A method for removing a benzyl group from a DNA-encoded compound

Abstract

This invention relates to a method for removing benzyl groups from DNA-encoded compounds. The method uses on-DNA compounds containing benzyl-protected amino and / or hydroxyl groups as raw materials, reacting them in the presence of a palladium catalyst and a hydrogen source to obtain on-DNA compounds containing amino and / or hydroxyl groups. The method for removing benzyl groups from DNA-encoded compounds provided by this invention can be carried out in a mixed aqueous phase of organic solvent / aqueous phase, with simple post-processing, mild conditions, and can obtain a highly diverse library of DNA-encoded compounds in high yield within a short time. It is also suitable for the synthesis of DNA-encoded compounds using multi-well plates.

Description

Technical Field

[0001] This invention belongs to the field of encoded compound library technology, specifically relating to a method for constructing on-DNA debenzylation in a DNA encoded compound library. Background Technology

[0002] In drug development, especially in the development of new drugs, high-throughput screening targeting biological targets is one of the main methods for rapidly obtaining lead compounds. However, traditional high-throughput screening based on single molecules is time-consuming, requires huge equipment investment, and has a limited number of compounds (millions). Furthermore, the construction of compound libraries requires decades of accumulation, which limits the efficiency and possibility of lead compound discovery. In recent years, DNA-encoded compound library technology (WO2005058479, WO2018166532, CN103882532) has emerged. This technology combines combinatorial chemistry and molecular biology techniques, adding a DNA tag to each compound at the molecular level. It can synthesize compound libraries of up to hundreds of millions in a very short time, becoming the trend of next-generation compound library screening technology. It has begun to be widely used in the pharmaceutical industry, producing many positive effects (Accounts of Chemical Research, 2014, 47, 1247-1255).

[0003] DNA-encoded compound libraries enable the rapid generation of giant compound libraries through combinatorial chemistry, and allow for high-throughput screening of lead compounds, making lead compound screening faster and more efficient than ever before. One of the challenges in constructing DNA-encoded compound libraries is the need for high-yield synthesis of chemically diverse small molecules on DNA. Since DNA requires specific conditions (solvent, pH, temperature, ion concentration) to maintain stability, and the On-DNA reactions used in the construction of DNA-encoded compound libraries also need to achieve high yields, the types of reagents, reaction types, and reaction conditions of chemical reactions performed on DNA (referred to as On-DNA reactions) directly affect the richness and selectivity of DNA-encoded compound libraries. Therefore, developing DNA-compatible chemical reactions has become a long-term exploration and research direction for DNA-encoded compound library technology, directly impacting the application and commercial value of DNA-encoded compound libraries.

[0004] Developing methods for removing benzyl groups from DNA-encoded compounds can enrich the applications of amino and hydroxyl groups, thereby expanding the diversity of compound libraries and increasing the probability of screening effective compounds. However, no methods for removing benzyl groups from DNA-encoded compounds have been reported to date. Therefore, it is hoped that a new synthetic method for removing benzyl groups from DNA-encoded compounds suitable for high-volume multi-well plate operations can be developed to increase the diversity of DNA-encoded compound libraries and further enhance the application value of DNA-encoded compound library technology. Summary of the Invention

[0005] To address the aforementioned issues, this invention develops a method for synthesizing DNA-encoded compound libraries that features stable raw material storage, mild reaction conditions, good substrate versatility, minimal DNA damage, and suitability for batch processing using multi-well plates. This method can rapidly convert On-DNA libraries containing benzyl-protected amino and / or hydroxyl compounds into On-DNA libraries containing amino and / or hydroxyl compounds in a single reaction.

[0006] This invention provides a method for removing benzyl groups from DNA-encoded compounds. The method uses on-DNA containing benzyl-protected amino and / or hydroxyl compounds as raw materials, and reacts them in the presence of a palladium catalyst and a hydrogen source to obtain on-DNA containing amino and / or hydroxyl compounds.

[0007] Among them, the structural formula of the on-DNA containing a benzyl-protected amino or hydroxyl compound is as follows: The structural formula of on-DNA compounds containing amino or hydroxyl groups is:

[0008] In the structural formula, the DNA contains a single-stranded or double-stranded nucleotide chain obtained by polymerizing artificially modified and / or unmodified nucleotide monomers, which is linked to R1 or R3 by one or more chemical bonds or groups.

[0009] The DNA is 10 to 200 base pairs in length.

[0010] In this structural formula, the DNA is connected to R1 or R3 by one or more chemical bonds. A single chemical bond means the DNA is directly connected to R1 or R3; multiple chemical bonds mean the DNA is connected to R1 or R3 by several intervening chemical bonds. For example, the DNA is connected to the amino group of R1 or R3 by a methylene group (-CH2-), i.e., by two chemical bonds; or the DNA is connected to the amino group of R1 or R3 by a carbonyl group (-CO-), also by two chemical bonds; or the DNA is connected to the amino group of R1 or R3 by a methylene carbonyl group (-CH2CO-), also by three consecutive chemical bonds.

[0011] Preferably, the amino group of DNA is linked to R1 or R3 via a carbonyl group (-CO-), a methylene carbonyl group (-CH2CO-), or a -CH(CH3)CO-.

[0012] n is selected from 1, 2, or 3;

[0013] R1 is selected from groups with a molecular weight of less than 1000 that are directly linked to DNA and nitrogen atoms, or it does not exist;

[0014] R2 is selected from hydrogen or a group with a molecular weight of less than 1000 that is directly bonded to a nitrogen atom;

[0015] R3 is selected from groups with a molecular weight of less than 1000 that are directly attached to DNA and hydroxyl oxygen atoms;

[0016] Alternatively, R1 and R2 may be connected to the nitrogen atom directly attached to them to form a heterocycle or aromatic heterocycle.

[0017] R1, R2, and R3 are each independently selected from alkyl, substituted alkyl, 5-10 aryl, substituted 5-10 aryl, 5-10 aryl heterocyclic, substituted 5-10 aryl heterocyclic, C3-C8 cycloalkyl, substituted C3-C8 cycloalkyl, C3-C8 heterocyclic alkyl, and substituted C3-C8 heterocyclic alkyl; wherein the alkyl group is C1-C6. 10 Alkyl; the number of substituents in a substituted alkyl group is one or more; the substituents in a substituted alkyl group are independently selected from halogens, carboxyl groups, nitro groups, and C1-C2 groups. 10 One or more of alkoxy, halophenyl, and phenyl;

[0018] The number of substituents replacing the 5- to 10-membered aryl groups is one or more, and the substituents replacing the 5- to 10-membered aryl groups are independently selected from halogens, oxo groups, cyano groups, nitro groups, carboxyl groups, and C1-C1 groups. 10 Alkoxy, C1-C 10 One or more of alkyl and trifluoromethyl groups;

[0019] The number of substituents replacing the 5-10 membered aromatic heterocyclic group is one or more, and the substituents replacing the 5-10 membered aromatic heterocyclic group are independently selected from halogen, oxo, cyano, nitro, carboxyl, C1-C2 groups. 10 Alkoxy, C1-C 10 One or more of alkyl and trifluoromethyl groups;

[0020] The number of substituents in the C3-C8 cycloalkyl group is one or more, and the substituents in the C3-C8 cycloalkyl group are independently selected from halogen, oxo, cyano, nitro, carboxyl, alkoxy, C1-C8 cycloalkyl groups. 10 One or more of alkyl and trifluoromethyl groups;

[0021] The number of substituents in the substituted C3-C8 heterocyclic alkyl group is one or more, and the substituents in the substituted C3-C8 heterocyclic alkyl group are independently selected from halogen, oxo, cyano, nitro, carboxyl, alkoxy, C1-C8 heterocyclic alkyl groups. 10 One or more of alkyl and trifluoromethyl groups;

[0022] Alternatively, R1 and R2 can form C3-C8 heterocycles or 5-10 membered aromatic heterocycles with the nitrogen atom directly attached to them; the heterocycles or aromatic heterocycles can be further formed by one, two, or three independent R atoms. 1a replace;

[0023] Each R 1a Each group is independently selected from hydrogen, halogen, cyano, oxo, nitro, and -C. 1～10 Alkyl, halogen-substituted -C 1～10 Alkyl, -OC 1～10 alkyl.

[0024] Preferably, R1 is selected from -C 1～6 alkyl.

[0025] The R2 mentioned is selected from -C 1～6 Alkyl, more specifically, R2 is selected from methyl and ethyl.

[0026] Alternatively, R1 and R2 can form a C4-C6 heterocycle or a 5-6 membered aromatic heterocycle with the nitrogen atom directly attached to them; the heterocycle or aromatic heterocycle can be further formed by one, two, or three independent R atoms. 1a replace;

[0027] Each R 1a Each element is independently selected from hydrogen, halogen, oxo, and -C. 1～6 alkyl.

[0028] More specifically, R1 and R2 form with the nitrogen atoms directly bonded to them, including but not limited to...

[0029] Preferably, R3 is selected from 6-membered aryl, substituted 6-membered aryl, 5-10-membered aromatic heterocyclic, and C3-C6 heterocyclic alkyl.

[0030] The number of substituents replacing the 6-membered aryl group is one or more, and the substituents replacing the 6-membered aryl group are independently selected from halogens and C1 to C3 alkoxy groups;

[0031] More specifically: the R3 mentioned includes, but is not limited to,

[0032] Preferably, the On-DNA contains a benzyl-protected amino or hydroxyl compound with the following structural formula:

[0033]

[0034]

[0035] This invention provides a method for removing benzyl groups from DNA-encoded compounds. The method includes the following steps: adding 0.1 to 1000 molar equivalents of palladium catalyst to a solution of an on-DNA containing benzyl-protected amino and / or hydroxyl compounds with a molar equivalent of 1 and a molar concentration of 0.5-5 mM; finally adding 0.1 to 1000 molar equivalents of hydrogen source; and reacting at 10°C to 100°C for 0.5 to 24 hours until the reaction is complete.

[0036] Furthermore, the palladium catalyst is selected from palladium acetate, palladium chloride, palladium hydroxide, tetra(triphenylphosphine)palladium, tris(dibenzylacetone)palladium, bis(dibenzylacetone)palladium, bis(acetonitrile)palladium(II) dichloride, bis(triphenylphosphine)palladium chloride, 1,1'-bis(diphenylphosphino)ferrocene palladium dichloride, bis(benzonitrile)palladium dichloride, 1,4-bis(diphenylphosphine)butane-palladium chloride, [di-tert-butyl(chloride)phosphine]palladium(II) dichloride dimer, bis(methyldiphenylphosphine)palladium(II) dichloride, and benzylbis(triphenylphosphine). Palladium(II) chloride, dihydrodichlorobis(di-tert-butylphosphine-KP)palladium acid (2-), sodium chloride (sodium-2-dicyclohexylphosphine-2',6'-dimethoxy-1,1'-biphenyl-3'-sulfonate)[2-(2'-amino-1,1'-biphenyl)]palladium(II), methanesulfonic acid (2-dicyclohexylphospho-2',6'-dimethoxy-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II), methanesulfonic acid (2-dicyclohexylphospho-2',6'-diisopropoxy) -1,1'-Biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II), Chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II), Chloro(2-dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2-aminoethylphenyl)]palladium(II), Methanesulfonic acid (9,9-dimethyl-4,5-bisdiphenylphosphine) (2'-amino-1,1'-biphenyl-2-yl)palladium(II), (2-dicyclohexylphosphino-N,N-dimethylamino-1,1'-biphenyl)palladium(II), ((4-(N,N-dimethylamino)phenyl)di-tert-butylphosphine (2-amino-1,1'-biphenyl-2-yl)palladium(II), (1,1'-bis(diphenylphosphine)ferrocene (2-amino-1,1'-biphenyl-2-yl)palladium(II);

[0037] Preferably, the palladium catalyst is palladium dichloride.

[0038] Furthermore, the hydrogen source is selected from one of hydrogen, sodium borohydride, sodium cyanoborohydride, trialkylsilane, dialkylsilane, triarylsilane, diarylsilane, and triethylsilane.

[0039] Preferably, the hydrogen source is triethylsilane.

[0040] Furthermore, the reaction is carried out in a solvent, which is any one or a mixture of several of the following aqueous solvents: water, methanol, ethanol, acetonitrile, dimethyl sulfoxide, dimethylformamide, dimethylacetamide, inorganic salt buffer, organic acid buffer, and organic base buffer.

[0041] Preferably, the reaction solvent is a mixed solution of water and dimethylacetamide.

[0042] Further, the reaction temperature is 10℃~100℃; preferably, the reaction temperature is 20℃, 30℃, 40℃, 50℃, 60℃, 70℃, 80℃, 90℃, or 100℃.

[0043] Furthermore, the reaction time is 0.5 to 24 hours; preferably, the reaction time is 1 hour, 2 hours, 4 hours, 8 hours, 10 hours, 16 hours, 18 hours, or 20 hours.

[0044] Further, in the method, the On-DNA contains an equivalent of 1 benzyl-protected amino and / or hydroxyl compound, the palladium catalyst has a molar equivalent of 0.1 to 1000, and the hydrogen source has a molar equivalent of 0.1 to 1000; preferably, the molar equivalent of the palladium catalyst is 0.1, 1, 5, 10, 50, 100, 200, 300, 400, 500, 600, 800, or 1000, and the molar equivalent of the hydrogen source is 0.1, 1, 5, 10, 50, 100, 200, 300, 400, 500, 700, 800, or 1000.

[0045] Most preferably, the molar equivalent of the palladium catalyst is 10 and the molar equivalent of the hydrogen source is 700.

[0046] Furthermore, the reaction is carried out in the following order: first, add the On-DNA containing a benzyl-protected amino and / or hydroxy compound, then add the palladium catalyst, and finally add the hydrogen source.

[0047] Furthermore, the method is used for batch operations of perforated plates.

[0048] Furthermore, the method is used for the synthesis of DNA-encoded compound libraries for multi-well plates.

[0049] This invention provides a method for removing benzyl groups from DNA-encoded compound libraries. It can be widely applied to various on-DNA compounds containing benzyl-protected amino and / or hydroxyl groups. The method offers high yields, produces a single product, can be carried out in a mixed aqueous phase using organic solvents and water, is simple to operate, environmentally friendly, and suitable for the synthesis of DNA-encoded compound libraries using multi-well plates.

[0050] Regarding the definition of terms used in this invention: Unless otherwise stated, the initial definitions provided for groups or terms herein apply to the groups or terms used throughout this specification; for terms not specifically defined herein, the meanings that a person skilled in the art would give them should be given based on the disclosure and context.

[0051] "Substitution" refers to the replacement of hydrogen atoms in a molecule by other different atoms or molecules.

[0052] The minimum and maximum carbon atom content in hydrocarbon groups are indicated by prefixes, for example, prefixes (Ca~C). b Alkyl groups indicate any alkyl group containing one to two carbon atoms ("a" to "b"). Therefore, for example, C1 to C2... 12 Alkyl groups are straight-chain or branched alkyl groups containing 1 to 12 carbon atoms.

[0053] Alkyl refers to a straight-chain or branched hydrocarbon group in an alkane molecule, such as methyl-CH3, ethyl-CH2CH3, or methylene-CH2-. The alkyl group can also be part of other groups, such as C1-C6 alkoxy groups or C1-C6 alkylamino groups.

[0054] The halogen is fluorine, chlorine, bromine or iodine.

[0055] Alkoxy refers to an alkyl group that is attached to an oxygen atom to form a substituent, such as methoxy, which is -OCH3.

[0056] A halophenyl group is a group formed when the hydrogen atom on a phenyl group is replaced by a halogen.

[0057] Cycloalkyl refers to a saturated or partially saturated cyclic group having multiple carbon atoms and no heterocyclic atoms, and having a single ring or multiple rings (including fused, bridged and spirocyclic systems).

[0058] Heterocyclic groups are saturated or unsaturated monocyclic or polycyclic hydrocarbon groups carrying at least 3 to 8 atoms selected from O, S, and N.

[0059] 5- to 10-membered aryl groups refer to aromatic single or multiple cyclic groups composed of carbon atoms and lacking heteroatoms.

[0060] 5- to 10-membered aromatic heterocyclic groups refer to single or multiple cyclic groups composed of 5 to 10 C, O, S, N atoms, etc., which have aromatic properties.

[0061] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions or alterations can be made without departing from the basic technical concept of the present invention.

[0062] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. Attached Figure Description

[0063] Figure 1 The conversion rate distribution of On-DNA compounds containing amino or hydroxyl groups obtained in Example 2 of this invention is shown in the figure. Detailed Implementation Plan

[0064] The raw materials and equipment used in this invention are all known products, obtained by purchasing commercially available products.

[0065] In this invention, DNA-NH2 refers to a DNA structure with a -NH2 linker formed by single-stranded or double-stranded DNA and a linker group, such as the DNA-NH2 structure of "compound 1" in WO2005058479. Other examples include the following DNA structures:

[0066]

[0067] Where A is adenine, T is thymine, C is cytosine, and G is guanine.

[0068] Example 1: Benzyl removal of benzyl-protected amino or hydroxyl compounds from on-DNA

[0069] Step 1: Synthesis of On-DNA compounds containing benzyl-protected amino or hydroxyl groups

[0070]

[0071] DNA-NH2(HP) was dissolved in 250 mM, pH 9.4 borate buffer to prepare a 1 mM DNA solution (1 equivalent). N-benzyl o-aminobenzoic acid (50 equivalent, 0.2 M dissolved in dimethylacetamide), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (50 equivalent, 0.4 M dissolved in dimethylacetamide), and N,N-diisopropylamine (50 equivalent, 0.4 M dissolved in dimethylacetamide) were mixed thoroughly and then added to the DNA solution. The mixture was stirred thoroughly and reacted at 25°C for 1 hour.

[0072] After the reaction, ethanol precipitation was performed: 10% of the total volume of 5M sodium chloride solution was added to the solution after the reaction, followed by 3 times the total volume of anhydrous ethanol. After shaking and homogenization, the reaction was placed in dry ice and frozen for 0.5 hours. Then, it was centrifuged at 12000 rpm at low temperature (4℃) for half an hour. The supernatant was discarded, and the residual precipitate was freeze-dried and then dissolved in deionized water to obtain solutions of compounds 1 and 2. After quantification by OD, the conversion rates of compounds 1 and 2 were confirmed by LC-MS to be 95% and 96%, respectively.

[0073] Step 2: On-DNA debenzylation

[0074]

[0075] Compounds 1 and 2 were prepared into a 1 mM DNA solution (1 equivalent) using a 500 mM sodium acetate / acetic acid buffer solution at pH 5.5. Palladium dichloride (10 equivalents, 50 mM dissolved in 5 M sodium chloride aqueous solution), triethylsilane (700 equivalents, 2 M dissolved in dimethylacetamide), and acetic acid (200 equivalents, 1 M dissolved in water) were added sequentially to the solution. The mixture was thoroughly mixed and reacted at 60 °C for 30 minutes.

[0076] After the reaction was complete, sodium diethyldithiocarbamate (100 equivalents, 0.5 M dissolved in water) was added to the reaction system, mixed thoroughly, and reacted at 60°C for 15 minutes. After the reaction was complete, 10% by volume of 5 M sodium chloride solution was added to the resulting solution, followed by 3 times the total volume of anhydrous ethanol. After shaking thoroughly, the reaction mixture was placed on dry ice and frozen for 0.5 hours. Then, it was centrifuged at 12000 rpm at low temperature (4°C) for half an hour. The supernatant was discarded, and the remaining precipitate was freeze-dried and then dissolved in deionized water to obtain solutions of compounds 1-1 and 2-1. After quantification by OD, the conversion rates of compounds 1-1 and 2-1 were confirmed by LC-MS to be 98% and 96%, respectively.

[0077] Example 2: Method for removing benzyl groups from amino or hydroxyl compounds protected by benzyl groups in on-DNA

[0078] Sixteen on-DNA compounds containing benzyl-protected amino or hydroxyl groups were prepared into 1 mM DNA solutions (1 equivalent) using 500 mM sodium acetate / acetic acid buffer solution at pH 5.5. Palladium dichloride (10 equivalent, 50 mM dissolved in 5 M sodium chloride aqueous solution), triethylsilyl hydride (700 equivalent, 2 M dissolved in dimethylacetamide), and acetic acid (200 equivalent, 1 M dissolved in water) were added sequentially to the solutions. The mixtures were thoroughly mixed and reacted at 60 °C for 30 minutes.

[0079] After the reaction was complete, sodium diethyldithiocarbamate (100 equivalents, 0.5 M dissolved in water) was added to the reaction system, mixed thoroughly, and reacted at 60°C for 15 minutes. After the reaction was complete, 10% by volume of 5 M sodium chloride solution was added to the resulting solution, followed by 3 times the total volume of anhydrous ethanol. After shaking thoroughly, the reaction mixture was placed on dry ice for 0.5 hours, then centrifuged at 12000 rpm at low temperature (4°C) for half an hour. The supernatant was discarded, and the remaining precipitate was freeze-dried and then dissolved in deionized water to obtain the compound solution. After quantification by OD, the conversion rate of the compound was confirmed by LC-MS. For specific conversion rates, see [link to relevant documentation]. Figure 1 .

[0080] Table 1: Structural Formulas of Raw Materials and Products Used in Example 2

[0081]

[0082]

[0083]

[0084] This method exhibits good universality in both benzyl-protected hydroxyl compounds and benzyl-protected amino compounds.

[0085] In summary, this invention successfully achieved on-DNA benzyl removal by controlling factors such as solvent, temperature, and pH during the reaction, in the presence of a palladium catalyst and a hydrogen source. This method has a wide substrate applicability, can be carried out in a mixed aqueous phase of organic solvent / aqueous phase, is simple to operate, environmentally friendly, and suitable for the synthesis of DNA-encoded compound libraries using multi-well plates.

Claims

1. A method for removing benzyl groups from a DNA-encoded compound, characterized in that: On-DNA containing amino and / or hydroxyl compounds protected by benzyl groups was obtained by reacting the compounds with amino and / or hydroxyl groups in the presence of a palladium catalyst and a hydrogen source. The on-DNA contains a benzyl-protected amino or hydroxy compound with the following structural formula: or On-DNA contains the structural formula of an amino or hydroxyl compound. or ; In the structural formula, the DNA contains a single-stranded or double-stranded nucleotide chain obtained by polymerizing artificially modified and / or unmodified nucleotide monomers, which is linked to R1 or R3 by one or more chemical bonds or groups. n is selected from 1, 2, or 3; R1, R2, and R3 are each independently selected from alkyl, substituted alkyl, 5-10 aryl, substituted 5-10 aryl, 5-10 aryl heterocyclic, substituted 5-10 aryl heterocyclic, C3-C8 cycloalkyl, substituted C3-C8 cycloalkyl, C3-C8 heterocyclic alkyl, and substituted C3-C8 heterocyclic alkyl; wherein the alkyl group is C1-C6. 10 Alkyl; the number of substituents in a substituted alkyl group is one or more; the substituents in a substituted alkyl group are independently selected from halogens, carboxyl groups, nitro groups, and C1-C2 groups. 10 One or more of alkoxy, halophenyl, and phenyl; The number of substituents replacing the 5-10 aryl group is one or more, and the substituents replacing the 5-10 aryl group are independently selected from halogen, oxo, cyano, nitro, carboxyl, C1-C2 groups. 10 Alkoxy, C1~C 10 One or more of alkyl and trifluoromethyl groups; The number of substituents replacing the 5-10 membered aromatic heterocyclic group is one or more, and the substituents replacing the 5-10 membered aromatic heterocyclic group are independently selected from halogen, oxo, cyano, nitro, carboxyl, C1~C 10 Alkoxy, C1~C 10 One or more of alkyl and trifluoromethyl groups; The number of substituents in the C3-C8 cycloalkyl group is one or more, and the substituents in the C3-C8 cycloalkyl group are independently selected from halogen, oxo, cyano, nitro, carboxyl, alkoxy, C1-C8 cycloalkyl groups. 10 One or more of alkyl and trifluoromethyl groups; The number of substituents in the substituted C3-C8 heterocyclic alkyl group is one or more, and the substituents in the substituted C3-C8 heterocyclic alkyl group are independently selected from halogen, oxo, cyano, nitro, carboxyl, alkoxy, C1-C 10 One or more of alkyl and trifluoromethyl groups; Alternatively, R1 and R2 can form a C3-C8 heterocycle or a 5-10 membered aromatic heterocycle with the nitrogen atom directly attached to them; the heterocycle or aromatic heterocycle is bounded by one, two, or three independent R atoms. 1a replace; Each R 1a Each group is independently selected from hydrogen, halogen, cyano, oxo, nitro, and -C. 1~10 Alkyl, halogen-substituted -C 1~10 Alkyl, -OC 1~10 alkyl; The method includes the following steps: adding 0.1 to 1000 molar equivalents of palladium catalyst to a solution of On-DNA containing benzyl-protected amino and / or hydroxyl compounds with a molar equivalent of 1 and a molar concentration of 0.5-5 mM, and finally adding 0.1 to 1000 molar equivalents of hydrogen source, and reacting at 10℃ to 100℃ for 0.5 to 24 hours.

2. The method according to claim 1, characterized in that: The palladium catalyst is selected from palladium acetate, palladium dichloride, palladium hydroxide, tetra(triphenylphosphine)palladium, tris(dibenzylacetone)palladium, bis(dibenzylacetone)palladium, bis(acetonitrile)palladium dichloride (II), bis(triphenylphosphine)palladium dichloride, 1,1'-bis(diphenylphosphino)ferrocene palladium dichloride, bis(benzonitrile)palladium dichloride, 1,4-bis(diphenylphosphine)butane-palladium chloride, [di-tert-butylphosphine]palladium dichloride (II) dimer, bis(methyldiphenylphosphine)palladium dichloride (II), benzylbis(triphenylphosphine)palladium dichloride (II), dihydrodichlorodi(di-tert-butylphosphine-KP)palladium acid (2-), chloro(sodium-2-dicyclohexylphosphine-2',6'-dimethoxy-1,1'-biphenyl-3'-sulfonate)[2-(2'-amino-1,1'-... Palladium(II), Methanesulfonic acid (2-dicyclohexylphosphino-2',6'-dimethoxy-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II), Methanesulfonic acid (2-dicyclohexylphosphino-2',6'-diisopropyl-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II), Chlorine (2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II), Chlorine (2-dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II), Chlorine (2-dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl) The following are one or more of the following: benzene[2-(2-aminoethylphenyl)]palladium(II), methanesulfonic acid (9,9-dimethyl-4,5-bisdiphenylphosphineoxanthracene)(2'-amino-1,1'-biphenyl-2-yl)palladium(II), methanesulfonic acid (2-dicyclohexylphosphino-N,N-dimethylamino-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II), methanesulfonic acid [4-(N,N-dimethylamino)phenyl]di-tert-butylphosphine(2-amino-1,1'-biphenyl-2-yl)palladium(II), and methanesulfonic acid-1,1'-bis(diphenylphosphine)ferrocene(2-amino-1,1'-biphenyl-2-yl)palladium(II).

3. The method according to claim 1, characterized in that: The hydrogen source is selected from one of the following: hydrogen, sodium borohydride, sodium cyanoborohydride, trialkylsilane, dialkylsilane, triarylsilane, diarylsilane, and triethylsilane.

4. The method according to claim 1, characterized in that: The reaction is carried out in a solvent, which is any one or a mixture of several of the following aqueous solvents: water, methanol, ethanol, acetonitrile, dimethyl sulfoxide, dimethylformamide, dimethylacetamide, inorganic salt buffer, organic acid buffer, and organic base buffer.

5. The method according to claim 1, characterized in that: The reaction temperatures are 20℃, 30℃, 40℃, 50℃, 60℃, 70℃, 80℃, 90℃, and 100℃.

6. The method according to claim 1, characterized in that: The reaction times are 1 hour, 2 hours, 4 hours, 8 hours, 10 hours, 16 hours, 18 hours, and 20 hours.

7. The method according to claim 1, characterized in that: In the method, the on-DNA contains a benzyl-protected amino and / or hydroxyl compound with a molar equivalent of 1, the palladium catalyst has a molar equivalent of 0.1, 1, 5, 10, 50, 100, 200, 300, 400, 500, 600, 800, or 1000, and the hydrogen source has a molar equivalent of 0.1, 1, 5, 10, 50, 100, 200, 300, 400, 500, 700, 800, or 1000.

8. The method according to any one of claims 1-7, characterized in that, The method is used for batch operations of multi-hole plates.

9. The method according to any one of claims 1-7, characterized in that, The method is used for the synthesis of DNA-encoded compound libraries for multi-well plates.

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