CN109912672B - Method for glycosylation of base by taking o-alkynyl phenol ether as leaving group - Google Patents

Abstract

The invention discloses a method for constructing a nucleoside C-N glycosidic bond, which has the advantages of simple operation, mild condition and high reaction stereoselectivity and regioselectivity. A glycosylation product shown as a formula P is prepared by taking o-alkynyl phenol ether as a leaving group and carrying out glycosylation reaction on a glycosylation donor shown as a formula I and a glycosylation receptor shown as a formula S under the action of an accelerator in an inert gas environment in the presence of a drying agent, wherein RNH is a base compound, and Gly is glycosyl with one or more hydroxyl groups on a glycosyl ring protected by a protecting group. The glycosylation reaction condition of the invention is mild, the C-N glycosylation product can be obtained with better yield, and the glycosylation method for constructing the nucleoside bond is enriched. The o-alkynyl phenol ether glycosylation donor disclosed by the invention is stable, easy to store and widely used for various glycosylation reactions. The leaving group of the donor is an ether protecting group, and the protecting group operation can be carried out by being distinguished from a benzyl ether protecting group.

Description

Method for glycosylation of base by taking o-alkynyl phenol ether as leaving group

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a method for glycosylation of a basic group by taking an o-alkynyl phenol ether as a leaving group.

Background

The glycosylation reaction is that a glycosyl donor (the anomeric position of the sugar contains a group easy to leave) and an acceptor (a substrate ROH with nucleophilic property, RNH)₂Etc.) are linked together through an acetal bond to form a glycosidic bond. The key and difficult point of glycosylation reaction is the control and yield of the spatial configuration. When the glycosidation reaction is carried out, the acceptor nucleophile can attack the oxygen vat ion formed by the donor from two directions, and then two configurations of alpha and beta are generated. Corresponding measures are generally taken to obtain a product with a specific stereoconfiguration. For example: ortho-base participation, remote participation, conformation of sugar ring, anomeric effect,Additives, solvents, etc. The glycosidation must be carried out strictly anhydrous, otherwise hydrolysis of the donor occurs, reducing the yield of glycosidation. Both donor and acceptor activities, promoters and solvents can affect the yield of glycosidation.

Nucleosides are glycosides of a nitrogenous base condensed with a sugar component, and nucleosides in nucleic acids are formed by condensation of a purine or pyrimidine base with ribose or deoxyribose. The nucleotide as important compound in organism not only is used as the precondition of DNA and RNA, but also is involved in the molecular mechanism of gene information retention, transcription and duplication in organism, and also can be used as the regulator of physiological and biochemical process to be involved in the metabolism of substance in organism. Nucleoside and its derivatives play an extremely important role in the treatment of many important diseases. Development of new drugs for nucleosides (including nucleoside bases) and nucleotides has been the focus of research.

The prior in vitro nucleoside synthesis methods mainly comprise an enzymatic synthesis method and a chemical synthesis method, and the enzymatic synthesis method has the defects of few types of glycosyltransferase and glycosidase, high price and poor universality, so the application of the method is greatly limited. Chemical synthesis methods are currently the main direction of research because they allow the accurate structure to be obtained. The chemical synthesis method of nucleoside compounds is mainly characterized by that it utilizes glycosyl donor and base receptor to form C-N glycosidic bond by means of chemical method. There are many factors that affect the base glycosylation reaction, among which the donor (donor), acceptor (acceptor) and promoter (promoter) are three main factors, and other reaction conditions such as solvent, temperature and concentration have an influence thereon.

The construction of C-N glycosidic linkages of sugars to bases has been a major and difficult point in glycochemistry research. At present, nucleoside drugs are important drugs for clinically treating viral infectious diseases, tumors and AIDS. The medicine saves lives of countless bacteria and fungi infectors in the past decades, and plays a vital role in prolonging the life of human beings. In the chemical synthesis of the drug molecules, the construction of nucleoside bonds is a key step. However, such glycosidic linkages are not particularly easy to construct, mainly for the following reasons:

1) the solubility of the base as an acceptor in this glycosylation is extremely poor, and it cannot be completely dissolved even in DMSO, which makes such glycosylation reaction difficult; 2) since both oxygen and nitrogen atoms have affinity in the pyrimidine-based base, both types of glycosylation products C-O, C-N may be obtained when the glycosylation reaction is carried out. However, there is a problem of regioselectivity between N7 and N9 for purine bases. These problems present a great challenge to the precise construction of base glycosidic linkages.

Fisher and Helferich, two scientists, to solve the problem of insolubility of bases, first proposed the synthesis of nucleosides using base metal salts.^[1]They used halogenated sugar and silver salt of basic group in xylene as solvent, and take nucleoside product under heating condition. Unfortunately, the yield is too low, and some compounds are basically not reacted, and the yield is improved by later changing the basic silver salt into basic mercury salt, but the mercury is a toxic substance of heavy metal, so that the method is extremely destructive to the environment, and the method is limited to be used.

In 1930, Hilbert and Johnson reacted fully acetylated glycochlorin with 2, 4-diethoxypyrimidine, and the pyrimidine glucoside was successfully synthesized by acidification.^[2]The method can obtain the glycosylation product through one-step acidification under the heating condition, is convenient and quick to operate, does not need the participation of heavy metal, but has the defects that the glycosylation yield is not high, and the byproducts are more, such as: nitrogen alkylated products and O-glycosidated products, among other things, have the additional disadvantage that the product also presents some difficulty in dealkylating the nucleated glycoside.

In 1960, the Sato group combined the total acetyl protected ribose with an equimolar amount of N⁶Acetyl adenine is used as an accelerant in the condition of high-temperature melt condensation to obtain corresponding acetyl protected adenosine,^[3]the method has no universality (only limited to a substrate with a low melting point), and simultaneously, the stereoselectivity and the regioselectivity of the reaction are poor, so the method is not widely applied to the synthesis of nucleosides.

In 1963, Birkofer, Nishimura, wittenburg et al, performed some modifications to improve the solubility of bases in organic solvents based on the classical Hilbert-Johnson reactionThe bases are first silanized. Solves the problem of poor solubility and low activity of basic groups in organic solvents, thereby improving the efficiency of C-N bond synthesis. [⁴]And the silylation of the basic group is greatly improved in nucleophilicity compared with the alkylation, and the deprotection of the silicon group is relatively easy. The disadvantage is that it requires the use of equivalent mercury salt or silver perchlorate as an accelerator, which is very environmentally hazardous and very explosive at high temperatures, and therefore this method is not the best way to synthesize a glycosidic bond.

In 1970, Vorbruggen first reacted an acylsugar with a silanized base in SnCl based on the Silyl-Hibert-Johnson method₄The nucleoside is synthesized under the action, and a very good effect is achieved.^[5]Compared to the Silyl-Hilbert-Johnson reaction, the Vorbruggen reaction uses peracetylated sugars as donors, the donors of this method are more stable and easier to prepare, and the Vorbruggen reaction uses SnCl₄、TiCl₄、FeCl₃、BF₃.Et₂Lewis acids such as O, TMSOTf and the like are used as accelerators, so that the pollution to the environment is reduced. The method is a mode for preparing nucleoside which is most widely applied at present, but has the disadvantages of harsh conditions, requirement of using equivalent or more activating agents, poor reaction effect of partial substrates and unsatisfactory yield. The regioselectivity of the glycosylation product is also not very good when the acceptor is a purine base, and is generally a mixture of N7/N9.

In 1991, the Knapp group utilized the reaction of glycosyl donors of thiophenols with purines and pyrimidines activated with trimethylsilyl,^[6]in 1992, the project group of Jean. Marie Beau was synthesized using sulfoxide glycosides with equivalent amounts of TMSOTf and silyl activated base pair nucleosides,^[7]the C-N bond of nucleoside bond and other sugar is constructed in good yield, and an effective method is provided for the synthesis of nucleoside. Nevertheless, this method is unsatisfactory in that the nucleoside synthesis of thioglycosides requires equivalent amounts of NIS, TsOH as activator and the nucleoside synthesis of sulfoxide glycosides requires equivalent amounts of TMSOTf as activator, both of which are too acidic to be suitable for acid sensitive substrates.

In 1993, the group of Lawrence J.Marnet topics produced nucleoside products in moderate yields but with poor selectivity using a pentenyl glycosyl donor reacted with purines using NIS, TsOH or neutral iodonium perchlorate (IDCP) as activator^[8]。

The method for constructing the C-N glycosidic bond of the nucleoside based on the importance of the construction of the C-N bond of the nucleoside in the research of novel nucleoside and nucleotide drugs has the advantages of simple development operation, mild condition, simple hotspot operation, mild condition and high stereoselectivity and regioselectivity of reaction.

Reference documents:

[1]Fischer,E.；Helferich,B.Chem.Ber.1974,47,210.

[2]Hilbert,G.E.；Johnson,T.B.J.Am.Chem.Soc.1930,52,4489.

[3]Diekmann,E.；Friedrich,Jr.；Fritz,H-G.J.Prakt.Chem.1993,335,415.

[4]Birkofer,L.；Ritter,A.；Kiihlthau,H.P.Angew.Chem.1963,4,209.

[5]Niedball,U.；Vorbrugg,H.Angew.Chem.Int.Ed.1970,9,461.

[6]Knapp,S.；Shieh,W.-C.Tetrahedron Lett.1991,32,3627-3630.

[7]Chanteloup,L.；Beau,J.-M.Tetrahedron Lett.1992,33,5347-5350.

[8]Chapeau,M.-C.；Marnett,L.J.Org.Chem.1993,58,7258.

disclosure of Invention

The technical problem to be solved by the invention is to provide a method for constructing a C-N glycosidic bond of a nucleoside, which has the advantages of simple operation, mild condition, high reaction stereoselectivity and regioselectivity.

In order to solve the technical problems, the invention adopts the technical scheme that: a method for glycosylation of a base with an o-alkynyl phenol ether as a leaving group.

In an inert gas environment, in the presence of a drying agent and under the action of an accelerating agent, a glycosylation donor shown as a formula I and a glycosylation receptor shown as a formula S are subjected to glycosylation reaction as shown in the specification to obtain a glycosylation product shown as a formula P,

wherein RNH is a basic group compound, Gly is glycosyl with one or more hydroxyl groups on a glycosyl ring protected by a protecting group, and the temperature of glycosylation reaction is-50 to-20 ℃; preferably-40 to-35 ℃;

the base is a pyrimidine or a purine,

when the base is a pyrimidine, the promoters are BSTFA, NIS and TMSOTf;

when the base is a purine, the enhancers are NIS and TMSOTf.

The base-based compound RNH is selected from any one of compounds represented by the following formulas S1 to S3,

wherein R' is an acyl protecting group.

The acyl protecting group is selected from Bz, Cbz or Boc, and preferably Boc.

The glycosylation donor is selected from any one of the compounds shown below,

。

when the base is purine, the molar ratio of RNH to TMSOTf to NIS is 1:0.1: 1-1: 1:2, preferably 1:0.6: 1.5-1: 1: 2; the molar ratio of the glycosylation donor to the RNH is 1: 1-5: 1, preferably 1.2: 1-2: 1; the mass-volume ratio of the glycosylation donor to dichloromethane is 20-100 mg/mL, and the glycosylation reaction time is 3-15 hours.

When the base is pyrimidine, the molar ratio of RNH to TMSOTf, NIS to BSTFA is 1:0.1:1: 1-1: 1:2:4, preferably 1:0.6:1: 1-1: 1:2:4, and the molar ratio of the glycosylation donor to RNH is 1: 1-5: 1, preferably 1:0.6: 1.5-1: 1: 2; the mass-volume ratio of the glycosylation donor to dichloromethane is 20-100 mg/mL, and the glycosylation reaction time is 3-15 hours.

Preferably, the inert gas is selected from argon or nitrogen; preferably, the drying agent is selected from acid-washed molecular sieves. In the present invention, the product compound P comprises the following molecules:

in the invention, C-N glycosylation products can be respectively obtained with yield of 60-100% in glycosylation reactions of the o-alkynyl phenol ether donor and the amino sugar receptor, the reaction effect of pyrimidine bases in the glycosylation reactions is obviously better than that of purine bases, mainly because the affinity of purine bases is weaker, some reactions are incomplete in the reaction process, and the yield is lower.

Has the advantages that: the glycosylation reaction condition of the invention is mild, the C-N glycosylation product can be obtained with better yield, and the glycosylation method for constructing the nucleoside bond is enriched. The o-alkynyl phenol ether glycosylation donor disclosed by the invention is stable, easy to store and widely used for various glycosylation reactions. The leaving group of the donor is an ether protecting group, and the protecting group operation can be carried out by being distinguished from a benzyl ether protecting group.

Detailed Description

The invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the description of the embodiments is only for illustrating the present invention and should not be taken as limiting the invention as detailed in the claims.

In the present invention, abbreviations have the following meanings:

BSTFA: n, O-bis (trimethylsilyl) trifluoroacetamide

NIS: n-iodosuccinimide

TMSOTf: trimethylsilyl trifluoromethanesulfonate

Boc: tert-butyloxycarbonyl radical

Bz: benzoyl radical

Cbz: benzyloxycarbonyl group

Preparation of a compound of formula I:

1. compound I1^[9]The preparation of (1):

solid compound 1(5.0g,9.1mmol), Ph₃P(471.7mg,3.6mmol)，Pd(PPh₃)₂Cl₂(637.7mg,1.0mmol), CuI (692.1mg,3.6mmol) methoxyphenylacetylene (1.8g,13.6mmol) was placed in a nitrogen-blanketed round-bottomed flask, cooled to-78 ℃ and then purged with a vacuum diaphragm pump. DMF (10mL) was then added for purging and iPr was added₂The reaction was carried out by purging with NH (20mL) three times, then the reaction mixture was allowed to stand at room temperature for 15min, and then the reaction mixture was allowed to stand in an oil bath at 70 ℃ for 4 hours. Point plate monitoring for complete addition of NH to be reacted₄Cl extraction, dilution with ethyl acetate, filtering with celite and silica gel, and filtering the filtrate with saturated NH₄Three times with Cl and two times with saturated NaCl to combine the organic phases. With anhydrous Na₂SO₄And (5) drying. And (4) padding with diatomite and silica gel for filtration, and performing spin drying on the filtrate and then performing column packing. Chromatography with (PE/EA ═ 4:1) afforded I1(4.4g, 87.1%) as a pale yellow solid.

Reference documents: [9] hu, y.; yu, k.; shi, l.; liu, l.; sui, j.; sun, J.J.Am.chem.Soc.2017,139,12736-12744.

2. Compound I2^[9]The preparation of (1):

compound I1(3.0g,5.4mmol) was dissolved in CH3OH (10.0mL) and saturated CH was added at room temperature₃Na solution (0.5ml), after reacting for 2 hours, was checked by TLC for completion, neutralized to pH 7 with acidic resin, filtered with suction, spun-dried, and then dissolved in Py (10.0ml) in an ice-water bathBenzoyl chloride (3.3ml,27.0mmol) was added dropwise, the reaction was allowed to proceed at room temperature for 3 hours, and the reaction was monitored by spotting plates for the completion of the reaction by the addition of CH₃OH extraction and inactivation reaction, diluting with a large amount of ethyl acetate, washing with saturated NaCl twice, washing with 1NHCl twice, washing with saturated NaCl twice, mixing organic phases, and adding anhydrous Na₂SO₄Drying, suction filtration, spin drying of the filtrate and recrystallization gave I2 as a white solid (3.9g, 90.5%).

Reference documents: [9] hu, y.; yu, k.; shi, l.; liu, l.; sui, j.; sun, J.J.Am.chem.Soc.2017,139,12736-12744.

3. Preparation of Compound I3

Experimental work on the synthesis referring to the synthetic procedure of I1, p-methoxyphenylacetylene (876.5mg,6.6mmol) and 2(3.0g,4.4mmol) reacted completely to give the pale yellow compound I3(4.4mg, 87.1%). [ alpha ] to]²⁸ _D＝-74.4°(c＝1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ8.01–7.94(m,2H),7.85–7.77(m,2H),7.61–7.54(m,1H),7.51–7.40(m,8H),7.40–7.26(m,7H),7.08–7.01(m,1H),7.00–6.92(m,2H),6.02–5.84(m,3H),5.77(s,1H),4.50(dd,J＝10.3,4.9Hz,1H),4.31(t,J＝9.5Hz,1H),4.21(td,J＝9.7,4.9Hz,1H),4.00(t,J＝10.1Hz,1H),3.85(s,3H)；¹³C NMR(100MHz,Acetone-d₆)δ206.2,166.0,165.7,160.7,157.8,138.5,134.2,134.1,133.9,133.8,130.4,130.3,130.2,130.2,129.7,129.4,129.2,128.8,127.1,123.4,116.2,115.5,114.7,114.5,99.6,94.6,84.1,79.1,73.2,73.1,69.0,67.4,55.7,30.4,30.2,30.0,29.8,29.6,29.4,29.2；HRMS(ESI)calcd for C₄₂H₃₄O₉Na[M+Na]⁺705.2095,found 705.2080.

Preparation of Compound I4

Experimental procedure for synthesis of I4 reference is made to the synthesis procedure of I1:reaction of p-methoxyphenylacetylene (1.8g,13.6mmol) and 3(5.0g,9.1mmol) was complete to give the pale yellow compound I4(4.5g, 89.6%). [ alpha ] to]_D ²⁵＝-66.3(c 1.0,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ7.59–7.53(m,2H),7.48(dd,J＝7.7,1.7Hz,1H),7.28–7.22(m,1H),7.13–6.99(m,2H),6.92–6.83(m,2H),5.66(dd,J＝10.5,8.0Hz,1H),5.47(dd,J＝3.4,1.0Hz,1H),5.17–5.07(m,2H),4.25(dd,J＝11.2,6.9Hz,1H),4.16(dd,J＝11.2,6.2Hz,1H),4.12–4.07(m,1H),3.81(s,3H),2.18(s,3H),2.06(s,3H),2.00(s,3H),1.90(s,3H)；¹³C NMR(100MHz,CDCl₃)δ170.5,170.4,170.3,169.6,159.7,157.1,133.5,133.4,129.2,123.1,115.6,115.5,114.5,113.9,100.1,94.2,83.4,77.5,77.1,76.8,71.1,70.9,68.3,67.0,61.5,55.3,20.8,20.7,20.7；HRMS(ESI)calcd for C₂₉H₃₀O₁₁Na[M+Na]⁺577.1680,found 577.1656.

5. Preparation of Compound I5

Experimental work for Synthesis of I5 referring to the synthetic procedure for I1, I4(3.0g,5.4mmol) was deacetylated and reacted with BzCl (3.1ml,27.1mmol) to give I5(4.0g, 93.0%) as a white solid. [ alpha ] to]_D ²⁵＝+137.7(c1.0,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ8.20–8.13(m,2H),8.08(dd,J＝8.2,1.4Hz,2H),7.87–7.81(m,2H),7.73(dd,J＝8.1,1.4Hz,2H),7.67–7.58(m,2H),7.55–7.41(m,6H),7.40–7.31(m,3H),7.25(qd,J＝8.7,7.8,1.4Hz,3H),7.17(t,J＝7.7Hz,2H),7.10(td,J＝7.9,1.8Hz,1H),7.01(td,J＝7.5,1.1Hz,1H),6.86–6.78(m,2H),6.25(dd,J＝10.4,8.0Hz,1H),6.10(d,J＝3.5Hz,1H),5.71(dd,J＝10.4,3.5Hz,1H),5.50(d,J＝8.0Hz,1H),4.70(dt,J＝10.4,5.2Hz,1H),4.63–4.51(m,2H),3.83(s,3H)；¹³C NMR(100MHz,CDCl₃)δ166.12,165.7,165.7,165.3,159.5,157.2,133.8,133.5,133.5,133.5,133.3,133.0,130.2,129.9,129.9,129.8,129.5,129.2,129.1,128.9,128.8,128.8,128.6,128.4,128.1,123.0,115.6,115.5,114.6,113.7,100.3,94.2,83.2,77.5,77.1,76.8,72.0,71.9,69.2,68.1,62.4,55.4；HRMS(ESI)calcd for C₄₉H₃₈O₁₁Na[M+Na]⁺825.2306,found 825.2270.

6. Preparation of Compound I6

Experimental procedure for the Synthesis of Compound I6 referring to the synthetic procedure of I1, p-methoxyphenylacetylene (985.1mg,6.7mmol) and 5(3.0g,4.5mmol) were reacted to give compound I6(2.7mg, 90.0%) as pale yellow. [ alpha ] to]²⁸ _D＝+29.6(c1.0,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ8.02(d,J＝7.3Hz,2H),7.95(d,J＝7.6Hz,2H),7.90(d,J＝7.7Hz,2H),7.60–7.49(m,4H),7.42(q,J＝8.2,7.8Hz,4H),7.35–7.26(m,4H),7.20(dd,J＝16.3,7.4Hz,2H),6.99(td,J＝7.4,1.3Hz,1H),6.89–6.78(m,2H),6.15(dd,J＝7.2,4.7Hz,1H),6.08(d,J＝4.7Hz,1H),6.02(s,1H),4.86(dt,J＝7.2,4.6Hz,1H),4.75(dd,J＝11.9,4.3Hz,1H),4.59(dd,J＝12.0,4.8Hz,1H),3.77(s,3H)；¹³C NMR(100MHz,CDCl₃)δ166.3,165.4,165.2,159.6,156.3,133.7,133.5,133.2,133.1,133.0,129.9,129.8,129.8,129.5,129.2,129.1,128.9,128.6,128.4,128.3,122.8,115.9,115.5,114.8,114.1,104.1,94.4,84.0,79.6,75.8,72.2,65.9,64.3,55.3,15.4；HRMS(ESI)calcd for C₄₁H₃₂O₉Na[M+Na]⁺691.1939,found 691.1912.

7. Preparation of Compound I7

Experimental procedure for the Synthesis of Compound I7 with reference to the synthetic procedure of I1, p-methoxyphenylacetylene (1.0g,7.8mmol) and 6(3.5mg,5.2mmol) were reacted to give the pale yellow compound I7(2.9g, 83.7%). [ alpha ] to]²⁸ _D＝-72.4(c 1.0,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ8.08(ddd,J＝8.4,3.2,1.4Hz,4H),7.95–7.87(m,2H),7.59(tt,J＝7.2,1.3Hz,1H),7.53–7.38(m,5H),7.38–7.27(m,8H),7.08(td,J＝7.4,1.4Hz,1H),6.84–6.77(m,2H),5.89(t,J＝6.1Hz,1H),5.77(dd,J＝6.2,4.4Hz,1H),5.71(d,J＝4.4Hz,1H),5.42(td,J＝5.7,3.6Hz,1H),4.78(dd,J＝12.5,3.7Hz,1H),3.98(dd,J＝12.5,5.6Hz,1H),3.83(s,3H)；¹³C NMR(100MHz,CDCl₃)δ160.4,160.2,160.0,154.3,151.9,128.3,128.3,128.2,128.0,127.8,124.8,124.8,124.7,124.1,124.1,123.9,123.7,123.3,123.2,123.0,117.9,111.3,110.2,109.8,108.6,93.7,88.9,78.6,72.2,72.1,71.9,71.6,64.4,64.1,63.4,55.9,50.1；HRMS(ESI)calcd for C₄₁H₃₂O₉Na[M+Na]⁺691.1939,found691.1908.

8. Preparation of Compound I8

Experimental work on the Synthesis of Compound I8 referring to the synthetic procedure of I1, p-methoxyphenylacetylene (436.1mg,3.3mmol) and 7(1.5g,2.2mmol) were reacted to completion to give the pale yellow compound I8(1.2g, 83.2%). [ alpha ] to]²⁸ _D＝111.7(c 1.0,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ8.08–7.97(m,4H),7.94–7.85(m,2H),7.61–7.54(m,1H),7.50–7.39(m,5H),7.33–7.26(m,8H),7.24(dd,J＝8.4,1.3Hz,1H),7.06(td,J＝7.4,1.3Hz,1H),6.78–6.69(m,2H),6.06(dd,J＝7.5,5.4Hz,1H),5.82–5.74(m,2H),5.54(d,J＝5.5Hz,1H),4.62(dd,J＝12.5,5.1Hz,1H),4.07(dd,J＝12.4,2.6Hz,1H),3.78(s,3H)；¹³C NMR(100MHz,CDCl₃)δ165.6,165.2,159.4,157.2,133.5,133.4,133.3,133.2,133.0,123.0,129.9,129.8,129.3,129.3,129.0,128.9,128.5,128.3,128.2,123.1,116.6,115.4,115.0,113.7,99.6,93.9,83.8,77.3,77.2,77.0,76.7,69.9,69.6,67.7,61.9,55.2；HRMS(ESI)calcd for C₄₁H₃₂O₉Na[M+Na]⁺691.1939,found 691.1940.

Preparation of a compound of formula S:

1. preparation of Compound S1^[10]

Cytosine Sa1(500mg,2.7mmol), DMAP (33.0g,0.27mmol) was dissolved in dry THF under nitrogen, and Boc was added slowly in an ice-water bath₂O (3.9g,10.8mmol) and reacted at room temperature overnight, the system became a clear orange solution which was quenched with methanol after TLC detection of the completion of the reaction. Then, the mixture was diluted with a large amount of EA, washed with a saturated aqueous sodium hydrogencarbonate solution and saturated NaCl, dried over anhydrous sodium sulfate, and concentrated. After two hours on the column, flash column through (PE/EA ═ 1:1) gave product S1 as a white solid (1.1g, 82.0%).

2. Preparation of Compound S2^[10]

Adenine Sa2(300mg,2.0mmol), DMAP (48.5g,0.04mmol) was dissolved in dry THF under nitrogen, and Boc was added slowly in an ice-water bath₂O (2.6g,11.9mmol), at room temperature overnight, until the system became an orange clear solution, the reaction was completely quenched with methanol by TLC. Then diluted with a large amount of EA, separately with saturated NaHCO₃Washing with aqueous solution and saturated NaCl, anhydrous Na₂SO₄Drying and concentrating. Flash column chromatography (PE/EA ═ 5:1) afforded product Sb2(786.1mg, 91.0%) as a white solid.

Sb2(500mg,1.1mmol) was dissolved in methanol (13ml) and then saturated NaHCO (6.5ml) was added₃The solution is put in an oil bath at 50 ℃ for reaction for 0.5h, diluted by a large amount of EA after TLC detection reaction is completed and washed by water for 2 times, and then washed by saturated NaCl for 2 times, and anhydrous Na₂SO₄Dry, concentrate, go to column and flash column through (PE/EA ═ 1:1) to give product S2(306.1mg, 79.5%) as a white solid.

3. Preparation of Compound S3^[10]

Experimental procedure for synthesis of S3 reference is made to the synthesis procedure of S2: compound Sa3(300mg,1.8mmol), DMAP (24.4mg,0.2mmol), Boc₂O (2.6g,11.9mmol) gave compound S3(510.9mg, 78.1%).

[10]Andrea,P.；Giampaolo,G.；Ivana,P.；Mariolino,C.；Giammario,N.Eur.J.Org.Chem.2008,34,5786–5797.

Example 1:

compound S1(20mg,0.064mmol) was dissolved in dry DCM (1mL) under nitrogen, BSTFA (68.3. mu.L, 0.26mmol) was slowly added dropwise at room temperature for 0.5h, then NIS (29mg,0.13mmol) and I1(53.2mg,0.096mmol) were rapidly added thereto, then placed in a low temperature reactor at-35 ℃ and TMSOTf (7.0. mu.L, 0.038mmol) was slowly added after 10min of reaction, at which temperature reaction overnight was detected by a TCL plate and the reaction was complete and detected by Et₃N quench and spin dry on column (PE/EA ═ 3:1) to give compound P1(40.0mg, 96.8%). [ alpha ] to]_D ²⁵＝+18.0(c0.5,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ7.62(d,J＝7.7Hz,1H),7.15(d,J＝7.7Hz,1H),6.09(d,J＝9.5Hz,1H),5.39(t,J＝9.4Hz,1H),5.14(dt,J＝19.5,9.6Hz,2H),4.22(dd,J＝12.5,5.1Hz,1H),4.07(dd,J＝12.4,1.8Hz,1H),3.97–3.88(m,1H),2.05(s,3H),2.03(s,3H),1.98(s,3H),1.94(s,3H),1.52(s,18H)；¹³C NMR(100MHz,CDCl₃)δ170.6,169.8,169.6,162.7,154.1,149.3,143.2,97.3,85.3,81.1,77.5,77.1,76.8,75.1,72.8,70.0,68.0,61.8,27.7,20.8,20.6,20.6,20.4；HRMS(ESI)calcd for C₂₈H₃₉N₃O₁₄K[M+K]⁺680.20636,found 680.2032.

Example 2:

experimental work for synthesis of P2 reference is made to the synthesis procedure of P1: s1(20mg,0.064mmol) was reacted with BSTFA (68.3. mu.L, 0.26mmol) under nitrogen for 2.5h, followed by addition of I2(77.1mg,0.096mmol), NIS (29mg,0.13mmol) and TMSOTf (7.0. mu.L, 0.038mmol) overnight to afford compound P2(57.3mg, 100%). [ alpha ] to]_D ²⁵＝+17.8(c 0.5,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ8.49(d,J＝7.6Hz,1H),8.10–8.04(m,2H),7.98–7.93(m,2H),7.89–7.83(m,2H),7.84–7.78(m,2H),7.65–7.33(m,13H),7.04(d,J＝7.7Hz,1H),6.65(d,J＝9.3Hz,1H),6.29(t,J＝9.5Hz,1H),6.01(dt,J＝16.4,9.6Hz,2H),4.88(ddd,J＝10.0,4.7,2.8Hz,1H),4.72(dd,J＝12.5,2.8Hz,1H),4.64(dd,J＝12.5,4.7Hz,1H),1.48(s,18H)；¹³C NMR(100MHz,Acetone-d₆)δ166.4,166.1,165.9,165.8,163.4,154.2,150.2,146.2,134.6,134.5,134.4,134.1,130.8,130.5,130.5,130.2,130.0,130.0,129.6,129.5,129.4,129.4,97.4,85.4,82.1,75.6,74.4,72.6,69.8,63.6,27.8；HRMS(ESI)calcd for C₄₈H₄₇N₃O₁₄Na[M+Na]⁺912.2950,found 912.2947.

Example 3:

experimental procedure for the synthesis of P3 reference was made to the synthetic procedure for P1 to yield compound P3(43.1mg, 87.1%). [ alpha ] to]_D ²⁵＝+21.7(c1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ8.38(d,J＝7.7Hz,1H),7.96–7.91(m,2H),7.88–7.82(m,2H),7.59–7.53(m,2H),7.46–7.38(m,6H),7.35–7.30(m,3H),7.07(d,J＝7.7Hz,1H),6.55(d,J＝9.3Hz,1H),6.12(t,J＝9.4Hz,1H),5.88(t,J＝9.3Hz,1H),5.74(s,1H),4.46(dd,J＝10.3,4.9Hz,1H),4.36(t,J＝9.5Hz,1H),4.27(td,J＝9.6,4.8Hz,1H),3.97(t,J＝10.1Hz,1H),1.48(s,18H)；¹³C NMR(100MHz,Acetone-d₆)δ165.1,165.0,162.5,153.2,149.2,145.3,137.5,133.6,133.4,129.6,129.5,129.4,128.8,128.7,128.5,128.5,128.0,126.2,101.4,96.4,84.5,81.9,78.1,72.5,72.2,69.2,67.9,29.6,29.5,29.4,29.2,29.0,28.8,28.6,28.4,26.9；HRMS(ESI)calcd for C₄₁H₄₃N₃O₁₂Na[M+Na]⁺792.2739,found 792.2697.

Example 4:

experimental procedure for the synthesis of P4 reference was made to the synthetic procedure for P1 to yield compound P4(41.2mg, 100%). [ alpha ] to]_D ²⁵＝+29.4(c 1.0,CHCl₃)；¹H NMR(400MHz,Chloroform-d)δ7.65(d,J＝7.7Hz,1H),7.14(d,J＝7.7Hz,1H),6.04(d,J＝9.0Hz,1H),5.46(d,J＝3.1Hz,1H),5.31–5.24(m,1H),5.21(dd,J＝10.2,3.2Hz,1H),4.16–4.00(m,3H),2.15(s,3H),2.00(s,3H),1.94(d,J＝4.3Hz,6H),1.51(s,18H)；¹³C NMR(100MHz,CDCl₃)δ170.4,170.0,169.9,169.7,162.6,154.1,149.2,143.6,97.2,85.2,81.4,77.5,77.2,76.8,73.9,70.9,67.8,67.2,67.1,61.4,27.7,20.7,20.7,20.5；HRMS(ESI)calcd for C₂₈H₃₉N₃O₁₄K[M+K]⁺680.20636,found 680.2032.

Example 5:

experimental procedure for the synthesis of P5 reference was made to the synthetic procedure for P1 to yield compound P5(53.3mg, 93.2%). [ alpha ] to]_D ²⁵＝+70.6(c 1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ8.43(d,J＝7.7Hz,1H),8.25–8.20(m,2H),8.03–7.98(m,2H),7.90–7.86(m,2H),7.81–7.76(m,2H),7.76–7.69(m,1H),7.65–7.54(m,4H),7.53–7.45(m,3H),7.39(t,J＝7.8Hz,2H),7.35–7.29(m,2H),7.14(d,J＝7.6Hz,1H),6.69(d,J＝9.2Hz,1H),6.26(dd,J＝3.3,1.2Hz,1H),6.19(dd,J＝10.0,3.4Hz,1H),6.08(t,J＝9.5Hz,1H),5.14(td,J＝6.3,1.2Hz,1H),4.70(dd,J＝11.4,6.6Hz,1H),4.61(dd,J＝11.5,6.0Hz,1H),1.48(s,18H)；¹³C NMR(100MHz,Acetone-d₆)δ166.2,166.2,165.9,165.6,163.4,154.2,150.1,145.9,134.6,134.5,134.3,134.1,130.7,130.5,130.4,130.3,130.2,130.1,129.9,129.7,129.5,129.4,129.3,97.6,85.3,82.4,74.9,72.8,70.5,69.6,63.0,30.4,30.2,30.0,29.8,29.6,29.4,29.2,27.7；HRMS(ESI)calcd for C₄₈H₄₇N₃O₁₄K[M+K]⁺928.26896,found 928.2697.

Example 6:

experimental procedure for the synthesis of P6 reference was made to the synthetic procedure for P1 to yield compound P6(48.5mg, 90.1%). [ alpha ] to]_D ²⁵＝-12.8(c 1.0,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ8.11–8.06(m,2H),7.93(ddd,J＝11.1,8.3,1.4Hz,4H),7.81(d,J＝7.6Hz,1H),7.61–7.49(m,3H),7.46(t,J＝7.8Hz,2H),7.35(td,J＝7.7,5.1Hz,4H),7.03(d,J＝7.6Hz,1H),6.26(d,J＝3.9Hz,1H),5.92(t,J＝5.9Hz,1H),5.85(dd,J＝5.8,3.9Hz,1H),4.83(dd,J＝12.0,2.9Hz,1H),4.76(dt,J＝6.4,3.5Hz,1H),4.70(dd,J＝12.0,4.3Hz,1H),1.55(s,18H)；¹³C NMR(100MHz,CDCl₃)δ166.2,165.3,165.3,162.9,153.9,149.4,143.8,133.7,133.7,133.6,130.0,129.9,129.7,129.4,128.7,128.5,96.8,90.9,85.2,80.5,77.5,77.2,76.8,74.9,71.0,63.8,27.8；HRMS(ESI)calcd for C₄₀H₄₁N₃O₁₂K[M+K]⁺794.23218,found 794.2314.

Example 7:

experimental procedure for the synthesis of P7 reference was made to the synthetic procedure for P1 to yield compound P7(48.5mg, 100.0%). [ alpha ] to]_D ²⁵＝+27.0(c 0.25,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ7.99–7.93(m,2H),7.89–7.84(m,4H),7.81(d,J＝7.7Hz,1H),7.58–7.52(m,1H),7.50–7.43(m,2H),7.41(t,J＝7.7Hz,2H),7.32(q,J＝7.8Hz,4H),7.18(d,J＝7.6Hz,1H),6.36(d,J＝9.5Hz,1H),6.07(t,J＝9.6Hz,1H),5.64(t,J＝9.5Hz,1H),5.48(td,J＝10.1,5.6Hz,1H),4.51(dd,J＝11.5,5.7Hz,1H),3.80(t,J＝11.0Hz,1H),1.49(s,18H)；¹³C NMR(100MHz,CDCl₃)δ165.7,165.5,165.5,162.7,154.2,149.3,143.3,133.8,133.5,130.2,130.0,129.9,128.8,128.7,128.7,128.6,128.5,128.1,97.4,85.2,82.1,77.5,77.2,76.8,72.8,70.7,69.8,66.1,27.7；HRMS(ESI)calcd for C₄₀H₄₁N₃O₁₂K[M+K]⁺794.23218,found 794.2306.

Example 8:

experimental procedure for the synthesis of P8 reference was made to the synthetic procedure for P1 to yield compound P8(46.6mg, 96.0%). [ alpha ] to]_D ²⁵＝+132.7(c 1.0,CHCl₃)；¹H NMR(400MHz,CDCl₃)δ8.14–8.07(m,2H),7.93–7.81(m,5H),7.68–7.60(m,1H),7.53(t,J＝7.6Hz,2H),7.50–7.41(m,2H),7.30(dt,J＝20.3,7.7Hz,4H),7.19(d,J＝7.6Hz,1H),6.38(d,J＝9.4Hz,1H),5.98(t,J＝9.7Hz,1H),5.82(d,J＝3.5Hz,1H),5.77(dd,J＝9.9,3.4Hz,1H),4.41(dd,J＝13.5,1.9Hz,1H),4.16(d,J＝13.4Hz,1H),1.49(s,18H)；¹³C NMR(100MHz,CDCl₃)δ165.7,165.5,165.3,162.6,154.1,149.2,143.3,133.7,133.7,133.5,130.07,129.8,129.8,129.4,128.8,128.7,128.5,128.4,128.3,97.4,85.1,82.1,77.5,77.1,76.8,71.9,69.3,68.8,67.6,27.7；HRMS(ESI)calcd for C₄₀H₄₁N₃O₁₂K[M+K]⁺794.2322,found 794.2305.

Example 9:

under nitrogen protection, compounds S2(20mg,0.060mmol) and I3(48.8mg,0.072mmol) were dissolved in dry DCM, stirred at room temperature for 0.5h, NIS (20.1mg,0.09mmol) was added rapidly thereto, and the mixture was subjected to a low temperature reaction at-350 deg.CAfter 10min, TMSOTf (6.5. mu.L, 0.036mmol) was slowly added to the reaction vessel and allowed to react at this temperature overnight, and Et was detected on a TCL plate after completion of the reaction₃N quench and spin dry on column (PE/EA ═ 4:1) to give compound P10(37.9mg, 80.0%). [ alpha ] to]_D ²⁵＝+23.5(c 1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ8.90(s,1H),8.82(s,1H),7.98–7.93(m,2H),7.72–7.67(m,2H),7.60–7.54(m,1H),7.52–7.41(m,5H),7.36–7.28(m,5H),6.65(d,J＝9.2Hz,1H),6.41(t,J＝9.2Hz,1H),6.21(t,J＝9.4Hz,1H),5.82(s,1H),4.50–4.43(m,2H),4.38(td,J＝9.6,4.8Hz,1H),4.02(t,J＝10.0Hz,1H),1.30(s,18H)；¹³C NMR(100MHz,Acetone-d₆)δ166.0,165.4,154.0,153.0,151.3,150.7,145.0,138.4,134.6,134.4,130.3,130.3,129.8,129.5,129.3,129.1,129.1,128.9,127.1,102.3,83.8,82.3,79.0,73.4,72.9,70.2,68.8,30.5,30.3,30.1,29.9,29.7,29.5,29.3,27.8；HRMS(ESI)calcd for C₄₂H₄₃N₅O₁₁K[M+K]⁺832.2591,found 832.2530.

Example 10:

experimental procedure for the synthesis of P12 reference was made to the synthetic procedure for P10 to yield compound P12(32.1mg, 69.1%). [ alpha ] to]_D ²⁵＝-54.8(c 1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ8.74(s,1H),8.71(s,1H),8.14–8.09(m,2H),8.08–8.04(m,2H),7.97–7.92(m,2H),7.69–7.59(m,3H),7.55–7.46(m,4H),7.46–7.40(m,2H),6.79(d,J＝5.0Hz,1H),6.68(dd,J＝6.0,4.9Hz,1H),6.43(t,J＝5.6Hz,1H),5.04–4.99(m,1H),4.95(dd,J＝12.2,3.6Hz,1H),4.81(dd,J＝12.2,4.5Hz,1H),1.41(s,18H)；¹³C NMR(100MHz,Acetone-d₆)δ166.5,165.8,165.7,153.8,152.8,151.4,151.1,146.1,134.6,134.6,134.2,130.8,130.56,130.5,130.5,130.1,130.1,129.8,129.5,129.5,88.3,84.0,81.2,74.5,72.2,64.2,30.5,30.3,30.1,29.9,29.7,29.5,29.3,27.9；HRMS(ESI)calcd for C₄₁H₄₁N₅O₁₁K[M+K]⁺818.2434,found 818.2376.

Example 11:

experimental procedure for the synthesis of P13 reference was made to the synthetic procedure for P10 to yield compound P13(35.0mg, 75.3%). [ alpha ] to]_D ²⁵＝-12.8(c 1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ8.97(s,1H),8.82(s,1H),8.04–7.96(m,2H),7.93–7.86(m,2H),7.75–7.68(m,2H),7.67–7.59(m,1H),7.58–7.46(m,4H),7.39(t,J＝7.8Hz,2H),7.36–7.29(m,2H),6.58(d,J＝9.1Hz,1H),6.45(t,J＝9.3Hz,1H),6.34(t,J＝9.5Hz,1H),5.79(ddd,J＝10.5,9.5,5.6Hz,1H),4.61(dd,J＝11.5,5.6Hz,1H),4.38–4.25(m,1H),1.30(s,18H)；¹³C NMR(100MHz,Acetone-d₆)δ166.2,166.0,165.4,154.1,151.3,150.7,134.6,134.6,134.4,130.4,130.3,130.1,130.0,129.6,129.5,129.4,129.1,129.1,83.8,82.3,74.0,72.3,70.4,66.23,30.5,30.3,30.1,29.9,29.7,29.5,29.3,27.8；HRMS(ESI)calcd for C₄₁H₄₁N₅O₁₁K[M+K]⁺818.2434,found 818.2442.

Example 12:

experimental procedure for the synthesis of P14 reference was made to the synthetic procedure for P10 to yield compound P14(42.2mg, 90.7%). [ alpha ] to]_D ²⁵＝+80.3(c 1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ8.92(s,1H),8.86(s,1H),8.41–8.32(m,2H),7.87–7.80(m,2H),7.78–7.71(m,3H),7.66(ddd,J＝8.3,6.6,1.4Hz,2H),7.56–7.46(m,2H),7.34(q,J＝7.9Hz,4H),6.77(t,J＝9.6Hz,1H),6.53(d,J＝9.2Hz,1H),6.10(dd,J＝10.0,3.4Hz,1H),6.02(dt,J＝3.4,1.4Hz,1H),4.63(dd,J＝13.5,1.2Hz,1H),4.52(dd,J＝13.4,2.1Hz,1H),1.29(s,18H)；¹³C NMR(100MHz,Acetone-d₆)δ166.2,165.8,165.5,154.1,153.0,151.4,150.7,145.2,134.6,134.4,131.0,130.7,130.3,130.3,130.1,129.7,129.5,129.4,129.4,129.3,83.8,73.1,70.1,70.1,68.1,27.8；HRMS(ESI)calcd for C₄₁H₄₁N₅O₁₁K[M+K]⁺818.2434,found 818.2439.

Example 13:

under nitrogen protection, compound S3(20mg,0.054mmol) and I2(52.1mg,0.11mmol) were added to the contents of the activator

Molecular sieves in round bottom flask, dissolved in dry DCM, stirred at room temperature for 0.5h, then NIS (18.2mg,0.08mmol) was added quickly and then placed in a low temperature reactor at-35 deg.C, after reaction for 10min TMSOTf (6.5. mu.L, 0.036mmol) was added slowly, after reaction overnight at this temperature, TCL plate was checked for completion and Et was used₃N quench and spin dry chromatography (PE/EA ═ 3:1) afforded compound P15(32.1mg, 62.6%). [ alpha ] to]_D ²⁵＝+72.6(c 1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ9.22(s,1H),8.09–8.04(m,2H),8.00–7.94(m,2H),7.85–7.79(m,2H),7.77–7.72(m,2H),7.65–7.55(m,2H),7.53–7.41(m,6H),7.35(td,J＝7.8,2.7Hz,4H),6.75(d,J＝9.1Hz,1H),6.55(t,J＝9.3Hz,1H),6.46(t,J＝9.5Hz,1H),6.15(t,J＝9.7Hz,1H),5.04(ddd,J＝10.1,4.2,2.8Hz,1H),4.75–4.63(m,2H),1.37(s,18H)；¹³C NMR(100MHz,Acetone-d₆)δ206.2,166.3,166.0,165.8,165.5,153.9,153.1,151.3,151.1,146.7,134.6,134.5,134.4,134.0,130.6,130.5,130.4,130.3,130.1,129.8,129.7,129.4,129.3,129.3,128.9,83.8,81.4,75.5,74.2,72.3,69.6,63.3,30.4,30.2,30.0,29.8,29.6,29.4,29.2,27.9；HRMS(ESI)calcd for C₄₉H₄₇ClN₅O₁₃[M+H]⁺948.28534,found 948.2882.

Example 14:

experimental procedure for the synthesis of P16 reference was made to the synthetic procedure for P15 to yield compound P16(30.8mg, 68.7%). [ alpha ] to]_D ²⁵＝+18.4(c 1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ9.05(s,1H),7.98–7.90(m,2H),7.76–7.69(m,2H),7.59–7.49(m,2H),7.47–7.40(m,4H),7.38–7.29(m,5H),6.61(d,J＝9.3Hz,1H),6.42(t,J＝9.2Hz,1H),6.24(t,J＝9.2Hz,1H),5.80(s,1H),4.49–4.33(m,3H),3.98(t,J＝9.7Hz,1H),1.39(s,18H)；¹³C NMR(100MHz,Acetone-d₆)δ206.2,166.1,165.7,154.0,153.2,151.4,151.2,146.9,138.4,134.7,134.4,130.8,130.5,130.3,130.3,129.8,129.5,129.5,129.1,128.9,127.2,102.4,84.0,82.3,80.0,73.4,72.7,70.1,68.8,30.5,30.3,30.1,30.0,29.9,29.7,29.5,29.3,28.0；HRMS(ESI)calcd for C₄₂H₄₃ClN₅O₁₁[M+H]⁺828.26421,found 828.2623.

Example 15:

experimental procedure for the synthesis of P17 reference was made to the synthetic procedure for P15 to yield compound P17(36.0mg, 70.2%). [ alpha ] to]_D ²⁵＝+56.1(c 1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ9.04(s,1H),8.31(dt,J＝7.1,1.4Hz,2H),8.02–7.94(m,2H),7.76(ddd,J＝8.4,3.9,1.4Hz,5H),7.69(dd,J＝8.2,6.7Hz,2H),7.66–7.59(m,1H),7.55–7.45(m,4H),7.38–7.27(m,4H),6.72–6.62(m,2H),6.31(dd,J＝3.5,1.2Hz,1H),6.24(dd,J＝9.2,3.5Hz,1H),5.19(td,J＝6.4,1.2Hz,1H),4.68(dd,J＝11.4,6.7Hz,1H),4.58(dd,J＝11.5,6.1Hz,1H),1.37(s,18H)；¹³C NMR(100MHz,Acetone-d₆)δ166.4,166.3,165.7,165.6,154.1,153.2,151.5,151.1,147.2,134.7,134.7,134.5,134.3,131.2,130.9,130.5,130.4,130.4,130.3,130.0,129.9,129.6,129.5,129.4,129.2,83.9,82.9,75.0,72.9,69.9,69.4,63.0,28.0；HRMS(ESI)calcd for C₄₂H₄₂ClN₅O₁₁K[M+K]⁺866.2201,found 866.2170.

Example 16:

experimental procedure for the synthesis of P18 reference was made to the synthetic procedure for P15 to yield compound P18(31.5mg, 71.5%). [ alpha ] to]_D ²⁵＝-37.0(c 1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ8.85(s,1H),8.04(ddt,J＝11.4,6.9,1.4Hz,4H),7.99–7.94(m,2H),7.68–7.60(m,3H),7.51–7.42(m,6H),6.78(d,J＝4.7Hz,1H),6.50(dd,J＝6.0,4.7Hz,1H),6.32(t,J＝5.7Hz,1H),5.03(td,J＝5.5,4.2Hz,1H),4.91(dd,J＝12.0,4.3Hz,1H),4.83(dd,J＝12.0,5.6Hz,1H),1.43(s,18H)；¹³C NMR(100MHz,Acetone-d₆)δ166.5,165.7,165.7,153.5,152.9,151.4,151.3,147.0,134.7,134.6,134.2,131.4,130.7,130.6,130.5,130.4,130.1,129.8,129.5,88.3,84.0,81.5,75.1,72.6,65.0,30.5,30.3,30.2,30.1,29.9,29.7,29.5,29.3,28.0；HRMS(ESI)calcd for C₄₁H₄₀ClN₅O₁₁K[M+K]⁺852.2044,found 852.2041.

Example 17:

experimental procedure for the synthesis of P19 reference was made to the synthetic procedure for P15 to yield compound P19(36.0mg, 81.7%). [ alpha ] to]_D ²⁵＝+32.4(c 1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ9.10(s,1H),8.00–7.94(m,2H),7.91–7.85(m,2H),7.75(dd,J＝8.3,1.4Hz,2H),7.62(t,J＝7.4Hz,1H),7.55–7.46(m,4H),7.37(dt,J＝11.5,7.8Hz,4H),6.54(d,J＝9.2Hz,1H),6.46(t,J＝9.2Hz,1H),6.36(t,J＝9.4Hz,1H),5.81–5.72(m,1H),4.59(dd,J＝11.4,5.6Hz,1H),4.31(t,J＝11.0Hz,1H),1.39(s,18H)；¹³C NMR(100MHz,Acetone-d₆)δ166.2,166.0,165.6,154.0,153.2,151.3,151.3,146.9,134.7,134.6,134.5,130.7,130.5,130.3,130.1,130.0,129.7,129.6,129.6,129.5,129.2,84.0,82.3,73.9,72.0,70.3,66.1,28.0；HRMS(ESI)calcd for C₄₁H₄₀ClN₅O₁₁K[M+K]⁺852.2044,found 852.2029.

Example 18:

experimental procedure for the synthesis of P20 reference was made to the synthetic procedure for P15 to yield compound P20(36.0mg, 81.7%). [ alpha ] to]_D ²⁵＝+152.3(c 1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ9.01(s,1H),8.31–8.25(m,2H),7.82–7.72(m,5H),7.66(dd,J＝8.3,6.7Hz,2H),7.55–7.48(m,2H),7.38–7.30(m,4H),6.61(t,J＝9.6Hz,1H),6.51(d,J＝9.3Hz,1H),6.15(dd,J＝9.8,3.6Hz,1H),6.02(dt,J＝3.5,1.5Hz,1H),4.64(dd,J＝13.6,1.3Hz,1H),4.52(dd,J＝13.5,2.0Hz,1H),1.39(s,18H)；¹³C NMR(100MHz,Acetone-d₆)δ166.3,165.7,165.7,154.1,153.2,151.4,151.2,147.0,134.7,134.5,134.4,131.1,130.8,130.5,130.4,130.3,130.0,129.9,129.5,129.4,129.3,84.0,83.1,72.8,70.1,70.0,68.3,30.5,30.3,30.1,29.9,29.7,29.5,29.3,28.0；HRMS(ESI)calcd for C₄₁H₄₀ClN₅O₁₁K[M+K]⁺852.2044,found 852.2027.

Comparative example: referring to the procedure of example 1, the experimental results for the glycosyl donor and acceptor bases under different reaction conditions are as follows.

(1) Reference to the procedure and parameters in pyrimidine glycosidation example 1: the difference is that the receptor is directly subjected to glycosylation reaction without being activated by BSTFA to obtain a C-O glycosylation product, no C-N product is generated, and the yield is 80-90%. Reference purine glycosidation the procedure and parameters in example 9: except that the molar ratio of RNH to TMSOTf to NIS is 1:1:2, and the yield is 20-30%.

Comparative example 1:

under nitrogen protection, compound S1(20mg,0.064mmol) and I1(53.2mg,0.096mmol) were added to the contents of the flask

Dried DCM (1.0mL) was added to a round bottom flask of molecular sieves and stirred at room temperature for 0.5h, then NIS (29mg,0.13mmol) was added quickly and then placed in a low temperature reactor at-35 deg.C, after reaction for 10min TMSOTf (7.0uL,0.038mmol) was added slowly and reacted at this temperature overnight to give Compound A (40.0mg, 82.5%). [ alpha ] to]_D ²⁵＝-5.5(c 1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ8.56(d,J＝5.6Hz,1H),7.49(d,J＝5.6Hz,1H),6.24(d,J＝8.2Hz,1H),5.41(t,J＝9.5Hz,1H),5.26–5.11(m,2H),4.32–4.23(m,1H),4.14–4.06(m,2H),2.02(s,3H),1.98(s,3H),1.97(s,3H),1.96(s,3H),1.56(s,18H)；¹³C NMR(100MHz,Acetone-d₆)δ170.7,170.3,170.0,169.6,163.7,161.2,150.7,108.4,95.4,85.2,73.5,73.2,71.6,69.1,62.5,28.0,27.9,20.7,20.6,20.6；HRMS(ESI)calcd for C₂₈H₃₉N₃O₁₄Na[M+Na]⁺664.2432,found 664.2443.

Comparative example 2:

experimental procedure for synthesis B compound B (50.4mg, 88.2%) was obtained by reference to the synthesis procedure for a. [ alpha ] to]_D ²⁵＝+34.3(c 1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ8.51(d,J＝5.6Hz,1H),8.06–8.00(m,2H),7.95(ddd,J＝10.0,8.4,1.4Hz,4H),7.89–7.83(m,2H),7.66–7.52(m,4H),7.51–7.36(m,9H),6.68(d,J＝8.1Hz,1H),6.20(t,J＝9.4Hz,1H),5.93–5.83(m,2H),4.72–4.56(m,3H),1.50(s,18H)；¹³C NMR(100MHz,Acetone-d₆)δ166.4,166.2,165.9,165.5,163.7,161.2,161.1,150.5,134.5,134.5,134.4,134.1,130.8,130.5,130.5,130.5,130.4,130.1,130.0,129.5,129.4,129.4,108.3,95.8,85.1,74.2,73.5,72.3,70.4,63.6,27.9；HRMS(ESI)calcd for C₄₈H₄₇N₃O₁₄K[M+K]⁺928.2690,found 928.2695.

Comparative example 3:

experimental procedure for Synthesis C reference was made to the synthetic procedure for A to give Compound C (41.8mg, 84.5%). [ alpha ] to]_D ²⁵＝+44.3(c 1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ8.51(d,J＝5.6Hz,1H),8.00–7.93(m,2H),7.93–7.86(m,2H),7.58–7.48(m,2H),7.47–7.34(m,7H),7.33–7.26(m,3H),6.59(d,J＝8.0Hz,1H),5.96(t,J＝9.4Hz,1H),5.81–5.68(m,2H),4.39(dd,J＝9.7,4.3Hz,1H),4.27(t,J＝9.3Hz,1H),4.08–3.90(m,2H),1.52(s,16H)；¹³C NMR(100MHz,Acetone-d₆)δ166.0,165.5,163.6,161.2,161.1,150.5,138.4,134.4,134.3,130.4,130.0,129.7,129.4,128.9,127.2,108.2,102.2,96.1,85.1,79.1,73.2,73.0,68.9,67.9,27.9；HRMS(ESI)calcd for C₄₈H₄₇N₃O₁₄Na[M+Na]⁺808.2478,found 808.2477.

Comparative example 4:

experimental procedure for Synthesis D reference was made to the synthetic procedure for A to give Compound D (53.7mg, 93.9%). [ alpha ] to]_D ²⁵＝+85.9(c 1.0,CHCl₃)；¹H NMR(400MHz,Acetone-d₆)δ8.53(d,J＝5.6Hz,1H),8.17–8.10(m,2H),8.03–7.97(m,2H),7.96–7.91(m,2H),7.83–7.78(m,2H),7.75–7.70(m,1H),7.64–7.57(m,3H),7.56–7.46(m,5H),7.40(t,J＝7.8Hz,2H),7.36–7.30(m,2H),6.68(d,J＝8.2Hz,1H),6.17(dd,J＝3.5,1.2Hz,1H),6.12(dd,J＝10.3,8.1Hz,1H),6.02(dd,J＝10.3,3.4Hz,1H),4.96–4.88(m,1H),4.66(dd,J＝11.3,6.9Hz,1H),4.55(dd,J＝11.3,6.1Hz,1H),1.50(s,18H)；¹³C NMR(100MHz,Acetone-d₆)δ166.3,166.3,165.7,165.7,163.8,161.2,161.2,150.6,134.7,134.4,134.41,134.7,130.7,130.6,130.4,130.4,130.3,130.2,130.0,129.8,129.8,129.5,129.4,129.4,108.3,96.0,85.1,72.8,72.8,70.3,69.5,62.8,27.9；HRMS(ESI)calcd for C₄₈H₄₇N₃O₁₄Na[M+Na]⁺902.3058,found 902.3065.

Comparative example 5:

under nitrogen, compounds S2(20mg,0.060mmol) and I3(48.8mg,0.072mmol) were dissolved in dry DCM and stirred at room temperature for 0.5h, NIS (26.8mg,0.12mmol) was added rapidly, placed in a low temperature reactor at-35 deg.C, TMSOTf (10.8. mu.L, 0.060mmol) was added slowly after reaction for 10min, and after completion of the reaction was checked by overnight reaction on TCL plates at this temperature, Et was used₃N quench and spin dry chromatography (PE/EA ═ 4:1) afforded compound P10(9.8mg, 21.0%).

Comparative example 6:

experimental procedure for the synthesis of P12 reference was made to the synthetic procedure for P10 in comparative example 5 to yield compound P12(11.9mg, 25.6%).

Comparative example 7:

experimental procedure for the synthesis of P15 reference was made to the synthetic procedure for P10 in comparative example 5 to give compound P15(14.4mg, 28.1%).

Comparative example 7:

experimental procedure for the synthesis of P18 reference was made to the synthetic procedure for P10 in comparative example 5 to give compound P18(9.5mg, 21.5%).

Claims (7)

1. A method for glycosylation of a base by taking an o-alkynyl phenol ether as a leaving group is characterized in that a glycosylation donor shown in a formula I and a glycosylation receptor shown in a formula S are subjected to glycosylation reaction under the action of an accelerant in the presence of a drying agent in an inert gas environment to obtain a glycosylation product shown in a formula P,

wherein RNH is a basic group compound, Gly is glycosyl with one or more hydroxyl groups on a glycosyl ring protected by a protecting group, and the temperature of glycosylation reaction is-50 to-20 ℃;

the base is a pyrimidine or a purine,

when the base is a pyrimidine, the promoters are BSTFA, NIS and TMSOTf;

when the base is a purine, the enhancers are NIS and TMSOTf.

2. The method of claim 1, wherein the base-like compound RNH is selected from any one of compounds represented by the following formulas S1 to S3,

wherein R' is an acyl protecting group.

3. The method of claim 2, wherein the acyl-based protecting group is selected from the group consisting of Bz, Cbz and Boc.

4. The method of base glycosidation according to claim 2, wherein said glycosidation donor is selected from any of the compounds shown below,

。

5. the method of claim 4, wherein the base is a purine, the molar ratio of RNH to TMSOTf and NIS is 1:0.1:1 to 1:1:2, the molar ratio of the glycosylation donor to RNH is 1:1 to 5:1, and the glycosylation reaction is carried out for 3 to 15 hours.

6. The method of claim 4, wherein the base is a pyrimidine, the molar ratio of RNH to TMSOTf, NIS to BSTFA is 1:0.1:1:1 to 1:1:2:4, the molar ratio of the glycosylation donor to RNH is 1:1 to 5:1, and the time of the glycosylation reaction is 3 to 15 hours.

7. The method of base glycosidation according to any of claims 1 to 6, wherein the inert gas is selected from the group consisting of argon or nitrogen; the drying agent is selected from acid-washed molecular sieves.