AMINO ACID-BASED GLASS, PREPARATION METHOD AND USE THEREOF

Abstract

The present invention discloses biodegradable glass based on an amino acid, a peptide and a derivative, as well as the preparation method and use thereof. The main raw material of the glass is one or more of an amino acid, a peptide, a derivative or salt thereof. Compared with traditional glass, the glass of the present invention has significant advantages such as high biocompatibility, biodegradability, being 3D printable, and being compostable, etc., and its preparation process is simple and green, which can effectively avoid the influence of the traditional glass on the ecological environment. The glass of the present invention has a wide range of applications in the fields such as medicine, building material, chemical industry, food, electronics, national defense, etc., including but not limited to tissue engineering, tooth/bone repair, drug sustained-release, cell/protein sequestration, optical fiber communication, coatings, precision instruments, etc.

Claims

1. Amino acid-based glass, wherein the main raw material of the glass is one or more of an amino acid represented by formula (1), a peptide, their derivatives, or salts thereof, and the content of the main raw material in the glass is 70 wt % or more, preferably 80 wt % or more, further preferably 90 wt % or more; ##STR00002## the amino acid is one or more selected from glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, selenocysteine, and pyrrolysine: the peptide is a molecule formed by condensation of n above-mentioned amino acids via peptide bonds, where n?2, preferably 2?n?10; the derivatives of the amino acid or peptide are amino acids or peptides having protecting group(s) on the amino group P1 and/or the carboxyl group P2, wherein: the protecting group on the amino group P1 is any one or more selected from Trt, Boc, Fmoc, Cbz/Z, Allyl, C.sub.2-C.sub.18acyl, benzoyl, and naphthoyl: the protecting group on the carboxyl group P2 is any one or more selected from OFm, Othu, OBzl, OAll, OMe, and OEt: the amino group P1 and the carboxyl group P2 are protected individually or simultaneously.

2. The amino acid-based glass according to claim 1, wherein the glass is prepared completely from the amino acid, peptide and their derivatives.

3. The amino acid-based glass according to claim 1, wherein the glass is prepared from a single molecule of the following: a single amino acid molecule, a single peptide molecule, a single amino acid derivative, or a single peptide derivative; or the glass is composed of a combination of two or more molecules including: a combination of amino acid molecules, a combination of peptide molecules, a combination of amino acid derivative molecules, a combination of peptide derivative molecules, a combination of amino acid molecules and peptide molecules, a combination of amino acid molecules and amino acid derivative molecules, a combination of amino acid molecules and peptide derivative molecules, a combination of peptide molecules and amino acid derivative molecules, a combination of peptide molecules and peptide derivative molecules, a combination of amino acid derivative molecules and peptide derivative molecules, a combination of amino acid molecules, peptide molecules and amino acid derivative molecules, a combination of amino acid molecules, peptide molecules and peptide derivative molecules, a combination of amino acid molecules, amino acid derivative molecules and peptide derivative molecules, and a combination of amino acid molecules, peptide molecules, amino acid derivative molecules and peptide derivative molecules.

4. The amino acid-based glass according to claim 1, wherein the glass further comprises auxiliary raw materials selected from one of or a mixture of two or more of clarifiers, fluxes, opacifiers, and colorants.

5. The amino acid-based glass according to claim 1, wherein the glass has a hardness of between 420?550 HV, preferably between 500?550 HV; and the glass has a transparency of 30% or more, preferably 60% or more, further preferably 80%-91%.

6. The amino acid-based glass according to claim 1, wherein the glass has a brittleness index (m) of between 10?100, preferably between 20?50.

7. A preparation method for the amino acid-based glass according to claim 1, comprising the steps of: raising the temperature of raw materials to a temperature higher than the melting point (T.sub.m) under an inert gas atmosphere, and maintaining at this temperature for a period of time, lowering the temperature to below room temperature, and transferring the temperature-lowered sample to an annealing furnace for annealing treatment.

8. The preparation method according to claim 7, wherein the temperature higher than the melting point is a temperature higher than the melting point by 5?200 K, preferably higher than the melting point by 10?50 K: and the time for maintaining is 5 min?1 h, preferably 15?30 min.

9. The preparation method according to claim 8, wherein the temperature for the annealing treatment is a temperature lower than the glass transition temperature (T.sub.g) by 20?100 K, preferably 20?50 K; and the time for the annealing treatment is 5 min?3 h, preferably 15 min?1 h.

10. Use of the amino acid-based glass according to claim 1, including the use for additive manufacturing, composting, tissue engineering, tooth or bone repair, drug sustained-release, cell or protein sequestration, optical fiber communication, coatings, and precision instruments.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0087] FIG. 1 shows the physical picture of the Ac-Lys glass prepared in Example 1 at room temperature, which can be processed into glass beads or glass coatings.

[0088] FIG. 2 shows the DSC-TGA diagrams of the Ac-Lys glass prepared in Example 1, of which the melting temperature T.sub.m=536.70 K, and the weight loss was not significant at the melting point temperature, indicating that Ac-Lys did not decompose when melted at a high temperature.

[0089] FIG. 3 shows the DSC diagram of the Ac-Lys glass prepared in Example 1, of which the glass transition temperature T.sub.g=295.10 K.

[0090] FIG. 4 shows the H-NMR spectrum of the Z-Phe-Phe glass prepared in Example 2. Compared to the Z-Phe-Phe raw material, the peaks did not change significantly, indicating that the chemical composition of the raw material peptide molecules did not change when subjected to heating, melting and annealing treatments.

[0091] FIG. 5 shows the light transmittance of the Z-Phe-Phe glass prepared in Example 2, which is comparable to commercially available glass.

[0092] FIG. 6 shows the DSC diagram of the Z-Phe-Phe glass prepared in Example 2, of which the glass transition temperature T.sub.g=320.75 K.

[0093] FIG. 7 shows the picture of Boc-Gly powders and Boc-Gly glass prepared in Example 3 under a polarizing microscope, demonstrating that the glass formed was amorphous.

[0094] FIG. 8 shows the test results for the biocompatibility of the Boc-Gly glass prepared in Example 3, in which the glass was processed into a square coating with a width of 2 cm, co-incubated with 3T3 cells, and the cell activity was tested by the MTT method.

[0095] FIG. 9 shows the test results for mechanical properties of the Boc-Ala glass prepared in Example 4.

[0096] FIG. 10 shows the biodegradation curve of the Boc-Ala glass prepared in Example 4 in a compost soil sample, in which the initial mass of the glass sample was 42.58 mg.

[0097] FIG. 11 shows the performance test results for the mixed glass prepared in Example 5.

[0098] FIG. 12 shows the degradation of the mixed glass prepared in Example 5 in an artificial gastric juice (following the preparation method of the Chinese Pharmacopoeia).

[0099] FIG. 13 shows the body weight change of mice after gastric perfusion of the mixed glass prepared in Example 5. The cycle of gastric perfusion to mice is once every 5 days with a dose of 5 mg kg.sup.?1 for an observation period of 30 days, resulting in the number of gastric perfusions of 5.

[0100] FIG. 14 shows the pattern printed with a 3D printing equipment from the mixed glass prepared in Example 6, in which the mixed powders were placed in the barrel of the 3D printing equipment, and the heating temperature was set to 450 K.

[0101] FIG. 15 shows the degradation of the mixed glass prepared in Example 6 after being implanted in an animal mouse model.

[0102] FIG. 16 shows the biodegradation of the insulin-loaded amino acid-based glass prepared in Example 7 after being subcutaneously implanted in mice over time.

[0103] FIG. 17 shows the blood glucose change of diabetic mice after oral gastric perfusion of the insulin-loaded amino acid-based glass prepared in Example 7.

DETAILED DESCRIPTION

[0104] The technical solutions of the present invention will be described in detail below by way of examples, but the protection scope of the present invention is not limited thereto.

Example 1

[0105] A method for preparing a lysine-based glass comprises the steps of: [0106] (1) Weighing 20 mg of N-acetyl-L-lysine (Ac-Lys) powders, grinding in a mortar uniformly, then transferring to a crucible; [0107] (2) Placing the crucible containing Ac-Lys powders from step (1) in a heating device under N.sub.2 atmosphere; [0108] (3) Heating the crucible from room temperature to 600 K at a heating rate of 10 K min.sup.?1 by conducting a heating treatment on the device from step (2), and maintaining at this temperature for 10 min; [0109] (4) Lowering the temperature of the crucible to a temperature of 273.15 K at a cooling rate of 10 K min.sup.?1 by conducting a cooling treatment on the device from step (3); and [0110] (5) Transferring the sample from step (4) to an annealing furnace at a temperature of 283.15 K, and maintaining at this temperature for 20 min for conducting an annealing treatment on the glass, thereby obtaining Ac-Lys glass.

[0111] FIG. 1 shows the physical picture of the Ac-Lys glass prepared in Example 1 at room temperature, which can be processed into glass beads or glass coatings.

[0112] FIG. 2 shows the DSC-TGA diagrams of the Ac-Lys glass prepared in Example 1, of which the melting temperature T.sub.m=536.70 K, and the weight loss was not significant at the melting temperature, indicating that Ac-Lys did not decompose when melted at a high temperature.

[0113] FIG. 3 shows the DSC diagram of the Ac-Lys glass prepared in Example 1, of which the glass transition temperature T.sub.g=295.10 K.

Example 2

[0114] A method for preparing phenylalanine-based peptide glass comprises the steps of: [0115] (1) Weighing 50 mg of benzyloxycarbonyl-phenylalanyl-phenylalanyl (Z-Phe-Phe) powders, grinding in a mortar uniformly, then transferring to a crucible; [0116] (2) Placing the crucible containing Z-Phe-Phe powders from step (1) in a heating device under N.sub.2 atmosphere; [0117] (3) Heating the crucible from room temperature to 500 K at a heating rate of 40 K min.sup.?1 by conducting a heating treatment on the device from step (2), and maintaining at this temperature for 20 min; [0118] (4) Lowering the temperature of the crucible to a temperature of 273.15 K at a cooling rate of 50 K min.sup.?1 by conducting a cooling treatment on the device from step (3); and [0119] (5) Transferring the sample from step (4) to an annealing furnace at a temperature of 283.15 K, and maintaining at this temperature for 10 min for conducting an annealing treatment on the glass, thereby obtaining Z-Phe-Phe glass.

[0120] FIG. 4 shows the H-NMR spectrum of the Z-Phe-Phe glass prepared in Example 2. Compared to the Z-Phe-Phe raw material, the peaks did not change significantly, indicating that the chemical composition of the raw material peptide molecules did not change when subjected to heating, melting and annealing treatments.

[0121] FIG. 5 shows the light transmittance of the Z-Phe-Phe glass prepared in Example 2, which is comparable to commercially available glass.

[0122] FIG. 6 shows the DSC diagram of the Z-Phe-Phe glass prepared in Example 2, of which the glass transition temperature T.sub.g=320.75 K.

Example 3

[0123] A method for preparing glycine-based glass comprises the steps of: [0124] (1) Weighing 30 mg of N-tert-butoxycarbonyl-L-glycine (Boc-Gly) powders, grinding in a mortar uniformly, then transferring to a crucible; [0125] (2) Placing the crucible containing Boc-Gly powders from step (1) in a heating device under N.sub.2 atmosphere; [0126] (3) Heating the crucible from room temperature to 600 K at a heating rate of 10 K min.sup.?1 by conducting a heating treatment on the device from step (2), and maintaining at this temperature for 30 min; [0127] (4) Lowering the temperature of the crucible to a temperature of 273.15 K at a cooling rate of 10 K min by conducting a cooling treatment on the device from step (3); and [0128] (5) Transferring the sample from step (4) to an annealing furnace at a temperature of 283.15 K, and maintaining at this temperature for 30 min for conducting an annealing treatment on the glass, thereby obtaining Boc-Gly glass.

[0129] FIG. 7 shows the picture of Boc-Gly powders and Boc-Gly glass prepared in Example 3 under a polarizing microscope, demonstrating that the glass formed was amorphous.

[0130] FIG. 8 shows the test results for the biocompatibility of the Boc-Gly glass prepared in Example 3, in which the glass was processed into a square coating with a width of 2 cm, co-incubated with 3T3 cells, and the cell activity was tested by the MTT method. It is noted that the glass prepared in Example 3 did not dissolve in a neutral aqueous solution.

Example 4

[0131] A method for preparing alanine-based glass comprises the steps of: [0132] (1) Weighing 20 mg of N-tert-butoxycarbonyl-L-alanine (Boc-Ala) powders, grinding in a mortar uniformly, then transferring to a crucible; [0133] (2) Placing the crucible containing Boc-Ala powders from step (1) in a heating device under N.sub.2 atmosphere; [0134] (3) Heating the crucible from room temperature to 650 K at a heating rate of 5 K by conducting a heating treatment on the device from step (2), and maintaining at this temperature for 5 min; [0135] (4) Lowering the temperature of the crucible to a temperature of 273.15 K at a cooling rate of 20 K min by conducting a cooling treatment on the device from step (3); and [0136] (5) Transferring the sample from step (4) to an annealing furnace at a temperature of 283.15 K, and maintaining at this temperature for 10 min for conducting an annealing treatment on the glass, thereby obtaining Boc-Ala glass.

[0137] FIG. 9 shows the test results for mechanical properties of the Boc-Ala glass prepared in Example 4.

[0138] FIG. 10 shows the biodegradation curve of the Boc-Ala glass prepared in Example 4 in a compost soil sample, in which the initial mass of the glass sample was 42.58 mg.

Example 5

[0139] A method for preparing phenylalanine and glutamic acid-based glass comprises the steps of: [0140] (1) Weighing 10 mg of L-phenylalanine ethyl ester (Phe-OEt) powders and 10 mg of N-tert-butoxycarbonyl-L-glutamic acid dimethyl ester (Boc-Glu-dME) powders, grinding the mixture in a mortar uniformly, thereto adding 0.1 wt % of copper sulfate powders, and then grinding uniformly before transferring to a crucible; [0141] (2) Placing the crucible containing the mixed amino acids from step (1) in a heating device under N.sub.2 atmosphere; [0142] (3) Heating the crucible from room temperature to 550 K at a heating rate of 10 K min by conducting a heating treatment on the device from step (2), and maintaining at this temperature for 10 min; [0143] (4) Lowering the temperature of the crucible to a temperature of 273.15 K at a cooling rate of 10 K min by conducting a cooling treatment on the device from step (3); and [0144] (5) Transferring the sample from step (4) to an annealing furnace at a temperature of 283.15 K, and maintaining at this temperature for 10 min for conducting an annealing treatment on the glass, thereby obtaining Phe-OEt/Boc-Glu-dME mixed glass.

[0145] FIG. 11 shows the performance test results for the mixed glass prepared in Example 5.

[0146] FIG. 12 shows the degradation of the mixed glass prepared in Example 5 in an artificial gastric juice (following the preparation method of the Chinese Pharmacopoeia).

[0147] FIG. 13 shows the body weight change of mice after gastric perfusion of the mixed glass prepared in Example 5. The cycle of gastric perfusion to mice is once every 5 days with a dose of 5 mg kg for an observation period of 30 days, resulting in the number of gastric perfusions of 5.

Example 6

[0148] A method for preparing active peptide and amino acid derivative-based glass comprises the following steps: [0149] (1) Weighing 10 mg of immunoactive peptide Val-Gln-Pro-Ile-Pro-Tyr powders and 10 mg of N-tert-butoxycarbonyl-L-arginine methyl ester (Boc-L-Arg-OMe) powders, grinding the mixture in a mortar uniformly, then transferring to a crucible; [0150] (2) Placing the crucible containing the mixed powders from step (1) in a heating device under N.sub.2 atmosphere; [0151] (3) Heating the crucible from room temperature to 450 K at a heating rate of 10 K min.sup.?1 by conducting a heating treatment on the device from step (2), and maintaining at this temperature for 20 min; [0152] (4) Lowering the temperature of the crucible to a temperature of 273.15 K at a cooling rate of 10 K min.sup.?1 by conducting a cooling treatment on the device from step (3); and [0153] (5) Transferring the sample from step (4) to an annealing furnace at a temperature of 283.15 K, and maintaining at this temperature for 10 min for conducting an annealing treatment on the glass, thereby obtaining mixed glass.

[0154] FIG. 14 shows the pattern printed with a 3D printing equipment from the mixed glass prepared in Example 6, in which the mixed powders were placed in the barrel of the 3D printing equipment, and the heating temperature was set to 450 K.

[0155] FIG. 15 shows the degradation of the mixed glass prepared in Example 6 after being implanted in an animal mouse model.

Example 7

[0156] A method for preparing insulin-loaded amino acid-based glass comprises the following steps: [0157] (1) Weighing 50 mg of immunoactive peptide Val-Gln-Pro-Ile-Pro-Tyr powders, grinding in a mortar uniformly, then transferring to a crucible; [0158] (2) Placing the crucible containing the powders from step (1) in a heating device under N.sub.2 atmosphere; [0159] (3) Heating the crucible from room temperature to 450 K at a heating rate of 10 K min.sup.?1 by conducting a heating treatment on the device from step (2), and maintaining at this temperature for 10 min, then cooling to 330 K; [0160] (4) Weighing 5 mg of insulin powders, grinding in a mortar uniformly, then transferring to the crucible from step (3), stirring uniformly, and maintaining at this temperature for 10 min for melting; [0161] (5) Lowering the temperature of the crucible to a temperature of 273.15 K at a cooling rate of 20 K min.sup.?1 by conducting a cooling treatment on the device from step (4); and [0162] (6) Transferring the sample from step (5) to an annealing furnace at a temperature of 283.15 K, and maintaining at this temperature for 20 min for conducting an annealing treatment on the glass, thereby obtaining the insulin-loaded amino acid-based glass.

[0163] FIG. 16 shows the biodegradation of the insulin-loaded amino acid-based glass prepared in Example 7 after being subcutaneously implanted in mice over time.

[0164] FIG. 17 shows the blood glucose change of diabetic mice after oral gastric perfusion of the insulin-loaded amino acid-based glass prepared in Example 7.