Tetrahydroberberine derivative and application thereof

Abstract

The invention discloses a tetrahydroberberine derivative and application of the tetrahydroberberine derivative. The tetrahydroberberine derivative is a # imgabs0 # compound as shown in a general formula (I). According to the present invention, tetrahydroberberine is adopted as a substrate, organic chemical structure modification and derivatization are performed to obtain the 12-substituted tetrahydroberberine derivative, and compared with tetrahydroberberine, the 12-substituted aryl or heteroaromatic tetrahydroberberine derivative provides an inhibition effect on Erastatin-induced HK-2 cell ferroptosis, and can effectively inhibit ferroptosis-induced acute kidney injury. The compound can be applied to preparation of related drugs for preventing, relieving and / or treating ferroptosis related diseases, and a new direction is provided for preventing, relieving and / or treating ferroptosis related diseases.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and particularly relates to a class of tetrahydroberberine derivatives and their applications. Background Art

[0002] Ferroptosis is a regulated form of cell death characterized by the accumulation of intracellular lipid peroxides to lethal levels, and this process depends on the participation of iron. Different from other types of cell death (such as apoptosis or necrosis), ferroptosis is mainly triggered by the imbalance of intracellular redox homeostasis, especially the peroxidation reaction of polyunsaturated fatty acids (PUFAs) in cell membranes. Iron acts as a catalyst, exacerbating the generation of reactive oxygen species (ROS), thus further promoting the process of lipid peroxidation. Ferroptosis has received extensive attention due to its involvement in various diseases, especially in neurodegenerative diseases, cancer, and ischemic organ injuries, especially in pathological conditions such as acute kidney injury (AKI). Given that ferroptosis has a unique molecular mechanism, it has become a potential therapeutic target for regulating cell death, especially in diseases where the traditional apoptosis mechanism is insufficient to play a role. Therefore, it is of great significance to develop inhibitors related to ferroptosis. Summary of the Invention

[0003] Aiming at the deficiencies of the prior art, the purpose of the present invention is to provide a class of tetrahydroberberine derivatives, their preparation methods and applications. Specifically, taking tetrahydroberberine as a substrate, through organic chemical structure modification and derivatization, a class of 12-substituted tetrahydroberberine derivatives is obtained, and their application in the preparation of related drugs for preventing, alleviating and / or treating ferroptosis-related diseases provides a new option for preventing, alleviating and / or treating ferroptosis-related diseases.

[0004] To solve the problems of the prior art, the technical solutions adopted by the present invention are as follows:

[0005] A class of tetrahydroberberine derivatives, the general formula is as shown in (I) as shown, wherein

[0006] When X = N or O, Y = (CH 2 )n, n = 0 - 6, R is one of an aromatic ring, a heteroaromatic ring, a monosubstituted or polysubstituted aromatic ring and a heteroaromatic ring, specifically as shown;

[0007] When X = Y = 0, R is one of an aromatic ring and its monosubstituted or polysubstituted aromatic ring, a heteroaromatic ring and its monosubstituted or polysubstituted heteroaromatic ring, specifically as shown;

[0008] When X = N, O, R is one of an aromatic ring, a heteroaromatic ring, a monosubstituted or polysubstituted aromatic ring and a heteroaromatic ring, specifically as as shown

[0009] As an improvement, when X = Y = 0, R is p-chlorophenyl, p-fluorophenyl or p-methylphenyl.

[0010] As an improvement, when X = N, O, 2 Y = (CH )n, n = 0 - 3 or a four-membered, five-membered or aromatic ring containing a heteroatom, R is p-chlorophenyl, p-fluorophenyl, p-methylphenyl, phenyl, p-hydroxyphenyl, p-fluoropyridine, 1,2,4-trifluorophenyl, 4-hydroxymethylfuryl, 4-fluoro-2-chlorophenyl, 2,4-dichlorophenyl, 2,4,6-trifluorophenyl, 2,6-difluoro-4-hydroxyphenyl, 4-acetamidophenyl or 4-cyanophenyl.

[0011] As an improvement, when X = N, O, R is p-fluorophenyl.

[0012] As an improvement, the salt is selected from hydrobromide, hydroiodide, hydrofluoride, hydrochloride, sulfate, nitrate, phosphate, citrate, acetate, lactate.

[0013] Further improved, the salt is hydrobromide, hydrochloride or sulfate.

[0014] The above-mentioned type of tetrahydroberberine derivatives specifically include: 12-(N-(4-hydroxybenzyl))aminotetrahydroberberine; 12-(N-(2,4-dihydroxybenzyl))aminotetrahydroberberine; 12-(N-(4-acetamidobenzyl))aminotetrahydroberberine; 12-(N-(3-hydroxymethylfuran))aminotetrahydroberberine; 12-(N-(2,4-dichlorobenzyl))aminotetrahydroberberine; 12-(N-(2,4,6-trifluorobenzyl))aminotetrahydroberberine; 12-(N-(4-hydroxy-2,6-difluorobenzyl))aminotetrahydroberberine; 12-(N-(4-fluorobenzyl))aminotetrahydroberberine; 12-(N-(2,4-difluorobenzyl))aminotetrahydroberberine; 12-(N-(2,4,5-trifluorobenzyl))aminotetrahydroberberine; 12-(N-(2-dichloro-4-fluoro-benzyl))aminotetrahydroberberine; 12-(N-(2-fluoropyridine-5-methylene-))aminotetrahydroberberine; 12-(N-(4-cyanobenzyl))aminotetrahydroberberine; 12-(N-benzyl)aminotetrahydroberberine; 12-(N-(4-chlorobenzyl))aminotetrahydroberberine; 12-(N-(4-methylbenzyl))aminotetrahydroberberine; 12-(N-methyl,N-(4-fluorobenzyl))aminotetrahydroberberine; 12-(N-(2-(4-fluorophenyl)ethyl))aminotetrahydroberberine; 12-(N-(2-(4-fluorophenyl)propyl))aminotetrahydroberberine; 12-(N-(4-fluorophenyl))aminotetrahydroberberine; N-(4'-fluoro-1,1'-biphenyl-3-yl)aminotetrahydroberberine; 12-(N-(1-(p-fluorophenyl)pyrrolidin-3-yl))aminotetrahydroberberine; N-(6-(4-fluorophenyl)pyridin-2-yl)aminotetrahydroberberine; N-(4'-methyl-1,1'-biphenyl-4-yl)aminotetrahydroberberine; N-(4'-chloro-1,1'-biphenyl-4-yl)aminotetrahydroberberine; 12-(N-(1-(m-fluorophenyl)pyrrolidin-3-yl))aminotetrahydroberberine; 12-(N-(1-(o-fluorophenyl)pyrrolidin-3-yl))aminotetrahydroberberine; 12-(N-(1-(p-chlorophenyl)pyrrolidin-3-yl))aminotetrahydroberberine; 12-(N-(1-(p-methylphenyl)pyrrolidin-3-yl))aminotetrahydroberberine; N-(6-fluoronaphthalen-2-yl)aminotetrahydroberberine; N-(4'-methyl-1,1'-biphenyl-4-yl)aminotetrahydroberberine; 12-p-chlorophenyltetrahydroberberine; 12-p-methylphenyltetrahydroberberine; 12-p-fluorophenyltetrahydroberberine; 12-(p-fluorobenzamide)tetrahydroberberine.

[0015] The structural formulas corresponding to the above compounds are shown in the following table:

[0016] Structure 1:

[0017]

[0018]

[0019]

[0020] The specific tetrahydroberberine derivatives of the above-mentioned type are as follows:

[0021] Structure 2:

[0022]

[0023] The specific tetrahydroberberine derivatives of the above-mentioned type are as follows:

[0024] Structure 3:

[0025]

[0026] An inhibitor, the active ingredient of which is the above-mentioned tetrahydroberberine derivative.

[0027] A pharmaceutical composition comprising the above-mentioned tetrahydroberberine derivative.

[0028] The above-mentioned tetrahydroberberine derivative, the above-mentioned inhibitor, and the above-mentioned pharmaceutical composition are used in the preparation of a medicament for preventing, alleviating, and / or treating ferroptosis-related diseases.

[0029] Beneficial effects:

[0030] Compared with the prior art, for the tetrahydroberberine derivatives of the first type of the present invention and their preparation methods and applications, the tetrahydroberberine derivatives with a 12-substituted aryl or heteroaryl ring have a good inhibitory effect on ferroptosis-induced acute kidney injury. Through experiments, it is found that compared with tetrahydroberberine, the tetrahydroberberine derivatives with a 12-substituted aryl or heteroaryl ring have an inhibitory effect on ferroptosis induced by Erastin in HK-2 cells, and can more effectively inhibit ferroptosis-induced acute kidney injury, and have good application prospects. Specifically, compared with the protective effect of THB on ferroptosis induced by Erastin in HK-2 cells, the activity of most THB derivatives has increased by nearly 100 times. Description of the drawings

[0031] Figure 1 Shows the effects of compounds 20, 21, 30, and 31 on the ROS value in HK-2 cells after treatment with Erastin at different concentrations;

[0032] Figure 2 Shows the effects of compounds 20, 21, 30, and 31 on effectively reducing the MDA value in HK-2 cells after treatment with Erastin at different concentrations;

[0033] Figure 3 Kidney sections of mice after treatment with compounds 20, 21, 30, and 31 for folic acid-induced acute kidney injury in mice;

[0034] Figure 4 Effect of compounds 20, 21, 30, and 31 on the mRNA expression of kidney injury molecule-1 (KIM-1) in mice with acute kidney injury;

[0035] Figure 5 Effect of compounds 20, 21, 30, and 31 on the mRNA expression of neutrophil gelatinase-associated lipocalin (NGAL) in mice with acute kidney injury, which can effectively reduce it;

[0036] Figure 6 Effect of compounds 20, 21, 30, and 31 on the content of serum creatinine (SCR) in mice with acute kidney injury;

[0037] Figure 7 Effect of compounds 20, 21, 30, and 31 on the content of blood urea nitrogen (BUN) in mice with acute kidney injury. Detailed implementation manners

[0038] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of this application. Obviously, the described embodiments are some, but not all, of the embodiments of this application. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of this application without creative efforts shall fall within the protection scope of this application.

[0039] A class of tetrahydroberberine derivatives, with the general formula as shown in (I) as follows, where

[0040] When X = Y = 0, R is one of an aromatic ring, a mono- or poly-substituted aromatic ring, a heteroaromatic ring, and a mono- or poly-substituted heteroaromatic ring, specifically as shown in as follows;

[0041] When X = N or O, Y = (CH 2 )n, n = 0 - 6, and R is one of an aromatic ring, a heteroaromatic ring, a mono- or poly-substituted aromatic ring, and a mono- or poly-substituted heteroaromatic ring, specifically as shown in as follows;

[0042] When X = N, O, R is one of an aromatic ring, a heteroaromatic ring, a mono- or poly-substituted aromatic ring, and a mono- or poly-substituted heteroaromatic ring, specifically as shown in as follows.

[0043] As an improvement, when X = Y = 0, R is p-chlorophenyl, p-fluorophenyl, or p-methylphenyl.

[0044] As an improvement, when X = N or O, and Y = (CH 2 )n where n = 0 - 3, or a four-membered, five-membered, or aromatic ring containing a heteroatom, R is p-chlorophenyl, p-fluorophenyl, p-methylphenyl, phenyl, p-hydroxyphenyl, p-fluoropyridine, 1,2,4-trifluorophenyl, 4-hydroxymethylfuryl, 4-fluoro-2-chlorophenyl, 2,4-dichlorophenyl, 2,4,6-trifluorophenyl, 2,6-difluoro-4-hydroxyphenyl, 4-acetamidophenyl, or 4-cyanophenyl.

[0045] As an improvement, when X = N or O, R is p-fluorophenyl.

[0046] As an improvement, the salt is selected from hydrobromide, hydroiodide, hydrofluoride, hydrochloride, sulfate, nitrate, phosphate, citrate, acetate, and lactate.

[0047] Further improvement is that the salt is hydrobromide, hydrochloride, or sulfate.

[0048] The above-mentioned class of tetrahydroberberine derivatives specifically include: 12-(N-(4-hydroxybenzyl))aminotetrahydroberberine; 12-(N-(2,4-dihydroxybenzyl))aminotetrahydroberberine; 12-(N-(4-acetamidobenzyl))aminotetrahydroberberine; 12-(N-(3-hydroxymethylfuran))aminotetrahydroberberine; 12-(N-(2,4-dichlorobenzyl))aminotetrahydroberberine; 12-(N-(2,4,6-trifluorobenzyl))aminotetrahydroberberine; 12-(N-(4-hydroxy-2,6-difluorobenzyl))aminotetrahydroberberine; 12-(N-(4-fluorobenzyl))aminotetrahydroberberine; 12-(N-(2,4-difluorobenzyl))aminotetrahydroberberine; 12-(N-(2,4,5-trifluorobenzyl))aminotetrahydroberberine; 12-(N-(2-dichloro-4-fluoro-benzyl))aminotetrahydroberberine; 12-(N-(2-fluoropyridine-5-methylene-))aminotetrahydroberberine; 12-(N-(4-cyanobenzyl))aminotetrahydroberberine; 12-(N-benzyl)aminotetrahydroberberine; 12-(N-(4-chlorobenzyl))aminotetrahydroberberine; 12-(N-(4-methylbenzyl))aminotetrahydroberberine; 12-(N-methyl,N-(4-fluorobenzyl))aminotetrahydroberberine; 12-(N-(2-(4-fluorophenyl)ethyl))aminotetrahydroberberine; 12-(N-(2-(4-fluorophenyl)propyl))aminotetrahydroberberine; 12-(N-(4-fluorophenyl))aminotetrahydroberberine; N-(4'-fluoro-1,1'-biphenyl-3-yl)aminotetrahydroberberine; 12-(N-(1-(p-fluorophenyl)pyrrolidin-3-yl))aminotetrahydroberberine; N-(6-(4-fluorophenyl)pyridin-2-yl)aminotetrahydroberberine; N-(4'-methyl-1,1'-biphenyl-4-yl)aminotetrahydroberberine; N-(4'-chloro-1,1'-biphenyl-4-yl)aminotetrahydroberberine; 12-(N-(1-(m-fluorophenyl)pyrrolidin-3-yl))aminotetrahydroberberine; 12-(N-(1-(o-fluorophenyl)pyrrolidin-3-yl))aminotetrahydroberberine; 12-(N-(1-(p-chlorophenyl)pyrrolidin-3-yl))aminotetrahydroberberine; 12-(N-(1-(p-methylphenyl)pyrrolidin-3-yl))aminotetrahydroberberine; N-(6-fluoronaphthalen-2-yl)aminotetrahydroberberine; N-(4'-methyl-1,1'-biphenyl-4-yl)aminotetrahydroberberine; 12-p-chlorophenyltetrahydroberberine; 12-p-methylphenyltetrahydroberberine; 12-p-fluorophenyltetrahydroberberine; 12-(p-fluorobenzamide)tetrahydroberberine.

[0049] The above-mentioned class of tetrahydroberberine derivatives specifically include:

[0050]

[0051]

[0052] An inhibitor, the active ingredient of which is a tetrahydroberberine derivative.

[0053] The above inhibitor is used in the preparation of a medicament for preventing, alleviating and / or treating ferroptosis-related diseases.

[0054] Example 1

[0055] The reaction formula of the compound of Structural Formula 1 is as follows:

[0056]

[0057] Preparation of Intermediate 2:

[0058] Weigh berberine hydrochloride (30.00 g, 80.47 mmol) into a reaction flask, add 200 mL of ethanol, and add NaBH 4 (30.44 g, 800.47 mmol) portionwise at 0 °C, and react at 0 °C for 4 h. Quench with water, filter, and dry to obtain tetrahydroberberine, 24.50 g of yellowish-green solid, with a yield of 89.8%. 1H NMR (300 MHz, Chloroform-d) δ 6.77–6.70 (m, 2H), 6.67 (d, J = 8.3 Hz, 1H), 6.60 (s, 1H), 5.92 (s, 2H), 4.26 (d, J = 15.6 Hz, 1H), 3.87 (s, 3H), 3.58 (s, 2H), 3.30–3.07 (m, 3H), 2.95–2.53 (m, 3H).

[0059] Preparation of Intermediate 3:

[0060] Place tetrahydroberberine (24.50 g, 72.27 mmol) in a reaction flask, add 200 mL of glacial acetic acid, and add NaNO 2 (24.93 g, 361.35 mmol) portionwise at 0 °C, then dropwise add 50 mL of concentrated nitric acid, and react at 25 °C for 6 h. Quench with concentrated ammonia water under an ice bath, filter, dry the filter cake, and perform column chromatography, DCM:MeOH = 100:1, to obtain 12-nitrotetrahydroberberine, a red or yellow solid (9.60 g, 25.00 mmol), with a yield of 34.6%. 11H NMR (300 MHz, Chloroform-d) δ 7.63 (s, 1H), 6.75 (s, 1H), 6.60 (s, 1H), 5.94 (s, 2H), 4.26 (d, J = 16.1 Hz, 1H), 3.96 (s, 3H), 3.93 (s, 3H), 3.58–3.44 (m, 3H), 3.28–3.04 (m, 4H), 2.74–2.60 (m, 2H).

[0061] Preparation of Intermediate 4:

[0062] Dissolve 12-nitroberberine (9.60 g, 25.00 mmol) in a mixed solvent of 100 mL MeOH:THF = 1:1, add 6H 2 O·NiCl 2 , then add NaBH 4 (9.45 g, 250.00 mol) portionwise, react at ice bath for 0.5 h, rotary evaporate to dryness, dissolve with DCM, filter with Buchner funnel, rotary evaporate the filtrate to dryness, column chromatography, PE:EA = 1:1, to obtain 12-aminoberberine, yellow solid (5.2 g, 14.6 mmol), yield 58.4%. 1H NMR (300 MHz, DMSO) δ 6.97 (s, 1H), 6.67 (s, 1H), 6.24 (s, 1H), 5.96 (dd, J = 5.1, 1.0 Hz, 2H), 4.71 (s, 2H), 3.97 (d, J = 15.6 Hz, 1H), 3.69 (s, 3H), 3.59 (s, 3H), 3.27 (d, J = 15.7 Hz, 2H), 3.07 (dt, J = 15.7, 4.9 Hz, 2H), 2.96–2.84 (m, 1H), 2.61 (d, J = 15.7 Hz, 1H), 2.46–2.33 (m, 1H), 2.10 (dd, J = 15.9, 11.1 Hz, 1H).

[0063] Synthesis of Compound 1 in Example 1-1

[0064] Weigh intermediate 4 (80.00 mg, 0.23 mmol), p-hydroxybenzaldehyde (28.06 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 g, 0.046 mmol) into a reaction flask, add 3 mL of methanol, stir at 80 °C for 6 h, quench with water, extract three times with DCM, combine the organic phases, dry over anhydrous sodium sulfate, column chromatography, PE:EA = 10:3, to obtain 38 mg of pale yellowish-white solid, yield 36.5%. 11H NMR (300 MHz, DMSO) δ 9.22 (s, 1H), 7.27–7.12 (m, 2H), 7.02 (s, 1H), 6.70 (d, J = 8.3 Hz, 3H), 6.04 (s, 1H), 5.96 (dd, J = 8.3, 1.0 Hz, 2H), 5.48 (t, J = 6.0 Hz, 1H), 4.21 (d, J = 5.8 Hz, 2H), 3.96 (d, J = 15.6 Hz, 1H), 3.61 (s, 3H), 3.57 (s, 3H), 3.37 (s, 1H), 3.27 (d, J = 15.5 Hz, 1H), 3.17 (d, J = 5.2 Hz, 1H), 3.09 (dd, J = 11.8, 4.4 Hz, 1H), 2.92 (t, J = 14.0 Hz, 1H), 2.61 (d, J = 15.8 Hz, 1H), 2.47–2.35 (m, 1H), 2.16 (dd, J = 15.9, 11.0 Hz, 1H). MS [M+1]: 461.2

[0065] Synthesis of Compound 2 in Example 1-2

[0066] Weigh intermediate 4 (80.00 mg, 0.23 mmol), 2,4-dihydroxybenzaldehyde (31.00 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 mg, 0.046 mmol) into a reaction flask, add 3 mL of methanol, stir at 80 °C for 6 h, quench with water, extract three times with DCM, combine the organic phases, dry over anhydrous sodium sulfate, and perform column chromatography with PE:EA = 10:3 to obtain 45 mg of a pale yellowish-white solid with a yield of 41.8%. 1 1H NMR (400 MHz, Chloroform-d) δ 6.71 (s, 1H), 6.65 (d, J = 8.6 Hz, 1H), 6.56 (d, J = 9.2 Hz, 2H), 6.37 (d, J = 2.9 Hz, 1H), 6.35 (s, 1H), 5.89 (dd, J = 8.3, 1.4 Hz, 2H), 4.22 (t, J = 15.0 Hz, 2H), 4.04 (d, J = 13.5 Hz, 1H), 3.76 (s, 3H), 3.75 (s, 3H), 3.63–3.54 (m, 2H), 3.26–3.12 (m, 2H), 2.97 (dd, J = 15.5, 4.3 Hz, 1H), 2.71–2.60 (m, 2H), 2.45 (dd, J = 15.4, 10.9 Hz, 1H). MS [M+1]: 477.2

[0067] Synthesis of Compound 3 in Example 1-3

[0068] Weigh intermediate 4 (80.00 mg, 0.23 mmol), 4-acetamidobenzaldehyde (29.00 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 mg, 0.046 mmol) into a reaction flask, add 3 mL of methanol, stir at 80 °C for 6 h, quench with water, extract three times with DCM, combine the organic phases, dry over anhydrous sodium sulfate, and perform column chromatography with PE:EA = 10:3 to obtain 42 mg of a pale yellowish-white solid with a yield of 37.1%. 1 1H NMR (400 MHz, Chloroform-d) δ 7.47 (d, J = 8.1 Hz, 2H), 7.31 (t, J = 9.5 Hz, 2H), 6.73 (s, 1H), 6.58 (s, 1H), 6.13 (s, 1H), 5.93–5.87 (m, 2H), 4.30 (s, 2H), 4.19 (d, J = 15.7 Hz, 1H), 3.75 (s, 3H), 3.75 (s, 3H), 3.59–3.49 (m, 2H), 3.22–3.04 (m, 2H), 2.91 (dd, J = 15.3, 4.3 Hz, 1H), 2.70–2.56 (m, 2H), 2.38 (dd, J = 15.1, 10.9 Hz, 1H), 2.16 (s, 3H). MS [M+1]: 502.2

[0069] Synthesis of Compound 4 in Examples 1-4

[0070] Weigh intermediate 4 (80.00 mg, 0.23 mmol), 4-(hydroxymethyl)furan-2-carbaldehyde (23.00 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 mg, 0.046 mmol) into a reaction flask, add 3 mL of methanol, stir at 80 °C for 6 h, quench with water, extract three times with DCM, combine the organic phases, dry over anhydrous sodium sulfate, and perform column chromatography with PE:EA = 10:3 to obtain 54 mg of a brownish-red solid with a yield of 51%. 11H NMR (300 MHz, DMSO) δ 7.00 (s, 1H), 6.68 (s, 1H), 6.28–6.22 (m, 2H), 6.19 (d, J = 3.1 Hz, 1H), 5.96 (dd, J = 6.0, 1.0 Hz, 2H), 5.43 (t, J = 5.9 Hz, 1H), 5.15 (t, J = 5.7 Hz, 1H), 4.34 (d, J = 5.6 Hz, 2H), 4.28 (d, J = 5.7 Hz, 2H), 3.98 (d, J = 15.6 Hz, 1H), 3.72 (s, 3H), 3.59 (s, 3H), 3.28 (d, J = 15.6 Hz, 2H), 3.17–3.04 (m, 2H), 2.99–2.84 (m, 1H), 2.61 (d, J = 15.7 Hz, 1H), 2.47–2.35 (m, 1H), 2.12 (dd, J = 15.9, 11.0 Hz, 1H). MS [M+1]: 465.2

[0071] Synthesis of Compound 5 in Examples 1 - 5

[0072] Weigh intermediate 4 (80.00 mg, 0.23 mmol), 2,4 - dichlorobenzaldehyde (39.00 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 mg, 0.046 mmol) into a reaction flask, add 3 mL of methanol, stir at 80 °C for 6 h, quench with water, extract three times with DCM, combine the organic phases, dry over anhydrous sodium sulfate, and perform column chromatography with PE:EA = 10:3 to obtain 45 mg of a pale yellow - white solid with a yield of 38.9%. 1H NMR (300 MHz, Chloroform - d) δ 7.37 (d, J = 2.5 Hz, 1H), 7.32 (d, J = 8.5 Hz, 1H), 7.19 (dd, J = 8.5, 2.6 Hz, 1H), 6.77 (s, 1H), 6.61 (s, 1H), 6.00 (s, 1H), 5.93 (s, 2H), 4.42 (s, 2H), 4.21 (d, J = 15.6 Hz, 1H), 3.93 (s, 1H), 3.76 (s, 3H), 3.74 (s, 3H), 3.58 (s, 2H), 3.17 (s, 2H), 2.99 (d, J = 14.8 Hz, 1H), 2.68 (d, J = 16.1 Hz, 2H), 2.46 (s, 1H). MS [M+1]: 513.1

[0073] Synthesis of Compound 6 in Examples 1 - 6

[0074] Weigh intermediate 4 (80.00 mg, 0.23 mmol), 2,4,6-trifluorobenzaldehyde (36.00 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 mg, 0.046 mmol) into a reaction flask, add 3 mL of methanol, stir at 80 °C for 6 h, quench with water, extract three times with DCM, combine the organic phases, dry over anhydrous sodium sulfate, and perform column chromatography with PE:EA = 10:3 to obtain 36 mg of a pale yellowish-white solid with a yield of 32.0%. 1H NMR (300 MHz, Chloroform-d) δ 6.71 (d, J = 14.3 Hz, 1H), 6.65 (d, J = 8.2 Hz, 1H), 6.60 (s, 1H), 6.35 (s, 1H), 5.94 (s, 2H), 4.41 (s, 2H), 4.20 (s, 1H), 3.86 (s, 3H), 3.75 (s, 3H), 3.54 (s, 2H), 3.16 (s, 2H), 2.89 (d, J = 15.0 Hz, 1H), 2.66 (s, 2H), 2.36 (s, 1H). MS [M+1]: 499.2

[0075] Synthesis of Compound 7 in Examples 1 - 7

[0076] Weigh intermediate 4 (80.00 mg, 0.23 mmol), 2,6-difluoro-4-hydroxybenzaldehyde (36.00 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2(6.27 mg, 0.046 mmol) was placed in a reaction flask, 3 mL of methanol was added, and the mixture was stirred at 80 °C for 6 h. The reaction was quenched by adding water, and the mixture was extracted three times with DCM. The organic phases were combined, dried over anhydrous sodium sulfate, and purified by column chromatography (PE:EA = 10:3) to obtain 47 mg of a pale yellowish-white solid with a yield of 41.9%. 1H NMR (300 MHz, DMSO) δ 6.97 (s, 1H), 6.67 (s, 1H), 6.42 (d, J = 9.7 Hz, 2H), 6.24 (s, 1H), 5.95 (dd, J = 11.5, 1.0 Hz, 2H), 5.77 (s, 1H), 5.17–5.09 (m, 1H), 4.21 (d, J = 5.5 Hz, 2H), 3.96 (d, J = 15.6 Hz, 1H), 3.74 (s, 3H), 3.59 (s, 3H), 3.39 (s, 1H), 3.24 (s, 1H), 3.12–2.99 (m, 2H), 2.96–2.82 (m, 1H), 2.60 (d, J = 15.5 Hz, 1H), 2.46–2.33 (m, 1H), 2.06 (dd, J = 15.9, 11.1 Hz, 1H). MS [M+1]: 497.2

[0077] Synthesis of Compound 8 in Examples 1 - 8

[0078] Weighed intermediate 4 (80.00 mg, 0.23 mmol), p-fluorobenzaldehyde (28.00 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 mg, 0.046 mmol) was placed in a reaction flask, 3 mL of methanol was added, and the mixture was stirred at 80 °C for 6 h. The reaction was quenched by adding water, and the mixture was extracted three times with DCM. The organic phases were combined, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain 70 mg of a brown solid with a yield of 67%. 11H NMR (400 MHz, Chloroform-d) δ 7.39–7.33 (m, 2H), 7.09–7.02 (m, 2H), 6.76 (s, 1H), 6.62 (s, 1H), 6.15 (s, 1H), 5.93 (q, J = 1.5 Hz, 2H), 4.35 (s, 2H), 4.25 (d, J = 15.7 Hz, 1H), 3.79 (s, 3H), 3.78 (s, 3H), 3.66–3.54 (m, 2H), 3.29–3.21 (m, 1H), 3.14 (ddd, J = 15.0, 10.4, 4.6 Hz, 1H), 2.95 (ddd, J = 15.3, 4.4, 1.2 Hz, 1H), 2.74–2.61 (m, 2H), 2.44 (ddd, J = 15.3, 11.0, 1.5 Hz, 1H). MS [M+1]: 463.2

[0079] Synthesis of Compound 9 in Examples 1 - 9

[0080] Weigh intermediate 4 (80.00 mg, 0.23 mmol), 2,4-difluorobenzaldehyde (32.00 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 mg, 0.046 mmol) into a reaction flask, add 3 mL of methanol, stir at 80 °C for 6 h, quench with water, extract three times with DCM, combine the organic phases, dry over anhydrous sodium sulfate, and perform column chromatography with PE:EA = 10:3 to obtain 34 mg of a red solid with a yield of 31.3%. 1 1H NMR (400 MHz, Chloroform-d) δ 7.39–7.33 (m, 2H), 7.09–7.02 (m, 2H), 6.76 (s, 1H), 6.62 (s, 1H), 6.15 (s, 1H), 5.93 (q, J = 1.5 Hz, 2H), 4.35 (s, 2H), 4.25 (d, J = 15.7 Hz, 1H), 3.79 (s, 3H), 3.78 (s, 3H), 3.66–3.54 (m, 2H), 3.29–3.21 (m, 1H), 3.14 (ddd, J = 15.0, 10.4, 4.6 Hz, 1H), 2.95 (ddd, J = 15.3, 4.4, 1.2 Hz, 1H), 2.74–2.61 (m, 2H), 2.44 (ddd, J = 15.3, 11.0, 1.5 Hz, 1H). MS [M+1]: 481.2

[0081] Synthesis of Compound 10 in Examples 1 - 10

[0082] Weigh intermediate 4 (80.00 mg, 0.23 mmol), 2,4,5-trifluorobenzaldehyde (36.00 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 mg, 0.046 mmol) into a reaction flask, add 3 mL of methanol, stir at 80 °C for 6 h, quench with water, extract three times with DCM, combine the organic phases, dry over anhydrous sodium sulfate, perform column chromatography, PE:EA = 10:3, to obtain 38 mg of a white solid with a yield of 33.8%. 1 1H NMR (300 MHz, DMSO) δ 7.18 (t, J = 8.7 Hz, 2H), 6.94 (s, 1H), 6.66 (s, 1H), 6.22 (s, 1H), 5.94 (dd, J = 12.0, 1.0 Hz, 2H), 5.27 (t, J = 5.7 Hz, 1H), 4.30 (d, J = 5.6 Hz, 2H), 3.95 (d, J = 15.6 Hz, 1H), 3.73 (s, 3H), 3.58 (s, 3H), 3.24 (s, 2H), 3.05 (s, 2H), 2.86 (d, J = 13.1 Hz, 1H), 2.61 (s, 1H), 2.39 (t, J = 9.7 Hz, 1H), 2.14–1.97 (m, 1H). MS[M+1]: 499.2 Synthesis of compound 11 in Examples 1 - 11

[0083] Weigh intermediate 4 (80.00 mg, 0.23 mmol), 2-chloro-4-fluorobenzaldehyde (36.00 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 mg, 0.046 mmol) into a reaction flask, add 3 mL of methanol, stir at 80 °C for 6 h, quench with water, extract three times with DCM, combine the organic phases, dry over anhydrous sodium sulfate, perform column chromatography, PE:EA = 10:3, to obtain 50 mg of a light greenish-white solid with a yield of 44.6%. 11H NMR (300 MHz, Chloroform-d) δ 7.34 (dd, J = 8.6, 6.1 Hz, 1H), 7.15 (dd, J = 8.4, 2.6 Hz, 1H), 6.94 (td, J = 8.3, 2.6 Hz, 1H), 6.76 (s, 1H), 6.61 (s, 1H), 6.05 (s, 1H), 5.92 (s, 2H), 4.42 (s, 2H), 4.21 (d, J = 15.6 Hz, 1H), 3.76 (s, 3H), 3.75 (s, 3H), 3.58 (s, 2H), 3.19 (s, 2H), 2.97 (dd, J = 15.5, 4.2 Hz, 1H), 2.68 (d, J = 15.6 Hz, 2H), 2.45 (s, 1H).

[0084] MS [M+1]: 497.2

[0085] Synthesis of Compound 12 in Example 1 - 12

[0086] Weigh intermediate 4 (80.00 mg, 0.23 mmol), 5-fluoropyridine-2-carbaldehyde (28.06 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 mg, 0.046 mmol) into a reaction flask, add 3 mL of methanol, stir at 80 °C for 6 h, quench with water, extract three times with DCM, combine the organic phases, dry over anhydrous sodium sulfate, and perform column chromatography with PE:EA = 10:3 to obtain 45 mg of a red solid with a yield of 43.0%. 1 1H NMR (300 MHz, DMSO) δ 8.29 (d, J = 2.3 Hz, 1H), 8.00 (td, J = 8.3, 2.5 Hz, 1H), 7.14 (dd, J = 8.4, 2.8 Hz, 1H), 7.01 (s, 1H), 6.69 (s, 1H), 6.06 (s, 1H), 5.97 (d, J = 9.3 Hz, 2H), 5.66 (t, J = 6.2 Hz, 1H), 4.38 (d, J = 6.0 Hz, 2H), 3.97 (d, J = 15.6 Hz, 1H), 3.63 (s, 3H), 3.57 (s, 3H), 3.28 (d, J = 15.5 Hz, 1H), 3.12 (t, J = 17.7 Hz, 2H), 2.89 (d, J = 12.0 Hz, 1H), 2.62 (d, J = 15.8 Hz, 1H), 2.47–2.34 (m, 1H), 2.18 (dd, J = 15.9, 11.1 Hz, 1H). MS [M+1]: 464.2

[0087] Synthesis of Compound 13 in Example 1 - 13

[0088] Weigh intermediate 4 (80.00 mg, 0.23 mmol), p-cyanobenzaldehyde (30.00 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 mg, 0.046 mmol) into a reaction flask, add 3 mL of methanol, stir at 80 °C for 6 h, quench with water, extract three times with DCM, combine the organic phases, dry over anhydrous sodium sulfate, and perform column chromatography with PE:EA = 10:3 to obtain 36 mg of a pale red-white solid with a yield of 33.9%. 1 1H NMR (300 MHz, DMSO) δ 7.82–7.76 (m, 2H), 7.60 (d, J = 8.0 Hz, 2H), 7.02 (s, 1H), 6.69 (s, 1H), 5.98 (d, J = 1.0 Hz, 1H), 5.97–5.91 (m, 2H), 5.79 (t, J = 5.9 Hz, 1H), 4.44 (d, J = 6.0 Hz, 2H), 3.97 (d, J = 15.6 Hz, 1H), 3.56 (d, J = 1.5 Hz, 6H), 3.26 (s, 1H), 3.23–3.06 (m, 2H), 2.92 (t, J = 14.1 Hz, 1H), 2.62 (d, J = 15.8 Hz, 1H), 2.43 (t, J = 11.5 Hz, 1H), 2.21 (dd, J = 15.9, 11.1 Hz, 1H). MS [M+1]: 470.2

[0089] Synthesis of Compound 14 in Examples 1 - 14

[0090] Weigh intermediate 4 (80.00 mg, 0.23 mmol), benzaldehyde (24.00 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 mg, 0.046 mmol) into a reaction flask, add 3 mL of methanol, stir at 80 °C for 6 h, quench with water, extract three times with DCM, combine the organic phases, dry over anhydrous sodium sulfate, and perform column chromatography with PE:EA = 10:3 to obtain 46 mg of a yellow-green solid with a yield of 45.8%. 11H NMR (300 MHz, DMSO) δ 7.41 (d, J = 7.1 Hz, 2H), 7.35–7.28 (m, 2H), 7.25–7.17 (m, 1H), 7.02 (s, 1H), 6.69 (s, 1H), 6.01 (s, 1H), 5.97 (dd, J = 7.8, 1.0 Hz, 2H), 5.64 (t, J = 6.1 Hz, 1H), 4.34 (d, J = 5.9 Hz, 2H), 3.97 (d, J = 15.6 Hz, 1H), 3.58 (s, 3H), 3.56 (s, 3H), 3.28 (d, J = 15.7 Hz, 2H), 3.17 (dt, J = 12.6, 4.2 Hz, 1H), 3.09 (dd, J = 11.6, 4.3 Hz, 1H), 2.92 (t, J = 13.9 Hz, 1H), 2.62 (d, J = 15.7 Hz, 1H), 2.48–2.36 (m, 1H), 2.19 (dd, J = 15.9, 11.0 Hz, 1H). MS [M+1]: 445.2

[0091] Synthesis of Compound 15 in Examples 1 - 15

[0092] Weigh intermediate 4 (80.00 mg, 0.23 mmol), p - chlorobenzaldehyde (32.00 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 mg, 0.046 mmol) into a reaction flask, add 3 mL of methanol, stir at 80 °C for 6 h, quench with water, extract three times with DCM, combine the organic phases, dry over anhydrous sodium sulfate, and perform column chromatography with PE:EA = 10:3 to obtain 46 mg of a white solid with a yield of 41.3%. 1 1H NMR (300 MHz, DMSO) δ 7.43 (d, J = 8.4 Hz, 2H), 7.37 (d, J = 8.5 Hz, 2H), 7.01 (s, 1H), 6.69 (s, 1H), 5.99–5.96 (m, 2H), 5.95 (s, 1H), 5.69 (t, J = 6.1 Hz, 1H), 4.33 (d, J = 6.0 Hz, 2H), 3.96 (d, J = 15.5 Hz, 1H), 3.58 (s, 3H), 3.57 (s, 3H), 3.25 (s, 2H), 3.21–3.11 (m, 1H), 3.08 (s, 1H), 2.89 (d, J = 13.0 Hz, 1H), 2.62 (d, J = 15.7 Hz, 1H), 2.40 (d, J = 11.3 Hz, 1H), 2.19 (dd, J = 15.8, 11.0 Hz, 1H). MS [M+1]: 479.2

[0093] Synthesis of Compound 16 in Example 1-16

[0094] Weigh intermediate 4 (80.00 mg, 0.23 mmol), p-tolualdehyde (27.00 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 mg, 0.046 mmol) into a reaction flask, add 3 mL of methanol, stir at 80 °C for 6 h, quench with water, extract three times with DCM, combine the organic phases, dry over anhydrous sodium sulfate, and perform column chromatography with PE:EA = 10:3 to obtain 52 mg of a yellow-green solid with a yield of 50.2%. 1 1H NMR (300 MHz, DMSO) δ 7.29 (d, J = 7.9 Hz, 2H), 7.12 (d, J = 7.8 Hz, 2H), 7.02 (s, 1H), 6.69 (s, 1H), 6.01 (s, 1H), 5.97 (dd, J = 8.0, 1.0 Hz, 2H), 5.58 (t, J = 6.0 Hz, 1H), 4.28 (d, J = 5.9 Hz, 2H), 3.96 (d, J = 15.6 Hz, 1H), 3.59 (s, 3H), 3.56 (s, 3H), 3.28 (d, J = 15.5 Hz, 2H), 3.16 (dd, J = 16.1, 4.5 Hz, 1H), 3.11–3.04 (m, 1H), 2.92 (t, J = 13.8 Hz, 1H), 2.62 (d, J = 15.8 Hz, 1H), 2.48–2.35 (m, 1H), 2.26 (s, 3H), 2.17 (dd, J = 15.9, 11.0 Hz, 1H). MS [M+1]: 459.2

[0095] Synthesis of Compound 17 in Example 1-17

[0096] Weigh intermediate 4 (80.00 mg, 0.23 mmol), p-fluorobenzaldehyde (28.06 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2(6.27 mg, 0.046 mmol) was placed in a reaction flask, 3 mL of methanol was added, and the mixture was stirred at 80 °C for 6 h. The reaction was quenched by adding water, and the mixture was extracted with DCM three times. The organic phases were combined, dried over anhydrous sodium sulfate, and purified by column chromatography (PE:EA = 10:3) to obtain 70 mg of a pale yellowish-green solid. 50 mg of the solid obtained above was weighed, and ZnCl2 and NaBH3CN were weighed and added to a single-necked flask. Then, 21 formaldehyde solution (37%) was added dropwise, and the mixture was stirred overnight at room temperature. The mixture was extracted with DCM three times. The organic phases were combined, dried over anhydrous sodium sulfate, and purified by column chromatography (PE:EA = 5:1) to obtain 42 mg of a white solid with a yield of 39.0%. 1 H NMR (400 MHz, Chloroform-d) δ 7.39–7.33 (m, 2H), 7.09–7.02 (m, 2H), 6.76 (s, 1H), 6.62 (s, 1H), 6.15 (s, 1H), 5.93 (q, J = 1.5 Hz, 2H), 4.35 (s, 2H), 4.25 (d, J = 15.7 Hz, 1H), 3.79 (s, 3H), 3.78 (s, 3H), 3.66–

[0097] 3.54 (m, 2H), 3.29–3.21 (m, 1H), 3.14 (ddd, J = 15.0, 10.4, 4.6 Hz, 1H), 2.95 (ddd, J = 15.3, 4.4, 1.2 Hz, 1H), 2.74–2.61 (m, 2H), 2.44 (ddd, J = 15.3, 11.0, 1.5 Hz, 1H). MS [M+1]: 477.2

[0098] Synthesis of Compound 18 in Example 1-18

[0099] Intermediate 4 (80.00 mg, 0.23 mmol), 4-fluorophenylacetaldehyde (31.00 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 mg, 0.046 mmol) were placed in a reaction flask, 3 mL of methanol was added, and the mixture was stirred at 80 °C for 6 h. The reaction was quenched by adding water, and the mixture was extracted with DCM three times. The organic phases were combined, dried over anhydrous sodium sulfate, and purified by column chromatography (PE:EA = 10:3) to obtain 36 mg of a yellowish-green solid with a yield of 33.4%. 11H NMR (300 MHz, DMSO) δ 7.37–7.27 (m, 2H), 7.17–7.08 (m, 2H), 6.96 (s, 1H), 6.68 (s, 1H), 6.16 (s, 1H), 5.97 (dd, J = 7.6, 1.0 Hz, 2H), 4.84 (t, J = 5.7 Hz, 1H), 3.99 (d, J = 15.6 Hz, 1H), 3.77 (s, 3H), 3.61 (s, 3H), 3.31 (d, J = 5.7 Hz, 2H), 3.27 (d, J = 2.6 Hz, 2H), 3.14–2.98 (m, 2H), 2.97–2.83 (m, 3H), 2.61 (d, J = 15.8 Hz, 1H), 2.47–2.35 (m, 1H), 2.08 (dd, J = 15.8, 11.0 Hz, 1H). MS [M+1]: 477.2

[0100] Synthesis of Compound 19 in Examples 1 - 19

[0101] Weigh intermediate 4 (80.00 mg, 0.23 mmol), 4-fluorobenzaldehyde (34.00 mg, 0.23 mmol), NaBH 3 CN (43.36 mg, 0.69 mmol), ZnCl 2 (6.27 mg, 0.046 mmol) into a reaction flask, add 3 mL of methanol, stir at 80 °C for 6 h, quench with water, extract three times with DCM, combine the organic phases, dry over anhydrous sodium sulfate, and perform column chromatography with PE:EA = 10:3 to obtain 54 mg of a pale yellowish-white solid with a yield of 48.7%. 1 1H NMR (300 MHz, DMSO) δ 7.34–7.23 (m, 2H), 7.16–7.06 (m, 2H), 7.00 (s, 1H), 6.67 (s, 1H), 6.03 (s, 1H), 5.96 (dd, J = 9.0, 1.0 Hz, 2H), 4.78 (t, J = 5.6 Hz, 1H), 3.98 (d, J = 15.5 Hz, 1H), 3.71 (s, 3H), 3.60 (s, 3H), 3.28 (d, J = 15.5 Hz, 2H), 3.08 (q, J = 8.7 Hz, 4H), 2.92–2.83 (m, 1H), 2.69 (t, J = 7.5 Hz, 2H), 2.61 (d, J = 15.6 Hz, 1H), 2.46–2.34 (m, 1H), 2.10 (dd, J = 15.9, 11.1 Hz, 1H), 1.88 (p, J = 7.4 Hz, 2H). MS [M+1]: 491.2

[0102] Example 2

[0103] The synthetic route of Structural Formula 1 is as follows:

[0104]

[0105] Synthesis of Compound 20 in Example 2-1

[0106] Weigh intermediate 4 (80.00 mg, 0.23 mmol), p-bromofluorobenzene (39.00 mg, 0.23 mmol), BINAP (14.00 mg, 0.023 mmol), Pd 2 (dba) 3 (21.00 mg, 0.023 mmol), and cesium carbonate (65.00 mg, 0.69 mmol) into a two-necked flask. Add 3 mL of anhydrous 1,4-dioxane and stir at 110 °C for 12 h. After monitoring the reaction to completion by TLC, quench with water, extract three times with DCM, and perform column chromatography with PE:EA = 10:3 to obtain 20 mg of a pale yellow solid with a yield of 20%. 1 H NMR (300 MHz, DMSO) δ 7.25 (s, 1H), 7.20–6.98 (m, 2H), 6.90 (d, J = 5.0 Hz, 1H), 6.87 (d, J = 6.4 Hz, 2H), 6.69 (d, J = 6.2 Hz, 2H), 5.94 (d, J = 11.2 Hz, 2H), 4.06 (d, J = 15.8 Hz, 1H), 3.71 (s, 3H), 3.70 (s, 3H), 3.26–3.07 (m, 3H), 2.89 (d, J = 13.0 Hz, 1H), 2.61 (d, J = 15.8 Hz, 2H), 2.43 (d, J = 11.6 Hz, 1H), 2.36–2.23 (m, 1H). MS [M+1]: 449.2

[0107] Synthesis of Compound 21 in Example 2-2

[0108] Weigh p-fluorobenzeneboronic acid (200.00 mg, 1.43 mmol), p-dibromobenzene (300.00 mg, 1.43 mmol), Pd(PPh 3 ) 4 (165 mg, 0.143 mmol), and Cs 2 CO 3 (931 mg, 1.43 mmol) into a two-necked flask. N 2Evacuate and exhaust for 5 minutes, then add 8 mL of 1,4-dioxane. Stir at 110 °C for 6 h. After the reaction is completed, quench with water, extract three times with DCM, and perform column chromatography on a PE column to obtain 178 mg of a colorless liquid with a yield of 52.5%. Weigh the above colorless liquid (57.00 mg, 0.23 mmol), 12-aminotetrahydroberberine (80.00 mg, 0.23 mmol), BINAP (14.00 mg, 0.023 mmol), Pd 2 (dba) 3 (21.00 mg, 0.023 mmol), t-BuONa (43.00 mg, 0.45 mmol) into a two-necked flask. Evacuate and exhaust for 5 minutes, add 3 mL of anhydrous 1,4-dioxane, stir overnight at 110 °C to obtain 42 mg of a red-brown solid with a yield of 35%. 2 Evacuate and exhaust for 5 minutes, add 3 mL of anhydrous 1,4-dioxane, stir overnight at 110 °C to obtain 42 mg of a red-brown solid with a yield of 35%. 1 1H NMR (300 MHz, DMSO) δ 7.65–7.56 (m, 2H), 7.44 (s, 1H), 7.27 (td, J = 8.3, 5.8 Hz, 3H), 7.12 (t, J = 2.0 Hz, 1H), 7.00–6.96 (m, 1H), 6.91–6.83 (m, 3H), 6.68 (s, 1H), 5.93 (dd, J = 12.8, 1.0 Hz, 2H), 4.08 (d, J = 15.9 Hz, 1H), 3.72 (d, J = 2.0 Hz, 6H), 3.44 (s, 2H), 3.25 (dd, J = 16.2, 3.9 Hz, 1H), 3.16–3.08 (m, 1H), 2.98–2.85 (m, 1H), 2.62 (d, J = 15.7 Hz, 1H), 2.47–2.25 (m, 2H). MS [M+1]: 525.2

[0109] Synthesis of Compound 22 in Example 2-3

[0110] Weigh 12-aminotetrahydroberberine, 1-tert-butoxycarbonyl-3-pyrrolidone, ZnCl 2 , NaBH 3 CN and add them to 3 mL of methanol. Stir at 80 °C and monitor by TLC until the reaction is complete. Quench with water, extract three times with DCM, evaporate to dryness, add 6 mL of 1,4-dioxane hydrochloride, stir at room temperature for 6 h, adjust to alkaline with 5 M NaOH, extract three times with DCM, evaporate to dryness, and perform column chromatography, DCM:MeOH = 50:1, DCM:MeOH:Et 3 N = 10:1:0.2 to obtain 197 mg of a gray solid. Weigh the above gray solid (180.00 mg, 0.43 mmol), p-bromofluorobenzene (74.00 mg, 0.43 mmol), BINAP (39.00 mg, 0.043 mmol), Pd2 (dba) 3 (26.00 mg, 0.043 mmol), t-BuONa (80.00 mg, 0.85 mmol) were placed in a double-necked bottle, stirred at 80°C, monitored by TLC until the reaction was complete, quenched by adding water, extracted three times with DCM, and chromatographed by reverse phase column, MeOH:H2O=35:65-90:10, to obtain 48 mg of compound 21 as a light reddish white solid, with a yield of 22%, and 39 mg of compound 22 as a light reddish white solid, with a yield of 18%. MS [M+1]: 518.2

[0111] Example 2-4 Synthesis of Compound 23

[0112] Weigh p-fluorophenylboric acid (200.00 mg, 1.43 mmol), 2,5-dibromopyridine (339.00 mg, 1.43 mmol), Pd(PPh 3 ) 4 (165.00 mg, 0.143 mmol), Cs 2 CO 3 (932 mg, 2.86 mmol) was placed in a two-necked bottle, N 2 After evacuation for 5 minutes, 8 mL of 1,4-dioxane was added and stirred at 110°C for 6 h. After the reaction was completed, water was added to quench the reaction, and DCM was extracted three times. PE column chromatography was performed to obtain 252 mg of a light yellow solid with a yield of 69.7%. The above light yellow solid (57.00 mg, 0.23 mmol), 12-aminotetrahydroberberine (80.00 mg, 0.23 mmol), BINAP (14.00 mg, 0.023 mmol), Pd 2 (dba) 3 (21.00 mg, 0.023 mmol), t-BuONa (43.00 mg, 0.45 mmol) were placed in a two-necked bottle, N 2 The mixture was evacuated for 5 minutes, 3 mL of anhydrous 1,4-dioxane was added, and the mixture was stirred at 110° C. overnight to obtain compound 23 as a yellow solid (31 mg). The yield was 25.7%. 1H NMR (300MHz, DMSO) δ8.14–8.04(m,3H),7.65–7.56(m,2H),7.28(td,J=7.9,2.6H z,3H),6.92(s,1H),6.78(d,J=8.2Hz,1H),6.68(s,1H),5.94(d,J=12.4Hz,2H), 4.09(d,J=15.8Hz,1H),3.78(s,3H),3.74(s,3H),3.13(d,J=10.1Hz,1H),2.92( t,J=12.3Hz,1H),2.62(d,J=15.8Hz,1H),2.41(d,J=11.0Hz,2H).MS[M+1]:526.2

[0113] Example 2-5 Synthesis of Compound 24

[0114] Weigh p-methylphenylboronic acid (200.00 mg, 1.43 mmol), p-dibromobenzene (344.00 mg, 1.43 mmol), Pd(PPh 3 ) 4 (165 mg, 0.143 mmol), Cs 2 CO 3 (931 mg, 1.43 mmol) was placed in a two-necked bottle, N 2 After evacuation for 5 minutes, 8 mL of 1,4-dioxane was added and stirred at 110°C for 6 h. After the reaction was completed, water was added to quench the reaction, and DCM was extracted three times. PE column chromatography was performed to obtain 216 mg of brown-gray solid with a yield of 61.4%. The above brown-gray solid (57.00 mg, 0.23 mmol), 12-aminotetrahydroberberine (80.00 mg, 0.23 mmol), BINAP (14.00 mg, 0.023 mmol), Pd 2 (dba) 3 (21.00 mg, 0.023 mmol), t-BuONa (43.00 mg, 0.45 mmol) were placed in a two-necked bottle, N 2 The mixture was evacuated for 5 minutes, 3 mL of anhydrous 1,4-dioxane was added, and the mixture was stirred at 110° C. overnight to obtain compound 24 as a yellow solid (42 mg). The yield was 35.7%. 11H NMR (300 MHz, DMSO) δ 7.51 (s, 1H), 7.32–7.26 (m, 2H), 6.84–6.73 (m, 4H), 6.67 (s, 1H), 5.93 (dd, J = 12.8, 1.0 Hz, 2H), 4.07 (d, J = 15.9 Hz, 1H), 3.73 (s, 3H), 3.72 (s, 3H), 3.43 (s, 2H), 3.24–3.06 (m, 2H), 2.98–2.83 (m, 1H), 2.61 (d, J = 15.8 Hz, 1H), 2.46–2.39 (m, 1H), 2.28 (dd, J = 16.0, 11.4 Hz, 1H). MS [M+1]: 521.2

[0115] Synthesis of Compound 25 in Example 2 - 6

[0116] Weigh p-chlorophenylboronic acid (200.00 mg, 1.28 mmol), p-dibromobenzene (300.00 mg, 1.28 mmol), Pd(PPh 3 ) 4 (144.00 mg, 0.128 mmol), Cs 2 CO 3 (835 mg, 1.28 mmol) into a two-necked flask. Evacuate with N 2 for 5 minutes, then add 8 mL of 1,4-dioxane. Stir at 110 °C for 6 h. After the reaction is completed, quench with water and extract three times with DCM. Purify by PE column chromatography to obtain 235 mg of a white solid with a yield of 69.0%. Weigh the above white solid (60.00 mg, 0.23 mmol), 12-aminotetrahydroberberine (80.00 mg, 0.23 mmol), BINAP (14.00 mg, 0.023 mmol), Pd 2 (dba) 3 (21.00 mg, 0.023 mmol), t-BuONa (43.00 mg, 0.45 mmol) into a two-necked flask. Evacuate with N 2 for 5 minutes, add 3 mL of anhydrous 1,4-dioxane, and stir at 110 °C overnight to obtain Compound 25, 35 mg of a yellow solid with a yield of 28.7%. 11H NMR (300 MHz, DMSO) δ 7.66–7.57 (m, 2H), 7.57–7.47 (m, 3H), 7.46–7.40 (m, 2H), 6.93 (d, J = 8.3 Hz, 2H), 6.84 (d, J = 4.7 Hz, 2H), 6.68 (s, 1H), 5.93 (d, J = 14.0 Hz, 2H), 4.09 (d, J = 15.8 Hz, 1H), 3.74 (s, 3H), 3.73 (s, 3H), 3.27–3.06 (m, 2H), 2.90 (d, J = 12.6 Hz, 1H), 2.63 (d, J = 15.9 Hz, 1H), 2.35 (s, 1H). MS [M+1]: 541.2

[0117] Synthesis of Compound 26 in Example 2 - 7

[0118] Weigh 12 - aminotetrahydroberberine, 1 - tert - butoxycarbonyl - 3 - pyrrolidone, ZnCl 2 , NaBH 3 CN and add them to 3 mL of methanol. Stir at 80 °C and monitor the reaction by TLC until it is complete. Quench with water and extract three times with DCM. Column chromatography with PE:EA = 10:3 gives 1.02 g of a brown - yellow oily liquid. Add 1,4 - dioxane (8 mL) and hydrochloric acid, stir at room temperature for 6 h, adjust the pH to alkaline with 5M NaOH, extract three times with DCM, and evaporate to dryness to obtain 600 mg of a brown - gray solid; Weigh m - bromofluorobenzene (62.00 mg, 0.35 mmol), BINAP (32.00 mg, 0.035 mmol), Pd 2 (dba) 3 (22.00 mg, 0.035 mmol), t - BuONa (67.00 mg, 0.71 mmol), and 150 mg of the above - mentioned brown - gray solid into a two - necked flask. Stir at 80 °C and monitor the reaction by TLC until it is complete. Quench with water and extract three times with DCM. Reverse - phase column chromatography with MeOH:H2O = 35:65~90:10 gives Compound 26, 78 mg of a yellow solid; Yield 44.6%. 11H NMR (400 MHz, chloroform-d) δ 7.18 (p, J = 7.8 Hz, 1H), 6.79–6.69 (m, 1H), 6.62 (s, 1H), 6.47–6.22 (m, 4H), 5.94 (d, J = 3.8 Hz, 2H), 4.36–4.15 (m, 2H), 3.90 (s, 3H), 3.81 (s, 3H), 3.72–3.56 (m, 2H), 3.54–3.38 (m, 3H), 3.33–3.04 (m, 3H), 2.89 (d, J = 15.1 Hz, 1H), 2.70 (s, 2H), 2.49–2.31 (m, 2H), 2.20–1.99 (m, 2H).

[0119] Synthesis of Compound 27 in Example 2 - 8

[0120] Weigh 12 - aminotetrahydroberberine, 1 - tert - butoxycarbonyl - 3 - pyrrolidone, ZnCl 2 , NaBH 3 CN and add them to 3 mL of methanol. Stir at 80 °C and monitor the reaction by TLC until it is complete. Quench the reaction with water and extract three times with DCM. Perform column chromatography with PE:EA = 10:3 to obtain 1.02 g of a pale yellow oily liquid. Add 1,4 - dioxane (8 mL) and hydrochloric acid, stir at room temperature for 6 h, adjust the pH to basic with 5M NaOH, extract three times with DCM, and rotary evaporate to obtain 600 mg of a brown - gray solid; Weigh o - bromofluorobenzene (62.00 mg, 0.35 mmol), BINAP (32.00 mg, 0.035 mmol), Pd 2 (dba) 3 (22.00 mg, 0.035 mmol), t - BuONa (67.00 mg, 0.71 mmol), and 150 mg of the above - mentioned brown - gray solid into a two - necked flask. Stir at 80 °C and monitor the reaction by TLC until it is complete. Quench the reaction with water, extract three times with DCM, and perform reverse - phase column chromatography with MeOH:H2O = 35:65~90:10 to obtain Compound 27, 70 mg of a yellow solid; The yield is 38.8%. 11H NMR (400 MHz, chloroform-d) δ 7.02 (p, J = 6.9 Hz, 2H), 6.78 (s, 1H), 6.77–6.67 (m, 2H), 6.62 (s, 1H), 6.26 (s, 1H), 5.94 (d, J = 3.3 Hz, 2H), 4.29–4.16 (m, 2H), 3.89 (s, 3H), 3.80 (s, 4H), 3.69–3.39 (m, 5H), 3.31–3.05 (m, 3H), 2.90 (dd, J = 15.6, 4.3 Hz, 1H), 2.77–2.59 (m, 2H), 2.40 (dq, J = 13.6, 6.6 Hz, 2H), 2.05 (dq, J = 13.2, 6.4 Hz, 1H). MS [M+1]: 518.1。

[0121] Synthesis of Compound 28 in Example 2-9

[0122] Weigh 12-aminotetrahydroberberine, 1-tert-butoxycarbonyl-3-pyrrolidone, ZnCl 2 , NaBH 3 CN and add them to 3 mL of methanol. Stir at 80 °C and monitor the reaction by TLC until it is complete. Quench with water and extract three times with DCM. Perform column chromatography with PE:EA = 10:3 to obtain 1.02 g of a yellowish-brown oily liquid. Add 1,4-dioxane (8 mL) and hydrochloric acid, stir at room temperature for 6 h, adjust the pH to basic with 5 M NaOH, extract three times with DCM, and evaporate to dryness to obtain 600 mg of a brownish-gray solid. Weigh p-chlorofluorobenzene (62.00 mg, 0.35 mmol), BINAP (32.00 mg, 0.035 mmol), Pd 2 (dba) 3 (22.00 mg, 0.035 mmol), t-BuONa (67.00 mg, 0.71 mmol), and 150 mg of the above brownish-gray solid into a two-necked flask. Stir at 80 °C and monitor the reaction by TLC until it is complete. Quench with water and extract three times with DCM. Perform reverse-phase column chromatography with MeOH:H2O = 35:65~90:10 to obtain Compound 28, 92 mg of a green solid; yield 49.2%. 11H NMR (300 MHz, chloroform-d) δ 7.25–7.14 (m, 2H), 6.73 (d, J = 15.7 Hz, 1H), 6.62 (s, 1H), 6.52 (t, J = 9.3 Hz, 2H), 6.26 (d, J = 2.0 Hz, 1H), 5.94 (dd, J = 2.3, 1.1 Hz, 2H), 4.33–4.18 (m, 2H), 3.90 (s, 3H), 3.81 (s, 3H), 3.74–3.65 (m, 2H), 3.59 (s, 1H), 3.50 (t, J = 7.5 Hz, 1H), 3.38 (tt, J = 9.4, 5.1 Hz, 1H), 3.32–3.05 (m, 3H), 2.88 (s, 1H), 2.70 (s, 2H), 2.41 (tt, J = 14.0, 7.3 Hz, 2H), 2.17–1.98 (m, 2H). MS [M+1]: 534.2。

[0123] Synthesis of Compound 29 in Example 2 - 10

[0124] Weigh 12 - aminotetrahydroberberine, 1 - tert - butoxycarbonyl - 3 - pyrrolidone, ZnCl 2 , NaBH 3 CN and add them to 3 mL of methanol. Stir at 80 °C and monitor the reaction by TLC until it is complete. Quench the reaction with water and extract three times with DCM. Perform column chromatography with PE:EA = 10:3 to obtain 1.02 g of a pale yellow oily liquid. Add 1,4 - dioxane (8 mL) and hydrochloric acid, stir at room temperature for 6 h, adjust the pH to alkaline with 5 M NaOH, extract three times with DCM, and evaporate to dryness to obtain 600 mg of a brown - gray solid. Weigh p - methylfluorobenzene (62.00 mg, 0.35 mmol), BINAP (32.00 mg, 0.035 mmol), Pd 2 (dba) 3 (22.00 mg, 0.035 mmol), t - BuONa (67.00 mg, 0.71 mmol), and 150 mg of the above - mentioned brown - gray solid into a two - necked flask. Stir at 80 °C and monitor the reaction by TLC until it is complete. Quench the reaction with water and extract three times with DCM. Perform reverse - phase column chromatography with MeOH:H2O = 35:65~90:10 to obtain Compound 29, 87 mg of a yellow solid; yield 48.6%. 11H NMR (300 MHz, chloroform-d) δ 7.08 (dd, J = 8.2, 4.3 Hz, 2H), 6.73 (d, J = 17.6 Hz, 1H), 6.62 (s, 1H), 6.54 (t, J = 8.4 Hz, 2H), 6.27 (d, J = 2.3 Hz, 1H), 5.99–5.89 (m, 2H), 4.29–4.18 (m, 2H), 3.89 (s, 3H), 3.81 (s, 3H), 3.75–3.55 (m, 3H), 3.55–3.42 (m, 2H), 3.41–3.27 (m, 2H), 3.14 (dd, J = 9.7, 4.2 Hz, 1H), 2.89 (d, J = 14.9 Hz, 1H), 2.69 (s, 2H), 2.49–2.30 (m, 2H), 2.30–2.24 (m, 3H), 2.09 (qd, J = 11.4, 5.4 Hz, 2H). MS [M+1]: 514.3

[0125] Synthesis of Compound 30 in Example 2-11

[0126] Weigh intermediate 4 (100.00 mg, 0.28 mmol), 2-bromo-6-fluoronaphthalene (63.00 mg, 0.28 mmol), BINAP (18.00 mg, 0.028 mmol), Pd 2 (dba) 3 (26.00 mg, 0.028 mmol), and sodium tert-butoxide (54.00 mg, 0.56 mmol) into a two-necked flask. Add 3 mL of anhydrous 1,4-dioxane and stir at 110 °C for 12 h. After monitoring the reaction to completion by TLC, quench with water, extract three times with DCM, and perform column chromatography with PE:EA = 10:3 to obtain 78 mg of a pale yellow solid with a yield of 56%. 1 1H NMR (300 MHz, DMSO) δ 7.78–7.65 (m, 2H), 7.61–7.48 (m, 2H), 7.33–7.20 (m, 2H), 7.15 (d, J = 2.2 Hz, 1H), 6.87 (s, 1H), 6.83 (s, 1H), 6.66 (s, 1H), 5.92 (d, J = 14.9 Hz, 2H), 4.10 (d, J = 15.8 Hz, 1H), 3.74 (d, J = 2.3 Hz, 6H), 3.43 (d, J = 15.1 Hz, 2H), 3.26 (d, J = 16.5 Hz, 1H), 3.12 (d, J = 10.3 Hz, 1H), 2.89 (d, J = 11.3 Hz, 1H), 2.61 (d, J = 15.8 Hz, 1H), 2.48–2.25 (m, 2H). MS [M+1]: 499.2

[0127] Example 2-12 Synthesis of Compound 31

[0128] Weigh p-fluorophenylboric acid (200.00 mg, 1.43 mmol), p-dibromobenzene (300.00 mg, 1.43 mmol), Pd(PPh 3 ) 4 (165 mg, 0.143 mmol), Cs 2 CO 3 (931 mg, 1.43 mmol) was placed in a two-necked bottle, N 2 After evacuation for 5 minutes, 8 mL of 1,4-dioxane was added and stirred at 110°C for 6 h. After the reaction was completed, water was added to quench the reaction, and DCM was extracted three times. PE column chromatography was performed to obtain 194 mg of a white solid with a yield of 57.1%. The above white solid (57.00 mg, 0.23 mmol), 12-aminotetrahydroberberine (80.00 mg, 0.23 mmol), BINAP (14.00 mg, 0.023 mmol), Pd 2 (dba) 3 (21.00 mg, 0.023 mmol), t-BuONa (43.00 mg, 0.45 mmol) were placed in a two-necked bottle, N 2 The mixture was evacuated for 5 minutes, 3 mL of anhydrous 1,4-dioxane was added, and the mixture was stirred at 110° C. overnight to obtain 54 mg of a yellow solid with a yield of 44.8%. 1 H NMR (300MHz, DMSO) δ7.61 (dd, J=8.6, 5.5Hz, 2H), 7.47 (d, J=8.8Hz, 3H), 7.22 (t, J=8.7Hz, 2 H),6.92(d,J=8.3Hz,2H),6.83(d,J=8.1Hz,2H),6.67(s,1H),5.93(d,J=14.0Hz,2H),4.08( d,J=15.9Hz,1H),3.73(d,J=3.3Hz,6H),3.44(s,2H),3.21(d,J=3.7Hz,1H),3.12(d,J=10. 6Hz,1H),2.90(d,J=13.1Hz,1H),2.62(d,J=15.8Hz,1H),2.44–2.25(m,2H).MS[M+1]:525.2

[0129] Example 3

[0130] The reaction formula of the compound of structural formula 2 is as follows:

[0131]

[0132] Synthesis of intermediate 5

[0133] Under an ice-salt bath, intermediate 4 (800.00 mg, 2.26 mmol) was added, and then dilute H 2 SO 4 (20%, 8.00 mL) was added. After stirring for 5 min until completely mixed, an aqueous solution of 40% mass fraction of NaNO 2 (187.10 mg, 2.71 mmol) was gradually added dropwise, and the reaction was carried out for half an hour. After half an hour, KI (562.70 mg, 3.39 mmol) was dissolved in 2 mL of water and added dropwise to the above reaction solution, and the reaction was carried out under an ice bath for 4 hours. Subsequently, an aqueous NaOH solution was added to neutralize to alkalinity, and extraction was carried out three times with DCM. The organic phases were combined, dried over anhydrous sodium sulfate, and subjected to column chromatography with PE:EA = 10:1 to obtain 12-iodotetrahydroberberine, 420.00 mg of yellow solid, and the yield was 40.0%. 1 H NMR (300 MHz, DMSO) δ 7.39 (s, 1H), 6.92 (s, 1H), 6.69 (s, 1H), 5.97 (dd, J = 9.5, 1.0 Hz, 2H), 4.04 (d, J = 15.9 Hz, 1H), 3.80 (s, 3H), 3.73 (s, 3H), 3.40 (d, J = 15.3 Hz, 2H), 3.17–3.06 (m, 2H), 2.90 (t, J = 13.7 Hz, 1H), 2.62 (d, J = 15.9 Hz, 1H), 2.47–2.40 (m, 1H), 2.29 (dd, J = 16.2, 10.9 Hz, 1H).

[0134] Synthesis of Compound 32 in Example 3-1

[0135] Intermediate 5 (80.00 mg, 0.17 mmol), 4-chlorophenylboronic acid (32 mg, 0.21 mmol), cesium carbonate (56.00 mg, 0.17 mmol), and tetrakis(triphenylphosphine)palladium (9.94 mg, 0.0086 mmol) were placed in a two-necked flask, and 4 mL of a mixed solvent of anhydrous 1,4-dioxane:H2O = 3:1 was added. The mixture was evacuated with nitrogen for 5 min and reacted at 80 °C. The reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted three times with DCM, the organic phases were combined, dried over anhydrous sodium sulfate, and subjected to column chromatography with PE:EA = 1:1 to obtain 25 mg of yellow solid, and the yield was 32.4%. 11H NMR (300 MHz, DMSO) δ 7.48 (d, J = 8.5 Hz, 2H), 7.40 (d, J = 8.4 Hz, 2H), 6.79 (s, 1H), 6.65 (d, J = 9.0 Hz, 2H), 5.91 (d, J = 16.0 Hz, 2H), 4.13 (d, J = 5.2 Hz, 1H), 3.81 (s, 3H), 3.78 (s, 3H), 3.47 (d, J = 15.9 Hz, 1H), 3.11 (d, J = 9.8 Hz, 1H), 2.88 (d, J = 15.7 Hz, 2H), 2.60 (d, J = 15.2 Hz, 2H), 2.43 (d, J = 11.1 Hz, 1H). MS [M+1]: 430.2

[0136] Synthesis of Compound 33 in Example 3-2

[0137] Intermediate 5 (70 mg, 0.15 mmol), p-tolylboronic acid (25 mg, 0.18 mmol), cesium carbonate (49 mg, 0.15 mmol), and tetrakis(triphenylphosphine)palladium(0) (9.00 mg, 0.0075 mmol) were placed in a two-necked flask. 4 mL of a mixed solvent of anhydrous 1,4-dioxane:H2O = 3:1 was added, and the mixture was evacuated with nitrogen for 5 min and reacted at 80 °C. The reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted three times with DCM, the organic phases were combined, dried over anhydrous sodium sulfate, and purified by column chromatography (PE:EA = 1:1) to obtain 28 mg of a yellow solid with a yield of 43.3%. 1 1H NMR (300 MHz, DMSO) δ 7.24 (d, J = 1.1 Hz, 4H), 6.75 (s, 1H), 6.66 (s, 1H), 6.57 (s, 1H), 5.91 (dd, J = 15.5, 1.0 Hz, 2H), 4.13 (d, J = 16.0 Hz, 1H), 3.80 (s, 3H), 3.77 (s, 3H), 3.52–3.41 (m, 1H), 3.30 (s, 1H), 3.19–3.07 (m, 1H), 2.97–2.83 (m, 2H), 2.60 (d, J = 14.8 Hz, 2H), 2.48–2.39 (m, 1H), 2.35 (s, 3H). MS [M+1]: 450.1

[0138] Synthesis of Compound 34 in Example 3-3

[0139] Intermediate 5 (70 mg, 0.15 mmol), p-fluorophenylboronic acid (25 mg, 0.18 mmol), cesium carbonate (49 mg, 0.15 mmol), and tetrakis(triphenylphosphine)palladium (9.00 mg, 0.0075 mmol) were placed in a two-necked flask. 4 mL of a mixed solvent of anhydrous 1,4-dioxane:H2O = 3:1 was added. The mixture was evacuated with nitrogen for 5 min and reacted at 80 °C. The reaction was monitored by TLC. After the reaction was complete, water was added to quench the reaction. The mixture was extracted three times with DCM. The organic phases were combined, dried over anhydrous sodium sulfate, and purified by column chromatography (PE:EA = 1:1) to obtain 40 mg of a yellow solid with a yield of 61.4%. 1 1H NMR (400 MHz, Chloroform-d) δ 7.39–7.33 (m, 2H), 7.09–7.02 (m, 2H), 6.76 (s, 1H), 6.62 (s, 1H), 6.15 (s, 1H), 5.93 (q, J = 1.5 Hz, 2H), 4.35 (s, 2H), 4.25 (d, J = 15.7 Hz, 1H), 3.79 (s, 3H), 3.78 (s, 3H), 3.66–3.54 (m, 2H), 3.29–3.21 (m, 1H), 3.14 (ddd, J = 15.0, 10.4, 4.6 Hz, 1H), 2.95 (ddd, J = 15.3, 4.4, 1.2 Hz, 1H), 2.74–2.61 (m, 2H), 2.44 (ddd, J = 15.3, 11.0, 1.5 Hz, 1H). MS [M+1]: 434.2

[0140] Synthesis of Compound 35 in Example 4

[0141] The reaction formula of the compound of Structural Formula 3 is as follows:

[0142]

[0143] Intermediate 4 (80.00 mg, 0.23 mmol) was weighed and dissolved in THF (3 mL). Under an ice bath, Et3N (68.60 mg, 94 μL, 0.68 mmol) was added dropwise and stirred for 2 min. After mixing, 4-fluorobenzoyl chloride (36.00 mg, 0.23 mmol) was added dropwise. When the reaction was monitored by TLC and completed, water was added to quench the reaction. The mixture was extracted three times with DCM. The organic phases were combined, dried over anhydrous sodium sulfate, and purified by column chromatography (PE:EA = 1:1) to obtain 54 mg of a yellow solid with a yield of 50.2%. 11H NMR (300 MHz, DMSO) δ 9.78 (s, 1H), 8.05 (dd, J = 8.7, 5.5 Hz, 2H), 7.37 (t, J = 8.9 Hz, 2H), 7.01 (s, 1H), 6.82 (s, 1H), 6.68 (s, 1H), 5.92 (dd, J = 14.5, 1.1 Hz, 2H), 4.09 (d, J = 15.9 Hz, 1H), 3.79 (s, 3H), 3.76 (s, 3H), 3.22–3.07 (m, 2H), 2.99–2.84 (m, 2H), 2.61 (d, J = 15.7 Hz, 2H). MS [M+1]: 477.2

[0144] Example 5 Pharmacology Part

[0145] 1. PI Detection

[0146] PI (Propidium Iodide) is a nuclear dye that can penetrate the cell membrane that has lost integrity and bind to nucleic acids to emit red fluorescence. PI cannot enter living cells because their cell membranes are intact. In dead cells, the integrity of the cell membrane and nuclear membrane is reduced, allowing PI to enter the cells and stain. Therefore, PI can be used as an indicator to distinguish between living and dead cells. HK-2 cells (cell source: purchased from the Chinese Cell Bank) in the logarithmic growth phase were seeded in 6-well plates at 300,000 cells per well. The next day, when the cell density reached about 60%, the drug was administered. After 24 h of drug administration, the supernatant was collected, and then the cells were washed twice with cold PBS, digested with 1 ml of trypsin, collected, and centrifuged. Subsequently, 300 μL of 1×Binding Buffer was added to each sample, and then 2.5 μL of PI dye was added. After mixing and resuspending the cells, they were incubated in the dark at room temperature for 10 min, and then filtered and analyzed by flow cytometry.

[0147] Table 4 PI Measurement Values of Compounds 1 - 13

[0148]

[0149]

[0150] 2. Cell Viability Assay

[0151] HK-2 cells (cell source: purchased from the Chinese Cell Bank) were seeded into 96-well plates and treated with the drug and Erastin for 24 h. After treatment, the medium was replaced with fresh medium (DMEM / F12 medium with 10% FBS (KeyGen Biotech, Nanjing, China)), and 10 μl of CCK-8 was added to each well. The 96-well plates were incubated at 37 °C for 1 h, and the absorbance at 450 nm was measured using a microplate reader. The absorbance is proportional to the cell survival rate.

[0152] EC50 and E of Some Compounds in Table 5 max Value

[0153]

[0154] a The EC50 values are expressed as the mean ± standard deviation of repeated measurements. b Emax is the highest value of cell viability calculated from the EC50 curve (the maximum concentration is 500 nM).

[0155] 2. Detection of Lipid Peroxidation

[0156] BODIPY 581 / 591C11 is a fluorescence ratio probe for lipid oxidation, which can be used to detect reactive oxygen species (ROS) in cells and cell membranes, and is usually used to indicate the level of lipid peroxidation in living cells. HK-2 cells in the logarithmic growth phase were seeded in 6-well plates at 300,000 cells per well. The next day, when the cell density reached about 60%, the drugs were administered. After 24 h of drug administration, the supernatant was collected, and the cells were washed twice with cold PBS, digested with 1 ml of trypsin, collected and centrifuged. A certain amount of 10 mM C11 BODIPY 581 / 591 stock solution was added to make the final concentration reach 10 μM. Incubate at 37 °C for 30 min. The fluorescence reading ratio measured at the emission wavelengths of 590 nm / 510 nm reflects the level of cell lipid peroxidation.

[0157] The results are as Figure 1 shown. It can be seen from the figure that Compounds 20, 21, 30, and 31 can effectively reduce the ROS values in Erastin-treated HK-2 cells at concentrations of 5 nM, 10 nM, and 20 nM.

[0158] 3. Detection of MDA

[0159] Malondialdehyde (MDA) is a natural product of lipid oxidation in organisms. When oxidative stress occurs in animal or plant cells, lipid oxidation occurs. Some fatty acids are gradually decomposed into a series of complex compounds after oxidation, including MDA. The reaction of MDA with thiobarbituric acid (TBA) can produce a color reaction of a red product, and then colorimetry is used for quantitative detection of MDA in cell lysates. HK-2 cells after drug treatment for a specific time were collected with a cell scraper, rinsed once with PBS, and the liquid was collected and centrifuged at 2500 rpm for 5 min, and the supernatant was discarded.

[0160] Total protein extraction: Resuspend the collected cells with pre-cooled PBS, transfer them into a 1.5 ml tube, centrifuge at 2500 rpm for 5 min, discard the supernatant, centrifuge at 15000 rpm for 30 s, and aspirate the residual liquid with a pipette. Add 10 μL of protease inhibitor (purchased from TargetMol) and 10 μL of phosphatase inhibitor to 980 μL of RIPA buffer, add an appropriate amount of RIPA lysis buffer according to the cell pellet and mix well, place on ice for lysis, vortex vigorously every 10 minutes. After 30 min, centrifuge at 4 °C and 15000 rpm for 20 min, collect the supernatant, and take 1 μL of the sample to detect the protein concentration by the BCA method. Add 0.1 ml of lysis buffer into a centrifuge tube or other appropriate container as a blank control, add 0.1 ml of the above different concentration standards to make a standard curve, and add 0.1 ml of the sample for determination; then add 0.2 ml of MDA detection working solution. After mixing, heat at 100 °C or in a boiling water bath for 15 minutes.

[0161] Cool to room temperature in a water bath and centrifuge at 1000 g for 10 minutes at room temperature. Take 200 μL of the supernatant and add it to a 96-well plate, and then measure the absorbance at 532 nm with an enzyme-linked immunosorbent assay (ELISA) reader. Calculation of MDA content: For cell samples, after calculating the MDA content in the sample solution, the MDA content in the original sample can be expressed by the protein content or tissue weight per unit weight, etc.

[0162] The results are as Figure 2 shown. It can be seen from the figure that compounds 20, 21, 30, and 31 can effectively reduce the MDA value in HK-2 cells after Erastin treatment at concentrations of 5 nM, 10 nM, and 20 nM.

[0163] 4. Renal injury model

[0164] Animal model: Intraperitoneally inject a folic acid solution at 200 mg / kg, then administer the drug by gavage. After 72 h, sacrifice the mice, collect the serum and kidneys. Fix one kidney in PUFA for 24 hours, embed it in paraffin and section it, and perform HE staining and PAS staining. Freeze and thaw the other kidney repeatedly in liquid nitrogen, then perform tissue homogenization, extract RNA, and perform QPCR (Real-time Quantitative PCR Detecting System).

[0165] The results are as Figure 3 shown. It can be seen from the figure that compounds 20, 21, 30, and 31 can effectively treat acute kidney injury induced by folic acid.

[0166] The results are as Figure 4As shown, it can be seen from the figure that compounds 20, 21, 30, and 31 can effectively reduce the mRNA expression of kidney injury molecule-1 (KIM-1) in mice with acute kidney injury.

[0167] The results are as Figure 5 shown. It can be seen from the figure that compounds 20, 21, 30, and 31 can effectively reduce the mRNA expression of neutrophil gelatinase-associated lipocalin (NGAL) in mice with acute kidney injury.

[0168] 5. Serum creatinine detection

[0169] Creatinine (CRE) is a product of muscle metabolism and is mainly excreted through glomerular filtration. Under normal circumstances, the content of creatinine in the body is basically stable. The creatinine concentration in the blood can be used as one of the indicators for detecting glomerular filtration function. Creatinine is specifically acted on by creatinine enzyme to generate creatine, and creatine is successively acted on by creatine kinase and sarcosine oxidase to generate hydrogen peroxide. Hydrogen peroxide reacts with the chromogenic agent to present a purple color. This colored substance has a large absorption peak at 546 nm, and then the creatinine content is calculated.

[0170] Sample preparation: Tissue sample: Take about 0.1 g of tissue sample, grind it with 1 mL of extraction solution, transfer all the crude extract to an EP tube, centrifuge at 12000 rpm at room temperature for 10 min, and the supernatant is to be measured. Liquid sample: Clear liquid can be directly detected; if it is turbid, centrifuge and take the supernatant for detection. Then detect it on the machine.

[0171] The results are as Figure 6 shown. It can be seen from the figure that compounds 20, 21, 30, and 31 can effectively reduce the content of serum creatinine (SCR) in mice with acute kidney injury.

[0172] 6. Urea nitrogen detection

[0173] Urea is the end product of the decomposition of nitrogen-containing compounds in organisms and is decomposed and converted into ammonia under the catalysis of urease. Blood urea nitrogen is one of the main indicators of renal function. Urea nitrogen in the sample is co-boiled with diacetyl monoxime and thiosemicarbazide in a ferric chloride-phosphoric acid solution to form a red diazine compound. The depth of its color is proportional to the content of urea nitrogen. The diacetyl monoxime method is used to determine the urea nitrogen content.

[0174] Sample treatment:

[0175] 1. Tissue: Add distilled water according to the ratio of mass (g): volume of distilled water (mL) of 1:5 - 10 (it is recommended to weigh about 0.1 g and add 1 mL of distilled water), homogenize, centrifuge at 25 °C and 10000 g for 10 min, and take the supernatant for measurement.

[0176] 2. Cells: According to the ratio of the number of cells (104 cells): the volume of distilled water (mL) of 500 - 1000:1 (it is recommended to add 1 mL of distilled water to 5 million cells), break the cells by ultrasonic wave in an ice bath (power 300 w, ultrasonic wave for 3 seconds, interval for 7 seconds, total time 3 min); then centrifuge at 4°C and 10000 g for 10 min, take the supernatant and place it on ice for further measurement.

[0177] 3. Serum or other liquids: Detect directly.

[0178] The results are as Figure 7 shown. It can be seen from the figure that compounds 20, 21, 30, and 31 can effectively reduce the content of blood urea nitrogen (BUN) in mice with acute kidney injury.

[0179] In summary, after the modification of THB, a series of related derivatives have been obtained, and the cell activities of most compounds have been significantly improved compared with THB. Among them, the EC50 values of compounds 20, 21, 30, and 31 have been improved by about 100 times compared with the activity of THB, showing a significant improvement. Moreover, compounds 20, 21, 30, and 31 also show therapeutic effects in the mouse model of acute kidney injury, and can effectively reduce the contents of KIM-1, NGAL, SCR, and BUN. In addition, the tissue analysis of mice with acute kidney injury also shows that compounds 20, 21, 30, and 31 have a protective effect on the kidneys.

[0180] The above schematically describes the present invention and its implementation manners. This description is not restrictive. What is shown in the drawings is only one of the implementation manners of the present invention, and the actual structure is not limited thereto. Therefore, if those of ordinary skill in the art are inspired by it and design similar structural manners and embodiments without creative efforts without departing from the purpose of the present invention, they shall fall within the protection scope of the present invention.

Claims

1. A tetrahydroberberine derivative, characterized in that: The general formula of the tetrahydroberberine derivatives is as follows (I): As shown, Wherein, when X=N or O, Y=(CH2)n, n=0-6, R=aromatic ring, aromatic heterocycle, monosubstituted or polysubstituted aromatic ring and aromatic heterocycle, specifically as As shown; When X=Y=0, R=aromatic ring and its mono-substituted or poly-substituted aromatic ring, aromatic heterocycle and its mono-substituted or poly-substituted aromatic heterocycle, specifically As shown; When X=N,O, When R = an aromatic ring, an aromatic heterocycle, a monosubstituted or polysubstituted aromatic ring or an aromatic heterocycle, specifically shown.

2. The tetrahydroberberine derivative according to claim 1, characterized in that: When X=Y=0, R is p-chlorophenyl, p-fluorophenyl or p-methylphenyl.

3. The tetrahydroberberine derivative according to claim 1, characterized in that: When X=N, O, Y=(CH2)n, n=0-3 or a four-membered ring, five-membered ring or aromatic ring containing heteroatoms, R is p-chlorophenyl, p-fluorophenyl, p-methylphenyl, phenyl, p-hydroxyphenyl, p-fluoropyridine, 1,2,4-trifluorophenyl, 4-hydroxymethylfuranyl, 4-fluoro-2-chlorophenyl, 2,4-dichlorophenyl, 2,4,6-trifluorophenyl, 2,6-difluoro-4-hydroxyphenyl, 4-acetamidophenyl or 4-cyanophenyl.

4. The tetrahydroberberine derivative according to claim 1, characterized in that: When X=N,O, When R is p-fluorophenyl.

5. The tetrahydroberberine derivative according to claim 1, characterized in that: The salt is selected from the group consisting of hydrobromide, hydroiodide, hydrofluoride, hydrochloride, sulfate, nitrate, phosphate, citrate, acetate, and lactate.

6. The tetrahydroberberine derivative according to claim 5, characterized in that: The salt is hydrobromide, hydrochloride or sulfate.

7. The tetrahydroberberine derivative according to claim 1, characterized in that: The tetrahydroberberine derivatives are specifically 12-(N-(4-hydroxybenzyl))aminotetrahydroberberine; 12-(N-(2,4-dihydroxybenzyl))aminotetrahydroberberine; 12-(N-(4-acetylaminobenzyl))aminotetrahydroberberine; 12-(N-(3-hydroxymethylfuran))aminotetrahydroberberine; 12-(N-(2,4-dichlorobenzyl))aminotetrahydroberberine; 12-(N-(2,4,6-trifluorobenzyl))aminotetrahydroberberine; 12-(N-(4-hydroxy-2,6-difluorobenzyl))aminotetrahydroberberine; 12-(N-(4-fluorobenzyl))aminotetrahydroberberine; 12-(N-( 2,4-difluorobenzyl)) aminotetrahydroberberine; 12-(N-(2,4,5-trifluorobenzyl)) aminotetrahydroberberine; 12-(N-(2-dichloro-4-fluoro-benzyl)) aminotetrahydroberberine; 12-(N-(2-fluoropyridin-5-methylene-)) aminotetrahydroberberine; 12-(N-(4-cyanobenzyl)) aminotetrahydroberberine; 12-(N-benzyl) aminotetrahydroberberine; 12-(N-(4-chlorobenzyl)) aminotetrahydroberberine; 12-(N-(4-methylbenzyl)) aminotetrahydroberberine; 12-(N-methyl, N-(4-fluorobenzyl)) aminotetrahydroberberine; 12-(N-(2-( 4-fluorophenyl)ethyl))aminotetrahydroberberine; 12-(N-(2-(4-fluorophenyl)propyl))aminotetrahydroberberine; 12-(N-(4-fluorophenyl))aminotetrahydroberberine; N-(4'-fluoro-1,1'-biphenyl-3-yl)aminotetrahydroberberine; 12-(N-(1-(p-fluorophenyl)pyrrolidin-3-yl))aminotetrahydroberberine; N-(6-(4-fluorophenyl)pyridin-2-yl)aminotetrahydroberberine; N-(4'-methyl-1,1'-biphenyl-4-yl)aminotetrahydroberberine; N-(4'-chloro-1,1'-biphenyl-4-yl)aminotetrahydroberberine; 12-(N-(1-(p-fluorophenyl)pyrrolidin-3-yl))aminotetrahydroberberine 12-(N-(1-(p-fluorophenyl)pyrrolidin-3-yl))aminotetrahydroberberine; 12-(N-(1-(p-chlorophenyl)pyrrolidin-3-yl))aminotetrahydroberberine; 12-(N-(1-(p-methylphenyl)pyrrolidin-3-yl))aminotetrahydroberberine; N-(6-fluoronaphthalen-2-yl)aminotetrahydroberberine; N-(4'-methyl-1,1'-biphenyl-4-yl)aminotetrahydroberberine; 12-p-chlorophenyltetrahydroberberine; 12-p-methylphenyltetrahydroberberine; 12-p-fluorophenyltetrahydroberberine; 12-(p-fluorobenzamide)tetrahydroberberine.

8. An inhibitor, characterized in that The active ingredient of the inhibitor is the tetrahydroberberine derivative according to any one of claims 1 to 7.

9. A pharmaceutical composition comprising the tetrahydroberberine derivative according to any one of claims 1 to 7.

10. Use of the tetrahydroberberine derivatives according to any one of claims 1 to 7, the inhibitor according to claim 8 or the pharmaceutical composition according to claim 9 in the preparation of medicines for preventing, alleviating and / or treating ferroptosis-related diseases.

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