Multifunctional gamma-PGA carrier material as well as preparation method and application thereof

Abstract

The invention discloses a multifunctional gamma-PGA carrier material as well as a preparation method and application thereof, and belongs to the technical field of biological carrier materials. Comprising the following steps: S1, taking a proper amount of beta cyclodextrin and paratoluensulfonyl chloride, dissolving the beta cyclodextrin and the paratoluensulfonyl chloride in a NaOH solution, violently stirring under the reaction condition of 0 DEG C, and performing post-treatment to obtain beta-CD-TS; s2, adding the compound beta-CD-TS obtained in the step S1 into a proper amount of ethidene diamine for reaction, and performing post-treatment to obtain a compound beta-CD-NH2; and S3, activating the beta-CD-NH2 obtained in the step S2, 3-aminophenylboronic acid and 4-dimethylaminopyridine (DMAP) in a reaction bottle by using an N, N-dimethylformamide (DMF) solution, simultaneously taking a proper amount of gamma-PGA and a proper amount of dicyclohexylcarbodiimide (DCC00) in the reaction bottle, activating by using DMF as a solvent, and merging liquids to continue the reaction to obtain a product. The gamma-PGA molecular structure prepared by the invention has high drug loading rate and good physicochemical properties, and can be used in the aspects of drug sustained release, wound tissue repair, biological adhesion, hemostatic, enzyme packaging and the like.

Description

Technical Field

[0001] The invention relates to the technical field of biological carrier materials, and in particular to a multi-functionalized γ-PGA carrier material and a preparation method and application thereof. Background Art

[0002] Polyglutamic acid (γ-PGA), also known as natto gum and polyglutamic acid, is a biopolymer produced by microbial fermentation or extracted from natto. γ-PGA is a special anionic homopolyamide, which is a homopolyamide γ-PGA formed by amide linkage between α-amino and γ-carboxyl. γ-PGA has been a hot topic in drug carrier research due to its biocompatibility and biodegradability, and the carboxyl groups on the side chains can provide polymerization sites for drug molecules. For example, γ-PGA is used as a drug carrier loaded into nanoparticles to achieve antibacterial, immunomodulatory and sustained treatment of oral ulcers (Int J Pharm. 2023, 648: 123607.); γ-PGA itself also has therapeutic effects. γ-PGA helps improve insulin resistance, relieves liver inflammation, and enhances the activity of liver antioxidant enzymes (Int J Biol Macromol. 2024, 261: 129809.).

[0003] In addition, a series of derivatives prepared based on γ-PGA, including various types of polymers such as copolymers, grafted polymers and dendrimers, further enhance the loading capacity of γ-PGA and broaden its application. For example, γ-PGA is subjected to an amide reaction with amino acid ethyl ester hydrochloride (such as L-valine, L-leucine, L-methionine, L-tryptophan or L-phenylalanine, etc.) to obtain amino acid ethyl ester-modified γ-PGA (γ-PGA-graft-AAE), and phenylalanine-modified γ-PGA (γ-PGA-Phe) formed nanoparticles loaded with polypeptide vaccines can produce acquired immune anti-tumor therapeutic effects on mouse liver cancer cells MC38 (Colloid Polym Sci, 2014, 292: 2663-71). However, when the existing γ-PGA is used as a drug carrier material, the drug loading capacity is relatively limited, and the drug loading amount is low. In view of the above problems, the present invention proposes a multi-functionalized γ-PGA carrier material and its preparation method and application. Summary of the invention

[0004] The purpose of the present invention is to provide a multifunctional γ-PGA carrier material and a preparation method and application thereof, so as to solve the problem of low drug loading capacity of the existing γ-PGA mentioned in the above background technology. The prepared γ-PGA molecular structure has a high drug loading rate and good physical and chemical properties, and can be used for drug sustained release, wound tissue repair, bioadhesion, hemostatic agent, enzyme packaging and other aspects.

[0005] To achieve the above object, the present invention provides a multifunctional γ-PGA carrier material, the chemical structure of the multifunctional γ-PGA carrier material is: ; Among them, m is 150-350 and n is 6-8.

[0006] The present invention also provides a method for preparing the above-mentioned multi-functionalized γ-PGA carrier material, which specifically comprises the following steps: S1. Take β-cyclodextrin and p-toluenesulfonyl chloride, dissolve them in NaOH solution, stir to react, and post-treat to obtain compound C 49 H7O 37 S, chemical structure formula: ; S2, compound C obtained in step S1 49 H7O 37 S is added to ethylenediamine for reaction, and then post-processed to obtain compound C 48 H 84 N2O 34 , the chemical structure is: ; S3, compound C obtained in step S2 48 H 84 N2O 34 3-aminophenylboronic acid and 4-dimethylaminopyridine were placed in a reaction bottle and activated with DMF solvent. Meanwhile, γ-PGA and DCC were placed in another reaction bottle and activated with DMF solvent. The liquids in the two reaction bottles were mixed for reaction and post-processed to obtain the target product.

[0007] Preferably, in step S1, the molar mass ratio of β-cyclodextrin, p-toluenesulfonyl chloride and NaOH is one of 1:40:5, 1:45:7 or 1:50:10; the reaction temperature is 0°C, and the stirring reaction time is 60-120 min.

[0008] Preferably, in step S1, the post-treatment method is: filtering the solution after the reaction, taking the filtrate, adjusting the pH to 2-5, standing for 8-24 hours to obtain a precipitate, filtering and drying to obtain the amination product C 49 H7O 37 S.

[0009] Preferably, in step S2, C 49 H7O 37 The molar mass ratio of S to ethylenediamine is one of 1:50, 1:100, 1:180 or 1:200; the reaction temperature is 30-60° C., and the reaction time is 24-72 hours.

[0010] Preferably, in step S2, the post-treatment method is: pouring the reaction solution into a sufficient amount of acetone to obtain a precipitate, and filtering to obtain the product C 48 H 84 N2O 34 .

[0011] Preferably, in step S3, 3-aminophenylboronic acid, C 48 H 84 N2O 34 The molar mass ratio of γ-PGA to DMAP is 6:1:50, 6:1:100 or 6:1:150; the mass ratio of γ-PGA to DCC is 1:1-4; the activation time is 60-120 minutes; the mixing reaction time is 24-72 hours, and the mixing reaction temperature is 40-80°C.

[0012] Preferably, in step S3, the post-treatment method is: drop the reacted solution into ice water, filter to obtain a filtrate, add ethyl acetate to the filtrate to extract until the upper liquid is clarified, take the lower solution, concentrate the lower liquid and drop it into acetone to precipitate, and filter to obtain the target product.

[0013] The present invention also provides application of the multi-functionalized γ-PGA carrier material in carrying drug molecules.

[0014] Therefore, the present invention provides a multi-functionalized γ-PGA carrier material and a preparation method and application thereof, which have the following beneficial effects: (1) The present invention uses γ-PGA as the main chain, and adopts a one-pot method to graft cyclodextrin derivatives and phenylboronic acid derivatives onto γ-PGA to prepare a multifunctional polymer nano drug-carrying material with biodegradable γ-polyglutamic acid as the main chain and cyclodextrin β-CD and 3-aminophenylboronic acid as side chains; (2) The carboxyl group of γ-polyglutamic acid can load ions on the one hand, and on the other hand, it can be used to modify functional groups. The carrier material can be biodegraded into glutamic acid in the body; the β-CD cavity can encapsulate ions, drug molecules, etc.; aminophenylboronic acid is fluorescent, so its fluorescence can be used to characterize compounds. At the same time, it has a boron hydroxyl group that can complex ions; (3) γ-PGA can cross-link itself, cross-link with other compounds, and complex with metal particles in a variety of ways, significantly changing the molecular structure of γ-PGA so that it has a high drug loading rate and good physical and chemical properties. It can be used for drug sustained release, wound tissue repair, bioadhesion, hemostatic agents, enzyme packaging, etc.

[0015] The technical solution of the present invention is further described in detail below through the accompanying drawings and embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The nuclear magnetic resonance hydrogen spectrum (DMSO- d 6, 500M, 1 H); Figure 2 This is a high-load scanning electron microscopy image of the γ-PGA carrier material prepared in Example 5 loaded with the anti-tumor drug doxorubicin; Figure 3 This is a high-load scanning electron microscopy image of the γ-PGA carrier material prepared in Example 8 loaded with the anti-tumor drug doxorubicin; Figure 4 This is a high-load scanning electron microscopy image of the γ-PGA carrier material prepared in Example 11 loaded with the anti-tumor drug doxorubicin; Figure 5 This is a high-load scanning electron microscopy image of the γ-PGA carrier material prepared in Example 13 loaded with the anti-tumor drug doxorubicin; Figure 6 This is a high-load scanning electron micrograph of the γ-PGA carrier material prepared in Example 16 loaded with the anti-tumor drug doxorubicin. DETAILED DESCRIPTION

[0017] The present invention provides a method for preparing a polymer nano drug-carrying material by using γ-PGA as the main chain and grafting β-cyclodextrin and 3-aminophenylboronic acid onto γ-PGA in a one-pot method. The specific steps are as follows: S1: Take appropriate amounts of β-cyclodextrin and p-toluenesulfonyl chloride, dissolve them in NaOH solution, stir vigorously at 0°C, and post-treat to obtain compound C 49 H7O 37 S; S2 Compound C obtained in step S1 49 H7O 37 S is added to an appropriate amount of ethylenediamine for reaction, and then post-processed to obtain compound C 48 H 84 N2O 34 ; S3 Compound C obtained in step S2 48 H 84 N2O34 Activate with 3-aminophenylboronic acid and 4-dimethylaminopyridine (DMAP) in a reaction bottle with N,N-dimethylformamide (DMF) solution. Meanwhile, take an appropriate amount of γ-PGA and an appropriate amount of dicyclohexylcarbodiimide (DCC00) in a reaction bottle with DMF as solvent for activation. Combine the liquids and continue the reaction to obtain the product.

[0018] In the present invention, the selected γ-polyglutamic acid (CAS No. 25513-46-6) has a structural formula as shown in formula (1): ; Its monomer structure is shown in formula (2): ; (2) In the present invention, the selected β-cyclodextrin (β-CD) (CAS No. 7585-39-9) is environmentally friendly and has a non-polar (hydrophobic) cavity and a hydrophilic exterior. They can form inclusion complexes with guest compounds in a reversible manner. The interaction between the host and the guest can effectively capture a variety of bioactive compounds, which is helpful for the delivery and absorption of drugs. Both polar and non-polar organic molecules can be used as guests and can carry the required drugs through the inclusion complex of cyclodextrin, so they can be used as a good drug carrier. Its structural formula is shown in formula (3): ; In the present invention, the selected 3-aminophenylboronic acid (CAS No. 206658-89-1) can be used as a fluorescent probe for active oxygen bond breaking. The fluorescence change effect of the compound produced by the complexation with Alizarin Red can be used to determine the breaking and complexation of chemical bonds. Its structural formula is shown in Formula (4): .

[0019] In the present invention, in step S1, the optimal molar mass ratio of β-cyclodextrin, p-toluenesulfonyl chloride and NaOH is 1:40:5, followed by 1:45:7, and then 1:50:10; the stirring reaction time is 60-120 min.

[0020] The post-treatment method is as follows: filter the solution after the reaction to obtain the filtrate, adjust the pH to 2-5, the best pH is 2, the second best pH is 3, and the third best pH is 4-5; let it stand for 8-24 hours to obtain a precipitate, 12 hours is appropriate, 8 hours is second, and 24 hours is optimal; after filtering and drying, the first step product of amination, β-CD-TS, is obtained.

[0021] In the present invention, in step S2, C 49 H7O 37The molar mass ratio of S to ethylenediamine is 1:50, 1:100, 1:180, and 1:200. The reaction temperature is 30°C-60°C, 30°C is suitable, 60°C is the second, and 40°C is the best. The reaction time is 24-72h. In step S2, the post-treatment method is: take the solution after the reaction, pour it into a sufficient amount of acetone to obtain a precipitate, and filter to obtain the second-step product β-CD-NH2 (the product easily absorbs water, and it is recommended to collect the product by centrifugation).

[0022] In the present invention, in step S3, 3-aminophenylboronic acid, C 48 H 84 N2O 34 The molar mass ratio of γ-PGA to DMAP is 6:1:50, 6:1:100 is the best, and 6:1:150 is the second best. The mass ratio of γ-PGA:DCC is 1:1-4, among which 1:2 is suitable, 1:4 is the best, and 1:3 is the second best. The activation time is 60-120min, among which 120min is the best, and 60min is the second best. The combined liquid reaction time is 24-72h, among which 24h is suitable, 48h is the best, and there is no obvious improvement after 72h. The reaction temperature is 40-80℃, among which 30℃ is suitable, 60℃ is the best, and 80℃ is the second best.

[0023] The post-treatment method is: take the solution after the reaction, drop it into ice water, filter to obtain the filtrate, add a large amount of ethyl acetate (EA) to the filtrate and extract for more than 3 times until the upper liquid is clear, take the lower solution, concentrate the lower liquid and drop it into a large amount of acetone to precipitate to obtain the product.

[0024] The technical solution of the present invention is further described below in conjunction with specific drawings and embodiments. It should be understood that these embodiments are only used to illustrate the present invention and are not used to limit the scope of the present invention. Any other changes, modifications, substitutions, combinations, and simplifications made without violating the spirit and principle of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention. In addition, it should be understood that after reading the content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application, and all belong to the scope of protection of the present invention.

[0025] Reference to "embodiment" herein means that the specific features, structures or characteristics described in conjunction with the embodiment may be included in at least one embodiment of the present application. The term "embodiment" appearing in various places in the specification does not necessarily refer to the same embodiment, nor does it particularly limit its independence or association with other embodiments. In principle, in the present application, as long as there is no technical contradiction or conflict, the various technical features mentioned in the embodiments can be combined in any way to form a corresponding implementable technical solution.

[0026] Unless otherwise defined, the technical terms used in this document have the same meanings as those generally understood by those skilled in the art to which this application belongs; the use of relevant terms in this document is only for describing specific embodiments and is not intended to limit this application.

[0027] Unless otherwise specified in the present invention, the reagents, instruments, equipment, etc. used in the following examples are all reagents, instruments, and equipment commonly used by technicians in this field.

[0028] The final compounds prepared in the following examples have different degrees of substitution, please refer to Figure 1 The molecular weight distribution of γ-PGA itself is between 20,000 and 50,000. m is not a fixed value, but a range, so the structure is not single, but a mixture of different molecular weights. The NMR images provided in this article are only used to illustrate the chemical shift of the functional groups obtained by synthesis, so one spectrum can be used to illustrate it.

[0029] Example 1 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material, which specifically comprises the following steps: S1. Take 4.0 g of β-cyclodextrin and 3.0 g of p-toluenesulfonyl chloride, dissolve them in 50 mL of 0.5 NaOH solution, stir vigorously at 0°C for 60 min, filter the solution after the reaction to obtain the filtrate, adjust the pH to 2, let it stand for 24 h to obtain a precipitate, filter and dry to obtain the first step product of amination, β-CD-TS (compound C 49 H7O 37 S), the chemical structure is: ; S2, 1.5 g of the compound obtained in step S1 58 H 93 O 43 S was added to 40 mL of ethylenediamine and reacted at 40°C for 48 h. The solution after the reaction was taken and poured into a sufficient amount of acetone to obtain a precipitate. The second step product β-CD-NH2 (C 48 H 84 N2O 34), the chemical structure is: ; S3, weigh 2g DMAP, 1g 3-aminophenylboronic acid, 0.2g β-CD-NH2, dissolve in 50ml DMF, 500rt, 40℃ stirring for 1h; weigh 2g γ-PGA, 4.95g DCC, dissolve in 50ml DMF, 500rt, 40℃ stirring for 1h; combine the reaction solutions and continue to react for 24h. After the reaction is completed, drop it into 600ml ice water, precipitate, filter to obtain filtrate, concentrate the filtrate to 40ml, transfer the liquid and add 200ml EA to extract, obtain the lower layer liquid, continue to extract the lower layer liquid twice, drop the extracted lower layer solution into 200ml acetone, and obtain the final product, which is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0030] Example 2 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 1, specifically: Steps S1 and S2 are the same as those in Example 1 and will not be repeated here.

[0031] In step S3, DMAP and 3-aminophenylboronic acid β-CD-NH2 are dissolved in DMF and stirred at 35° C. for 2 h; γ-PGA and DCC are dissolved in DMF and stirred at 40° C. for 2 h.

[0032] The rest is the same as in Example 1, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material. Figure 1 As shown by Figure 1 It can be seen that the hydrogen H9δ of the amide bond formed by the bonding of β-cyclodextrin and γ-polyglutamic acid is 8.0ppm, and the H9δ formed by the amide reaction of 3-aminophenylboronic acid and γ-polyglutamic acid is 8.0ppm. e δ = 9.7ppm, aromatic hydrogen H of 3-aminophenylboronic acid 1,2,3,4 δ = 7-8ppm.

[0033] Example 3 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 1, specifically: Steps S1 and S2 are the same as those in Example 1 and will not be repeated here.

[0034] After the reaction solutions are combined in step S3, the reaction is continued for 48 hours.

[0035] The rest is the same as in Example 1, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0036] Example 4 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 1, specifically: Steps S1 and S2 are the same as those in Example 1 and will not be repeated here.

[0037] In step S3, DMAP and 3-aminophenylboronic acid β-CD-NH2 are dissolved in DMF and stirred at 40° C. for 2 h; γ-PGA and DCC are dissolved in DMF and stirred at 40° C. for 2 h; after the reaction solutions are combined, the reaction is continued for 48 h.

[0038] The rest is the same as in Example 1, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0039] Example 5 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 1, specifically: Steps S1 and S2 are the same as those in Example 1 and will not be repeated here.

[0040] In step S3, DMAP and 3-aminophenylboronic acid β-CD-NH2 are dissolved in DMF and stirred at 80° C. for 1 h; γ-PGA and DCC are dissolved in DMF and stirred at 80° C. for 1 h.

[0041] The rest is the same as in Example 1, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0042] Example 6 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 1, specifically: Steps S1 and S2 are the same as those in Example 1 and will not be repeated here.

[0043] In step S3, DMAP and 3-aminophenylboronic acid β-CD-NH2 are dissolved in DMF and stirred at 80° C. for 2 h; γ-PGA and DCC are dissolved in DMF and stirred at 80° C. for 2 h.

[0044] The rest is the same as in Example 1, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0045] Example 7 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 1, specifically: Steps S1 and S2 are the same as those in Example 1 and will not be repeated here.

[0046] In step S3, DMAP and 3-aminophenylboronic acid β-CD-NH2 are dissolved in DMF and stirred at 80° C. for 1 h; γ-PGA and DCC are dissolved in DMF and stirred at 80° C. for 1 h; after the reaction solutions are combined, the reaction is continued for 48 h.

[0047] The rest is the same as in Example 1, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0048] Example 8 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 1, specifically: Steps S1 and S2 are the same as those in Example 1 and will not be repeated here.

[0049] In step S3, DMAP and 3-aminophenylboronic acid β-CD-NH2 are dissolved in DMF and stirred at 40° C. for 2 h; γ-PGA and DCC are dissolved in DMF and stirred at 40° C. for 2 h; after the reaction solutions are combined, the reaction is continued for 48 h.

[0050] The rest is the same as in Example 1, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0051] Example 9 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 1, specifically: Steps S1 and S2 are the same as those in Example 1 and will not be repeated here.

[0052] In step S3, DMAP and 3-aminophenylboronic acid β-CD-NH2 are dissolved in DMF and stirred at 60° C. for 1 h; γ-PGA and DCC are dissolved in DMF and stirred at 60° C. for 1 h.

[0053] The rest is the same as in Example 1, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0054] Example 10 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 1, specifically: Steps S1 and S2 are the same as those in Example 1 and will not be repeated here.

[0055] In step S3, DMAP and 3-aminophenylboronic acid β-CD-NH2 are dissolved in DMF, and stirred at 60° C. for 2 h; γ-PGA and DCC are dissolved in DMF, and stirred at 60° C. for 2 h.

[0056] The rest is the same as in Example 1, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0057] Embodiment 11 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 1, specifically: Steps S1 and S2 are the same as those in Example 1 and will not be repeated here.

[0058] In step S3, DMAP and 3-aminophenylboronic acid β-CD-NH2 are dissolved in DMF and stirred at 60° C. for 1 h; γ-PGA and DCC are dissolved in DMF and stirred at 60° C. for 1 h; after the reaction solutions are combined, the reaction is continued for 48 h.

[0059] The rest is the same as in Example 1, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0060] Example 12 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 1, specifically: Steps S1 and S2 are the same as those in Example 1 and will not be repeated here.

[0061] In step S3, DMAP and 3-aminophenylboronic acid β-CD-NH2 are dissolved in DMF and stirred at 60° C. for 2 h; γ-PGA and DCC are dissolved in DMF and stirred at 60° C. for 2 h; after the reaction solutions are combined, the reaction is continued for 48 h.

[0062] The rest is the same as in Example 1, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0063] Example 13 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material, and the specific preparation steps are as follows: Steps S1 and S2 are the same as those in Example 1 and will not be repeated here.

[0064] S3, weigh 5g DMAP, 1g 3-aminophenylboronic acid, 1.3g β-CD-NH2, dissolve in 100ml DMF, 500rt, 40℃ stirring for 1h; weigh 3.2g γ-PGA, 7g DCC, dissolve in 50ml DMF, 500rt, 40℃ stirring for 1h, combine the reaction solution and continue to react for 24h. After the reaction is completed, drop it into 1000ml ice water, precipitate, filter to obtain filtrate, concentrate the filtrate to 40ml, transfer the liquid and add 200ml EA to extract, obtain the lower layer liquid, continue to extract the lower layer liquid 2 times, drop the extracted lower layer solution into 200ml acetone to obtain the final product. The obtained product is the γ-PGA carrier material functionalized with 3-aminophenylboronic acid and β-CD-NH2.

[0065] Embodiment 14 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 13, specifically: Steps S1 and S2 are the same as those in Example 13 and will not be repeated here.

[0066] In step S3, DMAP and 3-aminophenylboronic acid β-CD-NH2 are dissolved in DMF, and stirred at 40° C. for 2 h; γ-PGA and DCC are dissolved in DMF, and stirred at 40° C. for 2 h.

[0067] The rest is the same as Example 13, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0068] Embodiment 15 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 13, specifically: Steps S1 and S2 are the same as those in Example 13 and will not be repeated here.

[0069] After the reaction solutions are combined in step S3, the reaction is continued for 48 hours.

[0070] The rest is the same as Example 13, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0071] Example 16 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 13, specifically: Steps S1 and S2 are the same as those in Example 13 and will not be repeated here.

[0072] In step S3, DMAP and 3-aminophenylboronic acid β-CD-NH2 are dissolved in DMF and stirred at 40° C. for 2 h; γ-PGA and DCC are dissolved in DMF and stirred at 40° C. for 2 h; after the reaction solutions are combined, the reaction is continued for 48 h.

[0073] The rest is the same as Example 13, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0074] Embodiment 17 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 13, specifically: Steps S1 and S2 are the same as those in Example 13 and will not be repeated here.

[0075] In step S3, DMAP and 3-aminophenylboronic acid β-CD-NH2 are dissolved in DMF, and stirred at 60° C. for 1 h; γ-PGA and DCC are dissolved in DMF, and stirred at 60° C. for 1 h.

[0076] The rest is the same as Example 13, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0077] Embodiment 18 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 13, specifically: Steps S1 and S2 are the same as those in Example 13 and will not be repeated here.

[0078] In step S3, DMAP and 3-aminophenylboronic acid β-CD-NH2 are dissolved in DMF, and stirred at 60° C. for 2 h; γ-PGA and DCC are dissolved in DMF, and stirred at 60° C. for 2 h.

[0079] The rest is the same as Example 13, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0080] Embodiment 19 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 13, specifically: Steps S1 and S2 are the same as those in Example 13 and will not be repeated here.

[0081] In step S3, DMAP and 3-aminophenylboronic acid β-CD-NH2 are dissolved in DMF and stirred at 60° C. for 1 h; γ-PGA and DCC are dissolved in DMF and stirred at 60° C. for 1 h; after the reaction solutions are combined, the reaction is continued for 48 h.

[0082] The rest is the same as Example 13, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0083] Embodiment 20 This embodiment provides a method for preparing a multi-functionalized γ-PGA carrier material. The specific preparation steps are similar to those in Example 13, specifically: Steps S1 and S2 are the same as those in Example 13 and will not be repeated here.

[0084] In step S3, DMAP and 3-aminophenylboronic acid β-CD-NH2 are dissolved in DMF and stirred at 60° C. for 2 h; γ-PGA and DCC are dissolved in DMF and stirred at 60° C. for 2 h; after the reaction solutions are combined, the reaction is continued for 48 h.

[0085] The rest is the same as Example 13, and the final product is 3-aminophenylboronic acid and β-CD-NH2 functionalized γ-PGA carrier material.

[0086] Figure 2-Figure 6 The SEM images of the γ-PGA carrier materials prepared in Examples 5, 8, 11, 13 and 16 loaded with the anti-tumor drug doxorubicin are shown respectively. It can be seen from the images that the γ-PGA molecular structure prepared in the present invention has multiple functional groups, a high drug loading rate and good physical and chemical properties.

[0087] Therefore, the present invention prepares a high molecular nano drug-carrying material with biodegradable γ-polyglutamic acid as the main chain, β-CD and 3-aminophenylboronic acid as the side chains, and utilizes the carboxyl group of γ-polyglutamic acid to load ions on the one hand, and on the other hand, the carboxyl group is also conducive to subsequent modification. The material degrades into glutamic acid in the body, which can achieve a therapeutic effect. The β-CD cavity can encapsulate ions, drug molecules, etc. Aminophenylboronic acid has fluorescence, so its fluorescence can be used to characterize the compound, and it has a boron hydroxyl group that can complex ions.

[0088] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that they can still modify or replace the technical solution of the present invention with equivalents, and these modifications or equivalent replacements cannot cause the modified technical solution to deviate from the spirit and scope of the technical solution of the present invention.

Claims

1. A multifunctionalized γ-PGA carrier material, characterized in that: The chemical structural formula of the multifunctionalized γ-PGA carrier material is: ； Among them, m is 150-350 and n is 6-8.

2. The method for preparing a multifunctionalized γ-PGA carrier material according to claim 1, characterized in that: The specific steps include: S1. Take β-cyclodextrin and p-toluenesulfonyl chloride, dissolve them in NaOH solution, stir to react, and post-treat to obtain compound C 49 H7O 37 S, chemical structure formula: ； S2, compound C obtained in step S1 49 H7O 37 S is added to ethylenediamine for reaction, and then post-processed to obtain compound C 48 H 84 N2O 34 , the chemical structure is: ； S3, compound C obtained in step S2 48 H 84 N2O 34 3-aminophenylboronic acid and 4-dimethylaminopyridine were placed in a reaction bottle and activated with DMF solvent. Meanwhile, γ-PGA and DCC were placed in another reaction bottle and activated with DMF solvent. The liquids in the two reaction bottles were mixed for reaction and post-processed to obtain the target product.

3. The method for preparing a multifunctionalized γ-PGA carrier material according to claim 2, characterized in that: In step S1, the molar mass ratio of β-cyclodextrin, p-toluenesulfonyl chloride and NaOH is one of 1:40:5, 1:45:7 or 1:50:10; the reaction temperature is 0° C., and the stirring reaction time is 60-120 min.

4. The method for preparing a multifunctionalized γ-PGA carrier material according to claim 2, characterized in that: In step S1, the post-treatment method is: filter the solution after the reaction, take the filtrate, adjust the pH to 2-5, stand for 8-24 hours to obtain a precipitate, filter and dry to obtain the amination product C 49 H7O 37 S.

5. The method for preparing a multifunctionalized γ-PGA carrier material according to claim 2, characterized in that: In step S2, C 49 H7O 37 The molar mass ratio of S to ethylenediamine is one of 1:50, 1:100, 1:180 or 1:200; the reaction temperature is 30-60° C., and the reaction time is 24-72 hours.

6. The method for preparing a multifunctionalized γ-PGA carrier material according to claim 2, characterized in that: In step S2, the post-treatment method is: pouring the reaction solution into a sufficient amount of acetone to obtain a precipitate, and filtering to obtain product C 48 H 84 N2O 34 .

7. The method for preparing a multi-functionalized γ-PGA carrier material according to claim 2, characterized in that: In step S3, 3-aminophenylboronic acid, C 48 H 84 N2O 34 The molar mass ratio of γ-PGA to DMAP is 6:1:50, 6:1:100 or 6:1:150; the mass ratio of γ-PGA to DCC is 1:1-4; the activation time is 60-120 minutes; the mixing reaction time is 24-72 hours, and the mixing reaction temperature is 40-80°C.

8. The method for preparing a multi-functionalized γ-PGA carrier material according to claim 2, characterized in that: In step S3, the post-treatment method is: drop the reaction solution into ice water, filter to obtain a filtrate, add ethyl acetate to the filtrate to extract until the upper liquid is clear, take the lower solution, concentrate the lower liquid and drop it into acetone to precipitate, and filter to obtain the target product.

9. The use of a multifunctionalized γ-PGA carrier material as claimed in claim 1, characterized in that: The multifunctionalized γ-PGA carrier material is used for carrying drug molecules.

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