CN106267324A - A kind of temperature response type macromolecule bio-medical adhesive and synthetic method thereof - Google Patents

Abstract

The invention provides a temperature-responsive polymer biomedical adhesive and its preparation, belonging to the technical field of adhesive materials. The present invention uses polyhydroxyl or polyamino hyperbranched polymer as initiator, tetrahydrofuran as solvent, amino acid with N-carbonyl internal anhydride at the end as functional monomer, under nitrogen protection, polymerizes at room temperature for 48-72h, after the reaction is over , the crude product was precipitated with a mixed solution of triethylamine-ethanol, and the obtained polymer biomedical adhesive had a bonding strength between 90 and 130 Kpa at 37±0.5°C. In addition, the polymer biomedical adhesive of the present invention uses amino acids required by the human body as raw materials, so it has good biocompatibility, no toxic and side effects, and can be degraded and metabolized in tissues.

Description

A temperature-responsive polymer biomedical adhesive and its synthesis method

technical field

The invention relates to a medical adhesive, in particular to a temperature-responsive polymer biomedical adhesive and a synthesis method thereof, belonging to the fields of polymer materials and biomedical materials.

Background technique

Medical adhesives are a new class of biomedical materials. Various incision adhesives have been used clinically to replace sutures. The advantages are convenience, quickness, no need to remove stitches, and small scars after wound healing. There are many types of medical adhesives, which can be divided into soft tissue adhesives, dental adhesives, bone cement and skin adhesives according to their uses. Adhesives for soft tissue that have been clinically used include α-cyanoacrylate, fibrin, and polyurethane adhesives. However, these adhesives also have some insurmountable shortcomings, such as α-cyanoacrylate, although the bonding time is short and the curing is fast, but the adhesive layer is brittle, low in strength, and formaldehyde will be generated during the decomposition process; although fibrin is biocompatible Sexuality and hemostatic effect are good, but the strength is low and easy to leak. In contrast, polymer hydrogels not only have hemostatic and analgesic effects, but also have good biocompatibility, elasticity and strength. Therefore, the research of polymer hydrogels as soft tissue adhesives has gradually attracted the attention of researchers. Inspired by the mollusk mussel, the catechol functional group is introduced into the polymer chain, and the hydrogel system is formed in situ through chemical cross-linking, which has good tissue adhesion and can be used for soft tissues such as intestines and skin. bonding. Such hydrogel systems that have been reported include modified chitosan/polyethylene glycol, modified chitosan/polyether, chitosan/3,4-dihydroxyphenylpropionic acid, polyethylene glycol/polyethylene glycol, etc. Caprolactone/dihydroxyphenylpropionic acid, citric acid/polyethylene glycol/dopamine, etc. Compared with commercial medical adhesives, the breaking strength and tensile strength have been greatly improved respectively. For example, Mathiowitz et al. reported a polyacrylic acid derivative hydrogel adhesive with dihydroxyphenylalanine side groups. The breaking strength of this new adhesive was tested by the intestinal adhesion test of living rats. and tensile strength increased by 2.5 and 2.8 times, respectively. In addition to dopamine-like structures, guanidine and phosphorylcholine are charged bioorganic functional groups that can interact with histone chains and thus can also be used for medical bonding. Okuro et al. synthesized a water-soluble dendritic molecule with nine guanidine groups, which can gel in situ with tissue protein, which is a typical "molecular glue". Yu et al. modified the end group of dendritic polyethylene glycol with phosphorylcholine fragments, which can form biofilms when bonded, with good biocompatibility and fast wound healing. In addition, Strehin et al. used N-hydroxysuccinimide to esterify the carboxyl group of chondroitin sulfate, then reacted with primary amines, and then used six-armed amino-terminated polyethylene glycol as a cross-linking agent to form a hydrogel, which can be combined with The amino groups of histones are covalently bonded to achieve the purpose of adhesion. Serrero et al. used chitosan and modified starch (oxidized maltodextrin) as adhesives for tissue adhesion. However, these hydrogel adhesives still have many deficiencies such as slow curing, insufficient strength, or easy leakage of body fluids, etc., and further improvement is still needed.

Contents of the invention

The purpose of the present invention is to provide a temperature-responsive polymer biomedical adhesive with fast curing speed, good degradation performance and high bonding strength for the shortcomings of existing medical adhesives;

Another object of the present invention is to provide a method for synthesizing the above-mentioned temperature-responsive polymer biomedical adhesive.

1. Synthesis of temperature-responsive polymer biomedical adhesive

The invention uses polyhydroxyl or polyamino hyperbranched polymer as initiator, tetrahydrofuran as solvent, amino acid with N-carbonyl internal acid anhydride (NCA) at the end as functional monomer, under nitrogen protection, polymerizes at room temperature for 48~72h , After the reaction, the crude product was obtained by precipitation with a mixed solution of triethylamine-ethanol.

The amino acid with N-carbonyl internal anhydride (NCA) at the end is L-glutamic acid-β-diethylene glycol monomethyl ether ester-N-carbonyl internal anhydride (Glu-EG ₂ -NCA), L-3, 4-Dihydroxybenzylaminoacetic acid-N-carbonyl anhydride (DOPA-NCA), arginine-N-carbonyl anhydride (Arg-NCA), cysteine-N-carbonyl anhydride (Cys-NCA ) or at least two of ε-N-acrylamide lysine-N-carbonyl anhydride (AM-Lys-NCA).

The polyhydroxyl hyperbranched polymer is a hyperbranched polyhydroxyl polymer obtained by reacting 1,4-butanediol and metal potassium to initiate glycidol ring opening to obtain hyperbranched polyglycidyl ether, and then adding metal potassium to react Potassium alcohol.

For the synthesis of hyperbranched polyglycidyl ether, see literature S. Roller, H. Zhou, R. Haag, Mol Divers2005, 9, 305.), its structure is:

The structure of hyperbranched polyol potassium alkoxide is:

.

Described polyamino hyperbranched polymer is hyperbranched polyamide-amine, and its product trade mark: CYD-130A.

The molar weight of the initiator and the functional monomer (total amount) is 1:620-1:1030.

The functional monomers are L-glutamic acid-β-diethylene glycol monomethyl ether ester-N-carbonyl internal anhydride (Glu-EG ₂ -NCA), L-3,4-dihydroxybenzylaminoacetic acid -N-carbonyl internal anhydride (DOPA-NCA), the molar ratio of the two is 1:0.9~1:1.1.

The functional monomers are L-glutamic acid-β-diethylene glycol monomethyl ether ester-N-carbonyl internal anhydride (Glu-EG ₂ -NCA), L-3,4-dihydroxybenzylaminoacetic acid -N-carbonyl internal anhydride (DOPA-NCA) and arginine-N-carbonyl internal anhydride (Arg-NCA), the molar ratio of the three is 1:0.9:0.58~1:1.1:0.63.

The functional monomers are L-glutamic acid-β-diethylene glycol monomethyl ether ester-N-carbonyl internal anhydride (Glu-EG ₂ -NCA), L-3,4-dihydroxybenzylaminoacetic acid -N-carbonyl anhydride (DOPA-NCA), arginine-N-carbonyl anhydride (Arg-NCA), cysteine-N-carbonyl anhydride (Cys-NCA), the molar ratio of the four is 1:0.9:0.58:0.58~1:1.1:0.63:0.63.

The functional monomers are L-glutamic acid-β-diethylene glycol monomethyl ether ester-N-carbonyl internal anhydride (Glu-EG ₂ -NCA), L-3,4-dihydroxybenzylaminoacetic acid -N-carbonyl anhydride (DOPA-NCA), arginine-N-carbonyl anhydride (Arg-NCA), ε-N-acrylamide lysine-N-carbonyl anhydride (AM-Lys-NCA) , the molar ratio of the four is 1:0.9:0.58:0.58~1:1.1:0.63:0.64.

2. Performance of temperature-responsive biomedical adhesives

1. Temperature sensitivity test

Test method: The lowest critical solution temperature (LCST) is an important characterization parameter for the temperature-sensitive properties of polymers. By measuring the curve of the light transmittance of the polymer solution as a function of temperature, the temperature corresponding to when the light transmittance drops to 50% of the initial light transmittance is defined as its LCST value. The temperature-sensitive properties of polymers are based on the relative balance between the hydrophilicity and hydrophobicity of polymers. Prepare a polymer aqueous solution with a concentration of 0.3 mg/mL, and measure the light transmittance curve of the solution at different temperatures by an ultraviolet-visible spectrophotometer (UV-VIS) at 500 nm to obtain its LCST value.

Measurement results: By measuring the curve of the light transmittance of the polymer solution as a function of temperature, its lowest critical solution temperature (LCST) can be obtained. Figure 1 is an adhesive solution prepared with hyperbranched polyol potassium alkoxide as an initiator The light transmittance versus temperature curve. From Figure 1 we can see that the LCST value of this kind of polymer is about 33 ℃. Fig. 2 is the light transmittance versus temperature curve of the adhesive solution prepared with hyperbranched polyamidoamine as the initiator. From Fig. 2 we can see that the LCST value of this kind of polymer is about 31 ℃.

2. Bonding performance test

Bonding strength test method: pigskin is first washed with 0.9% sodium chloride solution, and then soaked overnight in PBS buffer solution with pH=7.4 for testing. The resulting polymer was dissolved in PBS buffer at pH = 7.4 to prepare a mass fraction of 10%, and then it was dissolved in 0.06 mg/mL horseradish peroxidase and 0.06% peroxidase In the hydrogen solution, use it on the surface of two pieces of pigskin respectively, then press the two pieces of pigskin together, and give a certain pressure at the same time, adopt the ASTM test standard, and carry out the pigskin sticking together at a rate of 10mm/min until the two pieces of pigskin are pulled Until it is opened, record the data and measure three groups in parallel.

Test results: Figure 3 shows the test results of the bonding strength of the adhesive solution prepared in Examples 1-4. From the test results in Figure 3 and Figure 4, it was found that the bond strength of the prepared adhesive gradually increased with the increase of the test time. At the same time, with the increase of the types of functional monomers introduced, the bonding strength of the obtained adhesive also gradually increased. This is because the guanidinium cation in arginine can overlap with the carboxyl group in the protein to generate crosslinking, and the sulfhydryl group in the cysteine can undergo oxidation reaction with the sulfhydryl group in the protein to form a disulfide bond. The sulfhydryl group in amino acid and the double bond in acrylamide lysine can undergo a click reaction to increase the bonding strength.

Fig. 5 shows the test results of the bonding strength of the adhesive solutions prepared in Examples 1-4 under the conditions of 25°C and 37°C respectively. Fig. 6 shows the test results of the bonding strength of the adhesive solutions prepared in Examples 5-8 under the conditions of 25°C and 37°C respectively. From the test results in Figure 5 and Figure 6, it is found that the curing rate of the adhesive at 37°C is significantly faster than that at 25°C. It is due to the introduction of diethylene glycol monomethyl ether into the substance that the adhesive we obtained has temperature responsiveness, so that when the temperature reaches 37°C, the adhesive solidifies quickly and can prevent the leakage of body fluids. And 37 ℃ is normal human body temperature, so this invention is suitable as medical adhesive.

In summary, the present invention uses polyhydroxy hyperbranched potassium alkoxide or hyperbranched polyamide-amine as initiator, and L-glutamic acid-β-diethylene glycol monomethyl ether ester (Glu-EG ₂ ) and 3 , 4-dihydroxyphenylalanine (DOPA) residues as the adhesive group hyperbranched polymer adhesive HPGMD, on the basis of the adhesive HPGMD, in order to improve the curing rate and bond strength, the introduction of arginine segment to obtain the adhesive HPGMDA/PAMAMGMD; in order to further improve the bonding strength, on the basis of the adhesive HPGMDA/PAMAMGMDA, a cysteine segment and an acrylylated lysine segment were respectively introduced to obtain the adhesive HPGMDAC /PAMAMGMDAC and HPGMDAL/PAMAMGMDAL, this adhesive can be mixed immediately at the time of use. The performance of the above-mentioned adhesives is superior to most similar biomimetic polymer adhesives reported in the literature. This is because the guanidine group in arginine can cross-link with the carboxyl group in the protein, and the sulfhydryl group in the cysteine can undergo oxidation reaction with the sulfhydryl group in the protein to form a disulfide bond. A click reaction can occur with the double bond in acrylamide lysine to increase its tensile strength. In addition, this adhesive is made of amino acids required by the human body, so it has good biocompatibility, no toxic side effects, and can be degraded and metabolized in tissues.

Description of drawings

Figure 1 is a graph showing the light transmittance versus temperature of an adhesive solution prepared with hyperbranched polyol potassium alkoxide as an initiator. Fig. 2 is the light transmittance versus temperature curve of the adhesive solution prepared with hyperbranched polyamidoamine as the initiator.

Fig. 3 is the bonding strength test result of the adhesive solution prepared in Examples 1-4.

Fig. 4 is the bonding strength test result of the adhesive solution prepared in Example 5-8.

Fig. 5 shows the test results of the bonding strength of the adhesive solutions prepared in Examples 1-4 under the conditions of 25°C and 37°C respectively.

Fig. 6 shows the test results of the bonding strength of the adhesive solutions prepared in Examples 5-8 under the conditions of 25°C and 37°C respectively.

detailed description

The synthesis method and performance of the temperature-responsive biomedical adhesive of the present invention will be further described below through specific examples.

Example 1: Preparation of HP(DOPA-Glu-EG ₂ )

Preparation of Glu-EG ₂ : Add 2.5 g (16.99 mmol) of glutamic acid and 15 mL (129.21 mmol) of di Ethylene glycol monomethyl ether, 2 mL of concentrated sulfuric acid was added to a constant pressure dropping funnel, and slowly added to a three-necked flask under nitrogen protection at 0 °C and reacted for 12 h under an ice bath to obtain a crude product. Pour the crude product into a mixture of 50 mL of triethylamine and 50 mL of ethanol to obtain a white precipitate, centrifuge, dissolve the white precipitate in 10 mL of methanol, then pour the solution into the white precipitate in 100 mL of ether, and centrifuge to obtain The white precipitated product was vacuum-dried to obtain a white solid, L-glutamic acid-β-diethylene glycol monomethyl ether ester (Glu-EG ₂ ), with a yield of 55%.

Preparation of Glu-EG2-NCA: Add 1 g (4.02 mmol) Glu-EG ₂ and anhydrous-treated Add 30 mL of tetrahydrofuran to the constant pressure dropping funnel and add 10 mL of tetrahydrofuran solution dissolved with 1.94 g (6.54 mmol) of triphosgene. Raise the temperature to about 50 °C, and slowly add the triphosgene solution in the constant pressure dropping funnel into the three-necked flask under magnetic stirring. React until the solution becomes a light yellow transparent solution and then react for 30 min. At this time, nitrogen gas is passed to bring the residual triphosgene and HCl gas into the absorption device containing saturated NaOH solution. After tail gas absorption treatment, the reaction liquid was concentrated, an appropriate amount of n-hexane was added, and left to stand, a light yellow solid was precipitated, and the crude product was obtained by filtration. The crude product was recrystallized from n-hexane and ethyl acetate to obtain white crystals which were L-glutamic acid-β-diethylene glycol monomethyl ether ester-N-carbonyl internal acid anhydride (Glu-EG ₂ -NCA).

Preparation of DOPA-NCA: Add 0.5 g (2.54 mmol) of 3,4-dihydroxyphenylalanine (DOPA) to a 50 mL three-necked flask equipped with a magnetic stirrer, thermometer, constant pressure dropping funnel, and nitrogen protection device And 20 mL of tetrahydrofuran that has been anhydrous treated, add 10 mL of tetrahydrofuran solution dissolved with 0.5 g (1.69 mmol) of triphosgene into the constant pressure dropping funnel. Raise the temperature to about 50°C, and slowly add the triphosgene solution in the constant pressure dropping funnel into the three-necked flask under magnetic stirring. React until the solution becomes a light yellow transparent solution and then react for 30 min. At this time, nitrogen gas is passed to bring residual phosgene and HCl gas into the absorption device containing saturated NaOH solution. After tail gas absorption treatment, the reaction solution was concentrated, and an appropriate amount of n-hexane was added. After standing still, a light yellow oily substance precipitated out. After repeated precipitation for several times, the light yellowish oily substance obtained was L-3,4-dihydroxybenzylaminoacetic acid-N-carbonyl internal acid anhydride (DOPA-NCA).

Preparation of HP(DOPA-Glu-EG ₂ ): Add 1.65 g (6.00 mmol) L-glutamic acid- β-diethylene glycol monomethyl ether ester-N-carbonyl internal acid anhydride (Glu-EG ₂ -NCA) and 10 mL of anhydrous-treated tetrahydrofuran, add 3 mL of anhydrous THF-dissolved initiator into the constant-pressure dropping funnel Slowly add 0.05 g (0.0192 mmol) of hyperbranched polyglycidyl ether into a three-necked flask, and react for 72 hours to obtain a light yellow viscous substance. Then add 1.34 g (6.00 mmol) of L-3,4-dihydroxybenzylaminoacetic acid-N-carbonyl internal anhydride (DOPA-NCA) dissolved in 10 mL of anhydrous tetrahydrofuran into the constant pressure dropping funnel, and slowly add to In a three-necked flask, react for 72 hours to obtain a light yellow viscous substance which is the crude product. Absolute ethanol was added to the crude product to produce a light yellow precipitate, and the obtained light yellow oil was HP(DOPA-Glu-EG ₂ ).

Through the detection of the temperature sensitivity and bonding performance of the polymer, it was found that the response temperature was 37±0.5°C, and the bonding strength was about 90 Kpa.

Example 2: Preparation of HP (Arg-DOPA-Glu-EG ₂ )

Preparation of Glu-EG ₂ : Same as Example 1;

Preparation of Glu-EG ₂ -NCA: Same as Example 1;

Preparation of DOPA-NCA: same as Example 1;

Preparation of HP (DOPA-Glu-EG ₂ ): Same as Example 1;

Preparation of Arg-NCA: Add 6.6 g (21.43 mmol) of benzyloxycarbonyl arginine and 30 mL of solvent tetrahydrofuran into a 100 mL three-necked flask equipped with a magnetic stirrer, thermometer, constant pressure dropping funnel and nitrogen protection device. Add 5 mL of tetrahydrofuran solution containing 1 mL of phosphorus tribromide (10.64 mmol) into the constant pressure dropping funnel. Under nitrogen protection, it was added dropwise into a three-necked flask under ice-cooling, and then reacted at room temperature for 16 hours. After the reaction, the reaction solution was separated into layers, and the tetrahydrofuran solution in the upper layer was poured out, and the oil in the lower layer was washed repeatedly with a large amount of tetrahydrofuran to obtain a colorless and transparent oil that was arginine-N-carbonyl anhydride (Arg-NCA).

Preparation of HP(Arg-DOPA-Glu-EG ₂ ): Add 0.76 g (3.80 mmol) of arginine-N-carbonyl anhydride (Arg-NCA) dissolved in 10 mL of anhydrous tetrahydrofuran into a constant pressure dropping funnel, Slowly add to the steps In a three-necked flask, the reaction was carried out for three days to obtain a light yellow viscous substance which was the crude product. The crude product was washed with absolute ethanol, and the obtained pale yellow oil was HP(Arg-DOPA-Glu-EG ₂ ).

Through the detection of the temperature sensitivity and bonding performance of the polymer, it was found that the response temperature was 37±0.5°C, and the bonding strength was about 100 Kpa.

Example 3: Preparation of HP (Cys-Arg-DOPA-Glu-EG ₂ )

Preparation of Glu-EG ₂ : Same as Example 1;

Preparation of Glu-EG ₂ -NCA: Same as Example 1;

Preparation of DOPA-NCA: same as Example 1;

Preparation of HP (DOPA-Glu-EG ₂ ): Same as Example 1;

Preparation of Arg-NCA: same as Example 2;

Preparation of HP (Arg-DOPA-Glu-EG ₂ ): same as Example 2;

Preparation of Cys-NCA: Add 1 g (8.25 mmol) cysteine and 30 mL of anhydrous-treated tetrahydrofuran into a 100 mL three-necked flask equipped with a magnetic stirrer, a thermometer, a constant pressure dropping funnel and a nitrogen protection device, Add 10 mL of tetrahydrofuran solution with 1.47 g (4.96 mmol) of triphosgene dissolved in the constant pressure dropping funnel. Raise the temperature to about 50 °C, and slowly add the triphosgene solution in the constant pressure dropping funnel into the three-necked flask under magnetic stirring. React until the solution becomes a light yellow transparent solution and then react for 30 min. At this time, nitrogen gas is passed to bring residual phosgene and HCl gas into the absorption device containing saturated NaOH solution. After tail gas absorption treatment, the reaction solution was concentrated, and an appropriate amount of n-hexane was added. After standing still, a light yellow solid precipitated out, and the crude product was obtained by filtration. The crude product was recrystallized from n-hexane and ethyl acetate to obtain white crystals, cysteine-N-carbonyl anhydride (Cys-NCA).

Preparation of HP (Cys-Arg-DOPA-Glu-EG ₂ ): Add 0.56 g (3.80 mol), slowly added to the step In a three-necked flask, the reaction was carried out for three days to obtain a light yellow viscous substance which was the crude product. The crude product was washed with absolute ethanol to obtain a pale yellow oil which was HP (Cys-Arg-DOPA-Glu-EG ₂ ).

Through the detection of the temperature sensitivity and bonding performance of the polymer, it is found that the response temperature is 37±0.5°C, and the bonding strength is about 108Kpa.

Example 4: Preparation of HP (AM-Lys-Arg-DOPA-Glu-EG ₂ ):

Preparation of Glu-EG ₂ : Same as Example 1;

Preparation of Glu-EG ₂ -NCA: Same as Example 1;

Preparation of DOPA-NCA: same as Example 1;

Preparation of HP (DOPA-Glu-EG ₂ ): Same as Example 1;

Preparation of Arg-NCA: same as Example 2;

Preparation of HP (Arg-DOPA-Glu-EG ₂ ): same as Example 2;

Preparation of AM-Lys-NCA:

Acrylamide lysine copper salt: Add 1 g (5.47 mmol) lysine hydrochloride to a 50 mL round-bottomed flask equipped with a magnetic stirrer and a thermometer, and add 12 mL of water at the same time, reflux at 90 ° C, and dissolve 0.66 g (2.99 mmol) basic copper carbonate was slowly added to the solution of lysine hydrochloride, stirred for 10 min, then cooled, and the insoluble residue was filtered, 5.8 mL of acetone was added to the filtrate, and then 2.74 mL of 2 mol /L of potassium hydroxide solution, the mixed solution was added to a 100mL three-necked flask equipped with a magnetic stirrer, a thermometer, a constant pressure dropping funnel and a nitrogen protection device, and 0.552 mL (6.79mmol) of acryloyl chloride and 3.08 mL of 2 mol/L potassium hydroxide solution were slowly dropped into the reaction system under an ice bath, reacted for 12 h, and filtered the precipitate of acrylamide lysine copper complex, Wash with water, methanol and ether successively. The resulting blue solid is acrylamide lysine copper complex.

Preparation of acrylamide lysine: 0.21 g (0.43 mmol) of the copper acrylamide lysine complex obtained in the previous step was added to a 100 mL round-bottomed flask equipped with a magnetic stirrer, a thermometer, and a nitrogen protection device. Add 3 mL of water and 3 mL of chloroform solution dissolved with 0.079 g (0.54 mmol) of 8-hydroxyquinoline, stir at room temperature for about 12 h, a green precipitate will appear in the chloroform layer, remove the precipitate by filtration, and transfer the filtrate to the In a liquid funnel, remove the chloroform layer, then wash with chloroform for 3-4 times, concentrate the remaining water layer to a white solid, then recrystallize with tetrahydrofuran and water to obtain a white crystal that is acrylamide lysine, The yield was 45%.

Preparation of AM-Lys-NCA: Add 2 g (10.00 mmol) of acrylamide lysine obtained in the previous step to a 100 mL three-necked flask equipped with a magnetic stirrer, a thermometer, a constant pressure dropping funnel, and a nitrogen protection device. After anhydrous treatment of 40 mL of tetrahydrofuran, 0.011 g (0.1 mmol) of hydroquinone, a polymerization inhibitor, was added at the same time, and 20 mL of tetrahydrofuran solution in which 1.78 g (5.99 mmol) of triphosgene was dissolved was added to a constant pressure dropping funnel. Raise the temperature to about 50 °C, and slowly add the triphosgene solution in the constant pressure dropping funnel into the three-necked flask under magnetic stirring. React until the solution becomes a light yellow transparent solution and then react for 30 min. At this time, nitrogen gas is passed to bring residual phosgene and HCl gas into the absorption device containing saturated NaOH solution. After tail gas absorption treatment, the reaction solution was concentrated, and an appropriate amount of n-hexane was added. After standing still, a light yellow solid precipitated out, and the crude product was obtained by filtration. The crude product was recrystallized from n-hexane and ethyl acetate to obtain white crystals, ε-N-acrylamide lysine-N-carbonyl anhydride (AM-Lys-NCA), with a yield of 60%.

Preparation of HP (AM-Lys-Arg-DOPA-Glu-EG ₂ ): Add ε-N-acrylamide lysine-N-carbonyl internal anhydride (AM -Lys- NCA) 0.86 g (3.84mmol), slowly added to step In a three-necked flask, react for three days to obtain a light yellow viscous substance which is the crude product. The crude product is washed with absolute ethanol to obtain a light yellow oil which is HP(AM-Lys-Arg-DOPA-Glu-EG ₂ ). Its structural formula is as follows:

Through the detection of the temperature sensitivity and bonding performance of the polymer, it is found that the response temperature is 37±0.5°C, and the bonding strength is about 117Kpa.

Example 5: Preparation of PAMAM (DOPA-Glu-EG ₂ )

Preparation of Glu-EG ₂ : Same as Example 1;

Preparation of Glu-EG2-NCA: same as Example 1;

Preparation of DOPA-NCA: same as Example 1;

Preparation of PAMAM (DOPA-Glu-EG ₂ ): Add 0.62 g (2.25 mmol) L-glutamic acid- β-Diethylene glycol monomethyl ether ester-N-carbonyl internal acid anhydride (Glu-EG2-NCA) and 10 mL of tetrahydrofuran through anhydrous treatment were added to the constant pressure dropping funnel with 3 mL of anhydrous tetrahydrofuran dissolved in the initiator overrun Add 0.05 g (0.0072 mmol) of polyamidoamine to a three-necked flask slowly, and react for three days to obtain a light yellow viscous substance. Then add 0.51 g (2.25 mmol) of L-3,4-dihydroxybenzylaminoacetic acid-N-carbonyl internal anhydride (DOPA-NCA) dissolved in 10 mL of anhydrous tetrahydrofuran into the constant pressure dropping funnel, and slowly add to In a three-necked flask, react for three days to obtain a light yellow viscous substance which is the crude product. Absolute ethanol was added to the crude product to produce a light yellow precipitate, and the obtained light yellow oil was PAMAM (DOPA-Glu-EG ₂ ). By testing the temperature sensitivity and bonding performance of the polymer, it was found that the response temperature was 37±0.5°C, and the bonding strength was about 93 Kpa.

Example 6: Preparation of PAMAM (Arg-DOPA-Glu-EG ₂ )

Preparation of Glu-EG ₂ : Same as Example 1;

Preparation of Glu-EG2-NCA: same as Example 1;

Preparation of DOPA-NCA: same as Example 1;

Preparation of PAMAM (DOPA-Glu-EG ₂ ): Same as Example 5;

Preparation of Arg-NCA: same as Example 2;

Preparation of PAMAM (Arg-DOPA-Glu-EG ₂ ): Add 0.29 g (1.44 mmol) of arginine-N-carbonyl anhydride (Arg-NCA) dissolved in 10 mL of anhydrous tetrahydrofuran into a constant pressure dropping funnel, Slowly add to the steps In a three-necked flask, the reaction was carried out for three days to obtain a light yellow viscous substance which was the crude product. The crude product was washed with absolute ethanol, and the obtained pale yellow oil was PAMAM (Arg-DOPA-Glu-EG ₂ ).

Through the detection of the temperature sensitivity and bonding performance of the polymer, it was found that the response temperature was about 37±0.5°C, and the bonding strength was about 102 Kpa.

Example 7: Preparation of PAMAM (Cys-Arg-DOPA-Glu-EG ₂ )

Preparation of Glu-EG ₂ : Same as Example 1;

Preparation of Glu-EG ₂ -NCA: Same as Example 1;

Preparation of DOPA-NCA: same as Example 1;

Preparation of PAMAM (DOPA-Glu-EG ₂ ): Same as Example 5;

Preparation of Arg-NCA: same as Example 2;

Preparation of PAMAM (Arg-DOPA-Glu-EG ₂ ): Same as Example 6;

Preparation of Cys-NCA: Add 1 g (8.25 mmol) cysteine and 30 mL of anhydrous-treated tetrahydrofuran into a 100 mL three-necked flask equipped with a magnetic stirrer, a thermometer, a constant pressure dropping funnel and a nitrogen protection device, Add 10 mL of tetrahydrofuran solution with 1.47 g (4.96 mmol) of triphosgene dissolved in the constant pressure dropping funnel. Raise the temperature to about 50 °C, and slowly add the triphosgene solution in the constant pressure dropping funnel into the three-necked flask under magnetic stirring. React until the solution becomes a light yellow transparent solution and then react for 30 min. At this time, nitrogen gas is passed to bring residual phosgene and HCl gas into the absorption device containing saturated NaOH solution. After tail gas absorption treatment, the reaction solution was concentrated, and an appropriate amount of n-hexane was added. After standing still, a light yellow solid precipitated out, and the crude product was obtained by filtration. The crude product was recrystallized from n-hexane and ethyl acetate to obtain white crystals, cysteine-N-carbonyl anhydride (Cys-NCA).

Preparation of PAMAM (Cys-Arg-DOPA-Glu-EG ₂ ): Add 0.22 g (Cys-NCA) dissolved in 10 mL of anhydrous tetrahydrofuran into a constant pressure dropping funnel (Cys-NCA) mmol), slowly added to the step In a three-necked flask, the reaction was carried out for three days to obtain a light yellow viscous substance which was the crude product. The crude product was washed with absolute ethanol to obtain a light yellow oily substance which was PAMAM (Cys-Arg-DOPA-Glu-EG ₂ ).

Through the detection of the temperature sensitivity and bonding performance of the polymer, it is found that the response temperature is 37±0.5°C, and the bonding strength is about 110Kpa.

Example 8: Preparation of PAMAM (AM-Lys-Arg-DOPA-Glu-EG ₂ ):

Preparation of Glu-EG ₂ : Same as Example 1;

Preparation of Glu-EG ₂ -NCA: Same as Example 1;

Preparation of DOPA-NCA: same as Example 1;

Preparation of PAMAM (DOPA-Glu-EG ₂ ): Same as Example 5;

Preparation of Arg-NCA: same as Example 2;

Preparation of PAMAM (Arg-DOPA-Glu-EG ₂ ): Same as Example 6;

Preparation of AM-Lys-NCA: same as Example 4;

Preparation of PAMAM (AM-Lys-Arg-DOPA-Glu-EG ₂ ): Add ε-N-acrylamide lysine-N-carbonyl internal acid anhydride (AM -Lys- NCA) 0.33 g (1.44mmol), slowly added to step In a three-necked flask, react for three days to obtain a light yellow viscous substance which is the crude product. The crude product is washed with absolute ethanol to obtain a light yellow oil which is PAMAM (AM-Lys-Arg-DOPA-Glu-EG ₂ ).

Through the detection of the temperature sensitivity and bonding performance of the polymer, it is found that its response temperature is 37±0.5°C, and the bonding strength is about 120Kpa.

Claims (10)

1. a preparation method for temperature response type macromolecule bio-medical adhesive, is with polyhydroxy or polyamino hyperbranched poly Compound is initiator, with oxolane as solvent, has the aminoacid of N-carbonyl inner-acid anhydride as function monomer with end, and nitrogen is protected Protecting, react 48 ~ 72h in polymerized at room temperature, after reaction terminates, the mixed solution of crude product triethylamine-ethanol precipitates and obtains.

2. the preparation method of temperature response type macromolecule bio-medical adhesive as claimed in claim 1, it is characterised in that: described End has the aminoacid of N-carbonyl inner-acid anhydride to be Pidolidone-β-diethylene glycol monomethyl ether ester-N-carbonyl inner-acid anhydride, L-3,4-bis- Hydroxyphenylmethyl glycine-N-carbonyl inner-acid anhydride, arginine-N-carbonyl inner-acid anhydride, cysteine-N-carbonyl inner-acid anhydride or ε- At least two in N acrylamide lysine-N-carbonyl inner-acid anhydride.

3. the preparation method of temperature response type macromolecule bio-medical adhesive as claimed in claim 1, it is characterised in that: described Polyhydroxy dissaving polymer is to react with BDO and metallic potassium to cause epoxy prapanol open loop, obtains hyperbranched poly and shrinks Glycerin ether, adds metallic potassium, the hyperbranched polyhydroxylated polymer potassium alcoholate that reaction obtains.

The preparation method of a kind of temperature response type macromolecule bio-medical adhesive the most as claimed in claim 1, it is characterised in that: Described polyamino dissaving polymer is ultrabranching polyamide-amine.

5. the preparation method of any temperature response type macromolecule bio-medical adhesive as described in claim 1-4, it is special Levy and be: described initiator is 1:620 ~ 1:1030 with the mole of function monomer.

6. the preparation method of any temperature response type macromolecule bio-medical adhesive as described in claim 1-4, it is special Levy and be: described function monomer is Pidolidone-β-diethylene glycol monomethyl ether ester-N-carbonyl inner-acid anhydride, L-3,4-dihydroxy benzenes During methyl aminoacetic acid-N-carbonyl inner-acid anhydride, the mol ratio of the two is 1:0.9 ~ 1:1.1.

7. the preparation method of any temperature response type macromolecule bio-medical adhesive as described in claim 1-4, it is special Levy and be: described function monomer is Pidolidone-β-diethylene glycol monomethyl ether ester-N-carbonyl inner-acid anhydride, L-3,4-dihydroxy benzenes When methyl aminoacetic acid-N-carbonyl inner-acid anhydride, arginine-N-carbonyl inner-acid anhydride, the mol ratio of three is 1:0.9:0.58 ~ 1: 1.1:0.63。

8. the preparation method of any temperature response type macromolecule bio-medical adhesive as described in claim 1-4, it is special Levy and be: function monomer is Pidolidone-β-diethylene glycol monomethyl ether ester-N-carbonyl inner-acid anhydride, L-3,4-dihydroxy benzenes methyl When glycine-N-carbonyl inner-acid anhydride, arginine-N-carbonyl inner-acid anhydride, cysteine-N-carbonyl inner-acid anhydride, four mole Ratio is 1:0.9:0.58:0.58 ~ 1:1.1:0.63:0.63.

9. the preparation method of any one temperature response type macromolecule bio-medical adhesive as described in claim 1-4, it is special Levy and be: function monomer is Pidolidone-β-diethylene glycol monomethyl ether ester-N-carbonyl inner-acid anhydride, L-3,4-dihydroxy benzenes methyl When glycine-N-carbonyl inner-acid anhydride, arginine-N-carbonyl inner-acid anhydride, ε-N acrylamide lysine-N-carbonyl inner-acid anhydride, The mol ratio of four is 1:0.9:0.58:0.58 ~ 1:1.1:0.63:0.64.

10. the temperature response type macromolecule bio-medical adhesive that as claimed in claim 1 prepared by method, it is characterised in that: temperature Degree response is when 37 ± 0.5 DEG C, and bond strength is between 90 ~ 130 Kpa.