CN116284190A - Pseudo-uridine intermediate, and preparation method and application thereof - Google Patents

Pseudo-uridine intermediate, and preparation method and application thereof Download PDF

Info

Publication number
CN116284190A
CN116284190A CN202211594064.XA CN202211594064A CN116284190A CN 116284190 A CN116284190 A CN 116284190A CN 202211594064 A CN202211594064 A CN 202211594064A CN 116284190 A CN116284190 A CN 116284190A
Authority
CN
China
Prior art keywords
compound
pseudouridine
reaction
solvent
preparation
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202211594064.XA
Other languages
Chinese (zh)
Inventor
李士阔
郝万里
吴梦月
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Anhui University
Original Assignee
Anhui University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Anhui University filed Critical Anhui University
Priority to CN202211594064.XA priority Critical patent/CN116284190A/en
Publication of CN116284190A publication Critical patent/CN116284190A/en
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H23/00—Compounds containing boron, silicon, or a metal, e.g. chelates, vitamin B12
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/04—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H1/00—Processes for the preparation of sugar derivatives
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Saccharide Compounds (AREA)

Abstract

The invention discloses a pseudouridine intermediate, a preparation method and application thereof, and relates to the technical field of drug synthesis. The preparation method comprises the following steps: dissolving a compound 3 in an organic solvent, adding Red-Al under ice bath, and carrying out substitution reaction; after the reaction is finished, adding a diluting solvent for dilution, adding alkali water under ice bath for quenching reaction, filtering and washing with a washing reagent, concentrating the filtrate in vacuum and purifying by a flash chromatography to obtain a compound 5; dissolving the compound 5 prepared in the step (1) in an organic solvent, adding imidazole and TBSCl, stirring for reaction, adding a diluting solvent for dilution after the reaction is finished, extracting by using an extracting solvent, combining organic phases, concentrating the organic phases, and passing through a column to obtain the compound. The beneficial effects are that: the method has the advantages of low cost and low cost of the required reagent; the post-treatment operation of the method is simple; the method has the advantages of short reaction time, easy industrial production and the like.

Description

Pseudo-uridine intermediate, and preparation method and application thereof
Technical Field
The invention belongs to the technical field of medicine synthesis, and particularly relates to a pseudouridine intermediate, a preparation method and application thereof.
Background
Pseudouridine (1) is a metabolite of RNA and has the property of excreting only from the kidneys, and is therefore clinically useful for monitoring the occurrence and development of tumors and diagnosis and treatment of renal diseases. With the deep research on pseudouridine, researchers also find that the level of pseudouridine in urine of patients with gout and psoriasis is obviously increased.
And pseudouridine is an important ring in new crown vaccines of mRNA, which can protect synthetic mRNA from immune system attack, and can also improve molecular stability and protein yield. In this way, researchers add pseudo-uridine and its analogues to mRNA instead of uridine, to solve the problem that mRNA drugs are easily recognized by the immune system and eliminated, resulting in immune side reactions.
One of the key intermediates for pseudouridine synthesis is (R) -2- ((tert-butyldimethylsilyl) oxy) -1- ((4 s, 5R) -5- ((R) - (2, 4-di-tert-butylpyrimidin-5-yl) (hydroxy) methyl) -2, 2-dimethyl-1, 3-dioxolan-4-ethyl) -1-ol (2):
Figure BDA0003996218730000011
compound (2) can be generally synthesized by the following steps, and can undergo 2-step reaction to give pseudouridine (1):
Figure BDA0003996218730000012
wherein, the step 1) is shown that the compound (3) is added with a reducing agent L-selectride and an ether solution of zinc chloride at the temperature of minus 78 ℃ and the raw materials are reacted after the reaction for 10 hours, and then the subsequent treatment process requires the sequential addition of EtOH, H2O, naOH (6 mol/L) and 30% H2O2 so as to achieve the purpose of quenching reaction, the operation is complex, and the product is easy to contain impurities so as to influence the subsequent reaction. In addition, L-selectride is expensive, limiting mass production.
The Chinese patent application document with publication number of CN114702481A discloses a novel synthetic method of pseudouridine, which comprises the following steps: step 1), taking D-ribose as a reaction initiator, and dropwise adding acetyl bromide into an organic solvent to obtain an intermediate I; step 2) taking uracil as a reaction initiator, eliminating active hydrogen by using strong Lewis base in an organic solvent, and adding (Boc) 2O in batches to obtain a corresponding intermediate II; and 3) carrying out condensation reaction on the compound I and the compound II in an organic solvent under the condition of strong Lewis base to obtain an intermediate III, removing a protecting group by using trifluoroacetic acid to obtain an intermediate IV, adding acetic anhydride, carrying out chiral resolution by lipase to obtain a compound V, and finally removing the protecting group under the alkaline condition to obtain a final product VI, namely a pseudo-uridine product. The method adopts mild and safe chemical reagents and enzymatic chiral resolution, shortens the reaction steps, reduces the reaction difficulty, and thus realizes the green synthesis technology bottleneck. However, the method still has the problems of complicated operation steps, high cost, low efficiency and the like, and needs to be further improved.
Disclosure of Invention
The invention aims to solve the technical problems of complex preparation process, complex operation, high cost and low efficiency of the traditional pseudo-uridine intermediate.
The invention solves the technical problems by the following technical means:
a preparation method of a pseudo-uridine intermediate comprises the following steps:
(1) Dissolving a compound 3 in an organic solvent, adding Red-Al under ice bath, and carrying out substitution reaction; after the reaction is finished, adding a diluting solvent for dilution, adding alkali water under ice bath for quenching reaction, filtering and washing with a washing reagent, concentrating the filtrate in vacuum and purifying by a flash chromatography to obtain a compound 5; the structural formula of the compound 3 is
Figure BDA0003996218730000021
The structural formula of the compound 5 is +.>
Figure BDA0003996218730000031
(2) Dissolving the compound 5 obtained in the step (1) in an organic solvent, adding imidazole and TBSCl, stirring, reacting, adding a diluting solvent for dilution after the reaction is finished, extracting with an extracting solvent, combining organic phases, concentrating the organic phases, purifying by a silica gel column to obtain a compound with a structural formula of
Figure BDA0003996218730000032
Compound 2 of (2), i.ePseudouridine intermediates.
The technical route is as follows:
Figure BDA0003996218730000033
the beneficial effects are that: the preparation method for preparing the pseudo-uridine intermediate has the advantages of few steps, simple operation, low cost of required raw material reagents, short reaction time and high efficiency.
Preferably, the molar ratio of compound 3 to Red-Al in step (1) is 1:1.5-2.5.
Preferably, the organic solvent in the step (1) is selected from one or more of dichloromethane, THF, DMF and methanol.
Preferably, the dilution solvent in the step (1) is selected from one or more of dichloromethane, THF, DMF and methanol.
Preferably, the alkali in the step (1) is selected from one of sodium hydroxide and potassium hydroxide.
Preferably, the alkaline water in the step (1) is one of 15% NaOH (aq) and 10% KOH (aq) by mass fraction.
Preferably, the washing reagent in the step (1) is selected from one of dichloromethane, THF, DMF and methanol.
Preferably, the temperature of the substitution reaction in the step (1) is 0-5 ℃ and the time is 1-2h.
Preferably, the molar ratio of compound 5, imidazole and TBSCl in step (2) is 1:2.5-3.5:1.0-1.2.
Preferably, the organic solvent in the step (2) is selected from one or more of THF, DMF, DMAc and methyl tert-butyl ether.
Preferably, the dilution solvent in the step (2) is selected from one of water and ethanol.
Preferably, the extraction solvent in the step (2) is selected from one of ethyl acetate, dichloromethane and methyl tertiary butyl ether.
Preferably, the temperature of the reaction in the step (2) is 20-25 ℃.
The invention also provides a pseudo-uridine intermediate prepared by the method.
The invention also provides an application of the pseudouridine intermediate prepared by the method in preparing the pseudouridine.
The invention has the advantages that:
(1) The preparation method for preparing the pseudo-uridine intermediate has the advantages of few steps, simple operation, low cost of required raw material reagents, short reaction time and high efficiency.
(2) The post-treatment operation of the method is simple.
(3) The method has short reaction time, is easy for industrial production, and can be used for large-scale process production of pseudouridine.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions in the embodiments of the present invention will be clearly and completely described in the following in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1:
the technical route of this embodiment is:
Figure BDA0003996218730000041
a preparation method of a pseudo-uridine intermediate comprises the following steps:
(1) Compound 3 (1.0 g,1.0 eq) was dissolved in 5mLCH 2 Cl 2 (organic solvent), N 2 Three substitutions were made, and a toluene solution (1.1 g) of Red-Al (70 wt.%,2.0 eq) was added under ice bath to carry out substitution reaction at 3℃for 1.5 hours; after the reaction, 20mL of THF (diluting solvent) is added for dilution, the mixture is cooled to 0 ℃ in ice bath, and 15 percent of NaOH aqueous solution is added0.05 mL), and stirring at 0-5 ℃ for 20-30 minutes to quench the reaction; the reaction mixture was filtered through celite and then washed with 30mL of THF (washing reagent), the filtrate was concentrated in vacuo and purified by flash chromatography to give compound 5; compound 3 has the structural formula
Figure BDA0003996218730000051
Compound 5 has the formula +.>
Figure BDA0003996218730000052
(2) After dissolving compound 5 (200 mg,1.0 eq) obtained in step (1) in 5mL of DMF (organic solvent), imidazole (98 mg,3.0 eq) and TBSCl (80 mg,1.1 eq) were added and stirred for 2h at 25℃to allow them to react well, (TLC monitors disappearance of starting material with formation of new spots), after completion of the reaction 5mL of water (dilution solvent) was added for dilution, and the product was extracted with ethyl acetate (10 mL of ethyl acetate solution each for 3 total extractions) followed by washing with water (5 mL of water each for 3 total washes), the organic phases were combined, concentrated at 35-45℃and purified by a silica gel column to give a compound of formula
Figure BDA0003996218730000053
Compound 2 of (a), i.e. pseudouridine intermediate (208 mg, yield 82%).
The calculation process of the yield: 208 (product weight) × 414.50 (raw material molecular weight)/(526.75 (product molecular weight)/(raw material weight) ×100% =82%).
The pseudo uridine intermediate prepared in the present example was subjected to high resolution mass spectrum HRMS detection analysis: M+H+ has the formula C26H29N2O7Si+, calculated 529.3304, measured 529.3315.
Example 2:
the technical route of this embodiment is the same as that of embodiment 1.
A preparation method of a pseudo-uridine intermediate comprises the following steps:
(1) Compound 3 (1.0 g,1.0 eq) was dissolved in 5mLCH 2 Cl 2 (organic solvent), N 2 Three substitutions were made, red-Al (70 wt.%,1.5 eq) toluene solution (1.65 g) and substitution reaction was carried out at 0℃for 2h; after the reaction, 25mL of THF (diluting solvent) is added for dilution, the mixture is cooled to 0 ℃ in ice bath, 10% KOH aqueous solution (0.065 mL) is added, and the mixture is stirred for 20 to 30 minutes at 0 to 5 ℃ for quenching the reaction; the reaction mixture was filtered through celite and then washed with 30mL of methanol (washing reagent), the filtrate was concentrated in vacuo and purified by flash chromatography to give compound 5; compound 3 has the structural formula
Figure BDA0003996218730000061
Compound 5 has the formula +.>
Figure BDA0003996218730000062
(2) After dissolving compound 5 (200 mg,1.0 eq) obtained in step (1) in 5mL of DMF (organic solvent), imidazole (81.7 mg,2.5 eq) and TBSCl (72.7 mg,1.0 eq) were added and stirred for 2h at 22℃to allow them to react well (TLC monitors disappearance of starting material, with formation of new spots), after completion of the reaction, 5mL of water (dilution solvent) was added to dilute the mixture, and the product was extracted with methyl tert-butyl ether (10 mL of ethyl acetate solution each for 3 times), followed by washing with water (5 mL of ethyl acetate solution each for 3 times), the organic phases were combined, concentrated at 35-45℃and purified by a silica gel column to give a compound of the formula
Figure BDA0003996218730000063
Compound 2 of (a), i.e. pseudouridine intermediate (190 mg, yield 75%).
The calculation process of the yield: 190X 414.50/526.75/L. 200×100% =75%.
High resolution mass spectrum HRMS detection analysis was performed on the pseudo uridine intermediate prepared in the present example, and the analysis data were the same as in example 1.
Example 3:
the technical route of this embodiment is the same as that of embodiment 1.
A preparation method of a pseudo-uridine intermediate comprises the following steps:
(1) Compound 3 (1.0 g,1.0 eq) was dissolvedAt 5mLCH 2 Cl 2 (organic solvent), N 2 Three substitutions were made, and a toluene solution (2.75 g) of Red-Al (70 wt.%,2.5 eq) was added under ice bath to carry out substitution reaction at 5℃for 1 hour; after the reaction, 30mL of THF (diluting solvent) is added for dilution, the mixture is cooled to 0 ℃ in ice bath, 15% NaOH aqueous solution (0.12 mL) is added, and the mixture is stirred for 20 to 30 minutes at 0 to 5 ℃ for quenching the reaction; the reaction mixture was filtered through celite and then washed with 30mL of THF (washing reagent), the filtrate was concentrated in vacuo and purified by flash chromatography to give compound 5; compound 3 has the structural formula
Figure BDA0003996218730000071
Compound 5 has the formula +.>
Figure BDA0003996218730000072
(2) After dissolving compound 5 (200 mg,1.0 eq) obtained in step (1) in 5mL of DMF (organic solvent), imidazole (114.3 mg,3.5 eq) and TBSCl (87.3 mg,1.2 eq) were added and stirred at 20℃for a sufficient reaction (TLC monitors disappearance of starting material, with formation of new spots), after completion of the reaction, 5mL of water (dilution solvent) was added for dilution, and the product was extracted with dichloromethane (10 mL of ethyl acetate solution each time for total extraction 3 times), followed by washing with ethanol (5 mL of ethanol each time for total washing with 5mL of water for 3 times), the organic phases were combined, concentrated at 35-45℃and purified by a silica gel column to obtain a compound having the formula
Figure BDA0003996218730000073
Compound 2 of (a), i.e. pseudouridine intermediate (200 mg, yield 79%).
The calculation process of the yield: 200X 414.50/526.75/L200×100% =79%.
High resolution mass spectrum HRMS detection analysis was performed on the pseudo uridine intermediate prepared in the present example, and the analysis data were the same as in example 1.
The above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims (10)

1. A preparation method of a pseudo-uridine intermediate is characterized by comprising the following steps of
(1) Dissolving a compound 3 in an organic solvent, adding Red-Al under ice bath, and carrying out substitution reaction; after the reaction is finished, adding a diluting solvent for dilution, adding alkali water under ice bath for quenching reaction, filtering and washing with a washing reagent, concentrating the filtrate in vacuum and purifying by a flash chromatography to obtain a compound 5; the structural formula of the compound 3 is
Figure FDA0003996218720000011
The structural formula of the compound 5 is +.>
Figure FDA0003996218720000012
(2) Dissolving the compound prepared in the step (1) in an organic solvent, adding imidazole and TBSCl, stirring to react, adding a diluting solvent to dilute after the reaction is finished, extracting with an extracting solvent, combining organic phases, concentrating the organic phases, purifying by a silica gel column to obtain a compound with a structural formula of
Figure FDA0003996218720000013
Compound 2 of (a), i.e. a pseudouridine intermediate.
2. The process for preparing a pseudouridine intermediate according to claim 1, wherein the molar ratio of compound 3 to Red-Al in said step (1) is 1:1.5-2.5.
3. The process for the preparation of a pseudouridine intermediate according to claim 1 or 2, wherein said organic solvent in step (1) is selected from the group consisting of dichloromethane, THF, DMF, methanol in one or more mixtures thereof; the dilution solvent in the step (1) is selected from one or a mixture of more of dichloromethane, THF, DMF and methanol.
4. A process for the preparation of a pseudouridine intermediate according to claim 3, wherein in said step (1) the base is selected from one of sodium hydroxide and potassium hydroxide; the washing reagent in the step (1) is selected from one of dichloromethane, THF, DMF and methanol.
5. The process for producing a pseudouridine intermediate according to claim 4, wherein the temperature of the substitution reaction in said step (1) is 0-5 ℃ for 1-2 hours.
6. The process for preparing a pseudouridine intermediate according to claim 1, wherein said organic solvents in step (2) are each selected from the group consisting of THF, DMF, DMAc, methyl tert-butyl ether and mixtures of one or more thereof; the diluting solvent in the step (2) is selected from one of water and ethanol; the extraction solvent in the step (2) is selected from one of ethyl acetate, dichloromethane and methyl tertiary butyl ether.
7. The process for preparing a pseudouridine intermediate according to claim 1, wherein the molar ratio of compound 5, imidazole and TBSCl in step (2) is 1:2.5-3.5:1.0-1.2.
8. The process for preparing a pseudouridine intermediate according to claim 1, wherein the temperature of the reaction in said step (2) is 20-25 ℃.
9. A pseudouridine intermediate prepared by the method for preparing a pseudouridine intermediate according to any one of claims 1-8.
10. Use of a pseudouridine intermediate prepared by the method for preparing a pseudouridine intermediate according to any one of claims 1-8 for the preparation of pseudouridine.
CN202211594064.XA 2022-12-13 2022-12-13 Pseudo-uridine intermediate, and preparation method and application thereof Pending CN116284190A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202211594064.XA CN116284190A (en) 2022-12-13 2022-12-13 Pseudo-uridine intermediate, and preparation method and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202211594064.XA CN116284190A (en) 2022-12-13 2022-12-13 Pseudo-uridine intermediate, and preparation method and application thereof

Publications (1)

Publication Number Publication Date
CN116284190A true CN116284190A (en) 2023-06-23

Family

ID=86815595

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202211594064.XA Pending CN116284190A (en) 2022-12-13 2022-12-13 Pseudo-uridine intermediate, and preparation method and application thereof

Country Status (1)

Country Link
CN (1) CN116284190A (en)

Similar Documents

Publication Publication Date Title
CN103554201B (en) Gamithromycin preparation method
CN109553610B (en) Preparation method of emtricitabine isomer
CN111646964A (en) Novel method for synthesizing 2H-pyran-2-one derivative by base catalysis
CN116284190A (en) Pseudo-uridine intermediate, and preparation method and application thereof
CN110981851A (en) Preparation method of canagliflozin impurity
CN109824687B (en) Novel synthetic method of xylofuranose derivative
CN111018928B (en) Synthetic method and application of gastrodin hemihydrate
CN112679512B (en) Trabectedin intermediate and preparation method thereof
CN113149973A (en) Synthesis method of engagliflozin
CN114315679A (en) Preparation method of Upactinib chiral intermediate
CN113773281A (en) Preparation method of N-methyl paclitaxel C
CN112778189A (en) (3R,4S) -N-substituent-3-carboxylic acid-4-ethyl pyrrolidine, intermediate and lapatinib
CN115197281B (en) Preparation method of etoposide intermediate
CN106187837B (en) Florfenicol intermediate, preparation method thereof and preparation method of florfenicol
CN104557965A (en) Preparation technology for beta-artemether
CN114957284B (en) Efficient synthesis method and application of natural product Lycibarbitine
CN116178216B (en) Method for synthesizing EP impurity P of aminosalicylic acid
CN114940676B (en) Synthesis method of pseudouridine
CN112679513B (en) Method for preparing key intermediate of koji Bei Ti
CN115322118B (en) Preparation method of benidipine hydrochloride intermediate
CN101805380A (en) Method for preparing (3S, 5S)-2, 3-dihydroxyl-5-hydroxymethyl tetrahydrofuran triacetate
CN108997454B (en) Chemical synthesis method of beta-arbutin
CN114524802A (en) Synthesis method of quinoline compound
CN115703787A (en) Process for preparing 1, 4-disubstituted 2-oxabicyclo [2, 2] octane derivatives
CN116589422A (en) Total synthesis and purification method of high-purity MRI (magnetic resonance imaging) contrast agent intermediate

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination