KR20200129589A - Dopamine functionalized polyurethane based polymer for tissue adhesive, method of manufacturing the polymer, and tissue adhesive having the polymer - Google Patents

Abstract

Disclosed is a polyurethane-based polymer compound for a biological tissue adhesive, comprising a first monomer represented by chemical formula 1 and a second monomer represented by chemical formula 2. Dopamine and the polyurethane-based polymer compound for a biological tissue adhesive containing the same are not cytotoxic, and thus can be applied to a human body.

Description

Polyurethane-based polymer compound for biotissue adhesive containing functional dopamine, a method for manufacturing the same, and biotissue adhesive containing the same BACKGROUND OF THE INVENTION

The present invention relates to a polyurethane-based polymer compound applicable to suturing or covering damaged tissue areas or surgical sites, a method of manufacturing the same, and a biological tissue adhesive comprising the same.

To suture or cover damaged tissue areas or surgical areas, conventional fibrous sutures, metal staples, and the like have been generally used.

However, these have decisive problems such as damage to surrounding tissues, secondary infection, and scar formation, and furthermore, metal staples are inherently disadvantageous with a psychological burden of removing metal staples after treatment.

Biotissue adhesive using various polymer materials such as poly (cyanoacrylate), polyethylene glycol (PEG), and polyurethane (polyurethane, PU) to replace recent fiber sutures and metal staples. Much research is being conducted on the material.

In particular, polyurethane polymer materials are excellent in biocompatibility and are physically and chemically stable, and thus are attracting much attention as candidate materials for biotissue adhesives. Polyurethane containing an aromatic isocyanate-terminated prepolymer has excellent reactivity and short curing time, which satisfies some of the characteristics of biological tissue adhesives, but toxic substances are removed through hydrolysis of urethane due to the formation of diamine. There is a problem to generate.

One object of the present invention is to provide a polyurethane-based polymer compound for a biological tissue adhesive having excellent adhesion to biological tissues, mechanical strength, and non-cytotoxic properties by grafting dopamine onto a backbone.

Another object of the present invention is to provide a method for synthesizing the polyurethane-based polymer compound for the biological tissue adhesive.

Another object of the present invention is to provide a biotissue adhesive comprising the polyurethane-based polymer compound.

The polyurethane-based polymer compound for a biotissue adhesive according to an embodiment of the present invention includes a first monomer of Formula 1 and a second monomer of Formula 2 below.

[Formula 1]

[Formula 2]

In Formulas 1 and 2, R 1 is an alkene functional group having 1 to 25 carbon atoms, R 2 is an aliphatic functional group having 5 or more and 15 or less carbon atoms, R 3 is an aliphatic functional group having 5 or more and 15 or less carbon atoms, and a and b are each It is an integer of 1 or more and 3 or less independently.

In one embodiment, R2 may include any one selected from the group consisting of the following Formulas 3-1 to 3-3.

[Formula 3-1]

[Chemical Formula 3-2]

[Chemical Formula 3-3]

In one embodiment, R3 may include a functional group represented by Formula 4.

[Formula 4]

In one embodiment, the polyurethane-based polymer compound may have a molecular structure represented by Formula 5 below.

[Formula 5]

In Formula 5, m and n are each independently an integer of 1 or more and 3 or less.

In one embodiment, the ratio of m and n may be 1:1 to 1:3.

In one embodiment, the polyurethane-based polymer compound may have a molecular weight of about 1500 or more and 3000 g/mol or less.

The method for synthesizing a polyurethane-based polymer compound for a biotissue adhesive according to an embodiment of the present invention can synthesize a polyurethane-based polymer compound having a molecular structure of the following formula (5), and a dopamine-functionalized azide compound ( a first step of synthesizing dopamine functionalized azide); And a second step of mixing and reacting an ethylene glycol compound of Formula 7 below, a diisocyanate compound of Formula 8, a propargyl diol compound of Formula 9, and the dopamine-functionalized azide compound. Include.

[Formula 5]

[Formula 6]

[Formula 7]

[Formula 8]

[Formula 9]

In Formulas 5 to 8, R1 is an alkene functional group having 1 to 25 carbon atoms, R2 is an aliphatic functional group having 5 or more and 15 or less carbon atoms, R3 is an aliphatic functional group having 5 or more and 15 or less carbon atoms, and a and b are each Independently, it is an integer of 1 or more and 3 or less, and m and n are each independently an integer of 1 or more and 3 or less.

In one embodiment, the dopamine-functionalized azide compound of Formula 6 may include the compound of Formula 6-1.

[Formula 6-1]

In one embodiment, the first step is synthesized azido propanol by reacting 3-bromo-1-propanol and sodium azide according to Scheme 1 below. Step to do; Synthesizing a first intermediate compound (1) by reacting the azide propanol, glutaric anhydride, and dimethylaminopyridine (DMAP) according to Scheme 2 below; Synthesizing a second intermediate compound (2) by reacting the first intermediate compound, NHS (N-hydroxysuccinimide) and DCC (N,N'-Dicyclohexylcarbodiimide) according to Scheme 3 below; And synthesizing the compound of Formula 6-1 by reacting the second intermediate compound, Dopamine hydrochloride, and Triethylamine according to Scheme 4 below.

[Scheme 1]

[Scheme 2]

[Scheme 3]

[Scheme 4]

In one embodiment, the diisocyanate compound of Formula 8 may include any one selected from the group consisting of compounds of Formulas 8-1 to 8-3 below.

[Chemical Formula 8-1]

[Formula 8-2]

[Chemical Formula 8-3]

In one embodiment, the propargyl diol compound of Formula 9 is sodium hydride, diethyl-2-methylmalonate, and propargyl bromide according to Scheme 5 below. ) To synthesize a third intermediate compound (3); And reacting the third intermediate compound and lithium aluminum hydride according to Scheme 6 below to synthesize the propargyl diol compound of Formula 9.

[Scheme 5]

[Scheme 6]

In one embodiment, in the second step, based on 1 mol of the ethylene glycol compound, the diisocyanate compound (diisocyanate) is mixed in a ratio of 5 mol or more and 8 mol or less, and the propargyl diol compound ( propargyl diol) is mixed in a ratio of 3 mol or more and 5 mol or less, and the dopamine-functionalized azide compound may be mixed in a ratio of 1 mol or more and 2 mol or less.

According to the polyurethane-based polymer compound for a biological tissue adhesive of the present invention, the catechol group of dopamine grafted on the polyurethane backbone has excellent reactivity to biological tissues containing moisture, protein, etc., so that the biological tissue of the present invention Polyurethane-based polymer compounds for adhesives may have excellent adhesion properties to living tissues. In addition, the dopamine and the polyurethane-based polymer compound for a biological tissue adhesive containing the same is not cytotoxic and thus can be applied to the human body.

1 and 2 are 1H NMR analysis and FT-IR analysis results for azido propanol, intermediate compound (1), intermediate compound (2), and DFA.
3 is a 1H NMR analysis result for propargyl diol.
4 and 5 are graphs showing cell viability of DFA and polyurethane-based polymer compounds of Examples 1 to 3.
6A and 6B are H&E staining images of an experimental rat model for the polymer compound samples of Examples 1 to 3.
7A and 7B are immunostaining images of an experimental rat model for the polymer compound samples of Examples 1 to 3.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. In the present invention, various modifications may be made and various forms may be applied, and specific embodiments will be illustrated in the drawings and described in detail in the text. However, this is not intended to limit the present invention to a specific form of disclosure, it should be understood to include all changes, equivalents, and substitutes included in the spirit and scope of the present invention. In describing each drawing, similar reference numerals have been used for similar elements.

The terms used in the present application are only used to describe specific embodiments and are not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In the present application, terms such as "comprise" or "have" are intended to designate the existence of features, steps, actions, components, parts, or a combination thereof described in the specification, but one or more other features or steps It is to be understood that it does not preclude the possibility of addition or presence of, operations, components, parts, or combinations thereof.

Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs. Terms as defined in a commonly used dictionary should be interpreted as having a meaning consistent with the meaning in the context of the related technology, and should not be interpreted as an ideal or excessively formal meaning unless explicitly defined in this application. Does not.

The polyurethane-based polymer compound for a biotissue adhesive according to an embodiment of the present invention may include a first monomer of Formula 1 below and a second monomer of Formula 2 below.

[Formula 1]

[Formula 2]

In Formulas 1 and 2, R1 may be an alkene functional group having about 1 or more and 25 or less carbon atoms, R2 may be an aliphatic functional group having about 5 or more and 15 or less carbon atoms, and R3 is an aliphatic functional group having about 5 or more and 15 or less carbon atoms. I can. And a and b may each independently be an integer of about 1 or more and 3 or less.

In one embodiment, R1 may include an ethylene functional group.

In one embodiment, R2 may include one selected from the group consisting of the following Formulas 3-1 to 3-3.

[Formula 3-1]

[Chemical Formula 3-2]

[Chemical Formula 3-3]

In one embodiment, R3 may include a functional group represented by Formula 4.

[Formula 4]

In one embodiment, the polyurethane-based polymer compound for a bio-tissue adhesive may have a molecular structure represented by Formula 5 below.

[Formula 5]

In Formula 5, m and n may each independently be an integer of 1 or more and 3 or less, and a and b may each independently be an integer of about 1 or more and 3 or less.

In one embodiment, the ratio of m and n may be about 1:1 to 1:3. When the ratio of m and n is less than 1:1, the adhesive ability of the polymer compound may decrease, and when it exceeds 1:3, mechanical properties such as flexibility may decrease.

In one embodiment, the polyurethane-based polymer compound for a biotissue adhesive may have a molecular weight of about 1500 or more and 3000 g/mol or less. When the molecular weight of the polymer compound is less than 1500 g/mol, there may be a problem that the adhesive ability of the polymer is insufficient, and when it exceeds 3000 g/mol, the flexibility of the adhesive may decrease.

Hereinafter, an embodiment of a method for synthesizing the polyurethane-based polymer compound for a biological tissue adhesive will be described. However, the following synthesis method is a method for synthesizing some of the polyurethane-based polymer compounds for biotissue adhesives of the present invention, and the range of the polyurethane-based polymer compounds for biotissue adhesives of the present invention described above is limited by the following synthesis method. It should not be interpreted as.

A method for synthesizing a polyurethane-based polymer compound for a biotissue adhesive according to an embodiment of the present invention includes: a first step of synthesizing a dopamine functionalized azide compound of Formula 6; Including a second step of mixing and reacting an ethylene glycol compound of Formula 7 below, a diisocyanate compound of Formula 8, a propargyl diol of Formula 9, and the dopamine-functionalized azide compound do.

[Formula 6]

[Formula 7]

[Formula 8]

[Formula 9]

In Formulas 6 to 8, since R1, R2, and R3 are the same as R1, R2, and R3 of Formulas 1 and 2 described above, a redundant description thereof will be omitted.

In the first step, the dopamine functionalized azide compound of Formula 6 may include the compound of Formula 6-1.

[Formula 6-1]

In one embodiment, the dopamine-functionalized azide compound of Formula 6-1 is azido by reacting 3-bromo-1-propanol and sodium azide according to the following Scheme 1 Synthesizing propanol (azido propanol); Synthesizing a first intermediate compound (1) by reacting the azide propanol, glutaric anhydride, and dimethylaminopyridine (DMAP) according to Scheme 2 below; Synthesizing a second intermediate compound (2) by reacting the first intermediate compound, NHS (N-hydroxysuccinimide) and DCC (N,N'-Dicyclohexylcarbodiimide) according to Scheme 3 below; It can be synthesized by reacting the second intermediate compound, dopamine hydrochloride, and triethylamine according to Scheme 4 below to synthesize the compound of Formula 6-1.

[Scheme 1]

[Scheme 2]

[Scheme 3]

[Scheme 4]

In the second step, the ethylene glycol compound of Formula 7 may include a polyethylene glycol compound having a molecular structure of Formula 7-1 and an average molecular weight of 300 to 500 g/mol.

[Chemical Formula 7-1]

In one embodiment, the diisocyanate compound of Formula 8 may include one of the compounds of Formulas 8-1 to 8-3 below.

[Chemical Formula 8-1]

[Formula 8-2]

[Chemical Formula 8-3]

In one embodiment, the propargyl diol compound of Formula 9 is sodium hydride, diethyl-2-methylmalonate, and propargyl bromide according to the following Scheme 5. Reacting to synthesize a third intermediate compound (3); And reacting the third intermediate compound and lithium aluminum hydride according to Scheme 6 below to synthesize the propargyl diol compound of Formula 9.

[Scheme 5]

[Scheme 6]

In one embodiment, based on 1 mol of the ethylene glycol compound of Formula 7, the diisocyanate compound of Formula 8 may be mixed with at least about 5 mol, and the propargyl diol compound of Formula 9 (propargyl diol) may be mixed at least about 3 mol, and the dopamine-functionalized azide compound may be mixed at least about 1 mol. For example, based on 1 mol of the ethylene glycol compound of Formula 7, the diisocyanate compound of Formula 8 may be mixed in a ratio of about 5 mol or more and 8 mol or less, and the propargyl of Formula 9 The diol compound (propargyl diol) may be mixed at a ratio of about 3 mol or more and 5 mol or less, and the dopamine-functionalized azide compound may be mixed at a ratio of about 1 mol or more and 2 mol or less.

According to the polyurethane-based polymer compound for a biological tissue adhesive of the present invention, the catechol group of dopamine grafted on the polyurethane backbone has excellent reactivity to biological tissues containing moisture, protein, etc., so that the biological tissue of the present invention Polyurethane-based polymer compounds for adhesives may have excellent adhesion properties to living tissues. In addition, the dopamine and the polyurethane-based polymer compound for a biological tissue adhesive containing the same is not cytotoxic and thus can be applied to the human body.

Hereinafter, examples and experimental examples of the present invention will be described in detail. However, the following examples are only some embodiments of the present invention, and the scope of the present invention is not limited to the following examples.

Synthesis of DFA (dopamine functionalized azide)

[Scheme 1]

Azido propanol was synthesized according to Scheme 1 above. Specifically, 3.5 mmol of 3-bromo-1-propanol and 6.0 mmol of sodium azide were mixed with 0.5 mmol of tetrabutylammonium hydrogen sulfate at 80° C. overnight, After dissolution, the mixed solution was extracted three times using ethyl ether, and then dried over MoSO 4 to synthesize azido propanol. The residual solvent was removed through vacuum evaporation.

[Scheme 2]

An intermediate compound (1) was synthesized according to Scheme 2 above. Specifically, 30 mmol of azide propanol, 30 mmol of glutaric anhydride, and 0.3 mmol of dimethylaminopyridine (DMAP) were dissolved in 20 ml of dichloromethane overnight at room temperature, and the mixed solution was washed with 0.5M hydrochloric acid and then over MgSO4. After drying, the intermediate compound (1) described in Scheme 2 was synthesized. After drying, the residual solvent was removed through vacuum evaporation.

[Scheme 3]

Intermediate compound (2) was synthesized according to Scheme 3 above. Specifically, 15 mmol of the intermediate compound (1) and 2 mmol of NHS (N-hydroxysuccinimide) were dissolved in dichloromethane at -5°C to form a mixed solution, and then 2 mmol of DCC dissolved in 20 ml of dichloromethane was added to the mixed solution. After the addition of dropwise, the reaction was carried out overnight at room temperature to synthesize the intermediate compound (2) of Scheme 3. After the reaction, the final solution was filtered and the residual solvent was removed through vacuum evaporation.

[Scheme 4]

DFA was synthesized according to Scheme 4 above. Specifically, dopamine hydrochloride (Dopamine hydrochloride) and triethylamine (Triethylamine) were dissolved in 20 ml of methanol to form a dopamine solution. Intermediate compound (2) was added dropwise to the dopamine solution and then mixed overnight. After removing the solvent through vacuum evaporation, the residue was dissolved in 20 ml of dichloromethane, washed with 0.5M hydrochloric acid, and dried over MgSO4.

Synthesis of propargyl diol

[Scheme 5]

An intermediate compound (3) was synthesized according to Scheme 5 above. Specifically, 2.5 mmol of sodium hydride was dissolved in THF (Tetrahydrofuran) at -10°C, and 3.0 mmol of diethyl-2-methylmalonate was added dropwise thereto. The mixture was mixed for 15 minutes to form a mixed solution. Propargyl bromide was added dropwise at the same molar ratio as sodium hydride to the mixed solution, and the reaction temperature was gradually increased to room temperature, followed by stirring for 3 hours to synthesize an intermediate compound (3). After the reaction was quenched using NH4Cl, extraction was performed using ethyl ether. After drying over MgSO4, the residual solvent was removed through vacuum evaporation.

[Scheme 6]

Propargyl diol was synthesized according to Scheme 6. Specifically, 2.7 mmol of the intermediate (3) was dissolved in dichloromethane, and then 5.3 mmol of lithium aluminum hydride dissolved in dichloromethane was added dropwise at 0°C to form a mixed solution. After the mixed solution was stirred for 1 hour, a saturated NH4Cl solution was poured thereto. After washing with ethyl ether, the final solution was dried over MgSO4.

Synthesis of polyurethane-based polymer compounds

<Examples 1 to 3>

In the composition as shown in Table 1, each of the diisocyanate compounds of the following Formulas 8-1 to 8-3 was mixed and reacted with PEG 400 (polyethylene glycol 400), propargyl diol, and DFA at room temperature without a solvent, and Examples 1 to The polyurethane-based polymer compound of 3 was synthesized.

[Chemical Formula 8-1]

[Formula 8-2]

[Chemical Formula 8-3]

<Comparative Examples 1 to 3>

In the composition as described in Table 1, each of the diisocyanate compounds of Formulas 3-1 to 3-3 below was mixed and reacted with PEG 400 (polyethylene glycol 400) at room temperature without a solvent, and the polyurethane-based polymers of Comparative Examples 1 to 3 The compound was synthesized.

<Comparative Example 1>
PU_HDI
[mol] <Comparative Example 2>
PU_H12MDI
[mol] <Comparative Example 3>
PU_IPDI
[mol] <Example 1>
PU_HDI
(Dopamine)
[mol] <Example 2>
PU_H12MDI
(Dopamine)
[mol] <Example 3>
PU_IPDI
(Dopamine)
[mol] PEG 400 One One One One One One Propargyl diol >3 >3 >3 >3 >3 >3 DFA - - - One One One HDI >5 - - >5 - - H12MDI - >5 - - >5 - IPDI - - >5 - - >5

Characteristic evaluation

<Experimental Example 1>

1 and 2 are results of 1H NMR analysis and FT-IR analysis for azido propanol, intermediate compound (1), intermediate compound (2), and DFA.

1 and 2, an azido group was formed using sodium azide on bromo-propanol by a halogen substitution reaction. As shown in (A) of FIG. 6, in azido propanol, hydrogen located adjacent to the azido group appeared at 3.57 ppm (1). These results indicate that the halogen substitution reaction induces the formation of azido groups as starting materials for DFA synthesis.

After preparation of easydopropanol, the hydroxy group of azidopropanol reacted with the ring-opened glutaric anhydride by esterification to form a carboxyl group as a pendant group. As shown in (B) of Figure 1, the carboxyl groups formed by the esterification reaction appeared at 2.35 ppm (4), 4.10 ppm (3) and 5.25 ppm (5), respectively. And the azido group formed in the previous step was maintained and appeared at 3.32 ppm(1). From the data of Figure 2 (A), in the case of representative covalent bond absorptions, the hydroxy bond was at 2962.6 cm-1, the azide group was at 2100.0 cm-1, and the ester group was at 1731.0 and 1713.5. It appeared at cm-1, and at 1690.1 cm-1 for the carboxyl group.

For the reaction between the amine group of dopamine and the carboxyl group of intermediate (1), the NHS-DCC coupling reaction was induced. From (C) of FIG. 1, the NHS portion appeared at 2.59 ppm (5) and 2.74 ppm (6), and the azido group appeared at 3.29 ppm (1). As shown in (B) of Figure 2, the azido bond, an aliphatic ester bond, a cyclic compound ketone bond, and a cyclic compound amine bond, which are representative chemical bonds of the intermediate compound (2), are 2089.5 cm-1, 1816.3 & 1786.1 cm- 1, 1730.6 & 1688.1 cm-1 and 1451.7 & 1432.6 cm-1, respectively.

The amine group of the intermediate compound (2) located at the pendant site of dopamine was easily activated with the NHS-DCC coupling end stage. As shown in (D) of Figure 1, benzene ring protons (protons) appeared in the vicinity of 6ppm (4). And, the protons of the azido group were still present at 3.19 ppm(1). As shown in Figure 2 (C), azido binding and hydroxy group peaks located in the catechol group of dopamine appeared at 2101.6 cm-1 and 3182.7 cm-1. In addition, from the elemental analysis, the theoretical composition range and the experimental composition range of each synthetic compound were similar to each other as shown in Table 2. These results indicate that all chemical reactions take place according to Schemes 1 to 4 described above.

Theoretical composition Experimental composition C H O N C H O N Intermediate compound (1) 44.65 6.04 29.76 19.53 42.42 5.272 35.758 16.22 Intermediate compound (2) 47.85 5.52 29.44 17.17 44.71 5.385 35.255 14.65 DFA 55.58 7.35 21.79 15.25 47.66 7.329 31.271 13.74

3 is a 1H NMR analysis result for propargyl diol.

Referring to FIG. 3, after the previous two-step reactions, representative protons located in the propargyl group and the opposite side alkyl group appeared at 3.88 ppm (1) and 2.43 ppm (2), respectively.

Mechanical properties of polyurethane-based polymer compounds

Table 3 shows the molecular weight measurement results of the polyurethane-based polymer compounds of Examples 1 to 3.

<Molecular weight of polyurethane-based polymer compound> Molecular weight (g/mol) Example 1 1897.7 Example 2 2566.4 Example 3 2420.8

Referring to Table 3, it can be seen that the polyurethane-based polymer compounds of Examples 1 to 3 have a molecular weight applicable as a biological tissue adhesive.

Tables 4 and 5 show the results of the T-Peel test and shear test of the polyurethane-based polymer compounds of Comparative Examples 1 to 3 and Examples 1 to 3.

<T-Peel Strength of Polyurethane-Based Polymer Compounds> Max Load (N) T-Peel strength (N/mm) Comparative Example 1 2.303 0.921 Comparative Example 2 2.235 0.894 Comparative Example 3 2.284 0.913 Example 1 6.311 2.524 Example 2 6.227 2.491 Example 3 5.523 2.209

<Shear strength of polyurethane-based polymer compounds> Max Load (N) Shear strength (Mpa) Comparative Example 1 4.684 1.874 Comparative Example 2 3.829 1.532 Comparative Example 3 3.246 1.299 Example 1 12.169 4.868 Example 2 8.250 3.300 Example 3 7.044 2.818

Referring to Tables 4 and 5, it can be seen that the polyurethane-based polymer compounds of Examples 1 to 3 have significantly improved T-Peel strength and shear strength properties than the polyurethane-based polymer compounds of Comparative Examples 1 to 3. have.

From these results, it can be seen that because of the good reactivity between the catechol group of dopamine and the biological tissue containing moisture, protein, etc., dopamine plays an important role in improving adhesion properties.

Table 6 shows the contact angles measured for the polyurethane-based polymer compounds of Comparative Examples 1 to 3, DFA, and the polyurethane-based polymer compounds of Examples 1 to 3.

<Contact angle of polyurethane-based polymer compounds and dopamine> Contact angle (°) Comparative Example 1 77.8 Comparative Example 2 82.3 Comparative Example 3 81.1 DFA 51.9 Example 1 65.2 Example 2 68.6 Example 3 69.9

Referring to Table 6, DFA was found to have the highest hydrophilic properties due to the catechol group. It was found that the polyurethane-based polymer compounds of Examples 1 to 3 grafted with DFA were more hydrophilic than the polyurethane-based polymer compounds of Comparative Examples 1 to 3 not grafted to DAF. These results indicate that not only DFA was appropriately grafted to the compounds of Examples 1 to 3, but also that the grafted DFA reacted with water or amine molecules on the surface of living tissue to improve adhesion properties.

In particular, it was found that the compound of Example 1 has more hydrophilicity than the compounds of Examples 2 and 3, and for this reason, the compound of Example 1 is more than the compounds of Examples 2 and 3 as shown in Tables 4 and 5. Despite having a small molecular weight (see Table 3), it was found to have higher T-Peel strength and shear strength properties. These results indicate that HDI properly reacted with PEG without any steric hindrance and side reactions such as urea chemical reaction.

In vivo cytotoxicity test results

4 and 5 are graphs showing cell viability of DFA and polyurethane-based polymer compounds of Examples 1 to 3.

4 and 5, from the MTT-assay, mitochondrial reduction in living cells caused by the change of MTT tetrazolium (yellow) to MTT formazan (purple) was evaluated. As one of the effective evaluation tools for toxicity, cell viability was confirmed. The change in UV-vis absorption at 570 nm is directly proportional to the MTT formazan reduction. As shown in FIG. 4, DFAs exhibited an appropriate cell viability level (cell viability of 80% or more) despite their extraction amount increased from 1% to 50% of the total media volume. From these results, it can be seen that DFA grafted onto the polyurethane backbone as a biological tissue adhesive exhibits a non-cytotoxic level according to ISO 10993-5. In addition, the polymer compounds of Examples 1 to 3 showed a cell viability of 80% or more, which is regarded as a non-cytotoxic level, even though the extract amount was increased to 1 to 50%. From the MTT assay results, it can be seen that DFA and each of the polymer compounds of Examples 1 to 3 can be applied as a biological tissue adhesive without any cytotoxic effect. In the ALP-assay, changes in phosphatase derived from living cell membranes were measured using the ALP-assay kit to confirm the degree of cell activation.

As shown in FIG. 5, although their concentration was increased to 50% of the total batch volume, each of the polymer compounds of Examples 1 to 3 showed similar cellular activation. In addition, cell activation behavior between DFA and the polymer compounds of Examples 1 to 3 showed similar values for up to 20%. From these results, it is judged that DFA can improve adhesion properties without interfering with cell behavior. These results indicate that the DFA and the polymer compounds of Examples 1 to 3 are at non-toxic levels and have an appropriate appearance for the degree of cell activation according to ISO standards from in vivo cytotoxicity assay data.

In vivo animal study to evaluate short-term inflammation of polyurethane adhesives

6A and 6B are H&E staining images of an experimental rat model for the polymer compound samples of Examples 1 to 3.

6A and 6B, any tumor from the injection site of the experimental rat for 3 weeks from the initial stage to the final stage after the insertion of the polymer compound samples of Examples 1 to 3 for the experimental rat (SD-rat) tumors) and abscesses were not detected by the naked eye. The biological tissue sampling from the injection site was performed every week according to a preset time interval. In the H&E staining images, hematoxylin stained (-) charged regions of inner cells such as nucleus, whereas eosin stained (+) charged cell regions such as proteins and collagen. Colored. Macrophages surrounding living tissues produced by the reaction of the injected polymer compound samples of Examples 1 to 3 with foreign substances were colored blackish red through H&E staining. Dyed. Referring to the images of FIGS. 6A and 6B, due to the external foreign body reaction mentioned above, macrophages were generated in all biopsies of the implanted site for 1 week after surgery. However, macrophages produced in the early stages disappeared rapidly over time from 1 to 3 weeks. In addition, it was found that all the biotest sample tissues around the injection site of the experimental mice recovered well for 3 weeks. In the biotest tissues of the polymer compounds of Examples 1 to 3 containing various types of isocyanates, the amount of macrophages produced decreased sharply in the final stage (3 weeks). From these results, no chronic inflammation was observed during the 3 week transplant period, and various types of fortifying ingredients showed non-reactive behavior when injected into the human body. Therefore, it is judged that the polymer compound of Examples 1 to 3 can be applied as a medical adhesive in the surgery of soft biological tissues. In addition, immunostaining was performed to analyze the specific number of generated macrophages. In histological analysis of the presence of macrophages, CD-68 staining with antibodies that identify specific cells such as macrophages is a powerful method.

7A and 7B are immunostaining images of an experimental rat model for the polymer compound samples of Examples 1 to 3.

Referring to FIGS. 7A and 7B, macrophages were colored reddish brown color by immunostaining, and were clearly observed compared to the H&E staining results. After 3 weeks, the number of macrophages decreased, which is consistent with the data in FIGS. 6A and 6B. The number of macrophages increased during 1 week due to external foreign body reaction. However, as observed for each of the polymer compound samples of Examples 1 to 3, the macrophages almost completely disappeared after 3 weeks. These results indicate that acute inflammation is not produced by the presence of the polymer compound samples of Examples 1 to 3 beyond the initial stage after injection.

Although the above has been described with reference to preferred embodiments of the present invention, those skilled in the art will be able to variously modify and change the present invention without departing from the spirit and scope of the present invention described in the following claims. You will understand that you can.

none

Claims (13)

A polyurethane-based polymer compound for a biotissue adhesive comprising a first monomer of Formula 1 and a second monomer of Formula 2 below:
[Formula 1]

[Formula 2]

In Formulas 1 and 2, R 1 is an alkene functional group having 1 to 25 carbon atoms, R 2 is an aliphatic functional group having 5 or more and 15 or less carbon atoms, R 3 is an aliphatic functional group having 5 or more and 15 or less carbon atoms, and a and b are each It is an integer of 1 or more and 3 or less independently. The method of claim 1,
The R2 is a polyurethane-based polymer compound for a biological tissue adhesive, characterized in that it comprises any one selected from the group consisting of the following formulas 3-1 to 3-3:
[Formula 3-1]

[Chemical Formula 3-2]

[Chemical Formula 3-3]

The method of claim 1,
The R3 is a polyurethane-based polymer compound for biotissue adhesive, characterized in that it contains the functional group of Formula 4:
[Formula 4]

The method of claim 1,
Polyurethane-based polymer compound for biotissue adhesive, characterized in that it has a molecular structure of the following formula (5):
[Formula 5]

In Formula 5, m and n are each independently an integer of 1 or more and 3 or less. The method of claim 4,
The ratio of m and n is 1:1 to 1:3, characterized in that, a polyurethane-based polymer compound for biotissue adhesive. The method of claim 4,
A polyurethane-based polymer compound for biotissue adhesive, characterized in that it has a molecular weight of 1500 or more and 3000 g/mol or less. In the synthesis method of a polyurethane-based polymer compound for a biological tissue adhesive having a molecular structure of the following formula (5),
A first step of synthesizing a dopamine functionalized azide compound of Formula 6; And
Including a second step of mixing and reacting an ethylene glycol compound of Formula 7 below, a diisocyanate compound of Formula 8, a propargyl diol of Formula 9, and the dopamine-functionalized azide compound A method for synthesizing a polyurethane-based polymer compound for a biotissue adhesive, characterized in that:
[Formula 5]

[Formula 6]

[Formula 7]

[Formula 8]

[Formula 9]

In Formulas 5 to 8, R1 is an alkene functional group having 1 to 25 carbon atoms, R2 is an aliphatic functional group having 5 or more and 15 or less carbon atoms, R3 is an aliphatic functional group having 5 or more and 15 or less carbon atoms, and a and b are each Independently, it is an integer of 1 or more and 3 or less, and m and n are each independently an integer of 1 or more and 3 or less. The method of claim 7,
The dopamine-functionalized azide compound of Formula 6 is a method for synthesizing a polyurethane-based polymer compound for a biological tissue adhesive, characterized in that it comprises a compound of Formula 6-1:
[Formula 6-1]

The method of claim 7,
The first step,
Synthesizing azido propanol by reacting 3-bromo-1-propanol and sodium azide according to Scheme 1 below;
Synthesizing a first intermediate compound (1) by reacting the azide propanol, glutaric anhydride, and dimethylaminopyridine (DMAP) according to Scheme 2 below;
Synthesizing a second intermediate compound (2) by reacting the first intermediate compound, NHS (N-hydroxysuccinimide) and DCC (N,N'-Dicyclohexylcarbodiimide) according to Scheme 3 below; And
It characterized in that it comprises the step of synthesizing the compound of Formula 6-1 by reacting the second intermediate compound, dopamine hydrochloride and triethylamine according to the following Scheme 4, for biological tissue adhesive Synthesis method of polyurethane-based polymer compound:
[Scheme 1]

[Scheme 2]

[Scheme 3]

[Scheme 4]

The method of claim 7,
The diisocyanate compound of Formula 8 is a method for synthesizing a polyurethane-based polymer compound for a biotissue adhesive, characterized in that it contains any one selected from the group consisting of compounds of Formulas 8-1 to 8-3:
[Chemical Formula 8-1]

[Formula 8-2]

[Chemical Formula 8-3]

The method of claim 7,
The propargyl diol compound of Formula 9,
Synthesizing a third intermediate compound (3) by reacting sodium hydride, diethyl-2-methylmalonate, and propargyl bromide according to Scheme 5 below; And
Synthesis of a polyurethane-based polymer compound for biotissue adhesive, characterized in that it is synthesized through the step of synthesizing the propargyl diol compound of Formula 9 by reacting the third intermediate compound and lithium aluminum hydride according to Scheme 6 below. Way:
[Scheme 5]

[Scheme 6]

The method of claim 7,
In the second step,
Based on 1 mol of the ethylene glycol compound, the diisocyanate compound (diisocyanate) is mixed in a ratio of 5 mol or more and 8 mol or less, and the propargyl diol compound is mixed in a ratio of 3 mol or more and 5 mol or less. Mixed, wherein the dopamine-functionalized azide compound is mixed in a ratio of 1 mol or more and 2 mol or less, wherein the method for synthesizing a polyurethane-based polymer compound for a biological tissue adhesive. A biotissue adhesive comprising a polyurethane-based polymer compound for a biotissue adhesive according to any one of claims 1 to 6.

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