KR20120096423A - Bio-adhesive for living organism comprising poly l-3,4-dihydroxyphenylalanine and cyanoacrylate, and preparation method thereof - Google Patents

Abstract

PURPOSE: A bio-adhesive is provided to form a coating film with strong adhesion and improved physical properties, to remarkably reduce side-effect like inflammation, etc, and to have excellent profitability. CONSTITUTION: A bio-adhesive comprises poly L-3, 4-dihydroxyphenylalanine having a repeating unit in chemical formula 1, and cyano acrylate. In chemical formula 1, A is a bond or -(CH2)l-B-(CH2)m-, B is CH2 or O, l and m is respectively an integer from 1-9, the sum of l and m is 2-10, and a molecular weight of the poly L-3, 4-dihydroxyphenylalanine is 500-5,000,000. The A is a bond, C1-10 alkylene, or -(CH2)-O-(CH2)-. The A is a bond, ethylene, or -(CH2)-O-(CH2)-

Description

Bio-adhesive for living organism comprising poly L-3,4-dihydroxyphenylalanine and cyanoacrylate, and preparation method according to the present invention, including bio-adhesive for living organism comprising poly L-3,4-dihydroxyphenylalanine and cyanoacrylate.

The present invention relates to a bioadhesive capable of forming a coating film having strong bonding strength and improved physical properties with an adherend and a method of manufacturing the same.

In the medical field, particularly in the field of surgical treatment, when the skin is incised by a surgical operation or the like, the suture of the finally incised skin should proceed.

At the time of suture, non-degradable or biodegradable sutures are generally used. The larger the incision site, the longer the suture length and the number of suture units are increased. In addition, after the suture is inconvenient to change the bandage often after covering with a bandage, etc., there is also a problem that the area where the suture passed by remains as a scar. In particular, for many exposed areas such as the face, hands, and legs, patients are reluctant to use the suture, and there are various problems such as requiring a second surgery to remove scars.

In order to replace the suture using the suture, a method of using a bioadhesive is known. Bioadhesive means an adhesive used for bonding between biological tissues such as skin, blood vessels or bones, or an artificial material and biological tissues, and is known to be used instead of sutures in some large hospitals.

Examples of bioadhesives include cyanoacrylate adhesives, fibrin glues, gelatin glues and urethane adhesives (Korean Patent Registration No. 10-0328733, Korean Patent Registration No. 10-0450142), among which cyanoacrylate is used. It is known that the polymerization is initiated from the monomer by the nucleophilic attack by the OH- of the moisture on the skin surface or in the air and proceeds to the chain extension step and the end of the chain extension (D. Kotzev, V. Kotzev, International Journal of Adhesion and Adhesives, Volume 12, Issue 3, July 1992, Pages 150-157; Pratik J. Mankidy, Ramakrishnan Rajagopalan, Henry C. Foley, Polymer, Volume 49, Issue 9, 29 April 2008, Pages 2235-2242). On the other hand, since the polymerization termination reaction is very fast, the industrial adhesive using the same is called an instant adhesive.

On the other hand, cyanoacrylates are known to have little toxicity in the body depending on the type of alkyl in the molecule (Gabriela Ciapetti, Susanna Stea, Elisabetta Cenni, Alessandra Sudanese, Daniela Marraro, Aldo Toni, Arturo Pizzoferrato, Biomaterials, Volume) 15, Issue 1, 1994, Pages 63-67; CE Evans, GC Lees, IA Trail, The Journal of Hand Surgery: Journal of the British Society for Surgery of the Hand, Volume 24, Issue 6, December 1999, Pages 658- 661) On the basis of this, attempts and related products have been reported for application to areas such as skin, nails or toenails, and sutures.

Currently commercially available Octyl 2-cyanoacrylate (trade name: Dermabond) and N-Butyl 2-cyanoacrylate (trade name: Histoacryl) are both finished products imported from the United States or Germany. The above products have rapid reaction with water and polyalkylcyanoacrylate after polymerization has very hard properties, so it can be applied to biological tissues, but due to differences in physical properties in flexible areas such as skin after the procedure In many cases, complaints of discomfort and rejection are reported, and incontinence or whitening is often reported due to the weakening of the impact strength.

In order to solve the above problems, there have been attempts to change the properties of cyanoacrylate. For example, when cyanoacrylate is mixed with polyethylene glycol (PEG) or a low molecular weight ether-based adhesive agent or ester plasticizer, physical properties change. There have been reports of poly (DL-lactide) (PDLA) or poly (lactide-co-glycolide) (PLGA). 0523662; Korean Patent Registration No. 10-0810836; Korean Patent Registration No. 10-0827264).

In addition, studies have been reported to control physical properties by chemically modifying cyanoacrylate (Korean Patent Publication No. 10-2009-0077759). However, low molecular weight plasticizers and PEG may impart plasticity, but they may elute from the polymer and affect the body. PDLA and PLGA have relatively hard physical properties but are insufficient to impart strong adhesion. have. In addition, chemical denaturation affects the polymerization reaction and biocompatibility, so it can only be used in extremely limited industrial adhesives. Due to the above problems, there is a need for the development of biocompatible cyanoacrylates with improved adhesion without the use of low molecular weight plasticizers and the like without the use of artificially reactive adhesives.

Accordingly, the present inventors have studied the bioadhesives in which the above problems are solved, and the bioadhesives including poly L-3,4-dihydroxyphenylalanine and cyanoacrylate have strong binding power and improved physical properties with the adherend. It is confirmed that it is possible to form a coating film, and also to control the viscosity of the adhesive, thereby inhibiting the penetration of the adhesive component into the wound, thereby significantly reducing side effects such as inflammation and providing a bioadhesive that can enhance economic efficiency. Thus, the present invention has been completed.

The present invention is to provide a bioadhesive capable of forming a coating film having a strong bond with the adherend and improved physical properties.

In another aspect, the present invention is to provide a method for producing the biological adhesive.

In order to solve the above problems, the present invention provides a bioadhesive comprising poly L-3,4-dihydroxyphenylalanine and cyanoacrylate having a repeating unit represented by the following formula (1).

[Formula 1]

Where

A is a bond or - _{a, - (CH 2) l -B-} (CH 2) m

B is CH ₂ or O,

L and m are each an integer of 1 to 9,

The sum of l and m is 2 to 10, and

The molecular weight of the poly L-3,4-dihydroxyphenylalanine is 500 to 5,000,000.

Preferably, A is a bond, C _{1 -10} alkylene, or _{- (CH 2) -O- (CH} 2) -, more preferably wherein A is a bond, an ethylene (-CH ₂ -CH ₂ - ), or _{- (CH 2) -O- (CH} 2) - a.

The basic structure of the repeating unit represented by Formula 1 is 3,4-dihydroxyphenylalanine (DOPA), in particular L-3,4-dihydroxyphenylalanine (L-DOPA), and has a structure of a modified amino acid of phenylalanine (Phenylalanine), A substance found in mussels that is strongly bound to rock. Rejection or side effects are not known in the body, and it is known to exert strong adhesion in the presence of moisture. In the present invention, a double bond is introduced into the compound to be used as a monomer of a polymer, and the double bond can be introduced by a method of introducing an epoxy alkene.

As used herein, the term 'cyanoacrylate' refers to an acrylate including a cyano group. For example, methyl 2-cyanoacrylate, ethyl 2-cyanoacrylate (Ethyl 2-cyanoacrylate, Butyl 2-cyanoacrylate, Isohexyl 2-cyanoacrylate, Allyl 2-cyanoacrylate, Octyl 2 -Octyl 2-cyanoacrylate, Ethoxyethyl 2-cyanoacrylate, Isobutyl 2-cyanoacrylate or hexyl 2-cyanoacrylate ( Hexyl 2-cyanoacrylate).

The composition comprising the poly L-3,4-dihydroxyphenylalanine and cyanoacrylate can form a coating film having a strong binding strength and improved physical properties with the adherend, it can be usefully used as a bioadhesive.

As used herein, the term 'bioadhesive' refers to an adhesive used for binding between biological tissues such as skin, blood vessels or bones, or for bonding an artificial material to biological tissues. The bioadhesive according to the present invention is instantaneously bonded, and at this time, no heat or harmful substances are generated, and rejection does not occur. Therefore, the bioadhesive according to the present invention can be usefully used in the medical field.

In the bioadhesive according to the invention, the cyanoacrylate is preferably present at 0.01-80% of the mass of poly L-3,4-dihydroxyphenylalanine.

In addition, the present invention comprises the steps of preparing a compound represented by the following formula (4) by reacting the compound represented by the formula (2) and the compound represented by the formula (3) as shown in the following reaction scheme (step 1); And

It provides a step (step 2) of preparing a poly L-3,4-dihydroxyphenylalanine by polymerizing the compound represented by the formula (4).

[Reaction Scheme 1]

The definition of A in the above scheme is as defined above.

Step 1 is a reaction for introducing a double bond into the compound represented by the formula (2), the step of reacting the amino group of the compound represented by the formula (2) with the epoxy group of the compound represented by the formula (3).

In the reaction, the mass ratio of the compound represented by Formula 2 and the compound represented by Formula 3 is preferably 1: 1 to 1:10.

The reaction may be performed by stirring at room temperature for about 5 hours, whereby a double bond may be introduced into the compound represented by Formula 2.

Step 2 is a step of preparing poly L-3,4-dihydroxyphenylalanine by polymerizing using a double bond of the compound represented by the formula (4).

The reaction may be carried out by a photopolymerization method or a reaction by a radical initiator at room temperature and heating conditions. In the method by photopolymerization, 2- (dimethylamino) ethyl methacrylate (DEMA), ethyl 4-dimethylaminobenzoate together with camphorquinon (CQ), an initiator for photopolymerization, (ethyl 4-dimethylaminobenzoate; E4), diphenyliodonium hexafluorophosphate (diphenyliodonium hexafluorophosphate; mixed with one or more photosensitizers selected from DPIHP) can be irradiated with ultraviolet light of 400 nm or less for about 10 seconds have. In addition, in reactions with radical initiators under heating and room temperature conditions, benzoyl peroxide, acetyl peroxide, dilauryl peroxide, di-tert-butyl peroxide (di-tert) -butyl peroxide, cumyl hydroperoxide, hydrogen peroxide, potassium persulfate, ammonium persulfate or azobisisobutyronitrile (AIBN) Radical initiators such as can be used. At this time, a solvent such as hydrochloric acid, sulfuric acid, nitric acid, acetic acid, formic acid or p-toluenesulfonate may be added.

The bioadhesive comprising poly L-3,4-dihydroxyphenylalanine and cyanoacrylate according to the present invention can form a coating film having strong binding strength and improved physical properties with the adherend, and also easy to control viscosity. By inhibiting the penetration of the adhesive component into the wound can not only significantly reduce side effects such as inflammation, but also improve the economics, it can be useful as a bioadhesive in the medical field.

Figure 1 shows the bonding strength of the bioadhesive according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for describing the present invention more specifically, and the scope of the present invention is not limited by these examples.

Example  One

L-3,4-dihydroxyphenylalanine (L-DOPA) to 3,4-epoxy-1-butene (3,4-Epoxy-1-butene) in a mass ratio of 1: 2 After mixing, the mixture was stirred at room temperature for about 5 hours.

To the reaction solution, benzoyl peroxide was added 0.3% of the total weight ratio, and the mixture was stirred at 70 ° C. for 20 minutes, and the viscosity increase was observed in real time. When the viscosity reached 70 mPa? S, the reaction was stopped and a viscous transparent solution was recovered.

The solution was mixed by adding 25% by mass to a butyl 2-cyanoacrylate solution and stirred vigorously for 1 hour. The mixed solution with viscous and slightly turbid properties was stored frozen after filling with nitrogen.

Example  2

L-3,4-dihydroxyphenylalanine (L-DOPA) to 1,2-epoxy-5-hexene (1,2-epoxy-4-hexene) in a mass ratio of 1: 3 After mixing, the mixture was stirred at room temperature for about 5 hours.

To the reaction solution, benzoyl peroxide was added 0.3% of the total weight ratio, and the mixture was stirred at 70 ° C. for 20 minutes, and the viscosity increase was observed in real time. When the viscosity reached 50 mPa? S, the reaction was stopped and a viscous transparent solution was recovered.

The solution was mixed by adding 10% by mass to allyl 2-cyanoacrylate solution and stirred vigorously for 1 hour. The mixed solution with viscous and slightly turbid properties was stored frozen after filling with nitrogen.

Example  3

After mixing allyl glycidyl ether with L-3,4-dihydroxyphenylalanine (L-DOPA) in a mass ratio of 1: 3, at room temperature for about 5 hours. Stirred.

To the reaction solution, benzoyl peroxide was added 0.3% of the total weight ratio, and the mixture was stirred at 70 ° C. for 20 minutes, and the viscosity increase was observed in real time. When the viscosity reached 40 mPa? S, the reaction was stopped and a viscous transparent solution was recovered.

The solution was mixed by adding 20% by mass to ethyl 2-cyanoacrylate solution and stirred vigorously for 1 hour. The mixed solution with viscous and slightly turbid properties was stored frozen after filling with nitrogen.

Experimental Example  1: Bond strength measurement

The bond strength, which is a main characteristic of the bioadhesive, was measured as follows. Specifically, 1 g of the viscous adhesive prepared in Examples 1 to 3 was applied to bovine skin (10 mm × 10 mm), respectively. Another bovine skin was again covered on the coating layer and allowed to dry naturally for 24 hours, and then pulled at a speed of 1 mm / min using a mechanical strength meter (Instron model 4467, Canton, MA, USA). Was measured as the bonding strength, and the results are shown in FIG.

As shown in FIG. 1, the 3,4-epoxy-1-butene of Example 1 was substituted with 1,2-epoxy-5-hexene (1,2- of Example 2). It was expected to have higher reactivity with lower molecular weight than epoxy-4-hexene), but there was a significant difference between Examples 1 and 2 due to the difference in self-adhesive butyl 2-cyanoacrylate and allyl 2-cyanoacrylate of each example. No difference was seen. However, despite the use of ethyl 2-cyanoacrylate, which is relatively weak in Example 3, due to the molecular structure of allylglycidyl ether, a harder adhesive was obtained than other examples. It could be confirmed that the bond strength was shown.

Experimental Example  2: cytotoxicity measurement

In order to confirm that the bioadhesives exhibit cytotoxicity, the following experiment was performed. Specifically, 10,000 fibroblasts were inoculated in a 24 well plate and after 24 hours, the adhesives of Examples 1 to 3 were each added 30 ul. Dermabond (derma bond), a bioadhesive used as a control group, was added in the same manner, and 1.5 ml of the medium was incubated for 24 hours, and then absorbance of 590 nm was measured by MTT assay. The results are shown in Table 1 below.

Biometric adhesive Dermabond Example 1 Example 2 Example 3 Cell survival rate (%) 52 ± 4 31 ± 5 42 ± 3 38 ± 3

As shown in Table 1, in particular, it was confirmed that the toxicity of Example 2 is low.

Claims (10)

A bioadhesive comprising a poly L-3,4-dihydroxyphenylalanine having a repeating unit represented by the following Formula 1, and cyanoacrylate:
[Formula 1]

In this formula,
A is a bond or - _{a, - (CH 2) l -B-} (CH 2) m
B is CH ₂ or O,
L and m are each an integer of 1 to 9,
The sum of l and m is 2 to 10, and
The molecular weight of the poly L-3,4-dihydroxyphenylalanine is 500 to 5,000,000.
2. The method of claim 1, wherein A is a bond, C _{1 -10} alkylene, or _{- (CH 2) -O- (CH} 2) - , characterized in that bio-adhesive.
The bioadhesive of claim 2, wherein A is a bond, ethylene, or — (CH ₂ ) —O— (CH ₂ ) —.
The method of claim 1, wherein the cyanoacrylate is methyl 2-cyanoacrylate, ethyl 2-cyanoacrylate, butyl 2-cyanoacrylate, isohexyl 2-cyanoacrylate, allyl 2-cyano At least one selected from the group consisting of acrylate, octyl 2-cyanoacrylate, ethoxy 2-cyanoacrylate, isobutyl 2-cyanoacrylate and hexyl 2-cyanoacrylate Biored chopsticks.
The bioadhesive of claim 1, wherein the cyanoacrylate is present at 0.01 to 80% of the mass of the poly L-3,4-dihydroxyphenylalanine.
Preparing a compound represented by the following Chemical Formula 4 by reacting the compound represented by the following Chemical Formula 2 with the compound represented by the following Chemical Formula 3 (step 1); And
To prepare a poly L-3,4-dihydroxyphenylalanine represented by the following formula (1) by polymerizing a compound represented by the formula (4), poly L-3,4-dihydroxyphenylalanine Manufacturing Method:
[Formula 1]

(2)

(3)

[Chemical Formula 4]

In Chemical Formulas 1, 3, and 4, the definitions of A and n are the same as those of claim 1.
The method of claim 6, wherein in step 1, the mass ratio of the compound represented by Chemical Formula 2 and the compound represented by Chemical Formula 3 is 1: 1 to 1:10.
The method of claim 6, wherein the reaction of Step 2 is any one selected from camphorquinone and 2- (dimethylamino) ethyl methacrylate, ethyl 4-dimethylaminobenzoate and diphenyliodonium hexafluorophosphate. Method of producing a UV light in the presence of the above photosensitizers.
The method of claim 6, wherein the reaction of Step 2 is performed by: benzoyl peroxide, acetyl peroxide, diauyl peroxide, di-tert-butyl peroxide, cumyl hydroperoxide, hydrogen peroxide, potassium persulfate, ammonium peroxide A process characterized in that it is carried out in the presence of a radical initiator selected from the group consisting of sulfate and azobisisobutyronitrile.
10. The method according to claim 8 or 9, wherein the reaction of step 2 is added with a solvent selected from the group consisting of hydrochloric acid, sulfuric acid, nitric acid, acetic acid, formic acid and p-toluenesulfonate.

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