An alkoxysilane-PEG crosslinking agent, a physical / chemical double crosslinking hydrogel and application thereof

Abstract

The application belongs to the technical field of medical polymer materials, and particularly relates to an alkoxy silane PEG crosslinking agent, a physical / chemical double crosslinking hydrogel and application thereof. The application utilizes a small molecule alkoxy silane which is insoluble in water to modify the end of a PEG macromolecule, and after forming a homogeneous system with a physical hydrogel, the alkoxy silane is slowly hydrolyzed and crosslinked, and then a physical / chemical double crosslinking hydrogel is formed. Moreover, the physical / chemical double crosslinking hydrogel disclosed in the application combines the advantages of a physical hydrogel and a chemical hydrogel: the water system before crosslinking has good temperature sensitivity and shear thinning characteristics, and after mixing various functional additives at low temperature, the water system can be delivered through a medical needle tube and a catheter; the physical crosslinking thermally induced gel is in situ gelled at an application site and is beneficial to subsequent chemical crosslinking; after being injected into a target site, the chemical crosslinking reaction gradually occurs, and after complete crosslinking, the mechanical properties and stability of the hydrogel are greatly improved.

Description

Technical Field

[0001] This invention belongs to the field of medical polymer materials technology, specifically relating to an alkoxysilane-modified PEG crosslinking agent, a physical / chemical double crosslinked hydrogel, and their applications. Background Technology

[0002] Hydrogels are a class of materials with a three-dimensional polymer network containing a large amount of water. Hydrogels have a structure similar to the extracellular matrix and their physical and chemical properties are highly tunable, attracting widespread attention in the biomedical field, including applications such as vascular embolization, tissue engineering, cell encapsulation, and wound healing. Hydrogels can be classified into physical hydrogels and chemical hydrogels based on the nature of the three-dimensional network they form. The interactions that form physical hydrogels mainly include hydrogen bonding, hydrophobic interactions, electrostatic interactions, physical entanglement of polymer chains, and crystallization. The advantage of physically cross-linked hydrogels lies in their excellent self-healing and self-repairing properties, stemming from the reversible breakage and recovery of the physical interactions that form them. Chemical hydrogels contain covalent bonds between different polymer chains. Methods for preparing chemically cross-linked hydrogels mainly include condensation reactions, free radical polymerization, enzymatic cross-linking, and high-energy radiation cross-linking.

[0003] In the biomedical field, chemically cross-linked hydrogels typically possess good mechanical properties and long degradation times. However, their application in living organisms is limited by challenges such as high synthesis difficulty and stringent cross-linking conditions.

[0004] Therefore, designing a physical / chemical double crosslinked hydrogel with controllable crosslinking time, mild crosslinking conditions, and delivery via injection or other methods has significant application value. Summary of the Invention

[0005] In view of this, the first objective of the present invention is to provide a hydrogel crosslinking agent that can regulate the rate of crosslinking reaction, addressing the problems existing in the prior art.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] An alkoxysilane-modified PEG crosslinking agent, the structure of which is a PEG polymer modified with alkoxysilyl crosslinking groups at the ends.

[0008] Among them, alkoxysilaneized PEG crosslinking agents are obtained by reacting the functional groups at the ends of different PEG polymers with alkoxysilanes containing different functional groups, such as the reaction of the hydroxyl or amino groups at the ends of PEG with the isocyanate groups in alkoxysilanes.

[0009] It is worth noting that alkoxysilanes are small molecule compounds that are insoluble in water. When modified at the end of a PEG macromolecule, they can form a homogeneous system with a physical hydrogel, followed by slow hydrolysis and cross-linking to form a double-crosslinked hydrogel. However, a simple aqueous solution of alkoxysilane-modified PEG cross-linking agent will rapidly drain after being injected subcutaneously. The alkoxysilane-modified PEG cross-linking agent disclosed in this invention can prevent its rapid loss at the injection site by preparing a physical / chemical double hydrogel.

[0010] Furthermore, the molecular weight of the PEG polymer ranges from 400 to 100,000, including one or more of linear PEG, 4-arm PEG, 6-arm PEG, and 8-arm PEG; the alkoxysilyl crosslinking groups modified at the ends of the polymer include one or more of trimethoxysilyl, dimethoxymethylsilyl, triethoxysilyl, diethoxymethylsilyl, triphenoxysilyl, diphenoxymethylsilyl, triacetoxysilyl, and diacetoxymethylsilyl.

[0011] It is worth noting that the crosslinking process of alkoxysilane-modified PEG crosslinking agents mainly consists of two steps: first, the alkoxysilane groups undergo hydrolysis to generate silanol groups; then, the silanol groups and alkoxysilanes or other hydrolyzed silanol groups undergo condensation to form siloxane bonds. The entire reaction is accompanied by the formation of small molecule alcohols. This crosslinking process can occur spontaneously and slowly in neutral aqueous solutions. The rate of the crosslinking reaction can be effectively controlled by adjusting the temperature and pH of the entire system. Within the range of 0-40℃, the higher the temperature, the faster the crosslinking reaction. At neutral pH, the hydrolysis and condensation reactions are relatively slow; at pH 7-10, the stronger the alkalinity, the faster the hydrolysis and condensation reactions. Therefore, depending on the crosslinking conditions of the system, the crosslinking time can range from a few minutes to several days.

[0012] The second objective of this invention is to address the problems existing in the prior art by providing a physical / chemical double crosslinked hydrogel with controllable crosslinking time, mild crosslinking conditions, and simple delivery method.

[0013] A physical / chemical double crosslinked hydrogel comprises the following raw materials in parts by weight: 6-13 parts of amphiphilic polymer, 1-10 parts of alkoxysilaneized PEG crosslinking agent as described above, 0-30 parts of functional additives, and 47-93 parts of water-based solvent.

[0014] It is worth noting that the hydrogel with a physical / chemical dual crosslinking structure proposed in this invention is an aqueous system obtained by blending an amphiphilic polymer, an alkoxysilane-modified PEG crosslinking agent, and an aqueous solvent. This aqueous system is a free-flowing sol at low temperatures, which can be easily mixed with various functional additives, thus endowing the dual crosslinked hydrogel with the potential for various medical applications. As the temperature rises, the aqueous system undergoes a sol-gel phase transition, thereby forming the first physical network of the hydrogel. The forces that form this network are physical interactions, mainly including hydrogen bonds that can be reversibly broken and restored, hydrophobic interactions, etc. The thermogenic gelation ability and physical network of the aqueous system make it easier to be delivered through needles and catheters used in clinical applications, thus adapting to various in vivo applications. After the hydrogel is retained at the target site, the second chemical crosslinking network gradually forms. The alkoxysilyl groups modified at the end of the alkoxysilane-modified PEG crosslinking agent hydrolyze to form silanols and small molecule alcohols. The silanols can undergo condensation reactions with the alkoxysilyl groups or other hydrolyzed silanol groups to form silyl ether crosslinking bonds. The more crosslinking bonds formed and the higher the degree of crosslinking, the more stable the morphology of the hydrogel and the better its mechanical properties. The most important advantage of the second-layer chemical cross-linking network is that it improves the mechanical properties of the hydrogel, thus extending the stability and retention time of the hydrogel in various application scenarios (in vivo, culture flasks, etc.).

[0015] Furthermore, the amphiphilic polymer comprises 40wt% to 60wt% of hydrophilic blocks and 60wt% to 40wt% of hydrophobic blocks of polyamino acids.

[0016] Furthermore, the hydrophilic block is polyethylene glycol with a molecular weight range of 750 to 5000 or polyvinylpyrrolidone with a molecular weight range of 750 to 5000; the hydrophobic block is polyamino acid with a molecular weight range of 400 to 5000.

[0017] Furthermore, the polyamino acid includes L or D -Alanine, L or D -Phenylalanine, L or D -Leucine, L or D -Isoleucine, L or D -valine, L or D -Lysine, L or D -Aspartic acid, L or D -Aspartic acid methyl ester L or D - Ethyl aspartate L or D-Aspartic acid benzyl ester L or D -Glutamic acid, L or D - glutamate methyl ester L or D -Ethyl glutamate, L or D Homopolymers of β-benzyl glutamate, or the above L Type or D A copolymer formed from monomers in any form.

[0018] Furthermore, the amphiphilic polymer is selected from one or more of the following: AB diblock copolymer, BAB triblock copolymer, AgB or BgA type grafted block copolymer, n-arm-(AB) or n-arm-(BA) type N-arm block copolymer, wherein n is an integer greater than 2 and less than 10.

[0019] It is worth noting that the amphiphilic polymer disclosed in this invention uses polyethylene glycol (PEG) or polyvinylpyrrolidone (PVP) as the hydrophilic segment and polyamino acid as the hydrophobic segment. This amphiphilic polymer can be dissolved in an aqueous solvent to prepare an aqueous system with sol-gel phase transition capability. At the same time, the aqueous system has shear thinning capability, so it can be better delivered to the designated site through a needle or catheter. After injection into the target site, it can better maintain the shape of the hydrogel, which is conducive to the formation of further chemical cross-linking network.

[0020] In particular, the physical / chemical dual crosslinked hydrogel disclosed in this invention combines the advantages of physical hydrogels and chemical hydrogels. Before chemical crosslinking, it has shear-thinning properties, which allows for better injection through needles and catheters, while after chemical crosslinking, it has better mechanical properties.

[0021] Furthermore, the functional additives include:

[0022] X-ray contrast agents: one or more combinations of iodized oil, 2-octyl triiodobenzoate, octyl triiodobenzoate, iohexol, iopamidol, iodixanol, iofluoxetine, iodamide, and iopromide; or,

[0023] Fluorescent dyes: one or more combinations of Rhodamine, Nile Red, Cy3, Cy5, Cy5.5, Cy7, and tetraphenylethylene; or,

[0024] Magnetic resonance imaging contrast agents: gadopentetate dimeglumine, gadofusamide, gadobutrol, superparamagnetic iron oxide particles; or,

[0025] Anticancer drugs: one or more of the following: doxorubicin, paclitaxel, docetaxel, vinorelbine, gemcitabine, 5-fluorouracil, capecitabine, sorafenib, lenvatinib, ramucirumab, apatinib, cisplatin, oxaliplatin, bleomycin, regorafenib, cabozantinib, irinotecan, and nivolumab; or,

[0026] Diabetes medications: one or more combinations of insulin, metformin, liximab, liraglutide, and exenatide; or,

[0027] Antibacterial drugs: one or more of penicillin and gentamicin, or a combination thereof.

[0028] It is worth noting that the physically / chemically cross-linked hydrogel disclosed in this invention has good encapsulation capacity. The amphiphilic polymer can self-assemble into micelles in aqueous solutions, effectively encapsulating hydrophobic drugs. It can also interact with protein drugs through hydrogen bonding and hydrophobic interactions, achieving a good sustained-release effect. Encapsulating functional additives with different properties expands the hydrogel's application potential in various fields. Encapsulating X-ray contrast agents can endow the hydrogel with X-ray imaging capabilities, making it suitable for use as a vascular embolization agent and a soft tissue marker for tumor radiotherapy; encapsulating fluorescent dyes can endow the hydrogel with fluorescence tracking capabilities; it can also encapsulate drugs such as anticancer drugs, diabetes drugs, and antibiotics.

[0029] Furthermore, the water-based solvent is one or more of pure water, physiological saline, phosphate buffer solution, cell culture medium, and tissue culture medium.

[0030] It should be understood that although the present invention limits the types of solvents, other aqueous solutions or media not based on organic solvents can be used as solvents in appropriate circumstances.

[0031] Considering the initial viscosity and other properties of the physical / chemical dual crosslinked hydrogel, the physical / chemical dual crosslinked hydrogel disclosed in this invention further includes 0 to 15 parts by weight of a modifier, which includes one or more of the following: nano-lithium diatomaceous earth, xylitol, chitin, sorbitol, mannitol, Tween 20, Tween 60, Tween 80, carbomer, polyethylene glycol, sodium carboxymethyl cellulose, chondroitin, gelatin, and protein glue.

[0032] The third objective of this invention is to provide an application of the physical / chemical dual crosslinked hydrogel described above in the fields of vascular embolization, tissue repair, cell encapsulation, and wound healing.

[0033] Compared with existing technologies, this invention utilizes water-insoluble small-molecule alkoxysilanes to modify the ends of PEG macromolecules, forming a homogeneous system with a physical hydrogel, followed by slow hydrolysis and cross-linking, thereby forming a physical / chemical dual-crosslinked hydrogel. In particular, in existing technologies, the aqueous solution of a simple alkoxysilane-modified PEG crosslinking agent injected subcutaneously will rapidly drain away. The alkoxysilane-modified PEG crosslinking agent disclosed in this invention, through the preparation of a physical / chemical dual hydrogel, can prevent its rapid loss at the injection site in vivo.

[0034] Furthermore, the physical / chemical dual crosslinked hydrogel disclosed in this invention combines the advantages of both physical and chemical hydrogels: the aqueous system before chemical crosslinking exhibits excellent temperature-sensitive and shear-thinning properties, and can be delivered via medical needles and catheters after mixing with various functional additives at low temperatures; the thermotropic gel formed in situ at the application site through physical crosslinking, which is beneficial for subsequent chemical crosslinking; the chemical crosslinking reaction gradually occurs after injection into the target site, and the crosslinking process of the alkoxysilane crosslinking agent consists of two steps: hydrolysis and condensation, which makes the crosslinking process more stable and controllable. After complete crosslinking, the mechanical properties and stability of the hydrogel are greatly improved; in addition, the addition of functional additives further expands the application range of the physical / chemical dual crosslinked hydrogel. Attached Figure Description

[0035] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are merely embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0036] Figure 1 The above is the 1H NMR spectrum of the amphiphilic polymer mPEG-PAF (coploymer-6) disclosed in Example 14 of this invention;

[0037] Figure 2 The alkoxysilanized PEG crosslinking agent 4-arm-PEG prepared in Example 4 of this invention 5000 -IPTS (crosslinking agent-4) 1H NMR spectrum;

[0038] Figure 3 The 5wt% 4-arm-PEG disclosed in Example 36 of this invention 5000 Crosslinking time of IPTS (crosslinking agent-4) PBS solution at different pH levels and at different temperatures at pH 10;

[0039] Figure 4The 10wt% PA-PEG-PA (coploymer-4) and 5wt% 4-arm-PEG disclosed in Example 27 of this invention 5000 - Shear thinning performance test of IPTS (crosslinking agent-4) water system after 0h and 4h at 25℃;

[0040] Figure 5 The 10wt% PA-PEG-PA (coploymer-4) and 5wt% 4-arm-PEG disclosed in Example 38 of this invention are examples of such materials. 5000 Optical photographs taken after 24 hours of in vitro crosslinking of an IPTS (crosslinking agent-4) aqueous system;

[0041] Figure 6 The 10wt% PA-PEG-PA (coploymer-4) and 5wt% 4-arm-PEG disclosed in Example 39 of this invention 5000 - Frequency scan, strain scan, and time scan rheological curves of the IPTS (crosslinking agent-4) water system before and 24 hours after crosslinking. Detailed Implementation

[0042] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0043] The term "embodiment" used herein, as an example, is not necessarily to be construed as superior to or better than other embodiments. Performance testing in the embodiments of this application, unless otherwise specified, employs conventional testing methods in the art. It should be understood that the terminology used in this application is merely for describing particular implementations and is not intended to limit the scope of this disclosure.

[0044] Unless otherwise stated, the technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains; other experimental methods and technical means not specifically mentioned herein refer to experimental methods and technical means commonly used by one of ordinary skill in the art.

[0045] To better illustrate the content of this application, numerous specific details are provided in the following detailed embodiments. Those skilled in the art should understand that this application can be implemented even without certain specific details. In the embodiments, some methods, means, instruments, and devices well-known to those skilled in the art are not described in detail in order to highlight the main points of this application.

[0046] Without conflict, the technical features disclosed in the embodiments of this application can be combined arbitrarily, and the resulting technical solution belongs to the content disclosed in the embodiments of this application.

[0047] To better understand the present invention, the following embodiments are provided for further detailed description of the present invention, but they should not be construed as limiting the present invention. Any non-essential improvements and adjustments made by those skilled in the art based on the above-described invention are also considered to fall within the protection scope of the present invention.

[0048] Example 1

[0049] The synthesis steps of alkoxysilanized PEG crosslinking agent-1 are as follows: Take 5g of PEG 1000 In a three-necked flask, an appropriate amount of toluene was added for azeotropic dehydration. Heating was stopped once no bubbles appeared in the system. The system temperature was then maintained at 70°C under an inert gas atmosphere. 7.7 g of the reactant propyltrimethoxysilane isocyanate and 0.4 g of the catalyst stannous isooctanoate were added to the system, and the reaction was carried out under mechanical stirring for one hour. After the reaction was complete and the system cooled to room temperature, approximately 15 mL of dichloromethane was added to the system to fully dissolve the product. The solution was then slowly added dropwise to 1000 mL of ice-cold petroleum ether to allow precipitation. The precipitate was collected, washed three times with ice-cold petroleum ether, and dried under vacuum to obtain the alkoxysilane-modified PEG crosslinking agent-1.

[0050] Example 2

[0051] The synthesis steps of alkoxysilanized PEG crosslinking agent-2 are as follows: Take 20g of PEG 100000 In a three-necked flask, an appropriate amount of toluene was added for azeotropic dehydration. Heating was stopped once no bubbles appeared in the system. The system temperature was then maintained at 90°C under an inert gas atmosphere. 0.15 g of the reactant propyltriethoxysilane isocyanate and 0.1 g of the catalyst stannous isooctanoate were added to the system, and the reaction was carried out under mechanical stirring for one hour. After the reaction was complete and the system cooled to room temperature, approximately 15 mL of dichloromethane was added to the system to fully dissolve the product. The solution was then slowly added dropwise to 1000 mL of ice-cold petroleum ether to allow precipitation. The precipitate was collected and washed three times with ice-cold petroleum ether. After vacuum drying, the alkoxysilane-modified PEG crosslinking agent crosslinking agent-2 was obtained.

[0052] Example 3

[0053] The synthesis steps of alkoxysilanized PEG crosslinking agent-3 are as follows: Take 20g of PEG 5000 In a three-necked flask, an appropriate amount of toluene was added for azeotropic dehydration. Heating was stopped once no bubbles appeared in the system. The system temperature was then maintained at 70°C under an inert gas atmosphere. 2.5 g of the reactant propyltrimethoxysilane isocyanate and 0.2 g of the catalyst stannous isooctanoate were then added to the system, and the reaction was carried out under mechanical stirring for one hour. After the reaction was complete and the system cooled to room temperature, approximately 15 mL of dichloromethane was added to the system to fully dissolve the product. The solution was then slowly added dropwise to 1000 mL of ice-cold petroleum ether to allow precipitation. The precipitate was collected and washed three times with ice-cold petroleum ether. After vacuum drying, the alkoxysilane-modified PEG crosslinking agent crosslinking agent-3 was obtained.

[0054] Example 4

[0055] The synthesis steps of alkoxysilanized PEG crosslinking agent-4 are as follows: Take 20g of four-armed PEG 5000 In a three-necked flask, an appropriate amount of toluene was added for azeotropic dehydration. Heating was stopped once no bubbles appeared in the system. The system temperature was then maintained at 70°C under an inert gas atmosphere. 6g of the reactant propyltriethoxysilane isocyanate and 0.4g of the catalyst stannous isooctanoate were then added to the system, and the reaction was carried out under mechanical stirring for one hour. After the reaction was complete and the system cooled to room temperature, approximately 15mL of dichloromethane was added to the system to fully dissolve the product. The solution was then slowly added dropwise to 1000mL of ice-cold petroleum ether to allow precipitation. The precipitate was collected and washed three times with ice-cold petroleum ether. After vacuum drying, the alkoxysilane-modified PEG crosslinking agent crosslinking agent-4 was obtained.

[0056] Example 5

[0057] The synthesis steps of alkoxysilanized PEG crosslinking agent-5 are as follows: Take 20g of six-armed PEG 10000 In a three-necked flask, an appropriate amount of toluene was added for azeotropic dehydration. Heating was stopped once no bubbles appeared in the system. After cooling to room temperature under an inert gas atmosphere, approximately 50 mL of dichloromethane was added, followed by 4.5 g of the reactant propyltriphenoxysilane isocyanate and 0.4 g of the catalyst stannous isooctanoate. The mixture was then stirred at room temperature for 6 hours. After the reaction was complete, the solution was concentrated, precipitated with ice-cold petroleum ether, washed, and vacuum dried to obtain the alkoxysilanized PEG crosslinking agent crosslinking agent-5.

[0058] Example 6

[0059] The synthesis steps of alkoxysilanized PEG crosslinking agent-6 are as follows: Take 20g of eight-arm PEG 10000 In a three-necked flask, an appropriate amount of toluene was added for azeotropic dehydration. Heating was stopped once no bubbles appeared in the system. After cooling to room temperature under an inert gas atmosphere, approximately 50 mL of dichloromethane was added, followed by 6 g of the reactant propyltriacetoxysilane isocyanate and 0.4 g of the catalyst stannous isooctanoate. The mixture was then stirred at room temperature for 6 hours. After the reaction was complete, the solution was concentrated, precipitated with ice-cold petroleum ether, washed, and vacuum dried to obtain the alkoxysilanized PEG crosslinking agent crosslinking agent-6.

[0060] Referring to the preparation methods of Examples 1-6, the type of PEG, the terminal group of PEG, and the small molecules of the reaction were changed to obtain the corresponding alkoxysilanized PEG crosslinking agents. Their relevant molecular parameters are listed in Table 1 together with those of Examples 1-8.

[0061] Table 1:

[0062]

[0063]

[0064] Example 9

[0065] Amphiphilic polymer cploymer-1 (mPEG-PA) L The synthesis steps of mPEG are as follows: Take 1g of mPEG 2000 -NH2 and 0.13 g of 18-crown ether-6 were placed in a side-necked flask, and an appropriate amount of toluene was added for azeotropic dehydration. After the system cooled to room temperature, 50 mL of dichloromethane and 1.4 g of... L The alanine-NCA monomer was reacted at room temperature for 12 h under an argon atmosphere. After the reaction, the reaction solution was slowly added dropwise to 1000 mL of ice-cold diethyl ether to settle. The precipitate was collected by filtration and vacuum dried. The obtained solid was added to 200 mL of ultrapure water and stirred vigorously for 6 h. Then, it was centrifuged at 8000 r / min for 20 min, and the supernatant was lyophilized to obtain a powdered solid, Coplomer-1. The molecular weight and molecular weight distribution of the obtained product were characterized by 1H NMR spectroscopy and DMF phase gel permeation chromatography, and were found to be 3550 and 1.12, respectively.

[0066] Example 10

[0067] Amphiphilic polymer cploymer-2 (mPEG-PA) D The synthesis steps of mPEG are as follows: Take 1g of mPEG 2000-NH2 and 0.13 g of 18-crown ether-6 were placed in a side-necked flask, and an appropriate amount of toluene was added for azeotropic dehydration. After the system cooled to room temperature, 50 mL of dichloromethane and 1.4 g of... D The alanine-NCA monomer was reacted at room temperature for 12 h under an argon atmosphere. After the reaction, the reaction solution was slowly added dropwise to 1000 mL of ice-cold diethyl ether to settle. The precipitate was collected by filtration and vacuum dried. The obtained solid was added to 200 mL of ultrapure water and stirred vigorously for 6 h. Then, it was centrifuged at 8000 r / min for 20 min, and the supernatant was lyophilized to obtain the powdered solid Coplomer-2. The molecular weight and molecular weight distribution of the product were characterized by 1H NMR spectroscopy and DMF phase gel permeation chromatography, and were found to be 3560 and 1.13, respectively.

[0068] Example 11

[0069] Amphiphilic polymer cploymer-3 (PA) L -PEG-PA L The synthesis steps of NH2-PEG are as follows: Take 1g of NH2-PEG 2000 -NH2 and 0.26 g of 18-crown ether-6 were placed in a side-necked flask, and an appropriate amount of toluene was added for azeotropic dehydration. After the system cooled to room temperature, 50 mL of dichloromethane and 1.4 g of... L The alanine-NCA monomer was reacted at room temperature for 12 h under an argon atmosphere. After the reaction, the reaction solution was slowly added dropwise to 1000 mL of ice-cold diethyl ether to settle. The precipitate was collected by filtration and vacuum dried. The obtained solid was added to 200 mL of ultrapure water and stirred vigorously for 6 h. Then, it was centrifuged at 8000 r / min for 20 min, and the supernatant was lyophilized to obtain the powdered solid Coplomer-3. The molecular weight and molecular weight distribution of the product were characterized by 1H NMR spectroscopy and DMF phase gel permeation chromatography, which were 3360 and 1.13, respectively.

[0070] Example 12

[0071] Amphiphilic polymer cploymer-4 (PA) D -PEG-PA D The synthesis steps of NH2-PEG are as follows: Take 1g of NH2-PEG 2000 -NH2 and 0.26 g of 18-crown ether-6 were placed in a side-necked flask, and an appropriate amount of toluene was added for azeotropic dehydration. After the system cooled to room temperature, 50 mL of dichloromethane and 1.4 g of... DThe alanine-NCA monomer was reacted at room temperature for 12 h under an argon atmosphere. After the reaction, the reaction solution was slowly added dropwise to 1000 mL of ice-cold diethyl ether to settle. The precipitate was collected by filtration and vacuum dried. The obtained solid was added to 200 mL of ultrapure water and stirred vigorously for 6 h. Then, it was centrifuged at 8000 r / min for 20 min, and the supernatant was lyophilized to obtain the powdered solid Coplomer-4. The molecular weight and molecular weight distribution of the product were characterized by 1H NMR spectroscopy and DMF phase gel permeation chromatography, and were found to be 3500 and 1.13, respectively.

[0072] Example 13

[0073] Amphiphilic polymer cploymer-5 (PA) D -PEG-PA D The synthesis steps of NH2-PEG are as follows: Take 1g of NH2-PEG 5000 -NH2 and 0.11 g of 18-crown ether-6 were placed in a side-necked flask, and an appropriate amount of toluene was added for azeotropic dehydration. After the system cooled to room temperature, 50 mL of dichloromethane and 1.4 g of... D The alanine-NCA monomer was reacted at room temperature under an argon atmosphere for 12 h. After the reaction, the reaction solution was slowly added dropwise to 1000 mL of ice-cold diethyl ether to settle. The precipitate was collected by filtration and vacuum dried. The obtained solid was added to 200 mL of ultrapure water and stirred vigorously for 6 h. Then, it was centrifuged at 8000 r / min for 20 min, and the supernatant was lyophilized to obtain the powdered solid Coplomer-5. The molecular weight and molecular weight distribution of the product were characterized by 1H NMR spectroscopy and DMF phase gel permeation chromatography, which were 8560 and 1.11, respectively.

[0074] Example 14

[0075] Amphiphilic polymer cploymer-6 (mPEG-PA) L F L The synthesis steps of mPEG are as follows: Take 1g of mPEG 2000 -NH2 and 0.13 g of 18-crown ether-6 were placed in a side-necked flask, and an appropriate amount of toluene was added for azeotropic dehydration. After the system cooled to room temperature, 50 mL of dichloromethane and 1.1 g of... L -Alanine-NCA and 0.18g LThe phenylalanine-NCA monomer was reacted at room temperature for 12 h under an argon atmosphere. After the reaction, the reaction solution was slowly added dropwise to 1000 mL of ice-cold diethyl ether to settle. The precipitate was collected by filtration and vacuum dried. The obtained solid was added to 200 mL of ultrapure water and stirred vigorously for 6 h. Then, it was centrifuged at 8000 r / min for 20 min, and the supernatant was lyophilized to obtain the powdered solid Coplomer-6. The molecular weight and molecular weight distribution of the product were characterized by 1H NMR spectroscopy and DMF phase gel permeation chromatography, and were 3510 and 1.11, respectively.

[0076] Example 15

[0077] Amphiphilic polymer cploymer-7 (mPEG-PA) L E L The synthesis steps of mPEG are as follows: Take 1g of mPEG 2000 -NH2 and 0.13g of 18-crown ether-6 were placed in a side-necked flask, and an appropriate amount of toluene was added for azeotropic dehydration. After the system cooled to room temperature, 50mL of dichloromethane and 1.2g of... L -Alanine-NCA and 0.3 g of γ-benzyl-glutamate-NCA monomer were reacted at room temperature under an argon atmosphere for 12 h. After the reaction, the reaction solution was slowly added dropwise to 1000 mL of ice-cold diethyl ether to settle. The precipitate was collected by filtration and vacuum dried. The obtained solid was added to 200 mL of ultrapure water and stirred vigorously for 6 h. Then, it was centrifuged at 8000 r / min for 20 min, and the supernatant was lyophilized to obtain powdered solid Coplomer-7. The molecular weight and molecular weight distribution of the product were characterized by 1H NMR spectroscopy and DMF phase gel permeation chromatography, which were 3710 and 1.12, respectively.

[0078] Example 16

[0079] Amphiphilic polymer cploymer-8 (mPEG-PA) D D D The synthesis steps of mPEG are as follows: Take 1g of mPEG 4000 -NH2 and 0.07g of 18-crown ether-6 were placed in a side-necked flask, and an appropriate amount of toluene was added for azeotropic dehydration. After the system cooled to room temperature, 50mL of dichloromethane and 1.2g of... D -Alanine-NCA and 0.3g DThe γ-benzyl-aspartic acid ester-NCA monomer was reacted at room temperature under an argon atmosphere for 12 h. After the reaction, the reaction solution was slowly added dropwise to 1000 mL of ice-cold diethyl ether to settle. The precipitate was collected by filtration and vacuum dried. The obtained solid was added to 200 mL of ultrapure water and stirred vigorously for 6 h. Then, it was centrifuged at 8000 r / min for 20 min, and the supernatant was lyophilized to obtain the powdered solid Coplomer-8. The molecular weight and molecular weight distribution of the product were characterized by 1H NMR spectroscopy and DMF phase gel permeation chromatography, which were 3280 and 1.07, respectively.

[0080] Example 17

[0081] Amphiphilic polymer cploymer-9 (mPEG-PMeD) D BnD D The synthesis steps of mPEG are as follows: Take 1g of mPEG 2000 -NH2 and 0.13 g of 18-crown ether-6 were placed in a side-necked flask, and an appropriate amount of toluene was added for azeotropic dehydration. After the system cooled to room temperature, 50 mL of dichloromethane and 0.7 g of... D -β-methyl-aspartic acid ester-NCA monomer and 0.7g D -β-ethyl-aspartic acid ester-NCA was reacted at room temperature under an argon atmosphere for 12 h. After the reaction was completed, the reaction solution was slowly added dropwise to 1000 mL of ice-cold diethyl ether to settle. The precipitate was collected by filtration and vacuum dried. The obtained solid was added to 200 mL of ultrapure water and stirred vigorously for 6 h. Then, it was centrifuged at 8000 r / min for 20 min, and the supernatant was lyophilized to obtain the powdered solid Coplomer-9. The molecular weight and molecular weight distribution of the product were characterized by 1H NMR spectroscopy and DMF phase gel permeation chromatography, which were 4700 and 1.18, respectively.

[0082] Examples 18-21

[0083] Following the preparation methods of Examples 9-17, the molecular weight of the hydrophilic blocks and the types and proportions of the hydrophobic blocks were varied to obtain corresponding amphiphilic block polymers. Their relevant molecular parameters are listed in Table 2, along with those of Examples 9-17.

[0084] Table 2:

[0085]

[0086]

[0087] Example 22

[0088] Coplymer-10.6g was fully dissolved in a certain mass of pure water. Then, 1g of alkoxysilane-modified PEG crosslinking agent crosslinking agent-1 and 1g of functional additive iohexol were added to the system. The total mass of the system was adjusted to 10g by adjusting the mass of the solvent, and the pH of the system was adjusted to 7. The system was placed in a 4℃ refrigerator and stirred thoroughly. Then, it was placed in a 37℃ air bath for crosslinking for 100h to obtain Dual crosslinked hydrogel-1.

[0089] Example 23

[0090] Coplymer-2 1.3g was fully dissolved in a certain mass of physiological saline. Then, 0.5g of alkoxysilanized PEG crosslinking agent crosslinking agent-2 and 3g of functional additive iodine oil were added to the system. The total mass of the system was adjusted to 10g by adjusting the mass of the solvent. The system was placed in a 4℃ refrigerator and stirred thoroughly. After that, it was placed in a 37℃ air bath for 1 hour to crosslink, thus obtaining Dual crosslinked hydrogel-2.

[0091] Example 24

[0092] Coplymer-31g was fully dissolved in a certain mass of phosphate buffer solution. Then, 0.5g of alkoxysilanized PEG crosslinking agent crosslinking agent-3, 3g of functional auxiliary agent iopromide, chitin regulator, and 0.75g each of xylitol were added to the system. The total mass of the system was adjusted to 10g by adjusting the mass of the solvent, and the pH value of the system was adjusted to 10. The system was placed in a 4℃ refrigerator and stirred thoroughly. Then, it was placed in a 37℃ air bath for crosslinking for 4h to obtain Dualcrosslinked hydrogel-3.

[0093] Examples 25-34

[0094] Following the preparation methods of Examples 22-24, the composition and proportions of the synthesized amphiphilic polymer, alkoxysilanized PEG crosslinking agent, functional additives, regulators, and water-based solvents were changed to obtain corresponding bicrosslinked hydrogels. The relevant components and proportions are listed in Table 3, along with those in Examples 22-24.

[0095] Table 3:

[0096]

[0097]

[0098]

[0099] Example 35

[0100] Dual crosslinked hydrogel-1 and Dual crosslinked hydrogel-2 contain iodine-containing iohexol and iodized oil, thus possessing X-ray imaging capabilities and can be used for vascular embolization.

[0101] Example 36

[0102] The synthesized triethoxysilanized four-arm PEG 5000 (Crosslinking agent-4) was prepared as a 5wt% PBS solution, and the pH of the solution was adjusted to 8, 9, 10, and 11. The solutions were incubated in a 37°C water bath with a shaker. Crosslinking was determined by observation: the transparent glass bottle containing the sample was inverted 180°; if the sample flowed down, crosslinking had not occurred; otherwise, crosslinking was considered to have occurred. At pH 8, crosslinking took approximately 76 hours. Increasing the pH accelerated the crosslinking rate. At pH 9 and 10, crosslinking took 19 and 4 hours, respectively. Temperature also affects the crosslinking rate of alkoxysilanized PEG. A 5wt% PBS solution of crosslinking agent-4, adjusted to pH 10, was incubated at 4, 25, 30, and 37°C. As the temperature increased, the required crosslinking time gradually decreased. The experimental results are shown below. Figure 3 As shown in the figure. Based on this result, it is demonstrated that the rate of the chemical cross-linking reaction can be effectively controlled by adjusting the temperature and pH value of the entire system. Therefore, the chemical cross-linking time can vary from a few minutes to several days depending on the application scenario.

[0103] Example 37

[0104] Configured with 10wt% PA-PEG-PA (coploymer-4) and 5wt% 4-arm-PEG 5000 The shear-thinning properties of the PBS water system containing IPTS (crosslinking agent-4) were tested using a rotational rheometer with a cone plate of 60 mm diameter, 1° angle, and 0.03 mm gap height. The shear rates were measured in the range of 0.1–1000 s⁻¹ at 37 °C, an oscillation frequency of 1.592 Hz, and a strain of 1%. -1 The shear viscosity of the aqueous system decreased by approximately three orders of magnitude with increasing shear rate. Furthermore, the aqueous system after 4 hours of crosslinking at room temperature also exhibited shear thinning, with a decrease in shear viscosity of approximately four orders of magnitude. The data are as follows: Figure 4 As shown, this demonstrates that the water system proposed in this patent has shear-thinning ability before complete chemical cross-linking, thus allowing for better delivery to the designated site via needles or catheters.

[0105] Example 38

[0106] Configured with 10wt% PA-PEG-PA (coploymer-4) and 5wt% 4-arm-PEG 5000 A PBS-water system containing IPTS (crosslinking agent-4) was carefully added dropwise to a glass sample vial. The vial was then placed in a 37°C water bath shaker for 24 hours to fully crosslink. Afterward, the glass sample vial was broken to remove the crosslinked hydrogel, which was then photographed. The crosslinked hydrogel exhibited a complete and stable shape, as shown in the image. Figure 5 As shown.

[0107] Example 39

[0108] Configured with 10wt% PA-PEG-PA (coploymer-4) and 5wt% 4-arm-PEG 5000 The rheological properties of hydrogels in a PBS-water system containing IPTS (crosslinking agent-4) before and after crosslinking at 37℃ for 24 hours in vitro were compared. An 8mm diameter plate was used as the clamp. Time scans were performed at 1.592Hz with a strain of 1%, frequency scans at 0.01–10Hz with a strain of 1%, and strain scans at 0.01–100% with a frequency of 1.592Hz. The test temperature was 37℃. The G′ (elastic modulus) of the crosslinked hydrogel sample was significantly higher than that before crosslinking, and the elastic modulus remained very stable during the 10-minute temperature scan. This is attributed to the reinforcing effect of the chemical crosslinking network on the hydrogel. The results are as follows: Figure 6 As shown.

[0109] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A physically / chemically cross-linked hydrogel, characterized in that, The raw materials include the following parts by weight: 6-13 parts of amphiphilic polymer, 1-10 parts of alkoxysilane-modified PEG crosslinking agent, 0-30 parts of functional additives, and 47-93 parts of water-based solvent; the amphiphilic polymer uses polyethylene glycol or polyvinylpyrrolidone as hydrophilic blocks and polyamino acids as hydrophobic blocks; the structure of the alkoxysilane-modified PEG crosslinking agent is that the PEG polymer is modified with alkoxysilane crosslinking groups at the ends.

2. The physical / chemical dual crosslinked hydrogel according to claim 1, characterized in that, The molecular weight of the PEG polymer ranges from 400 to 100,000, including one or more of linear PEG, 4-arm PEG, 6-arm PEG, and 8-arm PEG; the alkoxysilyl crosslinking groups modified at the ends of the polymer include one or more of trimethoxysilyl, dimethoxymethylsilyl, triethoxysilyl, diethoxymethylsilyl, triphenoxysilyl, diphenoxymethylsilyl, triacetoxysilyl, and diacetoxymethylsilyl.

3. The physical / chemical double crosslinked hydrogel according to claim 1, characterized in that, The amphiphilic polymer comprises 40 wt% to 60 wt% of hydrophilic blocks and 60 wt% to 40 wt% of hydrophobic block polyamino acids.

4. The physical / chemical dual crosslinked hydrogel according to claim 3, characterized in that, The hydrophilic block is polyethylene glycol with a molecular weight range of 750-5000 or polyvinylpyrrolidone with a molecular weight range of 750-5000; the hydrophobic block is polyamino acid with a molecular weight range of 400-5000.

5. The physical / chemical dual crosslinked hydrogel according to claim 4, characterized in that, The polyamino acid includes L or D -Alanine, L or D -Phenylalanine, L or D -Leucine, L or D -Isoleucine, L or D -valine, L or D -Lysine, L or D -Aspartic acid, L or D -Aspartic acid methyl ester L or D - Ethyl aspartate L or D -Aspartic acid benzyl ester L or D -Glutamic acid, L or D - glutamate methyl ester L or D -Ethyl glutamate, L or D Homopolymers of β-benzyl glutamate, or the above L Type or D A copolymer formed from monomers in any form.

6. The physical / chemical dual crosslinked hydrogel according to claim 3, characterized in that, The amphiphilic polymer is selected from one or more of the following: AB diblock copolymer, BAB triblock copolymer, AgB-type grafted block copolymer, and n-arm-(AB) type N-arm block copolymer, wherein n is an integer greater than 2 and less than 10.

7. The physical / chemical dual crosslinked hydrogel according to claim 1, characterized in that, The functional additives include: X-ray contrast agents: one or more combinations of iodized oil, octyl triiodobenzoate, iohexol, iopamidol, iodixanol, iofluoxetine, iodamide, and iopromide; or, Fluorescent dyes: one or more combinations of Rhodamine, Nile Red, Cy3, Cy5, Cy5.5, Cy7, and tetraphenylethylene; or, Magnetic resonance imaging contrast agents: gadopentetate dimeglumine, gadofusamide, gadobutrol, superparamagnetic iron oxide particles; or, Anticancer drugs: one or more of the following: doxorubicin, paclitaxel, docetaxel, vinorelbine, gemcitabine, 5-fluorouracil, capecitabine, sorafenib, lenvatinib, ramucirumab, apatinib, cisplatin, oxaliplatin, bleomycin, regorafenib, cabozantinib, irinotecan, and nivolumab; or, Diabetes medications: one or more of the following: insulin, metformin, liximab, liraglutide, and exenatide; or, Antibacterial drugs: one or more of penicillin and gentamicin, or a combination thereof.

8. The physical / chemical dual crosslinked hydrogel according to claim 1, characterized in that, The water-based solvent is one or more of the following: pure water, physiological saline, phosphate buffer solution, cell culture medium, and tissue culture medium.

9. The application of the physical / chemical double crosslinked hydrogel as described in claim 1 in the preparation of materials for vascular embolization, tissue repair, cell encapsulation, and wound healing.

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