or four or more hydroxyl groups). The hydroxyalkyl group can be attached to the aromatic group directly or via any suitable linking moiety selected from, for example, amide, amino, ether, ester, or carbonate groups.

[0071] Specific examples of iodinated polyols used in this disclosure include known iodinated contrast agents whose biocompatibility has been demonstrated to be quite well tolerated. Exemplary iodinated polyols include 2,4,6-triiodophenyl-1,3,5-triol. (n=3), Iopromide, (n=4), iopamidol, (n=5), Iohexol, (n=6), Iodophorol, (n=6) and iodixanol, (n=9).

[0072] In other embodiments, radiation impermeability can be introduced into polyamine compounds by using iodinated amino acid NCA derivatives in ring-opening synthesis or by using amine-protected iodinated amino acids in ester coupling reactions. For example, in some embodiments, iodinated phenylalanine NCA (e.g., 4-iodo-L-phenylalanine NCA) and iodinated tyrosine NCA (e.g., 3-iodo-L-tyrosine NCA) can be used in ring-opening synthesis, and in some embodiments, amine-protected iodinated phenylalanine (e.g., protected 4-iodo-L-phenylalanine) or protected iodinated tyrosine (e.g., protected 3-iodo-L-tyrosine) can be used in ester coupling reactions.

[0073] It is noteworthy that using NCA and amine-protected amino acids containing protected amine side groups, such as those based on lysine or ornithine, effectively doubles the number of amine functional groups that can be attached to any given molecule. For example, a molecule with n = 3, as shown above, will be able to have six additional amine functional groups. This means that the maximum number m of amine functional groups that can be attached to a given polyol ranges from m = n (where amino acids without protected amine side groups are used) to m = 2n (where amino acids with protected amine side groups are used). As previously mentioned, for example, using fewer amino acids in a molar excess relative to the hydroxyl group can result in fewer than the maximum number of amine functional groups. This is reflected in the table below.

[0074] We will now discuss various specific open-ring implementation schemes and ester coupling implementation schemes.

[0075] Now for reference Figure 1 The general procedure is shown, in which a polyol (112) having n hydroxyl groups (where R is an organic group representing the remainder of the polyol) is combined with m amino acid NCA molecules (114) (where R... 1 The reaction (representing a canonical or non-canonical amine group protected by a protected group such as Cbz, Boc, TFA, Nvoc, or FMOC) is followed by deprotection of the protected group to form a polyamine compound with m additional amino acid residues and nm unreacted hydroxyl groups (116).

[0076] Now for reference Figure 2 The specific procedure is shown in which 1 equivalent of glycerol (212) having 3 hydroxyl groups reacts with 3 equivalents of glycine NCA to form a polyamine compound (216) having 3 additional glycine residues, which are linked to the glycerol residues via ester bonds.

[0077] Similarly, refer to Figure 3Another specific procedure is shown in which 1 equivalent of inositol (312) having 6 hydroxyl groups reacts with 6 equivalents of glycine NCA to form a polyamine compound (316) having 6 additional glycine residues, which are linked to the inositol residues via ester bonds.

[0078] As an example of an ester coupling strategy, see [reference]. Figure 4 In the presence of a carbodiimide coupling agent such as DCC, glycerol (412) reacts with excess Boc-glycine (414) to produce 2,3-bis[[2-(tert-butoxycarbonylamino)acetyl]oxy]propyl 2-(tert-butoxycarbonylamino)acetate (416). This intermediate is then treated with a dioxane solution of a deprotecting agent, such as HCl, to give 2,3-bis[(2-aminoacetyl)oxy]propyl 2-aminoacetate (418), a polyamine compound having three additional glycine residues linked to glycerol residues via ester bonds.

[0079] Figure 5 A similar reaction using an iodinated polyol is shown, wherein iodixanol (512) reacts with excess Boc-glycine (514) in the presence of a carbodiimide coupling agent such as DCC to form a Boc-protected iodinated intermediate (516), which is then treated with a dioxane solution of a deprotecting agent such as HCl to give the nonylamine-functionalized iodide final product (418), which is a polyamine compound having nine additional glycine residues that are linked to glycerol residues via ester bonds.

[0080] In various respects, this disclosure also provides amino alcohol-based polyamine compounds formed by ester coupling of (a) an amine-protected amino alcohol with a polycarboxylic acid compound having two or more carboxyl groups.

[0081] The amino alcohol-based polyamine compound according to this disclosure comprises a polycarboxylic acid residue and at least two amino alcohol residues, each of which is covalently linked to the polycarboxylic acid residue via an ester group.

[0082] Ester coupling is performed by using an amine-protected amino alcohol in a molar excess relative to the total number of carboxyl groups provided by the polycarboxylic acid in the reaction mixture. This allows each carboxyl group of the polycarboxylic acid to react with a single amine-protected amino alcohol, resulting in no carboxyl group residue in the resulting amino alcohol-based polyamine compound. Alternatively, by reacting with a molar excess of carboxyl groups relative to the amine-protected amino alcohol, some carboxyl groups of the polycarboxylic acid will remain unreacted in the resulting amino alcohol-based polyamine compound.

[0083] Ester coupling between a carboxylic acid and a hydroxyl group can be carried out in the presence of suitable coupling agents, such as carbodiimide coupling agents, including N,N'-dicyclohexylcarbodiimide (DCC), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), or N,N'-diisopropylcarbodiimide (DIC). The resulting ester bond is hydrolyzable.

[0084] The protecting groups of amine-protected amino alcohols include tert-butyloxycarbonyl (Boc) group, 9-fluorenylmethoxycarbonyl (Fmoc) group, carboxybenzyl (Cbz) group, tert-butyl (t-Bu) group, and benzyl (Bn) group.

[0085] After the ester coupling process, the amino alcohol residues protected by the resulting amine can be protected using known techniques.

[0086] The amino alcohols used in this disclosure include amino alcohols having one hydroxyl group and one, two, three, four, five, six, seven, eight or more amino groups (e.g., C1-amino alcohol, C2-amino alcohol, C3-amino alcohol, C4-amino alcohol, C5-amino alcohol, C6-amino alcohol, C7-amino alcohol, C8-amino alcohol, C9-amino alcohol, C...). 10 -Amino alcohol, C 11 -Amino alcohol, C 12 -Amino alcohol, C 13 -Amino alcohol, C 14 -Amino alcohol, C 15 -Amino alcohol, C 16 -Amino alcohol, C 17 -Amino alcohol, C 18 -Amino alcohol, C 19 -Amino alcohol, C 20 -Amino alcohols, etc.). The following lists various C2-C8-amino alcohols having one, two, or three amino groups.

[0087] Regardless of the specific amino alcohol chosen, at the end of the deprotection process, any amino alcohol residue attached to the polycarboxylic acid will have at least one free amino group.

[0088] The polycarboxylic acids used according to this disclosure include polycarboxylic acids having two or more carboxylic acid groups, for example, containing 2 to 100 carboxylic acid groups (e.g., having 2 to 3 to 4 to 5 to 6 to 7 to 8 to 9 to 10 to 12 to 15 to 20 to 25 to 30 to 40 to 50 to 60 to 70 to 80 to 90 to 100 carboxylic acid groups).

[0089] Exemplary polycarboxylic acids can be selected from, for example, non-iodinated polycarboxylic acids having two or more carboxylic acid groups, including dicarboxylic acids such as C1-C8 alkane dicarboxylic acids (including α, ω-C2-C8 alkane dicarboxylic acids, such as 1,2-ethanedicarboxylic acid, 1,3-propanedicarboxylic acid, 1,4-butanedicarboxylic acid, 1,5-pentanedicarboxylic acid, 1,6-hexanedicarboxylic acid, etc.), tricarboxylic acids (such as propane-1,2,3-tricarboxylic acid, benzene-1,3,5-tricarboxylic acid), tetracarboxylic acids, pentacarboxylic acids, hexacarboxylic acids, heptacarboxylic acids, octacarboxylic acids, etc. Non-iodinated polycarboxylic acids having two or more carboxylic acid groups can also be formed from any of the above-mentioned non-iodinated polyols having two or more hydroxyl groups by reacting the hydroxyl groups of the polyol with cyclic anhydrides to form carboxylic acid groups.

[0090] Further illustrative polycarboxylic acids include iodinated polycarboxylic acids. Iodinated polycarboxylic acids include iodinated aromatic polycarboxylic acids, examples of which are compounds comprising two or more carboxylic acid groups and one or more iodinated aromatic groups. Examples of iodinated aromatic groups include iodine-substituted monocyclic aromatic groups and iodine-substituted polycyclic aromatic groups, such as iodine-substituted phenyl, iodine-substituted naphthyl, iodine-substituted anthraceneyl, iodine-substituted phenanthryl, and iodine-substituted tetraphenyl, etc. The aromatic group may be substituted with one, two, three, four, five, six, or more iodine atoms. In various embodiments, the aromatic group is further substituted with two or more carboxylic acid groups, which may directly replace the aromatic group or may be provided in the form of a carboxyalkyl group (e.g., a C1-C4 carboxyalkyl group containing one, two, three, or four carbon atoms and containing one, two, three, or four or more carboxylic acid groups). The carboxyalkyl group may be attached to the aromatic group directly or via any suitable linking moiety selected from, for example, amide, amino, ether, ester, or carbonate groups.

[0091] Iodinated polycarboxylic acids having two or more carboxylic acid groups can be formed from any of the above-mentioned iodinated polyols having two or more hydroxyl groups by reacting the hydroxyl groups of the polyol with cyclic anhydrides to form carboxylic acid groups.

[0092] The following table shows various C2-C groups having 2 to 8 carboxyl groups. 50 - Polycarboxylic acids.

[0093] It is noteworthy that using amine-protected amino alcohols containing two protected amine groups (e.g., 1-amino-3-(2-aminoethoxy)-2-propanol, 2,2-bis(2-aminoethoxy)ethanol, 2-[bis(2-aminoethyl)amino]ethanol, 5-amino-2-(3-aminopropyl)-1-pentanol, etc.) effectively doubles the number of amine functional groups that can be attached to any given molecule. For example, as shown above, a molecule with q = 2 will be able to have four additional amine functional groups, a molecule with q = 3 will be able to have six additional amine functional groups, a molecule with q = 4 will be able to have eight additional amine functional groups, a molecule with q = 5 will be able to have ten additional amine functional groups, a molecule with q = 6 will be able to have twelve additional amine functional groups, and so on.

[0094] Similarly, using amine-protected amino alcohols containing three protected amino groups (e.g., 1-amino-3-[2-amino-1-(aminomethyl)ethoxy]-2-propanol, etc.) effectively triples the number of amine functional groups that can be attached to any given molecule. For example, a molecule with q = 2, as shown above, will be able to have six additional amine functional groups, a molecule with q = 3 will be able to have nine additional amine functional groups, a molecule with q = 4 will be able to have twelve additional amine functional groups, a molecule with q = 5 will be able to have fifteen additional amine functional groups, a molecule with q = 6 will be able to have eighteen additional amine functional groups, and so on.

[0095] We will now discuss various specific ester coupling implementation schemes.

[0096] The reaction between polycarboxylic acids and protected amino alcohols is as follows: Figure 6 As shown, it schematically illustrates the process in which N,N,N',N'-tetra(carboxymethyl)-ethane-1,2-diamine (612) reacts with excess N-Boc-ethanolamine (614) in the presence of a carbodiimide coupling agent such as DCC to form a Boc-protected intermediate (616), which is then treated with a dioxane solution of a deprotecting agent such as HCl to produce a tetraamine-functionalized final product (618), which is a polyamine compound having four additional ethanolamine residues that are linked to N,N,N',N'-tetra(carboxymethyl)-ethane-1,2-diamine residues via ester bonds.

[0097] The reaction between iodinated polycarboxylic acids and protected amino alcohols is as follows: Figure 7As shown, it schematically illustrates the process in which 2,4,6-triiodobenzene-1,3,5-tricarboxylic acid (712) reacts with excess N-Boc-ethanolamine (714) in the presence of a carbodiimide coupling agent such as DCC to form a Boc-protected iodinated intermediate (716), which is then treated with a dioxane solution of a deprotecting agent such as HCl to produce a final product (718) functionalized with triiodinated triamine, which is a polyamine compound having three additional ethanolamine residues that are linked to 2,4,6-triiodobenzene-1,3,5-tricarboxylic acid residues via ester bonds.

[0098] The reaction between iodinated polycarboxylic acids and protected amino alcohols having multiple amino groups is as follows: Figure 8 As shown, it schematically illustrates that 2,4,6-triiodophenyl-1,3,5-tricarboxylic acid (812) reacts with excess Boc-protected 2-[bis(2-aminoethyl)amino]ethanol (814) in the presence of a carbodiimide coupling agent such as DCC to form a Boc-protected iodinated intermediate (816), which is then treated with a dioxane solution of a deprotecting agent such as HCl to produce a final product (818) functionalized with hexylamine iodide, which is a polyamine compound having three additional 2-[bis(2-aminoethyl)amino]ethanol residues, each residue having two amino groups, which are linked to the 2,4,6-triiodophenyl-1,3,5-tricarboxylic acid residues via ester bonds.

[0099] The amino acid and amino alcohol-based polyamine compounds according to this disclosure can be used in a variety of medical applications. In some embodiments, the amino acid and amino alcohol-based polyamine compounds can be used as crosslinking agents, for example, as covalent crosslinking agents, ionic crosslinking agents, or hydrogen bond-based crosslinking agents.

[0100] In this regard, various aspects of this disclosure provide cross-linked hydrogels comprising (a) a polyamine compound based on an amino acid or an amino alcohol, such as one of those described above, and (b) a cross-linking reaction product of a polymer cross-linked with the polyamine compound based on an amino acid or an amino alcohol. Such cross-linked hydrogels can be formed in vivo (e.g., using the delivery device described below), or they can be formed in vitro and subsequently administered to a subject. Such cross-linked hydrogels can be used in a variety of biomedical applications, including medical devices, implants, and pharmaceutical compositions.

[0101] In embodiments of the amino acid and amino alcohol-based polyamine compounds of this disclosure that incorporate radiopermeability, the cross-linked hydrogels are visible under a fluorescence microscope. In various embodiments, such cross-linked hydrogels have radiopermeability of 100 Huntsfield units (HU) or higher, advantageously in the range of 100 HU to 250 HU to 500 HU to 750 HU to 1000 HU to 2500 HU or higher.

[0102] In some embodiments, the polymer that crosslinks with the amino acid- or amino alcohol-based polyamine compound is a reactive polymer that contains a plurality of reactive groups that covalently crosslink with the primary amine group of the amino acid- or amino alcohol-based polyamine compound.

[0103] Examples of reactive polymers comprising multiple reactive groups that covalently crosslink with the primary amine group of a polyamine compound based on an amino acid or amino alcohol include multi-arm polymers comprising multiple polymer arms (e.g., having two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, or more arms), wherein two or more polymer arms of the multi-arm polymer comprise one or more reactive end groups. In some embodiments, compositions containing reactive multi-arm polymers may be provided, wherein the percentage of polymer arms comprising one or more reactive end groups may correspond to between 50% and 100% of the total number of polymer arms in the composition (e.g., any range from 50% to 70% to 80% to 90% to 95% to 99% to 100% of the total number of polymer arms).

[0104] In various embodiments, the polymer arms of the multi-arm polymer are hydrophilic polymer arms, and each polymer arm contains a hydrophilic polymer segment.

[0105] The hydrophilic polymer segments used for the polymer arm can be selected from a variety of synthetic, natural, or synthetic-natural hybrid hydrophilic polymer segments. Examples of hydrophilic polymer segments include those formed from one or more hydrophilic monomers selected from: C1-C6 epoxides (e.g., ethylene oxide, propylene oxide, tetramethylene oxide, etc.), polar aprotic vinyl monomers (e.g., N-vinylpyrrolidone, acrylamide, N-methacrylamide, dimethacrylamide, N-vinylimidazolium, 4-vinylimidazolium, sodium 4-vinylbenzenesulfonate, etc.), p-dioxanone, ester monomers (e.g., glycolide, lactide, β-propiolactone, β-butyrolactone, γ-butyrolactone, γ-... Valprolactone, δ-valprolactone, ε-caprolactone, etc.), oxazoline monomers (e.g., oxazoline and 2-alkyl-2-oxazoline, such as 2-(C1-C6 alkyl)-2-oxazoline, including various isomers such as 2-methyl-2-oxazoline, 2-ethyl-2-oxazoline, 2-n-propyl-2-oxazoline, 2-isopropyl-2-oxazoline, 2-n-butyl-2-oxazoline, 2-isobutyl-2-oxazoline, 2-hexyl-2-oxazoline, etc.), 2-phenyl-2-oxazoline, N-isopropylacrylamide, amino acids and sugars.

[0106] The hydrophilic polymer segments can be selected from, for example, the following polymer segments: polyether segments (including poly(C1-C6-epoxy) segments, such as poly(ethylene oxide)(PEO) (also known as polyethylene glycol or PEG) segments, poly(propylene oxide) segments, poly(ethylene oxide-copolymer-propylene oxide) segments), polymer segments formed from one or more polar aprotic vinyl monomers (including poly(N-vinylpyrrolidone) segments, poly(acrylamide) segments, poly(N-methacrylamide) segments, poly(dimethacrylamide) segments, poly(N-vinylimidazolium) segments, poly(4-vinylimidazolium) segments, and poly(4-vinylbenzenesulfonate) segments), polydioxane-hexanone segments, polyester segments (including polyglycolic acid lactide segments, ... Polylactide segments, poly(lactide-co-glycolic acid) segments, poly(β-propiolactone) segments, poly(β-butyrolactone) segments, poly(γ-butyrolactone) segments, poly(γ-valerolactone) segments, poly(δ-valerolactone) segments and poly(ε-caprolactone) segments, polyoxazoline segments (including poly(2-C1-C6-alkyl-2-oxazoline segments), such as poly(2-methyl-2-oxazoline) segments, poly(2-ethyl-2-oxazoline) segments, poly(2-propyl-2-oxazoline) segments, poly(2-isopropyl-2-oxazoline) segments, and poly(2-n-butyl-2-oxazoline) segments, poly(2-phenyl-2-oxazoline) segments, poly(N-isopropylacrylamide) segments, polypeptide segments and polysaccharide segments. Polysaccharide segments include those containing one or more uronic acid substances, such as galacturonic acid, glucuronic acid and / or iduronic acid. Specific examples of polysaccharide segments include alginate, hyaluronic acid, pectin, agar, carrageenan, gellan gum, gum arabic, guar gum, xanthan gum and carboxymethyl cellulose.

[0107] The polymer segments used in the multi-arm polymers of this disclosure typically contain 10 to 1000 monomer units.

[0108] In some embodiments, the polymer arms of the reactive multi-arm polymer extend from the core region. In some of these embodiments, the core region includes residues of a polyol for forming the polymer arms. Exemplary polyols may be selected from, for example, the polyols described above for forming the amino acid-based polyamine compounds of this disclosure.

[0109] In some embodiments, the reactive end group is an electrophilic group. The electrophilic group can be selected, for example, from cyclic imide ester groups, such as succinimide ester groups. Maleimide ester group, glutarimide ester group, diglycolimide ester group, phthalimide ester group and bicyclic [2.2.1]hept-5-ene-2,3-dicarboxylic acid imide ester group Imidazole ester group, imidazole carboxylic ester group and benzotriazole ester group, etc.

[0110] Electrophilic groups can be attached to hydrophilic polymer segments through any suitable linker, which can be selected from, for example, linkers containing alkyl groups, linkers containing ether groups, linkers containing ester groups, linkers containing amide groups, linkers containing amine groups, linkers containing carbonate groups, or linkers containing a combination of two or more of the aforementioned groups.

[0111] In some embodiments, the linker portion comprises a hydrolyzable ester group. The hydrolyzable ester group may be selected from, for example, glutarate, succinate, carbonate, or adipate groups. In specific embodiments, the polymer arm may be capped with reactive, hydrolyzable, or other groups such as succinimidyl glutarate, succinimidyl succinate, succinimidyl carbonate, or succinimidyl adipate.

[0112] In other embodiments, the reactive multi-arm polymer does not contain hydrolyzable ester groups. In this regard, it is noted that the amino acid and amino alcohol-based polyamine compounds provided herein have hydrolyzable ester groups.

[0113] For example, further examples of reactive multi-arm polymers are described in U.S. Patent Applications Nos. 2011 / 0142936, 2021 / 0061950, 2021 / 0061954, and 2021 / 0061957.

[0114] In a specific example, the reactive multi-arm polymer described above, comprising a core region and a plurality of hydrophilic polyethylene oxide arms (specifically, eight arms) having reactive end groups (e.g., succinimidyl groups), can be covalently crosslinked with an amino acid- or amino alcohol-based polyamine compound as described above, the polyamine compound comprising primary amine groups that react with the reactive end groups of the reactive multi-arm polymer. A crosslinked product is formed by reacting the amino acid- or amino alcohol-based polyamine compound with the reactive multi-arm polymer under alkaline conditions.

[0115] In some embodiments, the polymer that crosslinks with the amino acid- or amino alcohol-based polyamine compound can be a poly(carboxylic acid) that covalently crosslinks with the amino acid- or amino alcohol-based polyamine compound, such as poly(acrylic acid), poly(methacrylic acid), alginate, hyaluronic acid, pectin, agar, carrageenan, gellan gum, gum arabic, guar gum, xanthan gum, or carboxymethyl cellulose. These and other poly(carboxylic acids) have carboxylic acid groups that can form amide bonds with the primary amine groups of the amino acid- or amino alcohol-based polyamine compound. This crosslinking can be formed, for example, in the presence of a carbodiimide coupling agent. In a specific example, hyaluronic acid can be covalently crosslinked with the amino acid- or amino alcohol-based polyamine compound described herein, the polyamine compound comprising a primary amine group that forms an amide bond with the carboxylic acid group of hyaluronic acid, thereby forming a crosslinked product.

[0116] In some embodiments, the polymer that crosslinks with the amino acid- or amino alcohol-based polyamine compound is an anionic polymer containing a plurality of anionic groups capable of crosslinking with the cationic primary amine ions of the amino acid- or amino alcohol-based polyamine compound. In some embodiments, the anionic polymer is water-soluble.

[0117] Examples of anionic polymers comprising multiple anionic groups capable of crosslinking with cationic primary amine groups of polyamine compounds based on amino acids or amino alcohols include any variety of synthetic, natural, or synthetic-natural hybrid anionic polymers containing one or more groups selected from carboxyl groups, sulfonate groups, sulfate groups, phosphate groups, or phosphonate groups, which are negatively charged at pH 7 or higher, and in some cases at pH 6, 4, or even 2. Specific examples of anionic polymers include poly(carboxylic acids), such as poly(acrylic acid) or poly(methacrylic acid); anionic polysaccharides including alginate, hyaluronic acid, pectin, agar, carrageenan, gellan gum, gum arabic, guar gum, xanthan gum, and carboxymethyl cellulose; sulfonate polymers such as poly(2-acrylamido-2-methylpropanesulfonate) (polyAMPS) or polystyrene sulfonate; and polyphosphates. Anionic polymers may be provided in the form of salts, such as sodium or potassium salts.

[0118] In some aspects of this disclosure, a system is provided configured to dispense an amino acid- or amino alcohol-based polyamine compound and a polymer crosslinked with the amino acid- or amino alcohol-based polyamine compound under conditions that cause the polyamine compound and the polymer to crosslink with each other. Such a system can be used to form crosslinked hydrogels in vivo or in vitro.

[0119] In some embodiments, a system is provided configured to dispense an amino acid- or amino alcohol-based polyamine compound and a reactive polymer, said reactive polymer comprising a plurality of reactive groups covalently crosslinked with the primary amine group of the amino acid- or amino alcohol-based polyamine compound under conditions that cause the amino acid- or amino alcohol-based polyamine compound and the reactive polymer to covalently crosslink with each other. Exemplary amino acid- and amino alcohol-based polyamine compounds and exemplary reactive polymers are as described above, wherein the reactive polymer comprises a reactive multi-arm polymer containing a plurality of polymer arms, wherein two or more polymer arms of the multi-arm polymer contain one or more reactive end groups, such as cyclic imide ester groups. In some embodiments, those conditions include an environment having an alkaline pH, for example, a pH range of about 9 to about 11.

[0120] In some embodiments, a system is provided comprising (a) a first composition comprising a polyamine compound based on an amino acid or an amino alcohol, and (b) a second composition comprising a reactive polymer that forms a covalently crosslinked polymer with the polyamine compound based on an amino acid or an amino alcohol. In some embodiments, a third composition in the form of an accelerator composition is also provided.

[0121] The first composition may be a first fluid composition comprising a polyamine compound based on an amino acid or an amino alcohol, or a first dry composition comprising a polyamine compound based on an amino acid or an amino alcohol. A suitable fluid, such as water for injection, a saline solution, etc., may be added to the first dry composition to form the first fluid composition. In addition to the polyamine compound based on an amino acid or an amino alcohol, the first composition may also contain additives, including those described below. The first composition may be provided in a suitable reservoir, such as a syringe, vial, or other reservoir.

[0122] The second composition may be a second fluid composition comprising a reactive multi-arm polymer, or a second dry composition comprising a reactive multi-arm polymer. A suitable fluid, such as water for injection, a saline solution, etc., may be added to the second dry composition to form the second fluid composition. In addition to the reactive multi-arm polymer, the second composition may also contain additives, including those described below. The second composition may be provided in a suitable reservoir, such as a syringe, vial, or other reservoir.

[0123] The accelerator composition may be a fluid accelerator composition buffered to an alkaline pH, or a dry composition containing an alkaline buffer composition. A suitable fluid, such as water for injection, a saline solution, etc., may be added to the dry composition to form a fluid accelerator composition buffered to an alkaline pH. The accelerator composition may also contain additives, including those described below. The accelerator composition may be provided in a suitable reservoir, such as a syringe, vial, or other reservoir.

[0124] In some embodiments, the amino acid- or amino alcohol-based polyamine compound is initially mixed with a reactive multi-arm polymer at an acidic pH, where covalent cross-linking between the reactive groups of the reactive multi-arm polymer and the primary amine groups of the amino acid- or amino alcohol-based polyamine compound is suppressed. Then, when covalent cross-linking is required, the pH of the mixture of the amino acid- or amino alcohol-based polyamine compound and the reactive multi-arm polymer is changed from acidic to alkaline, resulting in covalent cross-linking between them.

[0125] In some embodiments, a system is provided in which a precursor fluid composition comprising an amino acid or amino alcohol-based polyamine compound and a reactive multi-arm polymer is prepared and buffered to an acidic pH. For example, the pH range of the precursor fluid composition may be, for example, from about 3 to about 5, etc. In some embodiments, the precursor fluid composition contains an acid buffer, such as sodium dihydrogen phosphate. The precursor fluid composition may also contain additives, including those described below.

[0126] The system also includes a fluid accelerator composition containing an alkaline buffer. For example, the fluid accelerator may contain an alkaline buffer that provides the fluid accelerator with a pH range of, for example, from about 9 to about 11. In some embodiments, the accelerator composition may contain sodium borate and disodium hydrogen phosphate, etc. The fluid accelerator composition may also contain additives, including those described below.

[0127] Precursor fluid compositions buffered to acidic pH and containing polyamine compounds based on amino acids or amino alcohols and reactive multi-arm polymers, and fluid accelerator compositions buffered to alkaline pH, can be combined in vivo or in vitro to form covalently cross-linked hydrogels.

[0128] In other embodiments, a system is provided configured to dispense an amino acid- or amino alcohol-based polyamine compound as described herein and an anionic polymer comprising a plurality of anionic groups, the anionic groups being crosslinked with the cationic primary amino groups of the amino acid- or amino alcohol-based polyamine compound under pH conditions that cause the amino acid- or amino alcohol-based polyamine compound and the anionic polymer to be ionicly crosslinked with each other. Exemplary amino acid- and amino alcohol-based polyamine compounds and anionic polymers are as described above.

[0129] In some embodiments, a system is provided comprising (a) a first composition comprising a polyamine compound based on an amino acid or an amino alcohol, and (b) a second composition comprising an anionic polymer that forms an ionic crosslink with the polyamine compound based on an amino acid or an amino alcohol. In some embodiments, a third composition in the form of an accelerator composition is provided.

[0130] The first composition may be a first fluid composition comprising a polyamine compound based on an amino acid or an amino alcohol, or a first dry composition comprising a polyamine compound based on an amino acid or an amino alcohol. A suitable fluid, such as water for injection, a saline solution, etc., may be added to the first dry composition to form the first fluid composition. In addition to the polyamine compound based on an amino acid or an amino alcohol, the first composition may also contain additives, including those described below. The first composition may be provided in a suitable reservoir, such as a syringe, vial, or other reservoir.

[0131] The second composition may be a second fluid composition comprising an anionic polymer, or a second dry composition comprising an anionic polymer. A suitable fluid, such as water for injection, a saline solution, etc., may be added to the second dry composition to form the second fluid composition. In addition to the anionic polymer, the second composition may also contain additives, including those described below. The second composition may be provided in a suitable reservoir, such as a syringe, vial, or other reservoir.

[0132] The accelerator composition may be a fluid accelerator composition buffered to an alkaline or acidic pH, or a dry composition containing an alkaline or acidic buffer composition. A suitable fluid, such as water for injection, a saline solution, etc., may be added to the dry composition to form a fluid accelerator composition buffered to an alkaline or acidic pH. The accelerator composition may also contain additives, including those described below. The accelerator composition may be provided in a suitable reservoir, such as a syringe, vial, or other reservoir.

[0133] In some embodiments, the amino acid- or amino alcohol-based polyamine compound is initially mixed with the anionic polymer at an alkaline pH where the primary amine group of the amino acid- or amino alcohol-based polyamine compound carries a neutral charge, or at an acidic pH where the carboxyl group of the anionic polymer carries a neutral charge, thereby inhibiting ionic crosslinking. Then, when crosslinking is required, the pH of the mixture of the amino acid- or amino alcohol-based polyamine compound and the anionic polymer is changed from the acidic or alkaline pH to a more neutral pH, resulting in ionic crosslinking between the amino acid- or amino alcohol-based polyamine compound and the anionic polymer.

[0134] In some embodiments, a system is provided in which a precursor fluid composition is prepared, the precursor fluid composition being buffered to an acidic or alkaline pH, and comprising an amino acid or amino alcohol-based polyamine compound and an anionic polymer. The precursor fluid composition may also contain additives, including those described below.

[0135] The system also includes a fluid accelerator composition buffered to an alkaline or acidic pH. The amount and type of buffer in the fluid accelerator composition are selected such that, when mixed with a precursor fluid composition containing an amino acid- or amino alcohol-based polyamine compound and an anionic polymer, the resulting mixture has a more neutral pH at which the amino acid- or amino alcohol-based polyamine compound carries a positive charge and the anionic polymer carries a negative charge, thereby forming ionic crosslinks between them. The fluid accelerator composition may also contain additives, such as those described below.

[0136] Precursor fluid compositions buffered to acidic or alkaline pH and containing polyamine compounds based on amino acids or amino alcohols and anionic polymers, and fluid accelerator compositions buffered to alkaline or acidic pH, can be combined in vivo or in vitro to form ionic crosslinked hydrogels.

[0137] Examples of additives used in the above compositions include therapeutic agents such as anti-angiogenic agents, cytotoxic agents, chemotherapeutic agents, checkpoint inhibitors, immunomodulatory cytokines, T-cell agonists, and STING (interferon gene stimulator) agonists.

[0138] Examples of additives also include imaging agents other than any iodine that may be present in radiopaque products. Such imaging agents include (a) fluorescent dyes, such as fluorescein, indocyanine green, or fluorescent proteins (e.g., green, blue, and blue-green fluorescent proteins); (b) contrast agents used in conjunction with magnetic resonance imaging (MRI), including contrast agents containing elements that form paramagnetic ions, such as Gd(III), Mn(II), Fe(III), and compounds containing them (including chelates), such as gadolinium ions chelated with diethylenetriaminepentaacetic acid; (c) contrast agents used in conjunction with ultrasound imaging, including organic and inorganic echogenic particles (i.e., particles that increase reflected ultrasound energy) or organic and inorganic echolucent particles (i.e., particles that decrease reflected ultrasound energy); and (d) radioactive contrast agents, such as those based on the clinically important isotope 99mTc, and other gamma emitters such as 123I, 125I, etc. (e) Positron emitters, such as 18F, 11C, 13N, 15O, and 68Ga, can be used to produce functionalized radioactive capping layers; (f) radioactive contrast agents, such as particles of tantalum, tungsten, rhenium, niobium, molybdenum, and their alloys, wherein the metal particles can be spherical or non-spherical; non-ionic radioactive contrast agents, such as iohexol, iodixanol, iopromide, iopamidol, ioxilan, or iopromide; ionic radioactive contrast agents, such as diatrizoate, iothalamate, metrizoate, or ioxaglate; and iodized oils, including ethyl iodized poppy oil. (g) poppyseed oil (available as Lipiodol®), and (g) a contrast agent for use in conjunction with near-infrared (NIR) imaging, which may be selectively used to impart near-infrared fluorescence to the cover layer of this disclosure, thereby allowing deep tissue imaging and device labeling, such as NIR-sensitive nanoparticles, such as gold nanoshells, carbon nanotubes (e.g., nanotubes derived with hydroxyl or carboxyl groups, such as partially oxidized carbon nanotubes), dye-containing nanoparticles, such as dye-doped nanofibers and dye-encapsulated nanoparticles, and semiconductor quantum dots, etc. NIR-sensitive dyes include cyanine dyes, squaric acid cyanine, phthalocyanine, porphyrin derivatives, and borobispyrrolidone (BODIPY) analogues, etc.

[0139] In various embodiments, a system is provided that includes one or more delivery devices for delivering the compositions described herein to a subject.

[0140] In some embodiments, a delivery device is provided comprising (a) a first reservoir containing a first fluid composition comprising a polyamine compound based on an amino acid or amino alcohol as described above, and (b) a second reservoir containing a second fluid composition comprising a reactive multi-arm polymer as described above.

[0141] In some embodiments, a delivery device is provided comprising (a) a first reservoir containing a first fluid composition comprising, as described above, an amino acid or amino alcohol-based polyamine compound and a reactive multi-arm polymer as described above, wherein the first fluid composition is buffered to an acidic pH to inhibit covalent crosslinking, as described above, a precursor fluid composition, and (b) a second reservoir containing a second fluid composition, as described above, a fluid accelerator composition.

[0142] In some embodiments, a delivery device is provided comprising (a) a first reservoir containing a first fluid composition comprising a polyamine compound based on an amino acid or amino alcohol as described above, and (b) a second reservoir containing a second fluid composition comprising an anionic polymer as described above.

[0143] In some embodiments, a delivery device is provided comprising (a) a first reservoir containing a first fluid composition comprising a polyamine compound based on an amino acid or amino alcohol as described above and an anionic polymer as described above, the first fluid composition being buffered to an acidic or alkaline pH to inhibit ionic crosslinking, such as the precursor fluid composition described above, and (b) a second reservoir containing a second fluid composition, such as the fluid accelerator composition buffered to an alkaline or acidic pH described above.

[0144] In each of the above cases, during operation, the first composition and the second composition are dispensed from the first reservoir and the second reservoir and combined, thereby crosslinking the polyamine compound based on amino acids or amino alcohols and the reactive multi-arm polymer to form a hydrogel.

[0145] In a particular implementation, reference Figure 9The system may include a delivery device 910 comprising a dual-tube syringe, the syringe including a first tube 912a having a first tube outlet 914a (the first tube containing a first fluid composition), a first plunger 916a movable within the first tube 912a, a second tube 912b having a second tube outlet 914b (the second tube 912b containing a second fluid composition), and a second plunger 916b movable within the second tube 912b. In some embodiments, the device 910 may further include a mixing section 918 having a first mixing section inlet 918a1 in fluid communication with the first tube outlet 914a, a second mixing section inlet 918b1 in fluid communication with the second tube outlet, and a mixing section outlet 918o. Also shown is a syringe holder 922 configured to hold the first syringe tube 912a and the second syringe tube 912b in a fixed relationship, and a plunger cap 924 configured to hold the first plunger 916a and the second plunger 916b in a fixed relationship.

[0146] In some embodiments, the delivery device may also include a cannula or conduit configured to receive a first fluid composition and a second fluid composition from a first tube and a second tube. For example, the cannula or conduit may be configured to form a fluid connection with the outlet of the mixing section by attaching the cannula or conduit to the outlet of the mixing section (e.g., via a suitable fluid connector such as a Luer connector).

[0147] As another example, the catheter may be a multi-lumen catheter including a first lumen and a second lumen, the proximal end of the first lumen being configured to form a fluid connection with a first tube outlet, and the proximal end of the second lumen being configured to form a fluid connection with a second tube outlet. In some embodiments, the multi-lumen catheter may include a mixing portion having a first mixing portion inlet in fluid communication with the distal end of the first lumen, a second mixing portion inlet in fluid communication with the distal end of the second lumen, and a mixing portion outlet.

[0148] During operation, when the first and second plungers are depressed, the first and second fluid compositions are dispensed from the first and second tubes, whereby they mix and ultimately crosslink to form a crosslinked hydrogel, which is then applied to or into the subject's tissue. For example, the first and second fluid compositions may enter a mixing section from the first and second tubes via a first and second mixing section inlet, whereby they mix to form a mixture that exits the mixing section via a mixing section outlet. In some embodiments, a cannula or catheter is attached to the mixing section outlet, allowing the mixture to be applied to the subject after passing through the cannula or catheter.

[0149] As another example, a first fluid composition may enter a first lumen of the multi-lumen catheter from a first tube outlet, and a second fluid composition may enter a second lumen of the multi-lumen catheter from a second tube outlet. In some embodiments, the first fluid composition and the second fluid composition may enter a mixing section at the distal end of the multi-lumen catheter from the first lumen and the second lumen, respectively, via a first mixing section inlet and a second mixing section inlet, whereby the first fluid composition and the second fluid composition mix to form a mixture, which exits the mixing section via a mixing section outlet.

[0150] Regardless of the type of apparatus used to mix the first and second fluid compositions, or how the first and second fluid compositions are mixed, after the mixture of the first and second fluid compositions is formed, the mixture is initially in a fluid state and can be applied to a subject (e.g., a mammal, particularly a human) by various techniques. Alternatively, the first and second fluid compositions can be applied to the subject independently, and the fluid mixture of the first and second fluid compositions is formed inside or on the surface of the subject. In either method, the fluid mixture of the first and second fluid compositions is formed and used in various medical procedures.

[0151] For example, in applications such as the treatment of diseases and cancers, as well as tissue repair and regeneration, the first fluid composition and the second fluid composition, or a fluid mixture thereof, can be injected to provide spacers between tissues; the first fluid composition and the second fluid composition, or a fluid mixture thereof, can be injected (e.g., in the form of blisters) to provide reference markers; the first fluid composition and the second fluid composition, or a fluid mixture thereof, can be injected for tissue enlargement or regeneration; the first fluid composition and the second fluid composition, or a fluid mixture thereof, can be injected as fillers or replacements for soft tissues; the first fluid composition and the second fluid composition, or a fluid mixture thereof, can be injected to provide mechanical support for damaged tissues; the first fluid composition and the second fluid composition, or a fluid mixture thereof, can be injected as scaffolds; the first fluid composition and the second fluid composition, or a fluid mixture thereof, can be injected as embolic compositions; the first fluid composition and the second fluid composition, or a fluid mixture thereof, can be injected as lifting agents for the removal of internal cysts; and / or the first fluid composition and the second fluid composition, or a fluid mixture thereof, can be injected as carriers of therapeutic agents. The first fluid composition and the second fluid composition, or a fluid mixture thereof, can also be injected into the left atrial appendage during left atrial appendage occlusion surgery. In some implementations, the first fluid composition and the second fluid composition or a fluid mixture thereof may be injected into the left atrial appendage after the introduction of an occlusion device (such as the Watchman® left atrial appendage occlusion device available from Boston Scientific Corporation).

[0152] Upon application of the compositions disclosed herein (either as a first fluid composition and a second fluid composition mixed in vivo, or as a fluid mixture of the first fluid composition and the second fluid composition), a cross-linked hydrogel is ultimately formed at the application site.

[0153] After application, the compositions disclosed herein can be imaged using suitable imaging techniques such as ultrasound-based or X-ray-based imaging techniques such as computed tomography or X-ray fluorescence fluoroscopy.

[0154] As can be seen from the above, the compositions disclosed herein can be used in various medical procedures, including: procedures for implanting a reference marker comprising a crosslinked product of a first fluid composition and a second fluid composition; procedures for implanting a tissue regeneration scaffold comprising a crosslinked product of a first fluid composition and a second fluid composition; procedures for implanting a tissue support comprising a crosslinked product of a first fluid composition and a second fluid composition; procedures for implanting a tissue expander comprising a crosslinked product of a first fluid composition and a second fluid composition; procedures for implanting an embolization composition comprising a crosslinked product of a first fluid composition and a second fluid composition; procedures for implanting a lifting agent comprising a crosslinked product of a first fluid composition and a second fluid composition; procedures for introducing a left atrial appendage occlusion composition comprising a crosslinked product of a first fluid composition and a second fluid composition; procedures for implanting a therapeutic agent-containing depot comprising a crosslinked product of a first fluid composition and a second fluid composition; procedures for tissue enlargement comprising implanting a crosslinked product of a first fluid composition and a second fluid composition; and procedures for introducing a crosslinked product of a first fluid composition and a second fluid composition between a first tissue and a second tissue to separate the first tissue from the second tissue.

[0155] The first fluid composition and the second fluid composition, a fluid mixture of the first fluid composition and the second fluid composition, or a crosslinked product of the first fluid composition and the second fluid composition can be injected in combination with a variety of medical procedures including: injection for septal use between the prostate or vagina and rectum in radiotherapy for rectal cancer; injection for septal use between the rectum and prostate in radiotherapy for prostate cancer; subcutaneous injection for palliative treatment of prostate cancer; transurethral or submucosal injection for female stress urinary incontinence; intravesical injection for urinary incontinence; intrauterine injection for Asherman's syndrome; submucosal injection for anal incontinence; percutaneous injection for heart failure; intramyocardial injection for heart failure and dilated cardiomyopathy; transendocardial injection for myocardial infarction; intra-articular injection for osteoarthritis; and injections for spinal fusion and spinal, oral and maxillofacial, and orthopedic procedures. Spinal injections for traumatic surgery; spinal injections for posterolateral lumbar fusion; intradiscal injections for degenerative disc diseases; injections between the pancreas and duodenum for pancreatic cancer imaging; resection bed injections for oropharyngeal cancer imaging; peritumoral injections for bladder cancer imaging; submucosal injections for gastrointestinal tumors and polyps; visceral pleural injections for lung biopsies; renal injections for type 2 diabetes and chronic kidney disease; renal cortical injections for chronic kidney disease with congenital abnormalities from the kidneys and urethra; intravitreal injections for neovascular age-related macular degeneration; intratympanic injections for sensorineural hearing loss; and dermal injections to correct wrinkles, creases and folds, signs of facial fat loss, volume reduction, superficial to deep contour defects, correction of depressed skin scars, perioral wrinkles, lip augmentation, facial fat atrophy, and stimulate natural collagen production.

[0156] The cross-linked hydrogel compositions according to this disclosure include smooth compositions for medical applications, compositions for the release of therapeutic agents (e.g., by including one or more therapeutic agents in the matrix of the cross-linked hydrogel), and implants (which can be formed in vitro or in vivo) (e.g., compositions used as tissue markers, compositions acting as spacers to reduce the side effects of off-target radiation therapy, cosmetic compositions, etc.).

[0157] It should be understood that this disclosure is merely illustrative in many respects and that changes in detail may be made without departing from the scope of this disclosure. To the appropriate extent, this may include the use of any feature of one embodiment in other embodiments.

Claims

1. An amino acid-based polyamine compound formed by: (a) a ring-opening reaction of two or more amino acid N-carboxyl anhydride molecules with a polyol having two or more hydroxyl groups under acid catalysis, or (b) an ester coupling reaction of two or more amine-protected amino acid molecules with a polyol having two or more hydroxyl groups in the presence of an ester coupling agent.

2. The amino acid-based polyamine compound according to claim 1, wherein the amino acid N-carboxyl ring anhydride molecule is selected from glycine N-carboxyl ring anhydride, lysine N-carboxyl ring anhydride, ornithine N-carboxyl ring anhydride and cysteine ​​N-carboxyl ring anhydride.

3. The amino acid-based polyamine compound according to claim 1, wherein the amine-protected amino acid is selected from amine-protected glycine, amine-protected lysine, amine-protected ornithine, and amine-protected cysteine.

4. An amino acid-based polyamine compound comprising (a) a polyol residue having two or more hydroxyl groups, and (b) two or more monoamine-terminated amino acid residues, each of the monoamine-terminated amino acid residues being linked by an ester bond at one of the two or more hydroxyl group residue sites.

5. The amino acid-based polyamine compound according to claim 4, wherein the amine-terminated amino acid residue is selected from amine-terminated glycine residues, amine-terminated lysine residues, amine-terminated ornithine residues, and amine-terminated cysteine ​​residues.

6. The amino acid-based polyamine compound according to any one of claims 1-5, wherein the polyol has 3 to 20 hydroxyl groups.

7. An amino alcohol-based polyamine compound formed by (a) an amino alcohol molecule protected by an amine having one or more amino groups and a single hydroxyl group and (b) a polycarboxylic acid compound having two or more carboxyl groups in the presence of an ester coupling agent.

8. The polyamine compound based on an amino alcohol according to claim 7, wherein the amine-protected amino alcohol is an amine-protected C2-C having one, two, three, or four amino groups and a single hydroxyl group. 10 -Amino alcohol.

9. A polyamine compound based on an amino alcohol, comprising (a) residues of a polycarboxylic acid compound having two or more carboxyl groups, and (b) two or more monoamine-terminated amino alcohol residues, each of the monoamine-terminated amino alcohol residues having one or more amino groups and being linked by an ester bond at one residue site of the two or more carboxyl groups.

10. The amino alcohol-based polyamine compound according to claim 9, wherein the amine-terminated amino alcohol residue is an amine-terminated C2-C compound having one, two, three, or four amino groups. 10 -Amino alcohol residues.

11. The amino alcohol-based polyamine compound according to any one of claims 7-10, wherein the polycarboxylic acid has 3 to 20 carboxyl groups.

12. A system for forming a hydrogel comprising a polyamine compound based on an amino acid or amino alcohol according to any one of claims 1-11 and a polymer crosslinked with the amino acid or amino alcohol-based polyamine compound.

13. The system of claim 12, wherein the polymer that crosslinks with the amino acid- or amino alcohol-based polyamine compound is a reactive multi-arm polymer comprising a plurality of hydrophilic polymer arms, the hydrophilic polymer arms comprising hydrophilic polymer segments and reactive end groups, the reactive end groups being covalently crosslinked with the primary amine group of the amino acid- or amino alcohol-based polyamine compound.

14. The system of claim 13, wherein the hydrophilic polymer segment is selected from polyepoxide segments, polyester segments, polyoxazoline segments, polydioxane-hexanone segments, and polypeptide segments, and / or wherein the reactive group is an electrophilic group selected from imidazole esters, imidazole carboxylic esters, benzotriazole esters, or imide esters.

15. The system of claim 13, wherein the polymer crosslinked with the amino acid- or amino alcohol-based polyamine compound is an anionic polymer comprising a plurality of anionic groups crosslinked with a plurality of primary amine ions of the amino acid- or amino alcohol-based polyamine compound.

Images