KR20130133692A - Method of manufacturing crosslinked hydrogel for drug delivery system - Google Patents

Abstract

The present invention relates to a medical material, more specifically, to a medical material which contains a catechol-based functional group with excellent adhesive force and coating ability as an active ingredient to be provided as an effective drug carrier; has excellent osmotic pressure absorption, waterproof, and moisture permeable functions when the material touches a wound; prevents the invasion of bacteria, virus, and foreign materials from the outside; promotes wound healing by providing proper humid environment to the wound; minimizes the scar after healing; and prevents the synechia of organs. [Reference numerals] (AA) Wash first curved tempered glass;(BB) Example 2 [After 12hr (G")];(CC) Example 2 [Initial Time (G')];(DD) Example 2 [Initial Time (G")];(EE) Control group [After 12hr (G')];(FF) Control group [After 12hr (G")];(GG) Control group [Initial Time (G')];(HH) End

Description

Cross-linked hydrogel for drug delivery and method for producing the hydrogel {Method of Manufacturing Crosslinked Hydrogel for Drug Delivery system}

The present invention relates to a matrix material for drug delivery. More specifically, the present invention relates to a crosslinked hydrogel that can be used in a medical material including a biomaterial conjugated with catechol having excellent adhesion.

Drug Delivery System (DDS) is a dosage form designed to efficiently deliver the required amount of drugs by minimizing the side effects of existing drugs and maximizing efficacy and effectiveness. Biocompatible polymers for injectable biocompatible and biodegradable hydrogels have been developed for efficient drug delivery of macromolecules such as proteins and genes.

Such polymers are carried out through chemical and physical crosslinking for stability and stability in vivo, but some crosslinking agents may be used for chemical bonding. The material after such crosslinking may cause side effects such as inflammation due to increased toxicity and residual material. In the case of hydrogel, variables such as effective drug loading rate and persistence are important.

In the case of hydrogels, when tissue damage occurs after drug delivery, surgery, or inflammation, a natural wound healing mechanism proceeds. When excessive fibrous tissue is generated in this process, the surrounding tissues and abnormal tissues are abnormal. It can also be used as an anti-adhesion material to prevent bonding. Adhesions after abdominal surgery may cause sequelae such as intestinal dysfunction, intestinal obstruction, and chronic pain. In particular, adhesions after gynecological surgery cause infertility.

In obstetrics and gynecology, the adhesion to the endometrium occurs in 20-50% after curettage or inhalation of incomplete abortion, stillbirth, recurrent miscarriage, etc. A lot of research is being done for this purpose, and a method of using a physical barrier has recently been developed and used.

Physical barriers that are effective in preventing adhesion require the flexibility to encapsulate the wound and isolate it from surrounding tissues while serving as a tissue or physical barrier while the wound heals. In addition, after regeneration of damaged tissues, the body must decompose itself, be absorbed and released, and physical barriers or products decomposed therefrom must not cause toxic or inflammatory reactions in the body. Biodegradable polymers are used as materials for such anti-adhesion agents.

Biodegradable polymers are those that are chemically decomposed in the body and gradually lose their form and weight.As a natural biodegradable polymer, collagen, fibronectin, gelatin, chitosan ), Alginic acid, hyaluronic acid, etc. are processed and the synthetic biodegradable polymer is poly (lactic acid) (PLA), poly (glycolic acid) (PGA), poly (D, L -latide-co-glycolide (PLGA) and its similar copolymers with poly (ε-caprolactone) (PCL), polyanhydrides and polyesters.

Hyaluronic acid is a non-sulfated glycosaminoglycan widely distributed throughout connective tissue, epithelial tissue and neural tissue. Hyaluronic acid is composed of D-glucuronic acid and D-N-acetylglucosamine, and is a disaccharide polymer in which β-1,4 and β-1,3 glucosidic bonds are alternately bonded. In addition, the Pluronic F127 anti-adhesion agent has the advantage of ease of use in the form of a sol at room temperature and gel form at body temperature, but has to be used only on a dry, bleed-free surface.

In addition, the anti-adhesion agent made of PLA or PGA, which is a synthetic biodegradable polymer, has a long biodegradation period of 4 weeks or more and high mechanical strength, but because it is a hydrophobic material, it does not adhere well to the surface of biological tissue due to its low water absorption. There is a problem that the hydrolyzed to release the acidic decomposition products may cause an inflammatory reaction that causes the adhesion.

(Patent Document 0001) Domestic Publication No. 10-2008-0104228

(Non-Patent Document 0001)

Soft Matter, Yuhan Lee, Hyun Jung Chung, Sangho Yeo, Cheol-Hee Ahn, Heashin Lee, Phillip B. Messersmith and Tae Gwan Park, 977-983

(Non-Patent Document 0002)

Chem. Commun., Yeon Jeong Oh, a Il Hwan Cho, b Haeshin Lee, c Ki-Jung Park, d Hyukjin Lee and Sung Young Park, 2012, 48, 11895-11897

The present invention has been made to solve the above problems, according to one embodiment of the present invention, used as an injectable carrier of protein and stem cells, and also in contact with the wound, excellent osmotic pressure absorption, waterproofing, An object of the present invention is to provide a medical material having moisture permeability.

In addition, according to an embodiment of the present invention, it is possible to prevent the invasion of bacteria, bacteria, foreign substances, etc. from the outside, and to provide a suitable wet environment for wounds to promote wound healing such as wounds, burns, cuts, etc. The purpose is to provide.

In addition, according to an embodiment of the present invention, it is an object of the present invention to provide a medical material that can minimize the scar remaining after wound recovery and prevent the adhesion phenomenon of the organ.

In addition, according to an embodiment of the present invention, to provide a medical material that can be variously applied in all medical fields, such as otolaryngology, ophthalmology, dentistry, neurosurgery having adhesion, biocompatibility and water resistance in the aqueous solution.

A first object of the present invention is a crosslinked hydrogel comprising a compound having a catechol-based functional group as an active ingredient, i) formed by crosslinking of compounds having a catechol-based functional group, or ii) a compound having a catechol-based functional group Is formed by conjugation to a biocompatible polymer, or (iii) is formed by crosslinking with a bio / synthetic polymer having a catechol-based functional group and a bio / synthetic polymer having a -NH2 or -SH group at its end, or iii) a catechol-based functional group. And it can be achieved as a crosslinked hydrogel, characterized in that formed by crosslinking of the bio / synthetic polymer having a -NH2 or -SH group and the bio / synthetic polymer having a -NH2 or -SH group at the end.

In addition, compounds having a catechol-based functional group are 2-chloro-3 ', 4'-dihydroxyacetophenone, dopa, dopamine, caffeic acid, garlic acid, chlorogenic acid, (-)-epicatechin, (-)- Epicatechin gallate, melanin, tannin, flavonoid, cyanidin, dihydroquecetin, oron, dihydromyrithinin, delphinidine, mycetin, quercetin, (-)-epicalatechin-3-gallate, (-)-epigal Rocatechin, (-)-epigallocatechin gallate, catechin and luteolin may be characterized in that any one or more selected.

In addition, the biocompatible polymer is hyaluronic acid (hyaluronic acid), chitosan, chitosan derivatives (chitalac), collagen (collagen), fibronectin, collagen (collagen), gelatin (gelatin), alginic acid , Pluronic, poly (lacticacid) (PLA), poly (glycolicacid) (PGA), poly (D, L-latide-co-glycolide) (PLGA), poly (ε-caprolactone) (PCL), Polyanhydride, polyester (polyester), lysine (Lysine), Polyethyleneimine (PEI) and glutathione (GSH) may be characterized in that any one or more selected.

In addition, the bio / synthetic polymer having a catechol-based functional group may be any one or more of the compounds represented by the following Chemical Formulas 1 to 4.

[Formula 1]

In Formula 1,

x + y is an integer from 200 to 6,000, y is an integer from 1 to 50,

(2)

In Formula 2,

A is SH, NH2 functional group, n is 1-3, x + y is an integer of 200 to 6,000, y and z are an integer of 1 to 50,

(3)

In Formula 3,

Z is a compound having a catechol-based functional group, n + m is an integer of 10 to 700,

n is an integer from 0 to 699, m is an integer from 1 to 700, x is from 1 to 700

Is an integer of

[Chemical Formula 4]

In Formula 4, Z is a compound having a catechol-based functional group,

n + m is an integer from 10 to 700, n and m are each an integer from 1 to 700,

Bio-synthetic polymer having the -NH2 or -SH group is chitosan, chitosan derivative (chitalac), amine pluronic, catechol amine, thiol pluronic, amine hyaluronic acid, thiol hyaluronic acid, lysine (Lysine) , Polyethyleneimine (PEI) and glutathione (GSH) may be characterized in that any one or more selected.

In addition, the bio-synthetic polymer having an —NH 2 or —SH group may be any one or more of the compounds represented by the following Chemical Formulas 6 to 8.

[Chemical Formula 5]

In Formula 5, n + m is an integer of 10 to 700, n is an integer of 1 to 700,

[Chemical Formula 6]

In Formula 6, x is an integer of 1 to 12, n is an integer of 78 to 100, m is an integer of 30 to 65,

[Formula 7]

In Formula 7, x is an integer of 1 to 12, n is an integer of 78 to 100, m is an integer of 30 to 65.

In addition, the bio / synthetic polymer having a catechol-based functional group may be formed by conjugating the compound having the catechol-based functional group with the bio / synthetic polymer.

And, in the cross-linked hydrogel of iii), the compound having a catechol-based functional group is included in 0.1 to 99.9% by weight, the bio / synthetic polymer having NH2 or SH functional group is contained in 0.1 to 99.9% by weight You can do

In addition, the bio / synthetic polymer having a catechol-based functional group and the NH 2 or —SH group may be formed by conjugating the compound having the catechol-based functional group with the bio / synthetic polymer having the NH 2 or —SH group.

In the crosslinked hydrogel of iii), the catechol-based functional group and the compound having NH 2 or SH functional group are included in an amount of 0.1 to 98 wt%, and the bio / synthetic polymer having an NH 2 or -SH group is 0.1 to 98 wt%. It is included as, the bio-synthetic polymer having a -NH2 or -SH group at the terminal may be characterized in that it comprises 1 to 50% by weight.

In addition, the crosslinked hydrogel according to the first object of the present invention may be characterized in that it is formed at 20 to 80 ℃.

A second object of the present invention is a method for producing a crosslinked hydrogel comprising a compound having a catechol-based functional group as an active ingredient, comprising a solution in which a bio / synthetic polymer having a catechol-based functional group is dissolved in a solvent, and -NH 2 at the terminal. Or step 1 of mixing a solution of a bio-synthetic polymer having a -SH group in a solvent; maintaining at a pH of at least 8.5; And it can be achieved as a method for producing a crosslinked hydrogel, characterized in that it comprises a step 3 to remove impurities by centrifugation.

In addition, the bio / synthetic polymer having a catechol-based functional group is added to the solution in which the bio / synthetic polymer is dissolved in a solvent, and then reacted in a nitrogen atmosphere under an acidic condition of pH 5 or below by adding the compound having the catechol-based functional group thereto. Dialysis under acidic conditions of pH 4 or less may be characterized in that the catechol group is formed by conjugation to the bio-synthetic polymer.

This step 2 may be characterized in that it is maintained for 1 to 24 hours at a temperature of 20 to 80 ℃.

In addition, step 3 may be characterized in that it further comprises the step of producing a freeze-dried scaffold.

Therefore, according to one embodiment of the present invention as described, it can be used as a matrix for differentiation of stem cells while maximizing the bioavailability of drugs such as proteins using an injectable hydrogel. In addition, it has excellent osmotic pressure, absorption capacity, water resistance, moisture permeability by contacting the wound area, prevents the invasion of bacteria, bacteria, foreign substances, etc. from the outside, and provides an appropriate moist environment to the wound, In addition to promoting wound healing, scars remaining after wound healing can be minimized and organ adhesion can be prevented.

In addition, according to one embodiment of the present invention, it is excellent in adhesion and coating power, can be used as an anti-adhesion agent, a wound dressing agent, an anti-wrinkle agent or a medical adhesive. There is an advantage that it can be applied in a variety of medical fields such as ophthalmology, dentistry, neurosurgery.

1 is a graph showing the strength measurement results of catechol hyaluronic acid (HA-C) and chitosan derivatives (chitalac) crosslinked hydrogel according to an embodiment of the present invention.
Figure 2 is a graph showing the results of the strength measurement of catechol hyaluronic acid (HA-C) and chitosan derivatives (chitalac) crosslinked hydrogel according to an embodiment of the present invention over time.
Figure 3 shows the swelling degree (%) measurement results of the catechol hyaluronic acid (HA-C) and chitosan derivative (chitalac) crosslinked hydrogel according to an embodiment of the present invention over time.
Figure 4 is a graph showing the results of strength measurement after swelling the catechol hyaluronic acid (HA-C) and chitosan derivative (chitalac) crosslinked hydrogel according to an embodiment of the present invention over time.
Figure 5 is a graph showing the results of the strength measurement of catechol hyaluronic acid and amine pluronic crosslinked hydrogel according to an embodiment of the present invention.
Figure 6 is a graph showing the results of the strength measurement of catechol hyaluronic acid and thiol pluronic crosslinked hydrogel according to an embodiment of the present invention.
Figure 7 is a graph showing the results of the strength measurement of catechol chitalac and amine pluronic crosslinked hydrogel according to an embodiment of the present invention.
8 is a graph showing the results of measuring the strength of the catechol chitalac and thiol pluronic crosslinked hydrogel according to an embodiment of the present invention.
Figure 9 is a graph showing the strength measurement results of the crosslinked hydrogels according to one embodiment of the present invention.
Figure 10 is a photograph measuring the particle size of the cross-linked hydrogels for drug delivery according to one embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. However, the detailed description of known functions and configurations incorporated herein will be omitted when it may unnecessarily obscure the subject matter of the present invention.

The same reference numerals are used for portions having similar functions and functions throughout the drawings. Throughout the specification, when a part is connected to another part, this includes not only the case where it is directly connected, but also the case where it is indirectly connected with another element in between. In addition, the inclusion of an element does not exclude other elements, but may include other elements, unless specifically stated otherwise.

Hereinafter will be described in detail the configuration, preparation method and experimental example of the drug delivery cross-linked hydrogel according to an embodiment of the present invention.

According to an embodiment of the present invention, the crosslinked hydrogel includes a compound having a catechol-based functional group as an active ingredient, and iii) crosslinking between a compound having a catechol-based functional group and a compound having functional groups (SH and NH 2). Or ii) crosslinking a biopolymer having a catechol-based functional group and a SH or NH 2 functional group with a compound having functional groups (SH and NH 2).

Hereinafter, the crosslinked hydrogel of the present invention will be described in detail as follows.

According to an embodiment of the present invention, a crosslinked hydrogel may be formed by crosslinking of compounds having a catechol-based functional group.

The compound having a catechol-based functional group is a compound having a catechol group having adhesion, dopa, dopamine, caffeic acid, garlic acid, chlorogenic acid, (-)-epicatechin, (-)-epicatechin gallate, melanin, tannin, Flavonoids, cyanidins, dihydro quercetin, oron, dihydromyricetin, delphinidin, myrcetin, quercetin, (-)-epicalatechin-3-gallate, catechin, luteolin and the like can be used.

 According to another embodiment of the present invention, a compound having a catechol-based functional group may be conjugated to a biocompatible polymer to form a crosslinked product.

Since the compound having the catechol-based functional group is the same as described above, a detailed description thereof will be omitted.

The compound having a catechol-based functional group is preferably included in 0.1 to 99.9% by weight, more preferably 1 to 90% by weight in the crosslinked product. When the content is included in the above range, it is more preferable for the catechol-based crosslinking.

The biocompatible polymer is a polymer that is chemically decomposed in the body, and can be used without limitation as long as it is a conventional biocompatible polymer used in the art. For example, as a natural polymer, hyaluronic acid, chitosan, chitosan derivatives ( Chitalac), polymer having -NH2 or -SH group at the end, collagen, collagen, fibronectin, gelatin, alginic acid, albumin (albumin), lysine, glutathione ( GSH) may be used, and synthetic polymers include poly (lactic acid) (PLA), poly (glycolic acid) (PGA), poly (D, L-latide-co-glycolide) (PLGA), poly (ε- caprolactone (PCL), polyanhydride, polyester, polyethyleneimine (PEI) and the like can be used. At this time, the hyaluronic acid has a molecular weight of 100 ~ 2,000kDa, chitosan is preferably a molecular weight of 1 ~ 220kDa.

The biocompatible polymer is preferably included in the crosslinked material in an amount of 0.1 to 99.9% by weight, more preferably 10 to 99% by weight. When the content is included in the above range, it is more preferable for the catechol-based crosslinking.

According to another embodiment of the present invention, a compound having a catechol-based functional group is conjugated with a bio / synthetic polymer to form a bio / synthetic polymer having a catechol-based functional group, and a biological having a -NH 2 or -SH group at the terminal thereof. The crosslinked hydrogel may be formed by crosslinking with the synthetic polymer or the biocompatible polymer.

The compound having a catechol-based functional group is the same as described above, it is preferably included in the cross-linked hydrogel 0.1 to 98% by weight, more preferably 1 to 70% by weight. When the content is included in the above range, it is more preferable for the catechol-based crosslinking.

The biocompatible polymer is also the same as described above, it is preferably included in the crosslinked hydrogel 0.1 to 98% by weight, more preferably 1 to 50% by weight. If the content is included in the above range is better in crosslinking.

The compound having a catechol-based functional group is conjugated to a biocompatible polymer. Specifically, the bio / synthetic polymer is dissolved in a solvent (a solution of pH 7.4), and then the compound having a catechol-based functional group is added to the solution to pH. After the reaction at room temperature in a nitrogen atmosphere at an acidic condition of 5 or less, and then dialysis under an acidic condition of pH 4 or less, a final compound in which a catechol group is conjugated to a biocompatible polymer can be obtained.

The bio-synthetic polymer having a catechol-based functional group formed by conjugation of the compound having a catechol-based functional group and the bio / synthetic polymer may be a compound represented by the following Chemical Formulas 1 to 4.

[Formula 1]

In Formula 1,

x + y is an integer from 200 to 6,000 and y is an integer from 1 to 100.

(2)

In Formula 2,

A is SH, NH2 functional group, n is 1-3

x + y is an integer from 200 to 6,000, y and z are integers from 1 to 50,

(3)

In Formula 3,

Z is a compound having a catechol-based functional group,

n + m is 10 to 700,

n is an integer of 0-699, m is an integer of 1-700, x is an integer of 0-700.

[Chemical Formula 4]

In Chemical Formula 4,

Z is a compound having a catechol-based functional group,

n is an integer of 1 to 100 and m is an integer of 1 to 100, respectively.

In Chemical Formulas 3 and 4, Z is a compound having a catechol-based functional group, specifically, dopa, dopamine, caffeic acid, garlic acid, chlorogenic acid, (-)-epicatechin, (-)-epicatechin gallate, melanin, tannin , Flavonoids, cyanidins, dihydro quercetin, oron, dihihydromyricetin, delphinidin, mycetin, quercetin, (-)-epicarocatechin-3-gallate, (-)-epigallocatechin, (-) Epigallocatechin gallate, catechin or luteolin.

A biocompatible polymer having a catechol-based functional group formed by conjugating a compound having a catechol-based functional group with a biocompatible polymer as described above is then crosslinked with a bio / synthetic polymer having a -NH 2 or -SH group or a biocompatible polymer.

Bio-synthetic polymer having a -NH2 or -SH group at the terminal is chitosan, chitosan derivatives (chitalac), amine pluronic, catechol amine, thiol pluronic, amine hyaluronic acid, thiol hyaluronic acid, collagen ( collagen), lysine (Lysine), polyethyleneimine (PEI), glutathione (GSH) and the like can be used. At this time, the chitosan has a molecular weight of 1 ~ 220kDa, Pluronic weight average molecular weight of 8,700Da F68 or 12,600Da F127, hyaluronic acid is preferably used a weight average molecular weight of 230 ~ 2,000kDa.

In particular, as the bio / synthetic polymer having a —NH 2 or —SH group at the terminal, a compound represented by the following Chemical Formulas 5 to 7 may be used.

[Chemical Formula 5]

In Formula 5,

n + m is an integer of 10-700, n is an integer of 0-700.

[Chemical Formula 6]

In Formula 6,

x is an integer of 1-12, n is an integer of 78-100, m is an integer of 30-65.

[Formula 7]

In Formula 7,

x is an integer of 1-12, n is an integer of 78-100, m is an integer of 30-65.

In addition, the biocompatible polymer capable of crosslinking to the biocompatible polymer having a catechol-based functional group may be the same kind as described above.

The bio / synthetic polymer or biocompatible polymer is preferably included in the crosslinked hydrogel in an amount of 0.1 to 98% by weight, more preferably 1 to 50% by weight. If the content is included in the above range is better for crosslinking.

The biocompatible polymer having a catechol-based functional group is a biocompatible polymer having a catechol-based functional group in a TBS (pH 7.4) solvent in order to crosslink with a bio / synthetic polymer having a -NH 2 or -SH group or a biocompatible polymer. Dissolved in, dissolved in a TBS (pH 8.5) bio / synthetic polymer having a -NH2 or -SH group in a TBS (pH 8.5) solvent, and after mixing the two solutions to adjust the pH to more than 8.5, for a complete crosslinking reaction The mixed solution is maintained for 1 to 24 hours at a temperature of 0 ~ 80 ℃. Then, the crosslinked hydrogel may be centrifuged to remove impurities, prepared into a scaffold, and lyophilized to obtain a final crosslinked hydrogel.

Hereinafter, the present invention will be described in more detail with reference to examples. These embodiments are for purposes of illustration only and are not intended to limit the scope of protection of the present invention.

Example  1. Catechol system  Compounds having functional groups can be added to biocompatible polymers. By conjugation  Formed Bridge Hydrogel  Produce

(Catechol hyaluronic acid (HA-C) production)

1% by weight of hyaluronic acid having a molecular weight of about 230 kDa was completely dissolved in 2x PBS (phosphate buffered saline, pH 7.4) buffer. At this time, the buffer solution was used to remove oxygen and filled with nitrogen. 0.4% by weight of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) and 0.3% by weight of N-hydroxysuccinimide (NHS) were added to the solution, followed by pH using 1M HCl. Was adjusted to 3 to 4 degrees. Then, 0.4% by weight of dopamine (dopamine) was added and the pH was maintained at 3 to 4, and the mixed solution was stirred at room temperature for 24 hours. The mixed solution was sealed with a pre-washed dialysis membrane (molecular weight cut-off of 6-8 kDa) and dialyzed in a 2 × PBS solution of pH 3 or less for 48 hours to remove residual EDC, NHS and dopamine. Then, freeze-dried to give catechol hyaluronic acid (HA-C) (see Scheme 1 below).

[Reaction Scheme 1]

In Scheme 1, x + y is an integer of 200 to 6,000, y is an integer of 1 to 50.

(Manufacture of catechol hyaluronic acid crosslinked product)

The prepared catechol hyaluronic acid (HA-C) was completely dissolved in TBS (pH 8.5) buffer at a concentration of 60% by weight, and the mixed solution was maintained at room temperature for 24 hours for complete crosslinking reaction. Thereafter, the prepared catechol hyaluronic acid crosslinked product was centrifuged three times to remove impurities, and then prepared as a scaffold and lyophilized.

Example  2. Catechol hyaluronic acid ( HA -C) and chitosan derivatives ( Kitalac ) Bridge Hydrogel  Produce

(Chitosan derivative (chitalac) production)

2% by weight of chitosan with a molecular weight of about 5 kDa was completely dissolved in a 1% acetic acid (pH 4.5) solution, to which methanol was added in an amount equal to 1% acetic acid. Thereafter, 4.5 wt% of lactose and 2 wt% of sodium cyanoborohydride were dissolved in the same ratio of a mixed solvent of 1% acetic acid and methanol, and then added to the solution in which the chitosan was dissolved. The mixed solution was stirred at room temperature for 24 hours, sealed with a pre-washed dialysis membrane (molecular weight cut-off of 6-8 kDa), and dialyzed in secondary distilled water for 24 hours, followed by residual chitosan, lactose and sodium cyanoborohydride. Was removed. Then, lyophilization to obtain a chitosan derivative (chitalac) (see Scheme 2 below).

[Reaction Scheme 2]

In Scheme 2, n + m is an integer of 10 to 700, n is an integer of 1 to 700.

(Manufacture of catechol hyaluronic acid and chitosan derivative crosslinked hydrogel)

The catechol hyaluronic acid (HA-C) prepared in Example 1 was completely dissolved in TBS (pH 7.4) buffer solution at a concentration of 4 (w / w)%, and then prepared to crosslink by reaction with an amine. One chitosan derivative was dissolved in TBS (pH 8.5) at a concentration of 2 (w / w)%, and the pH was adjusted to 8.5 after mixing the two solutions. The mixed solution was maintained at room temperature for 24 hours for complete crosslinking of the catechol hyaluronic acid and chitosan derivative crosslinked hydrogel. Thereafter, the prepared catechol hyaluronic acid and chitosan derivative crosslinked hydrogels were subjected to centrifugation three times to remove impurities, and then made into scaffolds and lyophilized.

Experimental Example  1. Catechol Hyaluronic Acid and Chitosan Derivatives Bridge Hydrogel  Strength measurement

After centrifugation of the catechol hyaluronic acid and chitosan derivative crosslinked hydrogel prepared in Example 2 to remove the supernatant, the strength was confirmed by measuring the viscoelasticity of the hydrogel through a rheometer, and the results are shown in FIG. 1. It was. At this time, hyaluronic acid and chitosan derivative (chitalac) crosslinked hydrogel was used as a control.

As shown in Figure 1, the catechol hyaluronic acid and chitosan derivative crosslinked hydrogel prepared according to the present invention was confirmed that the strength is greater compared to the control hyaluronic acid and chitosan derivative crosslinked hydrogel.

Experimental Example  2. Catechol Hyaluronic Acid and Chitosan Derivatives Bridge Hydrogel  Intensity measurement over time

The time-dependent strength of the catechol hyaluronic acid and chitosan derivative crosslinked hydrogel prepared in Example 2 was measured through a Rheometer, and the results are shown in FIG. 2.

As shown in Figure 2, the catechol hyaluronic acid and chitosan derivative crosslinked hydrogel prepared according to the present invention was confirmed to change from the sol state to the gel state over time, without changing the strength over time It could be confirmed that maintaining a large strength.

Experimental Example  3. Catechol Hyaluronic Acid and Chitosan Derivatives Bridge Hydrogel  Swelling

The catechol hyaluronic acid and chitosan derivative crosslinked hydrogels prepared in Example 2 were immersed in distilled water and stored in a 37 ° C. incubator, and then the weight of the crosslinked hydrogels was wiped with a dust-free tissue. The weight of the catechol hyaluronic acid and chitosan derivative crosslinked hydrogel was measured before the immersion. At this time, the degree of swelling of the crosslinked hydrogel was calculated by the following Equation 1, and the percent swelling through the swelling degree was calculated by the following Equation 2.

[Equation 1]

&Quot; (2) "

As shown in FIG. 3, the catechol hyaluronic acid (HA-C) and chitosan derivative (chitalac) crosslinked hydrogel prepared according to the present invention showed that the hydrogel absorbed water through the swelling degree. .

Experimental Example  4. Catechol Hyaluronic Acid and Chitosan Derivatives Bridge Hydrogel  Gel strength measurement under swelling

Swelling over time was measured using the catechol hyaluronic acid and chitosan derivative crosslinked hydrogel prepared in Example 2, and the gel strength at that time was measured through a rheometer, and the results are shown in FIG. 4.

Experimental results, as shown in Figure 4, the catechol hyaluronic acid and chitosan derivative crosslinked hydrogel prepared according to the present invention was confirmed to maintain a similar gel strength even after 20 days.

Example  3. Catechol hyaluronic acid ( HA -C) and amines Pluronic (F127- EDA ) Bridge Hydrogel  Produce

(Amine Pluronic (F127-EDA))

An amine pluronic (F127-EDA) was synthesized in which both ends of the pluronic were substituted with amines. After dissolving 11% by weight of Pluronic, 0.5% by weight of 4-dimethylaminopyridine (DMAP) and 0.5% by weight of triethylamine (TEA) in 1,4-dioxene, the succinic and hydride (SA) ) 0.5% by weight was added. The mixed solution was stirred at room temperature for 24 hours, the solvent was removed, and then precipitated in cold diethyl ether. The resulting precipitate was filtered and dried to give a white precipitate. The precipitate thus obtained was dissolved in N, N-dimethylformamide (DMF), followed by 0.4% by weight of N, N'-dicyclohexylcarbodiimide (DCC) and 0.2% by weight of N-hydroxysuccinimide (NHS). Added. Thereafter, 0.2 wt% of ethylenediamine and N, N-dimethylformamide (DMF) were slowly added to the mixed solution as a solvent. The mixed solution was stirred at 30 ° C. for 24 hours, the solvent was removed, and then precipitated in cold diethyl ether. The precipitate thus produced was filtered and dried to give a white precipitate. This precipitate was an amine pluronic (F127-EDA) with a weight average molecular weight of about 12,600 Da (see Scheme 3 below).

Scheme 3

In Scheme 3, x is an integer of 1 to 12, n is an integer of 78 to 100, m is an integer of 30 to 65.

(Catechol Hyaluronic Acid and Amine Pluronic Crosslinked Hydrogel Preparation)

After dissolving catechol hyaluronic acid (HA-C) having a molecular weight of about 230 kDa prepared in Example 1 at a concentration of 5 (w / w)% in TBS (pH 7.4) buffer solution, crosslinking was performed by reaction with amine. To achieve this, the prepared amine pluronic (F127-EDA) was dissolved in TBS (pH 8.5) at various concentrations ranging from 15 to 25 (w / w)%, and the two solutions were mixed and adjusted to pH 8.5. The mixed solution was maintained at 4 ° C. for 24 hours for complete crosslinking of the catechol hyaluronic acid with the amine pluronic crosslinked hydrogel. Thereafter, the prepared catechol hyaluronic acid and the amine pluronic crosslinked hydrogel were subjected to centrifugation three times to remove impurities, and then made into a scaffold and lyophilized.

Experimental Example  5. Catechol Hyaluronic Acid and Amine Pluronic Bridge Hydrogel  Strength measurement

Using the catechol hyaluronic acid and amine pluronic crosslinked hydrogel prepared in Example 3, the strength of the hydrogel was measured through a rheometer over time, and the results are shown in FIG. 5.

As a result, as shown in FIG. 5, the catechol hyaluronic acid and amine pluronic crosslinked hydrogel prepared according to the present invention was sol at the intersection of G 'and G' '.

Phase change to gel was confirmed, and catechol hyaluronic acid and amine pluronic crosslinked hydrogel showed great strength.

Example  4. Hyaluronic acid ( HA ) And catechol amines ( EDA -C) Bridge Hydrogel

(Manufacture of catechol amine (EDA-C))

A catechol group was synthesized at both ends of the ethylene diamine. After dissolving 0.5% by weight of ethylenediamine in N, N-dimethylformamide (DMF), 0.2% by weight of N, N'-dicyclohexylcarbodiimide (DCC) and N-hydroxysuccinimide (NHS) 0.2 Add weight percent. Subsequently, 0.2 wt% of hydrocaffeic acid was added to the mixed solution. The mixed solution was stirred at room temperature for 24 hours, the solvent was removed, and then precipitated in cold diethyl ether. The resulting precipitate was filtered and dried to give a white precipitate. This precipitate was catechol amine (EDA-C).

(Preparation of hyaluronic acid and catechol amine crosslinked hydrogel)

Hyaluronic acid having a molecular weight of about 230 kDa was completely dissolved in TBS (pH 7.4) buffer at a concentration of 5 (w / w)%, and the catechol amine (EDA-C) prepared above was cross-linked by reaction with an amine. It was dissolved in TBS (pH 8.5) at various concentrations ranging from 15-25 (w / w)%, and the pH was adjusted to 8.5 after mixing the two solutions. The mixed solution was maintained at 4 ° C. for 24 hours for complete crosslinking of hyaluronic acid (HA) and catechol amine (EDA-C) crosslinked hydrogels. Then, the hyaluronic acid (HA) and catechol amine (EDA-C) cross-linked hydrogel was centrifuged three times to remove impurities, and then made into a scaffold and lyophilized.

Example   5.Jekcatechol hyaluronic acid ( HA -C) and Thiol Pluronic (F127- CA ) Bridge Hydrogel  Produce

(Manufactured by Thiol Pluronic (F127-CA))

Thiol Pluronic (F127-CA) was synthesized in which both ends of the pluronic were substituted with thiols. Molecular weight of about 12 kDa Pluronic 11%, 4-dimethylaminopyridine (DMAP) 0.5% by weight and triethylamine (TEA) 0.5% by complete dissolution in 1,4-dioxene, then succinic and hydride (SA) 0.5 wt% was added. The mixed solution was stirred at room temperature for 24 hours, the solvent was removed, and then precipitated in cold diethyl ether. The resulting precipitate was filtered and dried to give a white precipitate. The precipitate thus obtained was dissolved in N, N-dimethylformamide (DMF), followed by 0.4% by weight of N, N'-dicyclohexylcarbodiimide (DCC) and 0.2% by weight of N-hydroxysuccinimide (NHS). Added. Thereafter, 0.2 wt% of cystiamine was added to the mixed solution. The mixed solution was stirred at 30 ° C. for 24 hours, the solvent was removed, and then precipitated in cold diethyl ether. The precipitate thus produced was filtered and dried to give a white precipitate. This precipitate was thiol pluronic (F127-CA) with a weight average molecular weight of 12,600 Da (see Scheme 4 below).

[Reaction Scheme 4]

In Scheme 4, n is an integer of 78 to 100, m is an integer of 30 to 65.

Preparation of Catechol Hyaluronic Acid and Thiol Pluronic Crosslinked Hydrogel

After dissolving catechol hyaluronic acid (HA-C) having a molecular weight of about 230 kDa prepared in Example 1 at a concentration of 5 (w / w)% in TBS (pH 7.4) buffer solution, it was crosslinked by reaction with thiol. To achieve the above, the prepared thiol pluronic (F127-CA) was dissolved in TBS (pH 8.5) at various concentrations ranging from 15 to 25 (w / w)%, and the pH of the solution was adjusted to 8.5 after mixing the two solutions. Adjusted. The mixed solution was maintained at 4 ° C. for 24 hours for complete crosslinking of the catechol hyaluronic acid with the thiol pluronic crosslinked hydrogel. Thereafter, the prepared catechol hyaluronic acid and the thiol pluronic crosslinked hydrogel were subjected to centrifugation three times to remove impurities, and then made into a scaffold and lyophilized.

Experimental Example  6. Catechol Hyaluronic Acid Thiol Pluronic Bridge Hydrogel  Strength measurement

Using the catechol hyaluronic acid and thiol pluronic crosslinked hydrogel prepared in Example 6, the strength of the hydrogel was measured through a rheometer over time, and the results are shown in FIG. 6.

As a result, as shown in Figure 6, the catechol hyaluronic acid and the thiol pluronic crosslinked hydrogel prepared in accordance with the present invention was applied to sol at the intersection of G 'and G' '.

It was confirmed that the phase change to the gel occurs, these showed a large intensity.

Example  6. Catechol Chitlac-C  Amine Pluronic (F127- EDA ) Bridge Hydrogel  Produce

(Made by catechol chitlac-C)

0.5 wt% of the chitosan derivative (chitalac) prepared in Example 2 was completely dissolved in secondary distilled water (pH 5.5). After 0.2% by weight of hydrocaffeic acid and 0.2% by weight of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) were completely dissolved in a mixture of secondary distilled water and ethanol, the chitosan derivative was used. Was added to the dissolved mixed solution. The mixed solution was stirred at room temperature for 12 hours while maintaining pH 5.5 with 1M HCl. Then, it was sealed with a pre-washed dialysis membrane (molecular weight cut-off of 3 kDa) and dialyzed with secondary distilled water (pH 5.5) for 2 days, followed by residual hydrocaffeic acid and 1-ethyl-3- (3-dimethylaminopropyl). Carbodiimide (EDC) was removed. It was then lyophilized to give catechol chitalac (Chitlac-C) (see Scheme 5 below).

[Reaction Scheme 5]

In Scheme 5, Z is a compound having a catechol-based functional group, n + m is an integer of 10 to 700, n is an integer of 1 to 699, x is an integer of 1 to 700.

(Catechol chitalac and amine pluronic crosslinked hydrogel preparation)

The catechol chitalac (Ctlac-C) having a molecular weight of about 12 kDa was completely dissolved in TBS (pH 7.4) buffer at a concentration of 0.5 (w / w)%, and then carried out to crosslink by reaction with an amine. Amine Pluronic (F127-EDA) prepared in Example 3 was dissolved in TBS (pH 8.5) at various concentrations ranging from 15-25 (w / w)%, and the pH was adjusted to 8.5 after mixing the two solutions. . The mixed solution was maintained at 4 ° C. for 24 hours for complete crosslinking of the catechol chitalac and amine pluronic crosslinked hydrogel. Next, the catechol chitalac and amine pluronic crosslinked hydrogels were subjected to centrifugation three times to remove impurities, and then lyophilized by preparing a scaffold.

Experimental Example  7. Catechol Kitalac  Amine Pluronic Bridge Hydrogel  Strength measurement

Using the catechol chitalac and amine pluronic crosslinked hydrogel prepared in Example 6, the strength of the hydrogel was measured through a Rheometer over time, and the results are shown in FIG. 7.

As a result, as shown in Figure 7, the catechol chitalac and amine pluronic crosslinked hydrogel prepared according to the present invention was confirmed that the phase change from sol to gel at the intersection of G 'and G' '. It was confirmed that these showed great strength.

Example  7. Catechol Chitlac-C  Amine hyaluronic acid ( HA - EDA ) Bridge Hydrogel  Produce

(Amine hyaluronic acid (HA-EDA) production)

0.5% by weight of hyaluronic acid having a molecular weight of about 230 kDa was completely dissolved in 2x PBS (phosphate buffered saline, pH 7.4) buffer. At this time, the buffer solution was removed to oxygen and filled with nitrogen. 0.2% by weight of ethylenediamine was added after adding 0.2% by weight of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) and 0.2% by weight of N-hydroxysuccinimide (NHS) to the solution. Was added and the mixed solution was stirred at room temperature for 24 hours. The mixed solution was sealed with a pre-washed dialysis membrane (molecular weight cut-off of 6-8 kDa) and dialyzed for 48 hours in a 2x PBS solution to remove residual EDC, NHS and ethylenediamine. Lonic acid (HA-EDA) was obtained.

(Manufacture of catechol chitalac and amine hyaluronic acid crosslinked hydrogel)

The catechol chitalac having a molecular weight of about 12 kDa prepared in Example 6 was completely dissolved in TBS (pH 7.4) buffer at a concentration of 0.5 (w / w)%, and then 15-25 (w / w) amine hyaluronic acid. It was dissolved in TBS (pH 8.5) at various concentrations in the range of%, and the pH was adjusted to 8.5 after mixing the two solutions. The mixed solution was maintained at room temperature for 24 hours for complete crosslinking reaction between catechol chitalac and amine hyaluronic acid crosslinked hydrogel. Thereafter, the catechol chitalac and amine hyaluronic acid crosslinked hydrogels were subjected to centrifugation three times to remove impurities, and then made into scaffolds and lyophilized.

Example  8. Catechol Chitlac-C Thiol Pluronic (F127- CA ) Bridge Hydrogel  Produce

The catechol chitalac having a molecular weight of about 12 kDa prepared in Example 6 was completely dissolved in TBS (pH 7.4) buffer solution at a concentration of 0.5 (w / w)%, and then subjected to crosslinking by reaction with thiol. The thiol pluronics prepared in Example 5 were dissolved in TBS (pH 8.5) at various concentrations ranging from 15-25 (w / w)%, and the pH was adjusted to 8.5 after mixing the two solutions. The mixed solution was maintained at 4 ° C. for 24 hours for complete crosslinking of the catechol chitalac and thiol pluronic crosslinked hydrogel. Then, the catechol chitalac and thiol pluronic crosslinked hydrogel were subjected to centrifugation three times to remove impurities, and then made into a scaffold and lyophilized.

Experimental Example  8. Catechol Kitalac Thiol Pluronic Bridge Hydrogel  Strength measurement

Using the catechol chitalac and thiol pluronic crosslinked hydrogel prepared in Example 8, the strength of the hydrogel was measured through a Rheometer with time, and the results are shown in FIG. 8.

As a result of the experiment, as shown in Figure 8, the catechol chitalac and thiol pluronic crosslinked hydrogel prepared according to the present invention can be seen that the phase change from sol to gel at the intersection of G 'and G''. It was confirmed that they showed a great strength.

Example  9. Catechol chitosan ( chitosan -C) and amines Pluronic (F127- EDA ) Bridge Hydrogel  Produce

(Catechol chitosan (C) production)

0.5% by weight of chitosan was completely dissolved in secondary distilled water (pH 5.5). After dissolving 0.2% by weight of hydrocaffeic acid and 0.2% by weight of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) in a mixture of secondary distilled water and ethanol in the same ratio, the chitosan was dissolved. To the prepared mixed solution. The mixed solution was stirred at room temperature for 12 hours while maintaining pH 5.5 with 1M HCl. Then, it was sealed with a pre-washed dialysis membrane (molecular weight cut-off of 3 kDa) and dialyzed with secondary distilled water (pH 5.5) for 2 days, followed by residual hydrocaffeic acid and 1-ethyl-3- (3-dimethylaminopropyl). Carbodiimide (EDC) was removed. Then, lyophilization to give catechol chitosan (chitosan-C) (see Scheme 6 below).

[Reaction Scheme 6]

In Scheme 6, Z is a compound having a catechol-based functional group, n is an integer of 10 to 700, m is an integer of 1 to 699.

(Catechol Chitosan and Amine Pluronic Crosslinked Hydrogel Preparation)

The amine prepared in Example 3 was dissolved in a catechol chitosan having a molecular weight of about 9 kDa in a TBS (pH 7.4) buffer solution at a concentration of 0.5 (w / w)% and then crosslinked by reaction with an amine. Pluronic was dissolved in TBS (pH 8.5) at various concentrations ranging from 15-25 (w / w)%, and the pH was adjusted to 8.5 after mixing the two solutions. The mixed solution was maintained at 4 ° C. for 24 hours for complete crosslinking of the catechol chitosan and the amine pluronic crosslinked hydrogel. Thereafter, the prepared catechol chitosan and the amine pluronic crosslinked hydrogel were subjected to centrifugation three times to remove impurities, and then made into a scaffold and lyophilized.

Example  10. Catechol chitosan ( chitosan -C) and amine hyaluronic acid ( HA - EDA ) Bridge Hydrogel  Produce

After dissolving the catechol chitosan having a molecular weight of about 12 kDa prepared in Example 9 in a concentration of 0.5 (w / w)% in TBS (pH 7.4) buffer solution, the amine hyaluronic acid was 15-25 (w / w)% It was dissolved in TBS (pH 8.5) at various concentrations in the range, and the pH was adjusted to 8.5 after mixing the two solutions. The mixed solution was maintained at room temperature for 24 hours for complete crosslinking of the catechol chitosan and the amine hyaluronic acid crosslinked hydrogel. Thereafter, the prepared catechol chitosan and amine hyaluronic acid crosslinked hydrogels were subjected to centrifugation three times to remove impurities, and then made into a scaffold and lyophilized.

Example  11.Catechol chitosan ( chitosan -C) and Thiol Pluronic  (F127- CA ) Bridge Hydrogel  Produce

The catechol chitosan having a molecular weight of about 9 kDa prepared in Example 9 was completely dissolved in TBS (pH 7.4) buffer solution at a concentration of 0.5 (w / w)%, and then cross-linked by reaction with thiol. The thiol pluronic prepared in 5 was dissolved in TBS (pH 8.5) at various concentrations ranging from 15 to 25 (w / w)%, and the pH was adjusted to 8.5 after mixing the two solutions. The mixed solution was maintained at 4 ° C. for 24 hours for complete crosslinking of the catechol chitosan and the thiol pluronic crosslinked hydrogel. Then, catechol chitosan and thiol pluronic crosslinked hydrogel were centrifuged three times to remove impurities, and then scaffolded and lyophilized.

Example  12. Catechol With amines (EDA-C)  Chitosan derivatives ( chitlac ) Bridge Hydrogel  Produce

The chitosan derivative (chitalac) prepared in Example 2 was completely dissolved in TBS (pH 8.5) buffer solution at a concentration of 5 (w / w)%, and then in Example 4 to form a crosslink by reaction with an amine. The prepared catechol amine was dissolved in TBS (pH 7.4) buffer at various concentrations ranging from 15-25 (w / w)%, and the two solutions were mixed and adjusted to pH 8.5. The mixed solution was maintained at room temperature for 24 hours for complete crosslinking of the catechol amine and the chitosan derivative crosslinked hydrogel. The catechol amine and chitosan derivative crosslinked hydrogels were then centrifuged three times to remove impurities, and then scaffolded and lyophilized.

Example  13. Catechol With amines (EDA-C)  Chitosan ( chitosan ) Bridge Hydrogel  Produce

Chitosan was dissolved in TBS (pH 8.5) buffer completely at a concentration of 5 (w / w)%, and the catechol amine prepared in Example 4 was crosslinked by reaction with amine 15-25. It was dissolved in TBS (pH 7.4) buffer at various concentrations in the range of (w / w)%, and the pH was adjusted to 8.5 after mixing the two solutions. The mixed solution was maintained at room temperature for 24 hours for complete crosslinking of the catechol amine and the chitosan crosslinked hydrogel. Then, the catechol amine and chitosan crosslinked hydrogel were subjected to centrifugation three times to remove impurities, and then lyophilized by preparing a scaffold.

Experimental Example  9. Bridge Hydrogel  Strength measurement

The strength of each of the crosslinked product and the crosslinked product hydrogel prepared in Examples 2 to 14 was measured, and the results are shown in FIG. 9.

As shown in FIG. 9, the G 'value of each hydrogel was about 600 or more, and it was found that the crosslinked hydrogels prepared in Examples 2 to 13 had strong strengths. .

Claims (15)

In the crosslinked hydrogel comprising a compound having a catechol-based functional group as an active ingredient,
Iii) formed by crosslinking of compounds having catechol-based functional groups,
Ii) a compound having a catechol-based functional group is conjugated to a biocompatible polymer, or
Iii) formed by crosslinking of a bio / synthetic polymer having a catechol-based functional group and a bio / synthetic polymer having a -NH 2 or -SH group at its terminal;
Iii) a crosslinked hydrogel formed by crosslinking a bio / synthetic polymer having a catechol-based functional group and a -NH2 or -SH group, and a bio / synthetic polymer having a -NH2 or -SH group at its terminal.
The method of claim 1,
The compound having a catechol-based functional group is 2-chloro-3 ', 4'-dihydroxyacetophenone, dopa, dopamine, caffeic acid, garlic acid, chlorogenic acid, (-)-epicatechin, (-)-epicatechin Gallate, melanin, tannin, flavonoids, cyanidin, dihydro quercetin, oron, dihydromyricetin, delfinidine, mycetin, quercetin, (-)-epicalatechin-3-gallate, (-)-epigallo Crosslinked hydrogel, characterized in that any one or more selected from catechin, (-)-epigallocatechin gallate, catechin and luteolin.
The method of claim 1,
The biocompatible polymer,
Hyaluronic acid, chitosan, chitosan derivatives (chitalac), collagen, collagen, fibronectin, collagen, gelatin, alginic acid, pluronic , poly (lacticacid) (PLA), poly (glycolicacid) (PGA), poly (D, L-latide-co-glycolide) (PLGA), poly (ε-caprolactone) (PCL), polyanhydride, poly Crosslinked hydrogel, characterized in that at least any one selected from esters (polyester), lysine (Lysine), Polyethyleneimine (PEI) and glutathione (GSH).
The method of claim 1,
Bio-synthetic polymer having a catechol-based functional group is a crosslinked hydrogel, characterized in that any one or more of the compounds represented by the following formula (1) to:
[Chemical Formula 1]

In Formula 1,
x + y is an integer from 200 to 6,000, y is an integer from 1 to 50,
(2)

In Formula 2,
A is SH, NH2 functional group, n is 1-3
x + y is an integer from 200 to 6,000, y and z are integers from 1 to 50
(3)

In Formula 3,
Z is a compound having a catechol-based functional group,
n + m is an integer from 10 to 700,
n is an integer from 0 to 699, m is an integer from 1 to 700, x is from 1 to 700
Is an integer of
[Chemical Formula 4]

In Formula 4,
Z is a compound having a catechol-based functional group,
n + m is an integer from 10 to 700,
n and m are each an integer of 1-700.
The method of claim 1,
Bio-synthetic polymer having the -NH2 or -SH group is chitosan, chitosan derivative (chitalac), amine pluronic, catechol amine, thiol pluronic, amine hyaluronic acid, thiol hyaluronic acid, lysine (Lysine) Crosslinked hydrogel, characterized in that at least one selected from Polyethyleneimine (PEI) and glutathione (GSH).
The method of claim 1,
The bio-synthetic polymer having the -NH2 or -SH group is a crosslinked hydrogel, characterized in that any one or more of the compounds represented by the following formulas (6) to (8):
[Chemical Formula 5]

In Formula 5,
n + m is an integer from 10 to 700, n is an integer from 1 to 700,
[Chemical Formula 6]

In Formula 6,
x is an integer from 1 to 12, n is an integer from 78 to 100, m is from 30 to
Is an integer of 65,
(7)

In Formula 7,
x is an integer from 1 to 12, n is an integer from 78 to 100, m is 30 to 65
Is an integer.
The method of claim 1,
The bio / synthetic polymer having the catechol-based functional group is formed by conjugating the compound having the catechol-based functional group with the bio / synthetic polymer.
8. The method of claim 7,
Iii) in the crosslinked hydrogel of
The compound having a catechol-based functional group is contained in 0.1 to 99.9% by weight, the bio-synthetic polymer having a NH2 or SH functional group is a crosslinked hydrogel, characterized in that contained in 0.1 to 99.9% by weight.
The method of claim 1,
The bio-synthetic polymer having the catechol-based functional group and the NH2 or -SH group is formed by conjugating the compound having the catechol-based functional group with the bio / synthetic polymer having the NH2 or -SH group.
The method of claim 9,
Iii) in the crosslinked hydrogel of
The catechol-based functional group and the compound having NH 2 or SH functional group is included in 0.1 to 98% by weight, the bio / synthetic polymer having the NH 2 or -SH group is included in 0.1 to 98% by weight, and -NH 2 or-at the terminal Bio-synthetic polymer having a SH group is a crosslinked hydrogel, characterized in that contained in 1 to 50% by weight.
The method of claim 1,
The crosslinked hydrogel is crosslinked hydrogel, characterized in that formed at 20 to 80 ℃.
In the method for producing a crosslinked hydrogel comprising a compound having a catechol-based functional group as an active ingredient,
Mixing a solution in which a bio / synthetic polymer having a catechol-based functional group is dissolved in a solvent and a solution in which a bio / synthetic polymer having a -NH 2 or -SH group is dissolved in a solvent;
maintaining at a pH of at least 8.5; And
Method for producing a crosslinked hydrogel, characterized in that it comprises a step 3 to remove impurities by centrifugation.
13. The method of claim 12,
Bio-synthetic polymer having a catechol-based functional group,
To the solution in which the bio / synthetic polymer is dissolved in a solvent, the compound having the catechol-based functional group is added and reacted in a nitrogen atmosphere under acidic conditions of pH 5 or less, and then dialyzed under acidic conditions of pH 4 or less to form bio / synthetic polymer. Method for producing a crosslinked hydrogel, characterized in that the catechol group is formed by conjugation.
13. The method of claim 12,
Step 2,
Method for producing a crosslinked hydrogel, characterized in that maintained for 1 to 24 hours at a temperature of 20 to 80 ℃.
13. The method of claim 12,
Step 3,
Method of producing a cross-linked hydrogel, characterized in that it further comprises the step of producing a freeze-dried scaffold.

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