Synthesis method and application of on-DNA carbonyl compound

Abstract

The invention belongs to the technical field of DNA coding molecules, and provides a synthesis method of an on-DNA carbonyl compound, which comprises: providing an on-DNA amino compound and an on-DNA hydroxyl compound, the on-DNA amino compound comprising a DNA fragment and an amino group coupled to the DNA fragment, and the on-DNA hydroxyl compound comprising a DNA fragment and a hydroxyl group coupled to the DNA fragment; and reacting the on-DNA amino compound and / or the on-DNA hydroxyl compound with laccase and an auxiliary catalyst to obtain the on-DNA carbonyl compound, namely the on-DNA carbonyl compound containing a DNA fragment and carbonyl coupled with the DNA fragment. The synthesis method has the advantages of mild conditions, environmental friendliness and small damage to DNA, and is suitable for construction of a DNA coding molecular library, fluorescence labeled molecule coupling performed by using an on-DNA carbonyl reaction handle, DNA modification coupling and functional modification in targeted drug delivery.

Description

Technical Field

[0001] The invention belongs to the technical field of DNA encoding molecule library, and specifically relates to a synthesis method of an on-DNA carbonyl compound and application thereof. Background Art

[0002] DNA molecules have important research value and broad application prospects in the fields of biomedicine, molecular biology and nucleic acid chemistry. By chemically modifying DNA molecules, their functions and application range can be significantly expanded, such as for the construction of DNA-encoded molecular libraries, targeted drug delivery, molecular probe design and gene expression regulation. However, the methods for efficient and selective chemical modification of DNA molecules are still limited.

[0003] DNA-aldehyde conjugates are a class of on-DNA carbonyl compounds that modify aldehyde groups at the ends and / or any positions in the middle of DNA. They are widely used in the construction of DNA-encoded molecular libraries (DELs), biomarkers, and drug screening. As a reactive center, DNA-aldehydes can react with amines, hydrazines, or other nucleophiles to generate covalently bound DNA-modified compounds. With the development of DEL technology and biomarker technology, the preparation of DNA-aldehydes has become a hot topic of research. At present, the main methods for preparing DNA-aldehydes include the following: ① By introducing an alcohol group (-CH 2 OH) structure, and then use an oxidant (such as Pinnick oxidation system, potassium permanganate, TEMPO, etc.) to oxidize the alcohol to an aldehyde. ② By introducing aldehyde-functionalized phosphoramide compounds during DNA synthesis, the aldehyde group is directly modified at the end of the DNA chain. ③ By introducing a carboxyl group (-COOH) at the end of the DNA, and then using sodium diborohydride (NaBH 4 ) or other reducing agents to reduce the carboxyl group to an aldehyde group. ④ Specifically convert the amino or hydroxyl groups on certain nucleosides (such as cytosine or uracil) into aldehyde groups through chemical reactions. ⑤ By introducing a clickable precursor (such as an azide or alkyne group) at the end of the DNA chain, and then converting it into an aldehyde group through subsequent reactions. Although the existing DNA-aldehyde preparation methods have their own characteristics, there are still the following technical defects: the destructiveness of the reaction conditions to the DNA molecular structure, the large amount of by-products generated, the limited yield, and the complex synthesis steps and high costs. Therefore, there is an urgent need for a new, simple, efficient and DNA structure-friendly method for preparing high-purity DNA-aldehyde compounds to meet the development needs of DEL technology and biomarker technology.

[0004] Amino and hydroxyl groups are common modification groups in DNA molecules. Oxidation of these groups to aldehyde groups can provide active sites for further chemical reactions, such as Schiff base formation or subsequent coupling reactions. Existing oxidation methods mostly rely on chemical reagents (such as peroxides or heavy metal catalysts), which may cause irreversible damage to DNA molecules and have poor selectivity.

[0005] Therefore, developing an efficient, selective and mild method for synthesizing on-DNA carbonyl compounds to achieve functional modification on DNA molecules has important scientific significance and application value for the development of new drugs. Summary of the invention

[0006] In view of this, the purpose of the present invention is to provide a method for synthesizing on-DNA carbonyl compounds to solve the problems of harsh oxidation reaction conditions, poor selectivity and possible damage to the DNA backbone in the prior art. In addition, the present invention also aims to expand the application scope of DNA chemical modification, such as the construction of DNA encoding molecule libraries, coupling of fluorescent labeled molecules using on-DNA carbonyl reaction handles, DNA modification coupling and functional modification in targeted drug delivery, so as to promote the development of related drugs.

[0007] In order to achieve the above-mentioned invention object, the technical solution adopted by the present invention is as follows:

[0008] In a first aspect, the present invention provides a method for synthesizing an on-DNA carbonyl compound, comprising the following steps:

[0009] Providing an on-DNA amine compound and an on-DNA hydroxyl compound, wherein the on-DNA amine compound comprises a DNA fragment and an amine group coupled to the DNA fragment, and the on-DNA hydroxyl compound comprises a DNA fragment and a hydroxyl group coupled to the DNA fragment;

[0010] The on-DNA amine compound and / or the on-DNA hydroxyl compound is reacted with laccase and an auxiliary catalyst to obtain an on-DNA carbonyl compound.

[0011] Laccase, as a green biocatalyst, has the advantages of mildness, high selectivity and environmental friendliness, and has received extensive attention in the fields of organic synthesis and biomolecule modification in recent years. The above method provided by the present invention uses laccase to oxidize the amino / amine group coupled to the DNA fragment in the on-DNA compound molecule, with mild conditions, high efficiency and high selectivity, overcoming the problem of irreversible damage to the DNA main chain in the oxidation process of traditional chemical reagents, and having the advantages of simple operation, environmental friendliness and high efficiency.

[0012] In some embodiments, the pH of the reaction is 4.0-7.5; and / or

[0013] The reaction temperature is 25°C-37°C; and / or

[0014] The reaction time is 2-48 hours.

[0015] In some embodiments, the amount of laccase used is 1 to 100 U / mL.

[0016] In some embodiments, the auxiliary catalyst is selected from TEMPO (Chinese name: 2,2,6,6-tetramethylpiperidine oxide, English name: 2,2,6,6-Tetramethylpiperidine-1-oxyl), HBT (Chinese name: 1-hydroxybenzotriazole, English name: 1-Hydroxybenzotriazole), ABTS (Chinese name: 2,2'-hydrazine bis (3-ethylbenzothiazoline-6-sulfonic acid), English name: 2,2'-Azino-bis (3-ethylbenzothiazoline-6-sulfonicacid), NaClO (Chinese name: sodium hypochlorite), NaClO 2 (sodium chlorite); and / or

[0017] The concentration of the auxiliary catalyst is 0.01 mmol / L to 1 mol / L.

[0018] In some embodiments, the on-DNA amine compound is selected from at least one of Compounds 1 to 59:

[0019]

[0020]

[0021] and / or

[0022] The on-DNA hydroxy compound is selected from at least one of Compound 60 to Compound 83:

[0023]

[0024] The above method provided by the present invention can realize the selective oxidation of amino / amine / hydroxyl groups on DNA under biocompatible conditions, retain the integrity of DNA, and provide active carbonyl groups (such as aldehyde groups) as reaction sites for subsequent chemical modifications. Specifically, the present invention realizes the selective oxidation of amino / amine / hydroxyl groups on DNA by using laccase and an auxiliary catalyst (such as TEMPO), overcomes the problem of DNA damage caused by traditional chemical reagents, and has the advantages of simple operation, environmental friendliness and high efficiency.

[0025] In some embodiments, the synthesis method comprises the following steps:

[0026] S11, providing an on-DNA amine compound as shown in the general formula (I):

[0027]

[0028] S12, reacting the on-DNA amine compound in a reaction system containing laccase and an auxiliary catalyst to obtain an on-DNA carbonyl compound as shown in the general formula (II):

[0029]

[0030] in, is the connection key,

[0031] R 1 is selected from one of hydrogen, alkyl, cycloalkyl, alkenyl and aryl, wherein the substituents on the aryl are independently selected from H, C 1 -C 6 At least one of alkyl, halogen, nitro, amino, amine, alkoxy, and cyano,

[0032] R 2 and R 3 Each independently selected from hydrogen, alkyl, halogenated alkyl, aryl, and other groups, wherein the other groups contain at least one of alkyl, acyl, and ester groups, and the substituents on the aryl groups are each independently selected from H, C 1 -C 6 At least one of alkyl, halogen, nitro, amino, amine, alkoxy, and cyano,

[0033] R 4 A group selected from at least one of an alkyl group, a halogenated alkyl group, a cycloalkyl group, an aryl group, and a heteroaryl group, wherein the substituents on the aryl group and the heteroaryl group are independently selected from H, C 1 -C 6 At least one of alkyl, halogen, nitro, amino, amine, alkoxy and cyano.

[0034] The above method provided by the present invention is suitable for oxidizing primary amines, secondary amines and tertiary amines coupled to DNA fragments in on-DNA compound molecules. For primary amines, aldehyde groups are generated after oxidation; for secondary amines and tertiary amines, oxidation can be achieved by a suitable catalyst combination to generate corresponding oxidation products, and the specific structure depends on the substituents on the amines. Specifically, taking the auxiliary catalyst as TEMPO and the amine group coupled to the DNA fragment in the on-DNA compound molecule as a primary amine as an example, the reaction mechanism of the above method provided by the present invention for oxidizing on-DNA amino compounds includes: laccase catalyzes the oxidation of TEMPO in the presence of oxygen to generate an oxidized free radical TEMPO + ; Then, the amino group is catalytically oxidized to form an imine intermediate; thereafter, the imine intermediate is hydrolyzed to form an aldehyde group without destroying the stability of the DNA backbone.

[0035] In a further embodiment, the on-DNA amine compound is selected from Compound 1 to Compound 59.

[0036] In a further embodiment, the reaction has a pH value of 4.0-7.5, a temperature of 25°C-37°C, and a reaction time of 2-48 hours; and / or

[0037] The reaction system contains sodium acetate buffer, and the pH value of the sodium acetate buffer is 4.0-7.5.

[0038] In a further embodiment, the laccase is used in an amount of 1 to 100 U / mL; and / or

[0039] The auxiliary catalyst is selected from TEMPO, HBT, ABTS, NaClO, NaClO 2 At least one of; and / or

[0040] The concentration of the auxiliary catalyst is 0.1 mmol / L to 10 mmol / L.

[0041] In other embodiments, the synthesis method comprises the following steps:

[0042] S21, providing an on-DNA hydroxy compound as shown in the general formula (III):

[0043]

[0044] S22, reacting the on-DNA hydroxyl compound in a reaction system containing laccase and an auxiliary catalyst to obtain an on-DNA carbonyl compound as shown in the general formula (IV):

[0045]

[0046] in, is the connection key;

[0047] R 5 is selected from one of aryl and heteroaryl, and the substituents on the aryl and heteroaryl are independently selected from H, C 1 -C 6 At least one of alkyl, halogen, nitro, amino, amine, alkoxy and cyano.

[0048] The above method provided by the present invention is also applicable to the oxidation of aromatic methylene alcohols (such as benzyl alcohol) coupled with DNA fragments in the on-DNA compound molecules, but cannot be used to oxidize alkyl alcohols (such as methanol) coupled with DNA fragments in the on-DNA compound molecules. Specifically, taking the on-DNA compound molecules formed by the coupling of auxiliary catalyst TEMPO, benzyl alcohol and DNA fragments as an example, the reaction mechanism of the above method provided by the present invention for oxidizing on-DNA hydroxyl compounds includes: laccase catalyzes the oxidation of TEMPO in the presence of oxygen to generate oxidized free radical TEMPO + ; Then, TEMPO + The α-hydrogen atom (the hydrogen attached to the hydroxyl group) is extracted from the alcohol molecule to generate an alcohol free radical intermediate, which is further oxidized to release a proton.

[0049] In a further embodiment, the on-DNA amine compound is selected from Compound 1 to Compound 59;

[0050] The on-DNA hydroxy compound is selected from Compound 60 to Compound 83.

[0051] In a further embodiment, the reaction has a pH value of 4.0-7.5, a temperature of 25°C-37°C, and a reaction time of 2-48 hours; and / or

[0052] The reaction system contains sodium acetate buffer, and the pH value of the sodium acetate buffer is 4.0-7.5.

[0053] In a further embodiment, the laccase is used in an amount of 1 to 100 U / mL; and / or

[0054] The auxiliary catalyst is selected from TEMPO, HBT, ABTS, NaClO, NaClO 2 At least one of; and / or

[0055] The concentration of the auxiliary catalyst is 0.1 mmol / L to 10 mmol / L.

[0056] Compared with the prior art, the method provided by the present invention has the following advantages:

[0057] (1) High selectivity: only oxidizes amino / amine / hydroxyl groups without affecting other sensitive groups in the DNA molecule, thus ensuring the integrity of the DNA molecule;

[0058] (2) Mild conditions: Laccase is a green biocatalyst with mild reaction conditions, high selectivity, and environmental friendliness. The reaction is carried out under biocompatible conditions to avoid degradation of DNA molecules;

[0059] (3) Wide applicability: It is suitable for a variety of DNA fragments and amino / amine / hydroxyl modifications and has good application prospects.

[0060] In the second aspect, the on-DNA carbonyl compounds obtained by the above-mentioned synthesis method are used in constructing DNA-encoded molecule libraries, coupling fluorescent labeling molecules, and functionally modifying targeted drug delivery.

[0061] The method provided in the present invention can achieve selective oxidation of amino groups on DNA under biocompatible conditions, retain the integrity of DNA and provide active aldehyde groups as reaction sites for subsequent chemical modifications. On this basis, the application scope of DNA chemical modification can be further expanded, such as the construction of DNA-encoded molecular libraries, coupling of fluorescent labeled molecules and functional modification in targeted drug delivery, which has important scientific significance and application value for new drug research and development. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] Figure 1 It is the reaction mechanism diagram of Example 1 and Example 2.

[0063] Figure 2 This is the LC-MS detection result of compound 1 in Example 1 after the reaction is completed.

[0064] Figure 3 This is the ultraviolet absorption spectrum of the fluorescently labeled compound 1 after the reaction with laccase in Example 1 is completed.

[0065] Figure 4 This is a reaction schematic diagram of an implementation method of the coupling reaction between the on-DNA aldehyde compound and the target molecule (such as a small molecule drug) in Example 4. DETAILED DESCRIPTION

[0066] In the description of the present invention, the compounds and their derivatives involved are named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, located in Columbus, Ohio) naming system, and the specific compound groups involved are described and explained as follows:

[0067] "Alkyl" refers to a type of saturated chain hydrocarbon group containing only carbon and hydrogen atoms, having a straight carbon chain and / or a branched carbon chain, including but not limited to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, isopentyl, hexyl, etc. In the present invention, the number of carbon atoms in the alkyl group is preferably 1-6. In some specific embodiments, the number of carbon atoms in the alkyl group is 1, 2, 3, 4, 5 or 6.

[0068] "Cycloalkyl" refers to a class of saturated hydrocarbon groups containing cyclic structures such as single ring, linked ring, condensed ring, spiro ring and bridged ring in the molecule, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc. In the present invention, the number of carbon atoms in the cycloalkyl group is preferably 3-6. In some specific embodiments, the number of carbon atoms in the cycloalkyl group is 3, 4, 5 or 6.

[0069] "Alkoxy" refers to an alkyl group directly bonded to an oxygen atom, including but not limited to methoxy, ethoxy, propoxy, butoxy, isobutoxy, tert-butoxy, etc. In the present invention, the number of carbon atoms in the alkoxy group is preferably 1-6. In some specific embodiments, the number of carbon atoms in the alkoxy group is 1, 2, 3, 4, 5 or 6.

[0070] "Aryl" refers to any functional group or substituent derived from a simple aromatic ring, which may be monocyclic or polycyclic, including but not limited to phenyl, naphthyl, phenanthrenyl, anthraquinone, etc. The substituents in substituted aryl may be C 1～6 Alkyl, C 1～6 Cycloalkyl, hydroxy, halogen, alkoxy, trifluoromethyl, amino, aldehyde, ester, amide, cyano, etc.

[0071] "Phenyl" refers to a group with a benzene ring as a functional group, such as C 6 H 5 -, the phenyl group may be substituted benzene or unsubstituted benzene. The phenyl group of the present invention may be unsubstituted phenyl group or substituted phenyl group, and the substituent in the substituted phenyl group may be selected from C 1～6 Alkyl, C 1～6 Heteroalkyl, C 1～6 Cycloalkyl, C 1～6 Heterocycloalkyl, hydroxy, halogen, alkoxy, trifluoromethyl, amino, aldehyde, ester, amide, cyano, etc.

[0072] "Heterocyclic group" refers to a group containing heteroatoms such as N, O, S, P, etc. in the ring structure, including aliphatic heterocycles and aromatic heterocycles, including but not limited to thiophene, furan, pyrrole, pyrazole, pyridine, thiazole, pyridopyrrole, oxo-tetrahydroimidazole, etc.

[0073] "Halogen" refers to the elements of Group VIIA in the periodic table, including chlorine (Cl), bromine (Br), iodine (I) and other elements.

[0074] "Amino" refers to NH 2 -, "amine group" refers to NH 2 - is a group formed when at least one hydrogen atom in the

[0075] "Hydroxyl" refers to a group consisting only of O and H, represented by -OH.

[0076] "Cyano" refers to a group consisting only of C and N, represented by -CN.

[0077] "Nitro" refers to a group composed of N and O, represented by -NO 2 .

[0078] The "ester group" refers to a group containing an ester bond.

[0079] "Aldehyde" refers to a group consisting of a carbonyl carbon and a hydrogen bonded together, represented by -CHO.

[0080] "Haloalkyl" refers to an alkyl group in which a hydrogen atom is replaced by a halogen atom, such as -CF 3 .

[0081] In order to make the above implementation details and operations of the present invention clearly understood by those skilled in the art, and to significantly reflect the improved performance of the synthesis method of the on-DNA carbonyl compound and its use in the embodiments of the present invention, the implementation of the present invention is illustrated by examples below.

[0082] The reaction mechanism involved in the following examples is as follows Figure 1 shown.

[0083] Example 1

[0084] In this example, according to the following chemical reaction, the amine group coupled to the DNA molecule in the on-DNA amine compound is oxidized to the corresponding carbonyl group under mild biocompatible conditions to form an aldehyde / ketone.

[0085]

[0086] 1. Reagents and Materials

[0087] Amino-modified DNA fragments (20-50 bases in length): Compounds 1-59 are representative on-DNA amine compounds.

[0088] Laccase: 1-100U / mL.

[0089] Co-catalyst: TEMPO (0.1-10 mM).

[0090] Buffer: 200 mM sodium acetate buffer (pH 5.5).

[0091] 2. Operation steps

[0092] 200 pmol of amino-modified DNA fragments were dissolved in 16 μL buffer, laccase was added to a final concentration of 0.02 U / μL, and TEMPO was added to a final concentration of 10 mM. The reaction mixture was placed at 25°C for 24 hours.

[0093] (1) After the reaction was completed, the product was detected by LC-MS to confirm the conversion rate of amino group to aldehyde group and verify the change of product molecular weight. The results are shown in Table 1.

[0094] Table 1 shows the chemical structures of representative on-DNA amine compounds and their conversion rates and molecular weight data.

[0095] Table 1

[0096]

[0097]

[0098]

[0099]

[0100]

[0101]

[0102] in, Figure 2 The LC-MS test results of compound 1 in Table 1 after the reaction were completed. As shown in the results, the amino group coupled to the DNA fragment in the on-DNA amine compound was effectively oxidized with a conversion rate of more than 90%, and the molecular weight of the oxidation product was consistent with the theoretical value of the aldehyde group.

[0103] (2) Fluorescently label the reaction product of this example to confirm the formation of aldehyde groups

[0104] ① Reagents and materials

[0105] Fluorescent probe: Dinitrophenylhydrazine.

[0106] Catalyst: dimethyl phthalate (DMAP). DMAP accelerates the condensation reaction by activating the aldehyde group.

[0107] ②Operation steps

[0108] After the reaction is completed, the reaction product of this example (e.g., on-DNA aldehyde compound, final concentration 1 μM) is mixed with dinitrophenylhydrazine (final concentration 100 μM), and the catalyst DMAP is added to a final concentration of 10 mM, and the mixture is reacted in a phosphate buffer at pH 7.5 at 37° C. for 4 hours. The fluorescently labeled DNA product is separated and purified using HPLC, and the success of the labeling is verified by the UV absorption spectrum. Figure 3 This is the ultraviolet absorption spectrum of fluorescent labeling after the reaction of compound 1 in Table 1 with laccase is completed. As shown in the results, by scanning the absorption of 300-600nm, it is observed that the absorption around 400nm is significantly enhanced after labeling, indicating that the labeling is successful.

[0109] Example 2

[0110] In this embodiment, the hydroxyl group coupled to the DNA molecule in the on-DNA hydroxyl compound is modified and oxidized into the corresponding aldehyde group or ketone group under mild biocompatible conditions according to the following chemical reaction. 5 is one selected from aryl and heteroaryl, and the substituents on the aryl and heteroaryl are independently selected from H, C 1 -C 6 One of alkyl, halogen and nitro.

[0111]

[0112] 1. Reagents and Materials

[0113] Hydroxyl-modified DNA fragments (20-50 bases in length): Compounds 60-90 are representative on-DNA hydroxyl compounds.

[0114] Laccase: 1-100U / mL.

[0115] Co-catalyst: TEMPO (0.1-10 mM).

[0116] Buffer: 200 mM sodium acetate buffer (pH 5.5).

[0117] 2. Operation steps

[0118] 200 pmol of hydroxyl-modified DNA fragments were dissolved in 16 μL buffer, laccase was added to a final concentration of 0.02 U / μL, and TEMPO was added to a final concentration of 10 mM. The reaction mixture was placed at 25°C for 24 hours. After the reaction, the product was detected by LC-MS to confirm the conversion rate of hydroxyl oxidation to aldehyde group and verify the change in product molecular weight, confirming the formation of aldehyde group.

[0119] Table 2 shows the chemical structures of representative on-DNA hydroxyl compounds and their conversion rates and molecular weight data.

[0120] Table 2

[0121]

[0122]

[0123]

[0124] Example 3

[0125] This example refers to the method of Example 1 and selects Compound 1 in Table 1 to further explore the effect of reaction conditions on reaction efficiency.

[0126] Table 3 shows the results of the study. As shown in the results, the reaction conditions have an effect on the reaction yield, and the optimal conditions are: pH 5.5, temperature 25°C, and TEMPO concentration of 10 mM. Under this optimal condition, the conversion rate of amino oxidation to aldehyde group reached more than 95%, and no degradation of the DNA backbone was detected by LC-MS.

[0127] Table 2

[0128]

[0129] Example 4: Application in DNA-encoded molecule library

[0130] The on-DNA aldehyde compound obtained in Example 1 is coupled with a target molecule (such as a small molecule drug) to react with the target molecule (such as a small molecule drug). Figure 4 , generating drug-like structures such as benzothiazole, benzimidazole, quinazoline and quinazolinone. The specific conditions are to add on-DNA aldehyde compound (4μL, 50μM dissolved in water, 0.2nmol, 1 equivalent), dimethyl sulfoxide (DMSO, 12μL), ortho-functionalized aniline (2μL, 200mM dissolved in DMSO, 400nmol, 2000 equivalents) and p-tolyl disulfide (p-tolyldisulfide, 2μL, 200mM dissolved in DMA, 400nmol, 2000 equivalents) into the reaction tube, mix well, and react at 25°C for 4 hours under 23W compact fluorescent lamp (CFL) light source.

[0131] Therefore, the method provided in the above embodiments of the present invention is suitable for constructing a high-throughput DNA encoding molecule library, and the reaction conditions are mild and highly selective.

[0132] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for synthesizing an on-DNA carbonyl compound, characterized in that: The following steps are involved: Providing an on-DNA amine compound and an on-DNA hydroxyl compound, wherein the on-DNA amine compound comprises a DNA fragment and an amine group coupled to the DNA fragment, and the on-DNA hydroxyl compound comprises a DNA fragment and a hydroxyl group coupled to the DNA fragment; The on-DNA amine compound and / or the on-DNA hydroxyl compound is reacted with laccase and an auxiliary catalyst to obtain an on-DNA carbonyl compound.

2. The synthesis method according to claim 1, characterized in that The pH value of the reaction is 4.0-7.5; and / or The reaction temperature is 25°C-37°C; and / or The reaction time is 2-48 hours.

3. The synthesis method according to claim 1, characterized in that The dosage of the laccase is 1 to 100 U / mL.

4. The synthesis method according to claim 1, characterized in that The auxiliary catalyst is selected from at least one of TEMPO, HBT, ABTS, NaClO, and NaClO2; and / or The concentration of the auxiliary catalyst is 0.01 mmol / L to 1 mol / L.

5. The synthesis method according to any one of claims 1 to 4, characterized in that The on-DNA amine compound is selected from at least one of Compound 1 to Compound 59: and / or The on-DNA hydroxy compound is selected from at least one of Compound 60 to Compound 83:

6. The synthesis method according to any one of claims 1 to 4, characterized in that The synthesis method comprises the following steps: S11, providing an on-DNA amine compound as shown in the general formula (I): S12, reacting the on-DNA amine compound in a reaction system containing laccase and an auxiliary catalyst to obtain an on-DNA carbonyl compound as shown in the general formula (II): in, is the connection key, R 1 is selected from one of hydrogen, alkyl, cycloalkyl, alkenyl and aryl, and the substituents on the aryl are independently selected from at least one of H, C1-C6 alkyl, halogen, nitro, amino, amine, alkoxy and cyano, R 2 and R 3 Each is independently selected from hydrogen, alkyl, halogenated alkyl, aryl, and other groups, wherein the other groups contain at least one of alkyl, acyl, and ester groups, and the substituents on the aryl are independently selected from at least one of H, C1-C6 alkyl, halogen, nitro, amino, amine, alkoxy, and cyano. R 4 A group selected from at least one of an alkyl group, a haloalkyl group, a cycloalkyl group, an aryl group, and a heteroaryl group, wherein the substituents on the aryl group and the heteroaryl group are independently selected from at least one of H, a C1-C6 alkyl group, a halogen group, a nitro group, an amino group, an amine group, an alkoxy group, and a cyano group; Alternatively, the synthesis method comprises the following steps: S21, providing an on-DNA hydroxy compound as shown in the general formula (III): S22, reacting the on-DNA hydroxyl compound in a reaction system containing laccase and an auxiliary catalyst to obtain an on-DNA carbonyl compound as shown in the general formula (IV): in, is the connection key; R 5 It is selected from one of aryl and heteroaryl, and the substituents on the aryl and heteroaryl are independently selected from at least one of H, C1-C6 alkyl, halogen, nitro, amino, amine, alkoxy and cyano.

7. The synthesis method according to claim 6, characterized in that The on-DNA amine compound is selected from compound 1 to compound 59; The on-DNA hydroxy compound is selected from Compound 60 to Compound 83.

8. The synthesis method according to claim 6, characterized in that The pH value of the reaction is 4.0-7.5, the temperature is 25°C-37°C, and the reaction time is 2-48 hours; and / or The reaction system contains sodium acetate buffer, and the pH value of the sodium acetate buffer is 4.0-7.

5.

9. The synthesis method according to claim 6, characterized in that The laccase is used in an amount of 1 to 100 U / mL; and / or The auxiliary catalyst is selected from at least one of TEMPO, HBT, ABTS, NaClO, and NaClO2; and / or The concentration of the auxiliary catalyst is 0.1 mmol / L to 10 mmol / L.

10. Use of the on-DNA carbonyl compound obtained by the synthesis method according to any one of claims 1 to 9 in constructing a DNA-encoded molecule library, coupling fluorescent labeling molecules, and functional modification for targeted drug delivery.

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