Synthesis method of on-dna benzodiazepine compound

Abstract

The application belongs to the technical field of DNA coded compound library, and particularly relates to a synthesis method of On-DNA benzodiazepine compounds, which comprises the following steps: taking On-DNA aldehyde compounds, diamine compounds and diketone compounds as substrates, and cyclizing to obtain On-DNA benzodiazepine compounds under mild and biocompatible reaction conditions, wherein the synthesis method is small in DNA damage, good in substrate universality, high in yield, low in cost, convenient in operation, suitable for the synthesis of On-DNA benzodiazepine compounds, rich in chemical reaction types of DEL library, and conducive to the drug development of benzodiazepine compounds.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of DNA-encoded compound library, and particularly relates to a synthesis method of on-DNA benzodiazepine compound. BACKGROUND

[0002] DNA-encoded library (DEL) technology is a new small molecule drug screening technology, which combines DNA technology with combinatorial chemistry, can efficiently construct a compound library with a capacity of hundreds of millions, and can simultaneously screen multiple or multiple conditions of the same target in the screening process. Compared with traditional high-throughput screening, DEL has great advantages in compound library capacity, library construction difficulty, screening time and cost.

[0003] In the on-DNA reaction for constructing DNA-encoded compound library, DNA must be stable in a certain aqueous phase, pH, temperature, metal ion concentration and inorganic salt concentration, which limits the types of reactions available in the construction of DNA-encoded compound library, resulting in limited molecular chemical structure diversity of the compound library. In addition, in addition to considering the compatibility with DNA chemistry, conversion rate is also a problem that those skilled in the art need to consider.

[0004] At present, the most commonly used bonding chemical reactions in the construction of DNA-encoded compound library are: amide bond formation reaction, reductive amination, aromatic nucleophilic substitution, Suzuki coupling reaction, Sonogashira coupling reaction, Heck coupling reaction, Buchwald coupling reaction, Ullmann coupling reaction and the like (reference https: / / delopen.org / reactions), and widening the types of chemical reactions for constructing DEL library is an important content for promoting the further development of DNA-encoded compound library technology.

[0005] Benzodiazepine compounds are a class of benzene seven-membered heterocyclic fused ring compounds, with special seven-membered ring structure and electronic state, most of which have strong physiological activity and medicinal value, and play a very important role in drug chemistry research. This kind of compounds can be used as inhibitors of various proteases, such as glucosidase inhibitors (New J. Chem., 2017, 41, 8993.), HIV protease inhibitors (J. Med. Chem., 2012, 55, 10130-10135..), HCV NS5B polymerase inhibitors (Bioorg. Med. Chem. Lett., 2009, 19, 2492-2496.) and the like. However, in the construction of DNA coded compound library, the synthesis method of on-DNA benzodiazepine compounds has not been reported so far. Therefore, we hope to develop a simple and fast method for synthesizing on-DNA benzodiazepine compounds. On the one hand, this research can enrich the chemical reaction types of DEL library construction, and on the other hand, this research can promote the understanding of the medicinal chemistry properties of benzodiazepine structure. SUMMARY

[0006] Therefore, the purpose of the present application is to provide a synthesis method of on-DNA benzodiazepine compounds, which can be applied to the construction of DNA coded compound library, enrich the chemical reaction types of DEL library construction, and promote the development of benzodiazepine compounds.

[0007] In order to achieve the above-mentioned purpose of the application, the technical scheme adopted by the present application is as follows:

[0008] A synthesis method of on-DNA benzodiazepine compounds, the synthesis method comprising:

[0009] providing an on-DNA aldehyde compound as shown in general formula (I), a diamine compound as shown in general formula (II) and a diketone compound as shown in general formula (III):

[0010]

[0011] reacting the on-DNA aldehyde compound, the diamine compound and the diketone compound in an aqueous solvent system to obtain an on-DNA benzodiazepine compound;

[0012] wherein, is DNA;

[0013] R 1 selected from -(R 5 ) m -(R 6 ) n -, m and n are not 0 at the same time, and R5 one selected from alkyl, alkoxy, cycloalkyl, R 6 one selected from aryl, substituted aryl, substituted heteroaryl, or heteroaryl, the substituents on the substituted aryl and the substituted heteroaryl being independently selected from one or more of alkyl, cycloalkyl, alkoxy, halogen, hydroxyl, trifluoromethyl, amino, ester, amide, nitro, cyano, phenyl;

[0014] R 2 one selected from hydrogen, alkyl, cycloalkyl, halogen, hydroxyl, alkoxy, trifluoromethyl, carboxyl, ester, amide, nitro, cyano, phenyl; or, the R 2 is fused to the benzene ring of the diamine compound to form a ring;

[0015] R 3 and R 4 are independently selected from alkyl or substituted alkyl, the substituents on the substituted alkyl being independently selected from one or more of carboxyl, halogen, hydroxyl, trifluoromethyl, amino, ester, amide, nitro, cyano, phenyl; or, the R 3 and the R 4 are connected to each other to form a ring.

[0016] In some embodiments, the R 6 is selected from phenyl, halophenyl, alkoxyphenyl, thienyl, furanyl, indolyl; and / or

[0017] the R 5 is selected from alkyl or alkoxy; and / or

[0018] n = 1, m = 1-3.

[0019] In some embodiments, the R 3 and the R 4 are connected to each other to form a ring.

[0020] In some embodiments, the solvent system consists essentially of an organic solvent and water, the organic solvent including at least one of acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, methanol, ethanol, t-butanol, isopropanol, tetrahydrofuran.

[0021] In some embodiments, in the step of reacting the On-DNA aldehyde compound, the diamine compound, and the diketone compound in an aqueous solvent system, the On-DNA aldehyde compound is reacted at a working concentration of 10-300 μΜ with less than 1000 molar equivalents of the diamine compound and 500-5000 molar equivalents of the diketone compound.

[0022] In some embodiments, the amount of the diamine compound is 50-500 molar equivalents.

[0023] In some embodiments, the amount of the diamine compound is 100-350 molar equivalents, and the amount of the diketone compound is 500-3500 molar equivalents.

[0024] In some embodiments, the step of reacting the On-DNA aldehyde compound, the diamine compound and the diketone compound in the aqueous solvent system comprises:

[0025] The diamine compound and the diketone compound are reacted in the solvent system to obtain a reaction system in which an intermediate product is dispersed, the temperature of the first reaction is greater than 25℃ and less than or equal to 100℃, and the time of the first reaction is 1-24 hours.

[0026] The On-DNA aldehyde compound is added to the reaction system to perform a second reaction, the temperature of the second reaction is greater than or equal to 20℃ and less than or equal to 100℃, and the time of the second reaction is 0.5-12 hours.

[0027] In some embodiments, the temperature of the first reaction is 30-90℃, and the time of the first reaction is 1-8 hours; and / or

[0028] The temperature of the second reaction is 20-40℃, and the time of the second reaction is 0.5-6 hours.

[0029] In the synthesis method of the on-DNA benzodiazepine compound provided by the present application, the On-DNA aldehyde compound, the diamine compound and the diketone compound are reacted in an aqueous solvent system to obtain the on-DNA benzodiazepine compound. At the same time, the reaction conditions are optimized to ensure a high conversion rate of the reaction.

[0030] The synthesis method of the present application has mild conditions, good biocompatibility, small damage to DNA, good substrate versatility, high yield, low cost, easy operation, is suitable for constructing a DNA coded compound library, enriches the type of DEL library, and is conducive to promoting the development of benzodiazepine compounds for drug development.

[0031] Therefore, the present application also provides a DNA coded compound library, which comprises the on-DNA benzodiazepine compound obtained by the above synthesis method. BRIEF DESCRIPTION OF DRAWINGS

[0032] Figure 1 The mass spectrum of the coded compound with serial number 6-1 in Example 1;

[0033] Figure 2 The mass spectra of the coded compounds numbered 6-3 in Example 1 are shown.

[0034] Figure 3 The mass spectra of the coded compounds numbered 6-4 in Example 1 are shown.

[0035] Figure 4 The mass spectra of the coded compounds numbered 6-5 in Example 1;

[0036] Figure 5 The mass spectra of the coded compounds numbered 6-6 in Example 1;

[0037] Figure 6 The mass spectra of the encoded compounds numbered 6-7 in Example 1 are shown.

[0038] Figure 7 The mass spectra of the encoded compounds numbered 6-8 in Example 1;

[0039] Figure 8 The mass spectra are for the coded compounds numbered 6-26 in Example 1. Detailed Implementation

[0040] In the description of this 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) nomenclature systems, and the specific compound groups involved are described and explained as follows:

[0041] "Alkyl" refers to a class of saturated chain hydrocarbon groups containing only carbon and hydrogen atoms, having straight-chain and / or branched carbon chains, including but not limited to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, isopentyl, hexyl, etc. In this invention, the number of carbon atoms in the alkyl group is preferably 1-6, and in some specific embodiments, the number of carbon atoms in the alkyl group is 1, 2, 3, 4, 5 or 6.

[0042] "Cycloalkyl" refers to a class of saturated hydrocarbon groups containing cyclic structures such as monocyclic, fused, spirocyclic, and bridged rings, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. In this 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.

[0043] "Alkoxy" refers to a class of alkyl groups that are directly bonded to an oxygen atom, including but not limited to methoxy, ethoxy, propoxy, butoxy, isobutoxy, tert-butoxy, etc. In this 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.

[0044] "Aryl" refers to any functional group or substituent derived from a simple aromatic ring, which can be monocyclic or polycyclic, including but not limited to phenyl, naphthyl, phenanthryl, anthraquinone, etc. The substituents in the substituted aryl group can be selected from C 1～6 alkyl, C 1～6 cycloalkyl, hydroxyl, halogen, alkoxy, trifluoromethyl, amino, aldehyde, ester, amide, cyano, etc.

[0045] "Phenyl" refers to a group of functional groups with a benzene ring, such as C6H5-, which can be substituted or unsubstituted benzene. The phenyl group in the present invention can be unsubstituted phenyl or substituted phenyl, and the substituents in the substituted phenyl group can be selected from C 1～6 alkyl, C 1～6 heteroalkyl, C 1～6 cycloalkyl, C 1～6 heterocycloalkyl, hydroxyl, halogen, alkoxy, trifluoromethyl, amino, aldehyde, ester, amide, cyano, etc.

[0046] "Heteroaryl" refers to a type of aryl group containing N, O, S, P, etc. The substituents in the substituted heteroaryl group can be selected from C 1～6 alkyl, C 1～6 cycloalkyl, hydroxyl, halogen, alkoxy, trifluoromethyl, amino, aldehyde, ester, amide, cyano, etc.

[0047] "Amide" refers to a group of groups containing amide bonds.

[0048] "Halogen" refers to the elements in Group VIIA of the periodic table, including chlorine (Cl), bromine (Br), iodine (I), etc.

[0049] "Amino" refers to NH2-.

[0050] "Hydroxyl" refers to a group consisting only of O and H, represented as -OH.

[0051] "Cyano" refers to a group consisting only of C and N, represented as -CN.

[0052] "Nitro" refers to a group consisting of N and O, represented as -NO2.

[0053] "Ester" refers to a group of groups containing ester bonds.

[0054] "Trifluoromethyl" refers to a methyl group with 3 hydrogen atoms replaced by fluorine atoms, represented as -CF3.

[0055] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present application more clearly understood, the present application will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are only intended to explain the present application, and not to limit the present application.

[0056] A synthesis method of an On-DNA benzodiazepine compound, comprising:

[0057] S01, providing an On-DNA aldehyde compound as shown in general formula (I), a diamine compound as shown in general formula (II) and a diketone compound as shown in general formula (III):

[0058]

[0059] S02, reacting the On-DNA aldehyde compound, the diamine compound and the diketone compound in an aqueous solvent system to obtain an On-DNA benzodiazepine compound;

[0060] wherein, is DNA;

[0061] R 1 is selected from -(R 5 ) m -(R 6 ) n -, m and n are not 0 at the same time, R 5 is selected from one of alkyl, alkoxy, cycloalkyl, R 6 is selected from aryl, substituted aryl, substituted heteroaryl or heteroaryl, and the substituents on the substituted aryl and the substituted heteroaryl are independently selected from one or more of alkyl, cycloalkyl, alkoxy, halogen, hydroxyl, trifluoromethyl, amino, ester, amide, nitro, cyano, phenyl;

[0062] R 2 is selected from one of hydrogen, alkyl, cycloalkyl, halogen, hydroxyl, alkoxy, trifluoromethyl, carboxyl, ester, amide, nitro, cyano, phenyl; or, R 2 is fused into a ring with the benzene ring of the diamine compound;

[0063] R 3 and R 4 are independently selected from alkyl or substituted alkyl, and the substituents on the substituted alkyl are independently selected from one or more of carboxyl, halogen, hydroxyl, trifluoromethyl, amino, ester, amide, nitro, cyano, phenyl; or, R 3 and R 4 are connected into a ring.

[0064] Specifically, in step S01, an On-DNA aldehyde compound, a diamine compound and a diketone compound are used as raw materials for synthesizing an On-DNA benzodiazepine compound.

[0065] In some embodiments, the R 6 is selected from one of phenyl, halophenyl, alkoxyphenyl, thienyl, furanyl, indolyl, the R 5 is selected from alkyl or alkoxy, n = 1, and m = 1-3.

[0066] In some embodiments, the R 3 and the R 4 are connected to each other to form a ring, which is:

[0067] In step S02, the On-DNA aldehyde compound, the diamine compound and the diketone compound are reacted in an aqueous solvent system to obtain an On-DNA benzodiazepine compound.

[0068] In some embodiments, the step of reacting the On-DNA aldehyde compound, the diamine compound and the diketone compound in an aqueous solvent system comprises:

[0069] S021, the diamine compound and the diketone compound are reacted in the solvent system to obtain a reaction system in which an intermediate product is dispersed, the temperature of the first reaction is greater than 25°C and less than or equal to 100°C, and the time of the first reaction is 1-24 hours;

[0070] S022, the On-DNA aldehyde compound is added to the reaction system to perform a second reaction, the temperature of the second reaction is greater than or equal to 20°C and less than or equal to 100°C, and the time of the second reaction is 0.5-12 hours.

[0071] In specific embodiments, the temperature of the first reaction is 30°C, 40°C, 50°C, 60°C, 70°C, 80°C, 90°C or 100°C, and the time is 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 10 hours, 12 hours, 15 hours, 18 hours, 20 hours, 23 hours or 24 hours. The temperature of the second reaction is 20°C, 25°C, 30°C, 35°C, 40°C, 50°C, 60°C, 70°C, 80°C or 90°C, and the time is 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 10 hours, 12 hours, 15 hours, 18 hours, 20 hours, 23 hours or 24 hours.

[0072] In further embodiments, the first reaction is carried out at a temperature of 30-90°C for 1-8 hours; and the second reaction is carried out at a temperature of 20-40°C for 0.5-6 hours. In this way, the yield of the synthesis is further improved.

[0073] The reaction between the on-DNA aldehyde compound, the diamine compound and the diketone compound is carried out in an aqueous solvent system. In some embodiments, the solvent system mainly consists of an organic solvent and water, and the organic solvent includes at least one of acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, dimethyl sulfoxide, methanol, ethanol, tert-butanol, isopropanol, and tetrahydrofuran. These organic solvents are good solvents, mild, and have good biocompatibility with substrates and little damage to DNA; and the blending with water can improve the yield of the synthesis reaction of the on-DNA benzodiazepine compound. In specific embodiments, the organic solvent is ethanol, acetonitrile or dimethyl sulfoxide, and the solvent system composed of these organic solvents and water has a higher yield.

[0074] In some embodiments, in the step of reacting the on-DNA aldehyde compound, the diamine compound and the diketone compound in an aqueous solvent system, the on-DNA aldehyde compound is reacted at a working concentration of 10-300 μM with less than 1000 molar equivalents of the diamine compound and 500-5000 molar equivalents of the diketone compound. In specific embodiments, the working concentration of the on-DNA aldehyde compound is 10 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, 100 μM, 110 μM, 120 μM, 130 μM, 140 μM, 150 μM, 160 μM, 170 μM, 180 μM, 190 μM, 200 μM, 210 μM, 220 μM, 230 μM, 240 μM, 250 μM, 260 μM, 270 μM, 280 μM, 290 μM or 300 μM. The molar equivalents of the diketone compound are 500 equivalents, 1000 equivalents, 1500 equivalents, 2000 equivalents, 2500 equivalents, 3000 equivalents, 3500 equivalents, 4000 equivalents, 4500 equivalents or 5000 equivalents. In further embodiments, the amount of the diamine compound is 50-500 molar equivalents, and in specific embodiments, the molar equivalents of the diamine compound are 50 equivalents, 100 equivalents, 150 equivalents, 200 equivalents, 250 equivalents, 300 equivalents, 350 equivalents, 400 equivalents, 450 equivalents or 500 equivalents. In further embodiments, the amount of the diamine compound is 100-350 molar equivalents, and the amount of the diketone compound is 500-3500 molar equivalents.

[0075] In summary, the synthesis method of the above On-DNA benzodiazepine compound provided by the embodiment of the present application is to react a diamine compound and a diketone compound in a solvent system containing water, and then react with an On-DNA aldehyde compound to obtain the On-DNA benzodiazepine compound. The synthesis method has mild conditions, good biocompatibility, small damage to DNA, good substrate universality, high yield, low cost, convenient operation, is suitable for constructing a DNA coded compound library, enriches the type of DEL library, and is conducive to promoting the drug development of the On-DNA benzodiazepine compound.

[0076] Based on the above technical solution, the present application further provides a DNA coded compound library, comprising: the On-DNA benzodiazepine compound synthesized by the above synthesis method.

[0077] The On-DNA benzodiazepine compound with an advantageous backbone structure generated by the synthesis method has good substrate universality, the three components are diverse, the conditions are mild, no catalyst is needed, the conversion rate is high, the post-treatment is simple, the operation is convenient, the environment is friendly, and the cost is low. The synthesized On-DNA benzodiazepine compound is suitable for constructing a DNA coded compound library, enriching the type of DEL library compound, and promoting the drug development of the benzodiazepine compound.

[0078] In order for the above implementation details and operations of the present application to be clearly understood by those skilled in the art, and the performance of the synthesis method of the On-DNA benzodiazepine compound provided by the embodiment of the present application is significantly embodied, the following examples are used to illustrate the implementation of the present application.

[0079] In the following examples, HATU: 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, EtOH: ethanol, ACN: acetonitrile, DMSO: dimethyl sulfoxide, HPLC: high performance liquid chromatograph, MS: mass spectrometer, LC-MS: liquid chromatograph-mass spectrometer, h: hour, min: minute, rt: room temperature.

[0080] Example 1

[0081] 1. Synthesis of On-DNA aldehyde compound (3)

[0082] The synthesis route is as follows:

[0083]

[0084] The specific synthesis steps are as follows:

[0085] The DNA-NH2 compound (1) was dissolved in borate buffer (250 mM, pH = 9.4) to prepare a solution of the compound (1) at a final concentration of 2.0 mM. The solution was reacted with an aldehyde carboxylic acid compound 2 using HATU as a condensing agent to obtain a corresponding On-DNA aldehyde compound (3). After the reaction was completed, the target product (On-DNA aldehyde compound (3)) was freeze-dried and directly used in the next reaction after ethanol precipitation treatment (specifically, 10% of a total reaction volume of a 5 M sodium chloride solution was added, 2.5 times the volume of anhydrous ethanol stored at -20°C, and the mixture was allowed to stand at -20°C for 1 h, and then centrifuged at 4°C at a speed of 13300 rpm for 15 min), HPLC purification, and MS detection.

[0086] 2. Synthesis of On-DNA benzodiazepine compounds (6)

[0087] The synthesis route is as follows:

[0088]

[0089] The specific synthesis steps are as follows:

[0090] 1) The On-DNA aldehyde compound (3) was dissolved in ultrapure water to prepare a solution at a concentration of 100 μM for use;

[0091] 2) In an EP tube, the diamine compound (4) (2 μL, 20 mM ethanol solution) and the diketone compound (5) (2 μL, 200 mM ethanol solution) were sequentially added, 14 μL of H2O was added, and the mixture was reacted at 60°C for 4 h;

[0092] 3) 2 μL of the On-DNA aldehyde compound (3) solution prepared in step 1) was added to the EP tube in step 2), and the mixture was reacted at 25°C for 1 h, followed by ethanol precipitation treatment, HPLC-MS detection, and calculation of the conversion rate of each small molecule.

[0093] Table 1 shows the chemical structures of representative On-DNA benzodiazepine compounds, their conversion rates, and molecular weight data.

[0094] Table 1

[0095]

[0096]

[0097]

[0098]

[0099]

[0100] Example 2

[0101] This example takes the compound (6-1) synthesized in Example 1 as a representative, and explores the influence of reaction temperature, reaction time, organic solvent system, and concentration relationship of reactants on the yield of the reaction according to the method steps of Example 1. Among them, the reaction temperature and reaction time refer to the reaction temperature and reaction time of the diamine compound (4) and the diketone compound (5) in Example 1.

[0102] The synthetic route is as follows:

[0103]

[0104] Table 2 is the test result, as shown in the result, the reaction temperature, reaction time, solvent system, and concentration relationship of reactants all have different degrees of influence on the yield of the reaction.

[0105] Among them, the reaction time of Example 1, 2, 3, 5 is 4h, and the solvent system of EtOH / H2O is used, and the concentration relationship between the reactants is also the same, the only difference is the different reaction temperature, and the reaction temperature of Example 1, Example 2 and Example 3 is 25℃, 40℃, 80℃ respectively, and the yield is also gradually increased, which is 0%, 49% and 87% respectively, and the yield of Example 5 is 90% at 60℃, which is greater than that of Example 3 (80℃), which shows that the yield of the reaction gradually increases with the increase of temperature, and when it reaches 60℃, the reaction can have a very high yield, and when it gradually increases to 80℃, it can still maintain a high yield.

[0106] The reaction temperature of Example 4, 5, 6 is 60℃, and the solvent system of EtOH / H2O is used, and the concentration relationship between the reactants is also the same, the only difference is the reaction time, when the reaction time reaches 4h, the yield can reach 90%, combined with the results of Example 4, 5, 6, it shows that when the reaction time reaches 4h, the reaction can have a high yield.

[0107] The difference between Example 5, 7, 8 is that the composition of the solvent system is different, the solvent system of Example 5 is EtOH / H2O, and the yield is 90%, the solvent system of Example 7 is ACN / H2O, and the yield is 62%, the solvent system of Example 8 is DMSO / H2O, and the yield is 76%, which shows that the use of different solvent systems has an impact on the yield, and thus the experiment can know that compared with the other two kinds of solvent system, the solvent system of EtOH / H2O has a better yield.

[0108] The difference between Examples 5, 9 and 10 is the concentration ratio between the reactants, the concentration ratio between the reactants in Example 9 is DNA-aldehyde (a): diamine (b): diketone (c) = 1:1000:2000, the yield is 0%; the concentration ratio between the reactants in Example 10 is DNA-aldehyde (a): diamine (b): diketone (c) = 1:200:1000, the yield is 65%; the concentration ratio between the reactants in Example 5 is DNA-aldehyde (a): diamine (b): diketone (c) = 1:200:2000, the yield is 90%, indicating that when the concentration of the reactants is DNA-aldehyde (a): diamine (b): diketone (c) = 1:200:2000, the yield is very high.

[0109] Table 2

[0110]

[0111] The above description is merely preferred embodiments of the present application, but not to limit the present application, any modification, equivalent replacement and improvement made within the spirit and principle of the present application should be included in the scope of protection of the present application.

Claims

1. A method for synthesizing an On-DNA benzodiazepine heterocyclic compound, wherein the general formula of the On-DNA benzodiazepine heterocyclic compound is as follows: ； The synthesis method includes: Provides on-DNA aldehydes as shown in general formula (I), diamines as shown in general formula (II), and diketones as shown in general formula (III): (I), (II), (III); The On-DNA aldehyde compound, the diamine compound, and the diketone compound are reacted in an aqueous solvent system to obtain an On-DNA benzodiazepine compound. in, DNA; R 1 Selected from -(R) 5 ) m -(R 6 ) n -, m and n are not both 0, the R 5 The R is selected from one of alkyl, alkoxy, and cycloalkyl groups. 6 The substituents are selected from aryl, substituted aryl, substituted heteroaryl, or heteroaryl, wherein the substituents on the substituted aryl and the substituted heteroaryl are independently selected from one or more of alkyl, cycloalkyl, alkoxy, halogen, hydroxyl, trifluoromethyl, amino, ester, amide, nitro, cyano, and phenyl. R 2 Selected from one of hydrogen, alkyl, cycloalkyl, halogen, hydroxyl, alkoxy, trifluoromethyl, carboxyl, ester, amide, nitro, cyano, and phenyl; or, said R 2 Fusing and forming a ring with the benzene ring of the diamine compound; R 3 and R 4 The groups are independently selected from alkyl or substituted alkyl groups, wherein the substituents on the substituted alkyl groups are independently selected from one or more of carboxyl, halogen, hydroxyl, trifluoromethyl, amino, ester, amide, nitro, cyano, and phenyl groups; or, the R 3 and the R 4 They are interconnected to form a ring.

2. The synthesis method according to claim 1, characterized in that, The R 6 Selected from one of phenyl, halophenyl, alkoxyphenyl, thiophene, furanyl, and indoleyl; and / or The R 5 Selected from alkyl or alkoxy; and / or n=1, m=1~3.

3. The synthesis method according to claim 1, characterized in that, The R 3 and the R 4 The interconnected loops are: , , , or .

4. The synthesis method according to claim 1, characterized in that, The solvent system mainly consists of organic solvents and water, and the organic solvents include at least one of acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, dimethyl sulfoxide, methanol, ethanol, tert-butanol, isopropanol, and tetrahydrofuran.

5. The synthesis method according to any one of claims 1 to 4, characterized in that, In the step of reacting the On-DNA aldehyde compound, the diamine compound, and the diketone compound in an aqueous solvent system, the On-DNA aldehyde compound at a working concentration of 10–300 μM is reacted with less than 1000 molar equivalents of the diamine compound and 500–5000 molar equivalents of the diketone compound.

6. The synthesis method according to claim 5, characterized in that, The amount of the diamine compound used is 50 to 500 molar equivalents.

7. The synthesis method according to claim 5, characterized in that, The amount of the diamine compound used is 100 to 350 molar equivalents, and the amount of the diketone compound used is 500 to 3500 molar equivalents.

8. The synthesis method according to any one of claims 1 to 4, characterized in that, The steps of reacting the On-DNA aldehyde compound, the diamine compound, and the diketone compound in an aqueous solvent system include: The diamine compound and the diketone compound are subjected to a first reaction in the solvent system to obtain a reaction system in which the intermediate product is dispersed. The temperature of the first reaction is greater than 25°C and less than or equal to 100°C, and the time of the first reaction is 1 to 24 hours. The On-DNA aldehyde compound is added to the reaction system to carry out a second reaction, the temperature of which is greater than or equal to 20°C and less than or equal to 100°C, and the reaction time is 0.5 to 12 hours.

9. The synthesis method according to claim 8, characterized in that, The first reaction is carried out at a temperature of 30–90°C for a time of 1–8 hours; and / or The temperature of the second reaction is 20–40°C, and the reaction time is 0.5–6 hours.

10. A DNA-encoded compound library, characterized in that, include: On-DNA benzodiazepine heterocyclic compounds obtained by the synthetic method according to any one of claims 1 to 9.

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