A method for preparing On-DNA sulfinic acid structure compound and its application

Abstract

The present application belongs to the field of preparation of pharmaceutical intermediates, and specifically relates to a method for preparing an On-DNA sulfinic acid structure compound. The method for preparing an On-DNA sulfinic acid structure compound includes the steps of oxidizing an On-DNA thiol (phenol) to an On-DNA sulfinic acid structure compound under alkaline and heating or light conditions, and the solvent used in this reaction step is a mixed solution of water and dimethyl sulfoxide, N,N-dimethylformamide, acetonitrile, ethanol or tetrahydrofuran. The reaction temperature does not exceed 80°C, and the illumination wavelength is 365 nanometers to 535 nanometers. The On-DNA sulfinic acid structure compound obtained in this application can be directly used to construct a structural compound having a sulfonyl group. The reaction conditions are mild and convenient, which provides a new method and new idea for the construction of a DNA-encoded compound library and has very good application prospects in lead drug development.

Description

Technical Field

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

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

[0003] In the prior art, a key intermediate used in the preparation of the above-mentioned sulfonyl-containing compounds is sulfinic acid. For example, Long-Yong Xie et al. (Photosensitizer-free synthesis of β-keto sulfones via visible-light-induced oxysulfonylation of alkenes with sulfonic acids, Organic & Biomolecular Chemistry, DOI: 10.1039 / d1ob00552a) used sulfinic acid and olefins under blue light irradiation to synthesize compounds having the following β-ketosulfone structure; Yangfeng Li and Yizhou Li et al. used sodium sulfinate and olefins under iodine catalysis to synthesize vinyl sulfones (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-hydroxysulfone 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]

[0005] Traditional pharmacodynamic compound screening involves synthesizing individual small molecules with pharmacophores using organic synthesis methods. DNA-Encoded Chemical Library (DEL) technology, on the other hand, is an emerging small molecule screening technology based on combinatorial chemistry principles that allows for the rapid construction of ultra-large compound libraries. With the increasing application of technologies such as artificial intelligence and deep learning across various industries, the use of AI to accelerate new drug discovery is gaining increasing popularity. Because AI is often data-driven, using mathematical models to capture underlying patterns in data, its implementation in the biopharmaceutical field requires the rapid generation and accumulation of high-quality biological big data. DEL technology, with its vast chemical space spanning tens to hundreds of billions of chemical species, generates massive amounts of standardized experimental data, providing a crucial source for training reliable AI models. Deeply mining DEL data using AI algorithms, particularly by systematically analyzing the affinity of structurally similar molecules for related targets, increases the likelihood of discovering small molecules with unique structural properties, accelerating lead compound discovery and ultimately the entire drug discovery process. The novel DEL + AI model offers boundless opportunities and potential for future small molecule drug discovery.

[0006] However, after extensive literature searches, the inventors of this application have not found a method for rapidly constructing a large number of sulfinic acid structure compounds at one time, which cannot meet the DEL+AI model's need for screening effective compounds with sulfonyl groups. Summary of the Invention

[0007] In order to solve the need of DEL+AI mode screening of pharmacological compounds, this 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 said substituted R1 is halogen, alkyl or alkoxy;

[0008] Preferably, the substituted or unsubstituted alkyl group of R1 is a C2-C3 alkyl group; the substituted or unsubstituted aryl group of R1 is a six-membered aryl group; the heteroatom in the substituted or unsubstituted heterocyclic aromatic group of R1 is N, and the number of heteroatoms is 1; the substituent of R1 is fluorine, C1 alkyl or alkoxy.

[0009] 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 the proton bound to the sulfinate group corresponding to the base, 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 carbonates are more conducive to the purification of subsequent products because the carbonate group is converted into carbon dioxide and water during the reaction, which is more conducive to the requirements of pharmaceutical compounds.

[0010] 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 to 3:7. During the applicant's experiments, it was found that a mixture of DMSO and DMF with water performed significantly better than a mixture of MeCN, EtOH, THF, and 1,4-dioxane with water. When the ratio of water to DMSO is within the range of 3:97 to 3:7, and other reaction conditions remain the same, the conversion rate of the sulfinic acid structure compound gradually decreases.

[0011] The temperature is no higher than 80°C. During the experiment, the inventors of the present application found that, under the same other conditions, simply raising the reaction temperature to 80°C for one hour resulted in good DNA integrity and a conversion rate of nearly 70% for the On-DNA sulfinic acid structure compound. This conversion rate was significantly higher than the conversion rate of reacting at 60°C for one hour. Moreover, the conversion rate of the On-DNA sulfinic acid structure compound continued to gradually increase within the reaction range of room temperature to 80°C.

[0012] The light source has a wavelength of 365 to 535 nanometers. During experiments, the reaction was able to proceed even without light, achieving a yield of 35%. Conversion of the on-DNA sulfinic acid compound initially increased and then decreased when the light source wavelength was within the 365 to 535 nanometer range, so a wavelength of 365 to 405 nanometers is preferred. Irradiation with light effectively lowers the reaction temperature, resulting in milder reaction conditions. For example, without light irradiation, the on-DNA sulfinic acid compound conversion rate was 69% at 80°C for one hour. However, under the same other reaction conditions as above, irradiation with light at room temperature for 15 minutes achieved a conversion rate of 77%.

[0013] Preferably, dithiothreitol is added to the reaction, preferably at a final concentration of 0.1-1.0 mmol / L, where the final concentration is the concentration in the reaction system. Under the same conditions, the addition of DTT to the reaction system effectively increases the conversion rate of the On-DNA sulfinic acid structure compound by 20%.

[0014] 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 a compound with the same pharmacophore at a hundred or even a thousand times the order of magnitude of a small molecule organic chemistry. 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

[0015] Figure 1-19 : Liquid chromatography mass spectra of oligonucleic acid-sulfinic acid structure compounds 2a-2s;

[0016] Figure 20 : Liquid chromatography mass spectrum of oligonucleic acid-alkenyl sulfone structure compound 3m obtained in application example; DETAILED DESCRIPTION

[0017] DIEA: N,N-diisopropylethylamine;

[0018] DBU: abbreviation of bicyclic amidine, also known as bicyclic amidine, chemical name is 1,8-diazabicyclo (5,4,0)-7-undecene;

[0019] TEA: triethylamine

[0020] pH = 9.5 borate buffer;

[0021] DABCO: triethylenediamine;

[0022] DTT: dithiothreitol;

[0023] DMSO: dimethyl sulfoxide;

[0024] MeCN: acetonitrile;

[0025] DMF: N,N-dimethylformamide;

[0026] EtOH: ethanol;

[0027] 1,4-dioxane:1,4-dioxane;

[0028] THF: tetrahydrofuran;

[0029] 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 (a 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 the same method as application number 2024119940775 to prepare HP-Linker-NH2 and the structure of oligonucleic acid-thiol (phenol) compounds. Therefore, the structural confirmation diagrams of HP-NH2, HP-Linker-NHFmoc, HP-Linker-NH2, and oligonucleic acid-thiol (phenol) compounds 1a-1s are omitted.

[0030] 1. Synthesis of oligonucleic acid-linker-NH2 raw material (HP-Linker-NH2)

[0031] 1.1 Preparation of HP-Linker-NHFmoc from HP-NH2 (commercially available reagent) has the following reaction equation:

[0032]

[0033] Specifically, 100.0 nanomoles of HP-NH2 (commercially available) was dissolved in deionized water to prepare a 1.0 mmol / L solution (100.0 μL, 100.0 nanomoles, 1.0 equivalent). 40.0 equivalents of S1 (commercially available) in DMSO (concentration: 200.0 mmol / L), 250.0 equivalents of sodium tetraborate (Na2B4O7) buffer (concentration: 250.0 mmol / L) at pH 9.5, and 40.0 equivalents of 4-(4,6-dimethoxytriazin-2-yl)-4-methylmorpholine hydrochloride (DMT-MM) aqueous solution (concentration: 200.0 mmol / L) were mixed and thoroughly vortexed. This mixture was then added to the HP-NH2 solution, mixed thoroughly, 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, add anhydrous ethanol (3 times the total volume) and shake thoroughly. Chill the reaction mixture at -80°C for 2 hours. Centrifuge at 4000 rpm for 30 minutes, and discard the supernatant. Dissolve the remaining precipitate in deionized water to obtain a solution of HP-Linker-NHFmoc. Liquid chromatography-mass spectrometry analysis reveals a molecular weight of 5406 for HP-Linker-NHFmoc.

[0034] 1.2 Preparation of HP-Linker-NH2 from HP-Linker-NHFmoc:

[0035]

[0036] 100.0 nanomoles of HP-Linker-NHFmoc was dissolved in deionized water to prepare 1.0 mmol / L (100.0 microliters, 100.0 nanomoles, 1.0 equivalents), 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, 10% of the total volume of 5.0 mol / L sodium chloride solution was added to the reaction solution. Then, 3 times the total volume of anhydrous ethanol was added, and after shaking evenly, the reaction was placed in a refrigerator at -80°C for 2 hours. After that, centrifuged at 4000.0 rpm for half an hour, and the supernatant was discarded. The remaining precipitate was dissolved in deionized water to obtain a solution of oligonucleic acid-Linker-NH2 (HP-Linker-NH2). HP-Linker-NH2 was detected by liquid chromatography-mass spectrometry, and its molecular weight was 5184.

[0037] 2. Sources or synthesis methods of raw materials used to prepare oligonucleic acid-thiol (phenol) compounds:

[0038] This 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℃.

[0039] More specifically, S2j, S2l, and S2p were prepared by dissolving 0.3 g (1.3 eq.) of sodium nitrite (NaNO2) in 1.0 ml of ice-cold aqueous solution. The solution was then added dropwise at 0°C to a mixed solution of 3.3 mmol (1.0 eq.) of aminocarboxylic acid in 1.0 ml of aqueous solution and 0.6 ml of concentrated hydrochloric acid (HCl). The mixture was stirred at 5°C for 30 minutes. Subsequently, 1.04 g (1.3 eq.) of sodium sulfide nonahydrate (Na2S.9H2O) in 1.0 ml of cooled aqueous solution, 138.0 mg (0.16 eq.) of sulfur (S8), and 0.3 ml of a 10.0 mol / L aqueous sodium hydroxide (NaOH) solution were added in sequence. The mixture was stirred at room temperature for 2 hours and then acidified to pH 2 with hydrochloric acid. The resulting precipitate was collected by filtration, washed with water, dried under high vacuum, and carried to the next step without further purification.

[0040] 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 disulfide as a yellow solid.

[0041] 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] + .

[0042] 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. 1H 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] + .

[0043] 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] + .

[0044] 3. Preparation of oligonucleic acid-thiol (phenol) compounds

[0045] 3.1 Preparation of oligonucleic acid-thiol (phenol) compounds from HP-Linker-NH2

[0046] 3.1.1 Synthesis of oligonucleic acid-thiol (phenol) compound starting material (1a) from HP-Linker-NH2

[0047] 50.0 nmol of HP-Linker-NH2 was dissolved in deionized water to prepare 1.0 mmol / L (50.0 μl, 50.0 nmol, final concentration was 0.2 mmol / L). A DMSO solution of disulfide S2a (commercially available) (200.0 mmol / L, 50.0 μL, final concentration 40.0 mmol / L), EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 1,200.0 mmol / L, 50.0 μL, final concentration 240.0 mmol / L), HOAt (1-hydroxy-7-azobenzotriazole, 240.0 mmol / L, 50.0 μL, final concentration 48.0 mmol / L), and DIEA (N,N-diisopropylethylamine, 1,200.0 mmol / L, 50.0 μL, final concentration 240.0 mmol / L) was mixed and thoroughly vortexed. This mixture was then added to the HP-Linker-NH2 solution, mixed thoroughly, and reacted at 25°C for 1 hour. After the reaction is complete, 10% of the total volume of a 5.0 mol / L sodium chloride solution is added to the reaction solution. Then, 3 times the total volume of anhydrous ethanol is added. After vortexing, the reaction solution is placed in a -80°C freezer for 2 hours. The solution is then centrifuged at 4000.0 rpm for half an hour, and the supernatant is discarded. The remaining precipitate is treated with DTT to cleave disulfide bonds (patent: CN 114853822B), followed by precipitation with sodium chloride / ethanol and dissolution in deionized water to yield a solution of 1a with a molecular weight of 5272.6 and a yield of 76%.

[0048] The inventors of this application used the method for preparing raw material 1a to synthesize other oligonucleic acid-thiol (phenol) compounds (1b-g, 1j-1q).

[0049] 3.1.2 Synthesis of oligonucleic acid-thiol (phenol) compound raw materials (1h) from HP-Linker-NH2

[0050] Dissolve 50.0 nmol of HP-Linker-NH2 in deionized water to a final concentration of 1.0 mmol / L (50.0 μL, 50.0 nmol, 0.2 mmol / L). Combine a DMSO solution of sulfide S2h (200.0 mmol / L, 50.0 μL, 40.0 mmol / L), EDCI (1,200.0 mmol / L, 50.0 μL, 240.0 mmol / L), HOAt (240.0 mmol / L, 50.0 μL, 48.0 mmol / L), and DIEA (1,200.0 mmol / L, 50.0 μL, 240.0 mmol / L) and thoroughly mix on a vortex. Add this mixture to the HP-Linker-NH2 solution, mix thoroughly, and incubate at 25°C for 1 hour. After the reaction is complete, add 10% of the total volume of 5.0 mol / L sodium chloride solution to the reaction solution. Then, add three times the total volume of anhydrous ethanol. After vortexing, place the reaction solution in a -80°C freezer for 2 hours. Centrifuge at 4000 rpm for 30 minutes, and discard the supernatant. Dissolve the remaining precipitate in deionized water to obtain a 1-hour solution with a molecular weight of 5338.9 and a yield of >99%.

[0051] The inventor team of this application used the method for preparing raw material 1h to synthesize other oligonucleic acid-thiol (phenol) compounds (1i, 1r-1s).

[0052] 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.

[0053] Example 1-31 takes the following reaction as an example to optimize the reaction conditions 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.

[0054]

[0055]

[0056]

[0057] Examples 1-3 were conducted under laboratory fluorescent light without any additional light source. Comparing with Examples 1-4, we can see that although heating can convert the raw materials into the target products, light source can effectively improve the conversion rate.

[0058] Comparing Examples 5-8, we can see that, with other reaction conditions remaining unchanged, the conversion of the On-DNA sulfinic acid compound initially increases and then decreases when the light source wavelength is within the 365-535 nm range. Therefore, a wavelength of 365-405 nm is preferred for the reaction. Compared to conditions without light source heating, the light source significantly shortens the reaction time, increases the conversion rate, and makes the reaction conditions milder and more convenient.

[0059] Comparing Examples 6 and 9, we can see that when other reaction conditions remain unchanged, without the addition of 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 to a final concentration of 0.1-1.0 mmol / L.

[0060] 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.

[0061] 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. However, when NaHCO3 is removed, the 2m conversion rate drops sharply. Therefore, it is preferred to add NaHCO3 to a final concentration of 0.1-2.0 mmol / L.

[0062] 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 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.

[0063] 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 solvent is 3:97-3:7.

[0064] Comparing Examples 28-31, we can see that, with other reaction conditions remaining unchanged, when the final concentration of 1M is 0.05 mmol / L, the conversion rate of 2M is superior to that of the others. The inventors of this application discovered during experiments that, even within the range of 0.005 mmol / L to 0.2 mmol / L, to maintain the conversion rate unchanged, for example, to achieve a conversion rate of 91%, simply increasing the amount of other materials can be sufficient. However, for cost-effectiveness, a range of 0.01 to 0.2 mmol / L is preferred, and 0.02 to 0.2 mmol / L is more preferred.

[0065] Example 32 Synthesis of oligonucleic acid-sulfinic acid compound (2a) using oligonucleic acid-thiol (phenol) compound raw material (1a)

[0066]

[0067] To a 0.6 ml centrifuge tube were added an oligonucleic acid-thiol (phenol) compound (1a, 5.0 nmol, 1.0 equivalent, 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 of DMSO. 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 a 5.0 mol / L sodium chloride solution and 3 times the volume of anhydrous ethanol were added to the reaction solution. After vortexing, the solution was placed in a -80°C refrigerator for 2 hours, and then subjected to high-speed refrigerated centrifugation (4°C, 12,000 rpm, 15 minutes). The supernatant was discarded, and the remaining precipitate was the product, an oligonucleic acid-sulfinic acid structure compound (2a). The product was detected by liquid chromatography-mass spectrometry, and the molecular weight was 5326.6, and the conversion rate was >99%.

[0068] Examples 33-50

[0069] The difference from Example 32 is that oligonucleic acid-thiol (phenol) compounds (1b-1s) are used as raw materials to synthesize other representative oligonucleic acid-sulfinic acid structure compounds (2b-2s). The specific structures are as follows. The test results of 2a-2s are shown in Figure 1-19 .

[0070] The applicant would like to point out 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 is not exhaustive.

[0071] Application Examples

[0072]

[0073] In a 0.6 ml centrifuge tube, oligonucleic acid-sulfinic acid compound (2 m, 5.0 nmol, 1.0 equivalent, 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 vortexing and then shaken 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 shaking evenly, it was placed in a -80°C refrigerator and frozen for 2 hours, and then high-speed refrigerated centrifugation (4°C, 12,000 rpm, 15 minutes) was performed. The supernatant was discarded, and the remaining precipitate was the product oligonucleic acid-alkenyl sulfone structure compound (3 m, Figure 20 ).

[0074] 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 non-creative modifications to the present embodiment as needed, but as long as they are within the scope of the claims of the present application, they are protected by the patent law.

Claims

1. A method for preparing an On-DNA sulfinic acid structure compound, comprising the steps of preparing an On-DNA sulfinic acid structure compound from an On-DNA thiol or an On-DNA thiophenol, wherein 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 R 1 is a substituted or unsubstituted C2-C3 alkyl group, wherein R 1 is a substituted or unsubstituted aryl group, a six-membered aryl group, wherein 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; 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 solvent includes water and an organic solvent, and the organic solvent is dimethyl sulfoxide, N,N-dimethylformamide, acetonitrile, ethanol or tetrahydrofuran; The temperature is not higher than 80°C.

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

3. The method for preparing an On-DNA sulfinic acid structure compound according to claim 1, wherein: The ratio of water to organic solvent is 3:97-3:

7.

4. The method for preparing an On-DNA sulfinic acid structure compound according to claim 1, wherein: The reaction step of preparing the On-DNA sulfinic acid structure compound from On-DNA thiol or On-DNA thiophenol further uses light source irradiation, and the light source is 365 nanometers to 535 nanometers.

5. The method for preparing an On-DNA sulfinic acid structure compound according to claim 4, wherein: The light source is 365 to 405 nanometers.

6. The method for preparing an On-DNA sulfinic acid structure compound according to claim 1, wherein: Dithiothreitol was also added to the reaction.

7. The method for preparing an On-DNA sulfinic acid structure compound according to claim 6, wherein: The concentration of dithiothreitol is 0.1-1.0 mmol / L final concentration.

8. Using the method for preparing an On-DNA sulfinic acid structure compound according to any one of claims 1 to 7 for preparing a sulfinic acid structure compound.

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