JP6556208B2 - Hydrogel composition and drug delivery system comprising the same - Google Patents

Description

CROSS REFERENCE TO RELATED APPLICATIONS This application is a benefit of US Provisional Patent Application No. 62 / 410,165 filed on October 21, 2016 and priority of Taiwan Patent Application No. 106135658 filed on October 18, 2017. And are hereby incorporated by reference in their entirety.

TECHNICAL FIELD The technical field relates to novel hydrogel compositions and drug delivery systems comprising the same.

  Currently popular drug delivery carriers cannot carry high concentrations of drug. However, the use of many antibody pharmaceuticals requires relatively high doses to be effective. Therefore, the controlled release technology of antibody drugs faces difficult problems. The production of a general carrier requires the use of an organic solvent.

U.S. Pat. No. 8519086 Taiwan Patent Application No. 201310131 Specification Taiwan Patent Application No. 2006262626 Specification Chinese Patent No. 10684546A Specification

  Organic solvents often lead to protein inactivation. Furthermore, it is difficult to maintain the molecular structure of the drug and the integrity and stability of the biological activity during the drug release period.

  Therefore, there is a need for the development of carriers that can carry high concentrations of drug and that can maintain the molecular structure of the loaded drug and the integrity and stability of the biological activity during the drug release period.

  According to one embodiment of the present disclosure, a hydrogel composition is provided. The hydrogel composition includes polyglutamic acid (PGA) containing maleimide groups and polyethylene glycol (PEG) containing terminal thiol groups. In particular, the pH value of the hydrogel composition is about 4.0 to about 6.5.

  According to another embodiment of the present disclosure, a drug delivery system is provided. The drug delivery system includes the hydrogel composition and a pharmaceutically active ingredient encapsulated within the hydrogel composition.

More specifically, the present invention provides the following:
[1]
A hydrogel composition comprising:
Polyglutamic acid (PGA) containing maleimide groups;
A hydrogel composition comprising polyethylene glycol (PEG) containing a terminal thiol group, wherein the pH value of the hydrogel composition is 4.0 to 6.5;
[2]
The hydrogel composition according to [1], wherein the polyglutamic acid (PGA) containing the maleimide group has a molecular weight of 10 kDa to 1000 kDa.
[3]
The hydrogel composition according to one of [1] and [2], wherein a graft ratio of the polyglutamic acid (PGA) containing the maleimide group is 5% to 40%;
[4]
The concentration of polyglutamic acid (PGA) containing the maleimide group in the hydrogel composition is 0.75 wt% to 10 wt%, according to one of [1], [2] and [3] A hydrogel composition;
[5]
The hydrogel composition according to one of [1], [2], [3] and [4], wherein the polyglutamic acid (PGA) containing a maleimide group does not contain a thiol group;
[6]
The hydrogel composition according to one of [1], [2], [3], [4] and [5], wherein the polyethylene glycol (PEG) containing the terminal thiol group has a molecular weight of 2 kDa to 20 kDa. object;
[7]
The concentration of polyethylene glycol (PEG) containing the terminal thiol group in the hydrogel composition is 0.75 wt% to 10 wt% [1], [2], [3], [4], [5 ] And a hydrogel composition according to one of [6];
[8]
[1], [2], [3], [4], [5], [6] and the polyethylene glycol (PEG) containing the terminal thiol group is a 4-arm type, an 8-arm type or a Y-form [7] The hydrogel composition according to one of the above;
[9]
[1], [2], [3], [4], [5], [6], [7] and [8], wherein the polyethylene glycol (PEG) containing the terminal thiol group does not contain a maleimide group A hydrogel composition according to one of the preceding;
[10]
The molar ratio of the thiol group to the maleimide group in the polyethylene glycol (PEG) containing the terminal thiol group and the polyglutamic acid (PGA) containing the maleimide group is 0.2 to 5.0, [1] , [2], [3], [4], [5], [6], [7], [8] and [9].
[11]
The molar ratio of the thiol group to the maleimide group in the polyethylene glycol (PEG) containing the terminal thiol group and the polyglutamic acid (PGA) containing the maleimide group is 1.0 to 1.5, [1] , [2], [3], [4], [5], [6], [7], [8], [9] and [10] according to one of the hydrogel compositions;
[12]
Described in one of [1], [2], [3], [4], [5], [6], [7], [8], [9], [10] and [11] A hydrogel composition of
A drug delivery system comprising a pharmaceutically active ingredient encapsulated in the hydrogel composition;
[13]
The drug delivery system according to [12], wherein the pharmaceutically active ingredient is selected from the group consisting of growth factors, hormones, peptides, proteins, antibodies, hydrophilic small molecules and hydrophobic small molecules;
[14]
The drug delivery system according to one of [12] and [13], wherein the pharmaceutically active ingredient is selected from the group consisting of intact antibodies and antibody fragments;
[15]
The drug according to one of [12], [13] and [14], wherein the pharmaceutically active ingredient is selected from the group consisting of a mouse antibody, a chimeric antibody, a humanized antibody, and a human antibody Delivery system;
[16]
The drug delivery system according to one of [12] and [13], wherein the pharmaceutically active ingredient is selected from the group consisting of an antitumor drug, an antipsychotic drug, an analgesic and an antibiotic;
[17]
[12], [13], [14], [15] and [16], wherein the pharmaceutically active ingredient further associates with a polymer, metal, charged compound or charged particle to form a complex thereof One drug delivery system according to one;
[18]
The drug delivery system according to [17], wherein the polymer includes polyglutamic acid (PGA), hyaluronic acid, chitosan, or dextran;
[19]
The drug delivery system according to [17], wherein the metal includes zinc, calcium, magnesium, or iron;
[20]
The drug delivery system of [17], wherein the size of the complex is between 10 nm and 100 μm; and
[21]
The drug delivery system according to one of [12], [17] and [20], wherein the concentration of the pharmaceutically active ingredient or the complex is 1 mg / mL to 300 mg / mL.

  Poly (γ-glutamic acid) (γ-PGA) is a high molecular weight polypeptide composed of a γ-bonded glutamic acid unit and an α-carboxylate side chain. γ-PGA has many advantages such as non-toxicity, hydrophilicity, biodegradability, and avoidance of antigenicity or immunogenicity, making it an ideal biomaterial applicable to make functional hydrogel systems It is. In addition, abundant carboxyl groups on the γ-PGA chain can be chemically modified and can bind to soluble antibody molecules by electrostatic interactions.

  Poly (ethylene glycol) (PEG) as an uncharged hydrophilic segment is widely used for building chemically or physically cross-linked hydrogel systems because of its biocompatibility. PEG-based hydrogels have been utilized as cell scaffolds, adhesives for medical applications, and delivery vehicles. In particular, the ability to adjust the crosslink density provides flexibility and adaptability to PEG-based hydrogels and can be used for cell encapsulation and tissue growth.

  In this disclosure, in order to achieve high drug loading (greater than 150 mg / mL) and long-term sustained release (greater than 21 days), it is subcutaneously injectable and indium composed of γ-PGA and 4-arm PEG. Develop a situ-formed hydrogel system. γ-PGA is partially modified with N- (2-aminoethyl) maleimide by aminolization. The resulting maleimide-containing γ-PGA (γ-PGA-MA) and 4-arm PEG-SH are mixed in an aqueous solution. A chemically crosslinked hydrogel is formed through a pH-sensitive Michael addition reaction between maleimide and thiol groups.

  The following embodiments will be described in detail with reference to the accompanying drawings.

The present disclosure can be more fully understood by reading the following detailed description and examples with reference to the accompanying drawings.
FIG. 5 is a diagram illustrating a serum concentration-time profile according to an embodiment of the present disclosure. FIG. 3 is a diagram illustrating a tumor volume change curve according to an embodiment of the present disclosure.

DETAILED DESCRIPTION In the following detailed description, for the purposes of explanation, numerous specific details are set forth in order to provide a thorough understanding of the disclosed embodiments. However, it will be apparent that one or more embodiments may be practiced without these specific details. In other instances, well-known structures and devices are schematically shown in order to simplify the drawing.

  According to one embodiment of the present disclosure, a hydrogel composition is provided. The hydrogel composition includes polyglutamic acid (PGA) containing maleimide groups and polyethylene glycol (PEG) containing terminal thiol groups. In particular, the pH value of the hydrogel composition is about 4.0 to about 6.5.

In some embodiments, the polyglutamic acid (PGA) containing maleimide groups is, for example, as shown below and has a molecular weight of about 10 kDa to about 1000 kDa.

  In some embodiments, the graft rate of polyglutamic acid (PGA) containing maleimide groups is from about 5% to about 40%.

  In some embodiments, the concentration of polyglutamic acid (PGA) containing maleimide groups in the hydrogel composition is about 0.75 wt% to about 10 wt%.

  In some embodiments, polyglutamic acid (PGA) containing maleimide groups does not contain thiol groups.

  In some embodiments, the molecular weight of polyethylene glycol (PEG) containing terminal thiol groups is from about 2 kDa to about 20 kDa.

  In some embodiments, the concentration of polyethylene glycol (PEG) containing terminal thiol groups in the hydrogel composition is from about 0.75 wt% to about 10 wt%.

In some embodiments, the polyethylene glycol (PEG) containing a terminal thiol group can be, for example, a 4-arm type, an 8-arm type, or a Y form as shown below.

  In some embodiments, the polyethylene glycol (PEG) containing a terminal thiol group does not contain a maleimide group.

  In some embodiments, the molar ratio of thiol groups to maleimide groups of polyethylene glycol (PEG) containing terminal thiol groups and polyglutamic acid (PGA) containing maleimide groups is from about 0.2 to about 5.0. It is.

  In some embodiments, the molar ratio of thiol groups to maleimide groups of polyethylene glycol (PEG) containing terminal thiol groups and polyglutamic acid (PGA) containing maleimide groups is from about 1.0 to about 1.5. It is.

  According to another embodiment of the present disclosure, a drug delivery system is provided. The drug delivery system includes a hydrogel composition and a pharmaceutically active ingredient encapsulated within the hydrogel composition.

  In some embodiments, the hydrogel composition may include polyglutamic acid (PGA) containing maleimide groups and polyethylene glycol (PEG) containing terminal thiol groups. The pH value of the hydrogel composition is about 4.0 to about 6.5.

  In some embodiments, the molecular weight of polyglutamic acid (PGA) containing maleimide groups is from about 10 kDa to about 1000 kDa.

  In some embodiments, the graft rate of polyglutamic acid (PGA) containing maleimide groups is from about 5% to about 40%.

  In some embodiments, the concentration of polyglutamic acid (PGA) containing maleimide groups in the hydrogel composition is about 0.75 wt% to about 10 wt%.

  In some embodiments, polyglutamic acid (PGA) containing maleimide groups does not contain thiol groups.

  In some embodiments, the molecular weight of polyethylene glycol (PEG) containing terminal thiol groups is from about 2 kDa to about 20 kDa.

  In some embodiments, the concentration of polyethylene glycol (PEG) containing terminal thiol groups in the hydrogel composition is from about 0.75 wt% to about 10 wt%.

  In some embodiments, the polyethylene glycol (PEG) containing a terminal thiol group can be a 4-arm type, an 8-arm type, or a Y form.

  In some embodiments, the polyethylene glycol (PEG) containing a terminal thiol group does not contain a maleimide group.

  In some embodiments, the molar ratio of thiol groups to maleimide groups of polyethylene glycol (PEG) containing terminal thiol groups and polyglutamic acid (PGA) containing maleimide groups is about 0.2 to about 5.0. is there.

  In some embodiments, the molar ratio of thiol groups to maleimide groups of polyethylene glycol (PEG) containing terminal thiol groups and polyglutamic acid (PGA) containing maleimide groups is from about 1.0 to about 1.5. is there.

  In some embodiments, the pharmaceutically active ingredient can be selected from the group consisting of peptides, proteins, growth factors, hormones, antibodies, and hydrophilic or hydrophobic small molecules.

  In some embodiments, the pharmaceutically active ingredient can be selected from the group consisting of intact antibodies and antibody fragments.

  In some embodiments, the pharmaceutically active ingredient can be selected from the group consisting of a murine antibody, a chimeric antibody, a humanized antibody, and a human antibody.

  In some embodiments, the pharmaceutically active ingredient may be selected from the group consisting of an anti-tumor drug, an antipsychotic drug, an analgesic and an antibiotic.

  In some embodiments, the pharmaceutically active ingredient can be further associated with a polymer, metal, charged compound or charged particle to form a complex thereof.

  In some embodiments, the polymer may include polyglutamic acid (PGA) or other suitable polymer such as hyaluronic acid, chitosan, alginate or dextran.

  In some embodiments, the metal can include zinc or other suitable metals such as calcium, magnesium or iron.

  In some embodiments, the size of the complex is from about 10 nm to about 100 μm.

  In some embodiments, the concentration of the pharmaceutically active ingredient or complex is from about 1 mg / mL to about 300 mg / mL.

  Poly (γ-glutamic acid) (γ-PGA) is a high molecular weight polypeptide composed of a γ-bonded glutamic acid unit and an α-carboxylate side chain. γ-PGA has many advantages, such as non-toxicity, hydrophilicity, biodegradability, and avoidance of antigenicity or immunogenicity, making it an ideal biomaterial that can be applied to create functionalized hydrogel systems It is. In addition, abundant carboxyl groups on the γ-PGA chain can be chemically modified and can bind to soluble antibody molecules by electrostatic interactions.

  Poly (ethylene glycol) (PEG) as an uncharged hydrophilic segment is widely used to build chemically or physically cross-linked hydrogel systems because of its biocompatibility. PEG-based hydrogels have been utilized as cell scaffolds, adhesives for medical applications, and delivery vehicles. In particular, the ability to adjust the crosslink density provides flexibility and adaptability to PEG-based hydrogels and can be used for cell encapsulation and tissue growth.

  In this disclosure, in order to achieve high drug loading (greater than 150 mg / mL) and long-term sustained release (greater than 21 days), subcutaneously injectable and in Develop a situ-formed hydrogel system. γ-PGA is partially modified with N- (2-aminoethyl) maleimide by aminolization. The resulting maleimide-containing γ-PGA (γ-PGA-MA) and 4-arm PEG-SH are mixed in an aqueous solution. A chemically crosslinked hydrogel is formed through a pH-sensitive Michael addition reaction between maleimide and thiol groups.

Example / Comparative Example 1
Preparation of hydrogel composition (1) First, 0.002 g of polyglutamic acid (PGA) (Mw: 1000 kDa, DS (graft ratio): 31.6%) containing a maleimide group was dissolved in 50 μL of PBS, (Concentration: 4.0 wt%). Next, 0.002 g of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group was dissolved in 50 μL of PBS to make a second solution (concentration: 4.0 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 0.4, and the pH of the mixed solution was 5.5. According to visual confirmation, gelation was successful.

Example 2
Preparation of hydrogel composition (2) First, 0.002 g of polyglutamic acid (PGA) (Mw: 1000 kDa, DS (graft ratio): 31.6%) containing a maleimide group was dissolved in 50 μL of PBS, (Concentration: 4.0 wt%). Next, 0.004 g of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group was dissolved in 50 μL of PBS to make a second solution (concentration: 8.0 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 0.8, and the pH of the mixed solution was 5.5. According to visual confirmation, gelation was successful.

Example 3
Preparation of hydrogel composition (3) First, 0.002 g of polyglutamic acid (PGA) (Mw: 1000 kDa, DS (graft ratio): 31.6%) containing a maleimide group was dissolved in 50 μL of PBS, (Concentration: 4.0 wt%). Next, 0.005 g of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group was dissolved in 50 μL of PBS to make a second solution (concentration: 10 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 1.0, and the pH of the mixed solution was 5.5. According to visual confirmation, gelation was successful.

Example 4
Preparation of hydrogel composition (4) First, 0.001 g of polyglutamic acid (PGA) (Mw: 1000 kDa, DS (graft ratio): 31.6%) containing a maleimide group was dissolved in 50 μL of PBS, (Concentration: 2.0 wt%, pH: 4.0). Next, 0.002 g of polyethylene glycol (PEG) containing thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in 50 μL of PBS to make a second solution (concentration: 4.0 wt%, pH : 7.2). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 0.8, and the pH of the mixed solution was 5.6. According to visual confirmation, the gelation was successful and the gelation took 12 minutes.

Example 5
Preparation of hydrogel composition (5) First, 0.001 g of polyglutamic acid (PGA) (Mw: 1000 kDa, DS (graft ratio): 31.6%) containing a maleimide group was dissolved in 50 μL of PBS, (Concentration: 2.0 wt%, pH: 4.0). Next, 0.002 g of polyethylene glycol (PEG) containing thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in 50 μL of deionized water to make a second solution (concentration: 4.0 wt%). , PH: 4.4). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 0.8, and the pH of the mixed solution was 4.3. According to visual confirmation, the gelation was successful and the gelation took 20 minutes.

Example 6
Preparation of hydrogel composition (6) First, 0.001 g of polyglutamic acid (PGA) (Mw: 1000 kDa, DS (graft ratio): 31.6%) containing a maleimide group was dissolved in 50 μL of deionized water, A first solution was made (concentration: 2.0 wt%, pH: 3.9). Next, 0.002 g of polyethylene glycol (PEG) containing thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in 50 μL of deionized water to make a second solution (concentration: 4.0 wt%). , PH: 4.4). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 0.8, and the pH of the mixed solution was 4.1. According to visual confirmation, the gelation was successful and the gelation took more than 6 hours.

Example 7
Preparation of hydrogel composition (7) First, 0.002 g of polyglutamic acid (PGA) (Mw: 1000 kDa, DS (graft ratio): 31.6%) containing a maleimide group was dissolved in 50 μL of PBS, (Concentration: 4.0 wt%, pH: 3.9). Next, 0.002 g of polyethylene glycol (PEG) containing thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in 50 μL of deionized water to make a second solution (concentration: 4.0 wt%). , PH: 4.4). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 0.4, and the pH of the mixed solution was 4.2. According to visual confirmation, the gelation was successful and the gelation took 10 minutes.

Example 8
Preparation of hydrogel composition (8) First, 0.001 g of polyglutamic acid (PGA) (Mw: 1000 kDa, DS (graft ratio): 31.6%) containing a maleimide group was dissolved in 50 μL of PBS, (Concentration: 2.0 wt%, pH: 4.0). Next, 0.002 g of polyethylene glycol (PEG) (Mw: 10 kDa, 4 arm type) containing a thiol group was dissolved in 50 μL of PBS to make a second solution (concentration: 4.0 wt%, pH : 7.2). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 0.4, and the pH of the mixed solution was 5.6. According to visual confirmation, the gelation was successful and the gelation took 2-3 minutes.

Example 9
Preparation of hydrogel composition (9) First, 0.001 g of polyglutamic acid (PGA) (Mw: 1000 kDa, DS (graft ratio): 31.6%) containing a maleimide group was dissolved in 50 μL of PBS, (Concentration: 2.0 wt%, pH: 4.0). Next, 0.0015 g of polyethylene glycol (PEG) containing thiol groups (Mw: 10 kDa, 4 arm type) was dissolved in 50 μL of PBS to make a second solution (concentration: 3 wt%, pH: 7 .2). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 0.3, and the pH of the mixed solution was 5.6. According to visual confirmation, gelation was successful and it took 6-8 minutes to gel.

Example 10
Preparation of Hydrogel Composition (10) First, 0.002 g of polyglutamic acid (PGA) containing maleimide groups (Mw: 200 to 400 kDa, DS (graft ratio): 11.5%) in 50 μL of 0.9% NaCl solution To make a first solution (concentration: 4.0 wt%, pH: 4.2). Next, 0.004 g of polyethylene glycol (PEG) containing thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in 50 μL of 0.9% NaCl solution to make a second solution (concentration: 8 0.0 wt%, pH: 5.5). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 2.0, and the pH of the mixed solution was 4.9. According to visual confirmation, the gelation was successful and the time taken for the gelation was less than 5 minutes.

Example 11
Preparation of hydrogel composition (11) First, 0.002 g of polyglutamic acid (PGA) (Mw: 200 to 400 kDa, DS (graft ratio): 11.5%) containing maleimide groups in 50 μL of 0.9% NaCl solution To make a first solution (concentration: 4.0 wt%, pH: 4.2). Next, 0.006 g of polyethylene glycol (PEG) containing thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in 50 μL of 0.9% NaCl solution to make a second solution (concentration: 12). 0.0 wt%, pH: 5.5). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 3.0, and the pH of the mixed solution was 4.9. According to visual confirmation, the gelation was successful and the time taken for the gelation was less than 5 minutes.

Example 12
Preparation of hydrogel composition (12) First, 0.002 g of polyglutamic acid (PGA) containing maleimide groups (Mw: 200 to 400 kDa, DS (graft ratio): 11.5%) in 50 μL of 0.9% NaCl solution To make a first solution (concentration: 4.0 wt%, pH: 4.2). Next, 0.01 g of polyethylene glycol (PEG) containing a thiol group (Mw: 5 kDa, 4 arm type) was dissolved in 50 μL of 0.9% NaCl solution to make a second solution (concentration: 20 wt. %, PH: 5.5). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 5.0, and the pH of the mixed solution was 4.9. According to visual confirmation, the gelation was successful and the gelation took more than a day.

Example 13
Preparation of hydrogel composition (13) First, 0.01 g of polyglutamic acid (PGA) (Mw: 200 to 400 kDa, DS (graft ratio): 5.4%) containing maleimide groups in 50 μL of 0.9% NaCl solution To make a first solution (concentration: 20 wt%, pH: 4.2). Next, 0.005 g of polyethylene glycol (PEG) containing thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in 50 μL of 0.9% NaCl solution to make a second solution (concentration: 10 wt. %, PH: 5.5). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 1.0, and the pH of the mixed solution was 4.9. According to visual confirmation, the gelation was successful and the gelation took 2-3 minutes.

Example 14
Preparation of hydrogel composition (14) First, 0.01 g of polyglutamic acid (PGA) (Mw: 200 to 400 kDa, DS (graft ratio): 11.5%) containing a maleimide group was dissolved in 50 μL of PBS. A first solution was made (concentration: 20 wt%, pH: 4.2). Next, 0.01 g of polyethylene glycol (PEG) containing a thiol group (Mw: 5 kDa, 4 arm type) was dissolved in 50 μL of 0.9% NaCl solution to make a second solution (concentration: 20 wt. %, PH: 5.5). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 1.0, and the pH of the mixed solution was 4.9. According to visual confirmation, the gelation was successful and the gelation took 1-2 minutes.

Example 15
Preparation of drug delivery system (1) 0.00106 g of Herceptin powder was dissolved in 0.9% NaCl solution to form a drug solution (concentration: 10.6 mg / mL). Next, 0.001 g of polyglutamic acid (PGA) containing maleimide groups (Mw: 1000 kDa, DS (graft ratio): 12.5%) was dissolved in 50 μL of a drug solution to make a first solution (concentration: 2.0 wt%). Next, 0.001 g of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group was dissolved in 50 μL of a drug solution to prepare a second solution (concentration: 2.0 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 0.89, and the pH of the mixed solution was 6.0. According to visual confirmation, the gelation was successful and the gelation took 4 minutes.

Example 16
Preparation of drug delivery system (2) 0.00106 g of Herceptin powder was dissolved in 0.9% NaCl solution to form a drug solution (concentration: 10.6 mg / mL). Next, 0.001 g of polyglutamic acid (PGA) containing maleimide groups (Mw: 1000 kDa, DS (graft ratio): 12.5%) was dissolved in 50 μL of a drug solution to make a first solution (concentration: 2.0 wt%). Next, 0.00075 g of polyethylene glycol (PEG) containing thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in 50 μL of the drug solution to make a second solution (concentration: 1.5 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 0.67, and the pH of the mixed solution was 6.0. According to visual confirmation, the gelation was successful and the gelation took 5 minutes.

Example 17
Preparation of drug delivery system (3) 0.00106 g of Herceptin powder was dissolved in 0.9% NaCl solution to form a drug solution (concentration: 10.6 mg / mL). Next, 0.0015 g of polyglutamic acid (PGA) containing maleimide groups (Mw: 1000 kDa, DS (graft rate): 5.3%) was dissolved in 50 μL of a drug solution to make a first solution (concentration: 3.0 wt%). Next, 0.001 g of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group was dissolved in 50 μL of a drug solution to prepare a second solution (concentration: 2.0 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 1.33, and the pH of the mixed solution was 6.0. According to visual confirmation, the gelation was successful and the gelation took 5 minutes.

Example 18
Preparation of drug delivery system (4) 0.00106 g of Herceptin powder was dissolved in 0.9% NaCl solution to form a drug solution (concentration: 10.6 mg / mL). Next, 0.0015 g of polyglutamic acid (PGA) containing maleimide groups (Mw: 1000 kDa, DS (graft rate): 5.3%) was dissolved in 50 μL of a drug solution to make a first solution (concentration: 3.0 wt%). Next, 0.00075 g of polyethylene glycol (PEG) containing thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in 50 μL of the drug solution to make a second solution (concentration: 1.5 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 1.0, and the pH of the mixed solution was 6.0. According to visual confirmation, the gelation was successful and the gelation took 6 minutes.

Example 19
Preparation of drug delivery system (5) After dissolving 0.1 g of Herceptin powder in deionized water, dialyzed with sodium chloride aqueous solution (0.9%) at 4 ° C. for 24 hours (MWCO: 10000) A solution was formed. The drug solution was then centrifuged (4000 g, 4 ° C.) and concentrated to 145.5 mg / mL. Next, 0.0015 g of polyglutamic acid (PGA) containing maleimide groups (Mw: 1000 kDa, DS (graft rate): 5.3%) was dissolved in 50 μL of a drug solution to make a first solution (concentration: 3.0 wt%). Next, 0.001 g of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group was dissolved in 50 μL of a drug solution to prepare a second solution (concentration: 2.0 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 1.33, and the pH of the mixed solution was 6.0. According to visual confirmation, the gelation was successful and the gelation took 1 minute.

Example 20
Preparation of drug delivery system (6) After dissolving 0.1 g Herceptin powder in deionized water, dialyzed with sodium chloride aqueous solution (0.9%) at 4 ° C. for 24 hours (MWCO: 10000) A solution was formed. The drug solution was then centrifuged (4000 g, 4 ° C.) and concentrated to 145.5 mg / mL. Next, 0.0015 g of polyglutamic acid (PGA) containing maleimide groups (Mw: 1000 kDa, DS (graft rate): 5.3%) was dissolved in 50 μL of a drug solution to make a first solution (concentration: 3.0 wt%). Next, 0.00075 g of polyethylene glycol (PEG) containing thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in 50 μL of the drug solution to make a second solution (concentration: 1.5 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 1.0, and the pH of the mixed solution was 6.0. According to visual confirmation, the gelation was successful and the gelation took 1 minute.

Example 21
Preparation of drug delivery system (7) After dissolving 0.1 g of Herceptin powder in deionized water, dialyzed with sodium chloride aqueous solution (0.9%) at 4 ° C. for 24 hours (MWCO: 10000) A solution was formed. The drug solution was then centrifuged (4000 g, 4 ° C.) and concentrated to 145.5 mg / mL. Next, 0.00075 g of polyglutamic acid (PGA) containing maleimide groups (Mw: 1000 kDa, DS (graft rate): 12.5%) was dissolved in 50 μL of a drug solution to make a first solution (concentration: 1.5 wt%). Next, 0.0006 g of polyethylene glycol (PEG) containing a thiol group (Mw: 5 kDa, 4 arm type) was dissolved in 50 μL of a drug solution to make a second solution (concentration: 1.2 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 0.71, and the pH of the mixed solution was 6.0. According to visual confirmation, the gelation was successful and the gelation took 1 minute.

Example 22
Preparation of drug delivery system (8) After dissolving 0.1 g of Herceptin powder in deionized water, dialyzed with sodium chloride aqueous solution (0.9%) at 4 ° C. for 24 hours (MWCO: 10000) A solution was formed. The drug solution was then centrifuged (4000 g, 4 ° C.) and concentrated to 128.6 mg / mL. Next, 0.00075 g of polyglutamic acid (PGA) containing a maleimide group (Mw: 200 to 400 kDa, DS (graft ratio): 11.3%) was dissolved in 50 μL of a drug solution to form a first solution ( Concentration: 1.5 wt%). Next, 0.00075 g of polyethylene glycol (PEG) containing thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in 50 μL of the drug solution to make a second solution (concentration: 1.5 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 1.0, and the pH of the mixed solution was 6.0. According to visual confirmation, the gelation was successful and the gelation took 20 minutes.

Example 23
Preparation of drug delivery system (9) After dissolving 0.1 g of Herceptin powder in deionized water, dialyzed with sodium chloride aqueous solution (0.9%) at 4 ° C. for 24 hours (MWCO: 10,000) A solution was formed. The drug solution was then centrifuged (4000 g, 4 ° C.) and concentrated to 128.6 mg / mL. Next, 0.001 g of polyglutamic acid (PGA) containing maleimide groups (Mw: 200 to 400 kDa, DS (graft rate): 11.3%) was dissolved in 50 μL of a drug solution to prepare a first solution ( Concentration: 2.0 wt%). Next, 0.001 g of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group was dissolved in 50 μL of a drug solution to prepare a second solution (concentration: 2.0 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 1.0, and the pH of the mixed solution was 6.0. According to visual confirmation, the gelation was successful and the gelation took 10 minutes.

Example 24
Preparation of drug delivery system (10) After dissolving 0.1 g Herceptin powder in deionized water, dialyzed with sodium chloride aqueous solution (0.9%) at 4 ° C. for 24 hours (MWCO: 10,000) A solution was formed. The drug solution was then centrifuged (4000 g, 4 ° C.) and concentrated to 128.6 mg / mL. Next, 0.0015 g of polyglutamic acid (PGA) containing maleimide groups (Mw: 200 to 400 kDa, DS (graft rate): 11.3%) was dissolved in 50 μL of a drug solution to form a first solution ( Concentration: 3.0 wt%). Next, 0.0015 g of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group was dissolved in 50 μL of a drug solution to make a second solution (concentration: 3.0 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 1.0, and the pH of the mixed solution was 6.0. According to visual confirmation, the gelation was successful and the gelation took 4 minutes.

Example 25
Preparation of drug delivery system (11) After dissolving 0.1 g of Herceptin powder in deionized water, it was dialyzed with an aqueous sodium chloride solution (0.9%) at 4 ° C. for 24 hours (MWCO: 10,000). A solution was formed. The drug solution was then centrifuged (4000 g, 4 ° C.) and concentrated to 205 mg / mL. Next, 0.00075 g of polyglutamic acid (PGA) containing a maleimide group (Mw: 200 to 400 kDa, DS (graft ratio): 11.3%) was dissolved in 50 μL of a drug solution to form a first solution ( Concentration: 1.5 wt%). Next, 0.00075 g of polyethylene glycol (PEG) containing thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in 50 μL of the drug solution to make a second solution (concentration: 1.5 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 1.0, and the pH of the mixed solution was 6.0. According to visual confirmation, the gelation was successful and the gelation took 15-17 minutes.

Example 26
Preparation of drug delivery system (12) After dissolving 0.1 g of Herceptin powder in deionized water, dialyzed with sodium chloride aqueous solution (0.9%) at 4 ° C. for 24 hours (MWCO: 10000) A solution was formed. The drug solution was then centrifuged (4000 g, 4 ° C.) and concentrated to 205 mg / mL. Next, 0.001 g of polyglutamic acid (PGA) containing maleimide groups (Mw: 200 to 400 kDa, DS (graft rate): 11.3%) was dissolved in 50 μL of a drug solution to prepare a first solution ( Concentration: 2.0 wt%). Next, 0.001 g of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group was dissolved in 50 μL of a drug solution to prepare a second solution (concentration: 2.0 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 1.0, and the pH of the mixed solution was 6.0. According to visual confirmation, gelation was successful, and it took 5-6 minutes to gel.

Example 27
Preparation of drug delivery system (13) After dissolving 0.1 g of Herceptin powder in deionized water, it was dialyzed with an aqueous sodium chloride solution (0.9%) at 4 ° C. for 24 hours (MWCO: 10,000). A solution was formed. The drug solution was then centrifuged (4000 g, 4 ° C.) and concentrated to 205 mg / mL. Next, 0.0015 g of polyglutamic acid (PGA) containing maleimide groups (Mw: 200 to 400 kDa, DS (graft rate): 11.3%) was dissolved in 50 μL of a drug solution to form a first solution ( Concentration: 3.0 wt%). Next, 0.0015 g of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group was dissolved in 50 μL of a drug solution to make a second solution (concentration: 3.0 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 1.0, and the pH of the mixed solution was 6.0. According to visual confirmation, the gelation was successful and the gelation took 2-3 minutes.

Example 28
Preparation of drug delivery system (14) Herceptin powder 0.1 g was dissolved in deionized water to form a drug solution (concentration: 190 mg / mL). Next, 0.001 g of polyglutamic acid (PGA) containing maleimide groups (Mw: 200 to 400 kDa, DS (graft rate): 11.3%) was dissolved in 50 μL of a drug solution to prepare a first solution ( Concentration: 2.0 wt%). Next, 0.001 g of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group was dissolved in 50 μL of a drug solution to prepare a second solution (concentration: 2.0 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 1.0, and the pH of the mixed solution was 6.0. According to visual confirmation, gelation was successful and it took 3-5 minutes to gel.

Example 29
Preparation of drug delivery system (15) Herceptin powder 0.1 g was dissolved in deionized water to form a drug solution (concentration: 160 mg / mL). Next, 0.00075 g of polyglutamic acid (PGA) containing a maleimide group (Mw: 200 to 400 kDa, DS (graft ratio): 19.8%) was dissolved in 50 μL of a drug solution to form a first solution ( Concentration: 1.5 wt%). Next, 0.0015 g of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group was dissolved in 50 μL of a drug solution to make a second solution (concentration: 3.0 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol groups to maleimide groups was 1.0, and the pH of the mixed solution was 6.0. According to visual confirmation, the gelation was successful and the time taken for the gelation was less than 2 minutes.

Example 30
Preparation of drug delivery system (16) 0.01848 g of Herceptin powder in 50 μL of MOPS buffer and polyglutamic acid (PGA) containing maleimide group in 50 μL of MOPS buffer (Mw: 200 to 400 kDa, DS (graft rate)): 11. 3%) 0.001 g was mixed to form a complex solution, and centrifuged to remove 30 μL of the supernatant. Next, 0.001 g of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group was dissolved in 30 μL of 0.9% NaCl solution and mixed with the complex solution to prepare a hydrogel ( Final drug concentration: 184.8 mg / mL). In this preparation, the molar ratio of thiol groups to maleimide groups was 1.0, and the pH of the mixed solution was 6.0. According to visual confirmation, the gelation was successful and the gelation took 5 minutes.

Example 31
Preparation of drug delivery system (17) A Herceptin / Zn complex solution having a concentration of 128.0 mg / mL was prepared using a 0.9% NaCl solution as a buffer solution. Under acidic conditions with a pH value of 5.0, polyglutamic acid (PGA) containing an appropriate amount of maleimide groups (Mw: 1000 kDa, DS (graft rate): 12.5%), and polyethylene glycol (PEG) containing thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in a Herceptin / Zn complex solution to prepare a Herceptin hydrogel having a PGA concentration of 1.0 wt% and a PEG concentration of 0.75 wt% (molar ratio of thiol groups to maleimide groups) 0.7). Gelation time exceeded 30 minutes.

Example 32
Preparation of drug delivery system (18) A 0.5 M histidine solution was used as a buffer solution to make a Herceptin / Zn complex solution with a concentration of 208.0 mg / mL. Polyacid (PGA) containing an appropriate amount of maleimide groups (Mw: 1000 kDa, DS (graft ratio): 12.5%) and polyethylene glycol (PEG) containing thiol groups under acidic conditions with a pH value of 4.3 (Mw: 5 kDa, 4 arm type) was dissolved in a Herceptin / Zn complex solution to prepare a Herceptin hydrogel having a PGA concentration of 1.3 wt% and a PEG concentration of 0.8 wt% (molar ratio of thiol groups to maleimide groups) Is 0.6). The gel time was 5-10 minutes.

Example 33
Preparation of drug delivery system (19) 0.004 g of doxorubicin powder was dissolved in deionized water to form a drug solution (concentration: 4.0 mg / mL). Next, 0.0015 g of polyglutamic acid (PGA) containing maleimide groups (Mw: 200 to 400 kDa, DS (graft ratio): 11.5%) was dissolved in 50 μL of a drug solution to form a first solution ( Concentration: 3.0 wt%). Next, 0.0015 g of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group was dissolved in 50 μL of a drug solution to make a second solution (concentration: 3.0 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of thiol group to maleimide group was 1.0, and the pH of the mixed solution was 4.5. According to visual confirmation, the gelation was successful and the gelation took 20-30 minutes.

Comparative Example 1
Preparation of drug delivery system 0.002 g of doxorubicin powder was dissolved in deionized water to form a drug solution (concentration: 2.0 mg / mL). Next, 0.001 g of polyglutamic acid (PGA) containing a maleimide group (Mw: 200 to 400 kDa, DS (graft ratio): 11.5%) was dissolved in 50 μL of a drug solution to form a first solution ( Concentration: 2.0 wt%). Next, 0.006 g of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group was dissolved in 50 μL of a drug solution to prepare a second solution (concentration: 12 wt%). Subsequently, the 1st solution and the 2nd solution were mixed by equal volume (50 microliters), and hydrogel was produced. In this preparation, the molar ratio of the thiol group to the maleimide group was 6.0, and the pH of the mixed solution was 5.0. According to visual confirmation, gelation failed.

Example 34
Effect of pH on hydrogel preparation Various solutions of polyglutamic acid (PGA) (Mw: 200-400 kDa, DS (graft rate): 11.5%) containing maleimide groups at a concentration of 1.5 wt% were used with various buffer solutions. Produced. Various solutions of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group with a concentration of 1.5 wt% were prepared using various buffer solutions. A hydrogel was prepared by mixing a solution of polyglutamic acid (PGA) containing a maleimide group and a solution of polyethylene glycol (PEG) containing a thiol group under various pH environments, and the gelation process was observed. Table 1 shows the results such as gel time, homogeneity and hydrogel stiffness.

  In this example, moderate gel time and uniformity of the mixture were obtained at the time of fabrication under an acidic pH environment (ie, pH = 4.2, 4.5, 5.0, and 6.5). Furthermore, hydrogels formed under such conditions have favorable stiffness.

Comparative Example 2
Effect of alkaline pH on encapsulation and dissolution of drug-loaded hydrogels

  A Herceptin solution having a concentration of 50.0 mg / mL and a pH value of 7.8 was prepared. A polyglutamic acid (PGA) (Mw: 1000 kDa, DS (graft rate): 31.6%) containing an appropriate amount of maleimide groups was dissolved in a Herceptin solution to prepare a first solution having a concentration of 2.0 wt%. . Polyethylene glycol (PEG) containing an appropriate amount of thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in Herceptin solution to prepare a second solution having a concentration of 4.0 wt%. The first solution and the second solution were mixed under a pH value of 7.8 (alkaline conditions) (molar ratio of thiol group to maleimide group was 0.72) to prepare a hydrogel (GAEG01). The gel time was less than 2 minutes.

  Next, a polyglutamic acid (PGA) (Mw: 1000 kDa, DS (graft ratio): 31.6%) containing an appropriate amount of maleimide groups is dissolved in a Herceptin solution to obtain a first solution having a concentration of 2.0 wt%. Was made. Polyethylene glycol (PEG) containing an appropriate amount of thiol groups (Mw: 10 kDa, 4 arm type) was dissolved in Herceptin solution to prepare a second solution having a concentration of 4.0 wt%. The first solution and the second solution were mixed under a pH value of 7.8 (alkaline conditions) (molar ratio of thiol group to maleimide group was 0.36) to prepare a hydrogel (GAEG02). The gel time was similarly less than 2 minutes.

  In this comparative example, since the gelation was rapid (less than 2 minutes), the mixture of the first solution and the second solution became non-uniform during the production process. In testing drug release behavior, Herceptin release from the hydrogel was incomplete. Hydrogels (GAEG01 and GAEG02) released Herceptin for only about 28 days. In addition, in drug structure and activity studies, the integrity of Herceptin's molecular structure was severely damaged (formation of numerous Herceptin fragments), and its activity was lost (binding ability to antigen was greatly reduced) ).

Example 35
Effect of Acidic pH on Encapsulation and Dissolution of Drug-Loaded Hydrogel Hydrogel GAEG13 was made according to Example 25. Hydrogel GAEG14 was made according to Example 26. Hydrogel GAEG15 was made according to Example 27. The effect of acidic pH on the encapsulation and dissolution of drug-loaded hydrogels (GAEG13, GAEG14 and GAEG15) was tested.

  In this example, the maximum drug loading concentration of hydrogels (GAEG13, GAEG14 and GAEG15) reached 205 mg / mL. The mixture of the first solution and the second solution (hydrogel / drug) was uniform during the preparation process because the gelation time was sufficient (2-17 minutes). In testing drug release behavior, Herceptin release from the hydrogel was complete. Hydrogels (GAEG13, GAEG14 and GAEG15) released Herceptin for about 50 days (ie, the hydrogel has a sustained release capability). In addition, in drug structure and activity studies, the integrity of the Herceptin molecular structure was maintained, eg, there was still no Herceptin fragment formation on day 35. Therefore, its biological activity was maintained (the ability to bind antigen was still high).

Example 36
Dissolution effect of drug complex-loaded hydrogel (1) 0.01848 g Herceptin powder in 50 μL MOPS buffer and polyglutamic acid (PGA) containing maleimide group in 50 μL MOPS buffer (Mw: 200-400 kDa, DS (graft rate)) (11.3%) was mixed with 0.001 g to form a complex solution, and centrifuged to remove 30 μL of the supernatant. Next, 0.001 g of polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing a thiol group is dissolved in 30 μL of 0.9% NaCl solution, mixed with the complex solution, and hydrogel (PGA01) is prepared. (Final drug concentration 184.8 mg / mL). In this preparation, the molar ratio of thiol groups to maleimide groups was 1.0, and the pH of the mixed solution was 6.0. According to visual confirmation, the gelation was successful and the gelation took 5 minutes.

  In this example, Herceptin release from the hydrogel was complete in testing drug release behavior. Hydrogel (PGA01) released Herceptin for about 42 days (ie, the hydrogel has a sustained release capability). In addition, in drug structure and activity studies, the integrity of Herceptin's molecular structure was maintained, eg, there was still no Herceptin fragment / aggregate formation on day 28. Therefore, its biological activity was maintained (the ability to bind antigen was still high).

Example 37
Dissolution effect of drug complex-loaded hydrogel (2) A Herceptin / Zn complex solution having a concentration of 128.0 mg / mL was prepared using a 0.9% NaCl solution as a buffer solution. Polyglutamic acid (PGA) containing an appropriate amount of maleimide groups (Pw) (Mw: 1000 kDa, DS (graft ratio): 12.5%) and polyethylene glycol (PEG) containing thiol groups under a pH value of 5.0 (acidic conditions) Herceptin (Mw: 5 kDa, 4 arm type) dissolved in Herceptin / Zn complex solution, PGA concentration 1.0 wt%, PEG concentration 0.75 wt% (molar ratio of thiol group to maleimide group is 0.7) A hydrogel was made to produce a hydrogel (GAEGZ001). Gelation time exceeded 30 minutes.

  A 0.5 M histidine solution was used as a buffer solution to prepare a Herceptin / Zn complex solution having a concentration of 208.0 mg / mL. Polyglutamic acid (PGA) containing an appropriate amount of maleimide groups (Mw: 1000 kDa, DS (graft ratio): 12.5%) and polyethylene glycol containing thiol groups (PEG) under a pH value of 4.3 (acidic conditions) ) (Mw: 5 kDa, 4 arm type) dissolved in Herceptin / Zn complex solution, PGA concentration 1.3 wt%, PEG concentration 0.8 wt% (molar ratio of thiol group to maleimide group is 0.6) A Herceptin hydrogel was made to make a hydrogel (GAEGZ002). The gel time was 5-10 minutes.

  In this example, the drug loading concentration of hydrogel (GAEGZ002) was 208 mg / mL. Hydrogel (GAEGZ001) released Herceptin for about 42 days, and hydrogel (GAEGZ002) released Herceptin for up to about 70 days (ie, the hydrogel has a strong sustained release capability). This is because Herceptin and Zn interact to form a chelate, resulting in an enhanced sustained release effect. In addition, in drug structure and activity studies, the integrity and stability of Herceptin's molecular structure was maintained, for example, greater than 90% of the released Herceptin monomer. Therefore, its biological activity was maintained (the ability to bind antigen was still high).

Example 38
Release behavior of drug-loaded hydrogel A Herceptin solution with a concentration of 10.6 mg / mL was prepared using a 0.9% NaCl solution as a buffer solution. Next, polyglutamic acid (PGA) containing an appropriate amount of maleimide groups (PGA) (Mw: 200 to 400 kDa, DS (graft ratio): 11.3%) was dissolved in Herceptin solution to obtain a first concentration of 3.0 wt%. A solution was made. Next, polyethylene glycol (PEG) containing an appropriate amount of thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in Herceptin solution to form a second solution having a concentration of 3.0 wt%. Next, the first solution and the second solution were mixed under a pH value of 6.0 (acidic conditions) (the molar ratio of the thiol group to the maleimide group was 1.0) to prepare a hydrogel. The gel time was 10-15 minutes.

  A Herceptin solution having a concentration of 128.6 mg / mL was prepared using a 0.9% NaCl solution as a buffer solution. Next, polyglutamic acid (PGA) containing an appropriate amount of maleimide groups (PGA) (Mw: 200 to 400 kDa, DS (graft ratio): 11.3%) was dissolved in Herceptin solution to obtain a first concentration of 3.0 wt%. A solution was made. Next, polyethylene glycol (PEG) containing an appropriate amount of thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in Herceptin solution to form a second solution having a concentration of 3.0 wt%. Next, the first solution and the second solution were mixed under a pH value of 6.0 (acidic conditions) (the molar ratio of the thiol group to the maleimide group was 1.0) to prepare a hydrogel (GAEG11). The gel time was 4 minutes.

  A Herceptin solution having a concentration of 205.0 mg / mL was prepared using a 0.9% NaCl solution as a buffer solution. Next, polyglutamic acid (PGA) containing an appropriate amount of maleimide groups (PGA) (Mw: 200 to 400 kDa, DS (graft ratio): 11.3%) was dissolved in Herceptin solution to obtain a first concentration of 3.0 wt%. A solution was made. Next, polyethylene glycol (PEG) containing thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in Herceptin solution to make a second solution having a concentration of 3.0 wt%. Next, the first solution and the second solution were mixed under a pH value of 6.0 (acidic conditions) (the molar ratio of the thiol group to the maleimide group was 1.0) to prepare a hydrogel (GAEG15). The gel time was 4 minutes.

  In this example, in the drug release behavior test, the results show that the higher the drug loading concentration, the longer the drug release period. When the drug loading concentration was 128.6 mg / mL or 205.0 mg / mL, the drug release period was about 42 days.

Example 39
Release behavior of drug complex-loaded hydrogel A 0.5 M histidine solution was used as a buffer solution to prepare a Herceptin / Zn complex solution with a concentration of 9.3 mg / mL. Under a pH value of 4.1 (acid conditions), polyglutamic acid (PGA) containing an appropriate amount of maleimide groups (Mw: 200 to 400 kDa, DS (grafting rate: 9.0%)) and polyethylene glycol containing thiol groups (PEG) ) (Mw: 5 kDa, 4 arm type) dissolved in Herceptin / Zn complex solution to make Herceptin hydrogel with PGA concentration of 1.5 wt% and PEG concentration of 1.5 wt% (molar ratio of thiol group to maleimide group is 1.0), a hydrogel (GAEGZ007) was prepared, and the gelation time was 50 to 60 minutes.

  Using a 0.5 M histidine solution as a buffer solution, a Herceptin / Zn complex solution having a concentration of 93.0 mg / mL was prepared. Under a pH value of 4.2 (acidic conditions), polyglutamic acid (PGA) containing an appropriate amount of maleimide groups (Mw: 200 to 400 kDa, DS (grafting rate: 9.0%)) and polyethylene glycol containing thiol groups (PEG) ) (Mw: 5 kDa, 4 arm type) was dissolved in a Herceptin / Zn complex solution to prepare a Herceptin hydrogel having a PGA concentration of 1.5 wt% and a PEG concentration of 1.5 wt% (molar ratio of thiol groups to maleimide groups) 1.0) and a hydrogel (GAEGZ008) was prepared, and the gelation time was 35 to 40 minutes.

  A 0.5 M histidine solution was used as a buffer solution to prepare a Herceptin / Zn complex solution having a concentration of 171.7 mg / mL. Under a pH value of 4.3 (acidic conditions), polyglutamic acid (PGA) containing an appropriate amount of maleimide groups (Mw: 200 to 400 kDa, DS (grafting rate: 9.0%)) and polyethylene glycol containing thiol groups (PEG) ) (Mw: 5 kDa, 4 arm type) was dissolved in a Herceptin / Zn complex solution to prepare a Herceptin hydrogel having a PGA concentration of 1.5 wt% and a PEG concentration of 1.5 wt% (molar ratio of thiol groups to maleimide groups) 1.0), a hydrogel (GAEGZ009) was prepared, and the gelation time was 25 to 30 minutes.In this example, in the test of drug release behavior, the results indicate that the higher the drug loading concentration, the higher the drug release. It has been shown that the duration is longer, with drug loading concentrations of 93.0 mg / mL or 171.7 mg / mL. To come, the drug release period was also in about 35 days.

Example 40
Effect of acidic pH on the encapsulation and dissolution of drug-loaded hydrogels An Etanercept solution with a concentration of 141.3 mg / mL was made. A polyglutamic acid (PGA) (Mw: 200 to 400 kDa, DS (graft ratio): 5.0%) containing an appropriate amount of maleimide groups is dissolved in an etanercept solution to form a first solution having a concentration of 4.1 wt%. It was. Polyethylene glycol (PEG) containing an appropriate amount of thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in an etanercept solution to make a second solution having a concentration of 1.5 wt%. The first solution and the second solution were mixed under a pH value of 6.3 (acidic conditions) (molar ratio of thiol group to maleimide group was 0.8) to prepare a hydrogel. The gel time was 90-120 minutes.

  Subsequently, polyglutamic acid (PGA) (Mw: 200 to 400 kDa, DS (graft rate): 5.0%) containing an appropriate amount of maleimide groups is dissolved in the etanercept solution, and a first solution having a concentration of 4.9 wt% made. Polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing an appropriate amount of thiol group was dissolved in an etanercept solution to make a second solution having a concentration of 1.5 wt%. The first solution and the second solution were mixed under a pH value of 6.3 (acidic conditions) (molar ratio of thiol group to maleimide group was 0.67) to prepare a hydrogel. The gel time was 45 to 90 minutes.

  Subsequently, polyglutamic acid (PGA) (Mw: 200 to 400 kDa, DS (graft rate): 5.0%) containing an appropriate amount of maleimide groups is dissolved in the etanercept solution, and the first solution having a concentration of 5.7 wt% made. Polyethylene glycol (PEG) containing an appropriate amount of thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in an etanercept solution to make a second solution having a concentration of 1.5 wt%. The first solution and the second solution were mixed under a pH value of 6.3 (acidic conditions) (molar ratio of thiol group to maleimide group was 0.57) to prepare a hydrogel. The gel time was 30-45 minutes.

  Subsequently, polyglutamic acid (PGA) (Mw: 200 to 400 kDa, DS (graft rate): 5.0%) containing an appropriate amount of maleimide groups is dissolved in the etanercept solution, and a first solution having a concentration of 6.5 wt% made. Polyethylene glycol (PEG) containing an appropriate amount of thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in an etanercept solution to make a second solution having a concentration of 1.5 wt%. The first solution and the second solution were mixed under a pH value of 6.3 (acidic conditions) (molar ratio of thiol group to maleimide group was 0.5) to prepare a hydrogel. The gel time was 30-45 minutes.

  In this example, the maximum drug loading concentration of the hydrogel reached 141.3 mg / mL. Because the gel time was sufficient (30-120 minutes), the mixture of the first and second solutions (hydrogel / drug) was uniform during the preparation process. In testing drug release behavior, Herceptin release from the hydrogel was complete. The hydrogel released etanercept for about 30 days (ie, the hydrogel has a sustained release capability). In addition, the drug structure and activity studies maintained the integrity of the molecular structure of etanercept. Therefore, its biological activity was maintained (the ability to bind antigen was still high).

Example 41
Effect of acidic pH on encapsulation and dissolution of drug-loaded hydrogels HSA (human serum albumin) solutions with a concentration of 100 mg / mL were made. A polyglutamic acid (PGA) (Mw: 200-400 kDa, DS (graft rate): 12.1%) containing an appropriate amount of maleimide groups is dissolved in an HSA solution to form a first solution having a concentration of 2.0 wt%. It was. Polyethylene glycol (PEG) containing an appropriate amount of thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in HSA solution to make a second solution with a concentration of 2.0 wt%. The first solution and the second solution were mixed under a pH value of 5.0 to 6.15 (acidic conditions) (molar ratio of thiol group to maleimide group was 1.0) to prepare hydrogel (A). . The gel time was 8-12 minutes.

  An HSA (human serum albumin) solution having a concentration of 200 mg / mL was prepared. Next, polyglutamic acid (PGA) containing an appropriate amount of maleimide groups (PGA) (Mw: 200 to 400 kDa, DS (graft ratio): 12.1%) is dissolved in the HSA solution, and the first concentration of 2.0 wt% is dissolved. A solution was made. Polyethylene glycol (PEG) containing an appropriate amount of thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in HSA solution to make a second solution with a concentration of 2.0 wt%. The first solution and the second solution were mixed under a pH value of 5.0 to 6.15 (acidic conditions) (the molar ratio of the thiol group to the maleimide group was 1.0) to prepare a hydrogel (B). . The gel time was 2.5-4.2 minutes.

  A 300 mg / mL HSA (human serum albumin) solution was made. Next, polyglutamic acid (PGA) containing an appropriate amount of maleimide groups (PGA) (Mw: 200 to 400 kDa, DS (graft ratio): 12.1%) is dissolved in the HSA solution, and the first concentration of 2.0 wt% is dissolved. A solution was made. Polyethylene glycol (PEG) containing an appropriate amount of thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in HSA solution to make a second solution with a concentration of 2.0 wt%. The first solution and the second solution were mixed under a pH value of 6.15 (acidic conditions) (the molar ratio of thiol groups to maleimide groups was 1.0) to prepare hydrogel (C). The gel time was less than 1 minute.

  In this example, the maximum drug loading concentration of hydrogel (A, B and C) reached 100-300 mg / mL. Since the gelation time was sufficient, the mixture of the first solution and the second solution (hydrogel / drug) was uniform during the preparation process. In addition, in drug structure studies, the integrity and stability of the molecular structure of HSA is maintained, for example, 94.1% and 92% of the HSA monomer released from the hydrogels (A, B, and C), respectively. 0.8% and 92.6%.

Comparative Example 3
Effect of alkaline pH on encapsulation and dissolution of drug-loaded hydrogels HSA (human serum albumin) solutions with a concentration of 50 mg / mL were made. A polyglutamic acid (PGA) (Mw: 200-400 kDa, DS (graft ratio): 12.1%) containing an appropriate amount of maleimide groups is dissolved in an HSA solution to form a first solution having a concentration of 2.0 wt%. It was. Polyethylene glycol (PEG) containing an appropriate amount of thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in HSA solution to make a second solution with a concentration of 2.0 wt%. The first solution and the second solution were mixed under a pH value of 8.18 (alkaline conditions) (the molar ratio of the thiol group to the maleimide group was 1.0) to prepare a hydrogel (D). However, the hydrogel immediately gelled.

  In this comparative example, since the gelation was rapid (that is, the hydrogel immediately gelled), the mixture of the first solution and the second solution became non-uniform during the production process. In addition, in drug structure studies, the integrity of the molecular structure of HSA was severely damaged (formation of multiple HSA fragments). Only 75.8% of the HSA monomer was released from the hydrogel.

Example 42
Effect of acidic pH on encapsulation and dissolution of drug-loaded hydrogel A liraglutide solution with a concentration of 20 mg / mL was made. A polyglutamic acid (PGA) (Mw: 200-400 kDa, DS (graft rate): 12.1%) containing an appropriate amount of maleimide groups is dissolved in a liraglutide solution to form a first solution having a concentration of 2.0 wt%. It was. Polyethylene glycol (PEG) containing an appropriate amount of thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in a liraglutide solution to form a second solution having a concentration of 2.0 wt%. The first solution and the second solution were mixed under a pH value of 5.0 (acidic conditions) (molar ratio of thiol group to maleimide group was 1.0) to prepare hydrogel (A). The gel time was 8-12 minutes.

  A liraglutide solution having a concentration of 20 mg / mL was prepared. Next, polyglutamic acid (PGA) (Mw: 300 kDa, DS (graft rate): 12.1%) containing an appropriate amount of maleimide groups is dissolved in the liraglutide solution to form a first solution having a concentration of 3.0 wt%. It was. Polyethylene glycol (PEG) (Mw: 5 kDa, 4 arm type) containing an appropriate amount of thiol group was dissolved in a liraglutide solution to make a second solution having a concentration of 3.0 wt%. The first solution and the second solution were mixed under a pH value of 5.0 (acidic conditions) (the molar ratio of thiol groups to maleimide groups was 1.0) to prepare hydrogel (B). The gel time was 2.5-4.2 minutes.

  In this example, the gelation time was sufficient (2-12 minutes), so the mixture of the first solution and the second solution (hydrogel / drug) became uniform during the preparation process. In addition, in drug structure studies, the integrity and stability of the molecular structure of liraglutide was maintained, for example, 100% of the liraglutide monomer released from the hydrogels (A and B).

Example 43
Study of in-vivo pharmacokinetics (PK) of drug-loaded hydrogels

  First, a “Herceptin” solution having a concentration of 20.0 mg / mL was prepared and used as a control group.

  Another Herceptin solution with a concentration of 20.0 mg / mL was prepared. Subsequently, polyglutamic acid (PGA) (Mw: 200 to 400 kDa, DS (graft ratio): 11.4%) containing an appropriate amount of maleimide groups is dissolved in Herceptin solution, and a first solution having a concentration of 3.0 wt% made. Next, polyethylene glycol (PEG) containing an appropriate amount of thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in Herceptin solution to make a second solution having a concentration of 3.0 wt%. Next, the first solution and the second solution were mixed under a pH value of 6.0 (acidic conditions) (the molar ratio of thiol groups to maleimide groups was 1.0) to prepare “Herceptin-loaded hydrogel”. The experimental group.

Using rats, pharmacokinetic (PK) experiments of “Herceptin” (control group) and “Herceptin-loaded hydrogel” (experimental group) were conducted. The dose was 50 mg / kg. On the 7th day, 14th day, 21st day, 28th day and 35th day after dosing, the drug concentration in plasma was measured, and a serum concentration-time profile was created. Pharmacokinetic parameters such as T _max , C _max , T _1/2 , AUC _D35 and BA (bioavailability) were obtained from the serum concentration-time profile data. They are shown in Table 2.

The results showed that in Herceptin-loaded hydrogel, its C _max was about 60% of the original Herceptin, its T _max was delayed by 2 times, and its T _1/2 was extended by 2.7 times. Thus, the Herceptin-loaded hydrogel has a sustained release capability.

Example 44
Study of in-vivo pharmacokinetics (PK) of hydrogel loaded with drug complex First, a “Herceptin” solution with a concentration of 20.0 mg / mL was prepared as a control group.

  A Herceptin / Zn complex solution having a concentration of 20.0 mg / mL was prepared using a 0.9% NaCl solution as a buffer solution. Polyglutamic acid (PGA) containing an appropriate amount of maleimide groups (Mw: 200 to 400 kDa, DS (grafting rate: 11.4%)) and polyethylene glycol containing thiol groups (PEG) under a pH value of 5.8 (acidic conditions) ) (Mw: 5 kDa, 4 arm type) was dissolved in Herceptin / Zn complex solution at a PGA concentration of 1.5 wt% and a PEG concentration of 1.5 wt% (molar ratio of thiol group to maleimide group was 1. 0), “Herceptin / Zn complex-loaded hydrogel (Her / Zn-hydrogel)” was prepared as an experimental group, and the gelation time was 40 to 50 minutes.

Rats were used to conduct pharmacokinetic (PK) experiments of “Herceptin” (control group) and “Herceptin / Zn complex-loaded hydrogel (Her / Zn-hydrogel)” (experimental group). The dose was 50 mg / kg. On the 7th day, 14th day, 21st day, 28th day and 35th day after dosing, the drug concentration in plasma was measured, and a serum concentration-time profile was created. Pharmacokinetic parameters such as T _max , C _max , T _1/2 , AUC _D35 and BA (bioavailability) were obtained from the serum concentration-time profile data. They are shown in Table 3 and FIG.

The results show that in the Herceptin / Zn complex loaded hydrogel, its C _max is about 30% of the original Herceptin, its T _max is delayed by 3 times, and its T _1/2 is extended by 4.2 times. It was done. Therefore, the Herceptin / Zn complex-loaded hydrogel has a sustained release capability.

Example 45
In-vivo Pharmacodynamics (PD) Study of Drug-Loaded Hydrogel A “Dulbeccoline Buffered Saline (DPBS)” solution was prepared and used as a control group.

  A “Herceptin” solution having a concentration of 10 mg / mL was prepared and used as the first experimental group.

  Another 10 mg / mL Herceptin solution was prepared. Subsequently, polyglutamic acid (PGA) (Mw: 200 to 400 kDa, DS (graft ratio): 11.5%) containing an appropriate amount of maleimide groups is dissolved in Herceptin solution, and a first solution having a concentration of 3.0 wt% made. Next, polyethylene glycol (PEG) containing an appropriate amount of thiol groups (Mw: 5 kDa, 4 arm type) was dissolved in Herceptin solution to make a second solution having a concentration of 3.0 wt%. Next, the first solution and the second solution were mixed under a pH value of 6.0 (acidic conditions) (the molar ratio of thiol groups to maleimide groups was 1.0) to prepare “Herceptin-loaded hydrogel”. The second experimental group.

Using BT474 breast cancer mice, pharmacodynamic (PD) experiments of “DPBS” (control group), “Herceptin” (first experimental group), and “Herceptin-loaded hydrogel” (second experimental group) were performed. The dose of “Herceptin” and “Herceptin-loaded hydrogel” was 50 mg / kg. Tumor volume (mm ³ ) was measured on a specific day after dosing, and a tumor volume change curve was prepared. Tumor growth inhibition (TGI) was calculated from the tumor volume change curve data. They are shown in Table 4.

  From the results of the change in tumor volume on the 28th day after the administration, it can be seen that Herceptin (dose: 50 mg / kg) administered by subcutaneous injection can suppress tumor growth as compared with DPBS, and its TGI is 52. 7%.

  Herceptin-loaded hydrogel TGI on day 28 after dosing is similar to Herceptin.

  There was no significant change in mouse body weight (BW) during the experiment.

Example 46
In-vivo Pharmacodynamics (PD) Study of Drug Complex-Loaded Hydrogel A “Dulbeccoline Buffered Saline (DPBS)” solution was prepared and used as a control group.

  A 0.9% NaCl solution was used as a buffer solution to prepare a Herceptin / Zn complex solution having a concentration of 10.0 mg / mL. Under a pH value of 5.9 (acidic conditions), polyglutamic acid (PGA) containing an appropriate amount of maleimide groups (Mw: 200 to 400 kDa, DS (grafting rate: 11.5%)) and polyethylene glycol containing thiol groups (PEG) ) (Mw: 5 kDa, 4 arm type) was dissolved in Herceptin / Zn complex solution at a PGA concentration of 1.5 wt% and a PEG concentration of 1.5 wt% (molar ratio of thiol group to maleimide group was 1. 0), “Herceptin / Zn complex-loaded hydrogel (Her / Zn-hydrogel)” was prepared as an experimental group, and the gelation time was about 40 minutes.

BT474 breast cancer mice were used to perform pharmacodynamic (PD) experiments of “DPBS” (control group) and “Herceptin / Zn complex-loaded hydrogel (Her / Zn hydrogel)” (experimental group). The dose of “Herceptin / Zn hydrogel” was 50 mg / kg. Tumor volume (mm ³ ) was measured on a specific day after dosing, and a tumor volume change curve was prepared. Tumor growth inhibition (TGI) was calculated from the tumor volume change curve data. They are shown in Table 5 and in FIG.

  From the result of the tumor volume change on the 28th day after the administration, it was found that the Herceptin / Zn complex-loaded hydrogel was able to significantly suppress tumor growth, and its TGI was calculated to be 121.9%.

  There was no significant change in mouse body weight (BW) during the experiment.

The present disclosure provides a novel hydrogel composition comprising polyglutamic acid (PGA) containing maleimide (MA) groups and polyethylene glycol (PEG) containing thiol (SH) groups. In particular, the acidic pH value range of the hydrogel composition is from about 4.0 to about 6.5. The hydrogel can be applied to drug delivery systems. The hydrogel has the ability to carry high doses of drug, with a maximum drug loading concentration reaching about 300 mg / mL. The hydrogel can modulate the release behavior of the drug, for example, the sustained release period reaches at least 35 days in vitro. In particular, in a drug conjugate loaded hydrogel, its C _max is about 30% of the drug not covered by the hydrogel in vivo, its T _max is delayed by 3 times, and its T _1/2 is 4.2 times Extend. The hydrogel maintains the integrity and stability of the drug's molecular structure and biological activity. In addition, the hydrogel can encapsulate many types of drugs, such as high molecular drugs such as antibodies or proteins or peptides, or hydrophilic or hydrophobic small molecules.

  It will be apparent to those skilled in the art that various modifications and variations can be made to the disclosed embodiments. It is intended that the specification and examples be considered as exemplary only, with a true scope of the invention being indicated by the following claims and their equivalents.

Claims (20)

A hydrogel composition comprising:
Polyglutamic acid (PGA) containing maleimide groups;
Including polyethylene glycol (PEG) containing a terminal thiol group,
Ri pH value 4.0 to 6.5 der of the hydrogel composition,
The molar ratio of the thiol group to the maleimide group of the polyethylene glycol (PEG) containing the terminal thiol group and the polyglutamic acid (PGA) containing the maleimide group is 0.2 to 5.0.
Hydrogel composition.   The hydrogel composition according to claim 1, wherein the polyglutamic acid (PGA) containing the maleimide group has a molecular weight of 10 kDa to 1000 kDa.   The hydrogel composition according to claim 1, wherein a graft ratio of the polyglutamic acid (PGA) containing the maleimide group is 5% to 40%. The hydrogel composition according to any one of claims 1 to 3 , wherein a concentration of the polyglutamic acid (PGA) containing the maleimide group in the hydrogel composition is 0.75 wt% to 10 wt%. The hydrogel composition according to any one of claims 1 to 4, wherein the polyglutamic acid (PGA) containing a maleimide group does not contain a thiol group. The hydrogel composition according to any one of claims 1 to 5 , wherein the molecular weight of polyethylene glycol (PEG) containing the terminal thiol group is 2 kDa to 20 kDa. The hydrogel composition according to any one of claims 1 to 6 , wherein a concentration of polyethylene glycol (PEG) containing the terminal thiol group in the hydrogel composition is 0.75 wt% to 10 wt%. The hydrogel composition according to any one of claims 1 to 7, wherein the polyethylene glycol (PEG) containing a terminal thiol group is a 4-arm type, an 8-arm type, or a Y-shape. The hydrogel composition according to any one of claims 1 to 8, wherein the polyethylene glycol (PEG) containing the terminal thiol group does not contain a maleimide group. The molar ratio of the thiol group to the maleimide group of polyethylene glycol (PEG) containing the terminal thiol group and polyglutamic acid (PGA) containing the maleimide group is 1.0 to 1.5. The hydrogel composition of any one of -9 . The hydrogel composition according to any one of claims 1 to 10 ,
A drug delivery system comprising a pharmaceutically active ingredient encapsulated in the hydrogel composition. The drug delivery system according to claim 11 , wherein the pharmaceutically active ingredient is selected from the group consisting of growth factors, hormones, peptides, proteins, antibodies, hydrophilic small molecules and hydrophobic small molecules. 13. A drug delivery system according to one of claims 11 and 12 , wherein the pharmaceutically active ingredient is selected from the group consisting of intact antibodies and antibody fragments. The drug delivery system according to any one of claims 11 to 13, wherein the pharmaceutically active ingredient is selected from the group consisting of a mouse antibody, a chimeric antibody, a humanized antibody, and a human antibody. The drug delivery system according to one of claims 11 and 12 , wherein the pharmaceutically active ingredient is selected from the group consisting of an antitumor drug, an antipsychotic drug, an analgesic and an antibiotic. 16. The drug delivery system according to any one of claims 11 to 15, wherein the pharmaceutically active ingredient is further associated with a polymer, metal, charged compound or charged particle to form a complex thereof. 17. The drug delivery system of claim 16 , wherein the polymer comprises polyglutamic acid (PGA), hyaluronic acid, chitosan, or dextran. The drug delivery system according to claim 16 , wherein the metal comprises zinc, calcium, magnesium or iron. The drug delivery system according to claim 16 , wherein the size of the complex is 10 nm to 100 μm. 20. The drug delivery system according to any one of claims 11, 16, and 19 , wherein the concentration of the pharmaceutically active ingredient or the complex is 1 mg / mL to 300 mg / mL.

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