Pyridone compound, synthetic method and application thereof

Abstract

The invention discloses a pyridone compound as well as a synthesis method and application thereof. The invention discloses a pyridone derivative which has a structure as shown in a general formula I, a general formula II or a general formula III, the invention also discloses a preparation method and the like of the derivative. Researches show that the drug resistance index (RI = IC50 [I38T] / IC50 [WT]) of the hydroxypyridone derivative disclosed by the invention in RNA (Ribonucleic Acid) polymerase of wild type and PA / I38T mutant type influenza viruses is smaller than that of baloxvir, which indicates that the derivative disclosed by the invention is less influenced by the influenza A PA / I38T mutant type and is superior to the baloxvir. Moreover, the cytotoxicity of the derivative is lower than that of baloxvir, so that the derivative has relatively high selectivity and can be applied to preparation of anti-influenza drugs for inhibiting baloxvir drug-resistant virus strains. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the field of medicinal chemistry, and specifically relates to a pyridone compound, a synthesis method and application thereof. Background Art

[0002] Influenza spreads rapidly among the population, with high morbidity and mortality rates. Antiviral treatment for influenza mainly relies on neuraminidase inhibitors, but drug resistance has emerged, and antiviral drugs with new mechanisms are urgently needed. In 2018, the influenza virus cap-dependent nuclease inhibitor baloxavir marboxil was approved for the treatment of acute uncomplicated influenza, with a completely new mechanism of action. However, in recent clinical studies, baloxavir has produced a variety of different mutant virus strains. Among them, PA / I38X (I38T / M / F / L / N / S mutation) mutations account for as much as 2%-9% of patients. Conventional doses of baloxavir treatment are difficult to produce the expected therapeutic effect on influenza patients carrying the I38X mutation. There is an urgent need to develop PA inhibitors with the same level of inhibitory efficacy against wild-type and mutant influenza virus strains. The reduced sensitivity of BXA to the I38T mutation is attributed to unfavorable van der Waals stacking and induced adaptive changes in the Thr38 residue. The butterfly-shaped hydrophobic fragment of BXA is PA N The hydrophobic pocket in the active site provides nearly perfect packing. However, the packing effect is slightly reduced with the presence of Thr38, which is more polar and smaller in size. N The change in binding mode caused by the point mutation of the hydrophobic pocket in the active site, the development of new hydrophobic fragment substituents, and then the weakening of the interaction between the corresponding inhibitor and the I38T mutation site are expected to improve its inhibitory activity against influenza PA / I38T mutant strains. Summary of the invention

[0003] The object of the present invention is to provide a pyridone derivative, which is a hydroxypyridone derivative having 5' cap structure (CAP)-dependent endo-enzyme inhibitory activity.

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

[0005] The present invention further provides an application of the compound in preparing drugs for inhibiting influenza virus RNA polymerase activity and resisting influenza virus.

[0006] Specifically, the structures represented by the following formula (I), formula (II) and formula (III) or their pharmaceutically acceptable salts,

[0007]

[0008] in

[0009] R1 is selected from hydrogen, halogen, alkyl (preferably C1-C6 alkyl), R1 is one or multiple independent of each other;

[0010] R2 is selected from the group consisting of absent, alkyl (preferably C1-C6 alkyl) or alkanoyl (preferably C1-C3 alkanoyl, more preferably )or

[0011] R3 is selected from hydrogen or

[0012] R4 is selected from hydrogen or alkyl (preferably C1-C6 alkyl);

[0013] X is selected from N, O, S, Se; when X is O, S, Se, R2 does not exist;

[0014] Ring A is selected from one of the following structures:

[0015]

[0016] In the present invention, the alkyl group mentioned is generally a C1-C6 alkyl group, including but not limited to methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, pentyl, hexyl, etc. The alkyl group is further preferably a C1-C5 alkyl group, and further preferably a C1-C3 alkyl group, including but not limited to methyl, ethyl, and propyl.

[0017] In the present invention, the halogen mentioned includes F, Cl, Br, and I. Preferably, the halogen includes F, Cl, and Br.

[0018] Furthermore, the C atom corresponding to * in the following formula (I), formula (II) and formula (III) is in S configuration or R configuration or a mixture of S configuration and R configuration:

[0019]

[0020] R1, R2, R3, R4 and X are as defined above.

[0021] Furthermore, R1 is one, two or three independent of each other, and each R1 is independently selected from hydrogen, F, Cl, methyl and ethyl.

[0022] Further, R2 is selected from not existing, or

[0023] Further, R3 is selected from hydrogen or

[0024] Further, R4 is selected from hydrogen or methyl, ethyl;

[0025] Furthermore, X is selected from N, O, S or Se; when X is O, S or Se, R2 does not exist.

[0026] Furthermore, the pyridone derivative has the structure shown in the following formula (Ia), (Ib), (Ic), (III-a):

[0027]

[0028] R1 is H, halogen, C1-C3 alkyl, R1 is 1 or 2 independent of each other, R3 is H or

[0029] Furthermore, the pyridone derivative has a structure shown in any of the following formulae:

[0030]

[0031]

[0032] An intermediate for preparing the pyridone derivative described in any of the above technical solutions has a structure shown in any of the following formulas:

[0033]

[0034] R1 is selected from hydrogen, halogen, alkyl (preferably C1-C6 alkyl), R1 is one or multiple independent of each other;

[0035] R2 is selected from the group consisting of: absent, alkyl (preferably C1-C6 alkyl) or alkanoyl (preferably C1-C3 alkane, more preferably )or

[0036] R4 is selected from hydrogen or (preferably C1-C6 alkyl) alkyl;

[0037] X is selected from N, O, S or Se; when X is O, S or Se, R2 is absent;

[0038] Ring A is selected from one of the following structures:

[0039]

[0040] Furthermore, the intermediate has a structure shown in any of the following formulas:

[0041]

[0042]

[0043] A method for synthesizing a pyridone derivative according to any of the above technical solutions, comprising: reducing a compound represented by formula (I-1) or formula (II-1) or formula (III-1) under the action of sodium borohydride to obtain an intermediate represented by formula (I-2) or formula (II-2) or formula (III-2); reacting the intermediate (I-2) or (II-2) or (III-2) with (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H-[1,4]azino[3,4-c]pyrido[2,1-f][1,2,4]triazine-6,8-dione in the presence of n-butylphosphoric anhydride to obtain an intermediate (I-3) or (II-3) or (III-3); and reducing the intermediate (I-3) or (II-3) or (III-3) with LiCl to obtain a pyridone derivative represented by formula (I) or formula (II) or formula (III):

[0044]

[0045] When preparing the intermediate of formula (I-2) or formula (II-2) or formula (III-2), the solvent used is methanol or ethanol, and the molar ratio of the compound represented by formula (I-1) or formula (II-1) or formula (III-1) to sodium borohydride is 1:1 to 2.5.

[0046] When preparing the intermediate (I-3) or (II-3) or formula (III-3), the solvent used is ethyl acetate; the reaction temperature is 70 to 120°C; further the reaction temperature is 100 to 120°C. The molar ratio of the intermediate (I-2) or (II-2) or formula (III-2) to n-butylphosphoric anhydride and (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H-[1,4]azino[3,4-c]pyrido[2,1-f][1,2,4]triazine-6,8-dione is 1:1 to 1.5:0.3 to 0.6.

[0047] When preparing the pyridone derivatives represented by formula (I) or formula (II) or formula (III), the solvent used is N,N-dimethylacetamide; the reaction temperature is 60 to 100°C; more preferably 70 to 90°C; the molar ratio of the intermediate (I-3) or (II-3) formula (III-3) to LiCl is 1:2 to 15; further preferably 1:5 to 12.

[0048] Furthermore, Ring A For or When the intermediate shown in formula (II-2) first reacts with PBr3 to obtain intermediate (II-2-a):

[0049] Then, the intermediate (II-2-a) reacts with (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H-[1,4]azino[3,4-c]pyrido[2,1-f][1,2,4]triazine-6,8-dione in the presence of CsF to obtain the intermediate represented by formula (II-3).

[0050] In the above steps, when preparing the intermediate (II-2-a), the molar ratio of the intermediate represented by formula (II-2) and PBr3 is 1:1.2~2; the reaction solvent is anhydrous DCM.

[0051] When preparing the intermediate shown in (II-3), the reaction solvent is anhydrous acetonitrile; the molar ratio of (II-2-a) to Cs and (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H-[1,4]azino[3,4-c]pyrido[2,1-f][1,2,4]triazine-6,8-dione is 1:

[0052] 1.5~2.5:0.7~1.2; the reaction temperature is 50~80℃.

[0053] Further:

[0054] When the intermediate (I-1) has a structure represented by formula (I-1-1), the intermediate (I-1) is obtained by reacting the intermediate (I-1-1-1) with tert-butyl nitrite; preferably, the reaction conditions of this step are: the reaction temperature is 70 to 100°C, the reaction solvent is DMSO or the like; the molar ratio of the intermediate (I-1-1-1) to tert-butyl nitrite is 1:1.5 to 4.

[0055] When the intermediate (I-1) is a structure represented by formula (I-1-2), the intermediate (I-1) is obtained by reacting the intermediate (I-1-2-1) with tert-butyl nitrite:

[0056] When the intermediate (I-1) is a structure represented by formula (I-1-3), the intermediate (I-1) is obtained by reacting the intermediate (I-1-3-1) with polyphosphoric acid:

[0057] Preferably, the reaction conditions of this step are: the reaction temperature is 70-100° C., the reaction solvent is DMSO or the like; and the molar ratio of the intermediate (I-1-2-1) to tert-butyl nitrite is 1:1.5-4.

[0058] When the intermediate (II-1) is a structure represented by formula (II-1-1), the intermediate (II-1) is obtained by reacting the intermediate (II-1-1-1), phenylacetylene and tert-butyl nitrite:

[0059] Preferably, the reaction conditions of this step are: the reaction temperature is 50-80°C, the reaction solvent is chloroform or the like; sodium acetate is added at the same time; the molar ratio of the intermediate (II-1-1-1), phenylacetylene, sodium acetate and tert-butyl nitrite is 1:1-1.5:1-1.5:1.5-4.

[0060] When the intermediate (II-1) is a structure represented by formula (II-1-2), the intermediate (II-1) is obtained by reacting the intermediate (II-1-2-1) with a Grignard reagent corresponding to formula (II-1-2-2) under the action of I:

[0061] Preferably, the reaction conditions of this step are: the reaction solvent is anhydrous THF or the like; the molar ratio of the intermediate (II-1-2-1) and the Grignard reagent corresponding to formula (II-1-2-2) is 1:1 to 2.

[0062] When the intermediate (III-1) is a structure represented by formula (III-1-1), the intermediate (III-1) is obtained by first reacting the intermediate (III-1-1-1) with thionyl chloride and then reacting with aluminum chloride:

[0063]

[0064]

[0065] Wherein, Z1 is S or O; Z2 is CH or N, X1 is S or Se; R2 is selected from Specifically:

[0066] The preparation method of the compound corresponding to formula (I) is as follows: X is O, R3 is H, and R2 does not exist;

[0067]

[0068] Or the preparation method is as follows: X is N, R3 is H;

[0069]

[0070] The preparation method of the compound corresponding to formula (I) is as follows: X is S, R3 is H, and R2 does not exist;

[0071]

[0072] The preparation method of the compound corresponding to formula (I) is as follows: X is Se, R3 is H, and R2 does not exist;

[0073]

[0074] The preparation method of the compound corresponding to formula (II) is as follows: R3 is H;

[0075]

[0076] Or the preparation method of the compound corresponding to formula (II) is as follows:

[0077]

[0078] Or the preparation method of the compound corresponding to formula (III) is as follows:

[0079]

[0080] The present invention also provides a use of any of the above-mentioned pyridone derivatives in the preparation of anti-influenza A drugs.

[0081] The positive and progressive effects of the present invention are:

[0082] The hydroxypyridone compounds of the present invention can effectively inhibit the replication of influenza virus in vitro. In particular, the compounds of the present invention are less affected by influenza PA / I38T mutant strains, and the drug resistance index (RI=IC 50 [I38T] / IC 50 [WT]) is significantly lower than that of positive baloxavir. The present invention also discloses the use of the hydroxypyridone compound in the preparation of an anti-baloxavir resistant virus drug, which has low cytotoxicity and high selectivity. The hydroxypyridone compound with a new structure disclosed in the present invention has a good application prospect in the clinical treatment of baloxavir resistant virus strains. DETAILED DESCRIPTION

[0083] The present invention is further described in detail below in conjunction with embodiments, but the embodiments of the present invention are not limited thereto. Without departing from the above-mentioned technical concept of the present invention, various substitutions and changes are made according to common technical knowledge and customary means in the field, which should all be included in the scope of the present invention.

[0084] Example 1. Synthesis of 1-6-A and 1-6-B

[0085]

[0086] Step 1: Synthesis of compound 1-2

[0087] Under ice bath, potassium carbonate (8.1g, 58.8mmol) was added to a DMF solution (10mL) of compound 1-1 (5.0g, 29.4mmol), and then 3-bromopropyne (3.0mL, 44.2mmol) was added dropwise under stirring. After the addition was completed, the reaction solution was restored to room temperature and heated to 60°C, and stirred for 12h. After the reaction was completed, the reaction solution was added to water (100mL), and extracted with ethyl acetate (3×100mL), the organic phases were combined, and the organic phases were washed with water (1×100mL) and saturated brine (1×100mL) in turn, dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude compound 1-2. Finally, the pure compound 1-2 (5.6g, yield 92%) was obtained by recrystallization from petroleum ether / ethyl acetate. 1 H NMR (500MHz, CDCl3) δ7.72(d,J=8.3Hz,1H),7.07(d,J=1.8Hz,1H),7.04(dd,J=8.4,1 .8Hz,1H),4.80(d,J=2.4Hz,2H),2.61(s,3H),2.59(t,J=2.4Hz,1H).HRMS(ESI):m / z calcd for C 11 H 10 ClO2 + [M+H] + :209.0364,found:209.0360.

[0088] Step 2: Synthesis of compound 1-3

[0089] In a sealed tube, compound 1-2 (4.16 g, 20 mmol) was dissolved in a DMSO solution (5 mL), and then tert-butyl nitrite (7.1 mL, 60 mmol) was added dropwise under stirring. After the addition was completed, the reaction solution was heated to 80°C and stirred for 4 hours. After the reaction was completed, the reaction solution was added to water (50 mL) and extracted with ethyl acetate (3×50 mL). The organic phases were combined, and the organic phases were washed with water (1×100 mL) and saturated brine (1×100 mL) in sequence, dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The obtained crude product was purified by silica gel column chromatography to obtain pure compound 1-3 (2.35 g, 50%). 1 H NMR (500MHz, DMSO-d6) δ9.21 (s, 1H), 8.13 (d, J = 8.7Hz, 1H), 7.37 (dd, J = 8.7, 2.1Hz, 1H), 7.30 (d, J = 2.1Hz, 1H), 5.28 (s, 2H). HRMS (ESI): m / z calcd for C 11 H7ClNO3 + [M+H]+ :236.0109,found:236.0101

[0090] Step 3: Synthesis of Compound 1-4

[0091] In an ice bath, compound 1-3 (2.35 g, 10 mmol) was added to 15 mL of methanol, and then sodium borohydride (0.76 g, 20 mmol) was slowly added under stirring, and the reaction solution was warmed to room temperature and continued to stir for 2 h. After the reaction was completed, 5 mL of water was added to quench the reaction, and then most of the methanol was removed by concentration under reduced pressure. The aqueous phase was extracted with dichloromethane (2 × 50 mL), and the organic phases were combined and washed with saturated brine (1 × 50 mL), dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain the crude product of 1-4. Finally, the pure product of compound 1-4 (2.18 g, yield 92%) was obtained by recrystallization from ethyl acetate. 1 H NMR(500MHz,DMSO-d6)δ8.74(d,J=1.2Hz,1H),7.52(dd,J=8.1,0.7Hz,1H),7.26–7.09( m,2H),6.44(d,J=5.4Hz,1H),5.93(d,J=5.3Hz,1H),5.21–5.11(m,2H).HRMS(ESI):m / z calcd for C 11 H7ClNO2 + [M-OH] + :220.0160,found:220.0162

[0092] Step 4: Synthesis of compound 1-5

[0093] Compound 1-4 (0.28 g, 1.2 mmol) was added to a 50% (w / w)-T4P / EA (1 mL, 1.2 mmol) solution (n-butyl phosphoric acid (50% ethyl acetate solution)), and the mixture was stirred at room temperature. Then (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H-[1,4]azino[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 solution was heated to 110°C and stirred for 5 hours. After the reaction was completed, the reaction solution was added to 15 mL of a saturated sodium bicarbonate solution for quenching and extracted with ethyl acetate (2×50 mL). After the organic phases were combined, they were washed with saturated brine (1×50 mL), dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The crude product was purified by column chromatography to obtain 58 mg and 52 mg of pure compounds 1-5-A and 1-5-B, respectively (total yield 17%). The obtained compounds were detected by liquid chromatography column, and the retention time of compound 1-5-A was 7.10 min, and the retention time of compound 1-5-B was 7.14 min. Compound 1-5-A and compound 1-5-B are two non-corresponding isomers of the above formula (1-5) with respect to *C, the same below.

[0094] Compound 1-5-A: 1 H NMR(500MHz, CDCl3)δ8.28(s,1H),7.67–7.64(m,2H),7.39(dd,J=8.0,6.4Hz,2H),7.37–7.28(m,3H),7.26(s ,1H),6.98(d,J=7.6Hz,1H),6.13(d,J=7.8Hz,1H),5.58(s,1H),5.53–5.46(m,2H),5.38(d,J=10.6Hz,1H),4 .87(dd,J=14.8,1.4Hz,1H),4.65(dd,J=13.6,2.6Hz,1H),4.50(dd,J=9.9,3.0Hz,1H),3.83(dd,J=10.8,3.0 Hz,1H),3.75(dd,J=11.9,3.3Hz,1H),3.33–3.24(m,2H),2.99(ddd,J=13.4,11.8,3.5Hz,1H).HRMS(ESI):m / z calcd for C 28 H 24 ClN4O6 + [M+H] + :547.1379,found:547.1371Compound 1-5-B: 1H NMR(500MHz, CDCl3)δ8.30(s,1H),7.61–7.56(m,2H),7.38–7.33(m,2H),7.32–7.28(m,1H),7.12(d,J=2.1Hz ,1H),6.96(d,J=7.7Hz,1H),6.78(dd,J=8.1,2.1Hz,1H),6.44(d,J=8.2Hz,1H),5.79(d,J=7.7Hz,1H),5.59(d ,J=11.0Hz,1H),5.48–5.43(m,2H),5.41(d,J=11.0Hz,1H),4.87(dd,J=14.7,1.3Hz,1H),4.79(dd,J=9.9,3.0 Hz,1H),4.70(dd,J=13.3,2.3Hz,1H),3.80(ddd,J=19.6,11.2,3.1Hz,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

[0095] Step 5: Synthesis of compound 1-6-A

[0096] Compound 1-5-A (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 solution 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 HCl aqueous solution and 1 mL of water were added, and stirring was continued for 1 h. The solid precipitated from the reaction solution was collected by suction filtration and dried to obtain a crude compound 1-6-A. Further recrystallization from isopropyl ether / chloroform gave a pure compound 1-6-A (36 mg, yield 79%). White solid, mp 212.4-213.3 ° C. 1H NMR (500MHz, DMSO-d6) δ8.95(s,1H),7.37(d,J=9.5Hz,1H),7.25(d,J=2.2Hz,1H),7.10(d,J= 8.3Hz,1H),7.05(d,J=7.6Hz,1H),5.92(s,1H),5.66(d,J=7.6Hz,1H),5.49(d,J=14.8Hz,1H), 4.94(d,J=14.8Hz,1H),4.85(d,J=9.8Hz,1H),4.43(d,J=13.4Hz,1H),3.77(dd,J=10.8,3.1H z,1H),3.73(dd,J=11.7,3.3Hz,1H),3.62–3.55(m,1H),3.40-3.42(m,1H),3.16–3.10(m,1H). 13 C NMR(125MHz,DMSO-d6)δ160.93,158.20,157.12,157.06,138.20,135.71,133.67,130.81,125.24, 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

[0097] Step 6: Synthesis of compound 1-6-B

[0098] 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 solution 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 HCl aqueous solution and 1 mL of water were added, and stirring was continued for 1 h. The solid precipitated from the reaction solution was collected by suction filtration and dried to obtain a crude compound 1-6-B. Further recrystallization from isopropyl ether / chloroform gave a pure compound 1-6-B (30 mg, yield 66%). White solid, mp 204.1-205.0 ° C. 1H NMR (500MHz, DMSO-d6) δ11.66(s,1H),8.80(s,1H),7.68(d,J=8.2Hz,1H),7.38(d,J=2.1 Hz,1H),7.32(dd,J=8.2,2.2Hz,1H),7.18(d,J=7.6Hz,1H),5.84(s,1H),5.76(d,J=7.7Hz ,1H),5.55(d,J=14.9Hz,1H),4.91(d,J=14.9Hz,1H),4.53–4.45(m,1H),4.42(d,J=13.4 Hz,1H),3.68(td,J=10.5,3.2Hz,2H),3.63–3.55(m,1H),3.39(s,1H),3.15–3.09(m,1H). 13 C NMR(125MHz,DMSO-d6)δ160.87,158.39,156.34,155.62,138.55,135.71,133.40,126.79,125.20,124.02,1 16.14,111.10,79.26,79.00,78.74,70.27,68.06,65.54,64.73,61.97,45.37,22.79.HRMS(ESI):m / zcalcd for C 21 H 18 ClN4O6 + [M+H] + :457.0910,found:457.0910

[0099] Example 2. Synthesis of 2-6-A and 2-6-B

[0100] Referring to the method of Example 1, except that 4'-chloro-2'-hydroxyacetophenone (1-1) was replaced by 4-fluoro-2-hydroxyacetophenone, two diastereoisomers corresponding to compound 2-6 were finally obtained: compound 2-6-A and compound 2-6-B. Compound 2-6-A: white solid, mp 226.9-227.3℃. 11H 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 13C 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 1H 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 (125MHz, DMSO-d6) δ171.31,160.88,159.11(d,J=11.8Hz),156.33,155.85,152.81,138.57,133.63(d,J=10.58Hz),116.13, 115.86,112.13(d,J=21.50Hz),111.40(d,J=22.68Hz),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

[0101] Example 3. Synthesis of 3-6-A and 3-6-B

[0102] Referring to the method of Example 1, except that 4'-chloro-2'-hydroxyacetophenone (1-1) was replaced with 2'-hydroxyacetophenone, two diastereoisomers corresponding to compound 3-6 were finally obtained: compound 3-6-A and compound 3-6-B.

[0103] Compound 3-6-A: white solid, mp 228.3-229.5℃. 1 H NMR(500MHz,DMSO-d6)δ8.90(s,1H),7.40(t,J=7.7Hz,1H),7.31(d,J=7.5Hz,1H),7.15(d,J=7.9Hz,1 H),7.08(d,J=7.7Hz,1H),7.02(t,J=7.3Hz,1H),5.84(s,1H),5.65(d,J=7.6Hz,1H),5.46(d,J=14.9H z,1H),4.93(dd,J=10.0,3.2Hz,1H),4.87–4.81(m,1H),4.43(dd,J=13.4,2.6Hz,1H),3.75(ddd,J=17 .9,11.2,3.2Hz,2H),3.60(d,J=10.4Hz,1H),3.42–3.39(m,1H),3.16(ddd,J=15.0,11.9,3.5Hz,1H). 1313C 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

[0104] Compound 3-6-B: White solid, mp 215.1 - 216.1 °C. 1 1H 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 13C 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

[0105] Example 4. Synthesis of 4-9-A and 4-9-B

[0106]

[0107] Step 1: Synthesis of compound 4-2

[0108] At room temperature, di-tert-butyl dicarbonate (9.3 mL, 40.7 mmol) was added to an ethanol solution (15 mL) of compound 4-1 (5.0 g, 37.0 mmol). After the addition was complete, the reaction solution was heated to 65°C and stirred for 24 hours. After the reaction was completed, the reaction solution was dried to obtain a crude compound 4-2 (7.4 g, yield 85%), which could be directly used in the next step. 1 H NMR(500MHz, CDCl3) δ10.93(s,1H),8.46(dd,J=8.6,1.2Hz,1H),7.85(dd,J=8.0,1.6Hz,1H),7 .54–7.46(m,1H),7.02(ddd,J=8.2,7.2,1.2Hz,1H),2.64(s,3H),1.52(s,9H).HRMS(ESI):m / z calcdfor C 13 H 17 NaNO3 + [M+Na] + :258.1101,found:258.1109

[0109] Step 2: Synthesis of compound 4-3

[0110] Under an ice bath, NaH (1.44 g, 60 mmol) was added in batches to 30 mL of anhydrous THF solution of compound 4-2 (7.05 g, 30 mmol) under N2 protection conditions, and then returned to room temperature and stirred for 1 h. 3-Bromopropyne (3.9 mL, 45 mmol) was added dropwise to the reaction solution, and the reaction was refluxed overnight. After the reaction was completed, 10 mL of saturated NH4Cl solution was added to quench the reaction, and then most of the THF was removed by concentration under reduced pressure. The aqueous phase was extracted with ethyl acetate (2 × 50 mL), and the organic phases were combined and washed with saturated brine (1 × 50 mL), dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude product of 4-3. The obtained crude product was purified by silica gel column chromatography to obtain a pure product of 4-3 (5.0 g, yield 61%). 1 H NMR (500MHz, CDCl3) δ7.64 (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 = 95.1Hz, 9H). HRMS (ESI): m / z calcd forC 16H 19 NaNO3 + [M+Na] + :296.1258,found:296.1257

[0111] Step 3: Synthesis of compound 4-4

[0112] In a sealed tube, compound 4-3 (4.1 g, 15 mmol) was dissolved in a DMSO solution (5 mL), and then tert-butyl nitrite (5.3 mL, 45 mmol) was added dropwise under stirring. After the addition was completed, the reaction solution was heated to 80°C and stirred for 4 hours. After the reaction was completed, the reaction solution was added to water (50 mL) and extracted with ethyl acetate (3×50 mL). The organic phases were combined and washed with saturated brine (1×100 mL), dried over anhydrous sodium sulfate, filtered, and spin-dried. The obtained crude product was purified by silica gel column chromatography to obtain pure compound 4-4 (1.9 g, yield 42%). 1 H NMR (500MHz, CDCl3) δ8.56 (s, 1H), 8.14 (s, 1H), 7.61 (t, J = 7.4Hz, 1H), 7.50–7.28 (m, 2H), 5.70–3.89 (m, 2H), 1.31 (s, 9H). HRMS (ESI): m / z calcdfor C 16 H 16 NaN2O4 + [M+Na] + :323.1003,found:323.1002

[0113] Step 4: Synthesis of compound 4-5

[0114] At room temperature, compound 4-4 (1.5 g, 5 mmol) was dissolved in 10 mL of anhydrous DCM, and then 5 mL of trifluoroacetic acid was added dropwise under stirring. After the addition was completed, the reaction solution was stirred at room temperature for 6 h. After the reaction was completed, the reaction solution was dried, the residue was dissolved with 20 mL of DCM, and a saturated sodium bicarbonate solution was added to adjust the pH value to 7. The aqueous phase was extracted with DCM (3×20 mL), and the organic phases were combined and washed with saturated brine (1×50 mL), dried over anhydrous sodium sulfate, filtered, and dried. The crude product was recrystallized from petroleum ether / dichloromethane to obtain pure compound 4-5 (0.76 g, yield 76%). 1H NMR(500MHz,DMSO-d6)δ9.03(s,1H),7.99(dd,J=8.3,1.7Hz,1H),7.35(ddd,J=8.4,5.2,1.7Hz,2H),6.9 1(dd,J=8.5,1.1Hz,1H),6.78(ddd,J=8.2,6.8,1.1Hz,1H),4.23(d,J=3.8Hz,2H).HRMS(ESI):m / zcalcd for C 11 H9N2O2 + [M+H] + :201.0659,found:201.0653

[0115] Step 5: Synthesis of Compound 4-6

[0116] At room temperature, compound 4-5 (0.76 g, 3.8 mmol) and DIPEA (0.7 mL, 4 mmol) were dissolved in 5 mL of anhydrous DCM, and then acetyl chloride (0.32 mL, 4.5 mmol) was added dropwise under stirring. After the addition was completed, the reaction solution was stirred at room temperature for 2 h. After the reaction was completed, 20 mL of DCM was added to dilute the reaction solution, and then washed with dilute hydrochloric acid (0.5 M, 1 × 20 mL), saturated sodium bicarbonate (1 × 20 mL), and saturated sodium chloride solution (1 × 20 mL), respectively, and the organic phases were combined and dried over anhydrous sodium sulfate, filtered, and spin-dried. The crude product was then slurried with ether to obtain pure compound 4-6 (0.8 g, yield 87%). 1 H NMR (500MHz, CDCl3) δ8.65–8.53(m,1H),8.16(dd,J=7.8,1.7Hz,1H),7.70(dd,J=7.7,1.7Hz,1H),7.59(dd,J=7.7,1.2 Hz,1H),7.35(dd,J=7.9,1.2Hz,1H),5.86(d,J=15.7Hz,1H),4.04(dd,J=15.7,1.5Hz,1H),1.86(s,3H).HRMS(ESI):m / z calcd for C 13 H 11 N2O3 + [M+H] + :243.0795,found:243.0794

[0117] Step 6: Synthesis of Compound 4-7

[0118] Under ice bath, compound 4-6 (0.8 g, 3.3 mmol) was added to 10 mL of methanol, and then sodium borohydride (0.25 g, 6.6 mmol) was slowly added under stirring, and the reaction solution was warmed to room temperature and continued to stir for 2 h. After the reaction was completed, 5 mL of water was added to quench the reaction, and then most of the methanol was removed by concentration under reduced pressure. The aqueous phase was extracted with dichloromethane (2 × 50 mL), and the organic phases were combined and washed with saturated brine (1 × 50 mL), dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain the crude product of 4-7. Finally, the pure product of compound 4-7 (0.72 g, yield 90%) was obtained by recrystallization from ethyl acetate. 1 H NMR (500MHz, DMSO-d6) δ8.72(d,J=1.1Hz,1H),7.58(dd,J=7.5,1.6Hz,1H),7.51–7.39(m,3H),6.17(d,J=141. 5Hz,1H),5.80(dd,J=16.4,1.2Hz,1H),5.72(s,1H),3.81(dd,J=16.4,1.5Hz,1H),1.76(s,3H).HRMS(ESI):m / z calcd for C 13 H 13 N2O3 + [M+H] + :245.0921,found:245.0918

[0119] Step 7: Synthesis of compounds 4-8-A and 4-8-B

[0120] In a sealed tube, compound 4-7 (0.29 g, 1.2 mmol) was added to a 50% (w / w)-T4P / EA (1 mL, 1.2 mmol) solution, and the mixture was stirred at room temperature. Then (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H-[1,4]azino[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 solution was heated to 110°C and stirred for 5 h. After the reaction was completed, the reaction solution was added to 15 mL of saturated sodium bicarbonate solution for quenching and extracted with ethyl acetate (2×50 mL). After the organic phases were combined, they were washed with saturated brine (1×50 mL), dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The crude product was purified by column chromatography to obtain 46 mg and 40 mg of pure compounds 4-8-A and 4-8-B, respectively (total yield 13%). The obtained compounds were detected by liquid chromatography column, and the retention time of compound 4-8-A was 7.60 min, and the retention time of compound 4-8-B was 7.70 min. Compound 4-8-A and compound 4-8-B are two non-corresponding isomers corresponding to the above compound 4-8.

[0121] Compound 4-8-A: 1 H NMR(500MHz,DMSO-d6)δ9.22(s,1H),8.04(dd,J=7.8,1.7Hz,1H),7.80(td,J=7.6,1.7Hz,1H),7.70(d,J=7.6Hz,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=7.6Hz,1H),5.84(s,1H),5.61(m,1H),5.07(d,J=2.4Hz,2H),4.80(ddd,J=12.6,9.9,4.6Hz,1H),4.18–4.10(m,2H),4.03(td,J=11.3,4.4Hz,2H),3.43(td,J=12.0,3.1Hz,1H),3.13(dd,J=11.4,10.0Hz,1H),2.95(ddd,J=13.7,12.1,4.3Hz,1H),1.77(s,3H).HRMS(ESI):m / z calcd for C 30 H 28 N5O6 + [M+H] + :554.2035,found:554.2038

[0122] Compound 4-8-B: 1 H NMR(500MHz,DMSO-d6)δ8.72(d,J=1.1Hz,1H),7.70(d,J=7.6Hz,1H),7.57(td,J=7.8,1.4Hz,3H),7.53–7.39(m,2H),7.38–7.34(m,2H),7.32–7.28(m,2H),6.22(d,J=7.6Hz,1H),5.80(s,1H),5.68(m,1H),5.08(d,J=2.3Hz,2H),4.83–4.77(m,1H),4.15(dd,J=13.8,2.9Hz,1H),4.02(dt,J=11.2,5.4Hz,2H),3.81(dd,J=16.4,1.5Hz,1H),3.46–3.41(m,1H),3.13(dd,J=11.4,9.9Hz,1H),2.95(ddd,J=13.8,12.2,4.3Hz,1H),1.76(s,3H).HRMS(ESI):m / z calcd for C 30 H 28N5O6 + [M+H] + :554.2035,found:554.2040

[0123] Step 8: Synthesis of compound 4-9-A

[0124] Compound 4-8-A (46 mg, 0.08 mmol) was added to 1 mL N, N-dimethylacetamide, lithium chloride (34 mg, 0.8 mmol) was added, and the reaction solution 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 acetone, 2 mL 0.5 M HCl aqueous solution and 1 mL water were added, and stirring was continued for 1 h. The solid precipitated from the reaction solution was collected by suction filtration and dried to obtain a crude compound 4-9-A. Further recrystallization from isopropyl ether / chloroform gave a pure compound 4-9-A (31 mg, yield 83%). White solid, mp236.0-237.0 ° C. 1 H NMR(500MHz, DMSO-d6)δ9.21(s,1H),8.04(dd,J=7.8,1.6Hz,1H),7.80(td,J=7.6,1.6Hz,1H),7.64–7.32(m,3H),6.07(d,J=7.4Hz,1H),5.75(s, 2H),4.88(td,J=13.2,11.7,4.1Hz,1H),4.20–3.95(m,3H),3.5-3.75(m ,1H),3.22(t,J=10.8Hz,2H),3.02(td,J=13.1,4.1Hz,1H),1.77(s,3H). 13 C NMR (125MHz, DMSO-d6) δ182.20,170.72,168.72,162.38,158.63,158.52,151.61,141.43,137.80,135.30,134. 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

[0125] Step 9: Synthesis of compound 4-9-B

[0126] Compound 4-8-B (40 mg, 0.07 mmol) was added to 1 mL N, N-dimethylacetamide, lithium chloride (30 mg, 0.7 mmol) was added, and the reaction solution 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 acetone, 2 mL 0.5 M HCl aqueous solution and 1 mL water were added, and stirring was continued for 1 h. The solid precipitated from the reaction solution was collected by suction filtration and dried to obtain a crude compound 4-9-B. Further recrystallization by isopropyl ether / chloroform gave a pure compound 4-9-B (27 mg, yield 82%). White solid, mp230.6-231.2 ° C. 1 H NMR(500MHz,DMSO-d6)δ8.70(d,J=18.0Hz,1H),7.57(dd,J=7.4,1.5Hz,1H),7.50(td,J=7.5,1 .6Hz,1H),7.43(dtd,J=17.9,7.5,1.5Hz,2H),7.39–7.20(m,1H),6.01(d,J=15.5Hz,1H),5.80 (d,J=16.7Hz,1H),5.74(d,J=19.2Hz,1H),4.82(s,1H),4.12–3.98(m,3H),3.81(d,J=16.4Hz, 1H),3.47–3.42(m,1H),3.21(t,J=10.7Hz,1H),2.97(t,J=12.8Hz,1H),1.77(d,J=8.4Hz,3H). 13 CNMR(125MHz,DMSO-d6)δ169.56,168.79,162.25,161.17,160.56,155.86,141.46,138.41,130.21,130.09,129 .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

[0127] Example 5

[0128]

[0129] Step 1: The synthesis of compounds 5-1 to 5-5 is the same as in Example 4

[0130] Step 2: Synthesis of compound 5-6

[0131] At room temperature, compound 5-5 (1.0 g, 5 mmol) and DIPEA (1.1 mL, 6 mmol) were dissolved in 5 mL of anhydrous DCM, and then chloroacetyl chloride (0.48 mL, 6 mmol) was added dropwise under stirring. After the addition was completed, the reaction solution was stirred at room temperature for 2 h. After the reaction was completed, 20 mL of DCM was added to dilute the reaction solution, and then washed with dilute hydrochloric acid (0.5 M, 1 × 20 mL), saturated sodium bicarbonate (1 × 20 mL), and saturated sodium chloride solution (1 × 20 mL), respectively, and the organic phases were combined and dried over anhydrous sodium sulfate, filtered, and spin-dried. The crude product was then slurried with ether to obtain pure compound 5-6 (1.3 g, yield 93%). 1 H NMR (500MHz, CDCl3) δ8.62(d,J=1.4Hz,1H),8.20(dd,J=7.9,1.7Hz,1H),7.75(td,J=7.6,1.7Hz,1H),7.64(td,J=7.6,1.2Hz,1H),7.46(dd,J =7.8,1.2Hz,1H),5.83(d,J=15.7Hz,1H),4.12(dd,J=15.7,1.5Hz,1H),3.87(d,J=12.8Hz,1H),3.78(d,J=12.8Hz,1H).HRMS(ESI):m / zcalcd for C 13 H 10 N2ClO3 + [M+H] + :277.0375,found:277.0368

[0132] Step 3: Synthesis of Compound 5-7

[0133] At room temperature, compound 5-6 (1.1 g, 4 mmol) was dissolved in 5 mL of acetonitrile, and then KSCN (1.2 g, 12 mmol) was added under stirring, and the reaction solution was stirred at 60 ° C for 4 h. After the reaction was completed, the solvent was dried and dissolved with 20 mL of ethyl acetate, and the organic phase was washed with water (1×50 mL), saturated brine (1×50 mL), dried over anhydrous sodium sulfate, filtered, and dried to obtain a crude product. Finally, ether was used to slurry to obtain the pure compound 5-7 (1.0 g, yield 88%). 1H NMR (500MHz, DMSO-d6) δ9.23(s,1H),8.11(dd,J=7.9,1.7Hz,1H),7.80(td,J=7.6,1.7Hz,1H),7.72(dd,J=8.0,1.3Hz,1H),7.66(td,J =7.6,1.3Hz,1H),5.60(d,J=15.7Hz,1H),4.42(d,J=15.1Hz,1H),4.24(dd,J=15.7,1.5Hz,1H),3.85(d,J=15.1Hz,1H).HRMS(ESI):m / z calcd forC 14 H 10 N3SO3 + [M+H] + :300.0438,found:300.0430

[0134] Step 4: The synthesis of compounds 5-8 to 5-10-A and 5-10-B is the same as in Example 4

[0135] Compound 5-10-A: white solid, mp 239.0-240.0℃. 1 H NMR(500MHz,DMSO-d6)δ9.23(s,1H),8.11(dd,J=7.8,1.5Hz,1H),7.80(t,J=7.5Hz,1H),7.72(d, J=7.8Hz,1H),7.65–7.61(m,1H),7.33(d,J=12.5Hz,1H),6.07(d,J=7.4Hz,1H),5.88–5.48(m,2H) ,4.89(ddd,J=13.5,10.1,4.0Hz,1H),4.41(d,J=15.1Hz,1H),4.24(d,J=15.4Hz,1H),4.17–3.99 (m,4H),3.84(d,J=15.0Hz,1H),3.46(d,J=2.9Hz,1H),3.22(s,1H),3.01(dd,J=13.2,4.2Hz,1H). 13C NMR(125MHz,DMSO-d6)δ181.42,169.65,165.44,162.25,158.99,158.78,151.38,139.79,138.35,135.75,134.69, 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

[0136] Compound 5-10-B: white solid, mp 245.1-246.0℃. 1 H NMR(500MHz,DMSO-d6)δ8.69(s,1H),7.66–7.60(m,2H),7.50(dd,J=6.9,2.1Hz,1H),7.43(ddd,J =7.0,4.8,1.9Hz,2H),7.41–7.32(m,1H),6.12(s,1H),6.07(d,J=7.4Hz,1H),4.89(td,J=15.7,14 .5,8.5Hz,2H),4.48(d,J=16.8Hz,1H),4.09(dd,J=14.1,2.9Hz,1H),4.03(td,J=11.7,5.5Hz,2H) ,3.47(dd,J=12.2,2.9Hz,1H),3.28(s,2H),3.23(t,J=10.7Hz,1H),3.02(td,J=13.1,4.1Hz,1H). 13 C NMR(125MHz,DMSO-d6)δ170.62,167.50,162.35,161.22,155.61,151.56,141.62,137.73,136.71,129.42,128.98, 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

[0137] Example 6. Synthesis of 6-6-A and 6-6-B

[0138]

[0139] Step 1: Synthesis of compound 6-2

[0140] At room temperature, compound 6-1 (5.9 g, 30 mmol) was dissolved in 20 mL of acetone, dimethyl sulfide (3.3 mL, 45 mmol) was added, and the reaction solution was stirred at room temperature overnight. After the reaction was completed, the reaction solution was filtered to obtain a white solid, which was dried to obtain a crude compound 6-2 (7.0 g, yield 90%), which was directly used in the next step.

[0141] Step 2: Synthesis of compound 6-3

[0142] In a sealed tube, compound 6-2 (5.2 g, 20 mmol) was added to 10 mL of chloroform, followed by sodium acetate (1.64 g, 20 mmol), phenylacetylene (2.2 mL, 20 mmol), and tert-butyl nitrite (7.1 mL, 60 mmol), and the reaction solution was heated to 60 ° C and stirred for 6 h. After the reaction was completed, most of the solvent was removed by rotary evaporation, and the residue was dissolved in 20 mL of DCM. The organic phases were combined and washed with water (1×50 mL), saturated brine (1×50 mL), dried over anhydrous sodium sulfate, filtered, and dried to obtain a crude product, which was purified by silica gel column chromatography to obtain pure compound 6-3 (3.3 g, yield 66%). 1 H NMR (500MHz, CDCl3) δ8.38–8.33(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

[0143] Step 3: Synthesis of compound 6-4

[0144] Under ice bath, compound 6-3 (2.5 g, 10 mmol) was added to 10 mL of methanol, and then sodium borohydride (0.76 g, 20 mmol) was slowly added under stirring, and the reaction solution was warmed to room temperature and continued to stir for 2 h. After the reaction was completed, 5 mL of water was added to quench the reaction, and then most of the methanol was removed by concentration under reduced pressure. The aqueous phase was extracted with dichloromethane (2 × 50 mL), and the organic phases were combined and washed with saturated brine (1 × 50 mL), dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain the crude product of 6-4. Finally, the pure product of compound 6-4 (2.2 g, yield 90%) was obtained by recrystallization from petroleum ether / ethyl acetate. 1 H NMR(500MHz, CDCl3)δ7.75–7.69(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=2.1Hz,1H),3.02(d,J=3.3Hz,1H).HRMS(ESI):m / z calcd for C 16 H 12 NO + [M-OH] + :234.0914,found:234.0918

[0145] Step 4: Synthesis of compounds 6-5-A and 6-5-B

[0146] In a sealed tube, compound 6-4 (0.3 g, 1.2 mmol) was added to a 50% (w / w)-T4P / EA (1 mL, 1.2 mmol) solution, and the mixture was stirred at room temperature. Then (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H-[1,4]azino[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 solution was heated to 110°C and stirred for 5 h. After the reaction was completed, the reaction solution was added to 15 mL of saturated sodium bicarbonate solution for quenching and extracted with ethyl acetate (2×50 mL). After the organic phases were combined, they were washed with saturated brine (1×50 mL), dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The crude product was purified by column chromatography to obtain 30 mg and 24 mg of pure compounds 6-5-A and 6-5-B, respectively (total yield 8%). The obtained compounds were detected by liquid chromatography column, and the retention time of compound 6-5-A was 7.0 min, and the retention time of compound 6-5-B was 7.1 min.

[0147] Compound 6-5-A: 11H 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

[0148] Compound 6-5-B: 1 1H 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

[0149] Step 5: Synthesis of compound 6-6-A

[0150] Compound 6-5-A (30 mg, 0.053 mmol) was added to 1 mL of N, N-dimethylacetamide, and lithium chloride (26 mg, 0.6 mmol) was added. The reaction solution was stirred at 80 ° C for 6 hours. 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 HCl aqueous solution and 1 mL of water were added, and stirring was continued for 1 hour. The solid precipitated from the reaction solution was collected by suction filtration and dried to obtain a crude product. Further recrystallization from isopropyl ether / chloroform gave the pure compound 6-6 (17 mg, yield 68%). White solid, mp 198.0-199.0 ° C 1 H NMR(500MHz,DMSO-d6)δ7.87–7.84(m,2H),7.62(d,J=7.5Hz,1H),7.51–7.46(m ,5H),7.36(q,J=6.9,6.2Hz,3H),7.28(t,J=7.3Hz,1H),6.98(s,1H),6.07(d,J= 7.4Hz,1H),5.85(s,1H),4.93–4.84(m,1H),4.09(dd,J=14.1,3.0Hz,1H),4.03 (td,J=11.7,5.5Hz,2H),3.46(d,J=2.9Hz,1H),3.23(s,1H),3.05–2.99(m,1H). 13 C NMR (125MHz, DMSO-d6) δ169.40,168.56,162.68,162.45,151.96,142.57,138.35,131.00,129.80,128. 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

[0151] Step 6: Synthesis of compound 6-6-B

[0152] Compound 6-5-B (24 mg, 0.043 mmol) was added to 1 mL N, N-dimethylacetamide, lithium chloride (26 mg, 0.6 mmol) was added, and the reaction solution 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 acetone, 2 mL 0.5 M HCL aqueous solution and 1 mL 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 a crude product. Further recrystallization from isopropyl ether / chloroform gave the pure compound 6-6-B (13 mg, yield 64%). White solid, mp 202.5-203.1 ° C.. 1 H NMR(500MHz,DMSO-d6)δ8.20(d,J=7.6Hz,2H),8.02(dd,J=7.3,2.1Hz,1H),7.78(t,J=7.4Hz,1 H),7.73(dd,J=7.6,1.6Hz,1H),7.66–7.61(m,4H),7.60–7.56(m,1H),7.50(s,1H),7.35(d,J= 12.3Hz,1H),6.07(d,J=7.4Hz,1H),5.75(s,1H),4.92–4.85(m,1H),4.08(td,J=13.3,12.7,4. 9Hz, 2H), 4.03–4.00 (m, 1H), 3.45 (d, J = 2.9Hz, 1H), 3.24 (d, J = 10.7Hz, 1H), 3.04–2.98 (m, 1H). 13 C NMR (125MHz, DMSO-d6) δ185.40,170.62,167.14,162.34,161.89,151.56,151.08,137.72,135.28,134.49,132.49,131.12,1 31.01,130.25,130.08,128.89,128.07,124.84,118.21,111.72,104.92,99.76,65.50,65.44,64.74,54.91.HRMS(ESI):m / z calcd for C 26 H 23 N4O5 + [M+H] + :471.1663,found:471.1658

[0153] Example 7. Synthesis of 7-6-A and 7-6-B

[0154] The method of Example 6 was followed except that 2-bromoacetophenone (6-1) was replaced by α-bromo-4-chloroacetophenone. The total yield was 9%.

[0155] Compound 7-6-A: white solid, mp 186.2-187.2℃. 1 H NMR (500MHz, DMSO-d6) δ8.23(m,2H),8.00(dd,J=7.6,1.8Hz,3H),7.70(d,J=8.6Hz,3H),7.58(m,2H),7.35(d,J=12.4Hz,1H),6. 07(d,J=7.4Hz,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=10.7Hz,1H),3.04–2.99(m,1H). 13 C NMR (125MHz, DMSO-d6) δ185.69,170.46,169.44,164.53,162.54,162.32,162.27,151.52,137.79,135.39,134.47,130.28,1 28.89,128.63,128.56,122.90,118.38,116.69,116.51,111.82,100.96,65.52,65.46,64.75,54.93,48.63.HRMS(ESI):m / z calcd for C 26 H 22 ClN4O5 + [M+H] + :505.1274,found:505.1272

[0156] Compound 7-6-B: white solid, mp 191.4-192.0℃. 1 H NMR(500MHz,DMSO-d6)δ7.86–7.84(m,2H),7.62(d,J=7.5Hz,1H),7.53–7.47(m,5H ),7.42(d,J=1.9Hz,1H),7.35(d,J=12.5Hz,1H),6.98(s,1H),6.07(d,J=7.4Hz,1H) ,5.87(s,1H),4.93–4.83(m,1H),4.09(dd,J=14.0,3.0Hz,1H),4.03(td,J=11.7,5 .6Hz,2H),3.48–3.43(m,1H),3.24(d,J=10.7Hz,1H),3.02(td,J=13.1,4.2Hz,1H). 13C NMR (125MHz, DMSO-d6) δ184.59,170.83,170.55,162.43,162.18,151.66,139.56,137.88,134.13,132.23,131.27,131.22, 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

[0157] Example 8. Synthesis of 8-8

[0158]

[0159] Step 1: Synthesis of compound 8-2

[0160] Compound 8-1 (3.5 g, 24.5 mmol) was dissolved in a mixture of 10 mL of anhydrous DCM and 100 μL of anhydrous DMF, and oxalyl chloride (4.2 mL, 49 mmol) was slowly added dropwise, and the reaction solution was stirred at room temperature overnight. After the reaction was completed, the reaction solution was spin-dried to obtain a crude compound 8-2, which was directly used in the next step.

[0161] Step 2: Synthesis of compound 8-3

[0162] Dissolve compound 8-2 in 20 mL of anhydrous DCM, add dimethylhydroxylamine hydrochloride (3.0 g, 30 mmol), slowly add DIPEA (8.7 mL, 50 mmol) under stirring, and stir the reaction solution overnight at room temperature. After the reaction is completed, add 30 mL of 1M HCl to quench the reaction. Extract the aqueous phase with dichloromethane (2 × 50 mL), combine the organic phases and wash with saturated brine (1 × 50 mL), dry over anhydrous sodium sulfate, filter, and spin dry to obtain the crude product of 8-3. Finally, recrystallize from petroleum ether / ethyl acetate to obtain the pure product of compound 8-3 (3.72 g, yield 83%).

[0163] 1 H NMR (500MHz, CDCl3) δ8.77(s,1H),3.67(s,3H),3.34(s,3H),2.77(s,3H).

[0164] HRMS(ESI):m / z calcd for C7H11 N2O2S + [M+H] + :187.0536,found:187.0530

[0165] Step 3: Synthesis of compound 8-4

[0166] The magnesium ribbon with the surface oxide film removed in advance was cut into small pieces (1.46g, 60mmol) and added to 10mL of anhydrous THF. A few iodine particles were added under stirring, and the solution quickly turned brown-black. 10mL of THF solution of 2-bromo-5-fluorotoluene (1.26mL, 10mmol) was added to the reaction system, and the temperature was raised to reflux until the reaction was successfully initiated, and the solution color disappeared at this time, and it was close to colorless and transparent. After removing the heating, the reaction was stirred at room temperature, and the remaining 2-bromo-5-fluorotoluene (5mL, 40mmol) THF solution 10mL was continued to be added dropwise through a dropping funnel, and the system was kept slightly refluxed and continued to react for about 2h to obtain a 1M / L THF solution of 4-fluoro-2-methylphenylmagnesium bromide.

[0167] Compound 8-3 (2.8 g, 15 mmol) was dissolved in 10 mL of anhydrous THF, and 20 mL of the above-prepared 4-fluoro-2-methylphenylmagnesium bromide THF solution was added, and the reaction solution 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 continued to stir for 30 min to quench the reaction. The aqueous phase was extracted with ethyl acetate (2 × 50 mL), and the organic phases were combined and washed with saturated brine (1 × 50 mL), dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude product, which was purified by silica gel column chromatography to obtain pure compound 8-4 (1.65 g, yield 47%). 1 H NMR (500MHz, CDCl3) δ8.84 (s, 1H), 7.40 (dd, J=8.5, 5.8Hz, 1H), 7.02–6.93 (m, 2H), 2.54 (s, 3H), 2.39 (s, 3H). HRMS (ESI): m / z calcd forC 12 H 11 FNOS + [M+H] + :236.0540,found:236.0536

[0168] Step 4: Synthesis of compound 8-5

[0169] In an ice bath, compound 8-4 (0.8 g, 3.4 mmol) was added to 10 mL of methanol, and then sodium borohydride (0.26 g, 6.8 mmol) was slowly added under stirring, and the reaction solution was warmed to room temperature and continued to stir for 2 h. After the reaction was completed, 5 mL of water was added to quench the reaction, and then most of the methanol was removed by concentration under reduced pressure. The aqueous phase was extracted with dichloromethane (2 × 50 mL), and the organic phases were combined and washed with saturated brine (1 × 50 mL), dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain the crude product of 8-5. Finally, the pure product of compound 8-5 (0.8 g, yield 99%) was obtained by recrystallization from petroleum ether / ethyl acetate. 1 H NMR (500MHz, CDCl3) δ8.54 (s, 1H), 7.55 (dd, J=8.6, 5.9Hz, 1H), 6.94 (td, J=8.4, 2.7Hz, 1H), 6.85 (dd, J= 9.6,2.7Hz,1H),6.16(d,J=3.4Hz,1H),3.16(d,J=3.6Hz,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

[0170] Step 5: Synthesis of compound 8-6

[0171] Compound 8-5 (0.5 g, 2.1 mmol) was dissolved in 5 mL of anhydrous DCM, phosphorus tribromide (0.32 mL, 3.4 mmol) was slowly added, and the reaction was stirred at room temperature for 6 h. After the reaction was completed, a cold saturated sodium bicarbonate solution was added to the reaction system to quench the reaction, and the aqueous phase was extracted with dichloromethane (2×50 mL). The organic phases were combined and washed with saturated brine (1×50 mL), dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain the crude product 8-6, which was directly used in the next step. HRMS (ESI): m / z calcd for C 12 H 11 FNS + [M-Br] + :220.0591,found:220.0590

[0172] Step 6: Synthesis of compound 8-7

[0173] Compound 8-6 (0.3 g, 1 mmol) was added to 5 mL of anhydrous acetonitrile, and then (R)-7-benzyloxy-3,4,12,12A-tetrahydro-1H-[1,4]azino[3,4-C]pyrido[2,1-F][1,2,4]triazine-6,8-dione (0.27 g, 0.83 mmol) and CsF (0.32 g, 2.1 mmol) were added, and the reaction solution was heated to 70 ° C and stirred for 12 h. After the reaction was completed, the reaction solution was added to 25 mL of saturated sodium bicarbonate solution for quenching and extracted with ethyl acetate (2×50 mL). The organic phases were combined and washed with saturated brine (1×50 mL), dried over anhydrous sodium sulfate, filtered, and dried. The crude product was purified by column chromatography to obtain the racemic compound 8-7 (23 mg, yield 5%). HRMS (ESI): m / z calcd for C 29 H 28 N4O4FS + [M+H] + :547.1810,found:547.1804

[0174] Step 7: Synthesis of compound 8-8

[0175] Compound 8-7 (20 mg, 0.036 mmol) was added to 1 mL of N, N-dimethylacetamide, and lithium chloride (26 mg, 0.6 mmol) was added. The reaction solution 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 HCl aqueous solution 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 a crude product. Further recrystallization from isopropyl ether / chloroform gave a racemic compound 8-8 (11 mg, yield 65%).

[0176] White solid, mp 217.3-218.1 ° C. A mixture of diastereomers (1.7:1) was obtained in 65% yield. 1H NMR(500MHz,DMSO-d6)δ8.83(s,1.7H),8.11(s,1H),7.70(d,J=7.6Hz,1H),7.62(dd,J=6.9,3.1Hz,2H),7.56(dd,J=7.0,1.6Hz,1.7H),7.52(dd,J=8.6,6.2Hz,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=3.2Hz,1H),6.99–6.96(m,1.7H),6.23(d,J=7.6Hz,1H),6.07(d,J=7.5Hz,1.7H),6.06(s,1.7H),5.91(s,1H),5.07(d,J=2.8Hz,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=11.5,6.7,6.2,2.7Hz,5.1H),3.46(s,1H),3.44(d,J=2.6Hz,1.7H),3.25–3.22(m,1.7H),3.15(d,J=1.4Hz,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(125MHz,DMSO-d6)δ174.03,170.73,162.36,162.11(d,J=1.76Hz),160.17(d,J=2.65Hz),151.72,150.37,148.35,146.46,139.63,138.83(d,J=2.77Hz),137.73,137.46,137.40,136.18,135.9(d,J=2.4Hz),128.13,128.07,127.68,127.33,118.24,116.68,116.53,116.51(d,J=2.77Hz),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 forC 22 H 22N4O4FS + [M+H] + :457.1341,found:457.1336

[0177] Example 9. Synthesis of 9-8

[0178] Referring to the method of Example 8, except that 4-methylthiazole-5-carboxylic acid was replaced with 4-methyl-5-carboxylic acid-1,3-oxazole, the final product was a diastereoisomer mixture as shown in Formula 9-8, in which the ratio of the two diastereomers was 2:1 (yield 66%).

[0179] Compound 9-8: white solid, mp 229.6-230.3℃. 1 H NMR(500MHz,DMSO-d6)δ8.56(s,2H),8.10(s,1H),7.70(d,J=7.6Hz,1H),7.62(d d,J=6.9,2.3Hz,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=10.2,2.6Hz,2H),7.16(td,J=8.6,2.6Hz,2H),7.05( td,J=8.7,2.8Hz,1H),6.98(dd,J=10.0,2.8Hz,1H),6.23(d,J=7.6Hz,1H),6.07( d,J=7.5Hz,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=11.3,3.6,2.7Hz,6H),3.49–3. 46(m,2H),3.46(d,J=2.8Hz,1H),3.25–3.21(m,2H),3.17–3.13(m,1H),3.06–2.9 9(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(125MHz,DMSO-d6)δ184.10,174.03,170.75,164.16,162.36,160.20,155.95,153.78,150.38,148.75,146.47,14 5.82,144.37,139.96(d,J=8.8Hz),139.63(d,J=8.8Hz),137.67(d,J=7.3Hz),137.11(d,J=7.3Hz),135.93(d,J=3.2H z),133.90(d,J=2.8Hz),131.35,131.00,128.07,127.99,118.24,117.86,117.69,116.53,116.37,115.30,112.75,1 12.57,112.32,112.15,111.72,72.77,65.80,65.47,64.76,64.03,62.33,19.12,18.33,13.55,11.16.HRMS(ESI):m / z calcd for C 22 H 22 N4O5F + [M+H] + :441.1569,found:441.1572

[0180] Example 10. Synthesis of 10-7

[0181] Referring to the method of Example 8, except that 4-methylthiazole-5-carboxylic acid was replaced with 4-methylthiophene-5-carboxylic acid, the final product was a diastereoisomer mixture represented by Formula 10-7 (18 mg, yield 72%), in which the ratio of the two diastereomers was 1.6:1.

[0182] White solid, mp 209.2-210.0℃.. 1H NMR(500MHz,DMSO-d6)δ7.91(d,J=5.0Hz,1.6H),7.70(d,J=7.6Hz,1H),7.62(d,J=7.6Hz,1.6H),7.57(d,J=1.6Hz,1H),7.55(s,1H),7.49(dd,J=8.6,6.2Hz,1H),7.44(dd,J=8.5,5.9Hz,1.6H),7.38–7.35(m,2H),7.34–7.29(m,3.2H),7.26(d,J=5.1Hz,1H),7.21(dd,J=10.2,2.7Hz,1.6H),7.15–7.11(m,3.2H),7.01(td,J=8.7,2.9Hz,1H),6.97(dd,J=10.1,2.8Hz,1H),6.82(d,J=5.0Hz,1H),6.23(d,J=7.6Hz,1H),6.07(d,J=7.5Hz,1.6H),6.00(s,1.6H),5.75(s,1H),5.07(d,J=3.0Hz,2H),4.88(ddd,J=12.5,10.2,4.3Hz,1.6H),4.83–4.77(m,1H),4.17–4.07(m,3H),4.02(dq,J=11.3,3.4,2.7Hz,4.8H),3.44(s,2.6H),3.23(s,1H),3.16–3.12(m,1.6H),3.02(dd,J=4.2,1.5Hz,1H),2.95(ddd,J=13.7,12.1,4.3Hz,1.6H),2.28(s,4.8H),2.25(s,4.8H),2.17(s,3H),2.14(s,3H). 13C NMR (125MHz, DMSO-d6) δ189.57,174.03,170.71,163.65,162.36,161.68,155.94,151.61,148.74,145. 74,141.87,139.62,138.95,138.47,137.74,136.85,136.37,133.84,133.07,131.34,129.97,129.56,1 29.49,127.67,123.54,118.24,117.52,117.35,116.45,116.28,115.29,112.58,112.41,112.15,111. 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 / zcalcd for C 23 H 23 FN3O4S + [M+H] + :456.1388,found:456.1386

[0183] Example 11. Synthesis of 11-8-A and 11-8-B

[0184]

[0185] Step 1: Synthesis of compound 11-2

[0186] Pyridine (6.7 g, 84.6 mmol) was added to a DCM solution (100 mL) of compound 11-1 (10.0 g, 76.9 mmol), and then acetyl chloride (6.0 mL, 84.6 mmol) was added dropwise under stirring. After the addition was completed, the reaction solution was stirred at room temperature for 12 h. After the reaction was completed, the reaction solution was poured into 2M HCl (100 mL), extracted with DCM (3 × 100 mL), and the organic phases were combined. The organic phases were washed with water (1 × 100 mL), washed with saturated brine (1 × 100 mL), dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude product. Purification by silica gel column chromatography gave the pure compound 11-2 (12 g, yield 96%). 1 H NMR(500MHz, CDCl3)δ7.11–7.03(m,2H),6.95–6.88(m,1H),2.35(s,3H).HRMS(ESI):m / z calcdfor C8H6F2O2Na + [M+Na] +:195.0229,found:195.0228

[0187] Step 2: Synthesis of compound 11-3

[0188] At 0°C, compound 11-2 (12.0 g, 70 mmol) was dissolved in DCE (50 mL) solution, and then anhydrous aluminum chloride (9.3 g, 70 mmol) was added portionwise under stirring. After the addition was completed, the reaction solution was brought to reflux and stirred for 12 h. After the reaction was completed, 2M HCl (100 mL) was added to the reaction solution for quenching, and extracted with DCM (3×100 mL), and the organic phases were combined, and the organic phases were washed with water (1×100 mL), saturated brine (1×100 mL), dried over anhydrous sodium sulfate, filtered, and spin-dried. The obtained crude product was purified by silica gel column chromatography to obtain pure compound 11-3 (9.7 g, 81%). 1 H NMR (500MHz, CDCl3) δ12.57 (d, J=1.3Hz, 1H), 7.52 (ddd, J=9.2, 5.5, 2.3Hz, 1H), 6.72 (td, J=9.3, 6.6Hz, 1H), 2.62 (s, 3H). HRMS (ESI): m / z calcd for C8H6F2O2Na + [M+Na] + :195.0229,found:195.0231

[0189] Step 3: The synthesis of compounds 11-4 to 11-8 was carried out by referring to the method of Example 1, except that 4'-chloro-2'-hydroxyacetophenone (1-1) was replaced by compound 11-3, and finally two diastereoisomers corresponding to compound 11-8 were obtained: compound 11-8-A and compound 11-8-B.

[0190] Compound 11-8-A: white solid. 1H NMR(500MHz,DMSO-d6)δ11.68(s,1H),8.98(s,1H),7.20(t,J=7.6Hz,1H),7.15–7.08(m,1H),7 .06(d,J=7.7Hz,1H),5.99(s,1H),5.69(d,J=7.7Hz,1H),5.58(d,J=14.6Hz,1H),5.07(d,J=14. 7Hz,1H),4.81(dd,J=9.9,3.1Hz,1H),4.44(dd,J=13.5,2.5Hz,1H),3.78(dd,J=10.8,3.1Hz,1 H),3.73(dd,J=11.6,3.3Hz,1H),3.59(t,J=10.4Hz,1H),3.44–3.40(m,1H),3.16–3.10(m,1H). 13 C NMR(125MHz,DMSO-d6)δ177.04,160.96,157.33,156.81,151.65,138.22,127.02,126.90,124.06,123.93, 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

[0191] Example 12. Synthesis of 12-12-A and 12-12-B

[0192]

[0193] Step 1: Synthesis of compound 12-5

[0194] Compound 12-4 (4 g, 25.8 mmol) was dissolved in 15 mL of chlorobenzene, heated to 100 ° C under stirring, and then BPO (0.62 g, 2.58 mmol) and dibromohydantoin (8.87 g, 31.0 mmol) were added to the reaction solution, and the temperature was continued to be raised to reflux and reacted overnight. After the reaction was completed, the chlorobenzene was removed by vacuum concentration, the residue was dissolved in DCM, and the insoluble matter was removed by filtration. The filtrate was spin-dried to obtain a crude product, which was then purified by silica gel column chromatography to obtain a pure compound 12-5 (3 g, yield 51%). 1 H NMR (500MHz, CDCl3) δ8.59 (s, 1H), 4.56 (s, 2H), 4.48 (q, J = 7.1Hz, 2H), 1.45 (t, J = 7.1Hz, 3H). HRMS (ESI): m / zcalcd for C7H9NBrO3 + [M+H] + :233.9761,found:233.9759

[0195] Step 2: Synthesis of compound 12-6

[0196] Under nitrogen atmosphere, compound 12-5 (2 g, 8.6 mmol) and triphenylphosphine (2.7 g, 10.3 mmol) were added to 15 mL of toluene and 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 a crude compound 12-6, which was directly used 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

[0197] Step 3: Synthesis of compound 12-7

[0198] At 0°C, the crude compound 12-6 (1.5 g, 3.0 mmol) was dissolved in 10 mL of anhydrous acetonitrile, and then 2,3-difluorobenzaldehyde (0.85 g, 6.0 mmol) and DBU (0.55 g, 3.6 mmol) were added in sequence under stirring, and the reaction solution was heated to reflux and reacted overnight. After the reaction was completed, the solvent was removed by concentration under reduced pressure, and the pure compound 12-7 (0.8 g, yield 94%) was directly purified by silica gel column chromatography. 1H NMR(500MHz, CDCl3)δ8.09(d,J=0.9Hz,1H),7.11(dtd,J=10.0,7.7,2.0Hz,1H),7.06–6.93(m,2H),6.87 (d,J=12.0Hz,1H),6.72(d,J=12.0Hz,1H),4.45(q,J=7.1Hz,2H),1.43(t,J=7.2Hz,3H).HRMS(ESI):m / z calcd forC 14 H 12 NF2O3 + [M+H] + :280.0780,found:280.0773

[0199] Step 4: Synthesis of compound 12-8

[0200] Compound 12-7 (0.8 g, 2.9 mmol) was dissolved in ethanol / water (5 mL / 1 mL), and LiOH·H2O (0.24 g, 5.8 mmol) was added under stirring, and the mixture was reacted at room temperature for 2 h. After the reaction was completed, the ethanol was removed by concentration under reduced pressure, and 2M HCl was added dropwise to the residue under stirring until the pH of the system reached about 6. The mixture was filtered 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

[0201] Step 5: Synthesis of compound 12-9

[0202] Compound 12-8 (0.65 g, 2.6 mmol) was dissolved in 5 mL of anhydrous DCM, thionyl chloride (0.38 mL, 5.2 mmol) was added dropwise under stirring, and then a catalytic amount of DMF was added, and the reaction was refluxed 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 after the system returned to room temperature, and the temperature was raised to reflux and reacted overnight. After the reaction was completed, 10 mL of 1M HCl was added dropwise to the system to quench the reaction, and the aqueous phase was extracted with DCM (1×50 mL). The organic phases were combined and washed with saturated brine (1×50 mL), dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain the crude product of 12-9. Finally, the pure product of compound 12-9 (0.55 g, yield 92%) was obtained by silica gel column chromatography. 1H NMR (500MHz, CDCl3) δ8.97 (s, 1H), 8.35 (ddd, J = 9.2, 5.5, 2.0Hz, 1H), 7.37 (td, J = 8.9, 7.5Hz, 1H), 7.28 (s, 1H), 7.07 (d, J = 12.0Hz, 1H). HRMS (ESI): m / z calcd for C 12 H6NF2O2 + [M+H] + :234.0362,found:234.0363

[0203] Step 6: Synthesis of compound 12-10

[0204] In an ice bath, compound 12-9 (0.55 g, 2.36 mmol) was added to 10 mL of ethanol, and then sodium borohydride (0.09 g, 2.36 mmol) was slowly added under stirring, and the reaction solution was warmed to room temperature and continued to stir 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 concentration under reduced pressure. The aqueous phase was extracted with dichloromethane (2 × 50 mL), and the organic phases were combined and washed with saturated brine (1 × 50 mL), dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude product. Finally, the pure compound 12-10 (0.48 g, yield 87%) was obtained by recrystallization from ethyl acetate. 1 H NMR (500MHz, DMSO-d6) δ9.13 (s, 1H), 7.48 (dq, J=17.4, 8.5Hz, 2H), 7.04 (d, J= 11.8Hz,1H),6.97(d,J=11.8Hz,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

[0205] Step 7: Synthesis of compound 12-11

[0206] Compound 12-10 (0.4 g, 1.7 mmol) was added to a 50% (w / w)-T4P / EA (2 mL, 2.4 mmol) solution (n-butyl phosphoric acid (50% ethyl acetate solution)), and the mixture was stirred at room temperature. Then (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H-[1,4]azino[3,4-c]pyrido[2,1-f][1,2,4]triazine-6,8-dione (0.26 g, 0.8 mmol) and 2 mL of ethyl acetate were added, and the reaction solution was heated to 110°C and stirred for 5 hours. After the reaction was completed, the reaction solution was added to 15 mL of a saturated sodium bicarbonate solution for quenching and extracted with ethyl acetate (2×50 mL). After the organic phases were combined, they were washed with saturated brine (1×50 mL), dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The crude product was purified by column chromatography (dichloromethane: methanol = 100: 1 → 50: 1) to obtain 103 mg and 73 mg of pure compounds 12-11-A and 12-11-B, respectively (total yield 42%). The obtained compounds were detected by liquid chromatography column, and 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) with respect to *C.

[0207] Compound 12-11-A: 1 H NMR(500MHz,DMSO-d6)δ9.40(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),7.17–7.1 0(m,2H),7.07(dd,J=11.8,1.5Hz,1H),6.78(d,J=7.7Hz,1H),6.19(s,1H),5.72(d,J=7.7Hz,1H),5.25(d,J=11.0Hz,1H),5 .08(d,J=11.0Hz,1H),4.45(dd,J=13.5,2.5Hz,1H),4.22(dd,J=9.9,3.1Hz,1H),3.66(dd,J=11.6,3.3Hz,1H),3.53(dd,J= 10.7,3.1Hz,1H),3.29(d,J=10.3Hz,1H),3.22(td,J=11.7,2.7Hz,1H),2.97(ddd,J=13.4,11.7,3.5Hz,1H).HRMS(ESI):m / z calcd for C 29 H 23 N4F2O5 + [M+H]+ :545.1632,found:545.1631

[0208] Compound 12-11-B: 1 H NMR(500MHz,DMSO-d6)δ9.26(s,1H),7.66(dd,J=8.8,5.1Hz,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=11.9Hz,1H),7.04(dd,J=11.8,1.5Hz,1H),6.74(d,J=7.6Hz,1H),6.13(s,1H),5.84(d,J=7.7 Hz,1H),5.17(d,J=10.8Hz,1H),5.10(d,J=10.8Hz,1H),4.39(dd,J=13.4,2.5Hz,1H),4.09(dd,J=9.8,3.1Hz,1H),3.62(dd ,J=11.6,3.3Hz,1H),3.55(dd,J=10.6,3.1Hz,1H),3.22–3.12(m,2H),2.86(ddd,J=13.5,11.7,3.5Hz,1H).HRMS(ESI):m / z calcd forC 29 H 23 N4F2O5 + [M+H] + :545.1632,found:545.1628

[0209] Step 8: Synthesis of compound 12-12-A

[0210] 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 solution 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 HCl aqueous solution 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 a crude compound 12-12-A. Further recrystallization from isopropyl ether / chloroform gave a pure compound 12-12-A (70 mg, yield 90%).

[0211] White solid. 1H NMR (500MHz, DMSO-d6) δ11.70(s,1H),9.41(s,1H),7.38–7.32(m,1H),7.28(dd,J=9. 0,5.1Hz,1H),7.17–7.06(m,2H),6.69(d,J=7.7Hz,1H),6.21(s,1H),5.56(d,J=7.6H z,1H),4.43(dd,J=13.3,2.4Hz,1H),4.24(dd,J=9.4,3.7Hz,1H),3.68(dd,J=11.6,3 .4Hz,1H),3.56–3.48(m,2H),3.40–3.37(m,1H),3.05(ddd,J=13.3,11.7,3.5Hz,1H). 13 C NMR(125MHz,DMSO-d6)δ171.60,161.54,160.99,158.79,153.38,138.92,129.12,128.93,126.50,120.71,1 20.53,120.44,117.69,117.56,116.13,115.54,110.93,70.12,68.23,65.96,63.56,45.82.HRMS(ESI):m / z calcd for C 22 H 17 N4F2O5 + [M+H] + :455.1162,found:455.1160

[0212] Step 6: Synthesis of compound 12-12-B

[0213] 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 solution 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 HCl aqueous solution and 1 mL of water were added, and stirring was continued for 1 h. The solid precipitated from the reaction solution was collected by suction filtration and dried to obtain a crude compound 12-12-B. Further recrystallization by isopropyl ether / chloroform gave a pure compound 12-12-B (50 mg, yield 94%). White solid. 1H NMR (500MHz, DMSO-d6) δ11.68(s,1H),9.27(s,1H),7.70(dd,J=8.9,5.1Hz,1H),7.60(q,J=8 .8Hz,1H),7.12(d,J=11.8Hz,1H),7.06(dd,J=11.7,1.5Hz,1H),6.66(d,J=7.6Hz,1H),6.16 (s,1H),5.69(d,J=7.6Hz,1H),4.45–4.33(m,1H),4.14(dd,J=9.7,3.2Hz,1H),3.64(dd,J=1 1.7,3.2Hz,1H),3.55(dd,J=10.7,3.2Hz,1H),3.47(m,1H),3.35(m,1H),2.99–2.90(m,1H). 13 C NMR(125MHz,DMSO-d6)δ171.33,160.97,159.79,157.64,153.10,138.21,128.76,128.53,125.68,125.61,1 20.30,119.88,117.89,117.75,115.53,115.46,110.70,69.18,67.73,65.43,63.43,45.35.HRMS(ESI):m / z calcd for C 22 H 17 N4F2O5 + [M+H] + :455.1162,found:455.1166.

[0214] Example 13. Synthesis of 13-16-A and 13-16-B

[0215]

[0216] Step 1: Synthesis of compound 13-10

[0217] The synthesis of compound 13-10 was carried out according to the method in Example 12.

[0218] Step 2: Synthesis of compound 13-11

[0219] At 0°C and nitrogen atmosphere, compound 13-5 (1,2-bis(2,3-difluorophenyl) disulfide) (0.6 g, 2 mmol) was dissolved in acetonitrile / water (8 mL / 2 mL), and zinc powder (0.52 g, 8 mmol) was added. Under stirring, ferric chloride (0.65 g, 4 mmol) was added in batches, the reaction solution was restored to room temperature and the reaction was continued for 3 h. Then compound 13-10 (0.9 g, 4 mmol) was added and reacted overnight. After the reaction was completed, the insoluble matter was filtered off through diatomaceous earth, and the filter cake was washed with EA. The filtrate was collected and washed with water (1×50 mL) and saturated brine (1×50 mL) in turn. After drying over anhydrous sodium sulfate and concentrating under reduced pressure, the crude product was obtained, and then purified by silica gel column chromatography (petroleum ether: ethyl acetate = 50:1) to obtain pure compound 13-11 (0.7 g, yield 91%).

[0220] 1 H NMR (500MHz, CDCl3) δ8.28 (s, 1H), 7.13–6.97 (m, 3H), 4.46 (q, J = 7.1Hz, 2H), 4.19 (d, J = 0.9Hz, 2H), 1.43 (t, J = 7.2Hz, 3H).

[0221] HRMS(ESI):m / z calcd for C 13 H 12 SF2NO3 + [M+H] + :300.0501,found:300.0495

[0222] Step 3: Synthesis of compound 13-12

[0223] 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, and the mixture was reacted at room temperature for 2 h. After the reaction was completed, the ethanol was removed by concentration under reduced pressure, and 2M HCl was added dropwise to the residue under stirring until the pH of the system reached about 6. The mixture was filtered and the filtrate was discarded. The filter cake was washed with water and dried to obtain 0.56 g of crude compound 13-12, which was directly used in the next step without further purification.

[0224] HRMS(ESI):m / z calcd for C 11 H8SF2NO3 + [M+H] + :272.0188,found:272.0184

[0225] Step 4: Synthesis of compound 13-13

[0226] 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, the reaction solution was slowly dripped into a cold saturated sodium bicarbonate solution, and the aqueous phase was extracted with EA (2×100 mL). The organic phases were combined and washed with saturated brine (1×50 mL). After drying over anhydrous sodium sulfate, the crude product was spin-dried to obtain a crude product. After purification by silica gel column chromatography, the pure product of compound 13-13 (0.2 g, yield 46%) was obtained.

[0227] 1 H NMR (500MHz, CDCl3) δ8.49 (s, 1H), 7.88 (ddd, J = 8.9, 5.3, 1.9Hz, 1H), 7.30–7.27 (m, 1H), 3.95 (s, 2H).

[0228] HRMS(ESI):m / z calcd for C 11 H6SF2NO2 + [M+H] + :254.0082,found:254.0082

[0229] Step 5: Synthesis of Compound 13-14

[0230] In an ice bath, compound 13-13 (0.2 g, 0.8 mmol) was added to 10 mL of ethanol, and then sodium borohydride (0.06 g, 1.6 mmol) was slowly added under stirring, and the reaction solution was warmed to room temperature and continued to stir and react 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 concentration under reduced pressure. The aqueous phase was extracted with dichloromethane (2 × 50 mL), and the organic phases were combined and washed with saturated brine (1 × 50 mL), dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude product. Finally, the pure compound 13-14 (0.18 g, yield 90%) was obtained by recrystallization from ethyl acetate.

[0231] 1 H NMR (500MHz, DMSO-d6) δ8.69(s,1H),7.54–7.42(m,2H),6.62(d,J=4.6Hz,1H),6.33(d,J=4.6Hz,1H),4.08–3.98(m,2H).

[0232] HRMS(ESI):m / z calcd for C 11 H6SF2NO + [M-OH] + :238.0133,found:238.0131

[0233] Step 6: Synthesis of compounds 13-15

[0234] Compound 13-14 (0.18 g, 0.7 mmol) was added to a 50% (w / w)-T4P / EA (2 mL, 2.4 mmol) solution (n-butyl phosphoric acid (50% ethyl acetate solution)), and then (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H-[1,4]azino[3,4-c]pyrido[2,1-f][1,2,4]triazine-6,8-dione (0.16 g, 0.5 mmol) and 2 mL of ethyl acetate were added, and the reaction solution was heated to 110° C. and stirred for 5 hours. After the reaction was completed, the reaction solution was added to 15 mL of a saturated sodium bicarbonate solution for quenching and extracted with ethyl acetate (2×50 mL). After the organic phases were combined, they were washed with saturated brine (1×50 mL), dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The crude product was purified by column chromatography to obtain 20 mg and 14 mg of pure compounds 13-15-A and 13-15-B, respectively (total yield 12%). The obtained compounds were detected by liquid chromatography column, and the retention time of compound 13-15-A was 7.02 min, and the retention time of compound 13-15-B was 7.04 min. Compound 13-15-A and compound 13-15-B are two non-corresponding isomers of the above formula (13-15) with respect to *C.

[0235] Compound 13-15-A: HRMS (ESI): m / z calcd for C 28 H 22 N4F2O5S + [M+H] + :565.1352,found:565.1350

[0236] Compound 13-15-B: HRMS (ESI): m / z calcd for C 28 H 22 N4F2O5S + [M+H] + :565.1352,found:565.1349

[0237] Step 8: Synthesis of Compound 13-16-A

[0238] 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 solution 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 HCl aqueous solution 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 a crude compound 13-16-A. Further recrystallization from isopropyl ether / chloroform gave a pure compound 13-16-A (15 mg, yield 90%).

[0239] White solid.HRMS(ESI):m / z calcd for C 21 H 17 N4F2O5S + [M+H] + :475.0883,found:475.0880

[0240] Step 9: Synthesis of compound 13-16-B

[0241] 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 solution 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 HCl aqueous solution 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 a crude compound 13-16-B. Further recrystallization from isopropyl ether / chloroform gave a pure compound 13-16-B (11 mg, yield 90%).

[0242] White solid.HRMS(ESI):m / z calcd for C 21 H 17 N4F2O5S + [M+H] + :475.0883,found:475.0878

[0243] Example 14. Synthesis of 14-15-A and 14-15-B

[0244] Referring to the method of Example 13, two diastereomers corresponding to compound 14-15 were finally obtained: compound 14-15-A and compound 14-15-B (see the claims or description for the structure, the same below).

[0245] Compound 14-15-A: HRMS (ESI): m / z calcd for C 21 H 17 N4F2O5Se + [M+H] + :523.0327,found:523.0325

[0246] Compound 14-15-B: HRMS (ESI): m / z calcd for C 21 H 17 N4F2O5Se + [M+H] + :523.0327,found:523.0330

[0247] Example 15. Synthesis of 15-12-A and 15-12-B

[0248] By referring to the method of Example 12, two diastereomers corresponding to compound 15-12 were finally obtained: compound 15-12-A and compound 15-12-B.

[0249] Compound 15-12-A: HRMS (ESI): m / z calcd for C 22 H 19 N4O5[M+H] + :419.1350,found:419.1352

[0250] Compound 15-12-B: HRMS (ESI): m / z calcd for C 22 H 19 N4O5[M+H] + :419.1350,found:419.1353

[0251] Example 16. Synthesis of 16-12-A and 16-12-B

[0252] By referring to the method of Example 12, two diastereomers corresponding to compound 16-12 were finally obtained: compound 16-12-A and compound 16-12-B.

[0253] Compound 16-12-A: HRMS (ESI): m / z calcd for C 22 H 18 FN4O5[M+H] + :437.1256,found:437.1255

[0254] Compound 16-12-B: HRMS (ESI): m / z calcd for C 22 H 18 FN4O5[M+H] + :437.1256,found:437.1257

[0255] Example 17 Inhibitory effect of the compounds disclosed in the present invention on influenza virus polymerase expressed in HEK-293T cells

[0256] HEK-293T cells were trypsinized and counted, then suspended in DMEM medium containing 10% fetal bovine serum and then cultured at 1.5 × 10 4 cells / well were inoculated in a 96-well plate, with 100 μL per empty volume. The 96-well plate was cultured at 37°C / 5% CO2 for 24 hours. After 24 hours, 20 μL of DMEM solution containing a certain concentration of the compound to be tested was added, and 20 μL of DMEM solution was added to the control well, and the culture was continued in the cell culture incubator for 2 hours. Subsequently, a plasmid transfection solution (plasmids including PA, PB1, PB2, NP, Luciferase and Renilla) was added to each well, and the culture was continued for 24 hours. The culture medium was removed, and 100 μL / well of PBS solution was added to gently wash the cells. After removing the PBS, 1×CLB cell lysate was added to each well. After shaking at room temperature for 20 minutes, the lysate in the plate was transferred to a 1.5 mL centrifuge tube, centrifuged at 12000 rpm for 2 minutes, 20 μL of the supernatant was taken and placed in a 96-well white plate, and the Luciferase substrate and Renilla substrate were added in turn, and the luminescence value was detected using an enzyme reader. The regression curve and regression equation were obtained by regression analysis, and the IC of the compound described in the present invention was calculated. 50 In Table 1, PA / WT represents wild-type influenza PA; PA / I38T represents I38T mutant influenza PA.

[0257] Table 1. IC values ​​of the hydroxypyridone compounds and baloxavir of the present invention 50 Value and RI value

[0258]

[0259]

[0260] Examples 1 to 16 and IC of baloxavir 50The values ​​and RI values ​​are shown in Table 1. The results show that the hydroxypyridone compounds of the present invention can effectively inhibit influenza A virus RNA polymerase in HEK-293T cells, especially the compounds prepared in Example 14 and Example 15, but the inhibitory activity of baloxavir on PA / I38T resistant strains is significantly reduced. The hydroxypyridone compounds disclosed in the present invention have a resistance index RI lower than that of baloxavir, indicating that the derivatives of the present invention are less affected by influenza A PA / I38T mutant strains and are superior to baloxavir.

[0261] Example 18 Antiviral effect of the compounds disclosed in the present invention in MDCK cells infected with influenza A virus

[0262] MDCK cells were trypsinized and counted, then suspended in DMEM medium containing 10% fetal bovine serum and then cultured at 1.5×10 5 MDCK cells were inoculated in 24-well plates, with a medium volume of 500 μL per well. The 24-well plates were cultured at 37°C / 5% CO2 for 24 hours, and the medium was removed and 500 μL of new medium was added. After adding a certain concentration of the test compound, the influenza virus was inoculated in the 24-well plates at a multiplicity of infection (MOI = virus number / cell number) of 0.1, and the plates were cultured in a cell culture incubator for 24 hours. The supernatant was collected and the virus TCID was determined. 50 The regression curve and regression equation are obtained by regression analysis, and the EC value of the compound of the present invention is calculated. 50 value.

[0263] Table 2. EC values ​​of the hydroxypyridone compounds and baloxavir of the present invention 50 value

[0264]

[0265] Examples 1, 2, 12 and EC of baloxavir 50 The values ​​are shown in Table 2. The results show that the hydroxypyridone compounds (1-6-A, 1-6-B, 2-6-A, 2-6-B, 12-12-A, 12-12-B) obtained in the present invention can effectively inhibit influenza A virus in MDCK cells, and Examples 1, 2, and 12 have an inhibitory activity comparable to that of baloxavir against wild-type strains, and Example 12 has an inhibitory activity significantly superior to that of baloxavir against I38T mutant strains.

Claims

1. A pyridone derivative, characterized in that: Having the structure shown by formula (I), formula (II) and formula (III), or a pharmaceutically acceptable salt thereof, in R1 is selected from hydrogen, halogen, C1-C6 alkyl, R1 is one or multiple independent of each other; R2 is selected from non-existent, C1-C6 alkyl or C1-C3 alkyl acyl or R3 is 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 does not exist; Ring A is selected from one of the following structures:

2. The pyridone derivative according to claim 1, characterized in that In formula (I), formula (II) and formula (III), the C atom corresponding to * is in S configuration or R configuration or a mixture of S configuration and R configuration: R3, R4 and X are as defined in claim 1; R1 is H, halogen, C1-C3 alkyl, R1 is 1 or 2 independent of each other; R2 is selected from the group consisting of not existing, C1-C3 alkyl, 3. The pyridone derivative according to claim 1 or 2, characterized in that It has the structure represented by the following formula (Ia), (Ib), (Ic), (III-a): R1 is H, halogen, C1-C3 alkyl, R1 is 1 or 2 independent of each other, R3 is H or 4. The pyridone derivative according to claim 1, characterized in that Having any of the following structures:

5. An intermediate for preparing the pyridone derivative according to any one of claims 1 to 4, characterized in that: Has a structure shown in any of the following formulas: R1, R2, R4 and X are as defined in claim 1.

6. The intermediate according to claim 5, characterized in that Has a structure shown in any of the following formulas:

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

8. The method for synthesizing a pyridone derivative according to claim 7, characterized in that: Ring A is When the intermediate shown in formula (II-2) first reacts with PBr3 to obtain intermediate (II-2-a): Then, the intermediate (II-2-a) reacts with (R)-7-benzyloxy-3,4,12,12a-tetrahydro-1H-[1,4]azino[3,4-c]pyrido[2,1-f][1,2,4]triazine-6,8-dione in the presence of CsF to obtain the intermediate represented by formula (II-3).

9. The method for synthesizing a pyridone derivative according to claim 7, characterized in that: When the intermediate (I-1) is a structure represented by formula (I-1-1), the intermediate (I-1) is obtained by reacting the intermediate (I-1-1-1) with tert-butyl nitrite: When the intermediate (I-1) is a structure represented by formula (I-1-2), the intermediate (I-1) is obtained by reacting the intermediate (I-1-2-1) with tert-butyl nitrite: When the intermediate (I-1) is a structure represented by formula (I-1-3), the intermediate (I-1) is obtained by reacting the intermediate (I-1-3-1) with polyphosphoric acid: When the intermediate (II-1) is a structure represented by formula (II-1-1), the intermediate (II-1) is obtained by reacting the intermediate (II-1-1-1), phenylacetylene and tert-butyl nitrite: When the intermediate (II-1) is a structure represented by formula (II-1-2), the intermediate (II-1) is obtained by reacting the intermediate (II-1-2-1) with a Grignard reagent corresponding to formula (II-1-2-2) under the action of I: When the intermediate (III-1) is a structure represented by formula (III-1-1), the intermediate (III-1) is obtained by first reacting the intermediate (III-1-1-1) with thionyl chloride and then reacting with aluminum chloride: Wherein, Z1 is S or O; Z2 is CH or N, X1 is S or Se; R2 is selected from 10. Use of the pyridone derivative according to any one of claims 1 to 4 in the preparation of influenza drugs.

Images