Preparation method for on-DNA aromatic compound, and application of preparation method

Abstract

Provided are a preparation method for an On-DNA aromatic compound, and an application of the preparation method. The preparation method comprises the following steps: mixing an On-DNA aryl halide with an amino compound, a palladium catalyst, an alkali, a reducing agent and a solvent for reaction at 10-100°C to obtain the On-DNA aromatic compound. The provided preparation method can implement the preparation of On-DNA aromatic compounds within a wide temperature range, and can implement effective preparation of On-DNA aromatic compounds at room temperature and lower temperatures, thereby avoiding damage to DNA marker fragments.

Description

Preparation method and application of On-DNA aromatic compound

[0001] This application claims priority to Chinese invention patent application CN202311286574.5, filed on September 25, 2023, and incorporates the entire text of the patent application into this application. Technical Field

[0002] The invention belongs to the field of compound synthesis, and particularly relates to a preparation method and application of an On-DNA aromatic compound. Background Art

[0003] In new drug development, high-throughput screening of biological targets is one of the primary methods for rapidly identifying lead compounds. However, traditional high-throughput screening based on single molecules is time-consuming, requires significant equipment investment, and has a limited library size (millions of compounds). Building a compound library requires decades of accumulation, limiting the efficiency and potential for lead compound discovery. In 1992, Professors Sydney Brenner and Richard Lerner of the Scripps Research Institute in the United States first proposed the concept of a DNA-encoded library (DEL) (Reference: Proc. Natl. Acad. Sci., 1992, 89, 5381; Patent: US5573905). This method combines combinatorial chemistry and molecular biology techniques. At the molecular level, each organic small molecule is linked to a unique sequence of DNA (i.e., the organic small molecule reagent is DNA-tagged). The "combine-split" strategy of combinatorial chemistry is used to rapidly construct a large number of compound libraries through two or more cycles. Each compound in the library is composed of a different organic small molecule reagent residue and is marked by a corresponding unique base sequence of DNA. A very small amount of DNA-encoded compound library is then subjected to affinity screening with the target. Compound library molecules that do not form adsorption with the target are washed away, while molecules that do form adsorption with the target are further eluted. The library is then purified by DNA polymerase chain reaction (PCR). Reaction, abbreviated as PCR, can replicate and amplify the DNA labeled fragments in the compound library molecules that are adsorbed to the target until the amount of DNA obtained can be recognized by the DNA sequencer. By analyzing the relationship table between organic small molecules and each specific DNA base sequence, small molecules that can identify potentially active molecules are found. These small molecules are synthesized through traditional organic synthesis methods, and then their physiological activity against the target is tested and confirmed.

[0004] One of the greatest challenges in constructing DNA-encoded compound libraries is the need to efficiently synthesize chemically diverse small molecules on DNA. DNA must remain stable in a specific aqueous phase, pH, temperature, metal ion concentration, and inorganic salt concentration. Furthermore, the on-DNA reactions used to construct DNA-encoded compound libraries must also exhibit high yields. The reagent types, reaction types, and reaction conditions of the chemical reactions performed on DNA (referred to as on-DNA reactions) directly impact the richness and selectivity of DNA-encoded compound libraries. Consequently, developing DNA-compatible chemical reactions has become one of the most important research areas in DNA-encoded compound library technology, directly impacting the application and commercial value of DNA-encoded compound libraries.

[0005] In small molecule drugs, amine structural fragments are important structures that can participate in regulating drug solubility, pKa, and permeability. Therefore, as one of the most effective methods for constructing C-N bonds, the Buchwald-Hartwig reaction has been widely used in the synthesis of small molecule drugs. Currently, the publicly reported Buchwald-Hartwig catalyzed carbon-nitrogen bond coupling reaction conditions suitable for application in DNA-encoded compound libraries are mainly the following: Lu et al. from GSK and Shanghai Institute of Materia Medica first reported in 2017 the use of tButylXPhos-Buchwald precatalyst and CsOH to catalyze the Buchwald-Hartwig coupling reaction of On-DNA aryl iodides with aromatic primary amines in aqueous phase at 100°C (reference: Med. Chem. Commun., 2017, 8, 1614-1617); Simmons et al. from Baylor College of Medicine reported in 2020 the use of Pd-PEPPSI-iPentCl-pyr and CsOH to catalyze the Buchwald-Hartwig coupling reaction of On-DNA aryl chlorides, bromides and iodides with aromatic amines / secondary cyclic aliphatic amines in aqueous phase at 95°C (reference: Bioconjugate Chem., 2020, 31, 3, 770-780); Waring et al. from Newcastle University reported in 2021 that [(Cinnamyl)PdCl]2, cBRIDP, TPGS-750-M and potassium tert-butoxide catalyzed the Buchwald-Hartwig coupling reaction of On-DNA aryl chlorides, bromines and iodides with aromatic amines in aqueous phase at 80°C (reference: J.Org.Chem., 2021, 86(2 3), 17257-17264); Chheda et al. from Janssen reported in 2022 the use of BippyPhos, Pd(OAc)2 and potassium phosphate to catalyze the Buchwald-Hartwig coupling reaction of On-DNA aromatic chlorides, bromides and iodides with aromatic amines / aliphatic amines / amides / ureas in aqueous phase at 95°C (reference: Org. Lett., 2022, 24(18), 3401-3406).

[0006] However, the reaction temperatures for these on-DNA Buchwald-Hartwig coupling reactions are generally high, especially when the on-DNA aryl halide is brominated or chlorinated. These relatively high temperatures inevitably damage the labeled DNA fragments, thereby affecting the yield of DNA-encoded compound libraries. Lowering the reaction temperature to minimize damage to the labeled DNA fragments during the construction of DNA-encoded compound libraries has been a key research direction in the development of on-DNA chemistry.

[0007] Therefore, how to provide an On-DNA Buchwald-Hartwig carbon-nitrogen bond coupling reaction method that can be carried out at room temperature, is easy to operate, and is adaptable to large-scale multi-well plates, and then convert On-DNA aromatic halides and amino compounds into On-DNA aromatic amine compounds, thereby further enhancing the application value of DNA-encoded compound library technology, has become an urgent problem to be solved.

[0008] Summary of the Invention

[0009] To address the shortcomings of the prior art, the present invention provides a method for preparing on-DNA aromatic compounds and their application. The method provided by the present invention has good applicability and can be used to prepare on-DNA aromatic compounds over a wide temperature range. In particular, it can be effectively prepared at room temperature or even lower temperatures, avoiding damage to the labeled DNA fragments.

[0010] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0011] In one aspect, the present invention provides a method for preparing an On-DNA aromatic compound, the method comprising the following steps:

[0012] The On-DNA aromatic halide is reacted with an amino compound, a palladium catalyst, a base, a reducing agent, and a solvent at 10-100° C. to obtain the On-DNA aromatic compound;

[0013] The reaction formula is as follows:

[0014] Wherein DNA-Ar-X is On-DNA aryl halide, For amino compounds, Indicates a single bond or absence;

[0015] Ar is selected from any one of substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-10 membered heteroaryl, the number of the substituted groups is one or at least two, and the substituted groups are independently selected from deuterium, halogen, cyano, hydroxyl, amino, thiol, carboxyl, ester, nitro, aldehyde, C1-C20 alkyl, C1-C20 heteroalkyl, C2-C20 alkenyl, C2-C20 alkynyl, C6-C10 aryl, 5-10 membered heteroaryl, C any one of a 3-C12 cycloalkyl group or a 3-12 membered heterocyclic group; when the number of substituted groups is at least two, the substituted groups exist independently or any two substituted groups are linked by a covalent bond to form a C6-C10 aryl group, a 5-10 membered heteroaryl group, a C3-C12 cycloalkyl group or a 3-12 membered heterocyclic group which is unsubstituted or substituted by at least one substituent independently selected from deuterium, halogen, cyano, hydroxyl, amino, thiol, carboxyl, ester, nitro, aldehyde or oxo.

[0016] X is selected from Cl, Br or I.

[0017] R is selected from any one of substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-10 membered heteroaryl, the number of the substituted groups is one or at least two, and the substituted groups are independently selected from deuterium, halogen, cyano, hydroxyl, amino, nitro, sulfhydryl, carboxyl, ester, nitro, aldehyde, C1-C20 alkyl, C1-C20 heteroalkyl, C2-C20 alkenyl, C2-C20 alkynyl, C6-C10 aryl, 5-10 membered heteroaryl, C any one of a 3-C12 cycloalkyl group or a 3-12 membered heterocyclic group; when the number of substituted groups is at least two, the substituted groups exist independently or any two substituted groups are linked by a covalent bond to form a C6-C10 aryl group, a 5-10 membered heteroaryl group, a C3-C12 cycloalkyl group or a 3-12 membered heterocyclic group which is unsubstituted or substituted by at least one substituent independently selected from deuterium, halogen, cyano, hydroxyl, amino, nitro, thiol, carboxyl, ester, nitro, aldehyde or oxo.

[0018] When absent, R' is selected from any one of H, substituted or unsubstituted C1-C6 alkyl.

[0019] When it is a single bond, R and R' form any one of a substituted or unsubstituted C6-C10 aryl group and a substituted or unsubstituted 5-10 membered heteroaryl group with the N atom to which they are connected, the number of the substituted groups is one or at least two, and the substituted groups are independently selected from deuterium, halogen, cyano, hydroxyl, amino, nitro, thiol, carboxyl, ester, nitro, aldehyde, C1-C20 alkyl, C1-C20 heteroalkyl, C2-C20 alkenyl, C2-C20 alkynyl, C6-C10 aryl, 5-10 membered heteroaryl, any one of a C3-C12 cycloalkyl group or a 3-12 membered heterocyclic group; when the number of substituted groups is at least two, the substituted groups exist independently or any two substituted groups are linked by a covalent bond to form a C6-C10 aryl group, a 5-10 membered heteroaryl group, a C3-C12 cycloalkyl group or a 3-12 membered heterocyclic group which is unsubstituted or substituted by at least one substituent independently selected from deuterium, halogen, cyano, hydroxyl, amino, nitro, thiol, carboxyl, ester, nitro, aldehyde or oxo.

[0020] The catalyst is selected from any one or a combination of at least two of the following:

[0021] It should be noted that, in the above structural formula, Cy2 is a cyclohexyl group, TMS is a trimethylsilyl group, and i-Pr is an isopropyl group.

[0022] The above reaction uses a specific catalyst to achieve the effective preparation of On-DNA aromatic compounds at room temperature or lower temperatures, avoiding damage to the DNA labeled fragments.

[0023] Preferably, Ar is selected from the following substituted or unsubstituted groups:

[0024] wherein Y is selected from O, S or NR c , R c selected from H or C1-C6 alkyl, The end is connected to DNA or X.

[0025] Preferably, the group replacing Ar is selected from any one of C1-C20 alkyl, C1-C20 heteroalkyl, C2-C20 alkenyl, C2-C20 alkynyl, C6-C10 aryl, 5-10 membered heteroaryl, C3-C12 cycloalkyl or 3-12 membered heterocyclyl and the number is at least two, and any two substituted groups are linked by covalent bonds to form a C6-C10 aryl, 5-10 membered heteroaryl, C3-C12 cycloalkyl or 3-12 membered heterocyclyl that is unsubstituted or substituted by 1-4 identical or different substituents selected from deuterium, halogen, cyano, hydroxyl, amino, thiol, carboxyl, ester, nitro, aldehyde or oxo.

[0026] Preferably, Ar is selected from the following groups:

[0027]

[0028] The end is connected to X;

[0029] wherein R1 is selected from hydrogen, halogen, amino, nitro, cyano, hydroxyl, ester, thiol, C1-C12 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, 4-10 membered heterocyclyl, C6-C10 aryl, 5-10 membered heteroaryl, -C(O)C1-C12 alkyl, -C(O)C1-C6 alkoxy, -C(O)C3-C8 cycloalkyl, -C(O)4-10 membered heterocyclyl, -C(O)C6-C10 aryl, -C(O)5-10 membered heteroaryl, -NR a R b , -O-C6-C10 aryl, -O-5-10 membered heteroaryl, -O-C3-C6 cycloalkyl, -O-4-10 membered heterocyclyl, -C(O)NR a C1-C6 alkyl, -C(O)NR a C6-C10 aryl, -C(O)NR a 5-10 membered heteroaryl, -C(O)NR a C3-C6 cycloalkyl, -C(O)NR a 4-10 membered heterocyclic group, -NR a C(O)C1-C6 alkyl, -NR a C(O)C6-C10 aryl, -NR a C(O)5-10 membered heteroaryl, -NR a C(O)C3-C6 cycloalkyl, -NR a C(O)4-10 membered heterocyclyl; the C1-C12 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, 4-10 membered heterocyclyl, C6-C10 aryl, 5-10 membered heteroaryl, -C(O)C1-C12 alkyl, -C(O)C1-C6 alkoxy, -C(O)C3-C8 cycloalkyl, -C(O)4-10 membered heterocyclyl, -C(O)C6-C10 aryl, -C(O)5-10 membered heteroaryl, -O-C6-C10 aryl, -O-5-10 membered heteroaryl, -O-C3-C6 cycloalkyl, -O-4-10 membered heterocyclyl, -C(O)NR a C1-C6 alkyl, -C(O)NR a C6-C10 aryl, -C(O)NR a 5-10 membered heteroaryl, -C(O)NRa C3-C6 cycloalkyl, -C(O)NR a 4-10 membered heterocyclic group, -NR a C(O)C1-C6 alkyl, -NR a C(O)C6-C10 aryl, -NR a C(O)5-10 membered heteroaryl, -NR a C(O)C3-C6 cycloalkyl, -NR a The C(O)4-10 membered heterocyclyl is unsubstituted or substituted with at least one substituent independently selected from deuterium, halogen, cyano, hydroxy, amino, thiol, carboxyl, C1-C6 alkyl, C1-C6 alkoxy, ester, nitro, aldehyde or oxo.

[0030] R a 、R b R is independently selected from H, C1-C6 alkyl, C3-C8 cycloalkyl, 4-10 membered heterocyclyl, C6-C10 aryl, 5-10 membered heteroaryl; wherein the C1-C6 alkyl, C3-C8 cycloalkyl, 4-10 membered heterocyclyl, C6-C10 aryl, 5-10 membered heteroaryl is unsubstituted or independently substituted with one or at least two substituents selected from deuterium, halogen, cyano, hydroxyl, amino, thiol, carboxyl, C1-C6 alkyl, C1-C6 alkoxy, ester, nitro, aldehyde or oxo; or R a 、R b Together with the nitrogen atom to which they are attached, they form a 4-10 membered heterocyclic group, and the 4-10 membered heterocyclic group is unsubstituted or substituted by one or at least two substituents independently selected from deuterium, halogen, cyano, hydroxyl, amino, mercapto, carboxyl, C1-C6 alkyl, C1-C6 alkoxy, ester, nitro, aldehyde or oxo.

[0031] R2 is a group connected to the DNA portion, selected from -NH-, -C(=O)-, -C(=O)-O-, -NH-C(=O)-, -NH-C(=O)-CH2-, -NH-C(=O)-CH2-CH2-, -NH-C(=O)-NH-, -NH-S(=O)2-, -NH-S(=O)2-NH-, -NH-C(=O)-(CH2CH2O) n-CH2CH2C(=O)-NH-CH2-, any one of substituted or unsubstituted C1-C12 alkylene, substituted or unsubstituted C3-C12 cycloalkylene, and substituted or unsubstituted C6-C10 arylene; the substituted groups are independently selected from any one of deuterium, halogen, cyano, hydroxyl, amino, nitro, thiol, carboxyl, ester, nitro, aldehyde, C1-C20 alkyl, C1-C20 heteroalkyl, C2-C20 alkenyl, C2-C20 alkynyl, C6-C10 aryl, 5-10 membered heteroaryl, C3-C12 cycloalkyl, and 3-12 membered heterocyclyl.

[0032] Y is selected from O, S or NR c , R c Selected from H or C1-C6 alkyl.

[0033] Preferably, Ar is selected from the following groups:

[0034] Connect the X end.

[0035] Preferably, R is selected from any one of substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-6 membered heteroaryl, the number of the substituted groups is one or at least two, and the substituted groups are independently selected from deuterium, halogen, cyano, hydroxyl, amino, nitro, sulfhydryl, carboxyl, ester, nitro, aldehyde, C1-C10 alkyl, C1-C10 heteroalkyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, 5-6 membered heteroaryl, C3 -C7 cycloalkyl or any one of 3-7 membered heterocyclic groups; when the number of substituted groups is at least two, the substituted groups exist independently or any two substituted groups are linked by a covalent bond to form a C6-C10 aryl group, a 5-10 membered heteroaryl group, a C3-C12 cycloalkyl group or a 3-12 membered heterocyclic group which is unsubstituted or substituted with at least one substituent independently selected from deuterium, halogen, cyano, hydroxyl, amino, nitro, thiol, carboxyl, ester, nitro, aldehyde or oxo.

[0036] Preferably, R is selected from substituted or unsubstituted benzene, substituted or unsubstituted pyridine, substituted or unsubstituted imidazole, substituted or unsubstituted thiazole, substituted or unsubstituted pyrazole, substituted or unsubstituted thiophene, substituted or unsubstituted oxazole, substituted or unsubstituted furazan, substituted or unsubstituted pyrimidine, substituted or unsubstituted pyridazine, substituted or unsubstituted pyrazine, substituted or unsubstituted morpholine; the number of the substituted groups is one or at least two, and the substituted groups are independently selected from deuterium, halogen, cyano, hydroxyl, amino, nitro, thiol, carboxyl, ester, nitro, aldehyde, C1-C6 any one of alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, phenyl, 5-6 membered heteroaryl, C3-C6 cycloalkyl or 4-7 membered heterocyclyl; when the number of substituted groups is at least two, the substituted groups exist independently or any two substituted groups are linked by a covalent bond to form a C6-C10 aryl, 5-10 membered heteroaryl, C3-C7 cycloalkyl or 3-7 membered heterocyclyl which is unsubstituted or substituted by at least one substituent independently selected from deuterium, halogen, cyano, hydroxyl, amino, nitro, thiol, carboxyl, ester, nitro, aldehyde or oxo.

[0037] Preferably, the amino compound is a primary aromatic amine, R is selected from C6-C10 aryl, 5-10 membered heteroaryl, wherein the C6-C10 aryl, 5-10 membered heteroaryl is optionally replaced by one or more independently selected from halogen, hydroxyl, carboxyl, nitro, ester, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, 4-10 membered heterocyclyl, -C(O)C1-C6 alkyl, -C(O)NR a R b 、-NR a C(O)C1-C6 alkyl, -O-C6-C10 aryl substituents, and C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, 4-10 membered heterocyclyl, -C(O)C1-C6 alkyl, -C(O)NR a R b 、-NR a C(O)C1-C6 alkyl, -O-C6-C10 aryl is optionally substituted by one or more substituents independently selected from halogen, hydroxy, carboxyl, nitro, ester, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy; or

[0038] The amino compound It is a secondary amino compound, R is selected from C6-C10 aryl, 5-10 membered heteroaryl, wherein C6-C10 aryl, 5-10 membered heteroaryl is optionally replaced by one or more independently selected from halogen, hydroxyl, carboxyl, nitro, ester, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, 4-10 membered heterocyclyl, -C(O)C1-C6 alkyl, -C(O)NR a R b 、-NR a C(O)C1-C6 alkyl, -O-C6-C10 aryl substituents, and C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, 4-10 membered heterocyclyl, -C(O)C1-C6 alkyl, -C(O)NR a R b 、-NR a C (O) C1-C6 alkyl, -O-C6-C10 aryl is optionally substituted by one or more substituents independently selected from halogen, hydroxy, carboxyl, nitro, ester, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy; R' is selected from C1-C6 alkyl; or R and R' form a C6-C10 aryl, a 5-10 membered heteroaryl with the N atom to which they are attached, and the C6-C10 aryl, the 5-10 membered heteroaryl is optionally substituted by one or more substituents independently selected from halogen, hydroxy, carboxyl, nitro, ester, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, a 5-10 membered heteroaryl, C3-C6 cycloalkyl, a 4-10 membered heterocyclyl, -C (O) C1-C6 alkyl, -C (O) NR a R b 、-NR a C(O)C1-C6 alkyl, -O-C6-C10 aryl substituents, and C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, 4-10 membered heterocyclyl, -C(O)C1-C6 alkyl, -C(O)NR a R b 、-NR a C(O)C1-C6 alkyl, -O-C6-C10 aryl are optionally substituted by one or more substituents independently selected from halogen, hydroxy, carboxyl, nitro, ester, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy.

[0039] Preferably, the amino compound is a primary aromatic amine, R is selected from C6-C10 aryl, 5-10 membered heteroaryl, wherein the C6-C10 aryl, 5-10 membered heteroaryl is optionally replaced by one or more independently selected from halogen, hydroxyl, carboxyl, nitro, ester, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, 4-10 membered heterocyclyl, -C(O)C1-C6 alkyl, -C(O)NR a R b 、-NR a C(O)C1-C6 alkyl, -O-phenyl substituted, and as a substituent of C6-C10 aryl, 5-10 membered heteroaryl C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, 4-10 membered heterocyclyl, -C(O)C1-C6 alkyl, -C(O)NR a R b 、-NR a C(O)C1-C6 alkyl, -O-phenyl are optionally substituted by one or more substituents independently selected from halogen, hydroxy, carboxyl, nitro, ester, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy.

[0040] Preferably, the palladium catalyst is selected from [2-(dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [2-(dicyclohexylphosphine)-3-isopropoxy-2',4',6'-triisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [2-(dicyclohexylphosphine)-3-tert-butoxy-2',4',6'-triisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [2- (Dicyclohexylphosphine)-3-tert-butoxy-2',6'-diisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [2-(dicyclohexylphosphine)-3-tert-butoxy-6-methoxy-2',4',6'-triisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [2-(dicyclohexylphosphine)-3-tert-butoxy-6-methoxy-2',6'-diisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide or a combination of at least two thereof.

[0041] More preferably, the palladium catalyst is [2-(dicyclohexylphosphine)-3-tert-butoxy-6-methoxy-2',6'-diisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide.

[0042] The specific palladium catalyst can effectively realize the effective preparation of On-DNA aromatic compounds at room temperature or lower temperature, and can improve the raw material conversion rate.

[0043] Preferably, the solvent is an aqueous mixed solvent containing any one or at least two of dimethylacetamide, dimethylformamide, N-methylpyrrolidone, hexamethylphosphoric triamide, acetonitrile, dimethyl sulfoxide, methanol, ethanol, tert-butanol, isopropanol, tetrahydrofuran, 1,4-dioxane, an inorganic salt buffer solution, an organic acid buffer solution or an organic base buffer solution.

[0044] Preferably, the solvent is a mixed solvent of hexamethylphosphoric triamide and water.

[0045] Preferably, the water content in the reaction liquid is not less than 20%.

[0046] The above-mentioned specific solvent can effectively improve the conversion rate of raw materials.

[0047] Preferably, the base includes any one or a combination of at least two of sodium borate, potassium borate, potassium phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, sodium acetate, sodium fluoride, potassium fluoride, cesium fluoride, lithium hydroxide, sodium hydroxide, potassium hydroxide, cesium hydroxide, lithium carbonate, sodium carbonate, potassium carbonate, cesium carbonate, sodium bicarbonate or potassium bicarbonate, preferably any one or a combination of at least two of lithium hydroxide, sodium hydroxide, potassium hydroxide or cesium hydroxide.

[0048] Preferably, the reducing agent comprises sodium ascorbyl palmitate.

[0049] Preferably, the molar ratio of the On-DNA aryl halide to the amino compound, palladium catalyst, base, and reducing agent is 1:(10-1000):(1-100):(10-1000):(20-500), that is, the equivalents of the amino compound, palladium catalyst, base, and reducing agent relative to the On-DNA aryl halide are 10-1000, 1-100, 10-1000, and 20-500, respectively; preferably 1:(250-1000):(1-100):(50-1000):(100-250).

[0050] Preferably, the molar ratio of the On-DNA aryl halide to the palladium catalyst is 1:(1-100); for example, 1:1, 1:2, 1:5, 1:10, 1:15, 1:20, 1:30, 1:40, 1:50, 1:60, 1:70, 1:80, 1:90, 1:100.

[0051] Preferably, the molar ratio of the On-DNA aryl halide to the palladium catalyst is 1:(2-20).

[0052] Preferably, the molar ratio of the On-DNA aryl halide to the amine compound is 1:(10-1000); for example, 1:10, 1:20, 1:50, 1:100, 1:200, 1:250, 1:300, 1:400, 1:500, 1:600, 1:700, 1:800, 1:900, 1:1000.

[0053] Preferably, the molar ratio of the On-DNA aryl halide to the amino compound is 1:(20-1000).

[0054] Preferably, the molar ratio of the On-DNA aryl halide to the amino compound is 1:(50-1000).

[0055] Preferably, the molar ratio of the On-DNA aryl halide to the amino compound is 1:(50-500).

[0056] Preferably, the molar ratio of the On-DNA aryl halide to the amino compound is 1:(100-500).

[0057] Preferably, the molar ratio of the On-DNA aryl halide to the base is 1:(10-1000); for example, 1:10, 1:20, 1:50, 1:100, 1:200, 1:250, 1:300, 1:400, 1:500, 1:600, 1:700, 1:800, 1:900, 1:1000.

[0058] Preferably, the molar ratio of the On-DNA aryl halide to the base is 1:(50-1000).

[0059] Preferably, the molar ratio of the On-DNA aryl halide to the base is 1:(100-500).

[0060] Preferably, the molar ratio of the On-DNA aryl halide to the reducing agent is 1:(20-500); for example, 1:20, 1:50, 1:100, 1:200, 1:250, 1:300, 1:400, 1:500.

[0061] Preferably, the molar ratio of the On-DNA aryl halide to the reducing agent is 1:(100-250).

[0062] Preferably, the reaction temperature is 10-80°C, for example 10°C, 20°C, 30°C, 40°C, 50°C, 60°C, 70°C or 80°C, but is not limited to the values ​​listed above. Other unlisted values ​​within the above numerical range are also applicable, preferably 10-60°C, more preferably 20-50°C, further preferably 20-40°C, further preferably 20-30°C, further preferably 25°C or room temperature.

[0063] Preferably, the reaction time is 0.5-12 h, for example, 0.5 h, 1 h, 2 h, 3 h, 4 h, 5 h, 6 h, 7 h, 8 h, 9 h, 10 h, 11 h or 12 h, but is not limited to the above values. Other values ​​not listed in the above numerical range are also applicable, preferably 1-6 h, more preferably 1-2 h.

[0064] On the other hand, the present invention also provides the use of the above-mentioned method for preparing the On-DNA aromatic compound in constructing a DNA-encoded compound library.

[0065] Compared with the prior art, the present invention has the following beneficial effects:

[0066] The present invention provides a method for preparing On-DNA aromatic compounds with excellent applicability. The method is applicable to various On-DNA aromatic halides, amino compounds, bases, and solvents, and has a wide range of reaction conditions. The method of the present invention uses a specific palladium catalyst to achieve the preparation of On-DNA aromatic compounds through the Buchwald-Hartwing reaction. In particular, the method can achieve effective preparation of On-DNA aromatic compounds under lower temperature conditions (even room temperature or lower temperature conditions) than the existing Buchwald-Hartwing reaction, avoiding damage to DNA marker fragments. The method is particularly suitable for preparing DNA-encoded compound libraries. DETAILED DESCRIPTION

[0067] In order to further illustrate the technical means and effects adopted by the present invention, the technical solutions of the present invention are further described below in conjunction with the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.

[0068] The term "alkyl" is used to represent a straight-chain or branched saturated hydrocarbon group. For example, a C1-C3 alkyl group refers to a saturated hydrocarbon group containing 1 to 3 carbon atoms, a C1-C4 alkyl group refers to a saturated hydrocarbon group containing 1 to 4 carbon atoms, and a C1-C6 alkyl group refers to a saturated hydrocarbon group containing 1 to 6 carbon atoms, including straight-chain or branched alkyl groups, and so on. No further details are given.

[0069] The term "cycloalkyl" refers to a cyclized alkyl group based on a non-aromatic carbocyclic ring. Cycloalkyl groups can include monocyclic, bicyclic, or polycyclic ring systems. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. C3-C8 cycloalkyl groups refer to cycloalkyl groups containing 3 to 8 carbon atoms, and so on.

[0070] The term "alkenyl" is used to refer to a straight-chain or branched unsaturated hydrocarbon group containing one or at least two carbon-carbon double bonds. For example, C2-C3 alkenyl refers to an unsaturated hydrocarbon group containing 1 to 3 carbon atoms and one or two carbon-carbon double bonds, C2-C4 alkenyl refers to an unsaturated hydrocarbon group containing 1 to 4 carbon atoms and one or two carbon-carbon double bonds, and C1-C6 alkenyl refers to an unsaturated hydrocarbon group containing 1 to 6 carbon atoms and one or at least two carbon-carbon double bonds, including straight-chain or branched hydrocarbon groups, and so on. No further details are given.

[0071] The term "alkynyl" is used to refer to a straight-chain or branched unsaturated hydrocarbon group containing one or at least two carbon-carbon triple bonds. For example, a C2-C3 alkynyl group refers to an unsaturated hydrocarbon group containing 1 to 3 carbon atoms and one carbon-carbon triple bond, a C2-C4 alkynyl group refers to an unsaturated hydrocarbon group containing 1 to 4 carbon atoms and one or two carbon-carbon triple bonds, and a C1-C6 alkynyl group refers to an unsaturated hydrocarbon group containing 1 to 6 carbon atoms and one or at least two carbon-carbon triple bonds, including straight-chain or branched hydrocarbon groups, and so on. No further details are given.

[0072] The term "alkoxy" is used to represent a structure formed by a straight-chain or branched hydrocarbon group connected to an oxygen atom. For example, a C1-C3 alkoxy group refers to a substituent containing 1 to 3 carbon atoms and connected to an oxygen atom, a C2-C4 alkoxy group refers to a substituent containing 1 to 4 carbon atoms and connected to an oxygen atom, and a C1-C6 alkoxy group refers to a substituent containing 1 to 6 carbon atoms and connected to an oxygen atom, wherein the hydrocarbon portion includes a straight-chain or branched hydrocarbon group. Examples of alkoxy groups include methoxy, ethoxy, n-propyloxy, etc., and so on, which are not repeated here.

[0073] The term "heteroalkyl" is used to indicate a straight or branched saturated hydrocarbon group in which at least one carbon atom is replaced by a heteroatom selected from O, N and S atoms.

[0074] The term "heterocycloalkyl" or "heterocyclyl" refers to a non-aromatic heterocycloalkyl group in which one or more ring carbon atoms are replaced by heteroatoms selected from O, N and S atoms. "Heterocyclyl" may include monocyclic, bicyclic or polycyclic ring systems.

[0075] "Aryl" refers to an aromatic carbocyclic group, including monocyclic, bicyclic, tricyclic or polycyclic aromatic hydrocarbons, such as phenyl, naphthyl, anthracenyl, phenanthrenyl, etc. Aryl is preferably a monocyclic, bicyclic or tricyclic ring system having 6 to 14 or 6 to 12 ring members. Preferably, at least one ring in the system is aromatic and each ring in the system contains 3 to 7 ring members. "Substituted aryl" refers to an aryl group in which at least one hydrogen atom on the phenyl ring is replaced by a non-hydrogen moiety. Preferred aryl groups include phenyl, biphenyl, indanyl, 1-naphthyl, 2-naphthyl and tetrahydronaphthyl.

[0076] The term "heteroaromatic ring" or "heteroaryl" means a 3-12 membered aromatic monocyclic, bicyclic or polycyclic heterocyclic ring that is fully unsaturated, partially unsaturated, and contains carbon atoms and one or more heteroatoms independently selected from N, O and S. The heteroaryl herein is preferably a 3-12 membered heteroaryl. The heteroaryl herein preferably has one, two or three heteroatoms independently selected from N, O and S. Preferably, the heteroaryl herein is any heterocyclic ring as defined above fused to a benzene ring. The nitrogen and sulfur heteroatoms may be optionally oxidized. The nitrogen atom may be substituted or unsubstituted (i.e., N or NR, where R is H or, if defined, another substituent). The heterocyclic ring may be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure. The heterocyclic groups described herein may be substituted on carbon or nitrogen atoms if the resulting compound is stable. The nitrogen in the heterocyclic ring may be optionally quaternized. Preferably, when the total number of S and O atoms in the heterocycle exceeds 1, then these heteroatoms are not adjacent to each other.Examples of aromatic hetero groups include, but are not limited to, acridinyl, azetidinyl, azcinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiopyranyl, benzoxazolyl, benzoxazolinyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, quinolinyl, decahydroquinolinyl, dihydrofuro[2,3-b]tetrahydrofuranyl, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, imidazopyridinyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, iso Benzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isothiazolopyridinyl, isoxazolyl, isoxazolopyridinyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolopyridinyl, oxazolidinyl, perylene, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl , 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolopyridinyl, pyrazolyl, pyridazinyl, pyridooxazolyl, pyridoimidazolyl, pyridothiazolyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2-pyrrolidinonyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinazolinyl, quinuclidinyl, tetrazolyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl , thianthrenyl, thiazolyl, thiopyranyl, thiazolopyridinyl, thiopyranothiazolyl, thiopyranoxazolyl, thiopyranoimidazolyl, thiopyranyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl and xanthene, quinolinyl, isoquinolinyl, phthalazinyl, quinazolinyl, indolyl, 1H-indazolyl, benzimidazolyl, 1,2,3,4-tetrahydroquinolinyl, 1,2,3,4-tetrahydroisoquinolinyl, 5,6,7,8-tetrahydro-quinolinyl, 2,3-dihydro-benzofuranyl, 1,2,3,4-tetrahydro-quinazolinyl and 1,2,3,4-tetrahydro-quinazolinyl. The term "heteroaryl" may also include biaryl structures formed by the above-defined "aryl" and a monocyclic "heteroaryl", such as but not limited to "-phenylbipyridyl-", "-phenylbipyrimidyl-", "-pyridylbiphenyl-", "-pyridylbipyrimidyl-", and "-pyrimidylbiphenyl-".

[0077] The term "halogen" includes fluorine, chlorine, bromine and iodine.

[0078] The term "substituted" refers to the replacement of one or more hydrogen atoms on a specific group with a specific substituent. The specific substituent is the substituent described above or the substituent appearing in the examples. Unless otherwise specified, a substituted group may have a substituent selected from the specific group at any substitutable position of the group, and the substituent may be the same or different at each position. It will be understood by those skilled in the art that the combinations of substituents contemplated by the present invention are those that provide stable or chemically feasible compounds. Typical substitutions include, but are not limited to, one or more (e.g., 1 to 3) of the following groups: hydrogen, deuterium, halogen, hydroxyl (-OH), nitrile, nitro, oxo (e.g., =O, where two H atoms on a carbon atom are replaced by it to form a carbonyl group), alkyl (C1-6 alkyl), cycloalkyl (e.g., C3-6 cycloalkyl), alkenyl, alkynyl, heterocycloalkyl (e.g., 4-10 membered heterocycloalkyl), aryl, heteroaryl (e.g., 5-6 membered heteroaryl), alkoxy (e.g., C1-6 alkoxy, such as methoxy, ethoxy), C1-6 haloalkyl (e.g., single or multiple halogen substitutions), halogen substituted, the latter such as trifluoromethyl), C1-6 haloalkoxy (for example, monohalogen substituted or polyhalogen substituted, the latter such as trifluoromethoxy), -OC(=O)NRiiRiii, ORi, SRi, S(=O)Rv, S(=O)2Rv, P(=O)2Rv, S(=O)2ORv, P(=O)2ORv, NRiiRiii, NRiiS(=O)2Rv, NRiiP(=O)Rv, S(=O)2NRiiRiii, P(=O)2NRiiRiii, C(=O)O Riv, C(=O)Ri, C(=O)NRiiRiii, OC(=O)Ri, OC(=O)NRiiRiii, NRiiC(=O)ORv, NRivC(=O)NRiiRiii, NRivS(=O)2NRiiRiii, NRivP(=O)2NRiiRiii, NRiiC(=O)Ri, or NRiiP(=O)2Rv, wherein Ri can independently represent hydrogen, deuterium, alkyl, cycloalkyl, alkenyl, alkynyl, heterocyclic or aromatic ring, and Rii, Riii and R IV can independently represent hydrogen, deuterium, alkyl (e.g., C1-C6 alkyl), cycloalkyl (e.g., C3-C6 cycloalkyl), heterocycloalkyl, or aryl, or Rii and Riii together with the nitrogen atom to which they are attached can form a heterocycloalkyl (e.g., a 4-10 membered heterocycloalkyl), which is optionally substituted with 1-3 substituents selected from halogen, -OH, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, or C1-C6 haloalkoxy; Rv can independently represent hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, heterocycle, or aromatic ring. The above-mentioned typical substituents, such as alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or aromatic ring, can be optionally substituted.The substituents include, but are not limited to, halogen, hydroxyl, cyano, carboxyl (-COOH), C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3-12 membered heterocyclic group, aryl, heteroaryl, C1-C8 aldehyde, C2-C10 acyl, C2-C10 ester, amino, C1-C6 alkoxy, C1-C10 sulfonyl, and C1-C6 urea, etc.

[0079] In the following examples, the DNA-NH2 structure used is shown below (Chengdu Pioneer, 05082023):

[0080] Wherein, A is adenine, T is thymine, C is cytosine, and D is guanine; however, this application is not limited to a specific DNA structure, and other structural DNAs can also implement the preparation method of this application, such as the structure shown below:

[0081] (Chengdu Pioneer, 08302022).

[0082] The remaining reagents used are as follows:

[0083] DMA: dimethylacetamide.

[0084] EDCI: 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride.

[0085] s-NHS: N-hydroxysuccinimide sulfonic acid sodium salt.

[0086] DMSO: dimethyl sulfoxide.

[0087] HMPA: Hexamethylphosphoric triamide.

[0088] NaOH: sodium hydroxide.

[0089] Sodium borate buffer: borate buffer solution.

[0090] Palladium catalyst OA1 (BrettPhos Pd G6 TES) was purchased from Pepper Reagents, Catalog No. 601818; palladium catalyst OA2 (EPhos Pd G6 TES) was purchased from Pepper Reagents, Catalog No. 601823. GPhos Pd G3 TES: [2-(dicyclohexylphosphino)-3-tert-butoxy-6-methoxy-2',6'-diisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide (i.e., OA6) was purchased from Pepper Reagents, Catalog No. 602204.

[0091] The preparation method of catalyst OA3 is as follows:

[0092] Step 1 Preparation of compound 3-1

[0093] Flask A, equipped with a polytetrafluoroethylene stopper and a stirring bar, was dried using a heat gun. The flask was vacuumed and backfilled with nitrogen three times. m-Fluoroanisole (25.6 mmol) and anhydrous THF (tetrahydrofuran 85 mL) were added to the flask and the temperature of flask A was lowered to -78°C. 14.9 mL of a 2.0 M n-butyllithium (n-Bli) n-hexane solution was then slowly added dropwise to flask A over 10 minutes. The reactants in flask A were allowed to react at -78°C for 1 hour. Simultaneously, flask B was vacuumed and backfilled with nitrogen three times at room temperature. After adding elemental iodine (30 mmol) and anhydrous THF (tetrahydrofuran 50 mL), flask B was then cooled to -78°C. After flask A reacted for 1 hour, the solution in flask B was added dropwise to flask A. The whole process took more than 10 minutes. After the addition was completed, flask A was opened to the air and a saturated aqueous solution of Na2S2O3 (100 mL) was added to flask A. The mixture was warmed to room temperature and the organic matter was extracted using a separatory funnel and ether. Anhydrous magnesium sulfate was added to dry the organic solution, and the compound 3-1 was obtained by rotary evaporation.

[0094] Step 2 Preparation of compound 3-3

[0095] To a dry two-necked flask C, add magnesium metal (23.3 mmol, 20% powder, 80% shavings) and a condenser, then seal with a stopper. Evacuate and backfill with nitrogen three times. Add THF (tetrahydrofuran, 19 mL) and compound 3-2 (2-bromo-1,3,5-triisopropylbenzene, 10.1 mmol) to flask C. Place the entire flask in an oil bath preheated to 70°C and heat. Add 1,2-dibromoethane dropwise to flask C using a syringe. The reaction mixture in flask C is allowed to react at 70°C, monitoring the reaction progress using GC until the 2-bromo-1,3,5-triisopropylbenzene is completely consumed. Slowly add 3-1 (3-fluoro-2-iodo-anisole, 10.5 mmol) obtained in step 1 dropwise to flask C using a syringe. Monitor the reaction by GC until the reaction of the starting material 3-fluoro-2-iodo-anisole is complete. Cool flask C to room temperature. At this time, 50mL flask D was dried, vacuumed and backfilled with nitrogen three times after sealing, and elemental iodine (10.1mmol, 2.57g, 1.00 equivalent) and anhydrous THF (tetrahydrofuran 10mL) were added to flask D. The iodine solution in D was slowly added dropwise to flask C until the reaction mixture in flask C remained a lasting lavender. The addition was stopped. At this time, flask A was opened to the air, and its contents were transferred to a separatory funnel. A saturated aqueous solution of Na2S2O3 (50mL), brine (25mL) and ethyl acetate (50mL) were added. After separation of the organic layer, the organic phase was washed once with brine (25mL) and the organic solution was dried over anhydrous magnesium sulfate. The crude product was rotary evaporation, washed with methanol, and dried to give compound 3-3 after vacuum filtration.

[0096] Step 3 Preparation of compound 3-4

[0097] Use a heat gun to dry the flask with a polytetrafluoroethylene stirrer. Add compound 3-3 (2-iodo-2',4',6'-triisopropyl-3-methoxy-1,1'-biphenyl, 7.82 mmol) obtained in step 2 to the flask. Seal the flask with a rubber stopper, then drain the flask and backfill with nitrogen (repeat the vacuum / backfill process three times). Use a syringe to add anhydrous dichloromethane (50 mL) to the flask, place the flask in an ice-water bath and cool it to 0°C. Add BBr3 (boron tribromide 21.9 mmol) dropwise to the flask using a syringe. Then remove the flask from the ice-water bath and warm it to room temperature to continue the reaction for 1 hour. The flask was then placed in an ice-water bath and cooled to 0°C. The flask was opened and CH2Cl2 (25 mL of dichloromethane) and MeOH (10 mL of methanol) were carefully and slowly added to the flask. The mixture was then warmed to room temperature again, and saturated aqueous NaHCO3 solution (100 mL) was slowly and carefully added to the flask. The aqueous and organic phases of the mixture in the flask were separated, and the aqueous phase was extracted twice with dichloromethane (2 × 50 mL). The combined organic phases were dried over anhydrous MgSO4, filtered, and evaporated under reduced pressure to obtain compound 3-4.

[0098] Step 4 Preparation of Compound 3-5

[0099] A 300 mL flask A equipped with a Teflon stir bar was dried using a heat gun. Compound 3-4 (2-iodo-2',4',6'-triisopropylbiphenyl-3-ol, 7.82 mmol) was added to flask A and sealed. Flask A was then evacuated and backfilled with nitrogen (the evacuation / backfilling process was repeated three times). Anhydrous dichloromethane (58 mL) was added to the flask using a syringe, and flask A was cooled to -78°C. After drying, reaction tube B (Fisherbrand, 13 x 100 mm, product number 1495935C) was sealed with a screw cap (Fisherbrand, 13-425, C4015-66). The cap of reaction tube B was pierced with a needle attached to a Schlenk line. A balloon was connected to the other end of the Schlenk line. Vacuum was then applied to reaction tube B and the attached balloon, and the reaction tube B was cooled to -78°C. Then, another rubber tube with a needle was used. The rubber tube was connected to the isobutylene cylinder. The needle pierced the sealing cap of reaction tube B, and then isobutylene (23 mL, 117 mmol) was introduced into reaction tube B. The isobutylene in reaction tube B was quickly introduced into flask A through a catheter. The septum of bottle A was removed, and pure trifluoromethanesulfonic acid (30 drops of TfOH) was added dropwise with a glass pipette, and the bottle was resealed with a septum. The reaction process was monitored by thin layer chromatography until the phenol was completely consumed. Additional TfOH (10 drops) was added after 1.5 h, and additional isobutylene (10 mL) was added after 7.5 h. Flask A was removed from the cold water bath, and the septum was pierced with a needle to allow the isobutylene gas generated during the heating process to be discharged. The reaction was quenched with triethylamine (100 drops). The crude product mixture was concentrated under reduced pressure with the help of a rotary evaporator. Column chromatography (SiO2) was performed using two column volumes of hexane and hexane:ethyl acetate (5:1 v / v) as the eluent. All fractions containing the desired product were combined and concentrated under reduced pressure on a rotary evaporator to afford compound 3-5.

[0100] Step 5 Preparation of Compound 3-6

[0101] A dried round-bottom flask was sealed with a rubber septum. The septum of the reaction apparatus was penetrated by a needle connected to a Schlenk line using a rubber hose. A needle connected to a balloon was also inserted through the septum. The balloon and flask were evacuated and backfilled with argon (the evacuation / backfilling process was repeated three times; note: after this cycle, the balloon was inflated with argon). Anhydrous tetrahydrofuran (2.5 mL) was added via syringe, and the reaction flask was cooled to -78°C using a dry ice / acetone bath. 1.4 mL of a 1.7 M t-BuLi solution in pentane was added dropwise via syringe to the solution. The reaction mixture was stirred at -78°C for 1 hour. The reaction mixture became heterogeneous. Dicyclohexylphosphine chloride (CIPCy2, 1.2 mmol) was added dropwise via syringe to the solution. The cooling bath was removed, and the flask was allowed to warm to room temperature. The solution was stirred while heating for 3 hours. The crude reaction mixture was then opened, diluted with ethyl acetate (5 mL), and transferred to a separatory funnel. The organic layer was washed with saturated NH4Cl (2×15 mL), dried over MgSO4, filtered, and concentrated using a rotary evaporator to obtain a solid, which was then crystallized using MeOH (80 mL) to obtain compound 3-6.

[0102] Step 6 Preparation of palladium catalyst OA3

[0103] To an oven-dried round-bottom flask (containing a Teflon-coated magnetic stir bar) were added compound 3-6 (71 mg) obtained in Step 5 and ethyl 2-(trimethylsilyl)-4-bromobenzoate (77 mg). The flask was sealed with a rubber septum. A Schlenk line was connected using a rubber hose. The septum was pierced with a needle. The test tube or flask was evacuated and backfilled with nitrogen (the evacuation / backfilling process was repeated three times). Pentane (6 mL) was added via syringe until ligand L3 was completely dissolved. The septum was removed, and (COD)Pd(CH2TMS)2 (51 mg, 0.13 mmol, CAS No. 225931-80-6) was quickly added. The septum was immediately replaced, and the reaction mixture was stirred at room temperature for 16 h. The precipitate was collected by vacuum filtration, and the resulting filter cake was washed with additional pentane (3 × 5 mL). The resulting solid was dried under high vacuum for ≥2 h to yield palladium catalyst OA3.

[0104] 1H NMR (400MHz, CDCl3): δ7.52(d,J=8.0Hz,2H),7.23(m,3H),7.09(s,2H),7.04(d,J=8.5Hz,1H),6. 22(d,J=7.4Hz,1H),4.34(t,J=8.2Hz,2H),3.09(sept,J=6.9Hz,1H),2.95(q,J=11.9Hz,2H),2.5 3(sept,J=6.7Hz,2H),1.85–1.73(m,6H),1.72–1.60(m,6H),1.66(s,9H),1.62(d,J=6.9Hz,6H), 1.38(d,J=6.9Hz,6H),1.17(s,6H),1.08(t,J=8.3Hz,2H),0.91(d+m,J=6.7Hz,8H),0.05(s,9H).

[0105] 13 C{1H}NMR (101MHz, CDCl3): δ168.0,159.0,157.1,150.6,150.0,146.8,138.3,138.2,131.1,126.7,126.1,126.0,125.5,125.0, 123.3,123.0,114.0,80.4,62.6,35.8,35.5,34.4,31.7,29.6,29.4,29.17,27.8,27.7,27.6,26.1,25.7,25.0,24.9,17.5,-1.3.

[0106] 31 P{1H}NMR (162MHz, CDCl3): δ37.1.S19.

[0107] HRMS(ESI)calc.C 49 H 74 O3PPdSi + [M–Br] + :875.4174.Found:875.4177.

[0108] IR(Diamond-ATR,neat,cm -1 ):2929,1707,1573,1260,1160,837,797,754.

[0109] The preparation route of palladium catalyst OA4 can refer to the method of palladium catalyst OA3, and only the reaction substrate 3-2 of step 2 needs to be adaptively adjusted to OA4 characterization data are as follows:

[0110] 1 H NMR(400MHz,CDCl3):δ7.82(t,J=7.7Hz,1H),7.56(d,J=8.0Hz,2H),7.33–7.21(m,5H),7.13–7.06(m,1H),6.27(dd,J=7.6,2.4Hz,1H),4.36(t,J=8.2Hz,2H),3.00(q,J=12.3Hz,2H),2.56(sept,J=6.7Hz,2H),1.93–1.63(m,8H),1.69(s,9H),1.63(d,J=6.7Hz,6H),1.39–1.14(m,10H),1.10(t,J=8.3Hz,2H),0.99–0.82(m,2H),0.94(d,J=6.6Hz,6H),0.08(s,9H)。

[0111] 13 C{ 1 H}NMR(101MHz,CDCl3):δ167.9,159.2,159.1,150.6,149.6,149.4,145.3,145.3,138.3,138.2,134.1,131.3,126.9,126.6,125.9,125.8,122.9,122.6,114.3,114.3,80.6,62.7,35.8,35.6,31.6,29.6,29.4,29.2,29.2,27.8,27.7,27.7,27.6,26.1,25.7,24.9,22.5,17.5,14.2,-1.3。

[0112] 31 P{ 1 H}NMR(162MHz,CDCl3):δ37.5。

[0113] HRMS(ESI)calc.C 46 H 68 O3PPdSi + [M–Br] + :833.3705.Found:833.3711。

[0114] IR(Diamond-ATR,neat,cm -1 ):2918,1700,1573,1274,1258,1163,1009,838,756。

[0115] The preparation route of palladium catalyst OA5 can also refer to the method of palladium catalyst OA3, and the adaptability of 3-0 in step 1 needs to be adjusted to The characterization data are as follows:

[0116] 1 H NMR (400MHz, CDCl3): δ7.50(d,J=8.1Hz,2H),7.25(d,J=6.5Hz,2H),7.10–7.02(m,3H),6.73(d, J=9.1Hz,1H),4.40–4.28(m,2H),3.33(s,3H),3.08(sept,J=6.7Hz,1H),3.01–2.87(m,2H),2.5 0(sept,J=6.8Hz,2H),1.86–1.64(m,12H),1.63(s,9H),1.61(d,J=6.9Hz,6H),1.37(d,J=6.9Hz ,6H),1.17(d,J=7.6Hz,6H),1.11–1.04(m,2H),0.95(m,2H),0.82(d,J=6.6Hz,6H),0.05(s,9H).

[0117] 13 C{ 1 H}NMR (101MHz, CDCl3): δ168.1,157.5,152.3,151.8,151.8,151.2,151.1,147.5, 138.7,138.6,138.6,138.5,126.8,126.5,126.3,125.3,124.7,124.4,116.5,116.4,115.4,115.4,112.6,79.6,62.6,54.5,36.3,36.0,34.5,31.5,31.5,29.7,29.4,29.3,29.3,27.8,27.8,27.7,27.7,26.1,25.7,25.0,24.7,17.5,-1.3.

[0118] 31 P{ 1 H} NMR (162 MHz, CDCl3): δ 36.8.

[0119] HRMS(ESI)calc.C 50 H 76 O4PPdSi + [M–Br] + :905.4280.Found:905.4285.

[0120] IR(neat,cm-1 ):2932,1710,1574,1261,1161,838,825,756.

[0121] Example 1: Synthesis of On-DNA Aryl Halides

[0122] Taking p-bromobenzoic acid as an example reaction raw material, the reaction formula is as follows:

[0123] Dissolve 20 nmol DNA-NH2 in 20 μL of 250 mM, pH 9.5 borate buffer to a 1 mM concentration. Mix p-bromobenzoic acid (1 μmol, 50 equivalents, 400 mM DMA solution) with EDCI (1 μmol, 50 equivalents, 200 mM DMSO solution) and s-NHS condensation activator (0.6 μmol, 30 equivalents, 200 mM DMSO:H2O=2:1 mixed solution) for 15 minutes. Add the mixture to the DNA-NH2 raw material reaction solution, mix well, and incubate at 20°C overnight.

[0124] Standard post-processing for ethanol precipitation: Add 10% of the total volume of 5 M sodium chloride solution to the solution, followed by 3 times the total volume of chilled anhydrous ethanol. After vortexing, freeze the reaction at -80°C for 1 hour. Centrifuge at 12,000 rpm for 30 minutes and discard the supernatant. Dissolve the remaining precipitate in deionized water and ultrafilter twice to obtain the On-DNA aryl halide solution. Concentrate to dryness and use directly in the next reaction.

[0125] In the following examples, all On-DNA aryl halides used can be synthesized by referring to the above method.

[0126] Example 2: Synthesis of On-DNA Aromatic Amine Compounds

[0127] A series of On-DNA aryl halides were synthesized using the method of Example 1, and then reacted with aniline to obtain different On-DNA aromatic amine compounds. The exemplary reaction formula is as follows:

[0128] The On-DNA aryl halide synthesized in Example 1 was dissolved in ultrapure water to prepare a 1 mM concentration solution (10 μL, 10 nmol). Aniline (5 μmol, 500 equivalents, 12.5 μL, 400 mM HMPA solution), NaOH (2.5 μmol, 250 equivalents, 2.5 μL, 1 M aqueous solution), sodium ascorbyl palmitate (1 μmol, 100 equivalents, 5 μL, 200 mM aqueous solution), and GPhos Pd G3 TES (20 nmol, 2 equivalents, 2 μL, 10 mM HMPA solution) were added in sequence and mixed well. The mixture was sonicated for 2 minutes and reacted at 25°C for 2 hours.

[0129] Palladium removal: After the reaction is completed, add sodium diethyldithiocarbamate solution (5 μL, 1 μmol, 50 equivalents (relative to palladium metal), 200 mM aqueous solution) to the reaction vessel, centrifuge, let the solution settle to the bottom, vortex evenly, centrifuge again, and react at 50°C in a PCR instrument for 30 minutes (lid temperature: 105°C). After the reaction is completed, a large amount of precipitate is produced. Centrifuge at a centrifugal force of 4000G for 15 minutes, and transfer the supernatant to a new reaction tube for ethanol precipitation.

[0130] Ethanol precipitation: Add 10% of the total volume of 5M sodium chloride solution to the tube, and then continue to add 3 times the total volume of chilled anhydrous ethanol. After shaking evenly, place the reaction at -80°C and freeze for 1 hour. Then, centrifuge at 4000 rpm at 4°C for half an hour, remove the supernatant, wash the precipitate with deionized water, and vacuum freeze-dry at -40°C. Detect the OD by microplate reader to confirm the recovery rate, and send it to LCMS for detection to confirm the conversion rate of each small molecule.

[0131] The conversion rates of reaction products and raw materials (based on On-DNA aryl halides) are as follows:

[0132] Example of conversion results of On-DNA aryl chloride coupling reaction with aniline

[0133] Characterization data of some compounds in the above table are as follows:

[0134] Theoretical LCMS value: 5132.2; measured LCMS value: 5131.8.

[0135] Theoretical LCMS value: 5183.2; measured LCMS value: 5183.0.

[0136] Theoretical LCMS value: 5199.2; measured LCMS value: 5201.0.

[0137] The results showed that the actual products obtained were consistent with the theoretical expected products.

[0138] Example conversion results of the coupling reaction of On-DNA aryl bromide and aniline (the value under the compound structure is the conversion rate):

[0139] The characterization data of some of the above compounds are as follows:

[0140] Theoretical LCMS value: 5174.3; measured LCMS value: 5174.2.

[0141] Theoretical LCMS value: 5150.2; measured LCMS value: 5149.9.

[0142] Theoretical LCMS value: 5139.3; measured LCMS value: 5138.9.

[0143] The results showed that the actual products obtained were consistent with the theoretical expected products.

[0144] Example of conversion results of On-DNA aryl iodide coupling reaction with aniline

[0145] The characterization data of some of the above compounds are as follows:

[0146] Theoretical LCMS value: 5160.3; measured LCMS value: 5160.0.

[0147] Theoretical LCMS value: 5132.3; measured LCMS value: 5131.8.

[0148] Theoretical LCMS value: 5316.9; measured LCMS value: 5316.4.

[0149] The results showed that the actual products obtained were consistent with the theoretical expected products.

[0150] Example 3: Synthesis of On-DNA Aromatic Amine Compounds

[0151] Immobilized On-DNA aryl halides (with the same structure except for the halogen atoms) were synthesized using the method of Example 1, and then reacted with different amino compounds to obtain different On-DNA aromatic amine compounds. The exemplary reaction formulas are as follows:

[0152] The On-DNA aryl halide synthesized in Example 1 was dissolved in ultrapure water to prepare a 1 mM concentration solution (10 μL, 10 nmol). Aromatic amine compounds (5 μmol, 500 equivalents, 12.5 μL, 400 mM HMPA solution), NaOH (2.5 μmol, 250 equivalents, 2.5 μL, 1 M aqueous solution), sodium ascorbyl palmitate (2.5 μmol, 250 equivalents, 2.5 μL, 1 M aqueous solution), and GPhos Pd G3 TES (100 nmol, 10 equivalents, 2 μL, 50 mM HMPA solution) were added in sequence and mixed well. The mixture was sonicated for 2 minutes and reacted at 25°C for 2 hours.

[0153] Palladium removal: After the reaction is completed, add sodium diethyldithiocarbamate solution (5 μL, 5 μmol, 50 equivalents (relative to palladium metal), 1 M aqueous solution) to the reaction vessel, centrifuge, let the solution settle to the bottom, vortex evenly, centrifuge again, and react at 50°C in a PCR instrument for 30 minutes (cover temperature: 105°C). After the reaction is completed, a large amount of precipitate is produced. Centrifuge at a centrifugal force of 4000G for 15 minutes, and transfer the supernatant to a new reaction tube for ethanol precipitation.

[0154] Ethanol precipitation: Add 10% of the total volume of 5M sodium chloride solution to the tube, and then continue to add 3 times the total volume of chilled anhydrous ethanol. After shaking evenly, place the reaction at -80°C and freeze for 1 hour. Then, centrifuge at 4000 rpm at 4°C for half an hour, remove the supernatant, wash the precipitate with deionized water, and vacuum freeze-dry at -40°C. Detect the OD by microplate reader to confirm the recovery rate, and send it to LCMS for detection to confirm the conversion rate of each small molecule.

[0155] The reaction products and raw material conversion rates are as follows:

[0156] Exemplary conversion results of On-DNA coupling reactions of p-halobenzamide with different aromatic amines

[0157] Some other results are as follows (the values ​​under the compound structures are conversion rates, Br represents the conversion rate of Br substitutes, Cl represents the conversion rate of Cl substitutes, and I represents the conversion rate of I substitutes):

[0158] The characterization data of some of the above compounds are as follows:

[0159] Theoretical LCMS value: 5238.9; measured LCMS value: 5238.0.

[0160] Theoretical LCMS value: 5256.8; measured LCMS value: 5255.7.

[0161] Theoretical LCMS value: 5197.8; measured LCMS value: 5196.8.

[0162] The results showed that the actual products obtained were consistent with the theoretical expected products.

[0163] From the above results, it can be found that the preparation method provided by the present invention has good universality for a variety of amino compounds and On-DNA aromatic halides, and effectively realizes the preparation of On-DNA aromatic compounds with various structures.

[0164] Example 4: Study on the effects of different palladium catalyst types and dosages on the reaction

[0165] Using aniline and p-bromobenzamide DNA as raw materials, the effects of different palladium catalyst types and dosages on the reaction were studied (based on Example 2, the reducing agent vitamin C sodium equivalent was 250), and the results are as follows:

[0166] The results showed that OA1-OA6 could synthesize On-DNA aromatic compounds at 25℃.

[0167] Example 5: Study on the effects of different solvents on the reaction

[0168] Using aniline and p-bromobenzamide DNA as raw materials, the effects of different solvents on the reaction were studied (based on Example 2, the reducing agent vitamin C sodium equivalent was 250), and the results are as follows:

[0169] The results showed that various solvents could achieve the synthesis of On-DNA aromatic compounds at 25°C.

[0170] Example 6: Study on the effects of different base types and equivalents on the reaction

[0171] Using aniline and p-bromobenzamide DNA as raw materials, the effects of different base types and equivalents on the reaction were studied (based on Example 2, the reducing agent vitamin C sodium equivalent was 250), and the results are as follows:

[0172] The results showed that a variety of bases in a large equivalent range (especially 50-1000 equivalents, preferably 100-500 equivalents) can achieve the synthesis of On-DNA aromatic compounds at 25°C, and hydroxides perform better.

[0173] Example 7: Study on the Effect of Different Amine Compound Equivalents on the Reaction

[0174] Using aniline and p-bromobenzamide DNA as raw materials, the effect of different amine compound equivalents on the reaction was studied (based on Example 2, the reducing agent vitamin C sodium equivalent was 250), and the results are as follows:

[0175] The results show that amine compounds can achieve the synthesis of On-DNA aromatic compounds at 25°C within a large equivalent range (20-1000, preferably 100-1000, more preferably 100-500).

[0176] Example 8: Study on the effect of different reaction times on the reaction

[0177] Using aniline and p-bromobenzamide DNA as raw materials, the effects of different reaction times on the reaction were studied (based on Example 2, the reducing agent vitamin C sodium equivalent was 250), and the results are as follows:

[0178] The results show that the method of the present application can quickly realize the synthesis of On-DNA aromatic compounds at 25°C and can control the reaction time within 1-2h, which not only shortens the reaction time but also ensures a high conversion rate.

[0179] Example 9: Study on the influence of different reaction temperatures on the reaction

[0180] Using aniline and p-bromobenzamide DNA as raw materials, the effects of different reaction temperatures on the reaction were studied (based on Example 2, the reducing agent vitamin C sodium equivalent was 250), and the results are as follows:

[0181] The results show that the method of the present application can realize the synthesis of On-DNA aromatic compounds in a large temperature range, especially at a lower temperature, and can ensure the conversion rate of the reaction at room temperature (or 25°C).

[0182] The applicant states that while the above-described embodiments illustrate the preparation methods and applications of the On-DNA aromatic compounds, the present invention is not limited to these embodiments. This does not necessarily mean that the present invention must rely on these embodiments for implementation. Those skilled in the art will appreciate that any improvements to the present invention, equivalent substitutions for raw materials in the products of the present invention, additions of auxiliary ingredients, and selection of specific methods, etc., fall within the scope of protection and disclosure of the present invention.

[0183] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the scope of protection of the present invention.

[0184] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any appropriate manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A method for preparing an On-DNA aromatic compound, characterized in that: The preparation method comprises the following steps: The On-DNA aromatic halide is mixed with an amino compound, a palladium catalyst, a base, a reducing agent, and a solvent for reaction at 10-100° C. to obtain the On-DNA aromatic compound; The reaction formula is as follows: Wherein DNA-Ar-X is On-DNA aryl halide, It is an amino compound. Indicates a single bond or absence; Ar is selected from any one of substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-10 membered heteroaryl, the number of the substituted groups is one or at least two, and the substituted groups are independently selected from deuterium, halogen, cyano, hydroxyl, amino, thiol, carboxyl, ester, nitro, aldehyde, C1-C20 alkyl, C1-C20 heteroalkyl, C2-C20 alkenyl, C2-C20 alkynyl, C6-C10 aryl, 5-10 membered heteroaryl, C any one of 3-C12 cycloalkyl or 3-12 membered heterocyclic group; when the number of substituted groups is at least two, the substituted groups exist independently or any two substituted groups are linked by covalent bonds to form a C6-C10 aryl, 5-10 membered heteroaryl, C3-C12 cycloalkyl or 3-12 membered heterocyclic group which is unsubstituted or substituted by at least one substituent independently selected from deuterium, halogen, cyano, hydroxyl, amino, thiol, carboxyl, ester, nitro, aldehyde or oxo; X is selected from Cl, Br or I; R is selected from any one of substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-10 membered heteroaryl, the number of the substituted groups is one or at least two, and the substituted groups are independently selected from deuterium, halogen, cyano, hydroxyl, amino, nitro, thiol, carboxyl, ester, nitro, aldehyde, C1-C20 alkyl, C1-C20 heteroalkyl, C2-C20 alkenyl, C2-C20 alkynyl, C6-C10 aryl, 5-10 membered heteroaryl, C any one of 3-C12 cycloalkyl or 3-12 membered heterocyclic group; when the number of substituted groups is at least two, the substituted groups exist independently or any two substituted groups are linked by covalent bonds to form a C6-C10 aryl, 5-10 membered heteroaryl, C3-C12 cycloalkyl or 3-12 membered heterocyclic group which is unsubstituted or substituted by at least one substituent independently selected from deuterium, halogen, cyano, hydroxyl, amino, nitro, thiol, carboxyl, ester, nitro, aldehyde or oxo; When absent, R' is selected from any one of H, substituted or unsubstituted C1-C6 alkyl; When it is a single bond, R and R' form with the N atom to which they are connected any one of a substituted or unsubstituted C6-C10 aryl group, a substituted or unsubstituted 5-10 membered heteroaryl group, the number of the substituted groups is one or at least two, and the substituted groups are independently selected from deuterium, halogen, cyano, hydroxyl, amino, nitro, thiol, carboxyl, ester, nitro, aldehyde, C1-C20 alkyl, C1-C20 heteroalkyl, C2-C20 alkenyl, C2-C20 alkynyl, C6-C10 aryl, 5-10 membered heteroaryl, -10-membered heteroaryl, C3-C12 cycloalkyl or 3-12-membered heterocyclic group; when the number of substituted groups is at least two, the substituted groups exist independently or any two substituted groups are linked by covalent bonds to form a C6-C10 aryl, 5-10-membered heteroaryl, C3-C12 cycloalkyl or 3-12-membered heterocyclic group that is unsubstituted or substituted by at least one substituent independently selected from deuterium, halogen, cyano, hydroxyl, amino, nitro, thiol, carboxyl, ester, nitro, aldehyde or oxo; The catalyst is selected from [2-(dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [2-(dicyclohexylphosphine)-3-isopropoxy-2',4',6'-triisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [2-(dicyclohexylphosphine)-3-tert-butoxy-2',4',6'-triisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [2-(dicyclohexylphosphine)-3-tert-butoxy-2',4',6'-triisopropyl-1,1'-biphenyl](4 -((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [2-(dicyclohexylphosphine)-3-tert-butoxy-6-methoxy-2',4',6'-triisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [2-(dicyclohexylphosphine)-3-tert-butoxy-6-methoxy-2',6'-diisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [2-(dicyclohexylphosphine)-3-tert-butoxy-6-methoxy-2',6'-diisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, methanesulfonate [2-(dicyclohexylphosphine)-3-tert-butoxy-6-methoxy-2',6'-diisopropyl-1,1'-biphenyl](2'-amino-1,1'-biphenyl-2-yl)palladium, or a combination of at least two thereof.

2. The method for preparing an On-DNA aromatic compound according to claim 1, characterized in that: The Ar is selected from the following substituted or unsubstituted groups: Wherein Y is selected from O, S or NR c , R c Selected from H or C1-C6 alkyl, The end is connected to DNA or X; Preferably, the group replacing Ar is selected from any one of C1-C20 alkyl, C1-C20 heteroalkyl, C2-C20 alkenyl, C2-C20 alkynyl, C6-C10 aryl, 5-10 membered heteroaryl, C3-C12 cycloalkyl or 3-12 membered heterocyclyl and the number is at least two, and any two substituted groups are connected by covalent bonds to form a C6-C10 aryl, 5-10 membered heteroaryl, C3-C12 cycloalkyl or 3-12 membered heterocyclyl which is unsubstituted or substituted with 1-4 identical or different substituents selected from deuterium, halogen, cyano, hydroxyl, amino, thiol, carboxyl, ester, nitro, aldehyde or oxo.

3. The method for preparing an On-DNA aromatic compound according to claim 1 or 2, characterized in that: The Ar is selected from the following groups: wherein R1 is selected from hydrogen, halogen, amino, nitro, cyano, hydroxyl, thiol, ester, C1-C12 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, 4-10 membered heterocyclyl, C6-C10 aryl, 5-10 membered heteroaryl, -C(O)C1-C12 alkyl, -C(O)C1-C6 alkoxy, -C(O)C3-C8 cycloalkyl, -C(O)4-10 membered heterocyclyl, -C(O)C6-C10 aryl, -C(O)5-10 membered heteroaryl, -NR a R b , -O-C6-C10 aryl, -O-5-10 membered heteroaryl, -O-C3-C6 cycloalkyl, -O-4-10 membered heterocyclyl, -C(O)NR a C1-C6 alkyl, -C(O)NR a C6-C10 aryl, -C(O)NR a 5-10 membered heteroaryl, -C(O)NR a C3-C6 cycloalkyl, -C(O)NR a 4-10 membered heterocyclic group, -NR a C(O)C1-C6 alkyl, -NR a C(O)C6-C10 aryl, -NR a C(O)5-10 membered heteroaryl, -NR a C(O)C3-C6 cycloalkyl, -NR a C(O)4-10 membered heterocyclic group; the C1-C12 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, 4-10 membered heterocyclic group, C6-C10 aryl, 5-10 membered heteroaryl, -C(O)C1-C12 alkyl, -C(O)C1-C6 alkoxy, -C(O)C3-C8 Cycloalkyl, -C(O)4-10 membered heterocyclyl, -C(O)C6-C10 aryl, -C(O)5-10 membered heteroaryl, -O-C6-C10 aryl, -O-5-10 membered heteroaryl, -O-C3-C6 cycloalkyl, -O-4-10 membered heterocyclyl, -C(O)NR a C1-C6 alkyl, -C(O)NR a C6-C10 aryl, -C(O)NR a 5-10 membered heteroaryl, -C(O)NR a C3-C6 cycloalkyl, -C(O)NR a 4-10 membered heterocyclic group, -NR a C(O)C1-C6 alkyl, -NR a C(O)C6-C10 aryl, -NR a C(O)5-10 membered heteroaryl, -NR a C(O)C3-C6 cycloalkyl, -NR a C(O)4-10 membered heterocyclyl is unsubstituted or substituted by at least one substituent independently selected from deuterium, halogen, cyano, hydroxyl, amino, thiol, carboxyl, C1-C6 alkyl, C1-C6 alkoxy, ester, nitro, aldehyde or oxo; R a , R b R is independently selected from H, C1-C6 alkyl, C3-C8 cycloalkyl, 4-10 membered heterocyclyl, C6-C10 aryl, 5-10 membered heteroaryl; wherein the C1-C6 alkyl, C3-C8 cycloalkyl, 4-10 membered heterocyclyl, C6-C10 aryl, 5-10 membered heteroaryl is unsubstituted or independently substituted with one or at least two substituents selected from deuterium, halogen, cyano, hydroxyl, amino, thiol, carboxyl, C1-C6 alkyl, C1-C6 alkoxy, ester, nitro, aldehyde or oxo; or R a , R b Together with the nitrogen atom to which they are attached, they form a 4-10 membered heterocyclic group, and the 4-10 membered heterocyclic group is unsubstituted or substituted by one or at least two substituents independently selected from deuterium, halogen, cyano, hydroxyl, amino, mercapto, carboxyl, C1-C6 alkyl, C1-C6 alkoxy, ester, nitro, aldehyde or oxo; R2 is a group connected to the DNA portion, selected from -NH-, -C(=O)-, -C(=O)-O-, -NH-C(=O)-, -NH-C(=O)-CH2-, -NH-C(=O)-CH2-CH2-, -NH-C(=O)-NH-, -NH-S(=O)2-, -NH-S(=O)2-NH-, -NH-C(=O)-(CH2CH2O) n -CH2CH2C(=O)-NH-CH2-, any one of substituted or unsubstituted C1-C12 alkylene, substituted or unsubstituted C3-C12 cycloalkylene, and substituted or unsubstituted C6-C10 arylene; the substituted groups are independently selected from any one of deuterium, halogen, cyano, hydroxyl, amino, nitro, thiol, carboxyl, ester, nitro, aldehyde, C1-C20 alkyl, C1-C20 heteroalkyl, C2-C20 alkenyl, C2-C20 alkynyl, C6-C10 aryl, 5-10 membered heteroaryl, C3-C12 cycloalkyl, or 3-12 membered heterocyclyl; Y is selected from O, S or NR c , R c Selected from H or C1-C6 alkyl; The end is connected to X; Preferably, Ar is selected from the following groups: Connect the X end.

4. The method for preparing an On-DNA aromatic compound according to any one of claims 1 to 3, characterized in that: R is selected from any one of substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-6 membered heteroaryl, the number of the substituted groups is one or at least two, and the substituted groups are independently selected from deuterium, halogen, cyano, hydroxyl, amino, nitro, thiol, carboxyl, ester, nitro, aldehyde, C1-C10 alkyl, C1-C10 heteroalkyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, 5-6 membered heteroaryl, C3-C any one of 7-membered cycloalkyl or 3-7-membered heterocyclic group; when the number of substituted groups is at least two, the substituted groups exist independently or any two substituted groups are linked by covalent bonds to form a C6-C10 aryl, 5-10-membered heteroaryl, C3-C12 cycloalkyl or 3-12-membered heterocyclic group which is unsubstituted or substituted by at least one substituent independently selected from deuterium, halogen, cyano, hydroxyl, amino, nitro, thiol, carboxyl, ester, nitro, aldehyde or oxo; Preferably, R is selected from substituted or unsubstituted benzene, substituted or unsubstituted pyridine, substituted or unsubstituted imidazole, substituted or unsubstituted thiazole, substituted or unsubstituted pyrazole, substituted or unsubstituted thiophene, substituted or unsubstituted oxazole, substituted or unsubstituted furazan, substituted or unsubstituted pyrimidine, substituted or unsubstituted pyridazine, substituted or unsubstituted pyrazine, substituted or unsubstituted morpholine; the number of the substituted groups is one or at least two, and the substituted groups are independently selected from deuterium, halogen, cyano, hydroxyl, amino, nitro, sulfhydryl, carboxyl, ester, nitro, aldehyde , C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, phenyl, 5-6 membered heteroaryl, C3-C6 cycloalkyl or 4-7 membered heterocyclyl; when the number of substituted groups is at least two, the substituted groups exist independently or any two substituted groups are connected by covalent bonds to form C6-C10 aryl, 5-10 membered heteroaryl, C3-C7 cycloalkyl or 4-7 membered heterocyclyl which is unsubstituted or substituted by at least one substituent independently selected from deuterium, halogen, cyano, hydroxyl, amino, nitro, thiol, carboxyl, ester, nitro, aldehyde or oxo. Cycloalkyl or 3-7 membered heterocyclic group; Preferably, the amino compound is a primary aromatic amine, R is selected from C6-C10 aryl, 5-10 membered heteroaryl, wherein C6-C10 aryl, 5-10 membered heteroaryl is optionally replaced by one or more independently selected from halogen, hydroxyl, carboxyl, nitro, ester, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, 4-10 membered heterocyclyl, -C(O)C1-C6 alkyl, -C(O)NR a R b 、-NR a C(O)C1-C6 alkyl, -O-C6-C10 aryl substituents, C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, 4-10 membered heterocyclyl, -C(O)C1-C6 alkyl, -C(O)NR a R b 、-NR a C(O)C1-C6 alkyl, -O-C6-C10 aryl is optionally substituted by one or more substituents independently selected from halogen, hydroxy, carboxyl, nitro, ester, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy; or The amino compound is a secondary amino compound, R is selected from C6-C10 aryl, 5-10 membered heteroaryl, wherein C6-C10 aryl, 5-10 membered heteroaryl is optionally replaced by one or more independently selected from halogen, hydroxyl, carboxyl, nitro, ester, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, 4-10 membered heterocyclyl, -C(O)C1-C6 alkyl, -C(O)NR a R b 、-NR a C(O)C1-C6 alkyl, -O-C6-C10 aryl substituents, C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, 4-10 membered heterocyclyl, -C(O)C1-C6 alkyl, -C(O)NR a R b 、-NR a C(O)C1-C6 alkyl, -O-C6-C10 aryl is optionally substituted by one or more substituents independently selected from halogen, hydroxyl, carboxyl, nitro, ester, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy; R' is selected from C1-C6 alkyl; or R and R' form a C6-C10 aryl, a 5-10 membered heteroaryl with the N atom to which they are attached, and the C6-C10 aryl, the 5-10 membered heteroaryl is optionally substituted by one or more substituents independently selected from halogen, hydroxyl, carboxyl, nitro, ester, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, a 5-10 membered heteroaryl, C3-C6 cycloalkyl, a 4-10 membered heterocyclyl, -C(O)C1-C6 alkyl, -C(O)NR a R b 、-NR a C(O)C1-C6 alkyl, -O-C6-C10 aryl substituents, C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, 4-10 membered heterocyclyl, -C(O)C1-C6 alkyl, -C(O)NR a R b 、-NR a C(O)C1-C6 alkyl, -O-C6-C10 aryl are optionally substituted by one or more substituents independently selected from halogen, hydroxy, carboxyl, nitro, ester, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy.

5. The method for preparing an On-DNA aromatic compound according to any one of claims 1 to 4, characterized in that: The palladium catalyst is selected from [2-(dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [2-(dicyclohexylphosphine)-3-isopropoxy-2',4',6'-triisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [(2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [2-(dicyclohexylphosphine)-3-tert-butoxy-2',4',6'-triisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [2-(dicyclohexylphosphine)-3-tert-butoxy-2',6'-diisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [(2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [2-(dicyclohexylphosphine)-3-tert-butoxy-6-methoxy-2',4',6'-triisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide, [2-(dicyclohexylphosphine)-3-tert-butoxy-6-methoxy-2',4',6'-triisopropyl-1,1'-biphenyl] -biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide or a combination of at least two thereof. Further preferably, the palladium catalyst is [2-(dicyclohexylphosphine)-3-tert-butoxy-6-methoxy-2',6'-diisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide.

6. The method for preparing an On-DNA aromatic compound according to any one of claims 1 to 5, characterized in that: The solvent is an aqueous mixed solvent containing any one or at least two of dimethylacetamide, dimethylformamide, N-methylpyrrolidone, hexamethylphosphoric triamide, acetonitrile, dimethyl sulfoxide, methanol ethanol, tert-butyl alcohol, isopropanol, tetrahydrofuran, 1,4-dioxane, an inorganic salt buffer solution, an organic acid buffer solution or an organic base buffer solution; Preferably, the solvent is a mixed solvent of hexamethylphosphoric triamide and water; Preferably, the water content in the reaction liquid of the reaction is not less than 20%.

7. The method for preparing an On-DNA aromatic compound according to any one of claims 1 to 6, characterized in that: The base includes any one or a combination of at least two of sodium borate, potassium borate, potassium phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, sodium acetate, sodium fluoride, potassium fluoride, cesium fluoride, lithium hydroxide, sodium hydroxide, potassium hydroxide, cesium hydroxide, lithium carbonate, sodium carbonate, potassium carbonate, cesium carbonate, sodium bicarbonate or potassium bicarbonate, preferably any one or a combination of at least two of lithium hydroxide, sodium hydroxide, potassium hydroxide or cesium hydroxide; Preferably, the reducing agent comprises sodium vitamin C.

8. The method for preparing an On-DNA aromatic compound according to any one of claims 1 to 7, characterized in that: The molar ratio of the On-DNA aryl halide to the palladium catalyst is 1:(1-100), preferably 1:(2-20); and / or The molar ratio of the On-DNA aryl halide to the amine compound is 1:(10-1000), preferably 1:(20-1000), further preferably 1:(50-1000), further preferably 1:(50-500), more preferably 1:(100-500); and / or The molar ratio of the On-DNA aryl halide to the base is 1:(10-1000), preferably 1:(50-1000), and further preferably 1:(100-500); and / or The molar ratio of the On-DNA aryl halide to the reducing agent is 1:(20-500), preferably 1:(100-250).

9. The method for preparing an On-DNA aromatic compound according to any one of claims 1 to 8, characterized in that: The reaction temperature is 10-80°C, preferably 10-60°C, more preferably 20-50°C, more preferably 20-40°C, more preferably 20-30°C, more preferably 25°C; and / or The reaction time is 0.5-12 h, preferably 1-6 h, and more preferably 1-2 h.

10. Use of the method for preparing an On-DNA aromatic compound according to any one of claims 1 to 9 in constructing a DNA-encoded compound library.

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