A berberine derivative, a pharmaceutical composition and its application

Abstract

This invention provides a berberine derivative, a pharmaceutical composition, and its application, relating to the field of medicinal chemistry. The main skeleton of the berberine derivative is berberine with a urea group. By connecting different substituents to different sites on the berberine skeleton through the urea group and changing the oxidation state of the C ring, a series of berberine derivatives with high activity against soluble epoxide hydrolase were obtained. The IC50 value of the berberine derivatives inhibiting soluble epoxide hydrolase reaches the nanomolar level, indicating that the berberine derivatives have a highly efficient inhibitory effect on soluble epoxide hydrolase. Thus, by inhibiting soluble epoxide hydrolase, an anti-inflammatory effect can be achieved, which has good application scenarios.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical chemistry, in particular to a berberine derivative, a pharmaceutical composition and application thereof. BACKGROUND

[0002] Inflammation is an evolutionarily conserved defense response of the immune system to protect the host from harmful stimuli. Common clinical conditions such as sepsis, cardiovascular and cerebrovascular diseases, neurodegenerative diseases, gastrointestinal diseases, and diabetes are often induced by infection and local tissue damage, accompanied by local inflammation and elevated levels of pro-inflammatory cytokines. This complex and diverse inflammatory development pattern and its role in various diseases have attracted extensive attention from researchers. Therefore, it is possible to target inflammation-related pathways to assist in the treatment of many inflammatory diseases.

[0003] Existing anti-inflammatory drugs can be divided into steroidal anti-inflammatory drugs (hormones) that act on phospholipase A2 and non-steroidal anti-inflammatory drugs that act on cyclooxygenase (COX-2) and lipoxygenase (LOX-2). Due to their significant efficacy, they have been widely used in clinical practice. Steroidal drugs have stronger anti-inflammatory effects, but also have more potential side effects, such as respiratory diseases such as asthma, and osteoporosis. Non-steroidal anti-inflammatory drugs have fewer side effects overall, but the adverse reactions such as gastrointestinal bleeding caused by their widespread use are also increasing.

[0004] Studies have found that soluble epoxide hydrolase (sEH) plays an important role in the process of inflammation, and has a completely new anti-inflammatory pathway compared to existing drug action pathways, which may be a new direction for the treatment of inflammatory diseases. Arachidonic acid (AA) is a direct precursor of active mediators such as prostaglandins and leukotrienes, which can regulate a series of different inflammatory processes. Epoxide eicosatrienoic acid (EETs) produced by cytochrome P450 enzymes from AA is a beneficial arachidonic acid metabolite with anti-inflammatory and anti-apoptotic effects, while the main metabolic pathway of EETs is to hydrolyze pro-inflammatory factor dihydroxy eicosatrienoic acid (DHETs) through sEH. The decrease in EETs concentration and the increase in DHETs concentration in vivo can induce inflammation, so inhibiting the activity of sEH to increase the concentration of EETs in the body can effectively treat inflammation and related diseases. Current research also shows that sEH is closely related to cardiovascular, central nervous system, and metabolic diseases. Several research groups have designed and synthesized compounds with different skeletons targeting sEH, some of which have entered clinical trials, such as EC5026.

[0005]

[0006] Natural products are an important resource for new drug development. Berberine (BBR), also known as Huanglian, is a typical natural product with rich pharmacological activities, and has a long history of medicinal use in China. BBR has various pharmacological activities such as anti-inflammatory, antibacterial, and antiviral activities. More and more in vitro and in vivo experimental models show that BBR is a multi-effect molecule that can target multiple inflammation-related signal molecules and has multiple anti-inflammatory mechanisms. However, the unclear action target and low bioavailability of BBR seriously limit its development.

[0007] Therefore, there is an urgent need to design and develop a compound with a novel berberine skeleton and an IC50 value of sEH at the nanomolar level. SUMMARY

[0008] The present application provides a berberine derivative, a pharmaceutical composition and an application thereof, to solve the problem of high IC50 value and poor drug property of the existing anti-inflammatory compound.

[0009] In a first aspect, the present application provides a berberine derivative, which has the following structural general formula of formula I or formula II:

[0010]

[0011] In the formula I or formula II, R 1 and R 2 are methoxy, or R 1 and R 2 form a methylenedioxy group by forming a ring therebetween;

[0012] R 3 is selected from any one of C 1-4 alkyl, five- to six-membered cycloalkylmethyl, five- to six-membered oxygen-containing heterocycloalkylmethyl, N-(C 1-4 alkylformyl)piperidyl, N-(C 1-4 alkylformyl)piperidylmethyl, N-(halogenated C 1-4 alkylformyl)piperidyl, N-(halogenated C 1-4 alkylformyl)piperidylmethyl, halogenated C 1-3 alkoxyphenyl, halogenated C 1-3 alkoxybenzyl, methylsulfonylbenzyl, aminosulfonylbenzyl;

[0013] C ring is selected from any one of the following structures:

[0014]

[0015] Preferably, R 1 and R 2 form a methylenedioxy group by forming a ring therebetween.

[0016] Preferably, the R 3 is selected from any one of isopropyl, isobutyl, cyclopentyl, cyclohexyl, oxetanemethyl, tetrahydrofuranmethyl, tetrahydropyranmethyl, N-acetylpiperidinyl, N-propionylpiperidinyl, N-acetylpiperidinylmethyl, N-propionylpiperidinylmethyl, difluoromethoxyphenyl, trifluoromethoxyphenyl, difluoromethoxybenzyl, trifluoromethoxybenzyl, methylsulfonylbenzyl, aminosulfonylbenzyl.

[0017] Preferably, the R 3 is selected from any one of trifluoromethoxybenzyl, difluoromethoxybenzyl.

[0018] Preferably, the C ring is selected from any one of the following structures:

[0019]

[0020] Preferably, the C ring is

[0021] Preferably, the R 1 and R 2 form a methylenedioxy group, or the R 1 , R 2 is methoxy, the R 3 is selected from any one of tetrahydrofuranmethyl, N-acetylpiperidinylmethyl, N-propionylpiperidinylmethyl, trifluoromethoxybenzyl, difluoromethoxybenzyl, methylsulfonylbenzyl, or aminosulfonylbenzyl, and the C ring is selected from

[0022] Preferably, the berberine derivative has the following structure:

[0023]

[0024]

[0025] In a second aspect, the present application provides a berberine derivative pharmaceutical composition, wherein the pharmaceutical composition comprises at least one of the compounds, isotope-labeled compounds, solvates, polymorphs, pharmaceutically acceptable salts, or prodrug compounds of any one of the compounds of the first aspect.

[0026] In a third aspect, the present application provides a use of the berberine derivative of any one of the first aspect or the berberine derivative pharmaceutical composition of the second aspect in the preparation of an anti-inflammatory drug, wherein the anti-inflammatory drug is used for inhibiting inflammation caused by soluble epoxide hydrolase.

[0027] The present application includes the following advantages:

[0028] (1) The embodiment of the present application introduces the berberine skeleton in general formula I and II to obtain a novel soluble epoxide hydrolase inhibitor compound, most of the compounds have an IC50 value of less than 20 nM for soluble epoxide hydrolase, and the lowest IC50 value is about 0.67 nM;

[0029] (2) The berberine derivative prepared in the embodiment of the present application for inhibiting soluble epoxide hydrolase has a main skeleton of berberine with a urea group, different substituents are connected to the berberine skeleton at different positions through the urea group, and the oxidation state of the C ring is changed, so that a series of berberine derivatives with high activity for soluble epoxide hydrolase are obtained, and the IC50 value of the berberine derivative for inhibiting soluble epoxide hydrolase reaches the nanomolar level, indicating that the berberine derivative has a high inhibitory effect on soluble epoxide hydrolase, so that the inhibitory effect on inflammation can be achieved by inhibiting soluble epoxide hydrolase, and the berberine derivative has a good application prospect. DETAILED DESCRIPTION

[0030] The following embodiments are provided to better further understand the present application, and do not limit the content and protection scope of the present application, and do not constitute a limitation, anyone who is inspired by the present application or combines the present application with other prior art features to obtain any product which is the same or similar to the present application falls within the protection scope of the present application.

[0031] If the specific experimental steps or conditions are not indicated in the embodiments, the operation or conditions can be performed according to the conventional experimental steps described in the prior art. The various chemicals used in the present application are commercially available, and the nuclear magnetic resonance and mass spectrometer used to determine the structure of the following compounds are provided by the Beijing Institute of Technology Analysis and Testing Center.

[0032] The present application provides a berberine derivative, a pharmaceutical composition and an application thereof in the first aspect, and the berberine derivative has the following structural general formula I or II:

[0033]

[0034] In the formula I or II, R 1 , R 2 are combined into methylenedioxy or are respectively selected as methoxy;

[0035] R 3 is selected from C 1-4 alkyl, five- to six-membered cycloalkane methyl, five- to six-membered oxygen-containing heterocycloalkane methyl, N-(C 1-4 alkyl formyl) piperidyl, N-(C 1-4 alkyl formyl) piperidyl methyl, N-(halogenated C 1-4alkylformyl)piperidinyl, N-(halogenated C) 1-4 alkylformyl)piperidinylmethyl, halo-C 1-3 Alkoxyphenyl, Halogenated C 1-3 Any one of the following groups: alkoxybenzyl, methanesulfonylbenzyl, or aminosulfonylbenzyl;

[0036] The C-ring can be selected from any of the following structures:

[0037]

[0038] The above C 1-4 Alkyl groups represent straight-chain or branched saturated monovalent hydrocarbon groups having 1 to 4 carbon atoms. For example, C 1-4 Alkyl means a straight-chain or branched alkyl group having 1, 2, 3, or 4 carbon atoms, such as methyl, ethyl, propyl, butyl, isopropyl, isobutyl, sec-butyl, or tert-butyl.

[0039] The above C 1-4 Alkyl formyl group is represented as RC(=O)-, where R is C 1-4 Alkyl group. The alkyl formyl group is, for example, acetyl, propionyl, butyryl, valeryl, or isobutyryl.

[0040] The methyl group of the above five- or six-membered cycloalkanes is represented as RCH2-, where R is C. 5-6 Cycloalkyl, wherein the cycloalkyl group is, for example, cyclopentyl or cyclohexyl.

[0041] The methyl group of the aforementioned five- or six-membered oxygen-containing heterocyclic alkanes is represented as a 5- or 6-membered monocyclic ring and contains at least one, for example, 1, 2, 3 or more O heteroatoms. Five-membered rings include tetrahydrofuranyl and dioxacyclopentenyl; or six-membered rings include tetrahydropyranyl and morpholinyl.

[0042] The above C 1-3 Alkoxyphenyl is represented as ROPh-, where RO is C 1-3 Alkoxy group. The alkoxy group may be attached to the 2, 3, or 4 positions of the phenyl group, and the alkoxy group may be, for example, methoxy, difluoromethoxy, trifluoromethoxy, ethoxy, or propoxy.

[0043] The above C 1-3 In alkoxybenzyl groups, it is represented as ROBn-, where RO is C. 1-3 Alkoxy group. The alkoxy group may be attached to the 2, 3, or 4 positions of the benzyl group, and the alkoxy group may be, for example, methoxy, difluoromethoxy, trifluoromethoxy, ethoxy, or propoxy.

[0044] In the present application, the halogen represents fluorine, chlorine, bromine and / or iodine. Correspondingly, the halo- refers to fluoro-, chloro-, bromo- and / or iodo-. Within the scope of the present application, when an atom, residue, group or moiety is halo-, the atom at the halo- position can be mono-, di- or polysubstituted up to per-substitution with halogen atoms, e.g. haloC 1-4 The alkylcarbonyl group can be trifluoroacetyl, and the halo-methoxy group can be difluoromethoxy, or trifluoromethoxy.

[0045] The present application also provides a berberine derivative pharmaceutical composition, which comprises at least one of the berberine derivative, its isotopically-labeled, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound of any one of the first aspect.

[0046] The present application also provides a use of the berberine derivative of any one of the first aspect or the berberine derivative pharmaceutical composition of the second aspect in the preparation of an anti-inflammatory drug, wherein the anti-inflammatory drug is used for inhibiting inflammation caused by soluble epoxide hydrolase.

[0047] The present application also provides a method for preventing and / or treating inflammation-related diseases, which comprises administering to a patient a prophylactically or therapeutically effective amount of at least one of the compound of Formula I or Formula II, its racemate, stereoisomer, tautomer, isotopically-labeled, solvate, polymorph, pharmaceutically acceptable salt, prodrug compound or the pharmaceutical composition.

[0048] In some alternative embodiments, the patient is a mammal, preferably a human.

[0049] The pharmaceutical composition of the present application also comprises any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, which are known to the art and materials, which are used in the pharmaceutical industry. Except insofar as any conventional media or agent is incompatible with the compound, its use in the pharmaceutical composition is contemplated. Supplementary compounds can also be incorporated into the composition.

[0050] In the present application, various pharmaceutically acceptable acids can form pharmaceutically acceptable salts on the basic nitrogen of the compound. The acid can be an inorganic acid or an organic acid; the inorganic acid is hydrochloric acid, sulfuric acid, hydrobromic acid, hydroiodic acid or phosphoric acid; the organic acid is acetic acid, malonic acid, methanesulfonic acid, succinic acid, p-toluenesulfonic acid, citric acid, maleic acid, fumaric acid, malic acid or citric acid.

[0051] The prodrug compounds in the present specification are administered in a certain undisclosed structure but are metabolized or transformed into the compounds disclosed in the present specification in the human body and play a pharmacological role as a pharmacodynamic component. The conventional method for preparing a prodrug is described in "Design of Prodrugs" (H. Bundgaad, Elsevier, 1985).

[0052] The patient refers to any animal including mammals, and the embodiments of the present application preferably refer to mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates, and most preferably refer to humans.

[0053] The therapeutically effective amount refers to the amount of an active compound or pharmaceutical that elicits the biological or medicinal response in a tissue, system, animal, individual or human being sought by a researcher, veterinarian, medical doctor or other clinician, and includes one or more of the following: (1) preventing the disease: for example, in an individual susceptible to or otherwise at risk of a disease, disorder or condition, to prevent the disease, disorder or condition from occurring; (2) inhibiting the disease: for example, in an individual experiencing or exhibiting the pathology or symptomatology of a disease, disorder or condition, to halt the further development of the pathology and / or symptomatology (i.e., to arrest the disease, disorder or condition); and (3) relieving the disease: for example, in an individual experiencing or exhibiting the pathology or symptomatology of a disease, disorder or condition, to reverse the pathology and / or symptomatology (i.e., to cause regression of the disease, disorder or condition).

[0054] To further understand the technical solutions of the present application by those skilled in the art, the following further describes the berberine derivative, pharmaceutical composition and application thereof provided by the present application in specific embodiments.

[0055] The synthesis method of the compound of the present application is briefly described below. In the synthesis examples listed below, the synthesis of intermediates mainly involves Friedel-Crafts acylation, coupling reaction, ring closure reaction and reduction reaction, etc.

[0056] Specifically, the synthesis method of representative specific compounds of the present application is given below.

[0057] Synthesis Example 1: When the urea group is connected to the C10 position of berberine, the synthesis route of the berberine derivative precursor as shown in general formula I is as follows:

[0058]

[0059] The reaction reagents and conditions involved in the above synthetic route are as follows: a: borane-tetrahydrofuran complex, tetrahydrofuran (THF), 4 h; b: pivaloyl chloride, pyridine, dichloromethane (DCM), 5 h; c: zinc chloride, acetic anhydride, 24 h; d: ethylene glycol, p-toluenesulfonic acid, toluene, 12 h; e: palladium on carbon, hydrogen, methanol, 3 h; f: N-bromosuccinimide (NBS), N,N-dimethylformamide (DMF), 12 h; g: acetic anhydride, pyridine, 4-dimethylaminopyridine (DMAP), DCM, 10 min; h: palladium acetate, Xantphos, potassium phosphate, THF, 3 d; i: ammonium chloride, ethanol / water, 2 d; j: sodium borohydride, methanol, 10 min.

[0060] The synthetic route of compound 4 is as follows:

[0061]

[0062] Dissolve 3,4-dimethoxyphenylacetic acid 1 (10.19 mmol, 2.00 g) in 30 mL of tetrahydrofuran, slowly drop borane-tetrahydrofuran complex (1 mol / L, 20.39 mmol, 20.39 mL) at 0 °C under argon protection, and slowly raise the temperature to room temperature after the drop is completed. After the reaction is completed, quench with sodium hydroxide solution, extract with dichloromethane, and then concentrate and dry by rotary evaporation to obtain 3,4-dimethoxyphenyl ethanol 2. Dissolve 2 in dry dichloromethane, add pyridine (20.39 mmol, 1.61 mL), and drop pivaloyl chloride (20.39 mmol, 2.54 mL) at 0 °C. After the reaction is completed, add saturated ammonium chloride solution to terminate the reaction. Wash the organic phase with saturated ammonium chloride solution, distilled water, saturated sodium bicarbonate solution, and saturated sodium chloride solution, respectively, concentrate by rotary evaporation to obtain colorless transparent liquid 3. Redissolve the obtained liquid in 40 mL of acetic anhydride, add zinc chloride (51.04 mmol, 6.95 g), and start the reaction at room temperature under argon atmosphere. After the reaction is completed, slowly drop distilled water under ice bath, extract with ethyl acetate for 3 times, combine the organic phases, wash with saturated sodium bicarbonate solution, concentrate by rotary evaporation to obtain brown viscous liquid, and obtain compound 4 (1.93 g, total yield 61.5%) by column chromatography (silica gel 200-300 mesh, V(petroleum ether):V(ethyl acetate)=15:1).

[0063] HRMS (ESI) (M+Na) + m / z 331.1514, calcd for C 17 H 24 NaO5 + 331.1516.

[0064] 1H NMR (400 MHz, Chloroform-d) δ 7.27 (s, 1H), 6.77 (s, 1H), 4.29 (t, J = 6.7 Hz, 2H), 3.93 (s, 6H), 3.22 (t, J = 6.7 Hz, 2H), 2.58 (s, 3H), 1.16 (s, 9H).

[0065] 13 C NMR (100 MHz, Chloroform-d) δ 199.30, 178.52, 151.53, 146.87, 133.51, 129.39, 114.97, 113.54, 64.98, 56.18, 55.94, 38.70, 33.74, 29.33, 27.24.

[0066] The synthetic route of compound 10 is as follows:

[0067]

[0068] m-nitrobenzaldehyde 6 (39.70 mmol, 6.00 g) was dissolved in 100 mL of toluene, ethylene glycol (79.41 mmol, 4.34 mL), p-toluenesulfonic acid (1.19 mmol, 0.23 g) was added, and the reaction was started at 120 °C. After the reaction was completed, it was washed with saturated sodium bicarbonate solution, saturated sodium chloride solution, and dried and concentrated to obtain a yellow solid. The yellow solid was dissolved in 100 mL of methanol, and palladium-carbon was added under a hydrogen atmosphere. After the reaction was completed, it was filtered and the methanol was evaporated. After drying, the compound was dissolved in DMF, and NBS (43.67 mmol, 7.77 g) was added under ice bath, and the reaction was carried out at room temperature under argon atmosphere. After the reaction was completed, water was added to terminate the reaction, and it was extracted with ethyl acetate several times. The organic phase was combined and washed with half-saturated sodium chloride solution, and dried and concentrated to obtain a black oily compound. Column chromatography (silica gel 200-300 mesh, V(petroleum ether):V(ethyl acetate)=2:1) was used to obtain a brownish red oily compound. The compound obtained above was dissolved in dry dichloromethane, and pyridine (75.12 mmol, 6.05 mL), DMAP (2.57 mmol, 0.32 g), and acetic anhydride (37.51 mmol, 3.55 mL) were added to the reaction system, respectively, to carry out the reaction. After the reaction was completed, distilled water was added to terminate the reaction. The organic phase was washed with saturated ammonium chloride solution, saturated sodium bicarbonate solution, and saturated sodium chloride solution, respectively. It was concentrated by rotary evaporation and dried to obtain a white solid compound 10 (5.33 g, total yield 46.9%).

[0069] HRMS (ESI) (M+Na) + m / z 307.9893, calcd for C 11 H12 BrNNaO3 + 307.9893.

[0070] 1 H NMR (400 MHz, Chloroform-d) δ 7.63 - 7.52 (m, 2H), 7.49 (d, J = 8.4 Hz, 1H), 7.35 - 7.30 (m, 1H), 6.05 (s, 1H), 4.22 - 3.98 (m, 4H), 2.15 (s, 3H).

[0071] 13 C NMR (100 MHz, Chloroform-d) δ 168.35, 137.42, 137.10, 133.49, 122.01, 118.89, 117.05, 102.34, 65.49, 24.59.

[0072] The synthetic route of compound 13 is as follows:

[0073]

[0074] Compound 4 (2.80 mmol, 0.86 g), 10 (1.40 mmol, 0.40 g) were dissolved in 10 mL of tetrahydrofuran, and palladium acetate (0.07 mmol, 0.016 g), Xantphos (0.14 mmol, 0.081 g) and potassium phosphate (2.80 mmol, 0.61 g) were added. After the reaction system was replaced with argon, it was reacted at 85°C for 48 h. After the reaction was completed, it was filtered, concentrated and dried by rotary evaporation to obtain a brown oil. Column chromatography (silica gel 200-300 mesh, V (petroleum ether): V (ethyl acetate) = 1:1) gave white solid compound 11 (0.58 g, 82.8%).

[0075] HRMS (ESI) (M+Na) + m / z 536.2251, calcd for C 28 H 35 NNaO8 + 536.2255.

[0076] 1H NMR (400 MHz, DMSO-d6) δ 9.96 (s, 1H), 7.65 (t, J = 1.9 Hz, 1H), 7.60 (dt, J = 8.3, 2.0 Hz, 1H), 7.50 (d, J = 1.6 Hz, 1H), 7.12 (dd, J = 8.2, 1.6 Hz, 1H), 6.91 (d, J = 1.5 Hz, 1H), 5.74 (d, J = 1.6 Hz, 1H), 4.38 (s, 2H), 4.19 - 4.12 (m, 2H), 3.95 - 3.83 (m, 4H), 3.82 (dd, J = 4.3, 1.6 Hz, 6H), 3.02 (t, J = 6.7 Hz, 2H), 2.03 (d, J = 1.6 Hz, 3H), 1.09 (d, J = 1.6 Hz, 9H).

[0077] 13 C NMR (100 MHz, DMSO-d6) δ 199.95, 177.76, 168.67, 151.26, 147.04, 138.26, 136.52, 132.44, 132.39, 129.78, 129.30, 119.70, 117.83, 115.63, 113.62, 101.98, 65.13, 64.98, 56.27, 56.00, 44.60, 38.59, 33.03, 27.36, 24.41.

[0078] Compound 11 (1.16 mmol, 0.58 g) was dissolved in 15 mL of 1 M ammonium chloride solution of ethanol / water (3:1) and reacted at 90 °C for 24 h, then the temperature was raised to 110 °C and reacted for another 48 h. After the reaction was completed, it was cooled to room temperature and filtered to obtain yellow solid 12 (0.21 g, 62.4%).

[0079] HRMS (ESI) (M-Cl) + m / z 307.1428, calcd for C 19 H 19 N2O2 + 307.1441.

[0080] 1 H NMR (400 MHz, DMSO-d6) δ 9.55 (s, 1H), 8.82 (s, 1H), 7.98 (d, J = 8.9 Hz, 1H), 7.66 (s, 1H), 7.57 (d, J = 9.0 Hz, 1H), 7.12 (s, 1H), 7.07 (s, 1H), 6.56 (s, 2H), 4.78 (t, J = 6.4 Hz, 2H), 3.92 (s, 3H), 3.86 (s, 3H), 3.19 (t, J = 7.3 Hz, 2H).

[0081] Compound 12 (0.61 mmol, 0.21 g) was dissolved in methanol, and sodium borohydride was added to the system at 45 °C. After the reaction was completed, methanol was evaporated and redissolved in dichloromethane, washed with water and saturated sodium chloride solution, and concentrated by rotary evaporation to obtain a yellow solid. Column chromatography (silica gel 200-300 mesh, V(dichloromethane):V(ethanol):V(ammonia water) = 10:0.1:0.05) gave light yellow solid 13 (compound 13) (0.14 g, 74.6%).

[0082] HRMS (ESI) (M+H) + m / z 311.1754, calcd for C 19 H 23 N2O2 + 311.1754.

[0083] 1 H NMR (400 MHz, Methanol-d4) δ 6.99 (d, J = 8.1 Hz, 1H), 6.89 (s, 1H), 6.72 (s, 1H), 6.63 (dd, J = 8.1, 2.4 Hz, 1H), 6.53 (d, J = 2.3 Hz, 1H), 3.93 (d, J = 15.0 Hz, 1H), 3.84 (d, J = 8.8 Hz, 6H), 3.70 - 3.57 (m, 2H), 3.40 (dd, J = 16.0, 4.2 Hz, 1H), 3.23 - 3.14 (m, 1H), 3.13 - 3.04 (m, 1H), 2.79 - 2.68 (m, 2H), 2.62 (td, J = 11.3, 3.8 Hz, 1H).

[0084] 13 C NMR (175 MHz, Methanol-d4) δ 147.82, 147.71, 145.24, 134.02, 129.54, 129.01, 126.33, 123.66, 114.59, 112.36, 111.52, 108.99, 60.11, 58.22, 55.27, 54.99, 51.18, 34.96, 27.87.

[0085] The synthesis route of compound 5 is as follows:

[0086]

[0087] According to the preparation method of compound 4, compound 5 (2.19 g, total yield 67.5%) can be obtained by borane reduction, acyl chloride protection, and acetylation of compound piperonyl acetic acid (11.10 mmol, 2.00 g).

[0088] HRMS (ESI) (M+Na) + m / z 315.1198, calcd for C 16 H 20 NaO5 + 315.1203.

[0089] 1 H NMR (400 MHz, Chloroform-d) δ 7.23 (s, 1H), 6.75 (s, 1H), 6.02 (s, 2H), 4.26 (t, J = 6.6 Hz, 2H), 3.17 (t, J = 6.6 Hz, 2H), 2.53 (s, 3H), 1.16 (s, 9H).

[0090] The synthetic route of compound 14 is as follows:

[0091]

[0092] According to the preparation method of compound 13, compound 14 (0.15 g, total yield 36.2%) was obtained from compound 5 (2.80 mmol, 0.82 g), 10 (1.40 mmol, 0.40 g) by coupling, ring closing, reduction reaction.

[0093] HRMS (ESI) (M+H) + m / z 295.1438, calcd for C 18 H 19 N2O2 + 295.1441.

[0094] 1 H NMR (400 MHz, Methanol-d4) δ 6.93 (d, J = 8.1 Hz, 1H), 6.79 (s, 1H), 6.63 - 6.55 (m, 2H), 6.49 (d, J = 2.3 Hz, 1H), 5.87 (s, 2H), 3.90 (d, J = 15.0 Hz, 1H), 3.63 (d, J = 15.0 Hz, 1H), 3.55 (dd, J = 11.6, 4.1 Hz, 1H), 3.28 (dd, J = 16.1, 3.7 Hz, 2H), 3.18 - 3.10 (m, 1H), 3.05 (ddd, J = 16.8, 11.6, 5.3 Hz, 1H), 2.74 - 2.62 (m, 2H), 2.58 (td, J = 11.4, 3.8 Hz, 1H).

[0095] 13C NMR (100 MHz, Methanol-d4) δ 146.49, 146.32, 145.32, 133.75, 130.18, 129.05, 126.96, 123.39, 114.62, 112.28, 107.76, 105.06, 100.76, 60.35, 58.05, 50.99, 35.01, 28.26.

[0096] Synthesis of compound C1:

[0097]

[0098] The synthetic route of compound C1 is as follows:

[0099]

[0100] Compound 14 (0.34 mmol, 0.10 g) was dissolved in 10 mL of dichloromethane, 4-dimethylaminopyridine (DMAP, 0.68 mmol, 0.083 g) was added, and finally carbonyldiimidazole (CDI, 0.68 mmol, 0.11 g) was added, and the reaction was carried out at room temperature for 6 h under an argon atmosphere. After the reaction was completed, it was concentrated by rotary evaporation and redissolved in dry DMF, isobutylamine (0.85 mmol, 0.062 g) was added, and stirring was carried out at room temperature for 5 h. After the reaction was completed, distilled water was added to the reaction solution to terminate the reaction, and extraction was carried out twice with ethyl acetate, the organic phases were combined, and washing was carried out with water, saturated ammonium chloride solution, and saturated sodium chloride solution in this order, and the concentrated by rotary evaporation, and column chromatography (silica gel 200-300 mesh, V(dichloromethane):V(ethanol)=10:0.15) was carried out to obtain compound 15 (compound C1) as a light yellow solid (0.075 g, yield 56.2%).

[0101] m.p. 126.7-127.5 °C. HRMS (ESI) (M+H) + m / z 394.2120, calcd for C 23 H 28 N3O3 + 394.2125.

[0102] 1H NMR (400 MHz, Methanol-d4) δ 7.19 (d, J = 2.1 Hz, 1H), 7.15 - 7.04 (m, 2H), 6.81 (s, 1H), 6.59 (s, 1H), 5.89 (s, 2H), 3.97 (d, J = 15.1 Hz, 1H), 3.69 (d, J = 15.1 Hz, 1H), 3.60 (dd, J = 11.5, 4.0 Hz, 1H), 3.41 - 3.32 (m, 1H), 3.17 (ddd, J = 11.1, 5.4, 1.9 Hz, 1H), 3.12 - 2.95 (m, 3H), 2.81 - 2.56 (m, 3H), 1.84 - 1.69 (m, 1H), 0.94 (d, J = 6.7 Hz, 6H).

[0103] 13 C NMR (100 MHz, Methanol-d4) δ 157.04, 146.52, 146.38, 137.74, 133.70, 130.00, 128.76, 127.66, 126.99, 117.58, 116.03, 107.78, 105.06, 100.78, 60.09, 57.98, 50.94, 46.91, 35.12, 28.76, 28.27, 18.99.

[0104] Synthesis of compound C2:

[0105]

[0106] The synthesis route of compound C2 is as follows:

[0107]

[0108] According to the preparation method of compound C1, after reacting with CDI, DMAP to introduce a carbonyl imidazole group, and then reacting with 4-aminomethyl tetrahydro pyran (0.81 mmol, 0.092 g), compound C2 (0.085 g, 58.4%) was obtained as a light yellow solid after purification and drying.

[0109] m.p. 125.4-126.6 °C. HRMS (ESI) (M+H) + m / z 452.2539, calcd for C 26 H 34 N3O4 + 452.2544.

[0110] 1H NMR (400 MHz, Methanol-d4) δ 7.19 (d, J = 2.0 Hz, 1H), 7.15 - 7.06 (m, 2H), 6.87 (s, 1H), 6.70 (s, 1H), 4.04 - 3.91 (m, 3H), 3.81 (d, J = 9.9 Hz, 6H), 3.69 (d, J = 15.1 Hz, 1H), 3.62 (dd, J = 11.5, 4.1 Hz, 1H), 3.48 - 3.35 (m, 3H), 3.18 (ddd, J = 11.0, 5.4, 1.9 Hz, 1H), 3.09 (d, J = 6.7 Hz, 3H), 2.82 - 2.67 (m, 2H), 2.62 (td, J = 11.4, 3.8 Hz, 1H), 1.82 - 1.69 (m, 1H), 1.65 (ddd, J = 13.3, 4.0, 2.0 Hz, 2H), 1.30 (dtd, J = 13.4, 11.8, 4.6 Hz, 2H).

[0111] 13 C NMR (100 MHz, Methanol-d4) δ 157.03, 147.95, 147.80, 137.63, 133.86, 129.27, 128.77, 127.92, 126.32, 117.66, 116.18, 111.63, 109.11, 67.36, 59.82, 58.07, 55.34, 55.04, 51.09, 45.07, 35.67, 35.02, 30.34, 27.84.

[0112] Synthesis of compound C3:

[0113]

[0114] The synthesis route of compound C3 is as follows:

[0115]

[0116] According to the preparation method of compound C1, after reacting with CDI, DMAP to introduce a carbonyl imidazole group, and then reacting with 4-aminomethyl tetrahydro pyran (0.85 mmol, 0.098 g), compound 14 (0.34 mmol, 0.10 g) was dried after purification to obtain light yellow solid 17 (compound C3) (0.08 g, 53.9%).

[0117] m.p. 117.5-118.4 °C. HRMS (ESI) (M+H) + m / z 436.2224, calcd for C 25 H 30 N3O4+ 436.2231.

[0118] 1 H NMR (400 MHz, Methanol-d4) δ 7.18 (d, J = 2.1 Hz, 1H), 7.14 - 7.04 (m, 2H), 6.81 (s, 1H), 6.58 (s, 1H), 5.88 (s, 2H), 4.02 - 3.91 (m, 3H), 3.66 (d, J = 15.0 Hz, 1H), 3.57 (dd, J = 11.8, 4.0 Hz, 1H), 3.45 - 3.32 (m, 3H), 3.19 - 2.99 (m, 4H), 2.79 - 2.53 (m, 3H), 1.82 - 1.68 (m, 1H), 1.68 - 1.59 (m, 2H), 1.37 - 1.22 (m, 2H).

[0119] 13 C NMR (100 MHz, Methanol-d4) δ 157.02, 146.48, 146.33, 137.62, 133.91, 130.16, 128.77, 127.87, 127.08, 117.62, 116.14, 107.77, 105.06, 100.75, 67.36, 60.08, 58.05, 50.98, 45.07, 35.67, 35.21, 30.34, 28.36.

[0120] Synthesis of compound C4:

[0121]

[0122] The synthesis route of compound C4 is as follows:

[0123]

[0124] According to the preparation method of compound C1, after the reaction of compound 13 (0.32 mmol, 0.10 g) with CDI, DMAP to introduce a carbonyl imidazole group, and then with (S)-tetrahydrofurfuryl-2-methylamine (0.81 mmol, 0.081 g), light yellow solid 18 (compound C4) (0.062 g, 43.9%) was obtained after purification and drying.

[0125] m.p. 105.7-106.3 °C. HRMS (ESI) (M+H) + m / z 438.2385, calcd for C 25 H 32 N3O4 + 438.2387.

[0126] 1 H NMR (400 MHz, Methanol-d4) δ 7.18 (d, J = 2.1 Hz, 1H), 7.16 - 7.06 (m, 2H), 6.87 (s, 1H), 6.70 (s, 1H), 4.03 - 3.92 (m, 2H), 3.88 (dt, J = 8.3, 6.7 Hz, 1H), 3.81 (d, J = 9.7 Hz, 6H), 3.78 - 3.70 (m, 1H), 3.70 - 3.62 (m, 1H), 3.58 (dd, J = 11.5, 4.1 Hz, 1H), 3.42 (dd, J = 16.3, 4.1 Hz, 1H), 3.35 (dd, J = 13.8, 4.1 Hz, 1H), 3.26 - 3.12 (m, 2H), 3.08 (ddd, J = 16.6, 11.8, 3.4 Hz, 1H), 2.81 - 2.65 (m, 2H), 2.60 (td, J = 11.3, 3.8 Hz, 1H), 2.08 - 1.84 (m, 3H), 1.69 - 1.56 (m, 1H).

[0127] 13 C NMR (100 MHz, Methanol-d4) δ 156.99, 147.87, 147.74, 137.59, 134.02, 129.37, 128.77, 128.00, 126.36, 117.53, 116.09, 111.56, 109.03, 78.22, 67.70, 59.81, 58.14, 55.30, 55.00, 51.12, 43.22, 35.09, 28.10, 27.90, 25.37.

[0128] Synthesis of compound C5:

[0129]

[0130] The synthetic route of compound C5 is as follows:

[0131]

[0132] According to the preparation method of compound C1, after the reaction of compound 13 (0.32 mmol, 0.10 g) with CDI, DMAP to introduce a carbonyl imidazole group and the reaction with cyclohexylmethylamine (0.81 mmol, 0.091 g), light yellow solid 20 (compound C5) (0.086 g, 59.4%) was obtained after purification and drying.

[0133] m.p. 108.4-109.0 °C. HRMS (ESI) (M+H) +m / z 450.2751, calcd for C 27 H 36 N3O3 + 450.2751.

[0134] 1 H NMR (400 MHz, Methanol-d4) δ 7.21 (d, J = 2.1 Hz, 1H), 7.18 - 7.08 (m, 2H), 6.89 (s, 1H), 6.73 (s, 1H), 4.00 (d, J = 15.1 Hz, 1H), 3.84 (d, J = 9.6 Hz, 6H), 3.72 (d, J = 15.1 Hz, 1H), 3.65 (dd, J = 11.8, 3.8 Hz, 1H), 3.46 (dd, J = 16.3, 4.1 Hz, 1H), 3.21 (ddd, J = 11.1, 5.4, 1.9 Hz, 1H), 3.12 (td, J = 11.4, 5.8 Hz, 1H), 3.05 (d, J = 6.8 Hz, 2H), 2.84 - 2.70 (m, 2H), 2.65 (td, J = 11.4, 3.8 Hz, 1H), 1.83 - 1.74 (m, 4H), 1.76 - 1.66 (m, 1H), 1.48 (dddd, J = 14.8, 11.4, 5.9, 3.5 Hz, 1H), 1.35 - 1.19 (m, 3H), 0.98 (qd, J = 13.6, 12.7, 3.6 Hz, 2H).

[0135] 13 C NMR (100 MHz, Methanol-d4) δ 157.05, 147.92, 147.77, 137.74, 133.75, 129.17, 128.77, 127.74, 126.24, 117.58, 116.05, 111.55, 109.00, 59.82, 58.04, 55.30, 55.01, 51.08, 45.74, 38.41, 34.98, 30.53, 27.80, 26.23, 25.67.

[0136] Synthesis of compound C6:

[0137]

[0138] The synthesis route of compound C6 is as follows:

[0139]

[0140] According to the preparation method of compound C1, compound 13 (0.32 mmol, 0.10 g) reacted with CDI, DMAP to access carbonyl imidazole group, then reacted with 4- aminopropionylpiperidine (0.81 mmol, 0.13 g), after drying by purification, light yellow solid 21 (compound C6) (0.095 g, 60.1%) was obtained.

[0141] m.p. 122.3-122.9 °C. HRMS (ESI) (M+H) + m / z 493.2811, calcd for C 28 H 37 N4O4 + 493.2809.

[0142] 1 H NMR (400 MHz, Methanol-d4) δ 7.18 (d, J = 3.5 Hz, 1H), 7.15 - 7.07 (m, 2H), 6.88 (s, 1H), 6.71 (s, 1H), 4.37 (dt, J = 16.2, 4.4 Hz, 1H), 3.98 (d, J = 15.1 Hz, 1H), 3.94 - 3.86 (m, 1H), 3.82 (d, J = 9.9 Hz, 7H), 3.69 (d, J = 15.1 Hz, 1H), 3.62 (dd, J = 11.4, 4.0 Hz, 1H), 3.44 (dd, J = 16.3, 4.1 Hz, 1H), 3.27 - 3.14 (m, 2H), 3.09 (ddd, J = 16.7, 11.9, 5.5 Hz, 1H), 2.89 (ddd, J = 13.9, 11.4, 3.0 Hz, 1H), 2.82 - 2.67 (m, 2H), 2.63 (td, J = 11.3, 3.8 Hz, 1H), 2.42 (q, J = 7.5 Hz, 2H), 1.98 (dd, J = 30.4, 12.9 Hz, 2H), 1.46 - 1.30 (m, 2H), 1.12 (t, J = 7.5 Hz, 3H).

[0143] 13 C NMR (175 MHz, Methanol-d4) δ 173.32, 156.10, 147.89, 147.74, 137.48, 134.00, 129.32, 128.78, 128.04, 126.34, 117.55, 116.10, 111.55, 109.02, 59.81, 58.12, 55.30, 55.00, 51.12, 46.70, 44.12, 40.40, 35.07, 32.67, 31.78, 27.89, 25.90, 8.61.

[0144] Synthesis of compound C7:

[0145]

[0146] The synthetic route of compound C7 is as follows:

[0147]

[0148] According to the preparation method of compound C1, after the reaction of compound 14 (0.34 mmol, 0.10 g) with CDI, DMAP to introduce a carbonyl imidazole group, and then reacted with 4-aminopropionylpiperidine (0.85 mmol, 0.133 g), after purification and drying, light yellow solid 22 (compound C7) (0.083 g, 51.4%) was obtained.

[0149] m.p. 185.7-187.1 °C. HRMS (ESI) (M+H) + m / z 477.2485, calcd for C 27 H 33 N4O4 + 477.2496.

[0150] 1 H NMR (400 MHz, Methanol-d4) δ 7.17 (d, J = 2.7 Hz, 1H), 7.14 - 7.03 (m, 2H), 6.81 (s, 1H), 6.58 (s, 1H), 5.88 (s, 2H), 4.40 - 4.32 (m, 1H), 3.99 - 3.85 (m, 2H), 3.81 (tt, J = 10.4, 4.1 Hz, 1H), 3.65 (d, J = 15.0 Hz, 1H), 3.56 (dd, J = 11.5, 4.0 Hz, 1H), 3.34 (dd, J = 16.7, 4.4 Hz, 1H), 3.22 (ddd, J = 14.1, 11.4, 2.9 Hz, 1H), 3.14 (ddd, J = 11.1, 5.4, 1.9 Hz, 1H), 3.05 (ddd, J = 16.8, 11.5, 5.4 Hz, 1H), 2.89 (ddd, J = 13.9, 11.3, 3.0 Hz, 1H), 2.79 - 2.63 (m, 2H), 2.58 (td, J = 11.3, 3.7 Hz, 1H), 2.42 (q, J = 7.5 Hz, 2H), 2.05 - 1.89 (m, 2H), 1.47 - 1.26 (m, 2H), 1.12 (t, J = 7.5 Hz, 3H).

[0151] 13C NMR (100 MHz, Methanol-d4) δ 173.33, 156.11, 146.48, 146.32, 137.48, 133.97, 130.20, 128.77, 127.96, 127.10, 117.55, 116.09, 107.77, 105.07, 100.75, 60.07, 58.06, 50.98, 46.70, 44.11, 40.39, 35.23, 32.67, 31.77, 28.38, 25.90, 8.61.

[0152] Synthesis of compound C8:

[0153]

[0154] The synthetic route of compound C8 is as follows:

[0155]

[0156] According to the preparation method of compound C1, after the reaction of compound 13 (0.32 mmol, 0.10 g) with CDI, DMAP to introduce carbonyl imidazole group, and the reaction with 4-aminomethyl propionyl piperidine (0.81 mmol, 0.14 g), after purification and drying, light yellow solid 23 (compound C8) (0.080 g, 49.1%) was obtained.

[0157] m.p. 121.3-122.5 °C. HRMS (ESI) (M+H) + m / z 507.2971, calcd for C 29 H 39 N4O4 + 507.2966.

[0158] 1H NMR (400 MHz, Methanol-d4) δ 7.19 (d, J = 1.9 Hz, 1H), 7.16 - 7.05 (m, 2H), 6.88 (s, 1H), 6.71 (s, 1H), 4.54 (ddt, J = 13.0, 4.5, 2.4 Hz, 1H), 4.02 - 3.94 (m, 2H), 3.82 (d, J = 9.8 Hz, 6H), 3.70 (d, J = 15.1 Hz, 1H), 3.63 (dd, J = 11.6, 4.1 Hz, 1H), 3.45 (dd, J = 16.3, 4.1 Hz, 1H), 3.23 - 3.01 (m, 5H), 2.82 - 2.67 (m, 2H), 2.67 - 2.56 (m, 2H), 2.40 (q, J = 7.5 Hz, 2H), 1.85 - 1.73 (m, 3H), 1.24 - 1.02 (m, 5H).

[0159] 13 C NMR (100 MHz, Methanol-d4) δ 173.25, 157.00, 147.91, 147.76, 137.61, 133.90, 129.26, 128.78, 127.96, 126.31, 117.64, 116.16, 111.56, 109.02, 59.83, 58.09, 55.30, 55.01, 51.11, 45.40, 44.55, 41.54, 36.81, 35.04, 30.08, 29.27, 27.85, 25.99, 8.65.

[0160] Synthesis of compound C9:

[0161]

[0162] The synthetic route of compound C9 is as follows:

[0163]

[0164] According to the preparation method of compound C1, after reacting with CDI, DMAP to introduce carbonyl imidazole group, and then reacting with 4-aminomethyl propionyl piperidine (0.85 mmol, 0.15 g), compound 24 (compound C9) was obtained as a light yellow solid (0.075 g, 44.9%) after purification and drying.

[0165] m.p. 120.7 - 122.1 °C. HRMS (ESI) (M+H) + m / z 491.2650, calcd for C 28 H 35 N4O4+ 491.2653.

[0166] 1 H NMR (400 MHz, Methanol-d4) δ 7.18 (d, J = 2.1 Hz, 1H), 7.15 - 7.05 (m, 2H), 6.81 (s, 1H), 6.59 (s, 1H), 5.89 (s, 2H), 4.54 (ddt, J = 13.3, 4.5, 2.3 Hz, 1H), 4.00 - 3.91 (m, 2H), 3.66 (d, J = 15.1 Hz, 1H), 3.57 (dd, J = 11.2, 4.0 Hz, 1H), 3.36 (dd, J = 16.3, 4.2 Hz, 1H), 3.19 - 2.99 (m, 5H), 2.79 - 2.55 (m, 4H), 2.40 (q, J = 7.5 Hz, 2H), 1.84 - 1.73 (m, 3H), 1.24 - 1.03 (m, 5H).

[0167] 13 C NMR (175 MHz, Methanol-d4) δ 173.24, 156.99, 146.49, 146.33, 137.60, 133.92, 130.15, 128.77, 127.88, 127.08, 117.59, 116.11, 107.77, 105.06, 100.76, 60.09, 58.06, 50.99, 45.39, 44.55, 41.54, 36.80, 35.21, 30.08, 29.27, 28.36, 25.99, 8.65.

[0168] Synthesis of compound C10:

[0169]

[0170] The synthetic route of compound C10 is as follows:

[0171]

[0172] According to the preparation method of compound C1, after reacting with CDI, DMAP to introduce a carbonyl imidazole group, and then reacting with 4-trifluoromethoxybenzylamine (0.81 mmol, 0.15 g), compound 25 (compound C10) was obtained as a light yellow solid (0.073 g, 42.9%) after purification and drying.

[0173] m.p. 112.2-113.1 °C. HRMS (ESI) (M+H) +m / z 528.2106, calcd for C 28 H 29 F3N3O4 + 528.2105.

[0174] 1 H NMR (400 MHz, Methanol-d4) δ 7.45 - 7.37 (m, 2H), 7.26 - 7.17 (m, 3H), 7.17 - 7.06 (m, 2H), 6.86 (s, 1H), 6.69 (s, 1H), 4.39 (s, 2H), 3.95 (d, J = 15.2 Hz, 1H), 3.81 (d, J = 9.8 Hz, 6H), 3.70 - 3.62 (d, J = 15.2 Hz, 1H), 3.58 (dd, J = 11.5, 4.0 Hz, 1H), 3.42 (dd, J = 16.3, 4.1 Hz, 1H), 3.19 - 3.03 (m, 2H), 2.81 - 2.65 (m, 2H), 2.59 (td, J = 11.3, 3.7 Hz, 1H).

[0175] 13 C NMR (100 MHz, Methanol-d4) δ 156.83, 148.08, 147.88, 147.74, 139.18, 137.46, 134.04, 129.34, 128.80, 128.48, 128.18, 126.35, 120.72, 120.55 117.76, 116.36, 111.56, 109.02, 59.78, 58.11, 55.29, 55.00, 51.11, 42.38, 35.08, 27.89.

[0176] Synthesis of compound C11:

[0177]

[0178] The synthetic route of compound C11 is as follows:

[0179]

[0180] According to the preparation method of compound C1, after the reaction of compound 14 (0.34 mmol, 0.10 g) with CDI, DMAP to introduce a carbonyl imidazole group, and the reaction with 4-trifluoromethoxybenzylamine (0.85 mmol, 0.16 g), after purification and drying, yellow solid 26 (compound C11) (0.089 g, 51.1%) was obtained.

[0181] m.p. 168.4-169.8 °C. HRMS (ESI) (M+H)+ m / z 512.1808, calcd for C 27 H 25 F3N3O4 + 512.1792.

[0182] 1 H NMR (400 MHz, Methanol-d4) δ 7.44 (d, J = 8.3 Hz, 2H), 7.31 - 7.20 (m, 3H), 7.19 - 7.08 (m, 2H), 6.84 (s, 1H), 6.61 (s, 1H), 5.91 (s, 2H), 4.42 (s, 2H), 3.99 (d, J = 15.1 Hz, 1H), 3.69 (d, J = 15.1 Hz, 1H), 3.60 (dd, J = 11.1, 4.0 Hz, 1H), 3.39 (dd, J = 16.3, 4.2 Hz, 1H), 3.18 (ddd, J = 11.2, 5.4, 1.9 Hz, 1H), 3.08 (ddd, J = 16.8, 11.6, 5.4 Hz, 1H), 2.86 - 2.56 (m, 3H).

[0183] 13 C NMR (100 MHz, Methanol-d4) δ 156.84, 146.50, 146.35, 139.19, 137.50, 133.88, 130.09, 128.79, 128.48, 128.02, 127.05, 120.72, 117.77, 116.30, 107.77, 105.05, 100.77, 60.09, 58.01, 50.97, 42.37, 35.19, 28.32.

[0184] Synthesis of compound C12:

[0185]

[0186] The synthesis route of compound C12 is as follows:

[0187]

[0188] According to the preparation method of compound C1, after reacting with compound 13 (0.32 mmol, 0.10 g) with CDI, DMAP to introduce a carbonyl imidazole group, and then reacting with 4-trifluoromethoxy aniline (0.81 mmol, 0.14 g), a light yellow solid 27 (compound C12) (0.081 g, 49.0%) was obtained after purification and drying.

[0189] M.p. 110.5-111.3 °C. HRMS (ESI) (M+H) + m / z 514.1945, calcd for C 27 H 27 F3N3O4 + 514.1948.

[0190] 1 H NMR (400 MHz, Methanol-d4) δ 7.55 - 7.46 (m, 2H), 7.27 (d, J = 2.2 Hz, 1H), 7.23 - 7.15 (m, 3H), 7.14 (d, J = 8.3 Hz, 1H), 6.88 (m, 1H), 6.70 (s, 1H), 3.99 (d, J = 15.1 Hz, 1H), 3.82 (d, J = 10.8 Hz, 6H), 3.70 (d, J = 15.1 Hz, 1H), 3.61 (dd, J = 11.5, 4.0 Hz, 1H), 3.45 (dd, J = 16.3, 4.0 Hz, 1H), 3.18 (ddd, J = 11.0, 5.5, 1.9 Hz, 1H), 3.10 (ddd, J = 16.7, 12.2, 3.5 Hz, 1H), 2.83 - 2.67 (m, 2H), 2.62 (td, J = 11.3, 3.7 Hz, 1H).

[0191] 13 C NMR (100 MHz, Methanol-d4) δ 153.81, 147.90, 147.75, 143.94, 138.42, 136.94, 134.14, 129.32, 128.86, 128.66, 126.36, 121.34, 120.62, 119.87, 117.86, 116.49, 111.56, 109.03, 59.78, 58.10, 55.30, 55.00, 51.12, 35.11, 27.90.

[0192] Synthesis of compound C13:

[0193]

[0194] The synthesis route of compound C13 is as follows:

[0195]

[0196] According to the preparation method of compound C1, compound 13 (0.32 mmol, 0.10 g) was reacted with CDI, DMAP to introduce carbonyl imidazole group, then reacted with 3- trifluoromethoxybenzylamine (0.81 mmol, 0.15 g), and after purification and drying, light yellow solid 28 (compound C13) (0.097 g, 57.4%) was obtained.

[0197] m.p. 85.2-86.1 °C. HRMS (ESI) (M+H) + m / z 528.2106, calcd for C 28 H 29 F3N3O4 + 528.2105.

[0198] 1 H NMR (400 MHz, Methanol-d4) δ 7.42 (t, J = 7.9 Hz, 1H), 7.33 (dt, J = 8.0, 1.3 Hz, 1H), 7.24 (s, 1H), 7.20 (d, J = 2.0 Hz, 1H), 7.18 - 7.08 (m, 3H), 6.87 (s, 1H), 6.70 (s, 1H), 4.42 (s, 2H), 3.97 (d, J = 15.1 Hz, 1H), 3.81 (d, J = 9.9 Hz, 6H), 3.68 (d, J = 15.0 Hz, 1H), 3.64 - 3.56 (m, 1H), 3.44 (dd, J = 16.3, 4.1 Hz, 1H), 3.17 (ddd, J = 11.0, 5.5, 1.9 Hz, 1H), 3.09 (ddd, J = 16.8, 12.0, 3.5 Hz, 1H), 2.82 - 2.66 (m, 2H), 2.61 (td, J = 11.4, 3.8 Hz, 1H).

[0199] 13 C NMR (175 MHz, Methanol-d4) δ 156.85, 149.34, 147.88, 147.74, 142.80, 137.42, 134.04, 129.77, 129.35, 128.79, 128.23, 126.35, 125.56, 120.54, 119.30, 119.06, 117.83, 116.42, 111.55, 109.00, 59.82, 58.12, 55.28, 55.00, 51.12, 42.51, 35.09, 27.89.

[0200] Synthesis Example 15 - Synthesis of compound C14:

[0201]

[0202] The synthetic route of compound C14 is as follows:

[0203]

[0204] According to the preparation method of compound C1, after the reaction of compound 14 (0.34 mmol, 0.10 g) with CDI, DMAP to introduce a carbonyl imidazole group, and then with 3-trifluoromethoxybenzylamine (0.85 mmol, 0.16 g), a light yellow solid 29 (compound C14) (0.073 g, 42.2%) was obtained after purification and drying.

[0205] m.p. 149.7-151.5 °C. HRMS (ESI) (M+H) + m / z 512.1790, calcd for C 27 H 25 F3N3O4 + 512.1792.

[0206] 1 H NMR (400 MHz, Methanol-d4) δ 7.42 (t, J = 7.9 Hz, 1H), 7.33 (d, J = 7.7 Hz, 1H), 7.24 (s, 1H), 7.20 (s, 1H), 7.18 - 7.06 (m, 3H), 6.82 (s, 1H), 6.59 (s, 1H), 5.89 (s, 2H), 4.42 (s, 2H), 3.96 (d, J = 15.1 Hz, 1H), 3.67 (d, J = 15.1 Hz, 1H), 3.57 (dd, J = 11.3, 4.0 Hz, 1H), 3.36 (dd, J = 16.3, 4.2 Hz, 1H), 3.15 (ddd, J = 11.2, 5.6, 2.0 Hz, 1H), 3.06 (ddd, J = 16.8, 11.9, 5.4 Hz, 1H), 2.80 - 2.54 (m, 3H).

[0207] 13 C NMR (100 MHz, Methanol-d4) δ 156.84, 149.35, 146.48, 146.32, 142.79, 137.43, 133.98, 130.17, 129.77, 128.80, 128.12, 127.08, 125.57, 121.81, 119.30, 119.06, 117.82, 116.38, 107.77, 105.05, 100.75, 60.08, 58.05, 50.99, 42.52, 35.23, 28.36.

[0208] Synthesis of compound C15:

[0209]

[0210] The synthetic route of compound C15 is as follows:

[0211]

[0212] According to the preparation method of compound C1, after the reaction of compound 13 (0.32 mmol, 0.10 g) with CDI, DMAP to introduce a carbonyl imidazole group, and then reacted with 4-difluoromethoxybenzylamine (0.81 mmol, 0.14 g), after purification and drying, light yellow solid 30 (compound C15) (0.077 g, 46.9%) was obtained.

[0213] m.p. 113.2-114.5 °C. HRMS (ESI) (M+H) + m / z 510.2222, calcd for C 28 H 30 F2N3O4 + 510.2199.

[0214] 1 H NMR (400 MHz, DMSO-d6) δ 8.46 (s, 1H), 7.38 - 7.33 (d, J = 8.6 Hz, 2H), 7.20 (s, 1H), 7.19 (t, J = 74.4 Hz, 1H), 7.18 - 7.12 (m, 3H), 7.03 (d, J = 8.3 Hz, 1H), 6.89 (s, 1H), 6.69 (s, 1H), 6.61 (t, J = 6.0 Hz, 1H), 4.28 (d, J = 5.8 Hz, 2H), 3.92 (dd, J = 14.4, 4.1 Hz, 1H), 3.74 (d, J = 9.0 Hz, 6H), 3.61 - 3.37 (m, 3H), 3.08 (d, J = 10.4 Hz, 1H), 3.01 - 2.88 (m, 1H), 2.69 - 2.52 (m, 3H).

[0215] 13 C NMR (100 MHz, DMSO-d6) δ 155.74, 150.14, 147.70, 138.55, 138.08, 129.23, 129.11, 119.27, 118.06, 116.88, 115.36, 114.33, 112.18, 109.84, 59.80, 56.21, 55.89, 55.37, 51.23, 42.56, 35.94, 29.49.

[0216] Synthesis of compound C16:

[0217]

[0218] The synthetic route of compound C16 is as follows:

[0219]

[0220] According to the preparation method of compound C1, after the reaction of compound 14 (0.34 mmol, 0.10 g) with CDI, DMAP to introduce carbonyl imidazole group, and then reacted with 4-difluoromethoxy benzylamine (0.85 mmol, 0.15 g), after purification and drying, light yellow solid 31 (compound C16) (0.087 g, 52.0%) was obtained.

[0221] m.p. 176.9-177.4 °C. HRMS (ESI) (M+H) + m / z 494.1905, calcd for C 27 H 26 F2N3O4 + 494.1886.

[0222] 1 H NMR (400 MHz, Methanol-d4) δ 7.41 - 7.34 (m, 2H), 7.22 (d, J = 2.2 Hz, 1H), 7.18 - 7.09 (m, 4H), 6.84 (s, 1H), 6.79 (t, J = 73.2 Hz, 1H), 6.61 (s, 1H), 5.91 (s, 2H), 4.39 (s, 2H), 3.98 (d, J = 15.1 Hz, 1H), 3.69 (d, J = 15.1 Hz, 1H), 3.59 (dd, J = 11.4, 4.0 Hz, 1H), 3.38 (dd, J = 16.3, 4.2 Hz, 1H), 3.17 (ddd, J = 11.2, 5.5, 2.0 Hz, 1H), 3.08 (ddd, J = 16.8, 11.6, 5.3 Hz, 1H), 2.82 - 2.65 (m, 2H), 2.61 (td, J = 11.4, 3.8 Hz, 1H).

[0223] 13C NMR (100 MHz, Methanol-d4) δ 146.49, 146.32, 137.50, 136.62, 133.99, 130.01, 128.78, 128.44, 128.06, 127.11, 118.89, 117.73, 116.29, 107.77, 105.05, 100.75, 60.10, 58.07, 50.99, 42.45, 35.24, 28.37.

[0224] Synthesis of compound C17:

[0225]

[0226] The synthetic route of compound C17 is as follows:

[0227]

[0228] According to the preparation method of compound C1, after the reaction of compound 13 (0.32 mmol, 0.10 g) with CDI, DMAP to introduce a carbonyl imidazole group, and then reacted with 4-methylsulfonylbenzylamine (0.81 mmol, 0.15 g), after purification and drying, light yellow solid 32 (compound C17) (0.084 g, 50.2%) was obtained.

[0229] m.p. 161.7-162.4 °C. HRMS (ESI) (M+H) + m / z 522.2073, calcd for C 28 H 32 N3O5S + 522.2057.

[0230] 1 H NMR (400 MHz, Methanol-d4) δ 7.93 (d, J = 8.3 Hz, 2H), 7.61 (d, J = 8.3 Hz, 2H), 7.26 - 7.20 (m, 1H), 7.20 - 7.10 (m, 2H), 6.91 (s, 1H), 6.73 (s, 1H), 4.51 (s, 2H), 4.06 - 3.95 (m, 1H), 3.85 (d, 6H), 3.71 (d, J = 15.0 Hz, 1H), 3.64 (dd, J = 11.3, 4.1 Hz, 1H), 3.47 (dd, J = 16.3, 4.2 Hz, 1H), 3.25 - 3.14 (m, 1H), 3.17 - 3.05 (m, 4H), 2.85 - 2.69 (m, 2H), 2.64 (td, J = 11.3, 3.8 Hz, 1H).

[0231] 13 C NMR (100 MHz, Methanol-d4) δ 156.83, 147.89, 147.75, 146.70, 139.24, 137.42, 134.01, 129.30, 128.81, 128.23, 127.61, 127.25, 126.33, 117.80, 116.39, 111.56, 109.02, 59.81, 58.09, 55.30, 55.01, 51.11, 43.02, 42.66, 35.07, 27.87.

[0232] Synthesis of compound C18:

[0233]

[0234] The synthetic route of compound C18 is as follows:

[0235]

[0236] According to the preparation method of compound C1, after the reaction of compound 13 (0.32 mmol, 0.10 g) with CDI, DMAP to introduce a carbonyl imidazole group and the reaction with sulfamethoxazole hydrochloride (0.81 mmol, 0.18 g), light yellow solid 33 (compound C18) (0.069 g, 40.9%) was obtained after purification and drying.

[0237] m.p. 184.5-185.1 °C. HRMS (ESI) (M+H) + m / z 523.2026, calcd for C 27 H 31 N4O5S + 523.2010.

[0238] 1 H NMR (400 MHz, Methanol-d4) δ 7.86 (d, J = 8.0 Hz, 2H), 7.49 (d, J = 8.0 Hz, 2H), 7.23 (s, 1H), 7.20 - 7.09 (m, 2H), 6.88 (s, 1H), 6.72 (s, 1H), 4.46 (s, 2H), 4.02 (d, J = 15.2 Hz, 1H), 3.82 (d, J = 9.8 Hz, 6H), 3.79 - 3.66 (m, 2H), 3.47 (dd, J = 16.4, 4.2 Hz, 1H), 3.27 - 3.18 (m, 1H), 3.17 - 3.04 (m, 1H), 2.84 - 2.63 (m, 3H).

[0239] 13C NMR (100 MHz, Methanol-d4) δ 156.84, 147.98, 147.81, 144.62, 142.24, 137.56, 133.51, 128.89, 128.83, 127.90, 127.20, 126.10, 125.98, 117.88, 116.31, 111.54, 108.97, 59.83, 57.88, 55.31, 55.02, 51.02, 42.64, 34.87, 27.66.

[0240] Synthesis of compound C19:

[0241]

[0242] The synthesis route of compound C19 is as follows:

[0243]

[0244] Compound C10 (0.15 mmol, 0.080 g), potassium acetate (0.17 mmol, 0.016 g) were dissolved in 10 mL of ethanol and poured into a solution of iodine (0.15 mmol, 0.038 g) in ethanol, stirred at room temperature for 1 h, then quenched with saturated sodium thiosulfate solution, followed by the addition of sodium hydroxide solution, ethyl acetate was separated, the organic phase was washed with 2M hydrochloric acid solution, saturated sodium chloride solution, respectively, concentrated by rotary evaporation and recrystallized to obtain yellow solid 34 (compound C19) (0.071 g, yield 84.6%).

[0245] m.p. > 280 °C. HRMS (ESI) (M-Cl) + m / z 524.1785, calcd for C 28 H 25 F3N3O4 + 524.1792.

[0246] 1 H NMR (400 MHz, Methanol-d4) δ 9.59 (s, 1H), 8.81 (s, 1H), 8.63 (d, J = 2.2 Hz, 1H), 8.20 (d, J = 9.0 Hz, 1H), 7.98 (dd, J = 9.0, 2.2 Hz, 1H), 7.69 (s, 1H), 7.49 (d, J = 8.7 Hz, 2H) 7.28 (d, J = 8.0 Hz, 2H), 7.08 (s, 1H), 4.90 (t, J = 6.4 Hz, 2H), 4.50 (s, 2H), 4.02 (s, 3H), 3.96 (s, 3H), 3.31 (t, J = 6.4 Hz, 2H).

[0247] 13 C NMR (175 MHz, Methanol-d4) δ 155.91, 152.47, 149.53, 148.18, 147.93, 141.85, 138.78, 135.12, 130.00, 128.61, 128.58, 127.93, 127.37, 120.78, 120.54, 119.95, 119.13, 112.92, 110.84, 108.56, 55.78, 55.59, 55.27, 42.43, 26.34.

[0248] Synthesis of compound C20:

[0249]

[0250] The synthetic route of compound C20 is as follows:

[0251]

[0252] Potassium ferrocyanide (0.90 mmol, 0.29 g) was taken in 5 M 20 mL of sodium hydroxide solution and compound C19 (0.18 mmol, 0.10 g) was added slowly. The reaction was refluxed for 5 h, cooled to room temperature and filtered. The obtained residue was purified by column chromatography (silica gel 100-200 mesh, V(dichloromethane):V(ethanol) = 10:0.1) to get compound 35 (compound C20) as a light yellow solid (0.060 g, yield 61.7%).

[0253] m.p. 248.1-248.7 °C. HRMS (ESI) (M+H) + m / z 540.1741, calcd for C 28 H 25 F3N3O5 + 540.1741. 1 H NMR (400 MHz, DMSO-d6) δ 9.05 (s, 1H), 8.29 (d, J = 2.3 Hz, 1H), 7.78 (dd, J = 8.7, 2.4 Hz, 1H), 7.60 (d, J = 8.6 Hz, 1H), 7.49 - 7.41 (m, 3H), 7.34 (d, J = 8.2 Hz, 2H), 7.21 (s, 1H), 6.95 (s, 1H), 6.84 (t, J = 6.1 Hz, 1H), 4.36 (d, J = 5.9 Hz, 2H), 4.21 (t, J = 6.1 Hz, 2H), 3.88 (s, 3H), 3.82 (s, 3H), 2.91 (t, J = 6.2 Hz, 2H).13 C NMR (175 MHz, DMSO-d6) δ 161.18, 155.63, 150.20, 148.58, 147.57, 140.51, 139.30, 135.46, 131.27, 129.42, 128.50, 127.38, 125.01, 124.12, 122.34, 121.44, 120.58, 114.48, 111.49, 108.66, 101.33, 56.34, 56.08, 42.56, 40.48, 27.72.

[0254] Synthesis of Example 22: The synthetic route for obtaining the precursor of the berberine derivative as shown in general formula II when urea group is attached at C3 position of berberine is shown below:

[0255]

[0256] The reaction reagents and conditions involved in the above synthetic route are as follows: a: borane-tetrahydrofuran complex, THF, 4 h; b: pivaloyl chloride, pyridine, DCM, 5 h; c: stannous chloride, ethanol / water, 5 h; d: bromobenzyl, potassium carbonate, ethanol / water, 0.5 h; e: zinc chloride, acetic anhydride, 24 h; f: ethylene glycol, p-toluenesulfonic acid, toluene, 12 h; g: palladium acetate, Xantphos, potassium phosphate, THF, 3 d; h: ammonium chloride, ethanol / water, 2 d; i: sodium borohydride, methanol, 10 min; j: palladium on carbon, hydrogen, ethanol, 3 h.

[0257] The synthetic route of compound 41 is as follows:

[0258]

[0259] Dissolve m-nitrophenylacetic acid 36 (11.04 mmol, 2.00 g) in 30 mL of tetrahydrofuran, slowly drop borane-tetrahydrofuran complex (1 mol / L, 22.08 mmol, 22.08 mL) at 0°C under argon protection, slowly raise to room temperature after drop completion. After reaction, quench with sodium hydroxide solution, extract with dichloromethane, rotary evaporation, dryness and concentration to obtain m-nitrophenyl ethanol 37. Dissolve 37 in dry dichloromethane, add pyridine (22.08 mmol, 1.74 mL), drop pivaloyl chloride (22.08 mmol, 2.75 mL) at 0°C, add saturated ammonium chloride solution to terminate the reaction after reaction, wash the organic phase with saturated ammonium chloride solution, distilled water, saturated sodium bicarbonate solution, saturated sodium chloride solution respectively, rotary evaporation to obtain light yellow liquid 38 (compound 38) (2.32 g, yield 83.49%).

[0260] Compound 38 (7.96 mmol, 2.00 g) was re-dissolved in 50 mL of ethanol / water 3:1 solution, stannous chloride (31.84 mmol, 7.18 g) was added and the reaction was refluxed for 5 h. After the reaction was completed, 1 M sodium hydroxide solution was added to adjust the pH value to 7, and the organic phase was extracted with excess ethyl acetate and washed with saturated sodium chloride solution, and concentrated to dryness to obtain a light yellow liquid 39 (1.41 g, yield 79.81%). In 50 mL of ethanol / water 3:1 solution, 39 (8.94 mmol, 1.98 g), potassium carbonate (26.83 mmol, 3.71 g) and benzyl bromide (35.77 mmol, 6.12 g) were added and reacted at 80°C for 0.5 h. After the reaction was completed, it was cooled to room temperature, saturated sodium chloride solution, ethyl acetate was added, and the organic phase was washed with saturated sodium bicarbonate solution, sodium chloride solution in turn, dried and concentrated to obtain a yellow oily liquid. Column chromatography (silica gel 200-300 mesh, petroleum ether) gave colorless oily liquid 40 (compound 40) (3.21 g, 89.31%).

[0261] HRMS (ESI) (M+H) + m / z 402.2427, calcd for C 27 H 32 NO2 + 402.2428.

[0262] 1 H NMR (400 MHz, Chloroform-d) δ 7.45 - 7.36 (m, 4H), 7.33 (tt, J = 6.6, 1.4 Hz, 6H), 7.18 (td, J = 7.6, 0.8 Hz, 1H), 6.74 - 6.64 (m, 3H), 4.71 (s, 4H), 4.29 (t, J = 7.1 Hz, 2H), 2.90 (t, J = 7.1 Hz, 2H), 1.24 (s, 9H).

[0263] Compound 40 (2.49 mmol, 1.00 g) was dissolved in 30 mL of acetic anhydride, zinc chloride (12.45 mmol, 1.70 g) was added, and the reaction was carried out at 105°C for 0.5 h under argon atmosphere. After the reaction was completed, distilled water was slowly added dropwise in an ice bath, and extracted with ethyl acetate for 3 times, and the organic phase was combined and washed with saturated sodium bicarbonate solution, and concentrated by rotary evaporation to obtain a brown viscous liquid, which was column chromatographed (silica gel 200-300 mesh, V(petroleum ether):V(ethyl acetate)=15:1) to obtain a light yellow oily liquid compound 41 (0.47 g, yield 42.1%).

[0264] HRMS (ESI) (M+H) + m / z 444.2528, calcd for C29 H 34 NO3 + 444.2533.

[0265] 1 H NMR (400 MHz, Chloroform-d) δ 7.63 (d, J = 8.8 Hz, 1H), 7.29 - 7.18 (m, 6H), 7.15 - 7.10 (m, 4H), 6.55 (d, J = 2.8 Hz, 1H), 6.51 (dd, J = 8.8, 2.8 Hz, 1H), 4.62 (s, 4H), 4.16 (t, J = 6.8 Hz, 2H), 3.15 (t, J = 6.8 Hz, 2H), 2.42 (s, 3H), 1.05 (s, 9H).

[0266] 13 C NMR (100 MHz, Chloroform-d) δ 198.13, 178.50, 151.60, 142.10, 137.25, 133.84, 128.88, 127.32, 126.48, 125.08, 115.37, 109.38, 64.98, 53.64, 38.65, 35.05, 28.62, 27.23.

[0267] The synthetic route of compound 43 is as follows:

[0268]

[0269] Compound 43 (compound 43) (1.14 g, yield 96.3%) was obtained as a white solid after 6-bromo veratraldehyde 42 (4.08 mmol, 1.00 g) was dissolved in 30 mL of toluene, ethylene glycol (8.16 mmol, 0.45 mL), p-toluenesulfonic acid (0.21 mmol, 0.035 g) was added, the reaction was started at 120 °C, after the reaction was completed, it was washed with saturated sodium bicarbonate solution, saturated sodium chloride solution, dried and concentrated.

[0270] HRMS (ESI) (M+H) + m / z 289.0050, calcd for C 11 H 14 BrO4 + 289.0070.

[0271] 1 H NMR (400 MHz, Chloroform-d) δ 7.04 (s, 1H), 6.94 (s, 1H), 5.92 (s, 1H), 4.15 - 3.95 (m, 4H), 3.81 (d, J = 5.7 Hz, 6H).

[0272] The synthetic route of compound 47 is as follows:

[0273]

[0274] Compound 41 (2.25 mmol, 1.00 g), 43 (3.16 mmol, 0.91 g) were dissolved in 10 mL of tetrahydrofuran, and palladium acetate (0.11 mmol, 0.026 g), Xantphos (0.22 mmol, 0.13 g) and potassium phosphate (5.63 mmol, 1.23 g) were added. After the reaction system was replaced with argon, it was reacted at 95 °C for 48 h. After the reaction was completed, it was filtered and concentrated by rotary evaporation to obtain a dry brown oil. Compound 44 (0.68 g, 46.3%) was obtained as a yellow oil by column chromatography (silica gel 200-300 mesh, V(petroleum ether):V(ethyl acetate) = 4:1).

[0275] 1 H NMR (400 MHz, DMSO-d6) δ 7.87 (d, J = 8.7 Hz, 1H), 7.30 (m, 10H), 7.00 (s, 1H), 6.75 (s, 1H), 6.68 - 6.59 (m, 2H), 5.65 (s, 1H), 4.79 (s, 4H), 4.24 (s, 2H), 4.05 (t, J = 6.4 Hz, 2H), 3.93 - 3.77 (m, 4H), 3.71 (d, J = 17.1 Hz, 6H), 3.02 (t, J = 6.8 Hz, 2H), 1.03 (s, 9H).

[0276] Compound 44 (1.04 mmol, 0.68 g) was dissolved in 15 mL of 1 M ammonium chloride solution of ethanol / water (3:1), and reacted at 90 °C for 24 h, and then the temperature was increased to 110 °C and the reaction was continued for 48 h. After the reaction was completed, it was cooled to room temperature and filtered to obtain compound 45 (0.21 g, 38.5%) as a yellow solid.

[0277] 1 H NMR (400 MHz, Methanol-d4) δ 9.09 (s, 1H), 8.29 (s, 1H), 7.77 (d, J = 9.0 Hz, 1H), 7.42 (s, 1H), 7.38 (s, 1H), 7.28 - 7.11 (m, 10H), 6.77 (dd, J = 8.9, 2.7 Hz, 1H), 6.67 (d, J = 2.6 Hz, 1H), 4.73 (s, 4H), 4.60 (t, J = 6.4 Hz, 2H), 3.98 (s, 3H), 3.92 (s, 3H), 3.05 (t, J = 6.3 Hz, 2H).

[0278] Compound 45 (0.40 mmol, 0.21 g) was dissolved in methanol, and sodium borohydride was added to the system at 45 °C. After the reaction was completed, methanol was evaporated, and the residue was dissolved in dichloromethane, washed with water and saturated sodium chloride solution, and concentrated by rotary evaporation to obtain a yellow solid. The yellow solid was purified by column chromatography (silica gel 200-300 mesh, V(dichloromethane):V(ethanol)=10:0.1) to obtain a yellow solid 46 (0.11 g, 57.6%).

[0279] 1 H NMR (400 MHz, Chloroform-d) δ 7.49 - 7.26 (m, 10H), 7.10 (d, J=8.6 Hz, 1H), 6.69-6.65 (m, 2H), 6.59 (s, 1H), 6.54 (d, J=2.7 Hz, 1H), 4.66 (s, 4H), 3.96 (d, J=14.5 Hz, 1H), 3.88 (d, J=2.9 Hz, 6H), 3.67 (d, J=14.6 Hz, 1H), 3.63-3.56 (m, 1H), 3.26 (dd, J=15.9, 3.9 Hz, 1H), 3.20-3.09 (m, 2H), 2.86 (dd, J=15.8, 11.3 Hz, 1H), 2.70-2.59 (m, 2H).

[0280] Compound 46 (0.39 mmol, 0.19 g) was dissolved in ethanol, and 2M 0.5 mL hydrochloric acid solution and palladium-carbon (0.019 g) were added to the system and reacted under a hydrogen atmosphere. After 24 h of reaction, the mixture was filtered and the ethanol was evaporated. The residue was purified by column chromatography (silica gel 200-300 mesh, V(dichloromethane):V(ethanol)=10:0.2) to obtain a yellow solid 47 (0.09 g, 75.6%).

[0281] HRMS (ESI) (M+H) + m / z 311.1751, calcd for C 19 H 23 N2O2 + 311.1754.

[0282] 1H NMR (400 MHz, Chloroform-d) δ 6.99 (d, J = 8.3 Hz, 1H), 6.58 (s, 1H), 6.53 - 6.48 (m, 2H), 6.40 (d, J = 2.4 Hz, 1H), 3.89 (d, J = 14.5 Hz, 1H), 3.78 (d, J = 2.7 Hz, 6H), 3.68 - 3.49 (m, 2H), 3.18 (dd, J = 16.1, 4.1 Hz, 1H), 3.12 - 3.03 (m, 2H), 2.76 (t, J = 13.7 Hz, 1H), 2.65 - 2.52 (m, 2H).

[0283] Synthesis of compound C21:

[0284]

[0285] The synthetic route of compound C21 is as follows:

[0286]

[0287] According to the preparation method of compound C1, after the reaction of compound 47 (0.32 mmol, 0.10 g) with CDI, DMAP to introduce a carbonyl imidazole group, and then reacted with 4-aminomethyltetrahydro pyran (0.81 mmol, 0.092 g), after purification and drying, yellow solid 48 (compound C21) (0.081 g, 55.9%) was obtained.

[0288] m.p. 131.7 - 132.4 °C. HRMS (ESI) (M+H) + m / z 452.2541, calcd for C 26 H 34 N3O4 + 452.2544.

[0289] 1 H NMR (400 MHz, Methanol-d4) δ 7.16 - 7.08 (m, 2H), 7.05 (d, J = 2.0 Hz, 1H), 6.67 (s, 1H), 6.60 (s, 1H), 3.90 - 3.81 (m, 3H), 3.70 (d, J = 2.8 Hz, 6H), 3.59 - 3.48 (m, 2H), 3.36 - 3.25 (m, 3H), 3.12 - 2.97 (m, 4H), 2.72 - 2.61 (m, 2H), 2.54 (td, J = 10.9, 10.3, 3.7 Hz, 1H), 1.66 (dtt, J = 11.4, 8.0, 3.6 Hz, 1H), 1.56 (d, J = 13.1 Hz, 2H), 1.27 - 1.13 (m, 2H).

[0290] 13 C NMR (175 MHz, Methanol-d4) δ 156.99, 148.00, 147.70, 137.94, 134.22, 131.10, 126.06, 125.64, 125.60, 118.43, 117.19, 111.69, 109.25, 67.36, 59.70, 57.52, 55.10, 55.07, 51.01, 45.05, 35.66, 35.13, 30.33, 28.44.

[0291] Synthesis of compound C22:

[0292]

[0293] The synthetic route of compound C22 is as follows:

[0294]

[0295] According to the preparation method of compound C1, after reacting with CDI, DMAP to introduce carbonyl imidazole group, compound 47 (0.32 mmol, 0.10 g) was reacted with 4-aminomethyl propionyl piperidine (0.81 mmol, 0.14 g), and after drying by purification, light yellow solid 49 (compound C22) (0.076 g, 46.8%) was obtained.

[0296] m.p. 126.7-127.1 °C. HRMS (ESI) (M+H) + m / z 507.2969, calcd for C 29 H 39 N4O4 + 507.2966.

[0297] 1H NMR (400 MHz, Methanol-d4) δ 7.30 - 7.21 (m, 2H), 7.20 (d, J = 2.1 Hz, 1H), 6.81 (s, 1H), 6.74 (s, 1H), 4.56 (d, J = 13.7 Hz, 1H), 4.07 - 3.95 (m, 2H), 3.83 (d, J = 3.0 Hz, 6H), 3.78 - 3.70 (m, 2H), 3.47 (dd, J = 16.5, 4.3 Hz, 1H), 3.31 - 3.22 (m, 1H), 3.22 - 3.03 (m, 4H), 2.87 - 2.68 (m, 3H), 2.70 - 2.59 (m, 1H), 2.43 (q, J = 7.5 Hz, 2H), 1.88 - 1.75 (m, 3H), 1.21 (dd, J = 12.5, 4.1 Hz, 1H), 1.13 (t, J = 7.5 Hz, 3H).

[0298] 13 C NMR (175 MHz, Methanol-d4) δ 173.25, 156.95, 148.10, 147.77, 138.07, 134.02, 130.71, 125.85, 125.66, 125.14, 118.42, 117.25, 111.65, 109.21, 59.74, 57.35, 55.11, 55.06, 50.94, 45.40, 44.54, 41.54, 36.80, 34.96, 30.08, 29.27, 28.26, 25.99, 8.65.

[0299] Synthesis of compound C23:

[0300]

[0301] The synthetic route of compound C23 is as follows:

[0302]

[0303] According to the preparation method of compound C1, after the reaction of compound 47 (0.32 mmol, 0.10 g) with CDI, DMAP to introduce a carbonyl imidazole group, and then with 4-trifluoromethoxybenzylamine (0.81 mmol, 0.15 g), light yellow solid 50 (compound C23) (0.10 g, 60.3%) was obtained after purification and drying.

[0304] m.p. 136.7-137.2 °C. HRMS (ESI) (M+H) + m / z 528.2103, calcd for C 28 H29 F3N3O4 + 528.2105.

[0305] 1 H NMR (400 MHz, Methanol-d4) δ 7.32 (d, J = 8.3 Hz, 2H), 7.16 - 7.08 (m, 5H), 6.68 (s, 1H), 6.61 (s, 1H), 4.30 (s, 2H), 3.93 (d, J = 15.3 Hz, 1H), 3.70 (d, J = 3.0 Hz, 6H), 3.69 - 3.60 (m, 2H), 3.35 (dd, J = 16.5, 4.1 Hz, 1H), 3.20 - 3.12 (m, 1H), 3.10-2.99 (m, 1H), 2.75 - 2.60 (m, 3H).

[0306] 13 C NMR (175 MHz, Methanol-d4) δ 156.75, 148.22, 148.10, 147.85, 139.15, 138.12, 133.81, 130.38, 128.49, 125.70, 125.60, 124.61, 120.73, 120.54, 119.82, 118.55, 117.44, 111.64, 109.19, 59.74, 57.12, 55.11, 55.08, 50.84, 42.37, 34.73, 28.03.

[0307] Synthesis of compound C24:

[0308]

[0309] The synthetic route of compound C24 is as follows:

[0310]

[0311] According to the preparation method of compound C1, after the reaction of compound 47 (0.32 mmol, 0.10 g) with CDI, DMAP to introduce a carbonyl imidazole group, and the reaction with 4-difluoromethoxybenzylamine (0.81 mmol, 0.14 g), after purification and drying, light yellow solid 51 (compound C24) (0.091 g, 55.2%) was obtained.

[0312] m.p. 188.2-188.9 °C. HRMS (ESI) (M+H) + m / z 510.2198, calcd for C 28 H 30 F2N3O4 +510.2199.

[0313] 1 H NMR (400 MHz, DMSO-d6) δ 8.51 (s, 1H), 7.35 (d, J = 8.4 Hz, 2H), 7.20 (t, J = 74.2 Hz, 1H), 7.23 - 7.11 (m, 5H), 6.73 (s, 1H), 6.70 - 6.61 (m, 2H), 4.28 (d, J = 5.9 Hz, 2H), 3.87 (d, J = 14.8 Hz, 1H), 3.72 (s, 6H), 3.56 - 3.39 (m, 2H), 3.29 (dd, J = 16.0, 3.9 Hz, 1H), 3.10 - 3.02 (m, 1H), 2.97 (ddd, J = 16.5, 11.4, 5.4 Hz, 1H), 2.67 - 2.52 (m, 2H), 2.50 - 2.40 (m, 1H).

[0314] 13 C NMR (175 MHz, DMSO-d6) δ 155.74, 150.13, 147.63, 147.49, 138.70, 138.09, 134.99, 131.44, 129.20, 126.85, 126.61, 126.18, 119.26, 117.79, 116.89, 116.46, 112.34, 109.91, 59.56, 58.00, 55.98, 55.93, 51.24, 42.54, 36.27, 29.63.

[0315] Synthesis of compound C25:

[0316]

[0317] The synthetic route of compound C25 is as follows:

[0318]

[0319] According to the preparation method of compound C19, compound C23 (0.15 mmol, 0.080 g), potassium acetate in ethanol solution and iodine in ethanol solution were mixed and reacted, and after purification and drying, yellow solid 52 (compound C25) (0.068 g, yield 81.1%) was obtained.

[0320] m.p. > 240 °C. HRMS (ESI) (M-Cl) + m / z 524.1794, calcd for C 28 H 25 F3N3O4 + 524.1792.

[0321] 1 H NMR (400 MHz, Methanol-d4) δ 9.23 (s, 1H), 8.49 (s, 1H), 7.94 (d, J = 8.7 Hz, 1H), 7.55 - 7.47 (m, 3H), 7.44 (dd, J = 8.6, 2.2 Hz, 1H), 7.35 (d, 2H), 7.15 (d, 2H), 4.70 (t, J = 6.4 Hz, 2H), 4.34 (s, 2H), 4.03 (s, 3H), 3.96 (s, 3H), 3.17 (t, J = 6.4 Hz, 2H).

[0322] 13 C NMR (175 MHz, Methanol-d4) δ 158.57, 156.08, 153.20, 148.13, 144.70, 143.29, 139.20, 138.92, 137.59, 135.85, 128.55, 126.52, 123.25, 120.75, 120.54, 120.36, 117.76, 116.89, 105.87, 104.91, 56.06, 55.63, 54.82, 42.36, 27.21.

[0323] The technical effects and advantages of the present application are verified by the application of the berberine derivative of the present application in an anti-inflammatory drug for inhibiting inflammation caused by soluble epoxide hydrolase.

[0324] Specifically, the verification method of the anti-inflammatory drug in the embodiments of the present application is to incubate the berberine derivative with a sample containing soluble epoxide hydrolase and the endogenous hydrolysis substrate 14, 15-EET of the hydrolysis enzyme, detect the content of the hydrolysis product 14, 15-DHET, and use the relative amount of generation to reflect the inhibition of the compound on soluble epoxide hydrolase. The specific steps are as follows:

[0325] Brain tissue samples from adult (25-30 g body weight) male C57BL / 6J mice were collected and added with 1:10 (mg / μL) pre-cooled PBS (phosphate buffer saline) (pH = 7.4) and homogenized for 30 seconds (4°C, 4500 rpm) using a homogenizer (Bertin Precellys 24-Dual, Bertin Technologies), then centrifuged at 9000 g for 15 minutes at 4°C using a small low-temperature centrifuge, and the supernatant was diluted 20 times. In an EP (eppendorf) tube, 160 μL of PBS and 20 μL of the diluted tissue liquid and 2 μL of the compound with a concentration gradient of 10000, 5000, 1000, 500, 100, 50, 10, 1, 0.1, 0.01 μg / mL (final concentration 100, 50, 10, 5, 1, 0.5, 0.1, 0.01, 0.001, 0.0001 μg / mL) were added, and incubated at room temperature for 15 minutes. The negative control was PBS without the tissue diluent but with the solvent (180 μL of PBS plus 2 μL of DMSO), and the positive control was the sample diluent with the same concentration plus the same volume of solvent (160 μL of PBS plus 20 μL of the diluted tissue liquid plus 2 μL of DMSO). After the incubation at room temperature, all samples were placed on ice, and the enzyme reaction substrate (10 μL of 14, 15-EET (10 μg / mL)) was quickly added and mixed, and then incubated in a shaking incubator at 37°C. After the incubation, the samples were placed on ice, and 10 μL of 800 nM t-TUCB (No. 6757, Tocris Bioscience, UK, Bristol) was quickly added to terminate the reaction, and the final reaction system volume was 200 μL. In order to detect the enzyme activity, the concentration of the reaction product 14, 15-DHET was detected using high performance liquid chromatography-mass spectrometry. The pretreatment was as follows: 1. An equal volume (200 μL) of a mixture of methanol and acetonitrile (50 / 50, v / v) was added to the sample, and the mixture contained 0.4% acetic acid and internal standards 11, 12-EET-d11 and 11, 12-DHET-d11 (20 ng / mL); 2. The mixture was mixed using a homogenizer at 4500 rpm for 10 minutes, and was placed at -20°C for 2 hours to precipitate the protein; 3. After centrifugation of the sample for 10 minutes (14000 rpm), the supernatant was collected for UPLC-MS / MS analysis. The relative enzyme activity of sEH was calculated by comparing the amount of 14, 15-DHET produced by the tested compound at a certain concentration with the amount of 14, 15-DHET produced without the inhibitor, and the inhibition rate was 100% minus the relative enzyme activity.

[0326] wherein the inhibition rate % (5 μg / mL) is the inhibition rate of the compound on sEH at 5 μg / mL.

[0327] The control compounds are TPPU, EC5026 and BBR.

[0328] The specific verification results of the berberine derivatives prepared in the embodiments of the present application are shown in Table 1.

[0329] Table 1. Inhibition rate and IC50 of the berberine derivatives prepared in the embodiments of the present application on sEH

[0330]

[0331]

[0332]

[0333]

[0334] The specific verification results of the control compounds TPPU, EC5026 and BBR on soluble epoxide hydrolase are shown in Table 2.

[0335] Table 2. Inhibition rate and IC50 of the control compounds on sEH under the same conditions

[0336]

[0337] As can be seen from Table 1 and Table 2, compared with the control compound berberine BBR, the berberine derivatives prepared in the present application have excellent soluble epoxide hydrolase (sEH) inhibitory activity, and the IC50 values of some of the berberine derivatives are better than 43.77 nM of the positive control compound TPPU and 19.11 nM of the positive control compound EC5026, and the berberine has excellent sEH inhibitory activity by successful structural modification. The unique tertiary amine structure of the berberine skeleton greatly increases its drugability, and has a great advantage compared with the skeletons of other sEH inhibitors. Overall, the berberine skeleton has the best activity by connecting the benzyl group containing different substituents through a urea group, and the activity is equivalent by connecting an alkyl chain and a heterocycle; the different oxidation states of the C ring of the berberine skeleton result in great differences in spatial conformation, and the tetrahydro structure is better than the quaternary ammonium salt structure which is better than the lactam structure; the activity of the urea group connected to the C-10 position of the berberine skeleton is better than that of the C-3 position. Therefore, the present application obtains a novel soluble epoxide hydrolase inhibitor compound by introducing the berberine skeleton described in general formula I and II, and the IC50 values of most of the compounds on soluble epoxide hydrolase are less than 20 nM, and the lowest IC50 value is about 0.67 nM.

[0338] To sum up, the prepared berberine derivative for inhibiting soluble epoxide hydrolase has a main skeleton of berberine with a urea group, different substituents are connected to different positions of the berberine skeleton through the urea group, and the oxidation state of the C ring is changed, so that a series of berberine derivatives with high activity on soluble epoxide hydrolase are obtained, and the IC50 value of the berberine derivative for inhibiting soluble epoxide hydrolase reaches the nanomolar level, indicating that the berberine derivative has high inhibition effect on soluble epoxide hydrolase, so that the inhibition effect on inflammation can be achieved by inhibiting soluble epoxide hydrolase, and the berberine derivative has good application prospect.

Claims

1. A berberine derivative, characterized by, The berberine derivative has a general structure of formula I or formula II: Formula I Formula II In the formula I or II, R 1 and R 2 are both methoxy, or R 1 and R 2 form a ring to form methylenedioxy; R 3 any one of isobutyl, tetrahydrofurylmethyl, tetrahydropyranylmethyl, cyclohexylmethyl, N-propionylpiperidinyl, N-propionylpiperidinylmethyl, trifluoromethoxyphenyl, difluoromethoxybenzyl, trifluoromethoxybenzyl, methylsulfonylbenzyl, aminosulfonylbenzyl; The C ring is selected from any one of the following structures: or or .

2. The berberine derivative according to claim 1, characterized by R 1 , R 2 form a methylenedioxy group.

3. The berberine derivative according to claim 1, characterized by, said R 3 is any one group selected from isobutyl, tetrahydrofurylmethylidene, tetrahydropyranylmethylidene, N-propionylpiperidinyl, N-propionylpiperidinylmethylidene, trifluoromethoxyphenyl, difluoromethoxybenzyl, trifluoromethoxybenzyl, methylsulfonylbenzyl, aminosulfonylbenzyl.

4. The berberine derivative according to claim 3, characterized by said R 3 is selected from any one of trifluoromethoxybenzyl, difluoromethoxybenzyl.

5. The berberine derivative according to claim 1, wherein The C ring is selected from any one of the following structures: or .

6. The berberine derivative according to claim 5, characterized in that, The C ring is selected from .

7. The berberine derivative according to claim 1, wherein R 1 and R 2 form a methylenedioxy group or R 1 , R 2 are each methoxy, R 3 is selected from any one of tetrahydrofuranylmethylene, N-propionylpiperidinylmethylene, trifluoromethoxybenzyl, difluoromethoxybenzyl, methanesulfonylbenzyl or aminosulfonylbenzyl, and ring C is .

8. The berberine derivative according to claim 1, wherein The berberine derivative has a general structure of formula I or formula II: 。 9. A pharmaceutical composition of a berberine derivative, characterized by, The pharmaceutical composition comprises the berberine derivative according to any one of claims 1-8.

10. Use of a berberine derivative as defined in any one of claims 1 to 8, or a pharmaceutical composition of a berberine derivative as defined in claim 9, for the manufacture of a medicament for the treatment of inflammation, wherein, The anti-inflammatory drug is used for inhibiting inflammation caused by soluble epoxide hydrolase.

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