CN116789644A - Amide compound and preparation method and application thereof - Google Patents

Abstract

The invention provides an amide compound as shown in a formula I, and a preparation method and application thereof. The amide compound has Nav1.8 selective inhibition activity, can be used as a Nav1.8 selective inhibitor, has better activity, higher selectivity and fewer side effects, can be used for treating, preventing or controlling diseases related to Nav1.8 channel participation or dysfunction, and has important clinical application value.

Description

Amide compound and preparation method and application thereof

Technical Field

The invention relates to the technical field of inhibitor synthesis, in particular to an amide compound, a preparation method thereof and application thereof in treating Nav1.8 target pain related diseases.

Background

Pain acts as a protective mechanism that alerts and protects the tissue from further injury. Pain is produced primarily by the transformation of stimulus received by nociceptors into nerve impulses (action potentials) and transmitted to the nerve center via afferent nerve fibers, causing pain sensation, whereas action potential generation and conduction in neurons depends on voltage-gated sodium ion channels on the cell membrane (voltage-gated sodium channels, nav).

Voltage-gated sodium ion channels mediate sodium ion selective transmembrane flow and play a key role in initiating, conducting, and delivering action potentials in excitable cells such as neurons and the like (Catterall et al, pharmacol Rev.2005,57 (4): 397-409.). Nav channels are important drug targets and Nav channel inhibitors are used in the treatment of pain, arrhythmias, epilepsy, anaesthesia, itch and other diseases (Black et al, neuron.2013,80 (2): 280-91; catterall et al, annu Rev Pharmacol Toxicol.2014,54:317-38; bennett et al, physiol Rev.2019,99 (2): 1079-1151.). Currently, 9 channel subtypes Nav1.1 through Nav1.9 channels have been found in the mammalian genome. According to the homology of amino acid sequences, the similarity of Nav channel proteins is between 45% and 87%. TTX-resistant channels (TTX-R) and TTX-sensitive channels (TTX-S) are classified according to their sensitivity To Tetrodotoxin (TTX), with Nav1.5, nav1.8 and Nav1.9 channels belonging to the TTX-R sodium channel and other subtypes belonging to the TTX-S subtype (Catterall et al, pharmacol Rev.2005,57 (4): 397-409.).

The voltage-gated Nav1.8 channel subtype (TTX-R type) is predominantly distributed in the peripheral nervous system, e.g., 75% of dorsal root neurons express Nav1.8 channels. Because of the relatively high activation and deactivation voltages of the Nav1.8 channel, it becomes the major component of the action potential rising branch (other Nav channel subtypes are already in a nonfunctional inactive state) (Goodwin et al, nat Rev Neurosci.2021,22 (5): 263-274). Due to the slow inactivation and fast reactivation characteristics of the Nav1.8 channel, it is involved in the physiological and pathological processes of membrane potential depolarization and neuronal high frequency discharge, such as pain (alloum et al, nat Rev neuron.2020, 16 (12): 689-705). Human genetics studies have shown that mutations in the Nav1.8 gene lead to small fiber neuralgia and erythema pain (Faber et al, proc Natl Acad Sci U S A.2012,109 (47): 19444-9; kaluza et al, pflugers arch.2018,470 (12): 1787-1801.). In rodents, gene knockout or knockout of the nav1.8 channel gene can alleviate a variety of inflammatory and neuropathic pain; whereas administration of Nav1.8 channel inhibitors such as A-803467 is effective in alleviating pain response (Jarvis et al Proc Natl Acad Sci U S A.2007,104 (20): 8520-5.). Diabetic neuralgia is one of the most common neuropathic pain disorders, about 60% to 70% of diabetics suffer from this disease, and more than 70% of patients are not effectively treated (Jensen et al, brain.2021,144 (6): 1632-1645.). Methylglyoxal in diabetic neuralgia patients directly enhances the function of the nav1.8 channel, and gene knockout or knockout of the nav1.8 channel is effective in alleviating neuralgia (Bierhaus et al, nat med.2012,18 (6): 926-33.). In the STZ-induced diabetic neuralgia rat model, the administration of Nav1.8 channel inhibitor A-803467 to the abdominal cavity or plantar surface can provide dose-dependent relief of pain behavioural responses in animals (Mert et al J Am Assoc Lab Anim Sci.2012,51 (5): 579-85.).

In addition to pain, the Nav1.8 channel is also associated with multiple sclerosis, arrhythmia, cough, itch, and epilepsy. Multiple sclerosis (Multiple sclerosis, MS) is an inflammatory demyelinating disease that is primary in the central nervous system, the exact pathogenesis of which has yet to be elucidated. The normal human cerebellum purkinje fiber does not express Nav1.8 channels, the expression of the cerebellum Nav1.8 in patients with multiple sclerosis is up-regulated, and the expression level of the channels appears to increase dependently with the development of the course of the disease, and the Single Nucleotide Polymorphism (SNP) of the Nav1.8 coding gene is also related to the incidence of MS (Craner et al, J Neuropathol Exp neurol.2003,62 (9): 968-75; rootaei et al, neurology.2016,86 (5): 410-7.). Mice with Nav1.8 (overexpressed) knocked-in Nav1.8 fibers (L7-1.8 TG) exhibited multiple sclerosis behavior, and administration of Nav1.8 selective inhibitor PF-01247324 alleviated MS behavior in L7-1.8TG transgenic mice (Shields et al, ann neurol 2012,71 (2): 186-94; shields et al, PLoS one.2015,10 (3): e 0119067.). Osteoarthritis is a degenerative osteoarthropathy, the major features of which are cartilage wear and pain. Phosphorylated cAMP response element binding protein (CREB) binds directly to the promoter of the Nav1.8 encoding gene, promoting transcription of the Nav1.8 protein, up-regulating Nav1.8 channel expression levels (Zhu et al, elife.2020, 9:e57656.). In the cardiovascular system, nav1.8 channels have been demonstrated to be expressed in cardiac nerves such as Purkinje fibers, and some studies have suggested that Nav1.8 is also expressed in cardiac myocytes (Verkerk et al, circ Res.2012,111 (3): 333-43.). Human genetics studies have found that Nav1.8 gene mutations are associated with Brugada syndrome (Hu et al, J Am Coll cardiol.2014,64 (1): 66-79.). Inhibition of the Nav1.8 channel improves heart remodeling, and the Nav1.8 channel is considered as a potential therapeutic target for cardiovascular diseases such as arrhythmia, atrial fibrillation, heart failure and the like (Dybkova et al, cardiovasc Res.2018,114 (13): 1728-1737.). Nav1.8 channels are expressed in the vagal plexus associated with cough, and Nav1.8 phosphorylation levels and expression levels are elevated during pathological cough, involving in cough reflex (Muroi et al, lung.2014,192 (1): 15-20.). In the itch experience of mammals, itch factors such as histamine released by lymphocytes, mast cells and the like can activate Nav1.8 channels, and knockout of Nav1.8 of mice can effectively relieve itching behaviors induced by histamine and endothelin (Riol-Blanco et al, 2014,510 (7503):157-61.). In addition, congenital mutations in human Nav1.8 have been reported to cause epilepsy, convulsive disorders (Kambouris et al, ann Clin Transl Neurol.2016,4 (1): 26-35.).

Currently, a Nav1.8 selective inhibitor is VX-150 from VERTEX, which has achieved positive results in stage II clinical in patients with osteoarthritis, acute pain and pain caused by small fiber neuropathy. The Nav1.8 selective inhibitor entering the clinic in China is Hengrui HRS-4800, and a phase I clinical experiment is currently being carried out; other multiple selective inhibitors are in preclinical development stages; therefore, the Nav1.8 inhibitor with better development activity, higher selectivity and fewer side effects has important clinical application and innovative pharmaceutical value.

In view of this, the present invention has been made.

Disclosure of Invention

An object of the present invention is to provide an amide compound having nav1.8 selective inhibitory activity.

The second object of the present invention is to provide a method for producing an amide compound.

The invention also aims to provide a pharmaceutical composition containing the amide compound.

The fourth object of the invention is to provide an application of the amide compound or the pharmaceutical composition in preparing Nav1.8 inhibitor or preparing medicines for treating, preventing or controlling diseases or symptoms related to Nav1.8 channels.

In order to achieve the above object of the present invention, the following technical solutions are specifically adopted:

in one aspect, the present invention provides a compound of formula I, an isomer, a racemate, a prodrug or a pharmaceutically acceptable salt thereof,

wherein:

x is selected from N or CH; y is selected from N or CR ₃ ；

V and G are each independently selected from N or CH, Q and T are each independently selected from N or C;

A. w and Z are each independently selected from O, S, N, carbonyl, sulfoxide, sulfone, -NR _a -、-CR _b -、-NR _a -CO-、-CR _b ＝N-、-CR _b -NR _a -, and at least one of A, W and Z contains nitrogen;

R _a independently at each occurrence selected from hydrogen, amino, hydroxy, C1-C6 alkyl, halo C1-C6 alkyl, C1-C6 alkoxy, halo C1-C6 alkoxy, C1-C6 alkylamino, C3-C8 cycloalkyl, 3-8 membered heterocyclyl containing 1 to 4 heteroatoms selected from N, O, S, C6-C12 aryl, or 5-10 membered heteroaryl containing 1 to 4 heteroatoms selected from N, O, S;

R _b independently at each occurrence selected from hydrogen, halogen, nitro, amino, cyano, hydroxy, C1-C6 alkyl, halo C1-C6 alkyl, C1-C6 alkoxy, halo C1-C6 alkoxy, C1-C6 alkylamino, C3-C8 cycloalkyl, 3-8 membered heterocyclyl containing 1 to 4 heteroatoms selected from N, O, S, C6-C12 aryl, or 5-10 membered heteroaryl containing 1 to 4 heteroatoms selected from N, O, S;

n is selected from 0, 1 or 2; in particular 2;

R ₁ selected from hydrogen, hydroxy, halogen, cyano, nitro or amino; preferably hydrogen, fluorine, chlorine, bromine, amino or hydroxyl;

R ₂ selected from halogen, hydroxy, cyano, nitro or halogenated C1-C6 alkyl; preferably chlorine, bromine, iodine, trifluoromethyl;

R ₃ selected from halogen, hydroxy, cyano, amino, C1-C6 alkyl, halo C1-C6 alkyl, C2-C6 alkenyl, halo C2-C6 alkenyl, C2-C6 alkenyloxy, halo C2-C6 alkenyloxy, C2-C6 alkynyl, halo C1-C6 alkynyl, C2-C6 alkynyloxy, halo C1-C6 alkynyloxy, C1-C6 alkoxy, halo C1-C6 alkoxy, C1-C6 alkylamino, halo C1-C6 alkylamino, C3-C6 cycloalkylamino, C1-C6 alkylamino, C3-C6 cycloalkyl, C3-C6 cycloalkoxy or a 3-8 membered heterocyclyl containing 1 to 4 heteroatoms selected from N, O, S; preferably fluorine, chlorine, bromine, amino, hydroxyl, methyl, cyclopropyl, methoxy, trifluoromethyl, trifluoromethoxy;

R ₆ and R is ₇ Each independently selected from hydrogen, fluorine, chlorine, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy; preferably each independently is hydrogen, fluorine, chlorine, methyl, ethyl or isopropyl;

R ₅ 、R ₈ and R is ₉ Each independently selected from hydrogen, fluorine, chlorine, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl; preferably each independently is hydrogen, methyl, ethyl, isopropyl or cyclopropyl;

Represents a single bond or a double bond.

In some embodiments, the compound of formula I is selected from the following compounds of formula II:

wherein R is ₃ The definitions of A, Z, W, Q, T, V and G are the same as previously described.

In some preferred embodiments of the present invention,is one selected from the following groups:

in some preferred embodiments, the compound of formula I is selected from the following compounds:

the terms in the present invention are defined as follows:

"halogen" may be fluorine, chlorine, bromine or iodine.

"C1-C6" alkyl refers to a chain alkyl group having 1 to 6 carbon atoms; specific examples thereof may include methyl, ethyl, propyl, n-propyl, isopropyl, butyl, n-butyl, isobutyl, t-butyl, 1-methyl-butyl, 1-ethyl-butyl, pentyl, n-pentyl, isopentyl, neopentyl, t-pentyl, hexyl, n-hexyl, 1-methylpentyl, 2-methylpentyl, 4-methyl-2-pentyl, 3-dimethylbutyl, 2-ethylbutyl and the like; "haloalkyl" refers to a group as described above wherein at least one hydrogen of the alkyl is replaced with a halogen; specific examples thereof include trifluoromethyl and the like.

"C2-C6 alkenyl" refers to a straight or branched chain group containing 2 to 6 carbon atoms and having at least one carbon-carbon double bond; specific examples thereof may include ethenyl, propenyl, 2-propenyl, (E) -2-butenyl, (Z) -2-butenyl, (E) -2-methyl-2-butenyl, (Z) -2-methyl-2-butenyl, 2, 3-dimethyl-2-butenyl, (Z) -2-pentenyl, (E) -1-pentenyl, (E) -2-pentenyl, (Z) -2-hexenyl, (E) -1-hexenyl, (Z) -1-hexenyl, (E) -2-hexenyl, (Z) -3-hexenyl, (E) -1, 3-hexadienyl, 4-methyl-3-pentenyl or norbornene.

"C2-C6 alkynyl" refers to a straight or branched chain group containing 2 to 6 carbon atoms and having at least one carbon-carbon double bond; specific examples thereof may include ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl.

"C1-C6 alkoxy" refers to an RO-group wherein R is a C1-C6 alkyl group as described above; specific examples of the alkoxy group include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, sec-butoxy, n-pentoxy, isopentoxy, neopentoxy, n-hexoxy, isohexoxy, 3-methylpentoxy, 3-dimethylbutoxy, 2-ethylbutoxy and the like. "haloalkoxy" refers to a group resulting from substitution of at least one hydrogen of an alkoxy group as described above with a halogen; specific examples thereof include trifluoromethoxy and the like.

"C2-C6 alkenyloxy" refers to an RO-group, wherein R is a C2-C6 alkenyl group as described above; specific examples of the alkenyloxy group include ethyleneoxy group, propyleneoxy group.

"C2-C6 alkynyloxy" refers to an RO-group wherein R is a C2-C6 alkynyl group as described above; specific examples of alkynyloxy groups include ethynyloxy and propynyloxy.

"amino" means-NH ₂ 。

"C1-C6 alkylamino" means-NH ₂ The resulting group of which one or both hydrogens are replaced with a C1-C6 alkyl group as described above may be represented as R ₁ R ₂ N-, wherein R ₁ And R is ₂ Each independently is H or C1-C6 alkyl, and R ₁ And R is ₂ At most one is H. Specific examples of the C1-C6 alkylamino group include methylamino, dimethylamino, ethylamino, n-propylamino, isopropylamino, n-butylamino, isobutylamino, t-butylamino, sec-butylamino, n-pentylamino, isopentylamino, neopentylamino, n-hexylamino, isohexylamino, 3-methylpentylamino, 3-dimethylbutylamino, 2-ethylbutylamino and the like.

"C1-C6 Alkoxyamino" means-NH ₂ The resulting groups being substituted with C1-C6 alkyl and oxygen, respectively, as described above; specific examples of the C1-C6 alkoxyamino group include methoxyamino group, dimethoxyamino group, ethoxyamino group, n-propoxyamino group, and isopropoxy amino group.

"C3-C8 cycloalkyl" refers to a fully saturated cyclic hydrocarbon compound group containing 3-8 carbon atoms, specific examples of which include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

"C3-C6 Cycloalkylamino" means-NH ₂ The resulting group of which one or both hydrogens are replaced with a C3-C6 cycloalkyl group as described above may be represented as R ₁ R ₂ N-, wherein R ₁ And R is ₂ Each independently is H or C3-C6 cycloalkyl, and R ₁ And R is ₂ At most one isH. Specific examples of the C3-C6 cycloalkylamino group include cyclopropylamino group, cyclobutylamino group, cyclopentylamino group, cyclohexylamino group and the like.

"3-8 membered heterocyclic group" means a 3-8 membered non-aromatic cyclic alkyl group having at least one heteroatom selected from nitrogen, oxygen, sulfur in the ring; specific examples thereof include piperazine, piperidine, morpholine and the like.

"C6-C12 aryl" refers to a monocyclic or polycyclic aryl group having 6 to 12 carbon atoms; specific examples thereof include phenyl groups and naphthyl groups.

"5-to 10-membered heteroaryl" means a 5-to 10-membered aromatic group containing at least one heteroatom selected from nitrogen, oxygen, sulfur in the ring; specific examples thereof include pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyridazin-3-yl, pyridazin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, pyrazin-2-yl, pyrazin-3-yl, indolyl, isoindolyl and the like.

"pharmaceutically acceptable salts" include salts of the compounds of formula I with acids or bases; the acid comprises an inorganic acid and an organic acid; preferably, the inorganic acid comprises hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, carbonic acid; preferably, the organic acid comprises formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, citric acid, tartaric acid, carbonic acid, picric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, glutamic acid, pamoic acid; the base includes hydroxides, carbonates, bicarbonates, etc. of sodium, potassium, calcium, aluminum, lithium and ammonium.

The compounds and pharmaceutically acceptable salts thereof according to the present application may have isomers or racemates, such as optical isomers (including diastereomers and enantiomers), atropisomers, geometric isomers (cis-trans isomers), conformational isomers, tautomers, and mixtures thereof, and the like, but are not limited thereto. These isomers are also included within the scope of the present application as defined in the claims.

In another aspect, the present application provides a process for the preparation of a compound of formula I as defined above, which is carried out by the following reaction scheme:

the formula III and the formula IV are subjected to an acylation reaction in the presence of a base to obtain the formula I,

preferably, the base is selected from pyridine, sodium carbonate, sodium bicarbonate.

In yet another aspect, the present application provides a pharmaceutical composition comprising one or more selected from the group consisting of compounds of formula I and isomers, racemates, pharmaceutically acceptable salts and prodrugs thereof, and optionally pharmaceutically acceptable excipients.

In yet another aspect, the application provides the use of a compound of formula I, or an isomer, racemate, pharmaceutically acceptable salt, prodrug thereof, in the manufacture of a nav1.8 inhibitor or in the manufacture of a medicament for the treatment, prevention or management of a disease or condition associated with the nav1.8 channel.

In yet another aspect, the invention provides a method of treating, preventing or managing a disease or condition associated with nav1.8, comprising administering to a subject in need thereof one or more selected from the group consisting of a compound of formula I, an isomer, a racemate, a pharmaceutically acceptable salt and a prodrug thereof, or a pharmaceutical composition described above.

The Nav1.8 channel related diseases or symptoms include, but are not limited to, nociceptive pain, inflammatory pain, neuropathic pain, functional pain, muscle or bone injury related pain, pelvic pain, abdominal pain, chest pain, lumbosacral neuralgia, preoperative pain, postoperative pain, acute or chronic pain, migraine, trigeminal neuralgia, pancreatitis, renal colic, cancer pain, pain resulting from chemical or drug therapy, diabetic neuralgia, post-herpetic neuralgia, back pain, phantom limb pain, sciatica, small fiber neuralgia, erythromelalgia, arthritis, pruritus, acute or chronic pruritus, asthma, multiple sclerosis, arrhythmia, atrial fibrillation, heart failure, brugada syndrome, kidney stones, epilepsy, convulsions.

The invention has the following beneficial effects:

The amide compound with the structure has Nav1.8 selective inhibition activity, can be used as a Nav1.8 selective inhibitor, has better activity, higher selectivity and fewer side effects, can be used for treating, preventing or controlling diseases related to Nav1.8 channel participation or dysfunction, and has important clinical application value.

The present invention has been described in detail hereinabove, but the above embodiments are merely exemplary in nature and are not intended to limit the present invention. Furthermore, there is no intention to be bound by any theory presented in the preceding prior art or summary or the following examples.

Detailed Description

The invention is further illustrated by the following examples, which are provided for illustrative purposes only and are not to be construed as limiting the scope of the invention as claimed.

Unless otherwise indicated, all materials, reagents, methods and the like used in the examples are those conventionally used in the art.

The chinese names of the reagents represented by the chemical formulas or english abbreviations are as follows: DEG C represents DEG C; g represents gram; s represents a single peak, d represents a double peak, t represents a triple peak, and m represents a multiple peak; min represents minutes; ml stands for milliliters; mmol represents millimoles; h represents hours; TLC stands for thin layer chromatography.

In the following examples, the nuclear magnetic resonance hydrogen spectrum is recorded with a Bruker AMX-400 or AMX-600 nuclear magnetic resonance apparatus, the chemical shift δ being in ppm.

Unless otherwise indicated, all reaction solvents were purified according to conventional methods.

The thin layer chromatography uses GF254 high-efficiency plates, which are produced by the tobacco stage chemical industry research institute.

Unless otherwise noted, all solvents were analytically pure reagents, all of which were purchased from national pharmaceutical chemicals, inc.

Color development is carried out by adopting methods such as 2, 4-dinitrophenylhydrazine, iodine, ultraviolet fluorescence and the like.

The evaporation of the organic solvent under reduced pressure was carried out in a rotary evaporator.

EXAMPLE 1 Synthesis of Compound 1

[ scheme 1]

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(1) Synthesis of intermediate 1-2

Compound 1-1 (12.2 g,57.2 mmol) was dissolved in anhydrous dichloromethane (300 ml), placed in ice bath conditions, bis (2-methoxyethyl) aminotrifluorosulfur (BAST) was slowly added dropwise under nitrogen protection, after the addition was completed, the reaction was allowed to proceed to room temperature for 24h, TLC detection was essentially complete, saturated sodium bicarbonate solution was slowly added dropwise under ice bath conditions to quench, a large amount of gas was generated, extraction was performed with Dichloromethane (DCM) and water, the organic layer was dried over anhydrous sodium sulfate, and column chromatography separation (petroleum ether (PE)/Ethyl Acetate (EA) =5/1) was performed by spin-drying to give intermediate 1-2 (10 g, 74.3%). ¹ H NMR(400MHz,CDCl ₃ )δ3.51–3.38(m,3H),3.37–3.30(m,1H),2.22–1.95(m,4H),1.88–1.75(m,2H),1.46(s,9H).

(2) Synthesis of intermediates 1-3

Intermediate 1-2 (10 g,42.5 mmol) was placed in a 250ml round bottom flask, 4N dioxane solution (42.5 ml,170 mmol) was added under ice bath conditions, the reaction was completed by TLC detection at room temperature for about 4h, and the reaction solution was directly spun dry and dried to give intermediate 1-3 (7.3 g, 100%). ¹ H NMR(400MHz,DMSO)δ9.47(s,2H),3.15(m,4H),2.49–2.37(m,2H),2.30–2.13(m,2H),1.83(m,2H).

[ scheme 2]

(3) Synthesis of intermediates 1-5

Dissolving compound 1-4 (0.9 g,6.76 mmol) in 10ml concentrated sulfuric acid, transferring to ice bath condition, adding potassium nitrate (0.68 g,6.76 mmol), reacting for 3 hr under ice bath condition, TLC detecting reaction basically completely, slowly adding the reaction solution into ice water, precipitating a large amount of white solid, filtering to obtain filter cake, and drying to obtain intermediateBodies 1-5 (1.1 g, 91.3%). ¹ H NMR(400MHz,DMSO)δ8.99(s,1H),8.44(dd,J＝8.3,2.2Hz,1H),8.33(d,J＝2.0Hz,1H),7.86(d,J＝8.3Hz,1H),4.54(s,2H).

(4) Synthesis of intermediates 1-6

Intermediate 1-5 (1.1 g,6.17 mmol) was dissolved in 100ml methanol, palladium on carbon (0.22 g) with a water content of 60% was added, after 3 times of hydrogen displacement, the reaction was continued by TLC after heating to 40℃for 7h, the reaction solution was filtered and the filtrate was dried by spin-drying to give intermediate 1-6 (0.7 g, 76.6%). ¹ H NMR(400MHz,DMSO)δ8.31(s,1H),7.17(d,J＝8.0Hz,1H),6.86–6.68(m,2H),5.28(s,2H),4.16(s,2H).

[ scheme 3]

(5) Synthesis of intermediates 1-8

Intermediate 1-3 (2.38 g,12.8 mmol) was dissolved in 40ml of N, N-Dimethylformamide (DMF), potassium carbonate (5.5 g,38.5 mmol) was added, then compound 1-7 (2.0 g,11.7 mmol) was added, the reaction was heated to 120℃overnight, the reaction was checked by TLC to be substantially complete, extraction was performed with EA and water, the organic layer was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and column chromatography (PE/EA=5/1) was performed with spin-dry to give intermediate 1-8 (2.0 g, 54.9%). ¹ H NMR(400MHz,CDCl ₃ )δ7.80(d,J＝7.8Hz,1H),6.51(d,J＝7.8Hz,1H),3.85(s,3H),3.75–3.68(m,2H),3.28(m,2H),2.44–2.31(m,5H),2.01–1.89(m,4H).

(6) Synthesis of intermediates 1-9

Intermediate 1-8 (2.0 g,7.04 mmol) was dissolved in DMAC (N, N-dimethylacetamide) (40 ml), NCS (N-chlorosuccinimide) (1.8 g,14.1 mmol) was added, heated to 100℃and after 1h the reaction was complete, extracted with EA and water, the organic layer was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and column chromatography (PE/EA=10/1) was performed with spin-drying to give intermediate 1-9 (1.5 g, 66.8%). ¹ H NMR(400MHz,CDCl ₃ )δ7.84(s,1H),3.85(s,3H),3.69(dt,J＝11.9,4.4Hz,2H),3.25(t,J＝5.4Hz,2H),2.48(s,3H),2.42–2.29(m,2H),1.98–1.88(m,4H).

(7) Synthesis of intermediates 1-10

Dissolving intermediate 1-9 (1.5 g,4.7 mmol) in methanol (40 ml), adding lithium hydroxide (2.0 g,47 mmol), adding 8ml of water, heating to 50deg.C for 4h, TLC detecting reaction basically completely, spin-drying the reaction solution, adjusting pH of the reaction solution to about 3-4 with 2N hydrochloric acid, precipitating a large amount of white solid, filtering to obtain intermediate 1-10 (1.15 g, 80.3%) ¹ H NMR(400MHz,DMSO)δ12.91(s,1H),7.81(s,1H),3.65–3.55(m,2H),3.32(m,2H),2.42(s,3H),2.38–2.25(m,2H),2.04–1.93(m,2H),1.90–1.83(m,2H).

(8) Synthesis of intermediates 1-11

Intermediate 1-10 (0.05 g,0.16 mmol) was dissolved in anhydrous DCM (5 ml), the reaction was placed in ice-bath, 2-3 drops of DMF was added dropwise to catalyze, oxalyl chloride (2M, 0.1 ml) was added under nitrogen protection, and after 2h reaction in ice-bath, the reaction was spun-dried to give intermediate 1-11.

(9) Synthesis of Compound 1

Intermediate 1-6 (0.026 g,0.18 mmol) was added to intermediate 1-11 and placed in ice-bath, 3ml of anhydrous pyridine was added dropwise, after 2h TLC detection was complete, extracted with EA and water, the organic layer was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and spin-dried to column chromatography (DCM/meoh=10/1) to give compound 1 (0.015 g, 21.2%). ¹ H NMR(400MHz,DMSO)δ10.63(s,1H),8.60(s,1H),8.11(d,J＝1.6Hz,1H),7.85–7.74(m,2H),7.54(d,J＝8.2Hz,1H),5.76(s,1H),4.34(s,2H),3.62(d,J＝3.1Hz,2H),3.43(t,J＝5.8Hz,2H),2.45(s,3H),2.31(m,2H),1.94(d,J＝13.5Hz,2H),1.84(d,J＝5.5Hz,2H).

EXAMPLE 2 Synthesis of Compound 2

Referring to the procedure of step 9 of example 1, intermediate 1-6 was exchanged for 5-amino-2, 3-isoindolin-1-one (purchased from Pichia) to give compound 2 (0.021 g, 29.7%). ¹ H NMR(400MHz,DMSO)δ10.72(s,1H),8.44(s,1H),8.03(s,1H),7.76(s,1H),7.68–7.55(m,2H),4.36(s,2H),3.65–3.56(m,2H),3.44–3.37(m,6H),2.45(s,3H),2.30(s,2H),2.01–1.89(m,2H),1.86–1.77(m,2H).

EXAMPLE 3 Synthesis of Compound 3

Referring to the procedure of step 9 of example 1, intermediate 1-6 was exchanged for 6-aminoindolone (commercially available from Shaoshan) to afford compound 3 (0.020g, 29.7%). ¹ H NMR(400MHz,DMSO)δ10.45–10.38(m,2H),7.70(s,1H),7.43(s,1H),7.17–7.07(m,2H),3.64–3.58(m,2H),3.46–3.42(m,4H),2.44(s,2H),2.37–2.24(m,2H),2.03–1.90(m,2H),1.87–1.78(m,2H).

EXAMPLE 4 Synthesis of Compound 4

[ scheme 4]

(1) Synthesis of intermediate 4-2

Compound 4-1 (0.6 g,3.4 mmol) was dissolved in 50ml of methanol, palladium on carbon (0.12 g) with a water content of 60% was added to replace hydrogen 3 times, and after the reaction was continued at room temperature for 7 hours, TLC detection was complete, the reaction solution was filtered, and the filtrate was dried by spin-drying to give intermediate 4-2 (0.5 g, 100%). ¹ H NMR(400MHz,DMSO)δ9.19(s,1H),5.83–5.56(m,3H),3.90(s,2H),2.57(d,J＝2.2Hz,2H).

(2) Synthesis of Compound 4

Referring to the procedure of step 9 of example 1, intermediate 1-6 was exchanged for intermediate 4-2 to give compound 4 (0.025 g, 32.5%). ¹ H NMR(400MHz,DMSO)δ10.33(s,1H),10.28(s,1H),7.67(s,1H),7.58(s,1H),7.45–7.38(m,1H),6.77(d,J＝8.3Hz,1H),3.63–3.56(m,2H),3.48(s,2H),3.45–3.40(m,2H),2.43(s,3H),2.37–2.23(m,2H),2.03–1.91(m,2H),1.89–1.78(m,2H).

EXAMPLE 5 Synthesis of Compound 5

[ scheme 5]

(1) Synthesis of intermediate 5-2

Compound 5-1 (1.9 g,9.9 mmol) was dissolved in 10ml of absolute ethanol, and hydrazine hydrate (8)5%) (1.5 g,47.7 mmol), heated to reflux, after 2h TLC detection was complete, filtered to give a filter cake, which was dried to give crude intermediate 5-2 (1.1 g, 59.2%). ¹ H NMR(400MHz,DMSO)δ12.39(s,1H),11.13(s,1H),8.21(d,J＝1.7Hz,1H),7.86(d,J＝8.8Hz,1H),7.78(dd,J＝8.8,1.9Hz,1H).

(2) Synthesis of intermediate 5-3

Intermediate 5-2 (1.05 g,5.7 mmol) was dissolved in 60ml DCM and triethylamine (0.7 g,7.0 mmol) was added followed by di-tert-butyl dicarbonate ((Boc) ₂ O) (1.5 g,7.0 mmol), at room temperature for 3h, TLC detection was essentially complete, extraction with DCM and water, washing of the organic layer with saturated sodium chloride (NaCl), washing with anhydrous sodium sulfate (Na) ₂ SO ₄ ) Drying and spin-drying column chromatography (DCM/meoh=20/1) afforded intermediate 5-3 (1.0 g, 68.5%). ¹ H NMR(400MHz,DMSO)δ8.74(s,1H),8.04(d,J＝8.3Hz,1H),7.96(d,J＝8.6Hz,1H),1.61(s,9H).

(3) Synthesis of intermediate 5-4

Intermediate 5-3 (1.0 g,4.0 mmol) was dissolved in 50ml of methanol, palladium on carbon (0.20 g) with a water content of 60% was added, the reaction was continued for 7 hours at room temperature after 3 times of hydrogen substitution, TLC detection was complete, the reaction solution was filtered, and the filtrate was dried by spin-drying to give intermediate 5-4 (0.6 g, 89.6%). ¹ H NMR(400MHz,DMSO)δ7.30(d,J＝8.5Hz,1H),7.06(s,1H),6.52(dd,J＝8.5,1.8Hz,1H),5.86(s,2H),1.56(s,9H).

(4) Synthesis of Compound 5

Referring to step 9 of example 1, intermediate 1-6 was exchanged for intermediate 5-4 to afford intermediate 5-5. Intermediate 5-5 was dissolved in 5ml DCM, 2ml trifluoroacetic acid (TFA) was added in ice bath, reacted at room temperature for 2h, tlc detection was complete, spin-dried, EA and water were extracted, the organic layer was washed with saturated sodium bicarbonate and sodium chloride, dried over anhydrous sodium sulfate, and spin-dried column chromatography (DCM/meoh=10/1) gave compound 5 (0.05 g, 33.1%). ¹ H NMR(400MHz,MeOD)δ8.06–8.03(m,1H),7.78–7.71(m,2H),7.66–7.60(m,1H),7.13(dd,J＝8.6,1.6Hz,1H),3.78–3.68(m,2H),3.49(t,J＝5.9Hz,2H),2.51(s,3H),2.35(td,J＝10.6,5.5Hz,2H),2.07–1.98(m,2H),1.98–1.87(m,2H),1.78–1.68(m,2H),1.47(dt,J＝14.7,7.5Hz,2H).

EXAMPLE 6 Synthesis of Compound 6

[ scheme 6]

(1) Synthesis of intermediate 6-2

Compound 6-1 (0.6 g,2.6 mmol) was dissolved in 30ml of methanol, palladium on carbon (0.12 g) having a water content of 60% was added thereto, the reaction was continued for 7 hours at room temperature after 3 times of replacement of hydrogen, and after completion of the TLC detection, the reaction solution was filtered and the filtrate was dried by spinning to obtain intermediate 6-2 (0.4 g, 76.7%). ¹ H NMR(400MHz,DMSO)δ7.59(d,J＝8.5Hz,1H),6.94(d,J＝1.9Hz,1H),6.89(dd,J＝8.5,2.0Hz,1H),6.77(s,1H).

(2) Synthesis of intermediate 6-3

Intermediate 6-2 (0.4 g,2.0 mmol) was dissolved in 5ml of concentrated hydrochloric acid under ice-bath conditions, zinc powder (1.1 g,16.2 mmol) was added in portions, the reaction was naturally warmed to room temperature after completion of the addition, after 2h, TLC detection was complete, the reaction solution was slowly poured into ice water to quench, pH was adjusted to around 7 with 2N sodium hydroxide, extracted with EA and water, the organic layer was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and column chromatography (DCM/meoh=10/1) was performed by spin-drying to give intermediate 6-3 (0.10 g, 27.1%). ¹ H NMR(400MHz,DMSO)δ7.55–7.52(m,1H),7.14(d,J＝8.3Hz,1H),6.83(dd,J＝8.3,2.1Hz,1H),6.79(d,J＝1.9Hz,1H),5.60(s,2H),4.18(d,J＝4.7Hz,2H).

(3) Synthesis of Compound 6

Referring to step 9 of example 1, intermediate 1-6 was exchanged for intermediate 6-3 to give compound 6 (0.048 g, 37.3%). ¹ H NMR(600MHz,DMSO)δ10.79(s,1H),8.19(s,1H),7.86(t,J＝4.6Hz,1H),7.84–7.77(m,2H),7.54(d,J＝8.4Hz,1H),4.37(d,J＝4.5Hz,2H),3.61(dd,J＝6.5,3.6Hz,2H),3.40(t,J＝5.9Hz,2H),2.45(s,3H),2.31(m,2H),2.01–1.92(m,2H),1.84(d,J＝5.5Hz,2H).

EXAMPLE 7 Synthesis of Compound 7

[ scheme 7]

(1) Synthesis of intermediate 7-2

Compound 7-1 (1.0 g,5.6 mmol) was dissolved in 40ml of methanol, palladium on carbon (0.2 g) having a water content of 60% was added thereto, the reaction was continued for 7 hours at room temperature after 3 times of replacement of hydrogen, and after completion of the TLC detection, the reaction solution was filtered and the filtrate was dried by spinning to obtain intermediate 7-2 (0.7 g, 83.2%). ¹ H NMR(400MHz,DMSO)δ11.05(s,1H),6.74(d,J＝8.3Hz,1H),6.50(d,J＝2.0Hz,1H),6.35(dd,J＝8.3,2.0Hz,1H),5.00(s,2H).

(2) Synthesis of Compound 7

Referring to step 9 of example 1, intermediate 1-6 was exchanged for intermediate 7-2 to give compound 7 (0.018 g, 28.5%). ¹ H NMR(400MHz,MeOD)δ7.74(d,J＝1.9Hz,1H),7.68(s,1H),7.34(dd,J＝8.4,2.0Hz,1H),7.05(d,J＝8.4Hz,1H),3.73–3.66(m,2H),3.48(t,J＝5.9Hz,2H),2.48(s,3H),2.39–2.26(m,2H),1.99–1.86(m,4H).

EXAMPLE 8 Synthesis of Compound 8

[ scheme 8]

(1) Synthesis of intermediate 8-2

Compound 8-1 (2.0 g,9.9 mmol) was dissolved in 20ml absolute ethanol, hydrazine hydrate (85%) (2.92 g,49.6 mmol) was added, heated to reflux, TLC detection after 3h was complete, filtered to give a filter cake, and the filter cake was dried to give crude product 8-2 (2.0 g, 102.24%). ¹ H NMR(400MHz,DMSO)δ12.45(s,1H),8.67(d,J＝2.0Hz,1H),8.11(dd,J＝9.2,2.2Hz,1H),7.45(d,J＝9.2Hz,1H).

(2) Synthesis of intermediate 8-3

Dissolving crude product 8-2 (2.0 g,10.1 mmol) in 30ml water, adding 3ml concentrated hydrochloric acid, heating to 90deg.C, reacting overnight, detecting the reaction by TLC to obtain a filter cake, filtering, and drying the filter cakeDrying gave crude intermediate 8-3 (1.67 g, 91.9%). ¹ H NMR(400MHz,DMSO)δ12.43(s,1H),8.67(d,J＝1.9Hz,1H),8.12(dd,J＝9.2,2.2Hz,1H),7.45(d,J＝9.2Hz,1H).

(3) Synthesis of intermediate 8-4

Intermediate 8-3 (1.5 g,8.4 mmol) was dissolved in 50ml DCM and triethylamine (2.5 g,25.1 mmol) was added followed by (Boc) ₂ O (2 g,9.2 mmol), at room temperature for 3h, TLC detection was essentially complete, extraction with DCM and water, washing the organic layer with saturated NaCl, washing with anhydrous Na ₂ SO ₄ Drying and spin-drying column chromatography (DCM/meoh=20/1) gave intermediate 8-4 (1.5 g, 64.1%). ¹ H NMR(400MHz,DMSO)δ11.66(s,1H),7.64(s,1H),6.88(d,J＝8.7Hz,1H),6.73(s,1H),1.57(s,9H).

(4) Synthesis of intermediate 8-5

Intermediate 8-4 (1.5 g,5.4 mmol) was dissolved in 50ml of methanol, palladium on carbon (0.30 g) with a water content of 60% was added, the reaction was continued for 7 hours at room temperature after 3 times of hydrogen substitution, TLC detection was complete, the reaction solution was filtered, and the filtrate was dried by spin-drying to give intermediate 8-5 (1.2 g, 89.6%). ¹ H NMR(400MHz,DMSO)δ7.64(d,J＝7.9Hz,1H),6.88(dd,J＝8.8,2.2Hz,1H),6.73(d,J＝2.0Hz,1H),5.18(s,2H),1.56(s,9H).

(5) Synthesis of Compound 8

Referring to step 9 of example 1, intermediate 1-6 was exchanged for intermediate 8-5 to afford intermediate 8-6 (0.091 g, 34.6%); intermediate 8-6 was dissolved in DCM (5 ml), 2ml TFA was added in ice bath, the reaction was completed by tlc at room temperature, the reaction was completed, spin-dried, EA and water were extracted, the organic layer was washed with saturated sodium bicarbonate and sodium chloride, dried over anhydrous sodium sulfate, and spin-dried column chromatography (DCM/meoh=10/1) to give compound 8 (0.05 g, 25.5%). ¹ H NMR(400MHz,DMSO)δ11.28(s,1H),10.56(s,1H),10.37(s,1H),8.08(s,1H),7.72(s,1H),7.46(m,1H),7.26(d,J＝8.9Hz,1H),3.62(m,2H),3.46(t,J＝5.8Hz,2H),2.44(s,3H),2.30(m,2H),1.96(m,2H),1.84(m,2H).

EXAMPLE 9 Synthesis of Compound 9

Referring to step 9 of example 1, intermediate 1-6 was exchanged for intermediate 6-2,compound 9 (9 mg, 19.8%) was obtained. ¹ H NMR(400MHz,DMSO)δ7.92(s,1H),7.74(d,J＝8.7Hz,1H),7.20(s,1H),7.08(d,J＝1.9Hz,1H),7.00(dd,J＝8.7,1.9Hz,1H),3.60(m,2H),3.20(m,2H),2.46(s,3H),2.29(m,2H),1.90(m,4H).

EXAMPLE 10 Synthesis of Compound 10

Referring to step 9 of example 1, intermediate 1-6 was replaced with 4-aminophthalhydrazide (purchased from Lev.) to give compound 10 (8.0 mg, 22.5%). ¹ H NMR(400MHz,MeOD)δ8.60(s,1H),8.17(d,J＝14.0Hz,1H),8.13(d,J＝7.5Hz,1H),7.76(s,1H),3.73(d,J＝5.0Hz,2H),3.47(t,J＝5.8Hz,2H),2.50(s,3H),2.27-2.40(m,2H),2.03–1.86(m,4H).

EXAMPLE 11 Synthesis of Compound 11

[ scheme 9]

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(1) Synthesis of intermediate 11-2

Referring to step 9 of example 1, intermediate 1-6 was exchanged for compound 11-1 to give intermediate 11-2 (0.48 g, 69.1%). ¹ H NMR(400MHz,DMSO)δ10.77(s,1H),8.20–8.16(m,1H),7.94(s,1H),7.79(s,1H),7.55(t,J＝9.1Hz,1H),3.60(m,2H),3.40–3.35(m,2H),2.44(s,3H),2.25-2.35(m,2H),1.91-1.98(m,2H),1.80-1.87(m,2H).

(2) Synthesis of Compound 11

Intermediate 11-2 (0.12 g,0.28 mmol) was dissolved in ultra-dry DMF (3 ml), potassium tert-butoxide (0.048 g,0.426 mmol) and acetohydroxamic acid (0.032 g,0.43 mmol) were added, after reaction for 4h at 50 ℃, TLC detection was complete, extracted with EA and water, the organic layer was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and column chromatography (DCM/meoh=20/1) was performed with spin-drying to give compound 11 (0.035 g, 28.3%). ¹ H NMR(600MHz,DMSO)δ10.55(s,1H),8.27(d,J＝1.7Hz,1H),7.73(s,1H),7.58(dd,J＝8.9,2.0Hz,1H),7.43(d,J＝8.9Hz,1H),6.40(s,2H),3.64–3.61(m,2H),3.46(t,J＝6.1Hz,2H),2.45(s,3H),2.31(2.26-2.35,2H),1.91-97(m,2H),1.86–1.82(m,2H).

EXAMPLE 12 Synthesis of Compound 12

[ scheme 10]

Intermediate 11-2 (0.10 g,0.28 mmol) was dissolved in ethanol (10 ml), 0.3ml of hydrazine hydrate solution was added, the reaction was heated to 80 ℃ and refluxed overnight, the next day TLC detection was complete, extracted with EA and water, the organic layer was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and spin-dried to column chromatography (DCM/meoh=20/1) to give compound 12 (0.067 g, 65.1%). ¹ H NMR(400MHz,DMSO)δ11.34(s,1H),10.31(s,1H),8.05(s,1H),7.69(s,1H),7.35(d,J＝8.6Hz,1H),7.20(d,J＝8.7Hz,1H),5.29(s,2H),3.61-3.66(m,2H),3.50–3.46(m,2H),2.44(s,3H),2.25-2.37(m,2H),1.93-2.03(m,2H),1.82-1.87(m,2H).

EXAMPLE 13 Synthesis of Compound 13

[ scheme 11]

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(1) Synthesis of intermediate 13-1

Compound 2, 6-dichloronicotinic acid (2.0 g,10.4 mmol) and intermediate 1-3 (2.1 g,12.5 mmol) were dissolved in DMF (100 ml), potassium carbonate (7.0 g,52 mmol) was added, after heating the reaction solution to 80℃for 4h, the reaction was checked to be substantially complete by TLC, quenched with water, pH was adjusted to around 4 with 2M hydrochloric acid, extracted with EA and water, the organic layer was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and column chromatographed on dry (DCM/MeOH=20/1) to give intermediate 13-1 (2.57 g, 85%). ¹ H NMR(400MHz,Chloroform-d)δ8.25(t,J＝8.3Hz,1H),6.35(dd,J＝8.3Hz,1H),3.69–3.65(m,2H),3.37(m,2H),2.40(m,2H),2.07–2.01(m,2H),1.98(m,2H).

(2) Synthesis of intermediate 13-2

Compound 13-1 (0.5 g,1.72 mmol) was dissolved in anhydrous dichloromethane (40 ml) and placed under ice-bath conditions under nitrogen protectionOxalyl chloride (0.18 ml,2.1 mmol) is added dropwise, and a catalytic amount of DMF is added, and the reaction is carried out for about 2 hours under the ice bath condition, and the reaction solution is directly dried by spinning, so as to obtain a crude product and placed in a reaction bottle; to this flask was added 5-amino-2-fluorobenzonitrile and anhydrous pyridine (5 ml) was added dropwise under nitrogen protection and ice-bath conditions, after 2h TLC detection was essentially complete, extracted with EA and water, the organic layer was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and spin-dried to give intermediate 13-2 (0.4 g, 56%). ¹ H NMR(400MHz,DMSO)δ10.77(s,1H),8.19(d,J＝5.8,1H),7.95(m,1H),7.90(m,1H),7.55(t,J＝9.2Hz,1H),6.45(dd,J＝8.0,3.1Hz,1H),3.57(s,2H),3.42–3.37(m,2H),2.31(s,2H),1.96(s,2H),1.86(d,J＝5.0Hz,2H).

(3) Synthesis of intermediate 13-3

Intermediate 13-2 (0.4 g,0.98 mmol), potassium tetrafluorocyclopropane (0.36 g,2.5 mmol) and potassium carbonate (0.41 g,2.9 mmol) were dissolved in dioxane/water (10 ml/2 ml) and after 3-4 times aeration with nitrogen Pd (dppf) was added ₂ Cl ₂ (0.07 g,0.098 mmol) and heating the reaction to 100deg.C overnight, TLC detection was essentially complete, extraction with EA and water, washing of the organic layer with saturated sodium chloride, drying over anhydrous sodium sulfate, and column chromatography on a spin-dry agent (DCM/MeOH=50/1) afforded intermediate 13-3 (0.1 g, 25%). ¹ H NMR(400MHz,DMSO)δ10.22(s,1H),8.19(m,1H),7.95(m,1H),7.90(m,1H),7.55(t,J＝9.2Hz,1H),6.64(d,J＝8.0,1H),3.57(s,2H),3.42–3.37(m,2H),2.35–2.26(m,2H),2.05–1.91(m,2H),1.88–1.81(m,2H).

(4) Synthesis of intermediate 13-4

Intermediate 13-3 (0.1 g,0.24 mmol) was dissolved in 2ml of DMAC solution, NCS (35 mg,0.26 mmol) was added, the reaction was allowed to react for 0.5h at 100deg.C, TLC was monitored to be essentially complete, extracted with EA and water, the organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and column chromatographed after spin-drying (DCM/MeOH=20:1) to give intermediate 13-4 (85 mg, 78%). ¹ H NMR(400MHz,DMSO)δ10.22(s,1H),8.17(s,1H),7.88(m,2H),7.48(t,J＝9.2Hz,1H),3.57(m,2H),3.45–3.39(m,2H),2.38–2.29(m,2H),2.15–2.03(m,2H),1.88–1.85(m,2H),1.24(m,2H),0.99(m,2H).

(5) Synthesis of Compound 13

Referring to step (2) of example 11, intermediate 11-2 was exchanged for intermediate 13-4 to give compound 13 (8 mg, 18.3%). ¹ H NMR(400MHz,DMSO-d ₆ )δ10.73(s,1H),7.97(s,1H),7.77(s,1H),7.73(d,J＝8.5Hz,1H),7.40–7.35(d,J＝8.5Hz,1H),6.35(s,2H),3.61(m,2H),3.40(t,2H),2.45(s,3H),2.30(m,2H),1.93(m,2H),1.83(m,2H)1.23(m,2H),0.98(m,2H).

EXAMPLE 14 Synthesis of Compound 14

[ scheme 12]

(1) Synthesis of intermediate 14-1

Intermediate 13-2 (0.2 g,0.49 mmol) was dissolved in 1ml of DMSO solution and 2ml of methanol and K were added ₂ CO ₃ (0.06 g,0.49 mmol) was reacted for 1h at 80℃with microwaves, TLC monitored that the reaction was essentially complete, extracted with EA and water, the organic phase was washed with saturated sodium chloride and dried over anhydrous sodium sulfate, and column chromatography after spin-drying (DCM/MeOH=20:1) afforded intermediate 14-1 (0.09 g, 45%). ¹ H NMR(600MHz,DMSO)δ10.56(s,1H),8.19(dd,J＝5.7,2.8Hz,1H),7.94(m,1H),7.68(d,J＝8.2Hz,1H),7.53(d,J＝9.1Hz,1H),6.16(d,J＝8.2Hz,1H),3.84(s,3H),3.63–3.60(m,2H),3.42–3.39(m,2H),2.39–2.33(m,2H),1.98(m,2H),1.85(m,2H).

(2) Synthesis of intermediate 14-2

Referring to step (4) of example 13, intermediate 13-3 was changed to 14-1 to give intermediate 14-2 (0.06 g, 64%). ¹ H NMR(400MHz,DMSO)δ10.66(s,1H),8.18(dd,J＝5.7,2.8Hz,1H),7.93(m,1H),7.85(s,1H),7.55(d,J＝9.1Hz,1H),3.94(s,3H),3.61(m,2H),3.37(m,2H),2.37–2.31(m,2H),1.96(m,2H),1.86(m,2H).

(3) Synthesis of Compound 14

Referring to step (2) of example 11, intermediate 11-2 was exchanged for intermediate 14-2 to give product 14 (11 mg, 20.8%). ¹ H NMR(400MHz,DMSO)δ10.45(s,1H),8.27(m,1H),7.79(s,1H),7.59(d,J＝8.0Hz,1H),7.43(d,J＝8.8Hz,1H),6.41(s,2H),3.93(s,3H),3.65(m,2H),3.46(m,2H),2.33(m,2H),1.99(m,2H),1.87(m,2H).

EXAMPLE 15 Synthesis of Compound 15

Referring to step (1) of example 11, intermediate 11-1 was exchanged for 4-amino-2-fluorobenzonitrile (purchased after that) to afford intermediate 15-1. ¹ H NMR(400MHz,DMSO-d ₆ )δ11.05(s,1H),7.91(s,1H),7.82(s,1H),7.65(t,J＝8.2Hz,1H),7.57(dd,J＝8.6,1.9Hz,1H),3.60(m,2H),3.08(m,2H),2.45(s,3H),2.31(m,2H),1.90-1.96(d,2H),1.80-1.85(m,2H).

Referring to step (2) of example 11, intermediate 11-2 was exchanged for intermediate 15-1 to give compound 15. ¹ H ¹ H NMR(400MHz,DMSO-d ₆ )δ10.73(s,1H),7.97(s,1H),7.77(s,1H),7.73(d,J＝8.5Hz,1H),7.40–7.35(d,J＝8.5Hz,1H),6.35(s,2H),3.61(m,2H),3.40(t,2H),2.45(s,3H),2.30(m,2H),1.93(m,2H),1.83(m,2H).

EXAMPLE 16 Synthesis of Compound 16

Referring to the procedure of example 12, intermediate 11-2 was exchanged for intermediate 15-1 to give compound 16.

¹ H NMR(400MHz,DMSO-d ₆ )δ11.28(s,1H),10.45(s,1H),7.89(s,1H),7.72(s,1H),7.58(d,J＝8.6Hz,1H),7.02–6.97(dd,J＝8.6Hz,1H),5.29(s,2H),3.61(m,2H),3.44(t,2H),2.44(s,3H),2.30(m,2H),1.95(m,2H),1.83(m,2H).

EXAMPLE 17 Synthesis of Compound 17

[ scheme 13]

(1) Synthesis of intermediate 17-2

Intermediate 17-1 (0.5 g,2.7 mmol) was dissolved in 20ml of methanol, palladium on carbon (0.1 g) with a water content of 60% was added to replace hydrogen 3 times, and after the reaction was continued at room temperature for 7 hours, the reaction was completed by TLC, and the reaction solution was filtered and the filtrate was dried by spin-drying to give intermediate 17-2 (0.093 g, 22.4%). ¹ H NMR(400MHz,DMSO)δ7.96(d,J＝2.7Hz,1H),7.39(d,J＝2.7Hz,1H),6.03(s,2H).

(2) Synthesis of intermediate 17-3

Referring to step (1) of example 11, intermediate 11-1 was exchanged for intermediate 17-2 to give intermediate 17-3 (0.11 g, 29.6%). ¹ H NMR(600MHz,MeOD)δ8.81(d,J＝1.7Hz,1H),8.67(d,J＝1.9Hz,1H),7.78(s,1H),3.72–3.68(m,2H),3.41(q,J＝5.1Hz,2H),2.50(s,3H),2.33(td,J＝10.5,5.3Hz,2H),1.99–1.90(m,4H).

(3) Synthesis of Compound 17

Referring to step (2) of example 11, intermediate 11-2 was exchanged for intermediate 17-3 to give compound 17 (9 mg, 19.8%). ¹ H NMR(600MHz,CDCl ₃ )δ10.48(s,1H),8.83(d,J＝2.3Hz,1H),8.41(d,J＝2.4Hz,1H),8.14(s,1H),4.61(s,2H),3.59–3.56(m,2H),3.39(t,J＝5.7Hz,2H),2.57(s,3H),2.41(ddd,J＝15.3,9.9,5.5Hz,2H),2.21–2.15(m,2H),1.94(dd,J＝11.3,5.6Hz,2H).

EXAMPLE 18 Synthesis of Compound 18

Referring to example 12, intermediate 11-2 was exchanged for intermediate 17-3 to give compound 18 (25 mg, 53%). ¹ H NMR(400MHz,DMSO)δ11.92(s,1H),10.51(s,1H),8.49(d,J＝2.2Hz,1H),8.43(d,J＝2.3Hz,1H),7.77(s,1H),5.57(s,2H),3.66–3.60(m,2H),3.49–3.44(m,2H),2.45(s,3H),2.31(d,J＝15.8Hz,2H),1.98(d,J＝7.5Hz,2H),1.86(d,J＝5.3Hz,2H).

EXAMPLE 19 Synthesis of Compound 19

Referring to step (1) of example 11, intermediate 11-1 was replaced with intermediate 6-amino-2-chlorocyanopyridine to give intermediate 19-1. ¹ H NMR(400MHz,DMSO)δ11.92(s,1H),8.72(d,J＝14.9Hz,1H),8.23(d,J＝15.1Hz,1H),7.79(s,1H),3.72–3.68(m,2H),3.41(3.49-3.46,m,2H),2.51(s,3H),2.01-1.98(m,2H),1.99–1.90(m,4H).

Referring to step (2) of example 11, intermediate 11-2 was exchanged for intermediate 19-1 to give compound 19. ¹ H NMR(400MHz,CDCl ₃ )δ11.67(s,1H),8.24(s,1H),7.86(d,J＝15.1Hz,1H),7.71(d,J＝15.3Hz,1H),6.18(s,2H),3.58–3.55(m,2H),3.39(m,2H),2.45(s,3H),2.39-2.37(m,2H),2.21–2.15(m,2H),1.92-1.89(m,2H).

EXAMPLE 20 Synthesis of Compound 20

Referring to example 12, intermediate 11-2 was exchanged for intermediate 19-1 to give compound 20. ¹ H NMR(400MHz,DMSO)δ12.12(s,1H),11.01(s,1H),8.26(s,1H),7.87(d,J＝15.3Hz,1H),7.43(d,J＝14.9Hz,1H),5.28(s,2H),3.69–3.64(m,2H),3.49–3.44(m,2H),2.51(s,3H),2.31-2.29(m,2H),1.98-1.95(m,2H),1.86-1.82(m,2H).

EXAMPLE 21 Synthesis of Compound 21

[ scheme 14]

(1) Synthesis of intermediate 21-1

5-bromo-2-hydrazinopyridine (2 g,10.6 mmol) was dissolved in tetrahydrofuran, N' -carbonyldiimidazole (2.6 g,15.9 mmol) was added at room temperature, and the reaction mixture was heated to reflux overnight; the reaction was essentially complete as detected by TLC, extraction with ethyl acetate and water, washing the organic layer with saturated sodium chloride and drying over anhydrous sodium sulfate, and spin-drying afforded crude intermediate 21-1 (1.3 g, 58%).

(2) Synthesis of intermediate 21-2

Intermediate 21-1 (1.3 g,6.07 mmol) was dissolved in 15ml of phosphorus oxychloride, heated to 110 ℃ and reacted overnight, TLC detection was essentially complete, the reaction cooled and slowly added dropwise to ice water and extracted with ethyl acetate and water, the organic layer washed with saturated sodium bicarbonate, saturated sodium chloride, dried over anhydrous sodium sulfate, and spin-dried to afford intermediate 21-2 (0.86 g, 61%). ¹ H NMR(400MHz,CDCl ₃ )δ8.17(s,1H),7.67(d,J＝9.7Hz,1H),7.38(dd,J＝9.7,1.6Hz,1H).

(3) Synthesis of intermediate 21-3

Compound 21-2 (0.86 g,3.69 mmol) was dissolved in benzylamine (5 ml), reacted overnight at 110℃and the reaction solution was extracted with ethyl acetate and water, the organic layer was washed with saturated sodium chloride, dried over anhydrous sodium sulfate and spin-dried to give intermediate 21-3 by column chromatography (DCM/MeOH=20/1).

(4) Synthesis of intermediate 21-4

Dissolving the intermediate 21-3 in DMSO, adding potassium carbonate, L-proline and cuprous iodide, stirring the reaction solution under the condition of nitrogen protection for 5 minutes at room temperature, adding ammonia water, heating the reaction solution to 90 ℃ overnight, performing TLC detection on the reaction basically completely, extracting with ethyl acetate and water, washing an organic layer with saturated sodium chloride, drying with anhydrous sodium sulfate, spin-drying, and separating by column chromatography to obtain the intermediate 21-4.

(5) Synthesis of intermediate 21-5

Compound 21-4 was dissolved in 50ml of methanol, palladium carbon with a water content of 60% was added to replace hydrogen for 3 times, and after the reaction was continued for 7 hours at room temperature, TLC detection was completed, the reaction solution was filtered, and the filtrate was dried by spin-drying to obtain intermediate 21-5.

(6) Synthesis of Compound 21

Referring to step (9) of example 1, intermediate 1-6 was exchanged for intermediate 21-5 to give compound 21.

EXAMPLE 22 Synthesis of Compound 22

Referring to step (9) of example 1, intermediates 1 to 6 were exchanged for 4-aminophthalimide to give compound 22. ¹ H NMR(400MHz,DMSO)δ11.26(s,1H),10.99(s,1H),8.23(s,1H),7.97(d,J＝8.0Hz,1H),7.84–7.79(m,2H),3.61(m,2H),3.38(m,2H),2.45(s,3H),2.32(m,2H),2.02–1.97(m,2H),1.84(d,J＝5.6Hz,2H).

EXAMPLE 23 Synthesis of Compound 23

[ scheme 15]

(1) Synthesis of intermediate 23-1

4-chloro-7-nitroquinazoline (0.5 g,2.39 mmol) was dissolved in 20ml of ammonia in methanol (7N) and reacted at room temperature for 2h, the reaction was completed by TLC, and the reaction solution was dried by spin-drying to give a crude product 23-1 (0.3 g). ¹ H NMR(400MHz,DMSO)δ8.52(m,2H),8.38(d,J＝2.1Hz,1H),8.20(dd,J＝9.0,2.2Hz,1H),7.30(brs,2H).

(2) Synthesis of intermediate 23-2

Intermediate 23-1 (0.3 g,0.9 mmol) was dissolved in 30ml of methanol, palladium on carbon (0.06 g) having a water content of 60% was added thereto, the reaction was continued for 7 hours at room temperature after 3 times of replacement of hydrogen, and after completion of the reaction by TLC, the reaction solution was filtered and the filtrate was dried by spin-drying to obtain intermediate 23-2. ¹ H NMR(400MHz,DMSO)δ8.21(s,1H),7.88(d,J＝8.9Hz,1H),7.61(s,2H),6.78(dd,J＝8.9,2.2Hz,1H),6.59(d,J＝2.2Hz,1H),6.11(s,2H).

(3) Synthesis of Compound 23

Referring to step (9) of example 1, intermediate 1-6 was changed to intermediate 23-2 to give compound 23 (12 mg, 11%). ¹ H NMR(400MHz,CDCl ₃ )δ10.14(s,1H),8.60(s,1H),8.15(s,1H),8.06(d,J＝8.2Hz,1H),7.90(s,1H),7.78(d,J＝9.0Hz,1H),5.71(s,2H),3.61–3.57(m,2H),3.45–3.40(m,2H),2.59(s,3H),2.46–2.36(m,2H),2.23–2.12(m,2H),1.96–1.90(m,2H).

EXAMPLE 24 Synthesis of Compound 24

Referring to the synthetic method of example 23, 4-chloro-7-nitroquinazoline was exchanged for 4-chloro-6-nitroquinazoline to afford compound 24 (61 mg, 56%). ¹ H NMR(400MHz,CDCl ₃ )δ10.41(s,1H),8.64(d,J＝2.1Hz,1H),8.60(s,1H),8.19(s,1H),7.88(d,J＝9.0Hz,1H),7.64–7.60(m,1H),5.78(s,2H),3.62–3.58(m,2H),3.41–3.37(m,2H),2.58(s,1H),2.45(ddd,J＝14.7,10.3,5.1Hz,2H),2.26–2.16(m,2H),1.97–1.91(m,2H).

EXAMPLE 25 Synthesis of Compound 25

[ scheme 16]

(1) Synthesis of intermediate 25-1

4-chloro-7-nitroquinazoline (0.5 g,2.39 mmol) was dissolved in methanol (20 ml), sodium hydride (60% in kerosene) was added under ice bath (0.14 g,3.6 mmol), reacted overnight at room temperature, the reaction was essentially complete as detected by TLC, and ice-water was added Quench, extract with EA and water, wash the organic layer with saturated sodium bicarbonate and sodium chloride, dry over anhydrous sodium sulfate, and spin-dry column chromatography (DCM/meoh=20/1) to give intermediate 25-1 (0.4 g, 81.6%). ¹ H NMR(400MHz,DMSO)δ9.00(s,1H),8.67(s,1H),8.43–8.34(m,2H),4.20(s,3H).

(2) Synthesis of intermediate 25-2

Intermediate 25-1 (0.4 g,1.94 mmol) was dissolved in 50ml of methanol, palladium on carbon (0.08 g) with a water content of 60% was added, the reaction was continued for 7 hours at room temperature after 3 times of hydrogen substitution, TLC detection was complete, the reaction solution was filtered, and the filtrate was dried by spin-drying to give intermediate 25-2 (0.3 g, 91.2%). ¹ H NMR(400MHz,DMSO)δ8.47(s,1H),7.78(d,J＝8.8Hz,1H),6.93(dd,J＝8.9,2.2Hz,1H),6.76(d,J＝2.1Hz,1H),6.21(s,2H),4.01(s,3H).

(3) Synthesis of intermediate 25-3

Referring to step (9) of example 1, intermediate 1-6 was exchanged for intermediate 25-2 to give intermediate 25-3 (0.09 g, 61%).

(4) Synthesis of Compound 25

Intermediate 25-3 (0.09 g,0.2 mmol) was dissolved in DMF (5 ml), lithium chloride (0.04 g,1.0 mmol) and p-toluenesulfonic acid (0.17 g,1.0 mmol) were added, the reaction mixture was heated to 80℃for about 3h, the reaction was essentially complete by TLC detection, cooled, extracted with EA and water, the organic layer was washed with saturated sodium bicarbonate and sodium chloride, dried over anhydrous sodium sulfate, and spin-dried column chromatography (DCM/MeOH=10/1) to give compound 25 (33 mg, 39%). ¹ H NMR(400MHz,DMSO)δ12.15(s,1H),10.84(s,1H),8.13–8.04(m,3H),7.81(s,1H),7.75–7.70(m,1H),3.62(s,2H),3.40(t,J＝5.8Hz,2H),2.45(s,3H),2.32(s,2H),1.99–1.90(m,2H),1.84(d,J＝5.2Hz,2H).

EXAMPLE 26 Synthesis of Compound 26

[ scheme 17]

(1) Synthesis of intermediate 26-1

3-hydroxy-1, 2-benzeneAnd isoxazole (obtained after purchase) (1.0 g,7.4 mmol) was dissolved in 10ml of concentrated sulfuric acid, and the solution was transferred to ice bath conditions, potassium nitrate (0.74 g,7.4 mmol) was added, the reaction was carried out for 3 hours under ice bath conditions, TLC detection was substantially complete, the reaction solution was slowly added to ice water, a large amount of white solid was precipitated, and a cake was obtained by filtration, and intermediate 26-1 (1.3 g, 97.5%) was obtained by drying. ¹ H NMR(400MHz,DMSO)δ13.05(s,1H),8.68(d,J＝2.1Hz,1H),8.46(dd,J＝9.2,2.3Hz,1H),7.82(dd,J＝9.2,4.6Hz,1H).

(2) Synthesis of intermediate 26-2

Intermediate 26-1 (0.4 g,2.2 mmol) was dissolved in 30ml of dichloromethane, and the reaction mixture was placed in an ice-salt bath, 2- (trimethylsilyl) ethoxymethyl chloride (0.74 g,4.4 mmol) was added under nitrogen protection, triethylamine (0.67 g,6.7 mmol) was slowly added dropwise after stirring for 5min, after ice-salt bath reaction for 2h, TLC detection was complete, extraction was performed with DCM and water, the organic layer was washed with saturated sodium chloride and dried over anhydrous sodium sulfate, and column chromatography separation was performed by spin-drying (PE/ea=10/1) to give intermediate 26-2 (0.28 g, 41%). ¹ H NMR(400MHz,CDCl ₃ )δ8.79(d,J＝2.0Hz,1H),8.55(dd,J＝9.2,2.3Hz,1H),7.41(d,J＝9.2Hz,1H),5.40(s,2H),3.75–3.64(m,2H),1.00–0.91(m,2H),-0.01(s,9H).

(3) Synthesis of intermediate 26-3

Intermediate 26-2 (0.28 g,0.9 mmol) was dissolved in 100ml methanol, palladium on carbon (0.05 g) with a water content of 60% was added to replace hydrogen 3 times, and after continued reaction at room temperature for 1h, TLC detection was complete, the reaction solution was filtered, the filtrate was dried by spin-drying, and column chromatography (DCM/MeOH=20/1) gave intermediate 26-3 (0.07 g, 28%). ¹ H NMR(400MHz,CDCl ₃ )δ7.05(dd,J＝9.1,5.4Hz,2H),7.00(dd,J＝8.7,2.4Hz,1H),5.32(s,2H),3.71–3.64(m,2H),0.98–0.92(m,2H),-0.01(s,9H).

(4) Synthesis of intermediate 26-4

Referring to step (9) of example 1, intermediate 1-6 was exchanged for intermediate 26-3 to obtain intermediate 26-4. ¹ H NMR(400MHz,CDCl ₃ )δ9.78(s,1H),8.02(d,J＝3.2Hz,2H),7.23(d,J＝3.9Hz,1H),5.31(s,2H),3.67–3.61(m,2H),3.59–3.54(m,2H),3.41–3.36(m,2H),2.51(s,3H),2.34(td,J＝10.3,5.8Hz,2H),2.14–2.05(m,2H),1.93–1.86(m,2H),0.96–0.89(m,2H),-0.04(s,9H).

(5) Synthesis of Compound 26

Intermediate 26-4 (50 mg,0.09 mmol) was dissolved in dichloromethane, 1ml of trifluoroacetic acid was added under ice-bath conditions, and after stirring at room temperature for 1h, the reaction was complete by TLC detection and the reaction solution was spin-dried for column chromatography (DCM/meoh=20/1) to give compound 26 (10 mg, 25.4%). ¹ H NMR(600MHz,DMSO)δ12.36(s,1H),10.62(s,1H),8.23(d,J＝1.6Hz,1H),7.77(s,1H),7.71(d,J＝9.1Hz,1H),7.54(d,J＝9.0Hz,1H),3.62(s,2H),3.44(t,J＝6.1Hz,2H),3.17(d,J＝4.5Hz,1H),2.45(s,3H),2.31(s,2H),1.96(s,2H),1.84(d,J＝5.5Hz,2H).

EXAMPLE 27 Synthesis of Compound 27

[ scheme 18]

(1) Synthesis of intermediate 27-1

3, 6-dichloropyridazine-4-carboxylic acid (0.5 g,2.59 mmol), compounds 1-3 (0.534 g,3.11 mmol), potassium carbonate (1.79 g,12.95 mmol) were dissolved in 30ml DMF and reacted overnight at 80 ℃. After completion of the TLC detection reaction, the reaction solution was acidified with 3N HCl solution, extracted with EA and water, the organic layer was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and spin-dried to give crude product 27-1 (0.3 g, 39.7%). ¹ H NMR(600MHz,DMSO-d ₆ )δ12.0(s,1H)7.72(s,1H),3.73(m,2H),3.38(t,2H),2.37–2.31(m,2H),2.04(m,2H),1.92(m,2H).

(2) Synthesis of Compound 27

Intermediate 27-1 (0.10 g,0.34 mmol) was dissolved in anhydrous DCM (5 ml), the reaction was placed in ice-bath, 2-3 drops of DMF were added dropwise, oxalyl chloride (2M, 0.2 ml) was added under nitrogen protection, and after 2h reaction in ice-bath, the reaction was spun-dry to give intermediate 27-2.

1-6 (0.061 g,0.41 mmol) was placed in a reaction flask of intermediate 27-2 and placed in ice-bath, 5ml of anhydrous pyridine was added, after 2h TThe LC reaction was complete, extracted with EA and water, the organic layer was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and spin-dried to give product 27 (0.03 g, 17.3%) by column chromatography (DCM/meoh=20/1). ¹ H NMR(400MHz,DMSO-d ₆ )δ10.96(s,1H),8.63(s,1H),8.07(s,1H),7.84(s,1H),7.76(dd,1H),7.58(d,1H),4.35(m,2H),3.73(m,2H),3.53(t,2H),2.33(m,2H),2.02-2.10(m,2H),1.85-1.90(m,2H).

Test examples biological Activity test

1. The detection method comprises the following steps: whole-cell manual patch clamp technique to detect effect of compound on voltage-gated Nav1.8 channel current 2. Preparation and analysis of detection compound

Negative control: electrophysiological extracellular fluid containing 0.5% DMSO

Positive control: VX-150 as positive control drug

Test compound: a mass of the compound was weighed and dissolved in DMSO to prepare a 20mM DMSO stock solution. On the day of testing, 20mM of compound stock solution is diluted by extracellular fluid gradient to the final concentration to be detected, so that the DMSO content in the test drug solution is not more than 0.5%, and the concentration of DMSO has no influence on the detected Nav1.8 channel current. For example, 100nM and 1. Mu.M compound solutions were prepared and the gradient diluted as follows: firstly, sucking 5 mu L of DMSO mother liquor, adding the DMSO mother liquor into 10mL of extracellular fluid, and uniformly dissolving to obtain 10 mu M compound solution; then 1mL of 10 mu M compound is absorbed and added into 9mL of extracellular fluid, and the solution is uniformly dissolved to obtain 1 mu M compound solution; then, 1mL of the 1. Mu.M compound was taken up, added to 9mL of extracellular fluid, and dissolved uniformly to obtain a 100nM compound solution.

3. Cell culture

(1) Nav1.8 cell line: HEK293 (Flp-In T-Rex-293) cells stably expressing human Nav1.8 sodium channels, the coding gene information is as follows: NM-001293306.2.

(2) Culture and passaging conditions and methods: cell lines were cultured at 37℃with 5% CO ₂ Is placed in a constant temperature incubator. Nav1.8 stable transformants were cultured in complete medium containing 10% tetracycline-free fetal bovine serum (HyClone) and 100. Mu.g/mL Hygromycin B in DMEM (Gibco). Experiment beforeThe cells were digested and passaged when they grew to a density of about 90%, the medium was first aspirated, the cells were washed with Phosphate Buffer (PBS) pre-heated at 37℃and after removal of the PBS buffer, the cells were transferred to a centrifuge tube after digestion with pancreatin, centrifuged at 800rpm for 3 minutes, the supernatant was discarded, the cells were resuspended in complete medium containing 1. Mu.g/mL Doxcycline, passaged to 6-well plates, after 20 hours of induction culture, isolated and passaged to coverslips coated with polylysine for continued culture for 1-2 hours, and used for electrophysiological recording experiments.

4. Electrophysiological experiments

(1) Nav1.8 sodium channel current was recorded at room temperature (23-25 ℃) using whole cell voltage clamp technique.

(2) The whole-cell voltage clamp recording experiment adopts an Axon patch 700B patch clamp amplifier (Molecular Devices company), a digital-to-analog converter is Digidata 1440A (Molecular Devices company), a glass microelectrode is formed by drawing a glass electrode blank (World Precision Instrunents company) through a drawing instrument (P97, sutter company), the tip resistance after filling the electrode internal liquid is about 1.5-2.5MΩ, and the glass microelectrode can be connected to the patch clamp amplifier after being inserted into an amplifier probe. The clamp voltage and data were recorded and controlled by pClamp 10 software (Molecular Devices company) via a computer with a sampling frequency of 20kHz and a filtering frequency of 2kHz.

(3) Extracellular and intracellular fluids for electrophysiological experiments:

extracellular fluid formulation: 140mM NaCl,3mM KCl,1mM CaCl ₂ ,1mM MgCl ₂ 10mM HEPES and 20mM glucose, pH was adjusted to 7.3 with NaOH.

Intracellular fluid formulation: 140mM CsF,10mM NaCl,10mM HEPES,1.1mM EGTA and 20mM glucose, and pH was adjusted to 7.3 with CsOH.

Abbreviation notes: HEPES:4- (2-hydroxyethyl) piperazine-1-ethanesulfonic acid, N- (2-hydroxyethyl) piperazine-N' - (2-ethanesulfonic acid); EGTA: ethylene glycol bis (2-aminoethylether) tetraacetic acid; all drugs were purchased from Sigma.

(4) Electrophysiological stimulation protocol: after whole cell recordings were obtained, the electrophysiological recordings were started (high impedance gΩ sealing conditions were reached) after waiting for 4-5 minutes at-80 mV clamping voltage until the electrode inner liquid equilibrated with the cell inner liquid. Current stimulation and compound activity detection protocol: cells were clamped at-80 mV and stimulated with depolarizing voltages of 20ms, +10mV for repolarization to-80 mV at a stimulation frequency of 0.5Hz. After the Nav1.8 sodium channel current was determined to stabilize (about 1 minute) the dosing process was started until the cell current was no longer changing (compound inhibition reached steady state). At least 3 cells (n.gtoreq.3) were tested per concentration of compound. A single concentration of 100nM VX-150 was administered as a positive control after all compounds tested.

5. Data analysis

Data collection analysis the data were expressed as mean.+ -. Standard error (mean.+ -. SEM) using pClamp10 (Molecular Devices) GraphPad Prism 5 (GraphPad Software) and Excel (Microsoft corporation) software. The effect of a compound on current was calculated using the following formula:

inhibition ratio (%) = [ 1-magnitude of post-dosing current (I _Drug ) Magnitude of current before dosing (I _Control )]×100。

Dose response curves were fitted using Hill equation: y=bottom+ (Top-Bottom)/(1+10 (LogIC) ₅₀ -X) X k), wherein Bottom and Top represent minimum and maximum values of inhibition, respectively, X represents a logarithmic value of compound concentration, Y represents I _Drug /I _Control Numerical value, IC ₅₀ Represents the dose of drug that produces half the inhibitory effect, k represents the Hill coefficient.

The results are shown in tables 1 and 2.

TABLE 1 Activity of example Compounds against Navl.8 channel IC ₅₀ Value of

TABLE 2 percent blocking Activity of example compounds against Nav1.8 channels

The above embodiments are only for illustrating the technical solution of the present invention, and are not limited thereto. Although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: modifications may be made to the technical solutions described in the foregoing embodiments, or equivalents may be substituted for some or all of the technical features thereof, without departing from the spirit and scope of the present invention as defined in the claims; and such modifications or substitutions are intended to be within the scope of the present invention as defined by the claims.

Claims (10)

1. A compound of formula I, isomers, racemates, prodrugs or pharmaceutically acceptable salts thereof,

wherein:

x is selected from N or CH; y is selected from N or CR ₃ ；

V and G are each independently selected from N or CH, Q and T are each independently selected from N or C;

A. w and Z are each independently selected from O, S, N, carbonyl, sulfoxide, sulfone, -NR _a -、-CR _b -、-NR _a -CO-、-CR _b ＝N-、-CR _b -NR _a -, and at least one of A, W and Z contains nitrogen;

R _a independently at each occurrence selected from hydrogen, amino, hydroxy, C1-C6 alkyl, halo C1-C6 alkyl, C1-C6 alkoxy, halo C1-C6 alkoxy, C1-C6 alkylamino, C3-C8 cycloalkyl, 3-8 membered heterocyclyl containing 1 to 4 heteroatoms selected from N, O, S, C6-C12 aryl, or 5-10 membered heteroaryl containing 1 to 4 heteroatoms selected from N, O, S;

R _b independently at each occurrence selected from hydrogen, halogen, nitro, amino, cyano, hydroxy, C1-C6 alkyl, halo C1-C6 alkyl, C1-C6 alkoxy, halo C1-C6 alkoxy, C1-C6 alkylamino, C3-C8 cycloalkyl, 3-8 membered heterocyclyl containing 1 to 4 heteroatoms selected from N, O, S, C6-C12 aryl, or 5-10 membered heteroaryl containing 1 to 4 heteroatoms selected from N, O, S;

n is selected from 0, 1 or 2; in particular 2;

R ₁ selected from hydrogen, hydroxy, halogen, cyano, nitro or amino;

R ₂ Selected from halogen, hydroxy, cyano, nitro or halogenated C1-C6 alkyl;

R ₃ selected from halogen, hydroxy, cyano, amino, C1-C6 alkyl, halo C1-C6 alkyl, C2-C6 alkenyl, halo C2-C6 alkenyl, C2-C6 alkenyloxy, halo C2-C6 alkenyloxy, C2-C6 alkynyl, halo C1-C6 alkynyl, C2-C6 alkynyloxy, halo C1-C6 alkynyloxy, C1-C6 alkoxy, halo C1-C6 alkoxy, C1-C6 alkylamino, halo C1-C6 alkylamino, C3-C6 cycloalkylamino, C1-C6 alkylamino, C3-C6 cycloalkyl, C3-C6 cycloalkoxy or a 3-8 membered heterocyclyl containing 1 to 4 heteroatoms selected from N, O, S;

R ₆ and R is ₇ Each independently selected from hydrogen, fluorine, chlorine, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy;

R ₅ 、R ₈ and R is ₉ Each independently selected from hydrogen, fluorine, chlorine, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl;

represents a single bond or a double bond.

2. The compound, isomer, racemate, prodrug or pharmaceutically acceptable salt thereof according to claim 1, wherein,

R ₁ selected from hydrogen, fluorine, chlorine, bromine, amino or hydroxyl;

R ₂ selected from chlorine, bromine, iodine or trifluoromethyl;

R ₃ selected from fluorine, chlorine, bromine, amino, hydroxyl, methyl, cyclopropyl, methoxy, trifluoromethyl, trifluoromethoxy;

R ₆ And R is ₇ Each independently selected from hydrogen, fluorine, chlorine, methyl, ethyl or isopropyl;

R ₅ 、R ₈ and R is ₉ Each independently selected from hydrogen, methyl, ethyl, isopropyl, or cyclopropyl.

3. The compound, isomer, racemate, prodrug or pharmaceutically acceptable salt thereof according to claim 1, wherein,

the compound of formula I is selected from the following compounds of formula II:

wherein R is ₃ The definitions of A, Z, W, Q, T, V and G correspond to the claims.

4. The compound, isomer, racemate, prodrug or pharmaceutically acceptable salt thereof according to any of claim 1 to 3,

is one selected from the following groups:

5. the compound, isomer, racemate, prodrug or pharmaceutically acceptable salt thereof according to claim 1, wherein,

the compound of formula I is selected from the following compounds:

6. a process for the preparation of a compound as claimed in any one of claims 1 to 5 comprising the steps of:

carrying out acylation reaction on the formula III and the formula IV in the presence of alkali to obtain a compound of a formula I;

preferably, the base is selected from pyridine, sodium carbonate, sodium bicarbonate.

7. A pharmaceutical composition comprising a compound according to any one of claims 1-5, an isomer, a racemate, a prodrug or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable adjuvant.

8. Use of a compound according to any one of claims 1 to 5, an isomer, a racemate, a prodrug or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 7 for the preparation of a nav1.8 inhibitor.

9. Use of a compound according to any one of claims 1-5, an isomer, a racemate, a prodrug or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 7 in the manufacture of a medicament for the treatment, prevention or control of a disease or condition associated with the nav1.8 channel.

10. The use according to claim 9, wherein,

the Nav1.8 channel related diseases or conditions include nociceptive pain, inflammatory pain, neuropathic pain, functional pain, muscle or bone injury related pain, pelvic pain, abdominal pain, chest pain, lumbosacral neuralgia, preoperative pain, intraoperative pain, postoperative pain, acute or chronic pain, migraine, trigeminal neuralgia, pancreatitis, renal colic, cancer pain, pain resulting from chemical or drug therapy, diabetic neuralgia, postherpetic neuralgia, back pain, phantom limb pain, sciatica, small fiber neuralgia, erythromelalgia, arthritis, pruritus, acute or chronic pruritus, asthma, multiple sclerosis, arrhythmia, atrial fibrillation, heart failure, brugada syndrome, kidney stones, epilepsy, convulsions.