CN114835765B - Synthesis process of 2' -O- (2-methoxyethyl) guanosine - Google Patents
Synthesis process of 2' -O- (2-methoxyethyl) guanosine Download PDFInfo
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Abstract
The invention discloses a synthesis process of 2' -O- (2-methoxyethyl) guanosine, which comprises the following steps: guanosine is reacted with an alkylating agent in a reaction solvent in the presence of a base to produce 2' -O- (2-methoxyethyl) guanosine. The synthetic method of the invention avoids O on the basic group 6 Alkylation improves the product yield, has short preparation steps, high synthesis efficiency, simple and convenient operation, easily obtained raw materials and low requirement on equipment, and is suitable for industrial production.
Description
Technical Field
The invention belongs to the field of chemical synthesis, and particularly relates to a synthesis process of 2' -O- (2-methoxyethyl) guanosine.
Background
Gene therapy is one of the leading technologies at present, and has continuously made breakthrough progress in the treatment of tumors, genetic diseases, metabolic diseases, prevention of infectious diseases, and the like. Among them, the small nucleic acid drug is one of the most rapidly developed gene therapies at present, and is outstanding in the third wave of new drug development. In the future, with continuous breakthrough and innovation in the application field and the technical field of small nucleic acid drugs, the technology updating will contribute to the development of small nucleic acid drugs, and the market demand and the market scale will be continuously expanded.
Antisense oligonucleotides (ASOs) are an emerging class of small nucleic acid drugs with high selectivity and affinity properties. Recently, with the development of the gene group medicine, the antisense oligonucleotide medicine is developed rapidly, with the second generation antisense oligonucleotide medicine deep research, people find in nucleotide introduction 2 '-O-alkyl nucleoside, can enhance with RNA affinity, enhance oligonucleotide to endonuclease and exonuclease resistance, this again with 2' -O-methoxyethyl effect is superior. Therefore, 2' -O- (2-methoxyethyl) guanosine has important market prospect in the field of medicine.
Heretofore, there have been mainly used the following methods for producing 2' -O- (2-methoxyethyl) guanosine.
The preparation method of 2' -O- (2-methoxyethyl) guanosine disclosed in US20080234475 A1 comprises the following reaction equation:
the first step in the process uses tris (2-methoxyethyl) borate as the alkylating agent, the reaction is very water demanding and is a high temperature reaction. The second step of desulfurization by using Raney nickel has higher requirements on production equipment and has great problems on production safety and environmental protection. In addition, the method has the advantages of fewer raw material suppliers, high price and no contribution to industrial production.
2. The preparation method of 2' -O- (2-methoxyethyl) guanosine disclosed in WO 2003087053A 2 has the following reaction equation:
the first step in the process was the use of MDPSCl 2 As protecting group, protecting hydroxyl group at 3 'and 5', reacting in the presence of NaHMDS and TBAI by using 2-bromoethyl methyl ether as alkylating reagent, and deprotecting with tetrabutylammonium fluoride in the third step. The disadvantages of this method are: MDPSCl 2 The protecting group is not yet industrialized and is expensive, so that it cannot be commercially used. And tetrabutylammonium fluoride is difficult to completely remove when purifying the final product.
3. “PROCESS RESEARCH ON THE PREPARATION OF DMT PROTECTED 2'-O-METHOXYETHYLGUANOSINE FOR OLIGONUCLEOTIDE SYNTHESIS IN THERAPEUTIC APPLICATIONS”, Shabbir Ali S. Taj, et al.,Nucleosides, nucleotides and Nucleic Acids, 27 (9): 1024-1033, 2008 discloses a method for preparing 2' -O- (2-methoxyethyl) guanosine, which has the following reaction equation:
in the method, 2-aminoadenosine is used as a starting material to react with 2-bromoethyl methyl ether in the presence of KOH to generate an intermediate product in the first step, and adenosine deaminase is used as the intermediate product to react in a sodium phosphate buffer solution to generate 2' -O- (2-methoxyethyl) guanosine in the second step. The disadvantages of this method are: the conditions of the second step are harsh, and the raw materials are expensive, so that the method is not beneficial to industrial large-scale production.
IN addition, THE above-mentioned prior art "PROCESSS RESONARCH ON THE PREPARTATION OF DMT PROCESSED 2' -O-METHOXYTHYLGLUTANONE FOR OLIGONUCLEOTIDE SYNTHESISS IN THERAPEUTIC APPLICATIONS", shabbir Ali S. Taj,et al.,it is also mentioned in Nucleosides, nucleotides and Nucleic Acids, 27 (9): 1024-1033, 2008 that direct alkylation of guanosine is undesirable because direct alkylation results in partial alkylation of the majority of the product as bases (O) 6 Alkylated) product, which is a by-product, so the document abandons the synthesis of 2'-O- (2-methoxyethyl) guanosine by direct alkylation of 2' -O thereon using guanosine as starting material.
Therefore, the preparation method which is suitable for industrial production, has low requirements on equipment, is easy to obtain raw materials and can obtain high-purity 2' -O- (2-methoxyethyl) guanosine is urgently needed in the field.
Disclosure of Invention
In order to solve the problems in the prior art, the invention aims to provide a synthesis process of 2'-O- (2-methoxyethyl) guanosine, guanosine is used as a raw material to prepare the 2' -O- (2-methoxyethyl) guanosine through one-step reaction, and the synthesis process has the advantages of cheap and easily available raw material, high production efficiency, simple process, easy amplification production and low requirement on equipment. In order to realize the purpose of the invention, the invention adopts the following technical scheme:
a synthesis process of 2' -O- (2-methoxyethyl) guanosine shown in a formula II comprises the following steps: guanosine shown in the formula I and an alkylating reagent react in a reaction solvent in the presence of alkali to generate 2' -O- (2-methoxyethyl) guanosine shown in the formula II, and the reaction equation is as follows:
preferably, in step (I), the molar ratio of guanosine represented by formula I to the alkylating agent is 1:1 to 1:10, more preferably 1:1 to 1:2, most preferably 1 to 1:1.5.
preferably, in step (I), the molar ratio of guanosine represented by formula I to the base is 1:1 to 1:10, more preferably 1:1 to 1:4, most preferably 1 to 1:3.
preferably, the volume molar ratio of the reaction solvent to the guanosine represented by the formula I in the step (I) is 5 to 50L/mol, more preferably 15 to 25L/mol.
Preferably, the reaction temperature in step (i) is from 10 to 100 ℃.
Preferably, the alkylating agent is selected from 2-chloroethyl methyl ether, 2-bromoethyl methyl ether, 2-iodoethyl methyl ether, 2-methoxyethyl methanesulfonate, 2-methoxyethyl p-toluenesulfonate or 2-methoxyethyl p-chlorobenzenesulfonate.
Preferably, when the alkylating agent is selected from 2-chloroethyl methyl ether or 2-bromoethyl methyl ether, the reaction system further comprises an iodine catalyst.
Preferably, the iodine catalyst is selected from one or more of potassium iodide, sodium iodide and tetrabutylammonium iodide.
Preferably, the base is selected from one or more of sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium hydride, butyllithium, sodium tert-butoxide, potassium tert-butoxide and sodium bis (trimethylsilyl) amide.
Preferably, the reaction solvent is selected from one or more of dimethyl sulfoxide, N, N-dimethylformamide, N, N-dimethylacetamide and N-methylpyrrolidone, and more preferably is dimethyl sulfoxide.
Preferably, step (i) further comprises, after the reaction is completed, treating the reaction solution, wherein treating the reaction solution comprises: and cooling the reaction liquid to 5-40 ℃, adding a poor solvent, stirring for 0.5-2 hours, and filtering or centrifuging to obtain a 2' -O- (2-methoxyethyl) guanosine crude product.
Preferably, the above poor solvent is selected from one or more of acetonitrile, propionitrile, butyronitrile, dichloromethane, tetrahydrofuran, methyl tert-butyl ether, acetone and ethyl acetate, more preferably acetonitrile.
Preferably, the volume molar ratio of the poor solvent to the guanosine represented by the formula I is from 30 to 300L/mol, more preferably from 50 to 150L/mol.
Drawings
FIG. 1 is an HPLC chromatogram of 2' -O- (2-methoxyethyl) guanosine obtained in example 1.
FIG. 2 is a HNMR map of 2' -O- (2-methoxyethyl) guanosine obtained in example 1.
FIG. 3 is an HPLC chromatogram of 2' -O- (2-methoxyethyl) guanosine obtained in example 2.
FIG. 4 is an HPLC chromatogram of 2' -O- (2-methoxyethyl) guanosine obtained in example 3.
FIG. 5 is an HPLC chromatogram of 2' -O- (2-methoxyethyl) guanosine obtained in example 4.
FIG. 6 is an HPLC chromatogram of 2' -O- (2-methoxyethyl) guanosine obtained in example 5.
FIG. 7 is an HPLC chromatogram of 2' -O- (2-methoxyethyl) guanosine obtained in example 6.
Detailed Description
Aiming at the defects of the preparation method of 2'-O- (2-methoxyethyl) guanosine in the prior art, the inventor of the application finds that the target product can be prepared by directly carrying out alkylation on 2' -O by taking guanosine as a raw material through deep research, and the method has the advantages of easily available raw materials and simple process, and is suitable for industrial large-scale production. The present invention has been completed on the basis of this finding.
In the invention, the synthesis process of 2' -O- (2-methoxyethyl) guanosine comprises the following steps: guanosine is reacted with an alkylating reagent in a reaction solvent in the presence of a base to form 2' -O- (2-methoxyethyl) guanosine according to the following reaction equation:
in the synthesis method of the present invention, the amount of the alkylating agent may be the amount conventionally used in the art for carrying out such reactions, and preferably, the molar ratio of guanosine represented by the formula I to the alkylating agent is 1:1 to 1:10, more preferably 1:1 to 1:2. the amount of base may be that conventionally used in the art for such reactions, and preferably the molar ratio of guanosine to base represented by formula I is 1:1 to 1:10, more preferably 1:1 to 1:3. the amount of the reaction solvent used is such that the reaction raw material can be completely dissolved at the reaction temperature, and the volume molar ratio of the reaction solvent to the guanosine represented by the formula I is preferably 5 to 50L/mol.
In the synthesis process of the present invention, the alkylating reagent includes, but is not limited to, 2-chloroethyl methyl ether, 2-bromoethyl methyl ether, 2-iodoethyl methyl ether, 2-methoxyethyl methanesulfonate, 2-methoxyethyl p-toluenesulfonate or 2-methoxyethyl p-chlorobenzenesulfonate. Bases include, but are not limited to, sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium hydride, butyllithium, sodium tert-butoxide, potassium tert-butoxide, or sodium bis (trimethylsilyl) amide. The reaction solvent includes, but is not limited to, dimethylsulfoxide, N, N-dimethylformamide, N, N-dimethylacetamide, or N-methylpyrrolidone.
In one embodiment of the present invention, the reaction system of step (i) comprises: guanosine, an alkylating agent selected from 2-bromoethyl methyl ether, a base selected from sodium bis (trimethylsilyl) amide, an iodine catalyst selected from tetrabutylammonium iodide, and a solvent selected from DMF. In one embodiment of the present invention, the process for synthesizing 2' -O- (2-methoxyethyl) guanosine comprises the following steps: guanosine is reacted with 2-bromoethyl methyl ether in a reaction solvent in the presence of sodium bis (trimethylsilyl) amide and tetrabutylammonium iodide at 10 to 40 ℃, more preferably 15 to 35 ℃ for 5 to 7 hours to produce 2' -O- (2-methoxyethyl) guanosine.
In one embodiment of the present invention, the reaction system of step (i) comprises: guanosine, an alkylating agent selected from 2-bromoethyl methyl ether, a base selected from potassium hydroxide, an iodine catalyst selected from potassium iodide, and a solvent selected from DMSO. In one embodiment of the present invention, the process for synthesizing 2' -O- (2-methoxyethyl) guanosine comprises the following steps: guanosine is reacted with 2-bromoethyl methyl ether in a reaction solvent in the presence of potassium hydroxide and potassium iodide at 40 to 80 ℃, preferably 50 to 70 ℃ for 2 to 4 hours to produce 2' -O- (2-methoxyethyl) guanosine.
In one embodiment of the present invention, the reaction system of step (i) comprises: guanosine, an alkylating agent selected from 2-chloroethyl methyl ether, a base selected from potassium hydroxide, an iodine catalyst selected from potassium iodide, and a solvent selected from DMSO. In one embodiment of the present invention, the process for synthesizing 2' -O- (2-methoxyethyl) guanosine comprises the following steps: guanosine is reacted with 2-chloroethyl methyl ether in a reaction solvent in the presence of potassium hydroxide and potassium iodide at 50 to 80 ℃, preferably 60 to 70 ℃ for 15 to 24 hours to produce 2' -O- (2-methoxyethyl) guanosine.
In one embodiment of the present invention, the reaction system of step (i) comprises: guanosine, an alkylating agent selected from 2-iodoethyl methyl ether, a base selected from sodium hydroxide, and a solvent selected from DMSO. In one embodiment of the present invention, the process for synthesizing 2' -O- (2-methoxyethyl) guanosine comprises the following steps: guanosine is reacted with 2-iodoethyl methyl ether in a reaction solvent in the presence of potassium hydroxide and sodium iodide at 10 to 40 ℃, preferably 15 to 25 ℃, for 15 to 24 hours, thereby producing 2' -O- (2-methoxyethyl) guanosine.
In one embodiment of the present invention, the reaction system of step (i) comprises: guanosine, an alkylating agent selected from 2-methoxyethyl methanesulfonate, a base selected from sodium tert-butoxide, and a solvent selected from DMSO. In one embodiment of the present invention, the process for synthesizing 2' -O- (2-methoxyethyl) guanosine comprises the following steps: guanosine is reacted with 2-methoxyethyl methanesulfonate in a reaction solvent in the presence of sodium tert-butoxide at 10 to 40 ℃, more preferably at 15 to 25 ℃ for 15 to 24 hours to produce 2' -O- (2-methoxyethyl) guanosine.
In one embodiment of the present invention, the reaction system of step (i) comprises: guanosine, an alkylating agent selected from 2-methoxyethyl tosylate, a base selected from sodium bis (trimethylsilyl) amide, and a solvent selected from DMF. In one embodiment of the present invention, the process for synthesizing 2' -O- (2-methoxyethyl) guanosine comprises the following steps: guanosine is reacted with 2-methoxyethyl tosylate in a reaction solvent in the presence of sodium bis (trimethylsilyl) amide at 10 to 40 ℃, more preferably 15 to 25 ℃ for 15 to 24 hours to produce 2' -O- (2-methoxyethyl) guanosine.
In the synthesis method of the present invention, whether the reaction is completed or not can be monitored by a method commonly used in the art, for example, TLC, HPLC, or NMR. The time when the raw material guanosine disappears is taken as the reaction end point.
In the synthesis process of the present invention, the reaction temperature depends on the nature of the reagents used to carry out the alkylation. The reaction temperature is 10 to 100 ℃.
In the synthesis method of the present invention, the step (i) further comprises treating the reaction solution after the reaction is completed, wherein the treating the reaction solution comprises: cooling the reaction liquid to 5 to 40 ℃, preferably 15 to 35 ℃, adding a poor solvent, stirring for 0.5 to 2 hours, and filtering or centrifuging to obtain a 2' -O- (2-methoxyethyl) guanosine crude product. The poor solvent includes, but is not limited to, acetonitrile, propionitrile, butyronitrile, dichloromethane, tetrahydrofuran, methyl tert-butyl ether, acetone or ethyl acetate, more preferably acetonitrile. The amount of the poor solvent is such that the product can be sufficiently precipitated from the reaction solvent, and the volume molar ratio of the poor solvent to the guanosine represented by the formula I is preferably from 30 to 300L/mol, and more preferably from 50 to 150L/mol.
In the present invention, the crude 2' -O- (2-methoxyethyl) guanosine can be purified by a method commonly used in the art, such as column chromatography, preferably by a reverse phase column, under the following elution conditions: the mobile phase is acetonitrile and water, wherein the acetonitrile accounts for 5 percent and the water accounts for 95 percent for 30 minutes by volume, so as to obtain the product.
In the synthesis process, the whole system reaction process is a heterogeneous system.
In the description of the present invention, "plural" means two or more.
In the present invention, the reagents used, for example, guanosine, an alkylating agent, a base, an iodine catalyst and the like are commercially available unless otherwise specified.
The synthesis process of 2'-O- (2-methoxyethyl) guanosine takes guanosine as a raw material to synthesize the 2' -O- (2-methoxyethyl) guanosine by one-step reaction, the whole process is simple and convenient, the efficiency is high, the amplification production is easy, the requirement on equipment is not high, the raw material is cheap and easy to obtain, and the use of expensive protecting groups, enzymes and heavy metals is avoided.
The final product yield of the synthetic process of 2'-O- (2-methoxyethyl) guanosine reaches 35 percent, which is close to that of the prior art (the yield of guanosine reported by Nucleotides, nucleotides and Nucleic Acids (2003), 22 (5-8), 1327-1330 is 40 percent when the yield of 2' -O- (2-methoxyethyl) is reported).
The invention will be further illustrated with reference to the following specific examples. The specific embodiment is implemented on the premise of the technical scheme of the invention, and a detailed implementation mode and an operation process are given. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental methods in the following examples, which are not specified under specific conditions, are generally carried out under conventional conditions. Unless otherwise indicated, ratios and percentages are by weight.
The apparatus used in the HPLC assays carried out in the following examples was made by Waters, model UPLC Hclass. The instrument used for the HNMR detection was manufactured as bruker, model avancell 300. The medium pressure chromatography manufacturer performing the purification was orange da, model 1802.
Example 1
Preparation of 2' -O- (2-methoxyethyl) guanosine
Under the protection of nitrogen, 20.0 g of guanosine is taken to be dissolved in 200 mL of DMF at the temperature of between 15 and 30 ℃, 3.8 g of tetrabutylammonium iodide and 17.0 g of 2-bromoethyl methyl ether are added, the temperature is reduced to between 0 and 10 ℃, 106mL of 1.0M sodium bis (trimethylsilyl) amide is dropwise added, after dropwise addition is completed, the temperature is returned to between 15 and 30 ℃, the temperature is kept (between 15 and 30 ℃) for stirring for 6 hours, 2000 mL of ethyl acetate is added, the temperature is kept (between 15 and 30 ℃) for stirring for 1 hour, filtration is carried out, a filter cake is dried in vacuum at the temperature of 50 ℃ to obtain 14.3 g of crude 2' -O-methoxyethyl guanosine, and reverse phase column purification is carried out (the mobile phase comprises acetonitrile and water, wherein the acetonitrile accounts for 5 percent, the water accounts for 95 percent and is kept for 30 ℃ according to volumeMin elution of product) gave 6.8 g of 2' -O- (2-methoxyethyl) guanosine in 28.1% yield and 98.6% HPLC purity. The HPLC profile of the product is shown in FIG. 1, with 0.8 min as the product peak. The HNMR map of the product is shown in figure 2, 1 the H NMR spectrum was assigned in Table 1.
The product has the structural formula shown in formula II:
example 2
Preparation of 2' -O- (2-methoxyethyl) guanosine
Dissolving 10.0 g of guanosine in 200 mL of DMSO (dimethylsulfoxide) at 15 to 30 ℃, adding 10.0 g of potassium hydroxide and 6.0 g of potassium iodide, heating to 50 to 70 ℃, adding 15.0 g of 2-bromoethyl methyl ether, keeping the temperature (50 to 70 ℃) for stirring for 3 hours, cooling to 15 to 30 ℃, adding 1000 mL of dichloromethane, keeping the temperature (15 to 30 ℃) for stirring for 1 hour, filtering, and drying a filter cake in vacuum at 50 ℃ to obtain 8.1 g of a 2'-O-methoxyethyl guanosine crude product, wherein the mobile phase comprises acetonitrile and water, wherein acetonitrile 5% by volume and water 95% by volume are kept for 30 minutes to elute the product), so that 3.8 g of 2' -O- (2-methoxyethyl) guanosine is obtained, the yield is 31.6%, and the purity of HPLC is 98.3%. The HPLC profile of the product is shown in FIG. 3, where 0.8 min is the product peak.
Example 3
Preparation of 2' -O- (2-methoxyethyl) guanosine
Dissolving 100.0 g of guanosine in 1000 mL of DMSO at 15-30 ℃, adding 79.0 g of potassium hydroxide and 58.0 g of potassium iodide, heating to 60-70 ℃, adding 67.1 g of 2-chloroethyl methyl ether, keeping the temperature (60-70 ℃) for stirring for 19 hours, cooling to 15-30 ℃, adding 15000 mL of acetonitrile, keeping the temperature (15-30 ℃) for stirring for 1 hour, filtering, and drying a filter cake at 50 ℃ in vacuum to obtain 93.0 g of a 2'-O-methoxyethyl guanosine crude product, and performing reverse phase column purification (the mobile phase comprises acetonitrile and water, wherein acetonitrile is 5% by volume, water is 95% by volume, and the product is eluted after the vacuum drying is kept for 30 minutes) to obtain 42.4 g of 2' -O- (2-methoxyethyl) guanosine, the yield is 35.2%, and the HPLC purity is 98.3%. The HPLC profile of the product is shown in FIG. 4, where 0.8 min is the product peak.
Example 4
Preparation of 2' -O- (2-methoxyethyl) guanosine
Dissolving 20.0 g of guanosine in 200 mL of DMSO (dimethylsulfoxide) at 15 to 30 ℃, adding 11.3 g of sodium hydroxide, controlling the temperature to 15 to 25 ℃, adding 26.4 g of 2-iodoethyl methyl ether, stirring for 19 hours under the condition of heat preservation (15 to 25 ℃), adding 3000 mL of tetrahydrofuran, stirring for 1 hour under the condition of heat preservation (15 to 25 ℃), filtering, and drying a filter cake under vacuum at 50 ℃ to obtain 19.0 g of crude 2'-O-methoxyethyl guanosine, and performing reverse phase column purification (the mobile phase comprises acetonitrile and water, wherein the acetonitrile is 5% by volume, the water is 95% by volume, and the water is kept for 30 minutes) to obtain 7.4 g of 2' -O- (2-methoxyethyl) guanosine, the yield is 30.6%, and the HPLC purity is 97.2%. The HPLC profile of the product is shown in FIG. 5, with the product peak at 0.8 min.
Example 5
Preparation of 2' -O- (2-methoxyethyl) guanosine
Dissolving 10.0 g of guanosine in 200 mL of DMSO at 15-30 ℃, adding 9.7 g of sodium tert-butoxide, controlling the temperature at 15-25 ℃, adding 12.0 g of methanesulfonic acid-2-methoxyethyl ester, stirring for 19 hours at the temperature of 15-25 ℃, adding 1000 mL of methyl tert-butyl ether, stirring for 1 hour at the temperature of 15-25 ℃, filtering, and drying a filter cake at 50 ℃ in vacuum to obtain 10.5 g of crude 2'-O-methoxyethyl guanosine, and purifying by using an inverse phase column (wherein the mobile phase comprises acetonitrile and water, the volume of the acetonitrile is 5%, the volume of the water is 95%, the content of the water is kept for 30 minutes) to obtain 3.6 g of 2' -O- (2-methoxyethyl) guanosine, the yield is 30.1%, and the HPLC purity is 97.6%. The HPLC profile of the product is shown in FIG. 6, where 0.8 min is the product peak.
Example 6
Preparation of 2' -O- (2-methoxyethyl) guanosine
Dissolving 50.0 g of guanosine in 1000 mL of DMF at 15-30 ℃, cooling to 0-10 ℃, dropwise adding 264mL of 1M sodium bis (trimethylsilyl) amide tetrahydrofuran solution, adding 81.7 g of 2-methoxyethyl p-toluenesulfonate, returning to 15-30 ℃, keeping the temperature (15-30 ℃) for 19 hours while stirring, adding 8000 mL of acetone, keeping the temperature (15-30 ℃) for 1 hour while stirring, filtering, and drying a filter cake at 50 ℃ in vacuum to obtain 51.0 g of crude 2'-O-methoxyethyl guanosine, and purifying by using an inverse phase column (the mobile phase comprises acetonitrile and water, wherein the volume of the acetonitrile is 5 percent, the volume of the water is 95 percent, the content of the water is kept for 30 minutes) to obtain 19.6 g of 2' -O- (2-methoxyethyl) guanosine, the yield is 32.6 percent, and the HPLC purity is 98.2 percent. The HPLC profile of the product is shown in FIG. 7, where 0.8 min is the product peak.
The above embodiments are merely preferred embodiments of the present invention, which are not intended to limit the scope of the present invention, and various changes may be made in the above embodiments of the present invention. All simple and equivalent changes and modifications made according to the claims and the content of the specification of the present application fall within the scope of the claims of the present patent application. The invention has not been described in detail in order to avoid obscuring the invention.
Claims (6)
1. A synthesis process of 2' -O- (2-methoxyethyl) guanosine represented by a formula II, which is characterized by comprising the following steps: guanosine shown in the formula I and an alkylating reagent react in a reaction solvent in the presence of alkali to generate 2' -O- (2-methoxyethyl) guanosine shown in the formula II, wherein the reaction equation is as follows:
the alkylating reagent is selected from 2-chloroethyl methyl ether, 2-bromoethyl methyl ether, 2-iodoethyl methyl ether, methanesulfonic acid-2-methoxyethyl ester, p-toluenesulfonic acid-2-methoxyethyl ester or p-chlorobenzenesulfonic acid-2-methoxyethyl ester,
the alkali is selected from one or more of sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium hydride, butyl lithium, sodium tert-butoxide, potassium tert-butoxide and sodium bis (trimethylsilyl) amide,
when the alkylating agent is selected from 2-chloroethylmethyl ether or 2-bromoethyl methyl ether, the reaction system of step (i) further comprises an iodine catalyst,
the iodine catalyst is selected from one or more of potassium iodide, sodium iodide and tetrabutylammonium iodide.
2. The process of claim 1, wherein step (i) has one or more characteristics selected from the group consisting of:
the molar ratio of guanosine represented by the formula I to the alkylating agent is 1:1 to 1:10, and/or
The molar ratio of guanosine represented by the formula I to the base is 1:1 to 1:10, and/or
The volume molar ratio of the reaction solvent to the guanosine shown in the formula I is 5-50L/mol.
3. The process of claim 1, wherein the reaction solvent is selected from the group consisting of one or more of dimethylsulfoxide, N, N-dimethylformamide, N, N-dimethylacetamide, and N-methylpyrrolidone.
4. The process of claim 1, wherein the reaction system of step (i) comprises:
the alkylating agent selected from 2-bromoethyl methyl ether, the base selected from sodium bis (trimethylsilyl) amide, and an iodine catalyst selected from tetrabutylammonium iodide, or
The alkylating agent is selected from 2-bromoethyl methyl ether, the base is selected from potassium hydroxide, and the iodine catalyst is selected from potassium iodide, or
The alkylating agent is selected from 2-chloroethyl methyl ether, the base is selected from potassium hydroxide, and the iodine catalyst is selected from potassium iodide, or
Said alkylating agent selected from 2-iodoethyl methyl ether, and said base selected from sodium hydroxide, or
The alkylating agent is selected from methanesulfonic acid-2-methoxyethyl ester, and the base is selected from sodium tert-butoxide, or
The alkylating agent selected from 2-methoxyethyl p-toluenesulfonate and the base selected from sodium bis (trimethylsilyl) amide.
5. The process of claim 1, wherein step (i) further comprises treating the reaction solution after the reaction is completed, the treating the reaction solution comprising: and cooling the reaction liquid to 5-40 ℃, adding a poor solvent, stirring for 0.5-2 hours, and filtering or centrifuging to obtain a 2' -O- (2-methoxyethyl) guanosine crude product.
6. The process of claim 5, wherein the poor solvent is selected from one or more of acetonitrile, propionitrile, butyronitrile, dichloromethane, tetrahydrofuran, methyl tert-butyl ether, acetone, and ethyl acetate.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2003087053A2 (en) * | 2002-04-11 | 2003-10-23 | Isis Pharmaceuticals, Inc. | Reagent and process for protecting nucleoside active groups |
| CN102532228A (en) * | 2010-12-31 | 2012-07-04 | 上海兆维科技发展有限公司 | 2'-O-(2-methoxyethyl)adenosine and 2'-O-(2-methoxyethyl)guanosine, preparation of derivatives thereof and purifying methods thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003087053A2 (en) * | 2002-04-11 | 2003-10-23 | Isis Pharmaceuticals, Inc. | Reagent and process for protecting nucleoside active groups |
| CN102532228A (en) * | 2010-12-31 | 2012-07-04 | 上海兆维科技发展有限公司 | 2'-O-(2-methoxyethyl)adenosine and 2'-O-(2-methoxyethyl)guanosine, preparation of derivatives thereof and purifying methods thereof |
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| Title |
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| PROCESS RESEARCH ON THE PREPARATION OF DMT PROTECTED 2-O-METHOXYETHYLGUANOSINE FOR OLIGONUCLEOTIDE SYNTHESIS IN THERAPEUTIC APPLICATIONS;Shabbir Ali S. Taj;《Nucleosides, Nucleotides and Nucleic Acids》;20081231;第27卷;1024-1033页 * |
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