Preparation method of 2-(2-(2-(3-maleimido-1-oxopropyl) aminoethoxy) ethoxy) acetic acid

Abstract

The invention provides a preparation method of 2-(2-(2-(3-maleimido-1-oxopropyl) amino ethyoxyl) ethyoxyl) acetic acid, which comprises the following step of: reacting 2-(2-(2-amino ethyoxyl) ethyoxyl) acetic acid with 3-maleimide propionic acid-N-hydroxysuccinimide ester in the presence of alkali to obtain the 2-(2-(3-maleimido-1-oxopropyl) amino ethyoxyl) ethyoxyl) acetic acid. And 2-(2-(2-(3-maleimido-1-oxopropyl) amino ethyoxyl) ethyoxyl) acetic acid is generated through the reaction. According to the preparation method provided by the embodiment of the invention, the synthetic route only needs one-step reaction, the route is short, the total yield is high, and the feasibility of the route is stronger.

Description

Technical Field

[0001] The invention belongs to the technical field of compound preparation, and more specifically, relates to a method for preparing 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid. Background Art

[0002] Proteolysis-targeted chimeras (PROTACs) technology is a new drug development strategy that can treat diseases by selectively degrading targeted proteins. Compared with the mechanism of action of traditional drugs, PROTAC technology has many advantages, such as high selectivity, reversibility, and low-dose effect. PROTAC technology has been widely used in many fields, such as cancer, neurodegenerative diseases, immunotherapy, metabolic diseases, infectious diseases, etc.

[0003] PROTAC consists of three parts, including a ligand targeting the target protein, an E3 ligase ligand, and a linker connecting the two ligands. Among them, the linker is a key component of the PROTAC molecule, and its length and composition have a significant impact on the overall physicochemical properties and biological activity. Studies have shown that the introduction of a linker with a polyethylene glycol (PEG) structure can enhance the flexibility and commercial feasibility of the molecule. 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid is a PEG series PROTAC linker. The synthesis method reported in the literature is to first synthesize the corresponding CTC resin and then crack it. This method is cumbersome and not suitable for industrial production. Summary of the invention

[0004] In view of this, the object of the present invention is to provide a method for preparing 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid which is simple to operate, has a short route and is suitable for industrial production.

[0005] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0006] The preparation method of 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid according to an embodiment of the present invention comprises the following steps:

[0007] 2-(2-(2-aminoethoxy)ethoxy)acetic acid is reacted with 3-maleimidopropionic acid-N-hydroxysuccinimide ester in the presence of a base to produce 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid.

[0008] In some embodiments of the present invention, the base is selected from any one or more of sodium hydroxide, sodium carbonate, triethylamine, N,N-diisopropylethylamine, and N-methylmorpholine.

[0009] Preferably, the base is triethylamine.

[0010] In some embodiments of the present invention, the reaction is carried out in a solvent, and the solvent is selected from any one or more of dichloromethane, methanol, ethanol, tetrahydrofuran, N,N-dimethylformamide, and acetonitrile.

[0011] Preferably, the solvent is dichloromethane.

[0012] In some embodiments of the present invention, the preparation method specifically comprises:

[0013] Adding the 2-(2-(2-aminoethoxy)ethoxy)acetic acid and the base to the solvent to dissolve them, thereby obtaining a 2-(2-(2-aminoethoxy)ethoxy)acetic acid solution;

[0014] adding 3-maleimidopropionic acid-N-hydroxysuccinimide ester to the solvent to dissolve it, thereby obtaining a 3-maleimidopropionic acid-N-hydroxysuccinimide ester solution;

[0015] The 3-maleimidopropionic acid-N-hydroxysuccinimide ester solution is added to the 2-(2-(2-aminoethoxy)ethoxy)acetic acid solution, and the mixture is stirred for reaction to generate the 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid.

[0016] In some embodiments of the present invention, the molar ratio of 2-(2-(2-aminoethoxy)ethoxy)acetic acid:3-maleimidopropionic acid-N-hydroxysuccinimide ester:base is 1:(0.8-1.5):(1.0-2.5).

[0017] Preferably, the molar ratio of 2-(2-(2-aminoethoxy)ethoxy)acetic acid:3-maleimidopropionic acid-N-hydroxysuccinimide ester:base is 1:1.0:2.2.

[0018] In some embodiments of the present invention, the reaction temperature is -5 to 40°C; and the reaction time is 3 to 12 hours.

[0019] In some embodiments of the present invention, after the reaction is completed, the following steps are further included:

[0020] Acid is added to make it acidic, then washed with clean water, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain the 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid.

[0021] The above technical solution of the present invention has at least one of the following beneficial effects:

[0022] According to the preparation method of the embodiment of the present invention, the synthesis route only requires one step of reaction, the route is short, the total yield is high, and the feasibility of the route is stronger. DETAILED DESCRIPTION

[0023] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution of the present invention will be clearly and completely described in combination with the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the described embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field belong to the scope of protection of the present invention.

[0024] First, the preparation method of 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid according to an embodiment of the present invention is described in detail below.

[0025] The preparation method of 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid according to an embodiment of the present invention comprises the following steps:

[0026] 2-(2-(2-aminoethoxy)ethoxy)acetic acid (hereinafter, sometimes abbreviated as AEEA) and 3-maleimidopropionic acid-N-hydroxysuccinimide ester (hereinafter, sometimes abbreviated as BMPS) are reacted in the presence of a base to produce 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid.

[0027] Specifically, the reaction formula is shown in the following formula (1):

[0028]

[0029] The synthetic route in the preparation method of the present invention only requires one step of reaction, has a short route, a high total yield, and is more feasible.

[0030] In some embodiments of the present invention, the base is selected from any one or more of sodium hydroxide, sodium carbonate, triethylamine, N,N-diisopropylethylamine (DIPEA), and N-methylmorpholine. Under alkaline conditions, the amino group of AEEA can be exposed, and a weak alkaline environment can be provided for the reaction system, which is conducive to promoting the reaction. Among them, preferably, the base is triethylamine, and the alkalinity of triethylamine can well expose the amino group of AEEA.

[0031] In some embodiments of the present invention, the reaction is carried out in a solvent, and the solvent is selected from any one or more of dichloromethane, methanol, ethanol, tetrahydrofuran, N,N-dimethylformamide (DMF), and acetonitrile. AEEA and BMPS have good solubility in these solvents, which is conducive to the reaction. Among them, dichloromethane is preferred because it is convenient for post-processing.

[0032] In some embodiments of the present invention, the preparation method specifically comprises:

[0033] Adding the 2-(2-(2-aminoethoxy)ethoxy)acetic acid and the base to the solvent to dissolve them, thereby obtaining a 2-(2-(2-aminoethoxy)ethoxy)acetic acid solution;

[0034] adding 3-maleimidopropionic acid-N-hydroxysuccinimide ester to the solvent to dissolve it, thereby obtaining a 3-maleimidopropionic acid-N-hydroxysuccinimide ester solution;

[0035] The 3-maleimidopropionic acid-N-hydroxysuccinimide ester solution is added to the 2-(2-(2-aminoethoxy)ethoxy)acetic acid solution, and the mixture is stirred for reaction to generate the 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid.

[0036] That is, AEEA and BMPS are first dissolved in a solvent to obtain an AEEA solution and a BMPS solution, respectively, and a base is also dissolved in the AEEA solution. Thereafter, the two solutions are mixed and stirred to react to generate the target product 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid. Since a base is added to the AEEA solution in advance, its amino group is exposed, creating a good environment for combining with BMPS during subsequent mixing, thereby facilitating the reaction.

[0037] In some embodiments of the present invention, the molar ratio of 2-(2-(2-aminoethoxy)ethoxy)acetic acid:3-maleimidopropionic acid-N-hydroxysuccinimide ester:base is 1:(0.8-1.5):(1.0-2.5). A slight excess of base can ensure that the amino group of AEEA is completely exposed while providing a weakly alkaline reaction environment. Preferably, the molar ratio of 2-(2-(2-aminoethoxy)ethoxy)acetic acid:3-maleimidopropionic acid-N-hydroxysuccinimide ester:base is 1:1.0:2.2.

[0038] In some embodiments of the present invention, the reaction temperature is -5 to 40°C, and the reaction time is 3 to 12 hours. Preferably, the reaction temperature is 25°C, and the reaction time is 8 hours. The reaction conditions are mild, the equipment requirements are low, the production cost is low, and it is suitable for large-scale industrial production.

[0039] In some embodiments of the present invention, after the reaction is completed, the following steps are further included:

[0040] Acid is added to make it acidic, then washed with clean water, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain the 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid.

[0041] That is, after the reaction is completed, the reaction is quenched by adding acid to acidity, and then the product 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid can be isolated by removing the solvent and water.

[0042] In summary, according to the preparation method of the embodiment of the present invention, the synthetic route only requires one step of reaction, the route is short, the total yield is high, and the feasibility of the route is stronger.

[0043] The following is a detailed description of the preparation method of 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid of the present invention by using specific examples.

[0044] Example 1

[0045] In a 500 mL round-bottom flask, add 100 mL of dichloromethane, 15 g of AEEA and 20.46 g of triethylamine and stir for 20 min;

[0046] 24.47 g BMPS was dissolved in 100 mL of dichloromethane and stirred for 20 min;

[0047] The BMPS solution was added into the AEEA solution, stirred and reacted at room temperature for 8 h.

[0048] Post-treatment: add 2M HCl to adjust to acidity, then wash twice with 80 mL of water, dry over anhydrous sodium sulfate, and evaporate to dryness to obtain 23.26 g of 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid with a yield of 80.5%.

[0049] The reactants were subjected to nuclear magnetic resonance experiments to confirm the product structure. The data are as follows: 1H NMR (400 MHz, d6-DMSO) δ: 12.80 (s, 1H), 8.01 (m, 1H), 7.86 (d, J = 10.9 Hz, 2H), 4.76 (t, J = 7.1 Hz, 2H), 4.31 (s, 2H), 3.67 (t, J = 7.1 Hz, 2H), 3.52 (t, J = 7.1 Hz, 2H), 3.46 (t, J = 7.1 Hz, 2H), 3.28 (t, J = 7.1 Hz, 2H), 2.66 (t, J = 7.1 Hz, 2H). The detection results were consistent with the structure of the target product.

[0050] Example 2

[0051] In a 500 mL round-bottom flask, add 100 mL of dichloromethane, 15 g of AEEA and 20.46 g of N-methylmorpholine and stir for 30 min;

[0052] 24.47 g BMPS was dissolved in 100 mL of dichloromethane and stirred for 30 min;

[0053] The BMPS solution was added into the AEEA solution, stirred and reacted at room temperature for 8 h.

[0054] Post-treatment: add 2M HCl to adjust to acidity, then wash twice with 80 mL of water, dry over anhydrous sodium sulfate, and evaporate to dryness to obtain 20.98 g of 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid with a yield of 72.6%.

[0055] The reactants were subjected to nuclear magnetic resonance experiments to confirm the product structure. The data are as follows: 1H NMR (400 MHz, d6-DMSO) δ: 12.80 (s, 1H), 8.01 (m, 1H), 7.86 (d, J = 10.9 Hz, 2H), 4.76 (t, J = 7.1 Hz, 2H), 4.31 (s, 2H), 3.67 (t, J = 7.1 Hz, 2H), 3.52 (t, J = 7.1 Hz, 2H), 3.46 (t, J = 7.1 Hz, 2H), 3.28 (t, J = 7.1 Hz, 2H), 2.66 (t, J = 7.1 Hz, 2H). The detection results were consistent with the structure of the target product.

[0056] Example 3

[0057] In a 500 mL round-bottom flask, add 80 mL of DMF, 15 g of AEEA and 20.46 g of triethylamine and stir for 20 min;

[0058] Dissolve 24.47 g of BMPS in 80 mL of dichloromethane and stir for 20 min;

[0059] The BMPS solution was added into the AEEA solution, stirred and reacted at room temperature for 8 h.

[0060] Post-treatment: add 2M HCl to adjust to acidity, then wash twice with 80 mL of water, dry over anhydrous sodium sulfate, and evaporate to dryness to obtain 22.42 g of 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid with a yield of 77.6%.

[0061] The reactants were subjected to nuclear magnetic resonance experiments to confirm the product structure. The data are as follows: 1H NMR (400 MHz, d6-DMSO) δ: 12.80 (s, 1H), 8.01 (m, 1H), 7.86 (d, J = 10.9 Hz, 2H), 4.76 (t, J = 7.1 Hz, 2H), 4.31 (s, 2H), 3.67 (t, J = 7.1 Hz, 2H), 3.52 (t, J = 7.1 Hz, 2H), 3.46 (t, J = 7.1 Hz, 2H), 3.28 (t, J = 7.1 Hz, 2H), 2.66 (t, J = 7.1 Hz, 2H). The detection results were consistent with the structure of the target product.

[0062] In summary, the preparation method of 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid of the present invention has the advantages of short synthesis route, simple operation, low cost and high yield, and is suitable for industrial production.

[0063] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for preparing 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid, characterized in that: include: 2-(2-(2-aminoethoxy)ethoxy)acetic acid is reacted with 3-maleimidopropionic acid-N-hydroxysuccinimide ester in the presence of a base to produce 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid.

2. The preparation method according to claim 1, characterized in that: The base is selected from any one or more of sodium hydroxide, sodium carbonate, triethylamine, N,N-diisopropylethylamine, and N-methylmorpholine.

3. The preparation method according to claim 2, characterized in that: The base is triethylamine.

4. The preparation method according to any one of claims 1 to 3, characterized in that: The reaction is carried out in a solvent, and the solvent is selected from any one or more of dichloromethane, methanol, ethanol, tetrahydrofuran, N,N-dimethylformamide, and acetonitrile.

5. The preparation method according to claim 4, characterized in that: The solvent is dichloromethane.

6. The preparation method according to claim 5, characterized in that: The preparation method specifically comprises: Adding the 2-(2-(2-aminoethoxy)ethoxy)acetic acid and the base to the solvent to dissolve them, thereby obtaining a 2-(2-(2-aminoethoxy)ethoxy)acetic acid solution; adding 3-maleimidopropionic acid-N-hydroxysuccinimide ester to the solvent to dissolve it, thereby obtaining a 3-maleimidopropionic acid-N-hydroxysuccinimide ester solution; The 3-maleimidopropionic acid-N-hydroxysuccinimide ester solution is added to the 2-(2-(2-aminoethoxy)ethoxy)acetic acid solution, and the mixture is stirred for reaction to generate the 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid.

7. The preparation method according to claim 1, characterized in that: The molar ratio of 2-(2-(2-aminoethoxy)ethoxy)acetic acid:3-maleimidopropionic acid-N-hydroxysuccinimide ester:base is 1:(0.8-1.5):(1.0-2.5).

8. The preparation method according to claim 7, characterized in that: The molar ratio of 2-(2-(2-aminoethoxy)ethoxy)acetic acid:3-maleimidopropionic acid-N-hydroxysuccinimide ester:base is 1:1.0:2.

2.

9. The preparation method according to claim 1, characterized in that: The reaction temperature is -5 to 40°C; the reaction time is 3 to 12 hours.

10. The preparation method according to claim 1, characterized in that: After the reaction is completed, it also includes: Acid was added to make it acidic, and then washed with clean water, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain the 2-(2-(2-(3-maleimido-1-oxopropyl)aminoethoxy)ethoxy)acetic acid.

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