Tyrosine-derived amygdalin-loaded hydrogel preparation method and application

[0036] Example 1

[0037] Weigh out 5.0 mg/mL Fmoc-Tyr-OH solid powder, dissolve it in 100 μL dimethyl sulfoxide, disperse ultrasonically for 30 seconds to make it completely dissolved, and prepare a Fmoc-Tyr-OH stock solution with a concentration of 50 mg/mL; Take 5.0 mg of amygdalin in a centrifuge tube, add 1.0 mL of ultrapure water to dissolve it, and prepare a stock solution of amygdalin with a concentration of 5 mg/mL.

[0038] Take 50μL of Fmoc-Tyr-OH stock solution into a washed and dry 5.0mL screw-top bottle, add 970μL amygdalin stock solution, shake quickly, and let stand for 3 minutes to form a clear and transparent tyrosine derivative Hydrogel loaded with amygdalin.

[0039] Experimental steps of scanning electron microscopy:

[0040] Ultrasonic cleaning the silicon wafer with a water goby solution (concentrated sulfuric acid (v): hydrogen peroxide (v) = 7: 3) for 15 minutes, then ultrasonic cleaning with an ethanol solution for 15 minutes, and finally ultrapure water for 15 minutes, then remove the silicon wafer from Take out the piranha solution, rinse the silicon wafer with water twice, and then dry the silicon wafer with nitrogen. Use a pipette to take 10 μL of amygdalin hydrogel and place it on the silicon chip, put it in the refrigerator at -4°C, and freeze, and then put the frozen sample in a vacuum freeze dryer to remove the water. Put the prepared sample in a desiccator before SEM test. When performing SEM test, put the silicon wafer into the sample table, and then spray gold on the sample, put it into the instrument, and observe in a vacuum environment.

[0041] figure 1 Is a scanning electron microscope image of a tyrosine derivative supramolecular hydrogel, figure 2 Is a scanning electron microscope image of amygdalin solution, image 3 Load the tyrosine derivative with amygdalin supramolecular hydrogel scanning results. As shown in the figure, tyrosine derivatives can form a network structure, such as figure 1 As shown, tyrosine derivatives can be loaded with amygdalin, such as image 3 Shown. Figure 4 This is a digital photograph of a tyrosine derivative loaded amygdalin hydrogel prepared in Example 1 of the present invention

[0042] Example 2

[0043] Weigh out 5.0 mg/mL Fmoc-Tyr-OH solid powder, dissolve it in 100 μL dimethyl sulfoxide, disperse ultrasonically for 30 seconds to make it completely dissolved, and prepare a Fmoc-Tyr-OH stock solution with a concentration of 100 mg/mL; Take 16.0 mg of amygdalin in a centrifuge tube, add 1.0 mL of ultrapure water to dissolve it, and prepare a stock solution of amygdalin with a concentration of 16 mg/mL.

[0044] Take 30μL of Fmoc-Tyr-OH stock solution into a washed and dry 5.0mL screw-top bottle, add 950μL of amygdalin stock solution, shake quickly, and let stand for 3 minutes to form a clear and transparent tyrosine loaded bitter Hydrogel of amygdalin.

[0045] Fourier infrared test procedure: After drying the sample in a freeze dryer for 12 hours, weigh 0.2g of potassium bromide powder and then add 2mg of the sample, put it in a mortar and grind, mix the two thoroughly, and then The mixture is put into a mold and pressed into a sheet shape for infrared testing. Figure 5 Load the Fourier Infrared spectrum of amygdalin and amygdalin powder for tyrosine derivatives. The test result is like Figure 5 As shown, it can be obtained from the figure: In the infrared spectrum, the characteristic absorption bands of proteins and peptides are defined as amino acids A, B and I-VII. Amide Ⅰ (1600-1690cm -1 ) And amide Ⅱ (1480-1575cm -1 ) Is often used to analyze the secondary structure of proteins. In amide I, the absorption peaks of β-sheet, α-helix and random coil are respectively close to 1630 cm -1 , 1655cm -1 , 1645cm -1. Fmoc-Tyr-OH-Amygdalin which is at 1643cm -1 There is an absorption peak of amide Ⅰ, and Amygdalin (Amygdalin) is at 1642cm -1 There is an absorption peak. Amide II is derived from C-N stretching vibration and N-H bending. Both are at 1481cm -1 There are absorption peaks. These results indicate that the addition of amygdalin to Fmoc-Tyr-OH hydrogel did not cause a change in the structure of amygdalin.

[0046] Example 3

[0047] Weigh 5.0 mg/mL Fmoc-Tyr-OH solid powder, dissolve it in 100 μL of dimethyl sulfoxide, disperse ultrasonically for 60 seconds to make it completely dissolved, and prepare a Fmoc-Tyr-OH stock solution with a concentration of 75 mg/mL; Take 1.0 mg of amygdalin in a centrifuge tube, add 1.0 mL of ultrapure water to dissolve it, and prepare a stock solution of amygdalin with a concentration of 1 mg/mL.

[0048] Take 40μL of Fmoc-Tyr-OH stock solution into a washed and dried 5.0mL screw-top bottle, add 960μL amygdalin stock solution, shake quickly, and let stand for 3 minutes to form a clear and transparent tyrosine loaded bitter Amygdalin hydrogel.

[0049] Rheological test experiment procedure: use AR2000 rheological instrument to test its rheological behavior, use parallel plates with a diameter of 50mm, set the gap to 0.2mm, test conditions: temperature is 25℃, intensity is 2%, frequency is 0.1— 100Hz.

[0050] The test result is like Image 6 As shown, the storage modulus (G') loss modulus (G”) of Fmoc-Tyr-OH supramolecular hydrogel and Fmoc-Tyr-OH-Amygdalin supramolecular hydrogel were measured at the same time, where the storage modulus The quantity (G') represents the energy stored by the material due to elastic deformation, the loss modulus (G”) represents the energy lost due to the viscous deformation of the material, and the loss angle tangent (tanθ) is the loss modulus (G”) and storage The ratio of the modulus (G') is used to indicate the viscoelastic ratio of the material. In a general gel system, G'>G", that is, tanθ<1. Such as Image 6 As shown, in the frequency range of scanning, the value of G'is much greater than G", and after adding amygdalin to Fmoc-Tyr-OH supramolecular hydrogel, its G'is relatively simple Fmoc-Tyr-OH supramolecular hydrogel Glue increases. According to the above results, the loading of amygdalin in Fmoc-Tyr-OH supramolecular hydrogel will not affect its properties.