Multifunctional polyethylene glycol cross-linking agent and preparation method thereof

Abstract

The invention relates to the technical field of organic synthesis, and discloses a multifunctional polyethylene glycol cross-linking agent and a preparation method thereof. The invention discloses a multifunctional polyethylene glycol cross-linking agent, the structural formula of which is # imgabs0 #, wherein n is equal to 2-12. According to the multifunctional polyethylene glycol cross-linking agent provided by the invention, the polyethylene glycol cross-linking agent is connected to a therapeutic protein / polypeptide medicine, so that the slow release effect of the medicine in an organism can be maintained for about 12 hours, and the multifunctional polyethylene glycol cross-linking agent has a better effect of promoting the slow release of the medicine effect; the preparation method of the multifunctional polyethylene glycol cross-linking agent provided by the invention is simple, the raw material cost is low, and large-scale industrial production can be realized.

Description

Technical Field

[0001] The invention relates to the technical field of organic synthesis, and in particular to a multifunctional polyethylene glycol cross-linking agent and a preparation method thereof. Background Art

[0002] Polyethylene glycol is a high molecular polymer that is non-irritating, slightly bitter, has good water solubility, good compatibility with a variety of organic substances, and has excellent lubricity, moisturizing, dispersibility, and adhesion. Therefore, polyethylene glycol is widely used in cosmetics, pharmaceuticals, chemical fibers, rubber, plastics, papermaking, paints, electroplating, pesticides, metal processing, food processing and other fields.

[0003] Polyethylene glycol cross-linkers refer to a class of compounds that introduce cross-linking structures into polyethylene glycol. Such compounds can couple biologically active molecules such as proteins, antibodies, and amino acids. Therefore, polyethylene glycol cross-linkers play an important role in modifying protein drugs, increasing the half-life of drugs in the body, reducing immunogenicity, increasing drug water solubility, and promoting sustained release of drug efficacy.

[0004] Although polyethylene glycol cross-linkers have been widely used in the field of protein drug modification, they have significant disadvantages such as complex synthesis steps and high raw material costs, which leads to many challenges for existing polyethylene glycol cross-linkers in large-scale industrial production. Based on this, developing a new type of polyethylene glycol cross-linker, simplifying its preparation process, and reducing production costs has become a technical problem that technicians in this field urgently need to solve. Summary of the invention

[0005] The purpose of the present invention is to provide a multifunctional polyethylene glycol cross-linking agent and a preparation method thereof. The preparation method of the multifunctional polyethylene glycol cross-linking agent is simple, the raw material cost is low, and large-scale industrial production can be achieved.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] One of the technical solutions of the present invention:

[0008] A multifunctional polyethylene glycol cross-linking agent, the structural formula of which is:

[0009]

[0010] Among them, n=2~12.

[0011] The second technical solution of the present invention:

[0012] The preparation method of the multifunctional polyethylene glycol cross-linking agent comprises the following steps:

[0013] 1) mixing β-mercaptopropionic acid and 1-propanethiol, and oxidizing them to obtain a compound A containing a disulfide bond;

[0014] 2) mixing the compound A obtained in step 1) with oxalyl chloride, and chlorinating the mixture to obtain compound B;

[0015] 3) mixing the compound B obtained in step 2) with aminopolyethylene glycol carboxylic acid, and amidating the mixture to obtain compound C;

[0016] 4) mixing the compound C obtained in step 3) with oxalyl chloride and chlorinating the mixture to obtain compound D;

[0017] 5) N-hydroxysuccinimide and triethylamine are mixed and dissolved, and then the compound D obtained in step 4) is added and stirred for reaction to obtain the multifunctional polyethylene glycol cross-linking agent.

[0018] Furthermore, in step 1), the molar ratio of β-mercaptopropionic acid to 1-propanethiol is 1:(1.2-1.4).

[0019] Furthermore, in step 1), the oxidation is specifically: using an organic peroxide as a catalyst, oxidizing at 40-50° C. for 1-2 hours.

[0020] Further, in step 1), the molecular formula of the compound A is:

[0021]

[0022] Furthermore, in step 2), the molar ratio of compound A to oxalyl chloride is 1:(1.4-1.6).

[0023] Furthermore, in step 2), the chlorination is specifically carried out by using dichloromethane as solvent and 98% by volume concentrated sulfuric acid as catalyst, and chlorinating at 0-10° C. for 3-5 hours.

[0024] Further, in step 2), the molecular formula of the compound B is:

[0025]

[0026] Furthermore, in step 3), the molar ratio of compound B to aminopolyethylene glycol carboxylic acid is 1:(1.6-1.8).

[0027] Furthermore, in step 3), the amidation is specifically carried out by using dichloromethane as solvent and aluminum chloride as catalyst at 10-20° C. for 1-2 hours.

[0028] Further, in step 3), the molecular formula of the compound C is:

[0029]

[0030] Furthermore, in step 4), the molar ratio of compound C to oxalyl chloride is 1:(1.4-1.6).

[0031] Furthermore, in step 4), the chlorination is specifically carried out by using dichloromethane as solvent and 98% by volume concentrated sulfuric acid as catalyst, and chlorinating at 5-15° C. for 0.5-1.5 h.

[0032] Further, in step 4), the molecular formula of the compound D is:

[0033]

[0034] Furthermore, in step 5), the molar ratio of the compound D, N-hydroxysuccinimide and triethylamine is 1:(1.2-1.4):(1.2-1.4).

[0035] Furthermore, in step 5), the dissolving is specifically: using dichloromethane as solvent and stirring until dissolved.

[0036] Furthermore, in step 5), after the dissolution, the mixture needs to be left to stand at room temperature for 30 to 60 minutes.

[0037] Furthermore, in step 5), the stirring reaction is specifically: controlling the stirring speed to 60-90 r / min at room temperature, and stirring the reaction for 3-5 hours.

[0038] The preparation formula of the multifunctional polyethylene glycol cross-linking agent of the present invention is as follows:

[0039]

[0040] The third technical solution of the present invention:

[0041] Application of the above multifunctional polyethylene glycol cross-linking agent in the pharmaceutical field.

[0042] Compared with the prior art, the present invention has the following beneficial effects:

[0043] The multifunctional polyethylene glycol cross-linking agent provided by the present invention can maintain a sustained release effect of the drug in the body for about 12 hours by connecting the polyethylene glycol cross-linking agent to a therapeutic protein / peptide drug, and has a better effect of promoting sustained release of drug efficacy;

[0044] The multifunctional polyethylene glycol cross-linking agent provided by the present invention can improve the physicochemical properties of the drug molecule by connecting the polyethylene glycol cross-linking agent to the therapeutic protein / peptide drug, significantly enhance or change the hydrophilicity, volume, molecular weight, spatial conformation, etc. of the drug molecule, thereby reducing immunogenicity, prolonging the half-life, changing tissue distribution, and increasing the targeting concentration;

[0045] The preparation method of the multifunctional polyethylene glycol cross-linking agent provided by the invention is simple, the raw material cost is low, and large-scale industrial production can be realized. DETAILED DESCRIPTION

[0046] Now, various exemplary embodiments of the present invention are described in detail, and this detailed description should not be considered as a limitation of the present invention, but should be understood as a more detailed description of certain aspects, characteristics and embodiments of the present invention. It should be understood that the terms described in the present invention are only for describing specific embodiments and are not used to limit the present invention.

[0047] In addition, for the numerical range in the present invention, it is understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0048] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0049] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to the skilled artisan. The present invention description and examples are exemplary only.

[0050] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0051] In the following examples and comparative examples, all raw materials used were commercially available unless otherwise specified.

[0052] In the following examples and comparative examples, the room temperature is 25±2° C. unless otherwise specified.

[0053] In the following examples, a method for preparing a multifunctional polyethylene glycol cross-linking agent comprises the following steps:

[0054] 1) adding β-mercaptopropionic acid and 1-propanethiol into a three-necked flask according to the molar ratio of β-mercaptopropionic acid to 1-propanethiol of 1:(1.2-1.4), adding a catalytic amount of an organic peroxide, and oxidizing at 40-50° C. for 1-2 hours to obtain a compound A containing a disulfide bond;

[0055] 2) Compound A and oxalyl chloride obtained in step 1) are added to a three-necked flask at a molar ratio of 1:(1.4-1.6), and concentrated sulfuric acid having a catalytic amount and a volume fraction of 98% is added, and dichloromethane is used as a solvent, and chlorination is carried out at 0-10° C. for 3-5 hours to obtain compound B;

[0056] 3) Compound B and aminopolyethylene glycol carboxylic acid obtained in step 2) are added to a three-necked flask at a molar ratio of 1:(1.6-1.8), and a catalytic amount of aluminum chloride is added, and dichloromethane is used as solvent, and amidation is carried out at 10-20° C. for 1-2 hours to obtain compound C;

[0057] 4) Compound C and oxalyl chloride obtained in step 3) were added to a three-necked flask at a molar ratio of 1:(1.4-1.6), and concentrated sulfuric acid with a catalytic amount volume fraction of 98% was added, and dichloromethane was used as solvent, and chlorination was carried out at 5-15° C. for 0.5-1.5 h to obtain compound D;

[0058] 5) According to the molar ratio of compound D, N-hydroxysuccinimide and triethylamine being 1:(1.2-1.4):(1.2-1.4), N-hydroxysuccinimide and triethylamine are added into a three-necked flask, dichloromethane is used as solvent, and after stirring and dissolving, the mixture is allowed to stand at room temperature for 30-60 min, and then compound D obtained in step 4) is added, and the stirring speed is controlled at 60-90 r / min at room temperature, and the reaction is stirred for 3-5 h to obtain the multifunctional polyethylene glycol crosslinking agent.

[0059] Example 1

[0060] A multifunctional polyethylene glycol cross-linking agent

[0061] 1) adding β-mercaptopropionic acid and 1-propanethiol in a three-necked flask at a molar ratio of 1:1.2, adding a catalytic amount of an organic peroxide, and oxidizing at 40° C. for 1 h to obtain a compound A containing a disulfide bond;

[0062] 2) Compound A and oxalyl chloride obtained in step 1) were added to a three-necked flask at a molar ratio of 1:1.4, and concentrated sulfuric acid with a catalytic amount volume fraction of 98% was added, and dichloromethane was used as solvent, and chlorinated at 0°C for 3h to obtain compound B;

[0063] 3) Compound B and aminopolyethylene glycol carboxylic acid obtained in step 2) were added to a three-necked flask at a molar ratio of 1:1.6, and a catalytic amount of aluminum chloride was added, and dichloromethane was used as solvent, and amidation was carried out at 10° C. for 1 hour to obtain compound C;

[0064] In this embodiment, the chemical formula of the aminopolyethylene glycol carboxylic acid is

[0065] 4) Compound C and oxalyl chloride obtained in step 3) were added to a three-necked flask at a molar ratio of 1:1.4, and concentrated sulfuric acid with a catalytic amount volume fraction of 98% was added, and dichloromethane was used as solvent, and chlorination was carried out at 5°C for 0.5h to obtain compound D;

[0066] 5) According to the molar ratio of compound D, N-hydroxysuccinimide and triethylamine being 1:1.2:1.2, N-hydroxysuccinimide and triethylamine are added into a three-necked flask, dichloromethane is used as solvent, and after stirring and dissolving, the mixture is allowed to stand at room temperature for 30 minutes, and then compound D obtained in step 4) is added, and the stirring speed is controlled at 60 r / min at room temperature, and the reaction is stirred for 3 hours to obtain the multifunctional polyethylene glycol crosslinking agent.

[0067] Example 2

[0068] A multifunctional polyethylene glycol cross-linking agent

[0069] 1) adding β-mercaptopropionic acid and 1-propanethiol in a three-necked flask at a molar ratio of 1:1.3, adding a catalytic amount of an organic peroxide, and oxidizing at 45° C. for 1.5 h to obtain a compound A containing a disulfide bond;

[0070] 2) Compound A and oxalyl chloride obtained in step 1) were added to a three-necked flask at a molar ratio of 1:1.5, and concentrated sulfuric acid with a catalytic amount volume fraction of 98% was added, and dichloromethane was used as solvent, and chlorination was carried out at 5°C for 4 hours to obtain compound B;

[0071] 3) Compound B and aminopolyethylene glycol carboxylic acid obtained in step 2) were added to a three-necked flask at a molar ratio of 1:1.7, and a catalytic amount of aluminum chloride was added, and dichloromethane was used as solvent, and amidation was carried out at 15° C. for 1.5 hours to obtain compound C;

[0072] In this embodiment, the chemical formula of the aminopolyethylene glycol carboxylic acid is

[0073] 4) Compound C and oxalyl chloride obtained in step 3) were added to a three-necked flask at a molar ratio of 1:1.5, and concentrated sulfuric acid with a catalytic amount volume fraction of 98% was added, and dichloromethane was used as solvent, and chlorination was carried out at 10° C. for 1 hour to obtain compound D;

[0074] 5) According to the molar ratio of compound D, N-hydroxysuccinimide and triethylamine being 1:1.3:1.3, N-hydroxysuccinimide and triethylamine are added into a three-necked flask, dichloromethane is used as solvent, and after stirring and dissolving, the mixture is allowed to stand at room temperature for 45 minutes, and then compound D obtained in step 4) is added, and the stirring speed is controlled to 70 r / min at room temperature, and the reaction is stirred for 4 hours to obtain the multifunctional polyethylene glycol crosslinking agent.

[0075] Example 3

[0076] A multifunctional polyethylene glycol cross-linking agent

[0077] 1) adding β-mercaptopropionic acid and 1-propanethiol in a three-necked flask at a molar ratio of β-mercaptopropionic acid to 1-propanethiol of 1:1.4, adding a catalytic amount of an organic peroxide, and oxidizing at 50° C. for 2 h to obtain a compound A containing a disulfide bond;

[0078] 2) Compound A and oxalyl chloride obtained in step 1) were added to a three-necked flask at a molar ratio of 1:1.6, and concentrated sulfuric acid with a catalytic amount volume fraction of 98% was added, and dichloromethane was used as solvent, and chlorinated at 10°C for 5h to obtain compound B;

[0079] 3) Compound B and aminopolyethylene glycol carboxylic acid obtained in step 2) were added to a three-necked flask at a molar ratio of 1:1.8, and a catalytic amount of aluminum chloride was added, and dichloromethane was used as solvent, and amidation was carried out at 20° C. for 2 h to obtain compound C;

[0080] In this embodiment, the chemical formula of the aminopolyethylene glycol carboxylic acid is

[0081] 4) Compound C and oxalyl chloride obtained in step 3) were added to a three-necked flask at a molar ratio of 1:1.6, and concentrated sulfuric acid with a catalytic amount volume fraction of 98% was added, and dichloromethane was used as solvent, and chlorination was carried out at 15° C. for 1.5 hours to obtain compound D;

[0082] 5) According to the molar ratio of compound D, N-hydroxysuccinimide and triethylamine being 1:1.4:1.4, N-hydroxysuccinimide and triethylamine are added into a three-necked flask, dichloromethane is used as solvent, and after stirring and dissolving, the mixture is allowed to stand at room temperature for 60 minutes, and then compound D obtained in step 4) is added, and the stirring speed is controlled to 90 r / min at room temperature, and the reaction is stirred for 5 hours to obtain the multifunctional polyethylene glycol crosslinking agent.

[0083] Embodiments 4 to 8

[0084] A multifunctional polyethylene glycol cross-linking agent

[0085] Same as Example 2, except that: the chemical formulas of the aminopolyethylene glycol carboxylic acids in step 3) are shown in Table 1;

[0086] Table 1 Chemical formula of aminopolyethylene glycol carboxylic acid

[0087]

[0088]

[0089] Effect verification:

[0090] The multifunctional polyethylene glycol cross-linking agent prepared in Examples 1 to 8 was tested for sustained release performance by the following method:

[0091] 1) preparing a 10% bovine serum albumin solution, and then placing the multifunctional polyethylene glycol cross-linking agents prepared in Examples 1 to 8 in the 10% bovine serum albumin solution until swelling equilibrium is reached;

[0092] 2) Weigh 20 mg of the multifunctional polyethylene glycol cross-linking agent that has reached swelling equilibrium and place it in 200 mL of a release medium at a constant temperature (37° C.), sample 5 mL every 1 hour, measure the absorbance at 280 nm using an ultraviolet spectrophotometer, and determine the content of bovine serum albumin in the release medium with reference to the standard curve of bovine serum albumin content, and calculate the cumulative release rate of bovine serum albumin;

[0093] The standard curve of bovine serum albumin content was drawn using conventional drawing methods, which will not be described in detail here;

[0094] After each sampling, the release medium with the same volume as the sample should be added to maintain the total volume unchanged;

[0095] The results of sustained release performance test of the multifunctional polyethylene glycol cross-linking agent prepared in Examples 1 to 8 are shown in Table 2;

[0096] Table 2 Sustained release performance test results

[0097]

[0098]

[0099] It can be seen from the data in Table 2 that the multifunctional polyethylene glycol cross-linking agent provided by the present invention can maintain the sustained-release effect of the drug in the body for about 12 hours, and has an excellent effect of promoting the sustained-release of the drug efficacy.

[0100] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, ordinary technicians in the relevant field should understand that the specific implementation methods of the present invention can still be modified or replaced by equivalents. Any modification or equivalent replacement that does not depart from the spirit and scope of the present invention should be covered within the scope of protection of the claims of the present invention.

Claims

1. A multifunctional polyethylene glycol cross-linking agent, characterized in that The structural formula of the multifunctional polyethylene glycol cross-linking agent is: Among them, n=2~12.

2. A method for preparing the multifunctional polyethylene glycol cross-linking agent according to claim 1, characterized in that: The following steps are involved: 1) mixing β-mercaptopropionic acid and 1-propanethiol, and oxidizing them to obtain a compound A containing a disulfide bond; 2) mixing the compound A obtained in step 1) with oxalyl chloride, and chlorinating the mixture to obtain compound B; 3) mixing the compound B obtained in step 2) with aminopolyethylene glycol carboxylic acid, and amidating the mixture to obtain compound C; 4) mixing the compound C obtained in step 3) with oxalyl chloride and chlorinating the mixture to obtain compound D; 5) N-hydroxysuccinimide and triethylamine are mixed and dissolved, and then the compound D obtained in step 4) is added and stirred for reaction to obtain the multifunctional polyethylene glycol cross-linking agent.

3. The preparation method according to claim 2, characterized in that: In step 1), the molar ratio of β-mercaptopropionic acid to 1-propanethiol is 1:(1.2-1.4); the oxidation is specifically: using an organic peroxide as a catalyst, oxidizing at 40-50° C. for 1-2 hours; the molecular formula of the compound A is:

4. The preparation method according to claim 2, characterized in that: In step 2), the molar ratio of compound A to oxalyl chloride is 1:(1.4-1.6); the chlorination is specifically: using dichloromethane as solvent and 98% by volume concentrated sulfuric acid as catalyst, chlorinating at 0-10°C for 3-5h; the molecular formula of compound B is:

5. The preparation method according to claim 2, characterized in that: In step 3), the molar ratio of compound B to aminopolyethylene glycol carboxylic acid is 1:(1.6-1.8); the amidation is specifically: using dichloromethane as solvent and aluminum chloride as catalyst, amidation is carried out at 10-20° C. for 1-2 hours; the molecular formula of compound C is:

6. The preparation method according to claim 2, characterized in that: In step 4), the molar ratio of the compound C to oxalyl chloride is 1:(1.4-1.6); the chlorination is specifically: using dichloromethane as solvent and 98% concentrated sulfuric acid as catalyst, chlorinating at 5-15°C for 0.5-1.5h; the molecular formula of the compound D is:

7. The preparation method according to claim 2, characterized in that: In step 5), the molar ratio of compound D, N-hydroxysuccinimide and triethylamine is 1:(1.2-1.4):(1.2-1.4); the dissolving is specifically: using dichloromethane as solvent and stirring until dissolved; the stirring reaction is specifically: controlling the stirring speed to 60-90 r / min at room temperature and stirring the reaction for 3-5 hours.

8. The preparation method according to claim 2, characterized in that: In step 5), after the dissolution, the mixture needs to be left to stand at room temperature for 30 to 60 minutes.

9. Use of the multifunctional polyethylene glycol cross-linking agent as claimed in claim 1 in the pharmaceutical field.

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