CN113831529A - Preparation method of monomethoxy polyethylene glycol carboxylic acid and functional derivative thereof - Google Patents
Preparation method of monomethoxy polyethylene glycol carboxylic acid and functional derivative thereof Download PDFInfo
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Abstract
The invention relates to a preparation method of monomethoxy polyethylene glycol carboxylic acid and a functional derivative thereof, which comprises the following steps: (a) removing water from the monomethoxypolyethylene glycol; (b) under the protection of nitrogen, deprotonating monomethoxy polyethylene glycol, and then carrying out Williamson substitution reaction on the deprotonated monomethoxy polyethylene glycol and halogenated carboxylic ester to obtain monomethoxy polyethylene glycol carboxylate; (c) carrying out alkaline hydrolysis on methoxy polyethylene glycol carboxylate, and acidifying to obtain methoxy polyethylene glycol carboxylic acid; (d) under the protection of nitrogen, the methoxy polyethylene glycol carboxylic acid and NHS are subjected to esterification reaction under the dehydration action of DCC to obtain the monomethoxy polyethylene glycol carboxylic acid succinimide ester. The preparation method is simple and convenient, the reaction condition is mild, the product is easy to separate and purify, the comprehensive yield is high, the industrial amplification and commercial application are facilitated, and the prepared methoxy polyethylene glycol carboxylic acid succinimide ester product has high purity and higher commercial application value.
Description
Technical Field
The invention relates to a preparation method of monomethoxy polyethylene glycol carboxylic acid and a functional derivative thereof.
Background
Polyethylene glycol (PEG) technology is a technology for chemically modifying proteins, polypeptides and the like by using polyethylene glycol derivatives. At present, more than ten kinds of protein drugs modified by polyethylene glycol are on the market, amino groups in the protein are mostly randomly modified by polyethylene glycol through modification strategies, and monomethoxypolyethylene glycol active ester is one of common modifying agents. Wherein, the research on the synthetic reaction of the monomethoxy polyethylene glycol carboxylic acid intermediate is the development focus of the monomethoxy polyethylene glycol active ester modifier.
Abuchowski A. et al (Cancer biochem. Biophys.,1984,175-186) report a method of introducing a carboxyl group by esterification of monomethoxypolyethylene glycol with succinic anhydride under basic conditions. However, the modifier obtained by the method is hydrolyzed in vivo after being connected with protein or polypeptide drugs, so that succinic acid remained on the protein or polypeptide serves as a hapten, and the immunogenicity of the protein or polypeptide is enhanced.
Patent document EP0206448 discloses a method of introducing a carboxyl group by oxidation reaction of a hydroxyl group of monomethoxypolyethylene glycol using an oxidizing agent. But the reaction conditions are harsh, the polyethylene glycol chain is easy to break, the reaction byproducts are more, the conversion rate is low, and the method is mainly used for preparing the monomethoxy polyethylene glycol acetic acid.
Patent document US5672662 discloses a method for synthesizing monomethoxy polyethylene glycol propionic acid and monomethoxy polyethylene glycol butyric acid by introducing a cyano group to a hydroxyl end of monomethoxy polyethylene glycol through a nucleophilic addition reaction, performing acidic hydrolysis to monomethoxy polyethylene glycol amide, and then performing basic hydrolysis. But the reaction path is long, the comprehensive yield is low, the industrial amplification is not suitable, the reaction interference factors are more, and the reproducibility of the experimental result is poor.
In order to overcome the defects in the prior art, the application provides a preparation method of monomethoxy polyethylene glycol carboxylic acid and an active derivative thereof.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a synthetic route of monomethoxy polyethylene glycol carboxylic acid, which has mild reaction conditions, high conversion rate and few byproducts, and does not relate to highly toxic chemicals. And further converting the compound into a high-activity modifier, namely monomethoxypolyethylene glycol carboxylic acid succinimide ester.
The invention relates to a preparation method of monomethoxy polyethylene glycol carboxylic acid and methoxy polyethylene glycol carboxylic acid succinimide ester, which comprises the following steps:
(a) removing water from the monomethoxypolyethylene glycol;
(b) under the protection of nitrogen, taking the monomethoxy polyethylene glycol obtained in the step (a) as a raw material, carrying out deprotonation reaction, and carrying out Williamson substitution reaction with halogenated carboxylic ester to obtain monomethoxy polyethylene glycol carboxylate;
(c) carrying out alkaline hydrolysis on the methoxy polyethylene glycol carboxylate obtained in the step (b), and acidifying to obtain methoxy polyethylene glycol carboxylic acid;
(d) and (c) carrying out esterification reaction on the methoxy polyethylene glycol carboxylic acid obtained in the step (c) and N-hydroxysuccinimide (NHS) under the dehydration action of Dicyclohexylcarbodiimide (DCC) under the protection of nitrogen to obtain monomethoxy polyethylene glycol carboxylic acid succinimide ester.
Preferably, in step (a), the monomethoxy polyethylene glycol has a molecular weight of preferably 5000 to 20000Da, and more preferably monomethoxy polyethylene glycol 5000, monomethoxy polyethylene glycol 10000, and monomethoxy polyethylene glycol 20000.
The step (a) specifically comprises the following steps:
1. placing monomethoxy polyethylene glycol and anhydrous toluene in a reaction bottle, fully mixing, connecting a water separator and a condensation pipe, preferably selecting the solid-to-liquid ratio of the monomethoxy polyethylene glycol to the anhydrous toluene to be 1g/10 ml-3 g/10ml, heating the mixed solution under the protection of nitrogen until the mixed solution flows back, keeping the mixed solution in a backflow state for 120-180 min, cooling to room temperature, and discarding the toluene in the water separator. The remaining toluene was evaporated off by a rotary evaporator.
2. Grinding the dried monomethoxy polyethylene glycol, and then drying in vacuum for 12 hours at the temperature of 30-45 ℃ under the vacuum degree of 10-20 mbar.
Step (a) is intended to remove moisture from the monomethoxypolyethylene glycol, and any known method capable of removing moisture from methoxypolyethylene glycol can be used herein without being limited to the method specifically described herein.
Preferably, the step (b) specifically comprises:
1. deprotonation stage of monomethoxypolyethylene glycol:
the bases used for deprotonation of the monomethoxypolyethylene glycol are preferably sodium hydride and potassium tert-butoxide. The dosage of the alkali is different with the difference of the alkali, and when the alkali is sodium hydride, the dosage of the alkali is 20 to 150 times, preferably 100 to 150 times of the molar equivalent of the monomethoxy polyethylene glycol. When potassium tert-butoxide is used as the base, the amount of the base is 5 to 10 times, preferably 10 times, the molar equivalent of the monomethoxypolyethylene glycol. Too little alkali is used, deprotonation is incomplete, substrate conversion is not facilitated, and too much alkali is used, so that the viscosity of a reaction solution is increased, and normal reaction is not facilitated.
The deprotonation temperature is 20-30 ℃. Deprotonation time is 5-24 h. Specific examples are given only by way of example of specific temperatures and times, but the protonation reaction does not greatly affect the reaction results in the above-mentioned temperature and time ranges.
As the reaction solvent, a dry aprotic solvent is used, and toluene, tetrahydrofuran, dichloromethane, N-dimethylformamide and the like are preferable, and toluene and tetrahydrofuran are more preferable. The specific solvent type is determined by the solubility of the deprotonated monomethoxy polyglycol salt in the solvent, and when sodium hydride is used as the base, tetrahydrofuran is most preferred, and when potassium tert-butoxide is used as the base, toluene is most preferred. The monomethoxypolyethylene glycol reaction concentration is preferably 50mg/ml to 150mg/ml, more preferably 75mg/ml to 150 mg/ml.
2. Williamson reaction stage after deprotonation of monomethoxypolyethylene glycol:
the general formula of the halogenated carboxylic ester used in the Williamson reaction is shown as the formula (1):
X-(CH2)m-COO-R(1)
wherein, X is Cl, Br and I, preferably Br and I; m is an integer between 2 and 5; r is a hydrocarbon group having 1 to 6 carbon atoms, preferably methyl, ethyl, propyl, butyl, tert-butyl, pentyl, hexyl.
The amount of the halogenated carboxylic acid ester is preferably 5 to 15 times, more preferably 10 to 15 times, the molar equivalent of the monomethoxy polyethylene glycol.
Williamson reaction time is 48-72 h, the reaction time is too short, the reaction is not thorough, the raw materials are more remained, and the purification difficulty is increased. The reaction temperature is 20-120 ℃, preferably 25-70 ℃. Specific examples are given by way of example only of specific temperatures and times, but do not greatly affect the reaction results within the above-mentioned temperature and time ranges.
Preferably, step (c) specifically comprises:
1. and (3) alkaline hydrolysis reaction, wherein the alkali is inorganic alkali, preferably sodium hydroxide and potassium hydroxide, the concentration of the alkali solution is 0.01-1 mol/l, the hydrolysis reaction temperature is 25-60 ℃, and the reaction time is 6-24 h. Specific examples are only illustrative of specific hydrolysis reaction conditions, but do not greatly affect the reaction results within the above-mentioned ranges.
2. The acid used in the acidification step of the alkaline hydrolysis reaction solution is not particularly limited. The pH value of the reaction liquid after acidification is preferably 2-4, when the pH value is more than 4, the acidity is too weak, the acidification of the carboxylate is incomplete, and when the pH value is less than 2, the acidity is too strong, and the breakage of a polyethylene glycol chain is easy to occur. The acidification reaction temperature is 0-10 ℃. Specific examples are only illustrative of specific acidification reaction conditions, but do not greatly affect the reaction results within the above-mentioned range.
The product obtained is purified by the conventional methods of extraction, salt washing, recrystallization, precipitation, reverse precipitation, chromatography and the like to obtain the methoxypolyethylene glycol carboxylic acid.
Preferably, the step (d) specifically comprises:
in the esterification reaction process, the reaction temperature is 25-30 ℃. The reaction time is 12-24 h. Specific examples are given by way of example only of specific esterification reaction temperatures and times, but do not greatly affect the reaction results within the above-mentioned ranges.
The amount of DCC used is preferably 1 to 3 times, more preferably 1.5 times, the molar equivalent of methoxypolyethylene glycol carboxylic acid. The amount of NHS used is preferably 1 to 3 times, more preferably 1.5 times, the molar equivalent of methoxypolyethylene glycol carboxylic acid.
The reaction solvent is preferably a dry grade solvent, preferably tetrahydrofuran, dichloromethane, most preferably dichloromethane. The monomethoxy polyethylene glycol carboxylic acid reaction concentration is preferably 50mg/ml to 150mg/ml, more preferably 75mg/ml to 125 mg/ml.
The product obtained above is purified by conventional methods such as precipitation, recrystallization, and reprecipitation.
The preparation method of the methoxy polyethylene glycol carboxylic acid and the active derivative thereof provided by the invention is simple and convenient, the reaction product is easy to separate and purify, the separation cost is low, the comprehensive yield is high, the industrial amplification and commercial application are facilitated, the prepared methoxy polyethylene glycol carboxylic acid succinimide ester product is high in purity, and has higher commercial application value, especially the application in the preparation of PEG modified protein or polypeptide drugs.
Drawings
FIG. 1 is a drawing of monomethoxypolyethylene glycol (5000Da) propionic acid of example 11H NMR chart;
FIG. 2 is a drawing of monomethoxypolyethylene glycol (10000Da) propionic acid of example 101H NMR chart;
FIG. 3 is a drawing of monomethoxypolyethylene glycol (20000Da) propionic acid of example 151H NMR chart;
FIG. 4 is a drawing of succinimidyl monomethoxypolyethylene glycol (5000Da) propionate as in example 11H NMR chart;
FIG. 5 is a drawing of succinimidyl monomethoxypolyethylene glycol (10000Da) propionate as in example 101H NMR chart;
FIG. 6 is the preparation of succinimidyl monomethoxypolyethylene glycol (20000Da) propionate of example 151H NMR chart;
FIG. 7 is a GFC chromatogram of monomethoxypolyethylene glycol (5000Da) propionic acid after column separation in example 1;
FIG. 8 is a GFC chromatogram of monomethoxypolyethylene glycol (10000Da) propionic acid after column separation in example 10;
FIG. 9 is a GFC chromatogram of monomethoxypolyethylene glycol (20000Da) propionic acid after column separation in example 15;
FIG. 10 is an HPLC chromatogram of succinimidyl monomethoxypolyethylene glycol (5000Da) propionate of example 1;
FIG. 11 is an HPLC chromatogram of succinimidyl monomethoxypolyethylene glycol (10000Da) propionate from example 10;
FIG. 12 is an HPLC chromatogram of succinimidyl monomethoxypolyethylene glycol (20000Da) propionate of example 15;
Detailed Description
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains, e.g., "PEG" as used herein is commonly referred to as Polyethylene glycol, which is an acronym for Polyethylene glycol, also referred to as "POE," which is a linear polymer with hydroxyl groups at both ends, Polyethylene glycol is polymerized with ethylene oxide, and conventional Polyethylene glycol has a hydroxyl group at each end, and when one end is blocked with a methyl group, monomethoxypolyethylene glycol (mPEG-OH) is obtained. The mPEG-PA is named as Methoxypoly (ethylene glycol) propionic acid and is English abbreviation of monomethoxy polyethylene glycol propionic acid. The mPEG-SPA is called as Methoxypoly (ethylene glycol) succinimidyl propionate, is the English abbreviation of monomethoxypolyethylene glycol succinimidyl propionate, and the derivatives are commonly used in protein pegylation technology.
The embodiment of the invention takes the preparation of methoxypolyethylene glycol propionic acid and methoxypolyethylene glycol propionic acid succinimide ester as an example, and the main reactions are as follows:
the general formula of the methoxypolyethylene glycol propionic acid is shown as a formula (3):
the general formula of the methoxypolyethylene glycol propionic acid succinimide ester is shown as a formula (4):
The synthesis equations presented in the examples are substantially identical to those of formula (2), and the molecular weights of monomethoxypolyethylene glycol were adjusted in the different examples.
The examples of the present invention are only a part of examples of the present invention, and not all examples of the present invention, and based on the examples of the present invention, one skilled in the art can also apply the preparation method to other methoxypolyethylene glycol carboxylic acids (the structural formula is shown in 5), such as methoxypolyethylene glycol butyric acid, valeric acid, etc. And can be further prepared into various polyethylene glycol modifiers, such as methoxy polyethylene glycol carboxylic acid succinimidyl ester (structural formula is shown in 6)
CH3O-(CH2CH2O)n-(CH2)m-COOH (5)
Wherein n is an integer between 112 and 455, and m is an integer between 2 and 5;
CH3O-(CH2CH2O)n-(CH2)m-COO-NHS (6)
wherein n is an integer between 112-455, and m is an integer between 2-5.
Example 1: synthesis of monomethoxypolyethylene glycol (5000Da) propionic acid and active derivative thereof
Adding 2.20g of monomethoxypolyethylene glycol (5000Da) and 20mL of anhydrous toluene into a dry and clean 25mL single-neck bottle, connecting a water separator and a condenser tube, stirring and dissolving under the protection of nitrogen, gradually raising the temperature to reflux, continuing stirring for 2h, cooling to room temperature, performing rotary evaporation to remove residual toluene, and performing vacuum drying for 12 h.
1.60g of sodium hydride (60 percent by weight in kerosene, 40.0mmol) is added into a dry and clean 50mL three-neck flask, 10mL of anhydrous tetrahydrofuran is added under the protection of nitrogen, the mixture is uniformly stirred, 10mL of tetrahydrofuran solution of 2.00g of monomethoxy polyethylene glycol (5000Da, 0.4mmol) obtained in the step is dropwise added, stirring is continued for 4 hours after the dropwise addition, 0.51mL of 3-bromoethyl propionate (4.0mmol) is dropwise added, and the mixture is stirred at room temperature. After TLC monitoring reaction is finished, excess sodium hydride in the system is quenched by absolute ethyl alcohol and saturated ammonium chloride aqueous solution in sequence, reaction liquid is concentrated, dichloromethane extraction residue (30mL x 3) is added, organic phases are combined, drying, concentration and recrystallization are carried out, solid is collected and dried to constant weight, and ethyl monomethoxypolyethylene glycol (5000Da) propionate is obtained.
Adding the ethyl monomethoxypolyethylene glycol propionate obtained in the previous step into a dry and clean 50mL three-necked bottle, adding 20mL of 1mol/L sodium hydroxide aqueous solution, stirring until the ethyl monomethoxypolyethylene glycol propionate is completely dissolved, reacting at 50 ℃ for 12 hours, cooling to room temperature, adding sodium chloride to enable the reaction solution to be in a nearly saturated state, acidifying to pH value of 3 by using 2mol/L hydrochloric acid under an ice salt bath, extracting by dichloromethane (30mL of 3), combining organic phases, drying, concentrating, recrystallizing, collecting solids, and drying to constant weight to obtain a crude monomethoxypolyethylene glycol (5000Da) propionic acid product.
Purifying the crude product obtained in the previous step by an anion exchange column, concentrating the target component collection liquid to 20mL, adding sodium chloride to enable the reaction liquid to be in a nearly saturated state, acidifying the reaction liquid by 2mol/L hydrochloric acid until the pH value is 3, extracting dichloromethane (10mL x 3), combining organic phases, drying, concentrating, recrystallizing, collecting solids, and drying to a constant weight to obtain 1.63g of monomethoxy polyethylene glycol (5000Da) propionic acid, wherein the comprehensive yield is 81.5%, and the sample purity is 97.48%.
Adding the monomethoxy polyethylene glycol (5000Da, 0.33mmol) propionic acid and 56.97mg of N-hydroxysuccinimide (NHS, 0.50mmol) obtained in the previous step into a dry and clean 50mL single-neck bottle, adding 30mL of anhydrous dichloromethane under the protection of nitrogen, stirring until the mixture is dissolved, adding 103.17mg of dicyclohexyl carbodiimide (DCC, 0.50mmol) into 3mL of dichloromethane solution, reacting for 24h at room temperature, filtering to remove insoluble substances, concentrating, recrystallizing, and obtaining 1.44g of monomethoxy polyethylene glycol (5000Da) propionic acid succinimide ester as white powder, wherein the yield is 88.3%, and the sample purity is 98.69%.
The specific parameters and experimental results of examples 1 to 16 are shown in tables 1 to 4, and the steps and parameters not shown in tables 1 to 4 are the same as those in example 1 (equivalent weight of base and beta-haloester is calculated as mPEG-OH; equivalent weight of NHS and DCC is calculated as mPEG-PA intermediate)
TABLE 1 examples 1-9 preparation of monomethoxypolyethylene glycol propionic acids
TABLE 2 examples 1-9 preparation of Monomethoxypolyethylene glycol Propionate succinimidyl ester
TABLE 3 examples 10-16 preparation of monomethoxypolyethylene glycol propionic acids
TABLE 4 examples 10-16 preparation of Monomethoxypolyethylene glycol Propionate succinimidyl ester
Examples 1-16 illustrate the experimental procedure for screening some of the key parameters of the synthetic routes of the present invention. Experiments prove that in examples 1-4, the overall yield of mPEG-PA is increased in a certain trend when the amount of sodium hydride is increased within a certain range, but the overall yield of mPEG-PA is slightly reduced due to the increase of the viscosity of the reaction solution when the amount is increased to 150 eq. Comparative examples 7 and 9 show that potassium tert-butoxide is superior to sodium methoxide in reaction effect.
Comparing examples 6 and 7, it is known that when potassium t-butoxide is used as the base, toluene is selected as the solvent to increase the mPEG-OH conversion. Meanwhile, in comparative examples 15 and 16, it is understood that when sodium hydride is used as the base, tetrahydrofuran is selected as the solvent, and the conversion of mPEG-OH can be improved.
Compared with the examples 1, 2 and 5 and the examples 7 and 8, the yield of the monomethoxy polyethylene glycol propionic acid can be improved to a certain extent by properly increasing the dosage of the beta-halogenated ester, which is favorable for the reaction, but after the dosage of the beta-halogenated ester exceeds 10eq, the dosage is continuously increased, the conversion rate is not increased, and therefore, the dosage of the beta-halogenated ester is preferably about 10 eq. In comparative examples 10-13, the conversion rate of monomethoxy polyethylene glycol propionic acid is increased along with the increase of the mPEG-OH reaction concentration in a certain range, and the reaction concentration of the invention is preferably 75 mg/ml-150 mg/ml.
In the embodiments 1-16 of the present invention, the amount of NHS and DCC is appropriately reduced, preferably 1.1-1.5 eq, dichloromethane is selected as a reaction solvent, the reaction concentration is preferably about 100mg/ml, the monomethoxy polyethylene glycol propionic acid is completely reacted, and the product purity is high. The purities of the target products in fig. 7-12 were 97.48%, 97.12%, 98.95%, 98.69%, 99.27%, 99.91%, respectively.
Example 17: preparation of monomethoxy polyethylene glycol (5000Da) propionic acid and active derivative thereof by amplification process
Adding 63.00g of monomethoxy polyethylene glycol (5000Da) and 600mL of anhydrous toluene into a dry and clean 1L double-layer glass reaction kettle, connecting a water separator and a condenser tube, stirring and dissolving under the protection of nitrogen, gradually raising to reflux, continuously stirring for 2h, cooling to room temperature, performing rotary evaporation to remove residual toluene, and performing vacuum drying for 12 h.
48.00g of sodium hydride (60 percent by weight in kerosene, 1.2mol) is added into a dry and clean 1L double-layer glass reaction kettle, 300mL of anhydrous tetrahydrofuran is added under the protection of nitrogen, the mixture is uniformly stirred, 300mL of tetrahydrofuran solution of 60.00g of monomethoxy polyethylene glycol (5000Da, 12mmol) obtained in the step is dropwise added, the stirring is continued for 8 hours after the dropwise addition, 15.4mL of 3-ethyl bromopropionate (0.12mol) is dropwise added, and the mixture is stirred at room temperature. After TLC monitoring reaction is finished, excess sodium hydride in the system is quenched by absolute ethyl alcohol and saturated ammonium chloride aqueous solution in sequence, reaction liquid is concentrated, dichloromethane extraction residue (300mL x 3) is added, organic phases are combined, drying, concentration and recrystallization are carried out, solid is collected and dried to constant weight, and ethyl monomethoxypolyethylene glycol (5000Da) propionate is obtained.
Adding the ethyl monomethoxypolyethylene glycol propionate obtained in the previous step into a dry and clean 1L three-necked bottle, adding 500mL of 1mol/L sodium hydroxide aqueous solution, stirring until the ethyl monomethoxypolyethylene glycol propionate is completely dissolved, reacting at 50 ℃ for 12 hours, cooling to room temperature, adding sodium chloride to enable the reaction solution to be in a nearly saturated state, acidifying to pH value of 3 by using 2mol/L hydrochloric acid under an ice salt bath, extracting by dichloromethane (300mL of 3), combining organic phases, drying, concentrating, recrystallizing, collecting solids, and drying to constant weight to obtain a crude monomethoxypolyethylene glycol (5000Da) propionic acid product.
Purifying the crude product obtained in the previous step by an anion exchange column, concentrating a target component collection solution to 400mL, adding sodium chloride to enable the reaction solution to be in a nearly saturated state, acidifying the reaction solution by 2mol/L hydrochloric acid until the pH value is 3, extracting dichloromethane (200mL x 3), combining organic phases, drying, concentrating, recrystallizing, collecting solids, drying to a constant weight to obtain 48.18g of monomethoxypolyethylene glycol (5000Da) propionic acid, wherein the comprehensive yield is 80.3%, and the sample purity is 98.11%.
Adding the monomethoxy polyethylene glycol (5000Da, 9.64mmol) propionic acid and 1.66g N-hydroxysuccinimide (NHS, 14.45mmol) obtained in the previous step into a dry and clean 1L single-neck bottle, adding 400mL of anhydrous dichloromethane under the protection of nitrogen, stirring until the mixture is dissolved, adding a 50mL dichloromethane solution of 2.98g dicyclohexyl carbodiimide (DCC, 14.45mmol), reacting for 24h at room temperature, filtering to remove insoluble substances, concentrating, recrystallizing, and obtaining 41.77g of white monomethoxy polyethylene glycol (5000Da) succinimidyl propionate, wherein the yield is 86.7%, and the sample purity is 98.04%.
The embodiment 17 of the invention adopts an amplification process to synthesize the monomethoxy polyethylene glycol (5000Da) propionic acid and the active derivatives thereof, has safe and controllable process and higher product yield and purity, and can be used for the batch preparation of the modifier monomethoxy polyethylene glycol (5000Da) succinimidyl propionate.
Claims (10)
1. The preparation method of the monomethoxy polyethylene glycol carboxylic acid comprises the following steps:
(a) removing water from the monomethoxypolyethylene glycol;
(b) under the protection of nitrogen, taking the monomethoxy polyethylene glycol obtained in the step (a) as a raw material, deprotonating the monomethoxy polyethylene glycol, and carrying out Williamson substitution reaction on the deprotonated monomethoxy polyethylene glycol and halogenated carboxylic ester to obtain monomethoxy polyethylene glycol carboxylic ester;
(c) and (c) carrying out alkaline hydrolysis on the methoxy polyethylene glycol carboxylate obtained in the step (b), and acidifying to obtain methoxy polyethylene glycol carboxylic acid.
2. The process according to claim 1, wherein the base for deprotonation in step (b) is sodium hydride or potassium tert-butoxide; when the alkali is sodium hydride, the dosage of the alkali is 20 to 150 times of the mole equivalent of the monomethoxy polyethylene glycol; when the alkali is potassium tert-butoxide, the amount of the alkali is 5 to 10 times of the mole equivalent of the monomethoxy polyethylene glycol.
3. The method according to claim 2, wherein the monomethoxypolyethylene glycol reaction concentration in the step (b) is 50mg/ml to 150 mg/ml.
4. The process according to claim 1, wherein the deprotonation solvent in step (b) is selected from the group consisting of toluene, tetrahydrofuran, dichloromethane, and N, N-dimethylformamide.
5. The process of claim 1, wherein the halogenated carboxylic acid ester used in the Williamson reaction in step (b) has the formula (1):
X-(CH2)m-COO-R(1)；
wherein X is Cl, Br or I; m is an integer between 2 and 5; r is a hydrocarbyl group having 1 to 6 carbon atoms; the amount of the halogenated carboxylic acid ester is 5 to 15 times of the molar equivalent of the monomethoxy polyethylene glycol.
6. The preparation method of the methoxy polyethylene glycol carboxylic acid succinimide ester comprises the following steps: under the protection of nitrogen, the methoxy polyethylene glycol carboxylic acid and N-hydroxysuccinimide (NHS) are subjected to esterification reaction under the dehydration action of Dicyclohexylcarbodiimide (DCC) to obtain the monomethoxy polyethylene glycol carboxylic acid succinimide ester.
7. The method according to claim 6, wherein the monomethoxypolyethylene glycol carboxylic acid is reacted at a concentration of 50mg/ml to 150 mg/ml.
8. The process according to claim 6, wherein the reaction solvent is tetrahydrofuran or dichloromethane.
9. The method of claim 6, wherein DCC is used in an amount of 1 to 3 times the molar equivalent of methoxypolyethylene glycol carboxylic acid.
10. The method of claim 6, wherein the NHS is used in an amount of 1 to 3 times the molar equivalent of the methoxypolyethylene glycol carboxylic acid.
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