CN114075257B - Method for preparing arylamine compound from On-DNA aryl halide - Google Patents
Method for preparing arylamine compound from On-DNA aryl halide Download PDFInfo
- Publication number
- CN114075257B CN114075257B CN202010800122.4A CN202010800122A CN114075257B CN 114075257 B CN114075257 B CN 114075257B CN 202010800122 A CN202010800122 A CN 202010800122A CN 114075257 B CN114075257 B CN 114075257B
- Authority
- CN
- China
- Prior art keywords
- dna
- aryl
- solution
- alkyl
- compound
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
- C07H21/04—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
-
- C—CHEMISTRY; METALLURGY
- C40—COMBINATORIAL TECHNOLOGY
- C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
- C40B40/00—Libraries per se, e.g. arrays, mixtures
- C40B40/04—Libraries containing only organic compounds
- C40B40/06—Libraries containing nucleotides or polynucleotides, or derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C40—COMBINATORIAL TECHNOLOGY
- C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
- C40B50/00—Methods of creating libraries, e.g. combinatorial synthesis
- C40B50/08—Liquid phase synthesis, i.e. wherein all library building blocks are in liquid phase or in solution during library creation; Particular methods of cleavage from the liquid support
Abstract
The invention relates to a method for preparing arylamine compounds from On-DNA aryl halides, which takes On-DNA aryl halides as substrates, reacts with various amine compounds under the action of palladium catalysts, ligands and alkali, and is converted into On-DNA arylamine compounds. The method has the advantages of wide substrate range, friendly reaction environment, high raw material conversion rate, small damage to DNA molecules, simple post-treatment and the like, and is suitable for synthesizing the DNA coding compound library by using a porous plate.
Description
Technical Field
The invention belongs to the technical field of coded compound libraries, and particularly relates to a method for converting On-DNA aryl halides into arylamine compounds in construction of a DNA coded compound library.
Background
In drug development, especially new drug development, high throughput screening against biological targets is one of the main means to rapidly obtain lead compounds. However, conventional high throughput screening based on single molecules requires long time, huge equipment investment, limited numbers of library compounds (millions), and the build-up of compound libraries requires decades of accumulation, limiting the efficiency and possibilities of discovery of lead compounds. The DNA-encoded compound libraries technology (WO 2005058479, WO2018166532, CN 103882532) which has emerged in recent years, combined with combinatorial chemistry and molecular biology techniques, allows up to billions of compound libraries to be synthesized in very short time by tagging each compound with a DNA tag at the molecular level. And the compound can be identified by a gene sequencing method, so that the size and the synthesis efficiency of a compound library are greatly increased, the compound library becomes a trend of the next generation of compound library screening technology, and the compound can be widely applied to the foreign pharmaceutical industry, and a plurality of positive effects are produced (Accounts of Chemical Research,2014,47,1247-1255).
The DNA encoding compound library rapidly generates a huge compound library by combinatorial chemistry, and can screen the lead compound with high flux, so that the screening of the lead compound becomes unprecedented rapid and efficient. In the development of DNA coding molecule library drug technology, the synthesis technology and the screening technology of the DNA coding molecule library are two key points, and often determine the drug screening effect. The kind of the reagent, the kind of the reaction and the reaction condition of the chemical reaction (called On-DNA reaction for short) performed On the DNA directly influence the richness and the selectivity of the DNA coding molecule library.
Aromatic compounds are an important component of organic compounds, which are widely found in some natural products and bioactive molecules. Meanwhile, aromatic hydrocarbon compounds are widely applied to chemical fields such as pesticides, medicines, chemical industry, dyes and the like as cheap chemical raw materials. Among them, benzyl-substituted backbone compounds are often contained in natural products, and are therefore of particular importance in connection with the synthesis of such molecules.
The On-DNA reaction requires that the stability of DNA be maintained under certain reaction conditions (solvent, pH, temperature and ion concentration), and the reaction applied to the construction of DNA coding compound library also requires higher yield, while the application of the currently published On-DNA reaction to DNA coding compound library technology is greatly limited by dozens of On-DNA reactions, and the method for constructing On-DNA arylamine compounds by On-DNA aryl halogen compounds has been reported (1. Med. Chem. Commun.,2017,8,1614-1617;2.Chem. Eur. J.,2018,24 (8), 1795-1800;3.Med. Chem. Commun.,2018,9,1188). This patent adopts a new reaction system, has improved the application range of arylamine.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a method for converting On-DNA aryl halide into arylamine compound, which uses On-DNA aryl halide as substrate, reacts with different kinds of amine compound under the action of alkali, palladium catalyst and ligand, and converts On-DNA arylamine compound. The method has the advantages of mild reaction conditions, high selectivity, high yield and simple post-treatment, is suitable for producing DNA coding compound libraries, and can remarkably improve the diversity of molecules in the compound libraries.
In order to solve the technical problems, the invention adopts the following technical scheme:
aryl group formed by On-DNAThe method for preparing the arylamine compound from the halide comprises the steps of taking an On-DNA aryl halide compound as a substrate, and reacting with the amine compound under the action of alkali, palladium catalyst and ligand to generate the On-DNA arylamine compound; the structural formula of the On-DNA aryl halogenated compound is DNA-Ar-X, and the structural formula of the amine compound is as follows:the structural formula of the On-DNA arylamine compound is->
Wherein the DNA in the formula comprises a single-stranded or double-stranded nucleotide chain polymerized from artificially modified and/or unmodified nucleotide monomers, the nucleotide chain being linked to Ar by one or more chemical bonds or groups; the length of the single-stranded or double-stranded nucleotide chain is 5-200 bp; -X of said On-DNA aryl halide is attached to the ring of Ar, X being selected from chlorine, bromine or iodine; the size of the DNA is 10 to 200 bases.
Wherein Ar in the structural formula is selected from a substituted aromatic ring or aromatic heterocyclic ring; r is R 1 、R 2 Selected from hydrogen or a group having a molecular weight of 1000 or less which is directly bonded to the amino nitrogen atom.
Preferably, ar is selected from:
wherein Ar has one or more substituents, the substituents are any one or more of hydrogen, halogen, carboxyl, cyano, hydroxyl, alkyl, substituted alkyl, alkoxy and substituted alkoxy, and the alkyl is C 1 -C 20 Straight or branched alkyl of (a); the alkoxy is C 1 -C 20 Straight or branched alkoxy of (a);
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 selected from halogen, cyano and carboxyl independently;
the number of the substituent groups of the substituted alkoxy is one or more, and the substituent groups of the substituted alkoxy are one or more selected from halogen, cyano and carboxyl independently.
As preferable: said R is 1 、R 2 Selected from hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, aromatic heterocyclic group or substituted aromatic heterocyclic group; wherein the alkyl group is C 1 ～C 20 An alkyl group; the number of substituents for the substituted alkyl group is one or more; the substituent of the substituted alkyl is one or more of halogen, nitro, alkoxy, halogenated phenyl, phenyl and alkylphenyl which are independent of each other;
the aryl is selected from phenyl or naphthyl; the number of the substituent groups of the substituted aryl is one or more, and the substituent groups of the substituted aryl are one or more selected from halogen, cyano, nitro, carboxyl, alkoxy, halogen substituted alkoxy, alkyl, halogen substituted alkyl, aromatic heterocyclic group, alkyl substituted aromatic heterocyclic group and amine acyl independently;
the aromatic heterocyclic group refers to 5-10 single cyclic groups or multiple cyclic groups containing C, O, S, N and other atoms with aromaticity; the number of the substituent groups of the substituted aromatic heterocyclic group is one or more, and the substituent groups of the substituted aromatic heterocyclic group are one or more selected from halogen, cyano, nitro, alkyl, halogen substituted alkyl, cycloalkyl, alkoxy, halogen substituted alkoxy, phenyl, halogenated phenyl, alkylphenyl, heterocyclic group, alkanoyl, carbonyl, sulfonyl and alkylsulfonyl which are independent of each other;
or R is 1 、R 2 And are connected to form 3-8 membered saturated heterocycle.
As preferable: the On-DNA aryl halogenated compound is specifically selected from the following structures:
as preferable: said R is 1 、R 2 Selected from hydrogen, C 1 ～C 6 Alkyl or aromatic heterocyclic group, wherein the aromatic heterocyclic group is pyridyl, quinolyl, isoquinolyl or pyrimidinyl; the C is 1 ～C 6 The alkyl is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl and hexyl.
As preferable: said R is 1 、R 2 Selected from hydrogen and substituted aromatic heterocyclic groups, wherein the aromatic heterocyclic groups are single cyclic groups or multiple cyclic groups with aromaticity formed by 5-10 atoms such as C, O, S, N and the like; the number of the substituent groups of the substituted aromatic heterocyclic group is one or more, and the substituent groups of the substituted aromatic heterocyclic group are mutually independent cyano groups, halogen and C 1 ～C 6 Alkyl, halogenated C 1 ～C 6 Alkyl, C 1 ～C 6 Alkoxy, halo C 1 ～C 6 Alkoxy, saturated nitrogen-containing five-membered heterocyclic group, phenyl, halogenated phenyl and saturated C 3 ～C 6 Cycloalkyl, carbonyl, sulfonyl, C 1 ～C 6 Alkylsulfonyl, C 1 ～C 6 One or more of alkanoyl groups; the C is 1 ～C 6 Alkyl is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, and hexane; the C is 1 ～C 6 Alkoxy is selected from methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy; the saturated C 3 ～C 6 Cycloalkyl is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl; the C is 1 ～C 6 Alkanoyl is selected from the group consisting of methanoyl, ethaneacyl, n-propanoyl, isopropanoyl, n-butanoyl, isobutane acyl, t-butanoyl, pentanoyl, hexanoyl; the halogen is fluorine, chlorine or bromine.
As preferable: said R is 1 、R 2 Selected from hydrogen, C 1 ～C 6 Alkyl, substituted aryl; the aryl is selected from phenyl or naphthyl; the number of substituents of the substituted aryl is one or more, and the substituents of the substituted aryl are independently selected from halogen, cyano, nitro, carboxyl and C 1 ～C 6 Alkoxy, halogen substituted alkoxy, alkyl, halogen substituted alkyl, nitrogen containing five membered aromatic heterocyclic group, C 1 ～C 6 One or more of alkyl substituted nitrogen-containing five-membered aromatic heterocyclic group and aminoacyl; the C is 1 ～C 6 Alkyl is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, and hexane; the C is 1 ～C 6 Alkoxy is selected from methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy; the halogen is fluorine, chlorine or bromine; the aminoacyl is selected from methylaminoyl, ethylamino, n-alanyl, isopropylamino, n-butyryl, isobutylaminoyl, t-butyrylaminoyl, valerylaminoyl, caproaminoyl and cyclohexaminoyl;
as preferable: said R is 1 、R 2 Selected from hydrogen, C 1 ～C 6 Alkyl, aryl; the aryl is selected from phenyl or naphthyl; the C is 1 ～C 6 The alkyl is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl and hexyl.
The amine compound is specifically selected from the following structures:
the invention provides a method for preparing arylamine compounds by On-DNA aryl halides, which comprises the following steps: adding alkali, amine compound, palladium catalyst and ligand into aryl halogenated compound solution with molar equivalent of 1 and molar concentration of 0.5-5mM, and reacting at 20-100 ℃ for 0.5-24 hours until the reaction is finished to generate On-DNA aromatic amine compound.
The chemical reaction equation of the above reaction is:
further, the base in the process is selected from sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, cesium carbonate, N-diisopropylethylamine or 1, 3-tetramethylguanidine;
the palladium catalyst is [ (pi-all) PdCl] 2 (allylpalladium chloride dimer); the ligand is as follows: cBIDP with the structural formula of
Further, the reaction is carried out in a solvent, wherein the solvent is one or a mixed solvent of a plurality of acetonitrile, methanol, ethanol, DMF, DMA, DMSO, THF, water, inorganic salt buffer solution, organic acid buffer solution and organic base buffer solution; preferably, the solvent is a mixed solution of water and DMA, and the DMA accounts for 10-60% of the volume of the solution. More preferably, DMA is 40% to 60% of the solution volume.
Further, the reaction temperature is 20-100 ℃; preferably, the reaction temperature is 40-90 ℃; more preferably, the reaction temperature is 50 ℃, 60 ℃, 70 ℃ or 80 ℃;
further, the reaction time is 0.5 to 24 hours; preferably, the reaction time is 0.5 to 16 hours; more preferably, the reaction time is 1 hour, 2 hours, 4 hours.
Further, the molar equivalent of DNA-Ar-X in the reaction is 1, the molar equivalent of the amine compound is 5-1000, and the molar equivalent of the alkali is 5-1000; the molar equivalent of the palladium catalyst is 0.5-100; the molar equivalent of the ligand is 1-200.
Preferably, the molar equivalent of the amine compound is 300-1000; preferably, the molar equivalent of the amine compound is 400, 500, 600, 700, 800 or 900.
Preferably, the molar equivalent of the base is 200-1000; more preferably, the molar equivalent of the base is 300, 400, 500, 600, 700 or 800.
Preferably, the molar equivalent of the palladium catalyst is 0.5-5; more preferably, the molar equivalent of the palladium catalyst is 0.6, 0.8, 1.0, 1.2, 1.5, 2.0 or 3.0.
Preferably, the molar equivalent of the ligand is 1-10; more preferably, the ligand has a molar equivalent of 1.5, 2.0, 3.0, 5.0 or 8.0.
Further, the reaction reagent is added in the order of adding aryl halide compound, alkali and amine compound, adding palladium catalyst and adding ligand.
Further, the method is used for batch multi-well plate operations.
Further, the method is used for the synthesis of DNA encoding compound libraries in multiwell plates.
The method can realize the conversion from On-DNA aryl halogenated compounds to arylamine compounds in the construction of DNA coding compound library. The method has high yield and single product, can be carried out in a mixed water phase of an organic solvent/water phase, is simple to operate, and is suitable for synthesizing the DNA coding compound library by using a porous plate.
In the preferred embodiment of the invention, the generation of byproducts can be inhibited by controlling the proportion of the solvent of the reaction system, the reaction temperature and the material ratio, so that the accuracy of the method applied to the library construction of the DNA coding compound library is improved.
Definition of terms used in connection with the present invention: unless otherwise indicated, the initial definitions provided for groups or terms herein apply to the groups or terms throughout the specification; for terms not specifically defined herein, the meanings that one skilled in the art can impart based on 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 represented by prefixes, for example, prefixes (Ca to C b ) Alkyl indicates any alkyl group containing from "a" to "b" carbon atoms. Thus, for example, C 1 ～C 20 Alkyl refers to straight or branched chain alkyl groups containing 1 to 20 carbon atoms.
Alkyl refers to a straight or branched hydrocarbon radical containing at least one hydrogen atom in the alkane molecule, e.g. methyl-CH 3 ethyl-CH 2 CH 3 The method comprises the steps of carrying out a first treatment on the surface of the The alkyl group may also be part of another group, such as C 1 ～C 6 An alkoxy group.
Cycloalkyl: 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.
Haloalkyl: means that hydrogen atoms on the alkyl group are partially or completely substituted with halogen atoms.
An alkoxy group: refers to alkyl groups bound to oxygen atoms to form substituents, e.g. methoxy groups of-OCH 3 。
Halogenated phenyl: represents halogen-substituted phenyl.
Alkylphenyl: represents an alkyl-substituted phenyl group.
A heterocyclic group: is a saturated or unsaturated, monocyclic or polycyclic hydrocarbon group of 3 to 8 atoms which carries at least one atom selected from O, S, N.
Aryl: refers to an aromatic single cyclic or multiple cyclic groups composed of C atoms without heteroatoms.
An aromatic heterocyclic group refers to a single ring or a fused multiple ring having a conjugated pi electron system comprising one to more heteroatoms, containing at least one ring heteroatom selected from N, O or S, the remaining ring atoms being C, and further having a fully conjugated pi electron system. Such as furan, pyrrole, quinoline, thiophene, pyridine, pyrazole, N-alkylpyrrole, pyrimidine, pyrazine, imidazole, tetrazole, thienopyridinyl, and the like.
It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.
Drawings
Fig. 1 to 4: 71 On-DNA arylamine compounds obtained in example 4 of the present invention and the corresponding conversion rates.
Fig. 5 to 8: 59 On-DNA arylamine compounds obtained in example 5 of the present invention and the corresponding conversion rates.
Detailed Description
The raw materials and equipment used in the invention are all known products and are obtained by purchasing commercial products.
DMA: n, N-dimethylacetamide, DMF: dimethylformamide.
DMSO: dimethyl sulfoxide; THF: tetrahydrofuran.
HATU:2- (7-azabenzotriazol) -N, N' -tetramethylurea hexafluorophosphate;
DIPEA: n, N-diisopropylethylamine.
DNA-NH in the present invention 2 Is formed by single-stranded or double-stranded DNA and a linker group and carries-NH 2 DNA structure of linker, e.g. DNA-NH of "component 1" in WO2005058479 2 Structure is as follows. Also for example the following DNA structure:
wherein A is adenine, T is thymine, C is cytosine, and G is guanine.
EXAMPLE 1 Synthesis of On-DNA arylamine Compounds
The synthesis of the On-DNA aryl iodide comprises the following specific steps:
aromatic group is added at 4 DEG CThe baseband (50 molar equivalents, 200mM DMA solution) was mixed with HATU (50 molar equivalents, 200mM DMA solution), DIPEA (50 molar equivalents, 200mM DMA solution) and left for 10min; subsequent reaction to DNA-NH 2 To the solution (1 molar equivalent, 1mM aqueous solution) was added the above mixture, and the mixture was stirred and reacted at 25℃for 2 hours. Ethanol precipitation is carried out after the reaction is finished: adding 10% volume of 5M sodium chloride aqueous solution into the solution, then continuously adding 3 times volume of cold ethanol, standing for 20 minutes at-20 ℃ after shaking uniformly, centrifuging at 12000 speed for 5 minutes, pouring out supernatant, and dissolving the rest precipitate with deionized water to obtain the solution of the On-DNA aryl iodide.
The On-DNA aryl iodide reacts with aniline under the catalysis of palladium to generate an On-DNA arylamine compound, and the specific reaction steps are as follows:
all solvents were sonicated for 30 min, to On-DNA aryl iodide solution (1 molar equivalent, 1mM in water), aqueous sodium hydroxide solution (300 molar equivalent, 1M in water), DMA solution of aniline (500 molar equivalent, 1M in DMA solution), DMA solution of palladium catalyst (0.6 molar equivalent, 12.5mM in DMA solution) and ligand (1.5 molar equivalent, 25mM in DMA solution) were added sequentially, the DMA solution was supplemented, keeping the volume ratio of water to DMA in the system at 1:1, the system was reacted at 80℃for 1h.
Ethanol precipitation is carried out after the reaction is finished: adding 10% volume of 5M sodium chloride aqueous solution into the solution, continuously adding 3 times volume of cold ethanol, shaking uniformly, standing for 20 minutes at-20 ℃, centrifuging at 12000 speed for 5 minutes, pouring out supernatant, dissolving the rest precipitate with deionized water to obtain solution of On-DNA arylamine compound, quantifying by an enzyme-labeled instrument OD, and sending LCMS to confirm that the conversion rate of aryl iodide into arylamine is 98%.
EXAMPLE 2 Synthesis of On-DNA arylamine Compounds
The On-DNA aryl halide reacts with the aromatic amine under the catalysis of palladium to generate the On-DNA aromatic amine compound, and the specific reaction steps are as follows:
all solvents were sonicated for 30 minutes, to the On-DNA aryl halide solution (1 molar equivalent, 1mM in water), aqueous sodium hydroxide solution (300 molar equivalents, 1M in water), DMA solution of aromatic amine (80 molar equivalents, 1M in DMA), DMA solution of palladium catalyst (X molar equivalents, 12.5mM in DMA) and ligand (cBRIDP) (Y molar equivalents, 25mM in DMA) were added sequentially, keeping the DMA in the system at 18.75% of the solution volume and the system reacted at 80 ℃.
Ethanol precipitation is carried out after the reaction is finished: adding 10% volume of 5M sodium chloride aqueous solution into the solution, continuously adding 3 times volume of cold ethanol, shaking uniformly, standing for 20 minutes at-20 ℃, centrifuging at 12000 speed for 5 minutes, pouring out supernatant, dissolving the rest precipitate with deionized water to obtain solution of On-DNA arylamine compound, quantifying by an enzyme-labeled instrument OD, and sending LCMS to confirm the conversion rate of aryl halide into arylamine.
Table 1: example 2 table of yield data for reaction products under different reaction conditions.
From the above table it can be obtained: the On-DNA aryl halide is reacted with the aromatic amine BB1 to obtain higher yield, and the aromatic amine BB2 is reacted to obtain medium yield, 0.6 molar equivalent of [ (pi-all) PdCl is added] 2 And 1.5 molar equivalents of cBIDP were sufficient for the reaction system to reach equilibrium within 1h, see Table numbers 1 and 3, numbers 2 and 4.
EXAMPLE 3 Synthesis of On-DNA arylamine Compounds
The On-DNA aryl halide reacts with the aromatic amine under the catalysis of palladium to generate the On-DNA aromatic amine compound, and the specific reaction steps are as follows:
all solvents were sonicated for 30 min, to On-DNA aryl halide solution (1 molar equivalent, 1mM in water), aqueous sodium hydroxide solution (300 molar equivalents, 1M in water), DMA solution of aromatic amine (X molar equivalents, 1M in DMA), DMA solution of palladium catalyst (0.6 molar equivalents, 12.5mM in DMA) and ligand (cBRIDP) (1.5 molar equivalents, 25mM in DMA), DMA in the system accounting for Y% of the volume of the solution, and the system reacted at T ℃.
Ethanol precipitation is carried out after the reaction is finished: adding 10% volume of 5M sodium chloride aqueous solution into the solution, continuously adding 3 times volume of cold ethanol, shaking uniformly, standing for 20 minutes at-20 ℃, centrifuging at 12000 speed for 5 minutes, pouring out supernatant, dissolving the rest precipitate with deionized water to obtain solution of On-DNA arylamine compound, quantifying by an enzyme-labeled instrument OD, and sending LCMS to confirm the conversion rate of aryl halide into arylamine.
Table 2: example 3 yield data for reaction products under different conditions.
From the data in table 2 above, the optimal reaction conditions were found to be: the molar equivalent of the On-DNA aryl halide is 1, the molar equivalent of the aromatic amine added is 500, the DMA in the system accounts for 50 percent of the volume of the solution, 0.6 molar equivalent of palladium catalyst and 1.5 molar equivalent of ligand (cBIDP), the reaction temperature is 80 ℃, and the reaction time is 1h.
EXAMPLE 4 Synthesis of On-DNA arylamine Compounds
The On-DNA aryl iodide reacts with 71 aryl amines under the catalysis of palladium to generate the On-DNA aryl amine compound, and the specific reaction steps are as follows:
all solvents were sonicated for 30 min, to On-DNA aryl iodide solution (1 molar equivalent, 1mM in water), aqueous sodium hydroxide solution (300 molar equivalent, 1M in water), DMA solution of aromatic amine (500 molar equivalent, 1M in DMA), DMA solution of palladium catalyst (0.6 molar equivalent, 12.5mM in DMA) and ligand (cBRIDP) (1.5 molar equivalent, 25mM in DMA) were added sequentially, the DMA solution was supplemented, the volume ratio of water to DMA in the system was kept at 1:1, and the system was reacted at 80 ℃ for 1h.
Ethanol precipitation is carried out after the reaction is finished: adding 10% volume of 5M sodium chloride aqueous solution into the solution, continuously adding 3 times volume of cold ethanol, shaking uniformly, standing for 20 minutes at-20 ℃, centrifuging at 12000 speed for 5 minutes, pouring out supernatant, dissolving the rest precipitate with deionized water to obtain solution of On-DNA arylamine compound, quantifying by an enzyme-labeled instrument OD, and sending LCMS to confirm the conversion rate of aryl iodide into arylamine.
EXAMPLE 5 Synthesis of On-DNA arylamine Compounds
The On-DNA aryl bromide reacts with 59 kinds of aromatic amines under the catalysis of palladium to generate the On-DNA aromatic amine compound, and the specific reaction steps are as follows:
all solvents were sonicated for 30 min, to On-DNA aryl bromide solution (1 molar equivalent, 1mM aqueous solution), aqueous sodium hydroxide solution (300 molar equivalent, 1M aqueous solution), DMA solution of aromatic amine (500 molar equivalent, 1M DMA solution), DMA solution of palladium catalyst (0.6 molar equivalent, 12.5mM DMA solution) and ligand (cBRIDP) (1.5 molar equivalent, 25mM DMA solution) were added sequentially, and the DMA solution was supplemented, maintaining the volume ratio of water to DMA in the system at 1:1, the system was reacted at 80℃for 1h.
Ethanol precipitation is carried out after the reaction is finished: adding 10% volume of 5M sodium chloride aqueous solution into the solution, continuously adding 3 times volume of cold ethanol, shaking uniformly, standing for 20 minutes at-20 ℃, centrifuging at 12000 speed for 5 minutes, pouring out supernatant, dissolving the rest precipitate with deionized water to obtain solution of On-DNA arylamine compound, quantifying by an enzyme-labeled instrument OD, and sending LCMS to confirm the conversion rate of aryl halide into arylamine.
EXAMPLE 6 Synthesis of On-DNA arylamine Compounds
The On-DNA aryl chloride reacts with the aromatic amine under the catalysis of palladium to generate the On-DNA aromatic amine compound, and the specific reaction steps are as follows:
all solvents were sonicated for 30 min, to On-DNA aryl chloride solution (1 molar equivalent, 1mM in water), aqueous sodium hydroxide solution (300 molar equivalent, 1M in water), DMA solution of aromatic amine (500 molar equivalent, 1M in DMA), DMA solution of palladium catalyst (0.6 molar equivalent, 12.5mM in DMA) and ligand (cBRIDP) (1.5 molar equivalent, 25mM in DMA) were added sequentially, the DMA solution was supplemented, the volume ratio of water to DMA in the system was kept at 1:1, and the system was reacted at 80 ℃ for 1h.
Ethanol precipitation is carried out after the reaction is finished: adding 10% volume of 5M sodium chloride aqueous solution into the solution, continuously adding 3 times volume of cold ethanol, shaking uniformly, standing for 20 minutes at-20 ℃, centrifuging at 12000 speed for 5 minutes, pouring out supernatant, dissolving the rest precipitate with deionized water to obtain solution of On-DNA arylamine compound, quantifying by an enzyme-labeled instrument OD, and sending LCMS to confirm the conversion rate of aryl halide into arylamine.
The yield of 1c and BB1 was 88.7% and the yield of 1c and BB2 was 85.4%.
EXAMPLE 7 Synthesis of On-DNA arylamine Compounds
The On-DNA aryl chloride reacts with the aromatic amine under the catalysis of palladium to generate the On-DNA aromatic amine compound, and the specific reaction steps are as follows:
all solvents were sonicated for 30 min, to On-DNA aryl chloride solution (1 molar equivalent, 1mM in water), aqueous sodium hydroxide solution (300 molar equivalent, 1M in water), DMA solution of aromatic amine (500 molar equivalent, 1M in DMA solution), DMA solution of palladium catalyst (0.6 molar equivalent, 12.5mM in DMA solution) and ligand (cBRIDP) (1.5 molar equivalent, 25mM in DMA solution) were added in sequence, and the DMA solution was replenished, maintaining the volume ratio of water to N, N-dimethylacetamide in the system at 1:1, the system was reacted at 80℃for 1h.
Ethanol precipitation is carried out after the reaction is finished: adding 10% volume of 5M sodium chloride aqueous solution into the solution, continuously adding 3 times volume of cold ethanol, shaking uniformly, standing for 20 minutes at-20 ℃, centrifuging at 12000 speed for 5 minutes, pouring out supernatant, dissolving the rest precipitate with deionized water to obtain solution of On-DNA arylamine compound, quantifying by an enzyme-labeled instrument OD, and sending LCMS to confirm the conversion rate of aryl halide into arylamine.
The yield of the reaction of 1d and BB1 was 78.2%, and the reaction yield of 1d and BB2 was 84%.
EXAMPLE 8 Synthesis of On-DNA arylamine Compounds
The On-DNA aryl chloride reacts with the aromatic amine under the catalysis of palladium to generate the On-DNA aromatic amine compound, and the specific reaction steps are as follows:
all solvents were sonicated for 30 min, to On-DNA aryl chloride solution (1 molar equivalent, 1mM in water), aqueous sodium hydroxide solution (300 molar equivalent, 1M in water), DMA solution of aromatic amine (500 molar equivalent, 1M in DMA solution), DMA solution of palladium catalyst (0.6 molar equivalent, 12.5mM in DMA solution) and ligand (cBRIDP) (1.5 molar equivalent, 25mM in DMA solution) were added in sequence, and the DMA solution was replenished, keeping the volume ratio of water to DMA in the system at 1:1, the system was reacted at 80℃for 1h.
Ethanol precipitation is carried out after the reaction is finished: adding 10% volume of 5M sodium chloride aqueous solution into the solution, continuously adding 3 times volume of cold ethanol, shaking uniformly, standing for 20 minutes at-20 ℃, centrifuging at 12000 speed for 5 minutes, pouring out supernatant, dissolving the rest precipitate with deionized water to obtain solution of On-DNA arylamine compound, quantifying by an enzyme-labeled instrument OD, and sending LCMS to confirm the conversion rate of aryl halide into arylamine.
The reaction yield of 1e and BB1 was 90%, and the reaction yield of 1e and BB2 was 85%.
EXAMPLE 9 Synthesis of On-DNA arylamine Compounds
The On-DNA aryl chloride reacts with the aromatic amine under the catalysis of palladium to generate the On-DNA aromatic amine compound, and the specific reaction steps are as follows:
all solvents were sonicated for 30 min, to On-DNA aryl chloride solution (1 molar equivalent, 1mM in water), aqueous sodium hydroxide solution (1000 molar equivalent, 1M in water), DMA solution of aromatic amine (500 molar equivalent, 1M in DMA solution), DMA solution of palladium catalyst (0.6 molar equivalent, 12.5mM in DMA solution) and ligand (cBRIDP) (1.5 molar equivalent, 25mM in DMA solution) were added in sequence, and the DMA solution was supplemented, keeping the volume ratio of water to dimethylacetamide in the system at 1:1, the system was reacted at 80℃for 16h.
Ethanol precipitation is carried out after the reaction is finished: adding 10% volume of 5M sodium chloride aqueous solution into the solution, continuously adding 3 times volume of cold ethanol, shaking uniformly, standing for 20 minutes at-20 ℃, centrifuging at 12000 speed for 5 minutes, pouring out supernatant, dissolving the rest precipitate with deionized water to obtain solution of On-DNA arylamine compound, quantifying by an enzyme-labeled instrument OD, and sending LCMS to confirm the conversion rate of aryl halogen into arylamine.
The yield of the reaction of 1f and BB1 was 77%, and the reaction yield of 1f and BB2 was 56.4%.
In summary, the On-DNA aryl halogen compound can be converted into the On-DNA arylamine compound under the catalysis of the palladium reagent by controlling the conditions such as solvent, temperature, pH and the like during the reaction. The method has the advantages of few and easily available added reagent types, simple post-treatment, large-scale synthesis of On-DNA arylamine compounds, and suitability for constructing DNA coding compound libraries by using porous plates.
Claims (9)
1. A method for preparing arylamine compounds from On-DNA aryl halides, characterized by: the reaction is that an On-DNA aryl halogenated compound is taken as a substrate, and reacts with an amine compound under the action of alkali, a palladium catalyst and a ligand to generate an On-DNA aryl amine compound; the structural formula of the On-DNA aryl halogenated compound is DNA-Ar-X, and the structural formula of the amine compound is as follows:the structural formula of the On-DNA arylamine compound is->
Wherein the DNA in the formula comprises a single-stranded or double-stranded nucleotide chain polymerized from artificially modified and/or unmodified nucleotide monomers, the nucleotide chain being linked to Ar by one or more chemical bonds or groups; the length of the single-stranded or double-stranded nucleotide chain is 5-200 bp; -X of said On-DNA aryl halide is attached to the ring of Ar, X being chloro, bromo or iodo;
wherein R is 1 、R 2 Selected from hydrogen or a group having a molecular weight of 1000 or less directly attached to the amino nitrogen atom;
the reaction comprises the following steps: adding 5-1000 molar equivalents of alkali, 5-1000 molar equivalents of amine compound, 0.5-100 molar equivalents of palladium catalyst and 1-200 molar equivalents of ligand into an aryl halogenated compound solution with the molar concentration of 0.5-5mM, and reacting for 0.5-24 hours at the temperature of 20-100 ℃ to generate an On-DNA aromatic amine compound;
the palladium catalystThe chemical agent is [ (pi-all) PdCl] 2 Allyl palladium chloride dimer, the said ligand is cBRIDP;
the reaction solvent is a mixed solution of water and DMA;
the Ar is selected from the group consisting of:
2. the method according to claim 1, characterized in that: said R is 1 、R 2 Selected from hydrogen, alkyl, aryl, substituted aryl, aromatic heterocyclic group or substituted aromatic heterocyclic group; wherein the alkyl group is C 1 ～C 20 An alkyl group;
the aryl is selected from phenyl or naphthyl; the number of the substituent groups of the substituted aryl is one or more, and the substituent groups of the substituted aryl are one or more selected from halogen, cyano, carboxyl, alkoxy, halogen substituted alkoxy, alkyl, halogen substituted alkyl, aromatic heterocyclic group, alkyl substituted aromatic heterocyclic group and aminoacyl independently;
the aromatic heterocyclic group refers to a single cyclic group or a plurality of cyclic groups with aromaticity, wherein 5-10 of the cyclic groups contain C, O, S, N atoms; the number of the substituent groups of the substituted aromatic heterocyclic group is one or more, and the substituent groups of the substituted aromatic heterocyclic group are one or more selected from halogen, cyano, alkyl, halogen substituted alkyl, cycloalkyl, alkoxy, halogen substituted alkoxy, phenyl, halogenated phenyl, alkylphenyl, heterocyclic group, alkanoyl, carbonyl, sulfonyl and alkylsulfonyl which are independent of each other.
3. The process according to claim 1, wherein the base in the process is selected from sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, cesium carbonate, N-diisopropylethylamine or 1, 3-tetramethylguanidine.
4. The method of claim 1, wherein DMA is 10% to 60% of the solution volume.
5. The method of claim 1, wherein the reaction temperature is 40 ℃ to 90 ℃.
6. The method of claim 1, wherein the reaction time is 0.5 to 16 hours.
7. The method according to claim 1, wherein the molar equivalent of the amine compound is 300 to 1000.
8. The method according to any one of claims 1-7, wherein the method is used for batch multi-well plate operations.
9. The method according to any one of claims 1 to 7, wherein the method is used for the synthesis of a pool of DNA encoding compounds of a multiwell plate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010800122.4A CN114075257B (en) | 2020-08-11 | 2020-08-11 | Method for preparing arylamine compound from On-DNA aryl halide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010800122.4A CN114075257B (en) | 2020-08-11 | 2020-08-11 | Method for preparing arylamine compound from On-DNA aryl halide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114075257A CN114075257A (en) | 2022-02-22 |
| CN114075257B true CN114075257B (en) | 2023-08-22 |
Family
ID=80280061
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010800122.4A Active CN114075257B (en) | 2020-08-11 | 2020-08-11 | Method for preparing arylamine compound from On-DNA aryl halide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114075257B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113089104B (en) * | 2019-12-23 | 2023-03-28 | 成都先导药物开发股份有限公司 | Synthetic method of On-DNA diamine compound ring closure reaction |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004072088A2 (en) * | 2003-02-14 | 2004-08-26 | Takasago International Corporation | Phosphine compound, intermediate, palladium-complex, and use thereof |
| CN109071446A (en) * | 2016-01-29 | 2018-12-21 | 艾伯维德国有限责任两合公司 | The organic reaction carried out in aqueous solution in the presence of hydroxy alkyl (alkyl) cellulose or alkylcellulose |
| 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 |
| CN110240616A (en) * | 2018-03-07 | 2019-09-17 | 东莞市均成高新材料有限公司 | Three biaryl monophosphorus ligands, their preparation method and the purposes in catalyzed coupling reaction |
| CN110331447A (en) * | 2019-07-08 | 2019-10-15 | 上海药明康德新药开发有限公司 | The method of On-DNA aryl acetylene hydrocarbon compound is prepared in the building of DNA encoding compound library |
-
2020
- 2020-08-11 CN CN202010800122.4A patent/CN114075257B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004072088A2 (en) * | 2003-02-14 | 2004-08-26 | Takasago International Corporation | Phosphine compound, intermediate, palladium-complex, and use thereof |
| CN109071446A (en) * | 2016-01-29 | 2018-12-21 | 艾伯维德国有限责任两合公司 | The organic reaction carried out in aqueous solution in the presence of hydroxy alkyl (alkyl) cellulose or alkylcellulose |
| CN110240616A (en) * | 2018-03-07 | 2019-09-17 | 东莞市均成高新材料有限公司 | Three biaryl monophosphorus ligands, their preparation method and the purposes in catalyzed coupling reaction |
| 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 |
| CN110331447A (en) * | 2019-07-08 | 2019-10-15 | 上海药明康德新药开发有限公司 | The method of On-DNA aryl acetylene hydrocarbon compound is prepared in the building of DNA encoding compound library |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114075257A (en) | 2022-02-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112830950B (en) | Method for synthesizing On-DNA (deoxyribonucleic acid) dihydrothiazole/thiazole compound | |
| CN113089104B (en) | Synthetic method of On-DNA diamine compound ring closure reaction | |
| CN112837757A (en) | On-DNA Aldol reaction method in construction of DNA coding compound library | |
| CN113004361B (en) | Method for synthesizing On-DNA pyrazole compound | |
| CN114075257B (en) | Method for preparing arylamine compound from On-DNA aryl halide | |
| CN113072597B (en) | Method for synthesizing On-DNA carbonyl diaza-cycle compound | |
| CN112920245B (en) | Method for synthesizing On-DNA dihydropyrazole compound | |
| CN113563265B (en) | Method for synthesizing On-DNA N, N-monosubstituted indazolone compound | |
| CN112851732B (en) | Method for synthesizing On-DNA 2-carboxyl-3-amino arylthiophene compound | |
| CN113943979A (en) | Method for preparing On-DNA thioether compound from On-DNA aryl halide | |
| CN114957348B (en) | Method for synthesizing On-DNA (deoxyribonucleic acid) arylbenzotriazole compound and derivative thereof | |
| CN110627823B (en) | Method for catalyzing arylamine to generate deamination boric acid esterification or halogenation | |
| CN112921405B (en) | Method for synthesizing On-DNA pyrazolo [1,5-a ] pyrimidine compound | |
| CN114057817B (en) | Method for preparing arylboronic acid from On-DNA aryl halide | |
| CN112921406B (en) | Method for synthesizing On-DNA 2-aminopyrimidine compound | |
| CN112851718A (en) | Method for constructing On-DNA alpha-amino amide compound by aqueous phase Ugi multi-component reaction | |
| CN114411267B (en) | Method for constructing beta-aliphatic substituted ketone compound by On-DNA reaction | |
| CN112920246B (en) | Method for synthesizing On-DNA1,4-thiazepine compound | |
| CN112920244B (en) | Method for synthesizing On-DNA pyrimidine compound | |
| CN114478670B (en) | Method for synthesizing On-DNA beta substituted ketone compound | |
| CN116411356A (en) | Synthesis method of DNA coding compound ribofuranose derivative | |
| CN116732617A (en) | Method for constructing C-N bond by reacting On-DNA aryl vinyl compound with amine | |
| CN112779606A (en) | Method for On-DNA Mannich reaction | |
| CN116676674A (en) | Method for coupling C-C bonds in DNA coding compound library | |
| CN116411353A (en) | Synthesis method of DNA coding compound oxo-glycoside |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |