Cis-enaminoamide-pyrazole derivatives and their use in anti-inflammatory drugs

Abstract

The application discloses cis-olefin amide-pyrazole derivatives and application thereof in anti-inflammatory drugs. The compounds have the effects of anti-inflammation, treatment of lung injury, chronic obstructive pulmonary disease and the like.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to cis-enamides-pyrazole derivatives, which have the effects of anti-inflammation, treatment of pneumonia and lung injury. BACKGROUND

[0002] Pneumonia is an important symptom of many diseases in the lung, such as chronic obstructive pulmonary disease and acute respiratory distress syndrome, which is mainly manifested as diffuse inflammation aggregation in the lung, and the pathological manifestations are damage to alveolar epithelial cells and capillary endothelial cells, and diffuse pulmonary interstitial edema and alveolar edema. The causes of pneumonia mainly include infection of bacteria, viruses, rickettsia, mycoplasma and fungi. When pollutants or bacteria enter the lung through the respiratory tract, they will deposit in the alveolar cells, be phagocytosed by macrophages and release cytokines to attract immune cells and neutrophils, thereby stimulating the lung epithelial cells to release inflammatory factors such as IL-6, TNF-alpha and IL-1 beta to cause inflammation, and the sustained inflammatory response further damages the lung cells, leading to reduced lung function and inducing pro-inflammatory cascade, thereby causing lung injury.

[0003] The lead compound N-methyl-N-cis-styryl-cinnamic amide (lansiumamide B) is a cis-enamide compound isolated from the fruit core of Lansium parasiticum, which has various pharmacological activities such as treatment of obesity, anti-diabetes, antibacterial and anti-inflammatory. However, the cis-trans configuration of the enamide in the structure of lansiumamide B affects its biological activity, and the biological activity of the cis structure is obviously stronger than that of the trans structure. Pyrazole, the English name, is a nitrogen-containing aromatic compound, which is often used as a building block of drug molecules in medicinal chemistry. Pyrazole and its derivatives have various pharmacological activities such as antibacterial, antiviral, antitumor, anti-inflammatory and antidepressant, and are therefore widely used in the medical field. For example, pyrazole compounds have potential application value in the treatment of cancer, cardiovascular disease, dementia and diabetes. The present application aims to combine the cis-enamide structure with the common pyrazole structure in medicinal chemistry to develop anti-inflammatory drugs for treating pneumonia and lung injury. SUMMARY

[0004] The purpose of the present application is to provide cis-enamides-pyrazole derivatives and their use in the preparation of anti-inflammatory drugs.

[0005] One or more embodiments of the present application provide a compound of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, co-crystal or deuteride thereof.

[0006]

[0007] R1is C6-14 aryl, five- to fourteen-membered heteroaryl, C6-10 cycloalkyl, five- to fourteen-membered heterocyclyl, C1-C6 alkyl, C1-C6 cycloalkyl, halogen, cyano; optionally, the C6-14 aryl, five- to fourteen-membered heteroaryl, C6-10 cycloalkyl, or five- to fourteen-membered heterocyclyl is substituted with one or more substituents selected from the group consisting of halogen, halogenated C1-C6 alkyl, carbonyl, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, carboxyl, cyano, hydroxyl, carbonyl, phenyl-SO2-, nitro, C1-C6 alkyl oxycarbonyl; the five- to fourteen-membered heteroaryl or five- to fourteen-membered heterocyclyl comprises 1-3 heteroatoms selected from the group consisting of N, O and S; preferably, the halogen is F, Cl, Br or I;

[0008] R2is H, C1-C6 alkyl, C1-C6 alkoxy, halogen, C1-C6 halogenated alkyl, C1-C6 halogenated alkoxy;

[0009] In one or more embodiments, when the amide group is at the 5-position of the pyrazole, R1is at the 3-position of the pyrazole.

[0010] In one or more embodiments, when the amide group is at the 3-position of the pyrazole, R1is at the 5-position of the pyrazole.

[0011] In one or more embodiments, Formula I is wherein each R1is independently t-butyl, methyl, phenyl, and R2is hydrogen, fluorine, chlorine, bromine, methyl, methoxy, trifluoromethyl, trifluoromethoxy.

[0012] In one or more embodiments, Formula I is wherein each R1is independently methyl, t-butyl, phenyl, and R2is hydrogen, fluorine, chlorine, bromine, methyl, methoxy, trifluoromethyl, trifluoromethoxy.

[0013] In one or more embodiments, Formula I is each R1is independently

[0014] One or more embodiments of the present application provide a compound or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, co-crystal, or deuteride thereof:

[0015]

[0016] One or more embodiments of the present application provide a method of preparing a compound of Formula I,

[0017]

[0018] wherein R1, R2 are as described above;

[0019] The preparation method comprises: (1)

[0021]

[0022] a) adding the compounds of formula I and formula II to a sodium ethoxide solution at -20°C to 40°C, and reacting at 20°C to 50°C for 8 to 12 hours;

[0023] b) adding the product compound of formula III obtained in step a) to a compound of formula IV in anhydrous methanol at 0°C, and stirring at room temperature for 1 to 2 hours, and then reacting at 60°C to 90°C for 2 to 4 hours;

[0024] c) dissolving the product compound of formula V obtained in step b) in an ammonium methanol solution, and reacting at 20°C to 50°C for 2 to 8 days;

[0025] d) reacting the product compound of formula VI obtained in step c) with a compound of formula VII at 40°C to 80°C for 0.5 to 3 hours;

[0026] e) stirring the product compound of formula VIII obtained in step d) with sodium hydride at -20°C to 5°C for 0.5 to 1 hour, then adding methyl iodide at the temperature, and finally stirring at 20°C to 50°C for 2 to 6 hours to obtain a compound of formula IX; (2)

[0028]

[0029] f) adding the product compound of formula III obtained in step a) to a compound of formula IV in anhydrous ethanol at 0°C, and stirring at room temperature for 1 to 2 hours, and then reacting at 60°C to 90°C for 5 to 10 hours;

[0030] g) dissolving the product compound of formula X obtained in step f) in an ammonium methanol solution, and reacting at 20°C to 50°C for 3 to 9 days;

[0031] h) reacting the product compound of formula XI obtained in step g) with a compound of formula VII at 40°C to 80°C for 1 to 4 hours;

[0032] i) stirring the product compound of formula XII obtained in step h) with sodium hydride at -20°C to 10°C for 0.5 to 1 hour, then adding methyl iodide at the temperature, and finally stirring at 20°C to 50°C for 3 to 7 hours to obtain a compound of formula XIII; (3)

[0034]

[0035] j) stirring the compound of formula IV, the compound of formula XIV and methyl propiolate at 0 °C for 5 minutes, then adding potassium carbonate, potassium bromide and potassium peroxomonosulfate (Oxone), and stirring the reaction at 20-40 °C for 9-12 hours;

[0036] k) dissolving the product of step j), the compound of formula XV, in a solution of ammonium hydroxide in methanol and reacting at 20-50 °C for 3-10 days;

[0037] l) reacting the product of step k), the compound of formula XVI, with the compound of formula VII at 40-80 °C for 1-4 hours;

[0038] m) stirring the product of step 1), the compound of formula XVII, with sodium hydride at -20-10 °C for 0.5-1 hour, then adding methyl iodide at this temperature, and finally stirring the reaction at 20-50 °C for 3-7 hours to obtain the compound of formula XVIII;

[0039] wherein R1, R2 are as described in any one of claims 1-5.

[0040] One or more embodiments of the present application provide a pharmaceutical composition comprising a compound of the present application and a pharmaceutically acceptable excipient or diluent.

[0041] One or more embodiments of the present application provide use of a compound of the present application or a pharmaceutical composition of the present application in the manufacture of a medicament for preventing and / or treating an inflammatory disease, a respiratory disease.

[0042] One or more embodiments of the present application provide a compound of the present application for use as a medicament.

[0043] One or more embodiments of the present application provide a pharmaceutical composition of the present application for use as a medicament.

[0044] One or more embodiments of the present application provide a compound or composition of the present application for use in preventing and / or treating an inflammatory disease, a respiratory disease.

[0045] One or more embodiments of the present application provide a compound or composition of the present application for use in preventing and / or treating an IL-1β, IL-6, TNF-α mediated disease.

[0046] One or more embodiments of the present application provide a compound or composition of the present application for use in inhibiting IL-1β, IL-6, TNF-α.

[0047] One or more embodiments of the present application provide methods of preventing and / or treating inflammatory diseases, respiratory diseases, the methods comprising administering to a subject in need thereof a compound or a composition of the present application.

[0048] One or more embodiments of the present application provide methods of preventing and / or treating IL-1β, IL-6, TNF-α mediated diseases, the methods comprising administering to a subject in need thereof a compound or a composition of the present application.

[0049] One or more embodiments of the present application provide methods of inhibiting IL-1β, IL-6, TNF-α, the methods comprising administering to a subject in need thereof a compound or a composition of the present application.

[0050] In one or more embodiments, the inflammatory disease is an inflammatory lung disease.

[0051] In one or more embodiments, the respiratory disease is acute lung injury or pneumonia.

[0052] One or more embodiments of the present application provide uses of a compound of the present application or a pharmaceutical composition of the present application in the manufacture of a medicament for preventing and / or treating IL-1β, IL-6, TNF-α mediated diseases or an IL-1β, IL-6, TNF-α inhibitor.

[0053] In one or more embodiments, the compound of the present application has anti-inflammatory and therapeutic uses for psoriasis. Biological experiments demonstrate that the compound of the present application can significantly reduce the number of inflammatory cells in an inflammation model, or reduce the level of IL-1β, IL-6, TNF-α in cells.

[0054] In one or more embodiments, the results of lung injury model mouse experiments show that the compound of the present application can significantly improve the severity of mouse alveolar atrophy, inflammatory infiltration and fibrous collagen formation, indicating that the compound of the present application can reduce the release of pro-inflammatory factors, reduce the production of pro-inflammatory mediators, to inhibit the occurrence and development of inflammatory response, and also can protect the mouse lung, to alleviate the effect of lung injury.

[0055] The following explains the terms used in the technical solutions of the present application. As used in the specification and the appended claims, unless otherwise indicated, the terms of the present application have the following meanings:

[0056] The term "halogen" means fluorine, chlorine, bromine or iodine.

[0057] The term "amino" means -NH2.

[0058] The term "hydroxyl" means -OH.

[0059] "Alkyl" refers to a linear or branched saturated aliphatic hydrocarbon group of 1 to 20 carbon atoms, preferably an alkyl group of 1 to 8 (e.g., 1, 2, 3, 4, 5, 6, 7, 8) carbon atoms, more preferably an alkyl group of 1 to 6 carbon atoms, and even more preferably an alkyl group of 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, neobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, and various branched isomers thereof; when an alkyl group is substituted, it may optionally be further substituted with one or more substituents.

[0060] "Alkoxy" refers to a group in which at least one carbon atom in an alkyl group is replaced by an oxygen atom. Non-limiting examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, n-hexyloxy, cyclopropyloxy, and cyclobutyloxy. The definition of "alkyl" is the same as that of "alkyl" described above.

[0061] "Aryl" refers to a substituted or unsubstituted aromatic ring, which can be, for example, a monocyclic ring of 6 to 8 carbon atoms (e.g., 6, 7, 8 carbon atoms), a bicyclic ring of 6 to 12 carbon atoms (e.g., 6, 7, 8, 9, 10, 11, 12 carbon atoms), or a tricyclic ring system of 10 to 14 carbon atoms (e.g., 10, 11, 12, 13, 14 carbon atoms), which can be a bridged ring or a spirocyclic ring, non-limiting examples of which include phenyl and naphthyl. The aryl group can optionally be further substituted with one or more substituents.

[0062] "Heteroaryl" refers to a substituted or unsubstituted aromatic ring, which can be, for example, a 5- to 8-membered (e.g., 5, 6, 7, 8-membered) monocyclic ring, a 5- to 12-membered (e.g., 5, 6, 7, 8, 9, 10, 11, 12-membered) bicyclic ring, or a 10- to 14-membered (e.g., 10, 11, 12, 13, 14-membered) tricyclic ring system, and contains 1 to 6 (e.g., 1, 2, 3, 4, 5, 6) heteroatoms selected from N, O or S, preferably a 5- to 8-membered heteroaryl group, and the 1 to 4 (e.g., 1, 2, 3, 4) N and S optionally substituted in the heteroaryl ring can be oxidized to various oxidation states. The heteroaryl group may be attached to a heteroatom or a carbon atom, and may be a bridged ring or a spirocyclic ring. Non-limiting examples include cyclic pyridyl, furyl, thienyl, pyranyl, pyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, piperidinylbenzimidazolyl, benzopyridinyl, and pyrrolopyridinyl. The heteroaryl group may be further substituted with one or more substituents.

[0063] "Heterocyclyl" or "heterocycle" means a saturated or unsaturated non-aromatic heterocycle, which for example can be a 5- to 10-membered (e.g., 5-, 6-, 7-, 8-, 9-, 10-membered) monocyclic, 5- to 12-membered (e.g., 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-membered) bicyclic, or 10- to 14-membered (e.g., 10-, 11-, 12-, 13-, 14-membered) tricyclic ring system, and contains 1 to 4 (e.g., 1, 2, 3, 4) heteroatoms selected from N, O, or S, preferably a 5- to 8-membered heterocyclyl. Optionally 1 to 4 (e.g., 1, 2, 3, 4) of the N, S in the ring of the "heterocyclyl" or "heterocycle" can be oxidized into various oxidation states; the "heterocyclyl" or "heterocycle" can be attached at a heteroatom or carbon atom; the "heterocyclyl" or "heterocycle" can be a bridged or spirocyclic ring. Non-limiting examples of "heterocyclyl" or "heterocycle" include oxiranyl, oxetanyl, aziridinyl, oxetanyl, thietanyl, 1,3-dioxolanyl, 1,4-dioxolanyl, 1,3-dioxananyl, azepanyl, oxepanyl, thiepanyl, dioxepanyl, pyridinyl, piperidinyl, homopiperidinyl, furanyl, thienyl, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, piperazinyl, homopiperazinyl, imidazolyl, pyrrolidinyl, morpholinyl, thiomorpholinyl, thioxanyl, 1,3-dithianyl, dihydrofuranyl, dihydropyranyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydropyrrolyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydropyranyl, benzimidazolyl, benzopyridinyl, pyrrolopyridinyl, benzodihydrofuranyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydrothienyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 1,2,3,4-tetrahydroisoquinolinyl, 3-azabicyclo[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl, azabicyclo[2.2.2]hexanyl, 3H-indolizinyl, quinolizinyl, N-pyridinylurea, 1,1-dioxothiomorpholinyl, azabicyclo[3.2.1]octanyl, azabicyclo[5.2.0]nonyl, oxatricyclo[5.3.1.1]dodecanyl, azadamantanyl, and oxaspiro[3.3]heptanyl. The "heterocyclyl" or "heterocycle" can be optionally further substituted with one or more substituents.

[0064] "Cycloalkyl" means a saturated cyclic hydrocarbon group, which for example can be a 6- to 10-carbon atom (e.g., 6-, 7-, 8-, 9-, 10-carbon atom) monocyclic, bicyclic, or polycyclic ring, preferably 6- to 8-carbon atom ring. Non-limiting examples of "cycloalkyl" include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. When the cycloalkyl is substituted, it can be optionally further substituted with one or more substituents.

[0065] "Pharmaceutically acceptable salt" or "pharmaceutically acceptable salt thereof means a salt of a compound of the present application which retains the biological effectiveness and properties of the free acids or free bases and is obtained by reaction of the free acid with a non-toxic inorganic or organic base, or the free base with a non-toxic inorganic or organic acid.

[0066] "Pharmaceutical composition" means a mixture of one or more compounds of the present application, pharmaceutically acceptable salts or prodrugs thereof, and other chemical components, such as pharmaceutically acceptable carriers, excipients, and / or one or more other therapeutic agents.

[0067] "Carrier" means a material that does not itself induce the production of antibodies to it, and which does not have an adverse effect on the activity and properties of a given compound.

[0068] "Excipient" means an inert substance added to a pharmaceutical composition to facilitate administration of a compound. Non-limiting examples include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives, gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, binders, and disintegrating agents.

[0069] "Prodrug" means a compound that is converted into a biologically, pharmaceutically or therapeutically active compound of the present application after administration to a subject. Prodrugs of the present application are prepared by modifying the amino or carboxyl groups of the compounds of the present application in such a way that the modifications are cleaved in vivo to give the parent compound. When a prodrug of the present application is administered to a mammalian subject, the prodrug is cleaved to form the free amino or carboxyl groups.

[0070] "Cocrystal" means a crystal formed by the combination of an active pharmaceutical ingredient (API) and a cocrystal former (CCF) under the action of hydrogen bonds or other non-covalent bonds, wherein both the pure state of the API and the pure state of the CCF are solids at room temperature, and there is a fixed stoichiometric ratio between the components. Cocrystals are a type of multi-component crystal, including binary cocrystals formed between two neutral solids, and multi-component cocrystals formed between a neutral solid and a salt or a solvate.

[0071] "Stereoisomer" means isomers that differ in the orientation of atoms in space, including enantiomers and diastereomers.

[0072] "Optional" or "optionally" or "selective" or "selectively" means that the subsequently described event or circumstance can or can not occur, and this description includes instances in which the event or circumstance occurs and instances in which it does not. For example, "heterocyclyl optionally substituted with alkyl" means that the alkyl group can or can not be present, and this description includes instances in which the heterocyclyl is substituted with alkyl, and instances in which the heterocyclyl is not substituted with alkyl. BRIEF DESCRIPTION OF DRAWINGS

[0073] Figure 1 Representative inhibition of the cis-enaminoamide-pyrazole derivatives on Raw264.7 cell toxicity and on NO level, IL-6, IL-1 β inflammatory factor mRNA level in active example 1, compared with the control group * P<0.05, ** P<0.01, *** P<0.001; and compared with the model group # P<0.05, ## P<0.01, ### P<0.001.

[0074] Figure 2 Representative inhibition of the compound on inflammatory factor IL-6, IL-1 β protein level in Raw264.7 cell model in active example 2, compared with the control group **** P<0.0001; and compared with the LPS group ## P<0.01, #### P<0.0001.

[0075] Figure 3 Representative change of the body weight of mice in active example 3.

[0076] Figure 4 Representative total number of inflammatory cells in the alveolar lavage fluid of mice and total number of white blood cells in the whole blood in active example 3, compared with the control group **** P<0.0001; and compared with the model group # P<0.05, ## P<0.01, #### P<0.0001.

[0077] Figure 5 Representative HE staining pathological graph of the lung tissue of mice in active example 3, magnification: 200x; Scale bar = 200 μm.

[0078] Figure 6 Representative Masson staining graph of the lung tissue of mice in active example 3, magnification: 200x; Scale bar = 200 μm.

[0079] Figure 7 Representative immunohistochemical staining of macrophage iNOS, Arg-1 and neutrophil LY6G marker graph of the lung tissue of mice in active example 3. DETAILED DESCRIPTION

[0080] The application will be further described in conjunction with specific examples. These examples are for the purpose of illustration only and do not limit the scope and spirit of the application. Based on the examples in the application, all other examples obtained by those of ordinary skill in the art without creative work are within the scope of protection of the application.

[0081] Example 1 Synthesis of a series of compounds

[0082] (1) Synthesis of compound 2

[0083]

[0084] Take 100 mL of a sealed tube and add anhydrous ethanol (6 mL), slowly drop 20% sodium ethoxide solution (2.1 mL, 10 mmol), ice water bath cooling. Pinacolone (1.0 g, 10 mmol), diethyl oxalate (1.46 g, 10 mmol) are mixed in advance, and then added dropwise to the pre-cooled sodium ethoxide solution. After the dropwise addition is completed, continue to react for 12 h after the reaction solution is restored to room temperature. TLC monitoring tracking until the raw material disappears or no change, under ice water bath with 20% dilute sulfuric acid to pH = 3, diatomite filtration, and washed with dichloromethane (3 x 20 mL), the aqueous phase is extracted with dichloromethane (3 x 20 mL). After the combined organic phase is washed with saturated sodium chloride solution (20 mL), dry over anhydrous sodium sulfate for 30 minutes, concentrated under reduced pressure, and the obtained crude product is separated and purified by silica gel chromatography column, using petroleum ether and ethyl acetate (V / V = 5:1) as eluent to obtain yellow liquid product 1.4 g (compound 2), with a yield of 70%.

[0085] (2) Synthesis of compound 3

[0086]

[0087] Anhydrous methanol (5 mL) is added to a 15 mL sealed tube, and then phenylhydrazine (0.324 g, 3 mmol) is added. After being cooled in an ice bath, methyl 5,5-dimethyl-2,4-dioxohexanoate (0.60 g, 3.0 mmol) is slowly added, and the reaction is stirred at room temperature for 1 h and then refluxed for 2 h. After TLC monitoring until the raw material disappears or no change, the oil bath is removed, and the reaction solution is naturally cooled to room temperature. The organic phase is collected with ethyl acetate, concentrated under reduced pressure, and the obtained crude product is separated and purified by silica gel chromatography column, using petroleum ether and ethyl acetate (V / V = 10:1) as eluent to obtain yellow solid product 0.416 g, with a yield of 51%.

[0088] (3) Synthesis of compound 4

[0089]

[0090] Into a 15 mL sealed tube was added ethyl 5-tert-butyl-l-phenylpyrazole-3-carboxylate (0.544 g, 2 mmol), 7 mol / L ammonia methanol solution (4 mL), and the reaction was stirred at room temperature for 7 d. After TLC monitoring until the starting material disappeared or no change, the organic phase was collected with acetone, and concentrated under reduced pressure. The obtained crude product was separated and purified on a silica gel chromatographic column with petroleum ether and acetone (V / V = 1:1) as eluent to obtain a white solid product 0.46 g with a yield of 94%.

[0091] (4) Synthesis of compound 5

[0092]

[0093] Into a dry 10 mL Schlenk tube was filled with argon, compound 4 (243 mg, 1 mmol), cuprous iodide (9.5 mg, 0.05 mmol), cesium carbonate (407 mg, 1.25 mmol), and (Z)-2-iodovinylbenzene (253 mg, 1.1 mmol) was added to the reaction tube after being diluted with dry anhydrous tetrahydrofuran (5 mL), and then N,N'-dimethylethylenediamine (DMEDA, 10.8 μL, 0.1 mmol) was added dropwise. The reaction was continued at 65°C in an oil bath for 3 h under sealed conditions. After natural cooling, it was filtered with diatomite, washed with ethyl acetate (3 x 20 mL), and the organic phase was dried with anhydrous sodium sulfate for 30 min before being filtered off. The filtrate was concentrated under reduced pressure, and the obtained crude product was separated and purified on a silica gel column to obtain a light yellow solid 259 mg with a yield of 75%.

[0094] (5) Synthesis of compound A1

[0095]

[0096] Into a dry two-necked flask was filled with argon, compound 5 (173 mg, 0.5 mmol) was dissolved in dry anhydrous N,N-dimethylformamide (2 mL), and an ice water bath was used for cooling. Then 60% sodium hydride (32 mg, 0.8 mmol) was added, and the reaction was continued at the same temperature for 30 min before adding iodomethane (142 mg, 1 mmol) and reacting for 3 h. After the reaction was completed, ice water was added, and the reaction mixture was extracted with ethyl acetate (3 x 20 mL) and washed with saturated sodium chloride solution (20 mL). The organic phase was dried with anhydrous sodium sulfate for 30 min before being filtered off. The filtrate was concentrated under reduced pressure, and the obtained crude product was separated and purified on a silica gel column to obtain a yellow oil 160 mg (compound A1) with a yield of 89%.

[0097] The obtained compound A1 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were as follows:

[0098] Major conformational isomers: 1H NMR (400 MHz, CDC13) δ: 7.44-7.37 (m, 3H), 7.33-7.30 (m, 2H), 7.26-7.20 (m, 4H), 7.17-7.13 (m, 1H), 6.87 (d, J = 8.8 Hz, 1H), 6.43 (s, 1H), 5.86 (d, J = 8.8 Hz, 1H), 3.02 (s, 3H), 1.09 (s, 9H). 13 C NMR (100 MHz, CDC13) δ: 164.6, 153.7, 145.7, 141.9, 135.6, 130.2, 129.5 (2 x C), 128.8 (2 x C), 128.7 (2 x C), 128.4 (2 x C), 127.3 (2 x C), 118.9, 107.3, 38.8, 32.1, 30.7 (3 x C). HRMS (ESI) m / z calcd for C 1 H NMR (400 MHz, CDC13) δ: 7.44-7.37 (m, 3H), 7.33-7.30 (m, 2H), 7.26-7.20 (m, 4H), 7.17-7.13 (m, 1H), 6.93 (br, 1H), 6.67 (s, 1H), 6.05 (br, 1H), 3.22 (s, 3H), 1.09 (s, 9H). 13 C NMR (100 MHz, CDC13) δ: 164.6, 153.7, 145.7, 141.9, 135.6, 130.2, 129.5 (2 x C), 128.8 (2 x C), 128.7 (2 x C), 128.4 (2 x C), 127.3 (2 x C), 118.9, 107.3, 38.8, 32.1, 30.7 (3 x C). HRMS (ESI) m / z calcd for C 23 H 26 ON3 + [M+H] + 360.2070, found 360.2074.

[0099] By selecting different methyl ketones and aryl hydrazines, compounds A1-A10 can be obtained. The compounds A1-A10 are arranged in order as follows:

[0100]

[0101] Synthesis of compound A2 in Example 2:

[0102] The synthesis step is referred to Example 1, and the hydrazine reagent of the second step reaction is selected as p-fluorophenylhydrazine to obtain compound A2, yield: 88%; major conformational isomer: 1H NMR (400 MHz, CDC13) δ: 7.38-7.35 (m, 2H), 7.33-7.26 (m, 4H), 7.24-7.21 (m, 1H), 7.15 (t, J = 8.4 Hz, 2H), 6.90 (d, J = 8.8 Hz, 1H), 6.50 (s, 1H), 5.94 (d, J = 8.8 Hz, 1H), 3.10 (s, 3H), 1.16 (s, 9H). 13 C NMR (100 MHz, CDC13) δ: 164.4, 162.9 (d, J = 248 Hz), 153.9, 145.8, 137.8 (d, J = 3.4 Hz), 135.5, 131.2, 130.5 (d, J = 8.8 Hz, 2 x C), 128.7 (2 x C), 128.4 (2 x C), 127.3, 119.5, 115.7 (d, J = 23 Hz, 2 x C), 106.5, 35.7, 32.0, 30.6 (3 x C). 19 F NMR (376 MHz, CDC13) δ: -111.1. Minor conformer: 1 H NMR (400 MHz, CDC13) δ: 7.38-7.35 (m, 2H), 7.33-7.26 (m, 4H), 7.24-7.21 (m, 1H), 7.15 (t, J = 8.4 Hz, 2H), 6.90 (d, J = 8.8 Hz, 1H), 6.50 (s, 1H), 5.94 (d, J = 8.8 Hz, 1H), 3.10 (s, 3H), 1.16 (s, 9H). 13 C NMR (100 MHz, CDC13) δ: 164.4, 162.9 (d, J = 248 Hz), 153.9, 145.8, 137.8 (d, J = 3.4 Hz), 135.5, 131.2, 130.5 (d, J = 8.8 Hz, 2 x C), 128.7 (2 x C), 128.4 (2 x C), 127.3, 118.9, 115.7 (d, J = 23 Hz, 2 x C), 107.4, 38.8, 32.0, 26.9 (3 x C). 19 F NMR (376 MHz, CDC13) δ: -111.1. HRMS (ESI) m / z calcd for C 23 H 25 ON3F + [M+H] + 378.1976, found 378.1981.

[0103] Synthesis of compound A3 of Example 3:

[0104] The synthesis procedure was referred to Example 1, and the hydrazine reagent of the second step reaction was selected as p-chlorobenzylhydrazine to obtain compound A3 in a yield of 86%; major conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.46-7.44 (m, 2H), 7.34-7.22 (m, 7H), 6.90 (d, J = 8.8 Hz, 1H), 6.51 (s, 1H), 5.94 (d, J = 8.8 Hz, 1H), 3.10 (s, 3H), 1.17 (s, 9H). 13 C NMR (100 MHz, CDC13) δ: 164.3, 153.9, 146.0, 140.4, 135.5, 131.2, 130.1 (3 x C), 129.0 (2 x C), 128.7 (2 x C), 128.4 (2 x C), 127.3, 119.6, 106.7, 35.8, 32.0, 30.7 (3 x C). Minor conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.46-7.44 (m, 2H), 7.34-7.22 (m, 7H), 6.90 (d, J = 8.8 Hz, 1H), 6.51 (s, 1H), 5.94 (d, J = 8.8 Hz, 1H), 3.10 (s, 3H), 1.17 (s, 9H). 13 C NMR (100 MHz, CDC13) δ: 164.3, 153.9, 146.0, 140.4, 135.5, 131.2, 130.1 (3 x C), 129.0 (2 x C), 128.7 (2 x C), 128.4 (2 x C), 127.3, 119.6, 106.7, 35.8, 32.0, 30.7 (3 x C). Minor conformational isomer: 23 H 25 ON3Cl + [M+H] + 394.1681, found 394.1684.

[0105] Synthesis of compound A4 in Example 4:

[0106] The synthesis procedure was referred to Example 1, and the hydrazine reagent of the second step reaction was selected as p-chlorobenzylhydrazine to obtain compound A3 in a yield of 86%; major conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.46-7.44 (m, 2H), 7.34-7.22 (m, 7H), 6.90 (d, J = 8.8 Hz, 1H), 6.51 (s, 1H), 5.94 (d, J = 8.8 Hz, 1H), 3.10 (s, 3H), 1.17 (s, 9H). 13C NMR (100 MHz, CDC13) δ: 164.4, 153.9, 146.1, 141.0, 135.6, 132.0 (2 x C), 131.2, 130.4 (2 x C), 128.8 (2 x C), 128.5 (2 x C), 127.4, 123.6, 119.1, 107.6, 38.9, 32.1, 30.7 (3 x C). HRMS (ESI) m / z calcd for C 1 H NMR (400 MHz, CDC13) δ: 7.61 (d, J = 8.4 Hz, 2H), 7.34 - 7.22 (m, 7H), 7.02 (br, 1H), 6.75 (s, 1H), 6.14 (br, 1H), 3.28 (s, 3H), 1.18 (s, 9H). 13 C NMR (100 MHz, CDC13) δ: 164.4, 153.9, 146.1, 141.0, 135.6, 132.0 (2 x C), 131.2, 130.4 (2 x C), 128.8 (2 x C), 128.5 (2 x C), 127.4, 123.6, 119.1, 107.6, 38.9, 32.1, 30.7 (3 x C). HRMS (ESI) m / z calcd for C 23 H 25 ON3Br + [M+H] + 438.1176, found 438.1187.

[0107] Synthesis of compound A5

[0108] Synthesis step refer to example 1, the hydrazine reagent of the second step reaction is selected to be p-tolylhydrazine, to obtain compound A5, yield: 84%; major conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.34 - 7.22 (m, 9H), 6.97 (d, J = 8.8 Hz, 1H), 6.51 (s, 1H), 5.93 (d, J = 8.8 Hz, 1H), 3.10 (s, 3H), 2.45 (s, 3H), 1.17 (s, 9H). 13 C NMR (100 MHz, CDC13) δ: 164.4, 153.9, 146.1, 141.0, 135.6, 132.0 (2 x C), 131.2, 130.4 (2 x C), 128.8 (2 x C), 128.5 (2 x C), 127.4, 123.6, 119.1, 107.6, 38.9, 32.1, 30.7 (3 x C). HRMS (ESI) m / z calcd for C 1H NMR (400 MHz, CDC13) δ: 7.34-7.22 (m, 9H), 6.97 (d, J = 8.8 Hz, 1H), 6.74 (s, 1H), 6.14 (br, 1H), 3.30 (s, 3H), 2.45 (s, 3H), 1.17 (s, 9H). 13 C NMR (100 MHz, CDC13) δ: 164.7, 153.7, 145.6, 139.6, 139.4, 135.7, 130.2, 129.3 (2 x C), 128.8 (2 x C), 128.5 (2 x C), 128.4 (2 x C), 127.2, 120.4, 107.3, 39.0, 32.1, 30.7 (3 x C), 21.4. HRMS (ESI) m / z calcd for C 24 H 28 ON3 + [M+H] + 374.2227, found 374.2229.

[0109] Synthesis of compound A6:

[0110] Synthesis step refer to example 1, the hydrazine reagent of the second step reaction is selected to be p-methoxyphenylhydrazine, to obtain compound A6, yield: 96%; major conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.34-7.21 (m, 7H), 6.96 (d, J = 8.8 Hz, 3H), 6.50 (s, 1H), 5.93 (d, J = 8.8 Hz, 1H), 3.88 (s, 3H), 3.10 (s, 3H), 1.17 (s, 9H). 13 C NMR (100 MHz, CDC13) δ: 164.7, 160.2, 153.9, 145.6, 135.7, 134.7 (2 x C), 131.4, 129.9 (2 x C), 128.8 (2 x C), 128.4 (2 x C), 127.2, 119.1, 113.8, 106.4, 55.7, 35.8, 32.1, 30.7 (3 x C). Minor conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.34-7.21 (m, 7H), 6.96 (d, J = 8.8 Hz, 3H), 6.73 (s, 1H), 6.13 (br, 1H), 3.88 (s, 3H), 3.30 (s, 3H), 1.17 (s, 9H). 13C NMR (100 MHz, CDC13) δ: 164.7, 160.2, 153.9, 145.6, 135.7, 134.7 (2 x C), 130.2, 129.9 (2 x C), 128.8 (2 x C), 128.4 (2 x C), 127.2, 118.9, 113.8, 107.2, 55.7, 38.8, 32.1, 30.7 (3 x C). HRMS (ESI) m / z calcd for C 24 H 28 O2N3 + [M+H] + 390.2176, found 390.2177.

[0111] Synthesis of compound A7

[0112] Synthesis procedure was referred to Example 1, the hydrazine reagent of the second step reaction was chosen as p-trifluoromethylphenylhydrazine, to obtain compound A7, yield: 85%; major conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.76 (d, J = 8.0 Hz, 2H), 7.54 (d, J = 8.0 Hz, 2H), 7.35-7.22 (m, 5H), 6.89 (d, J = 8.8 Hz, 1H), 6.55 (s, 1H), 5.96 (d, J = 8.8 Hz, 1H), 3.11 (s, 3H), 1.19 (s, 9H). 13 C NMR (100 MHz, CDC13) δ: 164.3, 154.1, 146.4, 145.0, 135.5, 131.7 (q, J = 32.7 Hz), 131.1, 129.3 (2 x C), 128.8 (2 x C), 128.5 (2 x C), 127.4, 126.1, 126.0, 123.7 (q, J = 270 Hz), 119.8, 107.0, 35.8, 32.1, 30.8 (3 x C). 19 F NMR (376 MHz, CDC13) δ: -62.6 (3 x F). Minor conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.76 (d, J = 8.0 Hz, 2H), 7.54 (d, J = 8.0 Hz, 2H), 7.35-7.22 (m, 5H), 7.02 (br, 1H), 6.79 (s, 1H), 6.16 (br, 1H), 3.29 (s, 3H), 1.19 (s, 9H). 13C NMR (100 MHz, CDC13) δ: 164.3, 154.3, 146.6, 145.0, 135.5, 131.7 (q, J = 32.7 Hz), 130.1, 129.3 (2 x C), 128.8 (2 x C), 128.5 (2 x C), 127.4, 126.1, 126.0, 123.7 (q, J = 270 Hz), 119.2, 107.8, 38.9, 32.1, 30.8 (3 x C). 19 F NMR (376 MHz, CDC13) δ: -62.6 (3 x F). HRMS (ESI) m / z calcd for C 24 H 25 ON3F3 + [M+H] + 428.1944, found 428.1947.

[0113] Synthesis of compound A8 of Example 8:

[0114] Synthesis step was referred to Example 1, the hydrazine reagent of the second step reaction was selected to be p-trifluoromethoxyphenylhydrazine, to obtain compound A8, yield: 87%; major conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.41 (d, J = 8.8 Hz, 2H), 7.31-7.20 (m, 7H), 6.87 (d, J = 8.8 Hz, 1H), 6.49 (s, 1H), 5.93 (d, J = 8.8 Hz, 1H), 3.09 (s, 3H), 1.15 (s, 9H). 13 C NMR (100 MHz, CDC13) δ: 164.4, 154.0, 149.7, 146.2, 140.3, 135.5, 131.2, 130.4 (2 x C), 128.8 (2 x C), 128.5 (2 x C), 127.4, 121.1 (2 x C), 120.4 (q, J = 257 Hz), 119.7, 106.7, 35.8, 32.1, 30.7 (3 x C). 19 F NMR (376 MHz, CDC13) δ: -57.9 (3 x F). Minor conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.41 (d, J = 8.8 Hz, 2H), 7.31-7.20 (m, 7H), 6.87 (d, J = 8.8 Hz, 1H), 6.49 (s, 1H), 5.93 (d, J = 8.8 Hz, 1H), 3.09 (s, 3H), 1.15 (s, 9H). 13C NMR (100 MHz, CDC13) δ: 164.4, 154.0, 149.7, 146.2, 140.3, 135.8, 130.1, 130.4 (2 x C), 128.8 (2 x C), 128.5 (2 x C), 127.4, 121.1 (2 x C), 120.4 (q, J = 257 Hz), 119.7, 107.6, 38.8, 32.1, 30.7 (3 x C). 19 F NMR (376 MHz, CDC13) δ: -57.9 (3 x F). HRMS (ESI) m / z calcd for C 24 H 25 02N3F3 + [M + H] + 444.1893, found 444.1903.

[0115] Synthesis of compound A9 of Example 9:

[0116] Synthesis steps refer to Example 1, the methyl ketone reagent of the first step reaction is selected as acetone, and the hydrazine reagent of the second step reaction is selected as phenylhydrazine to obtain compound A9, yield: 87%; major conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.49-7.44 (m, 4H), 7.42-7.38 (m, 1H), 7.34-7.29 (m, 4H), 7.24-7.20 (m, 1H), 6.96 (d, J = 8.8 Hz, 1H), 6.62 (s, 1H), 5.98 (d, J = 8.8 Hz, 1H), 3.09 (s, 3H), 2.34 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 164.4, 146.9, 139.5, 135.5, 131.5, 129.2 (2 x C), 128.8 (2 x C), 128.4 (2 x C), 128.2, 127.4, 125.0 (3 x C), 119.2, 109.5, 35.8, 12.5. Minor conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.49-7.44 (m, 4H), 7.42-7.38 (m, 1H), 7.34-7.29 (m, 4H), 7.24-7.20 (m, 1H), 7.03 (br, 1H), 6.73 (s, 1H), 6.13 (br, 1H), 3.34 (s, 3H), 2.34 (s, 3H). 13C NMR (100 MHz, CDC13) δ: 164.4, 146.9, 139.5, 135.8, 130.1, 129.2 (2 x C), 128.8 (2 x C), 128.4 (2 x C), 128.2, 127.4, 125.0 (3 x C), 119.0, 110.1, 39.0, 12.5. HRMS (ESI) m / z calcd for C 20 H 20 ON3 + [M+H] + 318.1601, found 318.1611.

[0117] Synthesis of compound A10

[0118] The synthesis was performed according to the procedure described in Example 1, with the exception that the methyl ketone reagent for the first reaction was chosen to be acetophenone and the hydrazine reagent for the second reaction was chosen to be phenylhydrazine to give compound A10 in 95% yield. Major conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.37-7.32 (m, 12H), 7.28-7.25 (m, 3H), 7.04 (d, J = 8.8 Hz, 1H), 6.92 (s, 1H), 6.06 (d, J = 8.8 Hz, 1H), 3.16 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 164.1, 147.4, 143.6, 139.7, 135.5, 131.4, 129.9, 129.0 (2 x C), 128.9 (2 x C), 128.8 (2 x C), 128.7 (2 x C), 128.6 (2 x C), 128.5, 128.0, 127.5, 125.3 (2 x C), 119.8, 110.2, 35.9. Minor conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.37-7.32 (m, 12H), 7.28-7.25 (m, 3H), 7.08 (s, 1H), 7.04 (d, J = 8.8 Hz, 1H), 6.19 (br, 1H), 3.45 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 164.4, 147.4, 143.6, 139.7, 135.8, 131.4, 130.1, 129.0 (2 x C), 128.9 (2 x C), 128.8 (2 x C), 128.7 (2 x C), 128.6 (2 x C), 128.5, 128.0, 127.5, 125.3 (2 x C), 119.2, 111.0, 39.1. HRMS (ESI) m / z calcd for C 25H 22 ON3 + [M+H] + 380.1757, found 380.1768.

[0119] Example 11 Synthesis of compounds of series B

[0120] (1) Synthesis of compound 6

[0121]

[0122] Take 100 mL of sealed tube and add anhydrous ethanol (6 mL), slowly drop 20% sodium ethoxide solution (2.1 mL, 10 mmol), ice water bath cooling. Acetone (0.58 g, 10 mmol), diethyl oxalate (1.46 g, 10 mmol) were mixed in advance, then added to the pre-cooled sodium ethoxide solution. After the dropwise addition was completed, the reaction was continued for 12 h after the temperature was restored to room temperature. TLC monitoring tracking until the starting material disappeared or no change, under ice water bath with 20% dilute sulfuric acid acidification to pH = 3, diatomite filtration, and washed with dichloromethane (3 x 20 mL), the aqueous phase was extracted with dichloromethane (3 x 20 mL). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate for 30 minutes, concentrated under reduced pressure, and the obtained crude product was separated and purified by silica gel column chromatography using petroleum ether and ethyl acetate (V / V = 5:1) as eluent to obtain yellow liquid product 1.14 g (compound 6) with a yield of 72%.

[0123] (2) Synthesis of compound 7

[0124]

[0125] Anhydrous ethanol (5 mL), phenylhydrazine (324 mg, 3 mmol) were added to a 15 mL sealed tube, and compound 6 (475 mg, 3 mmol) was slowly added. The reaction was maintained at 80°C for 5 h. After TLC monitoring until the starting material disappeared or no change, the oil bath was removed, and the reaction solution was naturally cooled to room temperature. The organic phase was collected with ethyl acetate, and concentrated under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography using petroleum ether and ethyl acetate (V / V = 10:1) as eluent to obtain yellowish solid product 317 mg with a yield of 46%.

[0126] (3) Synthesis of compound 8

[0127]

[0128] Into a 15 mL vial, compound 7 (460 mg, 2 mmol), 7 mol / L ammonia methanol solution (4 mL) were added, and the reaction was stirred at room temperature for 7 d. After TLC monitoring until the starting material disappeared or no change, the organic phase was collected with acetone, and concentrated under reduced pressure. The obtained crude product was separated and purified on a silica gel column, and separated with petroleum ether and acetone (V / V = 1:1) as eluent to obtain white solid product 370 mg, with a yield of 92%.

[0129] (4) Synthesis of compound 9

[0130]

[0131] Into a dry 10 mL Schlenk tube, argon was filled, compound 8 (201 mg, 1 mmol), cuprous iodide (9.5 mg, 0.05 mmol), cesium carbonate (407 mg, 1.25 mmol) were added, and then (Z)-2-iodovinylbenzene (253 mg, 1.1 mmol) was added after being diluted with dry anhydrous tetrahydrofuran (5 mL), and then DMEDA (10.8 μL, 0.1 mmol) was added dropwise. The reaction was continued at 65°C oil bath for 3 h under the condition of sealing. After natural cooling, it was filtered with diatomite, washed with ethyl acetate (3×20 mL), and the organic phase was dried with anhydrous sodium sulfate for 30 min, then filtered out, and the filtrate was concentrated under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain 170 mg of light yellow solid with a yield of 56%.

[0132] (5) Synthesis of compound B1

[0133]

[0134] Into a dry two-necked flask, argon was filled, compound 9 (152 mg, 0.5 mmol) dissolved in dry anhydrous N,N-dimethylformamide (2 mL) was added, and then an ice water bath was cooled. Sodium hydride (32 mg, 0.8 mmol) was added, and then the reaction was continued at the same temperature for 30 min. Then, iodomethane (142 mg, 1 mmol) was added and the reaction was continued for 3 h. After the reaction was completed, ice water was added, and then extracted with ethyl acetate (3×20 mL). The organic phase was washed with saturated sodium chloride solution (20 mL), and then dried with anhydrous sodium sulfate for 30 min, then filtered out, and the filtrate was concentrated under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain 137 mg of yellow oil (compound B1) with a yield of 86%.

[0135] The obtained compound B1 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were as follows:

[0136] Major conformational isomers: 1H NMR (400 MHz, CDC13) δ: 7.54-7.37 (m, 4H), 7.33-7.30 (m, 1H), 7.25-7.09 (m, 3H), 6.86-6.79 (m, 2H), 6.31 (s, 1H), 6.01 (d, J = 9.2 Hz, 1H), 5.82 (d, J = 9.2 Hz, 1H), 2.93 (s, 3H), 2.32 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 163.1, 149.5, 140.0, 137.3, 134.6, 129.3 (2 x C), 128.6 (2 x C), 128.4 (3 x C), 127.9, 127.8, 123.8 (2 x C), 121.2, 109.0, 34.9, 13.6. Minor conformer: 1 H NMR (400 MHz, CDC13) δ: 7.54-7.37 (m, 4H), 7.33-7.30 (m, 1H), 7.25-7.09 (m, 3H), 6.95 (d, J = 9.6 Hz, 1H), 6.86-6.79 (m, 2H), 6.42 (s, 1H), 6.09 (d, J = 9.6 Hz, 1H), 2.51 (s, 3H), 2.37 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 164.3, 150.2, 139.9, 137.1, 135.1, 129.8 (2 x C), 128.9 (2 x C), 128.3 (3 x C), 127.9, 127.4, 123.2 (2 x C), 119.7, 108.0, 37.7, 13.6. HRMS (ESI) m / z calcd for C 20 H 20 ON3 + [M+H] + 318.1601, found 318.1604.

[0137] By selecting different methyl ketones and aryl hydrazines, compounds B1-B10 can be obtained.

[0138] The compounds B1-B10 are arranged in the following order:

[0139]

[0140] Synthesis of compound B2 in Example 12:

[0141] The synthesis step is referred to Example 11, and the hydrazine reagent of the second step reaction is selected as p-fluorophenylhydrazine to obtain compound B2 with a yield of 80%; 1H NMR (400 MHz, CDCI3) δ: 7.41-7.38 (m, 2H), 7.31-7.25 (m, 3H), 7.12-7.07 (m, 2H), 6.95-6.93 (m, 2H), 6.36 (s, 1H), 6.11 (d, J = 8.8 Hz, 1H), 5.90 (d, J = 8.8 Hz, 1H), 2.98 (s, 3H), 2.33 (s, 3H). 13 C NMR (100 MHz, CDCI3) δ: 162.8, 162.0 (d, J = 246 Hz), 149.4, 137.3, 136.2 (d, J = 2.7 Hz), 134.4, 128.6 (2 x C), 128.5 (2 x C), 128.4, 128.0, 125.9 (d, J = 8.6 Hz, 2 x C), 121.7, 116.1 (d, J = 22.8 Hz, 2 x C), 109.1, 35.0, 13.5. 19 F NMR (376 MHz, CDCI3) δ: -113.8. HRMS (ESI) m / z calcd for C 20 H 19 ON3F + [M+H] + 336.1507, found 336.1513.

[0142] Synthesis of compound B3 of example 13:

[0143] Synthesis step refer to example 11, the hydrazine reagent of the second step reaction is selected to be p-chlorophenylhydrazine, to obtain compound B3, yield: 93%; major conformational isomer: 1 H NMR (400 MHz, CDCI3) δ: 7.37 (s, 4H), 7.31-7.20 (m, 3H), 6.92-6.90 (m, 2H), 6.35 (s, 1H), 6.07 (d, J = 8.8 Hz, 1H), 5.89 (d, J = 8.8 Hz, 1H), 2.99 (s, 3H), 2.33 (s, 3H). 13 C NMR (100 MHz, CDCI3) δ: 162.7, 149.7, 138.5, 137.2, 134.4, 133.6, 129.4 (2 x C), 128.5 (4 x C), 128.3, 128.0, 125.1 (2 x C), 121.9, 109.3, 35.0, 13.5. minor conformational isomer: 1H NMR (400 MHz, CDC13) δ: 7.51 (s, 4H), 7.31-7.20 (m, 3H), 6.95 (d, J = 8.8 Hz, 1H), 6.92-6.90 (m, 2H), 6.44 (s, 1H), 6.15 (d, J = 8.8 Hz, 1H), 2.56 (s, 3H), 2.38 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 164.0, 150.4, 138.5, 137.2, 135.1, 133.6, 129.8 (2 x C), 128.5 (4 x C), 127.9, 127.7, 124.4 (2 x C), 120.1, 108.4, 37.7, 13.5. HRMS (ESI) m / z calcd for C 20 H 19 ON3Cl + [M+H] + 352.1211, found 352.1215.

[0144] Synthesis of compound B4 in Example 14:

[0145] Synthesis step refers to Example 11, the hydrazine reagent of the second step reaction is selected as p-bromophenylhydrazine, to obtain compound B4, yield: 93%; major conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.52 (d, J = 8.8 Hz, 2H), 7.30 (d, J = 8.8 Hz, 2H), 7.31-7.19 (m, 3H), 6.91-6.89 (m, 2H), 6.35 (s, 1H), 6.06 (d, J = 8.8 Hz, 1H), 5.88 (d, J = 8.8 Hz, 1H), 2.98 (s, 3H), 2.33 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.7, 149.8, 139.0, 137.2, 134.3, 132.3 (2 x C), 128.6 (2 x C), 128.5 (2 x C), 128.3, 128.0, 125.3 (2 x C), 121.9, 121.5, 109.4, 34.9, 13.5. minor conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.66 (d, J = 8.4 Hz, 2H), 7.45 (d, J = 8.4 Hz, 2H), 7.31-7.19 (m, 3H), 6.91-6.89 (m, 2H), 6.44 (s, 1H), 6.95 (d, J = 9.6 Hz, 1H), 6.15 (d, J = 9.6 Hz, 1H), 2.55 (s, 3H), 2.38 (s, 3H).13 C NMR (100 MHz, CDC13) δ: 163.9, 150.4, 138.3, 137.0, 135.1, 132.8 (2 x C), 128.4 (4 x C), 127.8, 127.7, 124.6 (2 x C), 121.9, 120.1, 108.5, 37.7, 13.5. HRMS (ESI) m / z calcd for C 20 H 19 ON3Br + [M+H] + 396.0706, found 396.0710.

[0146] Synthesis of compound B5 in Example 15:

[0147] Synthesis step refer to Example 11, the hydrazine reagent of the second step reaction is selected as p-tolylhydrazine, to obtain compound B5, yield: 97%; major conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.43-7.08 (m, 7H), 6.83-6.78 (m, 2H), 6.30 (s, 1H), 6.01 (d, J = 8.8 Hz, 1H), 5.80 (d, J = 8.8 Hz, 1H), 2.92 (s, 3H), 2.34 (s, 3H), 2.32 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 163.2, 149.2, 137.7, 137.6, 137.2, 134.5, 129.7 (2 x C), 128.5 (2 x C), 128.4, 128.2 (2 x C), 127.6, 123.6 (2 x C), 120.9, 108.5, 34.8, 21.1, 13.5. Minor conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.43-7.08 (m, 7H), 6.95 (d, J = 9.2 Hz, 1H), 6.83-6.78 (m, 2H), 6.40 (s, 1H), 6.08 (d, J = 9.2 Hz, 1H), 2.51 (s, 3H), 2.46 (s, 3H), 2.36 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 164.3, 149.7, 137.7, 137.4, 136.9, 135.1, 130.2 (2 x C), 128.7 (2 x C), 128.1 (2 x C), 127.9, 127.4, 123.0 (2 x C), 119.5, 107.6, 37.6, 26.9, 13.5. HRMS (ESI) m / z calcd for C 21 H22 ON3 + [M+H] + 332.1757, found 332.1764.

[0148] Synthesis of compound B6 in Example 16:

[0149] Synthesis step refers to Example 11, the hydrazine reagent of the second step reaction is selected as p-methoxyphenylhydrazine, to obtain compound B6, yield: 85%; major conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.30 (d, J = 8.8 Hz, 2H), 7.21-7.06 (m, 3H), 6.92-6.76 (m, 4H), 6.28 (s, 1H), 6.01 (d, J = 8.8 Hz, 1H), 5.79 (d, J = 8.8 Hz, 1H), 3.73 (s, 3H), 2.90 (s, 3H), 2.28 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 163.0, 159.1, 148.8, 137.2, 134.4, 133.1, 128.5 (2 x C), 128.4, 128.2 (2 x C), 127.6, 125.2 (2 x C), 120.9, 114.2 (2 x C), 108.2, 55.4, 34.7, 13.4. Minor conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.43 (d, J = 8.8 Hz, 2H), 7.21-7.06 (m, 3H), 6.99 (d, J = 8.4 Hz, 1H), 6.92-6.76 (m, 4H), 6.36 (s, 1H), 6.06 (br, 1H), 3.82 (s, 3H), 2.49 (s, 3H), 2.32 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 164.2, 159.1, 149.5, 137.0, 135.0, 132.9, 128.6 (2 x C), 128.0 (2 x C), 127.8, 127.3, 124.7 (2 x C), 119.5, 114.6 (2 x C), 107.3, 55.5, 37.5, 26.8. HRMS (ESI) m / z calcd for C 21 H 22 O2N3 + [M+H] + 348.1707, found 348.1714.

[0150] Synthesis of compound B7 in Example 17:

[0151] The synthesis step refers to Example 11, Step 2, and the hydrazine reagent is selected to be p-trifluoromethylphenylhydrazine to give compound B7 in 85% yield; major conformer: 1 H NMR (400 MHz, CDC13) δ: 7.65 (d, J = 8.4 Hz, 2H), 7.53 (d, J = 8.4 Hz, 2H), 7.28-7.20 (m, 3H), 6.96-6.88 (m, 2H), 6.38 (s, 1H), 6.07 (d, J = 8.8 Hz, 1H), 5.88 (d, J = 8.8 Hz, 1H), 2.99 (s, 3H), 2.33 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.6, 150.3, 142.7, 137.3, 134.3, 129.6 (q, J = 32.7 Hz), 128.6 (2 x C), 128.5 (2 x C), 128.2, 128.1, 126.4 (q, J = 3.6 Hz, 2 x C), 123.9 (q, J = 270 Hz), 123.7 (2 x C), 122.1, 110.1, 35.0, 13.6. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). Minor conformer: 1 H NMR (400 MHz, CDC13) δ: 7.77 (d, J = 7.6 Hz, 2H), 7.68 (d, J = 7.6 Hz, 2H), 7.18-7.12 (m, 3H), 6.96-6.88 (m, 2H), 6.95 (d, J = 8.8 Hz, 1H), 6.45 (s, 1H), 6.16 (d, J = 8.8 Hz, 1H), 2.58 (s, 3H), 2.37 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.6, 150.6, 142.2, 137.0, 134.3, 129.6 (q, J = 32.7 Hz), 128.6 (2 x C), 128.5 (2 x C), 128.2, 128.1, 126.4 (q, J = 3.6 Hz, 2 x C), 123.9 (q, J = 270 Hz), 122.9 (2 x C), 120.2, 109.1, 37.8, 27.0. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). HRMS (ESI) m / z calcd for C 21 H 19 ON3F3 + [M+H] + 386.1475, found 386.1479.

[0152] Synthesis of compound B8

[0153] Synthesis step refer to example 11, the hydrazine reagent of the second step reaction was selected to be p-trifluoromethoxyphenylhydrazine, to obtain compound B8, yield: 88%; major conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.36 (d, J = 8.8 Hz, 2H), 7.29-7.15 (m, 5H), 6.87-6.85 (m, 2H), 6.29 (s, 1H), 6.02 (d, J = 8.8 Hz, 1H), 5.81 (d, J = 8.8 Hz, 1H), 2.90 (s, 3H), 2.25 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.6, 149.8, 148.3, 138.5, 137.2, 134.4, 128.6 (2 x C), 128.5 (2 x C), 128.4, 128.0, 125.2 (2 x C), 121.8, 121.6 (2 x C), 240.9 (q, J = 256 Hz), 109.6, 35.0, 13.5. 19 F NMR (376 MHz, CDC13) δ: -57.9 (3 x F). Minor conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.51 (d, J = 7.2 Hz, 2H), 7.29-7.15 (m, 5H), 7.10 (br, 1H), 6.87-6.85 (m, 2H), 6.34 (s, 1H), 6.08 (d, J = 9.2 Hz, 1H), 2.53 (s, 3H), 2.29 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 163.9, 149.8, 148.3, 138.5, 137.2, 134.4, 128.9 (2 x C), 128.5 (2 x C), 127.8, 127.7, 124.5 (2 x C), 122.0 (2 x C), 120.1, 240.9 (q, J = 256 Hz), 108.5, 37.8, 13.5. 19 F NMR (376 MHz, CDC13) δ: -57.9 (3 x F). HRMS (ESI) m / z calcd for C 21 H 19 O2N3F3 + [M + H] + 402.1424, found 402.1428.

[0154] Synthesis of compound B9

[0155] The synthesis procedure was referred to Example 11. The methyl ketone reagent for the first step reaction was selected as 2-methyl-2-pentanone, and the hydrazine reagent for the second step reaction was selected as phenylhydrazine. Compound B8 was obtained in 98% yield; major conformer: 1 H NMR (400 MHz, CDC13) δ: 7.56-7.28 (m, 5H), 7.25-7.09 (m, 3H), 6.86-6.80 (m, 2H), 6.30 (s, 1H), 5.90 (d, J = 8.8 Hz, 1H), 5.80 (d, J = 8.8 Hz, 1H), 2.94 (s, 3H), 1.31 (s, 9H). 13 C NMR (100 MHz, CDC13) δ: 163.5, 162.6, 140.3, 136.8, 134.7, 129.3 (2 x C), 128.6 (2 x C), 128.4 (2 x C), 128.3, 127.8, 127.7, 123.8, 121.3, 105.9, 34.9, 32.4, 30.6 (3 x C). Minor conformer: 1 H NMR (400 MHz, CDC13) δ: 7.56-7.28 (m, 5H), 7.25-7.09 (m, 3H), 6.96 (d, J = 9.6 Hz, 1H), 6.86-6.80 (m, 2H), 6.49 (s, 1H), 6.09 (d, J = 9.6 Hz, 1H), 2.49 (s, 3H), 1.36 (s, 9H). 13 C NMR (100 MHz, CDC13) δ: 164.7, 163.2, 140.1, 136.5, 135.2, 129.7 (2 x C), 128.6 (2 x C), 128.4 (2 x C), 128.3, 128.0, 127.4, 123.3, 119.6, 105.0, 37.7, 32.5, 30.6 (3 x C). HRMS (ESI) m / z calcd for C 23 H 26 ON3 + [M+H] + 360.2070, found 360.2081.

[0156] Synthesis of compound B10

[0157] The synthesis procedure was referred to Example 11. The methyl ketone reagent for the first step reaction was selected as 2-methyl-2-pentanone, and the hydrazine reagent for the second step reaction was selected as phenylhydrazine. Compound B8 was obtained in 98% yield; major conformer: 1H NMR (400 MHz, CDC13) δ: 7.83 (d, J = 7.2 Hz, 2H), 7.68 - 7.33 (m, 8H), 7.24 - 7.10 (m, 3H), 6.88 - 6.81 (m, 2H), 6.76 (s, 1H), 6.01 (d, J = 8.8 Hz, 1H), 5.85 (d, J = 8.8 Hz, 1H), 2.98 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.8, 152.0, 140.1, 138.1, 134.5, 132.4, 129.4 (2 x C), 128.9, 128.8 (2 x C), 128.6 (2 x C), 128.5 (2 x C), 128.4, 128.2, 127.9, 126.0 (2 x C), 123.9 (2 x C), 121.9, 106.3, 35.0. Minor conformer: 1 H NMR (400 MHz, CDC13) δ: 7.92 - 7.88 (m, 2H), 7.68 - 7.33 (m, 8H), 7.24 - 7.10 (m, 3H), 6.98 (d, J = 9.2 Hz, 1H), 6.95 (s, 1H), 6.88 - 6.81 (m, 2H), 6.13 (d, J = 9.2 Hz, 1H), 2.56 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 164.1, 152.6, 139.9, 137.8, 135.1, 132.4, 129.8 (2 x C), 129.0, 128.9 (2 x C), 128.6 (2 x C), 128.5 (2 x C), 128.4, 128.3, 127.5, 126.0 (2 x C), 123.4 (2 x C), 119.9, 105.4, 37.7. HRMS (ESI) m / z calcd for C 25 H 22 ON3 + [M+H] + 380.1757, found 380.1767.

[0158] Synthesis of compounds of Example 21C series

[0159] (1) Synthesis of compound 10

[0160]

[0161] Into a 25 mL round bottom flask, p-fluorobenzaldehyde (205 mg, 1.65 mmol), 5 mL acetonitrile and 0.5 mL pure water, potassium carbonate (725 mg, 5.25 mmol), p-trifluoromethylphenylhydrazine hydrochloride (319 mg, 1.5 mmol), potassium bromide (214 mg, 1.8 mmol), methyl propiolate (252 mg, 3 mmol) were added in sequence, and then potassium peroxomonosulfate (1.56 g, 4.5 mmol) was added under ice-bath cooling. After 10 min, the ice-bath was removed and the reaction was stirred at room temperature. TLC monitoring was performed until the starting material was consumed. The reaction was quenched with saturated sodium sulfite solution, extracted with ethyl acetate (3 x 20 mL), washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate for 30 min, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography to obtain 350 mg (compound 10) of yellow solid with a yield of 64%. The reaction was repeated three times until sufficient amount of compound 10 was obtained.

[0162] (2) Synthesis of compound 11

[0163]

[0164] Into a 15 mL sealed tube, compound 10 (730 mg, 2 mmol), 7 mol / L ammonia methanol solution (4 mL) were added, and the reaction was stirred at room temperature for 7 d. After TLC monitoring until the starting material disappeared or showed no change, the organic phase was collected with acetone, concentrated under reduced pressure, and the obtained crude product was separated and purified on a silica gel column chromatography with petroleum ether and acetone (V / V = 1:1) as eluent to obtain 649 mg of white solid with a yield of 93%.

[0165] (3) Synthesis of compound 12

[0166]

[0167] Into a dry 10 mL Schlenk tube, argon was filled, compound 11 (349 mg, 1 mmol), cuprous iodide (9.5 mg, 0.05 mmol), cesium carbonate (407 mg, 1.25 mmol) were added, and then (Z)-2-iodovinylbenzene (253 mg, 1.1 mmol) was diluted with dry anhydrous tetrahydrofuran (5 mL) and added into the reaction tube, and then DMEDA (10.8 μL, 0.1 mmol) was added dropwise. The reaction was continued at 65 °C oil bath for 3 h under sealed condition. After natural cooling, it was filtered with diatomite, washed with ethyl acetate (3 x 20 mL), and the organic phase was dried over anhydrous sodium sulfate for 30 min, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography to obtain 325 mg of yellowish oil with a yield of 72%.

[0168] (4) Synthesis of compound C1

[0169]

[0170] A dry two necked flask was flushed with argon and compound 12 (226 mg, 0.5 mmol) dissolved in dry anhydrous N,N-dimethylformamide (2 mL) was added. The reaction mixture was cooled in an ice water bath and sodium hydride (60%) (32 mg, 0.8 mmol) was added. After 30 min at the same temperature, iodomethane (142 mg, 1 mmol) was added and the reaction was allowed to proceed for 3 h. After the completion of the reaction, ice water was added and the reaction mixture was extracted with ethyl acetate (3 x 20 mL). The organic layer was washed with saturated sodium chloride solution (20 mL) and dried over anhydrous sodium sulphate for 30 min. The filtrate was concentrated under reduced pressure and the crude obtained was purified by column chromatography on silica gel to obtain compound C1 as a colorless oil 200 mg (86% yield).

[0171] The compound C1 obtained was characterized by nuclear magnetic resonance spectroscopy and mass spectrometry techniques and the characterization results were as follows:

[0172] Major conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.84-7.80 (m, 2H), 7.72 (d, J = 8.4 Hz, 2H), 7.65 (d, J = 8.4 Hz, 2H), 7.28-7.26 (m, 3H), 7.14 (t, J = 8.8 Hz, 2H), 6.94-6.92 (m, 2H), 6.79 (s, 1H), 6.10 (d, J = 8.8 Hz, 1H), 5.95 (d, J = 8.8 Hz, 1H), 3.07 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 163.1 (d, J = 246 Hz), 162.2, 151.7, 142.6, 138.2, 134.2, 130.0 (q, J = 32.8 Hz), 128.6 (2 x C), 128.4 (2 x C), 128.2, 128.0 (d, J = 30.0 Hz, 2 x C), 127.9, 127.7 (d, J = 8.2 Hz), 126.5 (q, J = 3.6 Hz, 2 x C), 123.9 (2 x C), 123.8 (q, J = 270 Hz), 122.9, 115.8 (d, J = 21.6 Hz, 2 x C), 107.0, 35.0. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F), -112.9. Minor conformational isomer: 1 ​​​​​​​​​​​H NMR (400 MHz, CDC13) δ: 7.90-7.80 (m, 6H), 7.28-7.26 (m, 3H), 7.14 (t, J = 8.8 Hz, 2H), 6.94-6.92 (m, 2H), 7.02 (s, 1H), 7.00 (d, J = 9.2 Hz, 1H), 6.22 (d, J = 9.2 Hz, 1H), 2.66 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 163.1 (d, J = 246 Hz), 162.2, 152.3, 142.6, 137.8, 134.2, 130.0 (q, J = 32.8 Hz), 128.5 (2 x C), 128.3 (2 x C), 128.2, 128.0 (d, J = 30.0 Hz, 2 x C), 127.9, 127.7 (d, J = 8.2 Hz), 126.5 (q, J = 3.6 Hz, 2 x C), 123.9 (2 x C), 123.8 (q, J = 270 Hz), 123.1, 115.8 (d, J = 21.6 Hz, 2 x C), 106.1, 37.8. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F), -112.9. HRMS (ESI) m / z calcd for C 26 H 20 ON3F4 + [M+H] + 466.1537, found 466.1542.

[0173] By selecting different aldehydes, compounds C1-C22 can be obtained. The compounds C1-C22 are arranged in order as follows:

[0174]

[0175] Synthesis of compound C2 in Example 22:

[0176] The synthesis step is referred to Example 21, and the aldehyde of the first step reaction is selected as m-fluorobenzaldehyde to obtain compound C2, yield: 87%; major conformational isomer: 1 H NMR (400 MHz, CDC13) δ: 7.90-7.80 (m, 6H), 7.28-7.26 (m, 3H), 7.14 (t, J = 8.8 Hz, 2H), 6.94-6.92 (m, 2H), 7.02 (s, 1H), 7.00 (d, J = 9.2 Hz, 1H), 6.22 (d, J = 9.2 Hz, 1H), 2.66 (s, 3H). 13C NMR (100 MHz, CDC13) δ: 163.3 (d, J = 244 Hz), 162.1, 151.4 (d, J = 2.4 Hz), 142.6, 138.3, 134.2 (d, J = 8.1 Hz), 134.1, 130.6 (2 x C), 130.5 (q, J = 8.3 Hz), 130.0 (2 x C), 129.0, 128.3, 127.9, 126.6 (q, J = 3.6 Hz, 2 x C), 124.0 (2 x C), 123.8 (q, J = 271 Hz), 123.2, 121.7 (d, J = 2.7 Hz), 115.6 (d, J = 21.2 Hz), 112.9 (d, J = 22.7 Hz), 107.3, 35.1. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F), -112.7. Minor conformer: 1 H NMR (400 MHz, CDC13) δ: 7.84 - 7.54 (m, 6 H), 7.47 - 7.38 (m, 1 H), 7.28 - 7.06 (m, 4 H), 7.02 (s, 1 H), 6.95 - 6.92 (m, 2 H), 6.23 (d, J = 8.8 Hz, 1 H), 6.05 (d, J = 8.8 Hz, 1 H), 3.40 (s, 3 H). 13 C NMR (100 MHz, CDC13) δ: 164.6, 163.3 (d, J = 244 Hz), 151.8 (d, J = 3.2 Hz), 142.1, 138.3, 134.1, 134.0 (d, J = 8.5 Hz), 130.5 (q, J = 8.3 Hz), 129.6, 128.6 (2 x C), 128.4 (2 x C), 128.1, 127.3, 126.6 (q, J = 3.6 Hz, 2 x C), 125.2 (2 x C), 123.8 (q, J = 271 Hz), 123.4, 121.8 (d, J = 3.3 Hz), 115.8 (d, J = 23.1 Hz), 113.0 (d, J = 22.8 Hz), 107.7, 30.7. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F), -112.7. HRMS (ESI) m / z calcd for C 26 H 20 ON3F4 + [M + H] + 466.1537, found 466.1543.

[0177] Synthesis of compound C3 of Example 23:

[0178] The synthesis procedure was referred to example 21, the aldehyde of the first step reaction was selected as 2-furoylaldehyde to give compound C3 in 84% yield; 1 H NMR (400 MHz, CDC13) δ: 8.10 (td, J = 7.6, 1.6 Hz, 1H), 7.71 (d, J = 8.4 Hz, 2H), 7.64 (d, J = 8.4 Hz, 2H), 7.39-7.34 (m, 1H), 7.27-7.15 (m, 5H), 7.08 (d, J = 3.6 Hz, 1H), 6.94-6.92 (m, 2H), 6.18 (d, J = 8.8 Hz, 1H), 5.96 (d, J = 8.8 Hz, 1H), 3.06 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.2, 160.4 (d, J = 248 Hz), 147.3, 142.6, 137.7, 134.2, 130.2 (d, J = 8.4 Hz), 130.1 (q, J = 32.6 Hz), 128.7 (2 x C), 128.6, 128.5 (2 x C), 128.2, 128.1, 126.5 (q, J = 3.6 Hz, 2 x C), 124.5 (d, J = 3.2 Hz), 124.1 (2 x C), 123.9 (q, J = 271 Hz), 122.8, 119.9 (d, J = 11.5 Hz), 116.3 (d, J = 21.9 Hz), 110.6 (d, J = 10.6 Hz), 35.1. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F), -115.4. HRMS (ESI) m / z calcd for C 26 H 20 ON3F4 + [M+H] + 466.1537, found 466.1544.

[0179] Synthesis of compound C4 of example 24:

[0180] The synthesis procedure was referred to example 21, the aldehyde of the first step reaction was selected as 2-furoylaldehyde to give compound C3 in 84% yield; 1H NMR (400 MHz, CDC13) δ: 7.75 (d, J = 8.4 Hz, 2H), 7.70 (d, J = 8.4 Hz, 2H), 7.62 (d, J = 8.4 Hz, 2H), 7.39 (d, J = 8.4 Hz, 2H), 7.26-7.24 (m, 3H), 6.92-6.89 (m, 2H), 6.77 (s, 1H), 6.06 (d, J = 8.8 Hz, 1H), 5.92 (d, J = 8.8 Hz, 1H), 3.04 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.1, 151.5, 142.6, 138.3, 134.6, 134.2, 130.5, 130.1 (q, J = 32.8 Hz), 129.1 (2 x C), 128.7 (2 x C), 128.4 (2 x C), 128.2, 127.9, 127.2 (2 x C), 126.5 (q, J = 3.6 Hz, 2 x C), 123.9 (2 x C), 128.8 (q, J = 271 Hz), 123.1, 107.1, 35.1. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). HRMS (ESI) m / z calcd for C 26 H 20 ON3F3Cl + [M+H] + 482.1242, found 482.1249.

[0181] Synthesis of compound C5 of Example 25:

[0182] The synthesis step was referred to Example 21, the aldehyde of the first step reaction was selected as m-chlorobenzaldehyde, to obtain compound C5, yield: 82%; 1 H NMR (400 MHz, CDC13) δ: 7.82 (s, 1H), 7.70 (d, J = 8.4 Hz, 2H), 7.68-7.66 (m, 1H), 7.61 (d, J = 8.4 Hz, 2H), 7.37-7.32 (m, 2H), 7.26-7.24 (m, 3H), 6.93-6.90 (m, 2H), 6.78 (s, 1H), 6.06 (d, J = 8.8 Hz, 1H), 5.94 (d, J = 8.8 Hz, 1H), 3.05 (s, 3H). 13C NMR (100 MHz, CDC13) δ: 162.0, 151.2, 142.5, 138.3, 134.9, 134.2, 133.8, 130.2, 130.1 (q, J = 32.8 Hz), 128.7 (3 x C), 128.5 (2 x C), 128.3, 127.8, 126.5 (q, J = 3.6 Hz, 2 x C), 126.1, 124.1, 124.0 (2 x C), 123.8 (q, J = 271 Hz), 123.3, 107.3, 35.1. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). HRMS (ESI) m / z calcd for C 26 H 20 ON3F3Cl + [M+H] + 482.1242, found 482.1246.

[0183] Synthesis of compound C6 of example 26:

[0184] Synthesis step refer to example 21, the aldehyde of the first step reaction is selected as o-chlorobenzaldehyde, to obtain compound C6, yield: 88%; 1 H NMR (400 MHz, CDC13) δ: 7.87-7.85 (m, 1H), 7.69 (d, J = 8.4 Hz, 2H), 7.60 (d, J = 8.4 Hz, 2H), 7.48-7.45 (m, 1H), 7.36-7.29 (m, 2H), 7.26-7.23 (m, 3H), 7.16 (s, 1H), 6.96-6.94 (m, 2H), 6.20 (d, J = 8.8 Hz, 1H), 5.98 (d, J = 8.8 Hz, 1H), 3.05 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.1, 150.2, 142.6, 137.0, 134.2, 132.6, 131.0, 130.8, 130.5, 130.1 (q, J = 32.8 Hz), 129.8, 128.7 (2 x C), 128.5 (2 x C), 128.2 (2 x C), 127.1, 126.4 (q, J = 3.6 Hz, 2 x C), 124.2 (2 x C), 123.9 (q, J = 271 Hz), 123.1, 111.4, 35.1. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). HRMS (ESI) m / z calcd for C 26 H 20 ON3F3Cl + [M+H]+ 482.1242, found 482.1251.

[0185] Synthesis of compound C7:

[0186] The synthesis was performed according to the procedure described in example 21, the aldehyde of the first step reaction was chosen to be p-bromobenzaldehyde, to give compound C7 in 78% yield; 1 H NMR (400 MHz, CDC13) δ: 7.70 (d, J = 8.4 Hz, 2H), 7.69 (d, J = 8.4 Hz, 2H), 7.62 (d, J = 8.4 Hz, 2H), 7.55 (d, J = 8.4 Hz, 2H), 7.26-7.24 (m, 3H), 6.91-6.89 (m, 2H), 6.77 (s, 1H), 6.05 (d, J = 8.8 Hz, 1H), 5.92 (d, J = 8.8 Hz, 1H), 3.04 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.1, 151.5, 142.6, 138.3, 134.2, 132.0 (2 x C), 130.9, 130.1 (q, J = 32.8 Hz), 128.7 (2 x C), 128.5 (2 x C), 128.2, 127.9, 127.5 (2 x C), 126.5 (q, J = 3.6 Hz, 2 x C), 123.9 (2 x C), 123.8 (q, J = 271 Hz), 123.1, 122.8, 107.1, 35.1. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). HRMS (ESI) m / z calcd for C 26 H 20 ON3F3Br + [M+H] + 526.0736, found 526.0719.

[0187] Synthesis of compound C8:

[0188] The synthesis was performed according to the procedure described in example 21, the aldehyde of the first step reaction was chosen to be m-bromobenzaldehyde, to give compound C8 in 82% yield; 1H NMR (400 MHz, CDC13) δ: 7.98 (s, 1H), 7.81-7.69 (m, 4H), 7.61 (d, J = 8.4 Hz, 2H), 7.49 (d, J = 8.0 Hz, 1H), 7.31-7.25 (m, 3H), 6.93-6.90 (m, 2H), 6.77 (s, 1H), 6.05 (d, J = 8.8 Hz, 1H), 5.94 (d, J = 8.8 Hz, 1H), 3.05 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.0, 151.1, 142.5, 138.3, 134.2, 134.0, 131.6, 130.4, 130.1 (q, J = 32.8 Hz), 129.0, 128.7 (2 x C), 128.5 (2 x C), 128.3, 127.8, 126.6 (q, J = 3.6 Hz, 2 x C), 124.6, 124.0 (2 x C), 123.8 (q, J = 271 Hz), 123.3, 123.1, 107.3, 35.1. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). HRMS (ESI) m / z calcd for C 26 H 20 ON3F3Br + [M+H] + 526.0736, found 526.0729.

[0189] Synthesis of compound C9 of example 29:

[0190] Synthesis step refer to example 21, the aldehyde of the first step reaction is selected as o-bromobenzaldehyde, to obtain compound C9, yield: 74%; 1 H NMR (400 MHz, CDC13) δ: 7.76 (dd, J = 7.6, 1.6 Hz, 1H), 7.68 (d, J = 8.4 Hz, 2H), 7.68-7.66 (m, 1H), 7.59 (d, J = 8.4 Hz, 2H), 7.38 (t, J = 7.6 Hz, 1H), 7.26-7.21 (m, 4H), 7.14 (s, 1H), 6.98-6.96 (m, 2H), 6.21 (d, J = 8.8 Hz, 1H), 5.99 (d, J = 8.8 Hz, 1H), 3.06 (s, 3H). 13C NMR (100 MHz, CDC13) δ: 162.0, 151.6, 142.6, 136.8, 134.2, 133.8, 133.1, 131.3, 130.1 (q, J = 32.7 Hz), 130.0, 128.7 (2 x C), 128.6 (2 x C), 128.3, 128.2, 127.7, 126.4 (q, J = 3.6 Hz, 2 x C), 124.2 (2 x C), 123.9 (q, J = 271 Hz), 123.2, 122.2, 111.4, 35.1. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). HRMS (ESI) m / z calcd for C 26 H 20 ON3F3Br + [M+H] + 526.0736, found 526.0745.

[0191] Synthesis of compound C10 of Example 30:

[0192] The synthesis step was referred to Example 21, the aldehyde of the first step reaction was selected as p-tolualdehyde, to obtain compound C10, yield: 88%; 1 H NMR (400 MHz, CDC13) δ: 7.72 (d, J = 8.0 Hz, 2H), 7.69 (d, J = 8.4 Hz, 2H), 7.63 (d, J = 8.4 Hz, 2H), 7.26-7.22 (m, 5H), 6.92-6.89 (m, 2H), 6.81 (s, 1H), 6.10 (d, J = 8.8 Hz, 1H), 5.91 (d, J = 8.8 Hz, 1H), 3.03 (s, 3H), 2.39 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.4, 152.7, 142.7, 138.7, 138.0, 134.3, 129.8 (q, J = 32.8 Hz), 129.6 (2 x C), 129.1, 128.6 (2 x C), 128.5 (2 x C), 128.1, 128.0, 126.5 (q, J = 3.6 Hz, 2 x C), 125.9 (2 x C), 123.9 (2 x C), 123.8 (q, J = 271 Hz), 122.6, 107.1, 35.1, 21.5. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). HRMS (ESI) m / z calcd for C 27 H 23 ON3F3 + [M+H]+ 462.1788, found 462.1788.

[0193] Synthesis of compound C11:

[0194] The synthesis step was referred to Example 21, the aldehyde of the first step reaction was selected as m-tolualdehyde, to obtain compound C11, yield: 92%; 1 H NMR (400 MHz, CDC13) δ: 7.71-7.68 (m, 3H), 7.64-7.59 (m, 3H), 7.32 (t, J = 7.6 Hz, 1H), 7.26-7.24 (m, 3H), 7.19 (d, J = 7.6 Hz, 1H), 6.93-6.91 (m, 2H), 6.84 (s, 1H), 6.11 (d, J = 8.8 Hz, 1H), 5.92 (d, J = 8.8 Hz, 1H), 3.04 (s, 3H), 2.41 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.4, 152.7, 142.7, 138.6, 138.0, 134.3, 131.8, 129.9 (q, J = 32.8 Hz), 129.6, 128.8, 128.6 (2 x C), 128.5 (2 x C), 128.2, 128.1, 126.6, 126.5 (q, J = 3.7 Hz, 2 x C), 124.0 (2 x C), 123.9 (q, J = 271 Hz), 123.2, 122.7, 107.3, 35.1, 21.6. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). HRMS (ESI) m / z calcd for C 27 H 23 ON3F3 + [M+H] + 462.1788, found 462.1797.

[0195] Synthesis of compound C12:

[0196] The synthesis step was referred to Example 21, the aldehyde of the first step reaction was selected as m-tolualdehyde, to obtain compound C11, yield: 92%; 1H NMR (400 MHz, CDC13) δ: 7.69 (d, J = 8.4 Hz, 2H), 7.63 (d, J = 8.4 Hz, 2H), 7.58 (d, J = 6.4 Hz, 1H), 7.28-7.25 (m, 6H), 6.97-6.94 (m, 2H), 6.76 (s, 1H), 6.13 (d, J = 8.8 Hz, 1H), 5.95 (d, J = 8.8 Hz, 1H), 3.06 (s, 3H), 2.51 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.4, 153.0, 142.7, 137.0, 136.4, 134.2, 131.6, 131.2, 129.8 (q, J = 32.8 Hz), 129.4, 128.7 (2 x C), 128.6, 128.5 (2 x C), 128.2, 128.1, 126.4 (q, J = 3.7 Hz, 2 x C), 126.1, 123.9 (q, J = 271 Hz), 123.8 (2 x C), 122.9, 110.4, 35.1, 21.4. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). HRMS (ESI) m / z calcd for C 27 H 23 ON3F3 + [M+H] + 462.1788, found 462.1788.

[0197] Synthesis of compound C13 of example 33:

[0198] The synthesis step refers to example 21, the aldehyde of the first step reaction is selected to be trifluoromethylbenzaldehyde to obtain compound C13, yield: 81%; 1 H NMR (400 MHz, CDC13) δ: 7.69 (d, J = 8.4 Hz, 2H), 7.63 (d, J = 8.4 Hz, 2H), 7.58 (d, J = 6.4 Hz, 1H), 7.28-7.25 (m, 6H), 6.97-6.94 (m, 2H), 6.76 (s, 1H), 6.13 (d, J = 8.8 Hz, 1H), 5.95 (d, J = 8.8 Hz, 1H), 3.06 (s, 3H), 2.51 (s, 3H). 13C NMR (100 MHz, CDC13) δ: 161.9, 151.1, 142.5, 138.5, 135.4, 134.1, 130.5 (q, J = 32.2 Hz), 130.3 (q, J = 32.8 Hz), 128.7 (2 x C), 128.5 (2 x C), 128.3, 127.8, 126.6 (q, J = 3.6 Hz, 2 x C), 126.2 (2 x C), 125.9 (q, J = 3.7 Hz, 2 x C), 124.2 (q, J = 270 Hz), 124.0 (2 x C), 123.8 (q, J = 271 Hz), 123.5, 107.5, 35.1. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F), -62.6 (3 x F). HRMS (ESI) m / z calcd for C 27 H 20 ON3F6 + [M+H] + 516.1505, found 516.1496.

[0199] Synthesis of compound C14 of example 34:

[0200] The synthesis step refers to example 21, the aldehyde of the first step reaction is selected as m-trifluoromethylbenzaldehyde to obtain compound C14, yield: 85%; 1 H NMR (400 MHz, CDC13) δ: 8.06 (s, 1H), 7.97 (d, J = 7.6 Hz, 1H), 7.71 (d, J = 8.4 Hz, 2H), 7.63 (d, J = 8.4 Hz, 2H), 7.63-7.61 (m, 1H), 7.54 (t, J = 7.6 Hz, 1H), 7.26-7.24 (m, 3H), 6.92-6.90 (m, 2H), 6.78 (s, 1H), 6.02 (d, J = 8.8 Hz, 1H), 5.94 (d, J = 8.8 Hz, 1H), 3.05 (s, 3H). 13C NMR (100 MHz, CDC13) δ: 162.0, 151.1, 142.5, 138.5, 134.2, 132.8, 131.4 (q, J = 32.2 Hz), 130.2 (q, J = 32.8 Hz), 129.4, 129.2, 128.7 (2 x C), 128.5 (2 x C), 128.3, 127.7, 126.9 (q, J = 271 Hz), 126.6 (q, J = 3.6 Hz, 2 x C), 125.3 (q, J = 3.6 Hz), 124.0 (2 x C), 123.8 (q, J = 271 Hz), 123.6, 122.8 (q, J = 3.4 Hz), 107.2, 35.1. 19 F NMR (376 MHz, CDC13) δ: -62.5 (3 x F), -62.7 (3 x F). HRMS (ESI) m / z calcd for C 27 H 20 ON3F6 + [M+H] + 516.1505, found 516.1509.

[0201] Synthesis of compound C15 of example 35:

[0202] Synthesis step refer to example 21, the aldehyde of the first step reaction is selected as o-trifluoromethylbenzaldehyde, to obtain compound C15, yield: 80%; 1 H NMR (400 MHz, CDC13) δ: 7.78 (d, J = 7.6 Hz, 1H), 7.72-7.66 (m, 3H), 7.63-7.56 (m, 3H), 7.52 (t, J = 7.6 Hz, 1H), 7.24-7.22 (m, 3H), 6.95-6.92 (m, 2H), 6.85 (s, 1H), 6.17 (d, J = 8.8 Hz, 1H), 6.00 (d, J = 8.8 Hz, 1H), 3.06 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.0, 150.6, 142.5, 137.1, 134.2, 132.0, 131.9, 131.6, 130.0 (q, J = 32.8 Hz), 129.6 (q, J = 32.8 Hz), 128.8, 128.6 (2 x C), 128.5 (2 x C), 128.2, 128.1, 126.5 (q, J = 5.5 Hz), 126.4 (q, J = 3.6 Hz, 2 x C), 124.2 (2 x C), 124.1 (q, J = 272 Hz), 123.8 (q, J = 271 Hz), 123.6, 110.9 (q, J = 3.2 Hz), 35.1.19 F NMR (376 MHz, CDCl3) δ: -57.7 (3×F), -62.4 (3×F). HRMS (ESI) m / z calculated value is C 27 H 20 ON3F6 + [M+H] + 516.1505, measured value is 516.1506.

[0203] Example 36 Synthesis of Compound C16:

[0204] The synthesis steps were similar to those in Example 21. Benzaldehyde was selected as the aldehyde in the first step to obtain compound C16 with a yield of 92%. 1 H NMR(400MHz, CDCl3)δ:7.85-7.83(m,2H),7.70(d,J=8.4Hz,2H),7.64(d,J=8.4Hz,2H),7.45-7.42(m,2H),7.39-7.35 (m,1H),7.26-7.24(m,3H),6.92-6.90(m,2H),6.85(s,1H),6.10(d,J=8.8Hz,1H),5.92(d,J=8.8Hz,1H),3.04(s,3H). 13 C NMR (100MHz, CDCl3) δ: 162.3, 152.6, 142.7, 138.1, 134.2, 131.9, 129.9 (q, J = 32.8Hz), 128.9 (2×C), 128.8, 128.6 (2×C) ,128.5(2×C),128.1,128.0,126.5(q,J=3.6Hz,2×C),126.0(2×C),124.0(q,J=271Hz),123.9(2×C),122.7,107.3,35.1. 19 F NMR (376 MHz, CDCl3) δ: -62.4 (3×F). HRMS (ESI) m / z calculated value is C 26 H 21 ON3F3 + [M+H] + 448.1631, measured value is 448.1628.

[0205] Example 37 Synthesis of Compound C17:

[0206] The synthesis steps were similar to those in Example 21. 1-naphthaldehyde was selected as the aldehyde in the first step to obtain compound C17 in 86% yield. 1H NMR (400 MHz, CDC13) δ: 8.49-8.46 (m, 1H), 7.92-7.89 (m, 2H), 7.74-7.68 (m, 5H), 7.56-7.52 (m, 3H), 7.28-7.24 (m, 3H), 6.99-6.97 (m, 2H), 6.89 (s, 1H), 6.16 (d, J = 8.8 Hz, 1H), 5.98 (d, J = 8.8 Hz, 1H), 3.09 (s, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.3, 152.5, 142.7, 137.3, 134.2, 134.1, 131.2, 130.0 (q, J = 32.7 Hz), 129.8, 129.3, 128.7 (2 x C), 128.6, 128.5 (2 x C), 128.3, 128.0, 127.6, 126.8, 126.5 (q, J = 3.6 Hz, 2 x C), 126.1, 125.8, 125.4, 124.0 (2 x C), 123.9 (q, J = 271 Hz), 123.2, 111.1, 35.1. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). HRMS (ESI) m / z calcd for C 30 H 23 ON3F3 + [M+H] + 498.1788, found 498.1785.

[0207] Synthesis of compound C18 of example 38:

[0208] Synthesis step refer to example 21, the first step reaction aldehyde is selected as cyclohexylaldehyde, to obtain compound C18, yield: 82%; 1 H NMR (400 MHz, CDC13) δ: 7.64 (d, J = 8.4 Hz, 2H), 7.53 (d, J = 8.4 Hz, 2H), 7.25-7.22 (m, 3H), 6.90-6.88 (m, 2H), 6.37 (s, 1H), 6.03 (d, J = 8.8 Hz, 1H), 5.87 (d, J = 8.8 Hz, 1H), 2.99 (s, 3H), 2.71-2.65 (m, 1H), 2.07-1.95 (m, 2H), 1.86-1.80 (m, 2H), 1.74-1.70 (m, 1H), 1.46-1.32 (m, 4H), 1.29-1.21 (m, 1H). 13C NMR (100 MHz, CDC13) δ: 162.9, 159.7, 142.8, 136.9, 134.4, 129.5 (q, J = 32.6 Hz), 128.6 (2 x C), 128.5 (2 x C), 128.2, 128.1, 126.4 (q, J = 3.6 Hz, 2 x C), 123.9 (q, J = 271 Hz), 123.8 (2 x C), 122.0, 107.6, 37.6, 35.0, 33.1 (2 x C), 26.3 (2 x C), 26.1. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). HRMS (ESI) m / z calcd for C 26 H 27 ON3F3 + [M+H] + 454.2101, found 454.2101.

[0209] Synthesis of compound C19

[0210] Synthesis step, refer to example 21, the first step of the reaction of aldehyde is selected as n-propyl aldehyde, to obtain compound C19, yield: 82%; 1 H NMR (400 MHz, CDC13) δ: 7.65 (d, J = 8.4 Hz, 2H), 7.54 (d, J = 8.4 Hz, 2H), 7.25-7.22 (m, 3H), 6.89-6.87 (m, 2H), 6.39 (s, 1H), 6.04 (d, J = 8.8 Hz, 1H), 5.87 (d, J = 8.8 Hz, 1H), 2.99 (s, 3H), 2.69 (q, J = 7.6 Hz, 2H), 1.26 (t, J = 7.6 Hz, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.8, 156.2, 142.7, 137.2, 134.3, 129.6 (q, J = 32.7 Hz), 128.6 (2 x C), 128.5 (2 x C), 128.1, 128.0, 126.5 (q, J = 3.6 Hz, 2 x C), 123.9 (q, J = 271 Hz), 123.8 (2 x C), 122.1, 108.7, 35.0, 21.5, 13.7. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). HRMS (ESI) m / z calcd for C 22 H 21 ON3F3 + [M+H] + 400.1631, found 400.1636.

[0211] Synthesis of compound C20:

[0212] The synthesis procedure was referred to example 21, the aldehyde of the first step reaction was selected as n-butyraldehyde, to give compound C20 in yield of 90%; 1 H NMR (400 MHz, CDC13) δ: 7.65 (d, J = 8.4 Hz, 2H), 7.53 (d, J = 8.4 Hz, 2H), 7.25-7.22 (m, 3H), 6.90-6.88 (m, 2H), 6.38 (s, 1H), 6.04 (d, J = 8.8 Hz, 1H), 5.88 (d, J = 8.8 Hz, 1H), 2.99 (s, 3H), 2.64 (t, J = 7.6 Hz, 2H), 1.73-1.64 (m, 2H), 0.98 (t, J = 7.2 Hz, 3H). 13 C NMR (100 MHz, CDC13) δ: 162.8, 154.8, 142.7, 137.1, 134.3, 129.6 (q, J = 32.7 Hz), 128.6 (2 x C), 128.5 (2 x C), 128.2, 128.1, 126.4 (q, J = 3.6 Hz, 2 x C), 123.9 (q, J = 270 Hz), 123.8 (2 x C), 122.2, 109.2, 35.0, 30.2, 22.8, 14.0. 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). HRMS (ESI) m / z calcd for C 23 H 23 ON3F3 + [M + H] + 414.1788, found 414.1780.

[0213] Synthesis of compound C21:

[0214] The synthesis procedure was referred to example 21, the aldehyde of the first step reaction was selected as isobutyraldehyde, to give compound C21 in yield of 82%; 1 H NMR (400 MHz, CDC13) δ: 7.65 (d, J = 8.4 Hz, 2H), 7.54 (d, J = 8.4 Hz, 2H), 7.25-7.22 (m, 3H), 6.89-6.87 (m, 2H), 6.37 (s, 1H), 6.01 (d, J = 8.8 Hz, 1H), 5.87 (d, J = 8.8 Hz, 1H), 3.02 (hept, J = 6.8 Hz, 1H), 2.99 (s, 3H), 1.27 (d, J = 6.8 Hz, 6H). 13C NMR (100 MHz, CDC13) δ: 162.8, 160.5, 142.8, 137.1, 134.4, 129.5 (q, J = 32.7 Hz), 128.6 (2 x C), 128.5 (2 x C), 128.1 (2 x C), 126.4 (q, J = 3.6 Hz, 2 x C), 123.9 (q, J = 270 Hz), 123.8 (2 x C), 122.1, 107.3, 35.0, 28.0, 22.7 (2 x C). 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). HRMS (ESI) m / z calcd for C 23 H 23 ON3F3 + [M+H] + 414.1788, found 414.1785.

[0215] Synthesis of compound C22 of example 42:

[0216] Synthesis step refer to example 21, the aldehyde of the first step reaction is selected as 3,3-dimethylbutanal, to obtain compound C22, yield: 91 %; 1 H NMR (400 MHz, CDC13) δ: 7.64 (d, J = 8.4 Hz, 2H), 7.52 (d, J = 8.4 Hz, 2H), 7.26-7.22 (m, 3H), 6.93-6.90 (m, 2H), 6.40 (s, 1H), 6.06 (d, J = 8.8 Hz, 1H), 5.89 (d, J = 8.8 Hz, 1H), 3.00 (s, 3H), 2.55 (s, 2H), 0.96 (s, 9H). 13 C NMR (100 MHz, CDC13) δ: 162.7, 152.4, 142.7, 136.7, 134.3, 129.6 (q, J = 32.7 Hz), 128.6 (2 x C), 128.5 (2 x C), 128.2, 128.1, 126.4 (q, J = 3.6 Hz, 2 x C), 124.0 (q, J = 270 Hz), 123.9 (2 x C), 122.3, 110.9, 42.2, 35.1, 31.3, 29.6 (3 x C). 19 F NMR (376 MHz, CDC13) δ: -62.4 (3 x F). HRMS (ESI) m / z calcd for C 25 H 27 ON3F3 + [M+H] + 442.2011, found 442.2107.

[0217] Active Examples

[0218] Active Example 1 Screening of the cytotoxic effect and anti-inflammatory activity of cis-enamido-pyrazole derivatives.

[0219] 1. Cell culture

[0220] Mouse monocyte macrophage Raw 264.7 was cultured in DMEM high glucose medium (Gibco, Cat No: 11995065) with 10% (V / V) FBS (Gibco, Cat No: 10270-106), 100 U / mL penicillin and 100 μg / mL streptomycin (Gibco, Cat No: 15140122). The culture condition was 37°C, 5% CO2, and the cells were subcultured when they reached 80% confluence.

[0221] 2. Cytotoxicity detection

[0222] Mouse monocyte macrophage Raw 264.7 was selected and seeded in a 96-well plate at a density of 2 x 10 4 / mL, 100 μL / well. When the cell coverage was about 80%, all synthetic compounds (10 μM) were added, with 3 replicate wells in each group, and a blank control group was set. After 24 h of continuous culture, 10 μL of CCK8 solution was added to each well, and the absorbance OD value was measured at 450 nm wavelength on a microplate reader after 2 h of incubation in a 37°C, 5% CO2 incubator. The cell inhibition rate was calculated.

[0223] 3. Detection of the expression level of NO by the compound

[0224] Raw 264.7 cells were seeded in a 12-well plate at a density of 2 x 10 5 / mL, 1 mL / well. When the cell coverage was about 80%, positive control drugs (dexamethasone, final concentration 1 μM) and candidate compounds (DMSO solvent control, candidate compound 10 μM) were added. After 1 h, LPS was added at a final concentration of 100 ng / mL. LPS was continuously stimulated for 24 h, and the culture supernatant was collected. The standard concentration of Biyun Tian NO detection kit (Cat No: S0021S) was 0, 1, 2, 5, 10, 20, 40, 60, 100 μM, and 50 μL / well was added to the 96-well plate. Then, Griess Reagent I and Griess Reagent II were added to each well at 50 μL each after being restored to room temperature, and the absorbance was measured at 540 nm after mixing well. The concentration of nitric oxide in the sample was calculated according to the standard curve.

[0225] 4. Detection of the expression level of inflammatory factors by the compound

[0226] Raw 264.7 cells were seeded in a 12-well plate at a density of 2 x 105 / mL, 1 mL / well, and when the cell coverage was about 80%, positive control drug (dexamethasone, final concentration 1 μM) and candidate compound (DMSO solvent control, candidate compound 10 μM) were added, and 1 h later, LPS was added at a final concentration of 100 ng / mL. LPS continued to stimulate for 4 h.

[0227] (1) Extraction of RNA and determination of qPCR

[0228] After the cultured cells were completely discarded, 0.5 mL of RA2 in the RNA extraction kit was added to each well for lysis; the cell lysate was collected to extract Total RNA according to the kit instructions (Vazyme, item number: RC112), and the concentration was measured by ultramicro UV-visible spectrophotometer;

[0229] (2) Reverse transcription reaction

[0230] A 20 μL reverse transcription system was established, including 5x Buffer 4 μL, Total RNA X μL (500 ng), DEPC-H2O (16-X) μL, which was placed in an 8-tube tube, labeled with a serial number, and placed in a PCR instrument to set 50℃, 15 min; 85℃, 5 s; 16℃, a time of reverse transcription;

[0231] (3) Fluorescence quantification

[0232] 80 μL of DEPC-H2O was added to the transcribed cDNA to 100 μL, centrifuged and mixed, and stored at -20℃; according to (SYBR 10 μL + DEPC-H2O 10 μL + primer 0.5 μL) multiplied by the number of samples, 18 μL / well was taken to a 96-well plate, 2 μL of cDNA was added, and a 20 μL system was formed, 1200 rpm, 1 min centrifugation and mixing; the sample plate was placed in the CFX Connect Real-Time System (real-time fluorescence quantitative PCR instrument) for detection, and the program was detected according to (95℃, 2 min; 95℃, 20 s; 57℃, 20 s; 72℃, 20 s) for 39 cycles, 95℃, 1 min; 55℃, 30 s; 95℃, 30 s;

[0233] (4) Analysis

[0234] The experimental results were analyzed by 2 -ΔΔCt method, and the calculation formula was as follows: △Ct target gene = Ct target gene - Ct internal reference gene, △△Ct target gene = △Ct experimental group target gene - △Ct control group target gene.2 -ΔΔCt indicates the expression fold of the experimental group

[0235] relative to the control group target gene.

[0236] The primers are shown in Table 1 below

[0237] Table 1 Primer sequences.

[0238]

[0239] The results are as follows Figure 1 As shown, the synthesized cis-enamino-pyrazole derivatives, except A5, did not show obvious toxicity. The inflammation screening results showed that most of the compounds could effectively inhibit the level of NO in Raw264.7 cells, and in addition, could significantly inhibit the secretion level of inflammatory factors IL-1β and IL-6, among which the in vitro anti-inflammatory activity of compound A2 was the strongest.

[0240] Example 2 Study on the inhibition of inflammatory factors in the Raw 264.7 cell model by the cis-enamino-pyrazole derivative (compound A2).

[0241] The cis-enamino-pyrazole derivative (compound A2) was subjected to an inhibition test of the expression level of cell inflammatory factors, and the test method used a conventional ELISA method.

[0242] 1. Cell culture

[0243] The mouse monocyte macrophage Raw 264.7 was cultured using DMEM high-sugar medium (Gibco, catalog number: 11995065) added with 10% (V / V) FBS (Gibco, catalog number: 10270-106), 100 U / mL penicillin and 100 μg / mL streptomycin (Gibco, catalog number: 15140122). The culture conditions were 37°C, 5% CO2, and the cells were subcultured when they reached 80% confluence.

[0244] 2. Cell dosing and cell inflammation induction

[0245] Cells in the logarithmic growth phase were used for the experiment, and the cells were seeded at 2×10 5 / well in a 12-well plate and cultured at 37°C in an environment with 5% CO2 until they grew to 70%, and were ready for use. Groups were set: blank stimulation control group, LPS stimulation group and dosing group (compound A2). The culture medium was carefully removed, and fresh complete medium containing the compound was added to the positive control group and the dosing group, and the same volume of DMSO was added to the blank stimulation control group and the LPS stimulation group. After 1 hour, 1 μg / mL LPS was added to each group except the blank stimulation control group to induce cell inflammation for 4 hours. The well plate was taken out, and the ELISA experiment was performed according to the kit instructions.

[0246] Collecting culture supernatant: collect the supernatant into a 1.5 mL EP tube, centrifuge at 1000 rpm for 10 minutes, and take the supernatant for ELISA detection of cytokines.

[0247] The secretion of mouse IL-6 and TNF-α in the culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA) kit (Bi Yun Tian, item number: IL-6: PI236; TNF-α: PT512). According to the operation steps of the kit, the specific steps are as follows:

[0248] (1) Reagent preparation

[0249] ① Take out from the refrigerator and place at room temperature for 20 minutes. ② Dilute the washing solution (20x) with double distilled water to 1x to prepare the required washing solution. ③ According to the volume marked on the standard label, add the standard diluent to 1 bottle of standard, and incubate at room temperature for 15 minutes. ④ Take 5 clean 1.5 mL centrifuge tubes, and add 250 μL of standard diluent to each tube for standard dilution. Finally, 1000, 500, 250, 125, 62.5, 31.25 pg / mL of six standard concentrations are obtained, and the diluted standard is added to the pre-coated plate hole in turn. Add the standard diluent directly as 0 pg / mL concentration, a total of seven standard concentrations. ⑤ Add 300 μL to each well, and the next one is performed after about 15-30 seconds. A total of five times of plate washing, and dry with force on paper.

[0250] (2) Operation steps

[0251] ① Calculate the number of pre-coated plates required for one experiment, and take the required plates and place them in the 96-well frame.

[0252] ② Add 100 μL / well of sample or different concentrations of standard to the corresponding wells, respectively, and seal the reaction wells with sealing film (transparent). Incubate at room temperature for 120 minutes.

[0253] ③ Wash the plate 5 times, and the last time is placed on thick absorbent paper to dry.

[0254] ④ Add 100 μL / well of biotinylated antibody, and seal the reaction wells with sealing film (transparent). Incubate at room temperature for 60 minutes.

[0255] ⑤ Wash the plate 5 times, and the last time is placed on thick absorbent paper to dry.

[0256] ⑥ Add 100 μL / well of horseradish peroxidase-labeled Streptavidin. Seal the reaction wells with sealing film (white), and incubate at room temperature in the dark for 20 minutes.

[0257] ⑦ Wash the plate 5 times, and the last time is placed on thick absorbent paper to dry.

[0258] 8. Add 100 μL / well of color developing agent TMB solution, cover the reaction wells with sealing film (white), and incubate at room temperature for 20 minutes in the dark.

[0259] 9. Add 50 μL / well of termination solution, mix well, and immediately measure the expression levels of TNF-α and IL-6.

[0260] The results are shown in Table 1. Figure 2 As compared with the LPS group, compound A2 significantly down-regulated the IL-6 inflammatory factor level (IC 50 = 4.816 μM) at concentrations of 10 μM, 20 μM, 50 μM, and 100 μM, down-regulated the IL-1β inflammatory factor level (IC 50 = 90.11 μM) at concentrations of 50 μM and 100 μM, and showed a dose-dependent down-regulation.

[0261] Anti-inflammatory effect of active example 3 compound A2 on PM2.5-induced lung injury in mice

[0262] I. Purpose of the experiment

[0263] To investigate the therapeutic effect of compound A2 on PM2.5-induced lung injury in mice.

[0264] II. Experimental materials

[0265] Experimental animals: 36 C57BL / 6 male mice, 6-8 weeks old, weighing about 18-22 g, purchased from Zhuhai Baisitong Biotechnology Co., Ltd.

[0266] Experimental instruments: heating stirrer, paraffin microtome, microscope, syringe, cell counter, balance, oven, refrigerated centrifuge, blood analyzer, etc.

[0267] Experimental reagents: HE staining related reagents, PBS solution, Masson staining kit (Solabio, catalog number G1340), immunohistochemical kit (Zhongshanjingqiao, catalog number PV9005), DAB color developing kit (Zhongshanjingqiao, catalog number ZLI9018), etc.

[0268] III. Animal experiment

[0269] 1. Establishment of a PM2.5-induced lung injury mouse model: ① Molding drug-PM2.5 particles; ② Molding method-give the mice intratracheal instillation of PM2.5 particle suspension 50 μL at a dose of 3 mg / kg on the first and third days of molding.

[0270] 2. Animal grouping: blank group, model group, positive drug group (dexamethasone), treatment group (compound A2 low, medium and high doses: 15 mg / kg, 30 mg / kg, 60 mg / kg), 6 in each group; body weight was detected every day, see Figure 3 .

[0271] 3. Animal administration: low (15 mg / kg), medium (30 mg / kg), high (60 mg / kg) doses of compound A2 were used for gavage, dexamethasone was selected as the positive control group, the control group and the model group were given the same volume of normal saline for gavage, once a day, for 7 consecutive days, PM2.5 particles were instilled through the trachea on the 1st and 3rd days, 24 hours later, the eyeball was taken for blood, and an overdose of 3% sodium pentobarbital was injected intraperitoneally for euthanasia.

[0272] 4. Animal sampling: after the mice were sacrificed, the lung alveolar lavage fluid and lung tissue were taken.

[0273] 5. Index observation: total number of inflammatory cells, mouse lung tissue pathology, collagen formation, inflammatory protein expression, etc.

[0274] (1) The total number of cells in the mouse lung alveolar lavage fluid was calculated using a cell counter; the total number of white blood cells in the mouse whole blood was counted using a blood analyzer, and the results are shown in Figure 4 , relative to the blank group, the total number of cells in the lung alveolar lavage fluid and the total number of white blood cells in the whole blood of the model group were significantly increased; relative to the model group, dexamethasone and high-dose compound A2 reduced the total number of cells in the lung alveolar lavage fluid and the total number of white blood cells in the whole blood.

[0275] (2) Tissue fixation, dehydration and embedding

[0276] ① The tissues of each group of mice were placed in 10% neutral formaldehyde solution for 24h.

[0277] ② The trimmed tissue block was placed in an embedding box and washed with running water for 24h to completely remove residual formaldehyde.

[0278] ③ The tissue was dehydrated by alcohol gradient, the specific steps were as follows: 70% alcohol for 12h, 80% alcohol for 1.5h, 95% alcohol I for 45min, 95% alcohol II for 30min, 100% alcohol I for 25min, 100% alcohol II for 20min.

[0279] ④ After dehydration, the tissue was placed in an alcohol / dimethylbenzene (1:1, v / v) solution for 20min.

[0280] ⑤ The tissue was immersed in dimethylbenzene I for 20min and then in dimethylbenzene II for 10min.

[0281] ⑥ The tissue was placed in preheated melted paraffin I and paraffin II at 60-65℃ for 1h each.

[0282] ⑦ Pour a small amount of embedding paraffin into the preheated metal embedding frame, place the skin tissue block in it, perpendicular to the embedding frame, with the bottom flat, pour paraffin again, embed the tissue, and cool it.

[0283] ⑧ Paraffin Sectioning: Mount the tissue paraffin block on a Leica microtome and slice serially at a thickness of 5 μm. Using toothless forceps, place the sections in 40°C water for spreading. After scooping the sections with a glass slide, bake them at 60°C for 2 hours and store them in a sectioning box at room temperature until ready for use.

[0284] (3) HE staining

[0285] ① Dewaxing and hydration: Place skin tissue paraffin sections in xylene I and xylene II for 15 minutes each, then place them in 100% ethanol I and ethanol II for 3 minutes each, 95% ethanol I and ethanol II for 3 minutes each, 80% ethanol for 3 minutes, and double-distilled water for 1 minute;

[0286] ②Hematoxylin staining for 15 minutes, then wash away excess stain on the slides;

[0287] ③1% hydrochloric acid ethanol (99 mL 70% ethanol + 1 mL concentrated hydrochloric acid) color separation for 3 seconds, the cell nucleus and chromatin should be clear under the microscope;

[0288] ④ Rinse with running water for 15 minutes to return to blue, and then with distilled water for 1 minute;

[0289] ⑤ Incubate with eosin for 2 minutes, then rinse with running water for 1 minute;

[0290] ⑥ Dehydrate with 80% and 100% ethanol for 2 seconds and 7 minutes respectively;

[0291] ⑦ Xylene I and II for 5 min each;

[0292] ⑧Seal the slide: Take the slide out of xylene II, add neutral gum to the tissue, gently cover with a coverslip, and let it dry naturally;

[0293] ⑨Observation: Observe pathological changes under a microscope, take photos and analyze.

[0294] like Figure 5 As shown, in the model group, alveoli collapsed, alveolar walls thickened, PM2.5 particles aggregated in the intercellular spaces, and there was obvious inflammatory cell infiltration. In the drug-treated group (Compound A2), inflammatory cells were significantly reduced, and the degree of alveolar collapse and alveolar wall thickening was alleviated, indicating that Compound A2 has an alleviating effect on PM2.5-induced lung damage.

[0295] (3) Masson staining

[0296] ① Paraffin sections were routinely dewaxed into distilled water.

[0297] ②According to the operation of Soler Masson kit instructions, 1:1 mixed reagent A1 and A2 before use to prepare Weigert iron hematoxylin staining solution, drop covering section staining 10 min.

[0298] ③Distilled water wash off excess dyeing liquid, drop acid ethanol differentiation liquid differentiation 5-15s, distilled water wash 30s.

[0299] ④Masson blue solution back blue 3 min, distilled water wash 30s.

[0300] ⑤Lichun Hong pink staining solution staining 10 min.

[0301] ⑥In the above operation process according to the ratio of distilled water: weak acid solution = 2:1 configuration of weak acid working solution, drop weak acid working solution wash 30s.

[0302] ⑦Pour off the excess liquid, drop phosphomolybdic acid solution treatment 1 min. Drop weak acid working solution wash 30s.

[0303] ⑧Pour off the excess liquid, drop aniline blue staining solution staining 1 min. Drop weak acid working solution wash 30s.

[0304] ⑨95% ethanol rapid dehydration 2-3s, anhydrous ethanol dehydration 2 times, each 5s.

[0305] ⑩Xylene transparency 2 times, each 2 min, neutral gum sealing.

[0306] Microscopic observation shows (see Figure 6 ), compared with the blank group, the proportion of blue collagen fibers in the model group increased; compared with the model group, the proportion of collagen fibers in the low, medium and high dose groups of compound A2 decreased to different degrees, indicating that compound A2 had a remission treatment effect on pulmonary fibrosis caused by PM2.5 induced lung injury.

[0307] (4) Immunohistochemical staining

[0308] ①The lung tissue paraffin section was placed in the oven, 60℃ baking piece 1h.

[0309] ②Dewaxing: xylene 10 min three times→ anhydrous ethanol I 5 min→ anhydrous ethanol II 5 min→ 95% ethanol 5 min→ 85% ethanol 5 min→ 75% ethanol 5 min→ ddH2O 5 min.

[0310] ③Antigen retrieval: 400ml antigen retrieval solution (800ml dd H2O + 3g sodium citrate + 400mg citric acid, then make up to 1000ml) was boiled in advance, and the slices were placed in a beaker and boiled for 20min. After cooling, the slices were washed with PBS for 3min each time.

[0311] ④Drop 3% hydrogen peroxide, incubate at room temperature for 5min, wash the slices with PBS for 3min each time.

[0312] ⑤Remove PBS, drop 10% goat serum blocking solution, and incubate at room temperature for 1h.

[0313] ⑥Remove the goat serum, dry the slices and directly drop the primary antibody, incubate in a wet box at 4°C overnight (about 14h).

[0314] ⑦Take out the slices, rewarm for 30min, and wash with PBS for 3min each time.

[0315] ⑧Remove PBS, drop the secondary antibody for immunohistochemistry into each sample, incubate at room temperature for 15min, and wash with PBS for 3min each time.

[0316] ⑨Dry the slices, immediately place them under a microscope, drop DAB staining, and wash with PBS for 5min after color development.

[0317] ⑩Hematoxylin counterstaining for 40s → rinse with running water for 2min → return to blue for 7s → rinse with running water for 10-15min → soak in ddH2O for 5min.

[0318] 95% ethanol (1min) → anhydrous ethanol I (1min) → anhydrous ethanol II (1min) → xylene (1min) → xylene (1min) → xylene (1min) neutral resin mounting.

[0319] The results are shown in Figure 7 Compared with the blank control group, the expression levels of iNOS and LY6G, markers of M1 macrophages and neutrophils, were significantly increased, and the expression level of Arg-1, a marker of M2 macrophages, was lower in the model group. In the drug group (compound A2), the expression levels of iNOS and LY6G were significantly reduced, and the expression of Arg-1 was increased, indicating that compound A2 had an inhibitory effect on the inflammatory infiltration of M1 macrophages and neutrophils in lung tissue.

Claims

1. A cis-enamide-pyrazole derivative, characterized in that: Including the following chemical formula, 、 、 、 、 、 、 、 、 、 。 2. A pharmaceutical composition, characterized in that The invention comprises the cis-enamide-pyrazole derivative according to claim 1 and a pharmaceutically acceptable auxiliary material or excipient.

3. Use of the cis-enamide-pyrazole derivative according to claim 1 in the preparation of an inhibitor of IL-1β.

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