Preparation method of amphiphilic polyamino acid drug carrier

Abstract

The invention provides a preparation method of an amphiphilic polyamino acid drug carrier, which comprises the following steps: S1, dispersing chitosan in anhydrous dimethyl sulfoxide, heating to 40-60 DEG C, and stirring to fully dissolve the chitosan; s2, L-aspartic acid-beta-benzyl ester intracyclic anhydride is added, and a reaction is performed for 5-13 hours at the temperature of 20-40 DEG C; and S3, carrying out precipitation in water to remove dimethyl sulfoxide so as to obtain the chitosan-poly (L-aspartic acid-beta-benzyl ester) polymer. The polymer is completely prepared from CS, polyamino acid and other biological materials, and the carrier material has good biological safety and biodegradability. The polymer is simple in preparation method and low in raw material cost. Through molecular design and reaction condition control, the control of polymer structure, particle size and distribution can be realized, so that the influence of carrier structure and size effect on biological effect is known, and a scientific basis is provided for designing a nano-carrier with a reasonable structure.

Description

TECHNICAL FIELD

[0001] The present application relates to a preparation method of an amphiphilic polyamino acid drug carrier. BACKGROUND

[0002] Traditional anti-tumor drugs have the disadvantages of large toxic side effects and lack of specific recognition, which limits the application of chemotherapy drugs in tumor treatment. It is of great significance to improve the efficacy of tumor treatment by constructing drug carriers with good biological safety and intelligent responsiveness. Drug-loaded nanoparticles prepared by physical encapsulation, electrostatic adsorption, chemical bonding and other methods not only protect the activity of drugs, but also prolong the circulation time of drugs in the blood and improve drug utilization. Nanoparticles with a diameter of 100-200 nm can accumulate at the tumor site through enhanced permeability and retention effect, and respond to the tumor microenvironment to achieve targeted drug release.

[0003] Stimulus-responsive nanoparticles mainly include pH-responsive, redox-responsive, temperature-responsive, etc. The pH of normal tissues and body fluids is 7.4, while the pH of tumor extracellular is 6.8-7.2, the pH of endosomes in tumor cells is 5.5-6.0, and the pH of lysosomes is about 4.5. In addition, the concentration of glutathione (GSH) in tumor cytoplasm is 2-10 mmol / L, which is much higher than that in blood and normal cells, and the temperature of tumor tissue is higher than that of normal tissue. These characteristics of tumor microenvironment provide the basis for the construction of nanomedicine delivery system.

[0004] Although a large number of studies have been conducted on high molecular weight nanomedicine carriers, and a number of high molecular weight carrier materials have entered clinical application, there are still some problems in the research and application of high molecular weight nanomedicine carriers that need to be improved and solved.

[0005] First of all, good biological safety is a key problem for the practical application of high molecular weight nanomedicine carriers. In order to solve this problem, researchers select polymers with good biological safety, such as polyethylene glycol, polyethylene, and polycaprolactone, as the basic skeleton of high molecular weight carriers; by designing the structure of the carrier, controlling the polymerization method, and functionalizing modification, etc., to construct "intelligent" high molecular weight drug carriers. These studies have made good progress, but the in vivo biological safety and in vivo degradability of polymers still need to be further improved.

[0006] Secondly, the construction of a suitable structure of the carrier, the drug targeting delivery to the tumor lesion site, the realization of the drug responsive controlled release is the necessary requirement of the carrier. But for the current responsive polymer drug carrier, there are still the following problems need to be further researched and solved. Most of the responsive carriers have the shortcomings of single stimulation condition and low response sensitivity. Therefore, it is necessary to construct a drug carrier with multiple response to further improve the controlled release effect of the drug at a specific time and space, so as to achieve better tumor treatment effect.

[0007] Polyamino acid is one of the hotspots of functional material research, has good biocompatibility and biodegradability, through hydrolysis or enzymatic hydrolysis, poly(amino acid) can be degraded into small amino acids, can be absorbed by the human body or discharged outside the body. The existence of various characteristic functional groups such as amino or carboxyl in the structure makes poly(amino acid) material have potential application value in the field of pH response material. Hydrophilic and hydrophobic amino acid anhydride can be prepared into various ideal amphiphilic polymers by ring-opening polymerization, and can realize self-assembly in a specific solution, which is expected to be applied in the field of drug sustained release.

[0008] Chitosan (CS) is widely concerned in the biomedical field due to its excellent biodegradability, biocompatibility, mucus adhesion and antibacterial activity. Moreover, CS is relatively low in cost and can open the tight junction between cells, thereby increasing drug absorption. In this project, CS and polyamino acid will be used to design functional polymers with good biocompatibility, construct multiple-stimulus-responsive drug-loaded nanoparticles with targeting and drug release mechanism, improve the safety of drugs and their response sensitivity, and ultimately achieve the purpose of improving the treatment effect of cancer. The smooth implementation of this project will promote the research and development of the biological safety of antitumor drugs and the targeting and controlled release preparation technology, and improve the safety and treatment effect of antitumor drugs. SUMMARY

[0009] The application provides a preparation method of an amphiphilic polyamino acid drug carrier, which can effectively solve the above problems.

[0010] The application is implemented as follows:

[0011] The application provides a preparation method of an amphiphilic polyamino acid drug carrier, which comprises the following steps:

[0012] S1, dispersing chitosan in anhydrous dimethyl sulfoxide, heating to 40-60 DEG C, and stirring to fully dissolve;

[0013] S2, adding L-aspartic acid-β-benzyl ester cyclic anhydride, and reacting at 20-40 DEG C for 5-13 hours;

[0014] S3, precipitating in water to remove dimethyl sulfoxide, and obtaining chitosan-poly-L-aspartic acid-β-benzyl ester cyclic anhydride polymer.

[0015] As a further improvement, in step S1, the concentration of the anhydrous dimethyl sulfoxide is 0.1g-1g / 100ml.

[0016] As a further improvement, in step S2, the mass ratio of the chitosan to L-aspartic acid-β-benzyl ester cyclic anhydride is 1:2-10.

[0017] As a further improvement, in step S1, the heating to 40-60℃ and the stirring to fully dissolve include heating to 45-48℃ and stirring for 1 hour to fully dissolve.

[0018] As a further improvement, in step S3, the volume ratio of the water to dimethyl sulfoxide is 3-10:1

[0019] The present application has the following advantages: the polymer is prepared by using CS and polyamino acid and other biological materials, and the carrier material has good biological safety and biodegradability. In addition, the polymer preparation method is simple, and the raw material cost is low. Through molecular design and reaction condition control, the polymer structure, particle size and distribution can be controlled to understand the influence of the carrier structure and size effect on the biological effect, and to provide a scientific basis for designing a nano-carrier with a reasonable structure. DETAILED DESCRIPTION

[0020] To make the objectives, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described below in connection with the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the scope of protection of the present application. Therefore, the following detailed description of the embodiments of the present application is not intended to limit the scope of the claimed present application, but only represents selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the scope of protection of the present application.

[0021] Embodiment 1

[0022] A preparation method of an amphiphilic polyamino acid drug carrier, comprising the following steps: dispersing 0.5g of chitosan in 100ml of anhydrous dimethyl sulfoxide, heating to 46℃, and stirring for about 1 hour to fully dissolve; adding 2g of L-aspartic acid-β-benzyl ester cyclic anhydride, and reacting at 35℃ for 7 hours; then adding 500ml of water to precipitate and remove the dimethyl sulfoxide, to obtain a chitosan-poly-L-aspartic acid-β-benzyl ester cyclic anhydride polymer.

[0023] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. The present application can be variously changed and modified by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the scope of the present application.

Claims

1. A method for preparing an amphiphilic polyamino acid drug carrier, characterized in that, Includes the following steps: S1, disperse chitosan in anhydrous dimethyl sulfoxide, heat to 40-60°C, and stir to dissolve it completely; S2, add L-aspartic acid-β-benzyl ester ring anhydride, and react at 20-40℃ for 5-13 hours; S3, dimethyl sulfoxide is removed by precipitation in water to obtain chitosan-poly-L-aspartic acid-β-benzyl ester intracyclic acid polymer.

2. The method for preparing the amphiphilic polyamino acid drug carrier as described in claim 1, characterized in that, In step S1, the concentration of chitosan in anhydrous dimethyl sulfoxide is 0.1g to 1g / 100ml.

3. The method for preparing the amphiphilic polyamino acid drug carrier as described in claim 1, characterized in that, In step S2, the mass ratio of chitosan to L-aspartic acid-β-benzyl ester anhydride is 1:2 to 10.

4. The method for preparing the amphiphilic polyamino acid drug carrier as described in claim 1, characterized in that, In step S1, the specific steps of heating to 40-60°C and stirring to fully dissolve include: heating to 45-48°C and stirring for 1 hour to fully dissolve.

5. The method for preparing the amphiphilic polyamino acid drug carrier as described in claim 1, characterized in that, In step S3, the volume ratio of water to dimethyl sulfoxide is 3 to 10:1.