Method for preparing On-DNA sulfinic acid structure compound and application of On-DNA sulfinic acid structure compound

Abstract

The invention belongs to the field of preparation of medical intermediates, and particularly relates to a preparation method of an On-DNA sulfinic acid structure compound. The method for preparing the On-DNA sulfinic acid structure compound comprises a step of oxidizing On-DNA thiol (phenol) into the On-DNA sulfinic acid structure compound under alkaline and heating or illumination conditions, a solvent used in the reaction step is a mixed solution of water, dimethyl sulfoxide, N, N-dimethylformamide, acetonitrile, ethanol or tetrahydrofuran, the reaction temperature is not more than 80 DEG C, and the reaction time is not more than 2 hours. And the illumination wavelength ranges from 365 nanometers to 535 nanometers. The On-DNA sulfinic acid structure compound obtained in the invention can be directly used for constructing a structure compound with a sulfonyl group, the reaction condition is mild and convenient, a new method and a new idea are provided for construction of a DNA coding compound library, and the On-DNA sulfinic acid structure compound has a very good application prospect in lead drug development.

Description

Technical Field

[0001] The present application belongs to the field of preparation of pharmaceutical intermediates, and specifically relates to an On-DNA sulfinic acid structure compound or a method for preparing a sulfinic acid structure compound. Background Art

[0002] Many pharmacophores contain sulfonyl structures, such as 11β-HSD1 inhibitors (11β-HSD1-IN-11) and oxazolidinone fungicides (Oxycarboxin) with β-ketosulfone structures; neuroprotectants ((E)-1-methoxy-2-(styrylsulfonyl)benzene), Nrf2 agonists ((E)-4-(3-(4-((2-(3′-fluoropyridin-2′-yl)vinyl)sulfonyl)phenoxy)propyl)morpholine), and microtubule inhibitors ((E)-1,2,3-trimethoxy-5-(styrylsulfonyl)benzene) with alkenylsulfone structures; bicalutamide (Bicalutamide) with β-hydroxysulfone structures, and antifungal drugs SCH42427 and SSY726.

[0003] In the prior art, a key intermediate used in the preparation of the above-mentioned compounds containing sulfonyl structures is sulfinic acid. For example, Long-Yong Xie (Photosensitizer-free synthesis of β-keto sulfones via visible-light-induced oxysulfonylation of alkenes with sulfonic acids, Organic & Biomolecular Chemistry, DOI: 10.1039 / d1ob00552a) et al. synthesized a compound having the following β-keto sulfone structure using sulfinic acid and olefins under blue light irradiation; Yangfeng Li and Yizhou Li et al. synthesized vinyl sulfone using sodium sulfinate and olefins under iodine catalysis (Development of on-DNA vinyl sulfone synthesis for DNA-encoded chemical library, DOI: 10.1039 / D2QO00881E); Patent document CN118257005A discloses a method for generating β-hydroxy sulfone compounds by reacting olefin compounds with sodium sulfinate. This shows that sulfinic acid is an important intermediate for the synthesis of many pharmaceutically effective structures.

[0004] Traditional pharmacodynamic compound screening is to synthesize a single small molecule compound with a pharmacodynamic group using organic synthesis methods. DNA-Encoded Chemical Library (DEL) technology is an emerging small molecule screening technology that can quickly construct a large-scale compound library based on combinatorial chemistry principles. With the deepening application of technologies such as artificial intelligence and deep learning in various industries, the use of AI technology to accelerate new drug discovery has become more and more popular. Since AI technology is often data-driven, that is, to capture the potential laws in the data through mathematical models, in order to realize the implementation of AI technology in the biomedical field, it is necessary to combine scenarios that can quickly generate and accumulate high-quality biological big data. The massive normalized and standardized experimental data generated by DEL technology with tens of billions or even hundreds of billions of chemical spaces is an important data source for training reliable AI models. Using AI algorithms to deeply mine DEL data, especially systematically analyzing the affinity performance of molecules with similar structures on interrelated targets, it is more likely to discover small molecules with unique structural properties, so as to accelerate the discovery of lead compounds and even the entire drug discovery process. The new model of DEL+AI provides unlimited opportunities and possible space for the discovery of small molecule drugs in the future.

[0005] However, after extensive literature searches, the inventors of the present application have not found a method for quickly constructing a large number of sulfinic acid structure compounds at one time, which cannot meet the screening needs of the DEL+AI model for effective compounds with sulfonyl groups. Summary of the invention

[0006] In order to solve the need of DEL+AI mode screening of pharmacological compounds, the present application first provides a method for constructing On-DNA sulfinic acid structure compounds based on DEL technology. The specific method includes the following reaction wherein R1 is selected from substituted and unsubstituted alkyl, benzyl, aryl or heteroaryl, and the substituent of the substituted R1 is halogen, alkyl or alkoxy; Preferably, the substituted or unsubstituted alkyl of R1 is a C2-C3 alkyl; the substituted or unsubstituted aryl of R1 is a hexacyclic aryl; the heteroatom of the substituted or unsubstituted heterocyclic aryl of R1 is N, and the number of the heteroatoms is 1; the substituent of R1 is fluorine, C1 alkyl or alkoxy.

[0007] The base is sodium hydroxide, potassium hydroxide, sodium carbonate or sodium bicarbonate, potassium carbonate or potassium bicarbonate, N,N-diisopropylethylamine, bicyclic amidine, triethylamine, boric acid buffer with a pH of 9.5 or triethylenediamine, and the concentration of the base in the reaction system is 0.1mM to 2mM; X is a proton corresponding to the base and bound to the sulfinic acid group, for example, if the base is sodium hydroxide, then X is sodium; if the base is potassium hydroxide, then X is potassium; and so on. During the experiment, the inventors of the present application found that weak bases make the reaction conditions milder, and strong base carbonate salts are more conducive to the purification of subsequent products because carbonate groups will be converted into carbon dioxide and water during the reaction process, and are more conducive to the requirements of being a medicinal compound.

[0008] The reaction solvent includes water and an organic solvent, wherein the organic solvent is dimethyl sulfoxide, N,N-dimethylformamide, acetonitrile, ethanol or tetrahydrofuran; the ratio of water to organic solvent is 3:97-3:7; the applicant found in the test process that the effect of the mixed solvent of DMSO and DMF with water is significantly better than that of the mixed solvent of MeCN, EtOH, THF and 1,4-dioxane with water. When the ratio of water to DMSO is in the range of 3:97 to 3:7 and other reaction conditions are the same, the conversion rate of the sulfinic acid structure compound gradually decreases.

[0009] The temperature is not higher than 80°C. During the experiment, the inventors of the present application, under the same other conditions, simply raised the reaction temperature to 80°C for one hour, the integrity of the DNA was good, and the conversion rate of the On-DNA sulfinic acid structure compound was nearly 70%. Such a conversion rate was significantly higher than the conversion rate of the reaction at 60°C for one hour. Moreover, within the reaction range of room temperature to 80°C, the conversion rate of the On-DNA sulfinic acid structure compound still showed a gradually increasing trend.

[0010] The light source is 365 nanometers to 535 nanometers; the reaction can be carried out without the light source during the test, and the yield can reach 35%; the conversion of the On-DNA sulfinic acid structure compound first increases and then decreases when the wavelength of the light source is within the range of 365 nanometers to 535 nanometers, so the preferred wavelength for the reaction is 365 nanometers to 405 nanometers. The light source irradiation can effectively reduce the reaction temperature and the reaction conditions are milder. For example, in the absence of light source irradiation, the reaction temperature is 80°C and the reaction time is 1 hour, and the conversion rate of the On-DNA sulfinic acid structure compound is 69%. If the light source irradiation is used under the same conditions as the other conditions of the above reaction, the reaction at room temperature for 15 minutes can reach a conversion rate of 77%.

[0011] Preferably, the reaction further requires the addition of dithiothreitol, preferably at a final concentration of 0.1-1.0 mmol / L, the final concentration being the concentration in the reaction system. Under the same other conditions, the addition of DTT to the reaction system effectively increases the conversion rate of the On-DNA sulfinic acid structure compound by 20%.

[0012] It is well known in the art that the reaction for constructing a DNA-encoded compound library needs to have a high yield, and can quickly synthesize compounds with the same pharmacophore at a hundred or even a thousand times the order of magnitude of small molecule organic chemistry at one time. After obtaining the On-DNA sulfinic acid structure compound, the next step can be directly carried out to construct a structural compound with a sulfonyl group, such as the β-ketosulfone, alkenylsulfone and other structural compounds mentioned in the prior art. Using the reaction conditions of the present invention, the obtained oligonucleic acid-sulfinic acid structure compound not only has a high yield, but also has good DNA integrity in the oligonucleic acid-sulfinic acid structure compound. The integrity of the oligonucleic acid-sulfinic acid structure compound product obtained in the present invention can be confirmed from liquid chromatography-mass spectrometry. The present invention enriches the chemical reaction types for synthesizing encoded compound libraries on DNA, provides a new method for the construction of synthetic DNA encoded compound libraries, and has very good application prospects in lead drug development. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1-19 : Liquid chromatography mass spectra of oligonucleic acid-sulfinic acid structure compounds 2a-2s; Fig. 20 : Liquid chromatography mass spectrum of oligonucleic acid-vinyl sulfone structure compound 3m obtained in application example; DETAILED DESCRIPTION

[0014] DIEA: N,N-diisopropylethylamine; DBU: abbreviation of bicyclic amidine, also known as bicyclic amidine, chemical name is 1,8-diazabicyclo (5,4,0)-7-undecene; TEA: triethylamine pH = 9.5 borate buffer; DABCO: triethylenediamine; DTT: dithiothreitol; DMSO: dimethyl sulfoxide; MeCN: acetonitrile; DMF: N,N-dimethylformamide; EtOH: ethanol; 1,4-dioxane: 1,4-dioxane; THF: tetrahydrofuran; Oligonucleic acid-thiol (phenol) compounds are direct raw materials for preparing On-DNA sulfinic acid structure compounds. The preparation method of oligonucleic acid-thiol (phenol) compounds is as follows: (1) HP-Linker-NHFmoc is prepared from HP-NH2 (commercially available reagent), (2) HP-Linker-NH2 is then prepared from HP-Linker-NHFmoc, and (3) in the third step, HP-Linker-NH2 is converted into oligonucleic acid-thiol (phenol) compounds. This application uses exactly the same method as application number 2024119940775 to prepare HP-Linker-NH2 and the structure of oligonucleic acid-thiol (phenol) compounds, so the structure confirmation diagrams of HP-NH2, HP-Linker-NHFmoc, HP-Linker-NH2, and oligonucleic acid-thiol (phenol) compounds 1a-1s are omitted.

[0015] 1. Synthesis of oligonucleotide-Linker-NH2 raw material (HP-Linker-NH2) 1.1 Preparation of HP-Linker-NHFmoc from HP-NH2 (commercially available reagent) has the following reaction equation: Specifically, 100.0 nanomoles of HP-NH2 (commercially available product) was dissolved in deionized water to prepare a 1.0 mmol / L solution (100.0 microliters, 100.0 nanomoles, 1.0 equivalent). 40.0 equivalents of S1 (commercially available product) in DMSO solution (concentration: 200.0 mmol / L), 250.0 equivalents of sodium tetraborate (Na2B4O7) buffer solution with pH = 9.5 (concentration: 250.0 mmol / L), and 40.0 equivalents of 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (DMT-MM) aqueous solution (concentration: 200.0 mmol / L) were mixed, and the mixture was fully mixed with a vortex oscillator. The above mixture was then added to the HP-NH2 solution, mixed evenly, and reacted at 4°C for 1 hour. After the reaction was completed, 10% of the total volume of 5.0 mol / L sodium chloride solution was added to the reaction solution. Then, continue to add anhydrous ethanol 3 times the total volume, shake evenly, and place the reaction solution in a -80°C refrigerator for 2 hours. After that, centrifuge at 4000.0rpm for half an hour and pour off the supernatant. The remaining precipitate is dissolved in deionized water to obtain a solution of HP-Linker-NHFmoc. HP-Linker-NHFmoc is detected by liquid chromatography-mass spectrometry, and its molecular weight is 5406.

[0016] 1.2 Preparation of HP-Linker-NH2 from HP-Linker-NHFmoc: 100.0 nanomoles of HP-Linker-NHFmoc were dissolved in deionized water to prepare 1.0 mmol / L (100.0 microliters, 100.0 nanomoles, 1.0 equivalents), and 36.0 microliters of 10% piperidine aqueous solution were added thereto. The two were mixed evenly and reacted at room temperature for 1 hour. After the reaction was completed, a 5.0 mol / L sodium chloride solution with a total volume of 10% was added to the reaction solution. Then, 3 times the total volume of anhydrous ethanol was added, and after oscillation, the reaction was placed in a refrigerator at -80°C and frozen for 2 hours. After that, centrifuged at a speed of 4000.0rpm for half an hour, and the supernatant was poured out. The remaining precipitate was dissolved in deionized water to obtain a solution of oligonucleotide-Linker-NH2 (HP-Linker-NH2). HP-Linker-NH2 was detected by liquid chromatography-mass spectrometry, and its molecular weight was 5184.

[0017] 2. Sources or synthesis methods of raw materials used to prepare oligonucleic acid-thiol (phenol) compounds: The present application prepares oligonucleic acid-thiol (phenol) compounds by reacting the following molecules with HP-Linker-NH2: Except for S2j, S2l, and S2p, which are homemade, the rest are commercially available. The reaction equations for preparing S2j, S2l, and S2p are: Where a is HCl, NaNO2, Na2S.9H2O, S8, NaOH, 0℃ to rt; where b is DMSO, 80℃.

[0018] A more specific method for preparing S2j, S2l, and S2p is as follows: 0.3 g (1.3 eq.) of sodium nitrite (NaNO2) is dissolved in 1.0 ml of ice-cooled aqueous solution, and then dropwise added to a mixed solution of 1.0 ml of aqueous solution of 3.3 mmol (1.0 eq.) of aminocarboxylic acid and 0.6 ml of concentrated hydrochloric acid (HCl) at 0°C, and the mixture is stirred at 5°C for 30 minutes. Then, 1.04 g (1.3 eq.) of sodium sulfide nine hydrate (Na2S.9H2O) in 1.0 ml of cooled aqueous solution, 138.0 mg (0.16 eq.) of sulfur (S8), and 0.3 ml of 10.0 mol / L sodium hydroxide (NaOH) aqueous solution are added in sequence. The mixture is stirred at room temperature for 2 hours, and then acidified to pH 2 with hydrochloric acid. The resulting precipitate is collected by filtration, washed with water, dried under high vacuum, and can be entered into the next step without further purification.

[0019] The solid obtained in the previous step was dissolved in 5.0 ml of DMSO and stirred at 80°C overnight. Purification by reverse phase column chromatography gave the corresponding yellow solid disulfide.

[0020] S2j Preparation of: Disulfide S2j (131.0 mg, 23%) was obtained from 3-amino-5-methylbenzoic acid (500.0 mg, 3.31 mmol) as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ13.11(s,2H),7.90–7.84(m,2H),7.69–7.66(m,2H),7.61–7.56(m,2H),2.35(s,6H).MS(ESI-MS)m / z:333[MH] + . S2 Preparation of: Disulfide (S21, 197.0 mg, 35%) was obtained from 3-amino-5-fluorobenzoic acid (500.0 mg, 3.22 mmol) as a yellow solid. 1 H NMR(400MHz, DMSO-d6)δ13.53(s,2H),7.91(t,J=1.6Hz,2H),7.72(dt,J=8.8,2.1Hz,2H),7.62–7.54(m,2H).MS(ESI-MS)m / z:341[MH] + . S2p Preparation of: Disulfide (S2p, 134.0 mg, 24%) was obtained from 4-amino-3-fluorobenzoic acid (500.0 mg, 3.22 mmol) as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ13.40(s,2H),7.91–7.64(m,6H).MS(ESI-MS)m / z:341[MH] + . 3. Preparation of oligonucleic acid-thiol (phenol) compounds 3.1 Preparation of oligonucleic acid-thiol (phenol) compounds from HP-Linker-NH2 3.1.1 Synthesis of oligonucleic acid-thiol (phenol) compound raw material (1a) prepared from HP-Linker-NH2 50.0 nanomoles of HP-Linker-NH2 were dissolved in deionized water to prepare 1.0 mmol / L (50.0 μL, 50.0 nanomoles, final concentration 0.2 mmol / L). Mix the DMSO solution of disulfide S2a (commercial product) (200.0 mmol / L, 50.0 μL, final concentration of 40.0 mmol / L), EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 1,200.0 mmol / L, 50.0 μL, final concentration of 240.0 mmol / L), HOAt (1-hydroxy-7-azobenzotriazole, 240.0 mmol / L, 50.0 μL, final concentration of 48.0 mmol / L) and DIEA (N,N-diisopropylethylamine, 1,200.0 mmol / L, 50.0 μL, final concentration of 240.0 mmol / L), and mix the mixture thoroughly with a vortex oscillator. Then add the above mixture to the solution of HP-Linker-NH2, mix well and react at 25°C for 1 hour. After the reaction is completed, add 10% of the total volume of 5.0 mol / L sodium chloride solution to the reaction solution. Then, continue to add 3 times the total volume of anhydrous ethanol, shake evenly, and place the reaction solution in a -80°C refrigerator for 2 hours. After that, centrifuge at 4000.0 rpm for half an hour, and pour out the supernatant. The remaining precipitate is treated with DTT to cleave the disulfide bond (patent: CN 114853822B), and then precipitated with sodium chloride / ethanol and dissolved in deionized water to obtain a solution of 1a, whose molecular weight is 5272.6 and the yield is 76%.

[0021] The inventive team of this application adopted the method for preparing raw material 1a to synthesize other oligonucleic acid-thiol (phenol) compounds (1b-g, 1j-1q).

[0022] 3.1.2 Synthesis of oligonucleic acid-thiol (phenol) compound raw materials (1h) prepared from HP-Linker-NH2 Dissolve 50.0 nanomoles of HP-Linker-NH2 in deionized water to make 1.0 mmol / L (50.0 microliters, 50.0 nanomoles, final concentration of 0.2 mmol / L). Mix sulfide S2h (commercial product) (200.0 mmol / L, 50.0 microliters, final concentration of 40.0 mmol / L), EDCI (1,200.0 mmol / L, 50.0 microliters, final concentration of 240.0 mmol / L), HOAt (240.0 mmol / L, 50.0 microliters, final concentration of 48.0 mmol / L) and DIEA (1,200.0 mmol / L, 50.0 microliters, final concentration of 240.0 mmol / L) in DMSO, and mix the mixture thoroughly with a vortex oscillator. Then add the above mixture to the HP-Linker-NH2 solution, mix well and react at 25°C for 1 hour. After the reaction is completed, add 10% of the total volume of 5.0 mol / L sodium chloride solution to the reaction solution. Then, continue to add 3 times the total volume of anhydrous ethanol, shake evenly, and place the reaction solution in a -80°C refrigerator for 2 hours. After that, centrifuge at 4000.0rpm for half an hour and pour off the supernatant. The remaining precipitate is dissolved in deionized water to obtain a 1h solution, the molecular weight of which is 5338.9 and the yield is >99%.

[0023] The inventive team of this application adopted the method for preparing raw material 1h to synthesize other oligonucleic acid-thiol (phenol) compounds (1i, 1r-1s).

[0024] The oligonucleic acid-thiol (phenol) compounds 1a-1s prepared through the above preparation process have the following structures: The structure confirmation map has been disclosed in application number 2024119940775 and is omitted here.

[0025] Example 1-31 Taking the following reaction as an example, the reaction conditions were optimized Wherein X is the proton bound to the sulfinate group corresponding to the base. Specific reaction conditions and reaction results are shown in Table 1.

[0026] Examples 1-3 were conducted under fluorescent lamps in the laboratory without any additional light source. Comparing with Examples 1-4, we can see that although heating can achieve the conversion of raw materials into target products, light sources can effectively improve the conversion rate.

[0027] By comparing Examples 5-8, we can see that when other reaction conditions remain unchanged, the conversion of On-DNA sulfinic acid structure compounds first increases and then decreases when the wavelength of the light source is in the range of 365 nm to 535 nm, so the preferred reaction wavelength is 365 nm to 405 nm. Compared with the condition without light source heating, the light source can significantly shorten the reaction time, increase the conversion rate, and make the reaction conditions milder and more convenient.

[0028] By comparing Examples 6 and 9, we can see that when other reaction conditions remain unchanged, without adding DTT, the conversion rate of On-DNA sulfinic acid structure compound 2m will decrease and the impurity content of the raw material dimerization will increase. Therefore, it is preferred to add DTT at a final concentration of 0.1-1.0 mmol / L.

[0029] By comparing Examples 6 and 10-17, we can see that when other reaction conditions remain unchanged, different bases have little effect on the conversion rate of 2m. Considering that carbonates or bicarbonates are weak bases, the reaction conditions are milder and they decompose into CO2 and H2O when exposed to acid, which is more environmentally friendly. Therefore, we choose carbonates or bicarbonates as the base additives for the reaction.

[0030] Comparing Examples 12, 18-21, we can see that when the reagent amount of NaHCO3 is reduced from 2 mmol / L to 0.1 mmol / L under the condition that other reaction conditions remain unchanged, there is no difference in the 2m conversion rate, but when NaHCO3 is removed, the 2m conversion rate drops sharply, so it is preferred to add NaHCO3 to a final concentration of 0.1-2.0 mmol / L.

[0031] By comparing Examples 20, 22-26, we can see that the effect of solvent on the conversion rate of On-DNA sulfinic acid structure compound 2m is that the effect of the mixed solvent of water with DMSO and DMF is significantly better than the mixed solvent of water with MeCN, EtOH, THF and 1,4-dioxane.

[0032] Comparing Examples 20, 27-28, we can see that the effect of the volume ratio of water to organic solvent on the conversion rate of On-DNA sulfinic acid structure compound 2m is 3:97>3:17>3:7. Based on this experimental result, the applicant believes that the ratio of water to organic solvent used in the reaction is 3:97-3:7.

[0033] By comparing Examples 28-31, we can see that when other reaction conditions remain unchanged, when the final concentration of 1m is 0.05 mmol / L, the conversion rate of 2m is better than others. The inventors of the present application found in the experimental process that even in the range of 0.005 mmol / L to 0.2 mmol / L, in order to keep the conversion rate unchanged, for example, to achieve a conversion rate of 91%, it is sufficient to increase the amount of other materials, but for cost-effectiveness, 0.01 to 0.2 mmol / L is preferred, and 0.02 to 0.2 mmol / L is more preferred.

[0034] Example 32 Synthesis of oligonucleic acid-sulfinic acid structure compound (2a) from oligonucleic acid-thiol (phenol) compound raw material (1a) Oligonucleic acid-thiol (phenol) compounds (1a, 5.0 nanomoles, 1.0 equivalents, 2.0 mmol / L aqueous solution, 2.5 μL, 0.05 mmol / L final concentration), NaHCO3 (20.0 mmol / L aqueous solution, 0.5 μL, 0.1 mmol / L final concentration), DTT (dithiothreitol, 5.0 mmol / L DMSO solution, 10.0 μL, 0.5 mmol / L final concentration) and 87.0 μL DMSO were added to a 0.6 ml centrifuge tube. The mixture was thoroughly mixed by vortexing, and then shaken and irradiated (405 nm, 7 W UV lamp) at 25° C. for 15 minutes. After the reaction, 10% of the total volume of 5.0 mol / L sodium chloride solution and 3 times of anhydrous ethanol were added to the reaction solution. After being shaken evenly, it was placed in a refrigerator at -80°C for 2 hours, and then high-speed refrigerated centrifugation (4°C, 12000 rpm, 15 minutes) was performed. The supernatant was poured out and the remaining precipitate was the product oligonucleic acid-sulfinic acid structure compound (2a). The molecular weight was 5326.6 and the conversion rate was >99% when detected by liquid chromatography-mass spectrometry.

[0035] Embodiment 33-50 Different from Example 32, oligonucleic acid-thiol (phenol) compounds (1b-1s) were used as raw materials to synthesize other representative oligonucleic acid-sulfinic acid structure compounds (2b-2s), and their specific structures are as follows. Figure 1-19 .

[0036] The applicant would like to explain that the above list is only an example, because we can understand that even if each oligonucleic acid-thiol (phenol) compound raw material is listed in the table, it can form On-DNA sulfinic acid structure compounds with different structures through this reaction, and therefore it cannot be exhausted.

[0037] Application Examples In a 0.6 ml centrifuge tube, oligonucleic acid-sulfinic acid compound (2m, 5.0 nanomoles, 1.0 equivalents, 1.0 mmol / L aqueous solution, 5.0 μl, 0.33 mmol / L final concentration), S3 (100.0 mmol / L ethanol solution, 5.0 μl, 33.3 mmol / L final concentration) and sodium dihydrogen phosphate buffer solution (5.0 μl) at pH = 3.6 were added. The mixture was thoroughly mixed by vortex oscillation and then oscillated at 25°C for 1 hour. After the reaction was completed, 10% of the total volume of 5.0 mol / L sodium chloride solution and 3 times of anhydrous ethanol were added to the reaction solution. After oscillation, it was placed in a -80°C refrigerator for 2 hours, and then high-speed refrigerated centrifugation (4°C, 12,000 rpm, 15 minutes) was performed, and the supernatant was discarded. The remaining precipitate was the product oligonucleic acid-vinyl sulfone structure compound (3m, Fig. 20 ).

[0038] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make modifications to the present embodiment without any creative contribution as needed, but such modifications are protected by the patent law as long as they are within the scope of the claims of the present application.

Claims

1. A method for preparing an On-DNA sulfinic acid structure compound, comprising the reaction steps of preparing an On-DNA sulfinic acid structure compound from an On-DNA thiol or an On-DNA thiophenol, the reaction equation is as follows The X is a proton bound to the sulfinate group corresponding to the base, R 1 is selected from substituted or unsubstituted alkyl, benzyl, aryl or heteroaryl, wherein the substituted R 1 The substituent is halogen, alkyl or alkoxy; The base is sodium hydroxide, potassium hydroxide, sodium carbonate or sodium bicarbonate, potassium carbonate or potassium bicarbonate, N,N-diisopropylethylamine, bicyclic amidine, triethylamine, boric acid buffer with a pH of 9.5 or triethylenediamine; the temperature is not higher than 80°C.

2. The method for preparing an On-DNA sulfinic acid structure compound according to claim 1, characterized in that: The R 1 is a substituted or unsubstituted C2-C3 alkyl group; said R 1 is a substituted or unsubstituted aryl group, a six-membered aryl group; said R 1 The heteroatom in the substituted or unsubstituted heterocyclic aromatic group is N, and the number of the heteroatom is 1; the R 1 The substituent is fluorine or C1 alkyl or alkoxy.

3. The method for preparing an On-DNA sulfinic acid structure compound according to claim 1, characterized in that: The concentration of the base in the reaction system is 0.1-2.0 mmol / L final concentration.

4. The method for preparing an On-DNA sulfinic acid structure compound according to claim 1, characterized in that: The solvent includes water and an organic solvent, and the organic solvent is dimethyl sulfoxide, N,N-dimethylformamide, acetonitrile, ethanol or tetrahydrofuran.

5. The method for preparing an On-DNA sulfinic acid structure compound according to claim 4, characterized in that: The ratio of water to organic solvent is 3:97-3:

7.

6. The method for preparing an On-DNA sulfinic acid structure compound according to claim 1, characterized in that: The reaction step of preparing the On-DNA sulfinic acid structure compound from On-DNA thiol or On-DNA thiophenol can also be irradiated by a light source with a wavelength of 365 nm to 535 nm.

7. The method for preparing an On-DNA sulfinic acid structure compound according to claim 6, characterized in that: The light source is 365 to 405 nanometers.

8. The method for preparing an On-DNA sulfinic acid structure compound according to claim 1, characterized in that: The reaction also requires the addition of dithiothreitol.

9. The method for preparing an On-DNA sulfinic acid structure compound according to claim 8, characterized in that: The concentration of the dithiothreitol is 0.1-1.0 mmol / L final concentration.

10. Use the method for preparing an On-DNA sulfinic acid structure compound according to any one of claims 1 to 9 for preparing a sulfinic acid structure compound.

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