CN113684540A - Method for reducing On-DNA aromatic nitro compound into On-DNA aromatic amine compound - Google Patents

Method for reducing On-DNA aromatic nitro compound into On-DNA aromatic amine compound Download PDF

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CN113684540A
CN113684540A CN202010415852.2A CN202010415852A CN113684540A CN 113684540 A CN113684540 A CN 113684540A CN 202010415852 A CN202010415852 A CN 202010415852A CN 113684540 A CN113684540 A CN 113684540A
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李进
彭婷
苏立强
冯静
刘观赛
万金桥
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Abstract

The invention relates to a method for preparing an On-DNA arylamine compound by reducing nitro of the On-DNA arylamine compound into amino through reduction reaction, the method takes the On-DNA nitro compound as a substrate, and reduces the nitro compound into the On-DNA arylamine compound under the condition of thiourea dioxide and alkalinity; and can obtain highly diverse DNA-encoding compounds in a short time and at a high yield, and is suitable for synthesis of libraries of DNA-encoding compounds using multiwell plates.

Description

Method for reducing On-DNA aromatic nitro compound into On-DNA aromatic amine compound
Technical Field
The invention belongs to the technical field of coding compound libraries, and particularly relates to a method for reducing an On-DNA nitryl in the construction of a DNA coding compound library.
Background
In drug development, especially new drug development, high-throughput screening for biological targets is one of the main means for rapidly obtaining lead compounds. However, traditional high throughput screening based on single molecules requires long time, large equipment investment, limited number of library compounds (millions), and the building of compound libraries requires decades of accumulation, limiting the efficiency and possibility of discovery of lead compounds. The recent DNA-encoded compound library technologies (WO2005058479, WO2018166532, CN103882532) combine the technologies of combinatorial chemistry and molecular biology, add a DNA tag to each compound on the molecular level, and synthesize up to hundred million levels of compound libraries in a very short time, which is a trend of the next generation compound library screening technology, and begin to be widely applied in the pharmaceutical industry, resulting in many positive effects (Accounts of Chemical Research,2014,47, 1247-.
The DNA coding compound library can rapidly generate a giant compound library through combinatorial chemistry, and can screen out a lead compound with high flux, so that the screening of the lead compound becomes unprecedented rapidness and high efficiency. One of the challenges in constructing libraries of DNA-encoding compounds is the need to synthesize chemically diverse small molecules on DNA in high yields. Because DNA can be kept stable under certain conditions (solvent, pH, temperature and ion concentration), the On-DNA reaction applied to the construction of the DNA coding compound library also needs higher yield. Therefore, the reagent type, reaction type and reaction condition of the chemical reaction (On-DNA reaction for short) carried out On DNA directly influence the richness and selectivity of the DNA coding compound library. Therefore, the development of chemical reactions compatible with DNA is also a long-term research and research direction of the current DNA coding compound library technology, and the application and commercial value of the DNA coding compound library are directly influenced.
Nitro and amino groups are very common in the conversion of pharmaceutical compounds and chemically reactive functional groups, and nitro groups are often present in large numbers as a potential functional group for amino groups. The introduction of the diversity reaction of nitro group to amino group into the DNA coding compound library can further expand the diversity of the compound library, and is beneficial to improving the probability of screening effective compounds. There are several methods for converting the nitro group of On-DNA into amino group, such as the first method using hydrazine hydrate as hydrogen source and Raney Ni as reducing agent to reduce the nitro compound of On-DNA into arylamine at room temperature (ref: Bioconjugate chem.,2015, 1623-1632); the second is to reduce the On-DNA aromatic nitro compound into arylamine at 80 deg.C or lower under the condition of using water as hydrogen source, ferrous sulfate as reducing agent and sodium hydroxide (refer to ACSComb. Sci.,2018,20, 251-255); and the third is to reduce the On-DNA aromatic nitro compound into arylamine in the presence of methyl viologen, sodium hydrosulfite as reducer and methyl viologen at 80 deg.c for short time (reference: bioconjugatechem, 2017,28, 2575). The Raney Ni of the first method is not a homogeneous solution, is difficult to be added in batches by pipetting, the excessive Ni metal residue has certain destructive effect on DNA, the use of divalent Fe of the second method is a difficult problem for controlling the metal residue in a library, greenish black precipitate is easy to generate in the reaction process, the post-treatment is difficult, and the byproduct of amino sulfite is often generated in the third method and is difficult to be converted into a product.
Therefore, a novel, simple and quick nitro reduction method which does not need a metal reducing agent and is suitable for the DNA coding compound library of a large batch of porous plates is developed, the diversity of nitro reduction can be expanded, the diversity of the DNA coding compound library is increased, and the application value of the DNA coding compound library technology is improved.
Disclosure of Invention
The invention aims to provide a method for reducing an On-DNA aromatic nitro compound into an On-DNA aromatic amine compound, wherein the reaction takes the On-DNA aromatic nitro compound as a substrate, and reduces the On-DNA aromatic nitro compound into the On-DNA aromatic amine compound under the condition of thiourea dioxide and alkalinity; wherein the structural formula of the On-DNA aromatic nitro compound is DNA-Ar-NO2(ii) a The structural formula of the On-DNA arylamine compound is DNA-Ar-NH2；
Wherein, the DNA in the structural formula comprises a single-stranded or double-stranded nucleotide chain obtained by polymerizing artificially modified and/or unmodified nucleotide monomers, and the nucleotide chain is connected with Ar through one or more chemical bonds or groups;
wherein, the DNA in the structural formula is connected with Ar through one chemical bond or a plurality of chemical bonds or groups. When a chemical bond is present, the DNA in the structural formula is directly contacted with ArConnecting; multiple chemical bonds mean that the DNA and Ar in the structural formula are connected at intervals of multiple chemical bonds, for example, the DNA and Ar are connected through a methylene (-CH)2-) are linked, i.e. linked by two chemical bonds; or the DNA and the Ar are connected through a carbonyl (-CO-) to the amino of the DNA, and are also connected through two chemical bonds; or DNA and Ar through a methylene carbonyl group (-CH)2CO-) is attached to the amino group of the DNA, again by three consecutive chemical bonds.
Wherein Ar in the structural formula is selected from an optionally substituted monocyclic or bicyclic aromatic ring; -NO of the On-DNA aromatic nitro compound2-NH of said On-DNA arylamine compound attached to the ring of Ar2To the ring of Ar.
Ar is selected from the following groups:
Figure BDA0002495295260000021
R1is any one or random combination of hydrogen, halogen, cyano, hydroxyl, carboxyl, amino, substituted amino, alkyl, substituted alkyl, alkylamino, substituted alkylamino, alkoxy and substituted alkoxy, and one or more R can be arranged on Ar1The X is any one of O, S, NH or alkyl-substituted imino;
the alkyl group is C1-C12Straight or branched alkyl or C3-C8A cycloalkyl group; the alkoxy is C1-C12A straight or branched alkoxy group of (a);
the number of the substituent(s) of the substituted amino group is one or more, and the substituent(s) of the substituted amino group are independently selected from phenyl and C1-C12Alkyl radical, C3-C8One or more of cycloalkyl, alkoxyphenyl;
the number of the substituent groups of the substituted alkyl is one or more, and the substituent groups of the substituted alkyl are one or more independently selected from halogen, cyano, carboxyl, phenyl, amino and alkoxy phenyl;
the number of the substituent groups of the substituted alkylamino is one or more, and the substituent groups of the substituted alkylamino are one or more independently selected from halogen, cyano, amino, alkoxy, phenyl and alkoxyphenyl;
the number of the substituent of the substituted alkoxy is one or more, and the substituent of the substituted alkoxy is one or more independently selected from halogen, cyano, carboxyl and phenyl.
Preferably, R is1Selected from substituted alkylamino, said alkyl being C1～C6Alkyl or C3-C6A cycloalkyl group; said C is1～C6The alkyl is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl and hexyl; said C3-C6Cycloalkyl is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; the substituent of the substituted alkylamino is selected from phenyl, alkoxy or amino, and further, the alkoxy is C1～C6Alkoxy is selected from methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentoxy and hexoxy.
Preferably, R is1Is selected from substituted alkoxy, the alkoxy is C1～C6Alkoxy, specifically selected from methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentoxy, hexoxy; the substituent of the substituted alkoxy is selected from carboxyl.
Preferably, R is1Selected from substituted alkyl; the alkyl group is C1～C6An alkyl group; said C is1～C6The alkyl is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl and hexyl; the substituent of the substituted alkyl group is selected from carboxyl.
Preferably, R is1Is selected from substituted amino, and the substituent of the substituted amino is selected from phenyl, alkoxy phenyl and C1-C6Alkyl or C3-C6A cycloalkyl group; further, the saidAlkoxyphenyl is C1～C6Alkoxyphenyl, specifically selected from methoxyphenyl, ethoxyphenyl, n-propoxyphenyl, isopropoxyphenyl, n-butoxyphenyl, isobutoxyphenyl, tert-butoxyphenyl, pentoxyphenyl, and hexoxyphenyl; said C1～C6The alkyl is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl and hexyl; said C3-C6The cycloalkyl group is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
Preferably, Ar is selected from
Figure BDA0002495295260000031
Preferably, the On-DNA aromatic nitro compound is selected from:
Figure BDA0002495295260000041
the R islIs hydrogen, halogen, cyano, bayer group, carboxyl, amino, Cl-C6Alkyl radical, Cl-C6Alkylamino radical, Cl-C6A random combination of any one or more of alkoxy groups;
or said R1Selected from substituted alkylamino, said alkyl being C1～C6Alkyl or C3-C6A cycloalkyl group; said C is1～C6The alkyl is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl and hexyl; said C3-C6Cycloalkyl is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; the substituent of the substituted alkylamino is selected from phenyl, alkoxy or amino, and further, the alkoxy is C1～C6Alkoxy is selected from methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentoxy and hexoxy.
Or said R1Is selected from the group consisting ofAlkoxy, the alkoxy is C1～C6Alkoxy, specifically selected from methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentoxy, hexoxy; the substituent of the substituted alkyloxy is selected from carboxyl.
Or said R1Selected from substituted alkyl; the alkyl group is C1～C6An alkyl group; said C is1～C6The alkyl is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl and hexyl; the substituent of the substituted alkyl group is selected from carboxyl.
Or said R1Is selected from substituted amino, and the substituent of the substituted amino is selected from phenyl, alkoxy phenyl and C1-C6Alkyl or C3-C6A cycloalkyl group; further, the alkoxyphenyl group is C1～C6Alkoxyphenyl, specifically selected from methoxyphenyl, ethoxyphenyl, n-propoxyphenyl, isopropoxyphenyl, n-butoxyphenyl, isobutoxyphenyl, tert-butoxyphenyl, pentoxyphenyl, and hexoxyphenyl; said C1～C6The alkyl is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl and hexyl; said C3-C6The cycloalkyl group is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
Further, the On-DNA aromatic nitro compound is specifically selected from:
Figure BDA0002495295260000051
Figure BDA0002495295260000052
Figure BDA0002495295260000061
a method for reducing an On-DNA aromatic nitro compound into an On-DNA aromatic amine compound comprises the following steps: adding 5-200 times of molar equivalent of thiourea dioxide and 5-500 times of molar equivalent of alkali into an On-DNA aromatic nitro compound solution with the molar equivalent of 1 and the molar concentration of 0.1-2 mM, and reacting at 20-100 ℃ for 1-24 hours until the reaction is finished.
Preferably, the base is sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, cesium carbonate, N-diisopropylethylamine or 1,1,3, 3-tetramethylguanidine.
Preferably, the solution is one or a mixture of acetonitrile, methanol, ethanol, DMF, DMA, DMSO, THF, water, inorganic salt buffer solution, organic acid buffer solution and organic base buffer solution.
Preferably, the reaction is carried out in a buffer system, further, the solution is a borate buffer, and the pH of the borate buffer is 9.4.
The reaction temperature is 20 ℃, 30 ℃, 40 ℃, 50 ℃, 60 ℃, 70 ℃, 80 ℃ or 90 ℃.
The On-DNA arylnitro compound solution has a molarity of 0.1mM, 0.2mM, 0.3mM, 0.4mM, 0.5mM, 0.6mM, 0.7mM, 0.8mM, 0.9mM, 1.0mM, 1.1mM, 1.2mM, 1.3mM, 1.4mM, 1.5mM, 1.6mM, 1.7mM, 1.8mM, 1.9mM, or 2.0 mM.
Preferably, when the molar equivalent of the On-DNA arylnitro compound is 1, the molar equivalent of the thiourea dioxide compound is 10 equivalents, 30 equivalents, 50 equivalents, 70 equivalents, 90 equivalents, 100 equivalents or 200 equivalents, and the molar equivalent of the base is 50 equivalents, 100 equivalents, 200 equivalents, 300 equivalents, 400 equivalents or 500 equivalents.
Further, the reaction is fed in the order of mixing the alkali and thiourea dioxide first and then adding the mixture to the solution of the On-DNA nitro compound.
Further, the above method is used for batch multi-well plate operations.
Further, the above method is used for the synthesis of libraries of DNA-encoding compounds for multi-well plates.
The method can realize the method for reducing the nitro group in the DNA coding compound library into the amino group, and can introduce the amino group in a large scale. The method has the advantages of high yield, single product, simple post-treatment, environmental protection, suitability for large-scale multi-plate operation and provision of a new framework for a DNA coding compound library.
In the preferred embodiment of the invention, the reaction yield is improved and the accuracy of the method applied to the library construction of the DNA coding compound is improved by controlling the proportion of reaction raw materials and reagents and the feeding sequence.
Definitions of terms used in connection with the present invention: the initial definitions provided herein for a group or term apply to that group or term throughout the specification unless otherwise indicated; for terms not specifically defined herein, the meanings that would be given to them by a person skilled in the art are to be given in light of the disclosure and the context.
"substituted" means that a hydrogen atom in a molecule is replaced by a different atom or molecule.
The minimum and maximum values of the carbon atom content in the hydrocarbon group are indicated by a prefix, e.g. prefix (Ca-C)b) Alkyl means any alkyl group containing from "a" to "b" carbon atoms. Thus, for example, C1～C12The alkyl group is a straight-chain or branched alkyl group having 1 to 12 carbon atoms.
Alkyl is a straight or branched chain hydrocarbon radical derived from an alkane molecule by the removal of one hydrogen atom, e.g. methyl-CH3ethyl-CH2CH3(ii) a The alkyl group may also be part of other groups such as C1-C6 alkoxy, C1-C6 alkylamino.
Cycloalkyl groups: refers to saturated or partially saturated cyclic groups having multiple carbon atoms and no ring heteroatoms and having a single ring or multiple rings (including fused, bridged, and spiro ring systems).
The halogen is fluorine, chlorine, bromine or iodine.
Alkoxy groups: means that the alkyl radical is bound to an oxygen atom to form a substituent, e.g. methoxy is-OCH3。
Alkylamino group: meaning that the alkyl radical is linked to the amino nitrogen atom to form a substituent, e.g. methylamino is-NH2CH3。
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples, but it should not be construed that the scope of the above subject matter is limited to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Drawings
FIG. 1: the corresponding conversion rate distribution diagram of the 29 On-DNA arylamine compounds synthesized in the embodiment 4 of the invention.
Detailed Description
The raw materials and equipment used in the invention are known products and are obtained by purchasing commercial products.
In the invention, the term "room temperature" means 20-25 ℃.
DMA: dimethylacetamide (Dimethylacetamide); DMF: dimethylformamide (DMF).
HATU: 2- (7-azabenzotriazole) -N, N' -tetramethyluronium hexafluorophosphate;
DIPEA: n, N-diisopropylethylamine;
DMSO, DMSO: dimethyl sulfoxide; THF: tetrahydrofuran.
DNA-NH in the present invention2Or
Figure BDA0002495295260000081
DNA constructs with a-NH 2 linker formed by single-or double-stranded DNA and linker groups, e.g.DNA-NH of "compound 1" in WO20050584792And (5) structure.
Example 1 reduction of On-DNA Nitro Compounds to On-DNA arylamine Compounds
1) Synthesis of On-DNA nitro compound
Figure BDA0002495295260000082
Reacting DNA-NH2Dissolved in borate bufferA solution (0.25M, pH 9.4) was prepared as a 1mM concentration solution (10 μ L, 10nmol), to which a premixed solution (an aromatic nitro compound containing a carboxylic acid (100 molar equivalent, 200mM DMA solution), HATU (100 molar equivalent, 200mM DMA solution), DIPEA (100 molar equivalent, 200mM DMA solution) was added, reacted at room temperature for 3 hours, and after completion of the reaction, ethanol precipitation treatment was performed to obtain an On-DNA nitro compound intermediate, which was used directly in the next step;
the On-DNA nitro compound 2 generated in situ was dissolved in borate buffer (0.25M, pH 9.4) to prepare a 1mM concentration solution (10 μ L, 10nmol), phenethylamine (200 molar equivalent, 200mM DMSO solution) was added to the solution to react at 75 ℃ for 16 hours, and after completion of the reaction, ethanol precipitation treatment was performed to obtain On-DNA nitro compound 3, which was used directly in the next step.
2) Synthesis of On-DNA arylamine compounds
Figure BDA0002495295260000083
The On-DNA arylnitro compound 3 was dissolved in borate buffer (0.25M, pH 9.4) to prepare a 1mM concentration solution (10 μ L, 10nmol), and the solution was added with a preliminary mixture (NaOH (100 molar equivalent, 1000mM aqueous solution), thiourea dioxide (50 molar equivalent, 200mM aqueous solution)) and reacted at room temperature for 2 hours;
and (3) after the reaction is finished, carrying out ethanol precipitation: adding a 5M sodium chloride solution with the total volume of 10% into the solution, then continuously adding anhydrous ethanol with the volume of 3 times of the total volume, uniformly oscillating, freezing the reaction at-20 ℃ for 2 hours, centrifuging at 12000rpm for half an hour, pouring out the supernatant, and carrying out vacuum freeze-drying On the rest precipitate at-40 ℃ to obtain an On-DNA arylamine compound 4, and sending LCMS to confirm that the reaction conversion rate is 90%.
Example 2 reduction of On-DNA Nitro Compounds to On-DNA arylamine Compounds
Figure BDA0002495295260000091
Dissolving the On-DNA aromatic nitro compound 3 in a solvent X to prepare a 1mM concentration solution (10 mu L, 10nmol), mixing NaOH (Z molar equivalent, 1000mM aqueous solution) and thiourea dioxide (Y molar equivalent, 200mM aqueous solution) for 5min at room temperature, adding the mixed solution into the On-DNA aromatic nitro compound 3 solution, and reacting for 2 hours at room temperature;
and (3) after the reaction is finished, carrying out ethanol precipitation: adding a 5M sodium chloride solution with the total volume of 10% into the solution, then continuously adding anhydrous ethanol with the volume of 3 times of the total volume, uniformly oscillating, freezing the reaction at-20 ℃ for 2 hours, centrifuging at 12000rpm for half an hour, pouring out the supernatant, and carrying out vacuum freeze-drying On the rest precipitate at-40 ℃ to obtain an On-DNA arylamine compound 4, and sending to LCMS to confirm the conversion rate of the reaction. Specific reaction conditions are shown in table 1 below.
The solvent is selected from water or borate buffer (0.25M, pH 9.4).
Table 1: example 2 specific reaction conditions and yield data Table
Figure BDA0002495295260000092
Figure BDA0002495295260000101
The above experimental results show that the yield is highest under the reaction conditions of experiment 2.
Example 3 reduction of On-DNA Nitro Compounds to On-DNA arylamine Compounds
Figure BDA0002495295260000102
On-DNA arylnitro compound 3 was dissolved in borate buffer (0.25M, pH 9.4) to prepare a 1mM concentration solution (10 μ L, 10nmol), thiourea dioxide (50 molar equivalent, 200mM aqueous solution), NaOH (100 molar equivalent, 1000mM aqueous solution) was added to the solution in this order, and reacted at room temperature for 2 hours in one pot;
and (3) after the reaction is finished, carrying out ethanol precipitation: adding a 5M sodium chloride solution with the total volume of 10% into the solution, then continuously adding anhydrous ethanol with the volume of 3 times of the total volume, uniformly oscillating, freezing the reaction at-20 ℃ for 2 hours, centrifuging at 12000rpm for half an hour, pouring out the supernatant, and carrying out vacuum freeze-drying On the rest precipitate at-40 ℃ to obtain an On-DNA arylamine compound 4, and sending LCMS to confirm that the reaction conversion rate is 78%.
Example 4 reduction of On-DNA Nitro Compounds to On-DNA arylamine Compounds
DNA-NH2→DNA-NH-Ar-NO2
Reacting DNA-NH2Dissolved in borate buffer (0.25M, pH 9.4) to prepare a 1mM concentration solution (10. mu.L, 10nmol), to which an arylnitro compound (50 molar equivalents, 200mM DMSO solution) was added for reaction at 90 ℃ for 16 hours, and after completion of the reaction, ethanol precipitation treatment was performed to obtain an On-DNA arylnitro compound, which was used directly in the next step.
29 On-DNA aromatic nitro compounds were synthesized according to the above method.
1) Synthesis of On-DNA arylamine compounds
Figure BDA0002495295260000104
29 On-DNA aromatic nitro compounds were dissolved in borate buffer (0.25M, pH 9.4) to prepare 1mM concentration solutions (10 μ L, 10nmol), NaOH (100 equivalents, 1000mM aqueous solution), thiourea dioxide (50 equivalents, 200mM aqueous solution) were mixed at room temperature for 5min, and then the mixture was added to 29 On-DNA aromatic nitro compound solutions, respectively, and reacted at room temperature for 2 hours;
and (3) after the reaction is finished, carrying out ethanol precipitation: adding a 5M sodium chloride solution with the total volume of 10% into the solution, then continuously adding anhydrous ethanol with the volume of 3 times of the total volume, uniformly oscillating, freezing the reaction at-20 ℃ for 2 hours, centrifuging at 12000rpm for half an hour, pouring out the supernatant, and carrying out vacuum freeze-drying On the rest precipitate at-40 ℃ to obtain an On-DNA arylamine compound, sending to LCMS to confirm the reaction conversion rate, wherein the specific compound structure and the yield are shown in figure 1.
In conclusion, the method can obtain the On-DNA amino compound from the On-DNA nitro compound under thiourea oxide and alkaline conditions by controlling the conditions of the feeding sequence, the temperature, the pH value and the like during the reaction. The method has the advantages of wide substrate application range, simple post-treatment, environmental friendliness, capability of obtaining a highly diversified DNA coding compound library in a short time at a high yield, and suitability for operation by using a large batch of multi-well plates.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.

Claims (11)

1. A method for reducing an On-DNA aromatic nitro compound into an On-DNA aromatic amine compound is characterized in that: the method takes an On-DNA aromatic nitro compound as a substrate, and reduces the On-DNA aromatic nitro compound into an On-DNA aromatic amine compound under the condition of thiourea dioxide and alkalinity; wherein the structural formula of the On-DNA aromatic nitro compound is DNA-Ar-NO2(ii) a The structural formula of the On-DNA arylamine compound is DNA-Ar-NH2；
Wherein the DNA in the structural formula comprises a single-stranded or double-stranded nucleotide chain obtained by polymerizing artificially modified and/or unmodified nucleotide monomers, and the nucleotide chain is connected with Ar through one or more chemical bonds or groups;
wherein Ar in the structural formula is selected from an optionally substituted monocyclic or bicyclic aromatic ring; -NO of the On-DNA aromatic nitro compound2-NH of said On-DNA arylamine compound attached to the ring of Ar2To the ring of Ar.
2. The method of claim 1, wherein: ar is selected from the following groups:
Figure FDA0002495295250000011
R1is any one or random combination of hydrogen, halogen, cyano, hydroxyl, carboxyl, amino, substituted amino, alkyl, substituted alkyl, alkylamino, substituted alkylamino, alkoxy and substituted alkoxy, and one or more R can be arranged on Ar1The X is any one of O, S, NH or alkyl-substituted imino;
the alkyl group is C1-C12Straight or branched alkyl or C3-C8A cycloalkyl group; the alkoxy is C1-C12A straight or branched alkoxy group of (a);
the number of the substituent(s) of the substituted amino group is one or more, and the substituent(s) of the substituted amino group are independently selected from phenyl and C1-C12Alkyl radical, C3-C8One or more of cycloalkyl, alkoxyphenyl;
the number of the substituent groups of the substituted alkyl is one or more, and the substituent groups of the substituted alkyl are one or more independently selected from halogen, cyano, carboxyl, phenyl, amino and alkoxy phenyl;
the number of the substituent groups of the substituted alkylamino is one or more, and the substituent groups of the substituted alkylamino are one or more independently selected from halogen, cyano, amino, alkoxy, phenyl and alkoxyphenyl;
the number of the substituent of the substituted alkoxy is one or more, and the substituent of the substituted alkoxy is one or more independently selected from halogen, cyano, carboxyl and phenyl.
3. The method of claim 1, wherein: adding 5-200 times of molar equivalent of thiourea dioxide and 5-500 times of molar equivalent of alkali into an On-DNA aromatic nitro compound solution with the molar equivalent of 1 and the molar concentration of 0.1-2 mM, and reacting at 20-100 ℃ for 1-24 hours until the reaction is finished.
4. The method of claim 3, wherein: the base is selected from sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, cesium carbonate, N-diisopropylethylamine or 1,1,3, 3-tetramethylguanidine.
5. The method of claim 3, wherein: the solution is one or a mixture of acetonitrile, methanol, ethanol, DMF, DMA, DMSO, THF, water, inorganic salt buffer solution, organic acid buffer solution and organic base buffer solution.
6. The method of claim 3, wherein: the reaction temperature is 20 ℃, 30 ℃, 40 ℃, 50 ℃, 60 ℃, 70 ℃, 80 ℃ or 90 ℃.
7. The method of claim 3, wherein: the On-DNA arylnitro compound solution has a molarity of 0.1mM, 0.2mM, 0.3mM, 0.4mM, 0.5mM, 0.6mM, 0.7mM, 0.8mM, 0.9mM, 1.0mM, 1.1mM, 1.2mM, 1.3mM, 1.4mM, 1.5mM, 1.6mM, 1.7mM, 1.8mM, 1.9mM, or 2.0 mM.
8. The method of claim 3, wherein: when the molar equivalent of the On-DNA aromatic nitro compound is 1, the molar equivalent of thiourea dioxide is 10 equivalents, 30 equivalents, 50 equivalents, 70 equivalents, 90 equivalents, 100 equivalents or 200 equivalents, and the molar equivalent of the base is 50 equivalents, 100 equivalents, 200 equivalents, 300 equivalents, 400 equivalents or 500 equivalents.
9. The method of claim 3, wherein: the charging sequence of the reaction is that firstly, alkali and thiourea dioxide are mixed and then added into the solution of the On-DNA nitro compound.
10. The method according to any one of claims 1-9, wherein: the method is used for batch multi-well plate operations.
11. The method according to any one of claims 1-9, wherein: the method is used for the synthesis of libraries of DNA-encoding compounds for multi-well plates.
CN202010415852.2A 2020-05-18 2020-05-18 Method for reducing On-DNA aromatic nitro compound into On-DNA aromatic amine compound Pending CN113684540A (en)

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Citations (1)

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Publication number Priority date Publication date Assignee Title
CN109680342A (en) * 2018-12-18 2019-04-26 上海药明康德新药开发有限公司 The method that On-DNA virtue nitro compound is reduced into On-DNA aromatic amine compound in DNA encoding compound library

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CN109680342A (en) * 2018-12-18 2019-04-26 上海药明康德新药开发有限公司 The method that On-DNA virtue nitro compound is reduced into On-DNA aromatic amine compound in DNA encoding compound library

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