CN113929792B - Aldehyde modified hyaluronic acid (sodium) and synthesis method and application thereof - Google Patents

Aldehyde modified hyaluronic acid (sodium) and synthesis method and application thereof Download PDF

Info

Publication number
CN113929792B
CN113929792B CN202010671220.2A CN202010671220A CN113929792B CN 113929792 B CN113929792 B CN 113929792B CN 202010671220 A CN202010671220 A CN 202010671220A CN 113929792 B CN113929792 B CN 113929792B
Authority
CN
China
Prior art keywords
hydrazide
hyaluronic acid
modified
group
sodium hyaluronate
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN202010671220.2A
Other languages
Chinese (zh)
Other versions
CN113929792A (en
Inventor
阿斯根
徐倩
薛秀生
王文新
徐勇猛
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Fu Langfu Hangzhou Biological Technology Co ltd
Original Assignee
Fu Langfu Hangzhou Biological Technology Co ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Fu Langfu Hangzhou Biological Technology Co ltd filed Critical Fu Langfu Hangzhou Biological Technology Co ltd
Priority to CN202010671220.2A priority Critical patent/CN113929792B/en
Publication of CN113929792A publication Critical patent/CN113929792A/en
Application granted granted Critical
Publication of CN113929792B publication Critical patent/CN113929792B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
    • C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
    • C08B37/0027—2-Acetamido-2-deoxy-beta-glucans; Derivatives thereof
    • C08B37/003—Chitin, i.e. 2-acetamido-2-deoxy-(beta-1,4)-D-glucan or N-acetyl-beta-1,4-D-glucosamine; Chitosan, i.e. deacetylated product of chitin or (beta-1,4)-D-glucosamine; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
    • A61K8/73—Polysaccharides
    • A61K8/735—Mucopolysaccharides, e.g. hyaluronic acid; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
    • A61L26/0023—Polysaccharides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0061—Use of materials characterised by their function or physical properties
    • A61L26/008—Hydrogels or hydrocolloids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
    • C08J3/03—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media
    • C08J3/075—Macromolecular gels
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/91—Injection
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2305/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
    • C08J2305/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof

Abstract

The invention relates to novel aldehyde modified hyaluronic acid (sodium) and a preparation method and application thereof. The hyaluronic acid (sodium) modified by aldehyde group is prepared by performing aldehyde group functionalization on carboxyl of hyaluronic acid (sodium).

Description

Aldehyde modified hyaluronic acid (sodium) and synthesis method and application thereof
Technical Field
The invention belongs to the fields of biomedicine and medical cosmetology, and particularly relates to novel aldehyde modified hyaluronic acid (sodium) and a preparation method and application thereof.
Background
Hyaluronic acid, hydraronic acid, abbreviated as HA, is a high molecular weight biopolysaccharide that was first isolated in 1934 by professor kari meier and his assistant john paler in the vitreous of bovine eyes. Hyaluronic acid is a naturally occurring biopolymer with important biological functions in bacteria and higher animals including humans. HA is in most cases present in connective tissue, and is present in high amounts in particular in synovial fluid, vitreous humor of the eyeball, umbilical cord and cockscomb. HA is synthesized by a class of intact membrane proteins called hyaluronidase enzymes, and can be degraded by a range of hyaluronidase enzymes. Hyaluronic acid is available in two ways: one approach is through animal tissue extraction, such as: the hyaluronic acid extracted from the animal tissues of bulls eyes, cockscombs and the like often contains exogenous proteins, so the purity of the hyaluronic acid cannot meet the application in the biomedical field. Therefore, another method for preparing hyaluronic acid using microbial fermentation has been developed to solve the problem of purity.
HA HAs many applications in clinical, tissue engineering repair, and gene and drug delivery. Hyaluronic acid HAs good biocompatibility, so that the hyaluronic acid plays an important role in biological development and body injury repair processes, but HA HAs the defects of excessively strong water solubility, short metabolic cycle and the like in a human body, and can be crosslinked to overcome the defects. The HA is crosslinked, so that the in vivo maintenance time of the HA can be greatly prolonged, and the action effect can be prolonged. Commercial crosslinked HA products are mainly prepared by reacting two small molecule crosslinkers, butanediol diglycidyl ether and divinyl sulfone, with hydroxyl groups in HA. Both crosslinking agents can cause denaturation of proteins in the human body, thereby causing allergic reactions. Therefore, the development of a novel hyaluronic acid crosslinking technology, and the development of a novel hyaluronic acid derivative and a crosslinked product thereof have wide application prospects and commercial values.
In the prior art, the hydrogel is generally prepared by crosslinking hyaluronic acid with a small-molecule crosslinking agent, and after the crosslinking reaction is completed in vitro and the small-molecule crosslinking agent is purified and removed, the hydrogel can be applied to organisms, so that the hydrogel has obvious disadvantages including: 1. hydrogels are too viscous to or difficult to inject through standard bore injection needles, and the replacement of large bore injection needles can increase patient pain and increase the risk of inflammation and trauma. 2. The physical and chemical properties of the hydrogel are relatively fixed and cannot be well matched with the specific requirements in the fields of biological medicine and medical cosmetology. 3. The small-molecule cross-linking agent often remains in the hydrogel, because part of the small-molecule cross-linking agent is locked in the three-dimensional space structure of the hydrogel in the preparation process and cannot be completely removed in the purification process of the hydrogel, and the residues of the small-molecule cross-linking agent such as butanediol diglycidyl ether and divinyl sulfone bring safety risks and adverse reaction events, thereby restricting the application of the hydrogel in the fields of biomedicine and medical cosmetology.
Researchers in the related art have made many efforts to develop in situ cross-linked hydrogel compositions suitable for various application needs, which are injected into organisms in a liquid form rather than in a hydrogel form, and the cross-linking reaction thereof to form the hydrogel is completed in the organisms. Because it is in a flowable liquid form prior to injection, when injected into a target site in a living being, a hydrogel is formed that conforms substantially to the shape of the target site. The injectable property not only makes the operation process simple and convenient, but also can avoid the pain of the patient caused by the implantation operation, and greatly reduce the traumatic property of the operation. For example, WO 95/15168 reports the synthesis of HA hydrazide derivatives by chemical coupling using 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide, EDC, and hydrazide-HA can be crosslinked using homo-or hetero-bifunctional or Traut's crosslinkers to form hydrogels for drug delivery. WO 00/016818 discloses in situ formation of hydrogels by crosslinking aldehyde or amine functionalized derivatives of HA (e.g., adipic dihydrazide-HA) with homo-or hetero-bifunctional crosslinkers (e.g., bifunctional N-hydroxysuccinimide ester crosslinkers such as (SPA) 1-PEG). Dahlmann et al (Biomaterials, 2013,34, 940-951) describe in situ crosslinkable alginate and HA hydrogels for myocardial tissue engineering prepared by reacting aldehyde and hydrazide functionalized alginate and HA in the presence of human class I collagen and neonatal rat cardiac cells (NRHC) to produce hydrazone crosslinked hydrogel-based bioartificial heart tissue.
The aldehyde group functionalized hyaluronic acid is a hyaluronic acid derivative which is easy to crosslink, good in biocompatibility, good in stability and easy to store. In recent years, various documents report that the hydrogel formed by the hyaluronic acid derivative shows good application value in the fields of tissue engineering and regenerative medicine. However, the synthesis method of aldehyde modified hyaluronic acid reported at present is mainly obtained by oxidizing vicinal diol sites in HA disaccharide unit. This method inevitably hydrolyzes the glycosidic bond of hyaluronic acid, thereby causing a decrease in the molecular weight of hyaluronic acid, which in turn leads to a decrease in the mechanical properties of the end product. This problem greatly hinders the progress of the aldehyde-based hyaluronic acid derivatives in industrial applications.
Disclosure of Invention
The invention provides a method for preparing aldehyde modified hyaluronic acid (sodium) by aldehyde functionalization of carboxyl of hyaluronic acid (sodium), the aldehyde modified hyaluronic acid (sodium) method for carboxyl of hyaluronic acid (sodium) can not cause hydrolysis of glycosidic bond of hyaluronic acid (sodium), the reaction is mild and controllable, high temperature is not needed, and byproducts are easy to remove.
The aldehyde modified hyaluronic acid (sodium) can form hydrogel after crosslinking, and particularly, the hydrogel forming mode can be realized in a self-crosslinking mode without using a small-molecule crosslinking agent, so that in-situ crosslinking in a living body can be realized, various defects of the hydrogel using the small-molecule crosslinking agent (butanediol diglycidyl ether, divinyl sulfone and the like) in the prior art are avoided, and a better choice is provided for the application of the hydrogel in the fields of biomedicine and medical cosmetology.
In addition, the aldehyde hyaluronic acid (sodium) can also be subjected to Schiff base condensation reaction with a micromolecule cross-linking agent containing amino or hydrazide groups to form hydrogel, the cross-linking reaction is dynamic and reversible, and the obtained hydrogel has the characteristics of self-healing and shear thinning: the self-healing has important significance in solving the damage repair problem of the flexible biological material and realizing the intelligent and efficient development of the biological material; the shear thinning properties give the hydrogel good injectability. During colloid injection, the gel is changed into a solution form by the tangential shearing force, and the solution form is restored to the gel form when the external shearing force disappears after the colloid passes through the needle tube. The performance can well prevent cells in the colloid from being damaged by high shearing force. The self-healing and shear thinning lead the hydrogel to have attractive application prospect in the field of injection filling molding cosmetic products.
According to one aspect of the present invention, the present invention relates to an aldehyde modified hyaluronic acid or sodium hyaluronate having the structure represented by the following formula:
Figure BDA0002582369280000041
it will be appreciated by those skilled in the art that hyaluronic acid or sodium hyaluronate of various molecular weights may be used in the present invention. In a preferred embodiment, the hyaluronic acid or sodium hyaluronate modified by hydroformylation has a molecular weight of 10-2200kDa, preferably 50-1000kDa, further preferably 90-110kDa, most preferably about 100kDa.
In a preferred embodiment, the modification rate (or degree of substitution) of the aldehyde-modified hyaluronic acid or sodium hyaluronate is 10-90%, preferably 50-80%, most preferably 60-70%.
It will be appreciated by those skilled in the art that hyaluronic acid (sodium) from a variety of sources may be used in the present invention, either commercially available or self-made, as an extract, or prepared by biological fermentation or the like.
According to another aspect of the invention, the invention relates to a preparation method of the aldehyde modified hyaluronic acid or sodium hyaluronate, which comprises the following steps:
(1) Dissolving hyaluronic acid (sodium) in water and adjusting the pH of the resulting solution to 4.5-5.0, preferably 4.7-4.8, adding EDCI (1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride) and stirring to dissolve it, followed by adding the propylene glycol hydrazide derivative 2,3-dihydroxypropyl-3- ((3-hydrazino-3-oxopropyl) thio) -2-methylpropionate (2, 3-dihydroxypropyl-3- ((3-hydrozinyl-3-oxopropyl) thio) -2-methylpropionate), reacting at 0 ℃ -90 ℃, preferably 20 ℃ -50 ℃, most preferably stirring at room temperature for 0.5-12h, preferably 3-5h, most preferably about 4h, adjusting the pH of the reaction solution to 6.0-8.0, preferably 6.9-7.1 to terminate the reaction, dialyzing the reaction solution and lyophilizing to obtain propylene glycol modified hyaluronic acid (sodium);
preferably, the pH of the reaction solution is controlled to be 4.5-5.0, more preferably 4.7-4.8 during the reaction;
preferably, the molecular weight of the hyaluronic acid (sodium) is between 10 and 2200kDa, preferably between 50 and 1000kDa, further preferably between 90 and 110kDa, most preferably about 100kDa;
preferably, the mass/volume concentration of the aqueous hyaluronic acid (sodium) solution is between 0.1% and 2.5%, more preferably between 0.5 and 1.5%, most preferably about 1.0%;
preferably, the mass ratio of EDCI to hyaluronic acid (sodium) is 1;
preferably, the EDCI is added to dissolve followed by 2 to 60 minutes of continued stirring, more preferably 5 to 30 minutes, most preferably about 15 minutes before the addition of the propylene glycol hydrazide derivative to activate the reactive sites to be modified;
preferably, the mass ratio of the propylene glycol hydrazide derivative to hyaluronic acid (sodium) is 1;
as will be appreciated by those skilled in the art, dialysis is used to remove reaction residues and impurities. Preferably, a dialysis bag is used.
Preferably, lyophilization is carried out until the water content is below 10%.
(2) Dissolving the propylene glycol modified hyaluronic acid (sodium) obtained in the step (1) in water, and adding sodium periodate (NaIO) 4 ) Sodium silicate (Na) 2 SiO 3 ) Aqueous solution, protected from light at 0-90 deg.C, preferably 20-50 deg.C, most preferably room temperature under stirring for 2-72h, preferably 16-32h, most preferablySelecting about 24h, adding ethylene glycol, stirring for 0.2h-4h, preferably 0.5-2h, most preferably about 1h to terminate the reaction, dialyzing the reaction solution, and freeze-drying to obtain aldehyde modified hyaluronic acid (sodium).
Preferably, the mass/volume concentration of the aqueous solution of propylene glycol modified hyaluronic acid (sodium) is from 0.1% to 2.5%, more preferably from 0.5 to 1.5%, most preferably about 1.0%;
preferably, the mass ratio of sodium periodate to propylene glycol modified hyaluronic acid (sodium) is 1;
preferably, sodium periodate is sodium silicate (Na) 2 SiO 3 ) A mass/volume concentration in the aqueous solution of 0.5% to 15%, more preferably 8 to 12%, most preferably about 10%;
preferably sodium silicate (Na) 2 SiO 3 ) The mass/volume concentration of sodium silicate in the aqueous solution is 0.2% to 5%, more preferably 0.5 to 1.5%, most preferably about 1.0%;
one skilled in the art will appreciate that dialysis is performed to remove reaction residues and impurities. Preferably, a dialysis bag is used.
Preferably, lyophilization is carried out until the water content is below 10%.
It will be appreciated by those skilled in the art that various pH adjusting agents commonly used in the art may be used in the present invention. Preferably, the pH is adjusted using an aqueous hydrochloric acid solution and/or an aqueous sodium hydroxide solution, more preferably, the concentration of the aqueous hydrochloric acid solution is about 1M and the concentration of the aqueous sodium hydroxide solution is about 1M.
The specific reaction route of the above reaction is as follows:
Figure BDA0002582369280000071
the aldehyde modified hyaluronic acid or sodium hyaluronate can perform self-crosslinking reaction with a polymer containing amino or hydrazide groups without a micromolecular crosslinking agent, so that in-situ crosslinking in a living body can be realized.
According to another aspect of the invention, the invention relates to a composition comprising an aldehydic modified hyaluronic acid or sodium hyaluronate according to the invention and an amino or hydrazide group-containing polymer. Preferably, the composition further comprises water. Preferably, the composition is free of cross-linking agents, including small molecule cross-linking agents. Preferably, the composition consists of the aldehydized modified hyaluronic acid or sodium hyaluronate of the present invention and a polymer containing an amino group or a hydrazide group. Preferably, the composition consists of the aldehydized modified hyaluronic acid or sodium hyaluronate of the present invention, an amino group-or hydrazide group-containing polymer and water.
According to another aspect of the present invention, the present invention relates to a kit comprising an aldehydized modified hyaluronic acid or sodium hyaluronate and an amino or hydrazide group-containing polymer, which are present independently of each other. Preferably, the kit further comprises water independently present. Preferably, the kit does not contain a crosslinking agent, including a small molecule crosslinking agent.
According to the present invention, in the above-mentioned composition and kit, an amino group-containing polymer such as chitosan, collagen, gelatin, etc. One skilled in the art can select the appropriate molecular weight of these amino group-containing polymers as desired. For example, the molecular weight is 10-2200kDa, preferably 50-1000kDa, more preferably 90-110kDa, and most preferably about 100kDa.
According to the present invention, in the above-mentioned composition and kit, the hydrazide group-containing polymer such as hydrazide-modified biocompatible polymer compound includes, but is not limited to, hydrazide-modified hyaluronic acid or sodium hyaluronate, hydrazide-modified chondroitin sulfate, hydrazide-modified heparin, hydrazide-modified sodium alginate, hydrazide-modified cellulose, hydrazide-modified gelatin, hydrazide-modified polyethylene glycol, hydrazide-modified dendrimer polyamide, and the like. One skilled in the art can select the appropriate molecular weight of these hydrazide group-containing polymers as desired. For example, the molecular weight is 10-2200kDa, preferably 50-1000kDa, more preferably 90-110kDa, and most preferably about 100kDa.
According to the present invention, in the above composition and kit, the mass ratio of the aldehyde modified hyaluronic acid or sodium hyaluronate to the amino or hydrazide group-containing polymer is 1.
According to the present invention, in the above composition, the mass/volume concentration of the aldehyde-modified hyaluronic acid or sodium hyaluronate is 0.1% to 2.5%, more preferably 0.5% to 1.5%, most preferably about 1.0%, and the mass/volume concentration of the amino or hydrazide group-containing polymer is 0.1% to 2.5%, more preferably 0.5% to 1.5%, most preferably about 1.0%.
It will be understood by those skilled in the art that the above compositions may be formulated prior to use, or that the components of the above kits, which are separate from each other, may be mixed and formulated into a composition prior to use. In general, the crosslinking reaction occurs immediately after the composition is formulated and is completed in a time period of about 2 minutes to 2 hours. It will be appreciated by those skilled in the art that the above composition may be injected into an organism at any time after the formulation is complete until the crosslinking reaction is complete.
According to another aspect of the present invention, the present invention relates to a hydrogel obtained by crosslinking the aldehyde-modified hyaluronic acid or sodium hyaluronate with the amino group-or hydrazide group-containing polymer.
According to the invention, the mass ratio of the aldehyde modified hyaluronic acid or sodium hyaluronate to the amino or hydrazide group-containing polymer is 1.
According to another aspect of the present invention, the present invention relates to a method for preparing the above hydrogel, comprising the steps of: preparing an aqueous solution containing the aldehyde modified hyaluronic acid or sodium hyaluronate and the amino or hydrazide group-containing polymer, and reacting at 0 ℃ to 90 ℃, preferably 20 ℃ to 50 ℃, and most preferably at room temperature.
According to the present invention, the mass/volume concentration of the formylation-modified hyaluronic acid or sodium hyaluronate in solution is 0.1% to 2.5%, more preferably 0.5% to 1.5%, most preferably about 1.0%, and the mass/volume concentration of the amino or hydrazide group-containing polymer is 0.1% to 2.5%, more preferably 0.5% to 1.5%, most preferably about 1.0%.
The aldehyde modified hyaluronic acid or sodium hyaluronate can be crosslinked under the action of a small molecule crosslinking agent, or crosslinked with an acrylate modified biocompatible high molecular compound.
According to another aspect of the invention, the invention relates to a hydrogel, which is obtained by crosslinking the aldehyde modified hyaluronic acid or sodium hyaluronate and a small molecule crosslinking agent containing amino or hydrazide groups.
According to another aspect of the invention, the invention relates to a hydrogel, which is obtained by crosslinking the aldehyde modified hyaluronic acid or sodium hyaluronate, the acrylate modified biocompatible macromolecular compound and the amino or hydrazide group-containing small molecule crosslinking agent.
According to the present invention, the amino group-containing small molecule crosslinking agent is, for example, a small molecule compound containing two or more primary amines or a physiologically acceptable salt thereof, a small molecule compound containing two or more primary amines such as cystine, lysine, ethylenediamine, propylenediamine, butylenediamine, pentylenediamine, hexylenediamine, p-phenylenediamine, m-phenylenediamine, p-phenylenediamine, biphenyldiamine, etc., a salt such as hydrochloride, sulfate, methanesulfonate, citrate, etc.
According to the present invention, the small molecule crosslinking agent containing a hydrazide group is, for example, a small molecule compound containing two or more hydrazides, such as oxalic acid dihydrazide, malonic acid dihydrazide, succinic acid dihydrazide, glutaric acid dihydrazide, adipic acid dihydrazide, phthalic acid dihydrazide, isophthalic acid dihydrazide, terephthalic acid dihydrazide, or the like.
According to another aspect of the present invention, the present invention relates to a method for preparing the above hydrogel, comprising the steps of: preparing an aqueous solution containing the aldehyde modified hyaluronic acid or sodium hyaluronate and an aqueous solution containing the amino or hydrazide group-containing small molecule cross-linking agent, dripping the aqueous solution containing the amino or hydrazide group-containing small molecule cross-linking agent into the aqueous solution containing the aldehyde modified hyaluronic acid or sodium hyaluronate, and stirring at 0-90 ℃, preferably 20-50 ℃ and most preferably at room temperature until the cross-linking reaction is completed.
Preferably, the aqueous solution containing the aldehyde-modified hyaluronic acid or sodium hyaluronate further contains an acrylate-modified biocompatible polymer compound.
The hydrogel can be applied to the fields of biomedicine and medical cosmetology, for example, the hydrogel can be prepared into external wound dressings, nose and lip filling for medical cosmetology, skeleton materials for replacing hyaluronic acid injection in the existing medical cosmetology and moderate-to-deep venous ulcer wound dressings, and the like.
The invention has the beneficial effects that:
1. the aldehyde functional group is introduced into the carboxyl position of hyaluronic acid (sodium) creatively;
2. the irreversible degradation reaction caused by introducing aldehyde group into the adjacent diol site of hyaluronic acid (sodium) in the prior art is avoided;
3. the first step of modification reaction has mild conditions and high modification efficiency;
4. the degree of the second oxidation reaction step is quantitative and controllable, and toxic byproducts are not generated;
5. the use of organic solvent is avoided, the residue problem of the organic solvent in the product is fundamentally eliminated, the biocompatibility is better, and the occurrence probability of rejection reaction and inflammatory reaction of implanted organisms such as human bodies can be greatly reduced.
Drawings
FIG. 1 is an infrared spectrum of the hydrazide-modified sodium hyaluronate prepared in example 1, 1717cm -1 The peak is an aldehyde group.
Detailed Description
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Furthermore, it should be understood that various changes or modifications can be made by those skilled in the art after reading the description of the present invention, and such equivalents also fall within the scope of the invention.
Preparation example 1 preparation of sodium hyaluronate hydrazoylated and modified
Dissolving 1.0g of sodium hyaluronate with the molecular weight of 100kDa in 100mL of purified water to obtain 1.0% sodium hyaluronate aqueous solution, adding 18.3g of adipic dihydrazide, and adjusting the pH value of the solution to 4.75 by using 1M HCl aqueous solution; adding 1.7g of EDCI, stirring and reacting for 2 hours at room temperature, adjusting the pH of the reaction solution to 7.0 by using 1M NaOH aqueous solution to stop the reaction, dialyzing and purifying the reaction solution, and freeze-drying to obtain the sodium hyaluronate modified by hydrazoylation, wherein the hydrazoylation modification rate is 78% by measurement.
Example 1 two-step Synthesis of sodium hyaluronate modified by hydroformylation
(1) Modification reaction: adding 50.0g of sodium hyaluronate with the molecular weight of 100kDa into 5000mL of ultrapure water, magnetically stirring to dissolve the sodium hyaluronate to obtain 1% (w/v) of clear transparent solution, and adjusting the pH of the clear transparent solution to 4.75 by using a proper amount of 1M HCl aqueous solution; adding 100.0g of EDCI, magnetically stirring to dissolve the EDCI, and continuously stirring for 15 minutes; adding 200.0g of propylene glycol hydrazide derivative 2,3-dihydroxypropyl-3- ((3-hydrazino-3-oxopropyl) thio) -2-methylpropionate, magnetically stirring at room temperature for 4 hours, and controlling the pH of the reaction solution to be 4.7-4.8 by using 1M HCl aqueous solution during the reaction; adjusting the pH of the reaction solution to 7.0 by using 1M NaOH aqueous solution to terminate the coupling reaction; transferring the reaction solution into a dialysis bag (Shibipure, 8kDa molecular weight cut-off) for dialysis for 3 days, and freeze-drying the purified solution obtained after dialysis to obtain propylene glycol modified sodium hyaluronate, wherein the water content is below 10 percent, and the hyaluronic acid is stored in a nitrogen-filled sealed and light-proof manner.
(2) And (3) oxidation reaction: adding 10.0g of propylene glycol modified sodium hyaluronate obtained in the step (1) into 1000mL of ultrapure water, and magnetically stirring to dissolve the sodium hyaluronate to obtain 1% (w/v) clear transparent solution; adding a solution containing 20.0g NaIO 4 200mL of 1% sodium silicate (Na) 2 SiO 3 ) Reacting the aqueous solution for 24 hours under the condition of keeping out of the sun and magnetic stirring at room temperature, adding a proper amount of glycol and magnetically stirring for 1 hour to terminate the reaction; transferring the reaction solution into a dialysis bag (Shibipure, 8kDa molecular weight cut-off) for dialysis for 3 days, and freeze-drying the purified solution obtained after dialysis to obtain the aldehyde modified sodium hyaluronate, wherein the water content is below 10%, the aldehyde modification rate is 63% by measurement, and the hyaluronic acid is stored in a nitrogen-filled sealed and dark place. The yield was calculated to be 87.1%.
Example 2 preparation of hydrogel from sodium hyaluronate modified by hydroformylation with Small molecule Cross-linker
Dissolving 1.0g of the aldehyde modified sodium hyaluronate prepared in example 1 in 20mL of distilled water to obtain a solution with the concentration of 5.0%; dissolving 120mg of small molecular cross-linking agent cystine hydrochloride in 2mL of distilled water to obtain a cross-linking agent solution; dropwise adding a cross-linking agent solution into the sodium hyaluronate solution subjected to hydroformylation modification, and magnetically stirring at room temperature to react for 2 hours to obtain a gel substance; soaking in distilled water, and shaking for 24 hr to obtain hydrogel.
Example 3 preparation of hydrogel by self-crosslinking of sodium hyaluronate modified by hydroformylation
Dissolving 1.0g of hydrazide modified sodium hyaluronate prepared in preparation example 1 in 50mL of purified water to obtain a solution with the concentration of 2.0%; in addition, 1.0g of the sodium hyaluronate modified by hydroformylation prepared in example 1 was dissolved in 50mL of purified water to obtain a solution with a concentration of 2.0%. Mixing the two solutions, stirring and reacting at room temperature for 1 minute to form a viscous colloidal substance, wherein the viscosity of the viscous colloidal substance continuously increases with the continuous crosslinking reaction, and does not increase after about 20 minutes to obtain hydrogel, wherein the viscosity of the hydrogel is measured to be 2.152 +/-0.007.
Example 4 physicochemical Properties of aldehyde-modified sodium hyaluronate
1.0g of the aldehyde-modified sodium hyaluronate obtained in example 1 and 1.0g of commercially available sodium hyaluronate having a molecular weight equivalent thereto were dissolved in 100mL of ultrapure water to obtain solutions having a concentration of 1% (w/v), and the kinetic viscosities of the obtained solutions were measured at 25 ℃ with a rotational viscometer, and the results are shown in Table 1.
Meanwhile, 1.0g of the aldehyde-modified sodium hyaluronate prepared in example 1 and 1.0g of commercially available sodium hyaluronate having a molecular weight equivalent thereto were taken, and the stability including appearance, water content, viscosity and the like was examined at room temperature and under normal pressure, and the results are shown in table 1.
Table 1 viscosity and stability observations of sodium hyaluronate modified by hydroformylation
Figure BDA0002582369280000121
Example 5 physicochemical Properties of hydrogel made of sodium hyaluronate modified by aldehyde group
20mL of the aldehyde-modified sodium hyaluronate hydrogel obtained in examples 2 and 3 and a commercially available sodium hyaluronate hydrogel were prepared into a solution having a concentration of 0.5% w/v, respectively, and the kinetic viscosity thereof was measured at 25 ℃ by a rotational viscometer, and the results are shown in Table 2. Meanwhile, the stability including appearance, water content, viscosity, etc. was examined at room temperature and under normal pressure, and the results are shown in table 2.
Table 2 viscosity and stability investigation results of aldehyde modified sodium hyaluronate hydrogel
Figure BDA0002582369280000131
Example 6 MTT assay of aldehyde-modified sodium hyaluronate and hydrogel thereof
The aldehyde-modified sodium hyaluronate prepared in example 1 and the hydrogel prepared in examples 2 and 3 were prepared into a series of samples (0, 500, 1000, 2500 mg/mL) with DMEM cell preparation solutions, respectively, and filtered to sterilize. The sample cytotoxicity test was performed on L929 fibroblasts using the MTT method. One group at a concentration of 0 was a blank control group, and the others were experimental groups. L929 cells grown in log phase were seeded into 96-well cell culture plates (cell concentration 8.0X 10) 3 One/well), after the cells adhere to the wall, adding the blank control group solution and the experimental group solution respectively, wherein each well is 100 mu L, and 3 multiple wells are arranged. After 24h of culture, the absorbance at 570nm was measured by the MTT method and the relative survival rate of the cells was calculated.
The calculation formula is as follows: cell survival (%) = (experimental OD average/blank OD average) × 100%
The results show that the cell survival rate of each experimental group is over 95%. The results show that neither the aldehydized modified sodium hyaluronate prepared in example 1 nor the hydrogels prepared in examples 2 and 3 have significant toxicity.
In this patent, the concentration 1M of the aqueous HCl or NaOH solution is 1mol/L, as it is.

Claims (49)

1. An aldehyde modified hyaluronic acid or sodium hyaluronate having the structure shown by the following formula:
Figure FDA0003973537840000011
2. the aldehyde modified hyaluronic acid or sodium hyaluronate according to claim 1, wherein the molecular weight is between 90 and 110kDa.
3. The aldehyde modified hyaluronic acid or sodium hyaluronate according to claim 1, wherein the modification rate is 60-70%.
4. The method for preparing the aldehyde modified hyaluronic acid or sodium hyaluronate according to any one of claims 1 to 3, comprising the following steps:
(1) Dissolving hyaluronic acid or sodium hyaluronate in water, adjusting the pH of the obtained solution to 4.5-5.0, adding EDCI to stir for dissolution, then adding a propylene glycol hydrazide derivative 2,3-dihydroxypropyl-3- ((3-hydrazino-3-oxopropyl) thio) -2-methylpropionate, stirring for reaction at 20-50 ℃ for 0.5-12h, adjusting the pH of the reaction solution to 6.0-8.0 to stop the reaction, dialyzing the reaction solution, and freeze-drying to obtain propylene glycol modified hyaluronic acid or sodium hyaluronate;
(2) Dissolving the propylene glycol modified hyaluronic acid or sodium hyaluronate obtained in the step (1) in water, adding sodium silicate aqueous solution containing sodium periodate, stirring and reacting for 2-72 hours at the temperature of 20-50 ℃ in a dark place, adding ethylene glycol, stirring for 0.2-4 hours to stop the reaction, dialyzing the reaction solution, and freeze-drying to obtain aldehyde modified hyaluronic acid or sodium hyaluronate.
5. The method according to claim 4, wherein the reaction in step (1) is carried out with stirring for 3 to 5 hours.
6. The method according to claim 4, wherein in the step (1), the pH of the reaction solution is controlled to 4.5 to 5.0 during the reaction.
7. The method according to claim 4, wherein the molecular weight of hyaluronic acid or sodium hyaluronate in step (1) is 90-110kDa.
8. The method according to claim 4, wherein the mass/volume concentration of the aqueous solution of hyaluronic acid or sodium hyaluronate in step (1) is 0.5-1.5%.
9. The method according to claim 4, wherein in the step (1), the mass ratio of EDCI to hyaluronic acid or sodium hyaluronate is 1.
10. The method according to claim 4, wherein in the step (1), the propylene glycol hydrazide derivative is further added after the EDCI is added and stirred to be dissolved, and the stirring is continued for 2 to 60 minutes.
11. The method of claim 10, wherein in step (1), the propylene glycol hydrazide derivative is added after stirring for 5 to 30 minutes after the EDCI is added and dissolved by stirring.
12. The method according to claim 4, wherein in the step (1), the mass ratio of the propylene glycol hydrazide derivative to the hyaluronic acid or sodium hyaluronate is 1.
13. The method of claim 4, wherein in step (1), the lyophilization is carried out until the water content is less than 10%.
14. The method according to claim 4, wherein the stirring reaction in the step (2) is carried out for 16 to 32 hours.
15. The method according to claim 4, wherein in the step (2), the reaction is terminated by stirring for 0.5 to 2 hours.
16. The method according to claim 4, wherein the mass/volume concentration of the propylene glycol modified hyaluronic acid or sodium hyaluronate aqueous solution in step (2) is 0.5-1.5%.
17. The preparation method according to claim 4, wherein in the step (2), the mass ratio of the sodium periodate to the propylene glycol modified hyaluronic acid or sodium hyaluronate is 1.
18. The method according to claim 4, wherein in the step (2), the sodium periodate is present in the aqueous sodium silicate solution at a concentration of 8 to 12% by mass/volume.
19. The method of claim 4, wherein in the step (2), the concentration of sodium silicate in the aqueous solution of sodium silicate is 0.2% to 5% by mass/volume.
20. The method of claim 19, wherein in the step (2), the sodium silicate aqueous solution has a mass/volume concentration of 0.5 to 1.5% sodium silicate.
21. The method of claim 4, wherein in step (2), the lyophilization is carried out until the water content is less than 10%.
22. A composition comprising the aldehydized and modified hyaluronic acid or sodium hyaluronate according to any of claims 1 to 3 and a polymer containing an amino group or hydrazide group, wherein the amino group-containing polymer is selected from the group consisting of chitosan, collagen and gelatin, and the hydrazide group-containing polymer is selected from the group consisting of hydrazide modified hyaluronic acid or sodium hyaluronate, hydrazide modified chondroitin sulfate, hydrazide modified heparin, hydrazide modified sodium alginate, hydrazide modified cellulose, hydrazide modified gelatin, hydrazide modified polyethylene glycol and hydrazide modified dendrimer polyamide.
23. The composition of claim 22, further comprising water.
24. The composition of claim 22, wherein the composition comprises a small molecule crosslinker.
25. The composition of any one of claims 22-24, wherein the mass/volume concentration of aldehyde modified hyaluronic acid or sodium hyaluronate is 0.5% to 2.5%.
26. The composition of any one of claims 22 to 24, wherein the mass/volume concentration of the amino or hydrazide group-containing polymer is from 0.5% to 2.5%.
27. The composition according to any one of claims 22 to 24, wherein the mass ratio of the aldehyde modified hyaluronic acid or sodium hyaluronate to the amino or hydrazide group-containing polymer is 1.
28. The composition of any one of claims 22-24, wherein the amino-containing polymer has a molecular weight of 90-110kDa.
29. The composition of any one of claims 22-24, wherein the hydrazide group-containing polymer has a molecular weight of 90-110kDa.
30. A kit comprising the aldehydized modified hyaluronic acid or sodium hyaluronate according to any of claims 1 to 3 and an amino group-or hydrazide group-containing polymer independently selected from the group consisting of chitosan, collagen and gelatin, wherein the hydrazide group-containing polymer is selected from the group consisting of hydrazide modified hyaluronic acid or sodium hyaluronate, hydrazide modified chondroitin sulfate, hydrazide modified heparin, hydrazide modified sodium alginate, hydrazide modified cellulose, hydrazide modified gelatin, hydrazide modified polyethylene glycol and hydrazide modified dendrimer polyamide.
31. The kit of claim 30, further comprising water, independently present.
32. The kit of claim 30, wherein the kit comprises a small molecule cross-linking agent.
33. The kit according to any one of claims 30 to 32, wherein the mass ratio of the aldehyde modified hyaluronic acid or sodium hyaluronate to the amino or hydrazide group-containing polymer is 1.
34. The kit of any one of claims 30 to 32, wherein the amino-containing polymer has a molecular weight of 90 to 110kDa.
35. The kit of any one of claims 30-32, wherein the hydrazide group-containing polymer has a molecular weight of 90-110kDa.
36. A hydrogel obtained by crosslinking the aldehyde-modified hyaluronic acid or sodium hyaluronate according to any one of claims 1 to 3 with an amino or hydrazide group-containing polymer, wherein the amino group-containing polymer is selected from chitosan, collagen and gelatin, and the hydrazide group-containing polymer is selected from hydrazide-modified hyaluronic acid or sodium hyaluronate, hydrazide-modified chondroitin sulfate, hydrazide-modified heparin, hydrazide-modified sodium alginate, hydrazide-modified cellulose, hydrazide-modified gelatin, hydrazide-modified polyethylene glycol and hydrazide-modified dendrimer polyamide.
37. The hydrogel according to claim 36, wherein the mass ratio of the aldehyde modified hyaluronic acid or sodium hyaluronate to the amino or hydrazide group-containing polymer is 1.
38. The hydrogel of claim 36, wherein the amino-containing polymer has a molecular weight of 90-110kDa.
39. The hydrogel of claim 36, wherein the hydrazide group-containing polymer has a molecular weight of 90 to 110kDa.
40. A method of preparing a hydrogel according to any of claims 36 to 39 comprising the steps of: preparing an aqueous solution containing the aldehydized modified hyaluronic acid or sodium hyaluronate according to any one of claims 1 to 3 and the amino or hydrazide group-containing polymer, and reacting at 20 to 50 ℃.
41. The method of claim 40, wherein the mass/volume concentration of the aldehyde modified hyaluronic acid or sodium hyaluronate in the solution is 0.5% to 2.5%.
42. The method of claim 40, wherein the mass/volume concentration of the amino-or hydrazide-group-containing polymer is from 0.5% to 2.5%.
43. A hydrogel obtained by crosslinking the aldehydic modified hyaluronic acid or sodium hyaluronate according to any one of claims 1 to 3 with a small molecule crosslinking agent containing an amino group or a hydrazide group.
44. The hydrogel of claim 43, wherein the amino group-containing small molecule cross-linking agent is selected from the group consisting of small molecule compounds containing two or more primary amines and physiologically acceptable salts thereof.
45. The hydrogel of claim 44, wherein the small molecule compound containing two or more primary amines is selected from the group consisting of cystine, lysine, ethylenediamine, propylenediamine, butylenediamine, pentylenediamine, hexylenediamine, m-phenylenediamine, p-phenylenediamine, and biphenyldiamine.
46. The hydrogel of claim 44, wherein the salt is selected from the group consisting of hydrochloride, sulfate, mesylate, and citrate.
47. The hydrogel of claim 43, wherein the hydrazide group-containing small molecule cross-linking agent is selected from the group consisting of small molecule compounds containing two or more hydrazides.
48. The hydrogel of claim 47, wherein the small molecule compound comprising two or more hydrazides is selected from the group consisting of oxalic dihydrazide, malonic dihydrazide, succinic dihydrazide, glutaric dihydrazide, adipic dihydrazide, phthalic dihydrazide, isophthalic dihydrazide, and terephthalic dihydrazide.
49. A method of preparing a hydrogel according to any of claims 43 to 48 comprising the steps of: preparing an aqueous solution containing the aldehydized modified hyaluronic acid or sodium hyaluronate according to any one of claims 1 to 3 and an aqueous solution containing the small molecule cross-linking agent containing an amino group or a hydrazide group, dropwise adding the aqueous solution containing the small molecule cross-linking agent containing an amino group or a hydrazide group to the aqueous solution containing the aldehydized modified hyaluronic acid or sodium hyaluronate, and stirring at 20 to 50 ℃ until the cross-linking reaction is completed.
CN202010671220.2A 2020-07-13 2020-07-13 Aldehyde modified hyaluronic acid (sodium) and synthesis method and application thereof Active CN113929792B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202010671220.2A CN113929792B (en) 2020-07-13 2020-07-13 Aldehyde modified hyaluronic acid (sodium) and synthesis method and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202010671220.2A CN113929792B (en) 2020-07-13 2020-07-13 Aldehyde modified hyaluronic acid (sodium) and synthesis method and application thereof

Publications (2)

Publication Number Publication Date
CN113929792A CN113929792A (en) 2022-01-14
CN113929792B true CN113929792B (en) 2023-03-10

Family

ID=79273551

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202010671220.2A Active CN113929792B (en) 2020-07-13 2020-07-13 Aldehyde modified hyaluronic acid (sodium) and synthesis method and application thereof

Country Status (1)

Country Link
CN (1) CN113929792B (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114805631A (en) * 2022-03-01 2022-07-29 吉林省奇健生物技术有限公司 High-activity hyaluronic acid-polypeptide conjugate as well as preparation method and application thereof
CN116375905A (en) * 2022-08-18 2023-07-04 杭州协合医疗用品有限公司 Preparation method of aldehyde chitin and derivative thereof

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6630457B1 (en) * 1998-09-18 2003-10-07 Orthogene Llc Functionalized derivatives of hyaluronic acid, formation of hydrogels in situ using same, and methods for making and using same
WO2007001926A2 (en) * 2005-06-24 2007-01-04 Hyperbranch Medical Technology, Inc. Low-swelling hydrogel sealants for wound repair
WO2007102149A2 (en) * 2006-03-07 2007-09-13 Prochon Biotech Ltd. Hydrazido derivatives of hyaluronic acid
EP3156044A1 (en) * 2015-10-16 2017-04-19 Merz Pharma GmbH & Co. KGaA In situ cross-linkable polysaccharide compositions and uses thereof
CN110128682A (en) * 2018-02-02 2019-08-16 华东理工大学 Sulfydryl-cross-link hydrogel material and the preparation method and application thereof
CN110498936A (en) * 2019-07-15 2019-11-26 北京化工大学 A kind of preparation method of Sodium Hyaluronate/sodium alginate injection-type composite hydrogel
CN110522948A (en) * 2019-08-29 2019-12-03 南京工业大学 Injection aquagel and its preparation method and application

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6630457B1 (en) * 1998-09-18 2003-10-07 Orthogene Llc Functionalized derivatives of hyaluronic acid, formation of hydrogels in situ using same, and methods for making and using same
WO2007001926A2 (en) * 2005-06-24 2007-01-04 Hyperbranch Medical Technology, Inc. Low-swelling hydrogel sealants for wound repair
WO2007102149A2 (en) * 2006-03-07 2007-09-13 Prochon Biotech Ltd. Hydrazido derivatives of hyaluronic acid
EP3156044A1 (en) * 2015-10-16 2017-04-19 Merz Pharma GmbH & Co. KGaA In situ cross-linkable polysaccharide compositions and uses thereof
CN110128682A (en) * 2018-02-02 2019-08-16 华东理工大学 Sulfydryl-cross-link hydrogel material and the preparation method and application thereof
CN110498936A (en) * 2019-07-15 2019-11-26 北京化工大学 A kind of preparation method of Sodium Hyaluronate/sodium alginate injection-type composite hydrogel
CN110522948A (en) * 2019-08-29 2019-12-03 南京工业大学 Injection aquagel and its preparation method and application

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Green Synthetic Approach for Photo-Cross-Linkable Methacryloyl Hyaluronic Acid with a Tailored Substitution Degree;Sigen A等;《Biomacromolecules》;20200409;全文 *
微生物多糖透明质酸生物医用水凝胶研究;王笑 等;《卫生研究》;20190731;全文 *
透明质酸及其衍生物药物载体;张伟等;《化学进展》;20061224(第12期);全文 *

Also Published As

Publication number Publication date
CN113929792A (en) 2022-01-14

Similar Documents

Publication Publication Date Title
US11065365B2 (en) Preparation and/or formulation of proteins cross-linked with polysaccharides
JP5657545B2 (en) Method for preparing an injectable hydrogel crosslinked in an injectable container
AU2007296939B9 (en) Hyaluronic acid derivatives obtained via "click chemistry" crosslinking
JPH03502704A (en) Method for producing water-insoluble biocompatible gel
CN113929792B (en) Aldehyde modified hyaluronic acid (sodium) and synthesis method and application thereof
EP1592715A1 (en) Water soluble and biocompatible gels of hyaluronic acid cross-linked with bi-functional l-aminoacids or l-aminoesters
CN112812329B (en) Hydrogel of sulfhydryl modified high molecular compound, preparation method and application thereof
CN115429935B (en) Injectable cross-linked chondroitin sulfate hydrogel and preparation method thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant