Amide compounds, processes for their preparation and uses thereof

Abstract

This invention provides an amide compound of Formula I, its preparation method, and its uses. The amide compound of this invention possesses selective inhibitory activity against Nav1.8 and can be used as a selective inhibitor of Nav1.8, exhibiting better activity, higher selectivity, and fewer side effects. It can be used to treat, prevent, or control diseases related to Nav1.8 channel involvement or dysfunction, and has significant clinical application value.

Description

Technical Field

[0001] This invention relates to the field of inhibitor synthesis technology, and in particular to a class of amide compounds, their preparation methods, and their use in treating diseases related to pain associated with the Nav1.8 target. Background Technology

[0002] Pain serves as a protective mechanism, alerting and protecting tissues from further damage. Pain is primarily generated when nociceptors convert received stimuli into nerve impulses (action potentials), which are then transmitted to the central nervous system via afferent nerve fibers, causing the sensation of pain. The generation and conduction of action potentials in neurons depend on voltage-gated sodium channels (Nav) on the cell membrane.

[0003] Voltage-gated sodium channels mediate the selective transmembrane flow of sodium ions and play a crucial role in the initiation, conduction, and transmission of action potentials in excitable cells such as neurons (Catterall et al., Pharmacol Rev. 2005, 57(4):397-409.). Nav channels are important drug targets, and Nav channel inhibitors are used to treat diseases such as pain, arrhythmia, epilepsy, anesthesia, and pruritus (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, nine channel subtypes, Nav1.1 to Nav1.9, have been identified in the mammalian genome. Based on amino acid sequence homology, the similarity of Nav channel proteins ranges from 45% to 87%. Based on their different sensitivities to tetrodotoxin (TTX), they can be divided into TTX-tolerant channels (TTX-R) and TTX-sensitive channels (TTX-S). Among them, Nav1.5, Nav1.8 and Nav1.9 channels belong to the TTX-R type sodium channels, while other subtypes belong to the TTX-S subtype (Catterall et al., Pharmacol Rev. 2005, 57(4):397-409.).

[0004] Voltage-gated Nav1.8 channel subtypes (TTX-R type) are mainly distributed in the peripheral nervous system, such as expressing Nav1.8 channels in 75% of dorsal root neurons. Because Nav1.8 channels have high activation and inactivation voltages, they are a major component of the rising limb of action potentials (other Nav channel subtypes are in a nonfunctional inactivated state) (Goodwin et al., Nat RevNeurosci. 2021, 22(5):263-274.). Due to the slow inactivation and rapid reactivation characteristics of Nav1.8 channels, they participate in the physiological and pathological processes of membrane potential depolarization and high-frequency neuronal discharge, such as pain (Alsaloum et al., Nat RevNeurol. 2020, 16(12):689-705.). Human genetic studies have shown that mutations in the Nav1.8 gene lead to small fiber neuralgia and erythropathic pain (Faber et al., Proc Natl Acad Sci US A. 2012, 109(47):19444-9; Kaluza et al., Pflugers Arch. 2018, 470(12):1787-1801.). In rodents, gene knockout or knockdown of the Nav1.8 channel gene can alleviate various types of inflammatory pain and neuralgia; while administration of Nav1.8 channel inhibitors such as A-803467 can effectively alleviate pain responses (Jarvis et al., Proc Natl Acad Sci US A. 2007, 104(20):8520-5.). Diabetic neuropathy is one of the most common neuropathic pain disorders, affecting approximately 60%–70% of diabetic patients, with over 70% of patients not receiving effective treatment (Jensen et al., Brain. 2021, 144(6): 1632–1645.). Pyrone aldehyde directly enhances Nav1.8 channel function in diabetic neuropathy patients, and gene knockout or reduction of Nav1.8 channels can effectively alleviate neuropathy (Bierhaus et al., Nat Med. 2012, 18(6): 926–33.). In a rat model of STZ-induced diabetic neuropathy, intraperitoneal or plantar administration of the Nav1.8 channel inhibitor A-803467 dose-dependently alleviated pain behavior responses in animals (Mert et al., J Am Assoc Lab Anim Sci. 2012, 51(5): 579–85.).

[0005] Besides pain, the Nav1.8 channel is also associated with multiple sclerosis, arrhythmia, cough, pruritus, and epilepsy. Multiple sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system, and its exact pathogenesis remains to be elucidated. Normal Purkinje fibers in the cerebellum do not express the Nav1.8 channel, but the expression of Nav1.8 in the cerebellum of patients with multiple sclerosis is upregulated, and the expression level of the channel increases in a disease-dependent manner as the disease progresses. The single nucleotide polymorphism (SNP) of the Nav1.8 encoding gene is also related to the severity of MS (Craner et al., J Neuropathol Exp Neurol. 2003, 62(9):968-75; Roostaei et al., Neurology. 2016, 86(5):410-7.). Mouse transgenic mice with Nav1.8 knock-in (overexpression) in the cerebellum Purkinje fibers (L7-1.8TG) exhibited multiple sclerosis (MS) behavior. Administration of the 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):e0119067.). Osteoarthritis is a degenerative joint disease characterized by cartilage wear and pain. Phosphorylated cAMP response element-binding protein (CREB) directly binds to the promoter of the Nav1.8 encoding gene, promoting Nav1.8 protein transcription and upregulating Nav1.8 channel expression levels (Zhu et al., Elife. 2020, 9:e57656.). In the cardiovascular system, the Nav1.8 channel has been shown to be expressed in cardiac nerves such as Purkinje fibers, and some studies also suggest that Nav1.8 is expressed in cardiomyocytes (Verkerk et al., Circ Res. 2012, 111(3):333-43.). Human genetic studies have found that mutations in the Nav1.8 gene are associated with Brugada syndrome (Hu et al., J Am Coll Cardiol. 2014, 64(1):66-79.). Inhibition of the Nav1.8 channel can improve cardiac remodeling, and the Nav1.8 channel is considered a potential therapeutic target for cardiovascular diseases such as arrhythmias, atrial fibrillation, and heart failure (Dybkova et al., Cardiovasc Res. 2018, 114(13):1728-1737.). Nav1.8 channels are expressed in the cough-related vagus plexus. The phosphorylation level and expression of Nav1.8 are increased during pathological cough, and it is involved in the cough reflex (Muroi et al., Lung. 2014, 192(1): 15-20.).In mammalian itch sensation, itch-inducing factors such as histamine released by lymphocytes and mast cells can activate the Nav1.8 channel. Knockout of Nav1.8 in mice can effectively alleviate histamine- and endothelin-induced pruritus (Riol-Blanco et al., 2014, 510(7503):157-61.). In addition, congenital mutations in human Nav1.8 have been reported to cause epilepsy and seizures (Kambouris et al., Ann Clin Transl Neurol. 2016, 4(1):26-35.).

[0006] Currently, the only selective Nav1.8 inhibitor is VX-150 from Vertex Pharmaceuticals, which has completed Phase II clinical trials in patients with osteoarthritis, acute pain, and pain caused by small fiber neuropathy, with positive results. In China, Hengrui's HRS-4800 is the only selective Nav1.8 inhibitor currently in clinical trials, undergoing Phase I trials. Several other selective inhibitors are in the preclinical development stage. Therefore, developing Nav1.8 inhibitors with better activity, higher selectivity, and fewer side effects has significant clinical application and innovative drug value.

[0007] In view of this, the present invention is hereby proposed. Summary of the Invention

[0008] One of the objectives of this invention is to provide an amide compound with selective Nav1.8 inhibitory activity.

[0009] A second objective of this invention is to provide a method for preparing amide compounds.

[0010] A third objective of this invention is to provide a pharmaceutical composition comprising the aforementioned amide compound.

[0011] A fourth objective of this invention is to provide the use of the aforementioned amide compound or pharmaceutical composition in the preparation of Nav1.8 inhibitors or in the preparation of medicaments for the treatment, prevention or control of diseases or symptoms related to the Nav1.8 channel.

[0012] In order to achieve the above-mentioned objectives of the present invention, the following technical solution is adopted:

[0013] This invention provides, in one aspect, a compound of formula I, its isomers, racemates, prodrugs, or pharmaceutically acceptable salts thereof.

[0014]

[0015] in:

[0016] X is selected from N or CH; Y is selected from N or CR3;

[0017] V and G are each independently selected from N or CH, and Q and T are each independently selected from N or C;

[0018] A, W, and Z are each independently selected from O, S, N, carbonyl, sulfoxide, sulfone, and -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;

[0019] R a Each time it appears, it is independently selected from hydrogen, amino, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C1-C6 alkylamino, C3-C8 cycloalkyl, 3-8 membered heterocyclic group containing 1 to 4 heteroatoms selected from N, O, S, C6-C12 aryl or 5-10 membered heteroaryl group containing 1 to 4 heteroatoms selected from N, O, S;

[0020] R b Each time it appears, it is independently selected from hydrogen, halogen, nitro, amino, cyano, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C1-C6 alkylamino, C3-C8 cycloalkyl, 3-8 membered heterocyclic group containing 1 to 4 heteroatoms selected from N, O, S, C6-C12 aryl or 5-10 membered heteroaryl group containing 1 to 4 heteroatoms selected from N, O, S;

[0021] n is selected from 0, 1, or 2; especially 2;

[0022] R1 is selected from hydrogen, hydroxyl, halogen, cyano, nitro or amino; preferably hydrogen, fluorine, chlorine, bromine, amino or hydroxyl.

[0023] R2 is selected from halogens, hydroxyl groups, cyano groups, nitro groups, or halogenated C1-C6 alkyl groups; preferably chlorine, bromine, iodine, or trifluoromethyl.

[0024] R3 is selected from halogens, hydroxyl groups, cyano groups, amino groups, C1-C6 alkyl groups, halogenated C1-C6 alkyl groups, C2-C6 alkenyl groups, halogenated C2-C6 alkenyl groups, C2-C6 alkenoxy groups, halogenated C2-C6 alkenoxy groups, C2-C6 alkynyl groups, halogenated C1-C6 alkynoxy groups, C1-C6 alkoxy groups, halogenated C1-C6 alkoxy groups, C1-C6 alkylamino groups, halogenated C1-C6 alkylamino groups, C3-C6 cycloalkylamino groups, C1-C6 alkoxyamino groups, C3-C6 cycloalkyl groups, C3-C6 cycloalkoxy groups, or 3-8 membered heterocyclic groups containing 1 to 4 heteroatoms selected from N, O, and S; preferably fluorine, chlorine, bromine, amino, hydroxyl, methyl, cyclopropyl, methoxy, trifluoromethyl, or trifluoromethoxy.

[0025] R6 and R7 are each independently selected from hydrogen, fluorine, chlorine, C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy, and C1-C6 alkoxy; preferably, each is independently selected from hydrogen, fluorine, chlorine, methyl, ethyl, or isopropyl.

[0026] R5, R8 and R9 are each independently selected from hydrogen, fluorine, chlorine, halogenated C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkoxy, C3-C8 cycloalkyl; preferably, each is independently hydrogen, methyl, ethyl, isopropyl or cyclopropyl.

[0027] Indicates a single bond or a double bond.

[0028] In some embodiments, the compound of formula I is selected from the compounds of formula II:

[0029]

[0030] The definitions of R3, A, Z, W, Q, T, V, and G are the same as those described above.

[0031] In some preferred embodiments, It is selected from one of the following groups:

[0032]

[0033] In some preferred embodiments, the compound of formula I is selected from the following compounds:

[0034]

[0035]

[0036]

[0037]

[0038] The terms used in this invention are defined as follows:

[0039] "Halogen" can be fluorine, chlorine, bromine or iodine.

[0040] "C1-C6" alkyl refers to a chain alkyl group having 1-6 carbon atoms; specific examples may include methyl, ethyl, propyl, n-propyl, isopropyl, butyl, n-butyl, isobutyl, tert-butyl, 1-methyl-butyl, 1-ethyl-butyl, pentyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, hexyl, n-hexyl, 1-methylpentyl, 2-methylpentyl, 4-methyl-2-pentyl, 3,3-dimethylbutyl, 2-ethylbutyl, and similar groups; "halogenated alkyl" refers to a group obtained by substituting at least one hydrogen atom of the alkyl group as described above with a halogen; specific examples include trifluoromethyl, etc.

[0041] "C2-C6 alkenyl" refers to a straight-chain or branched group containing 2-6 carbon atoms and at least one carbon-carbon double bond; specific examples may include vinyl, 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)-3-hexenyl, (E)-1,3-hexadienyl, 4-methyl-3-pentenyl, or norbornene.

[0042] "C2-C6 ynyl" refers to a straight-chain or branched group containing 2-6 carbon atoms and at least one carbon-carbon double bond; specific examples may include ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, 1-hexynyl, 2-hexynyl, and 3-hexynyl.

[0043] "C1-C6 alkoxy" refers to the RO- group, where R is a C1-C6 alkyl group as described above. Specific examples of alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, sec-butoxy, n-pentoxy, isopentoxy, neopentoxy, n-hexyloxy, isohexyloxy, 3-methylpentoxy, 3,3-dimethylbutoxy, 2-ethylbutoxy, etc. "Haloalkoxy" refers to a group obtained by substituting at least one hydrogen atom of an alkoxy group as described above with a halogen; specific examples include trifluoromethoxy, etc.

[0044] "C2-C6 olefinic group" refers to the RO- group, where R is a C2-C6 olefinic group as described above; specific examples of olefinic groups include ethylene oxy and propylene oxy groups.

[0045] "C2-C6 alkynyloxy" refers to the RO- group, where R is the C2-C6 alkynyl group as described above; specific examples of alkynyloxy include acetyloxy and propynyloxy.

[0046] "Amino" refers to -NH2.

[0047] "C1-C6 alkylamino" refers to a group obtained by substituting one or two hydrogens of -NH2 with a C1-C6 alkyl group as described above, and can be represented as R1R2N-, where R1 and R2 are each independently H or C1-C6 alkyl, and at most one of R1 and R2 is H. Specific examples of C1-C6 alkylamino groups include methylamino, dimethylamino, ethylamino, n-propylamino, isopropylamino, n-butylamino, isobutylamino, tert-butylamino, sec-butylamino, n-pentamino, isopentamino, neopentamino, n-hexylamino, isohexylamino, 3-methylpentamino, 3,3-dimethylbutylamino, 2-ethylbutylamino, etc.

[0048] "C1-C6 alkoxyamino" refers to a group obtained by replacing the two hydrogens of -NH2 with C1-C6 alkyl and oxygen as described above; specific examples of C1-C6 alkoxyamino include methoxyamine, dimethoxyamine, ethoxyamine, n-propoxyamine, and isopropoxyamine.

[0049] "C3-C8 cycloalkyl" refers to a fully saturated cyclic hydrocarbon compound group containing 3-8 carbon atoms, and specific examples include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0050] "C3-C6 cycloalkylamino" refers to a group obtained by substituting one or two hydrogens of -NH2 with a C3-C6 cycloalkyl group as described above, and can be represented as R1R2N-, where R1 and R2 are each independently H or C3-C6 cycloalkyl, and at most one of R1 and R2 is H. Specific examples of C3-C6 cycloalkylamino groups include cyclopropylamino, cyclobutanamino, cyclopentanamino, cyclohexylamino, etc.

[0051] "3-8 membered heterocyclic group" refers to a 3-8 membered non-aromatic cyclic alkyl group that contains at least one heteroatom selected from nitrogen, oxygen, and sulfur on the ring; specific examples include piperazine, piperidine, morpholine, etc.

[0052] "C6-C12 aryl" refers to a monocyclic or polycyclic aryl group having 6 to 12 carbon atoms; specific examples include phenyl and naphthyl.

[0053] "5-10-membered heteroaryl" refers to a 5-10-membered aromatic group containing at least one heteroatom selected from nitrogen, oxygen, and sulfur on the ring; specific examples 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, indole, isoindole, etc.

[0054] "Pharmaceutically acceptable salts" include salts formed by the compound of Formula I with an acid or base; the acid includes inorganic acids and organic acids; preferably, the inorganic acid includes hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, and carbonic acid; preferably, the organic acid includes 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, and dihydroxynaphthyl acid; the base includes hydroxides, carbonates, and bicarbonates of sodium, potassium, calcium, aluminum, lithium, and ammonium.

[0055] The compounds and their pharmaceutically acceptable salts involved in this application may have isomers or racemates, such as optical isomers (including diastereomers and enantiomers), transisomers, geometric isomers (cis-trans isomers), conformational isomers, tautomers, and mixtures thereof, but are not limited thereto. These isomers are also included within the scope defined by the claims of this invention.

[0056] Another aspect of the present invention provides a method for preparing the compound of formula I above, which is achieved through the following reaction route:

[0057]

[0058] Formula I is obtained by acylation of Formulas III and IV in the presence of a base.

[0059] Preferably, the base is selected from pyridine, sodium carbonate, and sodium bicarbonate.

[0060] In another aspect, the present invention provides a pharmaceutical composition comprising one or more selected from compounds of formula I and their isomers, racemates, pharmaceutically acceptable salts and prodrugs, and optionally pharmaceutically acceptable excipients.

[0061] In another aspect, the present invention provides the use of a compound of formula I or an isomer thereof, a racemate, a pharmaceutically acceptable salt thereof, or a prodrug in the preparation of a Nav1.8 inhibitor or in the preparation of a medicament for the treatment, prevention or control of diseases or symptoms associated with the Nav1.8 channel.

[0062] In another aspect, the present invention provides a method for treating, preventing, or controlling diseases or symptoms associated with Nav1.8, the method comprising administering to a subject in need one or more selected from compounds of formula I, isomers thereof, racemates, pharmaceutically acceptable salts and prodrugs, or combinations thereof.

[0063] Diseases or symptoms associated with the Nav1.8 channel include, but are not limited to, nociceptive pain, inflammatory pain, neuropathic pain, functional pain, muscle or skeletal 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 caused by chemotherapy or drug therapy, diabetic neuropathy, 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, and seizures.

[0064] The present invention has the following beneficial effects:

[0065] The amide compounds of this invention have selective inhibitory activity against Nav1.8 and can be used as selective inhibitors of Nav1.8. They have better activity, higher selectivity, and fewer side effects, and can be used to treat, prevent or control diseases related to Nav1.8 channel involvement or dysfunction, thus having important clinical application value.

[0066] The present invention has been described in detail above; however, the above embodiments are merely illustrative in nature and are not intended to limit the invention. Furthermore, this document is not limited to the foregoing prior art or the invention itself, or to any theory described in the following embodiments. Detailed Implementation

[0067] The present invention will be further described below with reference to the embodiments. It should be noted that the following embodiments are provided for illustrative purposes only and do not constitute a limitation on the scope of protection of the present invention.

[0068] Unless otherwise specified, the raw materials, reagents, and methods used in the embodiments are all conventional raw materials, reagents, and methods in the art.

[0069] The Chinese names of reagents represented by chemical formulas or English letter abbreviations are as follows: ℃ represents degrees Celsius; g represents grams; s represents singlet, d represents doublet, t represents triplet, m represents multiplet; min represents minutes; ml represents milliliters; mmol represents millimoles; h represents hours; TLC represents thin-layer chromatography.

[0070] In the following examples, the proton NMR spectra were recorded using a Bruker AMX-400 or AMX-600 NMR spectrometer, and the chemical shift δ was expressed in ppm.

[0071] Unless otherwise specified, all reaction solvents were purified using standard methods.

[0072] Thin-layer chromatography used GF254 high-efficiency plates, which were produced by Yantai Chemical Research Institute.

[0073] Unless otherwise specified, all solvents are analytical grade reagents, and all reagents used were purchased from Sinopharm Chemical Reagent Co., Ltd.

[0074] The colorimetric method employed included 2,4-dinitrophenylhydrazine, iodine, and ultraviolet fluorescence.

[0075] The removal of organic solvents under reduced pressure is carried out in a rotary evaporator.

[0076] Example 1: Synthesis of Compound 1

[0077] [Reaction Route 1]

[0078]

[0079] (1) Synthesis of intermediates 1-2

[0080] Compound 1-1 (12.2 g, 57.2 mmol) was dissolved in anhydrous dichloromethane (300 ml) and placed in an ice bath. Under nitrogen protection, bis(2-methoxyethyl)aminosulfur trifluoride (BAST) was slowly added dropwise. After the addition was complete, the mixture was moved to room temperature and reacted for 24 h. TLC showed that the reaction was almost complete. The reaction was quenched by slowly adding saturated sodium bicarbonate solution under ice bath conditions, and a large amount of gas was generated. The mixture was extracted with dichloromethane (DCM) and water. The organic layer was dried with anhydrous sodium sulfate and separated by column chromatography (petroleum ether (PE) / ethyl acetate (EA) = 5 / 1) to give intermediate 1-2 (10 g, 74.3%). 1 H NMR (400MHz, CDCl3) 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).

[0081] (2) Synthesis of intermediates 1-3

[0082] Intermediate 1-2 (10 g, 42.5 mmol) was placed in a 250 ml round-bottom flask, and 4N dioxane hydrochloride solution (42.5 ml, 170 mmol) was added under ice bath conditions. The reaction was carried out at room temperature for about 4 h. The reaction was confirmed to be complete by TLC. The reaction solution was directly evaporated to dryness to obtain 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).

[0083] [Reaction Route 2]

[0084]

[0085] (3) Synthesis of intermediates 1-5

[0086] Compounds 1-4 (0.9 g, 6.76 mmol) were dissolved in 10 ml of concentrated sulfuric acid and transferred to an ice bath. Potassium nitrate (0.68 g, 6.76 mmol) was added and the mixture was reacted for 3 h in an ice bath. TLC showed that the reaction was almost complete. The reaction solution was slowly added to ice water, and a large amount of white solid precipitated out. The solid was filtered to obtain a filter cake, which was then dried to obtain intermediate 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).

[0087] (4) Synthesis of intermediates 1-6

[0088] Intermediate 1-5 (1.1 g, 6.17 mmol) was dissolved in 100 ml of methanol, and palladium on carbon (0.22 g) with a water content of 60% was added. After purging with hydrogen three times, the mixture was heated to 40 °C and reacted for another 7 h. The reaction was confirmed to be complete by TLC. The reaction solution was filtered and the filtrate was evaporated to dryness to obtain 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).

[0089] [Reaction Route 3]

[0090]

[0091] (5) Synthesis of intermediates 1-8

[0092] Intermediate 1-3 (2.38 g, 12.8 mmol) was dissolved in 40 mL of N,N-dimethylformamide (DMF), potassium carbonate (5.5 g, 38.5 mmol) was added, followed by compound 1-7 (2.0 g, 11.7 mmol). The mixture was heated to 120 °C and reacted overnight. TLC analysis showed that the reaction was essentially complete. The mixture was extracted with EA and water, the organic layer was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and then subjected to column chromatography (PE / EA = 5 / 1) to obtain intermediate 1-8 (2.0 g, 54.9%). 1H NMR(400MHz, CDCl3) 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).

[0093] (6) Synthesis of intermediates 1-9

[0094] Intermediate 1-8 (2.0 g, 7.04 mmol) was dissolved in DMAC (N,N-dimethylacetamide) (40 ml), and NCS (N-chlorosuccinimide) (1.8 g, 14.1 mmol) was added. The mixture was heated to 100 °C, and the reaction was checked for completeness after 1 h. The mixture was extracted with EA and water, the organic layer was washed with saturated sodium chloride, dried with anhydrous sodium sulfate, and subjected to column chromatography (PE / EA = 10 / 1) to obtain intermediate 1-9 (1.5 g, 66.8%). 1 H NMR (400MHz, CDCl3) 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).

[0095] (7) Synthesis of intermediates 1-10

[0096] Intermediate 1-9 (1.5 g, 4.7 mmol) was dissolved in methanol (40 ml), lithium hydroxide (2.0 g, 47 mmol) was added, and 8 ml of water was added. The mixture was heated to 50 °C and reacted for 4 h. TLC showed that the reaction was almost complete. The reaction solution was evaporated to dryness, and the pH of the reaction solution was adjusted to about 3-4 with 2N hydrochloric acid. A large amount of white solid precipitated out. The mixture was filtered to obtain intermediate 1-10 (1.15 g, 80.3%). 1 HNMR(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).

[0097] (8) Synthesis of intermediate 1-11

[0098] Intermediate 1-10 (0.05 g, 0.16 mmol) was dissolved in anhydrous DCM (5 ml). The reaction solution was placed in an ice bath and 2-3 drops of DMF were added as a catalyst. Oxaloyl chloride (2 M, 0.1 ml) was added under nitrogen protection. After reacting in an ice bath for 2 h, the reaction solution was evaporated to dryness to obtain intermediate 1-11.

[0099] (9) Synthesis of Compound 1

[0100] Intermediate 1-6 (0.026 g, 0.18 mmol) was added to intermediate 1-11 and placed in an ice bath. 3 mL of anhydrous pyridine was added dropwise. After 2 h, the reaction was detected by TLC to be complete. The mixture was extracted with EA and water. The organic layer was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and subjected 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).

[0101] Example 2: Synthesis of Compound 2

[0102] Following the method in step 9 of Example 1, intermediates 1-6 were replaced with 5-amino-2,3-dihydroisoindole-1-one (purchased from Bidex) to obtain 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).

[0103] Example 3: Synthesis of Compound 3

[0104] Following the method in step 9 of Example 1, intermediates 1-6 were replaced with 6-aminoindolone (purchased from Shaoyuan) to obtain compound 3 (0.020 g, 29.7%). 1H 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).

[0105] Example 4: Synthesis of Compound 4

[0106] [Reaction Route 4]

[0107]

[0108] (1) Synthesis of intermediate 4-2

[0109] Compound 4-1 (0.6 g, 3.4 mmol) was dissolved in 50 mL of methanol, and palladium on carbon (0.12 g) with a water content of 60% was added. After purging with hydrogen three times, the reaction was continued at room temperature for 7 h. The reaction was confirmed to be complete by TLC. The reaction solution was filtered, and the filtrate was evaporated to dryness to obtain 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).

[0110] (2) Synthesis of compound 4

[0111] Following the method in step 9 of Example 1, intermediates 1-6 were replaced with intermediates 4-2 to obtain 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).

[0112] Example 5: Synthesis of Compound 5

[0113] [Reaction Route 5]

[0114]

[0115] (1) Synthesis of intermediate 5-2

[0116] Compound 5-1 (1.9 g, 9.9 mmol) was dissolved in 10 mL of anhydrous ethanol, and hydrazine hydrate (85%) (1.5 g, 47.7 mmol) was added. The mixture was heated to reflux, and after 2 h, the reaction was detected by TLC to be complete. The mixture was filtered to obtain a filter cake, which was then dried to obtain crude product 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).

[0117] (2) Synthesis of intermediate 5-3

[0118] Intermediate 5-2 (1.05 g, 5.7 mmol) was dissolved in 60 mL of DCM, and triethylamine (0.7 g, 7.0 mmol) was added. Then, di-tert-butyl dicarbonate ((Boc)₂O) (1.5 g, 7.0 mmol) was added. The reaction was carried out at room temperature for 3 h. TLC showed that the reaction was basically complete. The mixture was extracted with DCM and water. The organic layer was washed with saturated sodium chloride (NaCl) and dried with anhydrous sodium sulfate (Na₂SO₄). The mixture was then subjected to rotary cyclohexane chromatography (DCM / MeOH = 20 / 1) to give 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).

[0119] (3) Synthesis of intermediate 5-4

[0120] Intermediate 5-3 (1.0 g, 4.0 mmol) was dissolved in 50 ml of methanol, and palladium on carbon (0.20 g) with a water content of 60% was added. After purging with hydrogen three times, the reaction was continued at room temperature for 7 h. The reaction was confirmed to be complete by TLC. The reaction solution was filtered and the filtrate was evaporated to dryness to obtain 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).

[0121] (4) Synthesis of compound 5

[0122] Referring to step 9 of Example 1, intermediates 1-6 were replaced with intermediate 5-4 to obtain intermediate 5-5. Intermediate 5-5 was dissolved in 5 ml of DCM, and 2 ml of trifluoroacetic acid (TFA) was added under ice bath conditions. The reaction was carried out at room temperature for 2 h. The reaction was detected by TLC to be complete. The mixture was evaporated to dryness, extracted with EA and water, and the organic layer was washed with saturated sodium bicarbonate and sodium chloride, respectively. After drying with anhydrous sodium sulfate, the mixture was subjected to rotary evaporation column chromatography (DCM / MeOH = 10 / 1) to give 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).

[0123] Example 6: Synthesis of Compound 6

[0124] [Reaction Route 6]

[0125]

[0126] (1) Synthesis of intermediate 6-2

[0127] Compound 6-1 (0.6 g, 2.6 mmol) was dissolved in 30 ml of methanol, and palladium on carbon (0.12 g) with a water content of 60% was added. After purging with hydrogen three times, the reaction was continued at room temperature for 7 h. The reaction was confirmed to be complete by TLC. The reaction solution was filtered, and the filtrate was evaporated to dryness to obtain intermediate 6-2 (0.4 g, 76.7%). 1 H NMR (400MHz, DMSO) δ7.59 (d, J = 8.5 Hz, 1H), 6.94 (d, J = 1.9 Hz, 1H), 6.89 (dd, J = 8.5, 2.0 Hz, 1H), 6.77 (s, 1H).

[0128] (2) Synthesis of intermediate 6-3

[0129] Intermediate 6-2 (0.4 g, 2.0 mmol) was dissolved in 5 ml of concentrated hydrochloric acid under ice bath conditions. Zinc powder (1.1 g, 16.2 mmol) was added in portions. After the addition was complete, the reaction was allowed to rise naturally to room temperature. After 2 h, the reaction was detected by TLC to be complete. The reaction solution was slowly poured into ice water to quench the reaction. The pH was adjusted to about 7 with 2N sodium hydroxide. Extraction was performed with EA and water. The organic layer was washed with saturated sodium chloride, dried with anhydrous sodium sulfate, and subjected to column chromatography (DCM / MeOH = 10 / 1) to obtain intermediate 6-3 (0.10 g, 27.1%). 1 HNMR (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).

[0130] (3) Synthesis of compound 6

[0131] Following step 9 of Example 1, intermediates 1-6 were replaced with intermediate 6-3 to obtain 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=4.5 Hz,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).

[0132] Example 7 Synthesis of Compound 7

[0133] [Reaction Route 7]

[0134]

[0135] (1) Synthesis of intermediate 7-2

[0136] Compound 7-1 (1.0 g, 5.6 mmol) was dissolved in 40 ml of methanol, and palladium on carbon (0.2 g) with a water content of 60% was added. After purging with hydrogen three times, the reaction was continued at room temperature for 7 h. The reaction was confirmed to be complete by TLC. The reaction solution was filtered, and the filtrate was evaporated to dryness to obtain intermediate 7-2 (0.7 g, 83.2%). 1H 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).

[0137] (2) Synthesis of compound 7

[0138] Referring to step 9 of Example 1, intermediates 1-6 were replaced with intermediates 7-2 to obtain 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=8.4H z,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).

[0139] Example 8: Synthesis of Compound 8

[0140] [Reaction Route 8]

[0141]

[0142] (1) Synthesis of intermediate 8-2

[0143] Compound 8-1 (2.0 g, 9.9 mmol) was dissolved in 20 mL of anhydrous ethanol, and hydrazine hydrate (85%) (2.92 g, 49.6 mmol) was added. The mixture was heated to reflux, and after 3 h, the reaction was detected by TLC to be complete. The mixture was filtered to obtain a filter cake, which was then dried to obtain 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).

[0144] (2) Synthesis of intermediate 8-3

[0145] The crude product 8-2 (2.0 g, 10.1 mmol) was dissolved in 30 ml of water, 3 ml of concentrated hydrochloric acid was added, and the mixture was heated to 90 °C and reacted overnight. The reaction was detected by TLC and was basically complete. The mixture was filtered to obtain a filter cake, which was then dried to obtain the crude product intermediate 8-3 (1.67 g, 91.9%). 1H 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).

[0146] (3) Synthesis of intermediate 8-4

[0147] Intermediate 8-3 (1.5 g, 8.4 mmol) was dissolved in 50 mL of DCM, and triethylamine (2.5 g, 25.1 mmol) was added, followed by (Boc)₂O (2 g, 9.2 mmol). The reaction was carried out at room temperature for 3 h. TLC showed that the reaction was basically complete. The mixture was extracted with DCM and water, the organic layer was washed with saturated NaCl, dried with anhydrous Na₂SO₄, and then subjected to rotary cyclohexane chromatography (DCM / MeOH = 20 / 1) to give 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).

[0148] (4) Synthesis of intermediate 8-5

[0149] Intermediate 8-4 (1.5 g, 5.4 mmol) was dissolved in 50 ml of methanol, and palladium on carbon (0.30 g) with a water content of 60% was added. After purging with hydrogen three times, the reaction was continued at room temperature for 7 h. The reaction was confirmed to be complete by TLC. The reaction solution was filtered and the filtrate was evaporated to dryness to obtain 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).

[0150] (5) Synthesis of compound 8

[0151] Referring to step 9 of Example 1, intermediate 1-6 was replaced with intermediate 8-5 to obtain intermediate 8-6 (0.091 g, 34.6%). Intermediate 8-6 was dissolved in DCM (5 ml), and 2 ml of TFA was added under ice bath conditions. The reaction was carried out at room temperature for 2 h. The reaction was detected by TLC to be complete. The mixture was evaporated to dryness, extracted with EA and water, and the organic layer was washed with saturated sodium bicarbonate and sodium chloride, respectively. The mixture was dried over anhydrous sodium sulfate and subjected to rotary evaporation column chromatography (DCM / MeOH = 10 / 1) to obtain compound 8 (0.05 g, 25.5%). 1H 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),7.2 6(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).

[0152] Example 9: Synthesis of Compound 9

[0153] Referring to step 9 of Example 1, intermediates 1-6 were replaced with intermediates 6-2 to obtain compound 9 (9 mg, 19.8%). 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).

[0154] Example 10: Synthesis of Compound 10

[0155] Referring to step 9 of Example 1, intermediates 1-6 were replaced with 4-aminophthalic acid hydrazide (purchased from Leyan) to obtain 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).

[0156] Example 11 Synthesis of Compound 11

[0157] [Reaction Route 9]

[0158]

[0159] (1) Synthesis of intermediate 11-2

[0160] Referring to step 9 of Example 1, intermediates 1-6 were replaced with compound 11-1 to obtain intermediate 11-2 (0.48 g, 69.1%). 1H 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).

[0161] (2) Synthesis of compound 11

[0162] Intermediate 11-2 (0.12 g, 0.28 mmol) was dissolved in ultradry DMF (3 ml), and potassium tert-butoxide (0.048 g, 0.426 mmol) and acetoxyxamic acid (0.032 g, 0.43 mmol) were added. After reacting at 50 °C for 4 h, the reaction was confirmed to be complete by TLC. The mixture was extracted with EA and water, the organic layer was washed with saturated sodium chloride and dried with anhydrous sodium sulfate, and then subjected to column chromatography (DCM / MeOH = 20 / 1) 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).

[0163] Example 12 Synthesis of Compound 12

[0164] [Reaction Route 10]

[0165]

[0166] Intermediate 11-2 (0.10 g, 0.28 mmol) was dissolved in ethanol (10 ml), and 0.3 ml of hydrazine hydrate solution was added. The mixture was heated to 80 °C and refluxed overnight. The reaction was confirmed to be complete by TLC the next day. The mixture was extracted with EA and water, the organic layer was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and then subjected to column chromatography (DCM / MeOH = 20 / 1) to give compound 12 (0.067 g, 65.1%). 1HNMR (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).

[0167] Example 13 Synthesis of Compound 13

[0168] [Reaction Route 11]

[0169]

[0170] (1) Synthesis of intermediate 13-1

[0171] 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), and potassium carbonate (7.0 g, 52 mmol) was added. The reaction solution was heated to 80 °C and reacted for 4 h. TLC showed that the reaction was basically complete. The reaction was quenched with water, and the pH was adjusted to about 4 with 2 M hydrochloric acid. Extraction was performed with EA and water. The organic layer was washed with saturated sodium chloride and dried with anhydrous sodium sulfate. The residue was then evaporated and separated by column chromatography (DCM / MeOH = 20 / 1) to obtain 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).

[0172] (2) Synthesis of intermediate 13-2

[0173] Compound 13-1 (0.5 g, 1.72 mmol) was dissolved in anhydrous dichloromethane (40 mL) and placed in an ice bath. Oxaloyl chloride (0.18 mL, 2.1 mmol) was added dropwise under nitrogen protection, along with a catalytic amount of DMF. The reaction was carried out under ice bath conditions for about 2 h. The reaction solution was directly evaporated to dryness to obtain the crude product, which was then placed in a reaction flask. 5-Amino-2-fluorobenzonitrile was added to this reaction flask, and anhydrous pyridine (5 mL) was added dropwise under nitrogen protection and an ice bath conditions. After 2 h, TLC showed that the reaction was almost complete. The mixture was extracted with EA and water. The organic layer was washed with saturated sodium chloride and dried with anhydrous sodium sulfate. The solution was evaporated to dryness and then subjected to column chromatography (DCM / MeOH = 50 / 1) to obtain 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).

[0174] (3) Synthesis of intermediate 13-3

[0175] 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). After purging with nitrogen 3-4 times, Pd(dppf)₂Cl₂ (0.07 g, 0.098 mmol) was added. The reaction solution was heated to 100 °C and reacted overnight. TLC showed that the reaction was basically complete. Extraction was performed with EA and water. The organic layer was washed with saturated sodium chloride and dried with anhydrous sodium sulfate. Column chromatography (DCM / MeOH = 50 / 1) was performed with a rotary evaporator to obtain 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).

[0176] (4) Synthesis of intermediate 13-4

[0177] Intermediate 13-3 (0.1 g, 0.24 mmol) was dissolved in 2 mL of DMAC solution, and NCS (35 mg, 0.26 mmol) was added. The reaction was carried out at 100 °C for 0.5 h. The reaction was monitored by TLC to be almost complete. The mixture was extracted with EA and water, the organic phase was washed with saturated sodium chloride and dried with anhydrous sodium sulfate, and then separated by column chromatography (DCM / MeOH = 20:1) to obtain 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).

[0178] (5) Synthesis of compound 13

[0179] Referring to step (2) of Example 11, intermediate 11-2 was replaced with intermediate 13-4 to obtain compound 13 (8 mg, 18.3%). 1 H NMR (400MHz, DMSO-d6) 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).

[0180] Example 14 Synthesis of Compound 14

[0181] [Reaction Route 12]

[0182]

[0183] (1) Synthesis of intermediate 14-1

[0184] Intermediate 13-2 (0.2 g, 0.49 mmol) was dissolved in 1 ml of DMSO solution, and 2 ml of methanol and K2CO3 (0.06 g, 0.49 mmol) were added. The mixture was microwaved at 80 °C for 1 h. The reaction was monitored by TLC and found to be almost complete. The mixture was extracted with EA and water, the organic phase was washed with saturated sodium chloride and dried with anhydrous sodium sulfate, and then separated by column chromatography (DCM / MeOH = 20:1) to obtain 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).

[0185] (2) Synthesis of intermediate 14-2

[0186] Referring to step (4) of Example 13, intermediate 13-3 was replaced with 14-1 to obtain intermediate 14-2 (0.06g, 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).

[0187] (3) Synthesis of compound 14

[0188] Referring to step (2) of Example 11, intermediate 11-2 was replaced with intermediate 14-2 to obtain 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=8.8H z,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).

[0189] Example 15 Synthesis of Compound 15

[0190] Referring to step (1) of Example 11, intermediate 11-1 was replaced with 4-amino-2-fluorobenzonitrile (purchased from Bidex) to obtain intermediate 15-1. 1 H NMR (400MHz, DMSO-d6) 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).

[0191] Referring to step (2) of Example 11, intermediate 11-2 was replaced with intermediate 15-1 to obtain compound 15. 1 H 1 HNMR(400MHz,DMSO-d6)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).

[0192] Example 16 Synthesis of Compound 16

[0193] Following the method of Example 12, intermediate 11-2 was replaced with intermediate 15-1 to obtain compound 16.

[0194] 1 H NMR (400MHz, DMSO-d6) δ11.28(s,1H),10.45(s,1H),7.89(s,1H),7.72(s,1H),7.58(d,J=8.6Hz,1H),7.02–6.9 7(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).

[0195] Example 17 Synthesis of Compound 17

[0196] [Reaction Route 13]

[0197]

[0198] (1) Synthesis of intermediate 17-2

[0199] Intermediate 17-1 (0.5 g, 2.7 mmol) was dissolved in 20 ml of methanol, and palladium on carbon (0.1 g) with a water content of 60% was added. After purging with hydrogen three times, the reaction was continued at room temperature for 7 h. The reaction was confirmed to be complete by TLC. The reaction solution was filtered and the filtrate was evaporated to dryness to obtain 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).

[0200] (2) Synthesis of intermediate 17-3

[0201] Referring to step (1) of Example 11, intermediate 11-1 was replaced with intermediate 17-2 to obtain intermediate 17-3 (0.11g, 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).

[0202] (3) Synthesis of compound 17

[0203] Referring to step (2) of Example 11, intermediate 11-2 was replaced with intermediate 17-3 to obtain compound 17 (9 mg, 19.8%). 1 H NMR (600MHz, CDCl3) 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,2 H),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).

[0204] Example 18 Synthesis of Compound 18

[0205] Referring to Example 12, intermediate 11-2 was replaced with intermediate 17-3 to obtain compound 18 (25 mg, 53%). 1HNMR (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).

[0206] Example 19 Synthesis of Compound 19

[0207] Referring to step (1) of Example 11, intermediate 11-1 was replaced with intermediate 6-amino-2-chlorocyanopyridine to obtain 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).

[0208] Referring to step (2) of Example 11, intermediate 11-2 was replaced with intermediate 19-1 to obtain compound 19. 1 H NMR (400MHz, CDCl3) 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,2 H),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).

[0209] Example 20 Synthesis of Compound 20

[0210] Referring to Example 12, intermediate 11-2 was replaced with intermediate 19-1 to obtain compound 20. 1H 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).

[0211] Example 21 Synthesis of Compound 21

[0212] [Reaction Route 14]

[0213]

[0214] (1) Synthesis of intermediate 21-1

[0215] 5-Bromo-2-hydrazinopyridine (2 g, 10.6 mmol) was dissolved in tetrahydrofuran, and N,N'-carbonyldiimidazole (2.6 g, 15.9 mmol) was added at room temperature. The reaction mixture was heated to reflux overnight. The reaction was found to be nearly complete by TLC. The mixture was extracted with ethyl acetate and water, the organic layer was washed with saturated sodium chloride, and dried with anhydrous sodium sulfate. The crude intermediate 21-1 (1.3 g, 58%) was obtained by rotary evaporation.

[0216] (2) Synthesis of intermediate 21-2

[0217] Intermediate 21-1 (1.3 g, 6.07 mmol) was dissolved in 15 mL of phosphorus oxychloride and heated to 110 °C overnight. The reaction was found to be nearly complete by TLC. The reaction solution was cooled and slowly added dropwise to ice water. Extraction was performed with ethyl acetate and water. The organic layer was washed with saturated sodium bicarbonate and saturated sodium chloride, and dried with anhydrous sodium sulfate. The solution was then evaporated to dryness and subjected to column chromatography (DCM / MeOH = 20 / 1) to obtain intermediate 21-2 (0.86 g, 61%). 1 H NMR (400MHz, CDCl3) δ8.17 (s, 1H), 7.67 (d, J = 9.7Hz, 1H), 7.38 (dd, J = 9.7, 1.6Hz, 1H).

[0218] (3) Synthesis of intermediate 21-3

[0219] Compound 21-2 (0.86 g, 3.69 mmol) was dissolved in benzylamine (5 ml) and reacted overnight at 110 °C. The reaction solution was extracted with ethyl acetate and water, the organic layer was washed with saturated sodium chloride and dried with anhydrous sodium sulfate, and then subjected to column chromatography (DCM / MeOH = 20 / 1) to give intermediate 21-3.

[0220] (4) Synthesis of intermediate 21-4

[0221] Intermediate 21-3 was dissolved in DMSO, and potassium carbonate, L-proline and cuprous iodide were added. The reaction solution was stirred at room temperature for 5 minutes under nitrogen protection, then ammonia was added and the reaction solution was heated to 90°C overnight. TLC showed that the reaction was basically complete. The solution was extracted with ethyl acetate and water, the organic layer was washed with saturated sodium chloride and dried with anhydrous sodium sulfate, evaporated to dryness and separated by column chromatography to obtain intermediate 21-4.

[0222] (5) Synthesis of intermediate 21-5

[0223] Compound 21-4 was dissolved in 50 ml of methanol, palladium on carbon with a water content of 60% was added, hydrogen was purged three times, and the reaction was continued at room temperature for 7 h. The reaction was confirmed to be complete by TLC. The reaction solution was filtered, and the filtrate was evaporated to dryness to obtain intermediate 21-5.

[0224] (6) Synthesis of compound 21

[0225] Referring to step (9) of Example 1, intermediates 1-6 were replaced with intermediates 21-5 to obtain compound 21.

[0226] Example 22 Synthesis of Compound 22

[0227] Referring to step (9) of Example 1, intermediates 1-6 were replaced with 4-aminophthalimide to obtain 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).

[0228] Example 23 Synthesis of Compound 23

[0229] [Reaction Route 15]

[0230]

[0231] (1) Synthesis of intermediate 23-1

[0232] 4-Chloro-7-nitroquinazoline (0.5 g, 2.39 mmol) was dissolved in 20 mL of ammonia in methanol (7N) and reacted at room temperature for 2 h. The reaction was confirmed to be complete by TLC. The reaction solution was then evaporated to dryness to obtain 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).

[0233] (2) Synthesis of intermediate 23-2

[0234] Intermediate 23-1 (0.3 g, 0.9 mmol) was dissolved in 30 ml of methanol, and palladium on carbon (0.06 g) with a water content of 60% was added. After purging with hydrogen three times, the reaction was continued at room temperature for 7 h. The reaction was confirmed to be complete by TLC. The reaction solution was filtered, and the filtrate was evaporated to dryness 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).

[0235] (3) Synthesis of compound 23

[0236] Referring to step (9) of Example 1, intermediates 1-6 were replaced with intermediates 23-2 to obtain compound 23 (12 mg, 11%). 1 H NMR (400MHz, CDCl3) 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).

[0237] Example 24 Synthesis of Compound 24

[0238] Following the synthetic method of Example 23, 4-chloro-7-nitroquinazoline was replaced with 4-chloro-6-nitroquinazoline to obtain compound 24 (61 mg, 56%). 1H NMR (400MHz, CDCl3) 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).

[0239] Example 25 Synthesis of Compound 25

[0240] [Reaction Route 16]

[0241]

[0242] (1) Synthesis of intermediate 25-1

[0243] 4-Chloro-7-nitroquinazoline (0.5 g, 2.39 mmol) was dissolved in methanol (20 ml), and sodium hydride (60% distributed in kerosene) (0.14 g, 3.6 mmol) was added under ice bath conditions. The reaction was carried out overnight at room temperature. TLC showed that the reaction was almost complete. The reaction was quenched with ice water, extracted with EA and water, and the organic layer was washed with saturated sodium bicarbonate and sodium chloride, respectively. The mixture was dried over anhydrous sodium sulfate and subjected to rotary 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).

[0244] (2) Synthesis of intermediate 25-2

[0245] Intermediate 25-1 (0.4 g, 1.94 mmol) was dissolved in 50 mL of methanol, and palladium on carbon (0.08 g) with a water content of 60% was added. After purging with hydrogen three times, the reaction was continued at room temperature for 7 h. The reaction was confirmed to be complete by TLC. The reaction solution was filtered and the filtrate was evaporated to dryness to obtain 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).

[0246] (3) Synthesis of intermediate 25-3

[0247] Referring to step (9) of Example 1, intermediate 1-6 was replaced with intermediate 25-2 to obtain intermediate 25-3 (0.09g, 61%).

[0248] (4) Synthesis of compound 25

[0249] Intermediate 25-3 (0.09 g, 0.2 mmol) was dissolved in DMF (5 ml), and lithium chloride (0.04 g, 1.0 mmol) and p-toluenesulfonic acid (0.17 g, 1.0 mmol) were added. The reaction solution was heated to 80 °C and reacted for about 3 h. TLC showed that the reaction was basically complete. After cooling, the mixture was extracted with EA and water. The organic layer was washed with saturated sodium bicarbonate and sodium chloride, respectively, dried over anhydrous sodium sulfate, and purified by rotary cyclohexane 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).

[0250] Example 26 Synthesis of Compound 26

[0251] [Reaction Route 17]

[0252]

[0253] (1) Synthesis of intermediate 26-1

[0254] 3-Hydroxy-1,2-benzisoxazole (purchased from Bidex) (1.0 g, 7.4 mmol) was dissolved in 10 ml of concentrated sulfuric acid, transferred to an ice bath, and potassium nitrate (0.74 g, 7.4 mmol) was added. The reaction was carried out for 3 h under ice bath conditions. TLC showed that the reaction was basically complete. The reaction solution was slowly added to ice water, and a large amount of white solid precipitated out. The solid was filtered to obtain a filter cake, which was dried to obtain intermediate 26-1 (1.3 g, 97.5%). 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).

[0255] (2) Synthesis of intermediate 26-2

[0256] Intermediate 26-1 (0.4 g, 2.2 mmol) was dissolved in 30 mL of dichloromethane. The reaction solution was placed in an ice-salt bath and 2-(trimethylsilyl)ethoxymethyl chloride (0.74 g, 4.4 mmol) was added under nitrogen protection. After stirring for 5 min, triethylamine (0.67 g, 6.7 mmol) was slowly added dropwise. After reacting in an ice-salt bath for 2 h, the reaction was detected by TLC to be complete. The mixture was extracted with DCM and water. The organic layer was washed with saturated sodium chloride and dried with anhydrous sodium sulfate. The residue was then separated by column chromatography (PE / EA = 10 / 1) to obtain intermediate 26-2 (0.28 g, 41%). 1 H NMR (400MHz, CDCl3) 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).

[0257] (3) Synthesis of intermediate 26-3

[0258] Intermediate 26-2 (0.28 g, 0.9 mmol) was dissolved in 100 ml of methanol, and palladium on carbon (0.05 g) with a water content of 60% was added. After purging with hydrogen three times, the reaction was continued at room temperature for 1 h. The reaction was confirmed to be complete by TLC. The reaction solution was filtered, the filtrate was evaporated to dryness, and the intermediate 26-3 (0.07 g, 28%) was obtained by column chromatography (DCM / MeOH = 20 / 1). 1 H NMR (400MHz, CDCl3) 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).

[0259] (4) Synthesis of intermediate 26-4

[0260] Referring to step (9) of Example 1, intermediate 1-6 is replaced with intermediate 26-3 to obtain intermediate 26-4. 1H NMR(400MHz, CDCl3)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).

[0261] (5) Synthesis of compound 26

[0262] Intermediate 26-4 (50 mg, 0.09 mmol) was dissolved in dichloromethane, and 1 ml of trifluoroacetic acid was added under ice bath conditions. After stirring at room temperature for 1 h, the reaction was detected by TLC to indicate that the reaction was complete. The reaction solution was evaporated to dryness and separated by column chromatography (DCM / MeOH = 20 / 1) to obtain 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).

[0263] Example 27 Synthesis of Compound 27

[0264] [Reaction Route 18]

[0265]

[0266] (1) Synthesis of intermediate 27-1

[0267] 3,6-Dichloropyridazine-4-carboxylic acid (0.5 g, 2.59 mmol), compounds 1-3 (0.534 g, 3.11 mmol), and potassium carbonate (1.79 g, 12.95 mmol) were dissolved in 30 mL of DMF and reacted overnight at 80 °C. After the reaction was confirmed to be complete by TLC, 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 with anhydrous sodium sulfate, and evaporated to dryness to give crude product 27-1 (0.3 g, 39.7%). 1H NMR (600MHz, DMSO-d6) 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).

[0268] (2) Synthesis of compound 27

[0269] Intermediate 27-1 (0.10 g, 0.34 mmol) was dissolved in anhydrous DCM (5 ml). The reaction solution was placed in an ice bath, and 2-3 drops of DMF were added. Oxaloyl chloride (2 M, 0.2 ml) was added under nitrogen protection. After reacting in an ice bath for 2 h, the reaction solution was evaporated to dryness to obtain intermediate 27-2.

[0270] Add 1-6 (0.061 g, 0.41 mmol) to the reaction flask of intermediate 27-2 and place it in an ice bath. Add 5 mL of anhydrous pyridine. After 2 h, the reaction was detected by TLC to be complete. Extract with EA and water. Wash the organic layer with saturated sodium chloride and dry with anhydrous sodium sulfate. Rotate dry and perform column chromatography (DCM / MeOH = 20 / 1) to give product 27 (0.03 g, 17.3%). 1 H NMR(400MHz,DMSO-d6)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).

[0271] Test Example Bioactivity Test

[0272] 1. Detection Method: Whole-cell manual patch-clamp technique was used to detect the effect of compounds on the voltage-gated Nav1.8 channel current. 2. Preparation and Analysis of Detected Compounds

[0273] Negative control: Electrophysiological extracellular fluid containing 0.5% DMSO

[0274] Positive control: VX-150 is the positive control drug.

[0275] Test Compound: Weigh a certain mass of the compound and dissolve it in DMSO to prepare a 20 mM DMSO stock solution. On the day of testing, serially dilute the 20 mM stock solution with extracellular fluid to the final concentration required for detection, ensuring that the DMSO content in the test drug solution does not exceed 0.5%. This concentration of DMSO has no effect on the detected Nav1.8 channel current. For example, to prepare 100 nM and 1 μM compound solutions, the serial dilution method is as follows: First, add 5 μL of the DMSO stock solution to 10 mL of extracellular fluid and dissolve thoroughly to obtain a 10 μM compound solution; then, add 1 mL of the 10 μM compound to 9 mL of extracellular fluid and dissolve thoroughly to obtain a 1 μM compound solution; finally, add 1 mL of the 1 μM compound to 9 mL of extracellular fluid and dissolve thoroughly to obtain a 100 nM compound solution.

[0276] 3. Cell Culture

[0277] (1) Nav1.8 cell line: HEK293 (Flp-In T-Rex-293) cells that stably express human Nav1.8 sodium channel, with the following encoding gene information: NM_001293306.2.

[0278] (2) Culture and passage conditions and methods: Cell lines were cultured in a constant temperature incubator at 37℃ and 5% CO2. Nav1.8 stable transgenic cells were cultured in complete medium containing 10% tetracycline-free fetal bovine serum (HyClone) and 100 μg / mL Hygromycin B in DMEM (Gibco) high glucose. The day before the experiment, when the cells reached about 90% confluence, they were digested and passaged. First, the medium was aspirated, and the cells were washed with phosphate-buffered saline (PBS) preheated to 37℃. After discarding the PBS buffer, trypsin was added for digestion, and the cells were transferred to centrifuge tubes. The cells were centrifuged at 800 rpm for 3 minutes, the supernatant was discarded, and the cells were resuspended in complete medium containing 1 μg / mL Doxcycline. The cells were then passaged into 6-well plates and induced for 20 hours. After that, the cells were isolated and passaged into coverslips coated with poly-L-lysine and cultured for another 1-2 hours before being used for electrophysiological recording experiments.

[0279] 4. Electrophysiological experiments

[0280] (1) Nav1.8 sodium channel currents were recorded at room temperature (23-25°C) using whole-cell voltage clamp technique.

[0281] (2) Whole-cell voltage-clamp recording experiments were performed using an Axon patch 700B patch-clamp amplifier (Molecular Devices), a Digidata 1440A digital-to-analog converter (Molecular Devices), and glass microelectrodes were drawn from glass electrode blanks (World Precision Instruments) using a drawing machine (P97, Sutter). The tip resistance after perfusion with electrode fluid was approximately 1.5-2.5 MΩ. The glass microelectrodes were connected to the patch-clamp amplifier by inserting them into the amplifier probe. Clamp voltage and data recording were controlled and recorded by computer using pClamp 10 software (Molecular Devices), with a sampling frequency of 20 kHz and a filtering frequency of 2 kHz.

[0282] (3) Extracellular and intracellular fluids used in electrophysiological experiments:

[0283] Extracellular fluid formulation: 140mM NaCl, 3mM KCl, 1mM CaCl2, 1mM MgCl2, 10mM HEPES and 20mM glucose, adjusted to pH 7.3 with NaOH.

[0284] Intracellular fluid formulation: 140mM CsF, 10mM NaCl, 10mM HEPES, 1.1mM EGTA and 20mM glucose, pH adjusted to 7.3 with CsOH.

[0285] Abbreviations: HEPES: 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid, N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid); EGTA: ethylene glycol bis(2-aminoethyl ether)tetraacetic acid; all drugs were purchased from Sigma.

[0286] (4) Electrophysiological stimulation protocol: After obtaining whole-cell recordings, hold the cells at -80mV for 4-5 minutes until the intracellular fluid and electrode fluid reach equilibrium, then begin electrophysiological recording (achieving high-impedance GΩ sealing conditions). Current stimulation and compound activity assay protocol: Hold the cells at -80mV, apply a 20ms depolarization voltage of +10mV, then repolarize to -80mV at a stimulation frequency of 0.5Hz. Once the Nav1.8 sodium channel current has stabilized (approximately 1 minute), begin the drug administration process until the cell current no longer changes (compound inhibition reaches steady state). Test at least 3 cells (n≥3) for each compound concentration. After all compounds are tested, administer a single concentration of 100nM VX-150 as a positive control.

[0287] 5. Data Analysis

[0288] Data acquisition, analysis, and processing were performed using pClamp10 (Molecular Devices), GraphPad Prism 5 (GraphPad Software), and Excel (Microsoft). All data are expressed as mean ± standard error (Mean ± SEM). The effect of the compound on the current was calculated using the following formula:

[0289] Inhibition rate (%) = [1 - magnitude of current after drug administration (I)] Drug ) / Magnitude of current before drug administration (I) Control )]×100.

[0290] The dose-response curve was fitted using the Hill equation: Y = Bottom + (Top - Bottom) / (1 + 10^(LogIC)) 50 -X)×k), where Bottom and Top represent the minimum and maximum values ​​of inhibition, respectively, X represents the logarithm of the compound concentration, and Y represents I Drug / I Control Value, IC 50 This represents the drug dose that produces the half-maximal inhibitory effect, and k represents the Hill coefficient.

[0291] The results are shown in Tables 1 and 2.

[0292] Table 1. Activity IC of the compounds in the examples on the Nav1.8 channel 50 value

[0293]

[0294]

[0295] Table 2 shows the percentage blocking activity of the compounds in the examples on the Nav1.8 channel.

[0296]

[0297]

[0298] The above embodiments are merely illustrative of the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein, without departing from the spirit and substance defined by the claims of the present invention; and such modifications or substitutions are still within the scope defined by the claims of the present invention.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, in, Compounds of Formula I are selected from the following compounds:

2. A method for preparing the compound according to claim 1, comprising the following steps: Compound of formula I was obtained by acylation of formulas III and IV in the presence of a base.

3. The preparation method according to claim 2, wherein, The base is selected from pyridine, sodium carbonate, and sodium bicarbonate.

4. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable excipient.

5. Use of a compound of claim 1 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of claim 4 in the preparation of a Nav1.8 inhibitor.

6. Use of a compound of claim 1 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of claim 4 in the preparation of a medicament for treating, preventing or controlling diseases or symptoms associated with the Nav1.8 channel.

7. The use according to claim 6, wherein, The diseases or symptoms associated with the Nav1.8 channel include nociceptive pain, inflammatory pain, neuropathic pain, functional pain, muscle or skeletal injury-related pain, pelvic pain, abdominal pain, chest pain, lumbosacral neuralgia, preoperative pain, intraoperative pain, postoperative pain, migraine, trigeminal neuralgia, pancreatitis, renal colic, cancer pain, pain caused by chemotherapy or drug therapy, diabetic neuropathy, postherpetic neuralgia, back pain, phantom limb pain, sciatica, small fiber neuralgia, erythromelalgia, arthritis, pruritus, asthma, multiple sclerosis, arrhythmia, atrial fibrillation, heart failure, Brugada syndrome, kidney stones, epilepsy, and seizures.

8. The use according to claim 6, wherein, The diseases or symptoms associated with the Nav1.8 channel are acute or chronic pain, and acute or chronic itching.

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