Tetrahydroisoquinoline compounds as potassium channel regulators and their preparation and use

Abstract

The present invention relates to tetrahydroisoquinoline compounds as potassium channel modulators and their preparation and use. Specifically, the compounds of the present invention have a structure shown in formula A, wherein the definitions of the groups and substituents are as described in the specification. The present invention also discloses a preparation method of the compounds and their use as potassium channel modulators.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to tetrahydroisoquinoline compounds as potassium channel regulators and preparation and application thereof. Background Art

[0002] Kv7 potassium channel is a type of voltage-dependent potassium ion channel with the characteristics of low threshold activation, slow activation and non-inactivation. Kv7 potassium channel has five family members (Kv7.1-Kv7.5), all of which have similar topological structures, that is, a functional channel composed of four subunits, each of which contains six transmembrane segments (S1-S6). Among them, S4 is the voltage sensing region, which plays an important role in sensing membrane potential changes and controlling conformational changes; SS-S6 is the main component of the channel pore region, which is the main combination and action area of ​​potassium channel openers. KV7.1 potassium channel is a non-neuronal pathway distributed in peripheral tissues and expressed in the heart to mediate myocardial Iks. Its mutation can lead to Long QT syndrome. Kv7.2-Kv7.5 potassium channels are the basis of neuronal M currents, widely distributed in the nervous system, and have multiple physiological activities. Mutations in the Kv7.2 and Kv7.3 potassium channel genes can lead to a variety of different epilepsy phenotypes, such as benign familial neonatal convulsions (BFNC), which fully demonstrates the role of M currents in regulating neuronal excitability. Kv7.4 potassium channels are highly expressed in the outer hair cells of the cochlea and the auditory nucleus of the brainstem, and their mutations may cause hereditary deafness. Kv7.5 potassium channels are highly expressed in skeletal muscle and brain, and their mutations may cause retinopathy. Many diseases such as epilepsy, anxiety, and deafness have a common feature of high membrane excitability. Kv7 potassium channels, as the molecular basis of M currents, can open by sensing changes in membrane potential, upregulating inhibitory potassium currents, and thus controlling membrane excitability, making Kv7 potassium channels important in pain and mental illnesses represented by high neural excitability.

[0003] Retigabine is a drug for the treatment of epilepsy and has been approved for marketing in the UK, Germany and Denmark. Studies have shown that the effects of retigabine are related to voltage-gated potassium channels (KCNQs), and its main mechanism of action is to regulate M-type potassium currents through KCNQ2 / 3 channels.

[0004] Retigabine (RTG) is the first Kv7 potassium channel opener for the adjuvant treatment of partial-onset epilepsy in adults, which was launched in 2011. In addition to its anti-epileptic effect, RTG can also be used to treat anxiety, neuralgia, neurodegenerative diseases, etc. RTG can effectively reduce or prevent epileptic seizures in a variety of epilepsy models. RTG shows effective anti-epileptic effects on tonic seizures caused by the maximum electric shock (MES) model and clonic seizures induced by PTZ. In addition, RTG can also prevent epileptic seizures caused by N-methyl-D-aspartate (NMDA), penicillin, picrotoxin, kainic acid (KA), etc. The kindling model is suitable for the screening of a variety of anti-epileptic drugs, and the effect of RTG on this model is stronger than other models. Due to the wide range of effects of RTG on all Kv7 potassium channel members and other channels, it has poor selectivity, which makes it have potential adverse effects. A large number of literature reports show that the incidence of adverse events related to the central nervous system of RTG is high, which can cause dizziness, fatigue, aphasia, speech disorders, balance disorders, and other adverse reactions. Other adverse reactions include kidney and urinary system diseases such as kidney stones and urinary retention, cardiac arrest, transient non-sustained ventricular tachycardia and other heart-related diseases. It can also cause retinal discoloration, blue / purple pigmentation of the skin and nails, etc. Summary of the invention

[0005] The object of the present invention is to provide a compound represented by formula A, a preparation method thereof and use thereof as a potassium channel regulator.

[0006] In a first aspect of the present invention, there is provided a compound represented by formula A or a pharmaceutically acceptable salt thereof,

[0007]

[0008] in,

[0009] R1 is selected from the following substituted or unsubstituted groups: C 6-10 Aryl, 4-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S, wherein the substitution refers to substitution by one or more substituents selected from the group consisting of halogen, nitro, cyano, C 1-6 Alkyl, C 3-6 Cycloalkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyloxy, halogenated C 1-6 Alkyl, halogenated C 3-6 Cycloalkyl, halogenated C 1-6 Alkoxy, halogenated C 3-6 Cycloalkyloxy, -NR8R9, ethynyl;

[0010] R2 and R3 are independently selected from the following groups: hydrogen, deuterium, halogen, C1-6 Alkyl, halogenated C 1-6 alkyl;

[0011] or R2 and R3 and their respective connected C form C 3-10 Cycloalkyl, the cycloalkyl is optionally substituted by 1-3 substituents selected from the group consisting of hydrogen, halogen, C 1-6 Alkyl, halogenated C 1-6 alkyl;

[0012] R4 and R5 are independently selected from the following groups: hydrogen, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy;

[0013] R6 is selected from the following substituted or unsubstituted groups: C 1-6 Alkyl, C 3-6 Cycloalkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyloxy, the substitution refers to substitution by one or more substituents selected from the group consisting of halogen, nitro, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl;

[0014] n is selected from the following group: 1, 2, 3;

[0015] W is C-R7 or N;

[0016] R7 is selected from the group consisting of hydrogen, deuterium, halogen, cyano, C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy;

[0017] R8 and R9 are independently selected from the following groups: hydrogen, C 1-6 Alkyl, halogenated C 1-6 alkyl;

[0018] Or R8 and R9 and the N to which they are attached form a 3-10 membered azacycloalkyl group.

[0019] In another preferred embodiment,

[0020] R1 is substituted or unsubstituted phenyl, wherein the substitution refers to substitution by one or more substituents selected from the group consisting of halogen, cyano, C 1-6 Alkyl, C 3-6Cycloalkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyloxy, halogenated C 1-6 Alkyl, halogenated C 3-6 Cycloalkyl, halogenated C 1-6 Alkoxy, halogenated C 3-6 Cycloalkyloxy, -NR8R9, ethynyl;

[0021] R2 and R3 are independently selected from the group consisting of hydrogen, deuterium;

[0022] R4 and R5 are methyl groups;

[0023] R6 is selected from the following substituted or unsubstituted groups: C 1-6 Alkyl, C 3-6 Cycloalkyl, the substitution refers to substitution by one or more substituents selected from the group consisting of halogen, C 1-6 alkyl;

[0024] n is 1;

[0025] W is C-R7;

[0026] R7 is selected from the group consisting of hydrogen, halogen;

[0027] R8 and R9 are independently C 1-6 alkyl.

[0028] In another preferred embodiment,

[0029] R1 is substituted or unsubstituted phenyl, wherein the substitution refers to substitution by one or more substituents selected from the group consisting of halogen, cyano, C 1-6 Alkyl, C 3-6 Cycloalkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyloxy, halogenated C 1-6 Alkyl, halogenated C 3-6 Cycloalkyl, halogenated C 1-6 Alkoxy, halogenated C 3-6 Cycloalkyloxy, -NR8R9, ethynyl;

[0030] R2 and R3 are independently selected from the group consisting of hydrogen, deuterium;

[0031] R4 and R5 are methyl groups;

[0032] R6 is selected from the group consisting of:

[0033] n is 1;

[0034] W is C-R7;

[0035] R7 is selected from the group consisting of hydrogen, halogen;

[0036] R8 and R9 are independently C 1-6 alkyl.

[0037] In another preferred embodiment,

[0038] R1 is substituted or unsubstituted phenyl, wherein the substitution refers to substitution by one or more substituents selected from the group consisting of halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, -NR8R9, ethynyl;

[0039] R2 and R3 are independently selected from the group consisting of hydrogen, deuterium;

[0040] R4 and R5 are methyl groups;

[0041] R6 is selected from the group consisting of:

[0042] n is 1;

[0043] W is C-R7;

[0044] R7 is selected from the group consisting of hydrogen, halogen;

[0045] R8 and R9 are independently C 1-6 alkyl.

[0046] In another preferred embodiment, the compound is selected from the following group:

[0047]

[0048] The second aspect of the present invention provides a method for preparing the compound or a pharmaceutically acceptable salt thereof according to the first aspect of the present invention, comprising the steps of:

[0049]

[0050] in:

[0051] X is selected from the group consisting of halogen, -B(OH)2, -OTf;

[0052] R1, R2, R3, R4, R5, R6, n and W are as defined in the first aspect of the present invention.

[0053] The third aspect of the present invention provides a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers and a therapeutically effective amount of one or more compounds described in the first aspect of the present invention or a pharmaceutically acceptable salt thereof.

[0054] The fourth aspect of the present invention provides a use of the compound or a pharmaceutically acceptable salt thereof according to the first aspect of the present invention for preparing a medicament for preventing and / or treating diseases sensitive to potassium ion channels.

[0055] In another preferred embodiment, the disease sensitive to potassium ion channels is a central nervous system disease.

[0056] In another preferred embodiment, the central nervous system disease is selected from the group consisting of epilepsy, convulsions, inflammatory pain, neuropathic pain, migraine, depression, anxiety disorders, stroke, Alzheimer's disease, neurodegenerative diseases, cocaine abuse, nicotine withdrawal, alcohol withdrawal, and tinnitus.

[0057] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. DETAILED DESCRIPTION

[0058] After long-term and in-depth research, the inventor unexpectedly prepared a compound represented by formula A with excellent potassium channel opening activity, pharmacokinetics (such as brain-to-blood ratio performance, etc.), in vivo efficacy and safety and novel structure through structural optimization. On this basis, the inventor completed the present invention.

[0059] the term

[0060] In the present invention, unless otherwise specified, the terms used have the general meanings well known to those skilled in the art.

[0061] In the present invention, the term "halogen" refers to F, Cl, Br or I.

[0062] In the present invention, "C1-C6 alkyl" refers to a straight or branched alkyl group comprising 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, neopentyl, tert-pentyl, or the like.

[0063] In the present invention, the term "C2-C6 alkenyl" refers to a straight or branched alkenyl group having 2 to 6 carbon atoms and containing one double bond, including but not limited to ethenyl, propenyl, butenyl, isobutenyl, pentenyl and hexenyl.

[0064] In the present invention, the term "C2-C6 alkynyl" refers to a straight or branched alkynyl group having 2 to 6 carbon atoms and containing one triple bond, including but not limited to ethynyl, propynyl, butynyl, isobutynyl, pentynyl and hexynyl.

[0065] In the present invention, the term "C3-C8 cycloalkyl" refers to a cyclic alkyl group having 3-8 carbon atoms in the ring, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, etc. The terms "C3-C6 cycloalkyl" and "C3-C10 cycloalkyl" have similar meanings.

[0066] In the present invention, the term "C1-C6 alkoxy" refers to a straight or branched alkoxy group having 1 to 6 carbon atoms, including but not limited to methoxy, ethoxy, propoxy, isopropoxy and butoxy, etc. Preferably, it is a C1-C4 alkoxy group.

[0067] In the present invention, the term "aromatic ring" or "aryl" has the same meaning, preferably "C6-C10 aryl". The term "C6-C10 aryl" refers to an aromatic ring group having 6 to 10 carbon atoms without heteroatoms in the ring, such as phenyl, naphthyl, etc. The term "C 6-10 "C6-C10 aryl" and "C6-C10 aryl" have the same meaning, and other similar terms also have similar identity.

[0068] In the present invention, the term "aromatic heterocycle" or "heteroaryl" has the same meaning and refers to a heteroaromatic group containing one to multiple heteroatoms. For example, "C3-C10 heteroaryl" refers to an aromatic heterocycle containing 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen and 3 to 10 carbon atoms. Non-limiting examples include: furanyl, thienyl, pyridyl, pyrazolyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl ring can be fused to an aryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring. The heteroaryl group can be optionally substituted or unsubstituted.

[0069] In the present invention, the term "halo" means substituted with halogen.

[0070] In the present invention, the term "substituted" refers to one or more hydrogen atoms on a specific group being replaced by a specific substituent. The specific substituent is the substituent described above, or the substituent appearing in each embodiment. Unless otherwise specified, a substituted group may have a substituent selected from a specific group at any substitutable position of the group, and the substituent may be the same or different at each position. It should be understood by those skilled in the art that the combination of substituents contemplated by the present invention is those stable or chemically feasible combinations. The substituents include, for example (but not limited to): halogen, hydroxyl, carboxyl (-COOH), C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3- to 12-membered heterocyclic radical, aryl, heteroaryl, C1-C8 aldehyde, C2-C10 acyl, C2-C10 ester, amino, C1-C6 alkoxy, C1-C10 sulfonyl, etc.

[0071] In the present invention, the term multiple refers to 2, 3, 4, 5 or 6.

[0072] In the present invention, the term 1-6 means 1, 2, 3, 4, 5 or 6. Other similar terms have similar meanings.

[0073] Compound

[0074] The present invention provides a compound represented by formula A or a pharmaceutically acceptable salt thereof,

[0075]

[0076] R1, R2, R3, R4, R5, R6, n, and W are as defined above.

[0077] In another preferred embodiment, in the compound, any one of R1, R2, R3, R4, R5, R6, n, and W is a corresponding group in the specific compound.

[0078] As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by a compound of the present invention and an acid or base that is suitable for use as a drug. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is a salt formed by a compound of the present invention and an acid. Suitable acids for forming salts include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, and naphthalenesulfonic acid; and amino acids such as proline, phenylalanine, aspartic acid, and glutamic acid.

[0079] Another preferred salt is a salt of the compound of the present invention and a base, such as an alkali metal salt (e.g., sodium salt or potassium salt), an alkaline earth metal salt (e.g., magnesium salt or calcium salt), an ammonium salt (e.g., lower alkanolammonium salt and other pharmaceutically acceptable amine salts), for example, methylamine salt, ethylamine salt, propylamine salt, dimethylamine salt, trimethylamine salt, diethylamine salt, triethylamine salt, tert-butylamine salt, ethylenediamine salt, hydroxyethylamine salt, dihydroxyethylamine salt, trihydroxyethylamine salt, and amine salts formed from morpholine, piperazine, and lysine, respectively.

[0080] Preparation method

[0081] The preparation method of the compound of formula A of the present invention is described in more detail below, but these specific methods do not constitute any limitation to the present invention. The compound of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combination can be easily carried out by a technician in the field to which the present invention belongs.

[0082] Typically, the preparation process of the compound of the present invention is as follows, wherein the raw materials and reagents used can be purchased through commercial channels unless otherwise specified.

[0083]

[0084] in:

[0085] X is selected from the group consisting of halogen, -B(OH)2, -OTf;

[0086] R1, R2, R3, R4, R5, R6, n, and W are as defined above.

[0087] Pharmaceutical compositions and methods of administration

[0088] The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, and more preferably, contains 5-1000 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

[0089] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0090] The pharmaceutical composition is in the form of injection, capsule, tablet, pill, powder or granule.

[0091] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administrations include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0092] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0093] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0094] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

[0095] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0096] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances.

[0097] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0098] Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0099] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

[0100] The treatment method of the present invention can be used alone or in combination with other treatment methods or therapeutic drugs.

[0101] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 5 to 1000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill range of a skilled physician.

[0102] Compared with the prior art, the present invention has the following main advantages:

[0103] (1) The compound has better pharmacokinetic properties, such as better brain-to-blood ratio, half-life, exposure, metabolic stability, etc.;

[0104] (2) The compound has better potassium ion channel opening activity, better ion channel selectivity, better in vivo efficacy and better safety;

[0105] (3) The compounds are expected to be useful for treating and / or preventing diseases and disorders affected by the activity of potassium ion channels.

[0106] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. The experimental methods in the following examples where specific conditions are not specified are generally performed under conventional conditions such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or under conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0107] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0108] Unless otherwise specified, the experimental materials and reagents used in the following examples can be obtained from commercial channels.

[0109] Example 1 Preparation of Compound A

[0110]

[0111] Step 1: Compound 2

[0112] Compound 1 (2 g, 8.05 mmol, 1.0 eq) was dissolved in a mixed solvent of dichloromethane (100 mL) and methanol (10 mL), and then BnMe3NBr3 (6.28 g, 16.1 mmol, 2.0 eq) and calcium carbonate (2.01 g, 20.1 mmol, 2.5 eq) were added, and the reaction solution was stirred at room temperature for 0.5 hours. The reaction solution was filtered, and the residue obtained after the filtrate was concentrated was purified by neutral alumina column chromatography (petroleum ether: ethyl acetate = 15: 1) to obtain compound 2 (2.9 g, yield 90%) as a white solid.

[0113] LCMS: [M+H] + =406.9.

[0114] Step 2: Compound 3

[0115] Compound 2 (1 g, 2.46 mmol, 1.0 eq) was dissolved in 1,4-dioxane (50 mL), and then methylboric acid (589 mg, 9.84 mmol, 4.0 eq), potassium phosphate (2.089 g, 9.84 mmol, 4.0 eq) and Pd(dppf)Cl2 (0.1 g, 0.14 mmol, 0.06 eq) were added. The reaction solution was heated to 120 ° C and stirred for 4 h under nitrogen protection. After cooling to room temperature, the reaction solution was filtered, the filtrate was diluted with water, and then extracted with ethyl acetate (3×40 mL), the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the residue obtained after concentration was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to obtain compound 3 (504 mg, yield 74%) as a white solid.

[0116] LCMS: [M+H] + =277.2.

[0117] Step 3: Compound 5

[0118] Compound 3 (600 mg, 2.17 mmol, 1.0 eq) was dissolved in dichloromethane (15 mL), cooled to 0 ° C, triethylamine (0.78 mL, 5.43 mmol, 2.5 eq) and compound 4 (438 mg, 3.26 mmol, 1.5 eq) were added, and the reaction solution was warmed to room temperature and stirred for 0.5 hours. The reaction solution was diluted with water, then extracted with dichloromethane (3×10 mL), the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and concentrated, and the residue was purified by slurrying with a mixed solvent (petroleum ether / ethyl acetate = 30 mL / 3 mL) for half an hour and filtered. The obtained solid was dried to obtain compound 5 (680 mg, yield 84%) as a white solid.

[0119] Step 4: Compound 6

[0120] Compound 5 (680 mg, 1.82 mmol, 1.0 eq) and methanol solution of hydrochloric acid (4 M, 13 mL) were added to a single-mouth bottle and stirred at room temperature for 0.5 hours. The reaction solution was concentrated and diluted with ethyl acetate (10 mL), then washed with sodium carbonate aqueous solution (0.5 M, 20 mL), and the separated aqueous phase was extracted with ethyl acetate (3×10 mL). The combined organic phase was washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The organic layer was concentrated and the residue was dissolved in tetrahydrofuran, dried over anhydrous sodium sulfate, and concentrated to obtain compound 6 (370 mg, yield 74%) as a white solid.

[0121] LCMS: [M+H] + =275.2

[0122] Step 5: Compound A

[0123] Compound 6 (150 mg, 0.547 mmol, 1.5 eq) was dissolved in toluene (50 mL), and then cesium carbonate (237 mg, 0.728 mmol, 2.0 eq), X-Phos (35 mg, 0.0728 mmol, 0.2 eq), compound 7 (81 mg, 0.364 mmol, 1.0 eq) and Pd2(dba)3 (34 mg, 0.0364 mmol, 0.1 eq) were added in sequence, and the reaction solution was stirred at 90°C for 16 hours under nitrogen protection. The reaction solution was cooled to room temperature and filtered, and the filtrate was diluted with water and then extracted with ethyl acetate (3×20 mL). The organic phase was washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The residue obtained after concentration was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain compound A (6.0 mg, 3%) as a white solid.

[0124] LCMS: [M+H] + =369.2

[0125] 1 H NMR(400MHz,DMSO)δ9.10(s,1H),7.08-7.02(m,4H),6.90(s,1H),4.26(s,2H),3. 49(s,2H),2.74-2.71(m,2H),2.21(s,2H),2.11(s,3H),2.01(s,3H),1.06(s,9H).

[0126] Example 2 Preparation of Compound B

[0127]

[0128] Step 1: Compound B

[0129] Compound 1 (600 mg, 2.19 mmol, 1.0 eq), compound 2 (3.6 g, 27.3 mmol, 12.4 eq) and cesium carbonate (7.14 g, 21.9 mmol, 10 eq) were added to N-methylpyrrolidone (60 mL), and the reaction solution was heated to 180 ° C in a microwave reactor and stirred for 4 hours. The reaction solution was cooled to room temperature and diluted with ethyl acetate (150 mL), then washed with saturated sodium chloride solution (80 mL x 4), dried over anhydrous sodium sulfate and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to obtain compound B (83.9 mg, yield 10%) as a white solid.

[0130] LCMS: [M+H] + =387.1

[0131] 1 H NMR(400MHz,DMSO-d6)δ9.11(s,1H),7.21-7.15(m,1H),6.94-6.84(m,2H),6.80-6.69(m,1H),4.19 (s,2H),3.44-3.36(m,2H),2.76-2.71(m,2H),2.21(s,2H),2.10(s,3H),2.01(s,3H),1.05(s,9H).

[0132] Example 3 Preparation of Compound C

[0133]

[0134] Step 1: Compound C

[0135] Compound 1 (100 mg, 0.36 mmol, 1.0 eq) was dissolved in N-methylpyrrolidone (5 mL), and then cesium carbonate (1.0 g, 3.1 mmol, 8.6 eq) and compound 2 (1.0 g, 6.67 mmol, 18.5 eq) were added. The reaction solution was heated to 200 ° C in a microwave reactor and stirred for 2 hours, cooled to room temperature and filtered, the solid was washed with ethyl acetate (3 x 5 mL), and the filtrate was washed with saturated sodium chloride solution (3 x 10 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1) to obtain compound C (15.5 mg, yield 10%) as a white solid.

[0136] LCMS: [M+H] + =405.1

[0137] 1 H NMR(400MHz,DMSO-d6)δ9.13(s,1H),7.58-7.46(m,1H),7.23-7.16(m,1H),6.86(s,1H),4.15(s ,2H),3.37-3.33(m,2H),2.79-2.72(m,2H),2.22(s,2H),2.11(s,3H),2.02(s,3H),1.07(s,9H).

[0138] Example 4 Preparation of Compound D

[0139]

[0140] Step 1: Compound 3

[0141] Compound 1 (50 mg, 0.182 mmol, 1.0 eq) was dissolved in tetrahydrofuran (5 mL), and then compound 2 (164 mg, 0.547 mmol, 3.0 eq), Pd2(dba)3 (17 mg, 0.018 mmol, 0.1 eq), Dave-Phos (14 mg, 0.036 mmol, 0.2 eq) and LiHMDS (1 M, 1.8 mL, 1.8 mmol, 10 eq) were added in sequence. The reaction solution was stirred at 80 ° C for 2 hours under nitrogen protection, cooled to room temperature, diluted with ethyl acetate (150 mL), and then washed with saturated sodium chloride solution (80 mL x 4). The organic phase was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain compound 3 (46 mg, yield 28%) as a yellow solid.

[0142] LCMS: [M+H] + =447.2

[0143] Step 2: Compound D

[0144] Compound 3 (20 mg, 0.045 mmol, 1.0 eq) and potassium carbonate (62 mg, 0.447 mmol, 10 eq) were added to methanol (3 mL), and the reaction solution was stirred at 25° C. for 2 hours. The reaction solution was concentrated, and the obtained residue was dissolved in ethyl acetate (20 mL), and then washed with saturated sodium chloride solution (10 mL x 2), and the obtained organic phase was dried over anhydrous sodium sulfate and concentrated, and the residue was purified by pre-HPLC (0.1% formic acid / acetonitrile / water) to obtain compound D (4.2 mg, yield 7%) as a light red solid.

[0145] LCMS: [M+H] + =375.2

[0146] 1 H NMR (400MHz, DMSO-d6) δ9.11 (s, 1H), 7.31 (d, J = 8.8Hz, 2H), 7.03-6.88 (m, 3H), 4.38 (s, 2H), 3. 90(s,1H),3.61(s,2H),2.75-2.72(m,2H),2.21(s,2H),2.10(s,3H),2.02(s,3H),1.06(s,9H).

[0147] Example 5 Preparation of Compound E

[0148]

[0149] Step 1: Compound E

[0150] Compound 1 (100 mg, 0.36 mmol, 1.0 eq) was dissolved in dimethyl sulfoxide (2 mL), and then compound 2 (66 mg, 0.55 mmol, 1.5 eq) and potassium carbonate (151 mg, 1.09 mmol, 3.0 eq) were added in sequence, and the reaction solution was stirred at 100 ° C for 16 hours. After cooling to room temperature, water (20 mL) was added to the reaction solution, and extracted with ethyl acetate (3 x 20 mL). The combined organic phase was washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The concentrated residue was purified by silica gel column chromatography (dichloromethane / methanol = 50 / 1) to obtain compound E (50.3 mg, yield 37%) as a white solid.

[0151] LCMS: [M+H] + =376.2

[0152] 1H NMR(400MHz,DMSO-d6)δ9.13(s,1H),7.60-7.57(m,2H),7.08-7.05(m,2H),6.94(s,1H),4.49( s,2H),3.69(s,2H),2.77(t,J=5.6Hz,2H),2.21(s,2H),2.11(s,3H),2.03(s,3H),1.06(s,9H).

[0153] Example 6 Preparation of Compound F

[0154]

[0155] Step 1: Compound F

[0156] Compound 1 (120 mg, 0.44 mmol, 1.0 eq) was dissolved in acetonitrile (20 mL), and compound 2 (148 mg, 1.31 mmol, 3.0 eq) and diisopropylethylamine (339 mg, 2.63 mmol, 6.0 eq) were added, and the mixture was heated to 160 ° C in a microwave reactor for 1 hour. The reaction solution was cooled to room temperature and concentrated, diluted with water, and then extracted with dichloromethane (3 x 100 mL). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to obtain compound F (40 mg, yield 26%) as a yellow solid.

[0157] LCMS: [M+H] + =352.2

[0158] Example 7 Preparation of Compound G

[0159]

[0160] Step 1: Compound 3

[0161] Compound 1 (600 mg, 3.0 mmol, 1.0 eq) and compound 2 (780 mg, 3.9 mmol, 1.3 eq) were dissolved in ammonia methanol solution (1 M, 10.5 mL), and the reaction solution was heated to 90 ° C in a microwave reactor for half an hour. The reaction was also cooled to room temperature and concentrated, extracted with ethyl acetate (50 mL x 3) and saturated sodium bicarbonate aqueous solution, the organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 100 / 15) to obtain compound 3 (350 mg, yield 42%) as a white solid.

[0162] Step 2: Compound 4

[0163] Compound 3 (350 mg, 1.25 mmol, 1.0 eq) and palladium carbon (10%, 100 mg) were dissolved in methanol (30 mL) and stirred at room temperature for 1 hour under 1 atmosphere of hydrogen. The reaction solution was filtered and the filtrate was concentrated to obtain compound 4 (300 mg, yield 96%) as a colorless oil.

[0164] LCMS: [M+H] + =250.1.

[0165] Step 3: Compound 5

[0166] Compound 4 (300 mg, 1.2 mmol, 1.0 eq), benzyltrimethylammonium tribromide (936 mg, 2.4 mmol, 2.0 eq) and calcium carbonate (300 mg, 3.0 mmol, 2.5 eq) were dissolved in a mixed solvent of dichloromethane / methanol (10 / 1, 40 mL / 4 mL) and stirred at room temperature for half an hour. The reaction solution was filtered, and the filtrate was concentrated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to obtain compound 5 (160 mg, yield 33%) as a white solid.

[0167] LCMS: [M+Na] + =429.9.

[0168] Step 4: Compound 7

[0169] Compound 5 (160 mg, 0.4 mmol, 1.0 eq) was dissolved in 1,4-dioxane (15 mL), and methylboric acid (96 mg, 1.6 mmol, 4.0 eq), Pd(dppf)Cl2 (58 mg, 0.08 mmol, 0.2 eq) and potassium phosphate (339 mg, 1.6 mmol, 4.0 eq) were added. The reaction solution was heated to 120 ° C under a nitrogen atmosphere for 4 hours. The reaction solution was cooled to room temperature and filtered, and the residue obtained after the filtrate was concentrated was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 65 / 35) to obtain compound 7 (80 mg, yield 72%) as a white solid.

[0170] LCMS: [M+H] + =278.1

[0171] Step 5: Compound 9

[0172] Compound 7 (80 mg, 0.3 mmol, 1.0 eq) and triethylamine (116 mg, 0.9 mmol, 3.0 eq) were dissolved in dichloromethane (15 mL), and compound 8 (60 mg, 0.45 mmol, 1.5 eq) was slowly added under nitrogen atmosphere, and stirred at room temperature for 1 hour. The reaction solution was extracted with ethyl acetate (50 mL x 3) and saturated sodium bicarbonate aqueous solution, and the combined organic phase was dried over anhydrous sodium sulfate and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 65 / 35) to obtain compound 9 (80 mg, 71%) as a light yellow solid.

[0173] LCMS: [M+H] + =376.3

[0174] Step 6: Compound 10

[0175] Compound 9 (80 mg, 0.21 mmol, 1.0 eq) was dissolved in a methanol solution of hydrogen chloride (4 M, 4 mL) and stirred at room temperature for 1 hour. The reaction solution was concentrated to obtain compound 10 (70 mg, yield 96%) as a light yellow oil.

[0176] LCMS: [M+H] + =276.2

[0177] Step 7: Compound G

[0178] Compound 10 (30 mg, 0.11 mmol, 1.0 eq) and compound 11 (38 mg, 0.17 mmol, 1.5 eq) were dissolved in toluene (10 mL), and then Pd2(dba)3 (20 mg, 0.022 mmol, 0.2 eq), X-Phos (10 mg, 0.022 mmol, 0.2 eq) and cesium carbonate (90 mg, 0.275 mmol, 2.5 eq) were added in sequence, and heated to 100°C under a nitrogen atmosphere for 16 hours. The reaction solution was cooled to room temperature and concentrated, and the obtained residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain compound G (5.3 mg, yield 13%) as a white solid.

[0179] LCMS: [M+H] + =370.1

[0180] 1H NMR (400MHz, CDCl3) δ7.02-6.94(m,4H),6.80(s,1H),4.35(s,2H),3.50(t,J=5.6H z,2H),2.84(t,J=5.6Hz,2H),2.48(s,3H),2.33(s,2H),2.15(s,3H),1.16(s,9H).

[0181] Example 8 Preparation of Compound H

[0182]

[0183] Step 1: Compound 3

[0184] Compound 1 (5.0 g, 24.63 mmol, 1.0 eq) was dissolved in toluene (100 mL), and compound 2 (3.88 g, 36.945 mmol, 1.5 eq) was added, and the reaction solution was heated to 140°C and stirred for 16 hours. After cooling to room temperature, the reaction solution was concentrated to obtain compound 3 (7 g, yield 98%) as a brown oil, which was directly used in the next step without purification.

[0185] Step 2: Compound 4

[0186] Compound 3 (3 g, 10.344 mmol, 1.0 eq) was dissolved in methanol (30 mL), cooled to 0 ° C, sodium borohydride (0.27 g, 7.241 mmol, 0.7 eq) was added, and the mixture was heated to room temperature and stirred for 2 hours. Ice water was added to the reaction solution, and the methanol was concentrated to remove the methanol, and ethyl acetate (3 x 30 mL) was added for extraction. The combined organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the residue obtained after concentration was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4 / 1) to obtain compound 4 (2.5 g, yield 82%) as a white solid.

[0187] LCMS: [M+H] + =292.0

[0188] Step 3: Compound 5

[0189] Compound 4 (7.45 g, 25.5 mmol, 1.0 eq) was dissolved in dichloromethane (200 mL), and 4-dimethylaminopyridine (156 mg, 1.28 mmol, 0.05 eq) and triethylamine (5.16 g, 51.0 mmol, 2.0 eq) were added in sequence. The reaction solution was cooled to 0 °C under nitrogen protection, and p-toluenesulfonyl chloride (5.10 g, 26.8 mmol, 1.05 eq) was added. The reaction solution was heated to room temperature and stirred for 16 hours, then quenched with ice water (200 mL), and the aqueous phase was extracted with dichloromethane (200 mL). The separated organic phase was dried over anhydrous sodium sulfate and concentrated, and the obtained residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to obtain compound 5 (11.0 g, yield 96%).

[0190] LCMS: [M+Na] + =469.9

[0191] Step 4: Compound 6

[0192] Dichloromethane (300 mL) was added to a reaction bottle containing aluminum chloride (22.0 g, 165 mmol, 6.7 eq), cooled to 0 ° C under nitrogen protection, and a dichloromethane (100 vmL) solution of compound 5 (11.0 g, 24.65 mmol, 1.0 eq) was added. The reaction solution was heated to room temperature and stirred for 16 hours, then quenched with ice water (100 mL), the pH value of the solution was adjusted to 10 with ammonia water, and the aqueous phase was extracted with dichloromethane (200 mL). The combined organic phase was dried over anhydrous sodium sulfate, and the residue obtained after concentration was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4 / 1) to obtain compound 6 (2.6 g, yield 47%).

[0193] LCMS: [M+H] + =225.9

[0194] Step 5: Compound 7

[0195] Compound 6 (2.6 g, 11.5 mmol, 1.0 eq) was dissolved in acetic acid (40 mL), cooled to 0 ° C under nitrogen protection, and sodium borohydride (1.3 g, 34.5 mmol, 3.0 eq) was added. The reaction solution was heated to room temperature and stirred for 2 hours, diluted with water (200 mL), cooled to 0 ° C, and the pH value of the solution was adjusted to 10 with sodium carbonate, and then extracted with ethyl acetate (3 x 200 mL). The combined organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain crude compound 7 (3.0 g), which was directly used in the next step without purification.

[0196] LCMS: [M+H] + =230.0

[0197] Step 6: Compound 8

[0198] Compound 7 (3.0 g, 11.5 mmol, 1.0 eq) was dissolved in tetrahydrofuran (40 mL), and diisopropylethylamine (743 mg, 5.75 mmol, 0.5 eq) and Boc2O (3.0 g, 13.8 mmol, 1.2 eq) were added. The reaction solution was stirred at room temperature for 16 hours and concentrated. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to obtain compound 8 (3.0 g, two-step yield 79%).

[0199] LCMS: [M+Na] + =352.0

[0200] Step 7: Compound 10

[0201] Compound 8 (3.0 g, 9.09 mmol, 1.0 eq) and compound 9 (3.3 g, 18.7 mmol, 2.0 eq) were dissolved in toluene (100 mL), and then cesium carbonate (8.88 g, 27.26 mmol, 3.0 eq), BINAP (1.13 g, 1.82 mmol, 0.2 eq) and Pd2(dba)3 (832 mg, 0.09 mmol, 0.1 eq) were added under nitrogen protection. The reaction solution was heated to 80 ° C and stirred for 20 hours, and then heated to 100 ° C and stirred for 2 hours. After cooling to room temperature, the reaction solution was filtered, and the solid was washed with ethyl acetate (200 mL). The combined filtrate was washed with water and saturated sodium chloride solution, and then dried over anhydrous sodium sulfate and concentrated to obtain crude compound 10 (6.0 g), which was directly used in the next step without purification.

[0202] LCMS: [M+H] + =431.1

[0203] Step 8: Compound 11

[0204] Compound 10 (crude product, 6.0 g, 9.09 mmol, 1.0 eq) was dissolved in dichloromethane (200 mL), and aqueous hydrochloric acid solution (4 M, 100 mL) was added and stirred at room temperature for 2 hours. The reaction solution was separated, the organic phase was washed with aqueous hydrochloric acid solution (4 M, 3 x 30 mL), and the combined aqueous phase was adjusted to pH 8 with sodium carbonate to obtain an aqueous solution of compound 11 (about 200 mL).

[0205] LCMS: [M+H] + =167.1

[0206] Step 9: Compound 12

[0207] A solution of Boc2O (4.0 g, 18 mmol, 2.0 eq) in tetrahydrofuran (100 mL) was added to the aqueous solution of compound 11 (about 200 mL), and the mixture was stirred at room temperature for 4 hours. The reaction solution was extracted with ethyl acetate (3 x 50 mL), and the combined organic phase was washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the residue obtained after concentration was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to obtain compound 12 (3.0 g, three-step yield 90%).

[0208] LCMS: [M+Na] + =389.1

[0209] Step 10, Compound 13

[0210] Compound 12 (2.8 g, 7.65 mmol, 1.0 eq) was dissolved in a methanol solution of hydrogen chloride (4 M, 40 mL) and stirred at room temperature for 2 hours. The reaction solution was concentrated to obtain compound 13 (2.0 g) as a white solid.

[0211] LCMS: [M+H] + =167.1

[0212] Step 11, Compound 14

[0213] Compound 13 (2.0 g) was dissolved in tetrahydrofuran (50 mL), and the pH value was adjusted to 8 with sodium carbonate aqueous solution, and Boc2O (1.5 g, 6.88 mmol, 0.9 eq) was added. The reaction solution was stirred at room temperature for 10 minutes and extracted with ethyl acetate (3×60 mL). The combined organic phase was washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The residue obtained after concentration was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain compound 14 (0.85 g, two-step yield 42%).

[0214] LCMS: [M+H] + =267.1

[0215] Step 12: Compound 15

[0216] Compound 14 (0.8 g, 3.0 mmol, 1.0 eq) was dissolved in a mixed solvent of dichloromethane (100 mL) and methanol (10 mL), calcium carbonate (2.1 g, 21.03 mmol, 7.0 eq) was added, and the temperature was lowered to 0 ° C under nitrogen protection. BnMe3NBr3 (4.5 g, 11.5 mmol, 4.0 eq) was slowly added to the reaction solution in batches, and the reaction solution was stirred at room temperature for half an hour. The reaction solution was filtered and the solid was washed with dichloromethane (100 mL). The combined filtrate was washed with a mixed aqueous solution of sodium bicarbonate and sodium sulfite, the organic phase was dried over anhydrous sodium sulfate and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to obtain compound 15 (1.1 g, yield 80%) as a white solid.

[0217] LCMS: [M+Na] + =446.8

[0218] Step 13, Compound 17

[0219] Compound 17 (1.0 g, 2.36 mmol, 1.0 eq) was dissolved in 1,4-dioxane (100 mL), methylboric acid (1.13 g, 18.86 mmol, 4.0 eq), potassium phosphate (2.5 g, 11.79 mmol, 5.0 eq) and Pd(dppf)Cl2 (289 mg, 0.35 mmol, 0.15 eq) were added, and the mixture was heated to 120 ° C and stirred for 8 hours under nitrogen protection. The reaction solution was cooled to room temperature and filtered, and the filtrate was concentrated and dissolved with water and ethyl acetate and separated, and the aqueous phase was extracted with ethyl acetate. The combined organic phase was washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The concentrated residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 6 / 1) to obtain compound 17 (650 mg, yield 85%) as a white solid.

[0220] LCMS: [M+H] + =295.1

[0221] Step 14: Compound 19

[0222] Compound 17 (600 mg, 2.04 mmol, 1.0 eq) was dissolved in dichloromethane (20 mL), cooled to 0 ° C, and triethylamine (515 mg, 5.10 mmol, 2.5 eq) and compound 18 (411 mg, 3.06 mmol, 1.5 eq) were added. The reaction solution was heated to room temperature and stirred for 0.5 hours, the reaction solution was diluted with water, and then extracted with dichloromethane (3×50 mL). The combined organic phase was washed with water, dried over anhydrous sodium sulfate, and the residue after concentration was slurried with a mixed solvent (petroleum ether / ethyl acetate = 40 / 1) for half an hour. The solid obtained after filtration was dried to obtain compound 19 (700 mg, yield 87%) as a white solid.

[0223] LCMS: [M+Na] + =415.2

[0224] Step 15: Compound 20

[0225] Compound 19 (700 mg, 1.78 mmol, 1.0 eq) and a methanol solution of hydrogen chloride (4 M, 15 mL) were added to a single-mouth bottle and stirred at room temperature for 0.5 hours. The reaction solution was spin-dried and diluted with ethyl acetate, and then the pH value was adjusted to 8 with an aqueous sodium carbonate solution, and the aqueous phase was extracted with ethyl acetate. The combined organic phase was washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain compound 20 (400 mg, 77%).

[0226] LCMS: [M+H] + =293.2

[0227] Step 16: Compound H

[0228] Compound 20 (180 mg, 0.61 mmol, 1.0 eq), compound 21 (1.08 g, 8.18 mmol, 13.0 eq) and cesium carbonate (2.0 g, 6.1 mmol, 10 eq) were added to N-methylpyrrolidone (10 mL), and heated to 180° C. in a microwave reactor under nitrogen for 4 hours. The reaction solution was cooled to room temperature and diluted with ethyl acetate (150 mL), and then washed with saturated sodium chloride solution (80 mL x 4). The combined organic phase was dried over anhydrous sodium sulfate, and the residue obtained after concentration was purified by silica gel column chromatography (petroleum ether / ethyl acetate=30 / 1 to 2 / 1) to obtain compound H (4.2 mg, yield 17%) as a white solid.

[0229] LCMS: [M+Na] + =405.1

[0230] 1H NMR(400MHz,DMSO-d6)δ9.30(s,1H),7.21-7.18(m,1H),7.00-6.96(m,1H),6.82-6.77(m,1H),4.23 (s,2H),3.44-3.36(m,2H),2.82-2.73(m,2H),2.24(s,2H),2.04(s,3H),2.01(s,3H),1.07(s,9H).

[0231] Example 9 Preparation of Compound I

[0232]

[0233] Step 1: Compound 5

[0234] Compound 3 (1.2 g, 10.8 mmol, 1.5 eq) was dissolved in anhydrous dimethylformamide (5 mL), and compound 4 (2.0 g, 7.2 mmol, 1.0 eq), HATU (6.84 g, 18 mmol, 2.5 eq) and diisopropylethylamine (6 mL, 36 mmol, 5.0 eq) were added in sequence, and the reaction solution was heated to 45 ° C and stirred for 16 hours. After the reaction solution was cooled to room temperature, it was diluted with water and extracted with ethyl acetate (3×10 mL), and the combined organic phase was washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The residue obtained after concentration was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1) to obtain compound 5 (2.5 g, yield 62%) as a white solid.

[0235] LCMS: [M+Na] + =395.2.

[0236] Step 2: Compound 6

[0237] Compound 5 (2.5 g, 6.7 mmol, 1.0 eq) and a methanol solution of hydrogen chloride (4 M, 55 mL) were added to a single-mouth bottle in sequence and stirred at room temperature for 1 hour. The reaction solution was concentrated, and the residue was dissolved in a mixed solvent of dichloromethane / methanol (10 / 1), then washed with an aqueous sodium carbonate solution (0.5 M, 50 mL), and the aqueous phase was extracted with a mixed solvent of dichloromethane / methanol (10 / 1, 3×20 mL). The combined organic phase was washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain compound 6 (1.6 g, yield 85%) as a gray solid.

[0238] LCMS: [M+H] + =273.2

[0239] Step 3: Compound I

[0240] Compound 6 (1.4 g, 5.1 mmol, 1.0 eq) was dissolved in toluene (200 mL), and cesium carbonate (3.3 g, 10.2 mmol, 2.0 eq), compound 7 (1.7 g, 7.7 mmol, 1.5 eq), X-Phos (486 mg, 1.02 mmol, 0.2 eq) and Pd2(dba)3 (467 mg, 0.51 mmol, 0.1 eq) were added in sequence. The temperature was raised to 100 °C and stirred for 16 hours under nitrogen protection. The reaction solution was cooled to room temperature and filtered. The filtrate was diluted with water and extracted with ethyl acetate (3×20 mL). The combined organic phase was washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The concentrated residue was purified by neutral alumina column chromatography (petroleum ether / ethyl acetate=5 / 1) to obtain compound I. The crude compound I was further purified by Pre-HPLC (0.1% formic acid / acetonitrile / water) to obtain compound I (377.0 mg, 20%) as a white solid.

[0241] LCMS: [M+H] + =367.2

[0242] 1 H NMR(400MHz,DMSO-d6)δ9.02(s,1H),7.06-7.03(m,4H),6.91(s,1H),4.26(s,2H),3.50(s,2H),2.74-2 .72(m,2H),2.21(s,2H),2.11(s,3H),2.01(s,3H),1.15(s,3H),0.56-0.53(m,2H),0.33-0.31(m,2H).

[0243] Example 10 Preparation of Compound J

[0244]

[0245] Step 1: Compound J

[0246] Compound 1 (600 mg, 2.20 mmol, 1.0 eq), compound 2 (3.6 g, 27.3 mmol, 12.4 eq) and cesium carbonate (7.19 g, 22.0 mmol, 10 eq) were added to N-methylpyrrolidone (60 mL), and the reaction mixture was heated to 180 ° C in a microwave reactor and stirred for 4 hours. After cooling to room temperature, the reaction solution was diluted with ethyl acetate (100 mL), then washed with saturated sodium chloride solution (80 mL x 2), dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to obtain compound J (56.0 mg, 0.146 mmol, yield 7%) as a white solid.

[0247] LCMS: [M+H] + =385.1

[0248] 1 H NMR(400MHz, DMSO-d6)δ9.01(s,1H),7.19-7.12(m,1H),6.92-6.82(m,2H),6.75-6.67(m,1H),4.16(s,2H),3.37(t,J=5.6 Hz,2H),2.71(t,J=5.6Hz,2H),2.18(s,2H),2.07(s,3H),1.97(s,3H),1.11(s,3H),0.52-0.50(m,2H),0.29-0.27(m,2H).

[0249] Example 11 Preparation of Compound K

[0250]

[0251] Step 1: Compound 3

[0252] Compound 1 (200 mg, 0.724 mmol, 1.0 eq) was dissolved in dimethylformamide (5 ml), and compound 2 (128 mg, 1.086 mmol, 1.5 eq), HATU (412 mg, 1.086 mmol, 1.5 eq) and diisopropylethylamine (280 mg, 2.172 mmol, 3.0 eq) were added, and the mixture was heated to 60 ° C and stirred for 4 hours. The reaction solution was cooled to room temperature and diluted with water (20 mL), and the aqueous phase was extracted with ethyl acetate (3×20 mL). The combined organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the residue obtained after concentration was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain compound 3 (220 mg, yield 83%) as a yellow solid.

[0253] LCMS: [M+H] + =377.2

[0254] Step 2: Compound 4

[0255] Compound 3 (280 mg, 0.745 mmol, 1.0 eq) was dissolved in a methanol solution of hydrogen chloride (4 M, 10 mL) and stirred at room temperature for 1 hour. After concentration, a saturated aqueous sodium carbonate solution was added to the residue to adjust the pH value to 8-9, and extracted with ethyl acetate (3 x 30 mL). The combined organic phases were washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain compound 4 (180 mg, yield 88%) as a yellow solid.

[0256] LCMS: [M+H]+ =277.2

[0257] Step 3: Compound K

[0258] Compound 4 (10 mg, 0.036 mmol, 1.0 eq) was dissolved in N-methylpyrrolidone (1 mL), cesium carbonate (117 mg, 0.36 mmol, 10.0 eq) and compound 5 (58 mg, 0.446 mmol, 12.4 eq) were added, and the mixture was heated to 120 °C using a microwave reactor for 0.5 h. The reaction solution was cooled to room temperature and concentrated, and the residue was purified by prep-HPLC to obtain compound K.

[0259] LCMS: [M+H] + =389.1

[0260] Example 12 Preparation of Compound L

[0261]

[0262] Step 1: Compound 3

[0263] Compound 1 (100 mg, 0.54 mmol, 1.0 eq) was dissolved in dimethylformamide (3 mL), and compound 2 (81 mg, 0.81 mmol, 1.5 eq), HATU (314 mg, 0.81 mmol, 1.5 eq) and diisopropylethylamine (209 mg, 1.62 mmol, 3.0 eq) were added in sequence, and the mixture was reacted at room temperature for 1 hour. The reaction solution was extracted with ethyl acetate (50 mL x 3) and saturated sodium chloride solution, and the combined organic phase was dried over anhydrous sodium sulfate. The residue obtained after concentration was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 2) to obtain compound 3 (130 mg, yield 67%) as a white solid.

[0264] LCMS: [M+Na] + =381.1.

[0265] Step 2: Compound 4

[0266] Compound 3 (130 mg, 0.36 mmol, 1.0 eq) was dissolved in dichloromethane (10 mL), trifluoroacetic acid (2 mL) was added, and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated, extracted with ethyl acetate and saturated sodium bicarbonate aqueous solution, the organic phase was dried over anhydrous sodium sulfate, and concentrated to give compound 4 (80 mg, yield 86%) as a light yellow oil.

[0267] LCMS: [M+H] + =259.1

[0268] Step 3: Compound L

[0269] Compound 4 (80 mg, 0.31 mmol, 1.0 eq) and compound 5 (103 mg, 0.47 mmol, 1.5 eq) were dissolved in toluene (10 mL), Pd2(dba)3 (57 mg, 0.062 mmol, 0.2 eq), x-Phos (30 mg, 0.062 mmol, 0.2 eq) and cesium carbonate (253 mg, 0.78 mmol, 2.5 eq) were added, and the reaction solution was heated to 100 ° C under nitrogen protection for 18 hours. The reaction solution was cooled to room temperature and concentrated, and the obtained residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain compound L (8.8 mg, yield 8%) as a white solid.

[0270] LCMS: [M+H] + =353.1

[0271] 1 H NMR(400MHz, CDCl3)δ7.23(s,1H),7.02-6.93(m,4H),6.90(s,1H),4.27(s,2H),3.50-3.47(m,2H),2.84-2.82(m ,2H),2.39(d,J=7.6Hz,2H),2.23(s,3H),2.13(s,3H),1.19-1.11(m,1H),0.77-0.67(m,2H),0.36-0.33(m,2H).

[0272] Example 13 Preparation of Compound M

[0273]

[0274] Step 1: Compound 3

[0275] Compound 1 (6.0 g, 49.5 mmol, 1.0 eq) and diisopropylethylamine (12.8 g, 99.0 mmol, 2.0 eq) were added to dichloromethane (100 mL), and the above solution was cooled to 0°C under nitrogen protection, and compound 2 (5.83 g, 74.3 mmol, 1.5 eq) was slowly added dropwise to the reaction solution, and the reaction solution was reacted at room temperature for 2 hours under nitrogen protection. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1:1) to obtain a white solid 3 (6.0 g, yield 74%).

[0276] LCMS: [M+H] + =164.1.

[0277] Step 2: Compound 5

[0278] Aluminum chloride (9.6 g, 72.0 mmol, 3.0 eq) was added to dichloromethane (100 mL), and the reaction mixture was cooled to 0 ° C under nitrogen protection, and a dichloromethane (50 mL) solution of compound 3 (4.0 g, 24.0 mmol, 1.0 eq) and compound 4 (7.4 g, 36.6 mmol, 1.5 eq) was slowly added. The reaction solution was slowly heated to room temperature and stirred for 16 hours, diluted with dichloromethane (100 mL), and then washed with ice water (5 x 200 mL). The separated organic phase was dried over anhydrous sodium sulfate and concentrated to give a crude compound 5 (6.0 g). The crude compound 5 was slurried with a mixed solvent of dichloromethane / petroleum ether (20 mL / 40 mL) and filtered to give compound 5 (4.0 g, yield 58%) as a white solid.

[0279] LCMS: [M+H] + =284.0

[0280] Step 3: Compound 6

[0281] Compound 5 (2.0 g, 7.0 mmol, 1.0 eq) was dissolved in dichloromethane (10 mL), HSiEt3 (10 mL) was added, the reaction solution was cooled to 0 ° C under nitrogen protection, trifluoroacetic acid (10 mL) was slowly added, slowly warmed to room temperature and stirred for 16 hours. The above reaction solution was concentrated, the residue was dissolved in tetrahydrofuran (30 mL), then cooled to 0 ° C under nitrogen protection, sodium borohydride (532 mg, 14 mmol, 2.0 eq) was slowly added, warmed to room temperature and stirred for 0.5 hours. The above reaction solution was cooled to 0 ° C under nitrogen protection, methanol (10 mL) was slowly added dropwise, and the residue obtained after the reaction solution was concentrated was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2: 1) to obtain compound 6 (2.0 g, yield 100%) as a white solid.

[0282] LCMS: [M+H] + =270.0.

[0283] Step 4: Compound 8

[0284] Compound 6 (2.0 g, 7.0 mmol, 1.0 eq), compound 7 (1.65 g, 14.8 mmol, 2.1 eq), potassium iodide (1.23 g, 7.0 mmol, 1.0 eq) and potassium carbonate (2.35 g, 22 mmol, 3.0 eq) were added to dimethylformamide (10 mL) under nitrogen protection, and the reaction solution was heated to 90 ° C and stirred for 16 hours. After cooling to room temperature, the reaction solution was diluted with ethyl acetate (200 mL) and filtered, the filtrate was washed with saturated sodium chloride solution, the organic phase was dried over anhydrous sodium sulfate, and the residue obtained after concentration was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2: 1) to obtain compound 8 (1.0 g, yield 45%) as a white solid.

[0285] LCMS: [M+H] + =301.1

[0286] Step 5: Compound 10

[0287] Compound 8 (600 mg, 2.0 mmol, 1.0 eq) was dissolved in dichloromethane (20 mL), and compound 9 (296 mg, 1.0 mmol, 0.5 eq) was added, and stirred at room temperature for 1 hour under nitrogen protection. The reaction solution was cooled to 0 ° C, aluminum chloride (800 mg, 6.0 mmol, 3.0 eq) was added, and the temperature was slowly raised to room temperature and stirred for 16 hours. The reaction solution was diluted with dichloromethane (100 mL), washed with ice water, and the organic phase was dried over anhydrous sodium sulfate. The residue obtained after concentration was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1) to obtain compound 10 (200 mg, yield 30%) as a white solid.

[0288] LCMS: [M+H] + =327.1

[0289] Step 6: Compound 11

[0290] Compound 10 (190 mg, 0.6 mmol) was dissolved in concentrated hydrochloric acid (12 M, 15 mL), heated to 99°C and stirred for 64 hours under nitrogen protection. After cooling to room temperature, the reaction solution was concentrated, and the residue was azeotropically dehydrated with toluene and ethanol to obtain crude compound 11 (200 mg) as a white solid.

[0291] LCMS: [M+H] + =285.2

[0292] Step 7: Compound 12

[0293] Compound 11 (200 mg, 0.6 mmol) was dissolved in tetrahydrofuran (50 mL), cooled to 0°C, and LiAlD4 (252 mg, 6.0 mmol, 10.0 eq) was slowly added, and then the temperature was raised to room temperature and stirred for 16 hours. The reaction solution was diluted with tetrahydrofuran (250 mL) and cooled to 0°C, and sodium sulfate decahydrate (10 g) was added and stirred for 0.5 hours. The reaction solution was filtered and concentrated, and the residue obtained was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1) to obtain compound 12 (25 mg, two-step yield 15%) as a yellow solid.

[0294] LCMS: [M+H] + =273.1

[0295] Step 8: Compound M

[0296] Compound 13 (21 mg, 0.183 mmol, 2.0 eq), HATU (70 mg, 0.183 mmol, 2.0 eq) and diisopropylethylamine (36 mg, 0.275 mmol, 3.0 eq) were dissolved in dimethylformamide (5 mL) and stirred at room temperature for 0.5 hours under nitrogen protection. Compound 12 (25 mg, 0.092 mmol, 1.0 eq) was added to the above reaction solution, and the temperature was raised to 45 ° C and stirred for 16 hours. The reaction solution was diluted with ethyl acetate (50 mL), washed with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate. The residue obtained after concentration was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1) to obtain compound M (2.3 mg, yield 6.8%) as a white solid.

[0297] LCMS: [M+H] + =369.2

[0298] 1 H NMR (400MHz, CDCl3) δ7.18 (s, 1H), 7.01-6.86 (m, 5H), 3.47 (t, J = 5.6Hz, 2H), 2.81 (t, J=5.6Hz,2H),2.38(s,2H),2.22(s,3H),2.12(s,3H),1.28(s,3H),0.60-0.50(m,4H).

[0299] Example 14 Preparation of Compound N

[0300]

[0301] Step 1: Compound N

[0302] Compound 1 (30 mg, 0.11 mmol, 1.0 eq) was dissolved in toluene (13 mL), and cesium carbonate (384 mg, 0.33 mmol, 3.0 eq), compound 2 (37 mg, 0.17 mmol, 1.5 eq), x-Phos (11 mg, 0.02 mmol, 0.2 eq) and Pd2(dba)3 (11 mg, 0.01 mmol, 0.1 eq) were added in sequence, and the mixture was heated to 90 ° C and stirred for 15 hours under nitrogen protection. The reaction solution was cooled to room temperature and diluted with water, extracted with ethyl acetate (3×20 mL), and the combined organic phase was washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The concentrated residue was purified by Pre-HPLC (0.1% formic acid / acetonitrile / water) to obtain compound N (2 mg, yield 5%) as a white solid.

[0303] LCMS: [M+H] + =363.2

[0304] 1 H NMR(400MHz,DMSO-d6)δ9.02(s,1H),7.04-7.02(m,2H),6.93-6.91(m,3H),4.25(s,2H),3.49(s,2H),2.51-2.49 (m,2H),2.21(s,2H),2.20(s,3H),2.11(s,3H),2.01(s,3H),1.15(s,3H),0.56-0.53(m,2H),0.33-0.31(m,2H).

[0305] Example 15 Preparation of Compound O

[0306]

[0307] Step 1: Compound O

[0308] Compound 1 (50 mg, 0.183 mmol, 1.0 eq) was dissolved in dimethyl sulfoxide (5 mL), and compound 2 (33 mg, 0.275 mmol, 1.5 eq) and potassium carbonate (76 mg, 0.275 mmol, 1.5 eq) were added, and the mixture was heated to 100°C and stirred for 16 hours. After cooling to room temperature, the mixture was diluted with water and extracted with ethyl acetate (3 x 20 mL). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The residue obtained after concentration was purified by pre-HPLC (0.1% ammonia water / acetonitrile / water) to obtain compound O (4.2 mg, yield 6%) as a white solid.

[0309] LCMS: [M+H] + =374.1

[0310] 1 H NMR (400MHz, DMSO-d6) δ9.02(s,1H),7.56(d,J=9.0Hz,2H),7.04(d,J=9.1Hz,2H),6.92(s,1H),4.47(s,2H),3.66(s,2H) ,2.74(t,J=5.7Hz,2H),2.18(s,2H),2.09(s,3H),2.00(s,3H),1.12(s,3H),0.51(q,J=4.2Hz,2H),0.29(q,J=4.1Hz,2H).

[0311] Example 16 Preparation of Compound P

[0312]

[0313] Step 1: Compound P

[0314] Compound 1 (30 mg, 0.110 mmol, 1.0 eq) was dissolved in toluene (5 mL), and compound 2 (45 mg, 0.165 mmol, 1.5 eq), potassium tert-butoxide (37 mg, 0.330 mmol, 3.0 eq), X-Phos (11 mg, 0.022 mmol, 0.2 eq) and Pd2(dba)3 (10 mg, 0.011 mmol, 0.1 eq) were added, and the mixture was heated to 100°C and stirred for 16 hours under nitrogen protection. After cooling to room temperature, the reaction solution was diluted with ethyl acetate (30 mL), and then washed with water and saturated sodium chloride solution in sequence. The organic phase was dried over anhydrous sodium sulfate, and the residue obtained after concentration was purified by Pre-HPLC (0.1% formic acid / acetonitrile / water) to obtain compound P (6.3 mg, yield 13%) as a white solid.

[0315] LCMS: [M+H] + =417.1

[0316] 1 H NMR(400MHz, DMSO-d6)δ9.05(s,1H),7.51(d,J=8.7Hz,2H),7.12(d,J=8.7Hz,2H),6.96(s,1H),4.46(s,2H),3 .68(s,2H),2.77(s,2H),2.22(s,2H),2.12(s,3H),2.03(s,3H),1.15(s,3H),0.55(s,2H),0.33-0.31(m,2H).

[0317] Example 17 Preparation of Compound Q

[0318]

[0319] Step 1: Compound Q

[0320] Compound 1 (40 mg, 0.15 mmol, 1.0 eq) and compound 2 (51.6 mg, 0.22 mmol, 1.5 eq) were dissolved in toluene (5 mL), Pd2(dba)3 (14 mg, 0.015 mmol, 0.1 eq), Xant-Phos (14 mg, 0.03 mmol, 0.2 eq) and cesium carbonate (143 mg, 0.45 mmol, 3.0 eq) were added, and the temperature was raised to 90°C under nitrogen protection for 16 hours. After cooling to room temperature, the reaction solution was concentrated, and the obtained residue was purified by Pre-HPLC (0.1% formic acid / acetonitrile / water) to obtain compound Q (9.7 mg, yield 18%) as a white solid.

[0321] LCMS: [M+H] + =379.2

[0322] 1 H NMR(400MHz, CDCl3)δ7.18(s,1H),6.98(d,J=8.8Hz,2H),6.92–6.83(m,3H),4.23(s,2H),3.78(s,3H),3.44(t ,J=5.6Hz,2H),2.83(t,J=5.6Hz,2H),2.40(s,2H),2.23(s,3H),2.13(s,3H),1.30(s,3H),0.61–0.54(m,4H).

[0323] Example 18 Preparation of Compound R

[0324]

[0325] Step 1: Compound R

[0326] Compound 1 (30 mg, 0.11 mmol, 1.0 eq), compound 2 (33 mg, 0.17 mmol, 1.5 eq), sodium tert-butoxide (32 mg, 0.33 mmol, 3.0 eq), Dave-Phos (17 mg, 0.044 mmol, 0.4 eq) and Pd2(dba)3 (20 mg, 0.022 mmol, 0.2 eq) were dissolved in 1,4-dioxane (4 mL) and tert-butanol (2 mL), and the temperature was raised to 99°C under nitrogen protection for 18 hours. After cooling to room temperature, the reaction solution was extracted with ethyl acetate and saturated sodium chloride solution, the organic phase was dried over anhydrous sodium sulfate, and the residue obtained after concentration was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain compound R (26 mg, yield 60%) as a yellow solid

[0327] LCMS: [M+H] + =392.2

[0328] 1 H NMR(400MHz,MeOD)δ7.15(s,1H),7.01(d,J=7.6Hz,2H),6.93(d,J=8.0Hz,2H),4.50(s,2H),2.93-2.90(m,2H),2.85–2.83( m,2H),2.81(s,3H),2.80(s,3H),2.23(s,2H),2.11(s,3H),2.02(s,3H),1.12(s,3H),0.53-0.50(m,2H),0.33-0.31(m,2H).

[0329] Example 19 Preparation of Compound S

[0330]

[0331] Step 1: Compound 3

[0332] Compound 1 (50 mg, 0.18 mmol, 1.0 eq) was dissolved in tetrahydrofuran (15 mL), and compound 2 (83 mg, 0.26 mmol, 1.5 eq), Pd2(dba)3 (17 mg, 0.018 mmol, 0.1 eq), Dave-Phos (14 mg, 0.036 mmol, 0.2 eq) and LiHMDS (1 M, 1.8 mL, 1.8 mmol, 10 eq) were added, and the temperature was raised to 80 ° C. and stirred for 4 hours under nitrogen protection. After the reaction solution was cooled to room temperature, it was extracted with ethyl acetate and saturated sodium chloride solution, the organic phase was dried over anhydrous sodium sulfate, and the residue obtained after concentration was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to obtain compound 3 (4 mg, yield 5%) as a yellow solid.

[0333] LCMS: [M+H] + =445.2

[0334] Step 2: Compound S

[0335] Compound 3 (4 mg, 0.009 mmol, 1.0 eq) and potassium carbonate (4 mg, 0.027 mmol, 3.0 eq) were added to methanol (5 mL) and stirred at room temperature for 2 hours. The reaction solution was filtered and the obtained solution was concentrated to obtain compound S (3 mg, yield 90%) as a yellow solid.

[0336] LCMS: [M+H] + =373.2

[0337] Example 20 Preparation of Compound T

[0338]

[0339] Step 1: Compound 3

[0340] Compound 1 (80 mg, 0.293 mmol, 1.0 eq) was dissolved in toluene (5 mL), and compound 2 (129 mg, 0.587 mmol, 2.0 eq), sodium tert-butoxide (56 mg, 0.587 mmol, 2.0 eq), BINAP (37 mg, 0.0587 mmol, 0.2 eq) and Pd2(dba)3 (27 mg, 0.0293 mmol, 0.1 eq) were added, and the mixture was heated to 100°C and stirred for 16 hours under nitrogen protection. After cooling to room temperature, the reaction solution was diluted with ethyl acetate (30 mL), and then washed with water and saturated sodium chloride solution in turn, the organic phase was dried over anhydrous sodium sulfate, and the residue after concentration was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1) to obtain compound 3 (30 mg, yield 25%) as a brown solid.

[0341] LCMS: [M+H]+=413.1

[0342] Step 2: Compound 4

[0343] Compound 3 (20 mg, 0.0484 mmol, 1.0 eq) was dissolved in tetrahydrofuran (3 mL), lithium aluminum tetrahydride (9 mg, 0.2424 mmol, 5.0 eq) was added, and stirred at room temperature for 10 minutes under nitrogen protection, tetrahydrofuran (5 mL) was added to dilute and the reaction solution was cooled to 0°C, sodium sulfate decahydrate (0.3 g) was added and stirred at 0°C for 15 minutes. The reaction solution was filtered and concentrated to obtain compound 4 (15 mg, yield 80%) as a brown solid.

[0344] LCMS: [M+H]+=385.1

[0345] Step 2: Compound T

[0346] Compound 4 (10 mg, 0.026 mmol, 1.0 eq) was dissolved in anhydrous ethanol (1 mL), triethylsilane (3 mg, 0.026 mmol, 1.0 eq) and palladium chloride (2 mg, 0.013 mmol, 0.5 eq) were added, and stirred at room temperature for 1 hour under nitrogen protection. The reaction solution was filtered and concentrated to obtain compound T (8 mg, 83%) as a brown solid.

[0347] LCMS: [M+H]+=369.1

[0348] Example 21 Preparation of Compound U

[0349]

[0350] Step 1: Compound U

[0351] Compound 1 (120 mg, 0.4 mmol, 1.0 eq) was dissolved in toluene (40 mL), and compound 2 (143 mg, 0.6 mmol, 1.5 eq), cesium carbonate (428.1 mg, 1.31 mmol, 3.0 eq), X-Phos (41.7 mg, 0.08 mmol, 0.2 eq) and Pd2(dba)3 (40.1 mg, 0.04 mmol, 0.1 eq) were added, and the mixture was heated to 90°C and stirred for 16 hours under nitrogen protection. After cooling to room temperature, the reaction solution was filtered, the filtrate was diluted with water (30 mL), extracted with ethyl acetate (3 x 50 mL), the combined organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the residue obtained after concentration was purified by neutral alumina silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to obtain compound U (42.3 mg, yield 26%) as a white solid.

[0352] LCMS: [M+H] + =365.2

[0353] 1 H NMR (400MHz, DMSO) δ9.09 (s, 1H), 7.03 (d, J = 8.0Hz, 2H), 6.93-6.90 (m, 3H), 4.24 (s, 2H), 3.50-3 .47(m,2H),2.72-2.66(m,2H),2.21(s,2H),2.19(s,3H),2.11(s,3H),2.00(s,3H),1.06(s,9H).

[0354] Example 22 Potassium Channel Opener Activity Test (FDSS / μCELL Detection)

[0355] 1. Experimental methods:

[0356] 1.1 Experimental Procedure

[0357] Cell preparation: CHO-KCNQ2 cells were cultured in 175 cm 2 In the culture flask, when the cell density grows to 60-80%, remove the culture medium, wash once with 7mL PBS (Phosphate Buffered Saline), and then add 3mL 0.25% Trypsin for digestion. After digestion is complete, add 7mL culture medium (90% DMEM / F12+10% FBS+500μg / mL G418) for neutralization, centrifuge at 800rpm for 3 minutes, aspirate the supernatant, add 5mL culture medium to resuspend, and count the cells.

[0358] Cell plating: According to the cell counting results, adjust the density to 3x10 4 After standing at room temperature for 30 minutes, the cells were placed in a 37°C CO2 incubator for overnight culture. After 16-18 hours of culture, the cell density reached about 80%.

[0359] Fluorescent dye incubation: discard the cell culture medium, add 80 μL / well loading buffer, and incubate at room temperature in the dark for 60 minutes.

[0360] Compound incubation: discard the loading buffer, add 80 μL / well of the prepared compound solution, and incubate at room temperature in the dark for 20 minutes.

[0361] Fluorescence data acquisition: Real-time fluorescence signal recording was performed using the FDSS / μCELL instrument, with an excitation wavelength of 480 nm and an emission wavelength of 540 nm. The signal was recorded once per second. After recording a 10-second baseline, 20 μL / well of stimulation buffer was added, and the recording was continued until 180 seconds.

[0362] 1.2 Solution preparation

[0363] Loading buffer: 10 mL / plate, prepared as follows:

[0364] Element volume <![CDATA[PowerLoad TM Concentrate, 100X (ingredient C)]]> 100μL <![CDATA[FluxOR TM Reagents, reconstituted in DMSO (step 1.2)]]> 10μL Deionized water 8.8mL <![CDATA[FluxOR TM Assay Buffer, 10X (Component B)]]> 1mL Probenecid, reconstituted in deionized water (step 1.1) 100μL Total volume 10mL

[0365] Test buffer sample: 100mL / plate, prepared as follows:

[0366] Element volume Deionized water 8.9mL <![CDATA[FluxOR TM Assay Buffer, 10X (Component B)]]> 1mL Probenecid, reconstituted in deionized water (step 1.1) 100μL Total volume 10mL

[0367] Stimulation buffer: 5 mL / plate, prepared as follows:

[0368]

[0369] The above buffer is from a commercially available kit named FluxOR potassium ion channel assay.

[0370] 1.3 Compound preparation

[0371] Prepare 20mM DMSO compound stock solution, take 10μL of 20mM compound stock solution to 20μL DMSO solution, and dilute 3-fold to 8 intermediate concentrations; then take the intermediate concentration compounds to the test buffer, dilute 200-fold to get the final concentration to be tested, and take 80μL to add to the test plate.

[0372] The highest tested concentration was 100 μM, followed by 8 concentrations, namely 100, 33.33, 11.11, 3.70, 1.23, 0.41, 0.137, and 0.045 μM, with 3 replicates for each concentration.

[0373] The DMSO content in the final test concentration did not exceed 0.5%, which is a concentration that has no effect on KCNQ2 potassium channels.

[0374] 1.4 Data Analysis

[0375] The experimental data were analyzed by Excel 2007 and GraphPad Prism 5.0 software, and the ratio of 180 seconds was statistically calculated to calculate the agonistic effect. The agonistic effect of the compound was calculated by the following formula:

[0376]

[0377] 1.5 Quality Control

[0378] Environment: Temperature ~25℃

[0379] Reagents: FluxORTM Detection Kit (Invitrogen, Cat#F0017)

[0380] The experimental data in the report must meet the following criteria: Z'Factor>0.5

[0381] 2. Determination results: See Table 1 for details, where EC 50 The smaller the value, the higher the activity of the corresponding compound.

[0382] Table 1. Test results of some compounds described in the present invention

[0383] Compound Maximum excitation rate (%) <![CDATA[EC 50 (uM)]]> A 37.97 0.44 B 37.5 0.16 D 29.51 0.45 I 38.3 0.44 U 52.3 0.33

[0384] References for the above test methods:

[0385] Zhaobing Gao et al.Journal of Biological Chemistry.2010,285(36):28322-28332.

[0386] Example 23 Pharmacokinetic Study in Mice

[0387] 1) Research purpose: To obtain the pharmacokinetic characteristics and blood-brain barrier status of the test compound in male ICR mice

[0388] 2) Experimental content

[0389] Six healthy male ICR mice (weight range 18-22 g) were taken and divided into 2 groups, 3 mice / group. After fasting overnight (oral administration group only), the test compound was administered intravenously at 0.5 mg / kg and orally at 10 mg / kg. Blood was collected by jugular vein puncture at time points 0.083 (IV only), 0.25, 0.5, 1, 2, 4, 6 (PO only), 8 and 24 h. At least 0.3 mL of whole blood was collected into EDTA-K2 anticoagulant tubes. Within half an hour, plasma was obtained by centrifugation (6000 rpm, 8 minutes, 4°C) and frozen at -20°C for later use.

[0390] Brain-to-blood ratio study: 6 healthy male ICR mice (weight range 18-22 g) were divided into 2 groups, 3 mice / group, and the test compound (10 mg / kg) was orally administered after overnight fasting. Blood was collected by cardiac puncture at time points 2 and 4 h, and at least 0.5 mL of whole blood was collected into EDTA-K2 anticoagulant tubes. Within half an hour, plasma was centrifuged (6000 rpm, 8 minutes, 4°C) and frozen at -20°C for later use. At the same time, brain tissue was collected, rinsed with physiological saline, dried with absorbent paper, weighed, and frozen at -20°C for later use.

[0391] Experimental results: According to the obtained blood drug concentration data, The pharmacokinetic parameters after drug administration were calculated using the non-compartmental model using Pharsight 7.0 software (Pharsight, USA).

[0392] Table 2 PK parameters of single administration in male ICR mice

[0393]

[0394]

[0395] * The intravenous (IV) dose is 1 mg / kg

[0396] Table 3 Brain-to-blood ratio at different time points after single oral administration in male ICR mice

[0397]

[0398] The brain-to-blood ratio is crucial for neurological drugs. As shown in Table 3, the compounds of the present invention all have excellent brain-to-blood ratios, which will bring better efficacy and excellent safety.

[0399] Example 24 Pharmacokinetic Study in Rats

[0400] 1) Research purpose: To obtain the pharmacokinetic characteristics of the test compound in male SD rats.

[0401] 2) Experimental content

[0402] Pharmacokinetic study: Six healthy male SD rats (SPF grade) were divided into two groups, 3 rats / group, and compound A was administered intravenously at 0.3 mg / kg and orally at 5 mg / kg after overnight fasting (oral administration group only). Blood was collected by jugular vein puncture at time points of 0.083 (IV only), 0.25, 0.5, 1, 2, 4, 6 (PO only), 8 and 24 h. At least 0.3 mL of whole blood was collected into EDTA-K2 anticoagulant tubes. Within half an hour, plasma was obtained by centrifugation (6000 rpm, 8 minutes, 4°C) and frozen at -20°C for later use.

[0403] Experimental results: According to the obtained blood drug concentration data, The pharmacokinetic parameters after drug administration were calculated using the non-compartmental model using Pharsight 7.0 software (Pharsight, USA).

[0404] Table 4 PK parameters of single administration in male SD rats

[0405]

[0406] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A compound represented by formula A or a pharmaceutically acceptable salt thereof, in, R1 is substituted or unsubstituted phenyl, wherein the substitution refers to substitution by one or more substituents selected from the group consisting of halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, -NR8R9, ethynyl; R2 and R3 are independently selected from the following group: hydrogen, deuterium, C 1-6 Alkyl, halogenated C 1-6 alkyl; R4 and R5 are independently selected from the following groups: hydrogen, C 1-6 Alkyl, halogenated C 1-6 alkyl; R6 is selected from substituted or unsubstituted C3 cycloalkyl, wherein the substitution refers to substitution by one or more substituents selected from the following group: 1-6 Alkyl, halogenated C 1-6 alkyl; n is 1; W is C-R7; R7 is selected from the group consisting of hydrogen, deuterium, C 1-6 Alkyl, halogenated C 1-6 alkyl; R8 and R9 are independently selected from the following groups: hydrogen, C 1-6 Alkyl, halogenated C 1-6 alkyl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that R1 is substituted or unsubstituted phenyl, wherein the substitution refers to substitution by one or more substituents selected from the group consisting of halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, -NR8R9, ethynyl; R2 and R3 are independently selected from the group consisting of hydrogen, deuterium; R4 and R5 are methyl groups; R6 is selected from substituted or unsubstituted C3 cycloalkyl, wherein the substitution refers to substitution by one or more substituents selected from the following group: 1-6 alkyl; n is 1; W is C-R7; R7 is hydrogen; R8 and R9 are independently C 1-6 alkyl.

3. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, characterized in that: R1 is substituted or unsubstituted phenyl, wherein the substitution refers to substitution by one or more substituents selected from the group consisting of halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, -NR8R9, ethynyl; R2 and R3 are independently selected from the group consisting of hydrogen, deuterium; R4 and R5 are methyl groups; R6 is selected from the group consisting of: n is 1; W is C-R7; R7 is hydrogen; R8 and R9 are independently C 1-6 alkyl.

4. The compound according to claim 3 or a pharmaceutically acceptable salt thereof, characterized in that: R1 is substituted or unsubstituted phenyl, wherein the substitution refers to substitution by one or more substituents selected from the group consisting of halogen, C 1-6 Alkyl, halogenated C 1-6 alkyl; R2 and R3 are independently selected from the group consisting of hydrogen, deuterium; R4 and R5 are methyl groups; R6 is n is 1; W is C-R7; R7 is hydrogen; R8 and R9 are independently C 1-6 alkyl.

5. A method for preparing the compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: Includes steps: in: X is selected from the group consisting of halogen, -B(OH)2, -OTf; R1, R2, R3, R4, R5, R6, n, and W are as defined in claim 1.

6. A pharmaceutical composition, characterized in that It comprises one or more pharmaceutically acceptable carriers and a therapeutically effective amount of one or more compounds according to claim 1 or pharmaceutically acceptable salts thereof.

7. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: Used for preparing drugs for preventing and / or treating diseases sensitive to potassium ion channels.

8. The use according to claim 7, characterized in that The disease sensitive to potassium ion channels is a central nervous system disease.

9. The use according to claim 8, characterized in that The central nervous system disease is selected from the group consisting of epilepsy, convulsions, inflammatory pain, neuropathic pain, migraine, depression, anxiety disorders, stroke, neurodegenerative diseases, cocaine abuse, nicotine withdrawal, alcohol withdrawal, and tinnitus.

10. The use according to claim 9, characterized in that The neurodegenerative disease is Alzheimer's disease.

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