Method for preparing on-DNA aryl acetonitrile compound

Abstract

The invention belongs to the technical field of DNA coding compound libraries, and particularly relates to a method for preparing on-DNA aryl acetonitrile compounds. Specifically, the invention relates to the preparation method of the compound 5, the preparation method comprises the step of reacting a compound 3 with a compound 4 to prepare the compound 5, and the method has the advantages of simple operation, easily available raw materials and high yield. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the technical field of DNA-encoded compound libraries, and in particular relates to a method for preparing on-DNA aryl acetonitrile compounds. Background Art

[0002] DNA-encoded chemical libraries (DELs) have become an efficient method for lead discovery in early drug discovery research. Generally, the construction of small molecules in DEL production relies not only on the availability of various building blocks, but also on established DNA-compatible chemical methods. In order to expand the scope of the chemical space available for drug discovery and identify more bioactive molecules, it is very necessary to develop new DNA-compatible reactions. However, the presence of the DNA backbone imposes several intrinsic limitations, such as DNA integrity (protection from strong oxidants, strong reducing agents, strong acids, etc.) and DNA solubility issues (highly diluted aqueous media). One of the most important tasks in the field of DEL libraries is the development of chemical reactions on DNA, referred to as On-DNA chemical reactions. Because DNA must be stable in a certain aqueous phase, pH, temperature, metal ion concentration and inorganic salt concentration, On-DNA chemical reactions with little DNA damage, good recovery rate and wide substrate adaptability are what are needed for large-scale application in the synthesis of DNA-encoded compound libraries.

[0003] Nitrile-containing compounds have attracted significant attention due to their wide applications in pharmaceutical science. In particular, several important aromatic acetonitrile drugs, such as verapamil, anastrozole, and gallopamil, have been widely developed as prescription drugs. On the other hand, the nitrile group can also serve as a valuable synthon in organic synthetic chemistry and be converted into other useful functional groups including aldehydes, carboxylic acids, and amines, which can be further used in DEL synthesis. However, to the best of our knowledge, the introduction of nitrile groups into organic molecules with oligodeoxynucleotide barcodes remains challenging.

[0004] The present disclosure develops a new DNA-compatible reaction for the cyanomethylation of (hetero)aryl halides via a tandem process involving palladium-mediated Suzuki-Miyaura coupling and base-promoted isoxazole fragmentation. The one-pot protocol exhibits a broad substrate application range, employs readily available starting materials, and does not cause significant DNA damage. Furthermore, the resulting (hetero)aryl acetonitriles can be converted to the corresponding carboxylic acids, which can be further used for the synthesis of DNA-encoded chemical libraries. Summary of the invention

[0005] The present disclosure relates to a method for preparing a compound 5, comprising the following steps B: reacting a compound 3 with a compound 4 to prepare a compound 5,

[0006]

[0007] wherein L is selected from a bond or a linking group;

[0008] R a Selected from hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-10 Cycloalkyl or 3-10 membered heterocycloalkyl;

[0009] X is selected from Cl, Br or I;

[0010] R 1 Selected from halogen, NO2, OH, NH2, COOH, N(C 1-6 Alkyl)2, NH(C 1-6 Alkyl), COOC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Alkyl O-, C 1-6 Alkyl S-, C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 5-10 Cycloalkenyl, C 6-10 Aryl or 5-10 membered heteroaryl, the N(C 1-6 Alkyl)2, NH(C 1-6 Alkyl), COOC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Alkyl O-, C 1-6 Alkyl S-, C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 5-10 Cycloalkenyl, C 6-10 The aryl or 5-10 membered heteroaryl is optionally substituted by one or more groups selected from the group consisting of halogen, NO2, OH or NH2;

[0011] n is selected from 0, 1, 2, 3 or 4;

[0012] Ring A is selected from C 6-10 Aryl, 5-10 membered heteroaryl, C 3-10 Cycloalkyl, C 5-10 Cycloalkenyl or 5-12 membered heterocyclic group.

[0013] In some embodiments, the linking group of L is such that the shortest distance between the N atom to which it is attached and ring A is no more than 12 atoms, 11 atoms, 10 atoms, 9 atoms, 8 atoms, 7 atoms, 6 atoms, 5 atoms, 4 atoms, 3 atoms, 2 atoms, 1 atom, or 0 atoms.

[0014] In some embodiments, the linking group of L is such that the shortest distance between the N atom to which it is attached and ring A is 0-12 atoms, or 0-8 atoms, or 0-6 atoms, or 0-4 atoms, or 0-2 atoms, or 0-1 atom.

[0015] In some embodiments, the linker group of L is such that the atom connected to the N atom connected thereto and the ring A with the shortest distance therebetween is selected from C, N, O or S.

[0016] In some embodiments, the linking group of L is such that, among the connecting atoms with the shortest distance between the N atom connected thereto and the ring A, the substituents of the same or different atoms can be connected to each other to form a ring structure.

[0017] In some embodiments, the linking group of L allows different atoms in the linking atom with the shortest distance between the N atom connected thereto and ring A to be connected to each other to form a ring structure.

[0018] In some embodiments, the ring structure is selected from C 6-10 aryl, 5-10 membered heteroaryl, 5-12 membered cycloalkyl, 5-12 membered cycloalkenyl or 5-12 membered heterocyclyl.

[0019] In some embodiments, the linking group of L is such that among the connecting atoms with the shortest distance between the N atom connected thereto and ring A, the different atoms do not form a ring structure.

[0020] In some embodiments, L is selected from a bond, -CO-, -CO-L a -、-SO-、-SO-L a -, -S(O)2- or -S(O)2-L a -, where L a Select from linking groups.

[0021] In some embodiments, the L a The connecting group is such that the shortest distance between the -CO-, -SO- or -S(O)2- group connected thereto and ring A is no more than 12 atoms, 11 atoms, 10 atoms, 9 atoms, 8 atoms, 7 atoms, 6 atoms, 5 atoms, 4 atoms, 3 atoms, 2 atoms, 1 atom or 0 atoms.

[0022] In some embodiments, the L aThe connecting group is such that the shortest distance between the -CO-, -SO- or -S(O)2- group connected thereto and ring A is 0-12 atoms, or 0-8 atoms, or 0-6 atoms, or 0-4 atoms, or 0-2 atoms, or 0-1 atom.

[0023] In some embodiments, the L a The connecting group is such that among the connecting atoms with the shortest distance between the -CO-, -SO- or -S(O)2- group connected thereto and ring A, the atom is selected from C, N, O or S.

[0024] In some embodiments, the L a The connecting group is such that among the connecting atoms with the shortest distance between the -CO-, -SO- or -S(O)2- group connected thereto and ring A, the substituents of the same or different atoms may be connected to each other to form a ring structure.

[0025] In some embodiments, the L a The connecting group allows the connecting atoms with the shortest distance between the -CO-, -SO- or -S(O)2- group connected thereto and the ring A to be connected, and the different atoms can be connected to each other to form a ring structure.

[0026] In some embodiments, the ring structure is selected from C 6-10 aryl, 5-10 membered heteroaryl, 5-12 membered cycloalkyl, 5-12 membered cycloalkenyl or 5-12 membered heterocyclyl.

[0027] In some embodiments, the L a The connecting group is such that among the connecting atoms with the shortest distance between the -CO-, -SO- or -S(O)2- group connected thereto and ring A, the different atoms do not form a ring structure.

[0028] In some embodiments, L is selected from a bond, -CO-, -CO-L a -、-SO-、-SO-L a -, -S(O)2- or -S(O)2-L a -, where L a Selected from -C 1-6 Alkyl-, -C 2-6 Alkenyl-, -C 2-6 Alkynyl-, -C 3-10 Cycloalkyl- or -3-10 membered heterocycloalkyl-.

[0029] In some embodiments, L is selected from a bond, -CO- or -CO-L a -, where L a Selected from -C1-6 Alkyl-, -C 2-6 Alkenyl-, -C 2-6 Alkynyl-, -C 3-10 Cycloalkyl- or -3-10 membered heterocycloalkyl-.

[0030] In some embodiments, L a Selected from -C 1-4 Alkyl-, -C 2-4 Alkenyl-, -C 2-4 Alkynyl-, -C 3-6 Cycloalkyl- or -3-6 membered heterocycloalkyl-.

[0031] In some embodiments, L a Selected from -C 1-3 Alkyl-, -C 2-3 Alkenyl-, -C 2-3 Alkynyl- or -C 3-6 Cycloalkyl-.

[0032] In some embodiments, L a Selected from -C 1-2 Alkyl-, -C 2-3 Alkenyl or -C 3-4 Cycloalkyl-.

[0033] In some embodiments, L a Selected from -CH2-, -CH=CH- or -cyclopropyl-.

[0034] In some embodiments, L is selected from a bond, -CO-, -COCH2-, -COCH=CH-, or -CO-cyclopropyl-.

[0035] In some embodiments, R a Selected from hydrogen, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl.

[0036] In some embodiments, R a Selected from hydrogen, C 1-3 Alkyl, C 2-3 Alkenyl, C 2-3 Alkynyl or C 3-6 Cycloalkyl.

[0037] In some embodiments, R a Selected from hydrogen, C 1-2 Alkyl, C 2-3 Alkenyl or C 3-4 Cycloalkyl.

[0038] In some embodiments, Ra Selected from hydrogen.

[0039] In some embodiments, X is selected from Br or I. In some embodiments, X is selected from Br.

[0040] In some embodiments, R 1 Selected from halogen, NO2, OH, NH2, COOH, N(C 1-6 Alkyl)2, NH(C 1-6 Alkyl), COOC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Alkyl O-, C 3-10 Cycloalkyl or 3-10 membered heterocycloalkyl, the N(C 1-6 Alkyl)2, NH(C 1-6 Alkyl), COOC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Alkyl O-, C 3-10 The cycloalkyl or 3-10 membered heterocycloalkyl is optionally substituted with one or more groups selected from halogen, NO2, OH or NH2.

[0041] In some embodiments, R 1 Selected from halogen, NO2, OH, NH2, COOH, N(C 1-4 Alkyl)2, NH(C 1-4 Alkyl), COOC 1-4 Alkyl, C 1-4 Alkyl, C 1-4 Alkyl O-, C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl, the N(C 1-4 Alkyl)2, NH(C 1-4 Alkyl), COOC 1-4 Alkyl, C 1-4 Alkyl, C 1-4 Alkyl O-, C 3-6 The cycloalkyl or 3-6 membered heterocycloalkyl is optionally substituted with one or more groups selected from halogen, NO2, OH or NH2.

[0042] In some embodiments, R 1 Selected from F, Cl, Br, I, NO2, OH, NH2, COOH, N(C 1-3 Alkyl)2, NH(C 1-3 Alkyl), COOC 1-3 Alkyl, C 1-3 Alkyl, C 1-3 Alkyl O-, C 5-6 Cycloalkyl or 5-6 membered heterocycloalkyl, the N(C 1-3Alkyl)2, NH(C 1-3 Alkyl), COOC 1-3 Alkyl, C 1-3 Alkyl, C 1-3 Alkyl O-, C 5-6 The cycloalkyl or 5-6 membered heterocycloalkyl is optionally substituted with one or more halogens.

[0043] In some embodiments, R 1 Selected from F, Cl, Br, I, NO2, OH, NH2, COOH, C 1-3 Alkyl, C 1-3 alkylO- or 6-membered heterocycloalkyl, the C 1-3 Alkyl or C 1-3 The alkyl O- is optionally substituted with one or more F, Cl or Br.

[0044] In some embodiments, R 1 Selected from F, Cl, NO2, COOH, C 1-3 Alkyl, C 1-3 alkylO- or 6-membered heterocycloalkyl, the C 1-3 Alkyl or C 1-3 Alkyl O- is optionally substituted with one or more F or Cl.

[0045] In some embodiments, R 1 Selected from F, Cl, NO2, COOH, CH3-, CH3O- or a 6-membered heterocycloalkyl group containing N and / or O, wherein the CH3- or CH3O- is optionally substituted by one or more F.

[0046] In some embodiments, R 1 Selected from F, Cl, NO2, COOH, CH3-, CF3-, CH3O-, CF3O- or morpholinyl.

[0047] In some embodiments, n is selected from 0, 1, 2, or 3. In some embodiments, n is selected from 0, 1, or 2.

[0048] In some embodiments, n is selected from 0 or 1.

[0049] In some embodiments, Ring A is selected from C 6-10 aryl, 5-10 membered heteroaryl, or 5-12 membered heterocyclyl.

[0050] In some embodiments, Ring A is selected from C 6-10 aryl, 5-10 membered heteroaryl or 5-10 membered heterocyclyl.

[0051] In some embodiments, Ring A is selected from C 6-10 aryl, 5-10 membered heteroaryl or 5-10 membered heterocyclyl.

[0052] In some embodiments, Ring A is selected from C 6-10 aryl, 5-10 membered heteroaryl or 5-10 membered dicycloheterocyclic group.

[0053] In some embodiments, Ring A is selected from phenyl, naphthyl, 5-membered heteroaryl, 6-membered heteroaryl, 9-membered heteroaryl, 10-membered heteroaryl, benzo-5-membered heterocyclyl, or 6-membered heteroaryl-6-membered heterocyclyl.

[0054] In some embodiments, ring A is selected from phenyl, naphthyl, furanyl, thienyl, pyrazolyl, pyridinyl, pyrimidinyl,

[0055] In some embodiments, the structural fragment DNA is selected from

[0056]

[0057] In some embodiments, the above preparation method further comprises the preparation step A of compound 3: compound 1 reacts with compound 2 to prepare compound 3.

[0058]

[0059] R is selected from halogen, COOH, -L a -COOH, -S(O)OH, -L a -S(O)OH, -S(O)2OH or -L a -S(O)2OH;

[0060] Ring A, X, R 1 , L a , n and L are as defined in the present disclosure.

[0061] In some embodiments, compound 1 is selected from

[0062] In some embodiments, compound 3 is selected from

[0063] In some embodiments, compound 5 is selected from

[0064] In some embodiments, compound 1 is selected from

[0065] In some embodiments, R is selected from halogen, COOH or -L a In some embodiments, R is selected from Cl, Br, I, COOH or -L a COOH.

[0066] In some embodiments, R is selected from Cl, COOH, -CH2COOH, -CH=CHCOOH, or -cyclopropyl-COOH.

[0067] In some embodiments, Compound 2 is selected from the following structures:

[0068]

[0069] In some embodiments, compound 3 is selected from the following structures:

[0070]

[0071]

[0072] In some embodiments, compound 5 is selected from the following structures:

[0073]

[0074]

[0075] The present disclosure relates to a method for preparing a compound 5, comprising the following steps: Step A: Compound 1 reacts with compound 2 to prepare compound 3;

[0076]

[0077] Step B: Compound 3 reacts with compound 4 to prepare compound 5.

[0078]

[0079] Among them, ring A, X, R 1 , R a , R, n and L are as defined in the present disclosure.

[0080] In some embodiments, step B of the above preparation method is carried out in the presence of a base. In some embodiments, the base is selected from sodium carbonate, potassium phosphate, cesium hydroxide, potassium carbonate, or potassium fluoride. In some embodiments, the base is selected from sodium carbonate, potassium phosphate, cesium hydroxide or potassium carbonate. In some embodiments, the base is selected from sodium carbonate or cesium hydroxide. In some embodiments, the molar ratio of the base to compound 3 is selected from 100 to 1000: 1; or selected from 100 to 800: 1; or selected from 100 to 600: 1; or selected from 200 to 500: 1; or selected from 200 to 400: 1. In some embodiments, the molar ratio of the base to compound 3 is selected from 200: 1, 300: 1 or 400: 1.

[0081] In some embodiments, step B of the preparation method is carried out in the presence of a catalyst. In some embodiments, the catalyst is selected from a palladium-containing catalyst. In some embodiments, the catalyst is selected from RuPhos-Pd-G2, RuPhos-Pd-G4 or sSPhos-Pd-G2. In some embodiments, the molar ratio of the catalyst to compound 3 is selected from 1 to 20:1; or selected from 1 to 15:1; or selected from 2 to 10:1. In some embodiments, the molar ratio of the catalyst to compound 3 is selected from 2:1, 6:1 or 10:1.

[0082] In some embodiments, the preparation method step B is carried out in the presence of a solvent. In some embodiments, the solvent is selected from one or more of DMF, DMSO, DMAC, isopropanol or water. In some embodiments, the solvent is selected from DMF aqueous solution, DMSO aqueous solution, DMAC aqueous solution or isopropanol aqueous solution.

[0083] In some embodiments, in step B, compound 3 is dissolved in water to be added to the reaction system. In some embodiments, compound 4 is dissolved in a 50% (v:v) DMF aqueous solution or a 50% (v:v) isopropanol aqueous solution to be added to the reaction system. In some embodiments, the catalyst is dissolved in a DMF solution to be added to the reaction system.

[0084] In some embodiments, step B of the preparation method comprises vortexing and centrifuging the reaction system.

[0085] In some embodiments, step B of the above preparation method is carried out at 80-100°C; or at 90-95°C; or at 95°C; in some embodiments, in the above preparation method, step B is reacted for 0.5-10 hours; or 1-8 hours; or 2-5 hours; or 3-4 hours; or 3 hours.

[0086] In some embodiments, in the above preparation method, step B comprises adding sodium diethyldithiocarbamate trihydrate. Optionally, sodium diethyldithiocarbamate trihydrate is added and vortexed. Further optionally, the system is reacted at 70 to 90°C; or at 80°C. Optionally, the reaction time is selected from 20 minutes to 60 minutes; or the reaction time is selected from 20 minutes to 40 minutes; or the reaction time is 30 minutes.

[0087] In some embodiments, in the above preparation method, the molar ratio of compound 4 to compound 3 in step B is selected from 50 to 500: 1; or 50 to 400: 1; or 100 to 300: 1. In some embodiments, in the above preparation process, the molar ratio of compound 4 to compound 3 is selected from 100: 1, 200: 1 or 300: 1.

[0088] In some embodiments, the above preparation method, step B comprises: adding compound 3 (1.0 eq), base (300.0 eq), compound 4 (50% N, N-dimethylformamide aqueous solution, 200.0 eq) and catalyst (6.0 eq) to the reaction tube in sequence. Optionally, the mixture is vortexed for 5 seconds, centrifuged at 1000 r / min for 1 minute at 4°C, and reacted at 95°C for 3 hours. Optionally, sodium diethyldithiocarbamate trihydrate (300.0 eq) is added to the reaction system, the mixture is vortexed for 5 seconds, centrifuged at 1000 r / min for 1 minute at 4°C, and reacted at 80°C for 0.5 hours. Optionally, after the reaction is completed, 5M sodium chloride solution and anhydrous ethanol are added for post-treatment. Further, vortexing and centrifugation are optionally included. Optionally, after the reaction is completed, 5M sodium chloride solution and anhydrous ethanol refrigerated at minus 20°C are added, the mixture is vortexed for 5 seconds, centrifuged at 1000r / min for 1 minute at 4°C, placed at minus 80°C for 0.5 hours, and centrifuged at 4°C for 30 minutes at 4000r / min, and the supernatant is removed. Optionally, after the reaction is completed, 5M sodium chloride solution and anhydrous ethanol refrigerated at minus 20°C are added, the mixture is vortexed for 5 seconds, centrifuged at 1000r / min for 1 minute at 4°C, placed at minus 80°C for 0.5 hours, centrifuged at 4°C for 30 minutes at 4000r / min, the supernatant is removed, and an 80% ethanol aqueous solution refrigerated at minus 20°C is added, the mixture is vortexed for 5 seconds, centrifuged at 1000r / min for 1 minute at 4°C, placed at minus 80°C for 0.5 hours, centrifuged at 4°C for 30 minutes at 4000r / min, and the supernatant is removed.

[0089] Optionally, the compound 5 obtained in step B is lyophilized and then prepared into a 1.0 mM aqueous solution.

[0090] In some embodiments, the above preparation method, step A is carried out in the presence of boric acid. Optionally, the boric acid is present in the form of a boric acid buffer. In some embodiments, the concentration of boric acid is 250 mM.

[0091] In some embodiments, in the above preparation method, in step A, compound 1 is mixed with boric acid buffer and then added to the reaction system. Optionally, the concentration of compound 1 in the boric acid buffer is 1.0 mM.

[0092] In some embodiments, step A of the above preparation method is carried out in the presence of a condensing agent or a base; in some embodiments, step A of the above preparation method is carried out in the presence of 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride or sodium bicarbonate.

[0093] In some embodiments, the above preparation method, step A is carried out in the presence of a solvent. In some embodiments, the solvent is selected from one or more of DMF, DMSO or DMAC. In some embodiments, the solvent is selected from DMAC.

[0094] In some embodiments, step A of the preparation method comprises vortexing and centrifuging the reaction system.

[0095] In some embodiments, the step A of the above preparation method is carried out at room temperature. Optionally, the reaction time is selected from 10 to 15 hours; or 10 to 12 hours; or 12 hours.

[0096] In some embodiments, in the above preparation method, step A comprises adding piperidine. Optionally, after adding piperidine, vortexing and centrifugation are performed. Further, optionally, the reaction is carried out at room temperature for 20 minutes to 60 minutes; or for 20 minutes to 40 minutes; or for 30 minutes.

[0097] In some embodiments, in the above preparation method, the molar ratio of compound 1 to compound 2 in step A is selected from 1:20-200; or 1:30-150; or 1:40-100; or 1:40-50; or 1:80-120; or 1:80-100.

[0098] In some embodiments, in the above preparation method, the molar ratio of compound 1 to compound 2 in step A is selected from 1:40 or 1:100.

[0099] In another aspect, the present disclosure provides Compound 3 or a pharmaceutically acceptable salt thereof,

[0100]

[0101] Among them, ring A, X, R 1 , R a , n and L are as defined in the present disclosure.

[0102] In another aspect, the present disclosure provides the following compound or a pharmaceutically acceptable salt thereof:

[0103]

[0104]

[0105] In another aspect, the present disclosure provides Compound 5 or a pharmaceutically acceptable salt thereof,

[0106]

[0107] Among them, ring A, R 1 , R a, n and L are as defined in the present disclosure.

[0108] In another aspect, the present disclosure provides the following compound or a pharmaceutically acceptable salt thereof:

[0109]

[0110]

[0111] In yet another aspect, the present disclosure provides use of the compound described in the present disclosure in an On-DNA chemical reaction.

[0112] Technical Effects

[0113] The present invention discloses a novel preparation method of on-DNA cycloacetonitrile compounds, especially aryl acetonitrile compounds. DNA can remain stable in a certain aqueous phase, pH, temperature, metal ion concentration and inorganic salt concentration, has little damage to DNA, has a good recovery rate and wide substrate adaptability, mild reaction conditions, cheap and easy-to-obtain raw materials, is suitable for large-scale production, and the preparation method has simple reaction operation, few impurities and high yield.

[0114] definition

[0115] The term "alkyl" refers to a group of the formula C n H 2n+1 The alkyl group may be straight chain or branched. For example, the term "C 1-6 The term "alkyl" refers to an alkyl group containing 1 to 6 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc.). Similarly, the alkyl portion (i.e., alkyl) of alkoxy, alkylamino, dialkylamino, alkylsulfonyl, and alkylthio has the same definition as above. The term "alkylene" refers to a divalent group formed by removing one hydrogen at any position of an alkyl group. The same may be Straight or branched chain, with two positions attached to the rest of the molecule. For example, the term "-C0-6 alkylene-" refers to an alkylene group containing 0 to 6 carbons, non-limiting examples of which include, but are not limited to, -CH2-, -CH2CH2-, -CH(CH3)-, -CH2CH2CH2-, -CH(CH3)CH2-, -C(CH3)2-, -CH2CH2CH2CH2-, etc. C0 represents a bond. Alkyl and alkylene groups may be unsubstituted or substituted with one or more substituents.

[0116] In this article, C m-n , means that the moiety has an integer number of carbon atoms in a given range. For example, "C 1-6" means that the group may have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms or 6 carbon atoms. For example, C 1-3 It means that the group may have 1 carbon atom, 2 carbon atoms, or 3 carbon atoms.

[0117] The term "alkenyl" refers to a linear or branched unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms having at least one double bond. Non-limiting examples of alkenyl include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, 1-butenyl, isobutenyl, 1,3-butadienyl, etc.

[0118] The term "alkynyl" refers to a straight or branched unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms having at least one triple bond. Non-limiting examples of alkynyl include, but are not limited to, ethynyl (-C≡CH), 1-propynyl (-C≡C-CH3), 2-propynyl (-CH2-C≡CH), 1,3-butadiynyl (-C≡CC≡CH), etc.

[0119] The term "alkoxy" refers to an -O-alkyl group.

[0120] The term "halo" or "halogen" refers to fluorine, chlorine, bromine and iodine.

[0121] The term "cycloalkyl" refers to a fully saturated carbocyclic ring that can exist as a monocyclic, bridged or spirocyclic ring. Unless otherwise indicated, the carbocyclic ring is typically a 3 to 10-membered ring (e.g., a 3-8-membered, 4-7-membered, 4-6-membered or 5-6-membered ring). Non-limiting examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl (bicyclo[2.2.1]heptyl), bicyclo[2.2.2]octyl, adamantyl, etc.

[0122] The term "heterocyclic radical" refers to a non-aromatic ring that is fully saturated or partially unsaturated (but not fully unsaturated heteroaromatic) and can exist as a monocyclic, bridged or spirocyclic ring. Unless otherwise indicated, the heterocyclic ring is generally 3 to 7 rings (e.g., 3, 4, 5, 6 or 7 rings) containing 1 to 3 heteroatoms (preferably 1 or 2 heteroatoms) independently selected from sulfur, oxygen and / or nitrogen. Non-limiting examples of heterocyclic radicals include but are not limited to oxirane, tetrahydrofuranyl, dihydrofuranyl, pyrrolidinyl, N-methylpyrrolidinyl, dihydropyrrolyl, piperidinyl, piperazinyl, pyrazolidinyl, 4H-pyranyl, morpholinyl, thiomorpholinyl, tetrahydrothienyl etc.

[0123] The term "heterocycloalkyl" refers to a cyclic group that is fully saturated and can exist as a monocyclic, bridged or spirocyclic ring. Unless otherwise indicated, the heterocycle is typically a 3 to 10-membered ring (e.g., 3-8, 4-7, 4-6 or 5-6-membered ring) containing 1 to 3 heteroatoms (preferably 1 or 2 heteroatoms) independently selected from sulfur, oxygen and / or nitrogen. Examples of 3-membered heterocycloalkyl include, but are not limited to, oxirane, thioethane, and cyclonitroethane. Non-limiting examples of 4-membered heterocycloalkyl include, but are not limited to, azetidinyl, oxadiazolyl, and thiadinyl. Examples of 5-membered heterocycloalkyl include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, isoxazolidinyl, oxazolidinyl, isothiazolidinyl, thiazolidinyl, imidazolidinyl, and tetrahydropyrazolyl. Examples of 6-membered heterocycloalkyl include, but are not limited to, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, piperazinyl, 1,4-thioxanyl, 1,4-dioxanyl, thiomorpholinyl, 1,3-dithianyl, and 1,4-dithianyl. Examples of 7-membered heterocycloalkyl include, but are not limited to, azepanyl, oxepanyl, and thiepanyl. Preferably, the monocyclic heterocycloalkyl has 5 or 6 ring atoms.

[0124] The term "aryl" refers to an all-carbon monocyclic or fused polycyclic aromatic ring group having a conjugated π electron system. For example, an aryl group may have 6-20 carbon atoms, 6-14 carbon atoms, or 6-12 carbon atoms. Non-limiting examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, and 1,2,3,4-tetrahydronaphthalene, etc.

[0125] The term "heteroaryl" refers to a monocyclic or fused polycyclic system containing at least one ring atom selected from N, O, S, the remaining ring atoms being C, and having at least one aromatic ring. Preferred heteroaryls have a single 5- to 8-membered ring, or multiple fused rings containing 6 to 14, especially 6 to 10, ring atoms. Non-limiting examples of heteroaryls include, but are not limited to, pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, quinolyl, isoquinolyl, tetrazolyl, triazolyl, triazinyl, benzofuranyl, benzothienyl, indolyl, isoindolyl, etc.

[0126] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may be substituted or not substituted, and unless otherwise specified, the type and number of the substituents may be arbitrary on the basis of chemical achievable.

[0127] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0128] When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may be optionally substituted with up to two Rs, and each occurrence of R is an independent choice. In addition, combinations of substituents and / or variants thereof are permitted only if such combinations result in stable compounds.

[0129] When a substituent is vacant, it means that the substituent does not exist. For example, when X in AX is vacant, it means that the structure is actually A. When the listed substituent does not specify which atom it is connected to the substituted group through, the substituent can be bonded through any atom of it. For example, pyridyl as a substituent can be connected to the substituted group through any carbon atom on the pyridine ring.

[0130] Unless otherwise specified, the number of atoms in the ring is usually defined as the number of ring members, for example, "5-7 membered ring" refers to a "ring" with 5-7 atoms arranged around it. The following abbreviations are used in this application:

[0131] eq means molar equivalent. DMF means N,N-dimethylformamide. DMSO means dimethyl sulfoxide. DMAC means N,N-dimethylacetamide. DETAILED DESCRIPTION

[0132]

[0133] Embodiment 1:

[0134]

[0135] Step 1: Preparation of intermediate 3a:

[0136] Compound 1 (i.e., HP-P11-AOP-NH2, 100.0 nmol, 100.0 μL, 1.0 mM, borate buffer (250 mM, pH = 9.5), 1.0 eq), 4-bromobenzoic acid 2a (10000 nmol, 50.0 μL, 200 mM N, N-dimethylacetamide solution, 100.0 eq) and 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (10000 nmol, 50.0 μL, 200 mM aqueous solution, 100.0 eq) were added to the PCR reaction tube in sequence. The mixture was vortexed for 5 seconds, centrifuged at 1000 r / min for 1 minute at 4°C, and reacted at 25°C for 12 hours.

[0137] Piperidine (20.0 μL) was added to the reaction system, the mixture was vortexed for 5 seconds, centrifuged at 1000 r / min for 1 minute at 4°C, and reacted at 25°C for 0.5 hours. After the reaction was completed, 5M sodium chloride solution (22.0 μL) and anhydrous ethanol (550.0 μL) refrigerated at -20°C were added, the mixture was vortexed for 5 seconds, centrifuged at 1000 r / min for 1 minute at 4°C, placed at -80°C for 0.5 hours, centrifuged at 4000 r / min for 30 minutes at 4°C, and the supernatant was removed. The obtained product was separated and purified by reverse phase high performance liquid chromatography, lyophilized to obtain intermediate 3a, and then configured into a 1.0 mM aqueous solution for standby use. LC-MS: m / z 12218.8.

[0138] Step 2: Preparation of compound 5a

[0139] Compound 3a (10.0 nmol, 10.0 μL, 1.0 mM aqueous solution, 1.0 eq), sodium carbonate (3000 nmol, 3.0 μL, 1.0 M aqueous solution, 300.0 eq), intermediate 4 (isoxazole-4-boronic acid, 2000.0 nmol, 10.0 μL, 200 mM 50% N, N-dimethylformamide aqueous solution, 200.0 eq) and sSPhos-Pd-G2 (60.0 nmol, 6.0 μL, 10 mM N, N-dimethylformamide solution, 6.0 eq) were added to the PCR reaction tube in sequence. The mixture was vortexed for 5 seconds, centrifuged at 1000 r / min for 1 minute at 4°C, and reacted at 95°C for 3 hours. Sodium diethyldithiocarbamate trihydrate (3000.0 nmol, 10.0 μL, 300 mM aqueous solution, 300.0 eq) was added to the reaction system, and the mixture was vortexed for 5 seconds, centrifuged at 1000 r / min at 4° C. for 1 minute, and reacted at 80° C. for 0.5 hours.

[0140] After the reaction was completed, 5 M sodium chloride solution (3.9 μL) and anhydrous ethanol (97.5 μL) refrigerated at -20°C were added, the mixture was vortexed for 5 seconds, centrifuged at 1000 r / min for 1 minute at 4°C, placed at -80°C for 0.5 hours, centrifuged at 4000 r / min for 30 minutes at 4°C, and the supernatant was removed. After the reaction was completed, 5M sodium chloride solution (22.0 μL) and anhydrous ethanol (550.0 μL) refrigerated at -20°C were added, the mixture was vortexed for 5 seconds, centrifuged at 1000 r / min for 1 minute at 4°C, placed at -80°C for 0.5 hours, centrifuged at 4°C for 30 minutes at 4000 r / min, the supernatant was removed, and 80% ethanol aqueous solution (50.0 μL) refrigerated at -20°C was added, the mixture was vortexed for 5 seconds, centrifuged at 1000 r / min for 1 minute at 4°C, placed at -80°C for 0.5 hours, centrifuged at 4°C for 30 minutes at 4000 r / min, and the supernatant was removed. The obtained product (5a) was lyophilized and then configured into a 1.0 mM aqueous solution. The mass spectrum was measured, and the yield was calculated by TIC, and the yield of on-DNA compound 5a of Example 1 was 84%.

[0141] Examples 2 to 18: Referring to step 2 of Example 1, compound 3a is reacted with compound 4, and the reagents are adjusted accordingly as shown in the following table (the feed ratio refers to the molar ratio of the corresponding reagent (base, solvent or catalyst) or compound (4) to compound 3a), as shown in Table 1 below.

[0142] Table 1

[0143]

[0144]

[0145] Examples 19 to 60: Referring to steps 1 and 2 of Example 1, compound 2a in step 1 was replaced with the following compounds 2b to 2aq, and the products corresponding to step 2 were finally prepared. As shown in Table 2 below.

[0146] Table 2

[0147]

[0148]

[0149]

[0150]

[0151]

[0152] Example 61:

[0153]

[0154] Step 1: Preparation of intermediate 3ar

[0155] Compound 1 (i.e., HP-P11-AOP-NH2, 100.0 nmol, 100.0 μL, 1.0 mM boric acid buffer (250 mM, pH = 9.5), 1.0 eq), 2,4-dichloro-6-methoxypyrimidine 2ar (4000.0 nmol, 20.0 μL, 200 mM N, N-dimethylacetamide solution, 40.0 eq) and sodium bicarbonate (4000.0 nmol, 20.0 μL, 200 mM aqueous solution, 40.0 eq) were added to the PCR reaction tube in sequence. The mixture was vortexed for 5 seconds, centrifuged at 1000 r / min for 1 minute at 4°C, and reacted at 80°C for 4 hours.

[0156] After the reaction is completed, take a sample for mass spectrometry. After the reaction is completed, add 5M sodium chloride solution (14.0μL) and anhydrous ethanol (350.0μL) refrigerated at -20°C, vortex the mixture for 5 seconds, centrifuge at 1000r / min for 1 minute at 4°C, place at -80°C for 0.5 hours, centrifuge at 4000r / min for 30 minutes at 4°C, and remove the supernatant. The obtained product is separated and purified by reverse phase high performance liquid chromatography, and after freeze drying, it is configured into a 1.0mM aqueous solution for standby use. LC-MS: m / z 12178.1.

[0157] Step 2: Preparation of compound 5ar

[0158] Compound 3ar (10.0 nmol, 10.0 μL, 1.0 mM aqueous solution, 1.0 eq), sodium carbonate (3000.0 nmol, 3.0 μL, 1.0 M aqueous solution, 300.0 eq), intermediate 4 (isoxazole-4-boronic acid, 2000.0 nmol, 10.0 μL, 200 mM 50% N, N-dimethylformamide aqueous solution, 200.0 eq) and sSPhos-Pd-G2 (60.0 nmol, 6.0 μL, 10 mM N, N-dimethylformamide solution, 6.0 eq) were added to the PCR reaction tube in sequence. The mixture was vortexed for 5 seconds, centrifuged at 1000 r / min for 1 minute at 4°C, and reacted at 95°C for 3 hours.

[0159] Sodium diethyldithiocarbamate trihydrate (3000.0 nmol, 10.0 μL, 300 mM aqueous solution, 300.0 eq) was added to the reaction system, and the mixture was vortexed for 5 seconds, centrifuged at 1000 r / min for 1 minute at 4°C, and reacted at 80°C for 0.5 hours. After the reaction was completed, 5M sodium chloride solution (3.9 μL) and anhydrous ethanol (97.5 μL) refrigerated at -20°C were added, and the mixture was vortexed for 5 seconds, centrifuged at 1000 r / min for 1 minute at 4°C, placed at -80°C for 0.5 hours, centrifuged at 4°C at 4000 r / min for 30 minutes, and the supernatant was removed. After the reaction was completed, 5M sodium chloride solution (22.0 μL) and anhydrous ethanol (550.0 μL) refrigerated at -20°C were added, the mixture was vortexed for 5 seconds, centrifuged at 1000 r / min for 1 minute at 4°C, placed at -80°C for 0.5 hours, centrifuged at 4°C for 30 minutes at 4000 r / min, the supernatant was removed, and 80% ethanol aqueous solution (50.0 μL) refrigerated at -20°C was added, the mixture was vortexed for 5 seconds, centrifuged at 1000 r / min for 1 minute at 4°C, placed at -80°C for 0.5 hours, centrifuged at 4°C for 30 minutes at 4000 r / min, and the supernatant was removed. The obtained product (5ar) was freeze-dried and configured into a 1.0 mM aqueous solution. The mass spectrum was measured, and the yield was calculated by TIC, and the yield of on-DNA compound 5ar was 39%.

[0160] Examples 62-64: Referring to steps 1 and 2 of Example 61, the compound 2ar in step 1 was replaced with the following compounds 2as to 2au, respectively, as shown in Table 3 below.

[0161] Table 3

[0162]

[0163]

[0164] Table 4 Mass spectral data of compounds

[0165] Compound LC-MS: m / z Compound LC-MS: m / z Compound LC-MS: m / z Compound LC-MS: m / z 5a 12178.5 5m 12219.1 5y 12218.7 5ak 12179.0 5b 12179.1 5n 12223.0 5z 12217.8 5al 12179.0 5c 12208.6 5o 12229.2 5aa 12235.0 5am 12192.3 5d 12208.7 5p 12222.8 5ab 12218.9 5an 12208.8 5e 12263.2 5q 12168.8 5ac 12218.7 5ao 12196.9 5f 12197.3 5r 12184.5 5ad 12230.1 5ap 12212.8 5g 12247.4 5s 12168.7 5ae 12230.0 5aq 12219.6 5h 12223.6 5t 12183.0 5af 12230.6 5ar 12183.1 5i 12223.9 5u 12179.7 5ag 12218.6 5as 12208.8 5j 12263.9 5v 12179.5 5ah 12218.6 5at 12205.0 5k 12192.9 5w 12180.5 5ai 12219.9 5au 12191.5 5l 12211.1 5x 12194.6 5aj 12218.5 .

Claims

1. A method for preparing compound 5, comprising the following steps B: reacting compound 3 with compound 4 to prepare compound 5, in, L is selected from a bond or a linking group; R a Selected from hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-10 Cycloalkyl or 3-10 membered heterocycloalkyl; X is selected from Cl, Br or I; R 1 Selected from halogen, NO2, OH, NH2, COOH, N(C 1-6 Alkyl)2, NH(C 1-6 Alkyl), COOC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Alkyl O-, C 1-6 Alkyl S-, C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 5-10 Cycloalkenyl, C 6-10 Aryl or 5-10 membered heteroaryl, the N(C 1-6 Alkyl)2, NH(C 1-6 Alkyl), COOC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Alkyl O-, C 1-6 Alkyl S-, C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 5-10 Cycloalkenyl, C 6-10 The aryl or 5-10 membered heteroaryl is optionally substituted by one or more groups selected from the group consisting of halogen, NO2, OH or NH2; n is selected from 0, 1, 2, 3 or 4; Ring A is selected from C 6-10 Aryl, 5-10 membered heteroaryl, C 3-10 Cycloalkyl, C 5-10 Cycloalkenyl or 5-12 membered heterocyclic group.

2. The preparation method according to claim 1, wherein the linking group of L is such that the shortest distance between the N atom connected thereto and the ring A is no more than 12 atoms, 11 atoms, 10 atoms, 9 atoms, 8 atoms, 7 atoms, 6 atoms, 5 atoms, 4 atoms, 3 atoms, 2 atoms, 1 atom or 0 atoms; Optionally, the linking group of L is such that among the connecting atoms with the shortest distance between the N atom connected thereto and the ring A, the substituents of the same or different atoms can be connected to each other to form a ring structure; Optionally, L is selected from a bond, -CO-, -CO-L a -、-SO-、-SO-L a -, -S(O)2- or -S(O)2-L a -, where L a is selected from a linking group; Optionally, the L a The connecting group is such that the shortest distance between the -CO-, -SO- or -S(O)2- group connected thereto and the ring A is no more than 12 atoms, 11 atoms, 10 atoms, 9 atoms, 8 atoms, 7 atoms, 6 atoms, 5 atoms, 4 atoms, 3 atoms, 2 atoms, 1 atom or 0 atoms; Optionally, the L a The connecting group is such that among the connecting atoms with the shortest distance between the -CO-, -SO- or -S(O)2- group connected thereto and the ring A, the substituents of the same or different atoms can be connected to each other to form a ring structure; Optionally, L is selected from a bond, -CO-, -CO-L a -、-SO-、-SO-L a -, -S(O)2- or -S(O)2-L a -, where L a Selected from -C 1-6 Alkyl-, -C 2-6 Alkenyl-, -C 2-6 Alkynyl-, -C 3-10 Cycloalkyl- or -3-10 membered heterocycloalkyl-; Optionally, L is selected from a bond, -CO- or -CO-L a -, where L a Selected from -C 1-6 Alkyl-, -C 2-6 Alkenyl-, -C 2-6 Alkynyl-, -C 3-10 Cycloalkyl- or -3-10 membered heterocycloalkyl-; Optionally, L a Selected from -C 1-4 Alkyl-, -C 2-4 Alkenyl-, -C 2-4 Alkynyl-, -C 3-6 Cycloalkyl- or -3-6 membered heterocycloalkyl-; Optionally, L a Selected from -C 1-3 Alkyl-, -C 2-3 Alkenyl-, -C 2-3 Alkynyl- or -C 3-6 Cycloalkyl-; Optionally, L a Selected from -C 1-2 Alkyl-, -C 2-3 Alkenyl or -C 3-4 Cycloalkyl-; Optionally, L a is selected from -CH2-, -CH=CH- or -cyclopropyl-; Alternatively, L is selected from a bond, -CO-, -COCH2-, -COCH=CH- or -CO-cyclopropyl-.

3. The preparation method according to claim 1 or 2, wherein R a Selected from hydrogen, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl; Optionally, R a Selected from hydrogen, C 1-3 Alkyl, C 2-3 Alkenyl, C 2-3 Alkynyl or C 3-6 Cycloalkyl; Optionally, R a Selected from hydrogen, C 1-2 Alkyl, C 2-3 Alkenyl or C 3-4 Cycloalkyl; Optionally, R a Selected from hydrogen.

4. The preparation method according to any one of claims 1 to 3, wherein R 1 Selected from halogen, NO2, OH, NH2, COOH, N(C 1-6 Alkyl)2, NH(C 1-6 Alkyl), COOC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Alkyl O-, C 3-10 Cycloalkyl or 3-10 membered heterocycloalkyl, the N(C 1-6 Alkyl)2, NH(C 1-6 Alkyl), COOC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Alkyl O-, C 3-10 Cycloalkyl or 3-10 membered heterocycloalkyl is optionally substituted by one or more groups selected from: halogen, NO2, OH or NH2; Optionally, R 1 Selected from halogen, NO2, OH, NH2, COOH, N(C 1-4 Alkyl)2, NH(C 1-4 Alkyl), COOC 1-4 Alkyl, C 1-4 Alkyl, C 1-4 Alkyl O-, C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl, the N(C 1-4 Alkyl)2, NH(C 1-4 Alkyl), COOC 1-4 Alkyl, C 1-4 Alkyl, C 1-4 Alkyl O-, C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl is optionally substituted by one or more groups selected from: halogen, NO2, OH or NH2; Optionally, R 1 Selected from F, Cl, Br, I, NO2, OH, NH2, COOH, N(C 1-3 Alkyl)2, NH(C 1-3 Alkyl), COOC 1-3 Alkyl, C 1-3 Alkyl, C 1-3 Alkyl O-, C 5-6 Cycloalkyl or 5-6 membered heterocycloalkyl, the N(C 1-3 Alkyl)2, NH(C 1-3 Alkyl), COOC 1-3 Alkyl, C 1-3 Alkyl, C 1-3 Alkyl O-, C 5-6 Cycloalkyl or 5-6 membered heterocycloalkyl is optionally substituted with one or more halogens; Optionally, R 1 Selected from F, Cl, Br, I, NO2, OH, NH2, COOH, C 1-3 Alkyl, C 1-3 alkylO- or 6-membered heterocycloalkyl, the C 1-3 Alkyl or C 1-3 Alkyl O- is optionally substituted with one or more F, Cl or Br; Optionally, R 1 Selected from F, Cl, NO2, COOH, C 1-3 Alkyl, C 1-3 alkylO- or 6-membered heterocycloalkyl, the C 1-3 Alkyl or C 1-3 Alkyl O- is optionally substituted with one or more F or Cl; Optionally, R 1 Selected from F, Cl, NO2, COOH, CH3-, CH3O- or a 6-membered heterocycloalkyl group containing N and / or O, wherein the CH3- or CH3O- is optionally substituted with one or more F; Optionally, R 1 Selected from F, Cl, NO2, COOH, CH3-, CF3-, CH3O-, CF3O- or morpholinyl.

5. The preparation method according to any one of claims 1 to 4, wherein ring A is selected from C 6-10 Aryl, 5-10 membered heteroaryl, or 5-12 membered heterocyclyl; Optionally, Ring A is selected from C 6-10 Aryl, 5-10 membered heteroaryl or 5-10 membered heterocyclyl; Optionally, Ring A is selected from C 6-10 Aryl, 5-10 membered heteroaryl or 5-10 membered heterocyclyl; Optionally, Ring A is selected from C 6-10 Aryl, 5-10 membered heteroaryl or 5-10 membered dicyclic heterocyclic group; Optionally, ring A is selected from phenyl, naphthyl, 5-membered heteroaryl, 6-membered heteroaryl, 9-membered heteroaryl, 10-membered heteroaryl, benzo 5-membered heterocyclyl or 6-membered heteroaryl and 6-membered heterocyclyl; Optionally, ring A is selected from phenyl, naphthyl, furanyl, thienyl, pyrazolyl, pyridinyl, pyrimidinyl, 6. The preparation method according to any one of claims 1 to 5, further comprising the step A of preparing compound 3: reacting compound 1 with compound 2 to prepare compound 3, R is selected from halogen, COOH, -L a -COOH, -S(O)OH, -L a -S(O)OH, -S(O)2OH or -L a -S(O)2OH; Optionally, R is selected from halogen, COOH or -L a COOH; Optionally, R is selected from Cl, Br, I, COOH or -L a COOH; Optionally, R is selected from Cl, COOH, -CH2COOH, -CH=CHCOOH or -cyclopropyl-COOH.

7. The preparation method according to any one of claims 1 to 6, wherein step B is carried out in the presence of a base; Optionally, the base is selected from sodium carbonate, potassium phosphate, cesium hydroxide, potassium carbonate, or potassium fluoride; Optionally, the molar ratio of the base to compound 3 is selected from 100 to 1000:1; or selected from 100 to 800:1; or selected from 100 to 600:1; or Selected from 200 to 500:1; or selected from 200 to 400:1; Optionally, the molar ratio of the base to compound 3 is selected from 200:1, 300:1 or 400:1; Optionally, step B is carried out in the presence of a catalyst; Optionally, the catalyst is selected from a catalyst containing palladium; Optionally, the catalyst is selected from RuPhos-Pd-G2, RuPhos-Pd-G4 or sSPhos-Pd-G2; Optionally, the molar ratio of the catalyst to compound 3 is selected from 1 to 20:1; or selected from 1 to 15:1; or selected from 2 to 10:1; Optionally, the molar ratio of the catalyst to compound 3 is selected from 2:1, 6:1 or 10:1; Optionally, step B is carried out in the presence of a solvent; Optionally, the solvent is selected from one or more of DMF, DMSO, DMAC, isopropanol or water; Optionally, the solvent is selected from DMF aqueous solution, DMSO aqueous solution, DMAC aqueous solution or isopropanol aqueous solution; Optionally, step B of the preparation method is carried out at 80-100° C.; or at 90-95° C.; or at 95° C.; step B is reacted for 0.5-10 hours; or 1-8 hours; or 2-5 hours; or 3-4 hours; or 3 hours; Optionally, step B comprises adding sodium diethyldithiocarbamate trihydrate.

8. The preparation method according to any one of claims 1 to 7, wherein in step B, the molar ratio of compound 4 to compound 3 is selected from 50 to 500:1; or 50~400:1; or 100~300:1; Optionally, the molar ratio of compound 4 to compound 3 is selected from 100:1, 200:1 or 300:

1.

9. The preparation method according to any one of claims 6 to 8, wherein step A is carried out in the presence of boric acid; Optionally, the boric acid is present in the form of a boric acid buffer; Optionally, step A is carried out in the presence of a condensing agent or a base; Optionally, step A thereof is carried out in the presence of 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride or sodium bicarbonate.

10. Compound 3, Compound 5 or a pharmaceutically acceptable salt thereof: in, Ring A, R 1 , R a , n and L are defined as in any one of claims 1-5.

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