阅读说明：本技术 一种基于氨基酸的玻璃、其制备方法及应用 (Glass based on amino acid, preparation method and application thereof ) 是由 闫学海 邢蕊蕊 袁成前 于 2021-09-29 设计创作，主要内容包括：本发明公开了一种基于氨基酸、肽及衍生物的可生物降解玻璃,及其制备方法和应用,所述玻璃的主要原料为氨基酸、肽及其衍生物或其盐中的一种或者多种的组合。与传统的玻璃相比,本发明的玻璃具有高生物相容性、可生物降解、可3D打印、可堆肥等显著优势,其制备工艺简单、绿色,可有效避免传统玻璃对生态环境的影响；在医药、建材、化工、食品、电子、国防等领域具有广泛的应用,包括但不限制于组织工程、牙齿/骨骼修复、药物缓释、细胞/蛋白质封存、光纤通讯、涂层、精密仪器等。(The invention discloses biodegradable glass based on amino acid, peptide and derivatives, and a preparation method and application thereof. Compared with the traditional glass, the glass disclosed by the invention has the remarkable advantages of high biocompatibility, biodegradability, 3D printing, compostability and the like, the preparation process is simple and green, and the influence of the traditional glass on the ecological environment can be effectively avoided; the preparation method has wide application in the fields of medicine, building materials, chemical industry, food, electronics, national defense and the like, and comprises but is not limited to tissue engineering, tooth/bone repair, drug slow release, cell/protein encapsulation, optical fiber communication, coatings, precise instruments and the like.)

1. An amino acid-based glass, characterized in that the main raw material of the glass is one or a combination of more of amino acids represented by formula (1), peptides, derivatives thereof or salts thereof, and the content of the main raw material in the glass is more than 70 wt%, preferably more than 80 wt%, and more preferably more than 90 wt%;

the amino acid is selected from one or more of 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 refers to a molecule formed by condensing n amino acids through peptide bonds, wherein n is more than or equal to 2, preferably, n is more than or equal to 2 and less than or equal to 10;

the derivatives of the amino acids or peptides refer to: amino acids or peptides having protecting groups on the amino P1 and/or carboxyl P2, wherein:

the protecting group on the amino P1 is selected from Trt, Boc, Fmoc, Cbz/Z, Allyl, C₂-C₁₈Any one or more of acyl, benzoyl and naphthoyl;

the protecting group on the carboxyl P2 is selected from any one or combination of more of OFm, Otbu, 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 entirely from the amino acids, peptides and derivatives.

3. The amino acid based glass according to claim 1 or 2, wherein the glass is prepared from the following single molecules: a single amino acid molecule, a single peptide molecule, a single amino acid derivative, a single peptide derivative; alternatively, the first and second electrodes may be,

the glass is composed of a mixture of two or more molecules, the combination comprising:

amino acid molecule compositions, peptide molecule compositions, amino acid derivative molecule compositions, peptide molecule derivative compositions, amino acid molecule and peptide molecule compositions, amino acid and amino acid derivative compositions, amino acid and peptide derivative compositions, peptide and amino acid derivative molecule compositions, peptide and peptide derivative compositions, amino acid derivative and peptide derivative compositions, amino acid and peptide and amino acid derivative compositions, amino acid and peptide derivative compositions, amino acid and amino acid derivative and peptide derivative, amino acid and peptide derivative compositions.

4. The amino acid based glass according to claim 1, wherein the glass further comprises an auxiliary raw material selected from one or a mixture of two or more of a fining agent, a flux, an opacifying agent, and a coloring agent.

5. The amino acid based glass according to any of claims 1 to 4, wherein the glass hardness is between 420 and 550HV, preferably between 500 and 550 HV; the transparency of the glass is 30% or more, preferably 60% or more, and more preferably 80% to 91%.

6. The amino acid based glass according to any of claims 1 to 4, wherein the glass has a brittleness index (m) of between 10 and 100, preferably between 20 and 50.

7. A method for the production of amino acid based glass according to claims 1 to 6, characterized in that it comprises the following steps: heating the raw material to a temperature higher than the melting point (T) in an inert gas atmosphere_m) And carrying out heat preservation treatment for a period of time, then cooling to room temperature or below, and transferring the cooled 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 5-200K higher than the melting point, preferably 10-50K higher than the melting point; the heat preservation time is 5min to 1h, preferably 15min to 30 min.

9. The method of claim 8, wherein the annealing temperature is below the glass transition temperature (T ™)_g) The temperature is 20-100K, preferably lower than 20-50K; the annealing time is 5min to 3h, preferably 15min to 1 h.

10. Use of the amino acid based glass according to claims 1-6, comprising: it can be used for additive manufacturing, composting, tissue engineering, tooth or bone repair, drug release, cell or protein sealing, optical fiber communication, coating, and precision instrument.

The technical field is as follows:

the invention discloses a glass material, a preparation method and application thereof, and particularly relates to amino acid-based biomolecule glass, a preparation method and application thereof, belonging to the field of new materials.

Background art:

glass is generally produced from inorganic minerals such as silica and calcium carbonate as main raw materials, and is one of the most commonly used materials in daily life. Glass is hardly degradable under natural conditions and is easily broken, so that the influence of glass on environmental ecology is significant in terms of pollution, harmfulness and permanence.

Various glass materials, products and methods for producing the same have been disclosed, and for example, a method for producing a silicate glass, a silicate glass and a silica raw material for a silicate glass have been disclosed (WO2015/129495 JA 2015.09.03); a glass product using β -quartz or β -spodumene solid solution as a main raw material has been disclosed (WO2005/058766 EN 2005.06.30); lithium silicate glass ceramics and lithium silicate glasses comprising divalent metal oxides have been disclosed (WO2013/053864DE 2013.04.18).

It is worth mentioning that the bioglass invented in 1969 by the university of Florida L.L. Henschel has 45% Na as the main component₂O, 25% CaO and 25% SiO₂And 5% of P₂O₅Exemplary compositions and applications of bioglass (also referred to as bioactive glass) are disclosed (US 4478904A; US6338751B 1; US7569105B 2).

The glass materials and products disclosed above have in common that the raw materials are inorganic minerals. At present, a biomolecule glass material based on amino acid and a preparation method thereof are not disclosed.

Amino acids are the basic units constituting proteins, and peptides are compounds in which two or more amino acids are linked by peptide bonds. Amino acids and peptides are important components of living organisms, and play an extremely important role in information transmission, metabolism, diseases, aging, and the like of living organisms. The amino acid-based biomolecules have extremely high biocompatibility, and are biodegradable and have a definite metabolic mechanism in the living body. Surprisingly, it has been found that amino acids, peptides and derivatives thereof can be prepared into biodegradable glasses having a glassy structure at room temperature by a specific preparation process, and the present invention has been completed based on this finding. The amino acid-based biomolecule glass discovered based on the invention is expected to be a new material and is widely applied to the fields of medicine, building materials, chemical industry, food, electronics, national defense and the like.

The invention content is as follows:

the invention aims to provide amino acid-based biomolecule glass and a preparation method thereof, the glass is eco-friendly, has high biocompatibility, is biodegradable, can be printed in 3D and composted, and is simple and green in preparation process.

In a first aspect: the amino acid-based glass is characterized in that the main raw materials are amino acid, peptide and derivatives thereof represented by the formula (1), and the content of the main raw materials in the glass is more than 70 wt%, preferably more than 80 wt%, and more preferably more than 90 wt%.

The amino acids include: glycine, alanine, valine, leucine, isoleucine, methionine (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 condensing n amino acids through peptide bonds, wherein n is more than or equal to 2, preferably, n is more than or equal to 2 and less than or equal to 10.

The derivative is amino acid and peptide with amino (P1) and carboxyl (P2) protecting groups, wherein:

protecting groups at P1 include, but are not limited to, Trt, Boc, Fmoc, Cbz/Z, Allyl, C₂-C₁₈Acyl, benzoyl, naphthoyl; protecting groups at P2 include, but are not limited to, OFm, Otbu, OBzl, OAll, OMe, OEt;

protection at P1 and P2 individually or simultaneously.

The derivatives also include: molecules, isomers and salts thereof similar to the molecular structural skeleton of the amino acid molecules or peptide molecules or derivatives thereof.

In a second aspect, the amino acid-based glass is characterized by being prepared from all of the amino acids, peptides and derivatives.

In a third aspect, the amino acid-based glass described above is characterized in that:

the amino acid-based glass can be prepared for a single molecule comprising:

a single amino acid molecule, a single peptide molecule, a single amino acid derivative, a single peptide derivative;

can also be prepared by mixing two or more molecules, and comprises:

amino acid molecule compositions, peptide molecule compositions, amino acid derivative molecule compositions, peptide molecule derivative compositions, amino acid molecule and peptide molecule compositions, amino acid and amino acid derivative compositions, amino acid and peptide derivative compositions, peptide and amino acid derivative molecule compositions, peptide and peptide derivative compositions, amino acid derivative and peptide derivative compositions, amino acid and peptide and amino acid derivative compositions, amino acid and peptide derivative compositions, amino acid and amino acid derivative and peptide derivative, amino acid and peptide derivative compositions.

In a fourth aspect, the amino acid-based glass is characterized in that the amino acid-based glass can be added with auxiliary materials in addition to the main materials, and comprises: one or a mixture of more than two of clarifying agent, fluxing agent, opacifying agent and coloring agent;

wherein the proportion of the auxiliary raw materials is 0-5 wt%, preferably 0-1 wt%;

the clarifying agent comprises: one or more of antimony oxide, sodium nitrate, ammonium nitrate, sodium sulfate, calcium sulfate, sodium chloride and ammonium chloride;

the cosolvent is: one or more of sodium carbonate, potassium carbonate, sodium carbonate and potassium nitrate;

the opacifier is: one or a mixture of more than two of cryolite, sodium fluosilicate and tin phosphide;

the colorant is: metal compounds of transition elements such as cobalt, manganese, nickel, iron, and copper.

In a fifth aspect, a method for making an amino acid based glass comprises the steps of: heating the raw materials to a temperature higher than the melting point in an inert gas atmosphere, carrying out heat preservation treatment for a period of time, then cooling to room temperature or below, and transferring the cooled sample to an annealing furnace for annealing treatment.

In a preferred embodiment of the invention, said temperature above the melting point (T)_m) The temperature is 5-200K higher than the melting point temperature, preferably 10-50K higher than the melting point temperature; the heat preservation time is 5min to 1h, preferably 15min to 30 min.

In a preferred embodiment of the present invention, the annealing temperature is below the glass transition temperature (T)_g) The temperature is 20-100K, preferably lower than 20-50K; the annealing time is 5min to 3h, preferably 15min to 1 h.

In one embodiment of the present invention, the amino acid based glass is a single molecule glass, comprising the following preparation steps:

(1) weighing amino acid, peptide or derivative powder with a certain mass, placing the powder into a mortar, grinding the powder uniformly, and transferring the powder into a crucible;

(2) placing the crucible filled with the raw materials in the step (1) in heating equipment under an inert gas atmosphere;

(3) heating the equipment in the step (2) to S₁Heating the crucible from room temperature to M₁Temperature, holding treatment T at this temperature₁Time, wherein:

S₁1 to 50K min^-1Preferably, it is 2 to 10K min^-1；

M₁Is higher than T_mA temperature of 5 to 200K, preferably above T_m 10-50K；

T₁5min to 1h, preferably 15 to 30 min;

(4) cooling the equipment in the step (3) by S₂The cooling rate of (2) is to lower the crucible to M₂Temperature, wherein:

S₂1 to 100K min^-1Preferably 50 to 100K min^-1；

M₂273.15K (temperature of ice-water mixture) or 293.15-298.15K (room temperature/normal temperature);

(5) transferring the sample of step (4) to a temperature M₃In the annealing furnace, constant temperature T₃And (b) performing annealing treatment on the glass, wherein:

M₃is less than T_gA temperature of 20 to 100K, preferably less than T_g 20～50K；

T₃Is 5min to 3h, preferably 15min to 1 h.

For example, the glass mixed by two or more molecules comprises the following improvement steps:

(1 °) weighing one component of powder, respectively placing the powder in a mortar, uniformly grinding, and transferring to different crucibles;

(2 °) following step (2-3);

(3 ℃) the melted components are mixed in the same crucible according to the proportion, and are stirred properly to be uniform, the mixing proportion is preferably 1:1: … …;

(4 ℃) the mixture obtained in step (3 ℃) is stirred at M₁Heat preservation treatment at temperature T_sTime;

(5 °) following step (4-5);

or the steps are as follows:

(6 ℃) weighing each component powder respectively, and uniformly stirring and mixing according to the proportion, wherein the mixing proportion is preferably 1:1: … …;

(7 ℃) putting the uniformly mixed powder into a mortar for grinding uniformly, and transferring the powder into a crucible;

(8 °) the procedure (2-5) was followed.

According to a sixth aspect, a method for preparing amino acid-based glass is characterized in that, for example, auxiliary raw materials are added in addition to main raw materials, and the steps are as follows:

(1) weighing the main raw materials and the auxiliary raw materials, mixing and stirring uniformly according to a proportion, and transferring into a crucible;

(2) the step (2-5) of the fifth aspect is followed.

In a seventh aspect, said T_mAnd T_gMeasured by standard Differential Scanning Calorimetry (DSC) methods:

the temperature rise rate of the DSC is set, preferably 10K min^-1Taking the temperature as an abscissa and the heat flow as an ordinate, making a curve, recording the initial temperature and the end temperature of the melting of the sample, and taking the midpoint temperature of the initial temperature and the end temperature as T_m；

When the temperature rises to be higher than T_mAfter the temperature is 20K, preserving the heat for 10 min;

setting the temperature reduction rate of DSC, preferably 10K min^-1Reducing the temperature to 273.15K, and then preserving the heat for 10 min;

heating for the 2 nd time, setting the temperature rise rate of DSC, preferably 10K min^-1Taking the temperature as an abscissa and the heat flow as an ordinate, making a curve, recording the initial temperature and the end temperature of the glass transition by extrapolating a tangent, and taking the midpoint temperature of the initial temperature and the end temperature as T_g。

An amino acid-based glass prepared by the method.

In an eighth aspect, the amino acid-based glass and the preparation method thereof of the present invention have the following advantages and beneficial effects:

(1) the glass based on amino acid has the properties of hard texture, brittleness, transparency, light transmission and the like: the hardness is between 420 and 550HV, preferably between 500 and 550 HV; the transparency is distributed between 30% and 91%, preferably between 80% and 91%;

(2) the glass based on amino acid has better Glass Forming Ability (GFA), and the brittleness index (m) of the glass based on amino acid is 10-100, preferably 10-50;

(3) the glass based on amino acid is environment-friendly, has higher biocompatibility and can be biodegraded;

(4) the glass based on amino acid has simple preparation process, high repeatability, greenness and environmental protection;

(5) the amino acid based glass of the present invention can be used for additive manufacturing (3D printing);

(6) the amino acid-based glass can be used for composting, and the damage of the traditional glass to the ecological environment is greatly reduced.

In a ninth aspect, the amino acid based glass of the present invention has the following applications: the method can be applied to the fields of medicine, building materials, chemical industry, food, electronics, national defense and the like, and comprises but is not limited to the aspects of tissue engineering, tooth/bone repair, drug slow release, cell/protein encapsulation, optical fiber communication, coatings, precise instruments and the like.

In a tenth aspect, the amino acid-based glass of the present invention can melt a drug molecule during melting, preferably, the drug molecule is a drug molecule having a short half-life and/or a poorly soluble drug molecule;

the drug molecules comprise any one or a mixture of more than two of tumor chemotherapy drug molecules, contrast agent molecules, antipyretic analgesic anti-inflammatory molecules, traditional Chinese medicine monomers, immunomodulators and the like;

chemotherapeutic drug molecules include: one or more of pemetrexed, fluorouracil, adriamycin, paclitaxel, docetaxel, vincristine, cisplatin, tamoxifen, megestrol, goserelin and analogues thereof;

the contrast agent molecules include: barium sulfate, iodine preparation (sodium iodide, diatrizoate, iothalamate, iodixanoic acid, iohexol, iopromide, idole, iotrolan, iodized oil, and iodophenyl ester),¹⁸Any one or a mixture of more than two of FDG, Gd-DTPA, Mn-DPDP, SPIO and analogues thereof;

the antipyretic analgesic anti-inflammatory molecules include: one or more of aspirin, ibuprofen, acetaminophen, indomethacin, nimesulide, rofecoxib, celecoxib and analogues thereof;

the traditional Chinese medicine monomer molecules comprise: one or more of curcumin, nobiletin, methyl tripterygium wilfordii, radix astragali, coriolus versicolor polysaccharide and analogues thereof;

the immunomodulator comprises: any one or a mixture of more than two of glycoprotein, pidotimod, thymosin alpha 1, muramyl dipeptide, interferon gamma, interleukin-2, levamisole and analogues thereof;

the other is any one or the mixture of more than two of insulin, paliperidone, nifedipine, ranitidine hydrochloride and the like which need slow release.

The sustained release preparation can be used as a subcutaneous embedding agent, an oral agent and a tissue engineering scaffold material, preferably, the raw material is amino acid, peptide or a derivative thereof with bioactivity, and the sustained release preparation is characterized in that the sustained release preparation realizes local and sustained release of the medicament along with the biodegradation of glass based on the amino acid.

In an eleventh aspect, the amino acid-based glass of the present invention can be dissolved with other functional agents during the melting process or coated on the surface of glass material in the form of a coating for performing certain functions, including but not limited to electrical conductivity, sterilization/corrosion protection, and radiation protection;

the conductive agent includes: any one or a mixture of more than two of indium tin oxide, graphite, polyacetylene and the like;

the bactericidal/preservative includes: any one or a mixture of more than two of nano silver, chlorine preparation, peroxide, organic sulfur, organic bromine, nitrogen-sulfur heterocyclic compound and analogues thereof;

the radiation protective agent comprises any one or a mixture of more than two of melanin, polyimide and analogues thereof.

In a twelfth aspect, the method for melting a drug or functional agent in a melting process of the amino acid-based glass of the present invention may employ powder co-melting; also can adopt a preparation method of pre-dissolving the medicine or the functional preparation in a good solvent, blending with molten glass based on amino acid, and then removing the solvent, which is characterized in that:

the content of the drug molecules is 0.01-25 wt%, preferably 0.1-1 wt%;

the content of the functional molecules is 0.01-5 wt%, preferably 0.1-1 wt%.

Drawings

FIG. 1 is a schematic representation of the Ac-Lys glass prepared in example 1 at room temperature, and can be processed into glass beads or coated with glass.

FIG. 2 is a DSC-TGA chart of Ac-Lys glass prepared in example 1, having a melting temperature T_mWhen the melting point temperature was 536.70K, weight loss was not significant, indicating that Ac-Lys was not decomposed when melted at high temperature.

FIG. 3 is a DSC of Ac-Lys glass prepared in example 1, having a glass transition temperature T_g＝295.10K。

FIG. 4 is a nuclear magnetic hydrogen spectrum of the Z-Phe-Phe glass prepared in example 2, in which the peaks are not significantly changed compared to the Z-Phe-Phe starting material, indicating that the chemical composition of the peptide starting material molecules is not changed by heating, melting and annealing.

FIG. 5 shows the optical transmission of the Z-Phe-Phe glass prepared in example 2, which is comparable to commercially available glass.

FIG. 6 is a DSC spectrum of Z-Phe-Phe glass having a glass transition temperature T as prepared in example 2_g＝320.75K。

FIG. 7 is a photograph under a polarizing microscope of Boc-Gly powder and Boc-Gly glass prepared in example 3, demonstrating that the formed glass is amorphous.

FIG. 8 shows the results of biocompatibility test of Boc-Gly glass prepared in example 3, which was processed into a square coating of 2cm width, 3T3 cells were incubated therewith, and the activity of the cells was tested by MTT method.

FIG. 9 shows the results of mechanical property measurements of Boc-Ala glass prepared in example 4.

FIG. 10 is the biodegradation profile of the Boc-Ala glass prepared in example 4 in a compost soil sample, the initial mass of the glass sample being 42.58 mg.

FIG. 11 shows the results of performance tests on the hybrid glasses prepared in real-time example 5.

Fig. 12 shows the degradation of the mixed glass prepared in example 5 in artificial gastric juice (following the chinese pharmacopoeia preparation method).

FIG. 13 is a graph showing the change in body weight of mice after the mixed glass prepared in example 5 was subjected to gastric lavage. The gastric lavage period of the mouse is once every 5 days, and the mass is 5mgkg^-1The observation period is 30 days, and the times of intragastric administration are 5 times.

Fig. 14 shows a pattern printed by the mixed glass prepared in example 6 through a 3D printing device. The mixed powder was placed in a barrel of a 3D printer, and the heating temperature was set at 450K.

FIG. 15 shows the degradation of the mixed glass prepared in example 6 after implantation in an animal mouse model.

FIG. 16 is a graph of the biodegradation of insulin-loaded amino acid-based glass prepared in example 7, implanted subcutaneously in mice over time.

FIG. 17 is a graph showing the blood glucose changes in insulin-loaded amino acid-based glass prepared in example 7 after oral gavage in diabetic mice.

Detailed Description

The technical solution of the present invention will be described in detail by examples, but the present invention is not limited thereto.

Example 1

A preparation method of lysine-based glass comprises the following steps:

(1) weighing 20mg of N-acetyl-L-lysine (Ac-Lys) powder, placing the powder into a mortar, uniformly grinding the powder, and transferring the powder into a crucible;

(2) placing the crucible filled with Ac-Lys powder in the step (1) in N₂Placing the mixture in heating equipment under the atmosphere;

(3) heating the equipment in the step (2) for 10K min^-1The temperature of the crucible is increased from room temperature to 600K at the temperature increasing rate, and the crucible is subjected to heat preservation treatment at the temperature for 10 min;

(4) cooling the equipment in the step (3) for 10K min^-1The crucible is cooled to 273.15K at the cooling rate;

(5) and (5) transferring the sample in the step (4) to an annealing furnace with the temperature of 283.15K, keeping the temperature for 20min, and carrying out annealing treatment on the glass to obtain the Ac-Lys glass.

FIG. 1 is a schematic representation of the Ac-Lys glass prepared in example 1 at room temperature, and can be processed into glass beads or coated with glass.

FIG. 2 is a DSC-TGA chart of Ac-Lys glass prepared in example 1, having a melting temperature T_mWhen the melting temperature was 536.70K, weight loss was not significant, indicating that Ac-Lys was not decomposed when melted at high temperature.

FIG. 3 is a DSC of Ac-Lys glass prepared in example 1, having a glass transition temperature T_g＝295.10K。

Example 2

The preparation method of the phenylalanine-based peptide glass comprises the following steps:

50mg of benzyloxycarbonyl-phenylalanyl (Z-Phe-Phe) powder was weighed, put in a mortar and ground uniformly, and then transferred to a crucible;

(2) placing the crucible filled with the Z-Phe-Phe powder in the step (1) in N₂Placing the mixture in heating equipment under the atmosphere;

(3) heating the equipment in the step (2) for 40K min^-1The temperature of the crucible is increased from room temperature to 500K at the temperature increasing rate, and the crucible is subjected to heat preservation treatment at the temperature for 20 min;

(4) cooling the equipment in the step (3) for 50K min^-1The crucible is cooled to 273.15K at the cooling rate;

(5) and (5) transferring the sample in the step (4) to an annealing furnace with the temperature of 283.15K, keeping the temperature for 10min, and carrying out annealing treatment on the glass to obtain the Z-Phe-Phe glass.

FIG. 4 is a nuclear magnetic hydrogen spectrum of the Z-Phe-Phe glass prepared in example 2, in which the peaks are not significantly changed compared to the Z-Phe-Phe starting material, indicating that the chemical composition of the peptide starting material molecules is not changed by heating, melting and annealing.

FIG. 5 shows the optical transmission of the Z-Phe-Phe glass prepared in example 2, which is comparable to commercially available glass.

FIG. 6 is a DSC spectrum of Z-Phe-Phe glass having a glass transition temperature T as prepared in example 2_g＝320.75K。

Example 3

The preparation method of the glycine-based glass comprises the following steps:

(1) weighing 30mg of N-tert-butyloxycarbonyl-L-glycine (Boc-Gly) powder, placing the powder into a mortar, uniformly grinding the powder, and transferring the powder into a crucible;

(2) placing the crucible filled with Boc-Gly in the step (1) in N₂Placing the mixture in heating equipment under the atmosphere;

(3) heating the equipment in the step (2) for 10K min^-1The temperature of the crucible is increased from room temperature to 600K at the temperature increasing rate, and the crucible is subjected to heat preservation treatment at the temperature for 30 min;

(4) cooling the equipment in the step (3) for 10K min^-1The crucible is cooled to 273.15K at the cooling rate;

(5) and (5) transferring the sample in the step (4) to an annealing furnace with the temperature of 283.15K, keeping the temperature for 30min, and carrying out annealing treatment on the glass to obtain the Boc-Gly glass.

FIG. 7 is a photograph under a polarizing microscope of Boc-Gly powder and Boc-Gly glass prepared in example 3, demonstrating that the formed glass is amorphous.

FIG. 8 shows the results of biocompatibility test of Boc-Gly glass prepared in example 3, which was processed into a square coating of 2cm width, 3T3 cells were incubated therewith, and the activity of the cells was tested by MTT method. The glass prepared in example 3 was not dissolved in a neutral aqueous solution.

Example 4

The preparation method of the glass based on the alanine comprises the following steps:

(1) weighing 20mg of N-tert-butyloxycarbonyl-L-alanine (Boc-Ala) powder, placing the powder in a mortar, uniformly grinding the powder, and transferring the powder into a crucible;

(2) placing the crucible filled with Boc-Ala powder in step (1) in N₂Placing the mixture in heating equipment under the atmosphere;

(3) heating the equipment in the step (2) for 5K min^-1The temperature of the crucible is increased from room temperature to 650K at the temperature increasing rate, and the crucible is subjected to heat preservation treatment at the temperature for 5 min;

(4) to step (3)Cooling the equipment for 20K min^-1The crucible is cooled to 273.15K at the cooling rate;

(5) and (5) transferring the sample in the step (4) to an annealing furnace with the temperature of 283.15K, keeping the temperature for 10min, and carrying out annealing treatment on glass to obtain the Boc-Ala glass.

FIG. 9 shows the results of mechanical property measurements of Boc-Ala glass prepared in example 4.

FIG. 10 is the biodegradation profile of the Boc-Ala glass prepared in example 4 in a compost soil sample, the initial mass of the glass sample being 42.58 mg.

Example 5

The preparation method of the glass based on the phenylalanine and the glutamic acid comprises the following steps:

(1) weighing 10mg of L-phenylalanine ethyl ester powder (Phe-OEt) and 10mg of N-tert-butoxycarbonyl-L-glutamic acid dimethyl ester (Boc-Glu-dME) powder, uniformly grinding in a mortar, adding 0.1 wt% of copper sulfate powder, uniformly grinding, and transferring to a crucible;

(2) putting the crucible filled with the mixed amino acid in the step (1) in N₂Placing the mixture in heating equipment under the atmosphere;

(3) heating the equipment in the step (2) for 10K min^-1The temperature of the crucible is increased from room temperature to 550K at the temperature increasing rate, and the crucible is subjected to heat preservation treatment at the temperature for 10 min;

(4) cooling the equipment in the step (3) for 10K min^-1The crucible is cooled to 273.15K at the cooling rate;

(5) and (5) transferring the sample in the step (4) to an annealing furnace with the temperature of 283.15K, keeping the temperature for 10min, and carrying out annealing treatment on the glass to obtain the mixed glass of Phe-OEt/Boc-Glu-dME.

FIG. 11 shows the results of the performance test of the hybrid glass prepared in example 5.

Fig. 12 shows the degradation of the mixed glass prepared in example 5 in artificial gastric juice (following the chinese pharmacopoeia preparation method).

FIG. 13 is a graph showing the change in body weight of mice after the mixed glass prepared in example 5 was subjected to gastric lavage. The gavage period of the mice is 5 days to oneSecondly, the mass is 5mgkg^-1The observation period is 30 days, and the times of intragastric administration are 5 times.

Example 6

A preparation method of glass based on active peptide and amino acid derivative comprises the following steps:

(1) weighing 10mg of immune active peptide Val-Gln-Pro-Ile-Pro-Tyr powder and 10mg of N-tert-butyloxycarbonyl-L-arginine methyl ester (Boc-L-Arg-OMe) powder, uniformly grinding in a mortar, and transferring to a crucible;

(2) putting the crucible filled with the mixed powder in the step (1) in N₂Placing the mixture in heating equipment under the atmosphere;

(3) heating the equipment in the step (2) for 10K min^-1The temperature of the crucible is increased from room temperature to 450K at the temperature increasing rate, and the crucible is subjected to heat preservation treatment at the temperature for 20 min;

(4) cooling the equipment in the step (3) for 10K min^-1The crucible is cooled to 273.15K at the cooling rate;

(5) and (5) transferring the sample in the step (4) to an annealing furnace with the temperature of 283.15K, keeping the temperature for 10min, and annealing the glass to obtain the mixed glass.

Fig. 14 shows a pattern printed by the mixed glass prepared in example 6 through a 3D printing device. The mixed powder was placed in a barrel of a 3D printer, and the heating temperature was set at 450K.

FIG. 15 shows the degradation of the mixed glass prepared in example 6 after implantation in an animal mouse model.

Example 7

A method for preparing insulin-loaded amino acid-based glass comprises the following steps:

(1) weighing 50mg of immune active peptide Val-Gln-Pro-Ile-Pro-Tyr powder, placing the powder in a mortar, uniformly grinding the powder, and transferring the powder to a crucible;

(2) putting the crucible filled with the mixed powder in the step (1) in N₂Placing the mixture in heating equipment under the atmosphere;

(3) heating the equipment in the step (2) for 10K min^-1The heating rate of (2) is to raise the crucible from room temperatureHeating to 450K, preserving heat at the temperature for 10min, and then cooling to 330K;

(4) weighing 5mg of insulin powder, placing the insulin powder in a mortar, grinding the insulin powder uniformly, transferring the insulin powder to the crucible in the step (3), stirring the insulin powder uniformly, and preserving heat at the temperature for 10min to melt the insulin powder;

(5) cooling the equipment in the step (4) for 20K min^-1The crucible is cooled to 273.15K at the cooling rate;

(6) and (4) transferring the sample in the step (5) to an annealing furnace with the temperature of 273.15K, keeping the temperature for 20min, and annealing the glass to obtain the insulin-loaded amino acid-based glass.

FIG. 16 is a graph of the biodegradation of insulin-loaded amino acid-based glass prepared in example 7, implanted subcutaneously in mice over time.

FIG. 17 is a graph showing the blood glucose changes in insulin-loaded amino acid-based glass prepared in example 7 after oral gavage in diabetic mice.