CN114716581A - Multiple modified hyaluronic acid derivative and application thereof - Google Patents
Multiple modified hyaluronic acid derivative and application thereof Download PDFInfo
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Abstract
The multiple modified hyaluronic acid derivative can be flexibly and efficiently used for preparing a hyaluronic acid-drug conjugate due to the fact that the multiple modified hyaluronic acid derivative simultaneously has amino and sulfydryl, can realize the coupling of various drugs and hyaluronic acid through sulfydryl and amino, and can also be used for preparing (double) crosslinked hydrogel with a crosslinking agent with sulfydryl reaction activity and/or a crosslinking agent with amino reaction activity. The multiple modified hyaluronic acid derivative has positive prospect in medical application such as targeted antitumor treatment.
Description
Technical Field
The invention relates to the field of medicines, and particularly relates to a multiple modified hyaluronic acid derivative and application thereof.
Background
Hyaluronic Acid (HA), also known as "Hyaluronic Acid", is a linear anionic non-sulfonated mucopolysaccharide formed by alternating beta-1, 4 and beta-1, 3 glycosidic linkages with glucuronic Acid (GlcA) and N-acetylglucosamine (GlcNAc) as disaccharide units, widely distributed in animal and human extracellular matrices, having high content in skin, lung and intestine, and present in synovial fluid, umbilical cord and blood, and is a main component constituting extracellular matrix and mesenchyme, playing important roles in maintaining extracellular matrix structure and regulating intracellular activities. Hyaluronic acid is widely used in the fields of clinical medicine, advanced cosmetics, beauty and plastic, health food and the like due to its unique physicochemical properties and biological functions.
Hyaluronic acid not only has good physicochemical and biological properties such as biocompatibility, degradability, high viscoelasticity, nonimmunity and the like, but also can be combined with receptors (CD44, RHAMM and the like) over-expressed on the surface of tumor cells, so that the capacity of combining with the tumor cells and internalizing the hyaluronic acid is enhanced, and the hyaluronic acid has important regulation effects on the generation of tumor vessels, the tumor metastasis, the tumor invasiveness and the like (Misra and the like, FEBS J, 2011, 278: 1429-1443). Therefore, the hyaluronic acid is used as an anticancer drug delivery carrier in the field of tumor treatment and is continuously one of the hot spots of tumor targeted drug delivery system research (Oh et al, Qiu et al, J Control Release, 2010, 141(1): 2-12; pharmaceutical science report 2013, 48 (9): 1376-.
The hyaluronic acid-drug conjugate is a prodrug prepared by combining a small-molecule antitumor drug with hyaluronic acid through a covalent bond. These covalent bonds are not readily cleavable in blood, but are cleaved by hydrolysis or enzymatic action upon reaching the target site, releasing the drug. The hyaluronic acid-drug conjugate can improve the solubility of the drug, change the distribution and half-life period of the drug in vivo, increase the accumulation in tumor tissues by enhancing the effect of penetration and retention, and better exert the drug effect.
The side chain of the hyaluronic acid has carboxyl and hydroxyl functional groups for coupling reaction, but the hydroxyl group usually needs to have certain nucleophilic reaction activity in a strong alkaline environment, so that not only are the reaction conditions harsh, but also the strong alkaline can cause the hyaluronic acid to undergo significant main chain hydrolysis and fracture; for carboxyl, the coupling reaction with a specific group (such as an amino group) is usually carried out after the carbodiimide is activated, and the reaction is greatly limited; in summary, the research and development of the hyaluronic acid-drug conjugate are restricted by the two functional groups of carboxyl and hydroxyl due to low reaction efficiency, harsh reaction conditions and the like.
Therefore, it is a problem to be solved to improve the coupling reactivity of hyaluronic acid with drugs. The sodium hyaluronate is modified, and a functional group with higher reactivity is introduced, so that the coupling reaction efficiency of hyaluronic acid and a medicament can be effectively improved. The multiple modified hyaluronic acid derivative with various functional groups with high reactivity can perform specific coupling reaction with various medicines, and particularly has advantages. In addition, the multiple modified hyaluronic acid derivatives having various high reactive functional groups are also advantageous in the preparation of hyaluronic acid crosslinked hydrogels. However, at present, no multiple modified hyaluronic acid derivative having a plurality of highly reactive functional groups such as amino groups and thiol groups has been reported.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a multiple modified hyaluronic acid derivative, which simultaneously has side chains containing amino and sulfydryl, has high reactivity, can be simultaneously coupled with various small-molecule antitumor drugs, can be coupled with specific small-molecule antitumor drugs according to requirements, and can be used as a carrier of various antitumor drugs; meanwhile, the derivative can be better applied to preparing hyaluronic acid cross-linked hydrogel.
In order to solve the technical problems, the multiple modified hyaluronic acid derivative provided by the invention simultaneously comprises a sulfydryl-containing side chain and an amino-containing side chain. Compared with the side chain carboxyl and hydroxyl of hyaluronic acid, the amino and the sulfhydryl in the side chain of the hyaluronic acid derivative have better nucleophilic reaction activity, can be directly coupled with various groups, have high reaction efficiency, and can obviously reduce the dosage of expensive micromolecular antitumor drugs; in addition, the reaction of the two functional groups requires mild conditions, and can be carried out in a neutral or weakly alkaline aqueous environment. The side chain containing amino group of the derivative can be generally coupled with N-hydroxysuccinimide ester (NHS ester), aldehyde group, imido ester, pentafluorophenyl ester, hydroxymethyl phosphine and other groups, when the side chain is reacted with NHS ester, a coupling structure of amido bond can be generated, and when the side chain is reacted with imido ester, a coupling structure of amidine bond can be generated. The side chain containing sulfydryl of the derivative can be generally coupled with groups such as maleimide, halogenated acetyl (bromine-, chlorine-or iodine-), pyridine disulfide, thiosulfonate, vinyl sulfone and the like, when the side chain containing sulfydryl is reacted with maleimide and halogenated acetyl to generate a coupling structure of thioether bond, and is reacted with pyridine disulfide to generate a coupling structure of exchange disulfide bond; therefore, the derivative provided by the invention can realize simultaneous coupling of drug molecules with reactivity with amino and drug molecules with reactivity with sulfydryl, and realize combined targeted therapy of various antitumor drugs. Understandably, the antitumor drug can be independently coupled with amino or sulfhydryl according to the requirement to realize the preparation of the specific antitumor drug.
In addition, the derivative has both amino-containing side chains and mercapto-containing side chains, so that the derivative can be simultaneously crosslinked with a crosslinking agent with mercapto reactivity and a crosslinking agent with amino reactivity to prepare a double-crosslinked hydrogel which is more stable and has a flexibly adjustable crosslinking structure; understandably, the crosslinking hydrogel can be prepared by selectively crosslinking by selecting a crosslinking agent reactive with a mercapto group or a crosslinking agent reactive with an amino group according to requirements.
It should be noted that the multiple modified hyaluronic acid derivatives provided by the present application are prepared by chemically modifying the side chain of hyaluronic acid, and the molecular weight of the adopted hyaluronic acid raw material is 1 ten thousand to 1000 ten thousand daltons; preferably, the molecular weight of the hyaluronic acid raw material used is 10-300 ten thousand daltons; more preferably, the molecular weight of the adopted hyaluronic acid raw material is 20-100 ten thousand daltons, and the molecules of the hyaluronic acid raw material have enough disaccharide units (500-2500 units) for chemical modification and cannot cause reduction of chemical modification efficiency due to overhigh molecular weight; and the multiple modified hyaluronic acid derivative provided by the application has the same linear structure with hyaluronic acid.
The side chain hydroxyl of hyaluronic acid generally needs to have certain nucleophilic reactivity in a strong alkaline environment, so that not only are the reaction conditions harsh, but also the strong alkaline can cause the hyaluronic acid to undergo significant backbone hydrolysis and fracture; the chemical modification of the side chain carboxyl groups of hyaluronic acid can be performed under relatively mild conditions. Therefore, preferably, the side chain containing sulfhydryl group of the derivative of the invention is connected with sulfhydryl group through chemical connecting structure via carboxyl of the side chain; the side chain containing amino of the derivative is connected with amino through a chemical connecting structure through carboxyl of the side chain. Understandably, the side chain hydroxyl and carboxyl of the hyaluronic acid can be subjected to various chemical modifications under specific conditions, and the side chain carboxyl is preferably modified by the invention, and various functional groups such as sulfydryl or amino are introduced; and the form of the chemical linking structure is not limited.
Preferably, the thiol-containing side chain of the derivative is of formula I and/or formula II:
wherein HA is a hyaluronic acid residue; r1And R2Each independently selected from one of substituted or unsubstituted alkylene, aromatic group and polyether group; when present, the substituent is selected from an alkyl group, an amide group, an ester group, or an ether group. More preferably, said R1And R2Each independently selected from-CH2CH2-or-CH2CH2CH2-. In the preferred embodiment, the chemical linking structure is preferably an amide bond, such as formula I and formula II, wherein formula II is an amide bond linking structure including a hydrazide structure, it is understood that the amide bond structure is generally stable, and the amide bond has an electron-withdrawing effect, and can enhance the nucleophilic reaction activity of the thiol group to a certain extent, when the amide bond includes the hydrazide structure, the amino group of the hydrazide structure has a better affinity reaction activity, and in addition, the preparation efficiency of the hydrazide linking structure is higher than that of the amide bondThe connection structure is higher; for R1And R2When different structures are chosen, the reactivity of the nucleophilic reaction differs, for example for formula II, when R2＝CH2CH2When the pKa of the mercapto group is 8.87, when R is2＝CH2CH2CH2The pKa of the mercapto group is 9.01. Understandably, the derivative can simultaneously have a plurality of sulfydryl-containing side chains with different chemical structure connection forms, and the nucleophilic reactivity of each side chain is different, so that the derivative has different potential applications.
Preferably, the side chain containing amino group of the derivative is shown as formula III and/or formula IV:
wherein HA is a hyaluronic acid residue; r3And R4Each independently selected from one of substituted or unsubstituted alkylene, aromatic group and polyether group; when present, the substituent is selected from an alkyl group, an amide group, an ester group, or an ether group. More preferably, said R3And R4Each independently selected from-CH2CH2-or-CH2CH2CH2-. In the preferred scheme, the chemical linking structure is preferably an amide bond, such as formula III and formula IV, wherein the formula IV is an amide bond linking structure containing a hydrazide structure; for R3And R4The nucleophilic activity of the compound(s) is different when different structures are selected, for example, the electron donating effect of the nitrogen adjacent to the amide bond results in an amino group (-C (O) NHNH) in the hydrazide functional group2) (pKa usually 3-4) is higher than that of a conventional amino group (-NH)2) (pKa is usually>8) Has higher nucleophilic reaction activity. Understandably, the derivative can simultaneously have a plurality of amino-containing side chains in different chemical structure connection forms, and the nucleophilic reactivity of each side chain is different, so that the derivative has different potential applications.
The multiple modified hyaluronic acid derivative provided by the invention has both amino and sulfydryl, can be flexibly and efficiently used for preparing a hyaluronic acid-drug conjugate, can realize the coupling of various drugs and hyaluronic acid through sulfydryl and amino, and can also prepare (dual) crosslinked hydrogel with a crosslinking agent with reactivity of sulfydryl and/or a crosslinking agent with reactivity of amino. The multiple modified hyaluronic acid derivative has positive prospect in medical application such as targeted antitumor treatment.
Detailed Description
The technical solutions in the present invention will be described clearly and completely below, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention.
The first embodiment is as follows: preparation of multiple modified hyaluronic acid derivative (thiol having amide bond linkage structure and amino having amide bond linkage structure)
Sodium hyaluronate (molecular weight 100 kilodalton) 1g (2.5mmol) is dissolved in 250ml of distilled water at room temperature, 1-hydroxybenzotriazole 0.667g (5.0mmol) is added, 3-Nitro-2-pyridinedithioethylamine (3-Nitro-2-pyridinesulfenylethyl amine)1g (3.74mmol) is added, and the mixture is stirred and dissolved. Then, the pH of the solution was adjusted to 4.75 with 0.5mol/L hydrochloric acid, 1.43g (7.48mmol) of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride was added, and the pH of the solution was maintained at 4.75 by adding 0.5mol/L hydrochloric acid. After reacting at room temperature for 2 hours, the above reaction solution was put into a dialysis tube (molecular weight cut off 3500), dialyzed with a large amount of 0.2 mol/l sodium chloride solution for 2 days, then dialyzed with a large amount of distilled water for 1 day, and finally the solution in the dialysis tube was collected and freeze-dried to obtain a flocculent solid.
The yellow flocculent solid obtained by the freeze-drying was dissolved in 250ml of distilled water at room temperature, 0.667g (5.0mmol) of 1-hydroxybenzotriazole was added, 1.2g (20mmol) of ethylenediamine was added, and the mixture was stirred and dissolved. Then, the pH of the solution was adjusted to 4.75 with 0.5mol/L hydrochloric acid, and 0.715g (3.74mmol) of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride was added to maintain the pH of the solution at 4.75 with 0.5mol/L hydrochloric acid. After 2 hours of reaction at room temperature, the above reaction solution was put into a dialysis tube (molecular weight cut off 3500), dialyzed with a large amount of 0.2 mol/l sodium chloride solution for 2 days, then dialyzed with a large amount of distilled water for 1 day, and finally the solution in the dialysis tube was collected.
To the dialyzed solution, 3g (10.5mmol) of tris (2-carboxyethyl) phosphine hydrochloride was added, and the mixture was stirred and reacted for 4 hours. The above solution was put into a dialysis tube (molecular weight cut off 3500), dialyzed with a large amount of 0.001 mol/l hydrochloric acid and 0.2 mol/l sodium chloride solution for 2 days, and then dialyzed with a large amount of 0.001 mol/l hydrochloric acid solution for 1 day. And finally, collecting the solution in the dialysis tube, and freeze-drying to obtain flocculent solid, namely the multiple modified hyaluronic acid derivative.
The side chain sulfhydryl content of the multiple modified hyaluronic acid derivative of the invention can be detected by an improved Ellman reagent method (Shu et al, Biomacromolecules 2002,3:1304-1311) or hydrogen nuclear magnetic resonance (H-NMR)1H-NMR) (D2O is solvent), and the content is 223 mu mol/g (taking the characteristic methyl absorption peak of acetyl of hyaluronic acid as an internal standard); the content of amino groups in the side chains can be detected by conventional ninhydrin colorimetry (Chen et al, J. Pharma. Anal., 2005, 25: 526-1H-NMR)(D2O as a solvent), and the like (characteristic methyl absorption peak of acetyl group of hyaluronic acid is taken as an internal standard), and the content thereof is 98 μmol/g.
The side chain mercapto group and the side chain amino group of the multiple modified hyaluronic acid derivative of the invention have the following chemical connection structures respectively:
wherein HA is the hyaluronic acid residue in the multiple modified hyaluronic acid derivative.
Example 2: preparation of multiple modified hyaluronic acid derivative (sulfhydryl containing amide bond connecting structure and hydrazide amino containing amide bond connecting structure)
Sodium hyaluronate (molecular weight 50 kilodaltons) 1g (2.5mmol) was dissolved in 250ml of distilled water at room temperature, and 0.667g (5.0mmol) of 1-hydroxybenzotriazole was added, and 3-Nitro-2-pyridinedithioethylamine (3-Nitro-2-pyridinesulfenylethyl amine)1g (3.74mmol) was added, followed by stirring and dissolution. Then, the pH of the solution was adjusted to 4.75 with 0.5mol/L hydrochloric acid, 1.43g (7.48mmol) of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride was added, and the pH of the solution was maintained at 4.75 by adding 0.5mol/L hydrochloric acid. After reacting at room temperature for 2 hours, the above reaction solution was put into a dialysis tube (molecular weight cut off 3500), dialyzed with a large amount of 0.2 mol/l sodium chloride solution for 2 days, then dialyzed with a large amount of distilled water for 1 day, and finally the solution in the dialysis tube was collected and freeze-dried to obtain a flocculent solid.
The yellow flocculent solid obtained by the freeze-drying was dissolved in 250ml of distilled water at room temperature, 0.667g (5.0mmol) of 1-hydroxybenzotriazole was added, 2.92g (20mmol) of succinic dihydrazide was added, and the mixture was stirred and dissolved. Then, the pH of the solution was adjusted to 4.75 with 0.5mol/L hydrochloric acid, 0.024g (0.125mmol) of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride was added, and the pH of the solution was maintained at 4.75 with the addition of 0.5mol/L hydrochloric acid. After 1 hour of reaction at room temperature, the above reaction solution was put into a dialysis tube (molecular weight cut off 3500), dialyzed with a large amount of 0.2 mol/l sodium chloride solution for 2 days, then dialyzed with a large amount of distilled water for 1 day, and finally the solution in the dialysis tube was collected.
To the dialyzed solution was added 3g (10.5mmol) of tris (2-carboxyethyl) phosphine hydrochloride, and the mixture was stirred and reacted for 4 hours. The above solution was put into a dialysis tube (molecular weight cut off 3500), dialyzed with a large amount of 0.001 mol/l hydrochloric acid and 0.2 mol/l sodium chloride solution for 2 days, and then dialyzed with a large amount of 0.001 mol/l hydrochloric acid solution for 1 day. And finally, collecting the solution in the dialysis tube, and freeze-drying to obtain flocculent solid, namely the multiple modified hyaluronic acid derivative.
The thiol content of the multiple modified hyaluronic acid derivative of the invention can be detected by an improved Ellman reagent method (Shu et al, Biomacromolecules 2002,3:1304-1311) or hydrogen nuclear magnetic resonance (H-NMR) method reported by Shu et al1H-NMR)(D2O is solvent), and the like (taking the characteristic methyl absorption peak of acetyl of hyaluronic acid as an internal standard), and the content of the hyaluronic acid is 246 mu mol/g; the amino group content can be determined according to the standardRegular ninhydrin colorimetric (Chen et al, J. Pharma analysis 2005, 25: 526-1H-NMR)(D2O is a solvent), and the content is 108. mu. mol/g (taking a characteristic methyl absorption peak of acetyl of hyaluronic acid as an internal standard).
The side chain mercapto group and the side chain amino group of the multiple modified hyaluronic acid derivative of the invention have the following chemical connection structures respectively:
wherein HA is the hyaluronic acid residue in the multiple modified hyaluronic acid derivative.
Example 3: preparation of multiple modified hyaluronic acid derivative (sulfhydryl containing two amide bond connecting structures and hydrazide amino containing one amide bond connecting structure)
1g (2.5mmol) of sodium hyaluronate (molecular weight 20 kilodaltons) was dissolved in 250ml of distilled water at room temperature, 0.667g (5.0mmol) of 1-hydroxybenzotriazole was added, and then 4.74g (20mmol) of dithiodipropylhydrazide, 5.32g (20mmol) of dithiodibutylhydrazide and 2.92g (20mmol) of succinic dihydrazide were added and dissolved by stirring. Then, the pH of the solution was adjusted to 4.75 with 0.5mol/L hydrochloric acid, 0.072g (0.375mmol) of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride was added, and the pH of the solution was maintained at 4.75 by adding 0.5mol/L hydrochloric acid. After 1 hour at room temperature, 3g (10.5mmol) of tris (2-carboxyethyl) phosphine hydrochloride was added and the reaction was stirred for 4 hours. The above solution was put into a dialysis tube (molecular weight cut off 3500), dialyzed with a large amount of 0.001 mol/l hydrochloric acid and 0.2 mol/l sodium chloride solution for 2 days, and then dialyzed with a large amount of 0.001 mol/l hydrochloric acid solution for 1 day. And finally, collecting the solution in the dialysis tube, and freeze-drying to obtain flocculent solid, namely the multiple modified hyaluronic acid derivative.
The thiol content of the multiple modified hyaluronic acid derivative of the invention can be detected by an improved Ellman reagent method (Shu et al, Biomacromolecules 2002,3:1304-1311) or hydrogen nuclear magnetic resonance (H-NMR) method reported by Shu et al1H-NMR)(D2O as a solvent), and the like (characteristic of methyl group of acetyl group of hyaluronic acid)Peak collection as internal standard) with a content of 196 mu mol/g; the amino content can be detected by conventional ninhydrin colorimetry (Chen et al, J. Pharma analysis 2005, 25: 526-1H-NMR)(D2O is a solvent), and the content thereof is 112. mu. mol/g (with the characteristic methyl absorption peak of acetyl group of hyaluronic acid as an internal standard).
The side chain sulfhydryl and the side chain amino of the multiple modified hyaluronic acid derivative respectively have the following chemical connection structures:
wherein HA is the hyaluronic acid residue in the multiple modified hyaluronic acid derivative.
Example 4: coupling reaction of multiple modified hyaluronic acid derivative and thiol-reactive model compound
2mg of the multiple modified hyaluronic acid derivative prepared in example 1 was dissolved in 2ml of a phosphate buffer (pH7.0), and 1mg of Maleimide-PEG2-Biotin (EZ-Link Maleimide-PEG2-Biotin) (ThermoFisher) was added thereto, followed by stirring for reaction overnight. And purifying the solution after the reaction by a desalting column to remove unreacted reagents, thus obtaining the biotin coupling hyaluronic acid derivative.
Example 5: coupling reaction of multiple modified hyaluronic acid derivative and amino reactive model compound
2mg of the multiple modified hyaluronic acid derivative prepared in example 2 was dissolved in 2ml of a phosphate buffer solution (pH7.0), and 1.5mg of succinimide-PEG 4-Biotin (EZ-Link NHS-PEG4-Biotin) (ThermoFisher) was added thereto, followed by stirring and reacting for 30 minutes. And purifying the solution after the reaction by a desalting column to remove unreacted reagents, thus obtaining the biotin coupling hyaluronic acid derivative.
Example 6: preparation of double-crosslinked hyaluronic acid gel
Polyethylene glycol divinyl sulfoxide (MW3400) (avadin) 33.32mg and N-hydroxysuccinimide-polyethylene glycol-N-hydroxysuccinimide (MW3400) (avadin) 20mg were dissolved in 5ml of phosphate buffer (pH7.4) to obtain a crosslinking agent solution. 100mg of the multiple modified hyaluronic acid derivative prepared in example 3 was dissolved in 5ml of phosphate buffer (pH7.4), and 5ml of the above-mentioned crosslinking agent solution was added thereto, followed by uniform mixing, whereby the solution gradually lost fluidity and formed a double-crosslinked gel.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (10)
1. A multiple modified hyaluronic acid derivative, wherein the derivative comprises both a thiol-containing side chain and an amino-containing side chain.
2. The derivative of claim 1, wherein the thiol-containing side chain of the derivative is linked to the thiol group through a chemical linking structure via the carboxyl group of the side chain;
the side chain containing amino of the derivative is connected with amino through a chemical connecting structure through carboxyl of the side chain.
3. The derivative of claim 2, wherein the thiol-containing side chain of the derivative is of formula I and/or formula II:
wherein HA is a hyaluronic acid residue; r1And R2Each independently selected from one of substituted or unsubstituted alkylene, aromatic group and polyether group; when present, the substituent is selected from an alkyl group, an amide group, an ester group, or an ether group.
4. The derivative of claim 3, wherein R is1And R2Each independently selected from-CH2CH2-or-CH2CH2CH2-。
5. The derivative of claim 2, wherein the side chain containing an amino group of the derivative is of formula III and/or formula IV:
wherein HA is a hyaluronic acid residue; r is3And R4Each independently selected from one of substituted or unsubstituted alkylene, aromatic group and polyether group; when present, the substituent is selected from an alkyl group, an amide group, an ester group, or an ether group.
6. The derivative of claim 5, wherein R is3And R4Each independently selected from-CH2CH2-or-CH2CH2CH2-。
7. The derivative of claim 1, wherein the hyaluronic acid starting material used to synthesize the derivative has a molecular weight of 1-1000 kilodaltons.
8. The derivative of claim 7, wherein the hyaluronic acid material has a molecular weight of 10-300 kilodaltons.
9. Use of a derivative according to any one of claims 1 to 8 for the preparation of hyaluronic acid-drug conjugates in the medical field.
10. Use of a derivative according to any one of claims 1 to 8 for the preparation of a cross-linked hyaluronic acid material.
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| CN101721349A (en) * | 2008-10-16 | 2010-06-09 | 常州百瑞吉生物医药有限公司 | Injectable in-situ crosslinking aquogel and preparation method and application thereof |
| CN103613686A (en) * | 2013-11-12 | 2014-03-05 | 广州市一杰医药科技有限公司 | Preparation method and application of sulfhydryl hyaluronic acid |
| CN111440334A (en) * | 2020-05-26 | 2020-07-24 | 中国科学院长春应用化学研究所 | Injectable hyaluronic acid-based hydrogel and preparation method thereof |
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2022
- 2022-03-25 CN CN202210306737.0A patent/CN114716581A/en active Pending
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|---|---|---|---|---|
| US20050250939A1 (en) * | 1999-02-03 | 2005-11-10 | Mentor Biopolymers Limited | Process for the production of multiple cross-linked hyaluronic acid derivatives |
| CN1468262A (en) * | 2000-10-10 | 2004-01-14 | LG��ѧ��ʽ���� | Crosslinked amide derivatives of hyaluronic acid and manufacturing method thereof |
| CN101200504A (en) * | 2006-12-11 | 2008-06-18 | 上海百瑞吉生物医药有限公司 | Macromolecule thiolated modified derivatives and cross-linking material thereof |
| CN101721349A (en) * | 2008-10-16 | 2010-06-09 | 常州百瑞吉生物医药有限公司 | Injectable in-situ crosslinking aquogel and preparation method and application thereof |
| CN103613686A (en) * | 2013-11-12 | 2014-03-05 | 广州市一杰医药科技有限公司 | Preparation method and application of sulfhydryl hyaluronic acid |
| CN111440334A (en) * | 2020-05-26 | 2020-07-24 | 中国科学院长春应用化学研究所 | Injectable hyaluronic acid-based hydrogel and preparation method thereof |
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