CN114716581A - A kind of multiple modified hyaluronic acid derivative and its application - Google Patents

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

A kind of multiple modified hyaluronic acid derivative and its application

technical field

The invention relates to the field of medicine, in particular to a multi-modified hyaluronic acid derivative and application thereof.

Background technique

Hyaluronic acid (HA), also known as "hyaluronic acid", is a kind of disaccharide composed of glucuronic acid (GlcA) and N-acetylglucosamine (GlcNAc) through β-1, 4 and β- A straight-chain linear anionic non-sulfonated mucopolysaccharide formed by alternately connecting 1,3 glycosidic bonds, widely distributed in the extracellular matrix of animals and humans, with high content in the skin, lungs and intestines, as well as In the fluid, umbilical cord and blood, it is the main component of the extracellular matrix and the interstitium, and plays an important role in maintaining the structure of the extracellular matrix and regulating intracellular activities. Because of its unique physicochemical properties and biological functions, hyaluronic acid is widely used in clinical medicine, advanced cosmetics, cosmetic surgery and health food.

Hyaluronic acid not only has good biocompatibility, degradability, high viscoelasticity and non-immunogenicity and other physicochemical and biological properties, it can also interact with overexpressed receptors (CD44, RHAMM, etc.) on the surface of tumor cells. Combined, it enhances the ability to bind to tumor cells and internalize hyaluronic acid, and has an important regulatory effect on tumor angiogenesis, tumor metastasis and invasiveness (Misra et al., FEBS J, 2011, 278: 1429-1443 ). Therefore, with the help of the tumor targeting properties of hyaluronic acid, it can be used as a carrier material to construct a tumor-targeted drug delivery system, and anti-cancer drugs can be targeted and delivered into tumor cells, so as to better kill tumor cells, so transparent As an anti-cancer drug delivery carrier, lactic acid has been used in the field of tumor therapy, and has continued to become one of the hot spots in the study of tumor-targeted drug delivery systems (Oh et al., Qiu et al., J Control Release, 2010, 141(1): 2-12; Journal of Pharmacy 2013, 48(9): 1376-1382).

Hyaluronic acid-drug conjugates are prodrugs prepared by combining small molecule antitumor drugs with hyaluronic acid through covalent bonds. These covalent bonds are not easily cleaved in blood, but after reaching the target, they are broken by hydrolysis or enzymatic hydrolysis to release the drug. Hyaluronic acid-drug conjugates can improve the solubility of drugs, change the drug distribution and half-life in vivo, increase the accumulation in tumor tissue by enhancing the osmotic retention effect, and exert better drug efficacy.

There are two functional groups of carboxyl and hydroxyl on the side chain of hyaluronic acid for coupling reaction, but for hydroxyl, it usually needs a strong alkaline environment to have a certain nucleophilic reactivity, not only the reaction conditions are harsh, but also the strong alkaline It will also cause hyaluronic acid to undergo significant main chain hydrolysis and breakage; for carboxyl groups, it is usually necessary to couple with specific groups (such as amino groups) after carbodiimide activation, and the reaction is limited. , The two functional groups, carboxyl and hydroxyl, restrict the research and development of hyaluronic acid-drug conjugates due to low reaction efficiency and harsh reaction conditions.

Therefore, improving the coupling reactivity of hyaluronic acid and drugs is a problem that needs to be solved. Modification of sodium hyaluronate to introduce functional groups with higher reactivity can effectively improve the coupling reaction efficiency of hyaluronic acid and drugs. The multiple modified hyaluronic acid derivatives with a variety of highly reactive functional groups can carry out specific coupling reactions with a variety of drugs, which is particularly advantageous. In addition, these multiple modified hyaluronic acid derivatives with various highly reactive functional groups also have advantages in the preparation of hyaluronic acid cross-linked hydrogels. However, at present, multiple modified hyaluronic acid derivatives with a variety of highly reactive functional groups such as amino groups and sulfhydryl groups have not been reported yet.

SUMMARY OF THE INVENTION

The technical problem to be solved by the present invention is to provide a multi-modified hyaluronic acid derivative, which has side chains containing amino groups and sulfhydryl groups at the same time, and has high reactivity. It can be coupled with specific small-molecule anti-tumor drugs as required, and can be used as a carrier for various anti-tumor drugs; at the same time, the derivative can also be better used in the preparation of hyaluronic acid cross-linked hydrogels.

In order to solve the above-mentioned technical problems, the multiple modified hyaluronic acid derivatives provided by the present invention include a side chain containing a sulfhydryl group and a side chain containing an amino group at the same time. Compared with the side chain carboxyl group and hydroxyl group of hyaluronic acid, the amino group and sulfhydryl group in the side chain of the hyaluronic acid derivative provided by the present invention have better nucleophilic reactivity, and can be directly coupled with various groups for coupling reaction, and The reaction efficiency is high, which can significantly reduce the amount of expensive small-molecule antitumor drugs; in addition, the conditions required for the reaction of the two functional groups are mild and can be carried out in a neutral or weakly alkaline aqueous environment. The amino-containing side chain of the derivative can usually be coupled with N-hydroxysuccinimide ester (NHS ester), aldehyde group, imidate, pentafluorophenyl ester, hydroxymethyl phosphine and other groups. When it reacts with NHS ester, it can generate the coupling structure of amide bond, and when it reacts with imidate, it can generate the coupling structure of amidine bond. The thiol-containing side chain of the derivative can usually be carried out with groups such as maleimide, haloacetyl (bromo-, chloro- or iodo-), pyridinedithio, thiosulfonate, vinyl sulfone, etc. Coupling reaction, when it reacts with maleimide and haloacetyl group to generate the coupling structure of sulfide bond, and reacts with pyridine disulfide group to generate the coupling structure of exchanged disulfide bond; therefore, for the present invention provides Derivatives of it can realize the simultaneous coupling of drug molecules reactive with amino groups and drug molecules reactive with sulfhydryl groups, and realize the combined targeted therapy of various anti-tumor drugs. Understandably, antitumor drugs can also be individually coupled with amino groups or sulfhydryl groups according to requirements to achieve the preparation of specific antitumor drugs.

In addition, since the derivative has both an amino group-containing side chain and a thiol group-containing side chain, the derivative can achieve cross-linking with a cross-linking agent reactive with thiol groups and a cross-linking agent reactive with amino groups at the same time to prepare more stable A double-crosslinked hydrogel whose crosslinking structure can be flexibly adjusted; understandably, a crosslinking agent reactive with sulfhydryl groups or a crosslinking agent reactive with amino groups can be selected for selective crosslinking preparation according to requirements. Cross-linked hydrogels.

It should be noted that the multi-modified hyaluronic acid derivatives provided in this application are prepared by chemically modifying the side chains of hyaluronic acid, and the molecular weight of the hyaluronic acid raw materials used is 10,000-10,000,000 dal preferably, the molecular weight of the hyaluronic acid raw material used is 100,000-3 million daltons; more preferably, the molecular weight of the hyaluronic acid raw material used is 200,000-1 million daltons, and the transparent The raw material molecule of uronic acid has enough disaccharide units (500-2500) for chemical modification and will not reduce the chemical modification efficiency due to the high molecular weight; and the multiple modified hyaluronic acid provided in this application Derivatives have the same straight-chain linear structure as hyaluronic acid.

The side chain hydroxyl of hyaluronic acid usually needs to have a certain nucleophilic reactivity in a strong alkaline environment. Not only the reaction conditions are harsh, but the strong alkalinity will also cause the hyaluronic acid to undergo significant main chain hydrolysis and break; hyaluronic acid The chemical modification of the side chain carboxyl groups can be carried out in a relatively mild environment. Therefore, preferably, the sulfhydryl-containing side chain of the derivative of the present invention is connected through a chemical linking structure through the carboxyl group of the side chain and the sulfhydryl group; the amino group-containing side chain of the derivative of the present invention is through the carboxyl group of the side chain and Amino groups are linked by chemical linking structures. Understandably, the side chain hydroxyl and carboxyl groups of hyaluronic acid can undergo various chemical modifications under specific conditions, and the present invention preferably modifies the side chain carboxyl groups to introduce various functional groups such as sulfhydryl or amino groups; The form is not limited.

Preferably, the derivative has a thiol-containing side chain such as formula I and/or formula II:

Wherein, HA is a hyaluronic acid residue; R ₁ and R ₂ are each independently selected from a substituted or unsubstituted alkylene group, an aryl group, and a polyether group; when a substituent exists, the substituent is selected from an alkane group group, amide group, ester group or ether group. More preferably, each of said R ₁ and R ₂ is independently selected from -CH ₂ CH ₂ - or -CH ₂ CH ₂ CH ₂ -. In this preferred solution, 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 comprising a hydrazide structure. It is understood that the amide bond structure is usually relatively stable, and the amide bond has The electron-withdrawing effect can enhance the nucleophilic reactivity of the sulfhydryl group to a certain extent. When the amide bond contains a hydrazide structure, the amino group of the hydrazide structure has better affinity reactivity. In addition, the preparation efficiency of the hydrazide-linked structure is higher than that of the amide-linked structure. higher; for the selection of R ₁ and R ₂ , usually when different structures are selected, their nucleophilic activity is different, for example, for formula II, when R ₂ =CH ₂ CH ₂ , the pKa of the thiol group is 8.87, and when R When ₂ = _CH2CH2CH2 , the _pKa of the mercapto group is _9.01 . Understandably, the derivatives can have a variety of sulfhydryl-containing side chains with different chemical structures at the same time, and the nucleophilic reactivity of each side chain is also different, so that they have different potential applications.

Preferably, the derivative has an amino-containing side chain such as formula III and/or formula IV:

Wherein, HA is a hyaluronic acid residue; R ₃ and R ₄ are each independently selected from a substituted or unsubstituted alkylene group, an aryl group, and a polyether group; when a substituent exists, the substituent is selected from alkane group, amide group, ester group or ether group. More preferably, each of said R ₃ and R ₄ is independently selected from -CH ₂ CH ₂ - or -CH ₂ CH ₂ CH ₂ -. In this preferred embodiment, the chemical connection structure is preferably an amide bond, such as formula III and formula IV, wherein, formula IV is an amide bond connection structure comprising a hydrazide structure; for the selection of R ₃ and R ₄ , usually when choosing different structures , the nucleophilic activity is different, for example, the electron donating effect of the nitrogen in the adjacent amide bond causes the amino group (-C(O)NHNH ₂ ) (pKa usually 3-4) in the hydrazide functional group to be stronger than the conventional amino group (-NH ₂ ) (pKa is usually >8) with higher nucleophilic reactivity. It is understandable that the derivatives can have a variety of amino-containing side chains with different chemical structures at the same time, and the nucleophilic reactivity of each side chain is also different, so that they have different potential applications.

Since the multi-modified hyaluronic acid derivative of the present invention has both amino groups and sulfhydryl groups, it can be flexibly and efficiently used for the preparation of hyaluronic acid-drug conjugates, and can realize the coupling of various drugs and hyaluronic acid through sulfhydryl groups and amino groups at the same time. , and at the same time, a (dual) cross-linked hydrogel can be prepared with a thiol-reactive cross-linking agent and/or an amino-reactive cross-linking agent. The multi-modified hyaluronic acid derivatives of the present invention have positive prospects in medical applications such as targeted anti-tumor therapy.

Detailed ways

The technical solutions in the present invention will be described clearly and completely below. Obviously, the described embodiments are a part of the embodiments of the present invention, but not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the protection scope of the present invention.

Example 1: Preparation of multiple modified hyaluronic acid derivatives (containing a sulfhydryl group with an amide bond structure and an amino group with an amide bond structure)

1 g (2.5 mmol) of sodium hyaluronate (molecular weight 1 million Daltons) was dissolved in 250 ml of distilled water at room temperature, 0.667 g (5.0 mmol) of 1-hydroxybenzotriazole was added, and 3-nitro-2-pyridinedithiol was added. 1 g (3.74 mmol) of 3-Nitro-2-pyridinesulfenylethylamine was dissolved by stirring. Then the pH value of the solution was adjusted to 4.75 with 0.5mol/L hydrochloric acid, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride 1.43g (7.48mmol) was added, 0.5mol/L L hydrochloric acid kept the pH of the solution at 4.75. After 2 hours of reaction at room temperature, the above-mentioned reaction solution was loaded into a dialysis tube (molecular weight cut-off 3500), dialyzed with a large amount of 0.2 mol/liter sodium chloride solution for 2 days, then dialyzed with a large amount of distilled water for 1 day, and finally collected in the dialysis tube. The solution was freeze-dried to obtain a flocculent solid.

The yellow flocculent solid obtained by freeze-drying was dissolved in 250 ml of distilled water at room temperature, 0.667 g (5.0 mmol) of 1-hydroxybenzotriazole was added, 1.2 g (20 mmol) of ethylenediamine was added, and the mixture was stirred and dissolved. Then the pH value of the solution was adjusted to 4.75 with 0.5mol/L hydrochloric acid, 0.715g (3.74mmol) of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride was added, and 0.5mol/L L hydrochloric acid kept the pH of the solution at 4.75. After 2 hours of reaction at room temperature, the above-mentioned reaction solution was loaded into a dialysis tube (molecular weight cut-off 3500), dialyzed with a large amount of 0.2 mol/liter sodium chloride solution for 2 days, then dialyzed with a large amount of distilled water for 1 day, and finally collected in the dialysis tube. The solution.

To the solution after dialysis, 3 g (10.5 mmol) of tris(2-carboxyethyl) phosphine hydrochloride was added, and the reaction was stirred for 4 hours. The above solution was loaded 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 sky. Finally, the solution in the dialysis tube is collected, and the flocculent solid obtained by freeze-drying is the multi-modified hyaluronic acid derivative of the present invention.

The content of the side chain sulfhydryl group of the above-mentioned multiple modified hyaluronic acid derivatives ^of the present invention can be detected by the improved Ellman reagent method reported by Shu et al. NMR) (D ₂ O is the solvent) and other methods (using the characteristic methyl absorption peak of the acetyl group of hyaluronic acid as the internal standard), its content is 223 μmol/g; (Chen et al., Journal of Pharmaceutical Analysis, 2005, 25: 526-529) or hydrogen spectroscopic nuclear magnetic resonance detection ( ¹ H-NMR) (D ₂ O as solvent) and other methods (using the characteristic methyl of the acetyl group of hyaluronic acid) The base absorption peak is the internal standard), and its content is 98 μmol/g.

The side chain sulfhydryl group and the side chain amino group of the above-mentioned multiple modified hyaluronic acid derivatives of the present invention respectively have the following chemical connection structures:

Wherein HA is the hyaluronic acid residue in the multiple modified hyaluronic acid derivatives.

Example 2: Preparation of multiple modified hyaluronic acid derivatives (containing a sulfhydryl group with an amide bond structure and a hydrazide amino group with an amide bond structure)

1 g (2.5 mmol) of sodium hyaluronate (molecular weight 500,000 Daltons) was dissolved in 250 ml of distilled water at room temperature, 0.667 g (5.0 mmol) of 1-hydroxybenzotriazole was added, and 3-nitro-2-pyridinedithiol was added. 1 g (3.74 mmol) of 3-Nitro-2-pyridinesulfenylethylamine was dissolved by stirring. Then the pH value of the solution was adjusted to 4.75 with 0.5mol/L hydrochloric acid, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride 1.43g (7.48mmol) was added, 0.5mol/L L hydrochloric acid kept the pH of the solution at 4.75. After 2 hours of reaction at room temperature, the above-mentioned reaction solution was loaded into a dialysis tube (molecular weight cut-off 3500), dialyzed with a large amount of 0.2 mol/liter sodium chloride solution for 2 days, then dialyzed with a large amount of distilled water for 1 day, and finally collected in the dialysis tube. The solution was freeze-dried to obtain a flocculent solid.

The yellow flocculent solid obtained by freeze-drying was dissolved in 250 ml of distilled water at room temperature, 0.667 g (5.0 mmol) of 1-hydroxybenzotriazole was added, 2.92 g (20 mmol) of succinic dihydrazide was added, and the mixture was stirred and dissolved. Then the pH value 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 0.5mol/L L hydrochloric acid kept the pH of the solution at 4.75. After 1 hour of reaction at room temperature, the above reaction solution was loaded into a dialysis tube (molecular weight cut-off 3500), dialyzed with a large amount of 0.2 mol/liter sodium chloride solution for 2 days, and then dialyzed with a large amount of distilled water for 1 day, and finally collected the inner diameter of the dialysis tube. The solution.

To the solution after dialysis, 3 g (10.5 mmol) of tris(2-carboxyethyl) phosphine hydrochloride was added, and the reaction was stirred for 4 hours. The above solution was loaded 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 sky. Finally, the solution in the dialysis tube is collected, and the flocculent solid obtained by freeze-drying is the multi-modified hyaluronic acid derivative of the present invention.

The sulfhydryl content of the above-mentioned multiple modified hyaluronic acid derivatives ^of the present invention can be detected by the improved Ellman reagent method reported by Shu et al. (D ₂ O is the solvent) and other methods (using the characteristic methyl absorption peak of the acetyl group of hyaluronic acid as the internal standard), its content is 246 μmol/g; the amino content can be determined by conventional ninhydrin colorimetry (Chen et al. , Journal of Pharmaceutical Analysis, 2005, 25: 526-529) or hydrogen spectrum nuclear magnetic resonance detection ( ¹ H-NMR) (D ₂ O is the solvent) and other methods (the characteristic methyl absorption peak of the acetyl group of hyaluronic acid is Internal standard), its content was 108 μmol/g.

The side chain sulfhydryl group and the side chain amino group of the above-mentioned multiple modified hyaluronic acid derivatives of the present invention respectively have the following chemical connection structures:

Wherein HA is the hyaluronic acid residue in the multiple modified hyaluronic acid derivatives.

Example 3: Preparation of Multiple Modified Hyaluronic Acid Derivatives (Containing two sulfhydryl groups connected by amide bonds and one hydrazide amino group connected by amide bonds)

1 g (2.5 mmol) of sodium hyaluronate (molecular weight 200,000 Daltons) was dissolved in 250 ml of distilled water at room temperature, 0.667 g (5.0 mmol) of 1-hydroxybenzotriazole was added, and then 4.74 g of dithiodipropionhydrazide ( 20 mmol), 5.32 g (20 mmol) of dithiodibutyric hydrazide, and 2.92 g (20 mmol) of succinic acid dihydrazide, and were dissolved by stirring. Then the pH value 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 0.5mol/L L hydrochloric acid kept the pH of the solution at 4.75. After 1 hour of reaction at room temperature, 3 g (10.5 mmol) of tris(2-carboxyethyl)phosphine hydrochloride was added, and the reaction was stirred for 4 hours. The above solution was loaded 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 sky. Finally, the solution in the dialysis tube is collected, and the flocculent solid obtained by freeze-drying is the multi-modified hyaluronic acid derivative of the present invention.

The sulfhydryl content of the above-mentioned multiple modified hyaluronic acid derivatives ^of the present invention can be detected by the improved Ellman reagent method reported by Shu et al. (D ₂ O is the solvent) and other methods (using the characteristic methyl absorption peak of the acetyl group of hyaluronic acid as the internal standard), its content is 196 μmol/g; the amino content can be determined by conventional ninhydrin colorimetry (Chen et al. , Journal of Pharmaceutical Analysis, 2005, 25: 526-529) or hydrogen spectrum nuclear magnetic resonance detection ( ¹ H-NMR) (D ₂ O is the solvent) and other methods (the characteristic methyl absorption peak of the acetyl group of hyaluronic acid is Internal standard), its content was 112 μmol/g.

The side chain sulfhydryl group and the side chain amino group of the above-mentioned multiple modified hyaluronic acid derivatives of the present invention respectively have the following chemical connection structures:

Wherein HA is the hyaluronic acid residue in the multiple modified hyaluronic acid derivatives.

Example 4: Coupling reaction of multiple modified hyaluronic acid derivatives with thiol-reactive model compounds

Take 2mg of the multi-modified hyaluronic acid derivative prepared in Example 1 and dissolve it in 2ml of phosphate buffer (pH7.0), add 1mg of maleimide-PEG2-biotin (EZ-Link Maleimide-PEG2-Biotin) (ThermoFisher), and the reaction was stirred overnight. After the reaction, the solution is purified by a desalting column to remove unreacted reagents, so as to obtain a biotin-coupled hyaluronic acid derivative.

Example 5: Coupling Reaction of Multiple Modified Hyaluronic Acid Derivatives with Amino-Reactive Model Compounds

Dissolve 2 mg of the multi-modified hyaluronic acid derivative prepared in Example 2 in 2 ml of phosphate buffer (pH 7.0), add 1.5 mg of succinimide-PEG4-biotin (EZ-Link NHS-PEG4-Biotin) (ThermoFisher), and the reaction was stirred for 30 minutes. After the reaction, the solution is purified by a desalting column to remove unreacted reagents, so as to obtain a biotin-coupled hyaluronic acid derivative.

Example 6: Preparation of double cross-linked hyaluronic acid gel

Polyethylene glycol divinyl sulfoxide (MW3400) (Aladdin) 33.32mg and N-hydroxysuccinimide-polyethylene glycol-N-hydroxysuccinimide (MW3400) (Aladdin) 20mg dissolved in 5ml Phosphate buffer (pH 7.4) to obtain a crosslinker solution. Dissolve 100 mg of the multi-modified hyaluronic acid derivative prepared in Example 3 in 5 ml of phosphate buffer (pH 7.4), then add 5 ml of the above cross-linking agent solution, mix evenly, the solution gradually loses fluidity and forms double cross-linking gel.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the present invention. within the scope of protection.

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.