Sericin protein particle with oxidative stress property, method for preparing the same and use thereof

Abstract

The application relates to a sericin protein particle with an oxidative stress property, a method for preparing the same and use thereof. The particle is formed by sericin protein and a tellurium compound. In the method for preparing the sericin protein particle, bis(1-hydroxydodecyl) telluride is first prepared and then mixed with a solution of sericin protein, and a resulting solution is agitated continuously for some time to obtain telluride-modified sericin protein. Then, the solution of the sericin protein is added dropwise into absolute ethanol to obtain a precipitated product. An aqueous solution of the precipitated product and a solution of magnesium ions are mixed and agitated for a certain time to obtain the sericin protein particle with an oxidative stress property. The sericin protein particle disclosed is a hollow particle having a diameter in a range of 600-1250 nm, and is capable of loading biomacromolecule drugs or other components.

Claims

1. A method for preparing sericin protein particles with an oxidative stress property, wherein the method comprises following steps: 1) adding sodium borohydride and tellurium powder into water, heating to a temperature suitable for a reaction under nitrogen protection in order to obtain a Na.sub.2Te.sub.2 aqueous solution; cooling the Na.sub.2Te.sub.2 aqueous solution to room temperature, adding 10-bromo-dodecyl alcohol and an organic solvent thereto, and heating to a temperature suitable for a reaction; and then performing post-treatment in order to obtain a white solid powder of bis(1-hydroxydodecyl) telluride; 2) preparing a telluride solution of the white solid powder of bis(1-hydroxydodecyl) telluride; 3) uniformly mixing the telluride solution prepared in step 2) with a solution of sericin protein, and adding a resulting solution dropwise into absolute ethanol for a reaction in order to obtain a white precipitate of telluride-modified sericin protein; and 4) preparing two solutions, wherein the two solutions are an aqueous solution of the telluride-modified sericin protein and an aqueous solution of magnesium ions; uniformly mixing the two solutions in order to obtain a mixed solution; adjusting the mixed solution with HCl and NaOH to have a pH value of 5.0-8.0 in order to obtain a pH-adjusted mixed solution; heating the pH-adjusted mixed solution to a temperature suitable for a reaction under a condition of agitating in order to obtain a resulting precipitate; and then separating the resulting precipitate in order to obtain sericin protein particles with an oxidative stress property.

2. The method for preparing a sericin protein particle with an oxidative stress property according to claim 1, wherein a molecular weight distribution of the sericin protein is 5-50 KDa.

3. The method for preparing a sericin protein particle with an oxidative stress property according to claim 1, wherein in step 4), after the aqueous solution of the telluride-modified sericin protein and the aqueous solution of the magnesium ions are mixed, a mass percentage concentration of the sericin protein is 0.05-2%, and a concentration of the magnesium ions is 5-15 mmol/L.

4. The method for preparing a sericin protein particle with an oxidative stress property according to claim 1, wherein the magnesium ions in step 4) are selected from any of magnesium chloride, magnesium nitrate, and magnesium sulfate.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is an SEM image of telluride-modified sericin particles prepared in Example 3;

(2) FIG. 2 is a TEM transmission slice image of the telluride-modified sericin particles prepared in Example 3;

(3) FIG. 3 is a DLS diagram showing particle diameter distribution of the telluride-modified sericin particles prepared in Example 3; and

(4) FIG. 4 is a DLS diagram showing particle diameter distribution of the telluride-modified sericin particles prepared in Example 3 after the telluride-modified sericin particles have been soaked in a 200 M of H.sub.2O.sub.2 solution for 24 hours.

DETAILED DESCRIPTION OF THE EMBODIMENTS

(5) The present disclosure will be further explained in conjunction with specific embodiments.

Example 1

(6) 1) 1.51 g of sodium borohydride and 4.84 g tellurium powder were added into 100 mL of water, and a resulting solution was heated to 80° C. under a condition of nitrogen protection to perform a reaction for 2.5 h to obtain a Na.sub.2Te.sub.2 aqueous solution. The Na.sub.2Te.sub.2 aqueous solution was cooled to room temperature, and 5.56 g of 10-bromo-dodecyl alcohol and 100 mL of tetrahydrofuran were added; and a resulting solution was heated to 50° C. to perform a reaction for 6 h. After the reaction was over, a resulting solution was cooled to room temperature, and a product was extracted by using dichloromethane. Then a dichloromethane phase was collected, and excess anhydrous magnesium sulfate was added for drying for 24 h; and a resulting solution was subjected to filtration and rotary evaporation to obtain a white solid powder of bis(1-hydroxydodecyl) telluride.

(7) 2) The obtained white solid powder of bis(1-hydroxydodecyl) telluride was dissolved in N,N-dimethylformamide to prepare a telluride solution having a mass concentration of 0.2%.

(8) 3) Sericin protein having a molecular weight of 5 KDa was dissolved in deionized water to prepare an aqueous solution of sericin protein with a mass concentration of 0.2%.

(9) 4) The telluride solution prepared in step 2) and the solution of sericin protein prepared in step 3) were mixed at a volume ratio of 0.2:1, and a resulting solution was agitated for 4 h at 40° C. at a revolving speed of 100 rpm. Then, the solution was added dropwise into absolute ethanol to obtain a white precipitate. The precipitate was separated by centrifugation, and the obtained precipitate was telluride-modified sericin protein.

(10) 5) An aqueous solution of the telluride-modified sericin protein and an aqueous solution of magnesium chloride were prepared respectively, and the two solutions were mixed uniformly to obtain a mixed solution, in which a mass percentage concentration of the sericin protein was 0.05%, and a concentration of magnesium ions was 5 mmol/L. After a system of the solution was adjusted with HCl and NaOH to have a pH value of 5.0, the solution was placed into a thermostat at a temperature of 30° C. to perform a reaction, during which the solution was agitated continuously for 2 h at a resolving speed of 300 rpm. Resulting precipitate was separated by centrifugation. The obtained precipitate was sericin protein particles with an oxidative stress property. An average diameter of the sericin protein particles was 1250 nm.

Example 2

(11) 1) 1.51 g of sodium borohydride and 4.84 g tellurium powder were added into 100 mL of water, and a resulting solution was heated to 80° C. under a condition of nitrogen protection to perform a reaction for 2.5 h to obtain a Na.sub.2Te.sub.2 aqueous solution. The Na.sub.2Te.sub.2 aqueous solution was cooled to room temperature, and 5.56 g of 10-bromo-dodecyl alcohol and 100 mL of tetrahydrofuran were added; and a resulting solution was heated to 50° C. to perform a reaction for 6 h. After the reaction was over, a resulting solution was cooled to room temperature, and a product was extracted by using dichloromethane. Then a dichloromethane phase was collected, and excess anhydrous magnesium sulfate was added for drying for 24 h; and a resulting solution was subjected to filtration and rotary evaporation to obtain a white solid powder of bis(1-hydroxydodecyl) telluride.

(12) 2) The obtained white solid powder of bis(1-hydroxydodecyl) telluride was dissolved in N,N-dimethylformamide to prepare a telluride solution having a mass concentration of 0.8%.

(13) 3) Sericin protein having a molecular weight of 10 KDa was dissolved in deionized water to prepare an aqueous solution of sericin protein with a mass concentration of 0.8%.

(14) 4) The telluride solution prepared in step 2) and the solution of sericin protein prepared in step 3) were mixed at a volume ratio of 0.4:1, and a resulting solution was agitated for 6 h at 40° C. at a revolving speed of 200 rpm. Then, the solution was added dropwise into absolute ethanol to obtain a white precipitate. The precipitate was separated by centrifugation, and the obtained precipitate was telluride-modified sericin protein.

(15) 5) An aqueous solution of the telluride-modified sericin protein and an aqueous solution of magnesium nitrate were prepared respectively, and the two solutions were mixed uniformly to obtain a mixed solution, in which a mass percentage concentration of the sericin protein was 0.2%, and a concentration of magnesium ions was 10 mmol/L. After a system of the solution was adjusted with HCl and NaOH to have a pH value of 6.0, the solution was placed into a thermostat at a temperature of 40° C. to perform a reaction, during which the solution was agitated continuously for 8 h at a resolving speed of 300 rpm. Resulting precipitate was separated by centrifugation. The obtained precipitate was sericin protein particles with an oxidative stress property. An average diameter of the sericin protein particles was 1080 nm.

Example 3

(16) 1) 1.51 g of sodium borohydride and 4.84 g tellurium powder were added into 100 mL of water, and a resulting solution was heated to 80° C. under a condition of nitrogen protection to perform a reaction for 2.5 h to obtain a Na.sub.2Te.sub.2 aqueous solution. The Na.sub.2Te.sub.2 aqueous solution was cooled to room temperature, and 5.56 g of 10-bromo-dodecyl alcohol and 100 mL of tetrahydrofuran were added; and a resulting solution was heated to 50° C. to perform a reaction for 6 h. After the reaction was over, a resulting solution was cooled to room temperature, and a product was extracted by using dichloromethane. Then a dichloromethane phase was collected, and excess anhydrous magnesium sulfate was added for drying for 24 h; and a resulting solution was subjected to filtration and rotary evaporation to obtain a white solid powder of bis(1-hydroxydodecyl) telluride.

(17) 2) The obtained white solid powder of bis(1-hydroxydodecyl) telluride was dissolved in N,N-dimethylformamide to prepare a telluride solution having a mass concentration of 1.5%.

(18) 3) Sericin protein having a molecular weight of 20 KDa was dissolved in deionized water to prepare an aqueous solution of sericin protein with a mass concentration of 1.5%.

(19) 4) The telluride solution prepared in step 2) and the solution of sericin protein prepared in step 3) were mixed at a volume ratio of 0.5:1, and a resulting solution was agitated for 6 h at 40° C. at a revolving speed of 200 rpm. Then, the solution was added dropwise into absolute ethanol to obtain a white precipitate. The precipitate was separated by centrifugation, and the obtained precipitate was telluride-modified sericin protein.

(20) 5) An aqueous solution of the telluride-modified sericin protein and an aqueous solution of magnesium sulfate were prepared respectively, and the two solutions were mixed uniformly to obtain a mixed solution, in which a mass percentage concentration of the sericin protein was 1%, and a concentration of magnesium ions was 10 mmol/L. After a system of the solution was adjusted with HCl and NaOH to have a pH value of 7.0, the solution was placed into a thermostat at a temperature of 40° C. to perform a reaction, during which the solution was agitated continuously for 10 h at a resolving speed of 300 rpm. Resulting precipitate was separated by centrifugation. The obtained precipitate was sericin protein particles C with an oxidative stress property. An average diameter of the sericin protein particles was 1000 nm.

(21) In addition, particle diameters of the telluride-modified sericin protein particles prepared in Example 3 were measured by dynamic light scattering. As shown in FIGS. 1-3, the particle diameters were in a range of 580-1580 nm, and an average particle diameter of the particles was 1000 nm. FIG. 4 shows a DLS diagram showing particle diameter distribution of the sericin protein particles after the sericin protein particles were soaked in 200 M of H.sub.2O.sub.2 solution for 24 h; and the particle diameters of the sericin protein particles increased to a range of 3000-6000 nm, and an average particle diameter thereof was 4000 nm. In the presence of H.sub.2O.sub.2, the sericin protein particles with an oxidative stress property underwent depolymerization, and the particle diameters thereof became larger.

Example 4

(22) 1) 1.51 g of sodium borohydride and 4.84 g tellurium powder were added into 100 mL of water, and a resulting solution was heated to 80° C. under a condition of nitrogen protection to perform a reaction for 2.5 h to obtain a Na.sub.2Te.sub.2 aqueous solution. The Na.sub.2Te.sub.2 aqueous solution was cooled to room temperature, and 5.56 g of 10-bromo-dodecyl alcohol and 100 mL of tetrahydrofuran were added; and a resulting solution was heated to 50° C. to perform a reaction for 6 h. After the reaction was over, a resulting solution was cooled to room temperature, and a product was extracted by using dichloromethane. Then a dichloromethane phase was collected, and excess anhydrous magnesium sulfate was added for drying for 24 h; and a resulting solution was subjected to filtration and rotary evaporation to obtain a white solid powder of bis(1-hydroxydodecyl) telluride.

(23) 2) The obtained white solid powder of bis(1-hydroxydodecyl) telluride was dissolved in N,N-dimethylformamide to prepare a telluride solution having a mass concentration of 2%.

(24) 3) Sericin protein having a molecular weight of 50 KDa was dissolved in deionized water to prepare an aqueous solution of sericin protein with a mass concentration of 2%.

(25) 4) The telluride solution prepared in step 2) and the solution of the sericin protein prepared in step 3) were mixed at a volume ratio of 1:1, and a resulting solution was agitated for 8 h at 40° C. at a revolving speed of 500 rpm. Then, the solution was added dropwise into absolute ethanol to obtain a white precipitate. The precipitate was separated by centrifugation, and the obtained precipitate was telluride-modified sericin protein.

(26) 5) An aqueous solution of the telluride-modified sericin protein and an aqueous solution of magnesium chloride were prepared respectively, and the two solutions were mixed uniformly to obtain a mixed solution, in which a mass percentage concentration of the sericin protein was 2%, and a concentration of magnesium ions was 15 mmol/L. After a system of the solution was adjusted with HCl and NaOH to have a pH value of 8.0, the solution was placed into a thermostat at a temperature of 50° C. to perform a reaction, during which the solution was agitated continuously for 20 h at a resolving speed of 300 rpm. Resulting precipitate was separated by centrifugation. The obtained precipitate was sericin protein particles with an oxidative stress property. An average diameter of the sericin protein particles was 600 nm.

(27) The above examples are only some embodiments of the present disclosure, and the protection scope of the present disclosure is not limited to the above examples; and the above examples do not represent all technical solutions under the inventive concept of the present disclosure. It shall be noted that, for those skilled in the art, additions and changes (for example, non-substantive modifications made to a concentration of sericin protein, a concentration of magnesium ions, a pH value, a temperature, a time length for a reaction and so on) may be made under the inspiration of the inventive concept of the present disclosure and specific examples without departing from the principles of the present disclosure, and these improvements and modifications shall also be considered as being within the protection scope of the present disclosure.