Pyridone compounds, synthesis method, and use thereof

Abstract

Disclosed in the present invention are pyridone derivatives, having structures represented by general formula I, general formula II or general formula III. Also disclosed in the present invention is a preparation method for the described derivatives, etc. Studies have shown that the resistance indexes (RI=IC50[I38T] / IC50[WT]) of the hydroxypyridone derivatives described in the present invention against wild-type and PA / I38T mutation-type influenza virus RNA polymerases are lower than that of baloxavir, which indicates that the derivatives of the present invention are less affected by PA / I38T mutation-type influenza A viruses and are superior to baloxavir. These derivatives can also effectively inhibit virus replication of avian influenza H5N1, and have lower cytotoxicity than baloxavir and have higher selectivity, and therefore can be used for preparing anti-influenza drugs for inhibiting baloxavir-resistant virus strains and avian influenza.

Description

Pyridinone compound, synthetic method and application thereof TECHNICAL FIELD

[0001] The present application belongs to the field of pharmaceutical chemistry, and particularly relates to a pyridinone compound, a synthetic method and application thereof. BACKGROUND

[0002] Influenza spreads rapidly in the population, and has a high incidence and mortality rate. Anti-viral treatment of influenza mainly relies on neuraminidase inhibitors, but drug resistance has already occurred, and new anti-viral drugs of new mechanisms are urgently needed. In 2018, the influenza virus cap-dependent endonuclease inhibitor baloxavir marboxil was approved for the treatment of acute uncomplicated influenza, and has a completely new mechanism of action. However, in recent clinical studies, baloxavir has produced a variety of different mutant virus strains. Among them, the PA / I38X (I38T / M / F / L / N / S mutation) mutation accounts for as much as 2%-9% in patients. The conventional dose of baloxavir is difficult to produce the expected therapeutic effect on influenza patients carrying the I38X mutation. It is urgent to develop PA inhibitors with the same level of inhibitory activity against wild-type and mutant influenza virus strains. The reduced sensitivity of BXA to the I38T mutation is attributed to unfavorable van der Waals packing and induced adaptive changes of the Thr38 residue. The butterfly-shaped hydrophobic fragment of BXA provides a nearly perfect filling of the hydrophobic pocket of the active site. However, the effect of this filling is slightly reduced with the appearance of Thr38, which is more polar and smaller in size. In view of the change in the binding mode caused by the point mutation of the hydrophobic pocket of the active site of PA, the development of a new type of hydrophobic fragment substituent to further weaken the interaction between the corresponding inhibitor and the I38T mutation site is expected to improve its inhibitory activity against the influenza PA / I38T mutant strain. N Active site hydrophobic pocket provides a nearly perfect filling. However, the effect of this filling is slightly reduced with the appearance of Thr38, which is more polar and smaller in size. In view of the change in the binding mode caused by the point mutation of the hydrophobic pocket of the active site of PA N Active site hydrophobic pocket provides a nearly perfect filling. However, the effect of this filling is slightly reduced with the appearance of Thr38, which is more polar and smaller in size. In view of the change in the binding mode caused by the point mutation of the hydrophobic pocket of the active site of PA SUMMARY

[0003] The present application aims to provide a pyridinone derivative, which is a hydroxypyridinone derivative having 5' cap-like structure (CAP) dependent endonuclease inhibitory activity.

[0004] Another object of the present application is to provide a pyridinone derivative capable of simultaneously inhibiting PA / WT and PA / I38T type influenza viruses.

[0005] The present application further provides a use of the compound in the preparation of an anti-influenza virus drug for inhibiting the activity of influenza virus RNA polymerase.

[0006] Specifically, the following formula (I), formula (II) and formula (III) shown in the structure or a pharmaceutically acceptable salt thereof,

[0007] wherein

[0008] R1 is selected from the group consisting of hydrogen, halogen, alkyl (preferably C1-C6 alkyl), R1 is one or independently multiple;

[0009] R2 is selected from the group consisting of nothing, alkyl (preferably C1-C6 alkyl) or alkanoyl (preferably C1-C3 alkanoyl, further preferably

[0010] R3 is selected from the group consisting of hydrogen or

[0011] R4 is selected from the group consisting of hydrogen or alkyl (preferably C1-C6 alkyl);

[0012] X is selected from the group consisting of N, O, S, Se; when X is O, S, Se, R2 is nothing;

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

[0014] In the present application, the alkyl mentioned is generally C1-C6 alkyl, including but not limited to methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, pentyl, hexyl and the like. The alkyl is further preferably C1-C5 alkyl, and further preferably C1-C3 alkyl, including but not limited to methyl, ethyl, propyl.

[0015] In the present application, the halogen mentioned includes F, Cl, Br, I, as preferred, the halogen includes F, Cl, Br.

[0016] Further, the corresponding C atom in the following formula (I), formula (II) and formula (III) is S configuration or R configuration or a mixture containing S configuration and R configuration:

[0017] R1, R2, R3, R4 and X are defined as above.

[0018] Further, R1 is one, two or independently three, each R1 is independently selected from the group consisting of hydrogen, F, Cl, methyl, ethyl.

[0019] Further, R2 is selected from the group consisting of nothing, or

[0020] Further, R3 is selected from the group consisting of hydrogen or

[0021] Further, R4 is selected from the group consisting of hydrogen or methyl, ethyl;

[0022] Further, X is selected from the group consisting of N or O or S or Se; when X is O, S, Se, R2 is nothing.

[0023] Further, the derivatives of pyridinone have the following formula (I-a), (I-b), (I-c), (III-a) structure:

[0024] R1is H, halogen, C1-C3alkyl, R1is one or two independently, R3is H or

[0025] Further, the derivatives of pyridinone have the following formula (I-a), (I-b), (I-c), (III-a) structure:

[0026] An intermediate for preparing the derivatives of pyridinone of any of the above technical solutions has the following formula structure:

[0027] R1is selected from hydrogen, halogen, alkyl (preferably C1-C6alkyl), R1is one or multiple independently;

[0028] R2is selected from nothing, alkyl (preferably C1-C6alkyl) or alkanoyl (preferably C1-C3alkyl, further preferably )

[0029] R4is selected from hydrogen or (preferably C1-C6alkyl) alkyl;

[0030] X is selected from N or O or S or Se; when X is O or S or Se, R2is nothing;

[0031] Ring A is selected from one of the following structures:

[0032] Further, the intermediate has the following formula structure:

[0033] A method for synthesizing a pyridone derivative according to any of the above technical solutions, comprising: reducing the compound shown in formula (I-1), (II-1), or (III-1) in the presence of sodium borohydride to obtain the intermediate shown in formula (I-2), (II-2), or (III-2); reacting the intermediate (I-2), (II-2), or (III-2) with (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H-[1,4]hydrazine[3,4-c]pyrido[2,1-f][1,2,,4]triazine-6,8-dione in the presence of n-butylphosphine anhydride to obtain the intermediate (I-3), (II-3), or (III-3); and reducing the intermediate (I-3), (II-3), or (III-3) with LiCl to obtain the pyridone derivative shown in formula (I), (II), or (III).

[0034] When preparing the intermediate of formula (I-2), (II-2), or (III-2), the solvent used is methanol or ethanol, and the molar ratio of the compound of formula (I-1), (II-1), or (III-1) to sodium borohydride is 1:1 to 2.5.

[0035] When preparing intermediates (I-3), (II-3), or formula (III-3), ethyl acetate is used as the solvent; the reaction temperature is 70–120 °C; and the further reaction temperature is 100–120 °C. The molar ratio of intermediate (I-2), (II-2), or formula (III-2) to n-butylphosphine anhydride and (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H-[1,4]hydrazine[3,4-c]pyrido[2,1-f][1,2,4]triazine-6,8-dione is 1:1–1.5:0.3–0.6.

[0036] When preparing the pyridone derivatives of formula (I), (II), or (III), the solvent used is N,N-dimethylacetamide; the reaction temperature is 60–100 °C; more preferably 70–90 °C; the molar ratio of intermediate (I-3) or (II-3) of formula (III-3) to LiCl is 1:2–15; even more preferably 1:5–12.

[0037] Furthermore, Ring A For or At this time, the intermediate shown in formula (II-2) first reacts with PBr3 to give intermediate (II-2-a):

[0038] Then, the intermediate (II-2-a) is reacted with (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H- [1,4]azepino[3,4-c]pyrido[2,1-f][1,2,4]triazine-6,8-dione in the presence of CsF to obtain the intermediate of formula (II-3).

[0039] In the above step, when the intermediate (II-2-a) is prepared, the intermediate of formula (II-2) is first reacted with PBr3 at a molar ratio of 1:1.2-2, and the reaction solvent is anhydrous DCM.

[0040] When the intermediate (II-3) is prepared, the reaction solvent is anhydrous acetonitrile, and the molar ratio of (II-2-a) to Cs to (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H- [1,4]azepino[3,4-c]pyrido[2,1-f][1,2,4]triazine-6,8-dione is 1:1.5-2.5:0.7-1.2, and the reaction temperature is 50-80°C.

[0041] Further:

[0042] When the intermediate (I-1) is of formula (I-1-1), the intermediate (I-1) is obtained by reacting intermediate (I-1-1-1) with tert-butyl nitrite; as preferred, the reaction conditions of this step are: the reaction temperature is 70-100°C, the reaction solvent is DMSO, etc., and the molar ratio of intermediate (I-1-1-1) to tert-butyl nitrite is 1:1.5-4.

[0043] When the intermediate (I-1) is of formula (I-1-2), the intermediate (I-1) is obtained by reacting intermediate (I-1-2-1) with tert-butyl nitrite:

[0044] When the intermediate (I-1) is of formula (I-1-3), the intermediate (I-1) is obtained by reacting intermediate (I-1-3-1) in the presence of polyphosphoric acid:

[0045] As preferred, the reaction conditions of this step are: the reaction temperature is 70-100°C, the reaction solvent is DMSO, etc., and the molar ratio of intermediate (I-1-2-1) to tert-butyl nitrite is 1:1.5-4.

[0046] When the intermediate (II-1) is of formula (II-1-1), the intermediate (II-1) is obtained by reacting intermediate (II-1-1-1) with phenylacetylene and tert-butyl nitrite:

[0047] As preferred, the reaction conditions of this step are as follows: the reaction temperature is 50-80°C, the reaction solvent is chloroform or the like; sodium acetate is added simultaneously; the molar ratio of intermediate (II-1-1), phenylacetylene, sodium acetate and tert-butyl nitrite is 1:1-1.5:1-1.5:1.5-4.

[0048] When the intermediate (II-1) has the structure shown in formula (II-1-2), the intermediate (II-1) is obtained by reacting intermediate (II-1-2-1) with the corresponding Grignard reagent of formula (II-1-2-2) under the action of I:

[0049] As preferred, the reaction conditions of this step are as follows: the reaction solvent is anhydrous THF or the like; the molar ratio of intermediate (II-1-2-1) and the corresponding Grignard reagent of formula (II-1-2-2) is 1:1-2.

[0050] When the intermediate (III-1) has the structure shown in formula (III-1-1), the intermediate (III-1) is obtained by first reacting intermediate (III-1-1-1) with sulfur monochloride, and then reacting under the action of aluminum trichloride:

[0051] wherein Z1 is S or O; Z2 is CH or N, X1 is S or Se; R2 is selected from Specifically:

[0052] The preparation method of the compound corresponding to formula (I) is as follows: X is O, R3 is H, and R2 is absent;

[0053] Or the preparation method is as follows: X is N, R3 is H;

[0054] The preparation method of the compound corresponding to formula (I) is as follows: X is S, R3 is H, and R2 is absent;

[0055] The preparation method of the compound corresponding to formula (I) is as follows: X is Se, R3 is H, and R2 is absent;

[0056] The preparation method of the compound corresponding to formula (II) is as follows: R3 is H;

[0057] Or the preparation method of the compound corresponding to formula (II) is as follows:

[0058] Or the preparation method of the compound corresponding to formula (III) is as follows:

[0059] The pyridinone derivative according to any one of the above is used for preparing an influenza drug.

[0060] The pyridinone derivative according to any one of the above is used for preparing an influenza drug.

[0061] The positive progress effect of the present application is that:

[0062] The hydroxypyridinone compound according to the present application can effectively inhibit the replication of influenza virus in vivo and in vitro. In particular, the compound according to the present application is less affected by the influenza PA / I38T mutant strain, and the resistance index (RI=IC 50 [I38T] / IC 50 [WT]) exhibited in the in vitro polymerase inhibition experiment is significantly lower than that of the positive baloxavir. The hydroxypyridinone compound according to the present application also has the application in preparing an anti-baloxavir-resistant virus drug and an anti-avian influenza H5N1 drug, has small cytotoxicity and high selectivity. The hydroxypyridinone compound with a novel structure disclosed in the present application has a good application prospect in the clinical treatment of baloxavir-resistant virus strains and avian influenza. BRIEF DESCRIPTION OF DRAWINGS

[0063] Fig. 1 is a general structural formula of the protected compound according to the present application;

[0064] Fig. 2 is a protection activity curve of compound 12-12-A in avian influenza H5N1 infected MDCK cells;

[0065] Fig. 3 is the antiviral activity research of compound 17-12-A and baloxavir BXM in the model of baloxavir-resistant A / WSN / 33 influenza strain infected BALB / c mice: (A) the body weight change of the model mice after being given compound 17-12-A or baloxavir BXM by gavage; (B) the lung virus titer results of the model mice after being given compound 17-12-A or baloxavir BXM by gavage. DETAILED DESCRIPTION

[0066] The present application will be further described in detail below in combination with examples, but the embodiments of the present application are not limited thereto. According to the ordinary technical knowledge and conventional means in the art, various substitutions and modifications can be made without departing from the technical idea of the present application, and all the substitutions and modifications shall be included in the scope of the present application.

[0067] Example 1. Synthesis of compounds 1-6-A and 1-6-B

[0068] Step one: synthesis of compound 1-2

[0069] To a solution of compound 1-1 (5.0 g, 29.4 mmol) in DMF (10 mL) was added potassium carbonate (8.1 g, 58.8 mmol) under ice-bath, then tert-butyl nitrite (7.1 mL, 60 mmol) was added dropwise under stirring, after the addition was completed, the reaction was returned to room temperature and warmed to 60 °C, and stirred for 12 h. After the reaction was completed, the reaction was added to water (100 mL), and extracted with ethyl acetate (3 x 100 mL), the organic phase was combined, washed with water (1 x 100 mL), saturated brine (1 x 100 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated to give compound 1-2 as a crude product. Finally, compound 1-2 was obtained as a pure product (5.6 g, yield 92%) by recrystallization from petroleum ether / ethyl acetate. 1 H NMR (500 MHz, CDC13) δ 7.72 (d, J = 8.3 Hz, 1H), 7.07 (d, J = 1.8 Hz, 1H), 7.04 (dd, J = 8.4, 1.8 Hz, 1H), 4.80 (d, J = 2.4 Hz, 2H), 2.61 (s, 3H), 2.59 (t, J = 2.4 Hz, 1H). HRMS (ESI): m / z calcd for C 11 H 10 ClO2 + [M+H] + : 209.0364, found: 209.0360.

[0070] Step two: synthesis of compound 1-3

[0071] In a sealed tube, compound 1-2 (4.16 g, 20 mmol) was dissolved in solution DMSO (5 mL), then tert-butyl nitrite (7.1 mL, 60 mmol) was added dropwise under stirring, after the addition was completed, the reaction was warmed to 80 °C and stirred for 4 h. After the reaction was completed, the reaction was added to water (50 mL), and extracted with ethyl acetate (3 x 50 mL), the organic phase was combined, washed with water (1 x 100 mL), saturated brine (1 x 100 mL), dried over anhydrous sodium sulfate, filtered, rotary evaporated, and the obtained crude product was purified by silica gel column chromatography to give compound 1-3 as a pure product (2.35 g, 50%). 1 H NMR (500 MHz, DMSO-d6) δ 9.21 (s, 1H), 8.13 (d, J = 8.7 Hz, 1H), 7.37 (dd, J = 8.7, 2.1 Hz, 1H), 7.30 (d, J = 2.1 Hz, 1H), 5.28 (s, 2H). HRMS (ESI): m / z calcd for C 11 H7ClNO3 + [M+H]+ :236.0109, found: 236.0101

[0072] Step three: synthesis of compound 1-4

[0073] Compound 1-3 (2.35 g, 10 mmol) was added to 15 mL of methanol under ice bath, then sodium borohydride (0.76 g, 20 mmol) was added slowly under stirring, the reaction was continued to stir at room temperature for 2 h. After the reaction was completed, 5 mL of water was added to quench the reaction, then most of the methanol was removed by reduced pressure. The water phase was extracted with dichloromethane (2 x 50 mL), the combined organic phase was washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated to give 1-4 as a crude product. Finally, compound 1-4 was obtained as a pure product (2.18 g, yield 92%) by recrystallization with ethyl acetate. 1 H NMR (500 MHz, DMSO-d6) δ 8.74 (d, J = 1.2 Hz, 1H), 7.52 (dd, J = 8.1, 0.7 Hz, 1H), 7.26 - 7.09 (m, 2H), 6.44 (d, J = 5.4 Hz, 1H), 5.93 (d, J = 5.3 Hz, 1H), 5.21 - 5.11 (m, 2H). HRMS (ESI): m / z calcd for C 11 H7ClNO2 + [M-OH] + :220.0160, found: 220.0162

[0074] Step four: synthesis of compound 1-5

[0075] Compound 1-4 (0.28 g, 1.2 mmol) was added to a solution of 50% (w / w)-T4P / EA (1 mL, 1.2 mmol) (n-butylphosphinic acid (50% in ethyl acetate)) and the mixture was stirred at room temperature. Then (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H-[1,4]diazepino[3,4-c]pyrido[2,1-f][1,2,4]triazine-6,8-dione (0.20 g, 0.6 mmol) and 2 mL of ethyl acetate were added and the reaction was heated to 110 °C and stirred for 5 hours. After the reaction was completed, the reaction was quenched by adding to 15 mL of saturated sodium bicarbonate solution and extracted with ethyl acetate (2 x 50 mL). The organic phases were combined and washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude was purified by column chromatography to give 58 mg and 52 mg (total yield 17%) of pure compound 1-5-A and 1-5-B, respectively. The resulting compounds were tested by liquid chromatography column and compound 1-5-A had a retention time of 7.10 min and compound 1-5-B had a retention time of 7.14 min. Compound 1-5-A and compound 1-5-B are two non-enantiomeric isomers of the above formula (1-5) for *C, and so on.

[0076] Compound 1-5-A: 1 H NMR (500 MHz, CDC13) δ 8.28 (s, 1H), 7.67 - 7.64 (m, 2H), 7.39 (dd, J = 8.0, 6.4 Hz, 2H), 7.37 - 7.28 (m, 3H), 7.26 (s, 1H), 6.98 (d, J = 7.6 Hz, 1H), 6.13 (d, J = 7.8 Hz, 1H), 5.58 (s, 1H), 5.53 - 5.46 (m, 2H), 5.38 (d, J = 10.6 Hz, 1H), 4.87 (dd, J = 14.8, 1.4 Hz, 1H), 4.65 (dd, J = 13.6, 2.6 Hz, 1H), 4.50 (dd, J = 9.9, 3.0 Hz, 1H), 3.83 (dd, J = 10.8, 3.0 Hz, 1H), 3.75 (dd, J = 11.9, 3.3 Hz, 1H), 3.33 - 3.24 (m, 2H), 2.99 (ddd, J = 13.4, 11.8, 3.5 Hz, 1H). HRMS (ESI): m / z calcd for C 28 H 24 ClN4O6 + [M+H] + : 547.1379, found: 547.1371 Compound 1-5-B: 1H NMR (500 MHz, CDC13) δ 8.30 (s, 1H), 7.61 - 7.56 (m, 2H), 7.38 - 7.33 (m, 2H), 7.32 - 7.28 (m, 1H), 7.12 (d, J = 2.1 Hz, 1H), 6.96 (d, J = 7.7 Hz, 1H), 6.78 (dd, J = 8.1, 2.1 Hz, 1H), 6.44 (d, J = 8.2 Hz, 1H), 5.79 (d, J = 7.7 Hz, 1H), 5.59 (d, J = 11.0 Hz, 1H), 5.48 - 5.43 (m, 2H), 5.41 (d, J = 11.0 Hz, 1H), 4.87 (dd, J = 14.7, 1.3 Hz, 1H), 4.79 (dd, J = 9.9, 3.0 Hz, 1H), 4.70 (dd, J = 13.3, 2.3 Hz, 1H), 3.80 (ddd, J = 19.6, 11.2, 3.1 Hz, 2H), 3.34 - 3.18 (m, 3H). HRMS (ESI): m / z calcd for C 28 H 24 ClN4O6 + [M+H] + :547.1379, found:547.1380

[0077] Step five: synthesis of compound 1-6-A

[0078] Compound 1-5-A (55 mg, 0.1 mmol) was added to 1 mL of N,N-dimethylacetamide, lithium chloride (42 mg, 1 mmol) was added, and the reaction was stirred at 80 °C for 6 h. After the reaction was completed, the reaction system was placed in an ice bath, and 0.5 mL of acetone, 2 mL of 0.5 M aqueous HCl, and 1 mL of water were added, and stirring was continued for 1 h. The solid precipitated in the reaction was collected by suction filtration and dried to obtain crude compound 1-6-A. Further recrystallization with isopropyl ether / trichloromethane gave pure compound 1-6-A (36 mg, yield 79%). White solid, mp 212.4-213.3 °C. 1H NMR (500 MHz, DMSO-d6) δ 8.95 (s, 1H), 7.37 (d, J = 9.5 Hz, 1H), 7.25 (d, J = 2.2 Hz, 1H), 7.10 (d, J = 8.3 Hz, 1H), 7.05 (d, J = 7.6 Hz, 1H), 5.92 (s, 1H), 5.66 (d, J = 7.6 Hz, 1H), 5.49 (d, J = 14.8 Hz, 1H), 4.94 (d, J = 14.8 Hz, 1H), 4.85 (d, J = 9.8 Hz, 1H), 4.43 (d, J = 13.4 Hz, 1H), 3.77 (dd, J = 10.8, 3.1 Hz, 1H), 3.73 (dd, J = 11.7, 3.3 Hz, 1H), 3.62 - 3.55 (m, 1H), 3.40-3.42 (m, 1H), 3.16 - 3.10 (m, 1H). 13 C NMR (125 MHz, DMSO-d6) δ 160.93, 158.20, 157.12, 157.06, 138.20, 135.71, 133.67, 130.81, 125.24, 124.65, 123.29, 115.81, 110.91, 70.23, 68.07, 65.57, 64.45, 61.82, 45.38, 22.81. HRMS (ESI): m / z calcd for C 21 H 18 ClN4O6 + [M+H] + : 457.0910, found: 457.0912

[0079] Step six: synthesis of compound 1-6-B

[0080] Compound 1-5-B (55 mg, 0.1 mmol) was added to 1 mL of N,N-dimethylacetamide, and lithium chloride (42 mg, 1 mmol) was added. The reaction was stirred at 80 °C for 6 h. After the reaction was completed, the reaction system was placed in an ice bath, and 0.5 mL of acetone, 2 mL of 0.5 M aqueous HC1, and 1 mL of water were added, and stirring was continued for 1 h. The solid precipitated in the reaction was collected by suction filtration and dried to obtain crude compound 1-6-B. Further recrystallization with isopropyl ether / trichloromethane gave pure compound 1-6-B (30 mg, yield 66%). White solid, mp 204.1-205.0 °C. 1H NMR (500 MHz, DMSO-d6) δ 11.66 (s, 1H), 8.80 (s, 1H), 7.68 (d, J = 8.2 Hz, 1H), 7.38 (d, J = 2.1 Hz, 1H), 7.32 (dd, J = 8.2, 2.2 Hz, 1H), 7.18 (d, J = 7.6 Hz, 1H), 5.84 (s, 1H), 5.76 (d, J = 7.7 Hz, 1H), 5.55 (d, J = 14.9 Hz, 1H), 4.91 (d, J = 14.9 Hz, 1H), 4.53 - 4.45 (m, 1H), 4.42 (d, J = 13.4 Hz, 1H), 3.68 (td, J = 10.5, 3.2 Hz, 2H), 3.63 - 3.55 (m, 1H), 3.39 (s, 1H), 3.15 - 3.09 (m, 1H). 13 C NMR (125 MHz, DMSO-d6) δ 160.87, 158.39, 156.34, 155.62, 138.55, 135.71, 133.40, 126.79, 125.20, 124.02, 116.14, 111.10, 79.26, 79.00, 78.74, 70.27, 68.06, 65.54, 64.73, 61.97, 45.37, 22.79. HRMS (ESI): m / z calcd for C 21 H 18 ClN4O6 + [M+H] + : 457.0910, found: 457.0910

[0081] Example 2. Synthesis of compound 2-6-A and 2-6-B

[0082] Referring to the method of Example 1, except that 4'-chloro-2'-hydroxyacetophenone (1-1) was replaced by 4-fluoro-2-hydroxyacetophenone, two diastereoisomers of compound 2-6 corresponding to compound 2-6-A and compound 2-6-B were finally obtained. Compound 2-6-A: white solid, mp 226.9-227.3 °C. 1H NMR (500 MHz, DMSO-d6) δ 8.95 (s, 1H), 7.45 - 7.29 (m, 1H), 7.05 (d, J = 3.1 Hz, 1H), 7.03 - 7.01 (m, 1H), 6.86 (m, 1H), 5.89 (m, 1H), 5.64 (m, 1H), 5.51 (d, J = 15.0 Hz, 1H), 4.93 (d, J = 14.8 Hz, 1H), 4.84 (m, 1H), 4.44 (d, J = 13.4 Hz, 1H), 3.75 (ddd, J = 22.0, 11.3, 3.3 Hz, 3H), 3.63 - 3.55 (m, 1H), 3.15 (d, J = 12.5 Hz, 1H). 13 C NMR (125 MHz, DMSO-d6) δ 164.68, 162.71, 160.91 (d, J = 11.8 Hz), 158.84, 157.35, 157.06, 138.19, 133.91 (d, J = 10.58 Hz), 122.45, 115.77, 111.42 (d, J = 21.50 Hz), 110.87 (d, J = 22.68 Hz), 110.58, 110.40, 68.12, 67.31, 65.62, 64.28, 61.72, 45.37, 22.81. HRMS (ESI): m / z calcd for C 21 H 18 FN4O6 + [M+H] + : 441.1205, found: 441.1206 Compound 2-6-B: white solid, mp 232.5-233.6 °C. 1 H NMR (500 MHz, DMSO-d6) δ 11.67 (s, 1H), 8.81 (s, 1H), 7.71 (dd, J = 8.5, 6.6 Hz, 1H), 7.17 (dd, J = 10.8, 7.5 Hz, 2H), 7.11 (td, J = 8.4, 2.7 Hz, 1H), 5.84 (s, 1H), 5.77 (d, J = 7.6 Hz, 1H), 5.55 (d, J = 14.8 Hz, 1H), 4.91 (d, J = 14.9 Hz, 1H), 4.49 (dd, J = 9.9, 3.2 Hz, 1H), 4.42 (d, J = 13.3 Hz, 1H), 3.67 (dt, J = 10.9, 5.5 Hz, 2H), 3.61 - 3.56 (m, 1H), 3.40 (d, J = 2.8 Hz, 1H), 3.15 - 3.08 (m, 1H). 13C NMR (125 MHz, DMSO-d6) δ 171.31, 160.88, 159.11 (d, J = 11.8 Hz), 156.33, 155.85, 152.81, 138.57, 133.63 (d, J = 10.58 Hz), 116.13, 115.86, 112.13 (d, J = 21.50 Hz), 111.40 (d, J = 22.68 Hz), 111.11, 70.19, 68.06, 65.53, 64.58, 61.99, 45.39, 22.80. HRMS (ESI): m / z calcd for C 21 H 18 FN4O6 + [M+H] + : 441.1205, found: 441.1206

[0083] Example 3. Synthesis of compound 3-6-A and 3-6-B

[0084] Referring to the method of Example 1, except that 4'-chloro-2'-hydroxyacetophenone (1-1) was replaced by 2'-hydroxyacetophenone, two diastereoisomers corresponding to compound 3-6, compound 3-6-A and compound 3-6-B, were finally obtained.

[0085] Compound 3-6-A: white solid, mp 228.3-229.5 °C. 1 H NMR (500 MHz, DMSO-d6) δ 8.90 (s, 1H), 7.40 (t, J = 7.7 Hz, 1H), 7.31 (d, J = 7.5 Hz, 1H), 7.15 (d, J = 7.9 Hz, 1H), 7.08 (d, J = 7.7 Hz, 1H), 7.02 (t, J = 7.3 Hz, 1H), 5.84 (s, 1H), 5.65 (d, J = 7.6 Hz, 1H), 5.46 (d, J = 14.9 Hz, 1H), 4.93 (dd, J = 10.0, 3.2 Hz, 1H), 4.87 - 4.81 (m, 1H), 4.43 (dd, J = 13.4, 2.6 Hz, 1H), 3.75 (ddd, J = 17.9, 11.2, 3.2 Hz, 2H), 3.60 (d, J = 10.4 Hz, 1H), 3.42 - 3.39 (m, 1H), 3.16 (ddd, J = 15.0, 11.9, 3.5 Hz, 1H). 13C NMR (125 MHz, DMSO-d6) δ 170.88, 160.87, 157.50, 157.35, 156.72, 152.55, 138.25, 132.28, 126.47, 124.91, 123.12, 116.29, 116.08, 110.89, 73.13, 70.26, 68.10, 65.56, 64.49, 62.61, 45.42. HRMS (ESI): m / z calcd for C 21 H 19 N4O6 + [M+H] + : 423.1300, found: 423.1304

[0086] Compound 3-6-B: white solid, mp 215.1-216.1 °C. 1 H NMR (500 MHz, Methanol-d4) δ 8.61 (s, 1H), 7.43 (td, J = 7.7, 1.7 Hz, 1H), 7.36 (d, J = 7.5 Hz, 1H), 7.19 (d, J = 8.0 Hz, 1H), 7.16 (dd, J = 7.6, 1.7 Hz, 1H), 7.06 (t, J = 7.4 Hz, 1H), 5.93 (d, J = 7.5 Hz, 1H), 5.76 (s, 1H), 5.48 (d, J = 14.9 Hz, 1H), 5.13 (dd, J = 10.0, 3.1 Hz, 1H), 4.61 (dd, J = 13.5, 2.5 Hz, 1H), 3.85 (ddd, J = 22.6, 11.3, 3.3 Hz, 2H), 3.56 (t, J = 10.5 Hz, 1H), 3.51 - 3.46 (m, 1H), 3.46 - 3.33 (m, 1H), 3.32 (d, J = 2.0 Hz, 1H). 13 C NMR (125 MHz, Methanol-d4) δ 162.06, 159.39, 158.52, 157.32, 154.04, 140.64, 133.86, 133.13, 128.32, 126.82, 124.75, 118.26, 112.64, 91.46, 72.20, 69.94, 67.39, 66.38, 65.31, 49.51, 47.09. HRMS (ESI): m / z calcd for C 21 H 19 N4O6 + [M+H] + : 423.1300, found: 423.1298

[0087] Example 4.4-9-A and 4-9-B synthesis

[0088] Step one: synthesis of compound 4-2

[0089] To a solution of compound 4-1 (5.0 g, 37.0 mmol) in ethanol (15 mL) was added di-tert-butyl dicarbonate (9.3 mL, 40.7 mmol) at room temperature, after the addition was completed, the reaction was raised to 65 °C and stirred for 24 h. After the reaction was completed, the reaction was rotary evaporated to get compound 4-2 crude (7.4 g, yield 85%) which was used directly for the next step. 1 H NMR (500 MHz, CDC13) δ 10.93 (s, 1H), 8.46 (dd, J = 8.6, 1.2 Hz, 1H), 7.85 (dd, J = 8.0, 1.6 Hz, 1H), 7.54 - 7.46 (m, 1H), 7.02 (ddd, J = 8.2, 7.2, 1.2 Hz, 1H), 2.64 (s, 3H), 1.52 (s, 9H). HRMS (ESI): m / z calcd for C 13 H 17 NaNO3 + [M+Na] + :258.1101, found:258.1109

[0090] Step two: synthesis of compound 4-3

[0091] NaH (1.44 g, 60 mmol) was added to a solution of compound 4-2 (7.05 g, 30 mmol) in anhydrous THF 30 mL under N2protection at ice bath, then it was raised to room temperature and stirred for 1 h. 3-Bromopropyne (3.9 mL, 45 mmol) was added dropwise to the reaction, which was refluxed overnight. After the reaction was completed, 10 mL saturated NH4Cl solution was added to quench the reaction, then most of the THF was removed under reduced pressure. The aqueous phase was extracted with ethyl acetate (2 x 50 mL), the combined organic phase was washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated to get 4-3 crude, which was purified by silica gel column chromatography to get 4-3 pure (5.0 g, yield 61%). 1 H NMR (500 MHz, CDC13) δ 7.64 (s, 1H), 7.52 - 7.33 (m, 3H), 4.77 - 4.10 (m, 2H), 2.52 (s, 3H), 2.24 (s, 1H), 1.41 (d, J = 95.1 Hz, 9H). HRMS (ESI): m / z calcd for C 16 H 19 NaNO3+ [M+Na] + : 296.1258, found: 296.1257

[0092] Step three: synthesis of compound 4-4

[0093] In a sealed tube, compound 4-3 (4.1 g, 15 mmol) was dissolved in solution DMSO (5 mL), then tert-butyl nitrite (5.3 mL, 45 mmol) was added dropwise under stirring, after the dropwise addition was completed, the reaction liquid was raised to 80°C and stirred for 4 h. After the reaction was completed, the reaction liquid was added to water (50 mL) and extracted with ethyl acetate (3 x 50 mL), the combined organic phase was washed with saturated brine (1 x 100 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated, and the obtained crude product was purified by silica gel column chromatography to obtain compound 4-4 pure product (1.9 g, yield 42%). 1 H NMR (500 MHz, CDC13) δ 8.56 (s, 1H), 8.14 (s, 1H), 7.61 (t, J = 7.4 Hz, 1H), 7.50-7.28 (m, 2H), 5.70-3.89 (m, 2H), 1.31 (s, 9H). HRMS (ESI): m / z calcd for C 16 H 16 NaN2O4 + [M+Na] + : 323.1003, found: 323.1002

[0094] Step four: synthesis of compound 4-5

[0095] Compound 4-4 (1.5 g, 5 mmol) was dissolved in 10 mL of anhydrous DCM at room temperature, then trifluoroacetic acid 5 mL was added dropwise under stirring, after the dropwise addition was completed, the reaction liquid was stirred at room temperature for 6 h. After the reaction was completed, the reaction liquid was rotary evaporated, the residue was dissolved in 20 mL of DCM, and saturated sodium bicarbonate solution was added to adjust the pH value to 7. The aqueous phase was extracted with DCM (3 x 20 mL), the combined organic phase was washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated, and the obtained crude product was recrystallized with petroleum ether / dichloromethane to obtain compound 4-5 pure product (0.76 g, yield 76%). 1H NMR (500 MHz, DMSO-d6) δ 9.03 (s, 1H), 7.99 (dd, J = 8.3, 1.7 Hz, 1H), 7.35 (ddd, J = 8.4, 5.2, 1.7 Hz, 2H), 6.91 (dd, J = 8.5, 1.1 Hz, 1H), 6.78 (ddd, J = 8.2, 6.8, 1.1 Hz, 1H), 4.23 (d, J = 3.8 Hz, 2H). HRMS (ESI): m / z calcd for C 11 H9N2O2 + [M+H] + : 201.0659, found: 201.0653

[0096] Step five: synthesis of compound 4-6

[0097] Compound 4-5 (0.76 g, 3.8 mmol) and DIPEA (0.7 mL, 4 mmol) were dissolved in 5 mL of dry DCM at room temperature, then acetyl chloride (0.32 mL, 4.5 mmol) was added dropwise under stirring, after the dropwise addition was completed, the reaction was stirred at room temperature for 2 h. After the reaction was completed, the reaction was diluted with 20 mL of DCM, then washed with dilute hydrochloric acid (0.5 M, 1 x 20 mL), saturated sodium bicarbonate (1 x 20 mL), saturated sodium chloride solution (1 x 20 mL), respectively, the organic phase was dried over anhydrous sodium sulfate, filtered, rotary evaporated, the obtained crude was slurried with ether to obtain compound 4-6 (0.8 g, yield 87%). 1 H NMR (500 MHz, CDCl3) δ 8.65 - 8.53 (m, 1H), 8.16 (dd, J = 7.8, 1.7 Hz, 1H), 7.70 (dd, J = 7.7, 1.7 Hz, 1H), 7.59 (dd, J = 7.7, 1.2 Hz, 1H), 7.35 (dd, J = 7.9, 1.2 Hz, 1H), 5.86 (d, J = 15.7 Hz, 1H), 4.04 (dd, J = 15.7, 1.5 Hz, 1H), 1.86 (s, 3H). HRMS (ESI): m / z calcd for C 13 H 11 N2O3 + [M+H] + : 243.0795, found: 243.0794

[0098] Step six: synthesis of compound 4-7

[0099] To a solution of compound 4-6 (0.8 g, 3.3 mmol) in 10 mL of methanol was added under ice bath, then sodium borohydride (0.25 g, 6.6 mmol) was added slowly under stirring. The reaction was continued to stir at room temperature for 2 h. After the reaction was completed, 5 mL of water was added to quench the reaction, then most of the methanol was removed by reduced pressure. The aqueous phase was extracted with dichloromethane (2 x 50 mL), the combined organic phase was washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 4-7 as a crude product. Finally, compound 4-7 was recrystallized from ethyl acetate to give pure product (0.72 g, 90% yield). 1 H NMR (500 MHz, DMSO-d6) δ 8.72 (d, J = 1.1 Hz, 1H), 7.58 (dd, J = 7.5, 1.6 Hz, 1H), 7.51 - 7.39 (m, 3H), 6.17 (d, J = 141.5 Hz, 1H), 5.80 (dd, J = 16.4, 1.2 Hz, 1H), 5.72 (s, 1H), 3.81 (dd, J = 16.4, 1.5 Hz, 1H), 1.76 (s, 3H). HRMS (ESI): m / z calcd for C 13 H 13 N2O3 + [M+H] + :245.0921, found:245.0918

[0100] Step Seven: Synthesis of compound 4-8-A and 4-8-B

[0101] To a solution of compound 4-7 (0.29 g, 1.2 mmol) in 50% (w / w)-T4P / EA (1 mL, 1.2 mmol) was added under ice bath, then (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H- [1,4]diazepino[3,4-c]pyrido[2,1-f][1,2,4]triazine-6,8-dione (0.20 g, 0.6 mmol) and 2 mL of ethyl acetate were added, the mixture was heated to 110 °C and stirred for 5 h. After the reaction was completed, the reaction was added to 15 mL of saturated sodium bicarbonate solution to quench, then extracted with ethyl acetate (2 x 50 mL). The combined organic phase was washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by column chromatography to give pure compound 4-8-A and 4-8-B as 46 mg and 40 mg (total yield 13%) respectively. The obtained compounds were detected by liquid chromatography column, compound 4-8-A had a retention time of 7.60 min and compound 4-8-B had a retention time of 7.70 min. Compound 4-8-A and compound 4-8-B were two non-corresponding isomers corresponding to compound 4-8 as shown above.

[0102] Compound 4-8-A: 1 H NMR (500 MHz, DMSO-d6) δ 9.22 (s, 1H), 8.04 (dd, J = 7.8, 1.7 Hz, 1H), 7.80 (td, J = 7.6, 1.7 Hz, 1H), 7.70 (d, J = 7.6 Hz, 1H), 7.65 - 7.60 (m, 2H), 7.58 - 7.54 (m, 2H), 7.39 (m, 1H), 7.26 (m, 2H), 6.22 (d, J = 7.6 Hz, 1H), 5.84 (s, 1H), 5.61 (m, 1H), 5.07 (d, J = 2.4 Hz, 2H), 4.80 (ddd, J = 12.6, 9.9, 4.6 Hz, 1H), 4.18 - 4.10 (m, 2H), 4.03 (td, J = 11.3, 4.4 Hz, 2H), 3.43 (td, J = 12.0, 3.1 Hz, 1H), 3.13 (dd, J = 11.4, 10.0 Hz, 1H), 2.95 (ddd, J = 13.7, 12.1, 4.3 Hz, 1H), 1.77 (s, 3H). HRMS (ESI): m / z calcd for C 30 H 28 N5O6 + [M+H] + : 554.2035, found: 554.2038

[0103] Compound 4-8-B: 1 H NMR (500 MHz, DMSO-d6) δ 8.72 (d, J = 1.1 Hz, 1H), 7.70 (d, J = 7.6 Hz, 1H), 7.57 (td, J = 7.8, 1.4 Hz, 3H), 7.53 - 7.39 (m, 2H), 7.38 - 7.34 (m, 2H), 7.32 - 7.28 (m, 2H), 6.22 (d, J = 7.6 Hz, 1H), 5.80 (s, 1H), 5.68 (m, 1H), 5.08 (d, J = 2.3 Hz, 2H), 4.83 - 4.77 (m, 1H), 4.15 (dd, J = 13.8, 2.9 Hz, 1H), 4.02 (dt, J = 11.2, 5.4 Hz, 2H), 3.81 (dd, J = 16.4, 1.5 Hz, 1H), 3.46 - 3.41 (m, 1H), 3.13 (dd, J = 11.4, 9.9 Hz, 1H), 2.95 (ddd, J = 13.8, 12.2, 4.3 Hz, 1H), 1.76 (s, 3H). HRMS (ESI): m / z calcd for C 30 H 28N5O6 + [M+H] + :554.2035, found:554.2040

[0104] Step eight: synthesis of compound 4-9-A

[0105] Compound 4-8-A (46 mg, 0.08 mmol) was added to 1 mL of N,N-dimethylacetamide, lithium chloride (34 mg, 0.8 mmol) was added, and the reaction was stirred at 80 °C for 6 h. After the reaction was completed, the reaction system was placed in an ice bath, and 0.5 mL of acetone, 2 mL of 0.5 M aqueous HC1, and 1 mL of water were added, and stirring was continued for 1 h. The solid precipitated in the reaction was collected by suction filtration and dried to obtain crude compound 4-9-A. Further recrystallization with isopropyl ether / chloroform gave pure compound 4-9-A (31 mg, yield 83%). White solid, mp 236.0-237.0 °C. 1 H NMR (500 MHz, DMSO-d6) δ 9.21 (s, 1H), 8.04 (dd, J = 7.8, 1.6 Hz, 1H), 7.80 (td, J = 7.6, 1.6 Hz, 1H), 7.64 - 7.32 (m, 3H), 6.07 (d, J = 7.4 Hz, 1H), 5.75 (s, 2H), 4.88 (td, J = 13.2, 11.7, 4.1 Hz, 1H), 4.20 - 3.95 (m, 3H), 3.5-3.75 (m, 1H), 3.22 (t, J = 10.8 Hz, 2H), 3.02 (td, J = 13.1, 4.1 Hz, 1H), 1.77 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 182.20, 170.72, 168.72, 162.38, 158.63, 158.52, 151.61, 141.43, 137.80, 135.30, 134.69, 131.27, 129.65, 129.03, 118.76, 118.29, 111.82, 65.54, 65.49, 64.78, 64.13, 54.95, 22.16. HRMS (ESI): m / z calcd for C 23 H 22 N5O6 + [M+H] + :464.1565, found:464.1569

[0106] Step nine: synthesis of compound 4-9-B

[0107] Compound 4-8-B (40 mg, 0.07 mmol) was added to 1 mL of N,N-dimethylacetamide, and lithium chloride (30 mg, 0.7 mmol) was added, and the reaction was stirred at 80 °C for 6 h. After the reaction was completed, the reaction system was placed in an ice bath, and 0.5 ml of acetone, 2 mL of 0.5 M aqueous HCl, and 1 mL of water were added, and stirring was continued for 1 h. The solid precipitated in the reaction was collected by suction filtration and dried to obtain crude compound 4-9-B. Further recrystallization from isopropyl ether / chloroform gave pure compound 4-9-B (27 mg, yield 82%). White solid, mp 230.6-231.2 °C. 1 H NMR (500 MHz, DMSO-d6) δ 8.70 (d, J = 18.0 Hz, 1H), 7.57 (dd, J = 7.4, 1.5 Hz, 1H), 7.50 (td, J = 7.5, 1.6 Hz, 1H), 7.43 (dtd, J = 17.9, 7.5, 1.5 Hz, 2H), 7.39 - 7.20 (m, 1H), 6.01 (d, J = 15.5 Hz, 1H), 5.80 (d, J = 16.7 Hz, 1H), 5.74 (d, J = 19.2 Hz, 1H), 4.82 (s, 1H), 4.12 - 3.98 (m, 3H), 3.81 (d, J = 16.4 Hz, 1H), 3.47 - 3.42 (m, 1H), 3.21 (t, J = 10.7 Hz, 1H), 2.97 (t, J = 12.8 Hz, 1H), 1.77 (d, J = 8.4 Hz, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 169.56, 168.79, 162.25, 161.17, 160.56, 155.86, 141.46, 138.41, 130.21, 130.09, 129.86, 129.01, 128.82, 128.53, 125.09, 115.37, 66.65, 65.64, 65.50, 65.13, 64.72, 38.34, 22.46. HRMS (ESI): m / z calcd for C 23 H 22 N5O6 + [M+H] + :464.1565, found:464.1570

[0108] Example 5

[0109] Step one: synthesis of compounds 5-1~5-5 was the same as example 4

[0110] Step two: synthesis of compound 5-6

[0111] Compound 5-5 (1.0 g, 5 mmol) and DIPEA (1.1 mL, 6 mmol) were dissolved in 5 mL of dry DCM at room temperature, and then chloroacetyl chloride (0.48 mL, 6 mmol) was added dropwise under stirring. After the dropwise addition was completed, the reaction solution was stirred at room temperature for 2 h. After the reaction was completed, the reaction solution was diluted with 20 mL of DCM, and then washed with dilute hydrochloric acid (0.5 M, 1 x 20 mL), saturated sodium bicarbonate (1 x 20 mL), and saturated sodium chloride solution (1 x 20 mL), respectively. The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product was slurried with diethyl ether to obtain pure compound 5-6 (1.3 g, 93% yield). 1 H NMR (500 MHz, CDC13) δ 8.62 (d, J = 1.4 Hz, 1H), 8.20 (dd, J = 7.9, 1.7 Hz, 1H), 7.75 (td, J = 7.6, 1.7 Hz, 1H), 7.64 (td, J = 7.6, 1.2 Hz, 1H), 7.46 (dd, J = 7.8, 1.2 Hz, 1H), 5.83 (d, J = 15.7 Hz, 1H), 4.12 (dd, J = 15.7, 1.5 Hz, 1H), 3.87 (d, J = 12.8 Hz, 1H), 3.78 (d, J = 12.8 Hz, 1H). HRMS (ESI): m / z calcd for C 13 H 10 N2ClO3 + [M+H] + :277.0375, found:277.0368

[0112] Step three: synthesis of compound 5-7

[0113] Compound 5-6 (1.1 g, 4 mmol) was dissolved in 5 mL of acetonitrile at room temperature, and then KSCN (1.2 g, 12 mmol) was added under stirring. The reaction solution was stirred at 60°C for 4 h. After the reaction was completed, the solvent was concentrated, and the resulting product was dissolved in 20 mL of ethyl acetate. The organic phase was washed with water (1 x 50 mL), saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Finally, the crude product was slurried with diethyl ether to obtain pure compound 5-7 (1.0 g, 88% yield). 1H NMR (500 MHz, DMSO-d6) δ 9.23 (s, 1H), 8.11 (dd, J = 7.9, 1.7 Hz, 1H), 7.80 (td, J = 7.6, 1.7 Hz, 1H), 7.72 (dd, J = 8.0, 1.3 Hz, 1H), 7.66 (td, J = 7.6, 1.3 Hz, 1H), 5.60 (d, J = 15.7 Hz, 1H), 4.42 (d, J = 15.1 Hz, 1H), 4.24 (dd, J = 15.7, 1.5 Hz, 1H), 3.85 (d, J = 15.1 Hz, 1H). HRMS (ESI): m / z calcd for C 14 H 10 N3SO3 + [M+H] + : 300.0438, found: 300.0430

[0114] Step four: Synthesis of compounds 5-8 ~ 5-10-A and 5-10-B was the same as Example 4

[0115] Compound 5-10-A: white solid, mp 239.0-240.0 °C. 1 H NMR (500 MHz, DMSO-d6) δ 9.23 (s, 1H), 8.11 (dd, J = 7.8, 1.5 Hz, 1H), 7.80 (t, J = 7.5 Hz, 1H), 7.72 (d, J = 7.8 Hz, 1H), 7.65 - 7.61 (m, 1H), 7.33 (d, J = 12.5 Hz, 1H), 6.07 (d, J = 7.4 Hz, 1H), 5.88 - 5.48 (m, 2H), 4.89 (ddd, J = 13.5, 10.1, 4.0 Hz, 1H), 4.41 (d, J = 15.1 Hz, 1H), 4.24 (d, J = 15.4 Hz, 1H), 4.17 - 3.99 (m, 4H), 3.84 (d, J = 15.0 Hz, 1H), 3.46 (d, J = 2.9 Hz, 1H), 3.22 (s, 1H), 3.01 (dd, J = 13.2, 4.2 Hz, 1H). 13C NMR (125 MHz, DMSO-d6) δ 181.42, 169.65, 165.44, 162.25, 158.99, 158.78, 151.38, 139.79, 138.35, 135.75, 134.69, 132.11, 129.92, 129.56, 119.47, 117.98, 113.35, 112.47, 65.72, 65.68, 64.91, 64.56, 40.56, 37.92. HRMS (ESI): m / z calcd for C 24 H 21 N6O6S + [M+H] + : 521.1238, found: 521.1240

[0116] Compound 5-10-B: white solid, mp 245.1-246.0 °C. 1 H NMR (500 MHz, DMSO-d6) δ 8.69 (s, 1H), 7.66 - 7.60 (m, 2H), 7.50 (dd, J = 6.9, 2.1 Hz, 1H), 7.43 (ddd, J = 7.0, 4.8, 1.9 Hz, 2H), 7.41 - 7.32 (m, 1H), 6.12 (s, 1H), 6.07 (d, J = 7.4 Hz, 1H), 4.89 (td, J = 15.7, 14.5, 8.5 Hz, 2H), 4.48 (d, J = 16.8 Hz, 1H), 4.09 (dd, J = 14.1, 2.9 Hz, 1H), 4.03 (td, J = 11.7, 5.5 Hz, 2H), 3.47 (dd, J = 12.2, 2.9 Hz, 1H), 3.28 (s, 2H), 3.23 (t, J = 10.7 Hz, 1H), 3.02 (td, J = 13.1, 4.1 Hz, 1H). 13 C NMR (125 MHz, DMSO-d6) δ 170.62, 167.50, 162.35, 161.22, 155.61, 151.56, 141.62, 137.73, 136.71, 129.42, 128.98, 128.77, 126.45, 125.17, 118.22, 114.04, 111.73, 103.00, 65.74, 65.50, 65.44, 64.74, 42.84, 40.81. HRMS (ESI): m / z calcd for C 24 H 21 N6O6S + [M+H] +:521.1238, found: 521.1234

[0117] Example 6. Synthesis of 6-6-A and 6-6-B

[0118] Step one: Synthesis of compound 6-2

[0119] Compound 6-1 (5.9 g, 30 mmol) was dissolved in 20 mL of acetone at room temperature, dimethyl sulfide (3.3 mL, 45 mmol) was added, and the reaction was stirred at room temperature overnight. After the reaction was completed, the reaction was filtered to obtain a white solid, which was dried to obtain compound 6-2 crude product (7.0 g, yield 90%), which was directly used in the next step.

[0120] Step two: Synthesis of compound 6-3

[0121] Compound 6-2 (5.2 g, 20 mmol) was added to 10 mL of chloroform in a sealed tube, sodium acetate (1.64 g, 20 mmol), phenylacetylene (2.2 mL, 20 mmol), and tert-butyl nitrite (7.1 mL, 60 mmol) were added in sequence, the reaction was heated to 60°C and stirred for 6 h. After the reaction was completed, most of the solvent was removed by rotary evaporation, the residue was dissolved in 20 mL of DCM, the organic phases were combined and washed with water (1 x 50 mL), saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 6-3 pure product (3.3 g, yield 66%). 1 H NMR (500 MHz, CDC13) δ 8.38-8.33 (m, 2H), 7.88-7.83 (m, 2H), 7.70-7.65 (m, 1H), 7.57-7.49 (m, 5H), 7.06 (s, 1H). HRMS (ESI): m / z calcd for C 16 H 11 NaNO2 + [M+Na] + :272.0682, found: 272.0683

[0122] Step three: Synthesis of compound 6-4

[0123] To a solution of compound 6-3 (2.5 g, 10 mmol) in 10 mL of methanol was added sodium borohydride (0.76 g, 20 mmol) slowly under stirring at ice bath. The reaction was allowed to warm to room temperature and stirred for 2 h. After completion of the reaction, 5 mL of water was added to quench the reaction and most of the methanol was removed under reduced pressure. The aqueous phase was extracted with dichloromethane (2 x 50 mL) and the combined organic phase was washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give 6-4 as a crude product. The pure compound 6-4 was obtained by recrystallization from petroleum ether / ethyl acetate (2.2 g, 90% yield). 1 H NMR (500 MHz, CDC13) δ 7.75 - 7.69 (m, 2H), 7.52 - 7.48 (m, 2H), 7.45 - 7.37 (m, 5H), 7.36 - 7.31 (m, 1H), 6.42 (s, 1H), 6.03 (d, J = 2.1 Hz, 1H), 3.02 (d, J = 3.3 Hz, 1H). HRMS (ESI): m / z calcd for C 16 H 12 NO + [M-OH] + : 234.0914, found: 234.0918

[0124] Step four: synthesis of compound 6-5-A and 6-5-B

[0125] To a solution of compound 6-4 (0.3 g, 1.2 mmol) in 50% (w / w)-T4P / EA (1 mL, 1.2 mmol) was added (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H- [1,4]diazepino[3,4-c]pyrido[2,1-f][1,2,4]triazine-6,8-dione (0.20 g, 0.6 mmol) and 2 mL of ethyl acetate in a sealed tube and the mixture was stirred at 110 °C. After 5 h, the reaction was quenched by adding to 15 mL of saturated sodium bicarbonate solution and extracted with ethyl acetate (2 x 50 mL). The combined organic phase was washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product. The pure compound 6-5-A and 6-5-B were obtained by column chromatography (30 mg and 24 mg, total yield 8% respectively). The compounds were detected by liquid chromatography column, compound 6-5-A retention time 7.0 min and compound 6-5-B retention time 7.1 min.

[0126] Compound 6-5-A: 1H NMR (500 MHz, Methanol-d4) δ 7.81 - 7.77 (m, 2H), 7.72 (d, J = 7.6 Hz, 1H), 7.51 (ddd, J = 7.7, 4.1, 1.9 Hz, 3H), 7.49 - 7.44 (m, 4H), 7.37 (dd, J = 8.4, 6.9 Hz, 2H), 7.31 (dq, J = 9.5, 7.3, 6.7 Hz, 4H), 6.70 (s, 1H), 6.46 (d, J = 7.5 Hz, 1H), 5.91 (s, 1H), 5.16 (d, J = 2.1 Hz, 2H), 4.64 (dd, J = 9.9, 4.7 Hz, 1H), 4.19 (dd, J = 14.0, 3.0 Hz, 1H), 4.05 (td, J = 12.1, 4.5 Hz, 2H), 3.52 (td, J = 11.9, 3.1 Hz, 1H), 3.17 (dd, J = 11.5, 9.9 Hz, 1H), 2.98 (ddd, J = 14.0, 12.2, 4.4 Hz, 1H). HRMS (ESI): m / z calcd for C 33 H 29 N4O5 + [M+H] + :561.2133, found: 561.2128

[0127] Compound 6-5-B: 1 H NMR (500 MHz, DMSO-d6) δ 8.23 - 8.17 (m, 2H), 8.03 - 7.98 (m, 2H), 7.79 - 7.75 (m, 1H), 7.70 (d, J = 7.6 Hz, 1H), 7.65 - 7.58 (m, 4H), 7.57 - 7.54 (m, 3H), 7.36 (dd, J = 8.1, 6.5 Hz, 2H), 7.32 - 7.28 (m, 2H), 6.22 (d, J = 7.6 Hz, 1H), 5.75 (s, 1H), 5.08 (d, J = 2.4 Hz, 2H), 4.82 - 4.77 (m, 1H), 4.15 (dd, J = 13.8, 2.8 Hz, 1H), 4.03 (dt, J = 11.3, 5.8 Hz, 2H), 3.47 - 3.41 (m, 1H), 3.13 (dd, J = 11.4, 10.0 Hz, 1H), 2.99 - 2.92 (m, 1H). HRMS (ESI): m / z calcd for C 33 H 29 N4O5 + [M+H] + :561.2133, found: 561.2130

[0128] Step five: synthesis of compound 6-6-A

[0129] Compound 6-5-A (30 mg, 0.053 mmol) was added to 1 mL of N,N- dimethylacetamide, lithium chloride (26 mg, 0.6 mmol) was added, and the reaction was stirred at 80 °C for 6 h. After the reaction was completed, the reaction system was placed in an ice bath, and 0.5 mL of acetone, 2 mL of 0.5 M aqueous HC1, and 1 mL of water were added, and stirring was continued for 1 h. The solid precipitated in the reaction was collected by suction filtration and dried to obtain a crude product. Further recrystallization with isopropyl ether / chloroform gave pure compound 6-6 (17 mg, yield 68%). White solid, mp 198.0-199.0 °C 1 H NMR (500 MHz, DMSO-d6) δ 7.87 - 7.84 (m, 2H), 7.62 (d, J = 7.5 Hz, 1H), 7.51 - 7.46 (m, 5H), 7.36 (q, J = 6.9, 6.2 Hz, 3H), 7.28 (t, J = 7.3 Hz, 1H), 6.98 (s, 1H), 6.07 (d, J = 7.4 Hz, 1H), 5.85 (s, 1H), 4.93 - 4.84 (m, 1H), 4.09 (dd, J = 14.1, 3.0 Hz, 1H), 4.03 (td, J = 11.7, 5.5 Hz, 2H), 3.46 (d, J = 2.9 Hz, 1H), 3.23 (s, 1H), 3.05 - 2.99 (m, 1H). 13 C NMR (125 MHz, DMSO-d6) δ 169.40, 168.56, 162.68, 162.45, 151.96, 142.57, 138.35, 131.00, 129.80, 128.92, 128.15, 127.16, 126.60, 126.00, 118.85, 112.51, 98.96, 67.95, 65.88, 65.83, 65.11. HRMS (ESI): m / z calcd for C 26 H 23 N4O5 + [M+H] + : 471.1663, found: 471.1666

[0130] Step six: synthesis of compound 6-6-B

[0131] Compound 6-5-B (24 mg, 0.043 mmol) was added to 1 mL of N,N- dimethylacetamide, lithium chloride (26 mg, 0.6 mmol) was added, and the reaction was stirred at 80 °C for 6 h. After the reaction was completed, the reaction system was placed in an ice bath, and 0.5 mL of acetone, 2 mL of 0.5 M aqueous HCL, and 1 mL of water were added, and stirring was continued for 1 h. The solid precipitated in the reaction was collected by suction filtration and dried to obtain a crude product. Further recrystallization from isopropyl ether / chloroform gave pure compound 6-6-B (13 mg, yield 64 %). White solid, mp 202.5-203.1 °C. 1 H NMR (500 MHz, DMSO-d6) δ 8.20 (d, J = 7.6 Hz, 2H), 8.02 (dd, J = 7.3, 2.1 Hz, 1H), 7.78 (t, J = 7.4 Hz, 1H), 7.73 (dd, J = 7.6, 1.6 Hz, 1H), 7.66 - 7.61 (m, 4H), 7.60 - 7.56 (m, 1H), 7.50 (s, 1H), 7.35 (d, J = 12.3 Hz, 1H), 6.07 (d, J = 7.4 Hz, 1H), 5.75 (s, 1H), 4.92 - 4.85 (m, 1H), 4.08 (td, J = 13.3, 12.7, 4.9 Hz, 2H), 4.03 - 4.00 (m, 1H), 3.45 (d, J = 2.9 Hz, 1H), 3.24 (d, J = 10.7 Hz, 1H), 3.04 - 2.98 (m, 1H). 13 C NMR (125 MHz, DMSO-d6) δ 185.40, 170.62, 167.14, 162.34, 161.89, 151.56, 151.08, 137.72, 135.28, 134.49, 132.49, 131.12, 131.01, 130.25, 130.08, 128.89, 128.07, 124.84, 118.21, 111.72, 104.92, 99.76, 65.50, 65.44, 64.74, 54.91. HRMS (ESI): m / z calcd for C 26 H 23 N4O5 + [M+H] + : 471.1663, found: 471.1658

[0132] Example 7. Synthesis of 7-6-A and 7-6-B

[0133] Referring to the method of Example 6, except that 2-bromoacetophenone (6-1) was replaced with a-bromo-4-chloroacetophenone, the total yield was 9 %.

[0134] Compound 7-6-A: white solid, mp 186.2-187.2 °C. 1 H NMR (500 MHz, DMSO-d6) δ 8.23 (m, 2H), 8.00 (dd, J = 7.6, 1.8 Hz, 3H), 7.70 (d, J = 8.6 Hz, 3H), 7.58 (m, 2H), 7.35 (d, J = 12.4 Hz, 1H), 6.07 (d, J = 7.4 Hz, 1H), 5.75 (s, 1H), 4.93 - 4.82 (m, 1H), 4.11 - 4.01 (m, 3H), 3.48 (m, 1H), 3.24 (d, J = 10.7 Hz, 1H), 3.04 - 2.99 (m, 1H). 13 C NMR (125 MHz, DMSO-d6) δ 185.69, 170.46, 169.44, 164.53, 162.54, 162.32, 162.27, 151.52, 137.79, 135.39, 134.47, 130.28, 128.89, 128.63, 128.56, 122.90, 118.38, 116.69, 116.51, 111.82, 100.96, 65.52, 65.46, 64.75, 54.93, 48.63. HRMS (ESI): m / z calcd for C 26 H 22 ClN4O5 + [M+H] + : 505.1274, found: 505.1272

[0135] Compound 7-6-B: white solid, mp 191.4-192.0 °C. 1 H NMR (500 MHz, DMSO-d6) δ 7.86 - 7.84 (m, 2H), 7.62 (d, J = 7.5 Hz, 1H), 7.53 - 7.47 (m, 5H), 7.42 (d, J = 1.9 Hz, 1H), 7.35 (d, J = 12.5 Hz, 1H), 6.98 (s, 1H), 6.07 (d, J = 7.4 Hz, 1H), 5.87 (s, 1H), 4.93 - 4.83 (m, 1H), 4.09 (dd, J = 14.0, 3.0 Hz, 1H), 4.03 (td, J = 11.7, 5.6 Hz, 2H), 3.48 - 3.43 (m, 1H), 3.24 (d, J = 10.7 Hz, 1H), 3.02 (td, J = 13.1, 4.2 Hz, 1H). 13C NMR (125 MHz, DMSO-d6) δ 184.59, 170.83, 170.55, 162.43, 162.18, 151.66, 139.56, 137.88, 134.13, 132.23, 131.27, 131.22, 129.53, 129.13, 128.87, 126.14, 126.05, 118.37, 111.93, 101.05, 65.59, 65.54, 64.83, 64.18, 63.73, 54.98. HRMS (ESI): m / z calcd for C 26 H 22 ClN4O5 + [M+H] + :505.1274, found:505.1278

[0136] Example 8. Synthesis of compound 8-8

[0137] Step one: Synthesis of compound 8-2

[0138] Compound 8-1 (3.5 g, 24.5 mmol) was dissolved in 10 mL of dry DCM and 100 μL of dry DMF, oxalyl chloride (4.2 mL, 49 mmol) was added dropwise slowly, the reaction was stirred at room temperature overnight. After the reaction was completed, the reaction was rotary evaporated to get compound 8-2 crude, which was used directly for the next step.

[0139] Step two: Synthesis of compound 8-3

[0140] Compound 8-2 was dissolved in 20 mL of dry DCM, hydroxylamine hydrochloride (3.0 g, 30 mmol) was added, DIPEA (8.7 mL, 50 mmol) was added dropwise slowly with stirring, the reaction was stirred at room temperature overnight. After the reaction was completed, 30 mL of 1 M HC1 was added to quench the reaction. The aqueous phase was extracted with dichloromethane (2 x 50 mL), the combined organic phase was washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated to get 8-3 crude. Finally, compound 8-3 pure (3.72 g, yield 83%) was obtained by recrystallization from petroleum ether / ethyl acetate.

[0141] 1 H NMR (500 MHz, CDC13) δ 8.77 (s, 1H), 3.67 (s, 3H), 3.34 (s, 3H), 2.77 (s, 3H).

[0142] HRMS (ESI): m / z calcd for C7H 11 N2O2S +[M+H] + : 187.0536, found: 187.0530

[0143] Step three: synthesis of compound 8-4

[0144] A magnesium ribbon with surface oxide film removed was cut into small pieces (1.46 g, 60 mmol) and added to 10 mL of anhydrous THF, a few iodine crystals were added under stirring, the solution quickly turned to brown-black. To the reaction system, 2-bromo-5-fluorotoluene (1.26 mL, 10 mmol) in THF 10 mL was added, and the temperature was raised to reflux until the reaction was successfully initiated, at which time the solution color disappeared, close to colorless transparent. After removing the heat, the reaction was stirred at room temperature, and the remaining 2-bromo-5-fluorotoluene (5 mL, 40 mmol) in THF 10 mL was continuously added through a dropping funnel, and the system was kept slightly refluxed to continue the reaction for about 2 h to obtain a 1 M / L THF solution of 4-fluoro-2-methylphenyl magnesium bromide.

[0145] Compound 8-3 (2.8 g, 15 mmol) was dissolved in 10 mL of anhydrous THF, and the above prepared 4-fluoro-2-methylphenyl magnesium bromide THF solution 20 mL was added, and the reaction was stirred at room temperature for 2 h. After the reaction was completed, 0.5 M HCl was added to the system until the system showed weak acidity, and the reaction was continuously stirred for 30 min to quench the reaction. The aqueous phase was extracted with ethyl acetate (2 x 50 mL), and the combined organic phase was washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated to obtain the crude product, which was purified by silica gel column chromatography to obtain pure compound 8-4 (1.65 g, yield 47%). 1 H NMR (500 MHz, CDC13) δ 8.84 (s, 1H), 7.40 (dd, J = 8.5, 5.8 Hz, 1H), 7.02 - 6.93 (m, 2H), 2.54 (s, 3H), 2.39 (s, 3H). HRMS (ESI): m / z calcd for C 12 H 11 FNOS + [M+H] + : 236.0540, found: 236.0536

[0146] Step four: synthesis of compound 8-5

[0147] Compound 8-4 (0.8 g, 3.4 mmol) was added to 10 mL of methanol under ice bath, then sodium borohydride (0.26 g, 6.8 mmol) was added slowly under stirring, the reaction was continued to stir at room temperature for 2 h. After the reaction was completed, 5 mL of water was added to quench the reaction, then most of the methanol was removed by reduced pressure. The water phase was extracted with dichloromethane (2 x 50 mL), the combined organic phase was washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated to give the crude 8-5. Finally, the compound 8-5 was obtained by recrystallization from petroleum ether / ethyl acetate (0.8 g, yield 99%). 1 H NMR (500 MHz, CDC13) δ 8.54 (s, 1H), 7.55 (dd, J = 8.6, 5.9 Hz, 1H), 6.94 (td, J = 8.4, 2.7 Hz, 1H), 6.85 (dd, J = 9.6, 2.7 Hz, 1H), 6.16 (d, J = 3.4 Hz, 1H), 3.16 (d, J = 3.6 Hz, 1H), 2.40 (s, 3H), 2.22 (s, 3H). HRMS (ESI): m / z calcd for C 12 H 13 FNOS + [M+H] + :238.0697, found:238.0694

[0148] Step five: synthesis of compound 8-6

[0149] Compound 8-5 (0.5 g, 2.1 mmol) was dissolved in 5 mL of anhydrous DCM, and phosphorus tribromide (0.32 mL, 3.4 mmol) was added slowly, the reaction was stirred at room temperature for 6 h. After the reaction was completed, the reaction system was quenched by adding cold saturated sodium bicarbonate solution, the water phase was extracted with dichloromethane (2 x 50 mL), the combined organic phase was washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated to give the crude 8-6, which was used directly in the next step. HRMS (ESI): m / z calcd for C 12 H 11 FNS + [M-Br] + :220.0591, found:220.0590

[0150] Step six: synthesis of compound 8-7

[0151] Compound 8-6 (0.3 g, 1 mmol) was added to 5 mL of dry acetonitrile, followed by (R)-7-benzyloxy-3,4,12,12A-tetrahydro-lH-[l,4]diazepino[3,4-C]pyrido[2,l-F][l,2,4]triazine-6,8-dione (0.27 g, 0.83 mmol) and CsF (0.32 g, 2.1 mmol), the reaction was heated to 70 °C and stirred for 12 h. After the reaction was completed, the reaction was quenched by adding to 25 mL of saturated sodium bicarbonate solution and extracted with ethyl acetate (2 x 50 mL). The combined organic phase was washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude was purified by column chromatography to give a mixture of diastereomers of compound 8-7 (23 mg, 5% yield). HRMS (ESI): m / z calcd for C 29 H 28 N4O4FS + [M+H] + : 547.1810, found: 547.1804

[0152] Step seven: synthesis of compound 8-8

[0153] Compound 8-7 (20 mg, 0.036 mmol) was added to 1 mL of N,N-dimethylacetamide, lithium chloride (26 mg, 0.6 mmol) was added, and the reaction was stirred at 80 °C for 6 h. After the reaction was completed, the reaction was placed in an ice bath, and 0.5 mL of acetone, 2 mL of 0.5 M aqueous HC1, and 1 mL of water were added, and stirring was continued for 1 h. The solid precipitated in the reaction was collected by suction filtration and dried to give the crude product. Further recrystallization from isopropyl ether / chloroform gave a mixture of diastereomers of compound 8-8 (11 mg, 65% yield).

[0154] White solid, mp 217.3-218.1 °C. A mixture of diastereomers (1.7: 1) was obtained in 65% yield. 1H NMR (500 MHz, DMSO-d6) δ 8.83 (s, 1.7H), 8.11 (s, 1H), 7.70 (d, J = 7.6 Hz, 1H), 7.62 (dd, J = 6.9, 3.1 Hz, 2H), 7.56 (dd, J = 7.0, 1.6 Hz, 1.7H), 7.52 (dd, J = 8.6, 6.2 Hz, 1.7H), 7.39 - 7.35 (m, 2H), 7.35 - 7.29 (m, 3.4H), 7.06 (s, 1H), 7.05 - 7.02 (m, 1.7H), 7.00 (d, J = 3.2 Hz, 1H), 6.99 - 6.96 (m, 1.7H), 6.23 (d, J = 7.6 Hz, 1H), 6.07 (d, J = 7.5 Hz, 1.7H), 6.06 (s, 1.7H), 5.91 (s, 1H), 5.07 (d, J = 2.8 Hz, 1.7H), 4.95 - 4.82 (m, 1.7H), 4.82 - 4.71 (m, 1H), 4.18 - 4.06 (m, 3H), 4.03 (dtd, J = 11.5, 6.7, 6.2, 2.7 Hz, 5.1H), 3.46 (s, 1H), 3.44 (d, J = 2.6 Hz, 1.7H), 3.25 - 3.22 (m, 1.7H), 3.15 (d, J = 1.4 Hz, 1H), 3.05 - 2.99 (m, 1.7H), 2.97 (s, 1H), 2.35 (s, 5.1H), 2.17 (s, 5.1H), 2.13 (s, 3H), 2.02 (s, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 174.03, 170.73, 162.36, 162.11 (d, J = 1.76 Hz), 160.17 (d, J = 2.65 Hz), 151.72, 150.37, 148.35, 146.46, 139.63, 138.83 (d, J = 2.77 Hz), 137.73, 137.46, 137.40, 136.18, 135.9 (d, J = 2.4 Hz), 128.13, 128.07, 127.68, 127.33, 118.24, 116.68, 116.53, 116.51 (d, J = 2.77 Hz), 116.37, 115.29, 112.45, 112.32, 112.28, 112.16, 111.72, 72.76, 65.80, 65.52, 65.47, 65.41, 64.76, 64.17, 64.03, 62.33, 18.55, 18.33, 15.12, 11.16. HRMS (ESI): m / z calcd for C 22 H 22N4O4FS + [M+H] + : 457.1341, found: 457.1336

[0155] Example 9. Synthesis of 9-8

[0156] The procedure of Reference Example 8 was followed except that 4-methylthiazole-5- carboxylic acid was replaced by 4-methyl-5-carboxylic acidyl-1,3-oxazole to give finally a mixture of diastereomers of formula 9-8 in a ratio of 2:1 (yield 66%).

[0157] Compound 9-8: white solid, mp 229.6-230.3 °C. 1 H NMR (500 MHz, DMSO-d6) δ 8.56 (s, 2H), 8.10 (s, 1H), 7.70 (d, J = 7.6 Hz, 1H), 7.62 (dd, J = 6.9, 2.3 Hz, 2H), 7.61 - 7.53 (m, 4H), 7.38 - 7.36 (m, 2H), 7.35 - 7.32 (m, 2H), 7.32 - 7.28 (m, 1H), 7.24 (dd, J = 10.2, 2.6 Hz, 2H), 7.16 (td, J = 8.6, 2.6 Hz, 2H), 7.05 (td, J = 8.7, 2.8 Hz, 1H), 6.98 (dd, J = 10.0, 2.8 Hz, 1H), 6.23 (d, J = 7.6 Hz, 1H), 6.07 (d, J = 7.5 Hz, 2H), 5.91 (s, 1H), 5.75 (s, 2H), 5.11 - 5.04 (m, 2H), 5.01 - 4.82 (m, 2H), 4.81 - 4.68 (m, 1H), 4.20 - 4.07 (m, 3H), 4.03 (tq, J = 11.3, 3.6, 2.7 Hz, 6H), 3.49 - 3.46 (m, 2H), 3.46 (d, J = 2.8 Hz, 1H), 3.25 - 3.21 (m, 2H), 3.17 - 3.13 (m, 1H), 3.06 - 2.99 (m, 2H), 2.96 - 2.90 (m, 1H), 2.31 (s, 6H), 2.28 (s, 6H), 2.12 (s, 3H), 2.02 (s, 3H). 13C NMR (125 MHz, DMSO-d6) δ 184.10, 174.03, 170.75, 164.16, 162.36, 160.20, 155.95, 153.78, 150.38, 148.75, 146.47, 145.82, 144.37, 139.96 (d, J = 8.8 Hz), 139.63 (d, J = 8.8 Hz), 137.67 (d, J = 7.3 Hz), 137.11 (d, J = 7.3 Hz), 135.93 (d, J = 3.2 Hz), 133.90 (d, J = 2.8 Hz), 131.35, 131.00, 128.07, 127.99, 118.24, 117.86, 117.69, 116.53, 116.37, 115.30, 112.75, 112.57, 112.32, 112.15, 111.72, 72.77, 65.80, 65.47, 64.76, 64.03, 62.33, 19.12, 18.33, 13.55, 11.16. HRMS (ESI): m / z calcd for C 22 H 22 N4O5F + [M+H] + : 441.1569, found: 441.1572

[0158] Example 10. Synthesis of 10-7

[0159] Following the procedure of Example 8, except replacing 4-methylthiazole-5- carboxylic acid with 4-methylthiophene-5-carboxylic acid, the final product was a mixture of diastereomers as shown in 10-7 (18 mg, 72% yield) with a ratio of 1.6:1 for the two diastereomers.

[0160] White solid, mp 209.2-210.0 °C. 1H NMR (500 MHz, DMSO-d6) δ 7.91 (d, J = 5.0 Hz, 1.6 H), 7.70 (d, J = 7.6 Hz, 1 H), 7.62 (d, J = 7.6 Hz, 1.6 H), 7.57 (d, J = 1.6 Hz, 1 H), 7.55 (s, 1 H), 7.49 (dd, J = 8.6, 6.2 Hz, 1 H), 7.44 (dd, J = 8.5, 5.9 Hz, 1.6 H), 7.38 - 7.35 (m, 2 H), 7.34 - 7.29 (m, 3.2 H), 7.26 (d, J = 5.1 Hz, 1 H), 7.21 (dd, J = 10.2, 2.7 Hz, 1.6 H), 7.15 - 7.11 (m, 3.2 H), 7.01 (td, J = 8.7, 2.9 Hz, 1 H), 6.97 (dd, J = 10.1, 2.8 Hz, 1 H), 6.82 (d, J = 5.0 Hz, 1 H), 6.23 (d, J = 7.6 Hz, 1 H), 6.07 (d, J = 7.5 Hz, 1.6 H), 6.00 (s, 1.6 H), 5.75 (s, 1 H), 5.07 (d, J = 3.0 Hz, 2 H), 4.88 (ddd, J = 12.5, 10.2, 4.3 Hz, 1.6 H), 4.83 - 4.77 (m, 1 H), 4.17 - 4.07 (m, 3 H), 4.02 (dq, J = 11.3, 3.4, 2.7 Hz, 4.8 H), 3.44 (s, 2.6 H), 3.23 (s, 1 H), 3.16 - 3.12 (m, 1.6 H), 3.02 (dd, J = 4.2, 1.5 Hz, 1 H), 2.95 (ddd, J = 13.7, 12.1, 4.3 Hz, 1.6 H), 2.28 (s, 4.8 H), 2.25 (s, 4.8 H), 2.17 (s, 3 H), 2.14 (s, 3 H). 13C NMR (125 MHz, DMSO-d6) δ 189.57, 174.03, 170.71, 163.65, 162.36, 161.68, 155.94, 151.61, 148.74, 145.74, 141.87, 139.62, 138.95, 138.47, 137.74, 136.85, 136.37, 133.84, 133.07, 131.34, 129.97, 129.56, 129.49, 127.67, 123.54, 118.24, 117.52, 117.35, 116.45, 116.28, 115.29, 112.58, 112.41, 112.15, 111.98, 111.76, 72.77, 65.80, 65.47, 65.03, 64.76, 64.03, 18.96, 18.55, 16.11, 13.55. HRMS (ESI): m / z calcd for C 23 H 23 FN3O4S + [M+H] + :456.1388, found:456.1386

[0161] Example 11. Synthesis of 11-8-A and 11-8-B

[0162] Step one: Synthesis of compound 11-2

[0163] To a solution of compound 11-1 (10.0 g, 76.9 mmol) in DCM (100 mL) was added pyridine (6.7 g, 84.6 mmol), then acetyl chloride (6.0 mL, 84.6 mmol) was added dropwise under stirring, after the addition was completed, the reaction was stirred at room temperature for 12 h. After the reaction was completed, the reaction was poured into 2M HC1 (100 mL), and extracted with DCM (3x100 mL), the organic phase was washed with water (1x100 mL) successively, saturated brine (1x100 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated to give the crude product. Purified by silica gel column chromatography to give compound 11-2 (12 g, yield 96%). 1 H NMR (500 MHz, CDCl3) δ 7.11–7.03 (m, 2H), 6.95–6.88 (m, 1H), 2.35 (s, 3H). HRMS (ESI): m / z calcd for C8H6F2O2Na + [M+Na] + :195.0229, found:195.0228

[0164] Step two: synthesis of compound 11-3

[0165] Compound 11-2 (12.0 g, 70 mmol) was dissolved in solution DCE (50 mL) at 0 °C, then anhydrous aluminum trichloride (9.3 g, 70 mmol) was added portionwise under stirring, after the dropwise addition was completed, the reaction was raised to reflux and stirred for 12 h. After the reaction was completed, 2M HC1 (100 mL) was added to quench the reaction, and extracted with DCM (3 x 100 mL), the organic phase was combined, washed with water (1 x 100 mL) and saturated brine (1 x 100 mL) successively, dried over anhydrous sodium sulfate, filtered, and rotary evaporated, the obtained crude product was purified by silica gel column chromatography to give compound 11-3 (9.7 g, 81%). 1 H NMR (500 MHz, CDCl3) δ 12.57 (d, J = 1.3 Hz, 1H), 7.52 (ddd, J = 9.2, 5.5, 2.3 Hz, 1H), 6.72 (td, J = 9.3, 6.6 Hz, 1H), 2.62 (s, 3H). HRMS (ESI): m / z calcd for C8H6F2O2Na + [M+Na] + : 195.0229, found: 195.0231

[0166] Step three: synthesis of compound 11-4~11-8 Refer to the method of Example 1, except that 4'-chloro-2'-hydroxyacetophenone (1-1) was replaced by compound 11-3, and finally two non-corresponding isomers of compound 11-8 corresponding to compound 11-8-A and compound 11-8-B were obtained.

[0167] Compound 11-8-A: white solid. 1 H NMR (500 MHz, DMSO-d6) δ 11.68 (s, 1H), 8.98 (s, 1H), 7.20 (t, J = 7.6 Hz, 1H), 7.15 - 7.08 (m, 1H), 7.06 (d, J = 7.7 Hz, 1H), 5.99 (s, 1H), 5.69 (d, J = 7.7 Hz, 1H), 5.58 (d, J = 14.6 Hz, 1H), 5.07 (d, J = 14.7 Hz, 1H), 4.81 (dd, J = 9.9, 3.1 Hz, 1H), 4.44 (dd, J = 13.5, 2.5 Hz, 1H), 3.78 (dd, J = 10.8, 3.1 Hz, 1H), 3.73 (dd, J = 11.6, 3.3 Hz, 1H), 3.59 (t, J = 10.4 Hz, 1H), 3.44 - 3.40 (m, 1H), 3.16 - 3.10 (m, 1H).13 C NMR (125 MHz, DMSO-d6) δ 177.04, 160.96, 157.33, 156.81, 151.65, 138.22, 127.02, 126.90, 124.06, 123.93, 119.90, 115.53, 112.57, 112.44, 110.96, 110.85, 70.20, 68.03, 65.54, 64.55, 61.81, 45.37. HRMS (ESI): m / z calcd for C 21 H 17 F2N4O6 + [M+H] + : 459.1111, found: 459.1111 Compound 11-8-B: white solid. HRMS (ESI): m / z calcd for C 21 H 17 F2N4O6 + [M+H] + : 459.1111, found: 459.1108

[0168] Example 12. Synthesis of 12-12-A and 12-12-B

[0169] Step one: Synthesis of compound 12-5

[0170] Compound 12-4 (4 g, 25.8 mmol) was dissolved in 15 mL of chlorobenzene, and the solution was heated to 100 °C with stirring, then BPO (0.62 g, 2.58 mmol) and dibromohydantoin (8.87 g, 31.0 mmol) were added to the reaction solution, and the solution was heated to reflux and reacted overnight. After the reaction was completed, chlorobenzene was removed by concentration under reduced pressure, and the residue was dissolved in DCM, and insoluble matter was removed by filtration. The filtrate was rotary evaporated to obtain a crude product, which was purified by silica gel column chromatography to obtain pure compound 12-5 (3 g, yield 51%). 1 H NMR (500 MHz, CDCl3) δ 8.59 (s, 1H), 4.56 (s, 2H), 4.48 (q, J = 7.1 Hz, 2H), 1.45 (t, J = 7.1 Hz, 3H). HRMS (ESI): m / z calcd for C7H9NBrO3 + [M+H] + : 233.9761, found: 233.9759

[0171] Step two: Synthesis of compound 12-6

[0172] Compound 12-5 (2 g, 8.6 mmol) and triphenylphosphine (2.7 g, 10.3 mmol) were added to 15 mL of toluene under nitrogen atmosphere and the reaction was refluxed for 4 h. After the reaction was completed, the filtrate was removed by filtration and the filter cake was washed with toluene to obtain crude compound 12-6 which was used directly in the next step without further purification. HRMS (ESI): m / z calcd for C 25 H 23 NPO3 + [M+H] + :416.1411, found:416.1412

[0173] Step three: synthesis of compound 12-7

[0174] Compound 12-6 crude (1.5 g, 3.0 mmol) was dissolved in 10 mL of anhydrous acetonitrile at 0 °C, then 2,3-difluorobenzaldehyde (0.85 g, 6.0 mmol) and DBU (0.55 g, 3.6 mmol) were added successively under stirring, the reaction was warmed to reflux and reacted overnight. After the reaction was completed, the solvent was removed by concentration under reduced pressure, and the compound 12-7 pure (0.8 g, yield 94%) was directly purified by silica gel column chromatography. 1 H NMR (500 MHz, CDC13) δ 8.09 (d, J = 0.9 Hz, 1H), 7.11 (dtd, J = 10.0, 7.7, 2.0 Hz, 1H), 7.06 - 6.93 (m, 2H), 6.87 (d, J = 12.0 Hz, 1H), 6.72 (d, J = 12.0 Hz, 1H), 4.45 (q, J = 7.1 Hz, 2H), 1.43 (t, J = 7.2 Hz, 3H). HRMS (ESI): m / z calcd for C 14 H 12 NF2O3 + [M+H] + :280.0780, found:280.0773

[0175] Step four: synthesis of compound 12-8

[0176] Compound 12-7 (0.8 g, 2.9 mmol) was dissolved in ethanol / water (5 mL / 1 mL), LiOH H2O (0.24 g, 5.8 mmol) was added under stirring, and the reaction was allowed to proceed at room temperature for 2 h. After the reaction was completed, ethanol was removed by concentration under reduced pressure, 2 M HC1 was added dropwise to the residue under stirring until the pH of the system reached about 6. Filtration was performed under suction, and the filtrate was discarded. The filter cake was washed with water, and dried to obtain 0.65 g of crude compound 12-8, which was used directly in the next step without further purification. HRMS (ESI): m / z calcd for C 12 H8NF2O3 + [M+H] + :252.0467, found: 252.0461

[0177] Step five: synthesis of compound 12-9

[0178] Compound 12-8 (0.65 g, 2.6 mmol) was dissolved in 5 mL of anhydrous DCM, and sulfurous chloride (0.38 mL, 5.2 mmol) was added dropwise under stirring, followed by the addition of a catalytic amount of DMF. The reaction was allowed to proceed at reflux for 4 h. After the reaction was completed, 10 mL of carbon tetrachloride and anhydrous aluminum trichloride (0.78 g, 5.2 mmol) were added to the system after the system was allowed to return to room temperature, and the reaction was allowed to proceed at reflux overnight. After the reaction was completed, 10 mL of 1 M HC1 was added dropwise to the system to quench the reaction, and the aqueous phase was extracted with DCM (1 x 50 mL). The combined organic phases were washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated to obtain crude 12-9. Finally, compound 12-9 was purified by silica gel column chromatography (0.55 g, yield 92%). 1 H NMR (500 MHz, CDC13) δ 8.97 (s, 1H), 8.35 (ddd, J = 9.2, 5.5, 2.0 Hz, 1H), 7.37 (td, J = 8.9, 7.5 Hz, 1H), 7.28 (s, 1H), 7.07 (d, J = 12.0 Hz, 1H). HRMS (ESI): m / z calcd for C 12 H6NF2O2 + [M+H] + :234.0362, found: 234.0363

[0179] Step six: synthesis of compound 12-10

[0180] Compound 12-9 (0.55 g, 2.36 mmol) was added to 10 mL of ethanol under ice bath, then sodium borohydride (0.09 g, 2.36 mmol) was added slowly under stirring, the reaction was continued to stir at room temperature for 2 h. After the reaction was completed, 5 mL of water was added to quench the reaction, then most of the ethanol was removed by reduced pressure. The aqueous phase was extracted with dichloromethane (2 x 50 mL), the combined organic phase was washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product. Finally, the pure product of compound 12-10 was obtained by recrystallization with ethyl acetate (0.48 g, yield 87%). 1 H NMR (500 MHz, DMSO-d6) δ 9.13 (s, 1H), 7.48 (dq, J = 17.4, 8.5 Hz, 2H), 7.04 (d, J = 11.8 Hz, 1H), 6.97 (d, J = 11.8 Hz, 1H), 6.17 (s, 1H), 5.84 (s, 1H). HRMS (ESI): m / z calcd for C 12 H6NF2O + [M-OH] + : 218.0412, found: 218.0410

[0181] Step seven: synthesis of compound 12-11

[0182] Compound 12-10 (0.4 g, 1.7 mmol) was added to a solution of 50% (w / w)-T4P / EA (2 mL, 2.4 mmol) (n-butylphosphinic acid (50% in ethyl acetate)) and the mixture was stirred at room temperature. Then (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H-[1,4]diazepino[3,4-c]pyrido[2,1-f][1,2,4]triazin-6,8-dione (0.26 g, 0.8 mmol) and 2 mL of ethyl acetate were added, the reaction was heated to 110 °C and stirred for 5 hours. After the reaction was completed, the reaction was added to 15 mL of saturated sodium bicarbonate solution for quenching, extracted with ethyl acetate (2 x 50 mL). The combined organic phase was washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, concentrated to give the crude product, which was purified by column chromatography (dichloromethane:methanol = 100:1→50:1) to give the pure products of compound 12-11-A and 12-11-B as 103 mg and 73 mg (total yield 42%) respectively. The obtained compounds were detected by liquid chromatography column, the retention time of compound 12-11-A was 7.10 min and the retention time of compound 12-11-B was 7.13 min. Compound 12-11-A and compound 12-11-B are two non-corresponding isomers of the above formula (12-11) about *C.

[0183] Compound 12-11-A: 1 H NMR (500 MHz, DMSO-d6) δ 9.40 (s, 1H), 7.58 - 7.54 (m, 2H), 7.41 - 7.36 (m, 2H), 7.35 - 7.30 (m, 1H), 7.27 - 7.20 (m, 1H), 7.17 - 7.10 (m, 2H), 7.07 (dd, J = 11.8, 1.5 Hz, 1H), 6.78 (d, J = 7.7 Hz, 1H), 6.19 (s, 1H), 5.72 (d, J = 7.7 Hz, 1H), 5.25 (d, J = 11.0 Hz, 1H), 5.08 (d, J = 11.0 Hz, 1H), 4.45 (dd, J = 13.5, 2.5 Hz, 1H), 4.22 (dd, J = 9.9, 3.1 Hz, 1H), 3.66 (dd, J = 11.6, 3.3 Hz, 1H), 3.53 (dd, J = 10.7, 3.1 Hz, 1H), 3.29 (d, J = 10.3 Hz, 1H), 3.22 (td, J = 11.7, 2.7 Hz, 1H), 2.97 (ddd, J = 13.4, 11.7, 3.5 Hz, 1H). HRMS (ESI): m / z calcd for C 29 H 23 N4F2O5 + [M+H] + : 545.1632, found: 545.1631

[0184] Compound 12-11-A: 1H NMR (500 MHz, DMSO-d6) δ 9.26 (s, 1H), 7.66 (dd, J = 8.8, 5.1 Hz, 1H), 7.63 - 7.57 (m, 1H), 7.55 - 7.51 (m, 2H), 7.38 - 7.34 (m, 2H), 7.32 - 7.29 (m, 1H), 7.11 (d, J = 11.9 Hz, 1H), 7.04 (dd, J = 11.8, 1.5 Hz, 1H), 6.74 (d, J = 7.6 Hz, 1H), 6.13 (s, 1H), 5.84 (d, J = 7.7 Hz, 1H), 5.17 (d, J = 10.8 Hz, 1H), 5.10 (d, J = 10.8 Hz, 1H), 4.39 (dd, J = 13.4, 2.5 Hz, 1H), 4.09 (dd, J = 9.8, 3.1 Hz, 1H), 3.62 (dd, J = 11.6, 3.3 Hz, 1H), 3.55 (dd, J = 10.6, 3.1 Hz, 1H), 3.22 - 3.12 (m, 2H), 2.86 (ddd, J = 13.5, 11.7, 3.5 Hz, 1H). HRMS (ESI): m / z calcd for C 29 H 23 N4F2O5 + [M+H] + : 545.1632, found: 545.1628

[0185] Step eight: synthesis of compound 12-12-A

[0186] Compound 12-11-A (93 mg, 0.17 mmol) was added to 1 mL of N,N- dimethylacetamide, lithium chloride (42 mg, 1 mmol) was added, and the reaction was stirred at 80 °C for 6 h. After the reaction was completed, the reaction system was placed in an ice bath, and 0.5 mL of acetone, 2 mL of 0.5 M aqueous HC1, and 1 mL of water were added, and stirring was continued for 1 h. The solid precipitated in the reaction was collected by suction filtration and dried to obtain crude compound 12-12-A. Further recrystallization with isopropyl ether / trichloromethane gave pure compound 12-12-A (70 mg, yield 90%).

[0187] White solid. 1H NMR (500 MHz, DMSO-d6) δ 11.70 (s, 1H), 9.41 (s, 1H), 7.38 - 7.32 (m, 1H), 7.28 (dd, J = 9.0, 5.1 Hz, 1H), 7.17 - 7.06 (m, 2H), 6.69 (d, J = 7.7 Hz, 1H), 6.21 (s, 1H), 5.56 (d, J = 7.6 Hz, 1H), 4.43 (dd, J = 13.3, 2.4 Hz, 1H), 4.24 (dd, J = 9.4, 3.7 Hz, 1H), 3.68 (dd, J = 11.6, 3.4 Hz, 1H), 3.56 - 3.48 (m, 2H), 3.40 - 3.37 (m, 1H), 3.05 (ddd, J = 13.3, 11.7, 3.5 Hz, 1H). 13 C NMR (125 MHz, DMSO-d6) δ 171.60, 161.54, 160.99, 158.79, 153.38, 138.92, 129.12, 128.93, 126.50, 120.71, 120.53, 120.44, 117.69, 117.56, 116.13, 115.54, 110.93, 70.12, 68.23, 65.96, 63.56, 45.82. HRMS (ESI): m / z calcd for C 22 H 17 N4F2O5 + [M+H] + : 455.1162, found: 455.1160

[0188] Step six: synthesis of compound 12-12-B

[0189] Compound 12-11-B (63 mg, 0.12 mmol) was added to 1 mL of N,N- dimethylacetamide, lithium chloride (42 mg, 1 mmol) was added, and the reaction was stirred at 80 °C for 6 h. After the reaction was completed, the reaction system was placed in an ice bath, and 0.5 mL of acetone, 2 mL of 0.5 M aqueous HC1, and 1 mL of water were added, and stirring was continued for 1 h. The solid precipitated in the reaction was collected by suction filtration and dried to obtain crude compound 12-12-B. Further recrystallization with isopropyl ether / trichloromethane gave pure compound 12-12-B (50 mg, yield 94%). White solid. 1H NMR (500 MHz, DMSO-d6) δ 11.68 (s, 1H), 9.27 (s, 1H), 7.70 (dd, J = 8.9, 5.1 Hz, 1H), 7.60 (q, J = 8.8 Hz, 1H), 7.12 (d, J = 11.8 Hz, 1H), 7.06 (dd, J = 11.7, 1.5 Hz, 1H), 6.66 (d, J = 7.6 Hz, 1H), 6.16 (s, 1H), 5.69 (d, J = 7.6 Hz, 1H), 4.45 - 4.33 (m, 1H), 4.14 (dd, J = 9.7, 3.2 Hz, 1H), 3.64 (dd, J = 11.7, 3.2 Hz, 1H), 3.55 (dd, J = 10.7, 3.2 Hz, 1H), 3.47 (m, 1H), 3.35 (m, 1H), 2.99 - 2.90 (m, 1H). 13 C NMR (125 MHz, DMSO-d6) δ 171.33, 160.97, 159.79, 157.64, 153.10, 138.21, 128.76, 128.53, 125.68, 125.61, 120.30, 119.88, 117.89, 117.75, 115.53, 115.46, 110.70, 69.18, 67.73, 65.43, 63.43, 45.35. HRMS (ESI): m / z calcd for C 22 H 17 N4F2O5 + [M+H] + : 455.1162, found: 455.1166.

[0190] Example 13. Synthesis of 13-16-A and 13-16-B

[0191] Step one: Synthesis of compound 13-10

[0192] The synthesis of compound 13-10 was performed according to the procedure described in Example 12.

[0193] Step two: Synthesis of compound 13-11

[0194] Compound 13-5 (1,2-bis(2,3-difluorophenyl)disulfide) (0.6 g, 2 mmol) was dissolved in acetonitrile / water (8 mL / 2 mL) at 0 °C under nitrogen atmosphere, and zinc powder (0.52 g, 8 mmol) was added. Iron trichloride (0.65 g, 4 mmol) was added in batches under stirring, the reaction was restored to room temperature and continued to react for 3 h. Then compound 13-10 (0.9 g, 4 mmol) was added and reacted overnight. After the reaction was completed, the insoluble was filtered off by diatomite, and the filter cake was washed with EA, the filtrate was collected, and washed with water (1x50 mL), saturated brine (1x50 mL) in turn. After drying over anhydrous sodium sulfate and concentrating under reduced pressure, the crude product was obtained, which was then purified by silica gel column chromatography (petroleum ether: ethyl acetate = 50:1) to obtain pure compound 13-11 (0.7 g, yield 91%).

[0195] 1 H NMR (500 MHz, CDC13) δ 8.28 (s, 1H), 7.13-6.97 (m, 3H), 4.46 (q, J = 7.1 Hz, 2H), 4.19 (d, J = 0.9 Hz, 2H), 1.43 (t, J = 7.2 Hz, 3H).

[0196] HRMS (ESI): m / z calcd for C 13 H 12 SF2NO3 + [M+H] + :300.0501, found:300.0495

[0197] Step three: synthesis of compound 13-12

[0198] Compound 13-11 (0.7 g, 2.3 mmol) was dissolved in ethanol / water (5 mL / 1 mL), and LiOH H2O (0.2 g, 4.6 mmol) was added under stirring at room temperature, and reacted for 2 h. After the reaction was completed, ethanol was removed by concentrating under reduced pressure, and 2M HCl was added dropwise to the residue under stirring until the pH of the system reached about 6. Filtration was performed, and the filtrate was discarded, and the filter cake was washed with water, and dried to obtain crude compound 13-12 0.56 g, which was directly used in the next step without further purification.

[0199] HRMS (ESI): m / z calcd for C 11 H8SF2NO3 + [M+H] + :272.0188, found:272.0184

[0200] Step four: synthesis of compound 13-13

[0201] Compound 13-12 (0.5 g, 1.8 mmol) was added to 10 g of polyphosphoric acid and stirred at 150 °C for 24 h. After the reaction was completed, the reaction solution was slowly dropped into a cold saturated sodium bicarbonate solution, the aqueous phase was extracted with EA (2 x 100 mL), and the combined organic phase was washed with saturated brine (1 x 50 mL). After drying over anhydrous sodium sulfate, it was rotary evaporated to obtain a crude product. The pure compound 13-13 (0.2 g, yield 46%) was obtained after purification by silica gel column chromatography.

[0202] 1 H NMR (500 MHz, CDC13) δ 8.49 (s, 1H), 7.88 (ddd, J = 8.9, 5.3, 1.9 Hz, 1H), 7.30 - 7.27 (m, 1H), 3.95 (s, 2H).

[0203] HRMS (ESI): m / z calcd for C 11 H6SF2NO2 + [M+H] + : 254.0082, found: 254.0082

[0204] Step five: synthesis of compound 13-14

[0205] Compound 13-13 (0.2 g, 0.8 mmol) was added to 10 mL of ethanol under ice bath, then sodium borohydride (0.06 g, 1.6 mmol) was slowly added under stirring, and the reaction solution was continued to be stirred at room temperature for 2 h. After the reaction was completed, 5 mL of water was added to quench the reaction, and then most of the ethanol was removed by rotary evaporation under reduced pressure. The aqueous phase was extracted with dichloromethane (2 x 50 mL), and the combined organic phase was washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated to obtain a crude product. Finally, the pure compound 13-14 (0.18 g, yield 90%) was obtained by recrystallization with ethyl acetate.

[0206] 1 H NMR (500 MHz, DMSO-d6) δ 8.69 (s, 1H), 7.54 - 7.42 (m, 2H), 6.62 (d, J = 4.6 Hz, 1H), 6.33 (d, J = 4.6 Hz, 1H), 4.08 - 3.98 (m, 2H).

[0207] HRMS (ESI): m / z calcd for C 11 H6SF2NO + [M-OH] + : 238.0133, found: 238.0131

[0208] Step six: synthesis of compounds 13-15

[0209] Compound 13-14 (0.18 g, 0.7 mmol) was added to a solution of 50% (w / w)-T4P / EA (2 mL, 2.4 mmol) (n-butylphosphinic acid (50% in ethyl acetate)) followed by (R)-7-benzyloxy-3,4,12,12a-tetrahydro-lH-[l,4]diazepino[3,4-c]pyrido[2,l- f][l,2,4]triazine-6,8-dione (0.16 g, 0.5 mmol) and 2 mL of ethyl acetate, the reaction was heated to 110 °C and stirred for 5 hours. After the reaction was completed, the reaction was quenched by adding to 15 mL of saturated sodium bicarbonate solution and extracted with ethyl acetate (2 x 50 mL). The organic phases were combined and washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude was purified by column chromatography to give pure compound 13-15-A and 13-15-B as 20 mg and 14 mg (total yield 12%) respectively. The resulting compounds were detected by liquid chromatography column, compound 13-15-A had a retention time of 7.02 min and compound 13-15-B had a retention time of 7.04 min. Compound 13-15-A and compound 13-15-B are two non-enantiomeric isomers of the above formula (13-15) about *C.

[0210] Compound 13-15-A: HRMS (ESI): m / z calcd for C 28 H 22 N4F2O5S + [M+H] + : 565.1352, found: 565.1350

[0211] Compound 13-15-B: HRMS (ESI): m / z calcd for C 28 H 22 N4F2O5S + [M+H] + : 565.1352, found: 565.1349

[0212] Step eight: synthesis of compound 13-16-A

[0213] Compound 13-15-A (20 mg, 0.035 mmol) was added to 1 mL of N,N- dimethylacetamide, lithium chloride (15 mg, 0.35 mmol) was added, and the reaction was stirred at 80 °C for 6 h. After the reaction was completed, the reaction system was placed in an ice bath, and 0.5 mL of acetone, 2 mL of 0.5 M aqueous HC1, and 1 mL of water were added, and stirring was continued for 1 h. The solid precipitated in the reaction solution was collected by suction filtration and dried to obtain crude compound 13-16-A. Further recrystallization from isopropyl ether / chloroform gave pure compound 13-16-A (15 mg, yield 90%).

[0214] White solid. HRMS (ESI): m / z calcd for C 21 H 17 N4F2O5S + [M+H] + : 475.0883, found: 475.0880

[0215] Step nine: synthesis of compound 13-16-B

[0216] Compound 13-15-B (14 mg, 0.025 mmol) was added to 1 mL of N,N- dimethylacetamide, lithium chloride (11 mg, 0.25 mmol) was added, and the reaction was stirred at 80 °C for 6 h. After the reaction was completed, the reaction system was placed in an ice bath, and 0.5 mL of acetone, 2 mL of 0.5 M aqueous HC1, and 1 mL of water were added, and stirring was continued for 1 h. The solid precipitated in the reaction solution was collected by suction filtration and dried to obtain crude compound 13-16-B. Further recrystallization from isopropyl ether / chloroform gave pure compound 13-16-B (11 mg, yield 90%).

[0217] White solid. HRMS (ESI): m / z calcd for C 21 H 17 N4F2O5S + [M+H] + : 475.0883, found: 475.0878

[0218] Example 14. Synthesis of 14-15-A and 14-15-B

[0219] Referring to the method of Example 13, compound 13-5 (1,2-bis(2,3- difluorophenyl)disulfide) was replaced by compound 13-5 (1,2-bis(2,3- difluorophenyl)diselenide), and finally two diastereoisomers corresponding to compound 14-15, compound 14-15-A and compound 14-15-B (see the claims or the specification for structures, the same below), were obtained.

[0220] Compound 14-15-A: HRMS (ESI): m / z calcd for C 21 H 17 N4F2O5Se + [M+H] + : 523.0327, found: 523.0325

[0221] Compound 14-15-B: HRMS (ESI): m / z calcd for C 21 H 17 N4F2O5Se + [M+H] + : 523.0327, found: 523.0330

[0222] Example 15. Synthesis of 15-12-A and 15-12-B

[0223] Following the procedure of Example 12, 2,3-difluorobenzaldehyde was replaced by benzaldehyde, resulting in the final two diastereoisomers of compound 15-12, compound 15-12-A and compound 15-12-B.

[0224] Compound 15-12-A: HRMS (ESI): m / z calcd for C 22 H 19 N4O5[M+H] + : 419.1350, found: 419.1352

[0225] Compound 15-12-B: HRMS (ESI): m / z calcd for C 22 H 19 N4O5[M+H] + : 419.1350, found: 419.1353

[0226] Example 16. Synthesis of 16-12-A and 16-12-B

[0227] Following the procedure of Example 12, 2,3-difluorobenzaldehyde was replaced by 4-fluorobenzaldehyde, resulting in the final two diastereoisomers of compound 16-12, compound 16-12-A and compound 16-12-B.

[0228] Compound 16-12-A: HRMS (ESI): m / z calcd for C 22 H 18 FN4O5[M+H] +: 437.1256, found: 437.1257

[0229] Compound 16-12-B: HRMS (ESI): m / z calcd for C 22 H 18 FN4O5[M+H] + : 437.1256, found: 437.1257

[0230] Example 17 Inhibition effect of the compound disclosed in the present application on HEK-293T cell-expressed influenza virus polymerase

[0231] After trypsin digestion and counting, the HEK-293T cells were suspended in DMEM medium containing 10% fetal bovine serum, and then inoculated into a 96-well plate at 1.5 x 10 4 cells / well with a volume of 100 μL. The 96-well plate was incubated at 37°C / 5% CO2for 24 hours. After 24 hours, 20 μL of DMEM solution containing a certain concentration of the test compound was added, and 20 μL of DMEM solution was added to the control well, and the incubation was continued in the cell incubator for 2 hours. Then 100 μL / well of PBS solution was added to gently wash the cells, and after removing the PBS, 1 x CLB cell lysis solution was added to each well, and after shaking at room temperature for 20 minutes, the lysis solution in the plate was transferred to a 1.5 mL centrifuge tube, which was centrifuged at a speed of 12000 rpm for 2 minutes, and 20 μL of supernatant was placed in a 96-well white plate, and then Luciferase substrate and Renilla substrate were added in sequence, and the luminescence value was detected by an enzyme marker. The regression curve and regression equation were obtained by regression analysis, and the IC 50 value of the compound disclosed in the present application was calculated. In Table 1: PA / WT represents wild-type influenza PA; PA / I38T represents I38T mutant influenza PA.

[0232] Table 1. IC values and RI values of the hydroxypyridinone compounds and baloxavir disclosed in the present application 50

[0233] Example 1-16 and IC 50 ​The values of EC50 and RI are shown in Table 1. The results show that the hydroxypyridinone compounds of the present application can effectively inhibit the RNA polymerase of influenza A virus in HEK-293T cells, especially the compounds prepared in Examples 14 and 15, but the inhibitory activity of baloxavir on the PA / I38T resistant strain decreases significantly. The hydroxypyridinone compounds disclosed in the present application have a smaller drug resistance index RI than baloxavir, indicating that the derivatives of the present application are less affected by the PA / I38T mutant strain of influenza A, and are superior to baloxavir.

[0234] Antiviral effect of the compounds disclosed in the present application in MDCK cells infected with influenza A virus

[0235] MDCK cells were trypsinized and counted, then suspended in DMEM medium containing 10% fetal bovine serum, and then inoculated into 24-well plates at a concentration of 1.5 x 10 5 cells / well, with a medium volume of 500 μL per well. The 24-well plates were incubated at 37°C / 5% CO2 for 24 hours, and then the medium was removed and 500 μL of fresh medium was added. After adding a certain concentration of the test compound, the influenza virus was inoculated into the 24-well plates at a multiplicity of infection (MOI = virus number / cell number) of 0.1, and incubated in a cell culture incubator for 24 hours. The supernatant was collected and the viral TCID 50 values were determined. Regression curves and regression equations were obtained by regression analysis, and the EC 50 values of the compounds of the present application were calculated.

[0236] Table 2. EC 50 values of the hydroxypyridinone compounds of the present application and baloxavir

[0237] The EC 50 values of Examples 1, 2, 12 and baloxavir are shown in Table 2. The results show that the hydroxypyridinone compounds (1-6-A, 1-6-B, 2-6-A, 2-6-B, 12-12-A, 12-12-B) obtained in the present application can effectively inhibit influenza A virus in MDCK cells, and Examples 1, 2 and 12 have inhibitory activity comparable to baloxavir for the wild-type strain, and Example 12 has significantly better inhibitory activity than baloxavir for the I38T mutant strain.

[0238] Antiviral effect of the compounds disclosed in the present application in MDCK cells infected with avian influenza virus H5N1

[0239] MDCK cells were trypsinized and counted, then suspended in DMEM medium containing 10% fetal bovine serum, and then inoculated into 24-well plates at a concentration of 1.5 x 10 4MDCK cells were seeded in 48-well plates at 5 x 105cells / well in a volume of 200 μL per well. The 48-well plates were incubated at 37 °C / 5% CO2for 24 hours and the medium was removed and replaced with 200 μL of fresh medium. The compound to be tested was added at a certain concentration, and then the influenza virus was inoculated into the 48-well plates at a multiplicity of infection (MOI = virus number / cell number) of 0.01, and the plates were incubated in the cell incubator for 24 hours. The supernatant was collected and tested by the virus plaque assay to calculate the EC 50 value of the compound according to the present application.

[0240] As shown in Figure 2, the hydroxypyridinone compound 12-12-A can effectively inhibit the replication of avian influenza virus in MDCK cells, and the EC 50 value is 11.45 nM.

[0241] Example 20. Synthesis of 17-12-A

[0242] At room temperature, 12-12-A (0.45 g, 1 mmol), chloromethyl dimethyl carbonate (0.25 g, 2 mmol), potassium carbonate (0.27 g, 2 mmol) and potassium iodide (0.17 g, 1 mmol) were added to 5 mL of DMA, and the reaction was carried out at 60 °C for 4 h. After the reaction was completed as monitored by TLC, 50 mL of water was added to quench the reaction, and the aqueous phase was extracted with EA (50 mL x 2), and the combined organic phase was washed with saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, rotary evaporated, and purified by column chromatography (DCM:MeOH = 50:1) to obtain 17-12-A.

[0243] White solid, yield: 84%, m.p. 266.1-266.9 °C

[0244] 1 H NMR (500 MHz, CDC13) δ 8.59 (s, 1H), 7.33-7.27 (m, 1H), 7.25-7.22 (m, 1H), 7.07 (dd, J = 11.8, 1.7 Hz, 1H), 6.96 (d, J = 11.8 Hz, 1H), 6.53 (d, J = 7.8 Hz, 1H), 6.00 (d, J = 7.7 Hz, 1H), 5.92-5.85 (m, 3H), 4.55 (dd, J = 13.5, 2.5 Hz, 1H), 4.17 (dd, J = 9.8, 3.0 Hz, 1H), 3.82 (s, 3H), 3.70 (ddd, J = 10.8, 5.8, 3.2 Hz, 2H), 3.36 (td, J = 11.8, 2.7 Hz, 1H), 3.30 (t, J = 10.4 Hz, 1H), 2.88 (ddd, J = 13.5, 11.8, 3.5 Hz, 1H).

[0245] 13 C NMR (125 MHz, CDC13) δ 174.3, 157.9, 157.4, 155.2, 154.9, 151.5 (dd, J = 252.5, 13.6 Hz), 149.7, 149.2 (dd, J = 252.5, 13.6 Hz), 139.5, 128.3, 128.0 (d, J = 3.6 Hz), 127.6 (dd, J = 7.1, 5.4 Hz), 126.2 (d, J = 8.5 Hz), 121.2 (dd, J = 9.7, 2.9 Hz), 118.9 (d, J = 1.4 Hz), 118.0 (d, J = 17.4 Hz), 115.9, 115.3, 90.5, 69.6, 68.9, 66.5, 65.4, 55.0, 46.3.

[0246] HRMS (ESI): m / z calcd for C 25 H 21 N4F2O8 + [M+H] + : 543.1322, found: 543.1324.

[0247] Example 21. Antiviral effect of the compound 17-12-A of the present disclosure in a mouse model of influenza A virus infection

[0248] Using 6-8 week old female BALB / c mice, the mice were lightly anesthetized using isoflurane, and then inoculated intranasally with 50 μL of maraviroc-resistant A / WSN / 33 influenza strain (5 x 10 5 TCID 50 / mouse) dissolved in CMC-Na. 100 μL of the test compound or blank control (vehicle: 5% DMSO + 35% PEG400 + 5% Tween80 + 55% physiological saline) was given to the mice by gavage within 2h. 8 mice were set in each group for the control group and the treatment group, and the body weight change was monitored every day. The body weight change of each mouse was calculated as the percentage of the body weight on the day to the initial body weight. The mice were sacrificed on the 3rd day (72 hours after virus infection) and lung tissues were collected. The lung homogenate was subjected to freeze-thaw cycle and vortexed thoroughly to promote virus release, and the process was repeated three times. After freezing and thawing, the lung homogenate was centrifuged at 12000 rpm for 10 minutes at 4°C, and the supernatant was collected and added to MDCK cells to test the lung influenza virus TCID 50 .

[0249] As shown in Figure 3, the lung virus titers of the mice in the 17-12-A (20 mg / kg and 50 mg / kg) administration groups were significantly lower than those of the mice in the Vehicle group without administration, and the lung virus titers of the mice in the high and low dose 17-12-A administration groups were lower than those of the mice administered with marbooxavir, and the body weights of the mice in the administration groups were not significantly affected, indicating that 17-12-A can effectively inhibit the infection of marbooxavir-resistant A / WSN / 33 influenza virus.

Claims

1. A pyridone derivative, characterized in that, having the structure of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, wherein R1 is selected from hydrogen, halogen, C1-C6 alkyl, R1 is one or independently multiple; R2is selected from the absence, C1-C6alkyl, C1-C3alkylacyl or R3is selected from hydrogen or R4 is selected from hydrogen or C1-C6 alkyl; X is selected from N, O, S, Se; when X is O, S, Se, R2 is absent; A ring is selected from one of the following structures:

2. The pyridinone derivative according to claim 1, characterized in that, the corresponding C atom in formula (I), formula (II) or formula (III) is in the S configuration or in the R configuration or is a mixture comprising the S configuration and the R configuration: R3, R4 and X are defined as in claim 1; R1 is H, halogen, C1-C3 alkyl, R1 is one or independently two; R2is selected from the group consisting of absent, C1-C3alkyl, 3. The pyridinone derivative according to claim 1, characterized by R1is one, two or, independently of one another, three, each R1is each independently selected from the group consisting of hydrogen, F, CI, methyl, ethyl; R2is selected from the group consisting of absent, or R3is selected from the group consisting of hydrogen or R4is selected from the group consisting of hydrogen or methyl, ethyl; X is selected from the group consisting of N or O or S or Se; R2is absent when X is O, S, Se.

4. The pyridinone derivative according to claim 1 or 2, characterized in that, having the structure of Formula (I-a), (I-b), (I-c), (III-a) below: R1is H, halogen, C1-C3alkyl, R1is 1 or independently 2, R3is H or 5. The pyridinone derivative according to claim 1, wherein having a structure represented by any one of the following formulas:

6. An intermediate for preparing the pyridone derivative according to any one of claims 1 to 5, characterized in that, having a structure represented by any one of the following formulas: R1, R2, R4 and X are defined as in any one of claims 1-5.

7. The intermediate of claim 6, wherein, having a structure represented by any one of the following formulas:

8. A method for synthesizing the pyridinone derivative according to any one of claims 1 to 5, characterized by, comprising: The compound of formula (I-1) or formula (II-1) or formula (III-1) is reduced by sodium borohydride to obtain the intermediate of formula (I-2) or formula (II-2) or formula (III-2), and the intermediate (I-2) or the intermediate (II-2) or formula (III-2) is reacted with (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H-[1,4]azepino[3,4-c]pyrido[2,1-f][1,2,4]triazine-6,8-dione in the presence of n-butyl phosphoric anhydride to obtain the intermediate (I-3) or the intermediate (II-3) or the intermediate (III-3), and the intermediate (I-3) or (II-3) or the intermediate (III-3) is subjected to LiCl reduction to remove the benzyl group to obtain the derivative of pyridone of formula (I) or formula (II) or formula (III):

9. The method of synthesis of pyridinone derivatives according to claim 8, characterized in that, Ring A is When R1is -CH2OH, the intermediate of formula (II-2) is first reacted with PBr3to give the intermediate (II-2-a): Then, the intermediate (II-2-a) is reacted with (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H-[1,4]azepino[3,4-c]pyrido[2,1-f][1,2,4]triazine-6,8-dione in the presence of CsF to obtain the intermediate of formula (II-3).

10. The method for synthesizing the pyridone derivative according to claim 8, wherein: when the intermediate (I-1) is of formula (I-1-1), the intermediate (I-1) is obtained by reacting intermediate (I-1-1-1) with tert-butyl nitrite: when the intermediate (I-1) is of formula (I-1-2), the intermediate (I-1) is obtained by reacting intermediate (I-1-2-1) with tert-butyl nitrite: when the intermediate (I-1) is of formula (I-1-3), the intermediate (I-1) is obtained by reacting intermediate (I-1-3-1) in the presence of polyphosphoric acid: when the intermediate (II-1) is of formula (II-1-1), the intermediate (II-1) is obtained by reacting intermediate (II-1-1-1), phenylacetylene and tert-butyl nitrite: when the intermediate (II-1) is of formula (II-1-2), the intermediate (II-1) is obtained by reacting intermediate (II-1-2-1) with the corresponding Grignard reagent of formula (II-1-2-2) in the presence of I: When the intermediate (III-1) is of the structure shown in formula (III-1-1), the intermediate (III-1) is obtained by first reacting intermediate (III-1-1-1) with thionyl chloride, and then reacting under the action of aluminum trichloride: wherein Z1 is S or O; Z2 is CH or N, X1 is S or Se; R2 is selected from 11. Use of the pyridone derivative according to any one of claims 1-5 in the preparation of an influenza drug.

12. The use according to claim 11, wherein the influenza drug is an anti-influenza A drug, an anti-influenza B drug or an anti-avian influenza drug.

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